PMID- 19804190 TI - Peripheral arterial disease training: "Vascular medicine for the interventionalist". PMID- 19804191 TI - Future directions in antiarrhythmic drug therapy for atrial fibrillation. AB - Atrial fibrillation is the most commonly sustained cardiac arrhythmia. Drugs currently approved by the US FDA for the treatment of this arrhythmia are imperfect owing to either side effects or limited efficacy. Drug development strategies have focused on two areas: the modification of existing agents--such as Class III drugs aimed at improving their safety and efficacy profile--and targeting newly postulated mechanisms of atrial fibrillation. In this article, we review new drugs currently in development and promising drug strategies for atrial fibrillation prevention and treatment. PMID- 19804192 TI - Future strategies in the management of coronary artery disease. AB - New diagnostic techniques have now become available that will enable us to obtain information on coronary artery disease (CAD), which was not previously possible. This is urgently needed, since changes in CAD management are highly desirable considering that up to 60% of patients die due to an insufficient diagnostic strategy (reactive strategy). In this article, the pathophysiology of CAD and the evidence of current concepts on coronary atherosclerosis are critically reviewed. Consequently, a shift from a 'reactive strategy' of CAD management to an active strategy is proposed in order to detect and treat patients before acute myocardial infarcts occur. Reliability, repeatability, cost-effectiveness and comfort are important features of an optimal test for this active strategy. This perspective gives an overview of current and future tests with respect to these features. The novel tomographic techniques are particularly promising with respect to standardization and reproducibility, which will set the basis for the determination of test performance and, consequently, cost-effectiveness. Assessment of cost-effectiveness is increasingly important, since prevalence of CAD is high and will most likely increase, while resources will remain restricted in most countries. Considering these economic restrictions, this article also proposes new mathematical tools using data from prospective trials and large international registries as an objective means to select the best tests for CAD diagnosis and management, and to identify those patient subsets which benefit most from an active strategy. PMID- 19804193 TI - Will carotid stenting eclipse carotid endarterectomy and, if so, when and how? AB - Treatment of carotid artery stenosis to prevent stroke ranks amongst the most significant roles for vascular surgeons and specialists as a whole. Carotid endarterectomy (CEA) is the acknowledged standard of care, and is based on solid scientific evidence. However, its pedestal position has been challenged on several fronts, mainly because approximately 20% of such operations are performed on patients who may be at high-risk for poor outcomes (or higher than usual complications) owing to unfavorable factors related to anatomical and/or medical comorbidity factors. Additionally, and as attitudes towards less invasion reach the carotid territory, CEA is increasingly viewed with some suspicion (and even disdain) by those who glorify everything endovascular simply for being so. The result is that percutaneous carotid-stent intervention has evolved into a formidable competitor to CEA. But it will not have a free ride. While it is arguably true that the high-risk territory has been conquered by carotid angioplasty stenting (CAS), the bulk of the CEA market remains unaffected. Treatment of nonhigh-risk patients with CAS remains highly controversial and essentially unproven, especially for those who are asymptomatic. CEA remains the only intervention whose application is based on level 1 evidence. Thus, it would be fair to say that rise of CAS to domination has a long way to go. PMID- 19804194 TI - Stress--the battle for hearts and minds: links between depression, stress and ischemic heart disease. AB - Depression and ischemic heart disease (IHD) are strongly related common disorders. Depression itself is an independent cardiac risk factor and is associated with a two- to threefold increase in IHD mortality. Attention has now shifted to identifying the common underlying mechanisms that could make individuals susceptible to both disorders. Abnormalities that have been implicated in this relationship include abnormal platelet activation, decreased baroreceptor sensitivity and endothelial dysfunction. Depression and IHD both have a high association with environmental stress, and depression is characterized by abnormalities of the stress-hormone axis. This review provides a brief overview of some recent developments in our understanding of the pathophysiological links between stress, depression and IHD. PMID- 19804195 TI - Genetics of atrial fibrillation. AB - Atrial fibrillation (AF) is the most common arrhythmia encountered in clinical practice. Despite its prevalence, relatively little is known regarding the primary mechanisms of AF and, therefore, current treatment practices focus mainly on controlling the disorder and preventing its complications once it is already present. The study of the pathogenesis of AF is complicated by the varied clinical presentation of the arrhythmia and its coexistence with other cardiac pathologies. This article reviews current efforts to delineate the fundamental mechanisms of AF, with a focus on genetic studies. Identification of the underlying etiology may result in the development of more targeted and effective therapies for AF. PMID- 19804196 TI - Vascular stem cells: a new concept in the pathogenesis of atherosclerosis and interventions for coronary heart disease. AB - Vascular stem cells are undifferentiated, oligopotent progenitor cells that are capable of giving rise to mature, functional cells in the vascular wall. Several types of vascular progenitor cells have been identified and characterized from embryonic and adult tissues, including progenitors with the potential to differentiate into endothelial and smooth muscle cells. The progenitors for endothelial and smooth muscle cells reside in atherosclerotic or restenotic lesions and circulate in the bloodstream. These stem cells may malfunction under the influence of the risk factors for atherosclerosis, as well as by medical interventions. The biological activities of these stem cells contribute to the regeneration, repair and remodeling of arterial walls injured by atherosclerosis. Hypercholesterolemia, inflammation, mechanical stress and genetic defects may interact in regulating the vascular stem cell response to atherogenic stimulation. Stem cell production, potency, growth and differentiation may decline as people age. Clarifying the cellular and molecular pathways that govern stem cell growth, differentiation and apoptosis should help clinical scientists to understand the pathogenesis of atherosclerosis and to develop novel therapeutic strategies for coronary heart disease. Recent clinical trials demonstrate encouraging outcomes of stem cell therapies. PMID- 19804197 TI - Stroke in the fetus and neonate. AB - Multiple patterns of focal ischemic injury in the fetal and perinatal brain are now recognized as important causes of neurological disability in children. Strokes may be arterial or venous, symptomatic or subclinical, and can occur across multiple timeframes. Establishing the importance of an increasing number of potential risk factors and translating that knowledge into an improved understanding of pathophysiology are major challenges. Advanced neuroimaging has facilitated increased recognition of perinatal stroke and an improved understanding of plasticity mechanisms in the developing brain. Better outcome prediction will assist in patient selection for advancing neuroprotective strategies to reduce cerebral palsy and other adverse outcomes suffered by most children. PMID- 19804199 TI - Heart size matters: lessons with MRI. PMID- 19804198 TI - Elucidation of an SRE-1/SREBP-independent cellular pathway for LDL-receptor regulation: from the cell surface to the nucleus. AB - Reduction in blood levels of low-density lipoprotein (LDL) cholesterol lowers the risk of coronary heart disease. The elucidation of cellular pathways that control LDL-receptor expression through a cholesterol-mediated negative feedback mechanism has provided a crucial molecular basis for the development and clinical applications of statins in the treatment of hypercholesterolemia. The characterization of signaling transduction pathways elicited by cytokine oncostatin M (OM) in liver cells has revealed a novel cellular pathway that activates LDL-receptor transcription independent of intracellular levels of cholesterol and sterol-regulatory element binding proteins. This transcriptional activation is achieved through interactions of the sterol-independent regulatory element of LDL-receptor promoter and transcription factors Egr1 and c/EBPbeta, and is dependent upon the activation of the extracellular signal-regulated kinase signaling cascade by OM. In vivo OM administration in hyperlipidemic animals reduces circulating cholesterol and prevents lipid accumulation in the liver. Exploring this sterol-independent cellular pathway may lead to new therapeutic advances. PMID- 19804200 TI - S100A1: a calcium-modulating inotropic prototype for future clinical heart failure therapy. PMID- 19804202 TI - Gadofosveset: the first intravascular contrast agent EU-approved for use with magnetic resonance angiography. AB - Gadofosveset is the first intravascular contrast agent approved for use with magnetic resonance angiography in the European Union. Gadofosveset reversibly binds to albumin, providing extended intravascular enhancement beyond that of existing extracellular magnetic resonance contrast agents. Prior to approval, gadofosveset underwent extensive testing to evaluate the safety and efficacy of the drug; the clinical trials program included blinded, placebo-controlled, dose ranging, efficacy in a variety of vascular beds (areas of turbulent blood flow, renal and pedal), examination of potential drug interaction with warfarin and comparison with x-ray angiography. The clinical trials demonstrate that gadofosveset-enhanced magentic resonance angiography is: safe and well tolerated in patients with vascular disease; effective for the detection of vascular stenosis and aneurysms; significantly more accurate (both more sensitive and specific) than noncontrast magnetic resonance angiography for the diagnosis of vascular stenoses; and similar to conventional angiography for the overall characterization of vascular disease, without the need for catheterization. PMID- 19804203 TI - CHEST 2006 Overview. PMID- 19804204 TI - Highlights of the late-breaking clinical trials from the 2006 Scientific Sessions of the American Heart Association. PMID- 19804205 TI - Pulmonary valve replacement: extending the indications to the whole spectrum of pulmonary valve disease. AB - Pulmonary valve replacement through a transcatheter technique is currently limited to the small group of patients with a right ventricular outflow tract that does not exceed 22-26 mm in diameter. In this report, we will give an overview of ongoing work to extend the present indication to the vast majority of patients. Second-generation devices and approaches are described, both of which prepare patients to receive the currently available valved stents. The hybrid approach was the first to be applied to humans, but remains highly invasive and justifies research into pure transcatheter strategy. Therefore, it is clear that the transcatheter approach will soon represent the only valuable alternative to on-pump pulmonary valve replacement. PMID- 19804206 TI - Remote endarterectomy for occlusive iliac and superficial femoral artery disease. AB - Today's patients increasingly desire a low-risk procedure with rapid return to functional status following surgery. Many patients actively seek a minimally invasive option. During the last decade, minimally invasive surgical and radiological procedures emerged from their infancy to become an integral part of the treatment strategy in both peripheral arterial occlusive disease and aneurysmal arterial disease. The trend towards restoration of luminal patency, using the vessel itself as a conduit, can also be termed restorative intervention. As radiological percutaneous transluminal angioplasty has thus far proven unsuccessful in the long term for iliac and femoropopliteal occlusive lesions exceeding 10 cm, a new endovascular surgical procedure has been developed. A modification to the original ringstripper, used in semiclosed endarterectomy, was made to enable the technique of remote endarterectomy. PMID- 19804207 TI - Reactive oxygen species, cell growth, cell cycle progression and vascular remodeling in hypertension. AB - Reactive oxygen species (ROS) include superoxide, hygrogen peroxide and hydroxyl radical. Under physiological conditions, all vascular cell types produce ROS in a controlled and regulated fashion, mainly through nonphagocyte NADPH oxidase. An imbalance between pro-oxidants and antioxidants results in oxidative stress. ROS are important intracellular signaling molecules. There is growing evidence that increased oxidative stress and associated oxidative damage are mediators of vascular injury in hypertension, as well as in other cardiovascular diseases. Oxidative stress causes vascular injury by reducing nitric oxide bioavailability, altering endothelial function and vascular contraction/dilation, promoting vascular smooth muscle cell proliferation and hypertrophy, and increasing extracellular matrix deposition and inflammation. The present review focuses on the regulatory role of ROS on cell growth and cell cycle progression and discusses implications of these events in vascular remodeling in hypertension. PMID- 19804208 TI - Current trends in the management of small abdominal aortic aneurysms. AB - Over the past decade, there have been revolutionary changes in the diagnosis and treatment of abdominal aortic aneurysms. Minimally invasive endovascular technologies have decreased length of stay and, more importantly, improved at least the early outcomes compared with conventional open surgical therapy. Several large, randomized, prospective trials have re-evaluated the optimal size to treat abdominal aortic aneurysms. The guidelines and conclusions from these studies, the UK Small Aneurysm Trial and The Veterans Affairs Administration aneurysm Detection And Management Trial recommend intervening on aortic aneurysms when they reach 5.5 cm in transverse diameter. However, these studies do not bear direct relevance to contemporary management of aneurysms with endovascular techniques. Clinical trials are currently underway to assess the optimal size for treating abdominal aortic aneurysms with endovascular stent grafts. PMID- 19804209 TI - Blood flow devices in medical research and clinical testing in humans: are we approaching personalized medicine? AB - This review focuses on studies of blood flow devices employed in man to unravel the mechanisms of bleeding and thrombotic disorders, and on the characterization of novel experimental antithrombotic entities and drug candidates in biopharmaceutical research and development. Clinical studies with drug candidates and new therapeutic strategies have also been performed, and the predictability of these experimental approaches to clinical situations is excellent. Based on the solid validation of these flow devices, miniature flow devices employing nonanticoagulated blood drawn directly from an antecubital vein should be developed for diagnostic purposes. It is anticipated that such a diagnostic flow device could develop into a personalized medicine approach. PMID- 19804210 TI - New approach to prevent myocardial hypertrophy: the import blocking peptide. AB - Calcineurin, a serine/threonine phosphatase, plays a crucial role in the development of myocardial hypertrophy. Calcineurin is a cytosolic phosphatase that dephosphorylates the nuclear factor of activated T cells (NFAT), a transcription factor. Until now, it has been postulated that dephosphorylated NFAT is shuttled into the nucleus. Recent evidence demonstrates that not only NFAT, but also calcineurin, is localized in the nucleus. Once calcineurin and NFAT enter the nucleus of cardiomyocytes, transcription of genes that are characteristic for myocardial hypertrophy (e.g., brain natriuretic peptide and atrial natriuretic peptide) occurs. Although the exact nuclear function of calcineurin remains unclear, its co-existence with NFAT is important for the full transcriptional activity of the calcineurin/NFAT signaling cascade. The principal effect of nuclear calcineurin is likely the prolonged nuclear retention period of NFAT. Potential effects of nuclear calcineurin include an antagonistic function to glycogen synthase kinase 3beta, which phosphorylates NFAT for its export out of the nucleus, or direct antagonization of the export of NFAT, catalyzed by the chromosome region maintenance 1, which would leave NFAT nuclear. The nuclear localization sequence (NLS) region at the amino acid sequence from position 172 to 183 of calcineurin Abeta is essential for shuttling calcineurin into the nucleus by importinbeta(1). A synthetic import blocking peptide (IBP) that mimics the nuclear localization sequence of calcineurin was generated. The NLS analog on IBP saturates the calcineurin binding site of importinbeta(1). This prevents the binding of calcineurin to importin and inhibits the nuclear shuttling of calcineurin. Inhibition of the calcineurin/importinbeta(1) interaction by competing synthetic peptides represents a new approach to the inhibition of the development of myocardial hypertrophy. PMID- 19804211 TI - Echocardiographic detection of dysfunctions in donor hearts. AB - Heart transplantation activity is steadily declining worldwide, despite initiatives to increase the donor pool. The yield of transplantable hearts remains low, leading to wastage of over two-thirds of donor organs. Accurate assessment and intervention is required to increase the number of transplantable hearts. The retrieval rate could be increased if techniques become available to differentiate between transplantable and nontransplantable hearts. Echocardiography is an ideally suited noninvasive investigation that may guide donor optimization. However, there are technical difficulties in acquiring high quality views, and changes in ventricular function following catecholamine storm in brainstem death often lead to misinterpretation of standard 2D images. Current advances in technology, including the routine use of left ventricular contrast, 3D imaging and tissue doppler studies, may improve the accuracy, and thereby the utilization, of echocardiography in donor heart assessment. PMID- 19804213 TI - Percutaneous coronary interventions: benefit, risk and cost-effectiveness. PMID- 19804212 TI - beta-adrenergic receptor pharmacogenetics. PMID- 19804214 TI - Study fails to validate genetic risk factors for ACS. PMID- 19804215 TI - Double jeopardy for the mentally ill: higher cardiovascular risk and reduced frequency of certain interventional procedures. AB - Evaluation of: Kisely S, Smith M, Lawrence D et al.: Inequitable access for mentally ill patients to some medically necessary procedures. CMAJ 176[6], 779 784 [2007]. This observational database study utilized data from three administrative databases to determine the relative effect of psychiatric disease status on mortality and utilization of in-patient procedures for cardiovascular conditions. The study results indicated that psychiatric patients exhibit significantly greater mortality compared with other groups of patients, despite adjustments for relevant socioeconomic and clinical factors. Compounding the problem, the authors further determined that cardiovascular medical procedures were underutilized in this population. This important study reveals significant public health implications for a patient population already at elevated risk for cardiovascular morbidity and mortality due to lifestyle, medical comorbidities and certain antipsychotic therapies. This study now offers supportive evidence that inequities in primary care health delivery systems provide a partial explanation for the excess cardiovascular risk observed in mentally ill patients. The widespread recognition of this issue should prompt additional research and consequent remedial action by medical providers and those institutions that support them. PMID- 19804216 TI - Can intense exercise contribute to cardiovascular disease? AB - Aerobic exercise is associated with a number of beneficial effects, including reduction in cardiovascular morbidity and mortality, decreased blood pressure, improved insulin sensitivity, weight loss and favorable alterations in lipids. However, intense exercise may have significant deleterious cardiovascular effects. Excessive physical activity may cause increased oxidative stress with resultant endothelial dysfunction and progressive atherosclerosis. Furthermore, intense exercise may promote exercise-induced hypertension that is also associated with endothelial dysfunction and increased cardiovascular mortality. The purpose of this article is to summarize our current understanding of whether intense exercise can contribute to the development of cardiovascular disease. PMID- 19804217 TI - Validation of the human tissue factor/FVIIa complex as an antithrombotic target and the discovery of a synthetic peptide. AB - This review focuses on the validation of the principal initiator of human coagulation, the tissue factor (TF)/coagulation factor (F)VIIa complex, as an antithrombotic target, as well as on the discovery of a cyclic pentapeptide (PN7051), which dose-dependently inhibits TF/FVIIa-induced coagulation and thrombus formation. Target validation and studies of antithrombotic efficacy were performed with a human thrombosis model employing non-anticoagulated blood from severe homozygous FVII-deficient patients and healthy individuals at blood-flow conditions mimicking those in healthy and diseased vessels. Additional validation included an anti-TF monoclonal antibody, recombinant TF pathway inhibitor, recombinant inactivated-active site FVIIa and all-trans retinoic acid. Structural and biological characterization of PN7051 and other peptides from the same FVII domain indicate that PN7051 interferes with an essential interaction between the epidermal growth factor domain-2-like and the catalytic domains of FVIIa. A peptidomimetics approach is suggested to further improve the antithrombotic potency of PN7051. PMID- 19804218 TI - gamma-glutamyltransferase and pathogenesis of cardiovascular diseases. AB - Cardiovascular epidemiology has recently highlighted a clear link between serum gamma-glutamyltransferase (GGT) and risk for stroke, infarction and cardiovascular death, associated with the evolution of atherosclerosis-related conditions, such as coronary artery and cerebrovascular disease. Thus, serum GGT is now recognized as a cardiovascular prognostic marker. However, the reasons for the association between serum GGT elevations and unfavorable prognosis remain to be defined. Histochemistry and biochemistry have provided intriguing clues, focusing new research approaches on the complex issue of monitoring and treatment of chronic cardiovascular conditions. PMID- 19804219 TI - Visceral debranching to facilitate repair of complex thoracic and thoracoabdominal aneurysms. AB - The availability of thoracic and abdominal endografts has decreased the morbidity and mortality associated with the repair of aneurysms of the aorta. The current limitations of endografts for aneurysms involve adequate proximal and distal landing zones. Branches that are not critical, such as the internal iliac or left subclavian artery, may often be covered and included in the landing zone for most patients, with a few exceptions. However, aneurysms involving critical branch vessels to the brain and intra-abdominal viscera cannot be covered and open repair of aneurysms in these locations carry the highest morbidity and mortality. This review describes the various techniques to relocate these critical branch vessels to expand the use of stent grafts for the treatment of these complex aneurysms. PMID- 19804220 TI - Sex hormones, vascular function and the outcome of hormone replacement therapy in cardiovascular disease. AB - Cardiovascular disease is more common in men and post-menopausal women than premenopausal women, suggesting that female sex hormones have vascular benefits. Cytosolic/nuclear estrogen and progesterone receptors mediate genomic transcriptional effects that stimulate endothelial cell growth and inhibit smooth muscle proliferation. Sex hormone receptors on the plasma membrane trigger nongenomic stimulation of endothelium-dependent nitric oxide-cyclic (c)GMP, prostacyclin-cAMP and hyperpolarizing vascular relaxation pathways, as well as inhibition of [Ca2+](i), protein kinase C and Rho-kinase-dependent mechanisms of smooth muscle contraction. Despite the vasodilator effects of sex hormones, the Heart and Estrogen/progestin Replacement Study (HERS), HERS-II and Women's Health Initiative clinical trials have shown minimal benefits of hormone replacement therapy (HRT) in post-menopausal cardiovascular disease. The prospect of HRT relies on further mechanistic analysis of the vascular effects of natural sex hormones and phytoestrogens, and the identification of specific estrogen receptor modulators. Androgens have vascular effects, and modulators of the estrogen/testosterone ratio could provide better HRT combinations. The timing/duration and the type, dose and route of administration of HRT should be customized according to the subject's age and pre-existing cardiovascular condition, thereby enhancing the outcome of HRT in cardiovascular disease. PMID- 19804221 TI - Anticoagulant therapy in acute coronary syndromes. AB - Thrombus formation on a disrupted atherosclerotic coronary artery plaque is the usual event that precipitates acute coronary syndromes (ACS). Anticoagulation therapy with unfractionated heparin, in addition to antiplatelet therapy with aspirin, decreases the risk of myocardial infarction and death with ACS. However, unfractionated heparin has pharmacological limitations that limit efficacy and safety. Low-molecular-weight heparins (enoxaparin), direct thrombin inhibitors (bivalirudin) and Factor Xa inhibitors (fondaparinux) are new anticoagulant therapy options with either superior efficacy or improved safety over unfractionated heparin. Compared with unfractionated heparin, enoxaparin and fondaparinux are easier to administer, do not require monitoring and facilitate longer treatment duration. Bivalirudin offers advantages for patients undergoing percutaneous revascularization. Careful attention to dosing and excellent vascular access site management are required to decrease the risk of bleeding and blood transfusion, which have been associated with increased mortality risk. PMID- 19804222 TI - Oxidation hypothesis of atherogenesis: HDL inflammatory index and apolipoprotein A-I mimetic peptides. AB - Coronary heart disease is the leading cause of death in the USA and worldwide. Optimizing the ratio and levels of low- and high-density lipoproteins (HDLs) has been a major focus of treatments in preventing atherosclerosis. While these therapies have made significant contributions in reducing heart disease, many patients with normal lipid levels still continue to have significant coronary heart disease and crippling cardiac events. Recent research has brought to light the fact that HDL, widely touted to be protective against atherosclerosis, can actually promote atherogenesis in certain conditions. Disruption of anti inflammatory properties of HDL may result in atherosclerosis in the presence of decreased, increased or unchanged serum HDL-cholesterol levels. The ability of HDL to prevent or promote atherogenesis can be assessed using a parameter termed the HDL inflammatory index. The next generation of emerging therapeutics, such as apolipoprotein A-I mimetic peptides, will be aimed at improving the anti inflammatory property of HDL and thus further reducing the incidence of coronary heart disease. PMID- 19804224 TI - Bringing cardiac stem cell therapy into the clinic. PMID- 19804223 TI - Vitamin K antagonist use in the elderly: special considerations. AB - As the population ages, the number of patients aged 75 years and over treated with vitamin K antagonists (VKAs) is steadily increasing. In this age group, the two main indications for oral anticoagulant therapy are the treatment of venous thromboembolic disease and the prevention of systemic embolism in patients with nonvalvular atrial fibrillation. In both indications, a target international normalized ratio of 2.5 (range: 2.0-3.0) is recommended. Although VKAs are beneficial in thromboembolic disorders, they are still underused. In this review, we will focus on two crucial topics in elderly patients, the specific management of VKAs in these patients and the hemorrhagic risk. Current recommendations concerning the management at the start of treatment, education and adequate monitoring may help to minimize the hemorrhagic risk in these frail patients. PMID- 19804225 TI - Space technology being used to detect and monitor heart conditions. PMID- 19804226 TI - Antithrombotic strategies in non-ST elevation acute coronary syndromes: focus on bivalirudin. AB - Acute coronary syndromes (ACS) are a common presentation of coronary artery disease, accounting for more than one million hospital admissions in the US annually. Owing to high rates of mortality and reinfarction, ACS represent a major public health concern. The following review discusses the pathogenesis of ACS and optimal approaches for the management of patients with ACS, with special focus on new antithrombotic strategies, including the direct thrombin inhibitor bivalirudin. Bivalirudin has several notable mechanistic advantages compared with unfractionated heparin, including activity against clot-bound thrombin, inhibition of thrombin-induced platelet activation, short plasma half-life in patients with normal or mildly impaired renal function (25 minutes), and linear pharmacokinetics less affected by plasma proteins and renal insufficiency. These properties provide a more predictable inhibition of coagulant activity than unfractionated heparin, with less degree of inter-patient variability in anticoagulation response. The findings from the several clinical trials assessing safety and efficacy of bivalirudin are analyzed in detail, including the recent randomized controlled Acute Catheterization and Urgent Intervention Triage Strategy (ACUITY) trial. Based on the results of the ACUITY trial, a newer streamlined strategy for the invasive treatment of moderate- and high-risk patients with ACS is discussed. PMID- 19804227 TI - Recombinant nematode anticoagulant protein c2 in non-ST segment elevation acute coronary syndrome and beyond. AB - Patients with acute coronary syndromes (ACS) have high recurrent ischemic event rates despite management with current guideline-based therapies. Recombinant nematode anticoagulant protein (rNAP)c2 provides factor Xa-dependent inhibition of the tissue factor/factor VIIa complex acting proximally on the clotting cascade. It may be administered either intravenously or subcutaneously and has an elimination half-life of approximately 50-60 h. rNAPc2 reduces thrombin formation in patients undergoing elective percutaneous coronary interventions (PCI) and in patients with non-ST segment elevation ACS managed with an early invasive strategy, while bleeding rates are comparable with currently used anticoagulants. Patients receiving rNAPc2 undergoing emergent coronary artery bypass surgery within 96 h of dosing have increased rates of major bleeding. Some heparin coadministration may be necessary to avoid PCI-related thrombotic complications. Large-scale trials are needed to confirm these findings and to evaluate the impact of rNAPc2 on clinical events. PMID- 19804228 TI - Sitaxsentan for the treatment of pulmonary arterial hypertension. AB - Pulmonary arterial hypertension (PAH) is a life-threatening, progressive condition that eventually leads to right heart failure. Endothelin receptor antagonists (ETRAs) have been shown to be a safe and efficacious treatment for PAH. Sitaxsentan is a highly selective oral ETRA that, in clinical trials, has demonstrated significant improvements in exercise capacity, functional class and pulmonary hemodynamics in PAH. Sitaxsentan has been shown to be a well tolerated and effective agent with a durable response for the treatment of PAH, and is associated with a low incidence of liver toxicity. PMID- 19804229 TI - Role of p38MAPK in beta(2)AR-induced cardiomyopathy: at the heart of the matter? AB - Peter PS, Brady JE, Yan L et al.: Inhibition of p38alpha MAPK rescues cardiomyopathy induced by overexpressed beta(2)-adrenergic receptor, but not beta(1)-adrenergic receptor. J. Clin. Invest. 117(5), 1335-1343 (2007). The role of p38MAPK in the progression of heart failure is controversial. In the reference paper, Peter and colleagues demonstrated that p38MAPK plays an important role in the progression of heart failure in transgenic mice over-expressing the beta(2) adrenergic receptor. The authors show that p38MAPK is involved in apoptosis, fibrosis and decreased left ventricle fractional shortening in mice overexpressing beta(2) adrenergic receptor but not beta(1) adrenergic receptor. These results showed that beta(1) and beta(2) adrenergic receptors induce cardiomyopathies through different signaling pathways, and suggest an important role for p38MAPK in the development of cardiac disease. PMID- 19804230 TI - Platelet transcriptome and cardiovascular disease. AB - Platelet hyper-reactivity is likely to play a role in cardiovascular disease, but there are no standardized tests to evaluate platelet responsiveness. A 'platelet chip' (a synthetic oligonucleotide microarray representing all platelet restricted genes) is under development as a tool for high-throughput characterization of platelet-based bleeding and clotting disorders. In future, platelet gene profiling may be used to improve thrombohemorrhagic risk assessment and to guide antiplatelet therapy for patients at risk of cardiovascular disease. PMID- 19804231 TI - Role of interventional repair in the thoracic aorta. AB - Endovascular treatment of chronic aneurysmatic diseases of the descending thoracic aorta has demonstrated encouraging peri-interventional mortality and morbidity and is accepted as a preferred strategy in experienced centers. The emergence of endovascular strategies for acute thoracic aortic pathologies is an even more exciting new territory for nonsurgical interventions considering the sobering results of open surgery. Although it is apparent that patients at high risk for open surgery will benefit from endovascular strategies, the exact role of stent-graft placement remains to be defined, as the community awaits solid long-term data and as devices and techniques continue to improve. While some indications and scenarios, such as acute type B dissection with associated malperfusion syndrome or imminent aortic rupture, have been shown to benefit from stent-graft treatment, others are less settled. The current paper discusses both the established and emerging indications, as well as technical and anatomical aspects of this fascinating therapeutic option. PMID- 19804232 TI - Treatment of heart failure by calcium cycling gene therapy. AB - Heart failure is a major cause of morbidity and mortality in Western countries. While progress in conventional treatment modalities is making steady and incremental gains to reduce this disease burden, there remains a need to explore new and potentially therapeutic approaches. Gene therapy, for example, was initially envisioned as a treatment strategy for inherited monogenic disorders. It is now apparent that gene therapy has broader potential, which also includes acquired polygenic diseases such as heart failure. Advances in the understanding of the molecular basis of conditions such as these, together with the evolution of increasingly efficient gene transfer technology, has placed congestive heart failure within the reach of gene-based therapy. PMID- 19804233 TI - Tissue engineering approaches for the development of a contractile cardiac patch. AB - Cardiac tissue engineering is an emerging field that aims to enhance cardiac function in patients with heart disease or congenital malformations, by providing suitable implantable replacement tissues. Various groups have developed methods of cultivating contractile 3D cardiac grafts that display functional and morphological properties of native myocardium, integrate into native tissue and provide improvement in cardiac function upon grafting onto infarcted hearts. However, several critical goals must be met before this therapy can be implemented clinically: the identification and harvesting of an appropriate human cell source, the cultivation of relatively thick ( approximately 1 cm) constructs, and the in vitro and/or in vivo vascularization of these constructs. Advances in enabling bioreactor technologies and the identification of key parameters governing the differentiation of cardiac cells will be crucial to advancement in this field. PMID- 19804234 TI - Use of predictive markers to improve cardiovascular protection. AB - Together with other modifiable cardiovascular risk factors, hypertension heavily contributes to the global burden of cardiovascular morbidity and mortality, as well as to the increase in individual absolute cardiovascular risk. Comparison of the effectiveness of different therapies in reducing the incidence of major cardiovascular events has classically required the evaluation of major 'hard' end points. In view of the long natural history of hypertension, however, it appears very useful to monitor modifications in measurable 'intermediate' end points or 'disease markers'. This approach may provide more accurate individual risk stratification and a better evaluation of the efficacy of a given treatment in preventing or modifying the course of target organ damage. This may represent a valuable and affordable strategy in clinical practice allowing the evaluation of both patient prognosis and the effectiveness of antihypertensive treatment over time. PMID- 19804236 TI - Brain natriuretic peptide: does it play a role in post-cardiac surgery patients? PMID- 19804235 TI - Effect of smoking on abdominal aortic aneurysms: novel insights through murine models. AB - Abdominal aortic aneurysm is a poorly understood and fatal disease. The etiology of the disease is believed to be multifactorial. Of all the recognized clinical associations, none has a greater impact on the incidence and progression of the disease than exposure to tobacco smoke. Novel murine models developed over the past several years present the opportunity to investigate the mechanism of this critical clinical relationship. PMID- 19804237 TI - Future strategies for improving outcomes in patients with diabetes and acute coronary syndromes. PMID- 19804238 TI - Adipose stem cells used in heart failure treatment. PMID- 19804239 TI - Atorvastatin for reduction of myocardial dysrhythmia after cardiac surgery study. AB - Evaluation of: Patti G, Chello M, Candura D et al.: Atorvastatin for Reduction of MYocardial Dysrhythmia After cardiac surgery (ARMYDA-3) study. Circulation 114, 1455-1461 (2006). The Atorvastatin for Reduction of MYocardial Dysrhythmia After cardiac surgery study evaluated the efficacy of atorvastatin 40 mg/day compared with placebo, adminstered 1 week prior to surgery in preventing post-operative atrial fibrillation (AF). The rate of post-operative AF and mean post-operative hospital stay was significantly lower in the atorvastatin group compared with the placebo group. The incidence of the composite end point at 1 month was similar in both groups. This study supports the initiation of atorvastatin in patients undergoing cardiac surgery to reduce the rate of post-operative AF and the duration of hospital stay. Future studies will be required to assess whether this beneficial effect of atorvastatin is demonstrated by all statins and whether it can be extended to the wider population of patients with AF. PMID- 19804240 TI - Metabolic perturbation in chronic heart failure: time to redress the balance? AB - Evaluation of: Jankowska EA, Biel B, Majda J et al. : Anabolic deficiency in men with chronic heart failure; prevalence and detrimental impact on survival. Circulation 114, 1829-1837 (2006). The study examined the circulating levels of three anabolic hormones (testosterone, insulin-like growth factor-1 and dehydroepiandrosterone sulfate) in 208 men with stable chronic heart failure and 366 healthy controls. A high prevalence of anabolic hormone deficiency (89% deficient in one or more hormone) was found in heart failure patients. Hormone levels were weakly related to disease severity and hormone deficiency independently predicted prognosis (a 3-year survival rate of 27% was found in subjects deficient in all three hormones). Although it cannot be concluded that anabolic hormone deficiency contributes to the disease process - it may simply reflect chronic disease with no physiological consequence - it is highly plausible that deficiency may at least contribute to symptomatology. Further studies evaluating anabolic hormone replacement for symptomatic improvement and possible prognostic benefit are warranted. PMID- 19804241 TI - REPAIR-AMI: stem cells for acute myocardial infarction. AB - Evaluation of: Schachinger V, Erbs S, Elsasser A et al.: Intracoronary bone marrow-derived progenitor cells in acute myocardial infarction. N. Engl. J. Med. 355, 1210-1221 (2006). The Reinfusion of Enriched Progenitor cells And Infarct Remodeling in Acute Myocardial Infarction (REPAIR-AMI) trial, the largest randomized, placebo-controlled trial of stem cell therapy in acute myocardial infarction, studied the efficacy of the intracoronary delivery of bone marrow mononuclear cells (BMCs) versus placebo in patients with acute ST-segment elevation myocardial infarction following successful percutaneous coronary intervention. At 4 month follow-up, patients treated with BMCs had a significant improvement in left ventricular ejection fraction compared with placebo (+5.5 vs +3.0%, absolute difference +2.5%). In addition, treatment with BMCs was associated with a statistically significant reduction in adverse clinical events at 1 year follow-up. Despite these promising findings, other studies have shown mixed results and several unresolved clinical and physiological issues remain. Key findings from ongoing basic and clinical research will define the future role of stem cell therapy for acute myocardial infarction. PMID- 19804242 TI - Strategy of transfusion in cardiac surgery: limits of hematocrit and how much is too low? AB - The use of blood and blood products in cardiac surgery is higher than necessary and a reduction is imperative due to complications and costs. Hemodilution is unavoidable in cardiopulmonary bypass and is the most likely pitfall when evaluating transfusion needs. Even patients with coexisting cardiovascular diseases tolerate perioperative hemodilution better than most anticipate. Hemodynamic monitoring is important to evaluate the association between hemoglobin level and organ function. Use of both mechanical and medical blood conservation strategies is required to reduce blood transfusion, and most of the methods have a positive cost-effectiveness and cost-benefit. By using the right strategy and policy, transfusion of blood and blood products can be reduced to less than 5% of cardiac patients. PMID- 19804243 TI - Is post-traumatic stress disorder related to development of heart disease? AB - It has long been hypothesized that post-traumatic stress disorder (PTSD) increases coronary heart disease (CHD) risk, however, empirical evidence is limited. In the first prospective study to date, individuals with higher PTSD symptom levels had a significantly increased risk for CHD, after controlling for known coronary risk factors. PTSD indicates a chronic stress reaction and is hypothesized to influence CHD either by causing biological alterations that lead to cardiovascular damage, or by leading to adverse health behaviors that increase CHD risk. A key issue is whether PTSD contributes to the development of CHD, if PTSD and CHD share common pathways or if CHD causes PTSD. Research combined across different disciplines suggests that prolonged or chronic stress does influence the development of CHD. A better understanding of the relationship will increase prevention and intervention efforts. Cardiologists may be most effective when they can recognize and manage emotional distress in practice. PMID- 19804244 TI - Impact of statins on atherosclerotic plaque. AB - Lowering levels of low-density lipoprotein cholesterol with inhibitors of 3 hydroxy-methylglutaryl coenzyme A reductase (statins) has a profound impact on cardiovascular event rates. The degree of benefit is proportional to the extent of low-density lipoprotein cholesterol lowering achieved. In addition to lowering levels of atherogenic lipids, statins have been reported to possess nonlipid modifying properties that may contribute to their clinical benefit. It remains to be determined to what extent these properties modify atherosclerotic plaque in humans. A number of imaging modalities provide a unique opportunity to monitor the effect of medical therapies on the extent and composition of atherosclerotic plaque. The use of these imaging modalities highlights the impact that statins have on atherosclerosis as a possible rationale for their clinical benefit. PMID- 19804245 TI - Advances in the treatment of acute decompensated heart failure in the elderly. AB - Acute decompensated heart failure is the leading cause of hospitalization in older adults, and more than half of all patients admitted with this condition are over 75 years of age. In addition, hospital mortality is threefold higher in patients over 75 years of age compared with younger patients. This article reviews the pathophysiology, clinical features and management of acute heart failure in older adults, highlighting recent advances in the field. It is anticipated that over the next 5-10 years, new approaches to the treatment of acute decompensated heart failure will become available. Nonetheless, additional research is required to develop more effective strategies for the prevention and management of both acute and chronic heart failure in our rapidly growing elderly population. PMID- 19804246 TI - Left atrial stiffness and its implications for cardiac function. AB - Proper atrial function is essential for overall cardiovascular performance, mainly by its four major functions, namely, bioelectrical, hormonal, metabolic and hemodynamic. With regard to the latter, atria modulate ventricular filling by smoothing the transformation of the continuous venous return to the intermittent filling pattern of the ventricles during diastole through three main components: a phase of reservoir mainly during ventricular systole, a conduit phase during ventricular diastole and an active phase in late ventricular diastole. Although the atria assume a dynamic role in ventricular filling progression, atrial function and emptying pattern is, conversely, highly influenced by the ventricular diastolic wall stress, underlying the close connection observed between these chambers. This review focuses essentially on left atrial mechanical role, particularly on the physiological and clinical consequences of disturbed atrial compliance. PMID- 19804247 TI - Use of intermittent pneumatic compression in the treatment of venous ulcers. AB - Venous ulcers affect up to 1% of the population. This review provides the hemodynamic, hematologic and clinical effects of intermittent pneumatic compression (IPC) in the treatment of venous ulcers. It presents the different IPCs and their application on venous disease. It points out that a large randomized study on IPC versus standard four-layer compression in the treatment of venous ulcers is needed. The cost-effectiveness of IPC in the treatment of venous ulcers should also be assessed. PMID- 19804248 TI - Thrombus formation on apex of arterial stenoses: the need for a fluid high shear stenosis diagnostic device. AB - This review is focused upon the studies of thrombus formation in human non anticoagulated blood on an apex of an eccentric stenosis positioned in the blood flow channel of a parallel-plate perfusion chamber. Thrombus formation in blood from healthy individuals and patients with various bleeding disorders, as well as the effects of a diet supplement and pharmacological interventions, are discussed in view of thrombus-forming mechanisms under these complex blood-flow conditions. Hallmarks of this significantly enhanced thrombus formation are the apparent dependence on thrombin generation, shear-induced platelet activation, induction of platelet procoagulant activity and pronounced platelet microparticle formation that parallel the growth of these fibrin-rich thrombi. The development of miniature models of these blood-flow devices for diagnostic purposes is suggested for the assessment and monitoring of the efficacy of antithrombotic regimens in blood from patients with atherosclerotic disease in parallel with assessments of platelet microparticle formation, shear-induced platelet activation and platelet procoagulant activity. PMID- 19804249 TI - Effects of caffeine and coffee consumption on cardiovascular disease and risk factors. AB - Clinical and observational studies have recently reported that coffee consumption is associated with cardiac arrhythmia, with increased serum cholesterol, blood pressure and affected heart rate variability, leading to an increased cardiovascular risk. Analysis of these papers shows that these data are controversial and strongly depend on methodology. Recent reports demonstrate the beneficial effects of coffee consumption due to anti-inflammatory actions mediated by antioxidant compound of the beverages. PMID- 19804250 TI - Mechanical and pharmacological methods to prevent venous thromboembolism. AB - Today, it is possible to define certain risk groups for the development of venous thromboembolism, most studies having been conducted on post-operative venous thromboembolism. Nonetheless, the risk classification is rather inexact, resulting in many patients receiving prophylaxis who would not have gone on to develop the complication. However, the current prophylactic methods are safe and can therefore also be used in the post-operative situation. The dominating pharmacological substances are unfractionated heparin, low-molecular-weight heparins and the pentasaccharide fondaparinux. In some situations (such as elective hip surgery and likely hip fracture surgery, and in many cases where malignant diseases in the abdomen/pelvis have been operated upon), extended prophylaxis for 1 month should be considered. PMID- 19804251 TI - Cardiac natriuretic hormones as clinical markers: instructions for use. PMID- 19804253 TI - Tolvaptan for the treatment of hyponatremia and congestive heart failure. AB - Tolvaptan is an oral, once-daily nonpeptide arginine vasopressin V(2)-receptor antagonist under development for the treatment of hyponatremia and congestive heart failure. In Phase II clinical trials, tolvaptan, in addition to standard therapy, increased fluid loss, resulting in decreased body weight and improved edema and serum sodium without affecting blood pressure, heart rate or renal function in patients with heart failure. The compound appeared to be well tolerated and dose-dependent adverse events were generally realated to its pharmacological activity, such as thirst and dry mouth. In patients with hyponatremia, tolvaptan appears to be more effective than fluid restriction at improving sodium levels without an increase in adverse events. An international Phase III outcome study; Efficacy of Vasopressin antagonism in hEaRt failurE outcome Study with Tolvaptan (EVEREST), evaluating the long-term efficacy and safety of tolvaptan in patients hospitalized with worsening heart failure, is currently ongoing. PMID- 19804254 TI - Micronized fenofibrate: a useful choice for the correction of dyslipidemia in metabolic syndrome and Type 2 diabetes. AB - Cardiovascular disease is the principal cause of illness and disability in patients with diabetes, and is also the most common cause of death worldwide in adults. Fenofibrate, a member of the fibrate class of lipid-modifying drugs, is a potent triglyceride-lowering and high-density lipoprotein cholesterol-raising agent and has a variable effect on low-density lipoprotein cholesterol. Fenofibrate administration also leads to a modified, less atherogenic low-density lipoprotein profile, with a consistent effect toward increased low-density lipoprotein particle size and a reduction in the low-density lipoprotein particle density. Maximal clinical efficacy in fibrates has been demonstrated in subjects with dyslipidemia, particularly in populations with features of the metabolic syndrome and in patients with Type 2 diabetes. Angiographic data from the Diabetes Atherosclerosis Intervention Study (DAIS) support a similar effect of fenofibrate. However, in the recent Fenofibrate Intervention and Event Lowering in Diabetes trial (FIELD; 9795 patients with Type 2 diabetes), the rate of nonfatal macrovascular events, after adjustment for the use of other lipid lowering agents and significant reductions in microvascular complications, was lower for the fenofibrate treatment group. These results and those from a current large trial, ACtion to COntrol cardiovascular Risk in Diabetes (ACCORD), will provide valuable evidence for the likely future use of this drug in combination with statins for reducing cardiovascular disease risk in the metabolic syndrome and in Type 2 diabetes. PMID- 19804255 TI - Multidisciplinary congress in cardio-thoracic healthcare. PMID- 19804256 TI - The ASTEROID trial: coronary plaque regression with high-dose statin therapy. AB - A Study To Evaluate the effect of Rosuvastatin On Intravascular ultrasound Derived coronary atheroma burden (ASTEROID) investigated the impact of high-dose rosuvastatin therapy on the rate of atheroma progression in patients with coronary artery disease. Serial intravascular ultrasound (IVUS) was performed in 349 patients at baseline and following 24 months of therapy with rosuvastatin 40 mg/day. Rosuvastatin therapy lowered low-density lipoprotein cholesterol to 60.8 mg/dl and raised high-density lipoprotein cholesterol by 14.7%. This was associated with a significant reduction in all IVUS measures of atheroma burden. These results suggest that intensive modification of lipid levels with high-dose statin therapy can promote atheroma regression. Further studies will be required to determine whether this benefit is associated with a reduction in clinical events. PMID- 19804257 TI - Can vascular closure devices reduce complications? AB - Vascular closure devices have demonstrated their ability to reduce hemostasis and ambulation times, as well as improve patient comfort, without increasing vascular complications in their initial industry-sponsored, randomized clinical trials. However, in the early years of first-generation closure devices, there was an initial concern about the risk of higher complication rates. As devices have improved, it appears that there may now be a trend towards reduced vascular complications compared with manual compression, especially in patients undergoing percutaneous coronary interventions. PMID- 19804258 TI - Is there potential for antioxidants to enhance thrombolysis therapy in patients with ischemic stroke? AB - The medical and socio-economic burden of ischemic stroke is vast. Current thrombolytic therapies have a time-limited therapeutic window and do not provide significant benefits beyond tissue reperfusion. The detrimental effect of oxidative stress caused by excessive oxidant production due to cerebral reperfusion injury is a neglected consequence of ischemic stroke and warrants special consideration. Strategies directed at preventing or reducing oxidative damage in the brain post-ischemic stroke have the potential to improve neurological outcome and reduce morbidity and mortality from this common disease. Significantly, the prospect of increasing the size of the treatment window for thrombolytic therapies, perhaps by synergistic effects with other medications given in parallel, is also an avenue worthy of further investigation. This perspective outlines the current status of thrombolytic therapy for the treatment of ischemic stroke and explores the possibility of improving and expanding this potential therapy. Furthermore, the implications of directly treating damage caused by oxidative stress with novel antioxidant therapy are discussed. PMID- 19804259 TI - Natriuretic peptides: diagnostic tools and predictors of heart failure outcome. AB - Congestive heart failure (CHF) is a progressive disease whose outcome largely depends on early, accurate and prompt diagnosis, accompanied by evidence-based treatment. The explosion of uptake of natriuretic peptides (NPs) in clinical practice belies an understanding of how peptides are used. The signal for NP release is the same signal that causes symptoms of CHF, such as increased wall stress. Thus, NPs can reliably add to the information a physician brings to the table as they attempt to diagnose the acutely dyspneic patient with CHF. Additionally, NPs have strong prognostic utility in the emergency room and the hospital. Monitoring of NPs during treatment for acute CHF may help manage the patient. In the future, it is possible that NPs will play a more prominent role in early detection of left ventricular dysfunction as well as guiding chronic CHF treatment. PMID- 19804260 TI - Hormone therapy and cardiovascular risk markers and disease: focus on progestagens. AB - Biological studies have demonstrated estrogen's beneficial effect on cardiovascular risk factors, including plasma lipoproteins, atherogenesis, vascular reactivity, inflammation and antioxidative activity. Additionally, observational studies have supported a cardioprotective effect of hormone therapy (HT), although an underlying healthy-user effect may account for these observations. Progestagens are added to protect against an increased risk of endometrial cancer observed with unopposed estrogen treatment. The inclusion of progestagen in HT has been associated with possible adverse cardiovascular outcomes. Recent, large-scale, randomized clinical studies did not confirm a beneficial cardiovascular effect of HT. On the contrary, an increased risk was found with continuous combined estrogen-progestagen regimens. The progestagen used in these trials was medroxyprogesterone acetate and other progestagen components have only been sparsely elucidated. The purpose of the present review is to outline some of the modifying effects of different progestagens on the actions of estrogen on cardiovascular risk markers and clinical end points observed in biological, observational and clinical studies. PMID- 19804261 TI - Potential role of statin therapy in heart failure, atrial fibrillation and aortic stenosis. AB - 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors, or statins, are widely prescribed throughout the world, and considerable evidence has indicated their powerful effects in ischemic forms of cardiovascular disease. Recently, several trials have demonstrated that statins have pleiotropic effects beyond their lipid-lowering capacities. These findings may play a role in the use of statins to manage forms of cardiovascular disease that may or may not have an ischemic etiology: congestive heart failure, atrial fibrillation and aortic stenosis. PMID- 19804262 TI - Vena cava filters: an update. AB - Venous thromboembolism is a major health problem that results in significant long term complications and mortality. The management of venous thromboembolism is complex and can be particularly challenging when pharmacological therapy alone cannot be effectively utilized. Vena cava filters provide protection from pulmonary embolism for patients in whom therapeutic anticoagulation is contraindicated or inadequate. Recent innovations in caval interruption have included the use of alternative imaging modalities for filter insertion and the emergence of devices designed to allow temporary caval filtration. These developments have been accompanied by a controversial increase in the use of vena cava filters for prophylactic indications in the absence of venous thromboembolism. In addition to a brief historical perspective on caval filtration, this update reviews the indications for vena cava filter insertion, associated complications, methods of caval imaging and filter insertion and current FDA-approved devices. PMID- 19804263 TI - Advanced wiring technique for chronic coronary total occlusions. AB - Percutaneous coronary intervention (PCI) of chronic total occlusion (CTO) is one of the greatest challenges in interventional cardiology. The procedural success rate is relatively low, mainly due to guidewire failure to cross the occlusion. An effective wiring technique is the key to the success of CTO-PCIs. However, success or failure of wire crossing is mostly dependent on an operator's experience and skill. To improve the success rate of CTO-PCI in current interventional cardiology, several techniques were introduced in the Japanese CTO club. In this article, some advanced wiring techniques developed by the Japanese CTO club will be explored. PMID- 19804264 TI - Clinical implications of vulnerable plaque. AB - In many individuals, the first indicator of atherosclerosis is an acute heart attack, which is often fatal. Despite innovations in medical therapy and interventional cardiology techniques, coronary artery disease continues to be the leading cause of death in the USA. There is great interest in identifying vulnerable plaques and vulnerable patients as a possible means to stem the tide against coronary artery disease. Improvements in diagnostic studies and development of novel imaging tools have opened the possibilities for significant advances in the management of vulnerable plaque. The result of improved risk stratification, by both noninvasive and invasive means, will be a better assessment of the risk/benefit relationships for the novel therapies that are needed to further reduce the morbidity and mortality of the disease. Correct identification of vulnerable plaque would permit the use of more effective systemic treatment and enable clinical trials to study the supplemental benefit from local treatments. PMID- 19804266 TI - Understanding how statins work: the path to better treatments for heart disease and more. PMID- 19804265 TI - Integrating principles of developmental biology in tissue engineering of heart valves. PMID- 19804267 TI - Have the risks of rosiglitazone been exaggerated? PMID- 19804268 TI - MCP-1 found to predict prognosis in acute and chronic phases after acute coronary syndrome. PMID- 19804269 TI - Post-menopausal hormone therapy and cardiovascular events: a new paradigm based on pharmacogenetics? AB - The risk of coronary heart disease (CHD) events increases in women after menopause and it was thought that hormone therapy (HT) would decrease this risk. However, numerous large randomized clinical trials have not demonstrated decreased risk in secondary prevention and some have actually shown increased risk of cardiovascular events with HT in primary prevention. Platelets are involved in CHD events and platelet glycoprotein single-nucleotide polymorphisms may help to identify risk and treatment efficacy. It may become a plausible strategy in the future for clinicians to identify, by genotyping, high-risk patients who would benefit from HT to reduce CHD risk as well as to identify patients with neutral risk for whom HT could be harmful. PMID- 19804270 TI - Takotsubo cardiomyopathy. AB - Takotsubo cardiomyopathy is a disease in which patients present with signs and symptoms mimicking an acute coronary syndrome and, thus, undergo cardiac catheterization. At the time of catheterization, however, no critical coronary lesions are found. Most of these patients have a characteristic left ventriculogram and recent history of a preceding stressor. While the acute phase of the illness can lead to ventricular tachyarrhythmias and possibly even death, a key feature of this syndrome in the majority of patients is its generally benign course and reversibility in myocardial dysfunction over a period of days to weeks. The pathophysiology of Takotsubo cardiomyopathy remains unclear. Optimal long-term medical therapy has not been well established at this time, since the etiology of the disease process remains a source of debate. PMID- 19804271 TI - Monitoring antiplatelet therapy with point-of-care platelet function assays: a review of the evidence. AB - Multiple studies have demonstrated that subgroups of patients receiving combination therapy with aspirin and clopidogrel fail to produce the anticipated antiplatelet effect, and various terms such as 'aspirin resistance', 'clopidogrel resistance', 'heightened post-treatment platelet reactivity' and 'residual platelet reactivity' have been introduced in the medical literature. Light transmittance aggregometry is generally considered to be the gold standard for determining platelet function, but its relevance to in vivo platelet function is questionable and the logistical demands of the method make it impossible to use in daily practice. The introduction of several point-of-care platelet function assays may be the key to the widespread clinical use of platelet function testing and may identify patients who are at risk for the occurrence of adverse cardiac events. In the present paper, we discuss the current commercially available methods of assaying platelet function, including their advantages and limitations and whether they have been shown to correlate with clinical outcomes. PMID- 19804272 TI - Heart disease and erythropoietin. AB - An additional target for reducing infarct size, namely, attenuation of apoptosis, has recently emerged. Erythropoietin (Epo) exhibits properties that may attenuate this process and enhance neovascularization, thereby preserving jeopardized myocardium. Potentially adverse effects of Epo, including hypertension, thrombosis and possible exacerbation of occult neoplasms can likely be averted with analogues such as carbamylated and asialo Epo, which are devoid of erythropoietic effects, yet retain tissue-protective characteristics. With a single, but adequate dose of Epo administered early after the onset of acute myocardial infarction, coupled with therapy to induce reperfusion, tissue protection conferred by Epo and its analogues may facilitate the preservation of myocardium subjected to ischemic insults, thereby improving prognosis. PMID- 19804273 TI - Future strategies to prevent renal microvascular disease complications in diabetes. AB - Nephropathy is one of the major chronic microvascular complications of diabetes. The epidemic of Type 2 diabetes and related incidence of end-stage renal disease is progressively increasing worldwide and represents a major public health concern that will seriously challenge any healthcare provider in the world. Despite a number of improvements in patient care, we are still unable to ameliorate or prevent the progression towards end-stage renal disease in the diabetic population. Hypertension and metabolic control appear to interact, resulting in the relentless decline in renal function observed in diabetic patients. Further understanding of the underlying mechanisms, and the development of new treatments against newly identified targets, is crucial for the prevention of this deadly microvascular diabetic complication. PMID- 19804274 TI - Diagnosis and management of Marfan syndrome. AB - Marfan syndrome is a disorder of the connective tissue that is inherited in an autosomal-dominant fashion and is caused by mutations in the gene coding for fibrillin-1, FBN1. Although complications of the syndrome may involve the eye, the lung and the skeleton, the high mortality of untreated cases results almost exclusively from cardiovascular complications, including aortic dissection and rupture. Recently, a series of experiments has begun to elucidate the complex molecular etiology of Marfan syndrome, and a number of new heritable syndromes with an associated risk for aortic complications, such as Loeys-Dietz syndrome types I and II, have been described. The multiorgan involvement of many of these syndromes requires multidisciplinary expert centers that can increase the average life expectancy of affected patients from only 32 years to over 60 years. The present article both reviews classical standards of managing cardiovascular manifestations and outlines significant advances in recent research with focus on their impact on future diagnostic and therapeutic options. PMID- 19804275 TI - Benefits of the Mediterranean diet on cardiovascular disease. PMID- 19804276 TI - The future of implantable hemodynamic monitors. PMID- 19804278 TI - Renin-angiotensin system genes and coronary artery calcification. AB - Evaluation of: Kretowski A, McFann K, Hokanson JE et al.: Polymorphisms of the renin-angiotensin system genes predict progression of subclinical coronary atherosclerosis. Diabetes 56, 863-871 (2007). The mechanisms giving rise to the increased risk of coronary artery disease (CAD) in patients with Type 1 diabetes compared with the general population are still incompletely understood. In the study considered here, coronary artery calcification (CAC), a marker of subclinical CAD, was measured over a mean 2.5-year period in approximately 500 subjects with Type 1 diabetes and 500 controls. Polymorphisms in the renin angiotensin system genes were found to be predictive of CAC progression in diabetic subjects, but only in the absence of treatment with angiotensin converting enzyme inhibitors or angiotensin-receptor blockers. This association could help in understanding the development of CAD in diabetic patients. However, because of the dangers of overinterpreting subgroup analyses and multiple testing, the findings require replication in a confirmatory study. PMID- 19804279 TI - Role of cathepsin C in elastase-induced mouse abdominal aortic aneurysms. AB - Evaluation of: Pagano MB, Bartoli MA, Ennis TL et al.: Critical role of dipeptidyl peptidase I in neutrophil recruitment during the development of experimental abdominal aortic aneurysms. Proc. Natl Acad. Sci. USA 104(8), 2855 2860 (2007). In this study, the authors used dipeptidyl peptidase I (DPPI) gene targeted mice and an aortic elastase perfusion-induced mouse abdominal aortic aneurysm (AAA) model to examine the role of DPPI, also termed cathepsin C, in the development of AAA. Mice lacking this protease are resistant to AAA formation. Interestingly, these authors found that DPPI activity controls neutrophil recruitment to the sites of inflammation, specifically AAA lesions in this case. By producing chemokine CXCL2, neutrophils in AAA lesions recruit additional neutrophils to the lesion sites where these cells utilize DPPI to activate neutrophil serine proteases, including neutrophil elastase, cathepsin G and proteinase 3, which may be further used to stimulate macrophage cytokine and chemokine production. In addition to DPPI-deficient mice, the authors also used antibodies against neutrophils (Gr-1) or CXCL2 receptor, CXCR2, to deplete neutrophils or to block the action of neutrophil chemokines to affirm their hypothesis. PMID- 19804280 TI - Management of atherosclerosis with antiallergic medicine: a lesson from the mouse model. AB - Evaluation of: Sun J, Sukhova GK, Wolters PJ et al.: Mast cells promote atherosclerosis by releasing pro-inflammatory cytokines. Nat. Med. 13, 719-724 (2007). Mast cells are important components in human allergic response and innate immunity. These cells have been implicated in the pathogenesis of atherosclerosis since the 1950s, and a series of studies have proposed their roles in the pathologic events critical to atherogenesis. Despite these studies and hypotheses, there is no evidence to suggest a direct participation of these allergic cells in atherosclerosis. Using mast cell-deficient mice and intravenous mast cell reconstitution technology with a conventional mouse atherosclerosis model, Dr Sun et al. revealed that mast cells contribute to atherogenesis by releasing proinflammatory cytokines, which are utilized to stimulate vascular cell-protease expression and further tissue remodeling. Mice that lack these cells are resistant to diet-induced atherosclerosis. These data suggest that stabilization of mast cells with antiallergic medicine may be utilized in controlling or preventing the initiation and progression of atherosclerosis in humans. PMID- 19804281 TI - Genomics, haplotypes and cardiovascular disease. AB - Cardiovascular disease has a complex genetic and environmental origin. Single gene mutations have been identified for a variety of disorders, including several forms of sudden cardiac death, atrial fibrillation, hypertrophic cardiomyopathy and coronary artery disease. The recent availability of haplotype data has further enabled genomic approaches to mapping genetic variants associated with the more common polygenic forms of cardiovascular disease. Genome-wide association studies have identified single nucleotide polymorphisms associated with coronary artery disease and are being applied to a variety of clinical problems such as in-stent restenosis. The combination of high-throughput genomic tools such as high density microarrays, genomic information such as sequence and haplotype data, and the careful clinical definition of phenotypes provides the framework for realizing the goals of personalized medicine. PMID- 19804282 TI - Role of the sarcomeric Z-disc in the pathogenesis of cardiomyopathy. AB - The Z-disc has traditionally been viewed as a structure required to maintain sarcomeric function and integrity. More recently, the sarcomeric Z-disc has also emerged as a nodal point in cardiomyocyte signaling and mechanotransduction. This notion is not only supported by several transgenic animal models, but also by the identification of mutations in various Z-disc proteins, resulting in dilated or hypertrophic cardiomyopathy in patients. This review will thus focus on the role of the sarcomeric Z-disc and its associated proteins in the pathogenesis of cardiomyopathy. PMID- 19804283 TI - Current and future treatment for pericarditis. AB - Acute pericarditis is a common disorder in several clinical settings, and may be the first manifestation of an underlying systemic disease. Disease management is often troublesome and concerns may arise regarding the fear of missing a specific etiology, the possible complications, particularly the evolution toward constrictive pericarditis, and the search for new drugs, following a presumed failure of current treatments. The response is often disproportionate, with extensive repeated diagnostic work-up, frequent readmissions and increasing use of new drugs with a doubtful risk-benefit ratio, often without an evidence-based approach. Frustration and management mistakes are common causes of a prolonged course of the disease and frequent consultations. A vicious circle is thereby maintained and is fostered by the relative lack of clinical trials and evidence based guidelines. The aim of this paper is to review more recent studies and trials in order to develop a more evidence-based management of the disease. PMID- 19804284 TI - Vascular progenitor cells and atherosclerosis. AB - Vascular regeneration occurs throughout life as a dynamic process. Millions of new endothelial cells are created with essentially the same number of cells undergoing programmed cell death or necrosis every day. As a result, the human vascular tree could be considered to essentially replace its entire endothelial population over a specified number of years. Within this network there is a compartment of vascular progenitor cells that appear to govern this homeostasis throughout life, continuously repopulating cells that die by apoptosis or necrosis. This delicate equilibrium appears to be disrupted in atherosclerotic disease processes as patients with known ischemic heart disease risk factors have been found to have lower numbers of circulating endothelial progenitor cells, which may tip the balance in favor of lesion formation, rather than repair. The aim of this article is to discuss the types of vascular progenitor cells and the mechanisms behind their mobilization, homing and differentiation into mature endothelial cells capable of vascular repair. PMID- 19804285 TI - Potential of adiponectin as a cardioprotective agent. AB - In this review, we focus on the role of adiponectin as a cardioprotective agent in several pathological heart conditions. Obesity is closely associated with Type 2 diabetes, hypertension and heart disease. Adiponectin is an adipose tissue derived hormone whose concentration is downregulated in subjects with obesity related diseases. Hypoadiponectinemia has been identified as an independent risk factor for Type 2 diabetes, coronary artery disease, acute coronary syndrome and hypertension. More recent experimental findings have shown that adiponectin directly affects signaling in cardiac myocytes and has beneficial effects on several pathological heart conditions, including cardiac hypertrophy and myocardial infarction. The favorable effects of adiponectin are associated with attenuated inflammatory response, decreased myocyte death, decreased hypertrophic response, maintained ischemia-induced angiogenesis and reduced interstitial fibrosis. Therefore, adiponectin could represent a molecular target for treating obesity-linked cardiac diseases. PMID- 19804286 TI - I(f) channels as a therapeutic target in heart disease. AB - In the normal heart, impulses are generated from the sinoatrial node. It is generally accepted that the pacemaker current, I(f), plays a major role in the spontaneous rhythmic activity. Recently, several electrophysiological and molecular data demonstrate that I(f) channels are present in embryonic and post natal ventricular myocytes and undergo a downregulation during maturation. Interestingly, the I(f) current is re-expressed in some pathological conditions such as cardiac hypertrophy and heart failure. In these conditions, the overexpression of f-channels is a consequence of electrophysiological remodeling and may represent an arrhythmogenic mechanism in heart failure, a condition associated with high risk for sudden cardiac death. For its physiological and pathophysiological role and the availability of selective f-channel blockers, I(f) may be a suitable therapeutic target in heart failure. PMID- 19804287 TI - Importance of screening to prevent heart attacks. PMID- 19804288 TI - Natriuretic peptide levels: what's on the horizon? PMID- 19804290 TI - Rolofylline (KW-3902): a new adenosine A1-receptor antagonist for acute congestive heart failure. AB - Rolofylline (KW-3902 or MK-7418) is an adenosine A1-receptor antagonist that exerts its effect by blocking adenosine-mediated constriction of the afferent glomerular arteriole. By blocking A1 receptors, rolofylline increases the glomerular blood flow and filtration and inhibits sodium reabsorption in the proximal tubule, thereby enhancing natriuresis and diuresis. Early phase clinical research data provided the proof-of-concept of an incremental effect beyond standard diuretic therapy. A large Phase III program is currently ongoing for the intravenous formulation of rolofylline in the treatment of acute heart failure. PMID- 19804291 TI - A vaccine against atherosclerosis: myth or reality? AB - Atherosclerosis is a chronic inflammatory disease that develops in the context of enhanced serum lipid levels. Nowadays, many studies focus on the modulation of inflammatory responses to reduce atherosclerosis. The most powerful strategy to achieve this is vaccination. In several immune diseases vaccination is shown to be very effective, resulting in a drastic decline in the incidence of the disease. But is vaccination also realistic in atherosclerosis? In this article, several approaches to vaccinate against atherosclerosis are described. Vaccination (based on protein or DNA) against bioactive molecules and disease related proteins successfully reduces experimental atherosclerosis. In addition, passive immunization with antibodies against atherosclerosis-specific antigens and tolerance induction, in which antigen-specific regulatory T cells are elicited, are described. In the near future, we expect an increased interest in vaccination against atherosclerosis and, maybe, the myth may become reality when the first clinical trials are performed. PMID- 19804292 TI - Gene-therapy delivery strategies in cardiology. AB - Clinical gene-therapy approaches in cardiology have not fulfilled their promise in randomized, controlled trials, so far, despite striking effects in preclinical models. Lack of clinical success appears not to be related to an unexpected low potency of the therapeutic factors itself in humans, but has rather been attributed to limitations of the vector systems used to transfer the DNA, as well as application modes of the vector itself. Therefore, novel delivery strategies are required with increased efficiency and increased specificity. Recent improvements of vectors using targeting approaches in addition to the development of novel application strategies for cardiac or vascular gene transfer will improve gene delivery in future clinical approaches. PMID- 19804293 TI - Heat-shock protein 60 and cardiovascular disease: a paradoxical role. AB - Heat-shock proteins (HSPs) are members of a highly conserved group of proteins that are induced in response to stress and injury. These proteins have protective properties, and can protect the heart from injury. HSP60 is found in the mitochondria and cytosol, and has essential intracellular functions including folding key proteins after their import into the mitochondria. In the cytosol, HSP60 binds to proapoptotic proteins, sequestering them. HSPs are highly conserved and, thus, are similar to bacterial proteins. Many individuals have antibodies to HSP60, possibly from prior infections. HSP60 can be found in the plasma membrane and in the serum in disease states. Serum HSP60 may be a marker for coronary artery disease. Once extracellular, HSP60 can cause cell injury. Thus, this protein has dichotomous functions for which the role in disease remains to be fully elucidated. PMID- 19804294 TI - Cardiovascular protection by angiotensin-converting enzyme 2: a new paradigm. AB - A novel angiotensin-converting enzyme (ACE) homolog, named ACE2, was recently described. ACE2 degrades Ang II, a peptide with vasoconstrictive and proliferative effects, to generate Ang-(1-7), which, acting through its receptor Mas, exerts vasodilatory and antiproliferative actions. In addition, ACE2 is a multifunctional enzyme and its actions on other vasoactive peptides can also contribute to its vasoactive effects. The discovery of ACE2 corroborates the establishment of two counter-regulatory arms within the renin-angiotensin system. The first arm is formed by the classical pathway involving the ACE-Ang II-AT(1) receptor axis, and the second arm is constituted by the ACE2-Ang-(1-7)-Mas receptor axis. Owing to its characteristics, the ACE2-Ang-(1-7)-Mas axis may represent new possibilities for developing novel therapeutic strategies for the treatment of hypertension and cardiovascular diseases. In this review, we will summarize the biochemical and pathophysiological aspects of ACE2 with particular focus on its role in the heart. PMID- 19804295 TI - Nitric oxide synthase inhibitors in cardiogenic shock: present and future. AB - Cardiogenic shock (CS) accompanying myocardial infarction carries a case fatality rate of 40-50%. Profound myocardial dysfunction is partially reversible, and possibly related to a state of inflammatory storm accompanied by nitric oxide (NO) overproduction. CS survivors enjoy satisfactory longevity and quality of life. The focus of this review is to describe the available data regarding NO synthase (NOS) inhibitors in CS. In view of supportive evidence from mammalian research (inducible-NOS-knockout mice are less susceptible to ischemic and reperfusion injury), therapies mitigating NO overproduction were tested in human CS subjects. Human randomized clinical trials project excellent safety but lack of efficacy. Although the Phase III, multicenter, prospective, randomized, double blind, placebo-controlled Study to Assess the Safety and Efficacy of Tilarginine Acetate (L-N(G)-monomethyl arginine citrate [L-NMMA]) in CS (TRIUMPH) trial demonstrated lack of clinical benefit of 5-h infusion of L-NMMA in CS, major design issues regarding the optimal timing, dosing, duration and NOS inhibitor need to be addressed prior to rendering this therapy ineffective. PMID- 19804296 TI - Future of cardiac resynchronization therapy. AB - Cardiac resynchronization therapy (CRT) has proven to be a beneficial treatment option in patients with severe drug refractory heart failure in the presence of electromechanical dyssynchrony. More recent trials have demonstrated mortality benefits associated with CRT, and even further reductions when combined with an internal cardiac defibrillator. Addressing the 20-30% cohort of patients who do not derive benefit from this novel therapy is a rapidly emerging area of research activity with encouraging results. Here we review the CRT trial evidence that forms the basis of patient-selection guidelines for device implantation and describe the present outstanding issues, alongside identifying future trends in CRT that appear promising. PMID- 19804298 TI - Role of personality in coronary heart disease: implications for behavioral cardiology. PMID- 19804297 TI - Pak1: steps towards understanding the regulatory mechanisms of pacemaker function of the heart. PMID- 19804300 TI - Does air pollution lead to atherosclerosis? PMID- 19804299 TI - Impacts of heart disease and depression on health-related quality of life. PMID- 19804301 TI - Rosiglitazone: a disappointing DREAM. AB - Dr Steven Nissen is a heart specialist and currently holds the position of chairman of cardiovascular medicine at the Cleveland Clinic, OH, USA. His work has involved the development of miniaturised ultrasound imaging devices that can be threaded into a patient's heart that allow measurement of the size and composition of plaques, which indicate early artery damage. The ability to characterize and measure the size of plaques provided a novel, effective method to evaluate the efficacy of anticholesterol medications, and for the past two decades Dr Nissen has been using these and other techniques to examine the efficacy of drugs. He has also developed a strong interest in drug safety. His work linked COX-2 inhibitors such as Celebrex and Vioxx (Merck, NJ, USA) with heart attacks, and prevented Merck's similar product, Arcoxia, from being approved. He also highlighted the serious heart attack risk associated with the experimental drug Pargluva and the drug was subsequently not approved by the US FDA. More recently, Dr Nissen's work has focused on the drug rosiglitazone, which was shown to have high cardiovascular risks and has since been given a FDA warning. Here, Dr Nissen discusses the publication of the rosiglitazone meta analysis and why he considers work in this area to be crucially important for patients. PMID- 19804302 TI - Mending broken hearts: the future of drug-based cardiac regeneration. AB - Coronary heart disease is a massive health problem worldwide. In the USA alone, 1.2 million people suffer a coronary attack every year, and approximately 40% of these individuals die as a result. As damaged cardiac muscle forms scar tissue and is unable to function properly, those that do survive the attack are often left severely ill. Consequently, the potential for a therapy that allows healthy cardiac muscle to proliferate at the site of injury is enormous. Dr Mark Keating is a leading researcher in this area of regenerative medicine and his group has recently identified a compound that aids in the process of cardiac regeneration. Although currently focussing on ophthalmic drug research at Novartis, Dr Keating still has a great interest in this area. Here he discusses the potential of drug based cardiac regeneration and the problems that must be overcome before this treatment can reach the clinic. PMID- 19804303 TI - Rimonabant: a novel approach for the treatment of obesity and cardiometabolic risk by blockade of the endocannabinoid system. AB - Obesity, the metabolic syndrome, diabetes and cardiometabolic risk are increasing at epidemic rates worldwide. Without interventions, the healthcare ramifications and costs of this epidemic will be astronomical. Current available treatment modalities have demonstrated limited effectiveness to deal with this metabolic epidemic. A novel metabolic pathway, the endocannabinoid system, plays a significant and direct role in appetite regulation, glucose homeostasis and lipid metabolism. Activation of the endocannabinoid system increases hunger and decreases satiety, and promotes insulin resistance and lipogenesis. Studies indicate that the endocannabinoid system is chronically activated in abdominal obesity and Type 2 diabetes. Additionally, blockade of the endocannabiniod receptor type-1 improves multiple cardiometabolic parameters and may represent a potential mechanism to combat obesity, metabolic syndrome, diabetes and other cardiometabolic risks. Rimonabant, a novel agent, blocks the endocannabinoid receptor type 1, and results in weight loss, decreases in abdominal adiposity, improvement in glucose and lipid homeostasis and decreases components of the metabolic syndrome. It is the first therapeutic agent that inhibits the endocannabiniod system and improves multiple cardiometabolic parameters. PMID- 19804304 TI - WATCHMAN left atrial appendage system for stroke prevention in atrial fibrillation: a percutaneous-device delivery approach. AB - Sick PB, Schuler G, Hauptmann KE et al.: Initial worldwide experience with the WATCHMAN left atrial appendage system for stroke prevention in atrial fibrillation. J. Am. Coll. Cardiol. 49(13), 1490-1495 (2007). Left atrium appendage (LAA) occlusion is a potential alternative to warfarin in patients with atrial fibrillation who have contraindications to anticoagulation. Currently, there are two devices specifically designed for LAA occlusion: the percutaneous LAA transcatheter occlusion (PLAATO System, ev3 Inc., MN, USA) and the WATCHMAN LAA system (Atritech Inc., MN, USA). Initial results from a study with the WATCHMAN device suggest that the LAA occlusion may reduce the long-term risk for stroke. Available data are still very limited and although occluding the LAA appears to be technically feasible, its long-term safety and ability to reduce stroke incidence remains to be proven. Only prospective randomized studies will be able to tell us if the LAA occlusion devices can be used as an alternative treatment strategy to the standard long-term anticoagulation. PMID- 19804305 TI - Antithrombotic dilemma. AB - Karjalainen PP, Porela P, Ylitalo A et al.: Safety and efficacy of combined antiplatelet-warfarin therapy after coronary stenting. Eur. Heart J. 28(6), 726 732 (2007). Chronic oral antithrombotic treatment is necessary in patients with mechanical heart valves and in the majority of patients with atrial fibrillation. Many of these patients also have ischemic heart disease. When these patients are required to undergo percutaneous coronary intervention with stenting, there is also an indication for treatment with aspirin and clopidogrel. However, triple therapy is known to augment the risk for bleeding complications. It is therefore unknown what the best antithrombotic treatment is, when considering both thrombotic (e.g., stent thrombosis) and bleeding complications. Unfortunately, no prospective (randomized) data are available to solve this issue. PMID- 19804306 TI - Bringing cardiac cell therapy with bone marrow stem cells to the clinic: where are we now? AB - Stem cell therapy is emerging as a potential therapeutic option for cell death related heart diseases. Preclinical as well as early-phase human studies have demonstrated the ability for cell therapy to augment perfusion and increase myocardial contractility. In addition, recent intermediate-size randomized trials suggested the potential of bone marrow stem cells to augment left ventricular recovery after a recent myocardial infarction. In general, the effects are modest and often similar despite differences in the study design, cell number or type. Therefore, a number of issues should be addressed before stem cell therapy will become standard clinical practice. They are related to the selection of the optimal cell type, standardization of the cell processing and release criteria. Other issues include timing of the cells injections and cell homing and retention. Further research is needed to understand the mechanisms underlying observed functional and beneficial effects including optimization of myocardial biological effects. Finally, despite overall enthusiasm, safety of the cardiac stem cell therapy should remain under scrutiny. The safety profile needs to be established in the large clinical trials and in a close interaction between translational and clinical research to address conceptual or procedural issues. PMID- 19804307 TI - Albuminuria: pathophysiology, epidemiology and clinical relevance of an emerging marker for cardiovascular disease. AB - Albuminuria has emerged from being a sign of early kidney disease in diabetes to an independent predictor of cardiovascular risk. Epidemiological studies suggest that microalbuminuria (30-300 mg/d) is present in 5-19% of the general population, in up to 23% of patients with hypertension and in up to 40% of patients with diabetes. Recent data suggest an even higher prevalence in certain patient populations. As it is associated with a variety of important cardiovascular risk factors, including prediabetes, dyslipidemia and the metabolic syndrome, detection of albumin in the urine represents an important diagnostic window for systemic micro- or macrovascular damage. Various studies have demonstrated the predictive value of all levels of albuminuria for future cardiovascular events in patients with diabetes, hypertension or overt cardiovascular disease, as well as in the general population. Annual screening for microalbuminuria is now recommended by international diabetes guidelines for patients with diabetes, and may be appropriate for nondiabetics with risk factors for cardiovascular disease. PMID- 19804308 TI - Potential of metabolic agents as adjunct therapies in heart failure. AB - Heart failure continues to have a significant morbidity and mortality rate despite several recent advances in treatment such as additional neurohumoral blockades and cardiac resynchronization therapy. There is emerging evidence that, irrespective of etiology, heart failure is associated with an energetic disorder and that this may contribute to the pathogenesis of the syndrome. Recently, a number of studies have suggested that some metabolic agents may have potential as adjunctive therapy in patients with heart failure. These agents cause a shift of myocardial-substrate utilization away from free fatty acids toward glucose. Free fatty acid utilization consumes more oxygen to generate an equivalent amount of energy compared with glucose. Some of these agents are also effective antianginals, presumably by reducing the myocardial oxygen requirement. In this review we will discuss some of the current issues and progresses relating to metabolic manipulation in heart failure. PMID- 19804309 TI - Role of apolipoprotein E genotype in coronary artery disease. AB - Recent gene-targeting technology has provided good animal models that provide insight into the pathology of complex diseases such as atherosclerosis. The apolipoprotein E gene polymorphism is one of the most extensively studied in cardiovascular medicine. The scope of the present review is to briefly outline the biochemical characteristics and the genetic variation of apolipoprotein E. Apolipoprotein E is best known for its role in modulating lipoprotein metabolism as a ligand for cellular receptors. Other functions unrelated to lipid transport are becoming known, including reverse cholesterol transport, immunoregulation and modulation of cell growth. This review will examine recent work that addresses how apolipoprotein E participates in atherosclerosis. Genotypic variation of apolipoprotein E has been associated with certain phenotypes regarding vascular disease, such as the presence of atherosclerosis and coronary heart disease outcomes. This article will also review evidence regarding the association between apolipoprotein E gene polymorphisms and coronary artery disease based upon experimental and clinical studies. PMID- 19804310 TI - Update on the role of the atherogenic lipoprotein phenotype in cardiovascular prevention. AB - Higher plasma triglyceride levels and decreased HDL-cholesterol concentrations are usually accompanied by the presence of small, dense LDL in the so-called lipid triad or 'atherogenic lipoprotein phenotype'. This phenotype is highly atherogenic and its prevalence may suggest an even higher overall burden of atherosclerotic disease as compared with that associated with hypercholesterolemia. As stated by the National Cholesterol Education Program Adult Treatment Panel III, there is evidence suggesting each component of this lipid triad is individually atherogenic. However, the relative contribution of each component cannot be easily determined. Therefore, it has been suggested to consider the atherogenic lipoprotein phenotype as a whole as a risk factor. This is supported by data from epidemiological studies considering high-risk populations, which showed that the contribution to cardiovascular risk of each individual component cannot be dissected from the sum of all factors. We recently investigated the prevalence of the atherogenic lipoprotein phenotype in different categories of patients at higher cardiovascular risk: with coronary and noncoronary forms of atherosclerosis or metabolic diseases, including Type 2 diabetes, polycystic ovary syndrome and growth hormone deficiency. Subjects with higher triglyceride levels, decreased HDL-cholesterol concentrations and increased levels of small, dense LDL (i.e., subjects with the atherogenic lipoprotein phenotype) are common in coronary and noncoronary forms of atherosclerosis. In the future, it may be possible to measure the presence of small, dense LDL to identify subgroups at higher cardiovascular risk. PMID- 19804311 TI - Coronary heart disease and pregnancy. AB - The prevalence of coronary artery disease in female patients is increasing due to changing lifestyle patterns including cigarette smoking, diabetes and stress. Since women are delaying childbearing until older age, acute coronary syndrome will more frequently occur during pregnancy. Although rare, acute coronary syndrome during pregnancy often has devastating consequences. It is associated with increased maternal and neonatal mortality and morbidity compared with the nonpregnant situation. Furthermore, it constitutes an important problem for the patient and the treating physician, because the selection of diagnostic and therapeutic approaches is greatly influenced not only by maternal, but also by fetal safety. PMID- 19804313 TI - Strategies for the regulation of intracellular calcium in ischemic heart disease. PMID- 19804314 TI - Chemokines in myocardial infarction: translating basic research into clinical medicine. PMID- 19804316 TI - Olmesartan for the treatment of arterial hypertension. AB - Angiotensin-receptor blockers (ARBs) are an important class of agents used for the treatment of arterial hypertension. Olmesartan medoxomil, the seventh latest ARB approved by the US FDA, is an oral, once-daily, AT(1)-receptor selective ARB with high receptor affinity. Pharmacologically, it acts as a competitive and insurmountable Ang II antagonist with linear pharmacokinetics and without cytochrome P450 interaction. The drug is licensed for the treatment of arterial hypertension alone or in combination with other antihypertensive agents. Olmesartan has demonstrated its dose-dependent inhibitory effect on Ang I-induced blood pressure responses between 10 and 80 mg in Phase II studies. These results, confirmed in an international clinical trial programme covering over 3000 hypertensive patients in numerous studies, demonstrated rapid blood pressure lowering effects within 1 week. A daily oral dose of 20 mg olmesartan is considered to be the optimal dose. In clinical trials and postmarketing studies, olmesartan has been shown to be safe and well tolerated with an adverse event profile similar to the placebo. Active comparative studies demonstrated either similar or superior efficacy of olmesartan compared with other ARBs, angiotensin converting enzymes inhibitors, beta-blockers or calcium-channel blockers. Besides its antihypertensive efficacy, olmesartan was shown in clinical trials to reduce vascular microinflammation, decrease intrarenal vascular resistance, significantly reduce vascular remodeling of small resistance arteries and exert antiatherosclerotic effects by significantly reducing the volume of large atherosclerotic plaques. PMID- 19804317 TI - New concepts in transcatheter closure of paravalvular leaks. AB - New catheter techniques and a large variety of endovascular devices permit transcatheter treatment - not only of common atrial or ventricular septal defects - but also of rare intracardiac malformations such as postmyocardial infarction ventricular septal defects, sinus valsalva aneurysm or paravalvular leak. Owing to the variable anatomical nature of paravalvular leaks, closure may be one of the most challenging interventional procedures in the field of structural heart disease. Current literature documents ambiguous results and endorses the limitations of this procedure. In most cases these limitations are technical. Further efforts in the development of defect-specific devices and techniques may result in an improved patient outcome and extended patient selection. PMID- 19804318 TI - Genetics of congenital long QT syndrome and Brugada syndrome. AB - The inherited cardiac arrhythmias including congenital and acquired long QT syndrome (LQTS), Brugada syndrome, progressive cardiac conduction defect, catecholaminergic polymorphic ventricular tachycardia, arrhythmogenic right ventricular cardiomyopathy, familial atrial fibrillation, familial sick sinus syndrome and short QT syndrome, are linked to mutations in genes encoding for ion channels or other membrane components. Eleven forms of congenital LQTS have been identified and these are caused by mutations in genes of the potassium, sodium and calcium channels or membrane adapter. Genotype-phenotype correlations have been rigorously investigated, especially in the LQT1, LQT2 and LQT3 forms, which constitute more than 90% of genotyped patients. On the other hand, causative mutations were identified much less in patients with Brugada syndrome, therefore data on genotype-phenotype relationships are limited. PMID- 19804319 TI - Treatment of ST-elevation myocardial infarction. AB - In patients with ST-segment elevation myocardial infarction (STEMI), timely and adequate treatment may improve the prognosis dramatically. Restoration of the infarct vessel patency is one of the cornerstones of initial treatment. Compared with fibrinolytic therapy, primary percutaneous coronary intervention (PCI) results in improved short- and long-term survival, a lower incidence of recurrent infarction and a better left ventricular function. Although (drug-eluting) stents may reduce restenosis, effects on mortality are less clear. Administration of glycoprotein IIb/IIIa antagonists may further reduce periprocedural coronary complications, but bivalirudin may offer similar effects with less bleeding. beta adrenergic blockers, angiotensin-converting-enzyme inhibitors and statins should be initiated in all patients with STEMI, although cautious use of beta-blockers is advised in patients at risk of cardiac shock. Patients with diabetes should receive optimal glucose control. High-risk patients, particularly those with a low ejection fraction, should receive an implantable cardioverter defibrillator after 30 days, although it is not clear whether patients who have received primary PCI also benefit, particularly if they have no signs of heart failure. PMID- 19804320 TI - Drawbacks to stem cell therapy in cardiovascular diseases. AB - Stem cells seem to have unlimited potential for repairing injured tissues derived from cardiovascular diseases. Much as the initial euphoria over preclinical models has ushered in some skepticism, several reports have advised caution against over exuberance, as cellular therapy has both theoretical and reported safety concerns. Embryonic stem cells, skeletal myoblasts, bone marrow-derived stem cells and mesenchymal stem cells are current candidates for cell therapy in end-stage cardiovascular diseases. However, before large-scale clinical trials can take place, a few safety concerns have to be clarified, such as atherogenesis, postangioplasty or stenting restenosis, tumorigenesis, stem cell metastasis, stem cell-mobilized cytokine-related complications and arrhythmogenesis. In this review, potential ways to overcome these issues are discussed, including medical and gene manipulations, dedicated cell-purification techniques, antiarrhythmic cell therapy design, and new concepts such as using natural constructs. With these safety issues under control, stem cell therapy still has a promising future in the next decade. PMID- 19804321 TI - Therapeutic potential of novel modulators of neovascularization. AB - Neovacularization is an important biological process whereby new blood vessels develop in both health and disease. During development, blood vessels are formed from mesodermal cells in a process called vasculogenesis. The vascular network then expands by the sprouting of new vessel networks from pre-established vessels in a process known as angiogenesis. However, in adult life, undesirable neovascularization is associated with tumor development and a growing list of 'angiogenesis-dependent' diseases, including cardiovascular complications. Furthermore, diseases characterized by ischemia-induced tissue damage cause a neovascularization response to facilitate tissue repair. Recent research has identified novel molecular and cellular mediators of neovascularization that, in adult life, recapitulate angiogenic processes observed during embryonic development. The discovery of vascular progenitor cells and new molecules that display selective functions in modulating endothelial cell fate, migration and patterning, vessel morphogenesis and the amplification of angiogenic signaling by regulating the master signal VEGF, opens the door to new clinical strategies that target angiogenesis-dependent diseases or that can promote therapeutic neovascularization. PMID- 19804324 TI - Calcium-channel blockers and cardiac arrhythmias within the elderly population. PMID- 19804322 TI - gp130-mediated pathway and heart failure. AB - Binding of ligands to gp130 activates at least three different downstream signaling pathways: the signal transducer and activator of transcription (STAT), the Src-homology tyrosine phosphatase 2-ras-MAPK and the PI3K/Akt pathways. Cardiac-specific disruption of gp130 was shown to result in heart failure in response to mechano-stress accompanied by an increase in apoptosis of cardiac myocytes. Inactivation of STAT3 resulting from the loss of gp130 may be a key event in the transition from cardiac hypertrophy to heart failure. Proper vascular growth would be essential for normal cardiac development and the remodeling process. In addition to various factors, such as bcl-xL, inducible nitric oxide synthase and reactive oxygen species-scavenging proteins, VEGF has also been identified as a target gene of STAT3 and together can promote cardiac myocyte survival by preventing apoptosis and restoration of energy deprivation. In this regard, the gp130-receptor system and its main downstream mediator, STAT3, play a key role in the prevention of heart failure. In this review, current knowledge of the IL-6 family of cytokines relating to human cardiac disease is summarized, in addition to the potential role of gp130-mediated signaling systems in various models of experimental heart failure. PMID- 19804325 TI - Prevention strategies for hypertension: who should be targeted? PMID- 19804326 TI - HYVET indicates stroke and all-cause mortality benefit for antihypertensive treatment in the very elderly. PMID- 19804327 TI - Perindopril: beyond lowering blood pressure. AB - Perindopril, a prodrug ester of perindoprilat, is an angiotensin-converting enzyme inhibitor that lowers angiotensin II and potentiates bradykinin. This agent has proven efficacy in a wide range of cardiovascular diseases. Its efficacy, safety and tolerability are well established in the treatment of hypertension. Beyond pharmacodynamic effects shown in lowering blood pressure, perindopril was also involved in the improvement of endothelial function and the normalization of vascular and cardiac structure and function. Large morbidity mortality trials, such as the European Trial on Reduction of Cardiac Events with Perindopril in Patients with Stable Coronary Artery Disease (EUROPA), Perindopril Protection Against Recurrent Stroke Study (PROGRESS) and Anglo-Scandinavian Cardiac Outcomes Trial-Blood Pressure Lowering Arm (ASCOT-BPLA), have shown that treatment with perindopril reduces and prevents cardiovascular disease in a large range of patients with vascular diseases (including stable coronary disease), whether or not they are hypertensive. PMID- 19804328 TI - Aging and the brain renin-angiotensin system: relevance to age-related decline in cardiac function. AB - This article discusses evidence that impairments in control of autonomic outflow mediated by the brain renin-angiotensin system (RAS) contribute to the decline in baroreceptor reflex function and the development of insulin resistance that accompany hypertension and excess salt intake, especially during aging. Imbalances in the regulation of the sympathetic and parasympathetic limbs of the autonomic nervous system observed in older subjects underlie changes in heart rate variability and play a role in the regulation of overall cardiac function. Age-related alterations in autonomic nervous system function may also explain the age-associated alterations in metabolism. Reduced heart-rate variability is linked to increased mortality in patients with cardiovascular disorders and, coupled with information that is known about local changes in the cardiac and brain RAS during aging, the evidence reveals potential mechanisms for the protective effects of systemic blockade of the RAS against age-related changes that impact the heart. PMID- 19804329 TI - What modulates the aortic valve interstitial cell phenotype? AB - Age-related, degenerative aortic valve disease is of great concern for the current aging population, and the only current treatment option is replacement, necessitating open-chest surgery. While the initial disease mechanism may involve the endothelial cells that sheath the valve leaflets, there is strong evidence supporting the notion that it is in fact a malfunction of the robust interstitial cells within the leaflet tissue that perpetuate the disease through a presently unclear sequence of events. In this article, recent data are discussed regarding mechanobiology of the interstitial cell, with particular focus on modulation of the phenotype and subsequent biosynthetic function. Moreover, speculation of potential mechanisms that may ultimately lead to degenerative aortic valve disease are discussed. PMID- 19804330 TI - Real-time imaging in left atrial mapping and ablation. AB - The catheter-based ablation of atrial fibrillation has been transformed greatly by the introduction of new technologies and techniques. This article describes the major advancements in real-time navigation systems, including both 3D mapping systems and 2D echocardiography. The relative strengths and weakness of these systems and their accuracy on clinical outcome is also discussed. Finally, we explore current and emerging MRI technologies that will allow the assessment of disease progression and enable procedural planning. PMID- 19804331 TI - Thrombosis after stent implantation: how much of a problem is there? AB - Drug-eluting stents reduce the occurrence of in-stent restenosis and the need for subsequent target vessel revascularization when compared with bare-metal stents. However, drug-eluting stents may be associated with delayed (or absent) endothelialization, localized hypersensitivity reactions and late stent thrombosis. Stent thrombosis is a catastrophic event, resulting in life threatening complications. Although larger, adequately powered, randomized trials are needed to fully assess the net clinical effects of drug-eluting stents compared with bare-metal stents, the evidence, thus far, appears to suggest that the net clinical benefit of drug-eluting stents may outweigh their risks. Premature discontinuation of antiplatelet therapy is the most important predictor of stent thrombosis; therefore, patients who are candidates for implantation of drug-eluting stents should be screened for their ability to receive and tolerate uninterrupted antiplatelet therapy longer than is necessary with bare-metal stents. PMID- 19804332 TI - Role of noninvasive imaging using computed tomography for detection and quantification of coronary atherosclerosis. AB - Clinical cardiac computed tomography (CT) began with electron-beam CT in the early 1980s and continues now with multidetector CT in the 21st Century. The major applications of noncontrast cardiac CT are currently for the quantification of coronary artery calcium - a reliable and repeatable means to estimate atherosclerotic plaque burden. The major applications of contrast-enhanced CT (CT angiography) is for a more detailed estimation of total plaque burden by qualitatively defining noncalcified and complex plaque as well as ruling out obstructive coronary artery disease. Both of these applications are discussed and comments are made from the author regarding clinical applications based upon reviewing the published literature and through personal experience. PMID- 19804333 TI - Stem cell therapy for heart failure: the science and current progress. AB - Cell therapy, particularly with stem cells, has created great interest as a solution to the fact that there are limited treatments for postischemic heart disease and none that can regenerate damaged heart cells to strengthen cardiac performance. From the first efforts with myoblasts to recent clinical trials with bone marrow-derived stem cells, early reports of cell therapy suggest improvement in cardiac performance as well as other clinical end points. Based on these exciting but tentative results, other stem cell types are being explored for their particular advantages as a source of adult stem cells. Autologous adipose derived stem cells are multilinear and can be obtained relatively easily in large quantities from patients; cardiac-derived stem cells are highly appropriate for engraftment in their natural niche, the heart. Human umbilical cord blood cells are potentially forever young and allogenic adult mesenchymal stem cells appear not to evoke the graft versus host reaction. Human embryonic stem cells are effective and can be scaled up for supply purposes. The recent discovery of induced pluripotentcy in human adult stem cells, with only three transcription factor genes, opens a whole new approach to making autologous human pluripotent stem cells from skin or other available tissues. Despite the excitement, stem cells may have to be genetically modified with heme oxygenase, Akt or other genes to survive transplantation in a hypoxic environment. Homing factors and hormones secreted from transplanted stem cells may be more important than cells if they provide the necessary stimulus to trigger cardiac regrowth to replace scar tissue. As we await results from larger and more prolonged clinical trials, the science of stem cell therapy in cardiac disease keeps progressing. PMID- 19804334 TI - Future potential of echocardiography in heart failure. AB - Echocardiography represents a convenient, portable and noninvasive method to provide important anatomic and physiologic information to inform the management of heart failure patients. Traditional echo assessments include diagnostic, etiologic and prognostic data from ventricular size, geometry and performance. Newer echocardiographic techniques are receiving greater utilization, however, and promise to further enhance diagnostic abilities in heart failure. This article reviews traditional anatomic assessments, echo-based cardiac hemodynamics, 3D echocardiography, quantification of myocardial tissue mechanics and hand-carried echocardiography. These developments in echocardiography underlie future trends toward echo objectivity, improved imaging of patients with poor acoustic windows, miniaturization and simplicity in focused exams and the expanded application of old and new techniques. PMID- 19804335 TI - New techniques of mapping and ablation for tachyarrhythmias in children. AB - Advancement in mapping and ablation technologies in the past decade has made arrhythmia treatment in children safer and more effective. Electroanatomical mapping systems with features of nonfluoroscopic navigation, 3D-geometry construction and color-coded arrhythmia mapping have been extensively used to map and ablate tachycardias in children. In addition, a variety of cooled radiofrequency ablation systems have been used in selected patients to create larger and deeper lesions. Cryoenergy ablation, with its reversible nature of tissue injury during the cooling phase, is emerging as the energy of choice for ablating perinodal arrhythmias to minimize the risk of inadvertent conduction block. Other evolving ablation systems, including cryoballoon, high-intensity focused ultrasound balloon and a hybrid catheter capable of delivering both radiofrequency and cryoenergy, are also under investigation. Finally, a remote navigation system that minimizes radiation exposure and will enable physicians of all skill levels to reach difficult target sites in children is on the horizon. PMID- 19804337 TI - Assessment of myocardial viability: an effective gatekeeper for coronary revascularization? PMID- 19804338 TI - Carotid artery stenting: what do recent trials and registries tell us? PMID- 19804340 TI - Bosentan for pulmonary hypertension and other pulmonary diseases: emerging evidence. AB - Endothelin-1 is a potent vasoconstrictor and mitogen that is primarily synthesized and released from vascular endothelial cells. Bosentan is a dual endothelin-receptor antagonist that initially received approval for treatment of WHO group I pulmonary arterial hypertension (PAH) for patients in functional classes III and IV. Analysis of a study conducted in functional class II patients (Endothelin Antagonist Trial in Mildly Symptomatic PAH Patients [EARLY] trial) suggest its efficacy for these less symptomatic patients. In addition, bosentan has demonstrated efficacy in patients with congenital heart disease and Eisenmengers syndrome with right to left shunting and in HIV-related PAH. Studies of bosentan in inoperable or residual chronic thromboembolic pulmonary hypertension suggest possible efficacy. Bosentan appears promising in patients with idiopathic pulmonary fibrosis who do not have pulmonary hypertension. Combinations of bosentan with other PAH therapies such as iloprost and sildenafil may have incremental benefit over monotherapy. PMID- 19804341 TI - Pacemaker and defibrillator therapy in pediatrics and congenital heart disease. AB - Pacemakers and defibrillators have a growing use in pediatrics and in patients with congenital heart disease, but they present unique problems and implications for their implantation and follow-up. Congenital and surgically acquired rhythm disturbances are common, but the efficacy of device therapy is not well established in these patient groups. The diversity and complexity of pediatric patients and congenital heart disease make device management a highly individualized art. There are technical issues related to device implantation that have necessitated novel approaches to using leads and device that were not designed with children specifically in mind. The current guidelines and indications for implantable device therapy for children and congenital heart disease are reviewed, as well as some of the specific limitations and problems encountered. PMID- 19804342 TI - Mesenchymal stromal cell and mononuclear cell therapy in heart disease. AB - Despite progress in percutaneous coronary intervention, bypass surgery and drug therapy, rates of mortality and morbidity after acute coronary syndrome are high due to ventricular remodeling and heart failure. Mesenchymal stromal cells (MSCs) from adult bone marrow or adipose tissue are considered potential candidates for therapeutic regenerative treatment in cardiovascular disease. Recent animal studies have demonstrated that MSCs can induce neovascularization and improve myocardial function in postinfarction myocardial ischemic hearts. This review will focus on the present preclinical and clinical knowledge about the use of mononuclear cells and MSCs for cardiac regenerative medicine, the source of MSCs for clinical use and problems to consider when conducting clinical MSC therapy. PMID- 19804343 TI - Serum amyloid A as a marker and mediator of acute coronary syndromes. AB - Inflammation promotes acute coronary syndromes and ensuing clinical complications. An emerging downstream marker of inflammation is serum amyloid A (SAA). Elevated plasma SAA levels predict increased cardiovascular risk and portend worse prognosis in patients with acute coronary artery disease (CAD). The pathophysiological role of SAA remains enigmatic. SAA plays a role in host defense, but it might also be atherogenic. SAA affects cholesterol transport, contributes to endothelial dysfunction, promotes thrombosis, evokes recruitment of inflammatory cells, activates neutrophils and suppresses neutrophil apoptosis, key events underlying acute coronary syndromes. These results provide a potential link between SAA and CAD and suggest that reducing SAA levels and/or opposing the actions of SAA may have beneficial effects in patients with acute CAD. PMID- 19804344 TI - Cyclic nucleotide-gated channels: a familiar channel family with a new function? AB - The cyclic nucleotide-gated (CNG) channel is a family of nonselective cation channels that open in response to an elevated cyclic nucleotide level. Cyclic nucleotides, particularly cAMP and cGMP, govern a great diversity of cellular functions. While the pivotal roles of CNG channels in the visual and olfactory systems have been well established in the past decade, relatively few studies were performed regarding the functional roles of CNG channels in non-neuronal systems. Cyclic nucleotides and Ca2+ are key signaling molecules in cardiovascular systems. Given that CNG channels are expressed in vascular tissues, several recent studies have explored the possible functional role of CNG channels in cardiovascular systems. This article intends to summarize some recent developments regarding the expression and functional role of CNG channels in the cardiovascular system. PMID- 19804345 TI - Challenges in advanced chronic heart failure: drug therapy. AB - Despite advances in both drug and device treatment of chronic heart failure (CHF) over the last 20 years, many patients still progress to a stage of advanced CHF, characterized by increasing symptoms and declining functional status. Future drug management of such patients presents many challenges. This review focuses on the issue of optimizing standard medical therapy in advanced CHF, the treatment of diuretic resistance and hyponatremia. As well as prescribing drugs in this phase of the disease, the system of care used to deliver therapy is crucial. On its own, multiprofessional heart failure care can improve outcomes for these patients. Finally, this review also addresses the drugs and model of care used to deliver palliative care in the end stage of advanced CHF. PMID- 19804346 TI - Catheter ablation of ventricular tachycardia guided by noncontact mapping. AB - Catheter ablation of untolerated and unstable ventricular tachycardia may not be performed using a conventional activation mapping tecnique. The noncontact mapping system enables reconstruction of the spreading of activation wave through a virtually generated ventricular chamber, even from a single tachycardia beat, and was introduced as a tool to guide mapping and ablation of untolerated or unsustained ventricular arrhythmias. The reduced accuracy in the setting of enlarged ventricles is recognized as the main limitation of this tecnique. While noncontract mapping appears to be especially suitable in guiding the ablation of unsustained idiopathic ventricular arrhythmias, it can also be successfully used as a guide to perform ablation of untolerated re-entry-related ventricular tachycardias during sinus rhythm. PMID- 19804348 TI - Regenerative medicine with endometrial regenerative cells for critical ischemia: limb salvage from the cradle of life? PMID- 19804350 TI - Vernakalant (RSD1235) in the management of atrial fibrillation: a review of pharmacological properties, clinical efficacy and safety. AB - Vernakalant (RSD1235) is a novel antiarrhythmic agent for conversion of rapid onset atrial fibrillation (AF). It is an atria-selective multichannel ion blocker (blocks I(Kur), I(Na), I(Ca, L), I(to) and I(Kr)), with a small effect on ventricular repolarization. In clinical Phase II and III studies, vernakalant was moderately (approximately 50%) effective in converting AF of short duration (< 7 days), and effective (approximately 70-80%) in converting AF of less than 72 h, but was not effective in converting long duration AF (>7 days) or atrial flutter. Vernakalant seems to have only a small proarrhythmic effect, with no reported cases of torsades de pointes in direct relation to vernakalant administration in Phase II and III studies. Overall, there are few reported serious adverse events. PMID- 19804351 TI - Noninvasive techniques to assess myocardial ischemia in hypertensive patients. AB - Hypertensive patients are more affected by coronary artery disease (CAD) than normotensive patients. Currently, established techniques are able to diagnose myocardial ischemia/CAD in hypertensive patients with suspected CAD. An irrevocable role remains for exercise electrocardiogram (ECG) owing to the many parameters evaluated under physiologic conditions. However, the suboptimal specificity of a positive exercise ECG demands further examination of hypertensive patients with either myocardial single-photon emission computed tomography (SPECT) or stress echocardiography, both more specific than exercise ECG in diagnosing CAD. The high specificity of imaging techniques also makes them highly predictive of cardiac events. Additional techniques to help diagnose CAD are available. Tissue Doppler imaging, strain and other echo-derived techniques may add quantitative elements to recognize CAD in hypertensive hearts. The accuracy of MRI is improving to study myocardial function and perfusion. Positron emission tomography and multislice computed tomography, also in-built with fusion scanners, are playing roles in combining coronary angiography and myocardial function/ischemia assessment. However, no significant application of these additional techniques is available for hypertensive patients. Epicardial CAD assessed by coronary angiography remains the gold standard to decide for revascularization procedures. The presence of microcirculatory dysfunction, a symptom typical of hypertensive hearts, is opening up new areas of noninvasive diagnostic techniques for the detection of coronary flow reserve (CFR) and related myocardial ischemia. The quantification of CFR may render this parameter pivotal to deciding the need for revascularization procedures of intermediate coronary stenosis and it may become an additional gold standard in evaluating coronary vessels. Moreover, even with normal epicardial coronary arteries, microcirculation dysfunction bears prognostic stratification capabilities for hypertensive patients and it may become a promising therapeutic target in the near future. PMID- 19804352 TI - Novel methods to assess heart valve disease. AB - Optimal timing of surgery for heart valve disease relies on the accurate assessment of symptoms, lesion severity, cardiac function and the risks of disease progression. Recent studies suggest potential roles for new echocardiographic techniques, including tissue Doppler and strain imaging at rest or after exercise stress, cardiac magnetic resonance imaging and biomarkers such as B-type natriuretic peptide. These techniques may identify patients at higher risk of symptomatic deterioration or adverse clinical events, and improve the cost-effectiveness and reliability of follow-up. PMID- 19804353 TI - Natriuretic peptides and their evolving clinical applications. AB - Natriuretic peptides (NP) are quantitative plasma biomarkers of heart failure, which are widely used in clinical practice in many countries. NP levels are accurate in the diagnosis of heart failure in patients presenting with dyspnea. The use of NP improves patient management and reduces total treatment costs in patients with dyspnea. As NP levels quantify disease severity in patients with established heart failure, NP levels are powerful predictors of outcome in predicting death and rehospitalization. NP-guided therapy may improve morbidity in patients with chronic heart failure. Although NP levels also risk-stratify patients with many other conditions such as stable or unstable coronary artery disease, pulmonary embolism and community-acquired pneumonia, there is insufficient evidence on how patient outcome could be altered in patients identified as high risk. PMID- 19804354 TI - Colchicine for the treatment of pericarditis. AB - Colchicine has been effectively used in the treatment of several inflammatory conditions, such as gouty attacks, serositis related to familial Mediterranean fever, Behcet syndrome and more recently, in acute and recurrent pericarditis. Colchicine concentrates in white blood cells, particularly polymorphonuclear cells, inhibiting tubulin polymerization, thus interfering with migration and phagocytosis, and reducing the inflammatory cycle. Although the exact number of responders is unknown, the drug has been successfully used for the treatment and prevention of recurrences and to taper corticosteroids in patients with recurrent pericarditis in several retrospective studies and an open-label, randomized trial, where the recurrence rate was halved in the treatment arm. Less evidence supports the use of the drug for the treatment of acute pericarditis, where colchicine remains optional and requires further multicenter confirmatory studies. At present, colchicine has been recommended by the 2004 European guidelines on the management of pericardial diseases for acute (class IIa) and recurrent pericarditis (class I), but its use is still unlabeled and informed consent is required for prescription. A careful monitoring of possible contraindications, drug interactions and side effects is necessary. The aim of this paper is to review the evidence that supports the use of the drug in acute and recurrent pericarditis, as well as dosing and precautions for clinical use. PMID- 19804355 TI - Minimally invasive mitral valve surgery: evolution, techniques and outcomes. AB - Surgical treatment of the mitral valve has evolved considerably since the 1920s. During the mid-1990s new approaches through small incisions have been proposed to access the mitral valve. We herein describe the evolution of mitral surgery, the development of minimally invasive mitral techniques and compare outcomes to a standard sternotomy approach. PMID- 19804356 TI - How to avoid complications associated with carotid angioplasty and stenting. AB - Carotid angioplasty and stenting (CAS) has been proposed as an alternative to surgery and is now performed more frequently and is well accepted, at least for high surgical risk patients. However, complications and particularly embolic strokes, even with a meticulous technique, can occur at any step of the procedure. Silent embolism is detected after CAS and may be a problem that needs to be discussed. To avoid and reduce these complications associated with CAS, it is important to have good indications dependent upon on good patient and lesions selection, as well as correct technique. New parameters have been proposed, particularly for asymptomatic lesions. The authors consider that embolic protection devices (EPDs) are mandatory for CAS, and new techniques will be presented. The choice of the EPD depends on the clinical status of the patient, the lesion morphology and characteristics, and the anatomy of the artery. All stents are not equivalent and so a good choice of the stent is necessary to avoid and reduce the complications associated with CAS, and experienced operators are also needed. With all these considerations, CAS can now be performed with acceptable outcomes and in certain population the results are comparable or superior to surgery. The results of ongoing randomized trials are awaited. PMID- 19804357 TI - Past, current and future concepts in atherosclerotic biobanking. AB - Knowledge on the pathogenesis of atherosclerotic disease mainly originates from observations made by pathologists who systemically collected vascular tissue. Biobanking of human tissues has become a professionalized joint effort, including the expertise of epidemiologists, pathologists and biologists. Mostly, biobanks are used for cross-sectional studies and the obtained specimens often represent later stages of disease. Technical improvements in high-throughput genetic and proteomic screening lead to important new insights into pathophysiological processes. Atherosclerotic disease progression is a major killer in the Western society. Pathological biobanking studies revealed insights in plaque progression and destabilization. However, atherosclerotic disease is a typical example where insights into progression of the disease may be hampered by the cross-sectional design of the biobanks. This article will focus on the accomplishments that have been made through biobanking of atherosclerotic tissues in the past and the present. In addition, future potentials of atherosclerotic plaque biobanks will be discussed. PMID- 19804359 TI - EFEMP1 binds the EGF receptor and activates MAPK and Akt pathways in pancreatic carcinoma cells. AB - The EGF-related protein EFEMP1 (EGF-containing fibulin-like extracellular matrix protein 1) has been shown to promote tumor growth in human adenocarcinoma. To understand the mechanism of this action, the signal transduction activated upon treatment with this protein has been investigated. We show that EFEMP1 binds EGF receptor (EGFR) in a competitive manner relative to epidermal growth factor (EGF), implicating that EFEMP1 and EGF share the same or adjacent binding sites on the EGFR. Treatment of pancreatic carcinoma cells with purified EFEMP1 activates autophosphorylation of EGFR at the positions Tyr-992 and Tyr-1068, but not at the position Tyr-1048. This signal is further transduced to phosphorylation of Akt at position Thr-308 and p44/p42 MAPK (mitogen-activated protein kinase) at positions Thr-202 and Tyr-204. These downstream phosphorylation events can be inhibited by treatment with the EGFR kinase inhibitor PD 153035. The observed signal transduction upon treatment with EFEMP1 can contribute to the enhancement of tumor growth shown in pancreatic carcinoma cells overexpressing EFEMP1. PMID- 19804360 TI - Binding and activation of the human plasma kinin-forming system on the cell walls of Candida albicans and Candida tropicalis. AB - Bacterial infections often upregulate the plasma kinin-forming cascade of the host (the 'contact system') which is triggered by adsorption of high molecular mass kininogen (HK), coagulation factor XII (FXII) and prekallikrein (pHPK) on the host or pathogen cell surfaces. A possible activation of the contact system upon infection of the human host by major fungal pathogens of Candida species has not been extensively explored until a recent report of tight binding of HK to the cell walls of these fungi. In the current study, the adsorption of the other contact system components to the cell surfaces of Candida albicans and Candida tropicalis was characterized. FXII was found to be tightly bound by Candida germ tube forms, to a level 5-fold higher than that for HK. In contrast, pHPK bound poorly but its additional amounts could dock to the cell wall through the surface bound HK. It was also shown that within the complex of these proteins assembled on the cell walls of fungal hyphae, pHPK could be activated by FXIIa and the active HPK effectively produced kinins from HK. It is suggested that kinins, released at the Candida cell wall, can promote host colonization by the pathogen and the development of infection. PMID- 19804361 TI - Factors regulating tachyphylaxis triggered by N-terminal-modified angiotensin II analogs. AB - Binding of angiotensin II (DRVYIHPF, AngII) to its AT(1) receptor can trigger a process known as tachyphylaxis (loss of receptor response owing to repeated agonist stimulation). We propose a two-state binding model for tachyphylaxis where the N-terminal Asp(1) and Arg(2) residues of the peptide are supposed to initially bind to the N-terminal segment (Arg(23)) and to the EC-3 loop (Asp(281)) of an AT(1) molecule, respectively (state 1). Sequentially, a disruption of the salt bond between the AngII Asp(1) beta-carboxyl function and the receptor Arg(23) can occur with release of the peptide N-terminal segment, favoring the binding of the Arg(2) residue to the EC-3 loop (Asp(178,281), state 2). In the present study, we expanded this investigation by assaying pharmacological properties of different AngII analogs in guinea-pig ileum bearing modifications at positions 1 and 2. Most of these peptides were weak agonists but many of them had the ability to induce tachyphylaxis. These findings support the two-state model for tachyphylaxis, but alternative mechanisms were revealed where state 1 was no longer needed, depending on the chemical structure of AngII residue 1. Otherwise, any modification of the wild type AngII Arg(2) residue was deleterious for the tachyphylaxis mechanism. PMID- 19804362 TI - Hexose-6-phosphate dehydrogenase in the endoplasmic reticulum. AB - Hexose-6-phosphate dehydrogenase (H6PD) is a luminal enzyme of the endoplasmic reticulum that is distinguished from cytosolic glucose-6-phosphate dehydrogenase by several features. H6PD converts glucose-6-phosphate and NADP(+) to 6 phosphogluconate and NADPH, thereby catalyzing the first two reactions of the pentose-phosphate pathway. Because the endoplasmic reticulum has a separate pyridine nucleotide pool, H6PD provides NADPH for luminal reductases. One of these enzymes, 11beta-hydroxysteroid dehydrogenase type 1 responsible for prereceptorial activation of glucocorticoids, has been the focus of much attention as a probable factor in the pathomechanism of several human diseases including insulin resistance and the metabolic syndrome. This review summarizes recent advances related to the functions of H6PD. PMID- 19804363 TI - Chemokines in tumor-associated angiogenesis. AB - Tumor growth is dependent on several key factors. Apart from immune escape and an efficient blockade of apoptotic signals, tumors require oxygen and nutrients to grow past a diameter of 2 microm. Therefore, it is of vital importance for the tumor to facilitate tumor-associated angiogenesis, e.g., the de novo formation of new blood vessels. In addition to established and key angiogenic factors, such as vascular endothelial growth factor, chemokines, a superfamily of cytokine-like proteins that bind to seven transmembrane-spanning G-protein-coupled receptors, have been associated with angiogenesis under homeostatic conditions. Chemokines were initially identified as key factors that control the directional migration of leukocytes, stem cells and cancer cells in vitro and which critically regulate their trafficking in vivo. Recently their role in establishing a favorable microenvironment for tumor-associated angiogenesis, a process that requires complex bidirectional interactions of the tumor and associated vessels, has been the focus of research. Chemokine-promoted angiogenesis not only facilitates tumor growth by supplying nutrients and oxygen but it is also a prerequisite to tumor metastasis. Hence, the pharmacologic control of tumor angiogenesis presents a promising strategy for novel anticancer therapeutics. Here, we discuss the current pathogenetic concepts of tumor-associated angiogenesis in the context of chemokines and their receptors and highlight promising therapeutic strategies. PMID- 19804364 TI - Crystal structure of 4-hydroxybutyrate CoA-transferase from Clostridium aminobutyricum. AB - 4-Hydroxybutyrate CoA-transferases (4-HB-CoAT) takes part in the fermentation of 4-aminobutyrate to ammonia, acetate, and butyrate in anaerobic bacteria such as Clostridium aminobutyricum and Porphyromonas gingivalis or facultative anaerobic bacteria such as Shewanella oneidensis. Site-directed mutagenesis of the highly active enzyme has identified the catalytic glutamate residue as E238. Crystal structure of this enzyme has been determined at a resolution of 1.85 A. The 438 amino acid residue polypeptide chain folds into two topologically similar domains with an open alpha/beta-fold, which is also found in other CoAT family I and family II members. The data indicate that the members of CoAT families I and II are closely related; the latter only lacking the catalytic glutamate residue. A putative co-substrate binding site for the 4-HB-CoAT was identified, in which a 4 hydroxybutyrate molecule has been modeled. This site is also responsible for binding the acetyl group of acetyl-CoA or the succinyl group of succinyl-CoA in succinyl-CoA:3-oxoacid CoA-transferase from mammalian mitochondria. Mutations of relevant active site amino acid residues have been produced and their activities tested to corroborate the proposed structural model for substrate binding. 4-HB CoAT from C. aminobutyricum represents the only functionally characterized 4-HB CoAT present in the structural database. PMID- 19804365 TI - Cancer Osaka thyroid (Cot) phosphorylates Polo-like kinase (PLK1) at Ser137 but not at Thr210. AB - Cancer Osaka thyroid (Cot) is a proto-oncogenic kinase which belongs to the MAP3K family. A peptide-based substrate screening assay revealed that Cot has the ability to phosphorylate Polo-like kinase 1 (Plk1) at Ser137. Kinase assays with intact Plk1 and peptides surrounding Ser137 and Thr210 indicated further that Cot phosphorylates Ser137 but not Thr210. Additional support came from 3D peptide structure prediction and Cot-Plk1 interaction modeling. In vivo experiments demonstrated that wild type Cot, but not a kinase-dead mutant, has the ability to phosphorylate Ser137. Knockdown of Cot in Hela showed a reduction in the level of phosphorylation of Ser137. These results imply for the first time that Cot might be an upstream kinase of Plk1 and suggest a new mechanism for the regulation of the cellular function of Plk1. PMID- 19804366 TI - Coagulation factor XIII variants with altered thrombin activation rates. AB - Coagulation factor XIII (FXIII) is activated by thrombin and catalyses crosslinking between fibrin monomers thereby providing mechanical strength to the fibrin network. V34L is a common FXIII-A polymorphism found in the activation peptide. FXIII-A V34L is activated faster by thrombin and provides formation of a tighter clot at fibrinogen concentrations in the low end of the physiological range. FXIII-A variants with potentially increased activation rates were generated. Introduction of an optimal thrombin cleavage site, V34L+V35T, increased the activation rate 7.6-fold and facilitated the formation of a fibrin network more resistant to fibrinolysis than obtained with wt FXIII-A. In contrast, introduction of fragments of fibrinopeptide A into the activation peptide resulted in severely impaired activation rates. PMID- 19804368 TI - Cancer stem cells and the biology of brain tumors. AB - There is now compelling evidence that brain tumors harbor a small population of cells characterized by their ability to undergo self-renewal and initiate tumors, termed cancer stem cells (CSCs). The development of therapeutic strategies targeted towards CSC signaling may improve the treatment of brain tumors such as malignant gliomas and medulloblastomas. Here we review the role of cancer stem cells in glioma and medulloblastoma and some of the signaling mechanisms involved in brain tumor stem cell (BTSC) biology, and discuss how these signaling pathways may represent new stem cell targets for the treatment of brain tumors. In addition, we provide illustrative immunohistochemical data on the presence of BTSCs in human gliomas and medulloblastomas, and show preliminary findings suggesting the involvement of a GPCR, the gastrin-releasing peptide receptor (GRPR), in the expansion of BTSCs in vitro. PMID- 19804370 TI - DNA damage and repair in a model of rat vascular injury. AB - Restenosis rate following vascular interventions still limits their long-term success. Oxidative stress plays a relevant role in this pathophysiological phenomenon, but less attention has been devoted to its effects on DNA damage and to the subsequent mechanisms of repair. We analysed in a model of arteriotomy induced stenosis in rat carotids the time-dependent expression of DNA damage markers and of DNA repair genes, together with the assessment of proliferation and apoptosis indexes. The expression of the oxidative DNA damage marker 7,8 dihydro-8-oxo-2'-deoxyguanosine was increased at 3 and 7 days after arteriotomy, with immunostaining distributed in the injured vascular wall and in perivascular tissue. The expression of the DNA damage marker phospho-H2A.X was less relevant but increasing from 4 hrs to 7 days after arteriotomy, with immunostaining prevalently present in the adventitia and, to a lesser extent, in medial smooth muscle cells at the injury site. RT-PCR indicated a decrease of 8 out of 12 genes of the DNA repair machinery we selected from 4 hrs to 7 days after arteriotomy with the exception of increased Muyth and Slk genes (p<0.05). Western Blot revealed a decrease of p53 and catalase at 3 days after arteriotomy (p<0.05). A maximal 7% of BrdU-positive cells in endothelium and media occurred at 7 days after arteriotomy, while the apoptotic index peaked at 3 days after injury (p<0.05). Our results highlight a persistent DNA damage presumably related to a temporary decreased expression of the DNA repair machinery and of the antioxidant enzyme catalase, playing a role in stenosis progression. PMID- 19804371 TI - Coping with clean intermittent catherization--experiences from a patient perspective. AB - BACKGROUND: Poor bladder emptying is a well-known phenomenon in urology which increases the risk of urinary tract infection. Thus, it is important to insure effective bladder emptying for the patient. This may be performed by emptying the bladder intermittently by means of a disposable catheter. The method is called clean intermittent catheterization (CIC). The purpose of CIC is, apart from the scientific reasons, to give patients the opportunity to live the life they did prior to experiencing problems with bladder emptying. In our view, a large number of patients do not cope with their new life with CIC in spite of learning programmes and written guidance. AIM: The aim of this study was to develop knowledge about how patients experience having to empty their bladder by using a disposable catheter for the rest of their lives. Knowledge nurses could be used to improve the quality of the procedure for CIC training. Furthermore this study also helps in investigating the strategies used by the patients used to master CIC in order to improve the organization of training in the future. METHOD: The research design was qualitative and took a phenomenological hermeneutic approach. The data were based on interviews with eight patients using CIC. RESULTS AND CONCLUSION: The results suggested that need to perform CIC for the rest of one's life resulted in a traumatic crisis which could be experienced to be more or less stressful. Situations were mastered employing strategies which could either be problem- or emotion-focused. These strategies aimed, respectively, at solving the problem or at reducing the unpleasant feelings connected with the situation. PMID- 19804372 TI - User participation in community mental health services: exploring the experiences of users and professionals. AB - Increased user participation and community integration are central aims for contemporary mental health policy in many countries. User participation in community mental health services is developed through practice; from interaction between service-users and professionals working on the ground level. Despite this, there is a lack of research exploring users' and professionals' experiences and views based on the practice of user participation. The objective of this study was to illuminate user participation in a community mental health context based on the experiences of users and professionals within the same services. A qualitative study with an explorative design was applied. Preliminary data analyses based on a field study within three community mental health centres in a Norwegian city lead to our specific focus on experiences of user participation. This theme was explored in individual interviews with 10 users and two group interviews with six professionals. This article is based on the data from these interviews. All informants valued user participation in the service and highlighted the importance of the environment. Users and professionals did, however, highlight interesting issues of user participation from different perspectives. We developed the findings into three main themes: (i) user participation--experiences and preferences, (ii) an environment that promotes user participation and (iii) professional help, responsibility and user participation. Developing service-users' influence through participation is important, not only on the political and organisational level, but also in the contexts where users and professionals meet and collaborate. Self-determination in how to use services means that there are opportunities for receiving support without being subjected to control. Community mental health services which provide flexible, accepting environments with possibilities for both support and challenges may enhance participation and give all users possibilities to have an influence. PMID- 19804369 TI - Cartilage tissue engineering: towards a biomaterial-assisted mesenchymal stem cell therapy. AB - Injuries to articular cartilage are one of the most challenging issues of musculoskeletal medicine due to the poor intrinsic ability of this tissue for repair. Despite progress in orthopaedic surgery, the lack of efficient modalities of treatment for large chondral defects has prompted research on tissue engineering combining chondrogenic cells, scaffold materials and environmental factors. The aim of this review is to focus on the recent advances made in exploiting the potentials of cell therapy for cartilage engineering. These include: 1) defining the best cell candidates between chondrocytes or multipotent progenitor cells, such as multipotent mesenchymal stromal cells (MSC), in terms of readily available sources for isolation, expansion and repair potential; 2) engineering biocompatible and biodegradable natural or artificial matrix scaffolds as cell carriers, chondrogenic factors releasing factories and supports for defect filling, 3) identifying more specific growth factors and the appropriate scheme of application that will promote both chondrogenic differentiation and then maintain the differentiated phenotype overtime and 4) evaluating the optimal combinations that will answer to the functional demand placed upon cartilage tissue replacement in animal models and in clinics. Finally, some of the major obstacles generally encountered in cartilage engineering are discussed as well as future trends to overcome these limiting issues for clinical applications. PMID- 19804373 TI - Coping with myocardial infarction: evaluation of a coping questionnaire. AB - The negative effects of emotional distress on the recovery following myocardial infarction make it important to study coping strategies in this situation. The present study aimed to evaluate the psychometric properties and the validity of a 10 dimensions questionnaire labelled The General Coping Questionnaire (GCQ). The structure of the questionnaire was based on a previous interview study with 26 persons with different diseases. The 10 dimensions are called self-trust, problem reducing actions, change of values, social trust, minimization, fatalism, resignation, protest, isolation and intrusion. The present study comprised 114 first-time myocardial infarction patients (37 women, 77 men). Five months after myocardial infarction, they answered questions about health-related quality of life, health complaints, sense of coherence and the GCQ. A multi-trait/multi-item analysis showed good item-scale convergent and discriminatory validity when the GCQ was reduced from 47 to 40 items. In conclusion, the results showed that the 40-item GCQ is a well-structured and reliable questionnaire for measuring coping strategies in myocardial infarction patients. PMID- 19804374 TI - Being in charge of life: perceptions of lifestyle among women of retirement age. AB - AIM: The aim of this study was to describe the perceptions of lifestyle among women of retirement age. BACKGROUND: Women go through many transitions during their lives, which impact on their lifestyle and possibly their outlook on life. Family circumstances such as motherhood and marital status change over time and the menopause is also likely to influence their view of life. METHOD: Data were collected through interviews with 20 women, aged 61-70, selected by means of strategic sampling. The interviews were analysed using a phenomenographic approach. FINDINGS: Three structural aspects emerged: being healthy as life turns out, living life in their own way, and taking care of everyday life. The women described lifestyle as a means of being healthy, having an active role in society, being content with what one has and the need to adapt oneself to limiting circumstances. Lifestyle was also associated with being in charge and making one's own choices in life based on one's own values. They reported that they were independent and made decisions about their life. The informants considered that lifestyle was an asset that helped them to cope with everyday life and to make the most of each day. CONCLUSION: Lifestyle is a tool that requires independence, the right to self-determination over everyday life and adjustment to limitations in order to ensure the psychological well-being of women of retirement age. Further research is needed in order to delineate the possible association between lifestyle and psychological well-being. PMID- 19804375 TI - Self-rated health--what does it capture at 1 year after childbirth? Investigation of a survey question employing thinkaloud interviews. AB - AIM: This paper reports an investigation of how the survey question 'How would you summarize your state of health at present' is interpreted and what it captures when asked at 1 year after childbirth. BACKGROUND: Self-rated health measured by a single item question is a well-established patient outcome as it predicts morbidity and the use of health services. However, there is limited understanding of what the question captures in early motherhood. METHOD: A qualitative design combining data collection by means of a short form, concurrent and retrospective thinkaloud interviews, and a semi-structured interview, with 26 Swedish women during 2005 was employed. The text was analysed by qualitative content analysis. A theoretical framework describing four cognitive tasks usually performed when a respondent answers a survey question guided the analysis: interpretation of the question, retrieval of information, forming a judgement and giving a response. FINDINGS: The questions of self-rated health left open for the new mothers to evaluate what was most important for her. It captured a woman's total life situation, such as family functioning and well-being, relationship with partner, combining motherhood and professional work, energy, physical symptoms and emotional problems affecting daily life, stressful life events, chronic disease with ongoing symptoms, body image, physical exercise and happiness. Neither childbirth-related events nor childbirth-related symptoms were included in the responses. Less than 'good' self-rated health represented a high burden of health problems. CONCLUSION: Our results showed that the question on self-rated health was a measure of women's general health and well-being in their present life situation, but it did not seem to measure recovery after childbirth specifically. PMID- 19804376 TI - Patterns of colonization and spread in the fungal spruce pathogen Onnia tomentosa. AB - The basidiomycetous fungus Onnia tomentosa is one of the most widespread root rot pathogens in North America. Although the disease is more severe on spruce and pine trees, this pathogen can infect several coniferous species. To study the population structure of O. tomentosa, we harvested 180 basidiocarps in a 45-year old white spruce plantation in western Quebec in autumn 1997 and extracted DNA directly from individual basidiocarps. Using a combination of spatial coordinates and molecular data based on the analysis of two mitochondrial and three nuclear loci, we measured the average genet size and molecular diversity and assessed the relative contribution of basidiospores and vegetative growth to the stand colonization. Most of the sampled basidiocarps that clustered spatially belonged to the same genet. A total of 37 discrete multilocus genets of an average size of 3.42 m were obtained. The genet size distribution was skewed towards smaller genets (<3 m) that displayed higher diversity than the larger genets (>3 m). The nuclear loci were in Hardy-Weinberg equilibrium in the larger genets, but not in the smaller genets, which displayed a deficiency of heterozygotes. This suggests a Wahlund effect, whereby different colonization events resulted in expected heterozygosity higher than observed heterozygosity. Using an estimate of the growth rate of the fungus, only a few of the largest genets were approximately the age of the plantation. These observations are consistent with the colonization by basidiospores subsequent to site preparation and tree planting followed by secondary colonization events and vegetative spread. PMID- 19804377 TI - Multiyear multiple paternity and mate fidelity in the American alligator, Alligator mississippiensis. AB - We examined multiple paternity during eight breeding events within a 10-year period (1995-2005) for a total of 114 wild American alligator nests in Rockefeller Wildlife Refuge in south-west Louisiana. Our goals included examining (i) within population variation in multiple paternity among years, (ii) variation in multiple paternity in individual females and (iii) the potential for mate fidelity. To accomplish this, in the current study, eggs were sampled from 92 nests over 6 years and analysed along with 22 nests from a previous 2-year study. Genotypes at five microsatellite loci were generated for 1802 alligator hatchlings. Multiple paternity was found in 51% of clutches and paternal contributions to these clutches were highly skewed. Rates of multiple paternity varied widely among years and were consistently higher in the current study than previously reported for the same population. Larger females have larger clutches, but are not more likely to have multiply sired nests. However, small females are unlikely to have clutches with more than two sires. For 10 females, nests from multiple years were examined. Seven (70%) of these females exhibited long-term mate fidelity, with one female mating with the same male in 1997, 2002 and 2005. Five females exhibiting partial mate fidelity (71%) had at least one multiple paternity nest and thus mated with the same male, but not exclusively. These patterns of mate fidelity suggest a potential role for mate choice in alligators. PMID- 19804378 TI - Nuclear and mitochondrial DNA reveals isolation of imperilled grey nurse shark populations (Carcharias taurus). AB - Loss of sharks and other upper-trophic marine predators has sparked worldwide concern for the stability of ocean ecosystems. The grey nurse (ragged-tooth or sand tiger) shark (Carcharias taurus) is Vulnerable on a global scale, Critically Endangered in Australia and presumed extinct in parts of its historical range. We used 193 muscle and fin samples collected from six extant populations to assess global mtDNA and microsatellite diversity and the degree of global population genetic structure. Control region mtDNA diversity was low in every population, and two populations (eastern Australia and Japan) contained only a single mtDNA haplotype. Genetic signatures of recent losses of genetic variation were not yet apparent at microsatellite loci, indicating that this low mtDNA variation is not a result of anthropogenic population declines. Population differentiation was substantial between each population pair except Brazil and South Africa, F(ST) values ranged from 0.050 to 0.699 and 0.100 to 1.00 for microsatellite and mitochondrial data respectively. Bayesian analysis clearly partitioned individuals into five of the populations from which they were sampled. Our data imply a low frequency of immigrant exchange among each of these regions and we suggest that each be recognized as a distinct evolutionary significant unit. In contrast to pelagic species such as whale shark and white shark that may cross ocean basins and where cooperative international efforts are necessary for conservation, grey nurse shark, like many coastal species, need to be managed regionally. PMID- 19804379 TI - alpha-secretase mediated conversion of the amyloid precursor protein derived membrane stub C99 to C83 limits Abeta generation. AB - The Swedish mutation within the amyloid precursor protein (APP) causes early onset Alzheimer's disease due to increased cleavage of APP by BACE1. While beta secretase shedding of Swedish APP (APPswe) largely results from an activity localized in the late secretory pathway, cleavage of wild-type APP occurs mainly in endocytic compartments. However, we show that liberation of Abeta from APPswe is still dependent on functional internalization from the cell surface. Inspite the unchanged overall beta-secretase cleaved soluble APP released from APP(swe) secretion, mutations of the APPswe internalization motif strongly reduced C99 levels and substantially decreased Abeta secretion. We point out that alpha secretase activity-mediated conversion of C99 to C83 is the main cause of this Abeta reduction. Furthermore, we demonstrate that alpha-secretase cleavage of C99 even contributes to the reduction of Abeta secretion of internalization deficient wild-type APP. Therefore, inhibition of alpha-secretase cleavage increased Abeta secretion through diminished conversion of C99 to C83 in APP695, APP695swe or C99 expressing cells. PMID- 19804380 TI - Relative positioning of diazepam in the benzodiazepine-binding-pocket of GABA receptors. AB - GABA(A) receptors are the major inhibitory neurotransmitter receptors in the brain. Some of them are targets of benzodiazepines that are widely used in clinical practice for their sedative/hypnotic, anxiolytic, muscle relaxant and anticonvulsant effects. In order to rationally separate these different drug actions, we need to understand the interaction of such compounds with the benzodiazepine-binding pocket. With this aim, we mutated residues located in the benzodiazepine-binding site individually to cysteine. These mutated receptors were combined with benzodiazepine site ligands carrying a cysteine reactive group in a defined position. Proximal apposition of reaction partners will lead to a covalent reaction. We describe here such proximity-accelerated chemical coupling reactions of alpha(1)S205C and alpha(1)T206C with a diazepam derivative modified at the C-3 position with a reactive isothiocyanate group (-NCS). We also provide new data that identify alpha(1)H101C and alpha(1)N102C as exclusive sites of the reaction of a diazepam derivative where the -Cl atom is replaced by a -NCS group. Based on these observations we propose a relative positioning of diazepam within the benzodiazepine-binding site of alpha(1)beta(2)gamma(2) receptors. PMID- 19804381 TI - Diurnal cortisol secretion at home and in child care: a prospective study of 2 year-old toddlers. AB - BACKGROUND: Previous studies indicate that children may experience disrupted cortisol secretion in child care. The extent to which this is a transient or long term disruption is not known, as most studies have relied on cross-sectional designs, and age-heterogeneous small sample sizes. This study aims to (a) compare cortisol secretion measured at home and in child care at 2 and 3 years of age, (b) investigate cortisol changes from 2 to 3 years of age, (c) examine whether age at initiation of child care is associated with cortisol secretion, and (d) investigate whether cortisol secretion in child care is linked to behavioural problems. METHODS: Saliva samples were collected in a cohort of children recruited at 2 years of age from a larger population sample composed of women seen for the first time during pregnancy. Saliva was sampled twice a day (morning and afternoon) over two consecutive days at home and in child care at 2 (n = 155) and 3 years of age (n = 116). Interviews regarding the familial socioeconomic background and child care history were conducted with the mothers. RESULTS: At 2 years of age, children showed a flat diurnal cortisol pattern in child care and a decreasing pattern at home. At age 3 years, children showed decreasing patterns both at home and in child care. Also at 3 years, children with less child care experience (i.e., entry after 16 months) had higher cortisol levels in child care and lower levels at home. In contrast, those with more experience (i.e., entry prior to 8 months) had lower cortisol in child care and higher cortisol at home. CONCLUSION: The different patterns of diurnal secretion observed in child care as compared to home is transient for most children, diminishing as they get older, whereas home and child care overall levels later on may be influenced by the cumulated experience with child care. PMID- 19804382 TI - Area and family effects on the psychopathology of the Millennium Cohort Study children and their older siblings. AB - BACKGROUND: To model and compare contextual (area and family) effects on the psychopathology of children nested in families nested in areas. METHOD: Data from the first two sweeps of the UK's Millennium Cohort Study were used. The final study sample was 9,630 children clustered in 6,052 families clustered in 1,681 Lower-layer Super Output Areas. The mean age of the children at Sweep 2 was 4.96 (SD = 2.76) years. Contextual risk was measured at area level with the Index of Multiple Deprivation (Sweep 1), and at family level with the number of proximal (Sweep 2) and distal (Sweep 1) adverse life events experienced. Psychopathology was measured at Sweep 2 with the Strengths and Difficulties Questionnaire. RESULTS: At baseline, both proximal and distal family risk and area risk predicted broad psychopathology, although the most parsimonious was the proximal family risk model, and both the family-level and the area-level variability were significant. The area risk/broad psychopathology association remained significant even when family risk was controlled, but not when family socioeconomic status was controlled. The full model added parenting and paternal and maternal psychopathology. When parental qualifications were excluded from the family-level contextual controls the effect of area risk remained significant on both externalizing and internalizing psychopathology. CONCLUSIONS: The effect of area on child psychopathology operated via the socioeconomic characteristics of the child's family, not just the adverse characteristics of the neighbors. Multiple family risk predicted child psychopathology directly and independently, and not because it was associated with family socioeconomic status. Family socioeconomic status explained the association between area risk and broad psychopathology. PMID- 19804383 TI - Effectiveness of a balance training home exercise programme for adults with haemophilia: a pilot study. AB - Adults with haemophilia and other bleeding disorders often develop lower limb musculoskeletal problems associated with bleeds into joints and muscles, which may affect balance performance and increase likelihood of falling. The aim of this study was to evaluate the effectiveness of an individualized balance and strength home exercise programme on improving balance and related outcomes for adults with haemophilia and other bleeding disorders. Twenty male adults with haemophilia and other bleeding disorders (mean age 39.4 years, 95% CI = 33.7 45.1) were recruited to participate. They underwent a comprehensive clinical and force platform assessment of balance and related measures. Based on assessment findings, the assessing physiotherapist provided an individualized home exercise programme of balance, strengthening and walking exercises. Re-assessment occurred after the 4-month exercise programme. Twelve participants (60%) completed the programme and were re-assessed. There were no safety problems or dropouts associated with the exercise programme aggravating joint status. Although there were no statistically significant changes in any of the measures (adjusted for multiple comparisons), there were improvements of between 5% and 22% on 10 of the 16 measures, with the Neurocom modified Clinical Test of Sensory Interaction on Balance (P = 0.036) and Timed Sit to Stand (P = 0.064) approaching significance. A tailored home exercise programme targeting balance, strengthening and walking is feasible for adults with haemophilia and other bleeding disorders. These results suggest that positive physical outcomes including improved balance and mobility may be achieved with this type of programme. PMID- 19804384 TI - Influence of aquatic training on the motor performance of patients with haemophilic arthropathy. AB - Thirteen patients with haemophilia A took part in this study voluntarily. They underwent an aquatic training programme over a 9-week period (27 sessions; three sessions per week; 1 h per session). Their motor performance was assessed by the following cardio-respiratory and mechanical variables before and after the training programme: oxygen uptake (VO(2), mL min(-1)), relative oxygen uptake (rel VO(2), mL min(-1).kg(-1)), carbon dioxide (CO(2), mL min(-1)), respiratory quotient (R), heart rate (bpm) and the distance covered in 12 min (the Cooper test, m). Nine patients successfully completed the intervention and measurement protocols without bleeding or other adverse events. After the proposed training programme, significant differences between the pre-test and post-test were observed. Patients' aerobic capacity increased considerably, and their oxygen uptake improved by 51.51% (P < 0.05), while their relative oxygen uptake went up by 37.73% (P < 0.05). Their mechanical capacity also increased considerably (14.68%, P < 0.01). Our results suggest that 27 specially designed aquatic training sessions for our patients with haemophilia A had a positive effect on their motor performance and considerably improved their aerobic and mechanical capacity without causing adverse effects. PMID- 19804385 TI - Sorafenib as a feasible therapeutic option in haemophiliacs with hepatocellular carcinoma. PMID- 19804386 TI - Prosthetic heart valves and rare hypersensitivity to vitamin K antagonists resulting from factor IX mutation: how to manage anticoagulation? PMID- 19804387 TI - Effectiveness of nonpharmacologic treatment for migraine in young children. AB - OBJECTIVE: To evaluate the effectiveness of nonpharmacologic treatment for migraine in children younger than age 6 years. BACKGROUND: The mean age of onset of migraine in children is 7.2 years for boys and 10.9 years for girls. Treatment consists of individually tailored pharmacologic and nonpharmacologic interventions. However, data on migraine management in preschoolers are very sparse. METHODS: Demographic, clinical, and outcome data were collected from the files of patients with migraine who attended a pediatric headache clinic. Only those treated by nonpharmacologic measures, namely, good sleep hygiene, diet free of food additives, and limited sun exposure, were included. Clinical factors and response to treatment were compared between children younger than 6 years and older children. RESULTS: Of the 92 children identified, 32 were younger than 6 years and 60 were older. There was no difference between the age groups in most of the demographic and clinical parameters. The younger group was characterized by a significantly lower frequency of migraine attacks and shorter disease duration (in months). Mean age of the patients with no response to treatment (grade 1) was 10.588 +/- 3.254 years; partial response (grade 2), 9.11 +/- 4.6 years; and complete response (grade 3), 8.11 +/- 3.93 years (P = .02). The percentage of patients with complete to partial response as opposed to no response was significantly higher in the younger group (P = .00075). CONCLUSION: As the primary option, conservative therapy for migraine appears to be more effective in children younger than 6 years than in older children, perhaps because of their shorter duration of disease until treatment and lower frequency of attacks. PMID- 19804388 TI - Increased dopamine is associated with the cGMP and homocysteine pathway in female migraineurs. AB - BACKGROUND: The group of catecholamines, which include dopamine, adrenaline, and noradrenaline, are neurotransmitters which have been considered to play a role in the pathogenesis of migraine. However, the impact of catecholamines, especially dopamine on migraine as well as the exact mechanisms is not clear to date as previous studies have yielded in part conflicting results. OBJECTIVE: This study aimed to produce a comprehensive examination of dopamine in migraineurs. METHODS: Catecholamines and various parameters of the homocysteine, folate, and iron metabolism as well as cyclic guanosine monophosphate (cGMP) and inflammatory markers were determined in 135 subjects. RESULTS: We found increased dopamine levels in the headache free period in female migraineurs but not in male patients. Increased dopamine is associated with a 3.30-fold higher risk for migraine in women. We found no significant effects of aura symptoms or menstrual cycle phases on dopamine levels. Dopamine is strongly correlated with cGMP and the homocysteine-folate pathway. CONCLUSION: We show here that female migraineurs exhibit increased dopamine levels in the headache free period which are associated with a higher risk for migraine. PMID- 19804389 TI - Headache disability among adolescents: a student population-based study. AB - OBJECTIVE: To examine the extent and to identify the relevant predictors of headache disabilities in adolescents. BACKGROUND: Headaches are common in adolescents but their impact and related factors have not been extensively studied in adolescent communities. METHOD: We recruited and surveyed 3963 students aged 13-15 from 3 middle schools using self-administered questionnaires. The questionnaires were used to make 3 assessments: (1) headaches were diagnosed using a validated headache questionnaire; (2) headache disabilities were evaluated using the 6-question Pediatric Migraine Disability Assessment; (3) depression was measured using the Adolescent Depression Inventory. RESULTS: The student response rate was 93%. In total, 484 students (12.2%) had migraines with or without auras, 444 (11.2%) had probable migraines, and 1092 (27.6%) had tension-type headaches. The students with migraine had the highest Pediatric Migraine Disability Assessment scores (10.7 +/- 20.0); whereas, the students with tension-type headaches had the lowest scores (2.0 +/- 4.4). Logistic regression analyses indicated that there were a number of independent predictors for moderate to severe headache-related disability (Pediatric Migraine Disability Assessment score > or =31), including a migraine or probable migraine diagnosis, a higher depression score, severe headache intensity, and frequent headaches. CONCLUSIONS: The Pediatric Migraine Disability Assessment provides a simple tool to measure the impact of headaches in adolescents. Adolescents with migraine headaches suffered the greatest level of disability. Higher depression scores were associated with more severe headache-related disabilities in adolescents, independent of headache frequency and severity. PMID- 19804390 TI - The safety of sumatriptan and naratriptan in pregnancy: what have we learned? AB - OBJECTIVES: To monitor for a signal of major teratogenicity by determining the risk of all major defects following in utero exposure to sumatriptan and naratriptan. To monitor for unusual patterns of birth defects that might suggest teratogenicity. BACKGROUND: The prevalence of migraine is highest in women of childbearing age. Coupled with the recurrent nature of migraine attacks and the high proportion of unplanned pregnancies, intentional and inadvertent exposure to these drugs in pregnancy is likely. The Sumatriptan and Naratriptan Pregnancy Registry captures data on women exposed to those drugs during pregnancy to monitor for evidence of major teratogenicity. METHODS: Healthcare professionals from anywhere in the world can enroll, on a voluntary basis, women exposed to sumatriptan or naratriptan during their pregnancies in this primarily prospective, observational study. Only pregnancies with unknown outcomes at the time of enrollment were included in the analysis. The percentage of infants or fetuses with major birth defects was calculated as the total number of infants/fetuses with major birth defects divided by the sum of the number of infants/fetuses with major birth defects + the number of live births without defects. The risk of major birth defects was further stratified by earliest trimester of pregnancy exposure. RESULTS: Data are available on pregnancy outcomes from 599 exposed women. Among 479 first-trimester exposures to sumatriptan, 20 outcomes with major birth defects were reported (4.6%, 95% confidence interval [CI] 2.9-7.2%). The risk of major birth defects following exposure to sumatriptan during any trimester was 4.7% (95% CI 3.1-7.0%). No distinctive pattern of major birth defects among exposed infants was noted. There were 50 first-trimester exposures to naratriptan with 1 reported birth defect in a fetus with exposure to both sumatriptan and naratriptan. CONCLUSIONS: The risk of all major birth defects following first-trimester exposure to sumatriptan was 4.6% (95% CI 2.9-7.2%). This coupled with a consistent failure of additional epidemiological studies to observe a signal for major teratogenicity gives a level of reassurance concerning the safety of sumatriptan in pregnancy. There are too few data on naratriptan to draw definitive conclusions, and the sample size for sumatriptan remains too small to detect any but very large increases in specific birth defects. PMID- 19804391 TI - Prevalence and incidence of headache in adolescent Finnish twins. AB - OBJECTIVES: The study aim was to determinate changes in prevalence and incidence rates of headache among adolescent Finnish twins. METHODS: Questionnaire data were collected during 1994-1998 from a nationwide sample of Finnish families of 11-year-old twins who were born between 1983 and 1987 (n = 6262) and again at ages of 14 and 17. RESULTS: During follow-up, the prevalence of regular headache (ie, at least once a month = RHA) increased from 59% at age 11 to 65% at age 14 and decreased to 63% at age 17. Among boys the prevalence of RHA increased significantly from 59% at age 11 to 62% at age 14 decreasing to 52% at age 17. Among girls the prevalence rates increased significantly between ages of 11 and 14 from 60% to 68% up to 74% at age 17. The prevalence of weekly headache increased in girls between ages of 11 and 14 from 16% to 25%. Incidence of RHA (at least once a month) decreased from 47% at age 14 to 44% at age 17 in girls and from 44% at age 14 to 30% at age 17 in boys. CONCLUSIONS: RHA is more common in girls than boys during puberty, while the incidence of RHA declined especially in boys during same age period. PMID- 19804393 TI - Endonasal endoscopic management of contact point headache and diagnostic criteria. AB - BACKGROUND: Some types of headaches with sinonasal origin may be present in the absence of inflammation and infection. The contact points between the lateral nasal wall and the septum could be the cause of triggering and sustained pain via trigeminovascular system. OBJECTIVE: The aim of this study was to evaluate the feasibility and effectiveness of endoscopic surgery in the sinonasal region for treatment of headache with special attention paid to specific diagnostic methods and patient selection. METHODS: This was a prospective, non-randomized and semi quasi experimental research study. Thirty-six patients with chronic headaches who had not previously responded to conventional treatments were evaluated by rhinoscopy and/or endoscopy, local anesthetic tests and computed tomography scans as diagnostic criteria. These patients were divided into 4 groups based on the diagnostic methods utilized. The intensity of headaches pre- and post-operatively were recorded by utilizing the visual analog scale scale and performing analysis with analysis of variance test comparison and Statistical Package for Social Sciences. Average follow-up was 30 months. RESULTS: Our overall success rate approximated 83% while the complete cure rate was 11%. Patients in group 4 achieved the best results. In this group all diagnostic criteria were positive. In addition, patient responses were statistically significant in groups with more than one positive criteria compared with group 1 who only had positive examination. The positive response of 14 migrainous patients diagnosed with migraine prior to treatment was 64%. CONCLUSION: Surgery in specific cases of headaches with more positive evidence of contact point could be successful, particularly if medical therapy has failed. PMID- 19804392 TI - Topiramate for migraine prevention in a naturalistic setting: results from an open label, flexible dose study. AB - BACKGROUND: Headaches are one of the most common neurological symptoms and migraines are the most common primary headache disorder. The global prevalence of migraines is around 10% and the condition is associated with a high burden of disease. Despite an abundance of good quality evidence, only 1 in 5 of patients who fulfill the criteria for preventive migraine therapy are appropriately treated. Data on patient outcomes with preventive medication derive mostly from specialized academic centers, which contrasts with normal clinical practice where the majority of patients are treated outside tertiary care centers. OBJECTIVE: To explore tolerability, safety and efficacy outcomes of patients receiving topiramate for migraine prevention in a naturalistic setting. METHODS: After a 4 week prospective baseline, patients with a diagnosis of migraine according to International Headache Society criteria and eligible for migraine prevention were treated with flexible dosing of topiramate for 24 weeks (core phase), and optionally for a total of 48 weeks. The primary safety analysis included adverse events (AEs) during the core phase. For the main efficacy measures, the absolute changes from baseline to end of core phase as well as last follow-up visit were calculated for migraine days per 4 weeks, migraine attacks per 4 weeks, mean maximum visual analogue scale of migraine headache per 4 weeks and mean maximum pain intensity of migraine headache (4-point scale) per 4 weeks. In addition, changes in individual quality of life aspects were captured. RESULTS: The intention-to-treat population (ITT) consisted of 161 patients (90.7% female, mean age 45.7 +/- 11.1 years). Topiramate median dose was 45.7 mg/day at endpoint. Some 74.1% of patients reported treatment emergent AEs, most frequently paresthesias (18.4%) and nausea (12.4%). Some 20.0% of patients withdrew from the study due to AEs. The mean number of migraine days per 4 week decreased from 6.2 +/- 3.9 days at baseline to 3.9 +/- 3.5 days at last core visit (P < .001). Mean maximum pain intensity per 4 week changed from 7.0 +/- 2.3 at baseline to 4.7 +/- 3.2 at last visit core phase (P < .001). Consumption of triptans and analgesics reduced during the course of the core phase (P < .005). Fifty-one percent of all patients experienced at least a 50% reduction in migraine days during the core phase. CONCLUSION: Topiramate used for migraine prevention in non-academic institutions is generally safe, well tolerated and results in good control of migraine headaches and improvement in several aspects of quality of life. PMID- 19804394 TI - Cluster headache as the result of secondhand cigarette smoke exposure during childhood. AB - Unique to cluster headache (CH) compared with all other primary headache conditions is its association with a personal history of cigarette smoking. Studies have indicated that greater than 80% of CH patients have a prolonged history of tobacco usage prior to CH onset. How tobacco exposure can lead to CH has not yet been elucidated. As secondhand smoke exposure during childhood has been linked to multiple medical illnesses could CH also be the result of childhood exposure to tobacco smoke? The United States Cluster Headache survey is the largest survey ever done of CH sufferers living in the United States. The survey addressed various clinical, epidemiologic, and economic issues related to CH. Several survey questions dealt with the issue of personal and parental smoking history. Results from the survey suggest that CH can result from secondhand cigarette smoke exposure during childhood as greater than 60% of non smoking CH patients had parents who smoked. Strengthening the probable association between secondhand smoke exposure and the development of CH is the fact that double the number of survey responders developed CH at or before 20 years of age if during their childhood they lived with a parent who smoked cigarettes. PMID- 19804395 TI - Migraine is common comorbidity in Japanese patients with multiple sclerosis. PMID- 19804396 TI - Incomplete Posterior Circle of Willis in Migraineurs With Aura. PMID- 19804397 TI - Migraine and obesity: cause or effect? PMID- 19804398 TI - Association of hemicrania continua and temporomandibular dysfunction: the role of each team player. PMID- 19804399 TI - Numb chin syndrome, migraine-like headache and leptomeningeal spread of a B-cell lymphoma. PMID- 19804400 TI - Ankle-brachial index, a screening for peripheral obstructive arterial disease, and migraine - a controlled study. AB - BACKGROUND: Epidemiological studies support the association between migraine, especially migraine with aura, and vascular disorders. The ankle-brachial index (ABI) is largely used as a surrogate of peripheral obstructive arterial disorders (POAD). Accordingly, in this study we contrasted the ABI in individuals with migraine and in controls. METHODS: We investigated 50 migraineurs and 38 controls and obtained the ABI (ratio between the systolic arterial pressure obtained in the legs and in the arms) using digital sphygmomanometry. As per validation studies, we used the cut-off of 0.9 as the normal limit for the ABI. We adjusted for gender, use of contraceptive hormones, tabagism, and other cardiovascular risk factors. RESULTS: We found abnormal values of ABI, suggestive of mild or moderate POAD, in 31 individuals (35.2%). Mean value was 0.96 (standard deviation = 0.10). None of our patients had ABI < 0.4, which would suggest severe POAD. Mean ABI for migraineurs was 0.94 (0.11), and for controls it was 0.99 (0.09). Difference was significant (t = 2.21 and P = .022). After adjustments, ABI remained significantly associated with migraine status (P = .024). Adjustments were reasonably effective (chi(2) of Hosmer-Lemeshow = 1.06, P = .590). CONCLUSION: Our findings suggest that decreased values of ABI are more common in migraineurs than in controls. Although causality was not assessed by us, the relationship is of importance per se. Doctors should measure the ABI in individuals with migraine as an easy way to screen for cardiovascular risk. PMID- 19804401 TI - Fluctuating selection and the maintenance of individual and sex-specific diet specialization in free-living oystercatchers. AB - Fluctuating and disruptive selection are important mechanisms for maintaining intrapopulation trait variation. Nonetheless, few field studies quantify selection pressures over long periods and identify what causes them to fluctuate. Diet specialists in oystercatchers differ in short-term payoffs (intake), but their long-term payoffs are hypothesized to be condition dependent. We test whether phenotypic selection on diet specialization fluctuates between years due to the frequency of specialists, competitor density, prey abundance, and environmental conditions. Short-term payoffs proved to be poor predictors of long term fitness payoffs of specialization. Sex-differences in diet specialization were maintained by opposing directional fecundity and viability selection between the sexes. Contrasting other studies, selection on individual diet specialization was neither negative frequency- or density-dependent nor dependent on prey abundance. Notwithstanding, viability selection fluctuated strongly (stabilizing< ->disruptive) over the 26-year study period: slightly favoring generalists in most years, but strongly disfavoring generalists in rare harsh winters, suggesting generalists cannot cope with extreme conditions. Although selection fluctuated, mean selection on specialists was weak, which can explain how individual specialization can persist over long periods. Because rare events can dramatically affect long-term selective landscapes, more care should be taken to match the timescale of evolutionary studies to the temporal variability of critical environmental conditions. PMID- 19804402 TI - Morphological and molecular evidence reveals recent hybridization between gorilla taxa. AB - Molecular studies have demonstrated a deep lineage split between the two gorilla species, as well as divisions within these taxa; estimates place this divergence in the mid-Pleistocene, with gene flow continuing until approximately 80,000 years ago. Here, we present analyses of skeletal data indicating the presence of substantial recent gene flow among gorillas at all taxonomic levels: between populations, subspecies, and species. Complementary analyses of DNA sequence variation suggest that low-level migration occurred primarily in a westerly-to easterly direction. In western gorillas, the locations of hybrid phenotypes map closely to expectations based on population refugia and riverine barrier hypotheses, supporting the presence of significant vicariance-driven structuring and occasional admixture within this taxon. In eastern lowland gorillas, the high frequency of hybrid phenotypes is surprising, suggesting that this region represents a zone of introgression between eastern gorillas and migrants from the west, and underscoring the conservation priority of this critically endangered group. These results highlight the complex nature of evolutionary divergence in this genus, indicate that historical gene flow has played a major role in structuring gorilla diversity, and demonstrate that our understanding of the evolutionary processes responsible for shaping biodiversity can benefit immensely from consideration of morphological and molecular data in conjunction. PMID- 19804403 TI - Adaptive radiations, ecological specialization, and the evolutionary integration of complex morphological structures. AB - The evolutionary integration of complex morphological structures is a macroevolutionary pattern in which morphogenetic components evolve in a coordinated fashion, which can result from the interplay among processes of developmental, genetic integration, and different types of selection. We tested hypotheses of ecological versus developmental factors underlying patterns of within-species and evolutionary integration in the mandible of phyllostomid bats, during the most impressive ecological and morphological radiation among mammals. Shape variation of mandibular morphogenetic components was associated with diet, and the transition of integration patterns from developmental to within-species to evolutionary was examined. Within-species (as a proxy to genetic) integration in different lineages resembled developmental integration regardless of diet specialization, however, evolutionary integration patterns reflected selection in different mandibular components. For dietary specializations requiring extensive functional changes in mastication patterns or biting, such as frugivores and sanguivores, the evolutionary integration pattern was not associated with expected within-species or developmental integration. On the other hand, specializations with lower mastication demands or without major functional reorganization (such as nectarivores and carnivores), presented evolutionary integration patterns similar to the expected developmental pattern. These results show that evolutionary integration patterns are largely a result of independent selection on specific components regardless of developmental modules. PMID- 19804404 TI - Diversification of the African genus Protea (Proteaceae) in the Cape biodiversity hotspot and beyond: equal rates in different biomes. AB - The Cape region of South Africa is a hotspot of flowering plant biodiversity. However, the reasons why levels of diversity and endemism are so high remain obscure. Here, we reconstructed phylogenetic relationships among species in the genus Protea, which has its center of species richness and endemism in the Cape, but also extends through tropical Africa as far as Eritrea and Angola. Contrary to previous views, the Cape is identified as the ancestral area for the radiation of the extant lineages: most species in subtropical and tropical Africa are derived from a single invasion of that region. Moreover, diversification rates have been similar within and outside the Cape region. Migration out of the Cape has opened up vast areas, but those lineages have not diversified as extensively at fine spatial scales as lineages in the Cape. Therefore, higher net rates of diversification do not explain the high diversity and endemism of Protea in the Cape. Instead, understanding why the Cape is so diverse requires an explanation for how Cape species are able to diverge and persist at such small spatial scales. PMID- 19804405 TI - Associations of the IL-1 and TNF gene polymorphisms in the susceptibility to duodenal ulcer disease in Chinese Han population. AB - Our aim was to investigate whether genetic polymorphism of IL-1Beta-511, IL-1RN, TNF-A-308 are involved in the susceptibility to duodenal ulcer (DU). 437 unrelated Chinese Han patients with DU and 148 healthy controls were genotyped by the polymerase chain reaction-restriction fragment length polymorphism method for the IL-1B-511, TNF-A-308 gene polymorphisms and the VNTR polymorphism in intron 2 of the IL-1RN gene polymorphisms. There was no difference in the genetic polymorphism of IL-1Beta-511, IL-1RN and TNF-A-308 in the patients with DU compared with control. After stratified by Helicobacter pylori infection, they also could not reach significant differences in this study. No statistically significant differences were observed in DU group compared with control according to combination of the IL-1Beta-511 and IL-1RN genotypes regardless of H. pylori positivity. These findings show that no evidence for the involvement of a proinflammatory polymorphism in the IL-1Beta-511, IL-1RN and TNF-A-308 in the susceptibility to DU in China. PMID- 19804406 TI - A novel Y331X nonsense mutation in TNFRSF1A gene in two unrelated Turkish families with periodic fever syndrome. AB - The autoinflammatory disorders differ in severity, as well as age of onset, duration, and manifestations, but they all share some common features: recurring fever peaks, inflammation of serosal membranes, musculoskeletal involvement, varying types of skin rash, amyloidosis as a sequel of the disease. TRAPS is very rare in Turkish population and we present two unrelated Turkish children with similar clinical phenotypes and laboratory findings related with autoinflammatory disorders and with novel p. Y331X mutation in TNFRSF1A gene. Both of the patients were male and they had recurrent fever without abdominal pain and arthralgia. Full cDNA and exon-intron binding regions of TNFRSF1A, MEFV, MVK, CIAS1 genes were analysed by direct DNA sequencing methods in order to differentiate TRAPS, FMF, HIDS, CINCA/MWS/FCAS respectively. We screened ten exons of TNFRSF1A gene, and detected a heterozygous c.1080C>G nucleotide substitution in exon 10 in both of the unrelated patients, resulting p.Y360X nonsense (protein truncated) mutation. According to classical TNFRSF1A gene nomenclature and the agreement of 30th amino acid as the first one, it is accepted as p.Y331X. It was interesting to determine same mutations in fathers of two patients. In one of the cases, E148Q heterozygous mutation, which is one of the disease-causing mutations of MEFV gene, was detected. No nucleotide substitution was identified in exon and exon-intron splicing regions encoding 396 amino acid of MVK gene in both of the patients. In CIAS1 gene, two different nucleotide substitutions resulting synonymous amino acid mutation were detected in exon 3: c.[732G>A] and c.[786A>G] nucleotide substitutions and compatible p.A242A (according to c.DNA p.A244A) and p.R260R (according to c.DNA p.R262R) synonymous amino acid mutations. These nucleotide substitutions were also detected in parents and were reported to be normal variations in Turkish population. In conclusion, in Turkish patients, with dominantly inherited recurrent fever, TRAPS is a diagnosis worthy of attention and novel mutations have to be reported with phenotype associations. PMID- 19804407 TI - Oct-1 is responsible for the C-33T polymorphism effect in the IL-4 promoter. AB - IL-4 is a pleiotropic immunoregulatory cytokine secreted by Th2 subset of CD4(+) Th cells. Several transcription factors (TFs) have been determined with various degrees of certainty to bind the IL-4 promoter and to regulate its expression in human. To investigate the mechanisms responsible for phenotypic effects of the C 33T IL-4 promoter polymorphism, we performed a search of TFs binding to this promoter locus and discriminating the -33C and -33T alleles. In silico searches suggest few factors bind this region. Using an electromobility shift assay we found that Jurkat T cells contained proteins which specifically interacted with oligonucleotide probes, corresponding to the -33 region. Considerable binding differences between C and T alleles were demonstrated using competitive conditions, the proteins bound predominantly with -33C allele. We found that the transcription factor Oct-1 produced the major shifted complex. The binding of Oct 1 was not improved using activated nuclear extracts; however, we observed increases in other shifted complexes upon cell activation. We suppose that Oct-1 occupancy may compete for binding of activator proteins to closely or overlapped binding sites. Our findings suggest that the interplay between Oct-1 and unknown TFs may be responsible for the C-33T polymorphism effects. PMID- 19804408 TI - NMR structural characterization of HIV-1 virus protein U cytoplasmic domain in the presence of dodecylphosphatidylcholine micelles. AB - The HIV-1 encoded virus protein U (VpU) is required for efficient viral release from human host cells and for induction of CD4 degradation in the endoplasmic reticulum. The cytoplasmic domain of the membrane protein VpU (VpUcyt) is essential for the latter activity. The structure and dynamics of VpUcyt were characterized in the presence of membrane simulating dodecylphosphatidylcholine (DPC) micelles by high-resolution liquid state NMR. VpUcyt is unstructured in aqueous buffer. The addition of DPC micelles induces a well-defined membrane proximal alpha-helix (residues I39-E48) and an additional helical segment (residues L64-R70). A tight loop (L73-V78) is observed close to the C-terminus, whereas the interhelical linker (R49-E63) remains highly flexible. A 3D structure of VpUcyt in the presence of DPC micelles was calculated from a large set of proton-proton distance constraints. The topology of micelle-associated VpUcyt was derived from paramagnetic relaxation enhancement of protein nuclear spins after the introduction of paramagnetic probes into the interior of the micelle or the aqueous buffer. Qualitative analysis of secondary chemical shift and paramagnetic relaxation enhancement data in conjunction with dynamic information from heteronuclear NOEs and structural insight from homonuclear NOE-based distance constraints indicated that micelle-associated VpUcyt retains a substantial degree of structural flexibility. PMID- 19804409 TI - Mammalian glutaminyl cyclases and their isoenzymes have identical enzymatic characteristics. AB - Glutaminyl cyclases (QCs) catalyze the formation of pyroglutamate residues at the N-terminus of several peptides and proteins from plants and animals. Recently, isoenzymes of mammalian QCs have been identified. In order to gain further insight into the biochemical characteristics of isoQCs, the human and murine enzymes were expressed in the secretory pathway of Pichia pastoris. Replacement of the N-terminal signal anchor by an alpha-factor prepropeptide from Saccharomyces cerevisiae resulted in poor secretion of the protein. Insertion of an N-terminal glycosylation site and shortening of the N-terminus improved isoQC secretion 100-fold. A comparison of different recombinant isoQC proteins did not reveal an influence of mutagenic changes on catalytic activity. An initial characterization showed identical modes of substrate conversion of human isoQC and murine isoQC. Both proteins displayed a broad substrate specificity and preference for hydrophobic substrates, similar to the related QC. Likewise, a determination of the zinc content and reactivation of the apo-isoQC revealed equimolar zinc present in QC and isoQC. Far-UV CD spectroscopic analysis of murine QC and isoQC indicated virtually identical structural components. The present investigation provides the first enzymatic characterization of mammalian isoQCs. QC and isoQC represent very similar proteins, which are both present in the secretory pathway of cells. The functions of QCs and isoQC probably complement each other, suggesting a pivotal role of pyroglutamate modification for protein and peptide maturation. PMID- 19804410 TI - Epitope analysis of the rat dipeptidyl peptidase IV monoclonal antibody 6A3 that blocks pericellular fibronectin-mediated cancer cell adhesion. AB - We previously showed that the rat dipeptidyl peptidase IV (rDPP IV) monoclonal antibody (mAb) 6A3 greatly inhibits the pericellular polymeric fibronectin mediated metastatic cancer cell adhesion to rDPP IV. L(311)QWLRRI in rDPP IV has been proposed as the putative fibronectin-binding site. However, the inhibitory mechanism of 6A3 has been elusive. Epitope mapping of 6A3 may help to understand the interaction between fibronectin and rDPP IV. In the present study, we showed that 6A3 species-specifically recognized rDPP IV but inhibited fibronectin/rDPP IV-mediated cell adhesions of various cancer types and species, which was independent of rDPP IV enzymatic activity. The 6A3 epitope was stably exposed in both native and denatured rDPP IV. On the basis of the resolved structures and the species variations in DPP IV sequences, we finely mapped the 6A3 epitope to a surface-exposed Thr331-dependent motif D(329)KTTLVWN, only 11 amino acids away from L(311)QWLRRI on the same plane as the fifth beta-propeller blade. The functionality of 6A3 epitope in rDPP IV was ultimately demonstrated by the ability of 6A3-recognizable fragments to interfere with the inhibitory effect of 6A3 on full-length rDPP IV binding to pericellular polymeric fibronectin. On the basis of structural analysis, and the fact that the preformed fibronectin fragment/rDPP IV complex was co-immunoprecipitated by 6A3 and fixing the rDPP IV structure with paraformaldehyde did not avert the inhibitory effect, the mechanism of 6A3 inhibition may not be the result of complete competition or conformational change. PMID- 19804411 TI - Identification of an atypical insect olfactory receptor subtype highly conserved within noctuids. AB - Olfaction is primarily mediated by the large family of olfactory receptors. Although all insect olfactory receptors share the same structure with seven transmembrane domains, they present poor sequence homologies within and between species. As the only exception, Drosophila melanogaster OR83b and its orthologues define a receptor subtype singularly conserved between insect species. In this article, we report the identification of a new subtype of putative olfactory receptors exceptionally conserved within noctuids, a taxonomic group that includes crop pest insects. Through homology-based molecular cloning, homologues of the previously identified OR18 from Heliothis virescens were identified in the antennae of six noctuid species from various genera, presenting an average of 88% sequence identity. No orthologues were found in genomes available from diverse insect orders and selection pressure analysis revealed that the noctuid OR18s are under purifying selection. The OR18 gene was studied in details in the cotton leafworm, Spodoptera littoralis, where it presented all the characteristic features of an olfactory receptor encoding gene: its expression was restricted to the antennae, with expression in both sexes; its developmental expression pattern was reminiscent of that from other olfactory genes; and in situ hybridization experiments within the antennae revealed that the receptor-expressing cells were closely associated with the olfactory structures, including pheromone- and non pheromone-sensitive structures. Taken together, our data suggest that we have identified a new original subtype of olfactory receptors that are extremely conserved within noctuids and that might fulfil a critical function in male and female noctuid chemosensory neurones. PMID- 19804412 TI - The pro-form of BMP-2 interferes with BMP-2 signalling by competing with BMP-2 for IA receptor binding. AB - Pro-forms of growth factors have received increasing attention since it was shown that they can affect both the maturation and functions of mature growth factors. Here, we assessed the biological function of the pro-form of bone morphogenetic protein-2 (BMP-2), a member of the transforming growth factor beta (TGFbeta)/BMP superfamily. The role of the 263 amino acids of the pro-peptide is currently unclear. In order to obtain an insight into the function of the pro-form (proBMP 2), the ability of proBMP-2 to induce alkaline phosphatase (AP), a marker enzyme for cells differentiating into osteoblasts, was tested. Interestingly, in contrast to mature BMP-2, proBMP-2 did not lead to induction of AP. Instead, proBMP-2 inhibited the induction of AP by BMP-2. This result raised the question of whether proBMP-2 may compete with mature BMP-2 for receptor binding. ProBMP-2 was found to bind to the purified extracellular ligand binding domain (ECD) of BMPR-IA, a high-affinity receptor for mature BMP-2, with a similar affinity as mature BMP-2. Binding of proBMP-2 to BMPR-IA was confirmed in cell culture by cross-linking proBMP-2 to BMPR-IA presented on the cell surface. In contrast to this finding, proBMP-2 did not bind to the ECD of BMPR-II. ProBMP-2 also differed from BMP-2 in its capacity to induce p38 and Smad phosphorylation. The data presented here suggest that the pro-domain of BMP-2 can alter the signalling properties of the growth factor by modulating the ability of the mature part to interact with the receptors. PMID- 19804413 TI - LRRK2 in Parkinson's disease: genetic and clinical studies from patients. AB - Mutations in leucine-rich repeat kinase 2 (LRRK2) (PARK8) are associated with both familial and sporadic forms of Parkinson's disease. Most studies have shown that LRRK2 mutations may explain between 5% and 13% of familial and 1-5% of sporadic Parkinson's disease. Importantly, a common recurrent mutation (G2019S) located in the kinase domain has been reported across most ethnic populations, with the highest prevalence among Ashkenazi Jews and North African Arabs. A recent worldwide meta-analysis pooling data from 24 populations reported a higher occurrence of G2019S in southern than in northern European countries and the penetrance is estimated to be approximately 75% at the age of 79 years. The R1441 'hotspot' amino acid codon residue (G/H/C) in the Ras of complex proteins domain is the second most common site of pathogenic LRRK2 substitutions after G2019S, with most carriers developing symptoms by the age of 75 years. Two polymorphic variants found almost exclusively among Asians (G2385R and R1628P) have been shown to increase the Parkinson's disease risk by approximately two-fold. The mutational event associated with R1628P is more recent, occurring approximately 2500 years ago, compared to estimates of 4000 years for G2385R carriers. LRRK2 mutation carriers generally simulate late onset Parkinson's disease and present with the usual typical clinical features. Genetic testing for G2019S in sporadic late-onset Parkinson's disease can be considered in some situations and may be useful in populations with high carrier status. The identification of asymptomatic mutation and risk variant carriers provides a unique opportunity for recruiting these subjects in potential neuroprotective trials and longitudinal studies to identify biomarkers of neurodegeneration. PMID- 19804414 TI - LRRK2 in Parkinson's disease: in vivo models and approaches for understanding pathogenic roles. AB - The recent discovery of the genetic causes for Parkinson's disease (PD) is fruitful; however, the continuing revelation of PD-related genes is rapidly outpacing the functional characterization of the gene products. Although the discovery of multiple PD-related genes places PD as one of the most complex multigenetic diseases of the brain, it will undoubtedly facilitate the unfolding of a central pathogenic pathway and an understanding of the etiology of PD. Recent findings of pathogenic mutations in leucine-rich repeat kinase 2 (LRRK2) (PARK8) that are linked to the most common familial forms and some sporadic forms of PD provide a unique opportunity to gain insight into the pathogenesis of PD. Despite rapid growth in biochemical, structural and in vitro cell culture studies of LRRK2, the in vivo characterizations of LRRK2 function generally fall short and are largely limited to invertebrates. The investigation of LRRK2 or homologs of LRRK2 in nonmammalian models provides important clues with respect to the cellular functions of LRRK2, but an elucidation of the physiology and pathophysiology of LRRK2 relevant to PD would still depend on mammalian models established by multiple genetic approaches, followed by rigorous examination of the models for pathological process. This minireview summarizes previous studies of genes for ROCO and LRRK2 homologs in slime mold, nematode worms and fruit flies. It also discusses the results obtained from available mouse models of LRRK2 that begin to provide information for understanding LRRK2-mediated pathogenesis in PD. PMID- 19804415 TI - LRRK2 in Parkinson's disease: function in cells and neurodegeneration. AB - The detailed characterization of the function of leucine-rich repeat kinase 2 (LRRK2) may provide insight into the molecular basis of neurodegeneration in Parkinson's disease (PD) because mutations in LRRK2 cause a phenotype with strong overlap to typical late-onset disease and LRRK2 mutations are responsible for significant proportions of PD in some populations. The complexity of large multidomain protein kinases such as LRRK2 challenges traditional functional approaches, although initial studies have successfully defined the basic mechanisms of enzyme activity with respect to the putative effects of pathogenic mutations on kinase activity. The role of LRRK2 in cells remains elusive, with potential function in mitogen-activated protein kinase pathways, protein translation control, programmed cell death pathways and activity in cytoskeleton dynamics. The initial focus on LRRK2-kinase-dependent phenomena places emphasis on the discovery of LRRK2 kinase substrates, although candidate substrates are so far confined to in vitro assays. Here, hypothetical mechanisms for LRRK2-mediated cell death and kinase activation are proposed. As a promising target for neuroprotection strategies in PD, in vitro and in vivo models that accurately demonstrate LRRK2's function relevant to neurodegeneration will aide in the identification of molecules with the highest chance of success in the clinic. PMID- 19804416 TI - LRRK2 in Parkinson's disease: biochemical functions. AB - Leucine-rich repeat kinase 2 (LRRK2) is a large, complex, multidomain protein containing kinase and GTPase enzymatic activities and multiple protein-protein interaction domains. Mutations linked to autosomal dominant forms of Parkinson's disease result in amino acid changes throughout the protein and alterations in both its enzymatic properties and interactions. The best characterized mutation to date, G2019S, leads to increased kinase activity, and mutations in the GTPase domain, such as R1441C and R1441G, have also been reported to influence kinase activity. Therefore, an examination of LRRK2's properties as a kinase is important for understanding the mechanisms underlying the disorder and has the potential to lead to therapeutics. These findings also suggest that there may be complex interplay between the functional domains of LRRK2. Here, we review LRRK2's biochemical functions based on structural and kinetic studies of the enzymatic domains, its potential substrates and the role of its interactions. Despite the field's embryonic understanding of the true relevance of these substrates and interactions, initial studies are providing clues with respect to its pathophysiological functions. Together, these findings should increase our understanding of mechanisms underlying Parkinson's disease and place LRRK2 as a unique molecular target for effective therapeutic development. PMID- 19804417 TI - LRRK2 in Parkinson's disease: building an understanding of disease etiology. PMID- 19804418 TI - Pitfalls of Melan-A staining. PMID- 19804419 TI - Haemodynamic effects of patent foramen ovale and atrial septal defect closure: a comparison during percutaneous shunt closure. AB - SUMMARY OBJECTIVES: We investigated the haemodynamic effect of percutaneous closure of an intra-atrial shunt, using non-invasive finger pressure measurements. BACKGROUND: Percutaneous closure of both patent foramen ovale (PFO) and atrial septal defect (ASD) is widely practised. Currently no data are available on short-term haemodynamic changes induced by closure. METHODS: Twenty five consecutive patients (mean age 49 +/- 17 years, 10 men) who underwent a percutaneous closure of a PFO (n = 15) or ASD (n = 10) were included in this study. During the procedure blood pressure and heart rate (HR) were monitored continuously with a Finometer. Changes in systolic, mean, and diastolic pressure, stroke volume (SV), cardiac output (CO) and total peripheral resistance (TPR) were computed from the pressure registrations using Modelflow methodology. RESULTS: Baseline characteristics were similar for the PFO and ASD patients. After PFO closure none of the haemodynamic parameters changed significantly. After ASD closure the systolic, mean, and diastolic pressures increased 7.1 +/- 5.4 (P = 0.003), 3.8 +/- 3.5 (P = 0.007) and 2.0 +/- 3.0 mmHg (P = ns) respectively. HR decreased 5.1 +/- 5.3 beats per minute (P = 0.01). SV, CO and TPR increased 8.5 +/- 6.4 ml (13.5%; P = 0.002), 0.21 +/- 0.45 l min(-1) (5.6%; P = ns) and 0.02 +/- 0.14 dynes (4.1%; P = ns) respectively. The changes in SV differ between the PFO and ASD patients (P = 0.009). CONCLUSIONS: Using non invasive finger pressure measurements, we found that SV, mean and systolic blood pressure increased immediately after percutaneous closure of an ASD in adults, whereas the percutaneous PFO closure had no effect on haemodynamic characteristics. PMID- 19804420 TI - Accuracy of implant impressions with different impression coping types and shapes. AB - BACKGROUND: Accurate recording of implant location is required so that definitive restorations are properly supported and do not place additional stresses on the implants. Movement of impression copings inside the impression material using an open-tray or close-tray impression technique during clinical and laboratory phases may cause inaccuracy in transferring the three-dimensional spatial orientation of implants intraorally to the definitive cast. Consequently, the restoration may require corrective procedures. AIM: This in vitro study compared the accuracy of two different impression techniques with two different impression coping shapes using polyether impression material to obtain precise definitive casts. MATERIALS AND METHODS: Two reference acrylic resin models (Technovits 4000, Heraeus Kulzer GmbH & Co., Wehrheim, Germany) with five internal connection implants having different shapes of impression copings (Implantium [Dentium, Seoul, South Korea] and Replace Select [Nobel Biocare AB, Goteborg, Sweden]) were fabricated. Twenty medium-consistency polyether impressions of these models were made with square and conical impression copings of each system using open-tray and close-tray techniques. Matching implant replicas were screwed into the impression copings in the impressions. Impressions were poured with type IV stone, and the positional accuracy of the implant replica heads in x-, y-, and z axes (represented in [Deltar]) and also rotational displacement (DeltaTheta) were evaluated using a coordinate measuring machine (Mistral, DEA Brown&Sharpe, Grugliasco, Italy). These measurements (linear and rotational displacements) were compared with the measurements calculated on the reference resin models that served as control, and data were analyzed with a two-way analysis of variance at alpha = 0.05. RESULTS: Less inaccuracy occurred in less retentive shape impression copings (Replace Select) compared with the more retentive one (Implantium) (p(r) < .001 and p(Theta) < .001), but there was no significant difference between direct and indirect impression techniques (p(r) and p(Theta) > .05). CONCLUSION: The impression coping shape had more impact on impression inaccuracy than impression technique did. Understanding of the magnitude and variability of distortion when employing certain impression-making methods and impression coping shapes helps the clinician to select a better implant component and impression technique. PMID- 19804421 TI - Suppression of gastric cancer dissemination by ephrin-B1-derived peptide. AB - Interaction of the Eph family of receptor protein tyrosine kinases and their ligands, ephrin family members, induces bidirectional signaling through cell-cell contacts. High expression of B-type ephrin is associated with high invasion potential of tumors, and we previously observed that signaling through the C terminus of ephrin-B1 mediates the migration and invasion of cells, and is involved in the promotion of carcinomatous peritonitis in vivo. Here we show that the intracellular introduction of a synthetic peptide derived from ephrin-B1 C terminus blocks ephrin-B1 mediated signaling in scirrhous gastric cancer cells. Treatment of cancer cells with a fusion peptide consisting of HIV-TAT and amino acids 331-346 of ephrin-B1 (PTD-EFNB1-C) suppressed the activation of RhoA, mediated by the association of ephrin-B1 with an adaptor protein Dishevelled, and also inhibited extracellular secretion of metalloproteinase. Moreover, injection of PTD-EFNB1-C peptide into the peritoneal cavity of nude mice suppressed carcinomatous peritonitis of intraperitoneally transplanted scirrhous gastric cancer cells. These results indicate the possible application of ephrin-B1 C terminal peptide to develop novel protein therapy for scirrhous gastric carcinoma, especially in the stage of tumor progression, including peritoneal dissemination. PMID- 19804422 TI - Effects of Src inhibitors on cell growth and epidermal growth factor receptor and MET signaling in gefitinib-resistant non-small cell lung cancer cells with acquired MET amplification. AB - The efficacy of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors such as gefitinib and erlotinib in non-small cell lung cancer (NSCLC) is often limited by the emergence of drug resistance conferred either by a secondary T790M mutation of EGFR or by acquired amplification of the MET gene. We now show that the extent of activation of the tyrosine kinase Src is markedly increased in gefitinib-resistant NSCLC (HCC827 GR) cells with MET amplification compared with that in the gefitinib-sensitive parental (HCC827) cells. In contrast, the extent of Src activation did not differ between gefitinib-resistant NSCLC (PC9/ZD) cells harboring the T790M mutation of EGFR and the corresponding gefitinib-sensitive parental (PC9) cells. This activation of Src in HCC827 GR cells was largely abolished by the MET-TKI PHA-665752 but was only partially inhibited by gefitinib, suggesting that Src activation is more dependent on MET signaling than on EGFR signaling in gefitinib-resistant NSCLC cells with MET amplification. Src inhibitors blocked Akt and Erk signaling pathways, resulting in both suppression of cell growth and induction of apoptosis, in HCC827 GR cells as effectively as did the combination of gefitinib and PHA-665752. Furthermore, Src inhibitor dasatinib inhibited tumor growth in HCC827 GR xenografts to a significantly greater extent than did treatment with gefitinib alone. These results provide a rationale for clinical targeting of Src in gefitinib-resistant NSCLC with MET amplification. PMID- 19804423 TI - Tumor-host histopathologic variables, stromal myofibroblasts and risk score, are significantly associated with recurrent disease in tongue cancer. AB - Margin status, a major prognostic parameter in oral cancer, was analyzed vis-a vis the histopathologic parameters of risk scores and stromal myofibroblasts. Specimens of tongue carcinoma (n = 50) were submitted to a risk score assignment consisting of the worst pattern of invasion, lymphocytic infiltration, and perineural invasion. Frequency of stromal myofibroblasts (alpha-smooth muscle actin stain) was assessed. A triple immunostaining assay with E-cadherin, Ki-67 and alpha-smooth muscle actin was used to identify carcinoma cells undergoing epithelial-mesenchymal transition. Margins were considered 'clean' if the tumor was >or=5 mm away from them. Patients or =8 significantly increased the risk of death. On multivariate analysis only the surgical Gleason score had an independent prognostic role with regard to overall survival (P = 0.01; odds ratio 2.82, 95% confidence interval 1.2-6.4) and cancer-specific survival (P < 0.001; 8.6, 2.5-28.8). CONCLUSIONS: In this series, overall and cancer-specific survival rates were comparable to those reported for surgically treated cT3 prostate cancers. The lack of need for external urinary diversion during the entire follow-up significantly contributed to the patients' quality of life. PMID- 19804425 TI - Type 2 diabetes but not metabolic syndrome is associated with an increased risk of lower urinary tract symptoms and erectile dysfunction in men aged <45 years. AB - OBJECTIVE: To investigate the association of type 2 diabetes mellitus (T2DM) and metabolic syndrome with lower urinary tract symptoms (LUTS) and erectile dysfunction (ED) in Taiwanese men aged <45 years. PATIENTS, SUBJECTS AND METHODS: Voiding and erectile function in 226 men with T2DM, at one diabetes clinic, and 183 healthy men with normal fasting blood glucose levels, were compared. Participants were evaluated using the International Prostate Symptom Score (IPSS), the five-item version of the International Index of Erectile Function questionnaire (IIEF-5), and measurements of flow rate and postvoid residual urine volume. The association of metabolic syndrome with LUTS and ED was also evaluated. RESULTS: The mean (sd, range) age of the patients was 38.9 (6.1, 20 45) years and the mean duration of diabetes was 2.8 (3.1, 0.5-20) years. Compared with controls, men with T2DM had a significantly mean (sd) higher IPSS, of 6.1 (5.8) vs 4.1 (4.6) (P < 0.001), an increased of odds ratio (95% confidence interval) of having moderate to severe LUTS of 1.78 (1.12-2.84) (P = 0.01), greater voiding volume of 376 (177) vs 326 (102) mL (P = 0.04), a worse IIEF-5 score of 17.3 (6.4) vs 20.0 (3.8) (P < 0.001), an increased of odds ratio of having moderate to severe ED of 3.5 (2.1-5.8) (P < 0.001) but a similar maximum flow rate and postvoid residual. The IIEF-5 score was negatively correlated with the IPSS (P < 0.0001, coefficient = -0.23, 0.35-0.11) and glycosylated haemoglobin (P = 0.02, coefficient = -0.14, 0.26-0.01). In all, 156 (69%) patients met the criteria for metabolic syndrome. The mean age, duration of diabetes, glycosylated haemoglobin, IPSS, voided volume, maximum urinary flow rate and IIEF-5 score were similar between patients with and without metabolic syndrome. CONCLUSIONS: Men with T2DM and aged <45 years had more LUTS but a similar bladder emptying function than the controls. ED was highly prevalent and was associated with the severity of LUTS. Metabolic syndrome did not aggravate the severity of LUTS, emptying function or ED in the early stage of DM. PMID- 19804426 TI - Laparoscopic transperitoneal radical prostatectomy in renal transplant recipients: a review of the literature. AB - OBJECTIVES: To report our experience with laparoscopic radical prostatectomy (LRP) for the treatment of localized prostate carcinoma in two renal transplant recipients and a review of the literature. PATIENTS AND METHODS: We retrospectively identified all patients who had undergone LRP for clinically localized prostate cancer between 2002 and 2008 at our institution (n = 1150). Of these patients, two were renal transplant recipients (one with donor renal transplant cadaver and the other with prior transplantectomy). We reviewed all available clinicopathological data and the scientific literature. RESULTS: The two patients underwent successful LRP with no major complications. The mean (range) operative time was 200 (180-220) min with a mean estimated blood loss of 300 (200-400) mL. There were no changes in renal graft function as measured by serum creatinine level. At pathology, the surgical margins were negative and disease was organ-confined in each case. The two patients tolerated the procedure well and had a mean (range) hospital stay of 3.5 (3-4) days. CONCLUSIONS: The data from our two patients suggest that LRP, as an accepted minimally invasive treatment for a middle-aged man with organ-confined prostate cancer, is a technically feasible and safe treatment of localized prostate cancer in renal transplant recipients. PMID- 19804427 TI - Robotic-assisted radical prostatectomy: a review of current outcomes. AB - With the widespread diffusion of the screening for prostate cancer, the disease has been diagnosed more commonly in the organ-confined stage, and in younger and healthier men. For these patients, radical prostatectomy (RP) is still the standard treatment. In an effort to decrease the morbidity associated with open RP, minimally invasive approaches have been described, including robotic-assisted RP (RALP). Almost one decade after the introduction of RALP, large and mature series have now been reported. We reviewed the outcomes of the largest series of RALP published recently. We searched Medline for reports published between 2006 and 2009, to identify articles describing intraoperative data, surgical complications, oncological outcomes, continence and potency rates after RALP. Relevant articles were selected and the outcomes evaluated. PMID- 19804428 TI - Trends in epidemiology and treatment of upper urinary tract tumours in the Netherlands 1995-2005: an analysis of PALGA, the Dutch national histopathology registry. AB - OBJECTIVE: To evaluate changes in incidence, distribution of stage and grade as well as surgical treatment of upper urinary tract (UUT) tumours in the Netherlands from 1995 to 2005. PATIENTS AND METHODS: The PALGA registry, a nationwide network and registry of pathology encompassing all hospitals in the Netherlands, was used as primary data source. Pathology reports of all primary surgical procedures or biopsies without further surgical treatment within the next year, of cancer of the renal pelvis or ureter during the period 1995-2005, were included. The number of surgically treated UUT tumours per year, type of treatment and tumour characteristics were recorded. RESULTS: The population consisted of 2321 (67%) men and 1145 (33%) women with a mean age of 68.6 years. The distribution according to side was similar (left 44.1%, right 41.5%), bilateral tumours were rare (0.6%) and most tumours were in the renal pelvis (51.3%). Both the incidence and the incidence rate per 100 000 person-years increased during the study period (P < 0.001). Most urothelial cancers were grade 2 (40.9%) or 3 (41.2%) and stage Ta (30.6%), T1 (18.1%) or T3 (22.8%). There was an increase in grade 3 (P = 0.003) and muscle-invasive (P = 0.003) tumours in men only. Nephroureterectomy was performed in 41.3% of the cases and there was an increasing trend to endoscopic surgery (P = 0.019), although the absolute number was low. CONCLUSION: The incidence of surgically treated UUT tumours increased, with a significant trend towards more advanced disease in men. Most tumours were treated by nephroureterectomy or nephrectomy, although there was an increasing trend to endoscopic surgery. PMID- 19804429 TI - An audit of intensive care unit recyclable waste. AB - There is little known about recyclable intensive care unit waste. We tested the hypotheses that the intensive care unit produces a small proportion (< 10%) of hospital waste, that much waste (> 30%) is recyclable and that there is little (< 10%) cross-contamination of non-infectious with infectious waste. For seven consecutive days in an Australian 10-bedded intensive care unit, we prospectively sorted all waste. The total intensive care unit waste for the week was 540 kg, representing 5% of hospital waste. Of the 401 kg of intensive care unit general waste, recyclables were 230 kg (57%; 95% CI 53-61%), mainly plastics, cardboard and paper. There were 0.4 kg of infectious cross-contamination in the 401 kg of general waste. Intensive care unit waste was a small proportion of all hospital waste. However, there was minimal infectious waste cross-contamination and almost 60% of intensive care unit general waste could be recycled with appropriate safeguards, education and training. PMID- 19804430 TI - Cost-effectiveness of temsirolimus for first line treatment of advanced renal cell carcinoma. AB - OBJECTIVES: To estimate the cost-effectiveness of temsirolimus compared to interferon-alpha for first line treatment of patients with advanced, poor prognosis renal cell carcinoma, from the perspective of the UK National Health Service. METHODS: A decision-analytic model was developed to estimate the cost effectiveness of temsirolimus. The clinical effectiveness of temsirolimus compared with interferon-alpha and the utility values (using EQ-5D tariffs) were taken from a recent phase III randomized clinical trial. Cost data were obtained from published literature and based on current UK practice. The effect of parameter uncertainty on cost-effectiveness was explored through extensive one way and probabilistic sensitivity analyses. RESULTS: Compared to interferon alpha, temsirolimus treatment resulted in an incremental cost per QALY gained of pound94,632; based on an estimated mean gain of 0.24 quality-adjusted life years (QALYs) per patient, at a mean additional cost of pound22,331 (inflated to 2007/8). The cost per QALY for patient subgroups ranged from pound74,369 to pound154,752. The probability that temsirolimus is cost-effective compared to interferon-alpha at a willingness to pay threshold of pound30,000 per QALY for all patient groups is expected to be close to zero. The cost per QALY was sensitive to the clinical effectiveness parameters, health state utilities, drug costs and the cost of administration of temsirolimus. CONCLUSIONS: Temsirolimus has been shown to be clinically effective compared to interferon-alpha offering additional health benefits, however, with a cost per QALY in excess of pound90,000, it may not be regarded as a cost-effective use of resources in some health care settings. PMID- 19804431 TI - Cost-effectiveness of sorafenib for second-line treatment of advanced renal cell carcinoma. AB - OBJECTIVES: To estimate the cost-effectiveness of sorafenib (Nexavar, Bayer, Leverkusen, Germany) versus best supportive care (BSC) for second-line treatment of advanced renal cell carcinoma from the perspective of the UK National Health Service. METHODS: A decision analytic model was developed to estimate the cost effectiveness of sorafenib. The clinical effectiveness of sorafenib versus BSC was taken from a recent randomized phase III trial. Utility values were taken from a phase II trial of sunitinib, using EQ-5D tariffs. Cost data were obtained from published literature and were based on current UK practice. The effect of parameter uncertainty on cost-effectiveness was explored through extensive one way and probabilistic sensitivity analyses. RESULTS: Compared to BSC, sorafenib treatment resulted in an incremental cost per quality-adjusted life year (QALY) gained of pound75,398, based on an estimated mean gain of 0.27 QALYs per patient, at a mean additional cost of pound20,063 (inflated to 2007/2008). The probability that sorafenib is cost-effective compared to BSC at a willingness to pay threshold of pound30,000 per QALY is 0.0%. In sensitivity analysis, estimates of cost per QALY were sensitive to changes in the clinical effectiveness parameters, and to health state utilities and drug costs. CONCLUSIONS: Sorafenib has been shown to be clinically effective compared to BSC, offering additional health benefits; however, with a cost per QALY in excess of pound70,000, it may not be regarded as a cost-effective use of resources in some health-care settings. PMID- 19804432 TI - Reliability and validity of a Chinese version's health-related quality of life questionnaire for hepatitis B patients. AB - OBJECTIVES: To culturally adapt a Chinese version of the Hepatitis Quality of Life Questionnaire (HQLQ) and assess its suitability for use in Chinese-speaking hepatitis B virus (HBV patients in Singapore. STUDY: Reliability was assessed using Cronbach's alpha coefficients and intra-class correlation coefficients. Item-to-scale correlation was assessed using Spearman's rank correlations (rho) between scale scores and their constituent items. Convergent and divergent construct validities were tested in three and two a priori hypotheses, respectively, and the correlations were assessed using Spearman's rank correlation coefficients. RESULTS: When tested in 134 HBV patients, the test retest reliability was supported with all scales showing acceptable correlation coefficients (i.e., alpha>0.7). Item-to-scale correlations were good with most items highly correlated with their hypothesized scales. Convergent and divergent construct validities were supported by the hypothesized correlations between the HQLQ and the EQ-5D domains. CONCLUSIONS: The culturally adapted questionnaire has good validity and reliability for use in Singapore. PMID- 19804433 TI - In or out? Income losses in health state valuations: a review. AB - BACKGROUND: In 1996 the Washington Panel controversially recommended valuing productivity costs (PC) in terms of quality-adjusted life years. The Panel's assumption that respondents in health state valuation (HSV) exercises take income losses into account could not be countered since there was no evidence regarding what people consider in HSV exercises. If they do consider income losses and if this changes HSVs, then all economic evaluations that have included PC in the numerator may have double-counted these costs. Alternatively, if respondents do not consider income losses then all past economic evaluations that have not included PC in the numerator have failed to account for sizeable societal costs. OBJECTIVES: Through a review we aim to recapture the debate surrounding the appropriate method for including PC in health economic evaluations, to identify empirical evidence addressing the assumptions of the Panel, and recommend a future research agenda. METHODS: Through a review we identify, outline, and critically appraise the existing empirical studies that attempt to address whether respondents include income effects in HSV exercises. RESULTS AND CONCLUSION: Seven empirical studies were identified. Overall, it seems that not explicitly mentioning the inclusion of income will induce a minority of respondents to include these effects and this appears not to influence results. More empirical work is needed, using generic instruments, larger samples, and using the interview method of administration. PMID- 19804434 TI - Impact of the 1997 Canadian guidelines on the conduct of Canadian-based economic evaluations in the published literature. AB - OBJECTIVE: To assess the impact of the 1997 Canadian guidelines on the methods and presentation of economic evaluations conducted from a Canadian perspective in the published literature. METHODS: A systematic literature review was conducted to identify health technology economic evaluations conducted from a Canadian perspective published in peer-reviewed journals between 2001 and 2006. To investigate the impact of the 1997 Canadian Coordinating Office of Health Technology Assessment guidelines, each included study was assessed against 17 of the 25 recommendations. RESULTS: Of the 153 included studies, a base set of 9 methodological standards, as outlined by the 1997 guidelines, were followed by over 50% of the studies including: indications, outcomes for cost utility analysis, outcomes for cost benefit analysis, discounting future cost and outcomes, cost identification and valuation, evaluating uncertainty and disclosing funding relationships. Main divergences from the guidelines were found for analytic technique (38%), study perspective (23%), source of preferences (8%), equity (7%), and cost measurement (24%). CONCLUSION: The current assessment has shown that the 1997 Canadian guidelines have set a minimum methodological standard within the community of "doers" conducting economic analyses from a Canadian perspective. Although there was divergence from some of the recommendations, the majority were reflected as changes in the 2006 Canadian guidelines. PMID- 19804435 TI - Crossing borders: factors affecting differences in cost-effectiveness of smoking cessation interventions between European countries. AB - OBJECTIVES: Many different factors affect the transferability of cost effectiveness results between countries. The objective is to quantify the impact of nine potential causes of variation in cost-effectiveness of pharmacological smoking cessation therapies (SCTs) between The Netherlands (reference case), Germany, Sweden, UK, Belgium, and France. METHODS: The life-time benefits of smoking cessation were calculated using the Benefits of Smoking Cessation on Outcomes model, following a cohort of smokers making an unaided quit attempt, or using nicotine replacement therapy (NRT), bupropion, or varenicline. We investigated the impact of between-country differences in nine factors demography, smoking prevalence, mortality, epidemiology and costs of smoking related diseases, resource use and unit costs of SCTs, utility weights and discount rates-on the incremental net monetary benefit (INMB), using a willingness-to-pay (WTP) of euro20,000 per quality adjusted life year (QALY). RESULTS: The INMB of 1000 quit attempts with NRT versus unaided, varies from euro0.39 million (Germany) to euro1.47 million (France). The differences between the countries were primarily due to differences in discount rates, causing the INMB to change between -65% to +62%, incidence and mortality rates (epidemiology) of smoking-related diseases (-43% to +35%) and utility weights. Impact also depended on the WTP for a QALY and time horizon: at a low WTP or a short time horizon, the resource use and unit costs of SCTs had the highest impact on INMB. CONCLUSIONS: Although all INMBs were positive, there were significant differences across countries. These were primarily related to choice of discount rate and epidemiology of diseases. PMID- 19804436 TI - Cost-effectiveness of 21 alternative cervical cancer screening strategies. AB - OBJECTIVES: The aim of this study is to assess the cost-effectiveness of 21 alternative cervical cancer screening (CCS) strategies. METHODS: A cohort simulation model was developed to determine from a health systems perspective the cost-effectiveness of the 21 alternative CCS strategies that incorporated combinations of Papanicolaou's smear test (PAP), liquid-based cytology (LBC) or human papillomavirus deoxyribonucleic acid (HPV-DNA) testing. The model was calibrated to categorize total costs into four budgetary authorities: testing, physician, inpatient, and outpatient services. Within each category, alternative screening strategies were contrasted in terms of their cost impacts and the percent change calculated within each category. Epidemiologic data and costs were derived from administrative health databases. Estimates of test characteristics and quality-adjusted life years (QALYs) were derived from available literature. RESULTS: Three-year screening with PAP and HPV-DNA triage testing for women older than 30 years of age (3-year PAP+HPV+PAP-age) is less costly and more effective saving $16,078 per additional QALY gained. Although there was an associated net cost decrease of 4.2% driven by a reduction in testing and physician costs of 22.1% and 18.6%, respectively, there is a cost increase of 0.8% and 27.7% in inpatient and outpatient services, respectively. CONCLUSION: There is economic evidence to support adopting 3-year PAP+HPV+PAP-age. Budgetary resources can potentially be shifted from testing and physician services to fund the additional resource requirements for inpatient and outpatient services. PMID- 19804437 TI - Use of existing patient-reported outcome (PRO) instruments and their modification: the ISPOR Good Research Practices for Evaluating and Documenting Content Validity for the Use of Existing Instruments and Their Modification PRO Task Force Report. AB - BACKGROUND: Patient-reported outcome (PRO) instruments are used to evaluate the effect of medical products on how patients feel or function. This article presents the results of an ISPOR task force convened to address good clinical research practices for the use of existing or modified PRO instruments to support medical product labeling claims. The focus of the article is on content validity, with specific reference to existing or modified PRO instruments, because of the importance of content validity in selecting or modifying an existing PRO instrument and the lack of consensus in the research community regarding best practices for establishing and documenting this measurement property. METHODS: Topics addressed in the article include: definition and general description of content validity; PRO concept identification as the important first step in establishing content validity; instrument identification and the initial review process; key issues in qualitative methodology; and potential threats to content validity, with three case examples used to illustrate types of threats and how they might be resolved. A table of steps used to identify and evaluate an existing PRO instrument is provided, and figures are used to illustrate the meaning of content validity in relationship to instrument development and evaluation. RESULTS & RECOMMENDATIONS: Four important threats to content validity are identified: unclear conceptual match between the PRO instrument and the intended claim, lack of direct patient input into PRO item content from the target population in which the claim is desired, no evidence that the most relevant and important item content is contained in the instrument, and lack of documentation to support modifications to the PRO instrument. In some cases, careful review of the threats to content validity in a specific application may be reduced through additional well documented qualitative studies that specifically address the issue of concern. CONCLUSION: Published evidence of the content validity of a PRO instrument for an intended application is often limited. Such evidence is, however, important to evaluating the adequacy of a PRO instrument for the intended application. This article provides an overview of key issues involved in assessing and documenting content validity as it relates to using existing instruments in the drug approval process. PMID- 19804438 TI - Anaphylaxis to Patent Blue V. II. A unique IgE-mediated reaction. AB - BACKGROUND: Patent Blue V (PBV) is injected in order to map sentinel nodes during cancer staging procedures. Anaphylactic reactions, allegedly IgE antibody mediated, have been reported. The aim of the study was to explore the immunological mechanism of anaphylaxis to PBV. METHODS: PBV allergen threshold basophil sensitivity, CD-sens, was performed on cells from nine patients diagnosed as having had adverse reactions to PBV. The mechanisms of the CD-sens were studied by immunological and immuno-chemical methods. RESULTS: Five of the nine patients had a positive CD-sens to PBV which was completely eliminated by washing the cells in phosphate buffered saline before allergen challenge. However, the positive CD-sens was completely reconstituted by incubating the cells in plasma or serum of that patient or the other PBV-anaphylactic patients for 15 min at room temperature. In some patients the factor mediating CD-sens was completely or partially destroyed by heating at +56 degrees C for 30 min or being exposed to the low pH used for elution from anti-Ig columns. A 1000-fold excess of monoclonal IgE blocked the reconstitution by approximately 50%. CONCLUSION: Anaphylactic reactions to PBV are mediated by IgE antibodies giving a classical CD-sens reaction. However, the allergenic configuration seems to constitute a structure completely dependent on PBV, as a hapten, linked to a, so far, unknown carrier that seems to be unique for patients having experienced a PBV-induced reaction. Further studies are needed to characterize the postulated carrier. PMID- 19804439 TI - Passive smoking is a major determinant of exhaled nitric oxide levels in allergic asthmatic children. AB - BACKGROUND: Fraction of exhaled nitric oxide (FeNO) is considered, by some authors, to be a treatment follow-up parameter in allergic asthmatics. However, factors such as active smoking can influence NO production and must be taken into account in the interpretation of FeNO values. In children, the evidence in favour of an impact of passive smoking (PS) on FeNO values is controversial. The aim of this study was to evaluate the impact of chronic PS on FeNO in allergic asthmatic children. METHODS: Seventy nontreated allergic asthmatic children over 5 years of age, exposed and unexposed to PS, underwent measurement of FeNO, spirometry, and allergic tests (skin prick tests, total and specific serum IgE, and blood eosinophilia). Children were considered to be exposed to PS when at least 1 cigarette per day was declared to be smoked at home. RESULTS: Geometric mean FeNO value in 22 children exposed to PS was 26.3 +/- 1.5 ppb vs 56.3 +/- 1.7 ppb in 48 children unexposed (P < 0.001). After adjustment for age, blood eosinophilia, allergic sensitizations, total IgE, dust mite sensitization and asthma severity, multivariate analysis showed that PS exposure was negatively associated with FeNO values (P = 0.0001) and was the primary determinant of FeNO variations. CONCLUSION: Passive smoking lowers FeNO, and might be a major determinant of FeNO levels in nontreated allergic asthmatic children. PMID- 19804440 TI - Efficacy of omalizumab in delayed pressure urticaria: a case report. PMID- 19804441 TI - Steroid sparing effects of intranasal corticosteroids in asthma and allergic rhinitis. AB - BACKGROUND: Treating allergic rhinitis may have a downstream anti-inflammatory effect on the lower airways. We conducted a dose ranging study in asthma and persistent allergic rhinitis to evaluate if intranasal corticosteroids exhibit a sparing effect on the dose of inhaled corticosteroid. METHODS: Twenty five participants were randomized to receive two weeks of 100 microg/day (Low dose) or 500 microg/day (High dose) of inhaled fluticasone propionate both with intranasal placebo; or inhaled fluticasone 100 microg/day with intranasal fluticasone 200 microg/day (Combined) in a double-blind cross-over fashion. RESULTS: Low dose fluticasone produced a shift of 1.20 doubling-dilutions (95% CI, 0.63, 1.77); Combined fluticasone, 1.79 doubling-dilutions (95% CI, 0.77, 2.80) and high dose fluticasone, 2.01 doubling-dilutions (95% CI, 1.42, 2.61) in methacholine PC(20) from respective baselines. There was a significant difference between high and low doses: 0.82 doubling dilutions (95%CI, 0.12, 1.50) but not between combined and low dose 0.58 doubling dilutions (95% CI, -0.78, 1.95). Combined treatment alone produced improvements in peak nasal inspiratory flow (P < 0.001), rhinitis quality of life (P = 0.004) and nasal NO (P = 0.01); reduced blood eosinophil count (P = 0.03), and serum eosinophil cationic protein (P = 0.02). All treatments significantly improved tidal NO, FEV(1) and asthma quality of life. CONCLUSIONS: High-dose fluticasone was superior to low dose fluticasone for methacholine PC20, demonstrating room for further improvement. Combined treatment was not significantly different from low dose fluticasone and we could not demonstrate a steroid sparing effect on methacholine PC20. Combined treatment alone produced improvements in upper airway outcomes and suppressed systemic inflammation but not adrenal function. PMID- 19804442 TI - Allergic contact stomatitis to cinnamon in chewing gum mistaken as facial angioedema. PMID- 19804443 TI - The allergen specificity of the late asthmatic reaction. AB - BACKGROUND: Allergen inhalation challenge in asthma may induce both early (EAR) and late (LAR) asthmatic reactions. The EAR is IgE and mast cell dependent. The mechanism of the LAR is less well defined and we have hypothesized may be allergen dependent. The aim of this study was to investigate the allergen specificity of the LAR to allergen inhalation in asthma. METHODS: In a randomized, double-blind, crossover design six asthmatic volunteers with dual sensitization to house dust mite (HDM) allergen and grass pollen (GP) allergen underwent inhalation allergen challenge with these separate allergens on two occasions separated by 14 days. Lung function changes were followed for 8-h postchallenge. Bronchial reactivity (histamine PC(20)) and airway inflammation, assessed by induced sputum differential cell count, were measured 24-h pre and postallergen challenge. The allergen inhalation challenges were matched to achieve the same magnitude of EAR. RESULTS: Despite comparable group mean EAR percent falls in FEV(1) (25.8% following GP and 28.0% following HDM (P = 0.917), the LAR was statistically greater on the HDM challenge day (13.0%vs 22.8% [P = 0.046]) and was associated with a significant airway eosinophil recruitment (mean (SD) of 5.4 (4.8)% to 22.1 (18.2)% (P = 0.028) that was not evident on the GP allergen challenge day. CONCLUSIONS: These findings identify the allergen specificity of the LAR and indicate that factors independent of IgE contribute to the LAR. Such findings have relevance both to the understanding of the allergen induced airway responses in asthma and the need for homogeneity in inhaled allergen challenge studies in asthma. PMID- 19804444 TI - Time-series nasal epithelial transcriptomics during natural pollen exposure in healthy subjects and allergic patients. AB - BACKGROUND: The role of epithelium has recently awakened interest in the studies of type I hypersensitivity. OBJECTIVE: We analysed the nasal transcriptomics epithelial response to natural birch pollen exposure in a time series manner. METHODS: Human nasal epithelial cell swabs were collected from birch pollen allergic patients and healthy controls in winter season. In addition, four specimens at weekly intervals were collected from the same subjects during natural birch pollen exposure in spring and transcriptomic analyses were performed. RESULTS: The nasal epithelium of healthy subjects responded vigorously to allergen exposure. The immune response was a dominating category of this response. Notably, the healthy subjects did not display any clinical symptoms regardless of this response detected by transcriptomic analysis. Concomitantly, the epithelium of allergic subjects responded also, but with a different set of responders. In allergic patients the regulation of dyneins, the molecular motors of intracellular transport dominated. This further supports our previous hypothesis that the birch pollen exposure results in an active uptake of allergen into the epithelium only in allergic subjects but not in healthy controls. CONCLUSION: We showed that birch pollen allergen causes a defence response in healthy subjects, but not in allergic subjects. Instead, allergic patients actively transport pollen allergen through the epithelium to tissue mast cells. Our study showed that new hypotheses can arise from the application of discovery driven methodologies. To understand complex multifactorial diseases, such as type I hypersensitivity, this kind of hypotheses might be worth further analyses. PMID- 19804445 TI - Temperature modulated histamine-itch in lesional and nonlesional skin in atopic eczema - a combined psychophysical and neuroimaging study. AB - BACKGROUND: Itch is the major symptom of many allergic diseases; yet it is still difficult to measure objectively. The aim of this study was to use an evaluated itch stimulus model in lesional (LS) and nonlesional (NLS) atopic eczema (AE) skin and to characterize cerebral responses using functional magnetic resonance imaging (fMRI). METHODS: Thermal modulation was performed on a histamine stimulus in randomized order on LS or NLS in rapid alternating order from 32 degrees C (warm) to 25 degrees C (cold). Subjective itch ratings were recorded. Additionally, fMRI measurements were used to analyze the cerebral processing (n = 13). Healthy skin (HS) of age-matched volunteers served as control (n = 9). RESULTS: Mean VAS itch intensity was significantly (P < 0.0001) higher during the relative cold [55.2 +/- 8.3% (LS); 48.6 +/- 8.2% (NLS)] compared to the relative warm blocks [36.0 +/- 7.3% (LS); 33.7 +/- 7.6% (NLS)]. Compared to HS, the itch response was delayed in LS and NLS. Itch intensity was perceived highest in LS, followed by NLS and HS. For NLS, fMRI revealed at the beginning of the itch provocation a cerebral deactivation pattern in itch processing structures (thalamus, prefrontal, cingulate, insular, somatosensory and motor cortex). During the course of stimulation, the cerebral deactivation was reduced with time and instead an activation of the basal ganglia occurred. In contrast LS showed an activation instead of deactivation pattern already at the beginning of the stimulation in the above mentioned structures. CONCLUSIONS: Moderate short-term temperature modulation led to a reproducible, significant enhancement of histamine-induced itch with the strongest effect in LS. The differences in itch perception and itch kinetics between healthy volunteers and NLS in patients point towards an ongoing central inhibitory activity patients with AE, especially at the beginning of the itch provocation. PMID- 19804446 TI - Basophil activation in two cases of hydrochlorothiazide-induced noncardiogenic pulmonary edema. PMID- 19804447 TI - Osteopontin is expressed and functional in human eosinophils. AB - BACKGROUND: Eosinophils are critically involved in allergic inflammation and tissue remodeling. Osteopontin (OPN) is a glycoprotein molecule which exhibits pro-fibrogenic and pro-angiogenic properties and has recently also been implicated in allergic diseases. In this study, we investigated the expression and function of OPN in human eosinophils. METHODS: Osteopontin mRNA (RT-PCR) and protein (immunofluorescence) expression in peripheral blood eosinophils from atopic human subjects were evaluated. Soluble OPN release was determined in resting and activated eosinophils. The contribution of OPN to eosinophil-induced angiogenesis was determined using the chick embryo chorio- allantoic membrane (CAM) assay and OPN-induced eosinophil chemotaxis was determined (ChemoTx System microplate wells). Finally, OPN expression in bronchoalveolar lavage (BAL) fluids from mild asthmatic and normal control subjects was determined. RESULTS: Osteopontin is expressed in human eosinophils and is increased following GM-CSF and IL-5 activation. Eosinophil-derived OPN contributes to eosinophil-induced angiogenesis. Recombinant OPN promotes eosinophil chemotaxis in vitro and this effect is mediated by alpha(4)beta(1) integrin binding. Soluble OPN is increased in the bronchoalveolar lavage fluid from mild asthmatic subjects and correlates with eosinophil counts. CONCLUSIONS: We therefore conclude that OPN is likely to contribute to the process of angiogenesis observed in the airways in asthma. PMID- 19804448 TI - Steroid dependency despite omalizumab treatment of ABPA in cystic fibrosis. PMID- 19804449 TI - Matrix metalloproteinases-7, -8, -9 and TIMP-1 in the follow-up of diisocyanate induced asthma. AB - BACKGROUND: Diisocyanate-induced asthma (DIA) is known to be associated with poor prognosis. We wished to clarify if matrix metalloproteinases (MMP)-7, -8 or -9 or tissue inhibitor of matrix metalloproteinases (TIMP-1) are associated with the functional or inflammatory outcome in DIA patients. METHODS: This is a longitudinal study where 17 patients with DIA diagnosed by a specific challenge test to diisocyanates were monitored. Exposure to diisocyanates was terminated seven (mean) months before the challenge test. The studies included spirometry, histamine challenge test and bronchoscopy. MMP-7, MMP-8, TIMP-1 [Enzyme-linked immunosorbent assay (ELISA)- and immunofluorometric assay-methods], MMP-9 (ELISA and zymography), interferon-gamma, tumour necrosis factor-alpha, interleukin-6, 8, -15, -17, CXCL-5/ENA-78, monocyte chemoattractant protein-1 and macrophage inhibitory factor (MIF) (ELISA) were assayed from bronchoalveolar lavage (BAL) fluid. Inhaled steroid therapy was initiated after the examinations, which were repeated at 6 months and at 3 years during the treatment. The results were compared with those of 15 healthy controls. RESULTS: Inhaled steroid medication increased BAL levels of MMP-9 and MMP-9/TIMP-1 and decreased MMP-7 and MMP-7/TIMP 1. The increase in MMP-9 levels was associated with a decline in the TH-2 type inflammation. CONCLUSIONS: Our data suggest that reduced TH-2 type inflammation in DIA after inhaled steroid medication is reflected as elevated MMP-9 and MMP 9/TIMP-1 levels in BAL. MIF may be the inducer of MMP-9. This might point to some protective role for MMP-9 in DIA. PMID- 19804450 TI - Grass transcutaneous immunotherapy in children with seasonal rhinoconjunctivitis. PMID- 19804451 TI - IRF4 polymorphism rs872071 and risk of Hodgkin lymphoma. AB - The reciprocal familial risk between chronic lymphocytic leukaemia (CLL) and Hodgkin lymphoma (HL) suggests genetic variants with pleiotropic effects may influence the risk of both CLL and HL. We have recently shown that the IRF4 variant rs872071 influences CLL risk. To examine if rs872071 genotype is associated with HL risk we genotyped two case-control series (totalling, 529 and 2192, respectively). This analysis provides evidence that IRF4 rs872071 influences HL risk; Odds Ratio = 1.21 (95% confidence interval: 1.05-1.39, P = 0.009) and highlights the importance of inherited variation in B-cell developmental genes in the development of HL. PMID- 19804452 TI - Epstein-Barr virus negative large B-cell lymphoma during long term immunomodulatory therapy for T-cell large granular lymphocytic leukaemia. PMID- 19804453 TI - Mutations in the fifth immunoglobulin-like domain of kit are common and potentially sensitive to imatinib mesylate in feline mast cell tumours. AB - The purpose of the current study was to investigate the mutation status of KIT in feline mast cell tumours (MCTs) and to examine the effects of tyrosine kinase inhibition on the phosphorylation of mutant kit in vitro and in clinical cases of cats. Sequence analysis of KIT identified mutations in 42/62 MCTs (67.7%). The vast majority of the mutations were distributed in exons 8 and 9, both of which encode the fifth immunoglobulin-like domain (IgD) of kit. All five types of kit with a mutation in the fifth IgD were then expressed in 293 cells and examined for phosphorylation status. The mutant kit proteins showed ligand-independent phosphorylation. The tyrosine kinase inhibitor imatinib mesylate suppressed the phosphorylation of these mutant kit proteins in transfectant cells. In a clinical study of 10 cats with MCTs, beneficial response to imatinib mesylate was observed in 7/8 cats that had a mutation in the fifth IgD of kit in tumour cells. Mutations in the fifth IgD of kit thus appear to be common and potentially sensitive to imatinib mesylate in feline MCTs. These data provide an in vivo model for paediatric mastocytosis where mutations in the fifth IgD of kit also occur. PMID- 19804454 TI - Combination of KIT gene silencing and tocopherol succinate may offer improved therapeutic approaches for human mastocytosis. AB - Gain-of-function mutations of kit tyrosine kinase receptor are associated with mastocytosis. Two subclones of the HMC1 mast leukaemia cell line were used; both express an identical KIT allele-specific regulatory type mutation (V560G), but differ in that one also expresses an enzymatic site type mutation (D816V) that confers on them resistance to imatinib mesylate tyrosine kinase inhibitor. In both cell lines, proliferation was suppressed and apoptosis induced by the combination of KIT gene silencing and alpha-tocopherol succinate (alpha-TOS), a derivate of alpha-tocopherol, also known as vitamin E. Furthermore, HMC1 cells with decreased kit levels by KIT silencing, failed to form tumours when xenotransplanted into immunocompromised mice and the animals were treated systemically with alpha-TOS. Targeting kit in the presence of alpha-TOS represents a new approach against proliferation of human mast leukaemia cell lines. PMID- 19804455 TI - Failure of three novel regimens to improve outcome for patients with relapsed or refractory acute myeloid leukaemia: a report from the Eastern Cooperative Oncology Group. AB - The treatment of relapsed acute myeloid leukaemia (AML) remains unsatisfactory. We conducted a phase II randomized trial where patients received intermediate dose cytarabine for 4 d followed by gemtuzumab ozogamicin on day 5 (Arm A), or combined with liposomal daunorubicin for 3 d (Arm B), or cytarabine given for 5 d combined with cyclophosphamide for 3 d and topotecan by continuous infusion for 5 d (Arm C). Eligible patients had primary refractory AML, a first relapse after a remission of <1 year, or a second or greater relapse. The primary objective of this trial was attainment of a conventional complete remission (CR) or a CR without platelet recovery (CRp) in at least 40% of patients. The CR/CRp rates for the 82 eligible patients were 3/26 (12%) in Arm A, 2/29 (7%) in Arm B, and 1/27 (4%) in Arm C. No patients who had relapsed within 6 months of initial CR or who had suffered multiple relapses responded. More than 95% of patients subsequently died of AML. No unexpected toxicities were encountered. We conclude that none of these three regimens were effective enough in the treatment of high-risk relapsed or refractory AML to warrant further study. This trial was registered at http://www.clinicaltrials.gov as #NCT00005962. PMID- 19804456 TI - Alcohol consumption in homicide victims in the city of Sao Paulo. AB - AIMS: To assess the association between alcohol use and victimization by homicide in individuals autopsied at the Institute of Legal Medicine in Sao Paulo, Brazil. DESIGN: Cross-sectional study. SETTING: Excessive consumption of alcohol is a serious public health issue and a major factor in triggering violent situations, which suggests a strong association between alcohol ingestion and becoming a victim of homicide. PARTICIPANTS: Data from 2042 victims of homicides in 2005 were obtained from medical examiner reports. MEASUREMENTS: The victim's gender, age, ethnicity and blood alcohol concentration (BAC) were collected. The method of death and homicide circumstances, as well as the date, time and place of death were also studied. FINDINGS: Alcohol was detected in blood samples of 43% of the victims, and mean BAC levels were 1.55 +/- 0.86 g/l. The prevalence of positive BAC levels was higher among men (44.1%) than women (26.6%), P < 0.01. Firearms caused most of the deaths (78.6%), and alcohol consumption was greater among victims of homicide by sharp weapons (P < 0.01). A greater proportion of victims with positive BAC were killed at weekends compared to weekdays (56.4 and 38.5%, respectively; P < 0.01), and the correlation between homicide rates and the average BAC for the central area of the city was positive (r(s) = 0.90; P < 0.01). CONCLUSIONS: These results highlight alcohol as a contributing factor for homicide victimization in the greatest urban center in South America, supporting public strategies and future research aiming to prevent homicides and violence related to alcohol consumption. PMID- 19804457 TI - Smoker sensitivity to retail tobacco displays and quitting: a cohort study. AB - AIMS: To assess whether sensitivity to point of sale (POS) cigarette displays influences quitting behaviour. DESIGN: Prospective cohort study. SETTING: Victoria, Australia. PARTICIPANTS: A total of 222 adult smokers were surveyed at baseline in 2006 and followed-up 18 months later. MEASUREMENTS: Baseline sensitivity to POS displays, which included the frequency of 'noticing displays', 'impulse purchasing behaviour' and 'deciding on brand based on POS displays'; smoking status at follow-up. FINDINGS: At follow-up, 17.0% were no longer smokers. After adjusting for covariates, compared to those with low POS display sensitivity, smokers who had a medium or high level of sensitivity to POS displays were significantly less likely to have quit at follow-up [odds ratio (OR) = 0.32, 95% confidence interval (CI) = 0.14-0.74; OR = 0.27, 95% CI = 0.08 0.91, respectively]. CONCLUSIONS: The presence of cigarette pack displays in stores may make it more difficult for smokers to quit smoking successfully. PMID- 19804458 TI - The challenge of external validity in policy-relevant systematic reviews: a case study from the field of substance misuse. AB - AIM: To critically evaluate the methods utilized in the conduct of a systematic review in the field of substance misuse. DESIGN: Participant-observation in the review process, semi-structured interviews with review team members and management and structured observation of the process of guidance development. SETTING: An 'arm's-length' government body. PARTICIPANTS: Review team members, management and the committee responsible for producing evidence-based guidance for policy and practice. MEASUREMENTS: Data from interviews and (participant )observation were reflected upon critically in order to increase understanding of the systematic review process. FINDINGS: The application of systematic review methods produced an evidence base that did not inform the development of guidance to the extent that it could have done: (i) an emphasis upon internal research validity produced an evidence base with an emphasis on short-term interventions at the level of the individual; (ii) criteria for appraising the external validity of studies were not developed sufficiently; and (iii) the systematic review of evidence and development of guidance are strongly reliant upon the judgement of reviewers and committee members. CONCLUSIONS: Prioritizing internal validity in a systematic review risks producing an evidence base that is not informed adequately by the wider determinants of health and which does not give sufficient consideration to external validity. The use of appropriate methods requires that commissioners of systematic reviews are clear at the outset how the review is proposed to be utilized. Review methods such as meta-ethnography and realist synthesis could contribute to making the frameworks within which judgements are made more explicit. PMID- 19804459 TI - Influence of smoking cues in movies on craving among smokers. AB - AIMS: Research has shown that smoking-related cues are important triggers for craving. The objective of the present study was to test whether smoking cues in movies also function as triggers to evoke craving. To accomplish this, we conducted a pilot study in which we examined smokers' reactivity to smoking cues from a particular movie in a common cue-reactivity paradigm using pictures. In the main study, we tested whether smokers who are confronted with smoking characters in a movie segment have a greater desire to smoke than smokers confronted with non-smoking characters. DESIGN: Using an experimental design, participants were assigned randomly to one of two movie conditions (smoking versus non-smoking characters). SETTING: In a laboratory, that reflected a naturalistic setting, participants watched a 41-minute movie segment. PARTICIPANTS: A total of 65 young adults who smoked on a daily basis participated in the experiment. MEASUREMENTS: Craving was assessed before and after watching the movie. FINDINGS: The pilot study revealed that pictures of smoking characters had strong effects on craving. However, when smokers actually watched a movie segment, no differences in craving were found between those who watched smoking characters and those who watched non-smoking characters. This finding was not affected by baseline craving, the time of the last cigarette smoked and daily smoking habits. CONCLUSIONS: No effect of smoking cues in movies on craving was found, in contrast with research supporting the cue-craving link. Thus, if replicated, this might indicate that smoking cues in such contexts do not affect smokers' desire to smoke as expected. PMID- 19804460 TI - Extending drug ethno-epidemiology using agent-based modelling. AB - AIMS: To show how the inclusion of agent-based modelling improved the integration of ethno-epidemiological data in a study of psychostimulant use and related harms among young Australians. METHODS: Agent-based modelling, ethnographic fieldwork, in-depth interviews and epidemiological surveys. SETTING: Melbourne, Perth and Sydney, Australia. PARTICIPANTS: Club drug users in Melbourne, recreational drug users in Perth and street-based injecting drug users in Sydney. Participants were aged 18-30 years and reported monthly or more frequent psychostimulant use. FINDINGS: Agent-based modelling provided a specific focus for structured discussion about integrating ethnographic and epidemiological methods and data. The modelling process was underpinned by collective and incremental design principles, and produced 'SimAmph', a data-driven model of social and environmental agents and the relationships between them. Using SimAmph, we were able to test the probable impact of ecstasy pill-testing on the prevalence of harms--a potentially important tool for policy development. The study also navigated a range of challenges, including the need to manage epistemological differences, changes in the collective design process and modelling focus, the differences between injecting and non-injecting samples and concerns over the dissemination of modelling outcomes. CONCLUSIONS: Agent-based modelling was used to integrate ethno-epidemiological data on psychostimulant use, and to test the probable impact of a specific intervention on the prevalence of drug-related harms. It also established a framework for collaboration between research disciplines that emphasizes the synthesis of diverse data types in order to generate new knowledge relevant to the reduction of drug-related harms. PMID- 19804461 TI - Content of ecstasy in the Netherlands: 1993-2008. AB - AIMS: The present paper outlines the results of analyses carried out on the content of tablets sold as ecstasy, collected in the Netherlands by the Drugs Information Monitoring System (DIMS) from January 1993 to December 2008. METHODS: During a period of 16 years, the DIMS analysed the content of 33 006 tablets sold as ecstasy that were handed in by numerous individual (potential) substance users. The DIMS results were compared with the results from various seized tablets to determine whether the DIMS is a monitor of the ecstasy consumer market. RESULTS: The DIMS system appears to be a market monitor that gives an accurate reflection of what is actually available on the hidden Dutch ecstasy market. During 16 years of monitoring, the purity [tablets containing only 3,4 methylenedioxymethamphetamine (MDMA)] was lowest around 1997. During this time period many tablets contained other substances in addition to or instead of MDMA [e.g. 3,4-methylene-dioxyamphetamine (MDA), 3,4-methylene-dioxyethylamphetamine (MDEA) and N-methyl-a-(1,3-benzodixol-5-yl)-2-butamine (MBDB), amphetamine and caffeine]. From 1998 to 2008, the number of high-dose tablets (> or =106 mg MDMA per tablet) gradually increased. The same holds true for the proportion of tablets that contained only MDMA, reaching the highest levels in 2000 and 2004. After 2004, the purity of ecstasy tablets decreased again, caused mainly by a growing proportion of tablets containing meta-chlorophenylpiperazine (mCPP). CONCLUSIONS: The DIMS results provide valuable qualitative information on the content of ecstasy tablets in the Netherlands, and its changes throughout the years. Moreover, the results were used for national and international risk assessments and important warning and prevention activities. PMID- 19804462 TI - Do Delta9-tetrahydrocannabinol concentrations indicate recent use in chronic cannabis users? AB - AIMS: To quantify blood Delta(9)-tetrahydrocannabinol (THC) concentrations in chronic cannabis users over 7 days of continuous monitored abstinence. PARTICIPANTS: Twenty-five frequent, long-term cannabis users resided on a secure clinical research unit at the US National Institute on Drug Abuse under continuous medical surveillance to prevent cannabis self-administration. MEASUREMENTS: Whole blood cannabinoid concentrations were determined by two dimensional gas chromatography-mass spectrometry. FINDINGS: Nine chronic users (36%) had no measurable THC during 7 days of cannabis abstinence; 16 had at least one positive THC > or =0.25 ng/ml, but not necessarily on the first day. On day 7, 6 full days after entering the unit, six participants still displayed detectable THC concentrations [mean +/- standard deviation (SD), 0.3 +/- 0.7 ng/ml] and all 25 had measurable carboxy-metabolite (6.2 +/- 8.8 ng/ml). The highest observed THC concentrations on admission (day 1) and day 7 were 7.0 and 3.0 ng/ml, respectively. Interestingly, five participants, all female, had THC positive whole blood specimens over all 7 days. Body mass index did not correlate with time until the last THC-positive specimen (n = 16; r = -0.2; P = 0.445). CONCLUSIONS: Substantial whole blood THC concentrations persist multiple days after drug discontinuation in heavy chronic cannabis users. It is currently unknown whether neurocognitive impairment occurs with low blood THC concentrations, and whether return to normal performance, as documented previously following extended cannabis abstinence, is accompanied by the removal of residual THC in brain. These findings also may impact on the implementation of per se limits in driving under the influence of drugs legislation. PMID- 19804463 TI - Estimating population attributable risk for hepatitis C seroconversion in injecting drug users in Australia: implications for prevention policy and planning. AB - OBJECTIVE: To determine risk factors and estimate their population-level contribution to hepatitis C virus (HCV) burden. METHODS: Established and potentially modifiable risk factors were estimated using partial population attributable risk (PAR(p)) in a cohort of new injecting drug users (IDUs) in Sydney, Australia. RESULTS: A total of 204 hepatitis C seronegative IDUs were recruited through street-based outreach, methadone clinics and needle and syringe programmes (NSPs) and followed-up at 3-6-monthly intervals. A total of 61 HCV seroconversions were observed during the follow-up [overall incidence rate of 45.8 per 100 person-years (95% confidence interval: 35.6-58.8)]. Overall, five potentially modifiable risk factors (sharing needles/syringes, sharing other injecting equipment, assisted injecting, frequency of injection and not being in drug treatment) accounted for approximately 50% of HCV cases observed. CONCLUSION: While sharing needles/syringes or other injecting equipment were associated most strongly with increased risk of HCV infection, the PAR(p) associated with these behaviours was relatively modest (12%) because they are relatively low-prevalence behaviours. Our analyses suggest that more HCV infection could be avoided by changing more common, but less strongly associated behaviours such as assisted injecting or daily injecting. Results suggest that to have a very substantial effect on HCV, a range of risk factors need modifying. The most efficient use of scarce resources in reducing HCV infections will require complex balancing between the PAR for a given risk factor(s), the efficacy of interventions to actually modify the risk factor, and the cost of these interventions. PMID- 19804464 TI - Alcohol and hypertension: gender differences in dose-response relationships determined through systematic review and meta-analysis. AB - AIMS: To analyze the dose-response relationship between average daily alcohol consumption and the risk of hypertension via systematic review and meta-analysis. DESIGN: A computer-assisted search was completed for 10 databases, followed by hand searches of relevant articles. Only studies with longitudinal design, quantitative measurement of alcohol consumption and biological measurement of outcome were included. Dose-response relationships were assessed by determining the best-fitting model via first- and second-degree fractional polynomials. Various tests for heterogeneity and publication bias were conducted. FINDINGS: A total of 12 cohort studies were identified from the literature from the United States, Japan and Korea. A linear dose-response relationship with a relative risk of 1.57 at 50 g pure alcohol per day and 2.47 at 100 g per day was seen for men. Among women, the meta-analysis indicated a more modest protective effect than reported previously: a significant protective effect was reported for consumption at or below about 5 g per day, after which a linear dose-response relationship was found with a relative risk of 1.81 at 50 g per day and of 2.81 at an average daily consumption of 100 g pure alcohol per day. Among men, Asian populations had higher risks than non-Asian populations. CONCLUSIONS: The risk for hypertension increases linearly with alcohol consumption, so limiting alcohol intake should be advised for both men and women. PMID- 19804465 TI - The alcohol industry and public interest science. AB - AIMS: This report argues that the growing involvement of the alcohol industry in scientific research needs to be acknowledged and addressed. It suggests a set of principles to guide ethical decision-making in the future. METHODS: We review relevant issues with regard to relationships between the alcohol industry and the international academic community, especially alcohol research scientists. The guiding principles proposed are modelled after expert committee statements, and describe the responsibilities of governmental agencies, the alcohol industry, journal editors and the academic community. These are followed by recommendations designed to inform individuals and institutions about current 'best practices' that are consistent with the principles. FINDINGS AND CONCLUSIONS: Growing evidence from the tobacco, pharmaceutical and medical fields suggests that financial interests of researchers may compromise their professional judgement and lead to research results that are biased in favour of commercial interests. It is recommended that the integrity of alcohol science is best served if all financial relationships with the alcoholic beverage industry are avoided. In cases where research funding, consulting, writing assignments and other activities are initiated, institutions, individuals and the alcoholic beverage industry itself are urged to follow appropriate guidelines that will increase the transparency and ethicality of such relationships. PMID- 19804466 TI - Systematic review: efficacy and safety of pancreatic enzyme supplements for exocrine pancreatic insufficiency. AB - BACKGROUND: Pancreatic enzyme supplements are standard therapy for fat malabsorption in patients with exocrine pancreatic insufficiency. The FDA determined that published data are insufficient to support the efficacy and safety of these agents. AIM: To determine if pancreatic enzyme supplements are: (i) superior to placebo for treating fat malabsorption and (ii) superior to other supplements based on randomized cross-over trials. METHODS: A computer-assisted search of MEDLINE and EMBASE was performed to identify relevant studies. Data extraction on study design, improvement in coefficient of fat absorption, diarrhoea and adverse events using prespecified forms. RESULTS: A total of 12 manuscripts met inclusion criteria. Most studies (10/12) compared pancreatic enzyme supplements that used different delivery systems, while using similar quantities of enzymes. These studies found no consistent difference in fat malabsorption or gastrointestinal symptoms between different active treatments. Two small placebo-controlled trials (n = 65 patients) demonstrate that pancreatic enzyme supplements are superior to placebo for fat absorption. Data are inadequate to determine if pancreatic enzyme supplements lead to weight gain or improvement in diarrhoea. CONCLUSIONS: Based on data from randomized cross-over trials, pancreatic enzyme supplements appear to improve fat malabsorption. No specific branded product or specific delivery system is superior for treatment of fat malabsorption in patients with exocrine pancreatic insufficiency. PMID- 19804467 TI - Acute pyelonephritis with renal abscesses and acute renal failure after salmonella infection. AB - Urinary tract infections, renal abscess formation and acute renal failure (ARF) after salmonella infection are rarely reported in children. We present a previously healthy teenager who developed ARF with renal abscess formation after salmonella infection, in whom we believe that acute salmonella pyelonephritis was the main causative factor for ARF and not dehydration, shock or rhabdomyolysis, which have already been described in the literature. With prolonged antibiotic treatment and adequate hydration, the boy's condition improved, but chronic kidney disease was unfortunately inevitable. CONCLUSION: Salmonella pyelonephritis has, according to our knowledge, not yet been described to be the main causative factor of ARF in previously healthy children, as was the case in our patient. Long-term antibiotic treatment of at least 6 weeks is probably a must in such patients, even though chronic kidney disease could not have been prevented. PMID- 19804468 TI - Opinion of Belgian neurologists on antiepileptic drug treatment in 2006: Belgian study on epilepsy treatment (BESET-2). AB - OBJECTIVES: (i) To describe the medical treatment of epilepsy in Belgium in 2006, (ii) to detect the presence or absence of consensus in epilepsy treatment and (iii) to analyze the evolution of the neurologists' opinion between 2003 and 2006. MATERIALS AND METHODS: In December 2006, 100 neurologists were interviewed with a structured questionnaire, based on ordinal four-point scales. The questionnaire contained questions on treatment choices in adult patients with epilepsy. The results of this survey were compared with results of a previous one done in 2003. RESULTS: Initial monotherapy was the preferred treatment strategy. Valproate was first choice in idiopathic generalized epilepsy. Carbamazepine and oxcarbazepine were first choice in focal epilepsy with partial seizures. Valproate was also first choice in focal epilepsy with secondarily generalized seizures. New antiepileptic drugs were recommended in second line. However, in special treatment situations, they were considered first-line, e.g. lamotrigine in case of women in childbearing age. In comparison with 2003, there was a trend of using earlier the new antiepileptic drugs. CONCLUSIONS: In end 2006, carbamazepine, valproate and oxcarbazepine were considered to be first choice drugs, whereas other newer drugs, like lamotrigine, levetiracetam and topiramate were predominantly prescribed in second line. PMID- 19804469 TI - Hyperdense middle cerebral artery sign is an ominous prognostic marker despite optimal workflow. AB - OBJECTIVES: To evaluate the association between the hyperdense middle cerebral artery sign (HMCAS) and the functional outcome on one hand, and different predictors such as the National Institutes of Health Stroke Scale (NIHSS), infarct size, ASPECTS Score, intracerebral hemorrhage, and mortality on the other hand. MATERIAL AND METHODS: Retrospective analysis of 120 patients with MCA stroke treated with intravenous thrombolysis. We tested the association between HMCAS and NIHSS, infarct volume, ASPECTS, outcome, level of consciousness, different recorded time intervals, and the day/time of admission. RESULTS: Seventy-four percentage of patients treated with thrombolysis developed cerebral infarction. All patients with HMCAS (n = 39) sustained infarction and only 31% showed favorable outcome compared with 62% and 60%, respectively among patients without HMCAS (P < 0.001 and P = 0.002). There was statistically significant association between functional outcome and HMCAS (P = 0.002), infarct volume, NIHSS, and ASPECTS (P < 0.001). The time to treatment was 12 min shorter in patients who developed infarction (P = 0.037). Independent predictors for outcome were NIHSS and the occurrence of cerebral infarction on computed tomography for the whole study population, and infarct volume for patients who sustained cerebral infarction. CONCLUSIONS: Despite optimal workflow, patients with HMCAS showed poor outcome after intravenous thrombolysis. The results emphasize the urgent need for more effective revascularization therapies and neuroprotective treatment in this subgroup of stroke patients. PMID- 19804470 TI - Less protease-resistant PrP in a patient with sporadic CJD treated with intraventricular pentosan polysulphate. AB - Treatment with intraventricular pentosan polysulphate (PPS) might be beneficial in patients with Creutzfeldt-Jakob disease. We report a 68-year-old woman with sporadic Creutzfeldt-Jakob disease who received continuous intraventricular PPS infusion (1-120 microg/kg/day) for 17 months starting 10 months after the onset of clinical symptoms. Treatment with PPS was well tolerated but was associated with a minor, transient intraventricular hemorrhage and a non-progressive collection of subdural fluid. The patient's overall survival time was well above the mean time expected for the illness but still within the normal range. Post mortem examination revealed that the level of abnormal protease-resistant prion protein in the brain was markedly decreased compared with levels in brains without PPS treatment. These findings suggest that intraventricular PPS infusion might modify the accumulation of abnormal prion proteins in the brains of patients with sporadic Creutzfeldt-Jakob disease. PMID- 19804471 TI - Mitral valve prolapse and abnormalities of haemostasis in children and adolescents with migraine with aura and other idiopathic headaches: a pilot study. AB - OBJECTIVE: To investigate the prevalence of mitral valve prolapse (MVP) and abnormalities of haemostasis in children and adolescents with migraine with aura (MA) compared with peers affected by other idiopathic headaches. MATERIALS AND METHODS: We recruited 20 MA patients (10 men and 10 women; age range 8-17 years) and 20 sex- and age-matched subjects with other idiopathic headaches. Both groups underwent colour Doppler transthoracic echocardiography to detect MVP and the following laboratory work-up: plasma prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen, protein C, protein S, homocysteine, lupus anticoagulant, von Willebrand factor (vWF) ristocetin cofactor activity, immunoglobulins (Ig) G and M anticardiolipin antibodies (aCL). Factor V Leiden, factor II and methylenetetrahydrofolate reductase were investigated (we did not test the entire genes, but screened for specific point mutations). RESULTS: The prevalence of MVP was significantly higher in the MA subjects than in the patients affected by other idiopathic headaches (40% vs 10%; P < 0.05). Moreover, the MA patients showed a higher rate of above-normal IgM aCL titres (45% vs 10%; P < 0.05). Finally, in the group of patients with MVP we found a higher prevalence of aCL in those with MA compared with those affected by other idiopathic headaches. CONCLUSIONS: A proportion, at least, of the MA patients showed a more complex phenotype characterized by MVP and/or positive aCL titres. The pathogenetic role of these associations is obscure and larger studies are needed to confirm the usefulness of echocardiographic and laboratory investigations in this area and to identify possible new treatment approaches that might be explored in this group of MA patients. PMID- 19804472 TI - Leptin and the metabolic syndrome in patients with myotonic dystrophy type 1. AB - OBJECTIVES: To evaluate serum leptin concentration and its relation to metabolic syndrome (MSy) in non-diabetic patients with myotonic dystrophy type 1 (DM1). MATERIALS AND METHODS: This study included 34 DM1 patients, and the same number of healthy subjects matched for age, sex and body mass index (BMI). RESULTS: DM1 patients had increased BMI and insulin resistance, and increased leptin and insulin concentrations, but the other features of MSy such as diabetes, glucose intolerance and hypertension were not detected in DM1 patients. Serum leptin levels were higher in patients with DM1 than in healthy controls (8.5 +/- 6.6 ng/ml vs 3.6 +/- 2.9 ng/ml in men, and 13.9 +/- 10.0 ng/ml vs 10.9 +/- 6.9 ng/ml in women, respectively). In DM1 patients, leptin levels correlated with BMI, fasting insulin and insulin resistance (HOMA) (P < 0.01). CONCLUSIONS: The leptin overproduction correlated with insulin resistance in DM1 patients but the significance of this finding remains unclear. PMID- 19804473 TI - In vitro neurotoxic properties and excitatory aminoacids concentration in the cerebrospinal fluid of amyotrophic lateral sclerosis patients. Relationship with the degree of certainty of disease diagnoses. AB - OBJECTIVE: To determine glutamate and aspartate levels in the cerebrospinal fluid (CSF) in patients with sporadic amyotrophic lateral sclerosis (SALS) grouped according to El Escorial diagnostic criteria, and to perform an in vitro assessment of the neurotoxicity of the CSF in murine cortical neurons. METHODS: SALS patients were sorted according to El Escorial diagnostic criteria. Glutamate and aspartate were measured in the CSF using high performance liquid chromatography. Cultured cortical neuron viability was determined after exposure to CSF for 24 h. RESULTS: Glutamate levels were elevated in 28 out of the 29 patients with definite, probable or possible SALS. There were no differences in glutamate concentrations when the three clinical forms of the disease were compared; neither there were significant variation across disease duration and clinical presentation. In agreement with previous reports, we concluded that CSF SALS-induced in vitro neurotoxicity is mediated by ionotropic glutamate receptors. We found no relationship between the degree of in vitro neurotoxicity and glutamate concentration in the CSF. CONCLUSIONS: Glutamate but not aspartate CSF levels may contribute to ALS pathogenesis. However, glutamate levels may not influence the degree of diagnosis certainty or lesion extension. PMID- 19804474 TI - Endothelial nitric oxide synthase Glu298Asp, 4b/a, and -786T>C gene polymorphisms and the risk of ischemic stroke. AB - BACKGROUND AND PURPOSE: Endothelial nitric oxide synthase (eNOS) gene polymorphisms were associated with reduced NO production, and were evaluated as risk factors for ischemic stroke (IS). We investigated the association between eNOS gene -786T>C (promoter), 27-bp repeat 4b/4a (intron 4), and Glu298Asp (exon 7) polymorphisms with IS in 329 IS patients and 444 controls. MATERIALS AND METHODS: Glu298Asp and -786T>C genotyping was done by PCR-RFLP, 4b/4a was assessed by PCR-ASA. The contribution of eNOS polymorphisms to IS was analyzed by haplotype and multivariate regression analysis. RESULTS: Higher frequency of 298Asp allele was seen in IS patients (P = 1.2 x 10(-10)), which remained independently associated with IS on multivariate analysis after controlling for traditional cerebrovascular risk factors. Allele and genotype distribution of 4b/4a and -786T>C polymorphisms were comparable between patient and controls. Significantly higher prevalence of 298Asp/4b/-786T and 298Asp/4b/-786C haplotypes were seen in IS cases, thus conferring a disease susceptibility nature to these haplotypes. Multivariate regression analysis confirmed the association of 298Asp/4b/-786T and 298Asp/4b/-786C haplotypes, and in addition identified 298Asp/4a/-786T haplotype to be independently associated with IS, after controlling for traditional cerebrovascular risk factors. CONCLUSIONS: Genetic variation at the eNOS locus represent genetic risk factor for increased susceptibility to IS. PMID- 19804475 TI - Drug-induced confusional states: the usual suspects? AB - BACKGROUND: Acute confusional state (ACS) is a frequent reason for hospital admission. This study examines retrospectively the frequency by which individual drugs were found responsible for ACS. RESULTS: Drug-induced ACS was found in 65 (18.8%) of 346 hospital admissions for acute confusion. The most frequent causative substances were dopaminergic drugs in Parkinsonian patients (24.2%), diuretics (15.1%), tricyclic or tetracyclic antidepressants (13.6%) and benzodiazepines (13.6%). Almost half of the patients were demented, and in one third of these, dementia had not been diagnosed hitherto. CONCLUSION: The data suggest that diuretics by way of causing hyponatraemia are as relevant a cause of ACS as dopaminergic or anticholinergic substances. PMID- 19804476 TI - Outcome prediction in traumatic brain injury: comparison of neurological status, CT findings, and blood levels of S100B and GFAP. AB - OBJECTIVE: To investigate the predictive value of early serum levels of S100B and glial fibrillary acidic protein (GFAP) in traumatic brain injury. METHODS: Sixty patients admitted within 24 h of trauma were included. Neurological status on admission (Glasgow Coma Scale), initial cranial computed tomography (CCT) studies (Marshall Computed Tomographic Classification), and outcome after 6 months (Glasgow Outcome Scale) were evaluated. S100B and GFAP levels were determined on admission and 24 h after trauma. RESULTS: Blood levels of S100B and GFAP were elevated following head trauma and quantitatively reflected the severity of trauma. S100B levels after 24 h and on admission were of higher predictive value than CCT findings or clinical examination. GFAP, but not S100B levels rapidly declined after trauma. CONCLUSIONS: Blood levels of S100B and GFAP indicate the severity of brain damage and are correlated with neurological prognosis after trauma. Both methods can yield additional prognostic information if combined with clinical and CCT findings. PMID- 19804477 TI - ApolipoproteinE epsilon 4 allele is not associated with disease course and severity in multiple sclerosis. AB - BACKGROUND: No study has assessed the association between apolipoproteinE (APOE) and multiple sclerosis (MS) forms grouped by also taking into account cognitive performance. AIMS OF THE STUDY: To assess the relationship between APOE and disease course, particularly focusing on benign MS (BMS), defined as also including cognitive preservation. METHODS: In 173 consecutive patients, we assessed the association between APOE and MS course and severity. RESULTS: Twenty nine APOE-epsilon4 carriers were identified. The epsilon4 allele was not associated with BMS. Moreover, it was associated neither with other disease courses nor with the time to reach disability milestones and secondary progression. CONCLUSION: Although plausible, the association between APOE and MS course (particularly with BMS defined by including cognitive preservation) and disease severity remains controversial. PMID- 19804478 TI - Tactile direction discrimination and vibration detection in diabetic neuropathy. AB - OBJECTIVE: To evaluate the clinical usefulness of quantitative testing of tactile direction discrimination (TDD) in patients with diabetic neuropathy. MATERIALS AND METHODS: TDD and vibration detection were examined on the dorsum of the feet in 43 patients with type 1 diabetes mellitus and clinical signs and symptoms indicating mild neuropathy, and abnormal results for neurography, temperature detection, or heart rate variability. Test-retest examination of TDD was performed in nine of the patients. RESULTS: Twenty-six of the patients had abnormal TDD (sensitivity 0.60) and 20 had abnormal vibration detection (sensitivity 0.46). Ten of the patients had abnormal TDD and normal vibration detection. Four of the patients had abnormal vibration detection and normal TDD. Test-retest examination of TDD showed a high degree of reproducibility (r = 0.87). CONCLUSION: TDD seems more useful than vibration detection in examination of diabetic neuropathy. PMID- 19804479 TI - Evaluation of multiple sclerosis diagnostic criteria in Suzhou, China--risk of under-diagnosis in a low prevalence area. AB - OBJECTIVE: To evaluate the discharge diagnosis of demyelinating diseases in the central nervous system (CNS) and analyze the predictive value of the new diagnostic criteria in Suzhou, China. MATERIALS AND METHODS: We collected clinical information and data of laboratory examinations for all cases with a diagnosis of various demyelinating diseases in the CNS. All data were reviewed individually by four senior neurologists, and a diagnosis was finally given to each patient according to the McDonald criteria and the Poser criteria for multiple sclerosis (MS). RESULTS: In the analysis, 176 patients with a diagnosis of demyelinating diseases in the CNS at discharge were included. In 82 patients with a diagnosis of MS at discharge, the MS diagnosis was confirmed for 74 patients according to the McDonald criteria for MS, and the positive predictive value for the discharge diagnosis of MS was 90.2% (74/82). According to the Poser criteria, 61 patients were diagnosed as MS. The consistency of the two diagnostic criteria for MS was 78.4%, based on the results of the evaluation. CONCLUSIONS: Under-diagnosis of MS could be one of the explanations for the low prevalence of MS in China. Compared to the Poser criteria, the McDonald criteria had a higher sensitivity for the diagnosis of MS. PMID- 19804480 TI - Disseminated Geotrichum candidum infection in a patient with relapsed acute myelogenous leukemia following allogeneic stem cell transplantation and review of the literature. AB - We describe a woman with relapsed acute myelogenous leukemia after allogeneic stem cell transplantation who developed disseminated Geotrichum candidum infection during chemotherapy-induced neutropenia. The isolate was susceptible to voriconazole, amphotericin B, and micafungin in vitro. We review the literature regarding invasive infections with G. candidum, which predominantly affect immunocompromised hosts, and discuss potential therapies for this rare pathogen. PMID- 19804481 TI - Blood transfusion as an alternative bartonellosis transmission in a pediatric liver transplant. PMID- 19804482 TI - Would patients with hepatitis C virus on the waiting list for a liver transplant accept a hepatitis C-positive organ? PMID- 19804483 TI - PAR4: a new role in the modulation of visceral nociception. AB - Protease-activated receptors (PARs) are a family of G-protein-coupled receptors with a widespread distribution that are involved in various physiological functions including inflammation and nociception. In a recent study in Neurogastroenterology and Motility, Auge et al. describe for the first time the presence of PAR4 on visceral primary afferent neurons and its role in modulating colonic nociceptive responses, colonic hypersensitivity and primary afferent responses to PAR2 and Transient Receptor Potential Vanilloid-4 (TRPV4). Using the model of visceromotor response (VMR) to colorectal distension (CRD), they show that a PAR4 agonist delivered into the colon lumen decreases basal visceral response to CRD and reduces the exacerbated VMR to CRD induced by treatment with PAR2 or TRPV4 agonists. In isolated sensory neurons, they show that a PAR4 agonist inhibits calcium mobilization induced by PAR2 or TRPV4 agonists. Finally, they describe increased pain behaviour evoked by luminal application of mustard oil in PAR4 deficient mice compared to wild type controls. The newly discovered role of PAR4 in modulating visceral pain adds to our growing understanding of the contribution of colonic proteases and PARs to the mechanisms involved in colonic hypersensitivity and their potential role as therapeutic targets for irritable bowel syndrome. PMID- 19804484 TI - Anopheles gambiae innate immunity. AB - The successful development of Plasmodium in Anopheles mosquitoes is governed by complex molecular and cellular interactions that we are just beginning to understand. Anopheles immune system has received particular attention as genetic evidence points clearly to its critical role in eliminating the majority of parasites invading the midgut epithelium. Several factors regulating Plasmodium development have been identified and tentatively assigned to the individual steps leading to mosquito immune reactions; non-self-recognition, signal modulation, signal transduction and effector mechanisms. Detailed knowledge of these steps and their underlying molecular mechanisms may offer novel perspectives to abort Plasmodium development in the vector. Here, we summarize our current knowledge of mosquito innate immunity highlighting both, recent advances and areas where additional research is required. PMID- 19804485 TI - Passing GO (gene ontology) in plant pathogen biology: a report from the Xanthomonas Genomics Conference. AB - In mid-July a workshop entitled the 'Xanthomonas Genomics Conference' took place at the stunning location of Pingree Park, Colorado State University, USA. This meeting, which was supported this time round by United States Department of Agriculture and US National Science Foundation, was the third official workshop dedicated to the study of phytopathogens belonging to the species Xanthomonas. One of the major goals of this meeting was to discuss the insight that comparative analysis of Xanthomonas genomes has given both to an understanding of the ability of this important group of bacteria to exploit an extraordinary diversity of plant hosts and host tissues, and to the development of needed improvements in disease control and prevention. In this report we give an overview of recent developments in this field that were presented during the meeting. These highlights included the unveiling of 11 new Xanthomonas genomic sequences, structural and functional insights into the peptide Ax21 elicitor, the first description of small non-coding RNAs in Xanthomonas and the role they play in the regulation of virulence, as well as a description of novel type III secreted effectors which target different hosts. PMID- 19804486 TI - Modulation of NF-kappaB activation in Theileria annulata-infected cloned cell lines is associated with detection of parasite-dependent IKK signalosomes and disruption of the actin cytoskeleton. AB - Apicomplexan parasites within the genus Theileria have the ability to induce continuous proliferation and prevent apoptosis of the infected bovine leukocyte. Protection against apoptosis involves constitutive activation of the bovine transcription factor NF-kappaB in a parasite-dependent manner. Activation of NF kappaB is thought to involve recruitment of IKK signalosomes at the surface of the macroschizont stage of the parasite, and it has been postulated that additional host proteins with adaptor or scaffolding function may be involved in signalosome formation. In this study two clonal cell lines were identified that show marked differences in the level of activated NF-kappaB. Further characterization of these lines demonstrated that elevated levels of activated NF kappaB correlated with increased resistance to cell death and detection of parasite-associated IKK signalosomes, supporting results of our previous studies. Evidence was also provided for the existence of host- and parasite-dependent NF kappaB activation pathways that are influenced by the architecture of the actin cytoskeleton. Despite this influence, it appears that the primary event required for formation of the parasite-dependent IKK signalosome is likely to be an interaction between a signalosome component and a parasite-encoded surface ligand. PMID- 19804487 TI - Right ventricular septal pacing: the success of stylet-driven active-fixation leads. AB - BACKGROUND: The detrimental effects of right ventricular (RV) apical pacing on left ventricular function has driven interest in alternative pacing sites and in particular the mid RV septum and RV outflow tract (RVOT). RV septal lead positioning can be successfully achieved with a specifically shaped stylet and confirmed by the left anterior oblique (LAO) fluoroscopic projection. Such a projection is neither always used nor available during pacemaker implantation. The aim of this study was to evaluate how effective is the stylet-driven technique in septal lead placement guided only by posterior-anterior (PA) fluoroscopic view. METHODS: One hundred consecutive patients with an indication for single- or dual-chamber pacing were enrolled. RV septal lead positioning was attempted in the PA projection only and confirmed by the LAO projection at the end of the procedure. RESULTS: The RV lead position was septal in 90% of the patients. This included mid RV in 56 and RVOT in 34 patients. There were no significant differences in the mean stimulation threshold, R-wave sensing, and lead impedance between the two sites.In the RVOT, 97% (34/35) of leads were placed on the septum, whereas in the mid RV the value was 89% (56/63). CONCLUSIONS: The study confirms that conventional active-fixation pacing leads can be successfully and safely deployed onto the RV septum using a purposely shaped stylet guided only by the PA fluoroscopic projection. PMID- 19804488 TI - Nontraditional implantable cardioverter defibrillator placement in adult patients with limited venous access: a case series. AB - BACKGROUND: Conventional transvenous approaches for implantable cardioverter defibrillator (ICD) lead placement are not possible in some patients with limited venous access or severe tricuspid valve dysfunction. METHODS: We retrospectively identified six patients who underwent ICD placement or revision requiring nontraditional alternative surgical lead placement at our institution between November 2006 and August 2008. The baseline and operative patient characteristic data were accumulated and reviewed. RESULTS: All the patients (mean age 71 +/- 3.4 years) underwent nontraditional surgical placement of epicardial ICD leads and traditional placement of ventricular epicardial bipolar pacing/sensing leads. Five patients had the distal lead tip fixed to the anterior epicardium of the right ventricular outflow tract, which was then looped under and around the ventricles, forming a "sling," and tunneled to a left subclavicular pocket. One patient had a single unipolar subcutaneous array lead fashioned into a "loop" and placed under the inferior aspect of the ventricles. The average procedure time was 311 +/- 115 minutes with a mean defibrillatory threshold (DFT) of < or = 22 + 3 J. Post-procedure hospitalization was 9.3 +/- 4.4 days and no device-related complications were encountered. Mean device follow-up of 451 + 330 days showed normal function and two appropriate successful ICD discharges. CONCLUSION: Nontraditional alternative surgical methods for the placement of ICD systems in adult patients with limited venous access or TV dysfunction can achieve results similar to those of conventionally placed endovascular leads with limited complications and comparable DFTs in short-term follow-up. PMID- 19804489 TI - Impact of temporary interruption of right ventricular pacing for heart block on left ventricular function and dyssynchrony. AB - BACKGROUND: The increasing data suggest an association between chronic right ventricular (RV) and left ventricular (LV) dysfunction. We sought to determine the effect of temporary interruption of long-term RV pacing on LV function and mechanical dyssynchrony in children and young adults with complete heart block. METHODS: Twelve patients aged 20.0 + or - 7.4 years with congenital heart block (group I) and six patients aged 22.7 + or - 11.0 years with surgically acquired heart block (group II) with RV pacing were studied. The pacing rate was reduced to less than patient's intrinsic heart rate and maintained for 5 minutes. The LV ejection fraction (EF), three-dimensional systolic dyssynchrony index (SDI), two dimensional global longitudinal strain and strain rate, and Doppler-derived isovolumic acceleration before and after interruption of RV pacing were compared. RESULTS: The LVEF and GLS increased while QRS duration decreased after the pacing interruption in both the groups (all P < 0.05). While SDI decreased in both groups I (6.8 + or - 2.3%- 3.8 + or - 0.8%, P = 0.001) and II (9.2 + or - 4.1% 5.0 + or - 1.6%, P = 0.032), it remained higher in group II than in group I (P = 0.046) after the pacing interruption. The prevalence of LV dyssynchrony (SDI > 4.7%) decreased in group I (83%-25%, P = 0.006) but not in group II (67%-50%, P = 0.50). The %increase in LVEF correlated positively with %reduction of LV SDI (r = 0.80, P = 0.001). CONCLUSIONS: Temporary interruption of chronic RV pacing acutely improves LV dyssynchrony and systolic function in children and young adults, the magnitude of which is greater in patients with congenital than those with surgically acquired heart block. PMID- 19804490 TI - Idiopathic premature ventricular contractions exhibiting preferential conduction within the aortic root. AB - A 65-year-old man with frequent premature ventricular contractions (PVCs) underwent electrophysiological testing. Although an excellent pace map was obtained from the right coronary cusp (RCC), radiofrequency ablation at that site interrupted the PVCs transiently. Successful ablation was achieved in the left coronary cusp with earlier local ventricular activation during the PVCs than that in the RCC. These findings suggest that preferential conduction within the aortic root may exist and cause ventricular arrhythmias (VAs) arising from this region to exhibit variable electrocardiographic features, thereby limiting the reliability of electrocardiographic algorithms and pace mapping to predict the site of the VA origin. PMID- 19804491 TI - Reflections on industry-supported-continuing-medical educational activities for cardiac electrophysiology trainees. PMID- 19804492 TI - Permanent pacemaker implantation following aortic valve replacement: current prevalence and clinical predictors. AB - BACKGROUND: The incidence of conduction disease requiring permanent pacemaker (PPM) implantation following aortic valve replacement (AVR) ranges from 3% to 6%. Data concerning the potential risks for PPM requirement associated with certain valve types have been conflicting and controversial. We sought to evaluate the prevalence, predictors for PPM implantation, and PPM dependency during follow-up in patients undergoing AVR. METHODS: A total of 214 consecutive patients undergoing AVR were studied retrospectively. A total of 207 patients were included in the statistical analysis. Clinical variables including valve size and types were catalogued and the incidence of PPM evaluated. Cardiac rhythm device clinic records were examined and PPM dependency status was catalogued. Multivariate analyses were performed to determine predictors of PPM implantation and PPM dependency during follow-up. RESULTS: Fifteen patients (7.2%) required PPM postoperatively. After controlling for clinical and surgical characteristics, predictors for PPM included preoperative first-degree atrioventricular block with and without left anterior fascicular block or intraventricular conduction delay [odd ratios (OR) = 12.5, P = 0.001], cardiac arrest postoperatively (OR = 9.4, P = 0.012), and combined aortic and mitral valve surgery (OR = 11.5, P = 0.027). Aortic valve types did not predict complete heart block (CHB) and PPM implantation. Of those patients who underwent PPM implantation, 70% were classified as PPM dependent during long-term follow-up. CONCLUSION: CHB and PPM implantation continue to be common complications of AVR. Preexisting atrioventricular with intrafascicular or intraventricular conduction disease along with cardiac arrest and dual valve surgery are the most important significant predictors of PPM implantation and PPM dependency during follow-up. The selection of valve types did not predict conduction disease requiring PPM implantation. PMID- 19804493 TI - Allopurinol-induced DRESS syndrome in an adolescent patient. AB - A 16-year-old male patient, with a history of essential hypertension enrolled in an experimental drug protocol using allopurinol, presented to our emergency department with a 10-day history of fever. Initial laboratory evaluation revealed leukocytosis, eosinophilia, and transaminitis. After extensive work-up and exclusion of infectious and oncologic etiologies, the diagnosis of allopurinol induced drug reaction and eosinophilia with systemic symptoms syndrome was carried out. The patient responded to administration of IV methylprednisolone, with complete resolution of symptoms and improvement of laboratory abnormalities. This case represents the first report of allopurinol-induced drug reaction and eosinophilia with systemic symptoms syndrome in a pediatric patient. PMID- 19804494 TI - Eccrine squamous metaplasia and periadnexal granulomas: new cutaneous histopathologic findings in cardiofaciocutaneous syndrome. AB - Cardiofaciocutaneous syndrome is a rare genetic disorder characterized by dysmorphic facial features and neurologic, cardiac, ophthalmologic, and dermatologic findings. Previously reported skin and hair findings in cardiofaciocutaneous syndrome include sparse, slow-growing curly hair, atopic dermatitis, ichthyosis, follicular hyperkeratosis, and keratosis pilaris. We report the case of a 4-year-old boy who has cardiofaciocutaneous syndrome with previously unreported histopathologic findings of eccrine squamous metaplasia and periadnexal granuloma. PMID- 19804495 TI - Steroid-induced periorificial dermatitis in children--clinical features and response to azelaic acid. AB - Periorificial dermatitis, a common skin disease in young women, has been occasionally reported in children. This study elaborates the clinical features of periorificial dermatitis in children as well as possible pathogenetic factors and the response to 20% azelaic acid cream. A total of 10 children aged 3 to 12 (mean 7.7) years suffering from nongranulomatous periorificial dermatitis for 3 to 7 (mean 4.9) months were evaluated, and dermatologic examination was carried out. Pretreatment was documented. Skin prick tests with a panel of six common inhalative allergens and patch tests with the European Standard Series were performed. An association between atopy and periorificial dermatitis was evaluated, and patients were screened for skin colonization by fungi, bacteria, and Demodex mites. They were treated with 20% azelaic acid cream, which was topically applied twice daily on all affected areas, until complete resolution was achieved. Treatment period was followed by an individual observation period. Periorificial dermatitis had developed in typical distribution and morphology. In all patients, low- to high-potency topical corticosteroids had been used on the face prior to manifestation. Atopy was found in half of the patients. Allergological, bacteriological, and mycological examinations did not reveal pathologic results. Demodex mites could not be demonstrated by skin surface biopsy. Treatment with 20% azelaic acid cream led to complete resolution of skin lesions after 4 to 8 (mean 5.4) weeks in all patients. Transient exacerbation of skin condition with a peak between the 2nd and 6th day of treatment could be observed in three patients. Side effects of 20% azelaic acid cream were registered in six patients and were predominantly present in the first 2 weeks of treatment. Side effects were minimal and became rarer with ongoing treatment. No recurrences were seen within a follow-up period of 2 to 8 (mean 4.4) months. Treatment with 20% azelaic acid cream could provide an effective and safe alternative therapeutic option in children with nongranulomatous periorificial dermatitis. PMID- 19804496 TI - Childhood vitiligo: a long-term study of localized vitiligo treated by noncultured cellular grafting. AB - Segmental type was the second most commonly reported in childhood vitiligo. No significant difference has been reported in the prevalence of childhood and adult focal vitiligo. However, the prevalence of segmental vitiligo has been found to be higher in children compared with that in adults. All available medical and phototherapy options are limited by adverse effects or unsatisfactory efficacy. Surgical techniques may be preferred but are not recommended for children as they are time consuming and associated with technical difficulties. In a retrospective review, 25 children aged 4 to 16 years were treated by autologous, noncultured cellular grafting performed under sedation supplemented with local anaesthesia and were followed up for a period of 9 to 54 months postgrafting. Repigmentation was graded as excellent with 95% to 100% pigmentation, good with 65% to 94%, fair with 25% to 64%, and poor with 0% to 24% of the treated area. In the segmental group, eight (62%) showed excellent, two (15%) good, one (8%) fair, and two (15%) poor pigmentation, which was retained until the end of the respective follow-up period. In the focal group, nine (75%) showed excellent, and one (8%) each showed good, fair, and poor pigmentation, which was retained until the end of the respective follow-up period. Noncultured cellular grafting may be considered to treat childhood localized vitiligo. PMID- 19804497 TI - Comparative study on the efficacy, safety, and acceptability of imiquimod 5% cream versus cryotherapy for molluscum contagiosum in children. AB - To compare the efficacy, safety and acceptability of imiquimod (IMQ) 5% cream with cryotherapy for the treatment of molluscum contagiosum (MC) in children. Prospective, randomized, comparative, observer blinded study. A total of 74 children, with MC were divided randomly to receive treatment with either IMQ 5% cream (group A) 5 days a week or cryotherapy (group B) once a week until clinical cure or up to a maximum of 16 weeks. All the patients were followed up weekly during active treatment. The patients were followed-up for 6 months after clinical cure to look for recurrence. In the IMQ group (group A), the overall complete cure rate was 91.8% (34 of 37), 22 of the 37 patients cleared by the end of 6 weeks and 12 more patients cleared by the end of 12 weeks, while the remaining three patients (8.1%) did not clear even after 16 weeks. Whereas, in the cryotherapy group, all 37 patients achieved complete cure, 26 of 37 (70.27%) patients cleared after 3 weeks, and the remaining 11 (29.72%) cleared by the end of 6 weeks. No statistically significant difference was found between the overall complete cure rate in both groups at the end of maximum treatment period (16 weeks). Pain, bullae formation, pigmentary changes, and superficial scarring were more significantly common in the cryotherapy group compared with the IMQ group. Imiqimod 5% cream seems to be slow acting but an effective agent for the treatment of MC in children. IMQ appears to be practically painless and more cosmetically accepted treatment when compared with cryotherapy, and may be the preferred treatment of MC in children especially with numerous small lesions. Cryotherapy has the advantage of being rapidly effective, and is less expensive than IMQ and may be the preferred treatment for large solitary or few lesions. PMID- 19804498 TI - Infant skin microstructure assessed in vivo differs from adult skin in organization and at the cellular level. AB - Functional differences between infant and adult skin may be attributed to putative differences in skin microstructure. The purpose of this study was to examine infant skin microstructure in vivo and to compare it with that of adult skin. The lower thigh area of 20 healthy mothers (ages 25-43) and their biological children (ages 3-24 months) was examined using in vivo noninvasive methods including fluorescence spectroscopy, video microscopy, and confocal laser scanning microscopy. Stratum corneum and supra-papillary epidermal thickness as well as cell size in the granular layer were assessed from the confocal images. Adhesive tapes were used to remove corneocytes from the outer-most layer of stratum corneum and their size was computed using image analysis. Surface features showed differences in glyph density and surface area. Infant stratum corneum was found to be 30% and infant epidermis 20% thinner than in adults. Infant corneocytes were found to be 20% and granular cells 10% smaller than adult corneocytes indicating a more rapid cell turnover in infants. This observation was confirmed by fluorescence spectroscopy. Dermal papillae density and size distribution also differed. Surprisingly, a distinct direct structural relationship between the stratum corneum morphology and the dermal papillae was observed exclusively in infant skin. A change in reflected signal intensity at approximately 100 mum indicating the transition between papillary and reticular dermis was evident only in adult skin. We demonstrate in vivo qualitative and quantitative differences in morphology between infant and adult skin. These differences in skin microstructure may help explain some of the reported functional differences. PMID- 19804499 TI - Enhanced drought-tolerance in the homoploid hybrid species Pinus densata: implication for its habitat divergence from two progenitors. AB - The homoploid hybrid species Pinus densata is restricted to alpine habitats that exceed the altitude range of its two parental species, Pinus tabulaeformis and Pinus yunnanensis. Alpine habitats usually generate cold-induced water stress in plants. To understand the ecological differentiation between these three species, we examined their physiological responses to drought stress. Potted seedlings of three species were subjected to low, mild, moderate and severe water stress in an automatic-controlled glasshouse. Fifteen indicators of fitness were measured for each species in each treatment, and most of these decreased as drought increased. Pinus densata exhibited higher fitness than both parental species in terms of total dry mass production (TDM) and long-term water use efficiency (WUE(L)) across all treatments; several other ecophysiological traits were also extreme but not across every treatment, and not always in the highest stress treatment. These results indicate that extreme characters that have become well fixed in P. densata, confer a faster seedling growth rate and more efficient water use, which in turn should confer increased drought tolerance. These traits of P. densata likely promoted its ecological separation from its parental species and facilitated its successful colonization and establishment in high-altitude habitats. PMID- 19804500 TI - Hepatitis C virus NS4 protein impairs the Th1 polarization of immature dendritic cells. AB - Dendritic cells (DCs) in chronic hepatitis C patients display impaired function, although the details remain unclear. To investigate the hepatitis C virus (HCV) protein that has the most impact on DC function, we compared five recombinant proteins and seven HCV protein genes in modulating DC phenotype and function. Immature DCs (iDCs) were established from healthy donor peripheral blood monocytes with granulocyte-macrophage colony stimulating factor (GM-CSF) and IL 4. Lipopolysaccharide was used to establish mature DCs (mDCs). Cells were then pulsed with HCV recombinant proteins or transfected with HCV plasmids and subsequently assayed for cell surface marker expression by flow cytometry. For cytokine and proliferative T-cell response analysis, DCs were cultured with autologous CD4 T cells and tuberculin purified protein derivative (PPD). Mean fluorescent intensity of CD86 was reduced in HCV protein-pulsed iDCs. Proliferative T-cell responses and Th1 cytokine concentrations were reduced with HCV nonstructural proteins (NS), particularly with HCV NS4. HCV nonstructural proteins, particularly NS4, change the iDC phenotype and reduce antigen-specific T-cell stimulatory function with Th1 cytokine reductions. PMID- 19804501 TI - Optimal combinations of ultrasound-based and serum markers of disease severity in patients with chronic hepatitis C. AB - Combinations of noninvasive markers may improve discrimination of chronic liver disease severity. The aims of this study were to compare four validated serum and ultrasound-based markers of hepatic disease severity head-to-head with liver biopsy and to assess optimal combinations with consideration of cost. A total of 67 patients with biopsy-proven chronic hepatitis C underwent all four techniques on the same visit [aspartate aminotransferase (AST) to platelet ratio index (APRI); Enhanced Liver Fibrosis (ELF) panel; transient elastography (TE) and ultrasound microbubble hepatic transit times (HTT)]. Markers were combined according to increasing financial cost and ordinal regression used to determine contributions. APRI, ELF, TE and HTT predicted cirrhosis with diagnostic accuracy of 86%, 91%, 90% and 83% respectively. ELF and TE were the most reliable tests with an intra-class correlation of 0.94 each. Either ELF or TE significantly enhanced the prediction of fibrosis stage when combined with APRI, but when combined together, did not improve the model further. Addition of third or fourth markers did not significantly improve prediction of fibrosis. Combination of APRI with either ELF or TE effectively predicts fibrosis stage, but combinations of three or more tests lead to redundancy of information and increased cost. PMID- 19804502 TI - Clinical features of biochemical cholestasis in patients with recurrent hepatitis C after living-donor liver transplantation. AB - Recurrent hepatitis C after liver transplantation (HepC-LT) progresses faster than hepatitis C in non-transplant settings. Cholestasis has been suggested to be one characteristic of HepC-LT related to the rapid progression. We investigated the clinical features of biochemical cholestasis, which we defined as high serum concentrations of alkaline phosphatase and gamma-glutamyl transpeptidase, in patients with recurrent hepatitis C after living-donor liver transplantation. Eighty patients were diagnosed with post-transplant recurrent hepatitis C after exclusion of other aetiologies of cholestasis by liver biopsy and imaging. The clinical features of biochemical cholestasis in the patients with HepC-LT, including histological changes, the efficacy of interferon therapy and helper T cell (Th) subsets in the peripheral blood, were analysed. Fifty-five of the 80 patients with HepC-LT (69%) had evidence of biochemical cholestasis. Progression of liver fibrosis to stage F3 or F4 was significantly accelerated in patients with biochemical cholestasis compared with patients without cholestasis. The biochemical cholestasis in patients with HepC-LT improved after interferon therapy in 22 of 39 patients (56%) who showed a virological response to the therapy, suggesting that hepatitis C virus (HCV) caused the biochemical cholestasis in these patients. Patients with biochemical cholestasis who had a biochemical response to interferon therapy showed an increased Th1 responses in peripheral blood. In conclusion, biochemical cholestasis is the characteristic feature of HepC-LT and is related to progression of liver fibrosis. An increased Th1 response is associated with cholestasis caused by HCV after liver transplantation. PMID- 19804503 TI - Adverse effects of Sudanese toombak vs. Swedish snuff on human oral cells. AB - BACKGROUND: The high incidence of oral cancer in Sudan has been associated with the use of toombak, the local type of smokeless tobacco. However, its specific effects on human oral cells are not known. We aimed to investigate the effects of toombak on primary normal human oral keratinocytes, fibroblasts, and a dysplastic oral keratinocytic cell line, and to compare them with the effects induced by Swedish snuff. METHOD: Aqueous extracts were prepared from moist toombak and Swedish snuff and added in serial dilutions on in vitro monolayer cultured cells. Cell viability, morphology and growth, DNA double-strand breaks (gammaH2AX staining), expression of phosphatidylserine (Annexin V staining), and cell cycle were assessed after various exposure time periods. RESULTS: Significant decrease in cell number, occurrence of DNA double-strain breaks, morphological and biochemical signs of programmed cell death were detected in all oral cell types exposed to clinically relevant dilutions of toombak extract, although to a lesser extent in normal oral fibroblasts and dysplastic keratinocytes. G2/M-block was also detected in normal oral keratinocytes and fibroblasts exposed to clinically relevant dilutions of toombak extract. Swedish snuff extract had less adverse effects on oral cells, mainly at non-clinically relevant dilutions. CONCLUSION: This study indicates a potential for toombak, higher than for Swedish snuff, to damage human oral epithelium. Dysplastic oral keratinocytes were less sensitive than their normal counterparts, suggesting that they might have acquired a partially resistant phenotype to toombak-induced cytotoxic effects while still being prone to DNA damage that could lead to further malignant progression. PMID- 19804504 TI - Oral pain perception and taste in burning mouth syndrome. AB - BACKGROUND: Aim of this study was to investigate the effect of idiopathic burning mouth syndrome (BMS) on both, the pain perception within trigeminal lingual nerve distribution and gustatory sensitivity using capsaicin threshold test, and regional taste tests, respectively. METHODS: Pain thresholds for capsaicin were assessed using capsaicin-impregnated filter-paper strips. The strips were placed midline on the tongue tip for whole mouth testing with the mouth closed, and on the left or right edge of the extended anterior tongue for lateralized testing. Measures of gustatory function were obtained by validated "taste strips" test kit and electrogustometry. The tests were applied to 13 patients with BMS. Results were compared with those from 28 healthy subjects. RESULTS: Patients with BMS exhibited a decreased gustatory and somatosensory perception compared with healthy controls. These changes were found for lateralized tests but not for the whole mouth test procedure. Duration of disorder showed an effect on the capsaicin threshold, with patients being less sensitive to capsaicin exhibiting an increased duration of disorder. CONCLUSION: Both pain-related and gustatory sensitivies of the tongue are found to be decreased in BMS. PMID- 19804505 TI - The role of bone marrow-derived mesenchymal stem cells in treating formocresol induced oral ulcers in dogs. AB - BACKGROUND: Mesenchymal stem cells (MSCs), a subpopulation of adult somatic stem cells, are an attractive stem cell source in regenerative medicine because of their multipotentiality. In this study, the effects of MSCs transplantation on oral ulcer healing were examined. METHODS: Mesenchymal stem cells were isolated from bone marrow aspirates of dogs by dish adherence and expanded in culture. Oral ulcers were induced by topical application of formocresol in the oral cavity of dogs. Either autologous MSCs or vehicle (saline) was injected around the ulcer. The healing process of the ulcer was monitored clinically and histopathologically. Gene expression of vascular endothelial growth factor (VEGF) was detected in MSCs by reverse transcription-polymerase chain reaction. Expression of VEGF and collagen genes was detected in biopsies from all ulcers. RESULTS: Mesenchymal stem cells expressed mRNA for VEGF MSCs transplantation significantly accelerated oral ulcer healing compared with controls. There was increased expression of both collagen and VEGF genes in MSCs-treated ulcers compared with controls. CONCLUSION: Mesenchymal stem cells transplantation may help accelerate oral ulcer healing, possibly through the induction of angiogenesis by VEGF together with increased intracellular matrix formation as detected by increased collagen gene expression. PMID- 19804506 TI - QRS duration in the selection of patients for cardiac resynchronization therapy. PMID- 19804507 TI - Fragmented QRS complexes on 12-lead ECG: a marker of cardiac sarcoidosis as detected by gadolinium cardiac magnetic resonance imaging. AB - BACKGROUND: Fragmented QRS complexes (fQRS) on a 12-lead ECG are a marker of myocardial scar in patients with coronary artery disease. Cardiac sarcoidosis is also associated with myocardial granuloma formation and scarring. We evaluated the significance of fQRS on a 12-lead ECG compared to Gadolinium-delayed enhancement images (GDE) in cardiac magnetic resonance imaging (CMR). METHOD AND RESULTS: The ECGs of patients (n = 17, mean age: 52 +/- 11 years, male: 53%) with established diagnosis of sarcoidosis who underwent a CMR for evaluation of cardiac involvement were studied. ECG abnormalities included bundle branch block, Q wave, and fQRS. fQRS, Q wave, and bundle branch block were present in 9 (53%), 1 (6%), and 4 (24%) patients, respectively. The sensitivity and specificity of fQRS for detecting abnormal GDE were 100% and 80%, respectively. Sensitivity and specificity of Q waves were 11% and 100%, respectively. CONCLUSIONS: fQRS on a 12 lead ECG in patients with suspected cardiac sarcoidosis are associated with cardiac involvement as detected by GDE on CMR. PMID- 19804508 TI - Influence of heavy cigarette smoking on heart rate variability and heart rate turbulence parameters. AB - BACKGROUND: Cigarette smoking increases the risk of cardiovascular events related with several mechanisms. The most suggested mechanism is increased activity of sympathetic nervous system. Heart rate variability (HRV) and heart rate turbulence (HRT) has been shown to be independent and powerful predictors of mortality in a specific group of cardiac patients. The goal of this study was to assess the effect of heavy cigarette smoking on cardiac autonomic function using HRV and HRT analyses. METHODS: Heavy cigarette smoking was defined as more than 20 cigarettes smoked per day. Heavy cigarette smokers, 69 subjects and nonsmokers 74 subjects (control group) were enrolled in this study. HRV and HRT analyses [turbulence onset (TO) and turbulence slope (TS)] were assessed from 24-hour Holter recordings. RESULTS: The values of TO were significantly higher in heavy cigarette smokers than control group (-1.150 +/- 4.007 vs -2.454 +/- 2.796, P = 0.025, respectively), but values of TS were not statistically different between two groups (10.352 +/- 7.670 vs 9.613 +/- 7.245, P = 0.555, respectively). Also, the number of patients who had abnormal TO was significantly higher in heavy cigarette smokers than control group (23 vs 10, P = 0.006). TO was correlated with the number of cigarettes smoked per day (r = 0.235, P = 0.004). While LF and LF/HF ratio were significantly higher, standard deviation of all NN intervals (SDNN), standard deviation of the 5-minute mean RR intervals (SDANN), root mean square of successive differences (RMSSD), and high-frequency (HF) values were significantly lower in heavy smokers. While, there was significant correlation between TO and SDNN, SDANN, RMSSD, LF, and high frequency (HF), only HF was correlated with TS. CONCLUSION: Heavy cigarette smoking has negative effect on autonomic function. HRT is an appropriate noninvasive method to evaluate the effect of cigarette on autonomic function. Simultaneous abnormal HRT and HRV values may explain increased cardiovascular event risk in heavy cigarette smokers. PMID- 19804509 TI - Impact of preceding ventricular premature beats on heart rate turbulence. AB - BACKGROUND: Heart rate turbulence (HRT) has recently been introduced as a noninvasive tool for studying autonomic dysfunction. It presented short time fluctuation of sinus cycle length following single ventricular premature contraction (VPC). However, HRT parameters may be influenced by different factors. This study aimed to evaluate the possible influence of VPC frequency on HRT. METHODS: 24-h Holter recording was performed in patients with VPCs initially detected by 12-lead electrocardiography (ECG) in the outpatient department. The numbers of VPCs in 2- and 5-minute durations preceding each VPC tachogram were calculated. The HRT parameters and the numbers of the VPCs preceding VPC tachograms were analyzed. RESULTS: There were 23,122 available VPC tachograms from 107 healthy subjects included in the study. The turbulence onset (TO) value increased and the turbulence slope (TS) value decreased as VPC's frequency increased. The TO values rapidly increased when the number of VPCs was >15 beats in the 2-minute and >35 beats in the 5-minute durations. There was also a prominent decrease in TS values when the VPCs reached 14 and 30 beats in the 2- and 5-minute durations, respectively. CONCLUSION: Physiologic baroreflex may be attenuated under intensive stimulation, which is evidenced by blunted HRT parameters by frequent VPCs. Physiologic response to VPC's frequency may be related to baroreflex fatigue and is demonstrated as a sigmoid curve. PMID- 19804510 TI - Assessment of microvolt T-wave alternans in high-risk patients with the congenital long-QT syndrome. AB - BACKGROUND: Microvolt T-wave alternans (MTWA) has been used for arrhythmogenic risk stratification in cardiac disease conditions associated with increased risk of sudden cardiac death. Macroscopic T-wave alternans has been observed in patients with congenital long-QT syndrome (LQTS). The role of MTWA testing in patients with LQTS has not been established. OBJECTIVE: To determine the diagnostic value of MTWA testing in high-risk patients with LQTS. METHODS AND RESULTS: We assessed MTWA in 10 consecutive LQTS index patients who survived cardiac arrest or had documented torsade de pointes tachycardia and 6 first degree family members with congenital LQTS which had been genotyped in 13 of 16 subjects (7 index patients, 6 family members). No LQTS-causing mutation was identified in 3 index patients with overt QT prolongation. MTWA was assessed during standardized bicycle exercise testing using the spectral method and yielded negative (n = 8) or indeterminate (n = 2) results in index patients, respectively. Similarly, all first-degree family members tested MTWA negative except for one indeterminate result. Two genotype positive family members could not be tested (two children-4 and 9 years of age). CONCLUSION: In patients with congenital LQTS, free from structural heart disease and with a history of life threatening cardiac arrhythmias, assessment of MTWA does not yield diagnostic value. Hence, determination of MTWA in lower risk LQTS patients without spontaneous arrhythmic events is likely not to be useful for arrhythmia risk stratification. PMID- 19804511 TI - Electrocardiographic indices of left ventricular hypertrophy and repolarization phase share the same genetic influences: a twin study. AB - BACKGROUND: Both left ventricular hypertrophy (LVH) and repolarization phase (RP) are known to be attributable to genetic influences, but less is known whether they share same genetic influences. The aim of this study was to investigate to what extent individual differences in electrocardiographic (ECG) LVH and RP are explained by genetic and environmental influences and whether these influences are shared between these two traits. METHODS: Resting ECG recordings were obtained from 186 monozygotic and 203 dizygotic female twin individuals, aged 63 to 76 years. Latent factors, called LVH and RP, were formed to condense the information obtained from LVH indices (Cornell voltage and Cornell product) and T wave amplitudes (leads V(5) and II), respectively. Multivariate quantitative genetic modeling was used both to decompose the phenotypic variances into additive genetic, common environmental, and unique environmental influences, and for the calculation of genetic and environmental correlations between LVH and RP. RESULTS: Additive genetic influences explained 16% of individual differences in LVH and 74% in RP. The remaining individual differences were explained by both common and unique environmental influences. The genetic correlation and unique environmental correlation between LVH and RP were -0.93 and -0.05, respectively. CONCLUSIONS: In older women without overt cardiac diseases, RP is under stronger genetic control than LVH. The majority of genetic influences are shared between LVH and RP whereas environmental influences are mainly specific to each. PMID- 19804512 TI - Influence of diabetes and/or myocardial infarction on prevalence of abnormal T wave alternans. AB - BACKGROUND: Subjects with microvolt-level T-wave alternans (TWA) in association with structural heart disease have an increased risk for sudden cardiac death. The presence of diabetes (DM) is associated with an increased risk of sudden death but there is limited data on the impact of DM and previous myocardial infarction (MI) on TWA prevalence. METHODS: We performed a case-control cross sectional study in 140 patients referred for routine exercise testing within a large multispecialty clinic. All patients with a history of DM and MI status within the past year were eligible: group 1 (no DM or MI), group 2 (DM only), group 3 (MI only), group 4 (DM and MI). Patients performed a symptom-limited Bruce protocol exercise test with assessment of TWA by the spectral method using commercially available equipment. We used published criteria for the blinded interpretation of TWA; all tests not unequivocally negative were considered abnormal. RESULTS: Age and gender were similar in all groups. The prevalence of abnormal TWA in groups 1-4 was 24%, 20%, 48%, and 62%, respectively (between group P = 0.002). Logistic regression analysis in all patients showed that abnormal TWA was related to prior MI [OR (95% CI): 4.0 (1.8-8.9), P < 0.001] but not to prevalent DM [0.9 (0.4-1.8), P = 0.72]. In patients with DM, the prevalence of abnormal TWA was related to reduced ejection fraction (P = 0.034) but not to BMI, DM duration, glycemic control, insulin use, or the presence of microvascular complications. CONCLUSION: The presence of DM alone does not increase risk of abnormal TWA. Prospective studies are required to establish the prognostic value of TWA in patients with DM. PMID- 19804513 TI - Comparison between contrast-enhanced magnetic resonance imaging and Selvester QRS scoring system in estimating changes in infarct size between the acute and chronic phases of myocardial infarction. AB - BACKGROUND: The Selvester QRS score was developed as a method to estimate infarct size (IS) using the ECG and has been validated during the prereperfusion era. Few comparisons exist with contrast-enhanced magnetic resonance imaging (ceMRI) in reperfused patients. This study evaluates the ability of the Selvester QRS score to estimate serial changes in IS during the acute and chronic phases of the infarct evolution in patients who have received reperfusion therapy. METHODS: Thirteen patients with acute myocardial infarction underwent serial ceMRI studies in the acute (<1 week) and chronic phase (>2 months) after their initial myocardial infarction. QRS scoring was performed on the corresponding ECGs. The correlation between ceMRI measurement and QRS score estimation of IS was determined at both time points and for the difference between the two phases. RESULTS: The mean IS was 20.1 +/- 11.0% of total left ventricular mass (% LV) in the acute phase and 13.3 +/- 6.4% LV in the chronic phase ceMRI. The mean IS estimated by Selvester QRS score in the acute and chronic phases were 18.7 +/- 8.2% and 16.4 +/- 8.5% LV, respectively. A modest correlation was found for the acute (r = 0.57) and chronic phase IS (r = 0.54). However, there was no correlation for the difference in IS between the acute and chronic phases. CONCLUSIONS: In this pilot study, the Selvester QRS score correlates modestly to IS by ceMRI during both the acute and chronic phases of the infarction process. The serial changes over time in the Selvester QRS score and IS by ceMRI show no correlation. PMID- 19804514 TI - Assessment of physiological amplitude, duration, and magnitude of ECG T-wave alternans. AB - BACKGROUND: An association between T-wave alternans (TWA) and malignant ventricular arrhythmias is generally recognized. Because relatively low levels of TWA have also been observed in healthy (H) subjects, the question arises as to whether these are ascribable to noise and artifacts, or can be given the relevance of a physiological phenomenon characterizing a preclinical condition. METHODS: To answer this question, in the present study 20-minute not noisy, sinus ECG recordings, from 138 H-subjects and 148 coronary artery diseased (CAD) patients, were submitted to our adaptive match filter (AMF) procedure to identify and parameterize TWA in terms of duration (TWAD), amplitude (TWAA), and magnitude (TWAM, defined as the product of TWAD times TWAA). The 99.5th percentiles of mean values of TWAA, TWAD, and TWAM over 20-minute ECGs were used to define three threshold levels (THRD, THRA, and THRM), which allow discrimination of abnormal TWA levels. RESULTS: Nonstationary TWA was found in all our H-subjects and CAD patients. TWAD, TWAA, and TWAM levels were classified as being physiological in 99% of H-subjects and 87% of CAD-patients. A linear correlation (r =-0.52, P < 0.001) was found between TWAA and RR interval in the H-population. CONCLUSIONS: Our results support the hypothesis of the existence of physiological TWA levels, which are to be considered in the effort to improve reliability of nonphysiological TWA levels discrimination. PMID- 19804515 TI - Electrophysiologic characteristics of wide QRS complexes during pharmacologic termination of sustained supraventricular tachycardias with verapamil and adenosine: observations from electrophysiologic study. AB - BACKGROUND: In this study we evaluate wide QRS complexes observed during pharmacologic termination of supraventricular tachycardias. METHODS: Patients with supraventricular tachycardia, undergoing electrophysiologic study were enrolled. 12 mg of adenosine or 10 mg of verapamil were administered during tachycardia, under continuous monitoring of intaracardiac and surface electrocardiograms. Electrocardiographic features of ventricular ectopy were noted. RESULTS: Seventy-four patients were enrolled. 48 patients were randomized to adenosine and 26 to verapamil. Five different appearance patterns of ventricular ectopy were observed during termination of tachycardias. All wide QRS complexes were of ventricular origin and all of them were observed during the termination of tachycardia. Adenosine more frequently resulted in appearance of ventricular beats (15.4% vs 41.7%, P = 0.003), and this was more frequently observed in patients with atrioventricular nodal reentrant tachycardia. Patients with ventricular beats were younger than those without, in both, verapamil (47.5 +/- 15.6 vs 65.0 +/- 8.8 years, P = 0.04) and adenosine (40.9 +/- 13.8 vs 49.7 +/ 16.8, P = 0.03) groups. Left bundle branch block (LBBB)/superior axis morphology was most frequent morphology in adenosine group (55%). Two of 4 patients in verapamil group displayed LBBB/inferior axis QRS morphology and another 2 patients displayed LBBB/superior axis morphology. CONCLUSIONS: Noncatheter induced, five different appearance patterns and four distinct morphologies of ventricular origin were observed. Most of them do not directly terminate tachycardia, but are associated with its termination and are not observed in ongoing tachycardia. PMID- 19804516 TI - Correlation relationship assessment between left ventricular hypertrophy voltage criteria and body mass index in 41,806 Swiss conscripts. AB - INTRODUCTION: Electrocardiographic criteria for left ventricular hypertrophy (LVH) have been limited by low sensitivity at acceptable levels of specificity. A number of studies have demonstrated that body mass index (BMI) is associated with decreased sensitivity of ECG LVH classification in hypertensive patients. The objective of this study is to investigate the correlation relationship between LVH voltage criteria and BMI in Swiss conscripts. METHODS: A database of 41,806 young Swiss people, who underwent compulsory conscription for the Swiss Army, was compiled. Along with other medical data, an ECG was taken. Statistical analyses, such as linear regression and calculation of correlation coefficient, were carried out between LVH voltage criteria and BMI. RESULTS: The mean age in the studied population was 19.2 +/- 1.1 years with a median age of 19 years (range from 17 to 38 years). We found an overweight prevalence of 25.1%. The results showed that body habitus had significant association with Sokolow-Lyon voltages. A mean decrease of 13%, 5%, 19%, 14%, and 12% for the five studied Sokolow-Lyon indexes were found between normal range subjects (18.5 < or = BMI < 25) and obese subjects (25< or = BMI). CONCLUSIONS: Our study confirms the hypothesis that people with higher BMI, a growing section of the population, have lower ECG amplitudes. Therefore, the Sokolow-Lyon voltage criteria may underestimate the presence of LVH for subjects with higher BMI, which is not the case for the Cornell voltage. Our analysis suggests that computerized electrocardiography for the diagnosis of left ventricular hypertrophy based on Sokolow-Lyon voltages should incorporate the BMI factor. PMID- 19804517 TI - Technical mistakes during the acquisition of the electrocardiogram. AB - In addition to knowledge of normal and pathological patterns, the correct interpretation of electrocardiographic (ECG) recordings requires the use of acquisition procedures according to approved standards. Most manuals on standard electrocardiography devote little attention to inadequate ECG recordings. In this article, we present the most frequent ECG patterns resulting from errors in limb and precordial lead placement, artifacts in 12-lead ECG as well as inadequate filter application; we also review alternative systems to the standard ECG, which may help minimize errors. PMID- 19804518 TI - Termination of recurrent ventricular tachycardia by continuous positive airway pressure ventilation. AB - We report a case of hypertrophic cardiomyopathy with recurrent ventricular tachycardia that resolved after initiating continuous positive airway pressure therapy. PMID- 19804520 TI - Evidentiary standards for forensic anthropology. AB - As issues of professional standards and error rates continue to be addressed in the courts, forensic anthropologists should be proactive by developing and adhering to professional standards of best practice. There has been recent increased awareness and interest in critically assessing some of the techniques used by forensic anthropologists, but issues such as validation, error rates, and professional standards have seldom been addressed. Here we explore the legal impetus for this trend and identify areas where we can improve regarding these issues. We also discuss the recent formation of a Scientific Working Group for Forensic Anthropology (SWGANTH), which was created with the purposes of encouraging discourse among anthropologists and developing and disseminating consensus guidelines for the practice of forensic anthropology. We believe it is possible and advisable for anthropologists to seek and espouse research and methodological techniques that meet higher standards to ensure quality and consistency in our field. PMID- 19804521 TI - Fimag: the United Kingdom disaster victim/forensic identification imaging system. AB - Imaging is an integral diagnostic tool in mass fatality investigations undertaken traditionally by plain X-rays, fluoroscopy, and dental radiography. However, little attention has been given to appropriate image reporting, secure data transfer and storage particularly in relation to the need to meet stringent judicial requirements. Notwithstanding these limitations, it is the risk associated with the safe handling and investigation of contaminated fatalities which is providing new challenges for mass fatality radiological imaging. Mobile multi-slice computed tomography is an alternative to these traditional modalities as it provides a greater diagnostic yield and an opportunity to address the requirements of the criminal justice system. We present a new national disaster victim/forensic identification imaging system--Fimag--which is applicable for both contaminated and non-contaminated mass fatality imaging and addresses the issues of judicial reporting. We suggest this system opens a new era in radiological diagnostics for mass fatalities. PMID- 19804522 TI - The teaching of denture marking methods in dental schools in the United Kingdom and the United States. AB - Forensic organizations worldwide have recommended that dental prostheses should be marked with, at a minimum, the patient's name and preferably with further unique identifiers such as a social security number. The current study aimed to assess the denture marking practice of dental schools within the United States and the United Kingdom. A questionnaire-based survey was employed to gain both quantitative and qualitative data on the methods, practices, and ethos behind denture marking in 14 U.K. and 32 U.S. dental schools. One hundred percent of U.K. and 87.5% of U.S. schools returned surveys and the results suggest that, for dental schools where there is no legal or legislative need for denture marking, the practice is inconsistently taught and appears to be reliant on internal forces within the school to increase awareness. Among those schools practicing marking, only 18% employ a technique likely to withstand common postmortem assaults; this is a concern. PMID- 19804523 TI - The use of Hemastix and the subsequent lack of DNA recovery using the Promega DNA IQ system. AB - Following implementation of our automated process incorporating the Promega DNA IQ system as a DNA extraction method, a large number of blood-containing exhibits failed to produce DNA. These exhibits had been tested with the Hemastix reagent strip, commonly used by police investigators and forensic laboratories as a screening test for blood. Some exhibits were even tainted green following transfer of the presumptive test reagents onto the samples. A series of experiments were carried out to examine the effect of the Hemastix chemistries on the DNA IQ system. Our results indicate that one or more chemicals imbedded in the Hemastix reagent strip severely reduce the ability to recover DNA from any suspected stain using the DNA IQ magnetic bead technology. The 3,3',5,5' tetramethylbenzidine (TMB) used as the reporting dye appears to interact with the magnetic beads to prevent DNA recovery. Hydrogen peroxide does not seem to be involved. The Hemastix chemistries do not interfere in any way with DNA extraction performed using phenol-chloroform. The incompatibility of the Hemastix chemistries on the DNA IQ system forced us to adopt an indirect approach using filter paper to carry out the presumptive test. PMID- 19804524 TI - Santeria and Palo Mayombe: skulls, mercury, and artifacts. AB - Santeria and Palo Mayombe are syncretic religions created in the New World based upon African religious beliefs combined with Christianity. The main worship of Palo Mayombe involves religious receptacles that may contain earth, sticks, varied artifacts, and animal and human remains. Due to the use of human and nonhuman remains, discovery of these items often leads to involvement by the police due to a concern of homicide. We review in detail the medical examiner records of two of these ritualistic cases including the autopsy, anthropology, police, and investigators' reports. For the human remains, careful consideration of the context in which the remains were recovered, their state of preservation, and the associated artifacts (e.g., beads and mercury) are important in determining the appropriate level of forensic significance. Anthropological examination with particular attention to taphonomic characteristics also may help determine the origin and forensic significance. PMID- 19804525 TI - The stable isotope ratios of marijuana. II. Strontium isotopes relate to geographic origin. AB - Effectively addressing marijuana trade is aided by understanding marijuana geographic sources. We analyzed the (87)Sr/(86)Sr of marijuana samples grown in 79 counties across the United States to determine if a primary geologic signal is retained in marijuana, which could therefore be useful for geographic sourcing. The marijuana results were compared with modeled bedrock (87)Sr/(86)Sr values based on (87)Rb decay rates and a generalized geologic map of the U.S.A. A significant correlation was observed between marijuana (87)Sr/(86)Sr and modeled bedrock (87)Sr/(86)Sr. Although values clustered near the 1:1 relationship, there was a predominance of positive anomalies, perhaps attributable to carbonate bedrock. A small number of negative anomalies were also observed, which were generally associated with granitic bedrocks. These results suggest that strontium isotopes in marijuana record the geographic origins of marijuana, and that refinement of the base strontium map (or strontium isoscape) and improved understanding of other strontium sources would be productive. PMID- 19804526 TI - Forensic terrestrial photogrammetry from a single image. AB - Forensic terrestrial photogrammetry is one of the most valuable and low-cost resources of spatial data available today. Due to the ephemeral crime scene characteristics, these photographs can often capture information that is never to be seen again. This paper presents a novelty approach for the documentation, analysis, and visualization of crime scenes for which only a single perspective image is available. The photogrammetric process consists of a few well-known steps in close-range photogrammetry: features extraction, vanishing points computation, camera self-calibration, 3D metric reconstruction, dimensional analysis, and interactive visualization. Likewise, the method incorporates a quality control of the different steps accomplished sequentially. As a result, several cases of study are presented in the experimental results section in order to test their viability. The full approach can be applied easily through the free software, sv3DVision, which has been evaluated by a number of police officers, forensic scientists, and forensic educators satisfactorily. PMID- 19804527 TI - Skull fracture with brain expulsion in a one-level jumping-fall. AB - Here presented is the case of a one-level jumping-fall with extensive skull fractures and brain expulsion. The body was found on the basement floor at the foot of the stairs. At the autopsy, the skull was extensively fractured, with about half of the brain expulsed several feet away from the body. The cause of death was established as a craniocerebral trauma with brain expulsion. The circumstances and manner of death were still unclear at that time. A low fall seemed very unlikely considering the severity of the skull and brain damage. The police investigation clearly revealed that the man, in a paranoid psychotic state, attacked his wife with a knife and then was witnessed by his children to have hit his head several times with a hammer. Afterwards, they saw him running to the top of the basement stairs and jumping to the bottom of the stairs head first. PMID- 19804528 TI - An amino acid model for latent fingerprints on porous surfaces. AB - Analytical standards are needed in latent fingerprint detection for research and development as well as for quality control in routine work because normal fingerprints are too varied for comparison studies and tests. One way is to create latent fingerprints. For the amino acid sensitive detection method this can be achieved by coating test items with an amino acid solution using a modified commercial office bubble jet printer. Besides low costs, fast and easy preparation, the main advantage of a bubble jet printer is that the amino acid loading per area on the test item can be calculated by weighing the cartridge on a balance. This opens the possibility to determine the deviation for every printing series. The reproducibility of prints in a printing series made by one cartridge has a deviation of 2-16% and of prints made by different cartridges 20 25%. PMID- 19804529 TI - Medicolegal aspects of post-traumatic gastroduodenal ulcers: a retrospective study. AB - Retrospective examination of 5-year autopsy material showed the presence of posttraumatic gastroduodenal ulcers (PGDU) in 17.7% of decedents deemed to be at risk. They were more common in males (77%) and in patients aged over 50. In the majority of cases (76%) the survival period was < 12 days; in 16.5% it was < 48 h. PGDU developed most commonly in victims of polytrauma and isolated craniocerebral injury, with ISS values > or = 16; patients with spinal cord injuries were at greatest risk. Most frequently affected was the stomach, exhibiting numerous, usually superficial lesions, while solitary acute and exacerbated chronic peptic ulcers were more common in the duodenum. Complications of PGDU developed in 40% of cases, mostly in the form of hemorrhage; in 20% of cases PGDU have contributed to death. Medicolegal aspects of PGDU are, most frequently, concerned with the causal relationship between trauma, PGDU, and fatal outcome, as well as the potential for allegations of medical negligence. PMID- 19804530 TI - A LDR-PCR approach for multiplex polymorphisms genotyping of severely degraded DNA with fragment sizes <100 bp. AB - Reducing amplicon sizes has become a major strategy for analyzing degraded DNA typical of forensic samples. However, amplicon sizes in current mini-short tandem repeat-polymerase chain reaction (PCR) and mini-sequencing assays are still not suitable for analysis of severely degraded DNA. In this study, we present a multiplex typing method that couples ligase detection reaction with PCR that can be used to identify single nucleotide polymorphisms and small-scale insertion/deletions in a sample of severely fragmented DNA. This method adopts thermostable ligation for allele discrimination and subsequent PCR for signal enhancement. In this study, four polymorphic loci were used to assess the ability of this technique to discriminate alleles in an artificially degraded sample of DNA with fragment sizes <100 bp. Our results showed clear allelic discrimination of single or multiple loci, suggesting that this method might aid in the analysis of extremely degraded samples in which allelic drop out of larger fragments is observed. PMID- 19804531 TI - Subway train-related fatalities in New York City: accident versus suicide. AB - We examined the characteristics of subway train-related fatalities in New York City between Jan. 1, 2003 and May 31, 2007 in order to determine which factors are useful in differentiating accident from suicide. Subway train-related deaths with homicide and undetermined manners also are included. During this period, there were 211 subway train-related fatalities. The manners of death were: suicide (n = 111), accident (n = 76), undetermined (n = 20), and homicide (n = 4). The causes of death were blunt trauma (n = 206) and electrocution (n = 5). Torso transection and extremity amputation were more frequent in suicides. Antidepressant medications were more frequently detected in suicides, whereas cocaine and ethanol were more frequent in accidents. However, autopsy findings should be weighed in the context of the entire evaluation along with other circumstantial and investigative findings. In unwitnessed deaths where additional information is unavailable or discrepant, the most appropriate manner of death usually is undetermined. PMID- 19804532 TI - Study of short- and long-term storage of teeth and its influence on DNA. AB - DNA degradation can interfere with the resolution of forensic cases. Allelic dropout often reduces the opportunity for adequate comparisons between degraded and reference samples. This study analyzed DNA degradation in 24 extracted teeth after storage at room temperature for 0, 2, 5, and 10 years. DNA concentration, quantified by dot-blot hybridization, declined significantly for the first 2 years, but there was no significant further degradation from the second to the tenth year of storage. COfiler analysis was used and the allelic dropout ratio for the amelogenin locus relative to CSF1PO locus was also estimated. Statistically significant differences were found between fresh teeth and teeth from the 2- and 5-year groups but not from the 10-year group. Under our storage conditions most of the DNA degradation occurred during the first 2 years. Further research is needed to control for individual and external factors that could affect DNA. PMID- 19804533 TI - Pharmacokinetics and safety of fibrinogen concentrate. AB - BACKGROUND: Although fibrinogen concentrate has been available for the treatment of congenital fibrinogen deficiency for years, knowledge of its pharmacokinetics comes from only two small studies. OBJECTIVES: To assess the pharmacokinetic (PK) profile, clot integrity and safety of fibrinogen concentrate (human) (FCH) in patients with afibrinogenemia. PATIENTS AND METHODS: A multinational, prospective, open-label, uncontrolled study of patients with afibrinogenemia > or = 6 years of age was conducted in the USA and Italy. Plasma was collected before and after infusion for PK analyses and evaluation by rotational thromboelastometry of maximum clot firmness (MCF) to assess clot integrity. Safety was assessed on the basis of adverse events and laboratory parameters. RESULTS: After a single dose of 70 mg kg(-1) body weight (b.w.) FCH in 14 patients, median incremental in vivo recovery was a 1.7 mg dL(-1) increase per mg kg(-1) b.w., and median levels were 1.3 g L(-1) for fibrinogen activity and antigen 1 h after infusion. Median half-life (t(1/2)) was 77.1 h for fibrinogen activity and 88.0 h for antigen. Plasma recovery in children < 16 years old was similar to that in adults aged 16 to < 65 years, but the t(1/2) and area under the curve were decreased, with an increased steady-state volume and clearance. MCF increased by a mean of 8.9 mm from baseline to 1 h after infusion of FCH (P < 0.0001). All four adverse events reported were mild, and none was serious or related to study drug. CONCLUSIONS: These PK findings confirm a rapid increase in plasma fibrinogen levels after infusion with FCH. Together with the clot integrity and safety data and published data on efficacy, the results support the idea that FCH substitution can restore hemostasis with a good safety profile. PMID- 19804534 TI - Clinical guidelines and off-license recombinant activated factor VII: content, use, and association with patient outcomes. AB - BACKGROUND: Recombinant activated factor VII (rFVIIa) is increasingly being used off-license for treating critical bleeding. Guidelines may therefore be useful for improving processes and outcomes. Little is known regarding guidelines for off-license rFVIIa or their association with patient outcomes. OBJECTIVES: To investigate the availability of hospital guidelines for off-license rFVIIa use and the association between these guidelines and mortality. METHODS: Data were extracted from the Haemostasis Registry, which collects all cases of off-license rFVIIa use in participating institutions in Australia and New Zealand. Contributing hospitals were requested to supply a copy of the institutional guideline relating to off-license rFVIIa administration. The characteristics of patients treated in accordance with all elements of the guidelines were compared with those of patients for who one or more guideline elements had been violated. The relationship between guideline-directed treatment and 28-day mortality was investigated using stepwise logistic regression. RESULTS: Two thousand five hundred and fifty-one patients in 75 hospitals were available for analysis. Of these hospitals, 58 provided a guideline for analysis. Patients complying with all guideline elements (n = 530) did not differ from patients receiving care that violated guidelines (n = 1035) regarding age, size of dose, or gender. Guideline directed treatment was not found to have an association with 28-day mortality following logistic regression. CONCLUSIONS: Few patients are treated in accordance with the criteria of rFVIIa guidelines, despite their availability in the majority of hospitals. Moreover, 28-day mortality does not appear to be associated with the use of guidelines in this patient group. Refinement of guidelines relating to the off-license use of rFVIIa is therefore required. PMID- 19804535 TI - Effects of losartan, HO-1 inducers or HO-1 inhibitors on erectile signaling in diabetic rats. AB - INTRODUCTION: Activation of the renin-angiotensin system which is common in diabetes mellitus might affect heme oxygenase (HO-1) gene expression. AIM: Assessment of the effects of administration of angiotensin II (Ang II) receptor antagonist (losartan) with HO-1 inducer or inhibitor on erectile signaling in diabetic rats. MATERIALS AND METHODS: Seventy male rats were divided equally into seven groups; healthy controls, streptozotocin-induced diabetic rats, rats on citrate buffer, diabetic rats on losartan, diabetic rats on HO-1 inducer (cobalt protoporphyrin [CoPP]), diabetic rats on losartan and CoPP, and diabetic rats on losartan and HO-1 inhibitor (stannus mesoporphyrin [SnMP]). MAIN OUTCOME MEASURE: HO enzyme activity, HO-1 gene expression, cyclic guanosine monophosphate (cGMP) assay, intracavernosal pressure (ICP), and cavernous tissue sinusoids surface area. RESULTS: HO-1 gene expression, HO enzymatic activity, and cGMP were significantly decreased in the cavernous tissue of diabetic rats. These parameters were significantly elevated with the use of CoPP that restored the normal control levels of HO enzyme activity. Administration of losartan exhibited a significant enhancing effect on these parameters compared with the diabetic group, but not restored to the control levels, whereas administration of CoPP combined with losartan led to the restoration of their normal levels. ICP demonstrated significant decline in diabetic rats. The use of CoPP and/or losartan led to its significant improvement compared with diabetic rats. Administration of either losartan and/or CoPP led to a significant increase in the cavernous sinusoids surface area of diabetic rats. Administration of losartan with SnMP significantly decreased the enhancing effect of losartan on the studied parameters. CONCLUSION: The decline in erectile function in diabetes mellitus could be attributed to the downregulation of HO-1 gene expression. HO-1 induction added to Ang II receptor antagonist could improve erectile function. PMID- 19804536 TI - Transcriptional analysis of virulence-related genes in enterococci from distinct origins. AB - AIMS: The role of enterococci in food fermentation and as probiotics counteracts with their increasing importance as human pathogens. Over the years, several virulence factors have been described, mainly in clinical strains but also in food isolates. However, differential expression of such traits may modulate the pathogenic potential of the harbouring enterococci. To further unravel such differential response, this study aims to identify environmental cues responsible for triggering the expression of virulence-related genes. METHODS AND RESULTS: The differential expression of eight virulence-related genes (cylMBAI, agg, esp, efaA(fs) and efaAfm) in 16 enterococci from distinct origins, grown in conditions simulating environmental colonization and infection sites, was analysed by reverse transcriptase PCR. The expression profiles were environmental and strain dependent, because no constant response was observed neither for clinical nor food enterococci. CONCLUSIONS: Virulence expression profiles are strain-specific and unrelated with strain's origin or species allocation. SIGNIFICANCE AND IMPACT OF THE STUDY: The current study constitutes the first approach aimed at the evaluation of the differential expression of enterococcal virulence-related genes combining so many growth environments, enterococcal species and origins. So, with this investigation, we intend to contribute to the clarification of enterococcal pathogenicity potential, especially for food strains. PMID- 19804537 TI - Survival and gene expression of enterotoxigenic Escherichia coli during long-term incubation in sea water and freshwater. AB - AIMS: In this study, the main objective was to verify the hypothesis of induction of 'viable but non-culturable' (VBNC) forms of enterotoxigenic Escherichia coli (ETEC) during incubation in water. METHODS AND RESULTS: Six clinically isolated ETEC strains were studied. Viable counts showed culturable ETEC bacteria for up to 3 months in freshwater but only two out of six strains were culturable in seawater at this time point. Although the bacterial cells remained intact, no production or secretion of heat-labile (LT) or heat-stable (ST) enterotoxins was observed using GM1-ELISA methods. However, genes encoding ETEC toxins (STh and LT), colonization factors (CS7 and CS17), gapA and 16S RNA were expressed during 3 months in both sea water and freshwater microcosms as determined by real-time RT-PCR on cDNA derived from the bacteria. CONCLUSIONS: Clinically isolated ETEC strains can survive for long periods in both sea water and freshwater. The bacterial cells remain intact, and the gene expression of virulence genes and genes involved in metabolic pathways are detected after 3 months. SIGNIFICANCE AND IMPACT OF THE STUDY: These results indicate that ETEC bacteria can enter a VBNC state during stressful conditions and suggest that ETEC has the potential to be infectious after long-term incubation in water. PMID- 19804538 TI - Utilization of oligo- and polysaccharides at microgram-per-litre levels in freshwater by Flavobacterium johnsoniae. AB - AIMS: To obtain a bacterial strain that can be used to quantify biodegradable polysaccharides at concentrations of a few micrograms per litre in freshwater. METHODS AND RESULTS: Flavobacterium johnsoniae strain A3 was isolated from tap water supplemented with laminarin, pectin or amylopectin at 100 microg C l(-1) and river Rhine water. The organism utilized 14 of 23 oligo- and polysaccharides, and 1 of 9 monosaccharides, but none of the sugar acids, sugar alcohols, carboxylic acids or aromatic acids tested at 10 microg C l(-1). Amino acids promoted growth of strain A3, but not in coculture with assimilable organic carbon (AOC) test strain Pseudomonas fluorescens P17, which utilized these compounds more rapidly than strain A3. Compounds released by strain P17 and AOC test strain Spirillum sp. NOX grown on acetate promoted the growth of strain A3 at N(max) values of > or = 2 x 10(5) CFU ml(-1) of strain P17 and > or = 5 x 10(5) CFU ml(-1) of strain NOX. Significant growth of strain A3 was observed in surface water and in tap water in the presence of strain P17 (N(max) P17 < 2 x 10(5) CFU ml(-1)). CONCLUSIONS: Strain A3 utilizes oligo- and polysaccharides at microgram-per-litre levels. In surface water and in tap water, the organism was able to utilize compounds that were not utilized by strain P17. These compounds may include oligo- and/or polysaccharides. SIGNIFICANCE AND IMPACT OF THE STUDY: Phytoplanktonic and bacterial polysaccharides can constitute an important biodegradable fraction of natural organic matter in water and may promote growth of heterotrophic bacteria during water treatment and drinking water distribution. Strain A3 can be used to quantify a group of compounds that includes oligo- and polysaccharides at microgram-per-litre levels in freshwater. PMID- 19804539 TI - Effects of competition on great and blue tit reproduction: intensity and importance in relation to habitat quality. AB - 1. In studies on the effect of competition in plant communities two terms are used to describe its effects: the absolute reduction in growth of an individual as a consequence of the presence of another one is called intensity, while the relative impact of competition on an individual as a proportion of the impact of the whole environment is called importance. One school of thought is that the role of competition remains constant across productivity gradients, while the other is that it decreases with increasing severity. J.B. Grace (1991. A clarification of the debate between grime and tilman. Functional Ecology, 5, 583 587.) suggested that the apparent contradiction might be solved if we acknowledge that the two schools are discussing different aspects of competition: the intensity of competition might remain constant while its importance declines with increasing severity. 2. There are no studies that compare intensity and importance of competition in bird populations between areas that differ in quality or productivity and hence it is not possible to make predictions how intensity or importance of competition would vary between them. 3. I compared variation in intensity and importance of competition of three demographic variables between five plots that differ strongly in quality for great Parus major L. and blue tit Cyanistes caeruleus (L.). 4. Both intensity and importance of competition are larger in great than in blue tit populations meaning that the effect of competition on demographic variables is stronger in great than in blue tits and that the contribution of competition to variation in these variables is relatively higher in great than in blue tits. 5. Intensity of competition is higher in low quality than in high quality plots for both species, a result not expected from studies in plant communities. 6. Importance of competition varies strongly between plots. It is larger in oak-dominated plots than in mixed deciduous plots. 7. In birds breeding density increases with habitat quality but is limited by territorial behaviour. As a result competition for food is reduced in high quality habitats resulting in a reduction of competition intensity in high quality sites in which birds breed at high densities. 8. It can be predicted that in studies of territorial species density dependent effects on reproduction are more likely to be detected in low quality sites explaining in part differences in results between studies. PMID- 19804540 TI - Defibrillation testing: the need for a definitive trial. PMID- 19804541 TI - Delayed dyssynchronous LV contraction in patients with ischemic cardiomyopathy and narrow QRS complexes is not accompanied by delayed electrical activation: an explanation for lack of CRT success in this group? PMID- 19804542 TI - The impact of catheter ablation on the dynamic function of the left atrium in patients with atrial fibrillation: insights from four-dimensional computed tomographic images. AB - INTRODUCTION: Elucidating the functional properties and remodeling process of the entire left atrium (LA) is important not only for offering the mechanistic insight into atrial fibrillation (AF) but also for assessing the effectiveness of catheter ablation. METHODS: We included 65 patients with paroxysmal AF and 29 controls. Baseline multidetector computed tomography (MDCT) was acquired in all subjects and a follow-up MDCT was available in 48 patients after pulmonary vein and LA ablation. The 3-dimensional images at atrial end-diastole (ED) and end systole (ES) were analyzed. RESULTS: The LA volume (ED: 61.11 +/- 15.94 vs 54.12 +/- 8.94 mL/m(2), P = 0.03; ES: 45.29 +/- 17.64 vs 33.38 +/- 7.78 mL/m(2), P < 0.001) was increased, and ejection fraction (EF) (26.93 +/- 13.40 vs 38.09 +/- 11.62%, P < 0.001) decreased in AF patients as compared to controls. After ablation, the ES LA volume (44.73 +/- 14.93 vs 38.04 +/- 11.51 mL/m(2), P = 0.04) decreased and the LA EF (25.04 +/- 13.13 vs 30.82 +/- 7.85%, P = 0.03) increased in patients without any AF recurrence. The wall motion (WM) analysis of the 18 segments of LA revealed increased motional magnitudes of entire LA except for the anterior roof. In contrast, the volume, EF, and WM of LA remained similar in patients with recurrence. CONCLUSION: Dilated LA with global hypokinesia was noted in AF patients. Improved LA transport function was demonstrated in patients without any recurrence after ablation. However, the anatomic and functional reverse remodeling was not significant in patients with AF recurrence. PMID- 19804543 TI - Characteristics of complex fractionated electrograms in nonpulmonary vein ectopy initiating atrial fibrillation/atrial tachycardia. AB - BACKGROUND: Nonpulmonary vein (PV) ectopy initiating atrial fibrillation (AF)/atrial tachycardia (AT) is not uncommon in patients with AF. The relationship of complex fractionated atrial electrograms (CFAEs) and non-PV ectopy initiating AF/AT has not been assessed. We aimed to characterize the CFAEs in the non-PV ectopy initiating AF/AT. METHODS: Twenty-three patients (age 53 +/- 11 y/o, 19 males) who underwent a stepwise AF ablation with coexisting PV and non PV ectopy initiating AF or AT were included. CFAE mapping was applied before and after the PV isolation in both atria by using a real-time NavX electroanatomic mapping system. A CFAE was defined as a fractionation interval (FI) of less than 120 ms over 8-second duration. A continuous CFAE (mostly, an FI < 50 ms) was defined as electrogram fractionation or repetitive rapid activity lasting for more than 8 seconds. RESULTS: All patients (100%) with non-PV ectopy initiating AF or AT demonstrated corresponding continuous CFAEs at the firing foci. There was no significant difference in the FI among the PV ostial or non-PV atrial ectopy or other atrial CFAEs (54.1 +/- 5.6, 58.3 +/- 11.3, 52.8 +/- 5.8 ms, P = 0.12). Ablation targeting those continuous CFAEs terminated the AF and AT and eliminated the non-PV ectopy in all patients (100%). During a follow-up of 7 months, 22% of the patients had an AF recurrence with PV reconnections. There was no recurrence of any ablated non-PV ectopy during the follow-up. CONCLUSION: The sites of the origin of the non-PV ectopies were at the same location as those of the atrial continuous CFAEs. Those non-PV foci were able to initiate and sustain AF/AT. By limited ablation targeting all atrial continuous CFAEs, the AF could be effectively eliminated. PMID- 19804544 TI - Canadian Registry of ICD Implant Testing procedures (CREDIT): current practice, risks, and costs of intraoperative defibrillation testing. AB - BACKGROUND: There is uncertainty about the proper role of defibrillation testing (DT) at the time of implantable cardioverter defibrillator (ICD) insertion. METHODS: A prospective registry was conducted at 13 sites in Canada between January 2006 and October 2007. OBJECTIVES: To document the details of DT, the reasons for not conducting DT, and the costs and complications associated with DT. RESULTS: DT was conducted at implantation in 230 of 361 patients (64%). DT was more likely to be conducted for new implants compared with impulse generator replacements (71% vs 32%, P = 0.0001), but was similar for primary and secondary prevention indications (64% vs 63%, P = NS). Among patients not having DT, the reason(s) given were: considered unnecessary (44%); considered unsafe, mainly due to persistent atrial fibrillation (37%); lack of an anesthetist (20%); and, patient or physician preference (6%). When performed, DT consisted of a single successful shock > or = 10J below maximum device output in 65% of cases. A 10J safety-margin was met by 97% of patients, requiring system modification in 2.3%. Major perioperative complications occurred in 4.4% of patients having DT versus 6.6% of patients not having DT (P = NS). ICD insertion was $844 more expensive for patients having DT (P = 0.16), largely due to increased costs ($28,017 vs $24,545) among patients having impulse generator replacement (P = 0.02). CONCLUSIONS: DT was not performed in a third of ICD implants, usually due to a perceived lack of need or relative contraindication. PMID- 19804545 TI - Image-integration of intraprocedural rotational angiography-based 3D reconstructions of left atrium and pulmonary veins into electroanatomical mapping: accuracy of a novel modality in atrial fibrillation ablation. AB - INTRODUCTION: Exact visualization of complex left atrial (LA) anatomy is crucial for safety and success rates when performing catheter ablation of atrial fibrillation (AF). The aim of our study was to validate the accuracy of integrating rotational angiography-based 3-dimensional (3D) reconstructions of LA and pulmonary vein (PV) anatomy into an electroanatomical mapping (EAM) system. METHODS AND RESULTS: In 38 patients (62 +/- 8 years, 25 females) undergoing catheter ablation of paroxysmal (n = 19) or persistent (n = 19) AF, intraprocedural rotational angiography of LA and PVs was performed. The subsequent 3D reconstruction and segmentation of LA and PVs was transferred to the EAM system and registered to the EAM. The distances of all EAM points to corresponding points on the LA syngo DynaCT Cardiac surface were calculated. Segmentation of LA with clear visualization of adjacent structures was possible in all patients. Also, the integrated segmentation of the LA was used to guide the encirclement of ipsilateral veins, which resulted in PV isolation in all patients. Integration into the 3D mapping system was achieved with a distance error of 2.2 +/- 0.4 mm when compared with the EAM surface. Subgroups with paroxysmal and persistent AF showed distance errors of 2.3 +/- 0.3 mm and 2.1 +/- 0.4 mm, respectively (P = n.s.). CONCLUSION: Intraprocedural registration of LA and PV anatomy by contrast enhanced rotational angiography was feasible and accurate. There were no differences between patients with paroxysmal or persistent AF. Therefore, integration of rotational angiography-based reconstructions into 3D EAM systems might be helpful to guide catheter ablation for AF. PMID- 19804546 TI - Electrocardiographic findings, device therapies, and comorbidities in octogenarian implantable defibrillator recipients. AB - BACKGROUND: The use of implantable cardioverter-defibrillators (ICDs) in octogenarians is poorly defined. OBJECTIVE: To assess baseline electrocardiographic (ECG) findings, arrhythmia episodes, and development of severe nonarrhythmic illness or death in patients aged >or=80 years at ICD implantation, and to compare them with younger patients. METHODS: Medical records and device interrogations for 199 patients >or=70 years old who underwent ICD implantation were reviewed. Patients were divided into 3 groups based on age at the time of implant: age 70-74 (group 1; 88 patients), age 75-79 (group 2; 67 patients), and age >or=80 (group 3; 44 patients). RESULTS: ECGs: Octogenarians were more likely to have sinus bradycardia (SB) (P = 0.047) and left bundle branch block (LBBB) (P = 0.043) compared to younger patients. There was no difference among groups in the proportion of patients with atrial fibrillation or any degree of AV block. THERAPIES: There was no difference between age groups in any therapy (P = 0.78), appropriate therapy (P = 0.54), or inappropriate therapy (P = 0.21) per patient-year. There was no difference between groups in time-to first therapy of any type (P = 0.71). NONARRHYTHMIC DEATH/MORBIDITY: There was no difference between groups in time to death or serious illness. CONCLUSION: Our study is the first to evaluate in detail the therapies received by octogenarians after ICD placement. The higher incidence of SB and LBBB might influence the number of pacing sites in octogenarian patients. Very elderly patients have similar rates of arrhythmic episodes and development of severe comorbidities as septuagenarians, and they should not be denied ICD implantation based solely on age. PMID- 19804547 TI - Chronic augmentation of the parasympathetic tone to the atrioventricular node: a nonthoracotomy neurostimulation technique for ventricular rate control during atrial fibrillation. AB - INTRODUCTION: The right inferior ganglionated plexus (RIGP) selectively innervates the atrioventricular node. Temporary electrical stimulation of this plexus reduces the ventricular rate during atrial fibrillation (AF). We sought to assess the feasibility of chronic parasympathetic stimulation for ventricular rate control during AF with a nonthoracotomy intracardiac neurostimulation approach. METHODS AND RESULTS: In 9 mongrel dogs, the small endocardial area inside the right atrium, which overlies the RIGP, was identified by 20 Hz stimulation over a guiding catheter with integrated electrodes. Once identified, an active-fixation lead was implanted. The lead was connected to a subcutaneous neurostimulator. An additional dual-chamber pacemaker was implanted for AF induction by rapid atrial pacing and ventricular rate monitoring. Continuous neurostimulation was delivered for 1-2 years to decrease the ventricular rate during AF to a range of 100-140 bpm. Implantation of a neurostimulation lead was achieved within 37 +/- 12 min. The latency of the negative dromotropic response after on/offset or modulation of neurostimulation was <1 s. Continuous neurostimulation was effective and well tolerated during a 1-2 year follow-up with a stimulation voltage <5 V. The neurostimulation effect displayed a chronaxie-rheobase behavior (chronaxie time of 0.07 +/- 0.02 ms for a 50% decrease of the ventricular rate during AF). CONCLUSION: Chronic parasympathetic stimulation can be achieved via a cardiac neurostimulator. The approach is safe, effective, and well tolerated in the long term. The atrioventricular nodal selectivity and the opportunity to adjust the negative dromotropic effect within seconds may represent an advantage over pharmacological rate control. PMID- 19804548 TI - Septal dyskinesia and global left ventricular dysfunction in pediatric Wolff Parkinson-White syndrome with septal accessory pathway. AB - INTRODUCTION: Echocardiographic studies have shown that some patients with Wolff Parkinson-White (WPW) syndrome have myocardial dyskinesia in the segments precociously activated by an accessory pathway (AP). The aim of the present study was to determine the extent to which the AP contributes to global left ventricular (LV) dysfunction. METHODS: Electrophysiological and echocardiographic data from 62 children with WPW (age at diagnosis = 5.9 +/- 4.2 years) were retrospectively analyzed. RESULTS: The left ventricular ejection fraction (LVEF) of patients with septal APs (53 +/- 11%) was significantly lower than that of patients with right (62 +/- 5%) or left (61 +/- 4%) APs (P = 0.001). Compared to patients with normal septal motion (n = 56), patients with septal dyskinesia (n = 6) had a reduced LVEF (61 +/- 4% and 42 +/- 5%, respectively) and an increased LV end diastolic dimension (P < 0.001 for both comparisons). Multivariate analysis identified septal dyskinesia as the only significant risk factor for reduced LVEF. All 6 patients with septal dyskinesia had right septal APs, and a preexcited QRS duration that was longer than that of patients with normal septal motion (140 +/- 18 ms and 113 +/- 32 ms, respectively; P = 0.045). After RFA there were improvements in both intraventricular dyssynchrony (septal-to posterior wall motion delay, from 154 +/- 91 ms to 33 +/- 17 ms) and interventricular septal thinning (from 3.0 +/- 0.5 mm to 5.3 +/- 2.6 mm), and a significant increase in LVEF (from 42 +/- 5% to 67 +/- 8%; P = 0.001). CONCLUSION: The dyskinetic segment activated by a right septal AP in WPW syndrome may lead to ventricular dilation and dysfunction. RFA produced mechanical resynchronization, reverse remodeling, and improvements in LV function. PMID- 19804549 TI - Magnetic resonance imaging-confirmed ablative debulking of the left atrial posterior wall and septum for treatment of persistent atrial fibrillation: rationale and initial experience. AB - INTRODUCTION: Though pulmonary vein (PV) isolation has been widely adopted for treatment of atrial fibrillation (AF), recurrence rates remain unacceptably high with persistent and longstanding AF. As evidence emerges for non-PV substrate changes in the pathogenesis of AF, more extensive ablation strategies need further study. METHODS: We modified our PV antrum isolation procedure to include abatement of posterior and septal wall potentials. We also employed recently described image-processing techniques using delayed-enhancement (DE) MRI to characterize tissue injury patterns 3 months after ablation, to assess whether each PV was encircled with scar, and to assess the impact of these parameters on procedural success. RESULTS: 118 consecutive patients underwent debulking procedure and completed follow-up, of which 86 underwent DE-MRI. The total left atrial (LA) radiofrequency delivery correlated with percent LA scarring by DE-MRI (r = 0.6, P < 0.001). Based on DE patterns, complete encirclement was seen in only 131 of 335 PVs (39.1%). As expected, Cox regression analysis showed a significant relationship between the number of veins encircled by delayed enhancement and clinical success (hazard ratio of 0.62, P = 0.015). Also, progressive quartile increases in postablation posterior and septal wall scarring reduced recurrences rates with a HR of 0.65, P = 0.022 and 0.66, P = 0.026, respectively. CONCLUSION: Pathologic remodeling in the septal and posterior walls of the LA helps form the pathogenic substrate for AF, and these early results suggest that more aggressive treatment of these regions appears to correlate with improved ablation outcomes. Noninvasive imaging to characterize tissue changes after ablation may prove essential to stratifying recurrence risk. PMID- 19804550 TI - Catheter-based cryoablation of postinfarction and idiopathic ventricular tachycardia: initial experience in a selected population. AB - INTRODUCTION: Transvenous cryoablation has proven to be safe and effective for the treatment of supraventricular arrhythmias. The aim of this prospective study was to report the feasibility and safety of catheter-based cryoablation for the treatment of postinfarction and idiopathic ventricular tachycardia (VT). METHODS AND RESULTS: Catheter-based cryoablation was performed in 17 patients (15 men, 58 +/- 18 years). VT occurred after a prior myocardial infarction in 10 and was idiopathic in 7 patients. Cryoablation was performed with a 10-F, 6.5-mm tipped catheter. The ablation site was selected using entrainment mapping techniques for postinfarction VT. The site of the earliest activation time with optimal pace mapping was used for ablation of idiopathic VT. All targeted VTs (12 postinfarction and 7 idiopathic) were acute successfully ablated after a median number of 2 applications of 5 minutes with an average temperature of -82 +/- 4 degrees C. Mean procedure and fluoroscopy times were 204 +/- 52 and 52 +/- 20 minutes for postinfarction VT and 203 +/- 24 and 38 +/- 15 minutes for idiopathic VT. No cryocatheter or cryoenergy complications were observed. After a follow-up of 6 months, 4 of the 10 patients with postinfarction VT had a recurrence. In 1 of the 7 patients with idiopathic VT the index arrhythmia recurred. CONCLUSION: In this small patient population, catheter-based cryoablation of VT was safe and effective. Future studies are needed to evaluate the effect of cryothermy in a larger group of patients, especially those with postinfarction VT. PMID- 19804551 TI - Are MADIT II criteria for implantable cardioverter defibrillator implantation appropriate for Chinese patients? AB - BACKGROUND: MADIT-II demonstrated that prophylactic implantation of an implantable cardioverter-defibrillator (ICD) device prevents sudden cardiac death (SCD) in patients with myocardial infarction (MI) and impaired left ventricular ejection fraction (LVEF). It remains unclear whether the MADIT-II criteria for ICD implantation are appropriate for Chinese patients. METHODS AND RESULTS: We compared the clinical characteristics and outcome for a cohort of consecutive Chinese patients who satisfied MADIT-II criteria for ICD implantation with the original published MADIT-II population. Seventy consecutive patients who satisfied MADIT-II criteria but did not undergo ICD implantation (age: 67 years, male: 77%) were studied. Their baseline demographics were comparable with the original MADIT-II cohort with the exception of a higher incidence of diabetes mellitus. After follow-up of 35 months, most deaths (78%) were due to cardiac causes (72% due to SCD). The 2-year SCD rate (10.0%) was comparable with that of the MADIT-II conventional group (12.1%), but higher than the MADIT-II defibrillator group (4.9%). Similarly, the 2-year non-SCD rate was 3.0%, also comparable with the MADIT-II conventional group (4.6%), but lower than the MADIT II defibrillator group (7.0%). Cox regression analysis revealed that advance NYHA function class (Hazard Ratio [HR]: 3.5, 95% Confidence Interval [CI]: 1.48-8.24, P = 0.004) and the lack of statin therapy (HR: 3.7, 95%CI: 1.35-10.17, P = 0.011) were independent predictors for mortality in the MADIT-II eligible patients. CONCLUSION: Chinese patients who satisfy MADIT-II criteria for ICD implantation are at similar risk of SCD and non-SCD as the original MADIT-II subjects. Implantation of an ICD in Chinese patients is appropriate. PMID- 19804552 TI - Idiopathic left ventricular arrhythmias originating adjacent to the left aortic sinus of valsalva: electrophysiological rationale for the surface electrocardiogram. AB - BACKGROUND: Idiopathic ventricular arrhythmias (VAs) may be amenable to catheter ablation within or adjacent to the left sinus of Valsalva (LSOV). However, features that discriminate these sites have not been defined. The purpose of this study was to determine the electrocardiographic and electrophysiological features of VAs originating within or adjacent to the LSOV. METHODS AND RESULTS: We studied 48 consecutive patients undergoing successful catheter ablation of idiopathic VAs originating from the left coronary cusp (LCC, n = 29), aortomitral continuity (AMC, n = 10) and great cardiac vein or anterior interventricular cardiac vein (Epi, n = 9). A small r wave, or rarely an R wave, was typically observed in lead I during the VAs and pacing in these regions. An S wave in lead V5 or V6 occurred significantly more often during both the VAs and pacing from the AMC than during that from the LCC and Epi (p < 0.05 to 0.0001). For discriminating whether VA origins can be ablated endocardially or epicardially, the maximum deflection index (MDI = the shortest time to the maximum deflection in any precordial lead/QRS duration) was reliable for VAs arising from the AMC (100%), but was less reliable for LCC (73%) and Epi (67%) VAs. In 3 (33%) of the Epi VAs, the site of an excellent pace map was located transmurally opposite to the successful ablation site (LCC = 1 and AMC = 2). CONCLUSIONS: The MDI has limited value for discriminating endocardial from epicardial VA origins in sites adjacent to the LSOV probably due to preferential conduction, intramural VA origins or myocardium in contact with the LCC. PMID- 19804553 TI - Electrophysiological anatomy of typical atrial flutter: the posterior boundary and causes for difficulty with ablation. AB - BACKGROUND: The electrophysiological anatomy of cavotricuspid isthmus-dependent atrial flutter (CVTI-AFL) has not been fully elucidated. METHODS: We studied 602 autopsied human hearts from individuals aged 0 to 103 years. We measured morphological features of the right atrium, including the crista terminalis (CT), pectinate muscles, sub-Eustachian pouch, Thebesian valve (TV), and the coronary sinus (CS) ostium. RESULTS: In adults, the mean right atrium dimensions were 4.7 cm x 4.5 cm x 4.4 cm. Pectinate muscles extended medial to the CT in 54% of hearts. In 19% of hearts, these ended in another ridge termed the second CT. Pectinate muscles extended into the CVTI in 70% of hearts. A sub-Eustachian pouch was present in 16% of hearts, was always located on the septal CVTI, and was more likely when a prominent TV was also present. A TV, present in 62% of all hearts, covered the inferior quadrant of the CS ostium in 9% of these hearts. CONCLUSION: The posterior boundary of the reentrant circuit of CVTI-AFL comprises the Eustachian ridge and CT, but in some patients may also include a second CT. Sub Eustachian pouches on the septal CVTI are strongly associated with a prominent TV. The lateral CVTI can have prominent pectinate muscles. This comprehensive characterization of the electrophysiological anatomy of the reentrant circuit of CVTI-AFL may provide guidance and improve success during difficult ablations. PMID- 19804554 TI - Empowerment and Indigenous Australian health: a synthesis of findings from Family Wellbeing formative research. AB - This paper employs a thematic qualitative analysis to synthesise seven discrete formative evaluation reports of an Indigenous Australian family empowerment programme across four study settings in Australia's Northern Territory and Queensland between 1998 and 2005. The aim of the study, which involved a total of 148 adult and 70 school children participants, is to develop a deeper understanding of the contribution of community empowerment education programmes to improving Indigenous health, beyond the evidence derived from the original discrete micro evaluative studies. Within a context beset by trans-generational grief and despair resulting from colonisation and other discriminatory government policies, across the study sites, the participants demonstrated enhanced capacity to exert greater control over factors shaping their health and wellbeing. Evident in the participants' narratives was a heightened sense of Indigenous and spiritual identity, respect for self and others, enhanced parenting and capacity to deal with substance abuse and violence. Changes at the personal level influenced other individuals and systems over time, highlighting the ecological or multilevel dimensions of empowerment. The study reveals the role of psychosocial empowerment attributes as important foundational resources in helping people engage and benefit from health and other behaviour modification programmes, and take advantage of any reforms made within macro policy environments. A key limitation or challenge in the use of psychosocial empowerment programmes relates to the time and resources required to achieve change at population level. A long-term partnership approach to empowerment research that creatively integrates micro community empowerment initiatives with macro policies and programmes is vital if health gains are to be maximised. PMID- 19804555 TI - Disclosing clinical adverse events to patients: can practice inform policy? AB - OBJECTIVES: To understand patients' and health professionals' experience of Open Disclosure and how practice can inform policy. BACKGROUND: Open Disclosure procedures are being implemented in health services worldwide yet empirical evidence on which to base models of patient-clinician communication and policy development is scant. DESIGN, SETTING AND PARTICIPANTS: A qualitative method was employed using semi-structured open-ended interviews with 154 respondents (20 nursing, 49 medical, 59 clinical/administrative managerial, 3 policy coordinators, 15 patients and 8 family members) in 21 hospitals and health services in four Australian states. RESULTS: Both patients and health professionals were positive about Open Disclosure, although each differed in their assessments of practice effectiveness. We found that five major elements influenced patients' and professionals' experience of openly disclosing adverse events namely: initiating the disclosure, apologizing for the adverse event, taking the patient's perspective, communicating the adverse event and being culturally aware. CONCLUSIONS: Evaluating the impact of Open Disclosure refines policy implementation because it provides an evidence base to inform policy. Health services can use specific properties relating to each of the five Open Disclosure elements identified in this study as training standards and to assess the progress of policy implementation. However, health services must surmount their sensitivity to revealing the extent of error so that research into patient experiences can inform practice and policy development. PMID- 19804556 TI - Implant-supported overdenture in an elderly patient with Huntington's disease. AB - Huntington's disease is a hereditary, progressive, neuro-degenerative disorder characterised by increasingly severe motor impairment, cognitive decline and behavioural manifestations leading to functional disability. Dyskinesia and hyperkinesia of the tongue and the peri-oral musculature make it impossible for the patient to wear a conventional complete denture, despite an adequate alveolar ridge. The present paper reports on a patient with Huntington's disease who was rehabilitated with a mandibular overdenture supported by two endosteal implants. One year follow-up examination showed that the prosthesis was stable and there was considerable improvement in the patient's masticatory function. PMID- 19804557 TI - Experiments on in vivo biofilm formation and in vitro adhesion of Candida species on polysiloxane liners. AB - OBJECTIVES: Microorganisms may colonise polysiloxane soft liners leading to bio deterioration. The aim of this study was to investigate in vitro adhesion and in vivo biofilm formation of Candida species on polysiloxane surfaces. METHODS: The materials used in this study were Molloplast B, GC Reline soft, Mollosil Plus, Silagum Comfort and Palapress Vario. The in vitro retention of clinical isolates of Candida albicans to the relining and denture-base materials by microscopic (scanning electron microscopy, SEM), conventional culturing methods and antimicrobial properties of these materials were studied. Candida found on materials and mucosa following long-term use were identified and quantified, and biofilms covering the surfaces were investigated by SEM. RESULTS: There was a significant decrease in the number of cells attached in vitro to saliva-coated surfaces compared with non-treated surfaces. An oral Candida carriage of 78% was found. Candida albicans, C. glabrata, C. intermedia and C. tropicalis were identified. In vivo biofilm formation on the liners appeared as massive colonisation by microorganisms. CONCLUSIONS: The results of the in vitro experiments suggest that salivary film influences early colonisation of different C. albicans strains. The film layer also minimises the differences among different strains. The Candida carriage of these patients was similar to denture wearing patients without soft liners. PMID- 19804558 TI - Long-term cognitive enrichment affects opioid receptor expression in the amygdala of domestic pigs. AB - Enriching the housing environment by stimuli that challenge both reward and cognitive mechanisms may enhance behavioural experiences and can improve animal welfare, particularly in farm animals. A newly developed experimental feeding system for domestic pigs using food-rewarded learning of discriminatory and instrumental tasks enabled the animals to successfully master a cognitive challenge and to be rewarded ca. 30 times per day with small food portions. Reward-related behaviour is expected to be modulated by endogenous opioid systems. Furthermore, recent evidence supports a role for the amygdala in processing positive affects by stimulus-reward learning. Hence, the present study investigates mRNA expression of cerebral receptors, which are involved in these processes. In an initial step, reverse transcription-polymerase chain reaction (RT-PCR) provided the first evidence that transcripts of three different opioid receptors (MOR, DOR, KOR), as well as the neuropeptide Y 5 receptor (NPY5R), leptin receptor (LEPR) and proopiomelanocortin (POMC), are expressed in both the porcine amygdala and hypothalamus. Using real-time PCR we could show that the expression of two receptors of the opioid system (amygdala: KOR, DOR), in addition to the expression of NPY5R (hypothalamus) in eight enriched housed pigs was markedly downregulated compared to that of conventionally housed and fed pigs. Focusing on opioid receptors in the amygdala, the present study shows that long-term cognitive enrichment acts as a biologically relevant stimulus that causes modifications of gene expression of reward-sensitive cerebral receptors in domestic pigs. PMID- 19804559 TI - Early life stress, MAOA, and gene-environment interactions predict behavioral disinhibition in children. AB - Several, but not all, studies have shown that the monoamine oxidase A functional promoter polymorphism (MAOA-LPR) interacts with childhood adversity to predict adolescent and adult antisocial behavior. However, it is not known whether MAOA LPR interacts with early life (pre-birth-3 years) stressors to influence behavior in prepubertal children. The Avon Longitudinal Study of Parents and Children, UK, is a community-representative cohort study of children followed from pre-birth onwards. The impact of family adversity from pre-birth to age 3 years and stressful life events from 6 months to 7 years on behavioral disinhibition was determined in 7500 girls and boys. Behavioral disinhibition measures were: mother reported hyperactivity and conduct disturbances (Strengths and Difficulties Questionnaire) at ages 4 and 7 years. In both sexes, exposure to family adversity and stressful life events in the first 3 years of life predicted behavioral disinhibition at age 4, persisting until age 7. In girls, MAOA-LPR interacted with stressful life events experienced from 6 months to 3.5 years to influence hyperactivity at ages 4 and 7. In boys, the interaction of MAOA-LPR with stressful life events between 1.5 and 2.5 years predicted hyperactivity at age 7 years. The low activity MAOA-LPR variant was associated with increased hyperactivity in girls and boys exposed to high stress. In contrast, there was no MAOA-LPR interaction with family adversity. In a general population sample of prepubertal children, exposure to common stressors from pre-birth to 3 years predicted behavioral disinhibition, and MAOA-LPR- stressful life event interactions specifically predicted hyperactivity. PMID- 19804560 TI - A novel HLA-Cw*01 variant allele, HLA-Cw*0130. AB - The novel HLA-Cw*0130 variant allele differs from the closest allele Cw*010201 by single nucleotide change at genomic nt 959 T>C (CDS nt 583 T>C, codon 171 TAC>CAC) in exon 3, which causes an amino acid change Tyr171His. PMID- 19804561 TI - A novel HLA-Cw*08 allele, Cw*0822, identified by genomic full-length cloning and sequencing. AB - A novel HLA-Cw*08 allele, Cw*0822, differing from the closest related allele Cw*080101 by one nucleotide exchange at nt2557 (G>A) in exon 6, was identified by genomic full-length cloning and sequencing from a male Chinese donor. PMID- 19804562 TI - Predominant association of HLA-B*2704 with ankylosing spondylitis in Chinese Han patients. AB - The HLA-B27 subtypes have a varied racial and ethnic prevalence throughout the world. However, the association of B27-subtypes with ankylosing spondylitis (AS) in the mainland China is unknown. To determine the association of B27-subtypes with AS in the Mainland Chinese Han population, a total of unrelated 153 patients with AS were enrolled in a large case-control association study, and 1545 unrelated, healthy, ethnically matched blood donors were included as controls. The genotyping of B27 and its subtypes was performed using the polymerase chain reaction with sequence specific primers (PCR-SSP). A total of 130 (84.97%) AS patients and 61 (3.95%) healthy controls were B27 positive. Three B27-subtypes, B*2704, B*2705 and B*2710, were further identified, of which both B*2704 and B*2705 were strongly AS associated. B*2710 was only detected in one AS patient and two other healthy controls. Considering only B27-positive cases and controls, a statistically different frequency of B27-subtypes was observed, with an over representation of B*2704 (P = 0.018). B*2704 was clearly more strongly associated than B*2705 with AS [odds ratio (OR ) = 2.4, P = 0.011]. Furthermore, a combined analysis including three previous studies of B27-subtype distributions in Chinese AS cases confirmed the stronger association of B*2704 with AS than B*2705 (OR = 2.5, P = 0.00094). PMID- 19804563 TI - Incidence of humoral sensitization in HLA partially mismatched hematopoietic stem cell transplantation. AB - As part of the 15th International Histocompatibility and Immunogenetics Workshop (IHIWS), seven centers participated in a collaborative project to determine whether any significant humoral sensitization occurred post-transplant among recipients of HLA partially mismatched hematopoietic cell transplants (HCTs). A total of 140 donor/recipient pairs were enrolled with a total of 367 pre-and post transplant sera analyzed. The majority of the samples (69.1%) were obtained within 30-90 days post-HCT. HLA-specific antibodies were defined using single antigen bead assays on a Luminex platform with a positive cutoff value of 1000 normalized median fluorescence intensity (MFI). There was an overall incidence of post-HCT sensitization toward donor HLA mismatches of 5.7%; however, all cases were among recipients of one HLA haplotype-mismatched grafts under nonmyeloablative, pre-transplant conditioning. Among the one haplotype-mismatched recipients, 15.7% (8/51) developed donor HLA-specific antibodies and 29.4% also had antibodies directed toward third party HLA antigens. Among the donor-specific antibodies, 9.8% were directed toward HLA class I antigens; 7.8% were against class II antigens; and 2.0% had both class I and II specificity. The relative strength of post-transplant antibodies was low with no significant difference in the mean maximum MFI values between third party and donor-specific antibodies. Because only a small number (10.2%) of the post-transplant samples were obtained 180 days or more post-HCT, longer term study is needed to evaluate any clinical relevance of these low-to-moderate levels of donor-specific antibody in one haplotype-mismatched recipients, as well as to determine whether any other antibodies occur at later times. PMID- 19804564 TI - Genomic full-length sequence of HLA-Cw*140201, identified by cloning and sequencing. AB - Genomic full-length sequence of HLA-Cw*140201, differing from Cw*1403 by four nucleotide exchanges in exon 2 and intron 5, was identified by cloning and sequencing from a male Chinese donor. PMID- 19804565 TI - Different requirements for early and late phases of azurophilic granule-phagosome fusion. AB - Phagocytosis and killing of microorganisms are complex processes that involve tightly regulated membrane traffic events. Because many signaling molecules associate with membrane rafts and because these structures can be found on azurophilic granules, we decided to investigate raft recruitment and the signaling requirements for azurophilic granule secretion during phagosome maturation. At the site of phagocytosis of immunoglobulin G-opsonized prey in human neutrophils, we found that early secretion of azurophilic granules was both raft- and calcium-dependent. Subsequently, rafts at the phagocytic site were internalized with the prey. At the fully formed phagosome, the fusion of azurophilic granules was no longer dependent on rafts or calcium. These findings were found to be true also when using Streptococcus pyogenes bacteria as prey, and depletion of calcium affected the kinetics of bacterial intracellular survival. These findings suggest that the mechanisms for delivery of azurophilic content to nascent and sealed phagosomes, respectively, differ in their dependence on calcium and membrane rafts. PMID- 19804566 TI - Pivotal role of translokin/CEP57 in the unconventional secretion versus nuclear translocation of FGF2. AB - Intracellular trafficking of fibroblast growth factor 2 (FGF2) exhibits two unusual features: (i) it is secreted despite the lack of signal peptide and (ii) it can translocate to the nucleus after interaction with high- and low-affinity receptors on the cell surface, although it does not possess any classical nuclear localization signal. This nuclear translocation constitutes an important part of the response to the growth factor. Previously, we identified Translokin/CEP57, an FGF2 binding partner, as an intracellular mediator of FGF2 trafficking, which is essential for the nuclear translocation of the growth factor. Here, we report the identification of four Translokin partners: sorting nexin 6, Ran-binding protein M and the kinesins KIF3A and KIF3B. These proteins, through their interaction with Translokin, are involved in two exclusive complexes allowing the bidirectional trafficking of FGF2. Thus, Translokin plays a pivotal role in this original mechanism. In addition, we show that FGF2 secretion is regulated by a negative loop, retro-controlled by FGF receptor and involving FGF2 itself. PMID- 19804567 TI - The practice of reporting transfusion-related acute lung injury: a national survey among clinical and preclinical disciplines. AB - BACKGROUND: Transfusion-related acute lung injury (TRALI) is hypothesized to be a "two-hit" entity, in which an inflammatory condition (e.g., sepsis) predisposes to TRALI. TRALI is a clinical diagnosis. Disciplines involved in managing TRALI may differ in decision-making on the reporting of TRALI. STUDY DESIGN AND METHODS: A survey was conducted among critical care physicians, hematologists, hemovigilance workers, and transfusion medicine physicians, using case vignettes and a questionnaire. The vignettes varied in patient- and blood product-related factors that may influence the decision to report a TRALI case. Multiple linear regression analysis was performed. A positive beta-coefficient is in favor of reporting. RESULTS: Ninety-two questionnaires were returned (response rate, 68%). For all disciplines, preferences in favor of reporting TRALI were onset of symptoms within 1 hour (beta = 0.4), after transfusion of a single unit of FFP (beta = 0.5), and in the absence of acute lung injury before transfusion (beta = 1.3). An admission diagnosis of sepsis was a negative preference (beta = -0.3). Massive transfusion (6 RBC plus 4 FFP units) was a negative preference for transfusion medicine physicians (beta = -0.3), but a positive preference for the other disciplines. The questionnaire revealed that massive transfusion and the age of blood products were considered relatively more important reasons to report TRALI by critical care physicians compared to the other disciplines (p < 0.05). CONCLUSION: A pretransfusion inflammatory condition is a reason to withhold from reporting of a suspected TRALI case. Disciplines involved in managing TRALI differ in decision-making of reporting TRALI, which may contribute to variance in incidence. PMID- 19804568 TI - Increased number of coagulation products in relationship to red blood cell products transfused improves mortality in trauma patients. AB - BACKGROUND: Recent data from military and civilian centers suggest that mortality is decreased in massive transfusion patients by increasing the transfusion ratio of plasma and platelet (PLT) products, and fibrinogen in relationship to red blood cell (RBC) products during damage control resuscitation and surgery. This study investigates the relationship of plasma:RBC, PLT:RBC, and cryoprecipitate:RBC transfusion ratios to mortality in massively transfused patients at a civilian Level 1 trauma center. STUDY DESIGN AND METHODS: Demographic, laboratory, transfusion, and outcome data were collected prospectively from February 1, 2007, to January 31, 2009, and retrospectively from February 1, 2005, to January 31, 2007, on all injured patients who underwent massive transfusion (defined as >or=10 RBC products within 24 hr). Mortality was analyzed in relation to the plasma:RBC, PLT:RBC, and cryoprecipitate:RBC transfusion ratios using both univariate and multivariate analyses. RESULTS: A total of 214 patients received massive transfusion secondary to traumatic injury. High versus low transfusion ratios were associated with improved 30-day survival: plasma:RBC 59% versus 44%, p = 0.03; PLT:RBC 63% versus 33%, p < 0.01; and cryoprecipitate:RBC 66% versus 41%, p < 0.01. By multivariable stepwise logistic regression analysis, increased plasma:RBC (p = 0.02) and PLT:RBC (p = 0.02), and decreased age (p = 0.02), ISS (p < 0.01) and total RBCs (p = 0.03) were statistically associated with improved 30-day survival. CONCLUSIONS: In the civilian setting, plasma, PLT, and cryoprecipitate products significantly increased 30-day survival in trauma patients. Future prospective randomized clinical trials are required to determine the optimal transfusion ratios. PMID- 19804569 TI - Prospective monitoring of plasma and platelet transfusions in a large teaching hospital results in significant cost reduction. AB - BACKGROUND: Plasma and platelets (PLTs) are often transfused to correct mild to moderately abnormal laboratory values. Our objective was to reduce unnecessary plasma and PLT transfusions to nonbleeding patients by prospective triage and education of end users in evidence-based hemostasis and transfusion medicine practices. STUDY DESIGN AND METHODS: Using the Parkland Memorial Hospital's transfusion service and admission database as the data source, this study comprises the comparison of transfusion data on plasma and PLT use between pre- (2000-2002) and posttriage (2003-2006) periods. Yearly transfusion and wastage data on red blood cells (RBCs), plasma, and PLTs and yearly hospital admissions, trauma visits, and surgical procedures were extracted retrospectively for the study. RESULTS: The study revealed that implementation of triage resulted in a significant reduction of plasma (60%) and PLT (25%) transfusions, saving more than $3,000,000 over 4 years. CONCLUSIONS: Prospective triage and evidence-based transfusion practice education reduced unnecessary plasma and PLT transfusions and health care costs. PMID- 19804570 TI - Improved plasma removal efficiency for therapeutic plasma exchange using a new apheresis platform. AB - BACKGROUND: The Spectra Optia (SPO; CaridianBCT) is a new apheresis device based on the COBE Spectra (CSP; CaridianBCT) platform. This study was designed to evaluate the safety and efficiency of the SPO in comparison to the predicate CSP device. STUDY DESIGN AND METHODS: Twenty patients were recruited for a randomized, nonblinded, paired (crossover) clinical trial comparing the SPO to the CSP (pivotal trial). The primary outcome measure was plasma removal efficiency (PRE); secondary outcomes included platelet (PLT) content and hemolysis in the waste plasma, changes in patient cellular counts, patient coagulation and complement cascade activation, accuracy of machine fluid balance measurement, and review of significant adverse events (SAEs). RESULTS: Overall SPO demonstrated 87% PRE with 1.0% PLT loss; these variables were statistically different from CSP (79 and 3.0%, respectively). The accuracy of anticoagulant usage, plasma removal, and fluid replacement as measured by the SPO fluid pumps was 97% or more; fluid balance was within 2% of the measured value. After apheresis there were no statistical changes in patient cellular counts with respect to the initial values. Patient d-dimer and prothrombin fragment 1.2 assays showed no activation of the coagulation system with either device. Measurement of patient C3a, C5a, and plasma free hemoglobin showed no significant differences between the SPO and the CSP. No SAEs were reported. CONCLUSION: The SPO has improved performance characteristics over the CSP. Based on our results, the SPO is acceptable for use in therapeutic plasma exchange programs. PMID- 19804571 TI - A single assay for multiple storage-sensitive red blood cell characteristics by means of infrared spectroscopy. AB - BACKGROUND: To maintain a high quality of red blood cells (RBCs), RBC characteristics must be followed during storage under blood bank conditions. By means of infrared (IR) spectroscopy, several characteristics can be measured simultaneously. STUDY DESIGN AND METHODS: IR spectra were acquired for samples from RBCs that were collected and stored according to Dutch blood bank procedures for a period of up to 50 days. Spectra of the soluble cell components were acquired separately after hypotonic lysis of the cells, followed by centrifugation. Characteristic vibrational bands were analyzed with respect to storage time-dependent changes in peak position and in intensity. RESULTS: A decrease in corresponding peak intensities indicates that RBCs lose protein and lipid during storage. Changes in protein secondary structure during storage are largely confined to integral membrane proteins and membrane-associated proteins. A concurrent decrease in lipid packing density probably reflects the gradual change in cellular shape from discoidal to globular. By integration over a narrow range, storage-dependent changes in intracellular adenosine triphosphate (ATP) and glucose levels could be estimated. ATP levels decrease during storage, but stay above the required 75% of the initial level after 35 days of storage. Glucose concentrations stay well above 5 mmol/L over the entire storage period. CONCLUSION: IR spectroscopy is a promising technique to follow structural and metabolic changes in RBCs during storage under blood bank conditions. Several variables can be determined rapidly in a single measurement. PMID- 19804572 TI - Many genetically defined ABO subgroups exhibit characteristic flow cytometric patterns. AB - BACKGROUND: A flow cytometric method for detection of low levels of A/B antigen had been developed previously in our laboratory. The aim of this study was to investigate if this approach could be utilized to characterize different ABO subgroups and constitute a useful tool in a reference laboratory. STUDY DESIGN AND METHODS: Blood samples causing ABO discrepancies (n = 94) by routine serology were further analyzed by ABO genotyping and flow cytometry. To verify the specificity of the monoclonal anti-A and -B reagents and to establish normal flow cytometric patterns, samples from 80 blood donors with common phenotypes were also assessed. RESULTS: Distinguishable flow cytometric patterns were detected for several subgroups but were more apparent for A(weak) (n = 80) samples than B(weak) (n = 14). Two subgroups, A(finn) (n = 11) and A(3) (n = 10) displayed diagnostic features and were used to establish reproduciblity over time and between donors. In general, the consistency within subgroups was remarkable. The allelic enhancement phenomenon was clearly visualized among A(x) samples (n = 10) where different alleles in trans resulted in high, low, or no A antigen expression. Nonsubgroup samples including O/A and O/B chimeras or A(h) and B(h) para-Bombay phenotypes displayed clearly distinguishable histograms. Samples from pregnant women (n = 10) displayed acquired A antigen loss, apparently accentuated during the third trimester. CONCLUSIONS: Genetically defined ABO subgroups and other anomalous phenotypes displayed flow cytometric profiles that may contribute valuable information to the investigation of ABO discrepancies. We conclude that the presented assay may complement traditional serology and genetic analysis in the reference laboratory setting. PMID- 19804573 TI - Delayed adverse reactions to blood donation. AB - BACKGROUND: Blood donation is safe, but a small proportion of donors have delayed and/or off-site reactions that have the potential to lead to serious injury. This retrospective study sought to identify risk factors for delayed reactions (DRs). STUDY DESIGN AND METHODS: The records of 793,293 allogeneic whole blood and apheresis donations in 2007 were assessed for vasovagal reactions. Donor demographic, biometric, and clinical measurements were captured. Incidents related to needle insertion and mild reactions were excluded. Based on the reaction onset time relative to the procedure end time, reactions were classified as delayed (>15 min) or immediate (28 days from injury to injury free). Each team sustained a mean of 3.4 hamstring injuries per season, with a mean of 21.5 days missed per injury (range 3-136; median 16 days per injury). The seasonal distribution showed an accumulation of injuries in the first 2 months after a 3.5-month mid-season winter break. PMID- 19804576 TI - Secular trends in physical fitness in Danish adolescents. AB - It is important to monitor changes over time in aerobic fitness and obesity in order to target interventions when health deteriorates. We analyzed data from three population studies of adolescents carried out in 1983, 1997, and 2003. Participants were 1050 adolescents from the whole country of Denmark in 1983, 410 in 1997, and 418 in 2003, the two latter cohorts from the City of Odense. VO(2max) was estimated from maximal power output (MPO) in a cycle test with progressively increasing workload. Estimated VO(2max) for boys and girls was 52 and 41 mL/min/kg, respectively, both in 1983, 1997, and 2003 with no difference between the three cohorts. However, body mass index (BMI) increased 10% in the upper decentile of the distribution since 1983. MPO decreased over time, but validation studies showed that this was not due to decreased VO(2max) . The cohort from 1983 was tested twice by school teachers and experienced scientists, and the scientists found higher MPO than school teachers in the same subjects, which emphasize the importance of good validation studies. PMID- 19804577 TI - Psychometric properties of the Belgian coach version of the coach-athlete relationship questionnaire (CART-Q). AB - The study examined the psychometric properties of the Belgian coach version of the Coach-Athlete Relationship Questionnaire (CART-Q). The questionnaire includes three dimensions (Closeness, Commitment, and Complementarity) in a model that intends to measure the quality of the coach-athlete relationship. Belgian coaches (n=144) of athletes who performed at various competition levels in such sports as football, basketball, and volleyball responded to the CART-Q and to the Leadership Scale for Sport (LSS). A confirmatory factor analysis proved to be slightly more satisfactory for a three-order factor model, compared with a hierarchical first-order factor model. The three factors showed acceptable internal consistency scores. Moreover, functional associations between the three factors and coach leadership behaviors were found offering support to the instrument's concurrent validity. The findings support previous validation studies and verify the psychometric properties of the CART-Q applied to Belgian coaches of team sports. PMID- 19804578 TI - Retraining motor control of abdominal muscles among elite cricketers with low back pain. AB - The purpose of this study was to document the effect of a staged stabilization training program on the motor control of the anterolateral abdominal muscles in elite cricketers with and without low back pain (LBP). Changes in the cross sectional area of the trunk, the thickness of the internal oblique and transversus abdominis (TrA) muscles and the shortening of the TrA muscle in response to an abdominal drawing-in task were measured at the start and completion of a 13-week cricket training camp. Measures were performed using ultrasound imaging and magnetic resonance imaging. Participants from the group with LBP underwent a stabilization training program that involved performing voluntary contractions of the multifidus, TrA and pelvic floor muscles, while receiving feedback from ultrasound imaging. By the end of the training camp, the motor control of cricketers with LBP who received the stabilization training improved and was similar to that of the cricketers without LBP. PMID- 19804579 TI - The role of systemic inflammation in age-related muscle weakness and wasting. AB - Ageing is associated with a slow, but progressive muscle weakness, which is largely attributable to muscle wasting. A diminished function of satellite cells at old age may hamper preservation and repair from (contraction)-induced injury and contribute to the age-related muscle wasting. Satellite cell function may be affected by circulating factors, as muscle regeneration in old mice sharing the circulation of young mice is not impaired. Chronic low-grade systemic inflammation in old organisms may be that environmental factor. Indeed, the inflammatory cytokine tumor necrosis factor-alpha (TNFalpha) negatively affects the muscle regenerating capacity. TNFalpha destabilizes MyoD, a muscle-specific transcription factor involved in satellite cell proliferation and differentiation, and induces apoptosis of satellite cells, particularly at old age. Here it is proposed that some of these effects are mediated by TNFalpha induced expression of inhibitors of differentiation proteins. Yet, the increase in TNFalpha during the normal inflammatory response helps, rather than impairs, the repair process. This apparent contradiction may be resolved by the fact that the effects of TNFalpha are concentration and time dependent. Thus, the negative effect of systemic inflammation on muscle strength at old age may only become apparent when it exceeds a certain threshold and persists for a prolonged period. PMID- 19804580 TI - Comparison of changes in markers of muscle damage induced by eccentric exercise and ischemia/reperfusion. AB - To examine the effects of eccentric exercise (EE) and ischemia/reperfusion (I/R) on the markers of muscle damage, 72 rats were randomly assigned to the EE group, I/R group and control group (C), respectively. The rats in EE ran downhill on a treadmill with a 16 degrees inclination at a constant speed for 90 min, and the rats in the I/R group underwent 90 min of four-limb ischemia, followed by 24, 48 and 72 h of reperfusion. Blood and tissue samples were collected immediately, 24, 48 and 72 h after exercise or reperfusion. Quantitative analyses showed that the I/R group had a significantly larger mitochondrial volume at 24 h after reperfusion compared with the C, and there were more disrupted Z-lines in the EE group and more disrupted mitochondria in the I/R group at 24 h after exercise or reperfusion. When compared with the C, a significantly lower total antioxidant capacity and higher interleukin-6 value were observed after exercise or reperfusion. Our data suggest that although EE and I/R result in some similar changes in the muscle damage markers, there are still some differences. The EE- and I/R-induced muscle damage may be due to different mechanisms. PMID- 19804581 TI - Maximal strength training improves walking performance in peripheral arterial disease patients. AB - Peripheral arterial disease (PAD) patients have reduced muscle strength and impaired walking ability. The aim of this study was to examine the effects of maximal strength training (MST) on walking economy and walking performance in PAD patients. Ten patients with mild to moderate-severe claudication, classified as Fontaine stage II PAD and with functional limitations from intermittent claudication were recruited and went through an 8-week control period followed by an 8-week, three times a week, MST period. The patients performed four sets of five repetitions dynamic leg press with emphasis on maximal mobilization of force in the concentric action and with a progressive adjusted intensity corresponding to 85-90% of one repetition maximum (1 RM). After the MST period, leg press 1 RM significantly increased by 35.0 +/- 10.8 kg (31.3%). Dynamic rate of force development, measured on a force plate installed on the leg press, increased by 1424 +/- 1217 N/s (102.7%). The strength improvements led to a significant increase in walking economy of 9.7% when walking horizontally, and to a significant increase in walking performance of 13.6% measured on an incremental treadmill test to exhaustion. No changes were apparent after the control period. No changes in body mass or peak oxygen uptake were observed. MST increases strength in Fontaine stage II PAD patients and leads to improved walking economy. These results suggest that application of MST could accompany aerobic endurance training as a part of the treatment of PAD patients with mild to moderate-severe claudication. PMID- 19804582 TI - Evidence for low muscle capillary supply as a pathogenic factor in chronic compartment syndrome. AB - There is a paucity of data regarding the pathogenesis of chronic exertional compartment syndrome (CECS), its consequences for the muscles and the effects of treatment with fasciotomy. We analyzed biopsies from the tibialis anterior muscle, from nine patients, obtained during a decompressing fasciotomy and during follow-up 1 year later. Control biopsies were obtained from nine normal subjects. Muscle capillarity, fiber-type composition and fiber area were analyzed with enzyme- and immunohistochemistry and morphometry. At baseline, CECS patients had lower capillary density (273 vs 378 capillaries/mm(2) , P=0.008), lower number of capillaries around muscle fibers (4.5 vs 5.7, P=0.004) and lower number of capillaries in relation to the muscle fiber area (1.1 vs 1.5, P=0.01) compared with normal controls. The fiber-type composition and fiber area did not differ, but focal signs of neuromuscular damage were observed in the CECS samples. At 1 year follow-up after fasciotomy, the fiber area and the number of fibers containing developmental myosin heavy chains were increased, but no enhancement of the capillary network was detected. Thus, morphologically, patients with CECS seemed to have reduced microcirculation capacity. Fasciotomy appeared to trigger a regenerative response in the muscle, however, without any increase in the capillary bed. PMID- 19804583 TI - Donor-derived aspergillosis from use of a solid organ recipient as a multiorgan donor. AB - The growing need for organs and the scarcity of donors has resulted in an increased use of extended criteria donors. We report a case where a recipient of a cardiac graft was used as an organ donor. Death of the recipient occurred 9 days after transplantation and was attributed to presumed cerebral hemorrhage, which post mortem was diagnosed as invasive aspergillosis of the brain. One recipient of a kidney transplant lost the graft due to infection with Aspergillus fumigatus, whereas prompt initiation of therapy successfully prevented disseminated aspergillosis in the other recipients. Despite the pressure to extend the use of organs by lowering the acceptance criteria, organs should only be accepted if the cause of death of the donors is unequivocally explained. PMID- 19804584 TI - Factors and outcome in BK virus nephropathy in a Hispanic kidney transplant population. AB - BK virus nephropathy (BKVN) is an increasingly recognized cause of kidney allograft loss and is thought to be related to the newer, more potent immunosuppressive agents. Conflicting information has been reported on risk factors for BK infection. PURPOSE: To determine incidence, associated factors, and outcome of BKVN in our kidney transplant population in order to improve identification and management. METHODS: Kidney transplants from January 2000 to December 2005 were retrospectively reviewed. Data were collected for patients with biopsy-proven BKVN including age, sex, body mass index (BMI), etiology of renal failure, other medical diseases, donor type, surgical complications, rejection and infection, time to diagnosis, induction, immunosuppressive and antiviral therapy, and clinical outcome. A control group of patients matched for sex, age, type of graft, etiology of kidney disease, and BMI, was established for comparison. RESULTS: STUDY GROUP: During this period, 20 (4%) of 497 transplanted patients were diagnosed with BKVN. Thirteen (65%) were males, 8 (40%) were young adults (ages 21-40), and 18 (90%) received grafts from cadaveric donors (P=0.05). Twelve (60%) had hypertensive renal disease, 2 (10%) also had diabetes, and 16 (80%) had a BMI >25 (P=0.01). Lymphoceles occurred in 5 patients (25%). Mean creatinine level at diagnosis was 2.7 mg/dL and mean time to diagnosis was 23 months. Ten patients (50%) had leukopenia at or within a year before biopsy (P=0.001). Viruses other than BK occurred in 9 patients: varicella zoster virus in 3, cytomegalovirus in 2, herpes simplex virus in 1, molluscum contagiosum in 1, Epstein-Barr virus in 1, and human papillomavirus in 1. Eighteen patients (90%) had related rejection (P= 0.001) and 4 (20%) suffered allograft loss (P= 0.001). Basiliximab (living donors) and anti-thymocyte globulin (cadaver donors) were given for induction. All patients were on triple therapy; 15 on prednisone and sirolimus, with either tacrolimus in 8, cyclosporine in 4, mycophenolate in 1, or mycophenolate and tacrolimus in 2. The other 5 received prednisone with tacrolimus and mycophenolate. Graft loss occurred in 2 patients on tacrolimus and mycophenolate, 1 patient on tacrolimus and sirolimus, and 1 patient on cyclosporine and sirolimus. Immunosuppression was decreased in all patients. Two were given cidofovir for 6 months and had stable creatinine levels at the end of the study. Records were reviewed until April 2007. There were no deaths in this cohort. CONTROL GROUP: The number of rejections experienced by patients with BKV was much higher (P<0.0001), but the rate of graft loss was similar between the 2 groups (P=0.19). Viral co-infection was more frequent in patients with BKV (P=0.04). No episodes of leukopenia were reported for any of the patients in the control group (P=0.001). Immunosuppression with tacrolimus and sirolimus was more frequent in the BKV group, but this was not statistically significant (P=0.18, 0.28, respectively). The number of lymphoceles was larger in patients with BKV, but the difference was not statistically significant (P=0.35). CONCLUSION: BKVN is present in our transplant population and results in a high rate of allograft rejection with varying rates of graft loss. Associated factors were deceased donor and immunosuppression with potent agents, particularly tacrolimus and sirolimus. We also found a higher frequency of obesity, viral co-infection, and leukopenia. Routine screening and timely biopsy could prove cost-effective and significantly reduce morbidity. PMID- 19804585 TI - Increased frequency of regulatory T cells and selection of highly potent CD62L+ cells during treatment of human lung transplant recipients with rapamycin. AB - The currently available immunosuppressive agents applied in human transplantation medicine are highly potent in the protection from acute allograft rejection. However, long-term allograft survival is still poor as these drugs fail to sufficiently prevent chronic allograft rejection. Naturally occurring regulatory T cells have been postulated as the key players to establish long-lasting transplantation tolerance. Thus, the development of immunosuppressive regimens which shift the pathological balance of cytopathic versus regulatory T cells of human allograft recipients towards a protective T-cell composition is a promising approach to overcome limitations of current transplantation medicine. Thirty three patients that received rapamycin (RPM) or calcineurin inhibitor treatment following lung transplantation were included and their T-cell compartments analysed. Twelve healthy volunteers without history of lung disease served as controls. In this article, we show that treatment of human lung transplant recipients with RPM is associated with an increased frequency of regulatory T cells, as compared with treatment with calcineurin inhibitors or to healthy controls. Moreover, regulatory T cells during treatment with RPM were CD62Lhigh, a phenotype that displayed an enhanced immunosuppressive capacity ex vivo. Our data support the use of RPM in human lung transplant recipients and undertaking of further prospective studies evaluating its impact on allograft and patient survival. PMID- 19804586 TI - Evidence review of technology and dietary assessment. AB - SIGNIFICANCE: Diets high in fruit and vegetable consumption are associated with a decrease in chronic diseases. Dietary factors are linked to 4 of the 10 leading noncommunicable causes of death: cardiovascular disease, some cancers, stroke, and type 2 diabetes. Accurately measuring dietary patterns has many challenges. Dietary intake measurement has traditionally relied on self-report instruments such as 24-hour recall, food record, and food frequency questionnaires to record consumption history. These methods have inherent limitations in detecting small but important changes in fruit and vegetable consumption patterns. Promising advances in technology have made more sophisticated techniques for recording dietary intake possible. Computers and Web-based programs, handheld personal digital assistants with cameras and telephone cards, smart phones, cameras, and video recorders options may reduce the burden of recording what has been consumed. Furthermore, technology-based methods of dietary assessment may provide a higher degree of reliability and validity in visually determining fruit and vegetable consumption, and additional study is warranted. PURPOSE: The purpose of this article is to present a review of the evidence on the effectiveness of technology-based methods for dietary assessment, which included fruit and vegetable consumption. APPROACH: One hundred and eighty-seven articles published between 1998 and 2008 were initially identified. Fifteen met the study inclusion criteria and were evaluated by an interdisciplinary team using the Stetler Strength of Evidence Scale. RESULTS: Six technology-based methods for dietary assessment were identified. Findings from validity and reliability testing of technology-based methods are encouraging and need replication. Clinically important features offered through technology may reduce reporting burden and offer behavioral feedback to users. Methodologically sound, empirical research into using technology-based application for dietary assessment in a variety of populations of sufficient size is needed. PMID- 19804587 TI - Transformative learning and research utilization in nursing practice: a missing link? AB - BACKGROUND: Poor or inconsistent research utilization into clinical practice is a recurrent theme across study contexts, rendering leaders disillusioned with how best to foster the uptake of research into nursing practice. This makes it imperative to look to new approaches. Research utilization involves a learning process engaging attitudes, beliefs, and behaviors; yet, this is often overlooked in approaches and models used to facilitate research use. This oversight may offer some explanation to the limited progress in research utilization to date. Transformation Theory offers an explanatory theory and specific strategies (critical reflection and critical discourse) to explore attitudes, beliefs, and behaviors so that they are understood, validated, and can better guide actions. AIM: The purpose of this article was to explore what Transformation Theory can contribute to research utilization initiatives in nursing practice. APPROACH: Transformation Theory and transformative learning strategies are discussed and critically analyzed in consideration of their potential roles in fostering research utilization in clinical nursing practice. ISSUES AND CONCLUSIONS: (1) Research utilization is a learning process that involves knowledge, skills, feelings, attitudes, and beliefs. (2) Transformative learning strategies of critical reflection and discourse can facilitate insight into experiences, finding shared meanings among groups of people, and understanding/validating beliefs, attitudes, and feelings so they can more consciously guide future actions. This dimension is frequently neglected in research utilization efforts. (3) In combination with research utilization theories, Transformation Theory may be a missing link to make research utilization initiatives more effective in rendering and sustaining nursing practice change, thus enhancing client care and well-being. (4) Research and further consideration are both warranted and needed. PMID- 19804588 TI - Pilot study to test the use of a mobile device in the clinical setting to access evidence-based practice resources. AB - AIM: To test the feasibility and acceptability of the use of a mobile device to access evidence-based practice (EBP) resources in the clinical setting. METHODS: A pretest/posttest design was used with a convenience sample of 2nd- and 3rd-year preregistration undergraduate health care students in the United Kingdom. Questionnaires were used to measure (1) feasibility and acceptability of the mobile device and (2) perceptions of the development of EBP knowledge and skills. The study took place during the students' clinical practice and involved two meetings at the beginning and end of the placement period. A Web page was developed to support the process and provide links to key EBP resources. RESULTS: Nineteen undergraduate physiotherapy and nursing students took part in the study. The main findings indicated a generally low level of utilisation of the mobile device in the clinical setting, primarily due to practical difficulties associated with accessing the Internet and the small size of the screen. Consequently, the majority of the students used personal computers (PCs) to access EBP resources. Through the process, students reported improvements in their knowledge and skills in relation to EBP and the appraisal of clinical guidelines. CONCLUSIONS: Students were able to complete the EBP activity using either the mobile device or PC and reported improvements in their knowledge and skills in relation to EBP and the appraisal of clinical guidelines. Findings suggest that for undergraduate health care students, rapid access to online evidence in the clinical environment is not necessarily essential for the integration of the EBP process into practice, or for the development of EBP knowledge and skills, provided there is easy access to such evidence at some point during the placement period. PMID- 19804589 TI - Building a learning collaborative. AB - BACKGROUND: Development and dissemination of evidence-based practice (EBP) guidelines are important first steps, but without active translation efforts, they do little to promote knowledge uptake by direct care providers. In order to close the gap between discovery and use of knowledge, efforts should focus on methods to speed translation of research findings into practice. AIM: To facilitate this process, the National Nursing Practice Network (NNPN) was established to provide participating health care organizations with access to resources, information, opportunities for learning and knowledge transfer, and legitimacy and credibility with internal and external stakeholders. APPROACH: Designed as a practice collaborative learning network, the NNPN uses an interactive learning approach by valuing and learning from the experiences of members and encouraging active participation through conferences, teleconferencing, and use of an interactive member Web site. Use of a network of acute care sites to increase the uptake of EBP is based on social network theory; the NNPN is a network of acute care hospitals through which data, information, knowledge, practices, and behaviors flow. DISCUSSION AND CONCLUSION: The use of a collaborative nursing network provides an innovative way to address the difficulty of implementing and sustaining EBP in organizations and also for maximizing resources and knowledge. Early evaluation data are promising. This approach is offered as one possible method of moving use of EBP in nursing forward. PMID- 19804590 TI - Human eyelash characterization. AB - BACKGROUND: Few biological data on human eyelash follicles have been reported in the literature. OBJECTIVES: To characterize eyelash follicle growth, cycle and morphology, and further investigate the biological mechanisms that determine eyelash length, curl and pigmentation, compared with scalp hair follicle. METHODS: Twenty-nine caucasian female volunteers aged between 26 and 60 years were enrolled in the study to provide eyelashes. Four of these volunteers were followed weekly for 9 months to characterize their eyelash cycle. Eyelash length and time of renewal were measured using a high-resolution camera and image analysis. Immunohistological study of the bulbs were performed on eyelid biopsies from 17 patients requiring block excision for ectropion repair. RESULTS: The calculated durations of anagen phase and complete cycle of the eyelashes were 34 + or - 9 and 90 + or - 5 days, respectively. Eyelash follicle growth rate was quite variable, with an average rate of 0.12 + or - 0.05 mm daily. Eyelash follicle morphology was very close to that of the scalp hair follicle, but some remarkable differences were noticed. For example, the K19-positive epithelial stem cell population was spread all along the follicle and not split into two reservoirs as seen in scalp hair follicles. Some asymmetry was detected in HSPG and CSPG, as well as K38 (formerly Ha8) and K82 (formerly Hb2) distribution, similar to that observed in curly hair. Finally, dopachrome tautomerase was found expressed in eyelash follicle melanocytes, while it was strikingly absent in scalp hair follicle melanocytes. CONCLUSIONS: The eyelash is structurally very close to curly hair but some biological processes related to follicle cycle and pigmentation differ markedly. PMID- 19804591 TI - Effectiveness of skin protection measures in prevention of occupational hand eczema: results of a prospective randomized controlled trial over a follow-up period of 1 year. AB - BACKGROUND: We recently found a very low adherence to a generally recommended skin protection regimen in a sample of 1355 metalworkers. OBJECTIVES: The present study assessed the effectiveness of skin protection as presently recommended, especially the differential contribution of skin care and skin protection, to the prevention of occupational hand eczema. Methods Of 1355 metalworkers screened, 1020 male volunteers, all fit for work, were recruited for a prospective intervention study with four arms (skin care, skin protection, both combined, and control group, i.e. no recommendation). The study was performed from winter 2006/2007 to spring 2008, following each subject for up for 12 months. Both hands were examined using a quantitative skin score, and a standardized personal interview was performed three times. The change of the objective skin score from baseline to 12 months was used as primary outcome measure. RESULTS: After 12 months 800 subjects were included (78.4% of those recruited). The compliance to follow the randomized measure depended on the recommended measure and ranged from 73.7% to 88.7%. While in the control group a significant deterioration was found, the largest and significant improvement was noted in the group following the generally recommended skin protection programme (skin care + skin protection) followed by skin protection alone as second best. CONCLUSIONS: The generally recommended skin protection regimen seems to provide effective prevention of occupational skin disease. Therefore, the compliance to follow the skin protection regimen, especially the use of skin protection, should be enhanced. PMID- 19804592 TI - A study of matrix metalloproteinase expression and activity in atopic dermatitis using a novel skin wash sampling assay for functional biomarker analysis. AB - BACKGROUND: Atopic dermatitis (AD) is a chronic inflammatory skin condition that is characterized by a defective skin barrier. Despite the well-recognized role of proteases in skin barrier maintenance, relatively little is known of the contribution made by matrix metalloproteinases (MMPs) to the inflammatory process in AD. OBJECTIVES: To test a simple, novel ex vivo bioassay technique in an analysis of the MMPs present in wash samples taken from the skin surface of patients with AD. METHODS: Saline wash samples were collected from eczematous and unaffected areas of the skin of patients with AD and from the skin of normal controls. Wash samples were analysed for their MMP content using a functional peptide cleavage assay, gelatin zymography and an antibody array. RESULTS: Using a functional substrate cleavage assay, skin wash samples from AD lesions were shown to contain 10- to 24-fold more MMP activity than those from normal control skin (P < 0.02) and fivefold more than those from unaffected AD skin (P < 0.05); this activity was inhibited by a broad-spectrum MMP inhibitor Ro 31-9790. Gelatin zymography and antibody array analysis revealed substantial levels of MMP-8 (neutrophil collagenase) and MMP-9 (92-kDa gelatinase) in AD skin wash samples as well as lower levels of MMP-10 (stromelysin 2) and tissue inhibitor of metalloproteinases (TIMP)-1 and TIMP-2; low levels of MMP-1 (fibroblast collagenase), MMP-3 (stromelysin 1) and TIMP-4 were also detected. CONCLUSIONS: A simple skin wash technique suitable for the quantitative and functional analysis of biomolecules in AD is described. Using this method we show that MMPs, and in particular MMP-8 and MMP-9, represent an important potential component of the pathology of AD. The method is expected to prove useful in advancing our understanding of AD and in identifying biomarkers for the evaluation of new therapies. PMID- 19804593 TI - Long-term follow-up of photodynamic therapy with a self-adhesive 5 aminolaevulinic acid patch: 12 months data. AB - BACKGROUND: Photodynamic therapy with a self-adhesive 5-aminolaevulinic acid (5 ALA) patch shows high efficacy rates in the treatment of mild to moderate actinic keratosis (AK) in short term trials. OBJECTIVES: The purpose of the trial was to follow up patients after successful 5-ALA patch-PDT at 3 month intervals over a total period of 12 months. Patients who had received placebo-PDT or cryosurgery served for comparison. PATIENTS/METHODS: Three months after therapy, 360 patients from two separate randomized parallel group phase III studies (one superiority trial vs. placebo-PDT, one noninferiority trial vs. cryosurgery) were suitable for the follow-up study. Patients had to show at least one successfully treated AK lesion after initial therapy. A total of 316 patients completed the follow-up. RESULTS: Twelve months after a single treatment, 5-ALA patch-PDT still proved superior to placebo-PDT and cryosurgery (P < 0.001 for all tests). On a lesion basis, efficacy rates were 63% and 79% for PDT, 63% for cryosurgery and 9% and 25% for placebo-PDT. Recurrence rates of patch-PDT proved superior to those of cryosurgery (per protocol set: P = 0.011, full analysis set: P = 0.049). While 31% of cryosurgery lesions were still hypopigmented after 1 year, the 5-ALA patch PDT groups showed hypopigmentation in 0% (superiority trial) and 3% (noninferiority trial) of the treated lesions. CONCLUSION: Twelve months after a single 5-ALA patch-PDT the majority of lesions were still cleared with an excellent cosmetic outcome. 5-ALA patch-PDT proved to be superior to cryosurgery in the noninferiority study setting. PMID- 19804594 TI - Topical nicotinamide modulates cellular energy metabolism and provides broad spectrum protection against ultraviolet radiation-induced immunosuppression in humans. AB - BACKGROUND: Ultraviolet (UV) radiation can profoundly suppress the cutaneous immune system, thus enhancing carcinogenesis. Agents that prevent UV-induced immunosuppression may thus reduce skin cancer. Nicotinamide (vitamin B3) prevents UV-induced immunosuppression and carcinogenesis in mice, and solar-simulated (ss) UV-induced immunosuppression in humans. Its effectiveness against different UV wavebands and mechanism of action is as yet unknown. OBJECTIVES: To determine the effects and mechanisms of topical nicotinamide on UV-induced suppression of delayed type hypersensitivity (DTH) responses in humans. METHODS: Healthy Mantoux positive volunteers in four randomised, double-blinded studies were irradiated with solar-simulated (ss)UV (UVB + UVA) or narrowband UVB (300 nm) or UVA (385 nm). Topical nicotinamide (0.2% or 5%) or its vehicle were applied immediately after each irradiation. Mantoux testing was performed at irradiated sites and adjacent unirradiated control sites 48 h after the first irradiation and measured 72 h later. Immunosuppression was calculated as the difference in Mantoux-induced erythema and induration at test sites compared to control sites. Human keratinocyte cell cultures, with and without ssUV and nicotinamide, were used for quantitative real-time reverse transcriptase-polymerase chain reaction assessment of TP53 and enzymes that regulate oxidative phosphorylation. RESULTS: Nicotinamide cooperated with ssUV to increase enzymes involved in cellular energy metabolism and p53, and significantly protected against immunosuppression caused by UVB, longwave UVA and single and repeated ssUV exposures. CONCLUSIONS: Longwave UVA, which is poorly filtered by most sunscreens, was highly immune suppressive even at doses equivalent to 20 min of sun exposure. Nicotinamide, which protected against both UVB and UVA, is a promising agent for skin cancer prevention. PMID- 19804595 TI - Management and outcome of metastatic melanoma during pregnancy. AB - BACKGROUND: Although metastatic melanoma occurrence during pregnancy challenges the physician in several ways, only a few studies have been published. OBJECTIVES: Our aim was to investigate therapeutic management together with maternal and fetal outcomes in pregnant women with advanced melanoma. METHODS: A French national retrospective study was conducted in 34 departments of Dermatology or Oncology. All patients with American Joint Committee on Cancer (AJCC) stage III/IV melanoma diagnosed during pregnancy were included. Data regarding melanoma history, pregnancy, treatment, delivery, maternal and infant outcomes were collected. RESULTS: Twenty-two women were included: 10 AJCC stage III and 12 stage IV. Abortion was performed in three patients. Therapeutic abstention during pregnancy was observed in three cases, 14 patients underwent surgery, four patients received chemotherapy and one patient was treated with brain radiotherapy alone. The median gestational age was 36 weeks amenorrhoea. Neither neonatal metastases nor deformities were observed. Placenta metastases were found in one case. Among 18 newborns, 17 are currently alive (median follow up, 17 months); one died of sudden infant death. The 2-year maternal survival rates were 56% (stage III) and 17% (stage IV). CONCLUSIONS: Faced with metastatic melanoma, a majority of women chose to continue with pregnancy, giving birth, based on our samples, to healthy, frequently premature infants. Except during the first trimester of pregnancy, conventional melanoma treatment was applied. No serious side effect was reported, except one case of miscarriage after surgery. Mortality rates do not suggest a worsened prognosis due to pregnancy but larger prospective controlled studies are necessary to assess this specific point. PMID- 19804596 TI - Opinion on moderate/low cancer genetic risk markers in medical practice including comment on the article Genetic contribution to all cancers: the first demonstration using the model of breast cancers from Poland stratified by age at diagnosis and tumour pathology by Lubinski et al., Breast Cancer Res Treat 2008 Apr 15. PMID- 19804597 TI - Comment on the article Genetic contribution to all cancers: the first demonstration using the model of breast cancers from Poland stratified by age at diagnosis and tumour pathology by Lubinski et al., Breast Cancer Res Treat 2008 Apr 15. PMID- 19804598 TI - Other short opinions/comments on moderate/low cancer genetic risk markers in medical practice and the article Genetic contribution to all cancers: the first demonstration using the model of breast cancers from Poland stratified by age at diagnosis and tumour pathology by Lubinski et al., Breast Cancer Res Treat 2008 Apr 15. PMID- 19804599 TI - Principles of genetic predisposition to malignancies. PMID- 19804600 TI - DNA and RNA analyses in detection of genetic predisposition to cancer. PMID- 19804601 TI - BRCA1 testing. PMID- 19804602 TI - MSH2 and MLH1 testing. PMID- 19804603 TI - DNA testing for variants conferring low or moderate increase in the risk of cancer. PMID- 19804604 TI - Hereditary breast and ovarian cancer. PMID- 19804605 TI - Lynch syndrome (HNPCC). PMID- 19804606 TI - MSH6 syndrome. PMID- 19804607 TI - ChIP'ing the mammalian genome: technical advances and insights into functional elements. AB - Characterization of the functional components in mammalian genomes depends on our ability to completely elucidate the genetic and epigenetic regulatory networks of chromatin states and nuclear architecture. Such endeavors demand the availability of robust and effective approaches to characterizing protein-DNA associations in their native chromatin environments. Consider able progress has been made through the applica tion of chromatin immunoprecipitation (ChIP) to study chromatin biology in cells. Coupled with genome-wide analyses, ChIP-based assays enable us to take a global, unbiased and comprehensive view of transcriptional control, epigenetic regulation and chromatin structures, with high precision and versatility. The integrated knowledge derived from these studies is used to decipher gene regulatory networks and define genome organization. In this review, we discuss this powerful approach and its current advances. We also explore the possible future developments of ChIP-based approaches to interrogating long-range chromatin interactions and their impact on the mechanisms regulating gene expression. PMID- 19804608 TI - Physical forces in myelination and repair: a question of balance? AB - A recent report in BMC Cell Biology examines how the balance of extracellular forces and intracellular contractions regulate the shape changes required for oligodendrocyte myelination. A failure of remyelination such as seen in multiple sclerosis could be caused by loss of this balance. PMID- 19804609 TI - Nursing some sense out of Myc. AB - Data recently published in BMC Biology provide insights into the normal physiological function of c-myc in the development and regeneration of the mammary gland and indicate a key role in epithelial cell proliferation, elaboration of ductal alveoli, and the biosynthetic capacity and milk production of the mature organ. PMID- 19804610 TI - Network strategies to understand the aging process and help age-related drug design. AB - Recent studies have demonstrated that network approaches are highly appropriate tools for understanding the extreme complexity of the aging process. Moreover, the generality of the network concept helps to define and study the aging of technological and social networks and ecosystems, which may generate novel concepts for curing age-related diseases. The current review focuses on the role of protein-protein interaction networks (inter-actomes) in aging. Hubs and inter modular elements of both interactomes and signaling networks are key regulators of the aging process. Aging induces an increase in the permeability of several cellular compartments, such as the cell nucleus, introducing gross changes in the representation of network structures. The large overlap between aging genes and genes of age-related major diseases makes drugs that aid healthy aging promising candidates for the prevention and treatment of age-related diseases, such as cancer, atherosclerosis, diabetes and neurodegenerative disorders. We also discuss a number of possible research options to further explore the potential of the network concept in this important field, and show that multi-target drugs (representing 'magic-buckshots' instead of the traditional 'magic bullets') may become an especially useful class of age-related drugs in the future. PMID- 19804611 TI - Human population structure, genome autozygosity and human health. AB - A major transition in human population structure is currently under way, moving from a historical metapopulation, comprising small and mainly rural endogamous communities, to large and increasingly panmictic urban populations. This process is predicted to increase outbreeding, and preliminary data from genomic surveys have helped to quantify the potential magnitude of the effects. Population genetic trends of this nature should result in a reduced burden of recessive disorders, and have a favourable impact on complex diseases influenced by partially recessive genetic variants of smaller effect. The overall outcome is expected to be beneficial for a range of traits associated with human health and disease that show dominance variance. PMID- 19804612 TI - Gene patents and personalized medicine - what lies ahead? AB - Gene patents have generally not impeded biomedical research, but some problems that arise in genetic diagnostics can be attributed to exclusively licensed gene patents. Gene patents for therapeutics have often been litigated but have received surprisingly little public outcry. In stark contrast, genetic diagnostics have been highly controversial but rarely litigated: no case has gone to trial and there is little case law to guide policy. Most recently the Secretary's Advisory Committee for Genetics Health and Society (SACGHS) released a draft report examining how patenting and licensing affect access to clinical genetic testing in the US. The SACGHS reported that patents neither greatly hindered nor facilitated patient access to genetic testing; both the harms and the benefits of patents on genetic diagnostics have been exaggerated. Problems do occur when patents are exclusively licensed to a single provider and no alternative is available. Courts have been changing the thresholds for what can be patented, and how strongly patents can be enforced. Technologies for sequencing, genotyping and gene expression profiling promise to guide clinical decisions in managing common chronic diseases and infectious diseases, and will likely be an integral part of personalized medicine. Developing such genomic tests may require mapping a complex intellectual property landscape and cutting through thickets of patented DNA sequences and related methods. Our preliminary studies have found patent claims that, if strictly enforced, might block the use of multi-gene tests or full-genome sequence data. Yet new technologies promise to reduce the costs of complete genomic sequencing to prices that are comparable to current genetic tests for a single condition. Courts, companies, and policy makers seem unlikely to allow intellectual property to obstruct such technological advance, but prudent policy will depend on careful analysis and foresight. The SACGHS report signals that the US government is paying attention, and increases the odds that policy will foster socially beneficial uses of genetic testing while preserving intellectual property incentives and mitigating the problems that arise from legal monopolies. PMID- 19804613 TI - Development of 'synthetic lethal' strategies to target BRCA1-deficient breast cancer. AB - Recent clinical trials demonstrating the efficacy of poly(ADP-ribose) polymerase (PARP) inhibitors for the treatment of BRCA1-deficient breast cancer have provided support for the 'synthetic lethal' concept of targeted cancer therapeutics. A new study provides further preclinical validation of this concept by demonstrating that BRCA1-deficient mouse mammary tumor cells are selectively sensitive to an inhibitor of the polycomb gene EZH2. The development of polycomb gene inhibitors may provide a novel approach to selectively exploit the molecular alterations in BRCA1-deficient breast tumors. PMID- 19804614 TI - Use of methotrexate therapy is not associated with decreased prevalence of metabolic syndrome. PMID- 19804616 TI - Sodium and brain injury: do we know what we are doing? AB - There is mounting evidence, including the recent report by Maggiore and colleagues, of an association between hypernatremia and mortality in patients with traumatic brain injury. This mandates a re-evaluation of routine administration of agents such as hypertonic saline for the management of intracranial hypertension in those with traumatic brain injury. PMID- 19804617 TI - Thoracic epidural anesthesia in sepsis--is it harmful or protective? AB - Research interest in epidural anesthesia during sepsis has grown over the past years and studies have tried to determine its mechanisms, which should, theoretically, protect organs and reduce morbidity and mortality. However, different experimental approaches in different animal models have provided conflicting results over whether epidural anesthesia has protective or harmful effects and whether these alter depending on the phase of sepsis, the spread of epidural anesthesia or additional supportive therapies. In the future, more standardized research is necessary to integrate the results of all studies, which have been published. PMID- 19804618 TI - Androgens in rheumatoid arthritis: when are they effectors? AB - Neither hormone receptor genes nor plasma androgens seem significantly altered in female subjects before they became affected by rheumatoid arthritis (RA) and, therefore, do not seem to play a role as risk factors for its development. However, serum testosterone levels are inversely correlated with RA activity and dehydro-epiandrosterone sulfate (DHEAS) plasma levels are inversely correlated with both disease duration and clinical severity in patients already affected by active RA. In particular, gonadal and adrenal androgens (that is, testosterone and DHEAS) are significantly decreased in inflamed synovial tissue/fluids during active disease as a consequence of the inflammatory reaction, which supports a pro-inflammatory milieu in RA joints. Recently, male gender has been found to be a major predictor of remission in early RA. PMID- 19804620 TI - Chemical and environmental vector control as a contribution to the elimination of visceral leishmaniasis on the Indian subcontinent: cluster randomized controlled trials in Bangladesh, India and Nepal. AB - BACKGROUND: Bangladesh, India and Nepal are working towards the elimination of visceral leishmaniasis (VL) by 2015. In 2005 the World Health Organization/Training in Tropical Diseases launched an implementation research programme to support integrated vector management for the elimination of VL from Bangladesh, India and Nepal. The programme is conducted in different phases, from proof-of-concept to scaling up intervention. This study was designed in order to evaluate the efficacy of the three different interventions for VL vector management: indoor residual spraying (IRS); long-lasting insecticide treated nets (LLIN); and environmental modification (EVM) through plastering of walls with lime or mud. METHODS: Using a cluster randomized controlled trial we compared three vector control interventions with a control arm in 96 clusters (hamlets or neighbourhoods) in each of the 4 study sites: Bangladesh (one), India (one) and Nepal (two). In each site four villages with high reported VL incidences were included. In each village six clusters and in each cluster five households were randomly selected for sand fly collection on two consecutive nights. Control and intervention clusters were matched with average pre-intervention vector densities.In each site six clusters were randomly assigned to each of the following interventions: indoor residual spraying (IRS); long-lasting insecticide treated nets (LLIN); environmental management (EVM) or control. All the houses (50-100) in each intervention cluster underwent the intervention measures. A reduction of intra-domestic sand fly densities measured in the study households by overnight US Centres for Disease Prevention and Control light trap captures (that is the number of sand flies per trap per night) was the main outcome measure. RESULTS: IRS, and to a lesser extent EVM and LLINs, significantly reduced sand fly densities for at least 5 months in the study households irrespective of type of walls or whether or not people shared their house with cattle. IRS was effective in all sites but LLINs were only effective in Bangladesh and India. Mud plastering did not reduce sand fly density (Bangladesh study); lime plastering in India and one Nepali site, resulted in a significant reduction of sand fly density but not in the second Nepali site. CONCLUSION: Sand fly control can contribute to the regional VL elimination programme; IRS should be strengthened in India and Nepal but in Bangladesh, where vector control has largely been abandoned during the last decades, the insecticide treatment of existing bed nets (coverage above 90% in VL endemic districts) could bring about an immediate reduction of vector populations; operational research to inform policy makers about the efficacious options for VL vector control and programme performance should be strengthened in the three countries. PMID- 19804619 TI - Osteoarthritis associated with estrogen deficiency. AB - Osteoarthritis (OA) affects all articular tissues and finally leads to joint failure. Although articular tissues have long been considered unresponsive to estrogens or their deficiency, there is now increasing evidence that estrogens influence the activity of joint tissues through complex molecular pathways that act at multiple levels. Indeed, we are only just beginning to understand the effects of estrogen deficiency on articular tissues during OA development and progression, as well as on the association between OA and osteoporosis. Estrogen replacement therapy and current selective estrogen receptor modulators have mixed effectiveness in preserving and/or restoring joint tissue in OA. Thus, a better understanding of how estrogen acts on joints and other tissues in OA will aid the development of specific and safe estrogen ligands as novel therapeutic agents targeting the OA joint as a whole organ. PMID- 19804621 TI - Recombination and insertion events involving the botulinum neurotoxin complex genes in Clostridium botulinum types A, B, E and F and Clostridium butyricum type E strains. AB - BACKGROUND: Clostridium botulinum is a taxonomic designation for at least four diverse species that are defined by the expression of one (monovalent) or two (bivalent) of seven different C. botulinum neurotoxins (BoNTs, A-G). The four species have been classified as C. botulinum Groups I-IV. The presence of bont genes in strains representing the different Groups is probably the result of horizontal transfer of the toxin operons between the species. RESULTS: Chromosome and plasmid sequences of several C. botulinum strains representing A, B, E and F serotypes and a C. butyricum type E strain were compared to examine their genomic organization, or synteny, and the location of the botulinum toxin complex genes. These comparisons identified synteny among proteolytic (Group I) strains or nonproteolytic (Group II) strains but not between the two Groups. The bont complex genes within the strains examined were not randomly located but found within three regions of the chromosome or in two specific sites within plasmids. A comparison of sequences from a Bf strain revealed homology to the plasmid pCLJ with similar locations for the bont/bv b genes but with the bont/a4 gene replaced by the bont/f gene. An analysis of the toxin cluster genes showed that many recombination events have occurred, including several events within the ntnh gene. One such recombination event resulted in the integration of the bont/a1 gene into the serotype toxin B ha cluster, resulting in a successful lineage commonly associated with food borne botulism outbreaks. In C. botulinum type E and C. butyricum type E strains the location of the bont/e gene cluster appears to be the result of insertion events that split a rarA, recombination-associated gene, independently at the same location in both species. CONCLUSION: The analysis of the genomic sequences representing different strains reveals the presence of insertion sequence (IS) elements and other transposon-associated proteins such as recombinases that could facilitate the horizontal transfer of the bonts; these events, in addition to recombination among the toxin complex genes, have led to the lineages observed today within the neurotoxin-producing clostridia. PMID- 19804622 TI - The consequences of delaying insulin initiation in UK type 2 diabetes patients failing oral hyperglycaemic agents: a modelling study. AB - BACKGROUND: Recent data have shown that type 2 diabetes patients in the UK delay initiating insulin on average for over 11 years after first being prescribed an oral medication. Using a published computer simulation model of diabetes we used UK-specific data to estimate the clinical consequences of immediately initiating insulin versus delaying initiation for periods in line with published estimates. METHODS: In the base case scenario simulated patients, with characteristics based on published UK data, were modelled as either initiating insulin immediately or delaying for 8 years. Clinical outcomes in terms of both life expectancy and quality-adjusted life expectancy and also diabetes-related complications (cumulative incidence and time to onset) were projected over a 35 year time horizon. Treatment effects associated with insulin use were taken from published studies and sensitivity analyses were performed around time to initiation of insulin, insulin efficacies and hypoglycaemia utilities. RESULTS: For patients immediately initiating insulin there were increases in (undiscounted) life expectancy of 0.61 years and quality-adjusted life expectancy of 0.34 quality adjusted life years versus delaying initiation for 8 years. There were also substantial reductions in cumulative incidence and time to onset of all diabetes related complications with immediate versus delayed insulin initiation. Sensitivity analyses showed that a reduced delay in insulin initiation or change in insulin efficacy still demonstrated clinical benefits for immediate versus delayed initiation. CONCLUSION: UK type 2 diabetes patients are at increased risk of a large number of diabetes-related complications due to an unnecessary delay in insulin initiation. Despite clear guidelines recommending tight glycaemic control this failure to begin insulin therapy promptly is likely to result in needlessly reduced life expectancy and compromised quality of life. PMID- 19804623 TI - Age at menarche and the menstrual pattern of secondary school adolescents in northwest Ethiopia. AB - BACKGROUND: Population studies on normal and dysfunctional characteristics of menstrual cycles are scarce in Ethiopia. In addition variability in menarcheal age and menstrual characteristics are common. Knowledge on this variability is necessary for patient education and to guide clinical evaluation. METHODS: A cross sectional study was conducted in two small towns called Dabat and Kola Diba, northwest Ethiopia between April and May 2007. Systematic sampling method was used to select 622 school girls from two secondary schools. A pretested questionnaire prepared in Amharic was used to gather data. Selected girls cooperated in answering the questionnaire in their classrooms under the supervision of the research team. Only 612 of the adolescent females were included in the final analysis, of which 305 were from Koladiba High School and 307 from Dabat. RESULTS: The age of the study subjects ranges between 14 and 19 with a mean (standard deviation) of 16.9 +/- 1 years. About 92.2% had attained menarche by the time the survey was conducted. The probit analysis of the status quo data yielded a median (CI) age at menarche of 14.8 (13.9-15.3) years. The average age at menarche by recall method was 15.8 +/- 1 years. The mean age at menarche was 0.3 years younger for urban females compared with rural ones (p < 0.001). A cycle length between 21 and 35 days was observed in 70.3% of the girls. The mean duration of flow was 4 +/- 1.3 days with a range of 2-7 days. The menstrual cycles were irregular in 42.8% of the subjects. The overall prevalence of dysmenorrhoea was 72% among these subjects. Premenstrual symptoms were present in 435 of the females (75.4%). The leading sources of menarcheal information to the adolescents were mothers (39.7%), followed by their friends (26.6%) and teachers (21.8%). CONCLUSION: In this study age of menarche was found to be delayed which is even higher than the findings indicated similar studies conducted in Ethiopia and other African countries. A significant number of students complain of abnormal menstrual cycle, dysmenorrhoea and premenstrual symptoms which call for appropriate counselling and management. PMID- 19804624 TI - A case study evaluation of implementation of a care pathway to support normal birth in one English birth centre: anticipated benefits and unintended consequences. AB - BACKGROUND: The policy drive for the UK National Health Service (NHS) has focused on the need for high quality services informed by evidence of best practice. The introduction of care pathways and protocols to standardise care and support implementation of evidence into practice has taken place across the NHS with limited evaluation of their impact. A multi-site case study evaluation was undertaken to assess the impact of use of care pathways and protocols on clinicians, service users and service delivery. One of the five sites was a midwifery-led Birth Centre, where an adapted version of the All Wales Clinical Pathway for Normal Birth had been implemented. METHODS: The overarching framework was realistic evaluation. A case study design enabled the capture of data on use of the pathway in the clinical setting, use of multiple methods of data collection and opportunity to study and understand the experiences of clinicians and service users whose care was informed by the pathway. Women attending the Birth Centre were recruited at their 36 week antenatal visit. Episodes of care during labour were observed, following which the woman and the midwife who cared for her were interviewed about use of the pathway. Interviews were also held with other key stakeholders from the study site. Qualitative data were content analysed. RESULTS: Observations were undertaken of four women during labour. Eighteen interviews were conducted with clinicians and women, including the women whose care was observed and the midwives who cared for them, senior midwifery managers and obstetricians. The implementation of the pathway resulted in a number of anticipated benefits, including increased midwifery confidence in skills to support normal birth and promotion of team working. There were also unintended consequences, including concerns about a lack of documentation of labour care and negative impact on working relationships with obstetric and other midwifery colleagues. Women were unaware their care was informed by a care pathway. CONCLUSION: Care pathways are complex interventions which generate a number of consequences for practice. Those considering introduction of pathways need to ensure all relevant stakeholders are engaged with this and develop robust evaluation strategies to accompany implementation. PMID- 19804625 TI - Chemokine receptor expression and functional effects of chemokines on B cells: implication in the pathogenesis of rheumatoid arthritis. AB - INTRODUCTION: Accumulation of B cells in the rheumatoid arthritis (RA) synovium has been reported, and it has been thought that these cells might contribute to the pathogenesis of RA by antigen presentation, autoantibody production, and/or inflammatory cytokine production. Chemokines could enhance the accumulation of B cells in the synovium. The aims of this study were to determine chemokine receptor expression by B cells both in the peripheral blood of normal donors and subjects with RA, and at the inflammatory site in RA, and the effects of chemokines on B cell activation. METHODS: Cell surface molecule expression was analyzed by flow cytometry. Cellular migration was assessed using chemotaxis chambers. Cellular proliferation was examined by 3H-thymidine incorporation. Tumor necrosis factor (TNF) production was assayed by enzyme-linked immunosorbent assay. RESULTS: Significant numbers of peripheral blood B cells of healthy donors and subjects with RA expressed CC chemokine receptor (CCR)5 and CXCR3, and most B cells expressed CCR6, CCR7, CXCR4 and CXCR5. CCR5 expression was more frequent on CD27+ than CD27- peripheral blood B cells of healthy donors and RA. Synovial B cells more frequently expressed CCR5, but less often expressed CCR6, CCR7 and CXCR5 compared to peripheral blood in RA. Further functional analyses were performed on peripheral blood B cells from healthy donors. Migration of peripheral blood B cells, especially CD27+ B cells, was enhanced by CC chemokine ligand (CCL)20, CCL19, CCL21 and CXCL12. All four chemokines alone induced B cell proliferation; with CCL21 being the most effective. CCL21 also enhanced the proliferation of anti-immunoglobulin (Ig)M-stimulated B cells and blockade of CCR7 inhibited this effect. CCL20, CCL21 and CXCL12 enhanced TNF production by anti-IgM mAb-stimulated B cells. Finally, stimulation with CXCL12, but not CCL20, CCL19 and CCL21, enhanced inducible costimulator-ligand (ICOSL) expression by peripheral blood B cells of healthy donors and RA, but did not increase B cell activating factor receptor or transmembrane activator and CAML-interactor. CONCLUSIONS: The data suggest that CCR5, CCR6, CCR7, CXCR3, CXCR4 and CXCR5 may be important for the B cell migration into the synovium of RA patients, and also their local proliferation, cytokine production and ICOSL expression in the synovium. PMID- 19804626 TI - Treatment of candidemia and invasive candidiasis in the intensive care unit: post hoc analysis of a randomized, controlled trial comparing micafungin and liposomal amphotericin B. AB - INTRODUCTION: Invasive candidiasis and candidemia are life-threatening nosocomial infections in intensive care patients. METHODS: A post hoc analysis of a phase 3 trial assessing micafungin (100 mg/day for subjects > 40 kg; 2 mg/kg/day for subjects 25) in 22% of young healthy subjects. Increased cholesterol values (>5.0 mmol/L) were found in 23% of subjects, LDL-C (>3.0 mmol/L) in 23%, triglycerides (>1.7 mmol/L) in 11% of subjects. We found statistically significant differences in subjects' weight (p = 0.015), BMI (p = 0.023), and waist-hip ratio (WHR) (p = 0.015) in regard to their diet type; subjects with Mediterranean diet had the lowest values compared to those on continental and mixed diet. Significant associations were found for: LPL genetic polymorphic variant and abdominal obesity (p = 0.013), APO epsilon4 allele and hypercholesterolemia (p = 0.003), and ESR1-TA long allele and hypercholesterolemia (p = 0.011). BACKGROUND: Human obesity is a multifactorial syndrome influenced also by genetic factors. Among gene variants found to be involved in body weight regulation and development of obesity, particular attention has been paid to polymorphisms in genes associated with obesity-related metabolic disorders. We explored the association of genetic polymorphisms of: estrogen receptor alpha (ESR1-TA repeats); interleukin-6 (IL-6 G-174C); apolipoprotein E (APO epsilon2, epsilon3, epsilon4); lipoprotein lipase Pvu II (LPL P+/-), with clinical variables: gender, age, body mass index (BMI), diet type and biological variables: triglycerides, cholesterol, HDL-C, LDL-C, CRP, homocysteine, urate, and glucose in 105 healthy young subjects (20-35 yrs) of Croatian origin. METHODS: Genotyping of IL-6, LPL was performed by PCR-RFLP, of APOE by real-time PCR, and of ESR1 by PCR and capillary electrophoresis. Association analyses were performed of alleles and genotypes with biological variables. CONCLUSION: ESR-1, LPL, and APO E genetic polymorphic variants could represent predictive genetic risk markers for obesity-related metabolic disorders in young healthy subjects. Mediterranean type of diet is also an important protective factor against abdominal obesity. PMID- 19804634 TI - Effects of positive end-expiratory pressure on respiratory function and hemodynamics in patients with acute respiratory failure with and without intra abdominal hypertension: a pilot study. AB - INTRODUCTION: To investigate the effects of positive end-expiratory pressure (PEEP) on respiratory function and hemodynamics in patients with acute lung injury (ALI) or acute respiratory distress syndrome (ARDS) with normal intra abdominal pressure (IAP < 12 mmHg) and with intra-abdominal hypertension (IAH, defined as IAP >or= 12 mmHg) during lung protective ventilation and a decremental PEEP, a prospective, observational clinical pilot study was performed. METHODS: Twenty patients with ALI/ARDS with normal IAP or IAH treated in the surgical intensive care unit in a university hospital were studied. The mean IAP in patients with IAH and normal IAP was 16 +/- 3 mmHg and 8 +/- 3 mmHg, respectively (P < 0.001). At different PEEP levels (5, 10, 15, 20 cmH2O) we measured respiratory mechanics, partitioned into its lung and chest wall components, alveolar recruitment, gas-exchange, hemodynamics, extravascular lung water index (EVLWI) and intrathoracic blood volume index (ITBVI). RESULTS: We found that ALI/ARDS patients with IAH, as compared to those with normal IAP, were characterized by: a) no differences in gas-exchange, respiratory mechanics, partitioned into its lung and chest wall components, as well as hemodynamics and EVLWI/ITBVI; b) decreased elastance of the respiratory system and the lung, but no differences in alveolar recruitment and oxygenation or hemodynamics, when PEEP was increased at 10 and 15cmH2O; c) at higher levels of PEEP, EVLWI was lower in ALI/ARDS patients with IAH as compared with those with normal IAP. CONCLUSIONS: IAH, within the limits of IAP measured in the present study, does not affect interpretation of respiratory mechanics, alveolar recruitment and hemodynamics. PMID- 19804635 TI - Inhibition of extracellular matrix assembly induces the expression of osteogenic markers in skeletal muscle cells by a BMP-2 independent mechanism. AB - BACKGROUND: The conversion of one cell type into another has been suggested to be, at the molecular level, the consequence of change(s) in the expression level of key developmental genes. Myoblasts have the ability to differentiate either to skeletal muscle or osteogenic lineage depending of external stimuli. Extracellular matrix (ECM) has been shown to be essential for skeletal muscle differentiation, through its direct interaction with myoblasts' cell receptors. We attempt to address if ECM also plays a role in the osteogenic differentiation of skeletal muscle cells. RESULTS: Inhibition of proteoglycan sulfation by sodium chlorate in myoblast cultures strongly affects ECM synthesis and deposition and induces the expression of the osteogenic lineage markers alkaline phosphatase (ALP) and osteocalcin in mononuclear cells. Induction of ALP by sodium chlorate does not affect the expression of specific muscle determination transcription factors, such as MyoD and Myf-5, in the same cells. The osteogenic transcription factor Cbfa-1 expression is also unaffected. Induction of ALP is not inhibited by a soluble form of BMP receptor IA. This suggests that the deviation of the myogenic pathway of C2C12 myoblasts into the osteogenic lineage by inhibitors of proteoglycan sulfation is BMP-2 independent. The increase of osteogenic markers expression can be totally prevented by an exogenous ECM. Interestingly, a similar BMP-2-independent ALP activity induction can be observed in myoblasts cultured on an ECM previously synthesized by BMP-2 treated myoblasts. Under in vivo conditions of increased ECM turn-over and deposition, as in the mdx dystrophic muscle and during skeletal muscle regeneration, an induction and relocalization of ALP is observed in a subpopulation of skeletal muscle fibers, whereas in normal skeletal muscle, ALP expression is restricted to blood vessels and some endomysial mononuclear cells. CONCLUSION: These results suggest that signals arising from the ECM induce the expression of osteogenic markers in muscle cells by a mechanism independent of BMP-2 and without affecting the expression of key muscle or osteogenic determination genes. An induction and relocalization of ALP is also observed in mdx and regenerating skeletal muscles, in vivo conditions of increased muscle ECM deposition or turnover. PMID- 19804636 TI - Optimization and comparison of different methods for RNA isolation for cDNA library construction from the reindeer lichen Cladonia rangiferina. AB - BACKGROUND: The reindeer lichen is the product of a mutualistic relationship between a fungus and an algae. Lichen demonstrate a remarkable capacity to tolerate dehydration. This tolerance is driven by a variety of biochemical processes and the accumulation of specific secondary metabolites that may be of relevance to the pharmaceutical, biotechnology and agriculture industries. These protective metabolites hinder in vitro enzymatic reactions required in cDNA synthesis. Along with the low concentrations of RNA present within lichen tissues, the process of creating a cDNA library is technically challenging. FINDINGS: An evaluation of existing commercial and published protocols for RNA extraction from plant or fungal tissues has been performed and experimental conditions have been optimised to balance the need for the highest quality total ribonucleotides and the constraints of budget, time and human resources. CONCLUSION: We present a protocol that balances inexpensive RNA extraction methods with commercial RNA clean-up kits to yield sufficient RNA for cDNA library construction. Evaluation of the protocol and the construction of, and sampling from, a cDNA library is used to demonstrate the suitability of the RNA extraction method for expressed sequence tag production. PMID- 19804637 TI - High adsorption rate is detrimental to bacteriophage fitness in a biofilm-like environment. AB - BACKGROUND: Bacterial biofilm is ubiquitous in nature. However, it is not clear how this crowded habitat would impact the evolution of bacteriophage (phage) life history traits. In this study, we constructed isogenic lambda phage strains that only differed in their adsorption rates, because of the presence/absence of extra side tail fibers or improved tail fiber J, and maker states. The high cell density and viscosity of the biofilm environment was approximated by the standard double-layer agar plate. The phage infection cycle in the biofilm environment was decomposed into three stages: settlement on to the biofilm surface, production of phage progeny inside the biofilm, and emigration of phage progeny out of the current focus of infection. RESULTS: We found that in all cases high adsorption rate is beneficial for phage settlement, but detrimental to phage production (in terms of plaque size and productivity) and emigration out of the current plaque. Overall, the advantage of high adsorption accrued during settlement is more than offset by the disadvantages experienced during the production and emigration stages. The advantage of low adsorption rate was further demonstrated by the rapid emergence of low-adsorption mutant from a high-adsorption phage strain with the side tail fibers. DNA sequencing showed that 19 out of the 21 independent mutant clones have mutations in the stf gene, with the majority of them being single-nucleotide insertion/deletion mutations occurring in regions with homonucleotide runs. CONCLUSION: We conclude that high mutation rate of the stf gene would ensure the existence of side tail fiber polymorphism, thus contributing to rapid adaptation of the phage population between diametrically different habitats of benthic biofilm and planktonic liquid culture. Such adaptability would also help to explain the maintenance of the stf gene in phage lambda's genome. PMID- 19804638 TI - MethMarker: user-friendly design and optimization of gene-specific DNA methylation assays. AB - DNA methylation is a key mechanism of epigenetic regulation that is frequently altered in diseases such as cancer. To confirm the biological or clinical relevance of such changes, gene-specific DNA methylation changes need to be validated in multiple samples. We have developed the MethMarker http://methmarker.mpi-inf.mpg.de/ software to help design robust and cost efficient DNA methylation assays for six widely used methods. Furthermore, MethMarker implements a bioinformatic workflow for transforming disease-specific differentially methylated genomic regions into robust clinical biomarkers. PMID- 19804639 TI - Replication of an empirical approach to delineate the heterogeneity of chronic unexplained fatigue. AB - BACKGROUND: Chronic fatigue syndrome (CFS) is defined by self-reported symptoms. There are no diagnostic signs or laboratory markers, and the pathophysiology remains inchoate. In part, difficulties identifying and replicating biomarkers and elucidating the pathophysiology reflect the heterogeneous nature of the syndromic illness CFS. We conducted this analysis of people from defined metropolitan, urban, and rural populations to replicate our earlier empirical delineation of medically unexplained chronic fatigue and CFS into discrete endophenotypes. Both the earlier and current analyses utilized quantitative measures of functional impairment and symptoms as well as laboratory data. This study and the earlier one enrolled participants from defined populations and measured the internal milieu, which differentiates them from studies of clinic referrals that examine only clinical phenotypes. METHODS: This analysis evaluated 386 women identified in a population-based survey of chronic fatigue and unwellness in metropolitan, urban, and rural populations of the state of Georgia, USA. We used variables previously demonstrated to effectively delineate endophenotypes in an attempt to replicate identification of these endophenotypes. Latent class analyses were used to derive the classes, and these were compared and contrasted to those described in the previous study based in Wichita, Kansas. RESULTS: We identified five classes in the best fit analysis. Participants in Class 1 (25%) were polysymptomatic, with sleep problems and depressed mood. Class 2 (24%) was also polysymptomatic, with insomnia and depression, but participants were also obese with associated metabolic strain. Class 3 (20%) had more selective symptoms but was equally obese with metabolic strain. Class 4 (20%) and Class 5 (11%) consisted of nonfatigued, less symptomatic individuals, Class 4 being older and Class 5 younger. The classes were generally validated by independent variables. People with CFS fell equally into Classes 1 and 2. Similarities to the Wichita findings included the same four main defining variables of obesity, sleep problems, depression, and the multiplicity of symptoms. Four out of five classes were similar across both studies. CONCLUSION: These data support the hypothesis that chronic medically unexplained fatigue is heterogeneous and can be delineated into discrete endophenotypes that can be replicated. The data do not support the current perception that CFS represents a unique homogeneous disease and suggests broader criteria may be more explanatory. This replication suggests that delineation of endophenotypes of CFS and associated ill health may be necessary in order to better understand etiology and provide more patient-focused treatments. PMID- 19804640 TI - Plasma homocysteine in adolescents depends on the interaction between methylenetetrahydrofolate reductase genotype, lipids and folate: a seroepidemiological study. AB - BACKGROUND: Many publications link high homocysteine levels to cardiovascular disease. In Spain there is little information on the prevalence of hyperhomocysteinaemia and associated vitamin factors among the general population, and less still among children. Cardiovascular risk factors in the childhood population may be related to the appearance of cardiovascular disease at adult age. The aim of this study is to establish a definition of hyperhomocysteinaemia in adolescents and to analyze the influence of vitamin and metabolic factors in homocysteine levels in this population group. METHODS: Descriptive, cross-sectional epidemiological study to estimate serum homocysteine, vitamin B12 and folate levels, as well as plasma total, HDL- and LDL- cholesterol in a schoolgoing population aged 13 to 17 years in Madrid, Spain.Spearman correlation analysis was performed to ascertain quantitative comparison, Pearson's chi2 test (frequency < 5, Fisher) was used for comparison of prevalences, Mann-Whitney U and Kruskal-Wallis test were used for comparison of means and Bonferroni correction was used for post-hoc tests. A multivariate logistic regression model was performed in the multivariate analysis. RESULTS: Based on the classic values for definition of hyperhomocysteinaemia in adults, prevalence of hyperhomocysteinaemia in the study population was: 1.26% for 15 mumol/L; and 2.52% for 12 mumol/L.Deficits in HDL cholesterol and serum folate levels yielded adjusted Odds Ratios (OR) for hyperhomocysteinemia of 2.786, 95% CI (1.089-7.126), and 5.140, 95% CI (2.347-11.256) respectively. Mutation of the methylenetetrahydrofolate reductase (MTHFR) C677T genotype also raises the risk of hyperhomocysteinaemia (CC-->CT: OR = 2.362; 95% CI (1.107-5.042) CC-->TT: OR = 6.124, 95% CI (2.301-16.303)) CONCLUSION: A good definition of hyperhomocysteinaemia in adolescents is the 90th percentile, equivalent to 8.23 mumol/L. Risk factors for hyperhomocysteinaemia are cHDL and folate deficiency, and the MTHFR C677T mutant genotype. No significant effect could be assessed for vitamin B12. Coexistence of all three factors increases the risk of suffering from hyperhomocysteinaemia 87-fold. PMID- 19804641 TI - Effects of apple juice on risk factors of lipid profile, inflammation and coagulation, endothelial markers and atherosclerotic lesions in high cholesterolemic rabbits. AB - BACKGROUND: Atherosclerosis which results from gradual deposition of lipids in medium and large arteries is a leading cause of mortality worldwide. The objective of this study was to determine the effect of apple juice on some risk factors of atherosclerosis and on the development of atherosclerosis in rabbits fed a high-cholesterol diet. METHODS: Thirty two male rabbits were randomly divided into four groups: normal diet, high cholesterol diet (%1 cholesterol), 1% cholesterol supplemented with 5 ml apple juice (low dose) and 1% cholesterol supplemented with 10 ml apple juice (high dose) for 2 month. The C-reactive protein (CRP), nitrite, nitrate, fibrinogen, total cholesterol(TC) and factor VII were measured before the experiment and by the end of period. At the end of study, fatty streak formation in right and left coronary arteries were determined using Chekanov method in all groups. RESULTS: Both doses of apple juice significantly were decreased TC, TG, CRP, fibrinogen, factor VII levels, atherosclerotic lesion in right and left coronary arteries and increased nitrite and nitrate compared to cholesterolemic diet. Also using 10 ml apple juice caused significant reduce in LDL-C and increase HDL-C, but 5 ml apple juice did not change these factors. Significant differences were observed between 5 and 10 ml apple juice groups by LDL-C. No significant difference was found between 5 and 10 ml apple juice groups with regard to CRP, nitrite, nitrate, fibrinogen, factor VII, TG, HDL-C and TC concentrations. CONCLUSION: Apple juice can effectively prevent the progress of atherosclerosis. This is likely due to antioxidant and anti-inflammatory effect of apple juice. PMID- 19804642 TI - Molecular allergology approach to allergic diseases in the paediatric age. AB - Identification, characterization, and purification of allergens are essential for the structural and immunologic studies needed to understand how these molecules induce specific IgE antibody production by the human immune system. Advances in molecular biology techniques have led to the production of recombinant allergens having constant properties, allowing detection of specific IgE directed against different molecular components of an allergenic source. Presence of homologous allergens in different sources is the reason for cross reaction. Molecule-based diagnostic tools can lead to better interpretation of poly-sensitizations, observed by ST and in vitro tests using allergenic extracts as they were made before. Some examples IgE sensitization to major genuine allergens and panallergens will be presented. PMID- 19804643 TI - Frequency of medically attended adverse events following tetanus and diphtheria toxoid vaccine in adolescents and young adults: a Vaccine Safety Datalink study. AB - BACKGROUND: Local reactions are the most commonly reported adverse events following tetanus and diphtheria toxoid (Td) vaccine and the risk of local reactions may increase with number of prior Td vaccinations. METHODS: To estimate the risk of medically attended local reactions following Td vaccination in adolescents and young adults we conducted a six-year retrospective cohort study assessing 436,828 Td vaccinations given to persons 9 through 25 years of age in the Vaccine Safety Datalink population from 1999 through 2004. RESULTS: Overall, the estimated risk of a medically attended local reaction was 3.6 events per 10,000 Td vaccinations. The lowest risk (2.8 events per 10,000 vaccinations) was found in the 11 to 15 year old age group. In comparison with that group, the event risks were significantly higher in both the 9 to 10 and 21 to 25 year old age groups. The risk of a local reaction was significantly higher in persons who had received another tetanus and diphtheria toxoid containing vaccine (TDCV) in the previous five years (incidence rate ratio, 2.9; 95% confidence interval, 1.2 to 7.2). Twenty-eight percent of persons with a local reaction to Td vaccine were prescribed antibiotics. CONCLUSION: Medically attended local reactions were uncommon following Td vaccination. The risk of those reactions varied by age and by prior receipt of TDCVs. These findings provide a point of reference for future evaluations of the safety profile of newer vaccines containing tetanus or diphtheria toxoid. PMID- 19804644 TI - High resolution analysis of the human transcriptome: detection of extensive alternative splicing independent of transcriptional activity. AB - BACKGROUND: Commercially available microarrays have been used in many settings to generate expression profiles for a variety of applications, including target selection for disease detection, classification, profiling for pharmacogenomic response to therapeutics, and potential disease staging. However, many commercially available microarray platforms fail to capture transcript diversity produced by alternative splicing, a major mechanism for driving proteomic diversity through transcript heterogeneity. RESULTS: The human Genome-Wide SpliceArray(TM) (GWSA), a novel microarray platform, utilizes an existing probe design concept to monitor such transcript diversity on a genome scale. The human GWSA allows the detection of alternatively spliced events within the human genome through the use of exon body and exon junction probes to provide a direct measure of each transcript, through simple calculations derived from expression data. This report focuses on the performance and validation of the array when measured against standards recently published by the Microarray Quality Control (MAQC) Project. The array was shown to be highly quantitative, and displayed greater than 85% correlation with the HG-U133 Plus 2.0 array at the gene level while providing more extensive coverage of each gene. Almost 60% of splice events among genes demonstrating differential expression of greater than 3 fold also contained extensive splicing alterations. Importantly, almost 10% of splice events within the gene set displaying constant overall expression values had evidence of transcript diversity. Two examples illustrate the types of events identified: LIM domain 7 showed no differential expression at the gene level, but demonstrated deregulation of an exon skip event, while erythrocyte membrane protein band 4.1 like 3 was differentially expressed and also displayed deregulation of a skipped exon isoform. CONCLUSION: Significant changes were detected independent of transcriptional activity, indicating that the controls for transcript generation and transcription are distinct, and require novel tools in order to detect changes in specific transcript quantity. Our results demonstrate that the SpliceArray(TM) design will provide researchers with a robust platform to detect and quantify specific changes not only in overall gene expression, but also at the individual transcript level. PMID- 19804645 TI - Amphioxus (Branchiostoma floridae) has orthologs of vertebrate odorant receptors. AB - BACKGROUND: A common feature of chemosensory systems is the involvement of G protein-coupled receptors (GPCRs) in the detection of environmental stimuli. Several lineages of GPCRs are involved in vertebrate olfaction, including trace amine-associated receptors, type 1 and 2 vomeronasal receptors and odorant receptors (ORs). Gene duplication and gene loss in different vertebrate lineages have lead to an enormous amount of variation in OR gene repertoire among species; some fish have fewer than 100 OR genes, while some mammals possess more than 1000. Fascinating features of the vertebrate olfactory system include allelic exclusion, where each olfactory neuron expresses only a single OR gene, and axonal guidance where neurons expressing the same receptor project axons to common glomerulae. By identifying homologous ORs in vertebrate and in non vertebrate chordates, we hope to expose ancestral features of the chordate olfactory system that will help us to better understand the evolution of the receptors themselves and of the cellular components of the olfactory system. RESULTS: We have identified 50 full-length and 11 partial ORs in Branchiostoma floridae. No ORs were identified in Ciona intestinalis. Phylogenetic analysis places the B. floridae OR genes in a monophyletic clade with the vertebrate ORs. The majority of OR genes in amphioxus are intronless and many are also tandemly arrayed in the genome. By exposing conserved amino acid motifs and testing the ability of those motifs to discriminate between ORs and non-OR GPCRs, we identified three OR-specific amino acid motifs common in cephalochordate, fish and mammalian and ORs. CONCLUSION: Here, we show that amphioxus has orthologs of vertebrate ORs. This conclusion demonstrates that the receptors, and perhaps other components of vertebrate olfaction, evolved at least 550 million years ago. We have also identified highly conserved amino acid motifs that may be important for maintaining receptor conformation or regulating receptor activity. We anticipate that the identification of vertebrate OR orthologs in amphioxus will lead to an improved understanding of OR gene family evolution, OR gene function, and the mechanisms that control cell-specific expression, axonal guidance, signal transduction and signal integration. PMID- 19804646 TI - Global gene expression patterns in the post-pneumonectomy lung of adult mice. AB - BACKGROUND: Adult mice have a remarkable capacity to regenerate functional alveoli following either lung resection or injury that exceeds the regenerative capacity observed in larger adult mammals. The molecular basis for this unique capability in mice is largely unknown. We examined the transcriptomic responses to single lung pneumonectomy in adult mice in order to elucidate prospective molecular signaling mechanisms used in this species during lung regeneration. METHODS: Unilateral left pneumonectomy or sham thoracotomy was performed under general anesthesia (n = 8 mice per group for each of the four time points). Total RNA was isolated from the remaining lung tissue at four time points post-surgery (6 hours, 1 day, 3 days, 7 days) and analyzed using microarray technology. RESULTS: The observed transcriptomic patterns revealed mesenchymal cell signaling, including up-regulation of genes previously associated with activated fibroblasts (Tnfrsf12a, Tnc, Eln, Col3A1), as well as modulation of Igf1-mediated signaling. The data set also revealed early down-regulation of pro-inflammatory cytokine transcripts and up-regulation of genes involved in T cell development/function, but few similarities to transcriptomic patterns observed during embryonic or post-natal lung development. Immunohistochemical analysis suggests that early fibroblast but not myofibroblast proliferation is important during lung regeneration and may explain the preponderance of mesenchymal associated genes that are over-expressed in this model. This again appears to differ from embryonic alveologenesis. CONCLUSION: These data suggest that modulation of mesenchymal cell transcriptome patterns and proliferation of S100A4 positive mesenchymal cells, as well as modulation of pro-inflammatory transcriptome patterns, are important during post-pneumonectomy lung regeneration in adult mice. PMID- 19804647 TI - Low oxygen levels as a trigger for enhancement of respiratory metabolism in Saccharomyces cerevisiae. AB - BACKGROUND: The industrially important yeast Saccharomyces cerevisiae is able to grow both in the presence and absence of oxygen. However, the regulation of its metabolism in conditions of intermediate oxygen availability is not well characterised. We assessed the effect of oxygen provision on the transcriptome and proteome of S. cerevisiae in glucose-limited chemostat cultivations in anaerobic and aerobic conditions, and with three intermediate (0.5, 1.0 and 2.8% oxygen) levels of oxygen in the feed gas. RESULTS: The main differences in the transcriptome were observed in the comparison of fully aerobic, intermediate oxygen and anaerobic conditions, while the transcriptome was generally unchanged in conditions receiving different intermediate levels (0.5, 1.0 or 2.8% O2) of oxygen in the feed gas. Comparison of the transcriptome and proteome data suggested post-transcriptional regulation was important, especially in 0.5% oxygen. In the conditions of intermediate oxygen, the genes encoding enzymes of the respiratory pathway were more highly expressed than in either aerobic or anaerobic conditions. A similar trend was also seen in the proteome and in enzyme activities of the TCA cycle. Further, genes encoding proteins of the mitochondrial translation machinery were present at higher levels in all oxygen limited and anaerobic conditions, compared to fully aerobic conditions. CONCLUSION: Global upregulation of genes encoding components of the respiratory pathway under conditions of intermediate oxygen suggested a regulatory mechanism to control these genes as a response to the need of more efficient energy production. Further, cells grown in three different intermediate oxygen levels were highly similar at the level of transcription, while they differed at the proteome level, suggesting post-transcriptional mechanisms leading to distinct physiological modes of respiro-fermentative metabolism. PMID- 19804648 TI - NADPH oxidase mediates beta-amyloid peptide-induced activation of ERK in hippocampal organotypic cultures. AB - BACKGROUND: Previous studies have shown that beta amyloid (Abeta) peptide triggers the activation of several signal transduction cascades in the hippocampus, including the extracellular signal-regulated kinase (ERK) cascade. In this study we sought to characterize the cellular localization of phosphorylated, active ERK in organotypic hippocampal cultures after acute exposure to either Abeta (1-42) or nicotine. RESULTS: We observed that Abeta and nicotine increased the levels of active ERK in distinct cellular localizations. We also examined whether phospho-ERK was regulated by redox signaling mechanisms and found that increases in active ERK induced by Abeta and nicotine were blocked by inhibitors of NADPH oxidase. CONCLUSION: Our findings indicate that NADPH oxidase-dependent redox signaling is required for Abeta-induced activation of ERK, and suggest a similar mechanism may occur during early stages of Alzheimer's disease. PMID- 19804649 TI - Hepatitis B virus inhibition in mice by lentiviral vector mediated short hairpin RNA. AB - BACKGROUND: Chronic hepatitis B virus (HBV) infection is an important cause of cirrhosis and hepatocellular carcinoma. The major challenges for current therapies are the low efficacy of current drugs and the occurrence of drug resistant HBV mutations. RNA interference (RNAi) of virus-specific genes offers the possibility of developing a new anti-HBV therapy. Recent reports have shown that lentiviral vectors based on HIV-1 are promising gene delivery vehicles due to their ability to integrate transgenes into non-dividing cells. Herein, a lentivirus-based RNAi system was developed to drive expression and delivery of HBV-specific short hairpin RNA (shRNA) in a mouse model for HBV replication. METHODS: Hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in the sera of the mice were analyzed by quantitative sandwich enzyme linked immunosorbent assay (ELISA) technique, hepatitis B core antigen (HBcAg) and HBsAg in the livers of the mice were detected by immunohistochemical assay, HBV DNA and HBV mRNA were measured by fluorogenic quantitative polymerase chain reaction (FQ PCR) and quantitative real-time PCR respectively. RESULTS: Co-injection of HBV plasmids together with the lentivirus targeting HBV shRNA induced an RNAi response. Secreted HBsAg was reduced by 89% in mouse serum, and HBeAg was also significantly inhibited, immunohistochemical detection of HBcAg or HBsAg in the liver tissues also revealed substantial reduction. Lentiviral mediated shRNA caused a significant suppression in the levels of viral mRNA and DNA synthesis compared to the control group. CONCLUSION: Lentivirus-based RNAi can be used to suppress HBV replication in vivo, it might become a potential therapeutic strategy for treating HBV and other viral infections. PMID- 19804650 TI - New journal selection for quantitative survey of infectious disease research: application for Asian trend analysis. AB - BACKGROUND: Quantitative survey of research articles, as an application of bibliometrics, is an effective tool for grasping overall trends in various medical research fields. This type of survey has been also applied to infectious disease research; however, previous studies were insufficient as they underestimated articles published in non-English or regional journals. METHODS: Using a combination of Scopus and PubMed, the databases of scientific literature, and English and non-English keywords directly linked to infectious disease control, we identified international and regional infectious disease journals. In order to ascertain whether the newly selected journals were appropriate to survey a wide range of research articles, we compared the number of original articles and reviews registered in the selected journals to those in the 'Infectious Disease Category' of the Science Citation Index Expanded (SCI Infectious Disease Category) during 1998-2006. Subsequently, we applied the newly selected journals to survey the number of original articles and reviews originating from 11 Asian countries during the same period. RESULTS: One hundred journals, written in English or 7 non-English languages, were newly selected as infectious disease journals. The journals published 14,156 original articles and reviews of Asian origin and 118,158 throughout the world, more than those registered in the SCI Infectious Disease Category (4,621 of Asian origin and 66,518 of the world in the category). In Asian trend analysis of the 100 journals, Japan had the highest percentage of original articles and reviews in the area, and no noticeable increase in articles was revealed during the study period. China, India and Taiwan had relatively large numbers and a high increase rate of original articles among Asian countries. When adjusting the publication of original articles according to the country population and the gross domestic product (GDP), Singapore and Taiwan were the most productive. CONCLUSION: A survey of 100 selected journals is more sensitive than the SCI Infectious Disease Category from the viewpoint of avoiding underestimating the number of infectious disease research articles of Asian origin. The survey method is applicable to grasp global trends in disease research, although the method may require further development. PMID- 19804651 TI - Sexual practices among unmarried adolescents in Tanzania. AB - BACKGROUND: Sexual activities are increasingly changing from the cultural point of view what they used to be. Knowledge of these practices among adolescents may be a basis to create awareness among adolescents on practices that involve risks. This study aims to assess sexual practices among unmarried adolescents in Tanzania. METHODS: A cross-sectional survey was conducted among in-school and out of-school but unmarried adolescents aged 10 to 19 in five locations in Tanzania. A questionnaire was used to collect information and to characterize sexual practices among these adolescents. RESULTS: About 32% of adolescents reported being sexually active; a higher proportion being males than females. The only inquired and reported sexual practices include vaginal sex, masturbation, oral and anal sex. About 15% of sexually active adolescents reported having multiple sexual partners. Significantly more males reported having multiple partners than females. Nearly 42% of sexually active adolescents reported having used a condom during most recent sexual act. Females reported older partners at first sexual act. CONCLUSION: Adolescents experience several sexual practices that include penetrative and non-penetrative. More males reported being sexually active than females. Despite adolescents reporting having multiple sexual partners, reported condom use during the most recent sexual act was low. We advocate for a more enhanced approach of reproductive health education that includes safer sex to adolescents without forgetting those in-schools. PMID- 19804652 TI - Demographic and circumstantial accounts of burn mortality in Cape Town, South Africa, 2001-2004: an observational register based study. AB - BACKGROUND: Burns are a persisting public health problem in low- and middle income countries; however, epidemiologic data for these settings is scarce. South Africa is no exception although there is an emerging knowledge base, especially for paediatric burns. The current study describes the epidemiology of burn mortality across the lifespan in Cape Town (2.9 million inhabitants in 2001), one of the six South African metropolitan centres. METHODS: The distribution of burn mortality across socio-demographic groups and also their circumstances of occurrence were investigated using four year (2001 to 2004) surveillance data from the National Injury Mortality Surveillance System (n = 1024 cases). RESULTS: Burn mortality occurred at a rate of 7.9 per 100,000 person-years (95% CI: 7.3 8.3). Males sustained fatal rates 2.2 times more than that for females (p < 0.001), with rates significantly higher in the 25 to 38 and 39 to 50 age groups than at other ages (p < 0.001). The greatest difference between male and female deaths was observed in the 25 to 38 year age group, when almost three male deaths occurred for every female one. The vast majority of fatal burns were registered as accidental and occurred in the home, either over the cold and wet months or during recreational periods over weekends and across the year. Alcohol intoxication was reported for the majority of those adults whose alcohol blood levels were tested (i.e. 52.6% of cases aged 16+ years). CONCLUSION: Besides paediatric burns, the high prevalence and circumstances of occurrence of burns among middle age men are a source of concern. There are reasons to believe that this over-representation is a reflection of detrimental living conditions, life style and poor socio-economic status. It is recommended that there be greater prioritisation of prevention activities that involve the control or management of kerosene heat sources, the provision of alternatives to flammable housing materials, and the implementation of strategies to reduce harmful drinking practices. PMID- 19804654 TI - The focused ion beam fold-out: sample preparation method for transmission electron microscopy. AB - We have developed the focused ion beam (FIB) fold-out technique, for transmission electron microscopy (TEM) sample preparation in which there is no fine polishing or dimpling, thus saving turnaround time. It does not require a nanomanipulator yet is still site specific. The sample wafer is cut to shape, polished down, and then placed in a FIB system. A tab containing the area of interest is created by ion milling and then "folded out" from the bulk sample. This method also allows a plan-view of the sample by removing material below the wafer's surface film or device near the polished edge. In the final step, the sample is thinned to electron transparency, ready to be analyzed in the TEM. With both a cross section and plan-view, our technique gives microscopists a powerful tool in analyzing multiple zone axes in one TEM session. The nature of the polished sample edge also includes the ability to sample many areas, allowing the user to examine a very large device or sample. More importantly, this technique could make multiple site-specific e-beam transparent specimens in one polished sample, which is difficult to do when prepared by other methods. PMID- 19804653 TI - Network of vascular diseases, death and biochemical characteristics in a set of 4,197 patients with type 1 diabetes (the FinnDiane Study). AB - BACKGROUND: Cardiovascular disease is the main cause of premature death in patients with type 1 diabetes. Patients with diabetic kidney disease have an increased risk of heart attack or stroke. Accurate knowledge of the complex inter dependencies between the risk factors is critical for pinpointing the best targets for research and treatment. Therefore, the aim of this study was to describe the association patterns between clinical and biochemical features of diabetic complications. METHODS: Medical records and serum and urine samples of 4,197 patients with type 1 diabetes were collected from health care centers in Finland. At baseline, the mean diabetes duration was 22 years, 52% were male, 23% had kidney disease (urine albumin excretion over 300 mg/24 h or end-stage renal disease) and 8% had a history of macrovascular events. All-cause mortality was evaluated after an average of 6.5 years of follow-up (25,714 patient years). The dataset comprised 28 clinical and 25 biochemical variables that were regarded as the nodes of a network to assess their mutual relationships. RESULTS: The networks contained cliques that were densely inter-connected (r > 0.6), including cliques for high-density lipoprotein (HDL) markers, for triglycerides and cholesterol, for urinary excretion and for indices of body mass. The links between the cliques showed biologically relevant interactions: an inverse relationship between HDL cholesterol and the triglyceride clique (r < -0.3, P < 10(-16)), a connection between triglycerides and body mass via C-reactive protein (r > 0.3, P < 10(-16)) and intermediate-density cholesterol as the connector between lipoprotein metabolism and albuminuria (r > 0.3, P < 10(-16)). Aging and macrovascular disease were linked to death via working ability and retinopathy. Diabetic kidney disease, serum creatinine and potassium, retinopathy and blood pressure were inter-connected. Blood pressure correlations indicated accelerated vascular aging in individuals with kidney disease (P < 0.001). CONCLUSION: The complex pattern of links between diverse characteristics and the lack of a single dominant factor suggests a need for multifactorial and multidisciplinary paradigms for the research, treatment and prevention of diabetic complications. PMID- 19804655 TI - Celebrating 40 years of energy dispersive X-ray spectrometry in electron probe microanalysis: a historic and nostalgic look back into the beginnings. AB - On February 2, 1968, R. Fitzgerald, K. Keil, and K.F.J. Heinrich published a seminal paper in Science (159, 528-530) in which they described a solid-state Si(Li) energy dispersive spectrometer (EDS) for electron probe microanalysis (EPMA) with, initially, a resolution of 600 eV. This resolution was much improved over previous attempts to use either gas-filled proportional counters or solid state devices for EDS to detect X-rays and was sufficient, for the first time, to make EDS a practically useful technique. It ushered in a new era not only in EPMA, but also in scanning electron microscopy, analytical transmission electron microscopy, X-ray fluorescence analysis, and X-ray diffraction. EDS offers many advantages over wavelength-dispersive crystal spectrometers, e.g., it has no moving parts, covers the entire X-ray energy range of interest to EPMA, there is no defocusing over relatively large distances across the sample, and, of particular interest to those who analyze complex minerals consisting of many elements, all X-ray lines are detected quickly and simultaneously. PMID- 19804656 TI - Impact of 40 years of technology advances on EDS system performance. AB - The rapid advance and falling costs of computing power and data storage during the last 40 years have greatly enhanced the data reduction speed and analytical capacity of energy dispersive spectroscopy (EDS) systems. At the same time, the solid state X-ray detector [until recently, the Si(Li) diode] has seen performance increases due to the use of advanced materials and processing techniques. In addition, the performance of the electronic components (field effect transistor, preamp, and pulse processor) of an EDS system has been constantly improved. These technology advances have resulted in improved spectral quality, excellent light element detection, and increased count rate performance. The results have been truly remarkable and driven by the needs of the analyst. This article will summarize the progress made in these areas in the last 40 years and make a brief reference to prospects for future development in EDS system performance. PMID- 19804657 TI - Spatial frequency components influence cell activity in the inferotemporal cortex. AB - We studied the correlation between the spatial frequency of complex stimuli and neuronal activity in the monkey inferotemporal (IT) cortex while performing a task that required visual recognition. Single-cell activity was recorded from the right IT cortex. The frequency components of the images used as stimuli were analyzed by using a fast Fourier transform, and a modulus was obtained for 40 spatial frequency ranges from 0.3 to 11.1 cycles/deg. We recorded 82 cells showing statistically significant responses (analysis of variance, P < 0.05) to at least one of the images used as a stimulus. Seventy-eight percent of these cells (n = 64) showed significant responses to at least three images, and in two thirds of them (n = 42), we found a statistically significant correlation (P < 0.05) between cell response and the modulus amplitude of at least one frequency range present in the images. Our results suggest that information about spatial frequency of the visual images is present in the IT cortex. PMID- 19804658 TI - Transmission routes and risk factors for autochthonous hepatitis E virus infection in Europe: a systematic review. AB - Increasing numbers of non-travel-associated hepatitis E virus (HEV) infections have been reported in Europe in recent years. Our objective was to review the evidence on risk factors and transmission routes of autochthonous HEV infection and hepatitis E in Europe in order to develop recommendations for future research, prevention and control. A systematic literature review was performed to identify all primary reports and studies published during 1998-2008 on hepatitis E in humans and animals in Europe by searching Pubmed, reference lists of major articles and international conference proceedings. Each of the 106 included studies was categorized into one of three evidence levels (EL) based on study design and diagnostic methodology. The evidence was generally weak (73 were assigned to EL1, two to both EL1 and EL2, and 30 to EL2), further compounded by the use of poorly validated serological assays in some studies. Only one case control study was assigned to EL3. Persons with autochthonous hepatitis E infection were on average older than the general population and predominantly male. There was no evidence for one main transmission route of HEV infection or risk factor for hepatitis E. However, zoonotic transmission seemed likely and person-to-person transmission too inefficient to cause clinical disease. Multiple routes of transmission probably exist and should be further investigated through analytical studies and reliable diagnostic kits. Based on current evidence that points to zoonotic transmission from pigs, thorough cooking of all porcine products, prevention of cross-contamination in the kitchen and improved education for occupationally exposed people (e.g. pig farmers, veterinarians and sewage workers) may help prevent HEV infection. Although evidence for parenteral transmission is limited, it is recommended that a risk assessment is undertaken. PMID- 19804678 TI - Explaining Alberta's rising mesothelioma rates. AB - Although mesothelioma rates have been rising worldwide, little is known about mesothelioma trends in Alberta. This population-based descriptive study used Alberta Cancer Board Registry data from 1980 to 2004 to develop an age-period cohort model of male pleural mesothelioma incidence rates over time. Both age and cohort effects are associated with incidence rates. The highest-risk cohort comprised men born between 1930 and 1939, reflecting widespread asbestos use and exposure beginning in the 1940s in Canada. We predict that 1393 Albertan men 40 years and older will die of pleural mesothelioma between 1980 and 2024; 783 (56.2%) of these deaths will occur between 2010 and 2024. The total number of mesothelioma deaths in Alberta will be higher when all age groups, both sexes, and all disease sites are included, with numbers likely peaking sometime between 2015 and 2019. In addition to the ongoing efforts that focus on eliminating asbestos-related disease in Alberta, the challenge is to implement surveillance systems to prevent future epidemics of preventable occupational cancers in Alberta. PMID- 19804679 TI - An intersectoral network for chronic disease prevention: the case of the Alberta healthy living network. AB - Chronic Diseases (CDs) are the leading causes of death and disability worldwide. CD experts have long promoted the use of integrated and intersectoral approaches to strengthen CD prevention efforts. This qualitative case study examined the perceived benefits and challenges associated with implementing an intersectoral network dedicated to CD prevention. Through interviewing key members of the Alberta Healthy Living Network (AHLN, or the Network), two overarching themes emerged from the data. The first relates to contrasting views on the role of the AHLN in relation to its actions and outcomes, especially concerning policy advocacy. The second focuses on the benefits and contributions of the AHLN and the challenge of demonstrating non-quantifiable outcomes. While the respondents agreed that the AHLN has contributed to intersectoral work in CD prevention in Alberta and to collaboration among Network members, several did not view this achievement as an end in itself and appealed to the Network to engage more in change-oriented activities. Managing contrasting expectations has had a significant impact on the functioning of the Network. PMID- 19804680 TI - Navigating the health care system: perceptions of patients with chronic pain. AB - A new framework is needed for patients with chronic pain and their primary care physicians that acknowledges the individual's experiences and provides evidence informed education and better linkages to community-based resources. This study describes the experience of 19 chronic-pain sufferers who seek relief via the health care system. Their experiences were recorded through in-depth semistructured interviews and analyzed through qualitative methods. The participants reported early optimism, then disillusionment, and finally acceptance of living with chronic pain. Both individuals with chronic pain and their health care professionals need evidence-informed resources and information on best practices to assist them to manage pain. Empathetic communication between health care professionals and individuals with chronic pain is crucial because insensitive communication negatively affects the individual, reduces treatment compliance and increases health care utilization. PMID- 19804681 TI - School-based tobacco-control programming and student smoking behaviour. AB - The study examined the association of a school-based tobacco-control program with students' smoking behaviour over time using three cross-sectional, provincial census datasets (grade 10 students in 1999, grade 11 students in 2000, grade 12 students in 2001). Data were collected from all secondary schools in Prince Edward Island (Canada) using the Tobacco module of the School Health Action, Planning and Evaluation System (SHAPES). The proportion of regular smokers increased from grade 10 (22.3%) to grade 12 (27.8%, chi(2) = 10.35, df = 1, p < 0.001). Being exposed to different school-based tobacco programs and policies in grades 10 and 11 was not associated with the smoking behaviour of grade 12 students. The strongest predictors of smoking behaviour were having friends or close family members who smoke. This preliminary evidence suggests that programs and policies associated with banning smoking and enforcing smoking restrictions at school may be insufficient unless they also address the influence of smoking peers and family members and link to comprehensive programming within the broader context of other community and policy level interventions. PMID- 19804682 TI - A deprivation index for health planning in Canada. AB - Administrative databases in the Canadian health sector do not contain socio economic information. To facilitate the monitoring of social inequalities for health planning, this study proposes a material and social deprivation index for Canada. After explaining the concept of deprivation, we describe the methodological aspects of the index and apply it to the example of premature mortality (i.e. death before the age of 75). We illustrate variations in deprivation and the links between deprivation and mortality nationwide and in different geographic areas including the census metropolitan areas (CMAs) of Toronto, Montreal and Vancouver; other CMAs; average-size cities, referred to as census agglomerations (CAs); small towns and rural communities; and five regions of Canada, namely Atlantic, Quebec, Ontario, the Prairies and British Columbia. Material and social deprivation and their links to mortality vary considerably by geographic area. We comment on the results as well as the limitations of the index and its advantages for health planning. PMID- 19804683 TI - Gemcitabine for the treatment of metastatic breast cancer. AB - This paper presents a summary of the evidence review group (ERG) report into the evidence for the clinical effectiveness and cost-effectiveness of gemcitabine with paclitaxel for the first-line treatment of metastatic breast cancer (MBC) in patients who have already received chemotherapy treatment with an anthracycline, compared with current standard of care, based upon the manufacturer's submission to the National Institute for Health and Clinical Excellence (NICE) as part of the single technology appraisal (STA) process. The clinical evidence for gemcitabine as a treatment for MBC comes from the unpublished JHQG trial (some data commercial-in-confidence): overall survival was 3 months longer for the gemcitabine/paclitaxel arm (18.5 months) than for the paclitaxel arm (15.8 months) (p = 0.0489); gemcitabine/paclitaxel also improved tumour response and time to documented progression of disease compared with paclitaxel monotherapy, but haematological serious adverse events were more common. In the absence of any formal methods of indirect comparison there is insufficient robust evidence to compare the relative effectiveness of gemcitabine/paclitaxel with docetaxel monotherapy or docetaxel/capecitabine combination therapy. The manufacturers used a Markov state transition model to estimate the effect of treatment with five different chemotherapy regimes, adopting a 3-year time horizon with docetaxel monotherapy as the comparator. Health state utilities for different stages of disease progression and for patients experiencing treatment-related toxicity are used to derive quality-adjusted life expectancy with each treatment. The base case cost-effectiveness estimate for gemcitabine/paclitaxel versus docetaxel is 17,168 pounds per quality-adjusted life-year (QALY). When longer survival with docetaxel is assumed in a sensitivity analysis, the incremental cost effectiveness ratio (ICER) is 30,000 pounds per QALY. Probabilistic sensitivity analysis estimates a 70% probability of gemcitabine/paclitaxel being cost effective relative to docetaxel at a willingness-to-pay threshold of 35,000 pounds. There is considerable uncertainty over the results because of the lack of formal quality assessment or assessment of the comparability of the 15 trials included in the input data, and the questionable validity of the indirect comparison method adopted. An illustrative analysis using a different method for indirect comparison carried out by the ERG produces an ICER of 45,811 pounds per QALY for gemcitabine/paclitaxel versus docetaxel. The guidance issued by NICE in November 2006 as a result of the STA states that gemcitabine in combination with paclitaxel, within its licensed indication, is recommended as an option for the treatment of MBC only when docetaxel monotherapy or docetaxel plus capecitabine is also considered appropriate. PMID- 19804684 TI - Varenicline in the management of smoking cessation: a single technology appraisal. AB - This paper presents a summary of the submission's evidence for the clinical effectiveness and cost-effectiveness of varenicline for smoking cessation included four studies of varenicline (one of which was commercial-in-confidence) and a meta-analysis of varenicline versus nicotine replacement therapy (NRT), bupropion and placebo. Two controlled trials of 12 weeks of varenicline versus sustained-release bupropion and placebo suggested that varenicline results in a statistically significant improvement in the odds of quitting at 12 weeks [odds ratio (OR) for quit rate during last 4 weeks of the study: 1.90-1.93 (p < 0.001) varenicline versus bupropion; 3.85 (p < 0.001) varenicline versus placebo). The ORs for sustained abstinence (weeks 9-52) for varenicline versus bupropion were 1.77 (p = 0.004) and 1.46 (p = 0.057), and for varenicline versus placebo were 2.66-3.09 (p < 0.01). A placebo-controlled maintenance trial examined whether a further 12 weeks of varenicline would maintain the rate of abstinence among those successfully treated on one 12-week course [OR = 2.48 at week 24 for varenicline versus placebo (p < 0.001)]. The meta-analysis suggested that varenicline was superior to placebo and bupropion at 1 year and 3 months. Based on indirect comparisons, varenicline was reported to be superior to NRT when compared with placebo or all controls at 1 year and 3 months. The submission presented a state transition model to estimate the incremental cost-effectiveness of varenicline compared with bupropion, NRT and placebo. The model suggests that varenicline dominates bupropion, NRT and placebo.Treatment efficacy was based on a pooled analysis of 1-year quit rates from the varenicline clinical trials. Assuming a willingness-to-pay threshold range of 20,000-30,000 pounds per quality-adjusted life-year gained, the probabilistic sensitivity analysis suggests that the probability that varenicline produces the greatest amount of net benefit is 0.70. Weaknesses of the manufacturer's submission include the assumption that only a single quit attempt using a single smoking cessation intervention is made, the presence of multiple computational errors and a limited sensitivity analysis. In conclusion, varenicline is likely to be clinically and cost-effective for smoking cessation assuming that each user makes a single quit attempt. The key area of uncertainty concerns the long-term experience of subjects who have remained abstinent from smoking beyond 12 months. The guidance issued by the National Institute for Health and Clinical Excellence in July 2007 states that varenicline is recommended within its licensed indications as an option for smokers who have expressed a desire to quit smoking and that varenicline should normally be prescribed only as part of a programme of behavioral support. PMID- 19804685 TI - Alteplase for the treatment of acute ischaemic stroke: a single technology appraisal. AB - This paper presents a summary of the evidence review group report into the clinical effectiveness and cost-effectiveness of alteplase for the treatment of acute ischaemic stroke, in accordance with the licensed indication, based upon the evidence submission from the manufacturer to the National Institute for Health and Clinical Excellence (NICE) as part of the single technology appraisal (STA) process. The submitted clinical evidence included several randomised controlled trials indicating that, in highly selected patients, alteplase administered at a licensed dose within 3 hours of the onset of acute ischaemic stroke is associated with a statistically significant reduction in the risk of death or dependency at 3 months compared with placebo, despite a significantly increased risk of symptomatic intracranial haemorrhage within the first 7-10 days. Data from the National Institute of Neurological Disorders and Stroke (NINDS) trial suggest that the benefit of treatment is sustained at 6 and 12 months. However, data from observational studies suggest that few patients with acute ischaemic stroke will be eligible for alteplase therapy under the terms of the current licensing agreement. In particular, many patients will be excluded by virtue of their age, and many more by the restriction of therapy to patients in whom treatment can be initiated within 3 hours of symptom onset. The manufacturer's submission included a state transition model evaluating the impact of treatment with alteplase within 3 hours of onset of stroke symptoms compared to standard treatment reporting that, in the base-case analysis, alteplase was both less costly and more effective than standard treatment. This increased to a maximum of approximately 4000 pounds upon one-way sensitivity analysis of the parameters. The probabilistic sensitivity analysis presented within the submission suggests that the probability that alteplase has a cost-effectiveness ratio greater than 20,000 pounds per quality-adjusted life-year (QALY) gained is close to 1 (0.99). The results of the short-term model demonstrate that alteplase is cost-effective over a 12-month period, with an incremental cost-effectiveness ratio of 14,026 pounds per QALY gained. This increased to a maximum of 50,000 pounds upon one-way sensitivity analysis of the parameters. At 12 months, the probabilistic sensitivity analysis presented within the submission suggests that the probability that alteplase has a cost-effectiveness ratio greater than 20,000 pounds per QALY gained is approximately 0.7. The guidance issued by NICE in April 2007 as a result of the STA states that alteplase is recommended for the treatment of acute ischaemic stroke only when used by physicians trained and experienced in the management of acute stroke and in centres with the required facilities. PMID- 19804686 TI - Rituximab for the treatment of rheumatoid arthritis. AB - This paper presents a summary of the evidence review group's critical review of the evidence for the clinical effectiveness and cost-effectiveness of rituximab for the treatment of severe rheumatoid arthritis (RA) following failure of previous therapy, including one or more tumour necrosis factor-alpha inhibitors (TNFi), compared with current standards of care, based upon the manufacturer's submission to the National Institute for Health and Clinical Excellence (NICE) as part of the single technology appraisal (STA) process. The submission's clinical evidence came from one randomised, placebo-controlled, double-blind trial (REFLEX -Random Evaluation of Long-term Efficacy of Rituximab in Rheumatoid Arthritis) comparing rituximab plus methotrexate (MTX) with placebo plus MTX in 517 patients with long-standing refractory RA. Rituximab plus MTX was more effective than placebo plus MTX across a range of primary and secondary outcome measures, e.g. American College of Rheumatology (ACR) responses, Health Assessment Questionnaire (HAQ). However, this evidence cannot be used directly to address the manufacturer's analysis of the decision problem because, in the REFLEX trial, rituximab was not compared with a relevant comparator (e.g. leflunomide or second or third TNFi). Long-term efficacy data for retreatment with rituximab are favourable, with an estimated mean time to retreatment of 307 days (n = 164). Evidence from a further five trials is presented as the basis for indirect comparisons with other disease-modifying antirheumatic drugs (DMARDs); however, it is not clear that all relevant clinical studies have been included in the indirect comparison exercise, the rationale for the choice of indirect comparison method adopted is unclear and the indirect comparison method used to adjust the ACR responses only uses a single value for the reference placebo. The submitted microsimulation Markov model was based upon the REFLEX trial. For the 'NICE recommended' scenario and the 'sequential TNFi' scenario, the original submission reports incremental cost-effectiveness ratios (ICERs) of 14,690 pounds and 11,601 pounds per quality-adjusted life-year (QALY) gained respectively. After model assumptions were adjusted to more realistic estimates by the ERG, the ICERs for the NICE-recommended scenario and the sequential use of TNFi range from 37,002 pounds to 80,198 pounds per QALY gained and from 28,553 pounds to 65,558 pounds per QALY gained respectively. The guidance issued by NICE in August 2007 states that rituximab in combination with methotrexate is recommended as an option for the treatment of adults with severe active rheumatoid arthritis who have had an inadequate response to or intolerance of other DMARDs including treatment with at least one TNFi therapy. PMID- 19804687 TI - Omalizumab for the treatment of severe persistent allergic asthma. AB - This paper presents a summary of the evidence review group (ERG) report into the clinical effectiveness and cost-effectiveness of omalizumab for the treatment of chronic severe persistent allergic asthma, in accordance with the licensed indication, based upon the evidence submission from Novartis to the National Institute for Health and Clinical Excellence (NICE) as part of the single technology appraisal (STA) process. The clinical evidence comes from a randomised controlled trial comparing omalizumab as an add-on to standard therapy with placebo and standard therapy over a 28-week treatment period. For the primary outcome of the rate of clinically significant asthma exacerbations, there was no statistically significant difference between treatment groups. However, after making a post hoc adjustment for a suggested 'clinically relevant' imbalance between trial arms in baseline exacerbation rate, the difference became marginally statistically significant. In terms of secondary outcomes, there were statistically significant differences favouring omalizumab over placebo in total emergency visits, Asthma Quality of Life Questionnaire scores, total symptom scores and lung function. Adverse events appeared to be similar between the trial arms. Results from three other publications are included in the manufacturer's submission as supporting evidence for the effectiveness of omalizumab, despite not meeting the inclusion criteria which adhere strictly to the licensed indication. The ERG checked and provided commentary on the manufacturer's model using standard checklists as well as undertook one-way sensitivity analysis, scenario analysis and a probabilistic sensitivity analysis. The cost effectiveness analysis estimates the incremental costs and consequences of omalizumab as an add-on to standard therapy. The base-case analysis of the trial's primary intention-to-treat population estimates a cost per quality adjusted life-year of 30,647 pounds. The ERG conducted one-way sensitivity analyses for parameters omitted from the manufacturer's submission sensitivity analysis. The results were most sensitive to variation in the utility values for omalizumab responders, and the unit cost of omalizumab. The guidance issued by NICE in November 2007 as a result of the STA states that omalizumab is recommended as a possible treatment for adults and young people over 12 years with severe persistent allergic asthma when their asthma meets certain conditions. Omalizumab treatment should be given along with the person's current asthma medicines. It should be prescribed by a doctor who is experienced in asthma and allergy medicine at a specialist centre. If omalizumab does not control the asthma after 16 weeks, treatment should be stopped. PMID- 19804688 TI - Rituximab for the treatment of relapsed or refractory stage III or IV follicular non-Hodgkin's lymphoma. AB - This paper presents a summary of the evidence review group report into the clinical effectiveness and cost-effectiveness of rituximab for the treatment of relapsed or refractory stage III or IV follicular non-Hodgkin's lymphoma (NHL), in accordance with the licensed indication, based upon the evidence submission from Roche Products Ltd to the National Institute for Health and Clinical Excellence (NICE) as part of the single technology appraisal (STA) process. The submitted clinical evidence included two randomised controlled trials [European Organisation for Research and Treatment of Cancer (EORTC) and German Low Grade Lymphoma Study Group - Fludarabine, Cyclophosphamide and Mitoxantrone and (GLSG FCM)] comparing the clinical effects of chemotherapy with or without rituximab in the induction of remission at first or second relapse and the clinical benefits of rituximab maintenance therapy versus the NHS's current clinical practice of observation for follicular lymphoma (FL) patients. Both trials showed that in patients with relapsed FL the addition of rituximab to chemotherapy induction treatment increased overall response rates. Furthermore, rituximab maintenance therapy increased the median length of remission when compared with observation only. Safety data from the two trials showed that while the majority of patients reported some adverse events, the number of patients withdrawing from treatment in the EORTC trial was low, with rates not being reported for the GLSG-FCM trial. The most commonly reported adverse events were blood/bone marrow toxicity, skin rashes and allergies. The ERG reran the manufacturer's economic model after altering several of the assumptions and parameter values in order to recalculate the cost-utility ratios, quality-adjusted life-years (QALYs) and estimates of benefits. The manufacturer reported that maintenance therapy with rituximab was cost-effective compared with observation against commonly applied thresholds, with an incremental cost-effectiveness ratio of 7721 pounds per QALY gained. The greatest clinical effectiveness is achieved by R-CHOP followed by rituximab maintenance (R-CHOP>R) and this treatment strategy had the greatest probability of being cost-effective for a QALY of approximately 18,000 pounds or greater. The guidance issued by NICE as a result of the STA states that in people with relapsed stage III or IV follicular NHL, rituximab is now an option in combination with chemotherapy to induce remission or alone as maintenance therapy during remission. Rituximab monotherapy is also an option for people with relapsed or refractory disease when all alternative treatment options have been exhausted. PMID- 19804689 TI - Adalimumab for the treatment of psoriasis. AB - This paper presents a summary of the evidence review group (ERG) report into the clinical and cost-effectiveness of adalimumab for the treatment of moderate to severe plaque psoriasis based upon a review of the manufacturer's submission to the National Institute for Health and Clinical Excellence (NICE) as part of the single technology appraisal (STA) process. The submission's clinical evidence came from three randomised controlled trials comparing adalimumab with placebo, two extension studies and one ongoing open-label extension study. The studies were of reasonable quality and measured a range of clinically relevant outcomes. A higher proportion of patients on 40 mg adalimumab every other week achieved an improvement on the Psoriasis Area and Severity Index (PASI) of at least 75% (PASI 75) compared with placebo groups after 12 or 16 weeks of treatment, and there was a statistically significant difference in favour of adalimumab for the proportion of patients achieving a PASI 50 and a PASI 90. In a mixed treatment comparison, for each PASI outcome the probability of a response was greater for infliximab than for adalimumab, but the probability of response with adalimumab was greater than that with etanercept, efalizumab and non-biological systemic therapies. Adverse event rates were similar in the treatment and placebo arms and discontinuations because of adverse events were low and comparable between groups. The submission's economic model presents treatment effectiveness for adalimumab versus other biological therapies based upon utility values obtained from two clinical trials. The model is generally internally consistent and appropriate to psoriasis in terms of structural assumptions and the methods used are appropriate. The base-case incremental cost-effectiveness ratio for adalimumab compared with supportive care for patients with severe psoriasis was 30,538 pounds per quality-adjusted life-year. Scenario analysis shows that the model was most sensitive to the utility values used. Weaknesses of the clinical evidence included not undertaking a systematic review of the comparator trials, providing very little in the way of a narrative synthesis of outcome data from the key trials and not performing a meta-analysis so that the overall treatment effect of adalimumab achieved across the trials is unknown. Weaknesses of the economic model included that the assumptions made to estimate the cost effectiveness of intermittent etanercept used inconsistent methodology for costs and benefits and there were no clear data on the amount of inpatient care required under supportive care. The NICE guidance issued as a result of the STA states that adalimumab is recommended as a treatment option for adults with plaque psoriasis in whom anti-tumour necrosis factor treatment is being considered and when the disease is severe and when the psoriasis has not responded to standard systemic therapies or the person is intolerant to or has a contraindication to these treatments. PMID- 19804690 TI - Dabigatran etexilate for the prevention of venous thromboembolism in patients undergoing elective hip and knee surgery: a single technology appraisal. AB - This paper presents a summary of the evidence review group (ERG) report into the clinical effectiveness and cost-effectiveness of dabigatran etexilate (DBG) for the prevention of venous thromboembolism (VTE) in patients undergoing elective hip and knee surgery based upon a review of the manufacturer's submission to the NICE as part of the single technology appraisal (STA) process. The submission's evidence came from three reasonable-quality trials comparing DBG with enoxaparin, and a comparison of DBG with fondaparinux based on the relative efficacy and safety as derived from a mixed treatment comparison (MTC) meta-analysis. DBG (220 mg and 150 mg once daily) is not inferior to enoxaparin (40 mg once daily and 30 mg twice daily) in terms of major VTE or VTE-related events (secondary outcome). Meta-analysis shows that 220 mg DBG is not inferior to enoxaparin (40 mg once daily or 30 mg twice daily) in reducing total VTE and all-cause mortality (primary outcome) in total hip or knee replacement, whereas there is uncertainty around the clinical effectiveness of 150 mg DBG for this outcome. In the MTC analysis DBG compared favourably with the other interventions, with the exception of extended enoxaparin and fondaparinux. The adverse event profile was not significantly different in those receiving DBG and those receiving enoxaparin. The submitted two-phase economic model compares DBG with enoxaparin and fondaparinux in total hip and knee replacement. The model structure is appropriate and the model assumptions are reasonable. The health states, costs, utilities and recurrence rates used are considered to be appropriate for the required analysis. The model estimated that at the licensed dose of 220 mg once daily DBG dominates enoxaparin in both total hip replacement and total knee replacement and that at the lower dose of 150 mg once daily DBG dominates enoxaparin in total hip replacement and enoxaparin dominates DBG in total knee replacement. DBG is less cost-effective than fondaparinux in total hip replacement at both doses; the cost per quality-adjusted life-year of fondaparinux versus DBG is 11,111 pounds and 6857 pounds for the higher and lower doses of DBG respectively. In total knee replacement, both DBG doses are dominated by fondaparinux. For DBG versus all comparators in all cases the cost effectiveness results are based on small incremental cost and health benefits. Weaknesses of the submitted evidence include that methods used for screening studies, data extraction and applying quality assessment criteria to included studies, as well as key details of trials included in the MTC, were not adequately described. In addition, some input parameters into the modelling process are incorrect. The ERG was unable to correct all of these mistakes and the impact on the model results is therefore unknown. The National Institute for Health and Clinical Excellence guidance issued as a result of the STA states that DBG is recommended as an option for the primary prevention of VTE events in adults who have undergone elective total hip or knee replacement surgery. PMID- 19804691 TI - Romiplostim for the treatment of chronic immune or idiopathic thrombocytopenic purpura: a single technology appraisal. AB - This paper presents a summary of the evidence review group (ERG) report into the clinical and cost-effectiveness of romiplostim for the treatment of adults with chronic immune or idiopathic thrombocytopenic purpura (ITP) based upon a review of the manufacturer's submission to the National Institute for Health and Clinical Excellence (NICE) as part of the single technology appraisal (STA) process. The submission's evidence came from two relatively high-quality randomised controlled trials (RCTs). The ERG found no evidence that any important data were missed or that data extraction was inaccurate. In both RCTs more patients in the romiplostim than in the placebo group achieved a durable platelet response [non-splenectomised patients: romiplostim 25/41 (61%), placebo 1/21 (5%), odds ratio (OR) 24.45, 95% confidence interval (CI) 3.34 to 179.18; splenectomised patients: romiplostim 16/42 (38%), placebo 0/21 (0%), OR 8.5 (95% CI 1.15 to 372)] and an overall platelet response [non-splenectomised patients: romiplostim 36/41 (88%), placebo 3/21 (14%), OR 34.74, 95% CI 7.77 to 155.38; splenectomised patients: romiplostim 33/42 (79%), placebo 0/21 (0%), OR 16.6 (95% CI 2.37 to 706]. The difference in mean period with a platelet response was 13.9 weeks (95% CI 10.5 to 17.4) in favour of romiplostim in the RCT of non splectomised patients and 12.1 weeks (95% CI 8.7 to 15.6) in favour of romiplostim in the RCT of splectomised patients. The manufacturer's economic model evaluated the cost-effectiveness of romiplostim compared with standard care. The ERG had concerns about the way the decision problem was addressed in the economic model and about the non-adjustment of findings for confounding factors. In non-splenectomised patients, using romiplostim as a first option treatment, the base-case incremental cost-effectiveness ratio (ICER) was 14,840 pounds per quality-adjusted life-year (QALY). In splenectomised patients the ICER was 14,655 pounds per QALY. Additional sensitivity analyses performed by the ERG identified two issues of importance: whether individuals entered the model on watch and rescue or on active therapy in the comparator arm (ICER 21,674 pounds per QALY for non-splenectomised patients, 29,771 pounds per QALY for splenectomised patients); whether it was assumed that any unused medicine would be wasted. Combining all of the separate sensitivity analyses, and assuming that watch and rescue was not the first-line treatment, increased the ICERs further (non-splenectomised 37,290 pounds per QALY; splenectomised 131,017 pounds per QALY). In conclusion, the manufacturer's submission and additional work conducted by the ERG suggest that romiplostim has short-term efficacy for the treatment of ITP, but there is no robust evidence on long-term effectiveness or cost effectiveness of romiplostim compared with relevant comparators. PMID- 19804692 TI - Sunitinib for the treatment of gastrointestinal stromal tumours: a critique of the submission from Pfizer. AB - The submission's evidence for the clinical effectiveness and cost-effectiveness of sunitinib for the treatment of gastrointestinal stromal tumours (GISTs) is based on a randomised controlled trial (RCT) comparing sunitinib with placebo for people with unresectable and/or metastatic GIST after failure of imatinib and with Eastern Cooperative Oncology Group (ECOG) progression status 0-1, and an ongoing, non-comparative cohort study of a similar population but with ECOG progression status 0-4. The searches are appropriate and include all relevant studies and the RCT is of high quality. In the RCT sunitinib arm overall survival was 73 median weeks [95% confidence interval (CI) 61 to 83] versus 75 median weeks (95% CI 68 to 84) for the cohort study. However, time to tumour progression in the cohort study was different from that in the RCT sunitinib arm [41 (95% CI 36 to 47) versus 29 (95% CI 22 to 41) median weeks respectively]. Median progression-free survival with sunitinib was 24.6 weeks (95% CI 12.1 to 28.4) versus 6.4 weeks (95% CI 4.4 to 10.0) on placebo (hazard ratio 0.333, 95% CI 0.238 to 0.467, p < 0.001). The manufacturer used a three-state Markov model to model the cost-effectiveness of sunitinib compared with best supportive care for GIST patients; the modelling approach and sources and justification of estimates are reasonable. The base-case incremental cost-effectiveness ratio (ICER) was 27,365 pounds per quality-adjusted life-year (QALY) with the first cycle of sunitinib treatment not costed; when we included the cost of the first treatment cycle we estimated a base-case ICER of 32,636 pounds per QALY. Pfizer's sensitivity analysis produced a range of ICERs from 15,536 pounds per QALY to 59,002 pounds per QALY. Weaknesses of the manufacturer's submission include that the evidence is based on only one published RCT; that 84% of the RCT control population crossed over to the intervention group, giving rise to the use of unusual rank preserved structural failure time (RPSFT) analysis to correct for possible bias; and that a number of errors and omissions were made in the probabilistic sensitivity analysis, meaning that it is not possible to come to firm conclusions about the cost-effectiveness of sunitinib for GIST in this patient population. In conclusion, during the blinded phase of the RCT, overall survival was significantly longer in the sunitinib arm than in the placebo arm (hazard ratio 0.491, 95% CI 0.290 to 0.831, p <0.007). However, intention-to treat analysis of the entire study showed no statistically significant difference in overall survival for those who received sunitinib (73 weeks) versus those who received placebo (65 weeks) (hazard ratio 0.876, 95% CI 0.679 to 1.129, p = 0.306). PMID- 19804693 TI - Contact urticaria due to phenoxyethanol in an aftershave. AB - The occurrence of pruritus immediately after application of an aftershave product is usually due to irritant contact dermatitis. We report a case of contact urticaria in a male patient, produced by an aftershave product containing phenoxyethanol. PMID- 19804694 TI - A mechanism-based classification of dermatologic reactions to biologic agents used in the treatment of cutaneous disease: Part 1. AB - Biologic therapies are an efficacious new method of controlling a number of chronic conditions. Data regarding these medications continues to emerge, giving clinicians a greater understanding of their side effects profiles. The biologic agents used in dermatology, particularly the tumor necrosis factor-alpha inhibitors, have a number of varied dermatologic side effects. In this two-part article, we perform a review of literature regarding the cutaneous side effects of infliximab, etanercept, adalimumab, rituximab, efalizumab, and alefacept. In Part 1, we will discuss cutaneous infections, malignancy, rebound phenomenon, eczema, atopic dermatitis, lichenoid reactions, granulomatous disease, pruritus, acne, and progressive multifocal leukoencephalopathy. PMID- 19804695 TI - Meta-analysis of genome-wide linkage studies of atopic dermatitis. AB - BACKGROUND: Atopic dermatitis (AD) is considered as a hereditary skin disease. Recently, a latest form of genetic research, genome-wide linkage study, has been carried out to detect its specific susceptibility genes. Since the results of these studies have been inconsistent, with linkage confirmed in different regions, we performed this meta-analysis. OBJECTIVE: To assess heritability and identify chromosomal regions showing evidence of AD. METHODS: A comprehensive and systematic search was conducted with PubMed databases. We combined results from three non-overlapping genome-wide linkage studies of AD, using the heterogeneity based genome scan meta-analysis (HEGESMA) method, a rank-based analysis that assesses the strongest evidence for linkage within bins of a 30-centimorgan width on autosomes and the X chromosome. RESULTS: The following ten bins had a p value of less than .05 in both weighted and unweighted analyses, suggesting that these bins contain AD-linked loci: 3.2, 3.4, 3.5, 4.5, 5.1, 5.2, 6.3, 9.2, 9.4, and 22.1. HEGESMA produced significant genome-wide evidence for linkage on 3p14.1 q12.3, with high average rank and low between-study heterogeneity; 5pter-p15.1 represents new significant loci not reported previously. CONCLUSION: HEGESMA confirms the region encoding CD80, CD86, and interleukins on chromosomes 3 and 5. The filaggrin gene (FLG) may be the susceptibility factor. PMID- 19804696 TI - Volatility of fragrance chemicals: patch testing implications. AB - BACKGROUND: Diagnostic and predictive patch testing to determine contact allergy due to fragrance materials requires applying a fixed dose of material to the skin. This dose can be affected by the volatile nature of fragrances; little data exist on how the loss of fragrance dose due to volatility affects patch testing. OBJECTIVE: (1) To evaluate pH dependence and evaporation rates of two fragrance chemicals, geraniol, citronellol, and a common fragrance solvent, diethyl phthalate (DEP) and (2) Assess implications for predictive patch-testing methods for fragrances. METHOD: pH analysis of each material at 1% for three values (4.0, 5.0, 7.0) was done over 40 hours. Volatility experiments for each material, nonradiolabeled and radiolabeled, were conducted over a 24-hour period, taking readings at six time points (5 minutes, 15 minutes, 40 minutes, 1 hour, 3 hours, and 24 hours). RESULTS: Evaporation rates were not sensitive to pH shifts from 4.0 to 7.0. Evaporation rates for nonradiolabeled materials were low: after 24 hours, geraniol lost 8.9%, citronellol 27.0% and DEP 14.5%. The volatility data for radiolabeled materials demonstrated that geraniol loses up to 39% of its dose, citronellol loses up to 26%, and DEP up to 14% within 40 minutes. CONCLUSION: The tendency of fragrance materials to evaporate can impact the dose being applied to the patch and therefore the result of the patch and ultimately the decision-making process regarding that fragrance material's safety. These data, developed with DEP, utilized in a predictive sensitization assay cannot be generalized. PMID- 19804697 TI - A 10-year retrospective study on palladium sensitivity. AB - BACKGROUND: Palladium has become an important contact allergen because of increased use in industry, jewelry, and dentistry. OBJECTIVE: To determine the frequency of palladium allergy in a US patch-test population tested to palladium. METHODS: A 10-year retrospective review (1997-2006) was performed on patients sensitive to palladium at the Department of Dermatology, Mayo Clinic, Rochester, MN. RESULTS: A total of 910 patients were tested to a series that included palladium chloride 2% in petrolatum. A palladium-positive patch-test result was noted in 110 patients (12.1%). Of the 106 patients sensitized to palladium who had records available for review, 15.1% had a diagnosis of lichen planus-like eruptions; 13.2%, burning mouth; 27.4%, stomatitis; and 29.2%, hand and body dermatitis. CONCLUSION: Palladium sensitivity is more common in oral diseases than in skin. We describe a previously unknown high degree of co-reactivity of gold with palladium to the same degree as the known co-reactivity with nickel. PMID- 19804698 TI - Rapid development of allergic contact dermatitis from dicyclohexylmethane-4,4' diisocyanate. AB - Isocyanates are widely used in the manufacturing of rigid and flexible foams, fibers, and coatings such as paints, varnishes, and elastomers but are rarely reported as contact sensitizers. The aliphatic diisocyanate dicyclohexylmethane 4,4'-diisocyanate (DMDI) is known to be a strong cutaneous sensitizer. We report a 27-year-old woman who developed a severe eczematous eruption on the arms, trunk, and abdomen within 2 weeks of employment at a company using the product Chem-Dec 808 isocyanate (containing 40-70% weight per weight DMDI). Patch testing was performed with the North American screening series, an isocyanate series, and 1% Chem-Dec 808 isocyanate. At day 7, a +++ reaction to Chem-Dec 808 isocyanate and a ++ reaction to the aromatic isocyanate diphenylmethane-4,4'-diisocyanate were observed. DMDI has strong sensitizing potential and requires rigorous safety monitoring in the workplace. PMID- 19804699 TI - Acrylates. PMID- 19804700 TI - Allergic contact dermatitis from retapamulin ointment. PMID- 19804701 TI - Contact dermatitis due to stockings. PMID- 19804703 TI - [Effect of apelin in cardiovascular system and water metabolism]. AB - Apelin is a recently identified peptide which plays an important role in cardiovascular function and in water metabolism. Apelin and its receptor APJ (apelin system) are involved in zebra fish and xenopus cardiovascular development. Apelin is a pro-angiogenic factor. Apelin is a potent cardiac inotropic agent and exerts a vasodilatory effect in arterial and venous territories. Apelin decreases vasopressin production and its plasma level is conversely related to plasma vasopressin. Several effects of the apelin system counteract those of the renin angiotensin system. It may represent a new and promising therapeutic opportunity for the treatment of cardiovascular diseases. PMID- 19804704 TI - [Angiotensin converting enzyme inhibitors and calcium antagonists: a synergistic action for a better prevention of cardiovascular events]. AB - Treatment of hypertension increasingly requires association of drugs. Among them, the association of calcium antagonists with angiotensin converting enzyme inhibitors is a new tempting approach since the two pharmaceutical classes have synergistic effects on blood pressure lowering and target organ protection whereas their effect is antagonistic for adverse events. Numerous large randomized clinical trials have validated this treatment strategy, and new fixed association are being developed. They apply to hypertensive patients, but also to coronary artery disease patients and more generally speaking to high cardiovascular risk subjects. PMID- 19804705 TI - [Budget impact analysis of Pradaxa thromboprophylaxis after total hip or total knee replacement]. AB - Venous thromboembolism is a serious disease that can be life-threatening in case of pulmonary embolism or induce major sequelae. Patients undergoing major orthopaedic surgery represent a population at high risk of venous thromboembolism. Recently Pradaxa has demonstrated its efficacy and safety versus enoxaparin in 2 pivotal trials. Beyond these clinical benefits on the treatment duration observed, the budget impact model's objective was to assess, from the French Sickness Funds perspective for retail market, the consequence of the introduction of this new drug on French setting for a theoretical, representative sample of 1,000 patients in each indication. The cost saving estimated by the model was in excess of 36,000 Euros per 1,000 procedures for total hip replacements. A univariate sensitivity analysis showed that the budget impact was robust and remained positive with the utilisation of Pradaxa in all cases tested. PMID- 19804706 TI - Collections of human biological samples for scientific purposes. Why do current regulation need to be clarified and how? AB - The collection of human biological samples is of major importance for future research in France and Europe. In recent years, new regulatory procedures have been designed to monitor these activities; but they are somewhat complex and some clarifications are needed. The law needs also to be amended. The definition of biobanking activities should be clarified, and regulatory procedures, including consultation of the Ethics Committee, declarations to the Ministry of Research and the protection of personal data, should be simplified. It is also of great importance to correctly define the modalities in which Biobanks are granted their authorisations. The role of Ethics Committees regarding the evaluation of information and the consent procedures should also be clarified, particularly when samples from children are used, or when the samples are used for genetic analyses. As well as scientific and public health aspects, the storage of human biological samples may also have important economic consequences. It is hence crucial to adapt the procedure for submitting patents, particularly when several public or private partners are working together. The possible changes to both French and European laws planned in the next months would be an ideal time to introduce these changes. PMID- 19804707 TI - [Comparison between analytic and anamnestic data about drug consumption among opiate addicts with substitutes therapy. A feasibility study]. AB - In this feasibility study of an observational nature, we used the protocol 'AnalyTox-Op' to compare analytical data with anamnestic reports gathered from specialized drug addiction treatment centers. These data were collected from 32 drug addicts who were patients undergoing treatment with addictive drug substitutes or prescribed psychotropic drugs. Urine toxicology screens were performed using immunological methods followed by confirmation with more specific techniques, i.e. gas chromatography coupled with mass spectrometry (GC-MS) and liquid chromatography with diode array detection (HPLC-DB). While complete agreement between patient reports of drug consumption (obtained from a questionnaire) and analytical data was only observed in 13 out of the 32 cases, a very good concordance was seen with opiate substitutes (88% with methadone and high-dosage buprenorphine, combined) and legally prescribed psychotropic drugs. Of note, however, was the omission of illicit drug use in patient questionnaires, especially with cocaine (22%) and recreational opiates (22%), causing discordance in the comparison. This study is currently being expanded to include a large number of participants across the country with the aim of obtaining data under homogeneous conditions, verifying the discordance we found and determining its significance. PMID- 19804708 TI - [Recourse to psychotropic medication in the Nord - Pas-de-Calais Region (France)]. AB - The prescriptions for psychoactive drugs presented to the Health Insurance system for reimbursement of charges are a source of information about some psychological illnesses and disorders in the context of the overuse of these drugs in France. In Nord - Pas-de-Calais Region, out of a total of 5 070 160 prescriptions in 2007, 576 493 individuals benefited from at least one such drug. Over the period of the study, 15.6% of the population of the region covered took at least one psychotropic medication. The rate of use was 11.7% for benzodiazepines, 7.6% for anti-depressants, 1.8% for anti-psychotics, 0.5% for treatment of alcohol dependence and 0.3% for opiate substitution therapy (OST). For the first three of these classes rate of use increased steadily with age. It was invariably much higher in women than in men. Men were treated more frequently for alcoholism and heroin dependency; the proportion taking medication initially increased with age (up to 40-49 years for alcohol dependency and 30-39 years for OST), before falling thereafter. PMID- 19804709 TI - Guidelines for submitting adverse event reports for publication. AB - Publication of case reports describing suspected adverse effects of drugs and medical products that include herbal and complementary medicines, vaccines and other biologicals and devices is important for postmarketing surveillance. Publication lends credence to important signals raised in these adverse event reports. Unfortunately, deficiencies in vital information in published cases can often limit the value of such reports by failing to provide enough details for either (i) a differential diagnosis or provisional assessment of cause-effect association, or (ii) a reasonable pharmacological or biological explanation. Properly described, a published report of one or more adverse events can provide a useful signal of possible risks associated with the use of a drug or medical product which might warrant further exploration. A review conducted by the Task Force authors found that many major journals have minimal requirements for publishing adverse event reports and some have none at all. Based on a literature review and our collective experience in reviewing adverse event case reports in regulatory, academic and industry settings, we have identified information that we propose should always be considered for inclusion in a report submitted for publication. These guidelines have been endorsed by the International Society for Pharmacoepidemiology (ISPE) and the International Society of Pharmacovigilance (ISoP) and are freely available on the societies' web sites. Their widespread distribution is encouraged. ISPE and ISoP urge biomedical journals to adopt these guidelines and apply them to case reports submitted for publication. They also encourage schools of medicine, pharmacy, and nursing to incorporate them into the relevant curricula that address the detection, evaluation and reporting of suspected drug or other medical product adverse events. PMID- 19804710 TI - Regaining eukaryotic identity after cell division: relax, young cell. PMID- 19804711 TI - Stochastic collective movement of cells and fingering morphology: no maverick cells. AB - The classical approach to model collective biological cell movement is through coupled nonlinear reaction-diffusion equations for biological cells and diffusive chemicals that interact with the biological cells. This approach takes into account the diffusion of cells, proliferation, death of cells, and chemotaxis. Whereas the classical approach has many advantages, it fails to consider many factors that affect multicell movement. In this work, a multiscale approach, the Glazier-Graner-Hogeweg model, is used. This model is implemented for biological cells coupled with the finite element method for a diffusive chemical. The Glazier-Graner-Hogeweg model takes the biological cell state as discrete and allows it to include cohesive forces between biological cells, deformation of cells, following the path of a single cell, and stochastic behavior of the cells. Where the continuity of the tissue at the epidermis is violated, biological cells regenerate skin to heal the wound. We assume that the cells secrete a diffusive chemical when they feel a wounded region and that the cells are attracted by the chemical they release (chemotaxis). Under certain parameters, the front encounters a fingering morphology, and two fronts progressing against each other are attracted and correlated. Cell flow exhibits interesting patterns, and a drift effect on the chemical may influence the cells' motion. The effects of a polarized substrate are also discussed. PMID- 19804712 TI - Microfilament orientation constrains vesicle flow and spatial distribution in growing pollen tubes. AB - The dynamics of cellular organelles reveals important information about their functioning. The spatio-temporal movement patterns of vesicles in growing pollen tubes are controlled by the actin cytoskeleton. Vesicle flow is crucial for morphogenesis in these cells as it ensures targeted delivery of cell wall polysaccharides. Remarkably, the target region does not contain much filamentous actin. We model the vesicular trafficking in this area using as boundary conditions the expanding cell wall and the actin array forming the apical actin fringe. The shape of the fringe was obtained by imposing a steady state and constant polymerization rate of the actin filaments. Letting vesicle flux into and out of the apical region be determined by the orientation of the actin microfilaments and by exocytosis was sufficient to generate a flux that corresponds in magnitude and orientation to that observed experimentally. This model explains how the cytoplasmic streaming pattern in the apical region of the pollen tube can be generated without the presence of actin microfilaments. PMID- 19804713 TI - Differential regulation of action potentials by inactivating and noninactivating BK channels in rat adrenal chromaffin cells. AB - Large-conductance Ca(2+)-activated K(+) (BK) channels can regulate cellular excitability in complex ways because they are able to respond independently to two distinct cellular signals, cytosolic Ca(2+) and membrane potential. In rat chromaffin cells (RCC), inactivating BK(i) and noninactivating (BK(s)) channels differentially contribute to RCC action potential (AP) firing behavior. However, the basis for these differential effects has not been fully established. Here, we have simulated RCC action potential behavior, using Markovian models of BK(i) and BK(s) current and other RCC currents. The analysis shows that BK current influences both fast hyperpolarization and afterhyperpolarization of single APs and that, consistent with experimental observations, BK(i) current facilitates repetitive firing of APs, whereas BK(s) current does not. However, the key functional difference between BK(i) and BK(s) current that accounts for the differential firing is not inactivation but the more negatively shifted activation range for BK(i) current at a given [Ca(2+)]. PMID- 19804714 TI - Modeling cardiac action potential shortening driven by oxidative stress-induced mitochondrial oscillations in guinea pig cardiomyocytes. AB - Ischemia-induced shortening of the cardiac action potential and its heterogeneous recovery upon reperfusion are thought to set the stage for reentrant arrhythmias and sudden cardiac death. We have recently reported that the collapse of mitochondrial membrane potential (DeltaPsi(m)) through a mechanism triggered by reactive oxygen species (ROS), coupled to the opening of sarcolemmal ATP sensitive potassium (K(ATP)) channels, contributes to electrical dysfunction during ischemia-reperfusion. Here we present a computational model of excitation contraction coupling linked to mitochondrial bioenergetics that incorporates mitochondrial ROS-induced ROS release with coupling between the mitochondrial energy state and electrical excitability mediated by the sarcolemmal K(ATP) current (I(K,ATP)). Whole-cell model simulations demonstrate that increasing the fraction of oxygen diverted from the respiratory chain to ROS production triggers limit-cycle oscillations of DeltaPsi(m), redox potential, and mitochondrial respiration through the activation of a ROS-sensitive inner membrane anion channel. The periods of transient mitochondrial uncoupling decrease the cytosolic ATP/ADP ratio and activate I(K,ATP), consequently shortening the cellular action potential duration and ultimately suppressing electrical excitability. The model simulates emergent behavior observed in cardiomyocytes subjected to metabolic stress and provides a new tool for examining how alterations in mitochondrial oxidative phosphorylation will impact the electrophysiological, contractile, and Ca(2+) handling properties of the cardiac cell. Moreover, the model is an important step toward building multiscale models that will permit investigation of the role of spatiotemporal heterogeneity of mitochondrial metabolism in the mechanisms of arrhythmogenesis and contractile dysfunction in cardiac muscle. PMID- 19804715 TI - Actin-myosin viscoelastic flow in the keratocyte lamellipod. AB - The lamellipod, the locomotory region of migratory cells, is shaped by the balance of protrusion and contraction. The latter is the result of myosin generated centripetal flow of the viscoelastic actin network. Recently, quantitative flow data was obtained, yet there is no detailed theory explaining the flow in a realistic geometry. We introduce models of viscoelastic actin mechanics and myosin transport and solve the model equations numerically for the flat, fan-shaped lamellipodial domain of keratocytes. The solutions demonstrate that in the rapidly crawling cell, myosin concentrates at the rear boundary and pulls the actin network inward, so the centripetal actin flow is very slow at the front, and faster at the rear and at the sides. The computed flow and respective traction forces compare well with the experimental data. We also calculate the graded protrusion at the cell boundary necessary to maintain the cell shape and make a number of other testable predictions. We discuss model implications for the cell shape, speed, and bi-stability. PMID- 19804716 TI - Low-affinity Ca2+ indicators compared in measurements of skeletal muscle Ca2+ transients. AB - The low-affinity fluorescent Ca(2+) indicators OGB-5N, Fluo-5N, fura-5N, Rhod-5N, and Mag-fluo-4 were evaluated for their ability to accurately track the kinetics of the spatially averaged free Ca(2+) transient (Delta[Ca(2+)]) in skeletal muscle. Frog single fibers were injected with one of the above indicators and, usually, furaptra (previously shown to rapidly track Delta[Ca(2+)]). In response to an action potential, the full duration at half-maximum of the indicator's fluorescence change (DeltaF) was found to be larger with OGB-5N, Fluo-5N, fura 5N, and Rhod-5N than with furaptra; thus, these indicators do not track Delta[Ca(2+)] with kinetic fidelity. In contrast, the DeltaF time course of Mag fluo-4 was identical to furaptra's; thus, Mag-fluo-4 also yields reliable kinetic information about Delta[Ca(2+)]. Mag-fluo-4's DeltaF has a larger signal/noise ratio than furaptra's (for similar indicator concentrations), and should thus be more useful for tracking Delta[Ca(2+)] in small cell volumes. However, because the resting fluorescence of Mag-fluo-4 probably arises largely from indicator that is bound with Mg(2+), the amplitude of the Mag-fluo-4 signal, and its calibration in Delta[Ca(2+)] units, is likely to be more sensitive to variations in [Mg(2+)] than furaptra's. PMID- 19804717 TI - Contribution of the cytoskeleton to the compressive properties and recovery behavior of single cells. AB - The cytoskeleton is known to play an important role in the biomechanical nature and structure of cells, but its particular function in compressive characteristics has not yet been fully examined. This study focused on the contribution of the main three cytoskeletal elements to the bulk compressive stiffness (as measured by the compressive modulus), volumetric or apparent compressibility changes (as further indicated by apparent Poisson's ratio), and recovery behavior of individual chondrocytes. Before mechanical testing, cytochalasin D, acrylamide, or colchicine was used to disrupt actin microfilaments, intermediate filaments, or microtubules, respectively. Cells were subjected to a range of compressive strains and allowed to recover to equilibrium. Analysis of the video recording for each mechanical event yielded relevant compressive properties and recovery characteristics related to the specific cytoskeletal disrupting agent and as a function of applied axial strain. Inhibition of actin microfilaments had the greatest effect on bulk compressive stiffness ( approximately 50% decrease compared to control). Meanwhile, intermediate filaments and microtubules were each found to play an integral role in either the diminution (compressibility) or retention (incompressibility) of original cell volume during compression. In addition, microtubule disruption had the largest effect on the "critical strain threshold" in cellular mechanical behavior (33% decrease compared to control), as well as the characteristic time for recovery ( approximately 100% increase compared to control). Elucidating the role of the cytoskeleton in the compressive biomechanical behavior of single cells is an important step toward understanding the basis of mechanotransduction and the etiology of cellular disease processes. PMID- 19804718 TI - Neurite branch retraction is caused by a threshold-dependent mechanical impact. AB - Recent results indicate that, in addition to chemical cues, mechanical stimuli may also impact neuronal growth. For instance, unlike most other cell types, neurons prefer soft substrates. However, the mechanisms responsible for the neuronal affinity for soft substrates have not yet been identified. In this study, we show that, in vitro, neurons continuously probe their mechanical environment. Growth cones visibly deform substrates with a compliance commensurate with their own. To understand the sensing of stiff substrates by growth cones, we investigated their precise temporal response to well-defined mechanical stress. When the applied stress exceeded a threshold of 274 +/- 41 pN/microm(2), neurons retracted and re-extended their processes, thereby enabling exploration of alternative directions. A calcium influx through stretch-activated ion channels and the detachment of adhesion sites were prerequisites for this retraction. Our data illustrate how growing neurons may detect and avoid stiff substrates--as a mechanism involved in axonal branch pruning--and provide what we believe is novel support of the idea that mechanics may act as guidance cue for neuronal growth. PMID- 19804719 TI - Formation of the nuclear envelope permeability barrier studied by sequential photoswitching and flux analysis. AB - In higher eukaryotes, the nuclear envelope breaks down during mitosis. It reforms during telophase, and nuclear import is reestablished within <10 min after anaphase onset. It is widely assumed that import functionality simultaneously leads to the exclusion of bulk cytoplasmic proteins. However, nuclear pore complex assembly is not fully completed when import capacity is regained, which raises the question of whether the transport and permeability barrier functions of the nuclear envelope are indeed coupled. In this study, we therefore analyzed the reestablishment of the permeability barrier of the nuclear envelope after mitosis in living cells by monitoring the flux of the reversibly photoswitchable fluorescent protein Dronpa from the cytoplasm into the nucleus after photoactivation. We performed many consecutive flux measurements in the same cell to directly monitor changes in nuclear envelope permeability. Our measurements at different time points after mitosis in individual cells show that contrary to the general view and despite the rapid reestablishment of facilitated nuclear import, the nuclear envelope remains relatively permeable for passive diffusion for the first 2 h after mitosis. Our data demonstrate that reformation of the permeability barrier of nuclear pore complexes occurs only gradually and is uncoupled from regaining active import functionality. PMID- 19804720 TI - Understanding ion conductance on a molecular level: an all-atom modeling of the bacterial porin OmpF. AB - All-atom molecular dynamics simulations of the ion current through OmpF, the major porin in the outer membrane of Escherichia coli, were performed. Starting from the crystal structure, the all-atom modeling allows us to calculate a parameter-free ion conductance in semiquantitative agreement with experiment. Discrepancies between modeling and experiment occur, e.g., at salt concentrations above 1 M KCl or at high temperatures. At lower salt concentrations, the ions have separate pathways along the channel surface. The constriction zone in the channel contains, on one side, a series of positively charges (R42, R82, R132), and on the opposite side, two negatively charged residues (D113, E117). Mutations generated in the constriction zone by removing cationic residues enhance the otherwise small cation selectivity, whereas removing the anionic residues reverses the selectivity. Reduction of the negatively charged residues decreases the conductance by half, whereas cationic residues enhance the conductance. Experiments on mutants confirm the results of the molecular-level simulations. PMID- 19804721 TI - Ca2+-calmodulin increases RyR2 open probability yet reduces ryanoid association with RyR2. AB - We have shown that physiological levels of Ca(2+)-calmodulin (Ca(2+)CaM; 50-100 nM) activate cardiac ryanodine receptors (RyR2) incorporated into bilayers and increase the frequency of Ca(2+) sparks and waves in cardiac cells. In contrast, it is well known that Ca(2+)CaM inhibits [(3)H]ryanodine binding to cardiac sarcoplasmic reticulum. Since the [(3)H]ryanodine binding technique does not reflect the effects of Ca(2+)CaM on RyR2 open probability (Po), we have investigated, using the reversible ryanoid, ryanodol, whether Ca(2+)CaM can directly influence the binding of ryanoids to single RyR2 channels independently of Po. We demonstrate that Ca(2+)CaM reduces the rate of ryanodol association to RyR2 without affecting the rate of dissociation. We also find that ryanodol-bound channels fluctuate between at least two distinct subconductance states, M(1) and M(2), in a voltage-dependent manner. Ca(2+)CaM significantly alters the equilibrium between these two states. The results suggest that Ca(2+)CaM binding to RyR2 causes a conformation change to regions of the channel that include the ryanoid binding site, thereby leading to a decrease in ryanoid association rate and modulation of gating within the ryanoid/RyR2 bound state. Our data provide a possible explanation for why the effects of Ca(2+)CaM at the single-channel level are not mirrored by [(3)H]ryanodine binding studies. PMID- 19804722 TI - Arginine-rich peptides destabilize the plasma membrane, consistent with a pore formation translocation mechanism of cell-penetrating peptides. AB - Recent molecular-dynamics simulations have suggested that the arginine-rich HIV Tat peptides translocate by destabilizing and inducing transient pores in phospholipid bilayers. In this pathway for peptide translocation, Arg residues play a fundamental role not only in the binding of the peptide to the surface of the membrane, but also in the destabilization and nucleation of transient pores across the bilayer. Here we present a molecular-dynamics simulation of a peptide composed of nine Args (Arg-9) that shows that this peptide follows the same translocation pathway previously found for the Tat peptide. We test experimentally the hypothesis that transient pores open by measuring ionic currents across phospholipid bilayers and cell membranes through the pores induced by Arg-9 peptides. We find that Arg-9 peptides, in the presence of an electrostatic potential gradient, induce ionic currents across planar phospholipid bilayers, as well as in cultured osteosarcoma cells and human smooth muscle cells. Our results suggest that the mechanism of action of Arg-9 peptides involves the creation of transient pores in lipid bilayers and cell membranes. PMID- 19804723 TI - Areas of monounsaturated diacylphosphatidylcholines. AB - We have studied the structural properties of monounsaturated diacylphosphatidylcholine lipid bilayers (i.e., diCn:1PC, where n = 14, 16, 18, 20, 22, and 24 is the number of acyl chain carbons). High-resolution x-ray scattering data were analyzed in conjunction with contrast-varied neutron scattering data using a technique we recently developed. Analyses of the data show that the manner by which bilayer thickness increases with increasing n in monounsaturated diacylphosphatidylcholines is dependent on the double bond's position. For commonly available monounsaturated diacylphosphatidylcholines, this results in the nonlinear behavior of both bilayer thickness and lipid area, whereas for diC18:1PC bilayers, lipid area assumes a maximum value. It is worthwhile to note that compared to previous data, our results indicate that lipid areas are smaller by approximately 10%. This observation highlights the need to revisit lipid areas, as they are often used in comparisons with molecular dynamics simulations. Moreover, simulators are encouraged to compare their results not only to x-ray scattering data, but to neutron data as well. PMID- 19804724 TI - Detergent-like activity and alpha-helical structure of warnericin RK, an anti Legionella peptide. AB - Warnericin RK is the first antimicrobial peptide known to be active against Legionella pneumophila, a pathogen bacterium that is responsible for severe pneumonia. Strikingly, this peptide displays a very narrow range of antimicrobial activity, almost limited to the Legionella genus, and a hemolytic activity. A similar activity has been described for delta-lysin, a well-known hemolytic peptide of Staphylococci that has not been described as antimicrobial. In this study we aimed to understand the mode of action of warnericin RK and to explain its particular target specificity. We found that warnericin RK permeabilizes artificial membranes in a voltage-independent manner. Osmotic protection experiments on erythrocytes showed that warnericin RK does not form well-defined pores, suggesting a detergent-like mode of action, as previously described for delta-lysin at high concentrations. Warnericin RK also permeabilized Legionella cells, and these cells displayed a high sensitivity to detergents. Depending on the detergent used, Legionella was from 10- to 1000-fold more sensitive than the other bacteria tested. Finally, the structure of warnericin RK was investigated by means of circular dichroism and NMR spectroscopy. The peptide adopted an amphiphilic alpha-helical structure, consistent with the proposed mode of action. We conclude that the specificity of warnericin RK toward Legionella results from both the detergent-like mode of action of the peptide and the high sensitivity of these bacteria to detergents. PMID- 19804725 TI - Simulation studies of stratum corneum lipid mixtures. AB - We present atomistic molecular dynamics results for fully hydrated bilayers composed of ceramide NS-24:0, free fatty acid 24:0 and cholesterol, to address the effect of the different components in the stratum corneum (the outermost layer of skin) lipid matrix on its structural properties. Bilayers containing ceramide molecules show higher in-plane density and hence lower rate of passive transport compared to phospholipid bilayers. At physiological temperatures, for all composition ratios explored, the lipids are in a gel phase with ordered lipid tails. However, the large asymmetry in the lengths of the two tails of the ceramide molecule leads to a fluidlike environment at the bilayer midplane. The lateral pressure profiles show large local variations across the bilayer for pure ceramide or any of the two-component mixtures. Close to the skin composition ratio, the lateral pressure fluctuations are greatly suppressed, the ceramide tails from the two leaflets interdigitate significantly, the depression in local density at the interleaflet region is lowered, and the bilayers have lowered elastic moduli. This indicates that the observed composition ratio in the stratum corneum lipid layer is responsible for both the good barrier properties and the stability of the lipid structure against mechanical stresses. PMID- 19804726 TI - Analysis of the interaction of the nucleotide base with myosin and the effect on substrate efficacy. AB - A wide variety of purine- and pyrimidine-based nucleotides can serve as a substrate for actomyosin mechanics, but with varying effectiveness. To understand the myosin-ATP interaction and in particular, the interactions with the base, we have used molecular dynamics simulations to model the interactions of myosin with ATP, CTP, UTP, aza-ATP, ITP, and GTP (in decreasing order of effectiveness as a substrate for the generation of motility) docked at the active site. The simulations with ATP, and x-ray structures, show a triad of conserved amino acids lining the nucleotide site that form a cyclical chain of nucleotide-protein hydrogen bonding interactions: ATP --> Y135 --> Y116 --> N188 --> ATP. Mechanical efficacy of a substrate correlates with its ability to maintain this coordination. Simulations modeling the active site of other myosin isoforms with different amino acids in the triad likewise imply that the amino acid composition at the nucleotide site could modulate function. The modeling has predictive power. In silico mutation experiments suggest mutations that would enhance GTP as a substrate for myosin while simultaneously making ATP a less effective substrate. PMID- 19804727 TI - Ryanodine receptor luminal Ca2+ regulation: swapping calsequestrin and channel isoforms. AB - Sarcoplasmic reticulum (SR) Ca(2+) release in striated muscle is mediated by a multiprotein complex that includes the ryanodine receptor (RyR) Ca(2+) channel and the intra-SR Ca(2+) buffering protein calsequestrin (CSQ). Besides its buffering role, CSQ is thought to regulate RyR channel function. Here, CSQ dependent luminal Ca(2+) regulation of skeletal (RyR1) and cardiac (RyR2) channels is explored. Skeletal (CSQ1) or cardiac (CSQ2) calsequestrin were systematically added to the luminal side of single RyR1 or RyR2 channels. The luminal Ca(2+) dependence of open probability (Po) over the physiologically relevant range (0.05-1 mM Ca(2+)) was defined for each of the four RyR/CSQ isoform pairings. We found that the luminal Ca(2+) sensitivity of single RyR2 channels was substantial when either CSQ isoform was present. In contrast, no significant luminal Ca(2+) sensitivity of single RyR1 channels was detected in the presence of either CSQ isoform. We conclude that CSQ-dependent luminal Ca(2+) regulation of single RyR2 channels lacks CSQ isoform specificity, and that CSQ dependent luminal Ca(2+) regulation in skeletal muscle likely plays a relatively minor (if any) role in regulating the RyR1 channel activity, indicating that the chief role of CSQ1 in this tissue is as an intra-SR Ca(2+) buffer. PMID- 19804728 TI - Molecular dynamics simulations of DNA-polycation complex formation. AB - Complexes formed from DNA and polycations are of interest because of their potential use in gene therapy; however, there remains a lack of understanding of the structure and formation of DNA-polycation complexes at atomic scale. In this work, molecular dynamics simulations of the DNA duplex d(CGCGAATTCGCG) in the presence of polycation chains are carried out to shed light on the specific atomic interaction that result in complex formation. The structures of complexes formed from DNA with polyethylenimine, which is considered one of the most promising DNA vector candidates, and a second polycation, poly-L-lysine, are compared. After an initial separation of approximately 50 A, the DNA and polycation come together and form a stable complex within 10 ns. The DNA does not undergo any major structural changes on complexation and remains in the B-form. In the formed complex, the charged amine groups of the polycation mainly interact with DNA phosphate groups, with polycation intrusion into the major and minor grooves dependent on the identity and charge state of the polycation. The ability of the polycation to effectively neutralize the charge of the DNA phosphate groups and the resulting influence on the DNA helix interaction are discussed. PMID- 19804729 TI - Power-laws in interferon-B mRNA distribution in virus-infected dendritic cells. AB - Interferon-beta (IFNB1) mRNA shows very large cell-to-cell variability in primary human dendritic cells infected by Newcastle disease virus, with copy numbers varying from a few to several thousands. Analysis of data from the direct measurement of the expression of this gene in its natural chromatin environment in primary human cells shows that the distribution of mRNA across cells follows a power law with an exponent close to -1, and thus encompasses a range of variation much more extensive than a Gaussian. We also investigate the single cell levels of IFNB1 mRNA induced by infection with Texas influenza A mutant viruses, which vary in their capacity to inhibit the signaling pathways responsible for activation of this gene. Here as well we observe power-law behavior for the distribution of IFNB1 mRNA, albeit over a truncated range of values, with exponents similar to the one for cells infected by Newcastle disease virus. We propose a model of stochastic enhanceosome and preinitiation complex formation that incorporates transcriptional pulsing. Analytical and numerical results show good agreement with the observed power laws, and thus support the existence of transcriptional pulsing of an unmodified, intact gene regulated by a natural stimulus. PMID- 19804730 TI - Effect of noise on DNA sequencing via transverse electronic transport. AB - Previous theoretical studies have shown that measuring the transverse current across DNA strands while they translocate through a nanopore or channel may provide a statistically distinguishable signature of the DNA bases, and may thus allow for rapid DNA sequencing. However, fluctuations of the environment, such as ionic and DNA motion, introduce important scattering processes that may affect the viability of this approach to sequencing. To understand this issue, we have analyzed a simple model that captures the role of this complex environment in electronic dephasing and its ability to remove charge carriers from current carrying states. We find that these effects do not strongly influence the current distributions due to the off-resonant nature of tunneling through the nucleotides -a result we expect to be a common feature of transport in molecular junctions. In particular, only large scattering strengths, as compared to the energetic gap between the molecular states and the Fermi level, significantly alter the form of the current distributions. Since this gap itself is quite large, the current distributions remain protected from this type of noise, further supporting the possibility of using transverse electronic transport measurements for DNA sequencing. PMID- 19804731 TI - Protein-mediated molecular bridging: a key mechanism in biopolymer organization. AB - Protein-mediated bridging is ubiquitous and essential for shaping cellular structures in all organisms. Here we dissect this mechanism for a model system: the Histone-like Nucleoid-Structuring protein (H-NS). We present data from two complementary single-molecule assays that probe the H-NS-DNA interaction: a dynamic optical-trap-driven unzipping assay and an equilibrium H-NS-mediated DNA looping scanning force microscopy imaging assay. To quantitatively analyze and compare these assays, we employ what we consider a novel theoretical framework that describes the bridging motif. The interplay between the experiments and our theoretical model not only infers the effective interaction free energy, the bridging conformation and the duplex-duplex spacing, but also reveals a second, unresolved, cis-binding mode that challenges our current understanding of the role of bridging proteins in chromatin structure. We expect that this theoretical framework for describing protein-mediated bridging will be applicable to proteins acting in chromatin and cytoskeletal organization. PMID- 19804732 TI - Elbow flexibility of the kt38 RNA kink-turn motif investigated by free-energy molecular dynamics simulations. AB - Kink-turns (K-turns) are common structural motifs that can introduce sharp kinks into double-stranded RNA, and have been proposed to mediate large-scale motions in the ribosome. K-turns consist of a bulge loop region flanked by trans sugar Hoogsteen G:A pairs, and the sharp kink conformation is stabilized by A-minor interactions (adenine contacting a G:C basepair in the minor groove). Umbrella sampling molecular dynamics simulations were used to disrupt an A-minor interaction in the ribosomal kt38 turn and to calculate the associated free energy change. Coupling of umbrella sampling with replica exchanges between neighboring umbrella-sampling intervals could further improve the convergence of the free-energy calculations. The simulations revealed a coupled A-minor disruption and global opening of the K-turn motif, and allowed us to characterize several intermediate A-minor conformations. The calculated free-energy profile indicated a meta-stable, semi-open structure of slightly higher free energy ( approximately 1 kcal mol(-1)), separated by a small free-energy barrier ( approximately 1.5 kcal mol(-1)) from the closed (highly kinked) form. Both K-turn states are stabilized by distinct variants of the A-minor interaction. Further opening of the K-turn structure required significantly larger free-energy changes. The semi-open form had a reduced kink angle compatible with experimental data on K-turn solution structures, and opening was coupled to a continuous global unwinding of the K-turn motif. The range of free-energy changes associated with kt38 opening and unwinding are compatible with the idea that K-turns may facilitate biologically relevant motions during large-scale ribosome dynamics. PMID- 19804733 TI - Small-molecule selectively recognizes human telomeric G-quadruplex DNA and regulates its conformational switch. AB - Structural complexity is an inherent feature of the human telomeric sequence, and it presents a major challenge for developing ligands of pharmaceutical interest. Recent studies have pointed out that the induction of a quadruplex or change of a quadruplex conformation on binding may be the most powerful method to exert the desired biological effect. In this study, we demonstrate a quadruplex ligand that binds selectively to different forms of the human telomeric G-quadruplex structure and regulates its conformational switch. The results show that not only can oxazine750 selectively induce parallel quadruplex formation from a random coil telomeric oligonucleotide in the absence of added cations, it also can easily surpass the energy barrier between two structures and change the G quadruplex conformation in Na(+) or K(+) solution. The combination of its unique properties, including the size and shape of the G-quadruplex and the small molecule, is proposed as the predominant force for regulating the special structural formation and transitions. These results may stimulate the design of new quadruplex binders that would be capable of discriminating different G quadruplex structures as well as controlling biological phenomena, functional molecules, and nanomaterials. PMID- 19804734 TI - The semiquinone-iron complex of photosystem II: structural insights from ESR and theoretical simulation; evidence that the native ligand to the non-heme iron is carbonate. AB - The semiquinone-iron complex of photosystem II was studied using electron spin resonance (ESR) spectroscopy and density functional theory calculations. Two forms of the signal were investigated: 1), the native g approximately 1.9 form; and 2), the g approximately 1.84 form, which is well known in purple bacterial reaction centers and occurs in photosystem II when treated with formate. The g approximately 1.9 form shows low- and high-field edges at g approximately 3.5 and g < 0.8, respectively, and resembles the g approximately 1.84 form in terms of shape and width. Both types of ESR signal were simulated using the theoretical approach used previously for the BRC complex, a spin Hamiltonian formalism in which the semiquinone radical magnetically interacts (J approximately 1 cm(-1)) with the nearby high-spin Fe(2+). The two forms of ESR signal differ mainly by an axis rotation of the exchange coupling tensor (J) relative to the zero-field tensor (D) and a small increase in the zero-field parameter D ( approximately 6 cm(-1)). Density functional theory calculations were conducted on model semiquinone-iron systems to identify the physical nature of these changes. The replacement of formate (or glutamate in the bacterial reaction centers) by bicarbonate did not result in changes in the coupling environment. However, when carbonate (CO(3)(2-)) was used instead of bicarbonate, the exchange and zero field tensors did show changes that matched those obtained from the spectral simulations. This indicates that 1), the doubly charged carbonate ion is responsible for the g approximately 1.9 form of the semiquinone-iron signal; and 2), carbonate, rather than bicarbonate, is the ligand to the iron. PMID- 19804735 TI - Unusual thermal disassembly of the SPFH domain oligomer from Pyrococcus horikoshii. AB - Stomatin, prohibitin, flotillin, and HflK/C (SPFH) domain proteins are membrane proteins that are widely conserved from bacteria to mammals. The molecular functions of these proteins have not been established. In mammals, the domain is often found in raft-associated proteins such as flotillin and podocin. We determined the structure of the SPFH domain of PH0470 derived from Pyrococcus horikoshii using NMR. The structure closely resembles that of the SPFH domain of the paralog PH1511, except for two C-terminal helices. The results show that the SPFH domain forms stable dimers, trimers, tetramers, and multimers, although it lacks the coiled-coil region for oligomerization, which is a highly conserved region in this protein family. The oligomers exhibited unusual thermodynamic behavior, as determined by circular dichroism, NMR, gel filtration, chemical cross-linking, and analytical ultracentrifugation. The oligomers were converted into monomers when they were heated once and then cooled. This transition was one way and irreversible. We propose a mechanism of domain swapping for forming dimers as well as successive oligomers. The results of this study provide what to our knowledge are new insights into the common molecular function of the SPFH domain, which may act as a membrane skeleton through oligomerization by domain swapping. PMID- 19804736 TI - Vortex-induced injectable silk fibroin hydrogels. AB - A novel, to our knowledge, technique was developed to control the rate of beta sheet formation and resulting hydrogelation kinetics of aqueous, native silk solutions. Circular dichroism spectroscopy indicated that vortexing aqueous solutions of silkworm silk lead to a transition from an overall protein structure that is initially rich in random coil to one that is rich in beta-sheet content. Dynamic oscillatory rheology experiments collected under the same assembly conditions as the circular dichroism experiments indicated that the increase in beta-sheet content due to intramolecular conformational changes and intermolecular self-assembly of the silk fibroin was directly correlated with the subsequent changes in viscoelastic properties due to hydrogelation. Vortexing low viscosity silk solutions lead to orders-of-magnitude increase in the complex shear modulus, G*, and formation of rigid hydrogels (G* approximately 70 kPa for 5.2 wt % protein concentration). Vortex-induced, beta-sheet-rich silk hydrogels consisted of permanent, physical, intermolecular crosslinks. The hydrogelation kinetics could be controlled easily (from minutes to hours) by changing the vortex time, assembly temperature and/or protein concentration, providing a useful timeframe for cell encapsulation. The stiffness of preformed hydrogels recovered quickly, immediately after injection through a needle, enabling the potential use of these systems for injectable cell delivery scaffolds. PMID- 19804737 TI - Elastic moduli of collagen gels can be predicted from two-dimensional confocal microscopy. AB - We quantitatively compare data obtained from imaging two-dimensional slices of three-dimensional unlabeled and fluorescently labeled collagen gels with confocal reflectance microscopy (CRM) and/or confocal fluorescence microscopy (CFM). Different network structures are obtained by assembling the gels over a range of concentrations at various temperatures. Comparison between CRM and CFM shows that the techniques are not equally sensitive to details of network structure, with CFM displaying higher fidelity in imaging fibers parallel to the optical axis. Comparison of CRM of plain and labeled collagen gels shows that labeling itself induces changes in gel structure, chiefly through inhibition of fibril bundling. Despite these differences, image analyses carried out on two-dimensional CFM and CRM slices of collagen gels reveal identical trends in structural parameters as a function of collagen concentration and gelation temperature. Fibril diameter approximated from either CRM or CFM is in good accord with that determined via electron microscopy. Two-dimensional CRM images are used to show that semiflexible polymer theory can relate network structural properties to elastic modulus successfully. For networks containing bundled fibrils, it is shown that average structural diameter, rather than fibril diameter, is the length scale that sets the magnitude of the gel elastic modulus. PMID- 19804738 TI - Elucidating the mechanism behind irreversible deformation of viral capsids. AB - Atomic force microscopy has recently provided highly precise measurements of mechanical properties of various viruses. However, molecular details underlying viral mechanics remain unresolved. Here we report atomic force microscopy nanoindentation experiments on T=4 hepatitis B virus (HBV) capsids combined with coarse-grained molecular dynamics simulations, which permit interpretation of experimental results at the molecular level. The force response of the indented capsid recorded in simulations agrees with experimental observations. In both experiment and simulation, irreversible capsid deformation is observed for deep indentations. Simulations show the irreversibility to be due to local bending and shifting of capsid proteins, rather than their global rearrangement. These results emphasize the viability of large capsid deformations without significant changes of the mutual positions of HBV capsid proteins, in contrast to the stiffer capsids of other viruses, which exhibit more extensive contacts between their capsid proteins than seen in the case of HBV. PMID- 19804740 TI - Nanosecond motions in proteins impose bounds on the timescale distributions of local dynamics. AB - We elucidate the physics of protein dynamical transition via 10-100-ns molecular dynamics simulations at temperatures spanning 160-300 K. By tracking the energy fluctuations, we show that the protein dynamical transition is marked by a crossover from nonstationary to stationary processes that underlie the dynamics of protein motions. A two-timescale function captures the nonexponential character of backbone structural relaxations. One timescale is attributed to the collective segmental motions and the other to local relaxations. The former is well defined by a single-exponential, nanosecond decay, operative at all temperatures. The latter is described by a set of processes that display a distribution of timescales. Although their average remains on the picosecond timescale, the distribution is markedly contracted at the onset of the transition. It is shown that the collective motions impose bounds on timescales spanned by local dynamical processes. The nonstationary character below the transition implicates the presence of a collection of substates whose interactions are restricted. At these temperatures, a wide distribution of local motion timescales, extending beyond that of nanoseconds, is observed. At physiological temperatures, local motions are confined to timescales faster than nanoseconds. This relatively narrow window makes possible the appearance of multiple channels for the backbone dynamics to operate. PMID- 19804739 TI - Molecular dynamics simulations of Ibuprofen binding to Abeta peptides. AB - Using replica exchange molecular dynamics simulations and the implicit solvent model we probed binding of ibuprofen to Abeta(10-40) monomers and amyloid fibrils. We found that the concave (CV) fibril edge has significantly higher binding affinity for ibuprofen than the convex edge. Furthermore, binding of ibuprofen to Abeta monomers, as compared to fibrils, results in a smaller free energy gain. The difference in binding free energies is likely to be related to the presence of the groove on the CV fibril edge, in which ibuprofen tends to accumulate. The confinement effect of the groove promotes the formation of large low-energy ibuprofen clusters, which rarely occur on the surface of Abeta monomers. These observations led us to suggest that the ibuprofen binding mechanism for Abeta fibrils is different from that for monomers. In general, ibuprofen shows a preference to bind to those regions of Abeta monomers (amino terminal) and fibrils (the CV edge) that are also the primary aggregation interfaces. Based on our findings and on available experimental data, we propose a rationale for the ibuprofen antiaggregation effect. PMID- 19804741 TI - Specific membrane binding of neurotoxin II can facilitate its delivery to acetylcholine receptor. AB - The action of three-finger snake alpha-neurotoxins at their targets, nicotinic acetylcholine receptors (nAChR), is widely studied because of its biological and pharmacological relevance. Most such studies deal only with ligands and receptor models; however, for many ligand/receptor systems the membrane environment may affect ligand binding. In this work we focused on binding of short-chain alpha neurotoxin II (NTII) from Naja oxiana to the native-like lipid bilayer, and the possible role played by the membrane in delivering the toxin to nAChR. Experimental (NMR and mutagenesis) and molecular modeling (molecular-dynamics simulation) studies revealed a specific interaction of the toxin molecule with the phosphatidylserine headgroup of lipids, resulting in the proper topology of NTII on lipid bilayers favoring the attack of nAChR. Analysis of short-chain alpha-neurotoxins showed that most of them possess a high positive charge and sequence homology in the lipid-binding motif of NTII, implying that interaction with the membrane surrounding nAChR may be common for the toxin family. PMID- 19804742 TI - Coherent movement of cell layers during wound healing by image correlation spectroscopy. AB - We have determined the complex sequence of events from the point of injury until reepithelialization in axolotl skin explant model and shown that cell layers move coherently driven by cell swelling after injury. We quantified three-dimensional cell migration using correlation spectroscopy and resolved complex dynamics such as the formation of dislocation points and concerted cell motion. We quantified relative behavior such as velocities and swelling of cells as a function of cell layer during healing. We propose that increased cell volume ( approximately 37% at the basal layer) is the driving impetus for the start of cell migration after injury where the enlarged cells produce a point of dislocation that foreshadows and dictates the initial direction of the migrating cells. Globally, the cells follow a concerted vortex motion that is maintained after wound closure. Our results suggest that cell volume changes the migration of the cells after injury. PMID- 19804744 TI - Taming supplemental material. PMID- 19804743 TI - Diagnosis of cell death by means of infrared spectroscopy. AB - Fourier transform infrared (FTIR) spectroscopy has been established as a fast spectroscopic method for biochemical analysis of cells and tissues. In this research we aimed to investigate FTIR's utility for identifying and characterizing different modes of cell death, using leukemic cell lines as a model system. CCRF-CEM and U937 leukemia cells were treated with arabinoside and doxorubicin apoptosis inducers, as well as with potassium cyanide, saponin, freezing-thawing, and H(2)O(2) necrosis inducers. Cell death mode was determined by various gold standard biochemical methods in parallel with FTIR-microscope measurements. Both cell death modes exhibit large spectral changes in lipid absorbance during apoptosis and necrosis; however, these changes are similar and thus cannot be used to distinguish apoptosis from necrosis. In contrast to the above confounding factor, our results reveal that apoptosis and necrosis can still be distinguished by the degree of DNA opaqueness to infrared light. Moreover, these two cell death modes also can be differentiated by their infrared absorbance, which relates to the secondary structure of total cellular protein. In light of these findings, we conclude that, because of its capacity to monitor multiple biomolecular parameters, FTIR spectroscopy enables unambiguous and easy analysis of cell death modes and may be useful for biochemical and medical applications. PMID- 19804745 TI - Taking risks to transform science. AB - Tackling long-standing problems in science or refuting dogma is not necessarily the best strategy for a young investigator seeking tenure. But just like investing in risky stocks when young, perhaps it should be. Amy Maxmen reports. PMID- 19804746 TI - Learning how to read ADP-ribosylation. AB - ADP-ribosylation is a posttranslational modification that is emerging as a broadly used mechanism to regulate the functions of proteins and their interactions. Recent findings by three groups (Ahel et al., 2009; Gottschalk et al., 2009; Timinszky et al., 2009) establish that proteins with macrodomains bind poly-ADP-ribose to mediate the cellular response to DNA damage. PMID- 19804747 TI - Sex on the brain. AB - Release of testosterone during early development is necessary for masculinization of brain structures in rodents. Wu et al. (2009) now elucidate the neuronal changes caused by aromatase-mediated conversion of testosterone into estrogen in perinatal mice and the ensuing effects on adult behavior. PMID- 19804748 TI - Condensin(g) crossover control to a few breaks. AB - Meiotic chromosome pairs must receive at least one crossover to ensure proper segregation at the first meiotic division. Mets and Meyer (2009) now present compelling evidence that the establishment of higher-order chromosome structure by a condensin complex regulates crossover recombination by controlling the distribution and frequency of meiotic double-strand breaks. PMID- 19804749 TI - It's all about timing. AB - In the formation of long-term memories, a "spaced" distribution of study sessions is more beneficial than closely spaced "massed" study sessions. Pagani et al. (2009) examine the molecular basis of this spacing effect in Drosophila and find a role for the SHP2 homolog, corkscrew, an activator of Ras/MAPK signaling, in establishing optimal spacing intervals. PMID- 19804750 TI - DNA double-strand breaks come into focus. AB - The Mre11-Rad50-Nbs1 (MRN) complex senses DNA double-strand breaks and recruits different repair pathway and checkpoint proteins to break foci. Two new studies (Williams et al., 2009; Lloyd et al., 2009) identify Nbs1 as a key factor in this process and reveal how an N-terminal protein recruitment module in Nbs1 binds to different response factors through shared phosphopeptide motifs. PMID- 19804751 TI - Regulating the regulators: posttranslational modifications of RNA silencing factors. AB - Every regulator should be regulated, and this holds true for small RNAs and their associated proteins. Knowledge has begun to emerge of the various mechanisms that impose specificity on the expression and function of RNA silencing factors. Recent papers, including one in this issue of Cell (Paroo et al., 2009), now reveal the posttranslational modifications that take part in the regulation of the core RNA silencing factors, Ago, Piwi, and TRBP. PMID- 19804752 TI - Mechanotransduction by hair cells: models, molecules, and mechanisms. AB - Mechanotransduction, the transformation of mechanical force into an electrical signal, allows living organisms to hear, register movement and gravity, detect touch, and sense changes in cell volume and shape. Hair cells in the inner ear are specialized mechanoreceptor cells that detect sound and head movement. The mechanotransduction machinery of hair cells is extraordinarily sensitive and responds to minute physical displacements on a submillisecond timescale. The recent discovery of several molecular constituents of the mechanotransduction machinery of hair cells provides a new framework for the interpretation of biophysical data and necessitates revision of prevailing models of mechanotransduction. PMID- 19804754 TI - Estrogen masculinizes neural pathways and sex-specific behaviors. AB - Sex hormones are essential for neural circuit development and sex-specific behaviors. Male behaviors require both testosterone and estrogen, but it is unclear how the two hormonal pathways intersect. Circulating testosterone activates the androgen receptor (AR) and is also converted into estrogen in the brain via aromatase. We demonstrate extensive sexual dimorphism in the number and projections of aromatase-expressing neurons. The masculinization of these cells is independent of AR but can be induced in females by either testosterone or estrogen, indicating a role for aromatase in sexual differentiation of these neurons. We provide evidence suggesting that aromatase is also important in activating male-specific aggression and urine marking because these behaviors can be elicited by testosterone in males mutant for AR and in females subjected to neonatal estrogen exposure. Our results suggest that aromatization of testosterone into estrogen is important for the development and activation of neural circuits that control male territorial behaviors. PMID- 19804753 TI - Olfactory perception: receptors, cells, and circuits. AB - Remarkable advances in our understanding of olfactory perception have been made in recent years, including the discovery of new mechanisms of olfactory signaling and new principles of olfactory processing. Here, we discuss the insight that has been gained into the receptors, cells, and circuits that underlie the sense of smell. PMID- 19804755 TI - Nbs1 flexibly tethers Ctp1 and Mre11-Rad50 to coordinate DNA double-strand break processing and repair. AB - The Nijmegen breakage syndrome 1 (Nbs1) subunit of the Mre11-Rad50-Nbs1 (MRN) complex protects genome integrity by coordinating double-strand break (DSB) repair and checkpoint signaling through undefined interactions with ATM, MDC1, and Sae2/Ctp1/CtIP. Here, fission yeast and human Nbs1 structures defined by X ray crystallography and small angle X-ray scattering (SAXS) reveal Nbs1 cardinal features: fused, extended, FHA-BRCT(1)-BRCT(2) domains flexibly linked to C terminal Mre11- and ATM-binding motifs. Genetic, biochemical, and structural analyses of an Nbs1-Ctp1 complex show Nbs1 recruits phosphorylated Ctp1 to DSBs via binding of the Nbs1 FHA domain to a Ctp1 pThr-Asp motif. Nbs1 structures further identify an extensive FHA-BRCT interface, a bipartite MDC1-binding scaffold, an extended conformational switch, and the molecular consequences associated with cancer predisposing Nijmegen breakage syndrome mutations. Tethering of Ctp1 to a flexible Nbs1 arm suggests a mechanism for restricting DNA end processing and homologous recombination activities of Sae2/Ctp1/CtIP to the immediate vicinity of DSBs. PMID- 19804756 TI - A supramodular FHA/BRCT-repeat architecture mediates Nbs1 adaptor function in response to DNA damage. AB - The Mre11/Rad50/Nbs1 protein complex plays central enzymatic and signaling roles in the DNA-damage response. Nuclease (Mre11) and scaffolding (Rad50) components of MRN have been extensively characterized, but the molecular basis of Nbs1 function has remained elusive. Here, we present a 2.3A crystal structure of the N terminal region of fission yeast Nbs1, revealing an unusual but conserved architecture in which the FHA- and BRCT-repeat domains structurally coalesce. We demonstrate that diphosphorylated pSer-Asp-pThr-Asp motifs, recently identified as multicopy docking sites within Mdc1, are evolutionarily conserved Nbs1 binding targets. Furthermore, we show that similar phosphomotifs within Ctp1, the fission yeast ortholog of human CtIP, promote interactions with the Nbs1 FHA domain that are necessary for Ctp1-dependent resistance to DNA damage. Finally, we establish that human Nbs1 interactions with Mdc1 occur through both its FHA- and BRCT repeat domains, suggesting how their structural and functional interdependence underpins Nbs1 adaptor functions in the DNA-damage response. PMID- 19804757 TI - Phosphorylation of the human microRNA-generating complex mediates MAPK/Erk signaling. AB - MicroRNAs (miRNAs) govern an expanding number of biological and disease processes. Understanding the mechanisms by which the miRNA pathway is regulated, therefore, represents an important area of investigation. We determined that the human miRNA-generating complex is comprised of Dicer and phospho-TRBP isoforms. Phosphorylation of TRBP is mediated by the mitogen-activated protein kinase (MAPK) Erk. Expression of phospho-mimic TRBP and TRBP phosphorylation enhanced miRNA production by increasing stability of the miRNA-generating complex. Mitogenic signaling in response to serum and the tumor promoter PMA was dependent on TRBP phosphorylation. These effects were accompanied by a coordinated increase in levels of growth-promoting miRNA and reduced expression of let-7 tumor suppressor miRNA. Conversely, pharmacological inhibition of MAPK/Erk resulted in an anti-growth miRNA profile. Taken together, these studies indicate that the MAPK/Erk pathway regulates the miRNA machinery and suggest a general principle, wherein signaling systems target the miRNA pathway to achieve biological responses. PMID- 19804758 TI - The Argonaute CSR-1 and its 22G-RNA cofactors are required for holocentric chromosome segregation. AB - RNAi-related pathways regulate diverse processes, from developmental timing to transposon silencing. Here, we show that in C. elegans the Argonaute CSR-1, the RNA-dependent RNA polymerase EGO-1, the Dicer-related helicase DRH-3, and the Tudor-domain protein EKL-1 localize to chromosomes and are required for proper chromosome segregation. In the absence of these factors chromosomes fail to align at the metaphase plate and kinetochores do not orient to opposing spindle poles. Surprisingly, the CSR-1-interacting small RNAs (22G-RNAs) are antisense to thousands of germline-expressed protein-coding genes. Nematodes assemble holocentric chromosomes in which continuous kinetochores must span the expressed domains of the genome. We show that CSR-1 interacts with chromatin at target loci but does not downregulate target mRNA or protein levels. Instead, our findings support a model in which CSR-1 complexes target protein-coding domains to promote their proper organization within the holocentric chromosomes of C. elegans. PMID- 19804759 TI - CDE-1 affects chromosome segregation through uridylation of CSR-1-bound siRNAs. AB - We have studied the function of a conserved germline-specific nucleotidyltransferase protein, CDE-1, in RNAi and chromosome segregation in C. elegans. CDE-1 localizes specifically to mitotic chromosomes in embryos. This localization requires the RdRP EGO-1, which physically interacts with CDE-1, and the Argonaute protein CSR-1. We found that CDE-1 is required for the uridylation of CSR-1 bound siRNAs, and that in the absence of CDE-1 these siRNAs accumulate to inappropriate levels, accompanied by defects in both meiotic and mitotic chromosome segregation. Elevated siRNA levels are associated with erroneous gene silencing, most likely through the inappropriate loading of CSR-1 siRNAs into other Argonaute proteins. We propose a model in which CDE-1 restricts specific EGO-1-generated siRNAs to the CSR-1 mediated, chromosome associated RNAi pathway, thus separating it from other endogenous RNAi pathways. The conserved nature of CDE-1 suggests that similar sorting mechanisms may operate in other animals, including mammals. PMID- 19804760 TI - 4E-BP extends lifespan upon dietary restriction by enhancing mitochondrial activity in Drosophila. AB - Dietary restriction (DR) extends lifespan in multiple species. To examine the mechanisms of lifespan extension upon DR, we assayed genome-wide translational changes in Drosophila. A number of nuclear encoded mitochondrial genes, including those in Complex I and IV of the electron transport chain, showed increased ribosomal loading and enhanced overall activity upon DR. We found that various mitochondrial genes possessed shorter and less structured 5'UTRs, which were important for their enhanced mRNA translation. The translational repressor 4E-BP, the eukaryotic translation initiation factor 4E binding protein, was upregulated upon DR and mediated DR dependent changes in mitochondrial activity and lifespan extension. Inhibition of individual mitochondrial subunits from Complex I and IV diminished the lifespan extension obtained upon DR, reflecting the importance of enhanced mitochondrial function during DR. Our results imply that translational regulation of nuclear-encoded mitochondrial gene expression by 4E-BP plays an important role in lifespan extension upon DR. For a video summary of this article, see the PaperFlick file with the Supplemental Data available online. PMID- 19804761 TI - Stringent specificity in the construction of a GABAergic presynaptic inhibitory circuit. AB - GABAergic interneurons are key elements in neural coding, but the mechanisms that assemble inhibitory circuits remain unclear. In the spinal cord, the transfer of sensory signals to motor neurons is filtered by GABAergic interneurons that act presynaptically to inhibit sensory transmitter release and postsynaptically to inhibit motor neuron excitability. We show here that the connectivity and synaptic differentiation of GABAergic interneurons that mediate presynaptic inhibition is directed by their sensory targets. In the absence of sensory terminals these GABAergic neurons shun other available targets, fail to undergo presynaptic differentiation, and withdraw axons from the ventral spinal cord. A sensory-specific source of brain derived neurotrophic factor induces synaptic expression of the GABA synthetic enzyme GAD65--a defining biochemical feature of this set of interneurons. The organization of a GABAergic circuit that mediates presynaptic inhibition in the mammalian CNS is therefore controlled by a stringent program of sensory recognition and signaling. PMID- 19804762 TI - Retinal input instructs alignment of visual topographic maps. AB - Sensory information is represented in the brain in the form of topographic maps, in which neighboring neurons respond to adjacent external stimuli. In the visual system, the superior colliculus receives topographic projections from the retina and primary visual cortex (V1) that are aligned. Alignment may be achieved through the use of a gradient of shared axon guidance molecules, or through a retinal-matching mechanism in which axons that monitor identical regions of visual space align. To distinguish between these possibilities, we take advantage of genetically engineered mice that we show have a duplicated functional retinocollicular map but only a single map in V1. Anatomical tracing revealed that the corticocollicular projection bifurcates to align with the duplicated retinocollicular map in a manner dependent on the normal pattern of spontaneous activity during development. These data suggest a general model in which convergent maps use coincident activity patterns to achieve alignment. PMID- 19804763 TI - The phosphatase SHP2 regulates the spacing effect for long-term memory induction. AB - A property of long-term memory (LTM) induction is the requirement for repeated training sessions spaced over time. This augmentation of memory formation with spaced resting intervals is called the spacing effect. We now show that in Drosophila, the duration of resting intervals required for inducing LTM is regulated by activity levels of the protein tyrosine phosphatase corkscrew (CSW). Overexpression of wild-type CSW in mushroom body neurons shortens the inter-trial interval required for LTM induction, whereas overexpression of constitutively active CSW proteins prolongs these resting intervals. These gain-of-function csw mutations are associated with a clinical condition of mental retardation. Biochemical analysis reveals that LTM-inducing training regimens generate repetitive waves of CSW-dependent MAPK activation, the length of which appears to define the duration of the resting interval. Constitutively active CSW proteins prolong the resting interval by altering the MAPK inactivation cycle. We thus provide insight into the molecular basis of the spacing effect. PMID- 19804764 TI - Enhanced SnapShot: Antibiotic inhibition of protein synthesis II. AB - The translational apparatus is one of the major targets for antibiotics in the bacterial cell. Antibiotics predominantly interact with the functional centers of the ribosome, namely the messenger RNA (mRNA)-transfer RNA (tRNA) decoding region on the 30S subunit, the peptidyltransferase center on the 50S subunit, or the ribosomal exit tunnel through which the nascent polypeptide chain passes during translation. Protein synthesis can be divided into three phases: initiation, elongation, and termination/recycling. PMID- 19804765 TI - Vertical grid test and modified horizontal grid test are sensitive methods for evaluating motor dysfunctions in the MPTP mouse model of Parkinson's disease. AB - Parkinson's disease (PD) is caused by selective degeneration of the nigral dopaminergic (DArgic) neurons and is accompanied by motor dysfunctions such as tremor, akinesia, and rigidity. Changes in the degree of motor deficit can be utilized as a noninvasive way of assessing alterations in the number of DArgic neurons and/or the amount of DA in animal models of PD, such as mice systemically administrated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). In this study, in order to develop sensitive methods to detect DA-associated motor deficits, we designed a new test called vertical grid test and modified the existing horizontal grid test. After acute MPTP treatment, decreases in the levels of striatal DA (17.4% of control), dihydroxyphenylacetic acid (33.3%), and homovanillic acid (40.5%) were observed. On the modified horizontal grid test, the MPTP-administered mice exhibited average forelimb step distance that was lower than control (82.58%) and correlated with the striatal DA levels. On the vertical grid test, the MPTP-treated mice took dramatically longer total time to climb down (220.94%) and time to make the turn (339.29%) compared to control, and this correlated well with the degree of striatal DA depletion. In comparison, the gait test produced only a small, albeit statistically significant, reduction in the mean stride length (94.55% of control). These results show that the vertical grid test can provide a sensitive measure of motor deficit in mice following administration of MPTP. PMID- 19804766 TI - Hypertonic saline attenuates tissue loss and astrocyte hypertrophy in a model of traumatic brain injury. AB - Hypertonic saline is currently being used in the treatment of patients with post traumatic cerebral edema and elevated intracranial pressure resulting from TBI. A limited number of studies show the cellular effects of hypertonic saline and no studies, to our knowledge, have investigated the effects on astrocytes. The role of astrocyte responses after traumatic brain injury remains unclear. There is evidence that reduced astrocyte proliferation is detrimental while increased hypertrophy and proliferation are signs of increased injury severity. Therefore, this study focused on the hypothesis that hypertonic saline-induced improvements in histological outcome are time dependent and may be associated with alterations in astrocyte hypertrophy after cortical contusion injury. Histopathological changes at 7 days after controlled cortical impact (CCI) injury were examined. Brain tissue loss determined using cresyl violet staining and astrocyte hypertrophy and proliferation were assessed using glial fibrillary acidic protein immunostaining in hypertonic saline and normal saline treated rats, and untreated, injured controls. Effects of the timing of hypertonic saline treatment administration on tissue loss were also examined. Plasma osmolarity and sodium levels were measured over 4 h and again at 24 h following hypertonic saline administration. Results show that hypertonic saline treatment reduced tissue loss that correlated with attenuated astrocyte hypertrophy characterized by reductions in astrocyte immunoreactivity without changes in the number of astrocytes after CCI injury. Delayed treatment of hypertonic saline resulted in the greatest reduction in tissue loss compared to all other treatments indicating that there is a therapeutic window for hypertonic saline use after traumatic brain injury. PMID- 19804767 TI - Genetic variant in visfatin gene promoter is associated with decreased risk of coronary artery disease in a Chinese population. AB - BACKGROUND: Visfatin is a newly identified pro-inflammatory adipokine expressed predominantly in visceral fat. Previous studies have suggested a role for visfatin in low-grade inflammation and regulation of lipid metabolism. Most recently, a genetic polymorphism -1535C>T located in the visfatin gene promoter has been identified, and suggested to be associated with the regulation of visfatin expression, lipid levels. However, it is unclear whether this polymorphism has a linkage with CAD. METHODS: We conducted a hospital-based case control study with 257 CAD patients and 292 controls to examine the potential association of the Visfatin -1535C>T polymorphism with CAD. RESULTS: The frequencies of the CC, CT, and TT genotypes in cases were significantly different from those of controls (chi2=6.223, P=0.045). Subjects with the variant genotypes (CT+TT) had a 40% decreased risk of CAD relative to CC carriers (adjusted OR=0.60, 95%CI=0.40-0.89). Furthermore, the adjusted OR of a TT genotype for CAD was 0.52 (95%CI=0.31-0.87). There was a significant association between Visfatin 1535C>T polymorphism and triglyceride levels in both CAD patients and controls (P=0.003, 0.018, respectively). In stratified analyses, the T allele was significantly associated with reduced risk of CAD in males, subjects with age <59years, and non-smokers. Moreover, a borderline statistical significance (P=0.058 for trend) was observed between the variant genotypes and severity of CAD. CONCLUSION: Our results suggested that Visfatin -1535C>T polymorphism might be associated with reduced risk of CAD in a Chinese population. PMID- 19804768 TI - Does acute hyperglycemia add prognostic value to the GRACE score in individuals with non-ST elevation acute coronary syndromes? AB - BACKGROUND: It is not known in what extent admission glucose improves risk stratification of the GRACE Score in patients with non-ST-segment elevation acute coronary syndromes (ACS). We tested the hypothesis that admission glucose adds relevant prognostic information to the GRACE Score. METHODS: Consecutive patients admitted with ACS had plasma glucose measured at admission and cardiovascular events were defined as death, non-fatal myocardial infarction or non-fatal refractory angina during hospitalization. RESULTS: Among the 148 patients studied, 11.5% developed cardiovascular events. Patients in the forth quartile of admission glucose (> or =175mg/dl) had a greater incidence of events, compared with those in the first 3 quartiles (22% vs. 8.1%; RR=2.7; 95%CI 1.1-6.4; P=0.03). Plasma glucose remained a predictor of events, after adjustment for diabetes (P=0.03). After adjustment for the GRACE Score, glucose in the forth quartile lost its predictive value (P=0.29). Plasma glucose added to GRACE did not improve the C-statistics (0.82; 95%CI 0.75-0.88), as compared with the original Score (0.81; 95%CI 0.74-0.87). Net reclassification improvement by new score was -0.03 (P=0.86), indicating no useful reclassification. CONCLUSION: Despite its association with adverse events, admission plasma glucose does not improve GRACE's accuracy to predict in-hospital events in patients with ACS. PMID- 19804769 TI - An anomalous result from gel tubes for vitamin D. PMID- 19804770 TI - Association between recurrent miscarriages and organochlorine pesticide levels. AB - OBJECTIVES: Recurrent miscarriage (RM) is a challenging medical problem because of its unknown pathogenesis and etiology in most of the cases. Recent studies suggest the role of persistent environmental pollutants such as organochlorine pesticides (OCPs) in the etiology of RM. The present study was conducted to investigate possible associations of OCPs in the pathogenesis of RM. DESIGN AND METHODS: Blood OCP levels were analyzed in women with RM (cases) and women with normal full term delivery with live birth (controls) by using a gas chromatograph equipped with an electron capture detector. RESULTS: A statistically significant association (p=0.01) was observed between blood gamma-HCH levels and women with recurrent miscarriages. CONCLUSIONS: This study suggests that high blood levels of gamma-HCH may be associated with risk of RM. PMID- 19804771 TI - Expression of HIF-1 alpha, VEGF and EPO in peripheral blood from patients with two cardiac abnormalities associated with hypoxia. AB - OBJECTIVES: HIF-1 alpha (hypoxia-inducible factor-1 alpha) mediates the responses of mammalian cells to hypoxia/ischemia by inducing the expression of adaptive gene products (e.g., vascular endothelial growth factor (VEGF) and erythropoietin (EPO)). Persistent pulmonary hypertension of the newborn (PPHN) and cyanotic congenital heart disease (CCHD) are common neonatal diseases considered as paradigms of hypoxemia. Since the expression HIF-1 alpha, VEGF and EPO in newborns diagnosed with these diseases has yet to be studied, we set out to define the expression of these genes in peripheral blood from newborn infants diagnosed with PPHN and CCHD. DESIGN AND METHODS: The mRNA transcripts encoding HIF-1 alpha, VEGF and EPO were measured by RT-PCR in healthy newborn infants and infants diagnosed with PPHN and CCHD. RESULTS: An important increase in HIF-1 alpha expression was observed in both pathological conditions, accompanied by significant increases in VEGF and EPO expression when compared to healthy infants. CONCLUSIONS: HIF-1 alpha mRNA expression increases in newborn infants with PPHN or CCHD, as does the expression of its target genes VEGF and EPO. PMID- 19804772 TI - Measurement of cystatin C in capillary blood samples in pediatric patients. AB - We assessed whether the GFR marker cystatin C can be measured reliably in capillary blood samples in children with renal disease. Cystatin C was measured in venous and capillary blood samples obtained simultaneously from 48 children, and GFR was estimated using the Filler equation. Estimated GFR based on capillary cystatin C concentrations was higher than the venous cystatin C based GFR. The limits of agreement between estimated GFR based on capillary and venous measurements exceeded +/-20% at the upper reference concentration, which hampers the clinical usefulness of capillary sampling. PMID- 19804773 TI - Haptoglobin genotypes in Chagas' disease. AB - OBJECTIVES: To investigate the existence of an association between haptoglobin (Hp) genotypes and the severity of heart complications in Chagas' disease. DESIGN AND METHODS: Hp genotyping was performed by PCR in 107 Brazilian patients sub classified in asymptomatic, with mild heart disease and with severe heart disease. RESULTS: Multiple logistic regression (R(2)=24%) indicated that patients with the Hp1-1 genotype have lower probability of developing the severe heart complications. CONCLUSION: The Hp polymorphism may influence the clinical evolution of Chagas' disease. PMID- 19804774 TI - Identification of a mechanochemical checkpoint and negative feedback loop regulating branching morphogenesis. AB - Cleft formation is the initial step in submandibular salivary gland (SMG) branching morphogenesis, and may result from localized actomyosin-mediated cellular contraction. Since ROCK regulates cytoskeletal contraction, we investigated the effects of ROCK inhibition on mouse SMG ex vivo organ cultures. Pharmacological inhibitors of ROCK, isoform-specific ROCK I but not ROCK II siRNAs, as well as inhibitors of myosin II activity stalled clefts at initiation. This finding implies the existence of a mechanochemical checkpoint regulating the transition of initiated clefts into progression-competent clefts. Downstream of the checkpoint, clefts are rendered competent through localized assembly of fibronectin promoted by ROCK I/myosin II. Cleft progression is primarily mediated by ROCK I/myosin II-stimulated cell proliferation with a contribution from cellular contraction. Furthermore, we demonstrate that FN assembly itself promotes epithelial proliferation and cleft progression in a ROCK-dependent manner. ROCK also stimulates a proliferation-independent negative feedback loop to prevent further cleft initiations. These results reveal that cleft initiation and progression are two physically and biochemically distinct processes. PMID- 19804775 TI - A human neuron injury model for molecular studies of axonal regeneration. AB - The enhancement of regeneration of damaged axons in both the peripheral and central nervous systems is a widely pursued goal in clinical medicine. Although some of the molecular mechanisms involved in the intrinsic neurite regeneration program have been elucidated, much additional study is required for development of new therapeutics. The majority of studies in the field of axonal regeneration have utilized animal models due to obvious limitations of the accessibility of human neural tissues. Here we describe the use of human embryonic stem cell (hESC)-derived neurons as a novel model for studying neuronal responses to axonal injury. Neurons were generated using PA6 induction and neurites injured in vitro using trituration or laser microdissection. Lesioned neurons re-extended neurites with distinct growth cones. Expression of proteins associated with regeneration were observed in this human in vitro system, including appearance of importin beta1 in processes after neuritomy. Laser-transected hESC-derived neuronal cultures were analyzed for their transcriptional response to injury using Affymetrix expression microarrays. Profound changes in gene expression were observed over a time course of 2 to 24 hours after lesion. The expression of several genes reported to be involved in axonal injury responses in animal models changed following injury of hESC-derived neurons. Thus, hESC-derived neurons may be a useful in vitro model system for mechanistic studies on human axonal injury and regeneration. PMID- 19804776 TI - Cell sex determines anoikis resistance in vascular smooth muscle cells. AB - Sexual dimorphism, detectable in vascular smooth muscle cells freshly isolated from aorta of male and female rats, is associated with a different susceptibility to radiation-induced apoptosis. In this work we investigated the mechanism underlying this difference and discovered that, in comparison with cells from male rats, cells from female rats show adhesion-associated resistance to apoptosis, the so called anoikis resistance. This is apparently due to a more adhering phenotype, characterized by a well organized actin microfilament cytoskeleton and to an increased phosphorylated focal adhesion kinase, and, more importantly, to a higher propensity to undergo survival by autophagy. PMID- 19804778 TI - Yolk hormones have sex-specific long-term effects on behavior in the pied flycatcher (Ficedula hypoleuca). AB - The hormonal environment during early development, such as maternally derived androgens in bird eggs, shapes the development and phenotype of the offspring in ways that may have important long-term consequences for behaviour. We studied the effects of yolk androgens on multiple behavioural traits in female and male pied flycatchers (Ficedula hypoleuca) by experimentally elevating androgen levels (testosterone and androstenedione) in the eggs. The birds were housed in a common garden environment in captivity until full independence, after which their behaviour was tested. We found that androgen-treated males were more likely than control males to explore a novel environment and showed higher activity in the presence of a novel object. In response to a simulated predator attack, androgen treated males mainly showed freezing behaviour, while control males showed escape behaviour. Females from the androgen treatment and control group showed no differences in these behaviours. Androgen treatment did not affect neophobia (latency to approach the novel object) or dominance behaviour in either sex. Behaviour in the novel environment and towards a novel object was repeatable, but behaviours in the different experiments were mostly not inter-correlated. These results indicate that yolk androgens have various long-lasting effects on behaviour, especially in males, but that they do not induce a distinct behavioural syndrome. As behaviour is strongly linked with fitness, our results suggest that yolk androgens may play a role in determining fitness, and thus play a potentially adaptive role. PMID- 19804777 TI - Two-dimensional pulsed electron spin resonance characterization of 15N-labeled archaeal Rieske-type ferredoxin. AB - Two-dimensional electron spin-echo envelope modulation (ESEEM) analysis of the uniformly (15)N-labeled archaeal Rieske-type [2Fe-2S] ferredoxin (ARF) from Sulfolobus solfataricus P1 has been conducted in comparison with the previously characterized high-potential protein homologs. Major differences among these proteins were found in the hyperfine sublevel correlation (HYSCORE) lineshapes and intensities of the signals in the (++) quadrant, which are contributed from weakly coupled (non-coordinated) peptide nitrogens near the reduced clusters. They are less pronounced in the HYSCORE spectra of ARF than those of the high potential protein homologs, and may account for the tuning of Rieske-type clusters in various redox systems. PMID- 19804779 TI - Seasonality, sociality, and reproduction: Long-term stressors of ring-tailed lemurs (Lemur catta). AB - Fecal glucocorticoid (fGC) concentrations are reliable, non-invasive indices of physiological stress that provide insight into an animal's energetic and social demands. To better characterize the long-term stressors in adult members of a female-dominant, seasonally breeding species - the ring-tailed lemur (Lemur catta) - we first validated fecal samples against serum samples and then examined the relationship between fGC concentrations and seasonal, social, demographic, genetic, and reproductive variables. Between 1999 and 2006, we collected 1386 fecal samples from 32 adult, semi-free-ranging animals of both sexes. In males and non-pregnant, non-lactating females, fGC concentrations were significantly elevated during the breeding season, specifically during periods surrounding known conceptions. Moreover, group composition (e.g., multi-male versus one-male) significantly predicted the fGC concentrations of males and females in all reproductive states. In particular, the social instability introduced by intra male competition likely created a stressor for all animals. We found no relationship, however, between fGC and the sex, age, or heterozygosity of animals. In reproducing females, fGC concentrations were significantly greater during lactation than during the pre-breeding period. During pregnancy, fGC concentrations were elevated in mid-ranking dams, relative to dominant or subordinate dams, and significantly greater during the third trimester than during the first or second trimesters. Thus, in the absence of nutritional stressors, social dominance was a relatively poor predictor of fGC in this female dominant species. Instead, the animals were maximally challenged by their social circumstances and reproductive events-males by competition for mating opportunities and females by late-term gestation and lactation. PMID- 19804780 TI - Increased corticosterone levels in mice subjected to the rat exposure test. AB - In recent years, there has been a notable interest in studying prey-predator relationships to develop rodent-based models for the neurobehavioral aspects of stress and emotion. However, despite the growing use of transgenic mice and results showing important differences in the behavioral responses of rats and mice, little research has been conducted regarding the responses of mice to predators. The rat exposure test (RET), a recently developed and behaviorally validated prey-predator (mouse-rat)-based model, has proven to be a useful tool in evaluating the defensive responses of mice facing rats. To further validate the RET, we investigated the endocrine and behavioral responses of mice exposed to this apparatus. We first constructed a plasma corticosterone secretion curve in mice exposed to a rat or to an empty cage (control). Rat-exposed mice showed a pronounced rise in corticosterone levels that peaked 15 min from the beginning of the predator exposure. The corticosterone levels and behavioral responses of mice exposed to a rat or to a toy in the RET apparatus were then measured. We observed high plasma corticosterone levels along with clear avoidance behaviors represented by decreases in tunnel and surface area exploration and increases in risk assessment behaviors and freezing. This strongly suggests that the test elicits a repertoire of behavioral responses compatible with an aversion state and indicates that it is a promising model for the evaluation of prey-predator interactions. However, more physiological, neurochemical, and pharmacological studies are needed to further validate the test. PMID- 19804781 TI - A novel method for isolating dendritic cells from human bronchoalveolar lavage fluid. AB - BACKGROUND: Dendritic cells (DCs) play a pivotal role in linking the innate and adaptive immune response and have been implicated in a variety of pulmonary diseases. Currently, studies on the role of DCs are limited by difficulties in isolating DCs from the lung. Surgical lung specimens are not readily available and purification of DCs from digested lung tissue is likely to induce phenotypical and functional changes. DCs obtained from the alveolar spaces are thought to represent the local microenvironment and can be obtained using minimally invasive techniques. We developed a novel method of isolating DCs from bronchoalveolar lavage (BAL) fluid. METHODS: After removal of macrophages, the remaining BAL cells were stained with a lineage mix (CD3-, CD14-, CD16-, CD19-, CD56-FITC), CD11c and HLA-DR and sorted with a FACS ARIA. DAPI was used as a dead live marker. mDCs were low autofluorescent, lineage mix negative, CD11c+ and HLA DR+ cells. pDCs were CD11c(-) but CD123+. Morphological assessment of sorted mDCs and pDCs was performed. Sorted mDCs were tested in a mixed leukocyte reaction (MLR) with naive CD4+ T cells and evaluated for T cell differentiation and cytokine production. With confocal microscopy DC-T cell interaction was assessed. RESULTS: Using our sorting strategy, mDCs and pDCs, with a high purity upon FACS analysis of the sorted fraction, were obtained. These cells showed the morphological characteristics of DCs. Most importantly, mDCs were able to induce T cell proliferation and differentiation in a MLR, and interact with T cells as assessed by confocal microscopy. These results indicate the presence of functional DCs. Freezing and thawing of the BAL cells did not affect phenotype or T cell stimulatory capacity of the isolated DCs. CONCLUSION: Using a novel sorting strategy, functional mDCs can be isolated from BAL fluid, enabling a detailed study in pulmonary disease. PMID- 19804782 TI - Mapping the structure of an integral membrane protein under semi-denaturing conditions by laser-induced oxidative labeling and mass spectrometry. AB - Little is known about the structural properties of semi-denatured membrane proteins. The current study employs laser-induced oxidative labeling of methionine side chains in combination with electrospray mass spectrometry and optical spectroscopy for gaining insights into the conformation of bacteriorhodopsin (BR) under partially denaturing conditions. The native protein shows extensive oxidation at M32, M68, and M163, which are located in solvent accessible loops. In contrast, M20 (helix A), M56/60 (helix B), M118 (helix D), M145 (helix E), and M209 (helix G) are strongly protected, consistent with the known protein structure. Exposure of the protein to acidic conditions leads to a labeling pattern very similar to that of the native state. The absence of large scale conformational changes at low pH is in agreement with recent crystallography data. Solubilization of BR in SDS induces loss of the retinal chromophore concomitant with collapse of the binding pocket, thereby precluding solvent access to the protein interior. Tryptophan fluorescence data confirm the presence of a large protein core that remains protected from water. However, oxidative labeling indicates partial unfolding of helices A and D in SDS. Irreversible thermal denaturation of the protein at 100 degrees C induces a labeling pattern quite similar to that seen upon SDS exposure. Labeling experiments on refolded bacterioopsin reveal a native-like structure, but with partial unfolding of helix D. Our data suggest that noncovalent contacts with the retinal chromophore in native BR play an important role for the stability of this particular helix. Overall, the present work illustrates the viability of using laser-induced oxidative labeling as a novel tool for characterizing structural changes of membrane proteins in response to alterations of their solvent environment. PMID- 19804784 TI - A lower-order oligomer form of phage shock protein A (PspA) stably associates with the hexameric AAA(+) transcription activator protein PspF for negative regulation. AB - To survive and colonise their various environments, including those used during infection, bacteria have developed a variety of adaptive systems. Amongst these is phage shock protein (Psp) response, which can be induced in Escherichia coli upon filamentous phage infection (specifically phage secretin pIV) and by other membrane-damaging agents. The E. coli Psp system comprises seven proteins, of which PspA is the central component. PspA is a bifunctional protein that is directly involved in (i) the negative regulation of the psp-specific transcriptional activator PspF and (ii) the maintenance of membrane integrity in a mechanism proposed to involve the formation of a 36-mer ring complex. Here we established that the PspA negative regulation of PspF ATPase activity is the result of a cooperative inhibition. We present biochemical evidence showing that an inhibitory PspA-PspF regulatory complex, which has significantly reduced PspF ATPase activity, is composed of around six PspF subunits and six PspA subunits, suggesting that PspA exists in at least two different oligomeric assemblies. We now establish that all four putative helical domains of PspA are critical for the formation of the 36-mer. In contrast, not all four helical domains are required for the formation of the inhibitory PspA-PspF complex. Since a range of initial PspF oligomeric states permit formation of the apparent PspA-PspF dodecameric assembly, we conclude that PspA and PspF demonstrate a strong propensity to self assemble into a single defined heteromeric regulatory complex. PMID- 19804783 TI - The structure of the N-terminus of kindlin-1: a domain important for alphaiibbeta3 integrin activation. AB - The integrin family of heterodimeric cell adhesion molecules exists in both low- and high-affinity states, and integrin activation requires binding of the talin FERM (four-point-one, ezrin, radixin, moesin) domain to membrane-proximal sequences in the beta-integrin cytoplasmic domain. However, it has recently become apparent that the kindlin family of FERM domain proteins is also essential for talin-induced integrin activation. FERM domains are typically composed of F1, F2, and F3 domains, but the talin FERM domain is atypical in that it contains a large insert in F1 and is preceded by a previously unrecognized domain, F0. Initial sequence alignments showed that the kindlin FERM domain was most similar to the talin FERM domain, but the homology appeared to be restricted to the F2 and F3 domains. Based on a detailed characterization of the talin FERM domain, we have reinvestigated the sequence relationship with kindlins and now show that kindlins do indeed contain the same domain structure as the talin FERM domain. However, the kindlin F1 domain contains an even larger insert than that in talin F1 that disrupts the sequence alignment. The insert, which varies in length between different kindlins, is not conserved and, as in talin, is largely unstructured. We have determined the structure of the kindlin-1 F0 domain by NMR, which shows that it adopts the same ubiquitin-like fold as the talin F0 and F1 domains. Comparison of the kindlin-1 and talin F0 domains identifies the probable interface with the kindlin-1 F1 domain. Potential sites of interaction of kindlin F0 with other proteins are discussed, including sites that differ between kindlin 1, kindlin-2, and kindlin-3. We also demonstrate that F0 is required for the ability of kindlin-1 to support talin-induced alphaIIbbeta3 integrin activation and for the localization of kindlin-1 to focal adhesions. PMID- 19804785 TI - Neuroprotective role of curcumin in the cerebellum of streptozotocin-induced diabetic rats. AB - AIMS: Chronic hyperglycaemia in diabetes involves a direct neuronal damage caused by intracellular glucose which leads to altered neurotransmitter functions and reduced motor activity. The present study investigated the effect of curcumin in the functional regulation of muscarinic and alpha7 nicotinic acetylcholine receptors, insulin receptors, acetylcholine esterase and Glut3 in the cerebellum of streptozotocin (STZ)-induced diabetic rats. MAIN METHODS: All studies were done in the cerebellum of male Wistar rats. Radioreceptor binding assays were done for total muscarinic, M(1) and M(3) receptors using specific ligands, and the gene expression was also studied using specific probes. KEY FINDINGS: Our results showed an increased gene expression of acetylcholine esterase, Glut3, muscarinic M1, M3, alpha7 nicotinic acetylcholine and insulin receptors in the cerebellum of diabetic rats in comparison to control. Scatchard analysis of total muscarinic, M1 and M3 receptors showed an increased binding parameter, B(max) in diabetic rats compared to control. Curcumin and insulin inhibited diabetes induced elevation in the gene expression of acetylcholine esterase, Glut3, insulin and cholinergic receptors in the cerebellum of diabetic rats. SIGNIFICANCE: Our studies suggest that curcumin plays a vital role in regulating the activity of cholinergic and insulin receptors and mechanism of glucose transportation through Glut3, which results in normalizing the diabetes-mediated cerebellar disorders. Thus, curcumin has a significant role in a therapeutic application for the prevention or progression of diabetic complications in the cerebellum. PMID- 19804786 TI - Lysophospholipids enhance taurine release from rat retinal vascular endothelial cells under hypoosmotic stress. AB - Sphingosine 1-phosphate (S1P) and lysophosphatidic acid (LPA) are simple bioactive lysophospholipids which exhibit an effect on blood vessels via their G protein-coupled receptors. The purpose of this study was (i) to clarify the impact of S1P and LPA on the release of taurine as an organic osmolyte from rat retinal vascular endothelial cells (RVECs) under hypoosmotic stress and (ii) to quantify the gene expression levels of S1P and LPA receptors in RVECs. When cultured RVECs (TR-iBRB2 cells) that had been preloaded with [(3)H]taurine were exposed to hypotonic buffer (230 mOsm) for 1 to 10 min, [(3)H]taurine release from the cells was several times greater than that using an isotonic buffer (300 mOsm). S1P and LPA significantly enhanced the [(3)H]taurine release under hypotonic conditions in a time- and concentration-dependent manner, whereas S1P and LPA had no significant effect under isotonic conditions. Quantitative real time PCR revealed that freshly isolated RVECs predominantly express mRNAs for S1P(1), S1P(4) and LPA(4). The S1P-enhanced [(3)H]taurine release under hypoosmotic conditions was significantly inhibited by an S1P(1) receptor antagonist. Inhibitor of the small GTPase Rho, C3 exotoxin, attenuated S1P- and LPA-enhanced [(3)H]taurine release. These results suggest that S1P and LPA play a novel role in the regulation of osmolyte efflux from RVECs in response to hypoosmotic stress via the activation of their specific receptors. PMID- 19804787 TI - Washout of small molecular contrast agent in carcinoma-derived experimental tumors. AB - The use of contrast-enhanced magnetic resonance imaging (MRI) for the assessment of breast carcinomas reveals satisfactory sensitivity, but due to low specificity, it does not obviate the need for subsequent tissue sampling. Its capability to differentiate benign from malignant lesion is under continuous investigation. Dynamic contrast-enhanced MRI (DCE-MRI) could improve specificity of MRI through the analysis of the kinetic of contrast enhancement. In particular, the study of the washout pattern is considered a promising tool to improve in vivo diagnosis and even to evaluate the response under chemotherapy. To provide a comprehensive characterization of this parameter in malignant tumor models, in vivo mapping of the washout of small molecular contrast agent (Gd DTPA, molecular weight 0.57 kDa) was carried out in three transplanted/spontaneous mammary tumors, which differed in their histopathological and microvascular features. It resulted that in all models around 40% of tumor volume lacks efficient washout; washout areas are frequently, but not always, restricted to the tumor periphery and that non-washout areas are not restricted to necrotic regions. Difference in the distribution of lymphatic vessels characterized spontaneous vs. transplanted tumors but did not produce a corresponding different washout pattern, confirming that Gd-DTPA drainage does not mainly depend on lymphatic architecture. Finally, the efficiency of washout is correlated with parameters obtainable during the earlier phases of the enhancement curve and in malignant tumors it could be indirectly estimated from them. PMID- 19804788 TI - Conflict processing of symbolic and non-symbolic numerosity. AB - It is commonly assumed that the processing of magnitudes occurs independent of modality or notation. Several studies have reported similar behavioural as well as neurophysiological responses to magnitudes presented in distinct modalities as well as notations, but a direct assessment of possible interactions between different modalities and notations, using measures of electro-cortical processing, is lacking. The present study investigates whether the neural activity underlying symbolic and non-symbolic numerosity processing interacts with the neural activity underlying physical size processing before, or proceeds independently until, selective activation of the motor system. We used a symbolic (Arabic numbers) and non-symbolic (arrays of dots) size congruency task and instructed subjects to judge either the numerical or the physical size of the stimuli, while event related potentials were recorded. Longer reaction times as well as a decrease in accuracy were obtained for incongruent compared to congruent trials. For the event related potential data, this congruency effect was also found with respect to the latency of the P3 component reflecting an interaction at the level of stimulus evaluation. Moreover, incongruence delayed the stimulus-locked but not the response-locked lateralized readiness potential. Together these results suggest that, irrespective of notation, the interaction between different magnitudes occurs before selective response activation. PMID- 19804789 TI - KIBRA and CLSTN2 polymorphisms exert interactive effects on human episodic memory. AB - Individual differences in episodic memory are highly heritable. Several studies have linked a polymorphism in the gene encoding the KIBRA protein to episodic memory performance. Results regarding CLSTN2, the gene encoding the synaptic protein calsyntenin 2, have been less consistent, possibly pointing to interactions with other genes. Given that both KIBRA and CLSTN2 are expressed in the medial temporal lobe and have been linked to synaptic plasticity, we investigated whether KIBRA and CLSTN2 interactively modulate episodic memory performance (n=383). We replicated the beneficial effect of the KIBRA T-allele on episodic memory, and discovered that this effect increases with the associative demands of the memory task. Importantly, the memory-enhancing effect of the KIBRA T-allele was boosted by the presence of the CLSTN2 C-allele, which positively affected memory performance in some previous studies. In contrast, the presence of CLSTN2 C-allele led to reduced performance in subjects homozygous for the KIBRA C-allele. Overall, these findings suggest that KIBRA and CLSTN2 interactively modulate episodic memory performance, and underscore the need for delineating the interactive effects of multiple genes on brain and behavior. PMID- 19804790 TI - Thinking of God moves attention. AB - The concepts of God and Devil are well known across many cultures and religions, and often involve spatial metaphors, but it is not well known if our mental representations of these concepts affect visual cognition. To examine if exposure to divine concepts produces shifts of attention, participants completed a target detection task in which they were first presented with God- and Devil-related words. We found faster RTs when targets appeared at compatible locations with the concepts of God (up/right locations) or Devil (down/left locations), and also found that these results do not vary by participants' religiosity. These results indicate that metaphors associated with the divine have strong spatial components that can produce shifts of attention, and add to the growing evidence for an extremely robust connection between internal spatial representations and where attention is allocated in the external environment. PMID- 19804791 TI - Orienting attention to objects in visual short-term memory. AB - We measured electroencephalographic activity during visual search of a target object among objects available to perception or among objects held in visual short-term memory (VSTM). For perceptual search, a single shape was shown first (pre-cue) followed by a search-array, and the task was to decide whether the pre cue was or was not in the search-array. For search of VSTM, a search-array was shown first followed by a single shape (post-cue), and the task was to decide whether the post-cue was or was not in the previously displayed search-array. We focused on early lateralized electrical brain activity over posterior and temporal areas time-locked to search-arrays in pre-cue trials and to post-cues in post-cue trials. In Experiment 1, search-arrays were composed of two lateralized shapes, displayed in the upper/lower two quadrants of the monitor. In Experiment 2, search-arrays were composed of four shapes, displayed at the corners of an imaginary square centered on fixation. In pre-cue trials, we observed an N2pc of about equal amplitude and latency for search-arrays composed of two or four shapes. In post-cue trials, we observed N2pc-like activity with search-arrays composed of two shapes, that was however substantially attenuated with search arrays composed of four shapes. For many aspects, attending to a perceptual object was functionally and neurally analogous to attending to an object held in VSTM, suggesting that spatial selective attention biases search of objects during both ongoing perception and retention. PMID- 19804792 TI - Reduced specificity of autobiographical memory and aging: do the executive and feature binding functions of working memory have a role? AB - Autobiographical memory (AM) is built up from various kinds of knowledge, from general to specific, via generative processes. Aging seems to particularly affect the episodic autobiographical information while preserving information that is more semantic. However, the mechanism of this deficit has not yet been thoroughly tested in relation to working memory. This study is designed to investigate, in a group of 100 subjects, the relationships between age, accessibility to different levels of AM specificity, and two main components of working memory: the central executive and the episodic buffer. We used a new task composed of four embedded verbal autobiographical fluencies (VAF) - from low to highest specificity levels exploring lifetime periods, general events, specific events, and details, plus tasks exploring free recall of episodic AM and updating, shifting, inhibition, and feature binding in working memory. The results demonstrate that age-related difficulties increase with level of specificity of autobiographical knowledge, i.e., from semantic to episodic aspects. Moreover, regression analyses mainly show that increase in age-related deficit with level of specificity of AM is largely mediated by performance on executive functions (updating and inhibition) and to a lesser extent feature binding in working memory. The results confirm in episodic AM the executive/working memory aging hypothesis, and for the first time highlight the role of episodic buffer in associating the various different details of specific events that elicit the conscious recollection. PMID- 19804793 TI - Estradiol reduces anxiety- and depression-like behavior of aged female mice. AB - Beneficial effects of the ovarian steroid, 17beta-estradiol (E(2)), for affective behavior have been reported in young individuals, but less is known about the effects of E(2) among older individuals, and the capacity of older individuals to respond to E(2) following its decline. In the present study, the effects of acute E(2) administration to aged mice for anxiety-like and depression-like behaviors were investigated. Intact female C57BL/6 mice (N=18) that were approximately 24 months old were administered vehicle (sesame oil, n=9) or E(2) (10 microg, n=9) subcutaneously 1h prior to behavioral testing. Mice were tested for anxiety-like behavior (open field, elevated plus maze, mirror chamber, light-dark transition task, Vogel conflict task) and depression-like behavior (forced swim task). To assess the role of general motor behavior and coordination in these aged mice, performance in an activity monitor and rotarod task, and total entries made in tasks (open field, elevated plus maze, light-dark transition task) were determined. Mice administered E(2), compared to vehicle, demonstrated anti anxiety behavior in the open field, mirror chamber, and light-dark transition task, and anti-depressive-like behavior in the forced swim task. E(2) also tended to have anti-anxiety effects in the elevated plus maze and Vogel task compared to vehicle administration, but these effects did not reach statistical significance. E(2) did not alter motor behavior and/or coordination in the activity monitor, open field, or rotarod tasks. Thus, an acute E(2) regimen produced specific anti anxiety and anti-depressant effects, independent of effects on motor behavior, when administered to aged female C57BL/6 mice. PMID- 19804794 TI - Impact of progressive resistance training on lipids and lipoproteins in adults: another look at a meta-analysis using prediction intervals. AB - OBJECTIVE: Given recently developed prediction intervals (PIs) in which a random mean effect for a new study is estimated from meta-analytic data, we used the results from our previously published meta-analysis to calculate PIs for changes in lipids and lipoproteins as a result of progressive resistance training (PRT) in adults. METHODS: Twenty-nine studies representing 1329 men and women (676 exercise, 653 control) were included. The primary outcomes included total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), ratio of total cholesterol to high-density lipoprotein cholesterol (TC/HDL-C), non-high-density lipoprotein cholesterol (non-HDL-C,) low-density lipoprotein cholesterol (LDL-C), and triglycerides (TG). Separate PIs (95%) were calculated for all lipids and lipoproteins. RESULTS: The expected outcomes of a new study on this topic were as follows: TC, -5.5 (-24.0, 13.0) mg/dl; HDL-C, 0.7 (-8.9, 10.4) mg/dl; TC/HDL-C, 0.5 (-1.8, 0.8); non-HDL-C, -8.7 (-35.7, 18.3) mg/dl; LDL-C, -6.1 (-28.9, 16.4) mg/dl; TG, -8.1 (-34.5, 18.3) mg/dl. CONCLUSIONS: Caution may be warranted in recommending that PRT improves TC, HDL-C, TC/HDL-C, non-HDL-C, LDL-C, and TG in adults. Future research should continue to examine the effects of PRT on lipids and lipoproteins in adults so as to determine optimal programs and populations in which PRT may have a positive effect. PMID- 19804795 TI - High-throughput ambulatory assessment of digital reactive hyperemia: concurrent validity with known cardiovascular risk factors and potential confounding. AB - OBJECTIVES: Reactive hyperemia index (RHI) measurement through digital peripheral arterial tonometry (PAT) is proposed for cardiovascular disease (CVD) risk screening. We evaluated the concurrent validity of RHI, measured in a high throughput ambulatory setting, with known CVD risk factors and biomarkers. METHODS: PAT was included in the 2007 EADS/Augsburg (Germany) cohort follow-up. CVD risk factors (age, sex, hypertension, hyperlipidemia, diabetes, smoking, physical activity, prevalent coronary heart disease, family history, cholesterol, triglycerides, blood pressure (BP), waist-to-hip ratio (WHR)) and biomarkers (d dimers, fibrinogen, log(c-reactive protein)) were assessed. The relationship between RHI and CVD risk factors and biomarkers was evaluated using multivariate linear regression, controlling for potential confounders (time of day, time since subject's last meal, baseline heart rate, examiner). RESULTS: Of 926 subjects approached, 710 underwent PAT and 603 (mean age 44.9+/-10 years, 88.7% men) with complete data were included for analysis. RHI was significantly related to being female (beta=0.128, p=0.02), low-density lipoprotein cholesterol (beta=-0.001, p=0.02), systolic BP (beta=0.007, p<0.001), WHR (beta=-1.04, p<0.01), time of day (beta=-0.011, p=0.04) and time since last meal (beta=0.013, p<0.01). CONCLUSIONS: Concurrent validity was partially demonstrated, while the need to control for potential confounding was reinforced. Participation was high and may be higher in less time-constrained settings. PMID- 19804797 TI - How does achievement motivation influence mental effort mobilization? Physiological evidence of deteriorative effects of negative affects on the level of engagement. AB - We tested whether the effect of achievement motivation on effort is modulated by two possible factors of the motivational intensity theory (Wright and Kirby, 2001): perceived difficulty and maximally justified effort. Approach-driven (N=16) and avoidance-driven (N=16) participants were first instructed to perform a reaction time task to the best of their abilities. Next, the participants were instructed to consistently beat their performance standard established in the first condition. Approach-driven participants showed a stronger decrease of midfrequency band of heart rate variability, which was used as an index of mental effort, than avoidance-driven participants in the second instruction condition. Moreover, avoidance-driven participants showed a higher corrugator supercilii reactivity, which was used as an index of negative affects, than approach-driven participants in the second instruction condition. No difference of perceived difficulty between groups was observed. Results suggested that avoidance-driven participants developed negative affects in the second instruction condition decreasing the maximally justified effort and their level of engagement. PMID- 19804798 TI - Impulsive aggressiveness of pregnant women affects the development of the fetal heart. AB - Mounting evidence indicates that the development of the fetus is heavily influenced by the intra-uterine milieu during pregnancy, and that such influence may have life-long consequences for the individual. The intra-uterine milieu is not only influenced by nutritional factors, but also by maternal endocrine and autonomic activity. Such activity is prone to be affected by an individual's personality, but only little is known about influences of maternal personality on the development of the fetus. We tested pregnant women for their propensity for impulsive, uncontrollable outbursts of temper (referred to here as moderate Intermittent Explosive Disorder, mIED). After the women gave birth, we measured electrocardiograms (ECGs) from their newborn infants to compare ECGs between newborns of women with and without mIED. The data show that infants of women with mIED have larger QRS complexes in the electrocardiogram, and lower heart rate variability, compared to infants of women without mIED. These results reveal effects of maternal mIED on the fetal heart development. These effects may predispose the individual to increased risk for later cardio-vascular disease. The findings open perspectives for better risk prevention models for the unborn child. PMID- 19804799 TI - Application of caffeine reveals input frequency-dependent determination of signal traveling routes between primary and secondary visual cortices in rats. AB - Upon entering the neocortex, neural signals are required to select which neocortical circuits to propagate through. The present study focused attention on use-dependent selection of signal-traveling routes. Rat brain slices including primary visual cortex (Oc1) and the medial part of the secondary visual cortex (Oc2M) were prepared. Electrical stimulation was delivered to white matter in Oc1 and spatiotemporal aspects of traveling signals were observed using optical recording methods under caffeine application. With an interstimulus interval (ISI) of 4-8s, signals traveled horizontally along deep layers from Oc1 to Oc2M, climbed within Oc2M, then returned along layer II/III from Oc2M to Oc1. Conversely, with an ISI of 40-64s, signals climbed within Oc1 and traveled horizontally along layer II/III from Oc1 to Oc2M in parallel with signals traveling along deep layers. Pharmacological experiments with antagonists for ionotropic glutamate receptors revealed that signal-traveling routes under higher frequency stimulation were N-methyl-d-aspartate (NMDA) receptor activity dependent, while those at the lower-frequency were non-NMDA receptor activity dependent. These results suggest that neural circuits between Oc1 and Oc2M possess an input frequency-dependent gating system, in which signal-traveling routes might be affected by the relative balance of receptor activities between NMDA and non-NMDA receptors. PMID- 19804796 TI - kappa-Opioid receptor signaling and brain reward function. AB - The dynorphin-like peptides have profound effects on the state of the brain reward system and human and animal behavior. The dynorphin-like peptides affect locomotor activity, food intake, sexual behavior, anxiety-like behavior, and drug intake. Stimulation of kappa-opioid receptors, the endogenous receptor for the dynorphin-like peptides, inhibits dopamine release in the striatum (nucleus accumbens and caudate putamen) and induces a negative mood state in humans and animals. The administration of drugs of abuse increases the release of dopamine in the striatum and mediates the concomitant release of dynorphin-like peptides in this brain region. The reviewed studies suggest that chronic drug intake leads to an upregulation of the brain dynorphin system in the striatum and in particular in the dorsal part of the striatum/caudate putamen. This might inhibit drug-induced dopamine release and provide protection against the neurotoxic effects of high dopamine levels. After the discontinuation of chronic drug intake these neuroadaptations remain unopposed which has been suggested to contribute to the negative emotional state associated with drug withdrawal and increased drug intake. kappa-Opioid receptor agonists have also been shown to inhibit calcium channels. Calcium channel inhibitors have antidepressant-like effects and inhibit the release of norepinephrine. This might explain that in some studies kappa opioid receptor agonists attenuate nicotine and opioid withdrawal symptomatology. A better understanding of the role of dynorphins in the regulation of brain reward function might contribute to the development of novel treatments for mood disorders and other disorders that stem from a dysregulation of the brain reward system. PMID- 19804800 TI - Characterization of a highly pathogenic avian influenza H5N1 clade 2.3.4 virus isolated from a tree sparrow. AB - The spread of H5N1 virus into a wide range of avian and mammalian species may facilitate the adaptation of the virus to human populations. In the present study, a survey of avian influenza virus in tree sparrows was performed and a HPAI H5N1 virus (A/tree sparrow/Jiangsu/1/08) was isolated. The H5N1 virus was found to be a genotype V variant belonging to clade 2.3.4, which had newly emerged in southern China in 2005. Genetic analysis showed that it had a close relation with A/Jiangsu/1/07 (H5N1), which was thought to be responsible for a probable limited person-to-person transmission in a family cluster in China in 2007. Pathogencity studies showed that the virus was highly virulent when experimentally inoculated into the chickens, tree sparrow, and mouse. As clade 2.3.4, genotype V variant of H5N1 virus has been accounted for the most of human fatal cases in China during 2005-2008, the existence of such H5N1 variants in the tree sparrows highlights the potential threat of this type of wild bird infection to veterinary and public health. PMID- 19804801 TI - Moussa virus: a new member of the Rhabdoviridae family isolated from Culex decens mosquitoes in Cote d'Ivoire. AB - Characterization of arboviruses at the interface of pristine habitats and anthropogenic landscapes is crucial to comprehensive emergent disease surveillance and forecasting efforts. In context of a surveillance campaign in and around a West African rainforest, particles morphologically consistent with rhabdoviruses were identified in cell cultures infected with homogenates of trapped mosquitoes. RNA recovered from these cultures was used to derive the first complete genome sequence of a rhabdovirus isolated from Culex decens mosquitoes in Cote d'Ivoire, tentatively named Moussa virus (MOUV). MOUV shows the classical genome organization of rhabdoviruses, with five open reading frames (ORF) in a linear order. However, sequences show only limited conservation (12 33% identity at amino acid level), and ORF2 and ORF3 have no significant similarity to sequences deposited in GenBank. Phylogenetic analysis indicates a potential new species with distant relationship to Tupaia and Tibrogargan virus. PMID- 19804802 TI - Controlled release of protein drugs from newly developed amphiphilic polymer based microparticles composed of nanoparticles. AB - A novel formulation of biodegradable microparticles was developed for the sustained release of peptide and protein drugs. The microparticles were formed by the aggregation of protein nanoparticles through water-in-oil (W/O) emulsion lyophilization and subsequent solid-in-oil-in-water (S/O/W) emulsion-solvent evaporation. Amphiphilic copolymers were used as an emulsifier in the W/O emulsion and matrix of the microparticles. Among the various copolymers investigated, poly(lactide-co-glycolide)-grafted dextran (Dex-g-PLGA) was chosen as the best candidate on the basis of the encapsulation efficiency and in vitro release profile, the near zero-order release without a significant initial burst, of human growth hormone (hGH). The release rate of hGH was controllable by changing the composition of Dex-g-PLGA. The in vivo release studies using normal mice revealed that the plasma concentration of hGH was maintained for 1week without a significant initial burst. The enhancement of biological activity of hGH by sustained release was confirmed by measuring the IGF-1 concentration and body weight of hypophysectomized mice. These results suggest the high potential of the newly developed microparticles for the sustained release of biopharmaceuticals. PMID- 19804803 TI - pH-Sensitive degradable polymersomes for triggered release of anticancer drugs: a comparative study with micelles. AB - pH-Sensitive degradable polymersomes and micelles were prepared based on diblock copolymer of poly(ethylene glycol) (PEG) and an acid-labile polycarbonate, poly(2,4,6-trimethoxybenzylidenepentaerythritol carbonate) (PTMBPEC). Polymersomes of PEG(1.9k)-PTMBPEC(6k) revealed average sizes of 100-200 nm. The acetals of polymersomes, similar to those of PEG(5k)-PTMBPEC(5.8k) micelles, though stable at pH 7.4 were prone to fast hydrolysis at mildly acidic pH of 4.0 and 5.0, with half lives of 0.5 and 3d, respectively. The acetal hydrolysis resulted in significant size increase of polymersomes, to over 1000 nm in 24h at pH 4.0. Drug encapsulation studies revealed that polymersomes were able to simultaneously load paclitaxel (PTX, hydrophobic) and doxorubicin hydrochloride (DOX.HCl, hydrophilic), whereas micelles loaded PTX only. Notably, polymersomes showed lower drug loading efficiencies for PTX than micelles (30.0-37.7% versus 61.4-65.2%). The in vitro release studies demonstrated that release of PTX and DOX.HCl from polymersomes was highly pH-dependent, i.e. significantly faster drug release at mildly acidic pH of 4.0 and 5.0 compared to physiological pH. Furthermore, much higher release rates were observed for PTX release from the polymersomes compared to that from the micelles under otherwise the same conditions. These pH-sensitive nano-sized degradable polymersomes hold great promise for combination therapy for cancers. PMID- 19804804 TI - Controlled release of chlorhexidine from amorphous microporous silica. AB - A new system for the controlled release of the antiseptic chlorhexidine is presented. Amorphous microporous silica (AMS) excipient material was synthesized via an acid catalyzed sol-gel method and shaped as powder or coating. Chlorhexidine diacetate was introduced into the pores of the AMS silica via the incipient wetness impregnation method. This silica reservoir maintained a slow release of chlorhexidine over more than 7days. Chlorhexidine release was controlled by configurational diffusion in the AMS pores having free diameters of less than 1nm. The release of chlorhexidine was fine tuned by adapting particle size and pore diameter. Controlled release of chlorhexidine from an AMS coating on silicon wafer was demonstrated. PMID- 19804805 TI - Encapsulation of NF-kappaB decoy oligonucleotides within echogenic liposomes and ultrasound-triggered release. AB - Echogenic liposomes (ELIP) have additional promise, beyond diagnostic agents, as vehicles for delivering oligonucleotides (ODN), especially if the release of the agent can be triggered and its uptake can be enhanced by ultrasound application at a specific site. The purpose of this study was to co-encapsulate air and NF kappaB decoy ODN within ELIP allowing ultrasound to release encapsulated ODN from ELIP, and to accurately quantify release of encapsulated ODN from ELIP upon ultrasound application. FITC-labeled sense ODN (2 mM) was incorporated within ELIP using freeze/thaw method. Encapsulation efficiency of FITC-ODN was spectrofluorometrically analyzed by quenching fluorescence of unencapsulated FITC ODN using a complementary strand tagged with Iowa Black FQ-ODN. Quenching of FITC ODN (0.05 microM) with Iowa Black FQ-ODN (0.1 microM) was found to be efficient (92.4+/-0.2%), allowing accurate determination of encapsulated ODN. Encapsulation efficiency of ODN was 14.2+/-2.5% in DPPC/DOPC/DPPG/CH liposomes and 29.6+/-1.5% in DPPC/DOPE/DPPG/CH liposomes. Application of ultrasound (1 MHz continuous wave, 0.26 MPa peak-to-peak pressure amplitude, 60s.) to the latter formulation triggered 41.6+/-4.3% release of ODN from ODN-containing ELIP. We have thus demonstrated that ODN can be encapsulated into ELIP and released efficiently upon ultrasound application. These findings suggest potential applications to gene therapy for atherosclerosis as well as a variety of other diseases. PMID- 19804806 TI - Squalene: A natural triterpene for use in disease management and therapy. AB - Squalene is a natural lipid belonging to the terpenoid family and a precursor of cholesterol biosynthesis. It is synthesized in humans and also in a wide array of organisms and substances, from sharks to olives and even bran, among others. Because of its significant dietary benefits, biocompatibility, inertness, and other advantageous properties, squalene is extensively used as an excipient in pharmaceutical formulations for disease management and therapy. In addition, squalene acts as a protective agent and has been shown to decrease chemotherapy induced side-effects. Moreover, squalene alone exhibits chemopreventive activity. Although it is a weak inhibitor of tumor cell proliferation, it contributes either directly or indirectly to the treatment of cancer due to its potentiation effect. In addition, squalene enhances the immune response to various associated antigens, and it is therefore being investigated for vaccine delivery applications. Since this triterpene is well absorbed orally, it has been used to improve the oral delivery of therapeutic molecules. All of these qualities have rendered squalene a potentially interesting excipient for pharmaceutical applications, especially for the delivery of vaccines, drugs, genes, and other biological substances. This paper is the first review of its kind and offers greater insight into squalene's direct or indirect contribution to disease management and therapy. PMID- 19804807 TI - Mannan-binding lectin deficiency increases the risk of recurrent infections in children with Down's syndrome. AB - Down syndrome (DS) is the most frequent cause of intellectual disability worldwide. DS individuals present abnormalities in the immune system that include high susceptibility to recurrent infections (RI) as well as to autoimmune diseases. Respiratory tract infections remain one of the major causes of death in DS individuals. Mannan-binding lectin (MBL) functions as an opsonina and initiates the lectin complement pathway. MBL deficiency was shown to increase the susceptibility to different infectious diseases, notably by extracellular pathogens. In the present study, MBL circulating levels were evaluated in 150 children with DS from Brazil, to clarify whether MBL deficiency is associated with the presence of RI in these patients. According to the clinical history 30.7% (46/150) of the DS children experienced RI, and MBL deficiency was seen in 34.8% (16/46) of them compared with 13.5% (14/104) of the DS children without RI (p = 0.005, odds ratio = 3.43, 95% confidence interval = 1.5-7.85). Moreover, MBL deficiency was significantly associated with the occurrence of pneumonia when compared with DS without RI (37.5%, 12/32 vs. 13.5% 14/104, p = 0.005, odds ratio = 3.68, 95% confidence interval = 1.5-6.95). These findings demonstrated that MBL deficiency increases the susceptibility to RI in DS patients and that, in the future, they could potentially benefit from MBL therapy. PMID- 19804808 TI - Effects of venlafaxine and fluoxetine on lymphocyte subsets in patients with major depressive disorder: a flow cytometric analysis. AB - BACKGROUND: Studies have yielded conflicting results concerning flow cytometric lymphocyte analyses in patients with depression. Data about the effect of antidepressants on lymphocyte subsets are also contradictory. The aim of this study was to determine effects of venlafaxine versus fluoxetine on lymphocyte subsets in depressive patients. METHODS: Sixty-nine patients diagnosed with major depressive disorder (MDD) according to DSM-IV and 36 healthy controls are included in the study. Sixty-nine patients were randomized to take fluoxetine (FLX) (n=33) or venlafaxine (VEN) (n=36). Serum lymphocyte subsets included CD3, CD4, CD8, CD16/56, CD19, CD45, Anti-HLA-DR which were measured by flow cytometric analyses at baseline and 6 weeks after the start of treatment. The severity of depression was evaluated with Hamilton rating scale for depression. RESULTS: At baseline, patients with MDD had significantly lower CD16/56 ratio and higher CD45 ratio compared to the controls. Although numerically higher in the VEN treated patients, treatment response rates between the FLX (53%) and the VEN (75%) groups were not different statistically. CD45 values decreased significantly in the VEN group at the end of the 6 week treatment period whereas no difference was observed in the FLX group. By the 6th week, treatment responders showed a significantly higher CD16/56 ratio than non-responders. Baseline severity of depression and anxiety was positively correlated with baseline CD45 ratio and negatively correlated with baseline CD16/56 ratio. We did not observe consistent changes in the absolute number of circulating B or T cells, nor in the helper/inducer (CD4) or suppressor/cytotoxic (CD8) subsets. CONCLUSIONS: CD16/56 was lower in patients with MDD and increased in treatment responders at 6th week. CD45 ratio was higher in patients with MDD than healthy subjects; it decreased with antidepressant treatment and was positively correlated with the severity of depression. Antidepressant treatment contributes to immune regulation in patients with major depressive disorder. PMID- 19804809 TI - Toxicity and carcinogenicity studies of methylene blue trihydrate in F344N rats and B6C3F1 mice. AB - Methylene blue trihydrate has a variety of biomedical and biologically therapeutic applications. Groups of 50 male and 50 female rats and mice were administered methylene blue trihydrate in 0.5% aqueous methylcellulose solution by gavage at doses of 0, 5, 25, or 50mg/kg bw/day (rats) or 0, 2.5, 12.5, and 25mg/kg bw/day (mice), 5 days per week for 2 years. In rats survival of all dosed groups was similar to that of the vehicle controls, whereas mice exhibited a dose dependent increase in survival. Rats receiving 25 and 50mg/kg bw/day and mice receiving 25mg/kg bw/day developed mild anemia. The incidences of pancreatic islet cell adenoma and adenoma or carcinoma (combined) were increased in all dosed groups of male rats, but increases were statistically significant in 25mg/kg bw/day males only and the dose-response was non-linear. There was a corresponding increase in the incidence of pancreatic islet cell hyperplasia but statistically significant only in the 50mg/kg bw/day male rats. There were no significant increases in neoplastic transformation observed in the mice; however, positive trends were noted for adenoma or carcinoma (combined) of the small intestine and malignant lymphoma. PMID- 19804810 TI - Monitoring bacterial-demineralization of human dentine by electrochemical impedance spectroscopy. AB - OBJECTIVE: The purpose of this study was two-fold: (1) to monitor bacterial biofilm formation and bacteria-induced demineralization of dentine in situ by using electrochemical impedance spectrum (EIS); (2) to examine the relationship between EIS findings and changes in the chemical composition and ultrastructure of dentine during bacteria-induced demineralization. METHODS: In this study, dentine demineralization was induced by Streptococcusmutans (ATCC 25175) in the presence of sucrose in culture medium and was monitored using two EIS measurement systems (Type A with a working electrode and Type B without a working electrode). Scanning electron microscopy (SEM), field emission scanning electron microscopy (FESEM), energy dispersive X-ray (EDX) and X-ray diffraction (XRD) were employed to examine the morphology, element contents and crystallinity of hydroxyapatite (HAP) on the dentine surface. Transverse microradiography (TMR) was used to characterize the lesion depth and degree of mineral loss during demineralization. RESULTS: The resistance of the bulk dentine (R(d)) and the apparent resistance of dentine (R(a)) measured from the Type A and Type B EIS systems, respectively, decreased gradually with demineralization. The resistance of the biofilm formed on dentine surface was determined by fitting the EIS data with equivalent circuits. The presence of biofilm slightly increased R(a) of dentine before demineralization. However, the electrochemical behavior of biofilm did not affect the decreasing impedance of dentine with demineralization. The SEM, EDX, XRD and TMR results demonstrated that the surface and bulk dentine gradually became more porous due to the loss of minerals during demineralization, which in turn resulted in the decrease in R(d) and R(a) values obtained from EIS systems. CONCLUSIONS: This investigation highlighted EIS as a potential technique to monitor biofilm formation and bacterial-induced demineralization in situ. PMID- 19804811 TI - Core-shell quantum dots tailor the fluorescence of dental resin composites. AB - OBJECTIVES: We characterized the optical properties, such as absorbance and fluorescence, of dental resins containing quantum dots (QD). We also determined the doping level needed to obtain a broad and nearly flat emission spectrum that provides the perception of white color. METHODS: The samples studied were resin composites from Charisma (Heraeus Kulzer) prepared with CdSe/ZnS core-shell QD (0.05-0.77 mass%). RESULTS: The results showed that the fluorescence of dental resin composites can be tailored by using CdSe/ZnS core-shell quantum dots. CONCLUSION: QD core incorporation into dental resins allows the fabrication of restorative materials with fluorescence properties that closely match those of natural human teeth. PMID- 19804812 TI - Involvement of cAMP/Epac/PI3K-dependent pathway in the antiproteolytic effect of epinephrine on rat skeletal muscle. AB - Very little is known about the signaling pathways by which catecholamines exert anabolic effects on muscle protein metabolism, stimulating protein synthesis and suppressing proteolysis. The present work tested the hypothesis that epinephrine induced inhibition of muscle proteolysis is mediated through the cAMP/Epac/PI3K dependent pathway with the involvement of AKT and Foxo. The incubation of extensor digitorum longus (EDL) muscles from rats with epinephrine and/or insulin increased the phosphorylation of AKT and its downstream target Foxo3a, a well known effect that prevents Foxo translocation to the nucleus and the activation of proteolysis. Similar effects on AKT/Foxo signaling were observed in muscles incubated with DBcAMP (cAMP analog). The stimulatory effect of epinephrine on AKT phosphorylation was completely blocked by wortmannin (selective PI3K inhibitor), suggesting that the epinephrine-induced activation of AKT is mediated through PI3K. As for epinephrine and DBcAMP, the incubation of muscles with 8CPT-2Me-cAMP (selective Epac agonist) reduced rates of proteolysis and increased phosphorylation levels of AKT and Foxo3a. The specific PKA agonist (N6BZ-cAMP) inhibited proteolysis and abolished the epinephrine-induced AKT and Foxo3a phosphorylation. On the other hand, inhibition of PKA by H89 further increased the phosphorylation levels of AKT and Foxo3a induced by epinephrine, DBcAMP or 8CPT-2Me-cAMP. These findings suggest that the antiproteolytic effect of the epinephrine on isolated skeletal muscle may occur through a cAMP/Epac/PI3K dependent pathway, which leads to the phosphorylation of AKT and Foxo3a. The parallel activation of PKA-dependent pathway also inhibits proteolysis and seems to limit the stimulatory effect of cAMP on AKT/Foxo3a signaling. PMID- 19804813 TI - Regulation of mitochondrial biogenesis during myogenesis. AB - Pathways involved in mitochondrial biogenesis associated with myogenic differentiation are poorly defined. Therefore, C(2)C(12) myoblasts were differentiated into multi-nucleated myotubes and parameters/regulators of mitochondrial biogenesis were investigated. Mitochondrial respiration, citrate synthase- and beta-hydroxyacyl-CoA dehydrogenase activity as well as protein content of complexes I, II, III and V of the mitochondrial respiratory chain increased 4-8-fold during differentiation. Additionally, an increase in the ratio of myosin heavy chain (MyHC) slow vs MyHC fast protein content was observed. PPAR transcriptional activity and transcript levels of PPAR-alpha, the PPAR co activator PGC-1alpha, mitochondrial transcription factor A and nuclear respiratory factor 1 increased during differentiation while expression levels of PPAR-gamma decreased. In conclusion, expression and activity levels of genes known for their regulatory role in skeletal muscle oxidative capabilities parallel the increase in oxidative parameters during the myogenic program. In particular, PGC-1alpha and PPAR-alpha may be involved in the regulation of mitochondrial biogenesis during myogenesis. PMID- 19804814 TI - Obesity and corticosteroids: 11beta-hydroxysteroid type 1 as a cause and therapeutic target in metabolic disease. AB - The metabolic abnormalities found associated with high blood glucocorticoid levels (e.g. rare Cushing's syndrome) include insulin-resistance, visceral obesity, hypertension, dyslipidaemia and an increased risk of cardiovascular diseases. The same constellation of abnormalities is found in the highly prevalent idiopathic obesity/insulin-resistance (metabolic)-syndrome. It is now apparent that tissue-specific changes in cortisol metabolism explain these parallels rather than altered blood cortisol levels. Primary among these changes is increased intracellular glucocorticoid reactivation, catalysed by the enzyme 11beta-hydroxysteroid dehydrogenase type (HSD)-1 in obese adipose tissue. Liver, skeletal muscle, endocrine pancreas, blood vessels and leukocytes express 11beta HSD1 and their potential role in metabolic disease is discussed. The weight of evidence, much of it gained from animal models, suggests that therapeutic inhibition of 11beta-HSD1 will be beneficial in most cellular contexts, with clinical trials supportive of this concept. PMID- 19804815 TI - Sub-chronic administration of AM251, CB1 receptor antagonist, within the nucleus accumbens induced sensitization to morphine in the rat. AB - It seems that there is a cross-talk between the cannabinoid CB1 and opioid receptors in the process of sensitization to opiates. In present study, we tried to examine the effect of solely administration of AM251, a CB1 receptor antagonist, on conditioned place preference (CPP) by ineffective dose of morphine in the rat. 102 adult male albino Wistar rats were used in these experiments. Subcutaneous administration of morphine (0.5, 1, 2.5, 5, 7.5 and 10 mg/kg) induced CPP only at the doses of > or = 5 mg/kg. The dose of 0.5mg/kg of morphine was selected as the appropriate (ineffective) dose for induction of CPP in animals which were previously received AM251 (5, 25 and 125 ng/0.5 microl per side) once daily for three days as a sub-chronic administration or those that received a single dose on the test day. Bilateral intra-accumbal sub-chronic but not single administration of AM251 dose-dependently produced sensitization to morphine and induced CPP by ineffective dose of morphine (0.5 mg/kg) in the rat. Bilateral intra-accumbal administration of neither saline nor DMSO (0.5 microl/side) had effects on sensitization to morphine. Our findings indicated that CB1 receptors within the nucleus accumbens are involved in the sensitization to morphine in rats. PMID- 19804816 TI - Peripheral contributions to the mechanical hyperalgesia following a lumbar 5 spinal nerve lesion in rats. AB - Using lumbar 5 (L5) dorsal root rhizotomy-bearing rats, we examined the extent to which L5 spinal nerve lesion (SNL)-induced mechanical hyperalgesia was governed by two peripheral components, that is Wallerian degeneration (WD) and peripherally-propagating injury discharge (PID). The contribution of WD to SNL induced hyperalgesia was studied by excluding PID with lidocaine treatment that blocked nerve conduction temporarily, but completely at the time of injury, whereas PID was examined separately by using brief tetanic electrical stimulation of the spinal nerve mimicking PID. Following the disappearance of L5 rhizotomy induced transient hyperalgesia, L5 SNL resulted in long-lasting mechanical hyperalgesia as early as one day post-SNL despite a PID block, highlighting the role of WD. In a comparative experiment, a delayed onset of hyperalgesia (7 days) was measured in L3 rhizotomy-bearing rats following L3 SNL with a PID block, in which injured fiber (L3) was separated from intact fibers (L4 and L5) anatomically until they meet at the peripheral terminals, supporting the importance of interactions between degenerating and adjacent intact fibers for WD induced hyperalgesia. Tetanic electrical stimulation of decentralized L5 spinal nerve resulted in mechanical hyperalgesia developing within 1 day and persisting for 7 days. This hyperalgesia was prevented by lidocaine blockade of the L5 nerve, and was unaffected by lidocaine blockades of the central inputs from L3 and L4 fibers during L5 nerve stimulation, suggesting the mediation of PID induced hyperalgesia by sensitization, not activation, of peripheral terminals of adjacent intact afferents. The similar hyperalgesia was also observed following electrical stimulation of decentralized L3 spinal nerve. Prior elimination of L4 C-fibers by local capsaicin prevented hyperalgesia induced either by L5 SNL with a PID block or by L5 nerve stimulation. These results suggest that neighboring C afferents remaining intact after partial nerve injury play a critical role in the development of mechanical hyperalgesia through interaction with degenerating afferents, and also via peripheral sensitization by PID. PMID- 19804817 TI - Na(+)/Cl(-)/creatine transporter activity and expression in rat brain synaptosomes. AB - Creatine is involved in brain ATP homeostasis and it may also act as neurotransmitter. Creatine transport was measured in synaptosomes obtained from the diencephalon and telencephalon of suckling and 2 month-old rats. Synaptosomes accumulate [(14)C]-creatine and this accumulation was Na(+)- and Cl(-)-dependent and inhibited by high external K(+). The latter suggests that the uptake process is electrogenic. The kinetic study revealed a K(m) for creatine of 8.7 microM. A 100-fold excess of either non-labelled creatine or guanidinopropionic acid abolished NaCl/creatine uptake, whereas GABA uptake was minimally modified, indicating a high substrate specificity of the creatine transporter. The levels of NaCl/creatine transporter (CRT) activity and those of the 4.2 kb CRT transcript (Northern's) were higher in the diencephalon than in the telencephalon, whereas the 2.7 kb transcript levels were similar in both brain regions and lower than those of the 4.2 kb. These observations suggest that the 4.2 kb transcript may code for the functional CRT. CRT activity and mRNA levels were similar in suckling and adult rats. To our knowledge the current results constitute the first description of the presence of a functional CRT in the axon terminal membrane that may serve to recapture the creatine released during the synapsis. PMID- 19804818 TI - Blocking of conditioned taste avoidance induced by wheel running. AB - In Experiment 1, compared to non-reinforced presentation of a food stimulus (A- >no US), the association of a food stimulus with wheel running (A-->US) blocked subsequent avoidance of a distinctive flavor (X), when both the food and flavor were followed by wheel running (AX-->US). Experiment 2 replicated and extended the blocking effect, demonstrating that the amount of avoidance of X after AX- >wheel training depended on the correlation between A-alone trials and wheel running-the predictiveness of the A stimulus. The present study is the first to demonstrate associative blocking of conditioned taste avoidance (CTA) induced by wheel running and strongly implicates associative learning as the basis for this kind of avoidance. PMID- 19804819 TI - Control of hypoxia-induced tumor cell adhesion by cytophilic human catalase. AB - Hypoxia-induced reactive oxygen species (ROS)-mediated expression of a variety of genes in endothelial cells has been suggested to be involved in abnormal cell adhesion. To prevent this by accelerated binding of catalase to endothelial cells, human catalase (hCAT), an enzyme catalyzing the decomposition of hydrogen peroxide, was fused with three repeats of arginine-glycine-aspartic acid peptide or nona arginine peptide at the C-terminal to obtain hCAT-(RGD)3 and hCAT-R9, respectively. Human CAT and its derivatives were expressed in yeast Pichia pastoris and purified. The specific activity and secondary structure of hCAT (RGD)3 and hCAT-R9 were close to those of hCAT, but these derivatives showed higher binding to the mouse aortic vascular endothelial cell line MAEC than hCAT, indicating that they are cytophilic derivatives. Hypoxic treatment of MAEC increased the intracellular ROS level, the binding of mouse melanoma cells, and the activity of transcription factors, hypoxia inducible factor-1 and nuclear factor-kappaB. hCAT-(RGD)3 or hCAT-R9 efficiently inhibited these changes compared with hCAT. These results indicate that cytophilic hCAT-(RGD)3 and hCAT R9 are effective in inhibiting hypoxia-induced tumor cell adhesion to endothelial cells. PMID- 19804820 TI - Endothelin-1 induces p66Shc activation through EGF receptor transactivation: Role of beta(1)Pix/Galpha(i3) interaction. AB - Endothelin-1 (ET-1) is a vasoconstrictor peptide known to be a potent mitogen for glomerular mesangial cells. We have shown that ET-1 stimulates the adaptor protein p66Shc through Rac/Cdc42 guanine nucleotide exchange factor beta(1)Pix. In this study, we demonstrate that ET-1-induced serine phosphorylation of p66Shc is mediated through Galpha(i3). Pertussis toxin treatment of cells induced a significant decrease in the interaction between beta(1)Pix and ET(A)-R, and an increase in the binding of Galpha(i3) and G(beta1) to beta(1)Pix. Activation of heterotrimeric G proteins by AlF(4)(-) resulted in an increase of Galpha(i3) binding to beta(1)Pix, which was significantly disrupted in cells expressing beta(1)Pix dimerization deficient mutant, beta(1)PixDelta (602-611). In cells expressing beta(1)PixDelta (602-611), ET-1-induced p66Shc activation was also significantly decreased. Specific inhibition of EGF receptor by AG1478 blocked ET 1-induced p66Shc activation and the binding of p66Shc and Galpha(i3) to beta(1)Pix. Inhibition of Erk1/2 blocked p66Shc activation induced by ET-1. Altogether, our results indicate that ET-1 activates p66Shc through EGF receptor transactivation, leading to the activation of Galpha(i3), beta(1)Pix and Erk1/2. PMID- 19804821 TI - Effects of the flavonol quercetin on the bioavailability of simvastatin in pigs. AB - The influence of the dietary flavonol quercetin on the pharmacokinetics of the HMG-CoA reductase inhibitor simvastatin was investigated in pigs. Simvastatin (0.25mg/kg body weight) was orally administered to six pigs either without or with quercetin (10mg/kg). In addition, simvastatin was administered to three pigs that had received a diet supplemented with the flavonol over a period of 1 week. Daily quercetin intake was 10mg/kg in these animals. Co-ingestion of quercetin with the statin did not alter area under the concentration time curve (AUC(0- >infinity)), time to achieve maximum plasma concentration (t(max)) or half-life (t(1/2)) of simvastatin. However, there was a trend towards a reduction of the maximum plasma concentration (C(max)) of simvastatin when quercetin was administered concomitantly (P=0.06). As compared to controls, AUC(0-->infinity) of simvastatin was significantly decreased after feeding the quercetin supplemented diet for 1 week. The plasma ratio of simvastatin and its acid metabolite was neither altered by the concomitant quercetin ingestion nor by feeding of the flavonol over a period of 1 week. We conclude that chronic ingestion of high doses of the flavonol quercetin will decrease the bioavailability of simvastatin to a significant extent. PMID- 19804822 TI - The influence of drug-drug interaction and patients' characteristics on valproic acid's clearance in adults with epilepsy using nonlinear mixed effects modeling. AB - Monitoring valproic acid (VPA) concentrations is especially challenging due to its highly variable pharmacokinetics (PK) and complex interactions with other antiepileptic drugs. We used sparse routine therapeutic drug monitoring data (n=200) from 129 adults with epilepsy to develop a population PK model of VPA, and determine the factors that influence its clearance (CL/F). Patients were on mono VPA therapy, or were concomitantly treated with carbamazepine, phenobarbital, topiramate (TPR), lamotrigine or benzodiazepines. A one compartment model with first-order absorption and elimination was used to fit the concentration-time VPA data. Estimates generated by NONMEM indicated that VPA CL/F was influenced by the patients' body weight (increases with the 0.556 exponent), VPA daily dose (if it is greater than 1000 mg/day, CL/F increases by 43%), and co-therapy with TPR (lowering CL/F for 23%). The interindividual variability in VPA CL/F was modeled with exponentional error model. The estimated coefficient of variation was 31.9%, while the residual variability was 23.8% for the proportional and 13.2mg/l for the additive component. The model was validated in a separate set of 24 patients, and the predictive performance was evaluated, that indicated unbias and acceptable precision. This study confirms the interaction of VPA with TPR, which is presumably dependent on VPA dose. PMID- 19804823 TI - Pharmacokinetic evaluation and modeling of formulated levodopa intranasal delivery systems. AB - Levodopa (L-dopa), the metabolic precursor of dopamine, has primarily been used for the treatment of Parkinson's disease (PD) in combination with carbidopa (C dopa). This study aims to investigate the feasibility of L-dopa nasal delivery systems prepared using maleic acid solution containing 2-hydroxypropyl-beta cyclodextrin, and to develop pharmacokinetic models. Following oral or intravenous administration of L-dopa plus C-dopa and intranasal dosing of L-dopa in the presence and absence of C-dopa to the rat, the concentrations of L-dopa in plasma and brain were determined using HPLC. The pharmacokinetic profiles were analyzed using non-compartmental and compartmental modeling approaches. Simultaneous nonlinear regression was performed to improve the identifiability of model parameters. L-Dopa was rapidly absorbed into blood and brain. The absolute bioavailabilities of oral and nasal preparations containing C-dopa were 17.7 and 45.4%, respectively. C-dopa caused a 1.2-fold decrease in the elimination rate of L-dopa, indicating decreased metabolism. Although the half-life after nasal administration was relatively short (less than 30 min) in both blood and brain regardless of C-dopa addition, the systemic exposure was promising due to rapid absorption. In conclusion, the L-dopa nasal delivery system could be used as a good rescue therapy for PD patients who experience symptom fluctuation with oral L-dopa administration. PMID- 19804824 TI - Three isostructural solvates of finasteride and their solid-state characterization. AB - Three crystalline hemi-hydrate, channel solvates (classified as solvates from here on) of finasteride (N-(1,1-di-methylethyl)-3-oxo-4-aza-5 alpha-androst-1-ene 17beta-carboxamide) have been obtained and fully characterized. The acetone, methyl ethyl ketone (MEK), and toluene solvates of finasteride, described herein, were found to be isostructural and belong as additional members to a family of previously reported finasteride solvates. Vacuum drying at 85 degrees C for 1 day produced the metastable, anhydrous Form II of finasteride from all three solvated materials. PMID- 19804825 TI - Thromboxane A2 receptor-mediated epidermal growth factor receptor transactivation: involvement of PKC-delta and PKC-epsilon in the shedding of epidermal growth factor receptor ligands. AB - We examined thromboxane A(2) receptor (TP)-mediated transactivation of epidermal growth factor receptor (EGFR) through the shedding of EGFR ligands. A TP agonist U46619 caused the phosphorylation of EGFR in 1321N1 human astrocytoma cells, which was inhibited by an EGFR selective inhibitor AG1478 and by a disintegrin and metalloproteinase (ADAM) inhibitor TAPI-2, indicating TP stimulation caused the EGFR transactivation through the EGFR ligand shedding. Since 1321N1 cells expressed heparin-binding EGF (HB-EGF) mRNA, the mechanism of TP-mediated EGFR transactivation was examined in HEK293 cells expressing alkaline phosphatase conjugated HB-EGF and TP. U46619 caused the shedding of HB-EGF in a time- and concentration-dependent manner. The TP-mediated shedding was inhibited by a furin inhibitor CMK, TAP-2, dominant-negative G alpha(q), a G(q/11) inhibitor YM254890, and also by a non-selective PKC inhibitor GF109203X and PKC down-regulation, but not by a conventional PKC inhibitor Go6976. Furthermore, siRNAs of PKC-delta and PKC-epsilon inhibited U46619-induced HB-EGF shedding. Although BAPTA/AM had no effect on U46619-induced shedding of HB-EGF, EGTA inhibited it. These results suggest that TP-mediated EGFR transactivation is partially caused by shedding of HB-EGF, which involves furin and ADAM via novel types of PKCs (PKC-delta and PKC epsilon) through G alpha(q/11) proteins in an extracellular Ca(2+)-dependent manner. PMID- 19804826 TI - Protein kinase D stabilizes aldosterone-induced ERK1/2 MAP kinase activation in M1 renal cortical collecting duct cells to promote cell proliferation. AB - Aldosterone elicits transcriptional responses in target tissues and also rapidly stimulates the activation of protein kinase signalling cascades independently of de novo protein synthesis. Here we investigated aldosterone-induced cell proliferation and extra-cellular regulated kinase 1 and 2 (ERK1/2) mitogen activated protein (MAP) kinase signalling in the M1 cortical collecting duct cell line (M1-CCD). Aldosterone promoted the proliferative growth of M1-CCD cells, an effect that was protein kinase D1 (PKD1), PKCdelta and ERK1/2-dependent. Aldosterone induced the rapid activation of ERK1/2 with peaks of activation at 2 and 10 to 30 min after hormone treatment followed by sustained activation lasting beyond 120 min. M1-CCD cells suppressed in PKD1 expression exhibited only the early, transient peaks in ERK1/2 activation without the sustained phase. Aldosterone stimulated the physical association of PKD1 with ERK1/2 within 2 min of treatment. The mineralocorticoid receptor (MR) antagonist RU28318 inhibited the early and late phases of aldosterone-induced ERK1/2 activation, and also aldosterone-induced proliferative cell growth. Aldosterone induced the sub cellular redistribution of ERK1/2 to the nuclei at 2 min and to cytoplasmic sites, proximal to the nuclei after 30 min. This sub-cellular distribution of ERK1/2 was inhibited in cells suppressed in the expression of PKD1. PMID- 19804827 TI - Changes in sympathetic activity in prion neuroinvasion. AB - Prion diseases are neurodegenerative diseases affecting humans and animals in which the infectious agent or prion is PrP(res), a protease-resistant conformer of the cell protein PrP. The natural transmission route of prion diseases is peripheral infection, with the lymphoreticular system (LRS) and peripheral nerves being involved in animal models of scrapie neuroinvasion and human prion diseases. To study the effects of PrP neuroinvasion on sympathetic nerve function, we measured plasma catecholamine levels, blood pressure, heart rate, and PrP tissue levels in intraperitoneally or intracerebrally infected mice. The results indicate a specific alteration in sympathetic nerve function because the levels of noradrenaline (but not adrenaline) were increased in the animals infected peripherally (but not in those infected intracerebrally) and correlated with increased blood pressure. These findings confirm that prion neuroinvasion uses the sympathetic nervous system to spread from the periphery to the central nervous system after invading the LRS. PMID- 19804828 TI - Increased expression of glutamate transporters in subcortical white matter after transient focal cerebral ischemia. AB - Transient focal cerebral ischemia leads to extensive excitotoxic glial damage in the subcortical white matter. Efficient reuptake of released glutamate is essential for preventing glutamate receptor overstimulation and neuronal and glial death. The present study evaluates the expression of the main glutamate transporters (EAAT1, EAAT2, and EAAT3) in subcortical white matter of the rat after transient middle cerebral artery occlusion. Western blot analysis and immunohistochemistry show an increase in the expression of EAAT1 and EAAT2 in subcortical white matter early after ischemia which subsequently decreases at longer reperfusion periods. However, expression of both EAAT1 and EAAT2 remains higher in astrocytes forming the gliotic scar and in microglial/macrophage cells at the border of or within the infarct area, respectively. Taken together, these results indicate that there is a transient enhanced expression of EAATs in the subcortical white matter early after ischemia. Our findings reveal an adaptive response of subcortical white matter to increased levels of glutamate during focal cerebral ischemia which may limit excitotoxic damage. PMID- 19804829 TI - 1-(2',4'-dichlorophenyl)-6-methyl-N-cyclohexylamine-1,4-dihydroindeno[1,2 c]pyrazole-3-carboxamide, a novel CB2 agonist, alleviates neuropathic pain through functional microglial changes in mice. AB - Neuropathic pain is a devastating neurological disease that seriously affects quality of life in patients. The mechanisms leading to the development and maintenance of neuropathic pain are still poorly understood. However, recent evidence points towards a role of spinal microglia in the modulation of neuronal mechanisms. In this context, cannabinoids are thought to modulate synaptic plasticity as well as glial functions. Here, we have investigated the effect of chronic treatment with a selective agonist of cannabinoid type 2 receptor (CB2), 1-(2',4'-dichlorophenyl)-6-methyl-N-cyclohexylamine-1,4-dihydroindeno[1,2 c]pyrazole-3 carboxamide (NESS400), on pain thresholds in the spared nerve injury (SNI) model in the mouse and on the distribution and activation of spinal microglia. Repeated treatment with NESS400 (4 mg/kg) significantly alleviated neuropathic mechanical allodynia and thermal hyperalgesia. In the dorsal horn (L4 L6) of neuropathic mice microglia activation (quantification of the length of microglial processes) and astrocytosis were associated with CB2 receptor over expression on both cell types. Treatment with NESS400 significantly reduced the number of hypertrophic microglia while leaving microglial cell number unaffected and reduced astrogliosis. Moreover, prolonged administration of NESS400 reduced mRNA expression of pro-inflammatory markers and enhanced anti-inflammatory marker gene expression in dorsal horn extracts. In conclusion, we show that selective CB2 receptor stimulation prevents thermal hyperalgesia, alleviates mechanical allodynia and facilitates the proliferation of anti-inflammatory microglial phenotype in the ipsilateral dorsal horn of the spinal cord in SNI mice. PMID- 19804830 TI - Remodeling of striatal NMDA receptors by chronic A(2A) receptor blockade in Huntington's disease mice. AB - Excitotoxicity plays a major role in the pathogenesis of Huntington disease (HD), a fatal neurodegenerative disorder. Adenosine A(2A) receptors (A(2A)Rs) modulate excitotoxicity and have been suggested to play a pathogenetic role in HD. The main aim of this study was to evaluate the effect of A(2A)R blockade on the expression and functions of NMDA receptors in the striatum of HD mice (R6/2). We found that 3 weeks' treatment with SCH 58261 (0.01 mg/kg/day i.p. from the 8th week of age) modified NR1 and NR2A/NR2B expression in the striatum of R6/2 (Western blotting) while had no effect on NMDA-induced toxicity in corticostriatal slices (electrophysiological experiments). In conclusion, in vivo A(2A)R blockade induced a remodeling of NMDA receptors in the striatum of HD mice. Even though the functional relevance of the above effect remains to be fully elucidated, these results add further evidence to the modulatory role of A(2A)Rs in HD. PMID- 19804832 TI - The blood-brain barrier. PMID- 19804831 TI - Network perspectives on the mechanisms of deep brain stimulation. AB - Deep brain stimulation (DBS) is an established medical therapy for the treatment of movement disorders and shows great promise for several other neurological disorders. However, after decades of clinical utility the underlying therapeutic mechanisms remain undefined. Early attempts to explain the mechanisms of DBS focused on hypotheses that mimicked an ablative lesion to the stimulated brain region. More recent scientific efforts have explored the wide-spread changes in neural activity generated throughout the stimulated brain network. In turn, new theories on the mechanisms of DBS have taken a systems-level approach to begin to decipher the network activity. This review provides an introduction to some of the network based theories on the function and pathophysiology of the cortico basal-ganglia-thalamo-cortical loops commonly targeted by DBS. We then analyze some recent results on the effects of DBS on these networks, with a focus on subthalamic DBS for the treatment of Parkinson's disease. Finally we attempt to summarize how DBS could be achieving its therapeutic effects by overriding pathological network activity. PMID- 19804833 TI - Targeting ERK1/2 activation and proliferation in human primary schwannoma cells with MEK1/2 inhibitor AZD6244. AB - Deficiency of the tumor suppressor merlin leads to the development of multiple tumors of the nervous system, such as schwannomas, meningiomas, and ependymomas. Due to the benign character of these tumors, classical chemotherapy is ineffective. Current therapies, surgery, and radiosurgery are local and quite invasive, thus new systemic treatments are required. We have previously described the Raf/mitogen-activated kinase (MEK)/extracellular signal-regulated kinase (ERK) pathway activation and its role in schwannoma growth. Here, we targeted MEK1/2 known as a convergence point for multiple cascades towards ERK1/2 activation and cell proliferation, using MEK1/2 inhibitor AZD6244 (ARRY-142886; Astra Zeneca). We show that AZD6244 at low concentration completely abolished platelet-derived growth factor-DD-mediated ERK1/2 activation and cell proliferation in human primary schwannoma cells. Moreover, this drug was not toxic for either schwannoma or Schwann cells and has been reported to be safe with tolerable side effects. Thus, AZD6244 can be considered as a drug candidate for schwannoma treatment. PMID- 19804834 TI - Contribution of reactive oxygen species to migration/invasion of human glioblastoma cells U87 via ERK-dependent COX-2/PGE(2) activation. AB - In the presence of 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulation, an increase in the migration/invasion of U87 glioblastoma cells was detected by a wound healing assay, transwell analysis, and spheroid formation assay by inducing matrix metalloproteinase-9 (MMP-9) enzyme activity via a gelatin zymographic analysis. A dose- and time-dependent increase in cyclooxygenase-2 (COX-2) gene expression with elevated prostaglandin E(2) (PGE(2)) production was identified in TPA- but not in 4alpha-TPA (a respective inactive compound)-treated U87 cells TPA induced migration/invasion was significantly blocked by adding the COX-2-specific inhibitor, NS398, through a reduction in PGE(2) production. Data from the pharmacological studies using specific chemical inhibitors showed that activation of protein kinase C (PKC) and extracellular signal-regulated kinases (ERKs) was involved in TPA-induced migration/invasion, COX-2 protein expression, and MMP-9 activation. Stimulation of intracellular peroxide production by TPA was detected by a DCHF-DA assay, and the addition of superoxide dismutase (SOD) or tempol significantly inhibited TPA-induced migration/invasion and COX-2 protein expression accompanied by a decrease in peroxide production. An increase in NADPH oxidase activity by TPA was examined, and TPA-induced migration/invasion was blocked by adding DPI, an NADPH oxidase inhibitor. Additionally, the natural flavonoids quercetin (QE), baicalein (BE), and myricetin (ME) effectively blocked TPA-induced migration/invasion while simultaneously inhibiting COX-2/PGE(2) production, MMP-9 enzyme activity, and peroxide production in U87 cells. The contribution of ROS production to the migration/invasion of U87 glioblastoma cells via ERK-activated COX-2/PGE(2) and MMP-9 induction was first investigated here, and agents such as QE, BE, and ME with the ability to block these events possess the potential to be developed for use against migration/invasion by glioblastomas. PMID- 19804835 TI - Sequence and expression analysis of extracellular signal-regulated kinase 1 from flounder (Paralichthys olivaceous). AB - Extracellular signal-regulated kinases (ERKs) are a subgroup of mitogen-activated protein kinases (MAPK) that function as important intermediates in signal transduction pathways initiated by several types of cell surface receptors. We cloned a transcript of ERK1 from a cDNA library of flounder leukocytes stimulated with bacterial lipopolysaccharide and hemagglutinin lectin. Flounder ERK1 consists of 1502 nucleotides and encodes a polypeptide of 393 amino acids. Flounder ERK1 showed 90 and 89% amino acid sequence identity to ERK1 of carp and zebrafish, respectively, and over 85% to that of mammals. Multiple bands were detected by Southern blot analysis of flounder genomic DNA after digestion with various restriction enzymes, implying the presence of additional MAPK genes in flounder. Real-time PCR revealed the ubiquitous expression of flounder MAPK in all tissues with high levels of transcription in brain, gill, and fin, but not in muscle or skin. Flounder MAPK was successfully expressed in mammalian COS1 cells and phosphorylated myelin basic protein (MBP) substrate when the cells were stimulated with PMA or EGF, indicating that flounder MAPK is functional in animal cells. PMID- 19804836 TI - Tease or threat? Judging social interactions from bodily expressions. AB - We casually observe many interactions that do not really concern us. Yet sometimes we need to be able to rapidly appraise whether an interaction between two people represents a real threat for one of them rather than an innocent tease. Using functional magnetic resonance imaging, we investigated whether small differences in the body language of two interacting people are picked up by the brain even if observers are performing an unrelated task. Fourteen participants were scanned while watching 3-s movies (192 trials and 96 scrambles) showing a male person either threatening or teasing a female one. In one task condition, observers categorized the interaction as threatening or teasing, and in the other, they monitored randomly appearing dots and categorized the color. Our results clearly show that right amygdala responds more to threatening than to teasing situations irrespective of the observers' task. When observers' attention is not explicitly directed to the situation, this heightened amygdala activation goes together with increased activity in body sensitive regions in fusiform gyrus, extrastriate body area-human motion complex and superior temporal sulcus and is associated with a better behavioral performance of the participants during threatening situations. In addition, regions involved in action observation (inferior frontal gyrus, temporoparietal junction, and inferior parietal lobe) and preparation (premotor, putamen) show increased activation for threat videos. Also regions involved in processing moral violations (temporoparietal junction, hypothalamus) reacted selectively to the threatening interactions. Taken together, our results show which brain regions react selectively to witnessing a threatening interaction even if the situation is not attended because the observers perform an unrelated task. PMID- 19804837 TI - T helper17 cells are sufficient but not necessary to induce acute graft-versus host disease. AB - T helper (Th)1 cells were considered responsible for the induction of graft versus-host disease (GVHD), but recently the concept has been challenged. Th17 cells play a critical role in mediating autoimmune diseases, but their role in the pathogenesis of GVHD remains unclear. Herein we compare the ability of in vitro generated Th1 and Th17 cells from C57BL/6 mice to induce GVHD in lethally irradiated BALB/c recipients. Allogeneic Th17 cells had superior expansion and infiltration capabilities in GVHD target organs, which correlated with their increased pathogenicity when compared with naive or Th1 controls. Th17 cells caused no pathology in the syngeneic recipients, indicating that antigen activation was required for their pathogenicity. Polarized Th17 cells could not maintain their phenotype in vivo as they produced a significant amount of interferon (IFN)-gamma after being transplanted into allogeneic recipients; however, IFN-gamma was not required for Th17 cell-induced GVHD. Further, we evaluated the pathogenesis of Th17 cells in GVHD by using polyclonal nonprimed CD4T cells in a clinically relevant allogeneic bone marrow transplantation (BMT) setting. We found that disruption of Th17-differentiation alone by targeting RORgammat (Th17-specific transcription factor) had no significant effect on GVHD development. We conclude that Th17 cells are sufficient but not necessary to induce GVHD. PMID- 19804838 TI - Docosahexaenoic acid neurolipidomics. AB - Mediator lipidomics is a field of study concerned with the characterization, structural elucidation and bioactivity of lipid derivatives actively generated by enzymatic activity. It is well known that omega-3 fatty acids are beneficial for brain function. Docosahexaenoic acid [DHA; 4 22:6(n-3)] is the most abundant essential omega-3 fatty acid present in the brain and it has multiple mechanisms of exerting protective effects after cellular injury. Certain lipid species produced from DHA early during the reperfusion stage of brain ischemia reperfusion injury are generated in order to help the cell cope as the injury progresses. We explore these newly discovered lipid mediators in order to understand their role in the cell. We have identified one of these potentially protective lipid mediators as a novel stereospecific DHA-derived fatty acid, called neuroprotectin D1 (NPD1; 10R,17S-dihydroxy-docosa-4Z,7Z,11E,15E,19Z hexaenoic acid). DHA also has important roles in pro-survival signaling cascades after ischemia-reperfusion in injury. It has been shown to accelerate AKT translocation and activation and has binding affinity with an important PPAR gamma family of ligand-activated nuclear receptors that have been implicated in various aspects of lipid metabolism and have been shown to have anti-inflammatory actions. Here we present an overview of these mechanisms and discuss the potential of using DHA signaling in the development of treatments for the large population of patients suffering from the devastating consequences of stroke. PMID- 19804839 TI - A cell-based assay for screening lipoxygenase inhibitors. AB - Lipoxygenases (LOX) form a family of lipid peroxidizing enzymes within the plant and animal kingdoms. In humans, six functional lipoxygenase isoforms have been identified. 5-LOX, "platelet-type" 12-LOX (p12-LOX) and 15-LOX type 1 (15-LOX1), originally identified in leukocytes, platelets, and reticulocytes, respectively, generate lipid mediators involved in host cellular functions and in the pathophysiology of asthma, cardiovascular diseases, and cancer. The pharmaceutical industry has reinvigorated their programs to develop novel LOX inhibitors in view of recent findings. However, high throughput LOX screening assays to test novel agents against these intracellular enzymes are limited. We describe a cell-based 96-well microplate fluorescence assay tested against several existing LOX inhibitors, and validate the assay by comparing known IC(50) values and HPLC analysis, which may provide a useful screen for novel LOX inhibitors. PMID- 19804840 TI - High incidence of colonic perforation during colonoscopy in hemodialysis patients with end-stage renal disease. AB - BACKGROUND & AIMS: Colonic perforation is a rare but life-threatening complication of colonoscopy. We evaluated the incidence of colonic perforation that resulted from colonoscopy in patients who underwent hemodialysis compared with those who did not have this procedure (controls). METHODS: Data from a total of 15,098 consecutive patients who underwent colonoscopy from January 2001 to December 2008 in Nagoya Kyoritsu Hospital were analyzed retrospectively. Patients were divided into 2 groups: 1106 hemodialysis patients and 13,992 controls. The incidence of colonic perforation, patient characteristics, and locations of perforation during colonoscopy were compared between the 2 groups. Furthermore, perforated mucosa samples from colons were examined by pathology analysis. RESULTS: Colonic perforations occurred in 5 hemodialysis patients and 3 controls. The incidence of colonic perforation was markedly higher in the hemodialysis group than in the control group (0.45% vs 0.02%; odds ratio, 21.17; 95% confidence interval, 5.05-88.73; P < .0001). Even after multivariate analysis of age, sex, and patients who received polypectomies, hemodialysis still was associated independently with the risk of colonic perforation during colonoscopy (odds ratio, 19.91; 95% confidence interval, 4.61-85.93; P < .0001). Pathologic examination of perforated mucosa was performed in 3 hemodialysis patients and 3 control patients. beta2-microglobulin deposition was observed in all 3 hemodialysis patients. In contrast, beta2-microglobulin deposition was not detected in control patients. CONCLUSIONS: There is a higher risk of colonic perforation during colonoscopy among patients who received hemodialysis compared with those who did not. beta2-microglobulin deposition might have a role in perforation in patients who receive hemodialysis. PMID- 19804841 TI - Hepatitis B virus load in serum does not reflect histologic activity in patients with decompensated cirrhosis. AB - BACKGROUND & AIMS: Little is known about whether histologic data can predict which patients with hepatitis B virus (HBV)-related decompensated cirrhosis will respond to antiviral therapies. We assessed the relationship between serum HBV DNA load and histologic activity by analyzing liver specimens from patients with decompensated cirrhosis. METHODS: The study included 72 consecutive patients who underwent liver transplantation for HBV-related decompensated cirrhosis between November 2000 and March 2008. None of the patients had received nucleoside or nucleotide analogues more than 2 weeks before transplantation. Serum HBV DNA levels at the time of transplantation were compared with histologic activity in explanted liver specimens. RESULTS: The median HBV DNA level of the 72 patients was 5.40 log(10) copies/mL (range, 1.45-8.00 log(10) copies/mL). There were no differences in HBV DNA level between patients grouped according to lobular or portoperiportal activity (P = .678, P = .291, respectively). Of 16 patients (22.2%) with HBV DNA levels less than 2000 copies/mL, 8 patients (50.0%) had moderate or severe portoperiportal activity; their median alanine aminotransferase level was 30.5 U/L (range, 12-135 U/L). CONCLUSIONS: HBV DNA load does not reflect histologic activity in patients with HBV-related decompensated cirrhosis. Although patients with decompensated cirrhosis might have normal levels of alanine aminotransferase and a low level of viremia (<2000 copies/mL), they still can have significant portoperiportal activity. PMID- 19804842 TI - Sequential and concomitant therapy with four drugs is equally effective for eradication of H pylori infection. AB - BACKGROUND & AIMS: Sequential therapy with a proton pump inhibitor (PPI) and amoxicillin followed by a PPI, clarithromycin, and an imidazole agent reportedly have a better rate of curing Helicobacter pylori infection than PPI, amoxicillin, and clarithromycin triple therapy. The concomitant administration of these 4 drugs (concomitant therapy) is also an effective treatment strategy. We compared the efficacies of sequential and concomitant therapy and analyzed the effects of antibiotic resistance in patients with H pylori infection. METHODS: In a randomized trial of 232 H pylori-infected patients from 3 hospitals in Kaohsiung, Taiwan, patients were given 10 days of sequential (n = 115) or concomitant (n = 117) therapy. H pylori status was confirmed by endoscopy or urea breath test. RESULTS: Intention-to-treat analysis demonstrated similar eradication rates for sequential (92.3%; 95% confidence interval [CI], 87.5%-97.1%) and concomitant therapy (93.0%; 95% CI, 88.3%-97.7%)(P = .83). Per-protocol eradication results were similar for sequential (93.1%; 95% CI, 90.7%-95.5%) and concomitant therapy (93.0%; 95% CI, 88.3%-97.7%) (P = .99). Univariate analysis showed that compliance and resistance to clarithromycin were independent determinants of eradication. Dual resistance did not influence the level of eradication in the concomitant group, but significantly affected that of the sequential therapy group. Clarithromycin resistance was less frequent than expected. CONCLUSIONS: Sequential or concomitant therapy with a PPI, amoxicillin, clarithromycin, and an imidazole agent are equally effective and safe for eradication of H pylori infection. Resistance to clarithromycin, compliance, and adverse events reduced the level of eradication. Concomitant therapy may be more suitable for patients with dual resistance to antibiotics. PMID- 19804843 TI - Image of the month. Pancreatic cystosis complicating cystic fibrosis. PMID- 19804844 TI - Surprise finding at endoscopic retrograde cholangiopancreatography. PMID- 19804845 TI - Group differences in proneness to inflammation. AB - All humans are primarily descendants from a diaspora out of Africa approximately 50,000 years ago although there are some indications of admixture with local populations of archaic humans outside Africa. The burden of infectious disease is greater in tropical Africa than elsewhere on earth in historic times, and it was less outside Africa, especially in the New World where passage through the Beringian filter kept many Old World parasites from entering the New World with humans. As a consequence we expect that the immune system, especially susceptibility to inflammation, will be "tuned up" in people with recent tropical African ancestry, intermediate in people of European and Asian ancestry, and perhaps "tuned down" in people of Native American ancestry. We suggest that evolved responses to different pathogen burdens among geographic groups may contribute to higher rates of inflammatory disease in modern people. PMID- 19804846 TI - Interleukin-6 subfamily cytokines and rheumatoid arthritis: role of antagonists. AB - Many cytokines have been implicated in the inflammatory pathways that characterize rheumatoid arthritis (RA) and related inflammatory diseases of the joints. These include members of the interleukin-6 (IL-6) family of cytokines, several of which have been detected in excess in the synovial fluid from RA patients. What makes the IL-6 group of cytokines a family is their common use of the glycoprotein 130 (gp130) receptor subunit, to which they bind with different affinities. Several strategies have been developed to block the pro-inflammatory activities of IL-6 subfamily cytokines. These include the application of monoclonal antibodies, the creation of mutant form(s) of the cytokine with enhanced binding affinity to gp130 receptor and the generation of antagonists by selective mutagenesis of the specific cytokine/gp130 receptor-binding site(s). The rationale for the use of anti-cytokine therapy in inflammatory joint diseases is based on evidence from studies in vitro and in vivo, which implicate major cytokines such as interleukin-1 (IL-1), tumour necrosis factor (TNF)-alpha and IL 6 in RA pathogenesis. In particular, IL-6 subfamily antagonists have a wide range of potential therapeutic and research applications. This review focuses on the role of some of the IL-6 subfamily cytokines in the pathogenesis of the inflammatory diseases of the joints (IJDs), such as RA. In addition, an overview of the recently developed antagonists will be discussed. PMID- 19804847 TI - Glucans from the Caripia montagnei mushroom present anti-inflammatory activity. AB - Caripia montagnei is a basidiomycete species which contains polysaccharides with immunomodulatory properties. An extract of this mushroom underwent removal of the fat content by organic solvent and subsequently proteolysis. The aqueous phase obtained after proteolysis was precipitated with methanol yielding a fraction containing carbohydrates (98.7+/-3.3%) and protein (1.3+/-0.25%). Chemical analysis, infrared spectroscopy and nuclear magnetic resonance (NMR) showed that the carbohydrate fraction contained (63.3+/-4.1) of beta-glucans and proteins (2.2+/-0.3%). These glucans (50mg/kg of body weight) significantly reduced the inflammatory infiltrate produced by thioglycolate-induced peritonitis by 75.5+/ 5.2%, when compared to Wy-14643 (60.3+/-6.1%), PFOA (37.8+/-2.8%) and clofibrate (52.2+/-3.2%), p<0.001, which are of the peroxisome proliferator-activated receptor (PPAR-alpha). L-NAME, a nitric oxide synthase inhibitor, reduced the plantar edema in Wistar rats by 91.4+/-1.3% (p<0.001). A significant reduction in nitric oxide (NO) levels was observed in the exudates when the glucans was used in comparison to carrageenan. The C. montagnei glucans did not present signs of inducing cytotoxicity. A decrease in IL-1ra, IL-10 and IFN-gamma in the peritonitis model was observed. Thus, the results suggest that glucans from the C. montagnei mushroom is an effective immunomodulator and may have potential for anti-inflammatory properties. PMID- 19804849 TI - Incidence of fragile X syndrome by newborn screening for methylated FMR1 DNA. AB - Fragile X syndrome (FXS) results from a CGG-repeat expansion that triggers hypermethylation and silencing of the FMR1 gene. FXS is referred to as the most common form of inherited intellectual disability, yet its true incidence has never been measured directly by large population screening. Here, we developed an inexpensive and high-throughput assay to quantitatively assess FMR1 methylation in DNA isolated from the dried blood spots of 36,124 deidentified newborn males. This assay displays 100% specificity and 100% sensitivity for detecting FMR1 methylation, successfully distinguishing normal males from males with full mutation FXS. Furthermore, the assay can detect excess FMR1 methylation in 82% of females with full mutations, although the methylation did not correlate with intellectual disability. With amelogenin PCR used for detecting the presence of a Y chromosome, this assay can also detect males with Klinefelter syndrome (KS) (47, XXY). We identified 64 males with FMR1 methylation and, after confirmatory testing, found seven to have full-mutation FXS and 57 to have KS. Because the precise incidence of KS is known, we used our observed KS incidence as a sentinel to assess ascertainment quality and showed that our KS incidence of 1 in 633 newborn males was not significantly different from the literature incidence of 1 in 576 (p = 0.79). The seven FXS males revealed an FXS incidence in males of 1 in 5161 (95% confidence interval of 1 in 10,653-1 in 2500), consistent with some earlier indirect estimates. Given the trials now underway for possible FXS treatments, this method could be used in newborn or infant screening as a way of ensuring early interventions for FXS. PMID- 19804848 TI - Familial hemophagocytic lymphohistiocytosis type 5 (FHL-5) is caused by mutations in Munc18-2 and impaired binding to syntaxin 11. AB - Rapid intracellular transport and secretion of cytotoxic granules through the immunological synapse requires a balanced interaction of several proteins. Disturbance of this highly regulated process underlies familial hemophagocytic lymphohistiocytosis (FHL), a genetically heterogeneous autosomal-recessive disorder characterized by a severe hyperinflammatory phenotype. Here, we have assigned FHL-5 to a 1 Mb region on chromosome 19p by using high-resolution SNP genotyping in eight unrelated FHL patients from consanguineous families. Subsequently, we found nine different mutations, either truncating or missense, in STXBP2 in twelve patients from Turkey, Saudi Arabia, and Central Europe. STXBP2 encodes syntaxin binding protein 2 (Munc18-2), involved in the regulation of vesicle transport to the plasma membrane. We have identified syntaxin 11, a SNARE protein mutated in FHL-4, as an interaction partner of STXBP2. This interaction is eliminated by the missense mutations found in our FHL-5 patients, which leads to a decreased stability of both proteins, as shown in patient lymphocytes. Activity of natural killer and cytotoxic T cells was markedly reduced or absent, as determined by CD107 degranulation. Our findings thus identify a key role for STXBP2 in lytic granule exocytosis. PMID- 19804850 TI - A common variation in EDAR is a genetic determinant of shovel-shaped incisors. AB - Shovel shape of upper incisors is a common characteristic in Asian and Native American populations but is rare or absent in African and European populations. Like other common dental traits, genetic polymorphisms involved in the tooth shoveling have not yet been clarified. In ectodysplasin A receptor (EDAR), where dysfunctional mutations cause hypohidrotic ectodermal dysplasia, there is a nonsynonymous-derived variant, 1540C (rs3827760), that has a geographic distribution similar to that of the tooth shoveling. This allele has been recently reported to be associated with Asian-specific hair thickness. We aimed to clarify whether EDAR 1540C is also associated with dental morphology. For this purpose, we measured crown diameters and tooth-shoveling grades and analyzed the correlations between the dental traits and EDAR genotypes in two Japanese populations, inhabitants around Tokyo and in Sakishima Islands. The number of EDAR 1540C alleles in an individual was strongly correlated with the tooth shoveling grade (p = 7.7 x 10(-10)). The effect of the allele was additive and explained 18.9% of the total variance in the shoveling grade, which corresponds to about one-fourth of the heritability of the trait reported previously. For data reduction of individual-level metric data, we applied a principal-component analysis, which yielded PC1-4, corresponding to four patterns of tooth size; this result implies that multiple factors are involved in dental morphology. The 1540C allele also significantly affected PC1 (p = 4.9 x 10(-3)), which denotes overall tooth size, and PC2 (p = 2.6 x 10(-3)), which denotes the ratio of mesiodistal diameter to buccolingual diameter. PMID- 19804853 TI - The mangled extremity. PMID- 19804854 TI - How to identify the asymptomatic high-risk patient? Foreword. PMID- 19804855 TI - How to identify the asymptomatic high-risk patient? AB - Worldwide, cardiovascular diseases are among the leading causes of mortality and morbidity with ever-increasing prevalence. Early targeted initiation of preventive measures would be of great benefit and can provide a major opportunity in reducing mortality and morbidity. To this end, accurate identification of individuals who are still asymptomatic but at elevated risk is essential. However, traditional risk assessment fails to recognize a substantial proportion of patients at high risk while a large proportion of individuals are classified as having intermediate risk, leaving management uncertain. Additional strategies to further refine risk assessment are therefore highly needed. To this end, the use of biomarkers and noninvasive imaging modalities has been proposed. The aim of this review is to provide an overview of the different approaches that are available or under development to improve the identification of asymptomatic individuals at elevated risk for cardiovascular diseases events. PMID- 19804856 TI - Using promoter replacement and selection for loss of heterozygosity to generate an industrially applicable sake yeast strain that homozygously overproduces isoamyl acetate. AB - By application of the high-efficiency loss of heterozygosity (HELOH) method for disrupting genes in diploid sake yeast (Kotaka et al., Appl. Microbiol. Biotechnol., 82, 387-395 (2009)), we constructed, from a heterozygous integrant, a homozygous diploid that overexpresses the alcohol acetyltransferase gene ATF2 from the SED1 promoter, without the need for sporulation and mating. Under the conditions of sake brewing, the homozygous integrant produced 1.4 times more isoamyl acetate than the parental, heterozygous strain. Furthermore, the homozygous integrant was more genetically stable than the heterozygous recombinant. Thus, the HELOH method can produce homozygous, recombinant sake yeast that is ready to be grown on an industrial scale using the well-established procedures of sake brewing. The HELOH method, therefore, facilitates genetic modification of this rarely sporulating diploid yeast strain while maintaining those characteristics required for industrial applications. PMID- 19804857 TI - Comparative analysis of a CFo ATP synthase subunit II homologue derived from marine and fresh-water algae. AB - Comparative analysis was performed with a CFo ATP synthase subunit II homologue (CFo-II) derived from marine or fresh-water algae. The marine algae-derived CFo II-transformed Escherichia coli grew and accumulated ATP more vigorously in NaCl or Cadmium containing medium, suggesting that this gene was useful for the development of stress-tolerant plant. PMID- 19804858 TI - Ester synthesis reaction with CALB displaying yeast whole cell biocatalyst: effect of organic solvent and initial water content. AB - The effect of the reaction conditions on the ester synthesis reaction with CALB displaying yeast whole cells was determined. Utilization of hydrophobic organic solvent improved the efficiency of the ester synthesis reaction. Also the initial water content was important for the expression of the ester synthesis activity of CALB displaying yeast whole cells. PMID- 19804859 TI - Lignocellulose degradation and enzyme production by Irpex lacteus CD2 during solid-state fermentation of corn stover. AB - The white rot fungus Irpex lacteus CD2 was incubated on corn stover under solid state fermentation conditions for different durations, from 5 days up to 120 days. Lignocellulose component loss, enzyme production and Fe3+-reducing activity were studied. The average weight loss ranged from 1.7% to 60.5% during the period of 5-120 days. In contrast to lignin, hemicellulose and cellulose were degraded during the initial time period. After 15 days, 63.0% of hemicellulose was degraded. Cellulose was degraded the most during the first 10 days, and 17.2% was degraded after 10 days. Lignin was significantly degraded and modified, with acid insoluble lignin loss being nearly 80% after 60 days. That weight loss, which was lower than the total component loss, indicated that not all of the lost lignocellulose was converted to carbon dioxide and water, which was indicated by the increase in soluble reducing sugars and acid soluble lignin. Filter paper activity, which corresponds to total cellulase activity, peaked at day 5 and remained at a high level from 40 to 60 days. High hemicellulase activity appeared after 30 days. No ligninases activity was detected during the incipient stage of lignin removal and only low lignin peroxidase activity was detected after 25 days. Apparently, neither of the enzymatic peaks coincided well with the highest amount of component loss. Fe3+-reducing activity could be detected during all the decay periods, which might play an important role in lignin biodegradation by I. lacteus CD2. PMID- 19804861 TI - Key role for transketolase activity in erythritol production by Trichosporonoides megachiliensis SN-G42. AB - Erythritol is an important sugar alcohol industrially produced only by fermentation. The highly osmophilic yeast-like fungi, Trichosporonoides megachiliensis SN-G42, enables commercial production of erythritol with a high conversion from glucose to erythritol of more than 47%. However, the microbial production pathway of erythritol remains unclear. In the present study, the activities of enzymes in the pentose phosphate pathway of Trichosporonoides megachiliensis SN-G42 used for industrial erythritol production were measured under various culture conditions to examine the production mechanism and the key enzymes. As a result, the various enzyme activities of this organism are revealed in the pentose phosphate pathway, i.e., those of hexokinase, glucose-6-phosphate dehydrogenase, gluconate dehydrogenase, transketolase, transaldolase, and erythrose reductase. In the cultures in which erythritol was produced after completion of cell growth, the enzyme activities of the pentose phosphate pathway were higher than those of the TCA cycle. In particular, transketolase activity was correlated with erythritol productivity under various production cultures with different agitation speeds and thiamine concentrations. These results suggest that erythritol may be produced mainly through the pentose phosphate pathway. In addition, the high activity of transketolase is required to produce abundant intermediates, which results in high erythritol productivity. As such, transketolase appears to be a key-enzyme for erythritol production in the organism studied. PMID- 19804860 TI - Kinetic modeling and sensitivity analysis of xylose metabolism in Lactococcus lactis IO-1. AB - We proposed a kinetic simulation model of xylose metabolism in Lactococcus lactis IO-1 that describes the dynamic behavior of metabolites using the simulator WinBEST-KIT. This model was developed by comparing the experimental time-course data of metabolites in batch cultures grown in media with initial xylose concentrations of 20.3-57.8 g/l with corresponding calculated data. By introducing the terms of substrate activation, substrate inhibition, and product inhibition, the revised model showed a squared correlation coefficient (r2) of 0.929 between the experimental time-course of metabolites and the calculated data. Thus, the revised model is assumed to be one of the best candidates for kinetic simulation describing the dynamic behavior of metabolites. Sensitivity analysis revealed that pyruvate flux distribution is important for higher lactate production. To confirm the validity of our kinetic model, the results of the sensitivity analysis were compared with enzyme activities observed during increasing lactate production by adding natural rubber serum powder to the xylose medium. The experimental results on pyruvate flux distribution were consistent with the prediction by sensitivity analysis. PMID- 19804862 TI - Acrylamide degradation by filamentous fungi used in food and beverage industries. AB - Filamentous fungi (24 strains) used in food and beverage industries were investigated for acrylamide-degradation ability: Aspergillus oryzae KBN1010 showed the highest ability. Little acrylic acid was produced but no glycidamide was detected during AA degradation in roasted green tea; therefore, A. oryzae could be used for reducing the AA concentration. PMID- 19804863 TI - Influence of the electron acceptor on nitrite reductase gene (nir) diversity in an activated sludge community. AB - Analyses of the nitrite reductase gene diversities (nirK and nirS) in an activated sludge community fed with both nitrite and glucose were conducted. Results suggest that the topology of nirK and nirS gene fragment-based phylogenetic trees is influenced more by the available electron acceptor than by the carbon source. A denitrification reactor was operated for 53 days and a clone library constructed when the denitrifying communities in the SBR were supplied with both nitrite and glucose. Half of the nirK and nearly all the nirS gene fragments formed a cluster that was separate from a cluster containing nirK and nirS sequences derived from other communities in nitrate-fed reactors. On the other hand, nirK and nirS fragments obtained with glucose as the carbon source were similar to those detected in communities fed with other carbon sources. PMID- 19804864 TI - Comparison of archaeal and bacterial community structures in heavily oil contaminated and pristine soils. AB - Archaeal and bacterial community structures in heavily oil-contaminated and pristine soils were compared using denaturing gradient gel electrophoresis and 16S rRNA gene libraries. The results showed that archaeal diversity was more complex in the contaminated soil than in the uncontaminated control soil. Archaeal populations in the contaminated soil consisted mainly of Euryarchaeota, with abundant methanogen-like operational taxonomic units (OTUs) and OTUs related to the phylogenetically diverse group, candidate division I, corresponding to rice cluster V. In contrast, only halophilic archaea were found in the pristine soil. Bacterial community structures also differed significantly between the contaminated and pristine soils. More clones from the contaminated soil were related to known hydrocarbon-degrading bacteria, implying that microorganisms with the potential to degrade petroleum were well-established. These results provide further insights into the composition of microbial communities in oil contaminated soils. PMID- 19804865 TI - Two-phased hyperthermophilic anaerobic co-digestion of waste activated sludge with kitchen garbage. AB - For co-digestion of waste activated sludge with kitchen garbage, hyperthermophilic digester systems that consisted of an acidogenic reactor operated at hyperthermophilic (70 degrees C) and a methanogenic reactor operated at mesophilic (35 degrees C), thermophilic (55 degrees C) or hyperthermophilic (65 degrees C) conditions in series were studied by comparing with a thermophilic digester system that consisted of thermophilic (55 degrees C) acidogenic and methanogenic reactors. Laboratory scale reactors were operated continuously fed with a substrate blend composed of concentrated waste activated sludge and artificial kitchen garbage. At the acidogenic reactor, solubilization efficiencies of chemical oxygen demand (COD), carbohydrate and protein at 70 degrees C were about 39%, 42% and 54%, respectively, and they were higher than those at 55 degrees C by around 10%. The system of acidogenesis at 70 degrees C and methanogenesis at 55 degrees C was stable and well-functioned in terms of treatment performances and low ammonium nitrogen concentrations. Microbial community analysis was conducted using a molecular biological method. The key microbe determined at the hyperthermophilic acidogenesis step was Coprothermobacter sp., which was possibly concerned with the degradation of protein in waste activated sludge. The present study proved that the hyperthermophilic system was advantageous for treating substrate blends containing high concentrations of waste activated sludge. PMID- 19804866 TI - Sensitive fluorescent microplate bioassay using recombinant Escherichia coli with multiple promoter-reporter units in tandem for detection of arsenic. AB - Genetically modified bacterial biosensors can detect specific environmental compounds. Here, we attempted to establish a fluorescent microplate method to detect arsenic using recombinant Escherichia coli cells transformed with plasmids harboring three tandem copies of the ars promoter/operator-the gene for green fluorescent protein (gfp). In the biosensors, one copy of arsR, whose transcription is autoregulated by the ars promoter/operator and ArsR in the genome of E. coli, was placed in trans in another plasmid under the control of isopropyl-1-thio-beta-D-galactopyranoside-inducible promoter. First, this manipulation enabled regulation of the arsR expression at an adequate level. Second, the copy number of reporter unit also affected signal and noise. When the plasmid harboring three copies of the reporter unit was used, the signal-to-noise ratio doubled and the detection limit decreased from 20 to 7.5 microg L(-1) As(III), compared to the use of the plasmid harboring one copy of the ars promoter/operator-arsR-gfp. Thus, segregation of arsR from the ars promoter/operator-gfp using two plasmids is effective in regulating the signal-to noise ratio and the detection limit with the different functions. PMID- 19804867 TI - Biodegradation of crystal violet by low molecular mass fraction secreted by fungus. AB - Biodegradation of crystal violet by white rot fungus Pleurotus ostreatus BP in rice straw solid-state medium mostly depended on low molecular mass fraction (LMMF) and the hypothetical metabolic pathways of crystal violet by LMMF was proposed by the biodegradation of crystal violet and by the deduced six metabolites formed. PMID- 19804868 TI - Analysis of interaction between liposome membranes induced by stress condition: utilization of liposomes immobilized on indium tin oxide electrode. AB - NBD-cholesterol (NBD-Ch)-modified liposome was immobilized on indium tin oxide (ITO) electrode via the covalent binding method. The transfer of NBD-Ch between the immobilized liposomes and the target liposomes was observed by using a fluorescent microscope. The addition of liposome suspension co-incubated with alpha-chymotrypsin or stimuli-responsive polymer to the surface of the above ITO electrode, enhanced the liposome-liposome interaction, resulting in the promotion of NBD-Ch transfer. The apparent transfer rate constant of NBD-Ch was found to be correlated with the index for the liposome-liposome interaction evaluated by an immobilized liposome chromatography. This suggests that the present method using the liposome-immobilized ITO electrode was effective to evaluate the liposome liposome interaction induced by the protein or the stimuli-responsive polymer under stress conditions. PMID- 19804869 TI - Lipid production in Porphyridium cruentum grown under different culture conditions. AB - Autotrophic growth of Porphyridium cruentum under 18:12 h and 12:12 h light:dark cycles showed the maximum cell concentration of 2.1 g-dry wt./L, whereas the specific growth rate, 0.042 (1/h), at 18:6 h is faster than that of 12:12 h, 0.031 (1/h), respectively. The highest lipid accumulation level, 19.3 (%, w/w), was achieved at 12:12 h cycle. Under dark cultivation condition with 10 g/L of glucose, the lipid accumulation in the cell was 10.9 (%, w/w), whereas the heterotrophic growth with glycerol as the carbon resource showed low level of cell concentration and lipid production, compared to that of glucose. The glucose was decided to be a suitable carbon resource for the heterotrophic growth of P. cruentum. The lipids from P. cruentum seemed be feasible for biodiesel production, because over 30% of the lipid was C16-C(18:1). The cultivation time and temperature were important factors to increase the maximum cell concentration. Extending the cultivation time helps maintain the maximum cell concentration, and higher lipid accumulation was achieved at 25 degrees C, compared to 35 degrees C. The fed-batch cultures showed that, under the light condition, the specific production rate was slightly decreased to 0.4% lipid/g dry wt./day at the later stage, whereas, under the dark condition, the specific production rate was maintained to be a maximum value of 1.1% lipid/g-dry wt./day, even in the later stage of cultivation. The results indicate that the heterotrophic or 12:12 h cyclic mixotrophic growth of P. cruentum could be used for the production of biodiesel in long-term fed-batch cultivation of P. cruentum. PMID- 19804870 TI - Effects of siRNAs targeting the spacer sequence of plant RNAi vectors on the specificity and efficiency of RNAi. AB - Small interfering RNAs (siRNAs) generated from the spacer region of hairpin RNAs were not detected in the RNA interference (RNAi) plants targeting the fatty acid desaturase gene. The expression of the desaturase gene was stably suppressed even when siRNAs targeting the spacer sequences were introduced into this plant. PMID- 19804871 TI - Novel reporter cell line to analyze cytokine-mediated expression regulation of c myc gene. AB - Growth-promoting cytokines induce expression of nuclear proto-oncogenes that play critical roles in the regulation of cell proliferation. c-myc gene is one of those nuclear proto-oncogenes, whose regulation mechanisms of cytokine-mediated expression are not fully understood. Here, I generated a green fluorescent protein (GFP) reporter system that faithfully reflects interleukin-3 (IL-3) induced c-myc gene expression. Flowcytometric analysis revealed cytokine-specific expression of reporter GFP. Kinetics of GFP mRNA expression was similar to that of endogenous c-myc mRNA. The reporter cell line will be a useful tool for studies of cell proliferation regulation through analysis of cytokine-induced c myc gene expression. PMID- 19804872 TI - Improved trimethoprim-resistance cassette for prokaryotic selections. AB - Many of the antibiotic resistance elements used in molecular biology have idiosyncratic limitations. For example, beta-lactam selections rely on antibiotics that are unstable to hydrolysis and allow satellite colonies to form upon extended incubation, and tetracycline selections typically give rise to widely varying colony sizes and lower transformation efficiencies. Although prokaryotic Type II dihydrofolate reductase (dfr) genes have long been considered to have potential utility for the selection of plasmids and mobile elements in bacteria, practical limitations to the quality of those selections, mostly relating to background and inefficiency, have led for the most part to their underuse. I describe here the construction of a Type IIa dfr prokaryotic expression cassette that confers strong resistance against trimethoprim (Tmp), a bactericidal dfr inhibiting antibiotic. The Tmp-resistance cassette provides consistent and efficient selections and plasmid transformation frequencies equivalent to those encountered with beta-lactamases. PMID- 19804873 TI - Insertional chromatin immunoprecipitation: a method for isolating specific genomic regions. AB - We established a novel method, insertional chromatin immunoprecipitation (iChIP), for isolation of specific genomic regions. In iChIP, specific genomic domains are immunoprecipitated with antibody against a tag, which is fused to the DNA-binding domain of an exogenous DNA-binding protein, whose recognition sequence is inserted into the genomic domains of interest. The iChIP method will be a useful tool for dissecting chromatin structure of genomic region of interest. PMID- 19804874 TI - Charged liposome affects the translation and folding steps of in vitro expression of green fluorescent protein. AB - The role of the charged liposome on the in vitro expression of green fluorescent protein (GFP) was investigated, focusing on its elemental steps such as transcription, translation and folding. The total GFP expression was enhanced to 145% when a neutral liposome (POPC: 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocoline) was added externally to a cell-free translation system. On the contrary, the addition of the charged liposome composed of POPC with anionic 1 palmitoyl-2-oleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (POPG) or cationic stearyl amine (SA) inhibited the total GFP expression, depending on the surface charge density of liposome. In transcription, the RNA synthesis was enhanced regardless of the variation of the surface charge, indicating that transcription was enhanced due to the stabilization of RNA structure by its hydrophobic interaction with liposome. Translation was inhibited by cationic liposome although it was enhanced by anionic liposome and neutral liposome. On the other hand, the folding was not inhibited in the presence of neutral liposome, whereas anionic liposome and cationic liposome inhibited the folding in proportion to the their surface charges, suggesting that the total GFP expression was controlled by a charged liposome in the translation step and folding step. PMID- 19804875 TI - Clinical implications of the obesity paradox in patients with coronary artery disease. PMID- 19804876 TI - Common molecular causes for congenital heart defects and microcephaly. PMID- 19804878 TI - The effect of digital cervical examination on group B streptococcal culture. AB - OBJECTIVE: We sought to evaluate whether group B streptococcus (GBS) detection is altered by the digital cervical examination. STUDY DESIGN: A total of 302 women undergoing the clinical GBS culture had a digital cervical examination and a repeated GBS culture. Statistical comparison of pre-post culture results were performed with kappa and McNemar tests. RESULTS: The clinical prevalence of GBS was 19.5%. Discordant results were seen in 30/302 (9.9%) paired cultures (kappa = 0.68; 95% confidence interval, 0.568-0.783). An initially negative GBS culture result was positive on repeated testing in 13/243 (5.3%) pairs. Initially positive cultures were negative on repeated testing in 17/59 (28.8%) pairs. Patients with discordant results had similar characteristics as the remainder of the study group. Given the observed proportion of discordant results (9.9%), the study had 80% power to detect a 5% difference between discordant pairs. CONCLUSION: Paired GBS cultures showed a good level of agreement. The 28.8% rate of positive cultures becoming negative is clinically concerning and warrants further study. PMID- 19804879 TI - Structural characterization of a heteropolysaccharide isolated from hot water extract of the stems of Amaranthus tricolor Linn. (Amaranthus gangeticus L.). AB - A water-soluble polysaccharide (PS-I), isolated from the aqueous extract of the stems of Amaranthus tricolor Linn. (Amaranthus gangeticus L.), was found to consist of L-arabinose, methyl-D-galacturonate, D-galactose, and 3-O-Ac-L rhamnose in a molar ratio of nearly 1:1:1:1. On the basis of total acid hydrolysis, methylation analysis, periodate oxidation, and NMR studies ((1)H, (13)C, TOCSY, DQF-COSY, NOESY, ROESY, HMQC, and HMBC), the structure of the repeating unit of the PS-I is determined as: -->5)-alpha-L-Araf-(1-->4)-beta-D Galp-(1-->2)-3-O-Ac-beta-L-Rhap-(1-->4)-alpha-D-Gal-A6Me-(1--> PMID- 19804880 TI - Synthesis of N-tetra-O-acetyl-beta-D-glucopyranosyl-N'-(4',6'-diarylpyrimidin-2' yl)thioureas. AB - Some 2-amino-4,6-diarylpyrimidines 2 have been prepared from substituted benzylideneacetophenones and guanidine hydrochloride in the presence of alkali by conventional heating in alcoholic medium and microwave heating in solvent-free conditions. N-(2,3,4,6-Tetra-O-acetyl-beta-D-glucopyranosyl)-N'-(4',6' diarylpyrimidin-2'-yl)thioureas 4 have been synthesized by reaction of per-O acetylated glucopyranosyl isothiocyanate 1 and substituted 2-amino-4,6 diarylpyrimidines 2. Two different methods have been used, namely, refluxing in anhydrous dioxane and solvent-free microwave-assisted coupling. The second procedure afforded higher yields in much shorter reaction times. The compounds 2 and 4 were tested for their antibacterial and antifungal activities in vitro against Staphylococcus epidermidis, Enterobacter aerogenes and Candida albicans by disc diffusion method. PMID- 19804881 TI - Structure of the cell wall polysaccharides of probiotic bifidobacteria Bifidobacteriumbifidum BIM B-465. AB - The composition and structure of cell wall polysaccharides of a biotechnologically promising probiotic strain of bifidobacteria Bifidobacteriumbifidum BIM B-465 were established by monosaccharide and methylation analyses along with 2D (1)H-(1)H homonuclear and (1)H-(13)C heteronuclear correlation NMR spectroscopy. The major polysaccharide represents a branched glucogalactan consisting of heptasaccharide repeating units having the following structure: -->6)-alpha-D-Glcp-(1-->3)-beta-D-Galf-(1-->3)-alpha-D-Glcp (1-->2)-beta-D-Galf-(1-->3)-alpha-D-Galp-(1-->3)-alpha-D-Glcp-(1--> [structure: see the text]. The second isolated polysaccharide is a branched glucan with the main chain of (1-->6)-linked alpha-d-glucopyranose residues, approximately 60% of which are 2-substituted with a single alpha-d-glucopyranosyl group. This polysaccharide is a characteristic of various microorganisms, including some lactobacteria studied earlier. PMID- 19804882 TI - Nurses' attitudes to mental illness: a comparison of a sample of nurses from five European countries. AB - BACKGROUND: Mental health problems are of serious concern across Europe. A major barrier to the realisation of good mental health and well-being is stigma and discrimination. To date there is limited knowledge or understanding of mental health nurses' attitudes towards mental illness and individuals experiencing mental health problems. OBJECTIVES: To describe and compare attitudes towards mental illness and those experiencing mental health problems across a sample of registered nurses working in mental health settings from five European countries and the factors associated with these attitudes. DESIGN: A questionnaire survey. SETTINGS: A total of 72 inpatient wards and units and five community facilities in Finland, Lithuania, Ireland, Italy and Portugal. PARTICIPANTS: 810 registered nurses working in mental health settings. METHODS: The data were collected using The Community Attitudes towards the Mentally Ill (CAMI) scale, which is a 40-item self-report questionnaire. The data were analysed using quantitative methods. RESULTS: Nurses' attitudes were mainly positive. Attitudes differed across countries, with Portuguese nurses' attitudes being significantly more positive and Lithuanian nurses' attitudes being significantly more negative than others'. Positive attitudes were associated with being female and having a senior position. CONCLUSIONS: Though European mental health nurses' attitudes to mental illness and people with mental health problems differ significantly across some countries, they are largely similar. The differences observed could be related to wider social, cultural and organisational circumstances of nursing practice. PMID- 19804883 TI - Increased intima thickness of the radial artery in individuals with prehypertension and hypertension. AB - BACKGROUND: We have used a novel ultra high-frequency (55MHz) ultrasound technique to non-invasively measure the radial arterial vessel wall and separate the intima-media (IMT) complex into measurements of intima and media thickness (IT and MT). Since no previous study has measured IT and MT separately in individuals with prehypertension and hypertension, the aim of the current study was to measure IT and MT thickness of the radial arteries among individuals with prehypertension, hypertension and healthy subjects. METHODS AND RESULTS: Individuals with prehypertension (n=32), hypertension (n=34) and healthy subjects (n=29) underwent ultra high-resolution ultrasound of the radial artery. Individuals with prehypertension showed a 14% increase in IT compared to healthy subjects (0.083+/-0.020mm versus 0.073+/-0.015mm; p<0.05), whereas no difference was seen in MT. Individuals with hypertension showed a 12% increase of in IT compared to healthy subjects (0.082+/-0.018mm versus 0.073+/-0.015mm, p<0.05), whereas no differences were seen regarding MT. Prehypertensive and hypertensive individuals did not differ regarding IT and MT. CONCLUSION: Both prehypertension and hypertension are associated with thickening of the intimal layer of the radial artery. The present data indicates that intima thickening appears early during the development of hypertension even when blood pressure is only slightly elevated. PMID- 19804884 TI - Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) and risk of cardiovascular disease in older adults: results from the Cardiovascular Health Study. AB - OBJECTIVE: To examine associations between lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) antigen level (mass) and enzymatic activity (activity) and cardiovascular disease (CVD) in older adults. METHODS: We examined associations of Lp-PLA(2) mass and activity with incident myocardial infarction (MI; n=508), stroke (n=565) and CVD death (n=665) using Cox regressions adjusted for age, sex, ethnicity and CVD risk factors in 3949 older adults, aged > or =65 years at baseline, from the Cardiovascular Health Study (CHS). RESULTS: Lp-PLA(2) was associated with incident CVD events in these older adults. Hazard ratios (95% confidence intervals) for highest versus lowest tertiles of Lp-PLA(2) mass were 1.49 (1.19-1.85) for MI, 1.21 (0.98-1.49) for stroke and 1.11 (0.92-1.33) for CVD death. The highest tertile of Lp-PLA(2) activity was associated with MI (1.36; 1.09-1.70) and CVD death (1.23; 1.02-1.50). Combined Lp-PLA(2) tertile 3 and CRP>3mg/l, compared to Lp-PLA(2) tertile 1 and CRP<1mg/l, was associated with MI (2.29; 1.49-3.52) for Lp-PLA(2) mass and MI (1.66; 1.10-2.51) and CVD death (1.57; 1.08-2.26) for activity. For MI, both mass and activity added excess risk to elevated CRP alone ( approximately 20% excess risk) and activity added excess risk for CVD death ( approximately 12%). CONCLUSION: Lp-PLA(2) mass and activity were associated with incident CVD events in older adults in CHS. Lp-PLA(2) and CRP were independent and additive in prediction of events. While associations were modest, these results support further exploration of Lp-PLA(2) to identify older individuals at risk for CVD. PMID- 19804886 TI - In the middle of it all: mutual mechanical regulation between the nucleus and the cytoskeleton. AB - The nucleus is typically treated as the large phase-dense or easy-to-label structure at the center of the cell which is manipulated by the governing mechanical machinery inside the cytoplasm. However, recent evidence has suggested that the mechanical properties of the nucleus are important to cell fate. We will discuss many aspects of the structural and functional interconnections between nuclear mechanics and cellular mechanics in this review. There are numerous implications for the progression of many disease states associated with both nuclear structural proteins and cancers. The nucleus itself is a large organelle taking up significant volume within the cell, and most studies agree that nuclei are significantly stiffer than the surrounding cytoplasm. Thus when a cell is exposed to force, the nucleus is exposed to and helps resist that force. The nucleus and nucleoskeleton are interconnected with the cellular cytoskeleton, and these connections may aid in helping disperse forces within tissues and/or with mechanotransduction. During translocation and transmigration the nucleus can act as a resistive element. Understanding the role of mechanical regulation of the nucleus may aid in understanding cellular motility and crawling through confined geometries. Thus the nucleus plays a role in developing mechanical territories and niches, affecting rates of wound healing and allowing cells to transmigrate through tissues for developmental, repair or pathological means. PMID- 19804887 TI - Dispersive liquid-liquid-liquid microextraction combined with liquid chromatography for the determination of chlorophenoxy acid herbicides in aqueous samples. AB - A novel sample preparation method "Dispersive liquid-liquid-liquid microextraction" (DLLLME) was developed in this study. DLLLME was combined with liquid chromatography system to determine chlorophenoxy acid herbicide in aqueous samples. DLLLME is a rapid and environmentally friendly sample pretreatment method. In this study, 25microL of 1,1,2,2-tetrachloroethane was added to the sample solution and the targeted analytes were extracted from the donor phase by manually shaking for 90s. The organic phase was separated from the donor phase by centrifugation and was transferred into an insert. Acceptor phase was added to this insert. The analytes were then back-extracted into the acceptor phase by mixing the organic and acceptor phases by pumping those two solutions with a syringe plunger. After centrifugation, the organic phase was settled and removed with a microsyringe. The acceptor phase was injected into the UPLC system by auto sampler. Fine droplets were formed by shaking and pumping with the syringe plunger in DLLLME. The large interfacial area provided good extraction efficiency and shortened the extraction time needed. Conventional LLLME requires an extraction time of 40-60min; an extraction time of approximately 2min is sufficient with DLLLME. The DLLLME technique shows good linearity (r(2)>or=0.999), good repeatability (RSD: 4.0-12.2% for tap water; 5.7-8.5% for river water) and high sensitivity (LODs: 0.10-0.60microg/L for tap water; 0.11 0.95microg/L for river water). PMID- 19804885 TI - Recent advances and new opportunities in lung mechanobiology. AB - Lung function is inextricably linked to mechanics. On short timescales every breath generates dynamic cycles of cell and matrix stretch, along with convection of fluids in the airways and vasculature. Perturbations such airway smooth muscle shortening or surfactant dysfunction rapidly alter respiratory mechanics, with profound influence on lung function. On longer timescales, lung development, maturation, and remodeling all strongly depend on cues from the mechanical environment. Thus mechanics has long played a central role in our developing understanding of lung biology and respiratory physiology. This concise review focuses on progress over the past 5 years in elucidating the molecular origins of lung mechanical behavior, and the cellular signaling events triggered by mechanical perturbations that contribute to lung development, homeostasis, and injury. Special emphasis is placed on the tools and approaches opening new avenues for investigation of lung behavior at integrative cellular and molecular scales. We conclude with a brief summary of selected opportunities and challenges that lie ahead for the lung mechanobiology research community. PMID- 19804889 TI - Activation of microglia/macrophages expressing phosphorylated S6 ribosomal protein in a case of hemimegalencephaly with progressive calcification and atrophy. AB - A 3-year-old boy with right hemimegalencephaly (HME) showed massive calcification in the subcortical white matter and progressive atrophy of the affected hemisphere. Hemispherotomy was successful in amelioration of the patient's intractable epilepsy, and a surgical specimen from the epileptic focus was examined pathologically. Disarrangement of cortical layers along with dysmorphic appearance of neurons, balloon cells in the cortex and white matter, bi-layered calcifications in the superficial cortical layer and subcortical white matter, heterotopic neurons in the white matter, and diffuse astrogliosis were noted. Perivascular clustering of alpha-B-crystallin positive balloon cells was occasionally observed in the area of calcification. A diffuse increase was observed in the number of CD68-positive microglia/macrophages, particularly in perivascular and peri-calcification areas. These cells were often located within the calcification foci, which implicates their participation in the calcification process. Phosphorylated S6 ribosomal protein (P-S6) was expressed in large-sized neurons and numerous balloon cells, as well as in CD68-positive cells. In contrast, phosphorylated mammalian target of rapamycin (mTOR) was expressed in a small percentage of astrocytes, and phosphorylated p70S6 kinase was rarely identified in perivascular cells. These findings suggest that inflammatory processes have contributed to the pathogenesis of progressive calcification and atrophy in the megalencephalic hemisphere in this patient. Dissociation of expression of mTOR cascade components is common to other reported cases of HME, but P-S6 expression in microglia/macrophages has not been recognized. The cellular mechanism and significance of P-S6-specific activation of the mTOR cascade in HME, particularly in the inflammatory cell lineage, should be explored further. PMID- 19804890 TI - Validation of the Parkinson's disease sleep scale in Japanese patients: a comparison study using the Pittsburgh Sleep Quality Index, the Epworth Sleepiness Scale and Polysomnography. AB - OBJECTIVE: Characteristic sleep disturbance in patients with Parkinson's disease (PD) was evaluated using a subjective questionnaire called the PD sleep scale (PDSS). In this study we sought to examine the relationship between the results from the PDSS with those from the Pittsburgh Sleep Quality Index (PSQI), the Epworth Sleepiness Scale (ESS), and polysomnography (PSG) in Japanese PD patients. SUBJECTS AND METHODS: Subjects were 79 PD patients and 79 age and gender matched controls. The length of morbidity in patients with PD was 8.4+/ 8.0 years, and their Hoehn and Yahr grade was 2.9+/-1.0. All subjects completed the PDSS, PSQI, and ESS questionnaires, and we compared the results from patients with PD to controls. We also evaluated the correlation among the PDSS, PSQI, and ESS in patients with PD. Moreover, we performed PSG on 33 of 79 PD patients, and examined the correlation between the PDSS and PSG. RESULTS: PDSS total scores and subscales from patients with PD were significantly lower than those in controls, except for items 3, 8, and 14. PDSS scores had significant internal consistency and significant correlation with PSQI and ESS scores. The total PDSS score also correlated with sleep efficiency as measured by PSG. In subscales of the PDSS, night psychosis was negatively correlated with percentage of REM sleep without atonia on PSG. DISCUSSION: Based on our comparisons with PSQI, ESS, and PSG, the PDSS appears to be a reliable tool to evaluate the characteristics of sleep disturbances in PD patients. PMID- 19804891 TI - [Biomarkers in risk stratification of acute coronary syndrome without ST elevation: is there anything after troponin?]. PMID- 19804892 TI - Glazed (vision) and confused. AB - A 60-year-old man presented with vitritis and optic neuropathy in the setting of headaches and behavioral changes. MRI brain revealed bilateral temporal lobe inflammation consistent with limbic encephalitis. He was subsequently diagnosed with small cell lung cancer with a paraneoplastic syndrome characterized by CRMP5 IgG as a cause of his symptoms. His visual symptoms improved markedly after anti inflammatory therapy and his cognitive symptoms were mildly better following systemic chemotherapy. The clinical presentation, pathophysiology, and therapy of CRMP5 associated paraneoplastic syndromes are discussed. PMID- 19804893 TI - Biotreatment and bioassessment of heavy metal removal by sulphate reducing bacteria in fixed bed reactors. AB - In this work a batch-optimised mixture (w/w %: 6% leaves, 9% compost, 3% Fe(0), 30% silica sand, 30% perlite, 22% limestone) was investigated in a continuous fixed bed column reactor for the treatment of synthetic acid-mine drainage (AMD). A column reactor was inoculated with sulphate-reducing bacteria and fed with a solution containing sulphate and heavy metals (As(V), Cd, Cr(VI), Cu and Zn). At steady state, sulphate abatement was 50+/-10%, while metals were totally removed. A degradation rate constant (k) of 0.015+/-0.001h(-1) for sulphate removal was determined from column data by assuming a first order degradation rate. Reduction of AMD toxicity was assessed by using the nematode Caenorhabditis elegans as a test organism. A lethality assay was performed with the toxicants before and after the treatment, showing that only 5% of the animals were still alive after 48h in presence of the contaminants, while the percentage increased to 73% when the nematodes were exposed to the solution eluted from the column. PMID- 19804894 TI - Phosphorylated hydroxyethylamines as novel inhibitors of the bacterial cell wall biosynthesis enzymes MurC to MurF. AB - Enzymes involved in the biosynthesis of bacterial peptidoglycan represent important targets for development of new antibacterial drugs. Among them, Mur ligases (MurC to MurF) catalyze the formation of the final cytoplasmic precursor UDP-N-acetylmuramyl-pentapeptide from UDP-N-acetylmuramic acid. We present the design, synthesis and biological evaluation of a series of phosphorylated hydroxyethylamines as new type of small-molecule inhibitors of Mur ligases. We show that the phosphate group attached to the hydroxyl moiety of the hydroxyethylamine core is essential for good inhibitory activity. The IC(50) values of these inhibitors were in the micromolar range, which makes them a promising starting point for the development of multiple inhibitors of Mur ligases as potential antibacterial agents. In addition, 1-(4 methoxyphenylsulfonamido)-3-morpholinopropan-2-yl dihydrogen phosphate 7a was discovered as one of the best inhibitors of MurE described so far. PMID- 19804895 TI - Effect of triclosan and triclocarban biocides on biodegradation of estrogens in soils. AB - We have investigated the effect of antimicrobials triclosan (TCS) and triclocarban (TCC) on biodegradation of 17beta-Estradiol (E2) and 17 alpha Ethynylestradiol (EE2) in a sandy soil from South Australia. Two separate batch studies were conducted. In the first, the rates of loss of E2 and EE2 were determined at time intervals of 0, 3, 7, 14, 21, 28 and 56 d after initial spiking of soil with each estrogen at 1 mg kg(-1) and the antimicrobials at 10 and 100 mg kg(-1). Little loss of E2 and EE2 (<15%) under sterile conditions was noted compared to rapid loss in non-sterile soil (>60% in 24h). There were no measurable effects on estrogen degradation by the two antimicrobials at spiked concentrations up to 100 mg kg(-1). The experiments were repeated to study degradation rates of the estrogens within the first 24h (0, 3, 8, 24h), 3 d and then weekly to 56 d. Again, E2 and EE2 degradation was not significantly affected by the presence of TCS up to 100 mg kg(-1) (p>0.05). However TCS did significantly affect biodegradation of the estrogens when the soils were spiked with 1000 mg kg(-1) of TCS (p<0.0005). In contrast, presence of TCC in soil showed no significant effect on biodegradation of the two compounds up to 1000 mg kg(-1) (p>0.05). Considering environmental concentrations of the antimicrobials reported in the literature, it is highly unlikely these biocides would have any adverse impact on biodegradation of E2 or EE2 in soils. PMID- 19804896 TI - The effects of ethylene glycol and ethanol on the body mass and elemental composition of insects collected with pitfall traps. AB - Insects often used as accumulation indicators of hazardous elements. Pitfall traps with ethylene glycol as trapping fluid are frequently used to collect insects. We studied the effect of glycol and preservation with ethanol on the elemental composition of hand collected firebugs. Control samples were stored in a freezer and the following treatments were used: insect kept in trapping fluid for 2 weeks, and for 1 month, trapping fluid for 2 weeks plus 2 weeks in ethanol, and trapping fluid for 1 month plus ethanol for 1 month. Insects kept in trapping fluid gained mass with respect to control: 26% for the short trapping and 37% for the long trapping. Preservation in ethanol reversed this effect in each case. Trapping fluid did not alter the dry mass. A significant loss in dry mass only occurred in the long trapping plus long preservation treatment. We analysed the concentration of eight elements: Ca, Cu, K, Mg, Mn, Na, Sr and Zn. We found significant difference in the concentrations of elements among the four treatments in the case of all elements, except magnesium and zinc. Our results indicate the potential of both certain trapping fluids as well as preservation in ethanol influencing the concentration of certain elements in insects. Live trapping for collection and storage in under freezing conditions for preservation could be a more reliable method if quantitative analytical studies are to be performed, when invertebrates are used as indicators of the presence and concentrations of hazardous substances in the environment. PMID- 19804897 TI - Organochlorine pesticides and polybrominated diphenyl ethers in irrigated soils of Beijing, China: levels, inventory and fate. AB - Limited information on the levels, inventory and fate of Organochlorine pesticides (OCPs) and Polybrominated diphenyl ethers (PBDEs) in the soils irrigated by sewage or wastewater is available. In this study, variation in concentrations, profiles and fate of OCPs and PBDEs were investigated using soil samples collected from a region irrigated by sewage, mixed water and clean water in the east of Beijing, China. No significant variation was observed among groups, except for penta-BDEs. The measured SigmaOCPs and SigmaPBDEs residues ranged from 6.4 to 171.2 ng g(-1) (dw) and 501.9 to 3310.7 pg g(-1) (dw), respectively. SigmaDDTs and BDE-209 were the most abundant congeners accounting for about 76% of SigmaOCPs and 93% of SigmaPBDEs. Concentrations of hexachlorocyclohexanes (HCHs), dichlorodiphenyltrichloroethane (DDTs) and its major degradation products, and hexachlorobenzene (HCB) ranged from 1.2 to 11.4 ng g(-1) (dw), 4.0 to 155.6 ng g(-1) (dw) and 0.3 to 3.4 ng g(-1) (dw), respectively. The major DDT degradation products were p,p'-DDT and p,p'-DDE. The major hexachlorocyclohexane (HCH) isomer in irrigated soils is beta-HCH, reflecting its higher affinity to solids and resistance to degradation than other isomers. Both alpha-HCH/beta-HCH and p,p'-DDT/p,p'-DDE ratios were log-normally distributed and negatively correlated to log(SigmaHCHs) and log(SigmaDDTs), respectively, suggesting no significant recent application of OCPs. Individual BDE congeners, SigmaPBDEs and SigmaOCPs were significantly correlated with total organic carbon (TOC). Moreover, a good correlationship between SigmaPBDEs and black carbon (BC) was obtained but not between SigmaOCPs and BC. Sewage irrigation did not have obvious effect on their contaminant levels and inventory of OCPs and PBDEs. PMID- 19804898 TI - Spatial structure of trace elements in extensive biomonitoring surveys with terrestrial mosses. AB - The size of the area affected by contamination processes mainly depends on the type of contaminant and the way it is emitted. In regular sampling, the size of the sampling grid will therefore interact with the scale at which the contamination processes are produced, for each contaminant, so that the grid will only enable characterization of those processes that occur at scales larger than the grid, i.e. large scale processes. The present study analysed the data corresponding to tissue concentrations of Cd, Cu, Fe, Hg, Mn, Pb, Se, V and Zn in the terrestrial moss Pseudoscleropodium purum obtained in regular sampling surveys with grids of different sizes (15x15, 7.5x7.5 km) plus a series of accessory points located at 1 km from the 15x15 km sampling grid; all sampling sites were located in Galicia and were sampled in March 2004. The objective of the study was to characterize the interaction between the scale of the sampling grid and the scale at which the contamination processes are produced, for each contaminant, to enable adjustment of the size of the grids to the scale at which the contamination processes are mainly produced. For this purpose, the spatial structure of the concentration of each element was analysed by use of semivariograms created with a robust estimator of the semivariance. The results of the study verified the existence of a real interaction between the scale of the sampling grid and the scale of the contamination processes. The results also demonstrated how in the study area, the contamination processes associated with Cd, Pb and V are generally small scale processes, whereas those associated with Cu, Hg and Se are generally large scale processes. PMID- 19804899 TI - Activated protein C stimulates osteoblast proliferation via endothelial protein C receptor. AB - INTRODUCTION: Bone is continually remodeled by the action of osteoblasts, osteocytes, and osteoclasts. Resting osteoblasts are able to proliferate and differentiate into mature osteoblasts when physiologically required, as after tissue injury. Activated protein C (APC) is a serine protease that functions in anticoagulation, anti-inflammation, anti-apoptosis, cell proliferation, and wound repair. In this study, we examined the effect of APC on osteoblast proliferation and differentiation. MATERIALS AND METHODS: We examined the presence of protein C in human fracture hematoma by immunohistochemical staining. We then evaluated the effect of APC, diisopropyl fluorophosphate-inactivated APC (DIP-APC) or protein C zymogen on normal human osteoblast (NHOst) proliferation using tetrazolium salt assay in the presence or absence of aprotinin, hirudin, protein C, antibody against protein C, endothelial protein C receptor (EPCR) or protease-activated receptor (PAR)-1. Finally, activation of p44/42 MAP kinase was evaluated by Western blot analysis. RESULTS: Both APC and DIP-APC increased osteoblast proliferation in a dose-dependent manner, while protein C did not. The APC induced increased proliferation of osteoblast was not affected by aprotinin, hirudin, and anti-protein C antibody which inhibits the protease activity of APC. Treatment with protein C or anti-EPCR antibody which inhibits APC binding to EPCR inhibited APC-mediated osteoblast proliferation, while treatment with anti-PAR-1 antibody did not. APC promoted the phosphorylation of p44/42 MAP kinase within osteoblasts; this effect was inhibited by the anti-EPCR antibody. CONCLUSIONS: APC stimulates osteoblast proliferation by activating p44/42 MAP kinase through a mechanism that requires EPCR but not PAR-1 or the proteolytic activity of APC. APC generated at fracture sites may contribute to fracture healing by promoting osteoblast proliferation. PMID- 19804900 TI - Induction of death receptor ligand-mediated apoptosis in epithelial ovarian carcinoma: The search for sensitizing agents. AB - OBJECTIVE: To assess the abilities of cisplatin, paclitaxel, and flexible heteroarotinoid (Flex-Het) compound (SHetA2) to sensitize ovarian cancer cells to induction of the extrinsic apoptosis pathway by death receptor ligands, tumor necrosis factor alpha (TNFalpha), and TNF-related apoptosis-inducing ligand (TRAIL). STUDY DESIGN: The effects of various combinations of TNFalpha, TRAIL, cisplatin, paclitaxel, and SHetA2 on viability and apoptosis in two established ovarian cancer cell lines, A2780 and SK-OV-3, and normal human primary endometrial cultures were measured with a cytotoxicity assay, flow cytometric analysis of annexin-V, and propidium iodide staining and Western blot analysis of caspase 8 and 3 activation. RESULTS: Ovarian cancer and normal cells were resistant to TNFalpha and TRAIL. Cisplatin and paclitaxel did not increase sensitivity to these agents in either cell type. In contrast, combination of SHetA2 with TNFalpha or TRAIL induced a synergistic induction of apoptosis in cancer cells that involved activation of the extrinsic pathway caspase 8 and executioner caspase 3. The TRAIL combination was more potent than the TNFalpha combination. SHetA2 did not harm the viability of normal cells as a single agent or in combination with the death receptor ligands. CONCLUSIONS: SHetA2, but not cisplatin or paclitaxel, can overcome resistance of ovarian cancer cells to TNFalpha and TRAIL without increasing sensitivity of normal cells to these death receptor ligands. PMID- 19804901 TI - Sustained progression-free survival with weekly paclitaxel and bevacizumab in recurrent ovarian cancer. AB - OBJECTIVE: To determine efficacy, toxicity, and survival in patients with recurrent epithelial ovarian cancer (EOC) receiving combination of weekly paclitaxel and biweekly bevacizumab (PB). METHODS: We reviewed chemotherapy logs identifying all patients receiving combination PB. Toxicities were graded using CTCAEv3.0 criteria. Response rates (RR) were measured using RECIST criteria or by CA-125 levels per modified Rustin criteria. RR and progression-free survival (PFS) were determined and plotted using Kaplan-Meier survival analysis. RESULTS: Fifty-one patients receiving at least two cycles of chemotherapy were evaluable for survival and 55 patients receiving one cycle of PB were evaluable in toxicity analysis. The mean number of previous regimens was four. The overall median PFS was 7 months and median OS was 12 months. The overall response rate (ORR) was 60% (CR 25% and PR 35%). Median PFS for complete and partial responders were 14 and 5 months respectively. Stable disease was seen in 26% with median PFS of 6 months. Thirteen experienced treatment delays for a variety of factors. The most G3/4 toxicities were fatigue (16%), hematologic (9%) and neurotoxicity (7%). Three patients (5%) experienced bowel perforations. CONCLUSIONS: Combination of paclitaxel and bevacizumab is feasible and demonstrates an acceptable toxicity profile and a high response rate. These observations should be useful in planning future clinical trials with this combination therapy. PMID- 19804903 TI - The ART approach using glass-ionomers in relation to global oral health care. AB - Dental caries is the most prevalent non-communicable disease in the world. Its management in high-income countries over the last four decades has resulted in relatively low caries prevalence in child and adolescent populations. In low- and middle-income countries, caries management is virtually non-existent and this may lead to serious physical and mental complications, particularly in children. Toothache is predominantly treated by extracting the cavitated tooth. Absence of restorative oral care is partly due to the copying from high-income countries, of restorative treatment reliant on electrically driven equipment and often inappropriate for use in many low- and middle-income countries. Atraumatic Restorative Treatment (ART), which does not rely on electrically driven equipment, has yielded good results over the last two decades. ART uses hand instruments and high-viscosity glass-ionomers. Its introduction into public oral healthcare systems has been piloted in several countries. Initial short-term results show that the introduction of ART, using high-viscosity glass-ionomers, has increased the ratio of restorations to extractions. Moreover, the percentage of ART restorations in relation to the total number of restorations placed increased steeply after its introduction and has remained high. However, ART introduction faced a few barriers, the most important being high patient workloads and the absence of a constant supply of dental instruments and glass ionomers. High-viscosity glass-ionomer has become an essential element in public oral healthcare systems, particularly in those operating inadequately. PMID- 19804902 TI - A phase II evaluation of pemetrexed (Alimta, LY231514, IND #40061) in the treatment of recurrent or persistent endometrial carcinoma: a phase II study of the Gynecologic Oncology. AB - OBJECTIVE: To estimate the anti-tumor activity of pemetrexed in patients with advanced or recurrent carcinoma of the endometrium and to determine the nature and degree of toxicity. METHODS: A multicenter phase II trial was conducted by the Gynecologic Oncology Group (GOG). Patients must have had advanced or recurrent measurable carcinoma of the endometrium and failed one prior chemotherapy regimen. Pemetrexed at a dose of 900 mg/m(2) was administered as an IV infusion over 10 min every 21 days. RESULTS: From May 1, 2006 to July 31, 2007, 27 patients were entered by 10 member institutions of the GOG with two patients being deemed ineligible. A total of 101 cycles were administered with 28% of patients receiving five or more cycles. Overall, the treatment was well tolerated. More serious toxicities (grade 3 and 4) included anemia in 20%, leukopenia in 40%, neutropenia in 48%, and constitutional in 16%. No treatment related deaths were reported. One patient (4%) had a partial response. Eleven patients (44%) had stable disease and eleven (44%) patients had increasing disease. Response could not be assessed in two patients (7%). Median progression free survival was 2.7 months and overall survival was 9.4 months. CONCLUSION: Pemetrexed has minimal activity in the treatment of recurrent or persistent endometrial carcinoma at the dose and schedule tested. PMID- 19804904 TI - Clonal hematopoiesis in Philadelphia chromosome-negative bone marrow cells of chronic myeloid leukemia patients receiving dasatinib. AB - A clonal cytogenetic abnormality was observed in Philadelphia chromosome-negative bone marrow cells of 6/27 chronic myeloid leukemia patients (+8 in 4, -7 in 1, and 20q- in 1) with dasatinib-induced remissions. The X-linked human androgen receptor gene assay demonstrated clonality in one additional patient. Single nucleotide polymorphism array analysis revealed somatic uniparental disomy involving chromosome 17(p12-pter) in another patient. The TP53 gene had a 5' splice site deletion of exon 6 that caused alternative splicing, frame shifting and introduction of a premature stop codon. After three years, no patient developed myelodysplastic syndrome or acute myeloid leukemia. PMID- 19804905 TI - Factors discriminating among profiles of resilience and psychopathology in children exposed to intimate partner violence (IPV). AB - OBJECTIVE: To evaluate the social and emotional adjustment of 219 children in families with varying levels of intimate partner violence (IPV) using a model of risk and protection. To explore factors that differentiate children with poor adjustment from those with resilience. METHODOLOGY: Mothers who experienced IPV in the past year and their children ages 6-12 were interviewed. Standardized measures assessed family violence, parenting, family functioning, maternal mental health, and children's adjustment and beliefs. RESULTS: Using cluster analysis, all cases with valid data on the Child Behavior Checklist, Child Depression Inventory, General Self-Worth and Social Self-Competence measures were described by four profiles of children's adjustment: Severe Adjustment Problems (24%); children who were Struggling (45%); those with Depression Only (11%); and Resilient (20%) with high competence and low adjustment problems. Multinomial logistic regression analyses showed children in the Severe Problems cluster witnessed more family violence and had mothers higher in depression and trauma symptoms than other children. Resilient and Struggling children had mothers with better parenting, more family strengths and no past violent partner. Parents of children with Severe Problems were lacking these attributes. The Depressed profile children witnessed less violence but had greater fears and worries about mother's safety. CONCLUSION: Factors related to the child, to the mother and to the family distinguish different profiles of adjustment for children exposed to IPV who are living in the community. Resilient children have less violence exposure, fewer fears and worries, and mothers with better mental health and parenting skills, suggesting avenues for intervention with this population. PRACTICE IMPLICATIONS: Findings suggest that child adjustment is largely influenced by parent functioning. Thus, services should be targeted at both the child and the parent. Clinical interventions shaped to the unique needs of the child might also be tested with this population. PMID- 19804906 TI - Intimate partner violence and immigration laws in Canada: how far have we come? AB - Immigrant women face numerous, and sometimes insurmountable, barriers in reporting and seeking services for intimate partner violence (IPV). A number of these obstacles relate to immigration laws, policies and legal processes they encounter due to their immigration status and sponsorship relationship. The present study was conducted in Canada, in an urban centre that boasts one of the largest immigrant populations in the world. Using a focus group methodology within a participatory action research framework, this investigation sought to identify factors that facilitate or impede women from coming forward and disclosing IPV, and traced their help-seeking actions. Qualitative data from helping professionals and women reveal that in cases of sponsorship breakdown due to IPV, the criteria required for a viable immigration application are unrealistic, and in many cases impossible to meet in situations of domestic abuse. These data indicate that despite claims to the contrary, laws and policies related to immigration have remained stable for over a decade. Systemic and structural barriers that these create for abused women are still clearly present in immigration laws and policies. The result is that many women stay in abusive relationships, often with their children, for prolonged periods of time accruing serious negative mental health effects. Implications are discussed to help inform policy and practice. PMID- 19804907 TI - Development of the hsp110-heparanase vaccine to enhance antitumor immunity using the chaperoning properties of hsp110. AB - The purpose of the study is to identify the novel approach of hsp110-heparanase vaccine in virtue of the chaperoning properties of hsp110, and to explore the antitumor potential. To test it, the recombinant hsp110 and heparanase proteins form into complex vaccine during heat shock, and the antitumor immunity is observed. We demonstrate that hsp110 and heparanase can form into complex vaccine, and the vaccine can enhance the specific IFN-gamma production and cytotoxic T cell response. Additionally, immunization of the vaccine can also significantly suppress the established tumor growth and prolong the life span of tumor-bearing mice. Thus, the use of the recombinant hsp110 and heparanase proteins to form into complex vaccine represents a novel and promising way for the therapeutical strategy of tumor. PMID- 19804908 TI - Endosomal damage and TLR2 mediated inflammasome activation by alkane particles in the generation of aseptic osteolysis. AB - Ultra-high molecular weight polyethylene is widely used as a bearing surface in prosthetic arthroplasty. Over time the generation of implant-derived wear particles can initiate an inflammatory reaction characterized by periprosthetic inflammation and ultimately bone resorption at the prosthetic bone interface. Herein we present evidence that the different sized particles as well as the different length alkane polymers generated by implant wear leads to a two component inflammatory response. Polymeric alkane structures, with side chain oxidations, directly bind and activate the TLR-1/2 signaling pathway. Whereas micron- and nanometer-sized particulate debris are extensively phagocyted and induce enlargement, fusion and disruption of endosomal compartments. The resulting lysosomal damage and subsequent enzymatic leakage induces the NALP3 inflammasome activation as determined by cathepsins S and B cytosolic release, Caspase 1 activation and processing of pro-IL-1beta, and pro-IL-18. These two processes synergistically results in the initiation of a strong inflammatory response with consequent cellular necrosis and extracellular matrix degradation. PMID- 19804909 TI - The human neonatal B cell response to respiratory syncytial virus uses a biased antibody variable gene repertoire that lacks somatic mutations. AB - The human Ab repertoire exhibits restrictions during fetal life characterized by biases of variable gene segment usage and lack of junctional diversity. We tested the hypotheses that Ab repertoire restriction persists in the early postnatal period and contributes to the observed poor quality of specific Ab responses made by neonates to viruses and vaccines. We analyzed the molecular determinants of B cell responses in humans to respiratory syncytial virus (RSV). Analysis of the variable gene segment usage of adult RSV-specific B cells revealed a repertoire profile in these cells similar to that seen in randomly selected B cells, which was V(H)3-dominant. Four gene segments (V(H)3-23, V(H)3-30, V(H)3-33 and V(H)4 04) accounted for almost half of the V(H) genes used. In contrast, very young infant RSV-specific antibodies exhibited a biased repertoire characterized by comparable use of the V(H)1, V(H)3, and V(H)4 families, and less common use of the four immunodominant gene segments. Infants and children older than three months used an antibody repertoire similar to that of adults. Mutational analysis revealed that the antibody variable genes of infants under three months of age also possessed significantly fewer somatic mutations in both framework and complementarity-determining region (CDR) regions than those of adults, even in a child with recurrent RSV infection. These data suggest that neonates use a biased antibody gene repertoire that is less V(H)3-focused and that possesses a dramatically lower frequency of somatic mutations. These biased features of the RSV-specific repertoire likely contribute to the poor functional Ab response in very young infants. PMID- 19804910 TI - Hepatitis B virus sensitizes hepatocytes to complement-dependent cytotoxicity through downregulating CD59. AB - Hepatitis B virus (HBV) infection afflicts over 350 million people worldwide and is a leading cause of hepatitis, cirrhosis and hepatocellular carcinoma. HBV replicates noncytopathically in hepatocytes, and most of the hepatic injury is caused by the immune response to the virus. While most studies focused on the adaptive immune response, the role of the innate immune response, especially the complement activation, in HBV infection remains obscure. To identify proteins that are involved in the pathogenesis of HBV infection, we carried out gene microarray analysis to compare the gene expression profile of HBV transgenic BALB/c mice with that of control mice. CD59 mRNA, which encodes an important complement regulatory protein (CRP) expressed on cell surface, was found to be significantly downregulated in HBV transgenic liver, a result that was further confirmed by RT-PCR and real-time PCR. To explore the relationship between CD59 and HBV infection, we examined the effect of HBV on CD59 expression and complement-dependent cytolysis in two hepatocyte cell lines. We found that HBV could significantly downregulate CD59 expression and sensitize cells to complement-dependent lysis. Blocking CD59 function using a CD59-specific antibody greatly diminished the HBV effect. Similar CD59 downregulation was also observed in the livers of patients with chronic HBV infection. These results demonstrate that HBV can sensitize hepatocytes to complement-dependent cytotoxicity (CDC) through downregulating CD59, which may lead to the activation of complement system and cause liver inflammation. PMID- 19804911 TI - Prevalence and correlates of depressive disorders in elderly with type 2 diabetes in primary health care settings. AB - OBJECTIVE: Depression is associated with poor glycemic control and complications in people with type 2 diabetes. We assessed the prevalence of depressive symptoms and antidepressant medication use among elderly with and without type 2 diabetes and the association between depression and diabetes complications. RESEARCH DESIGN AND METHODS: In 2004-2006, the Primary Health Care research in Type 2 Diabetes Study applied the Beck Depression Inventory II (BDI-II) to 458 participants with type 2 diabetes (47% male, aged 65 + or -8.9 years, type 2 diabetes duration 19 + or - 8.7 years) and 546 participants without diabetes (non diabetic group) (51% male, aged 59 + or - 8.7 years). Use of antidepressant medication was self-reported. Depressive disorder was defined as a BDI-II score >14 and/or use of antidepressant medication. Occurrence of diabetes complications (retinopathy, blindness, neuropathy, diabetes-related amputation, and kidney or pancreas transplantation) was self-reported. RESULTS: Mean BDI-II score, adjusted for age and sex, was significantly higher in participants with type 2 diabetes than in non diabetic participants (least-squares mean + or - SE: 7.4 + or - 0.3 vs. 5.0 + or - 0.3; P<0.0001). The prevalence of depressive disorder (as defined by BDI-II>14 and/or antidepressant use) in participants with type 2 diabetes was significantly higher than that of age- and sex-adjusted non diabetic participants (32.1 vs. 16.0%, P<0.0001). Type 2 diabetic participants reported using more antidepressant medications (20.7 vs. 12.1%, P = 0.0003). More type 2 diabetic than non diabetic participants were classified as depressed by BDI-II cut score (17.5 vs. 5.7%, P<0.0001) or by either BDI-II cut score or antidepressant use (32.1 vs. 16.0%, P<0.0001). Participants reporting diabetes complications (n = 209) had higher mean BDI-II scores than those without complications (10.7 + or - 9.3 vs. 6.4 + or - 6.3, P<0.0001). CONCLUSIONS: Depression is highly prevalent in type 2 diabetes and requires further study on assessment and treatment. PMID- 19804912 TI - Methodological issues in the obsessive-compulsive spectrum. AB - The obsessive-compulsive spectrum is a heterogeneous class of conditions. Recently, expert consensus has emerged regarding possible candidate disorders [Mataix-Cols, D., Petrusa, A., Leckman, J.F., 2007. Issues for DSM-V: How should obsessive-compulsive and related disorders should be classified. American Journal of Psychiatry, 164, 1313-1314]. Further, expert survey data suggest that obsessive-compulsive disorder is composed of overlapping subtypes. However, methodological approaches for testing whether candidate disorders properly belong in the spectrum have varied widely, and do not necessarily differentiate conditions from subtypes. We describe preliminary methodological and statistical recommendations for a systematic approach to determining what constitutes a subtype, how to determine inclusion in the spectrum, and means for ruling out candidate disorders. PMID- 19804913 TI - AUY954, a selective S1P(1) modulator, prevents experimental autoimmune neuritis. AB - Experimental autoimmune neuritis (EAN) is a T cell-mediated autoimmune inflammatory demyelinating disease of the peripheral nervous system and an animal model of human inflammatory demyelinating polyradiculoneuropathy. AUY954, which targets selectively the sphingosine-1-phosphate receptor 1 (S1P(1)), is known to sequester lymphocytes into secondary lymphoid tissues. In EAN rats, AUY954 greatly prevented paraparesis if administrated from the day of immunization. T cell, B cell, and macrophage infiltration, inflammatory demyelination, and local expression of interleukine-17 and matrix metalloproteinase-9 in sciatic nerves of EAN rats were significantly decreased by AUY954 treatment. Therefore, S1P(1) modulation might be a potential treatment option for inflammatory neuropathies. PMID- 19804914 TI - Auditory function in women with autoimmune inner ear diseases and their offspring. AB - OBJECTIVE: The precise cause of congenital sensorineural hearing loss (CSNHL) is unclear in many cases. In a previous study we found that offspring from guinea pigs with autoimmune sensorineural hearing loss (ASNHL) exhibited signs of SNHL. Here we studied women with autoimmune inner ear diseases (AIED) and their offspring. Our aim was to determine if autoimmune damage may be one of the causes of CSNHL. METHODS: Thirty-eight pregnant women with AIED were recruited. Thirty three had ASNHL; one with autoimmune delayed endolymphatic hydrops (ADEH) and four with autoimmune Meniere's disease (AIMD). The following were assessed in all women: audiogram, auditory brain stem response (ABR), otoacoustic emission (OAE), vestibular function test and presence of inner ear antigens. The following were assessed in offspring from these women: OAE, ABR and presence of inner ear antigens. RESULTS: Five of the 38 children born to women with AIED had SNHL (an incidence much higher than normal). OAEs were not inducible in these children shortly after birth or within 46-100 days after birth. Abnormal ABR findings were apparent in these five children and inner ear antigens were detected in three of the five children (the mother's of these children were also positive for inner ear antigens). CONCLUSIONS: These preliminary findings suggest that the prevalence of congenital ASNHL may be increased in offspring born to women with AIED. PMID- 19804915 TI - Bacteremic methicillin-resistant Staphylococcus aureus deep neck abscess in a newborn--case report and review of literature. AB - We describe an unusual localization of methicillin-resistant Staphylococcus aureus infection (MRSA) in a very young newborn. A 3-week-old male infant was admitted with fever, irritability, sialorrhea and stiffed left neck. The ENT examination revealed a deep neck mass and an ultrasound examination showed diffuse swelling of the left latero-pharyngeal area. A CT examination confirmed a deep neck abscess with difficult-to-define borders. Blood and nasopharyngeal cultures returned positive for MRSA. Treatment was started with intravenous teicoplanin and continued for 14 days with a marked decrease in abscess's dimensions and improvement in patient's general condition. MRSA should be suspected in the etiology and treatment of neck abscesses in newborns, infants and young children. PMID- 19804916 TI - Preventive Veterinary Medicine-What has changed in 2009? PMID- 19804919 TI - Regional trends in multidrug-resistant infections in German intensive care units: a real-time model for epidemiological monitoring and analysis. AB - A new surveillance module for multidrug-resistant (MDR) bacteria was added to the intensive care component of the German Nosocomial Infection Surveillance System. Participating intensive care units (ICUs) report data on all patients colonised or infected with meticillin-resistant Staphylococcus aureus (MRSA), vancomycin resistant enterococci (VRE) and extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae (ESBL-EC/KP). To determine the regional distribution of MRSA, VRE and ESBL-EC/KP in Germany, incidence densities (IDs) of these bacteria per 1000 patient-days were calculated for each ICU and pooled for ICUs of five German regions for the years 2005 and 2006. A total of 176 ICUs including 284 142 patients and 1 021 579 patient-days reported data concerning 5490 cases of MDR bacteria. The pooled IDs were 4.54, 0.54 and 0.29 cases per 1000 patient-days for MRSA, ESBL-EC/KP and VRE cases and 1.56, 0.32 and 0.13 per 1000 patient-days for MRSA, ESBL-EC/KP and VRE infections, respectively. While there were no significant differences in the incidence densities of MRSA infections between regions, the IDs of VRE and ESBL-EC/KP infections showed significant regional variation. The regions also differed in the proportion of ICUs per region that reported at least one infection with MRSA, ESBL-EC/KP or VRE in 2005-2006 and these differences ranged from 82% to 91% for MRSA, from 34% to 76% for ESBL-EC/KP and from 8% to 42% for VRE. This new surveillance module enables ICUs to monitor the occurrence of MDR bacteria by comparing local incidence densities with a national reference and shows significant regional variation of MDR bacteria in Germany. PMID- 19804918 TI - Differential localization to cytoplasm, nucleus or P-bodies of yeast PKA subunits under different growth conditions. AB - Our aim in this work was to further characterize the complexity and specificity of the three different isoforms (Tpk1, Tpk2 and Tpk3) of the catalytic and regulatory (Bcy1) subunits of PKA in Saccharomyces cerevisiae. We thus analyzed the subcellular localization of the PKA subunits in living cells by using strains carrying GFP (green fluorescent protein) fused to each PKA subunit. During exponential growth on glucose, both Bcy1 and Tpk2 localized in the nucleus, whereas Tpk1 and Tpk3 showed a mixed pattern of nucleo-cytoplasmic localization. During exponential growth on glycerol and during stationary phase, the PKA subunits showed mostly cytoplasmic localization, with the peculiarity that Tpk2 and Tpk3 but not Bcy1, were found associated to P-bodies and EGP bodies. Tpk3 was accumulated into P-bodies during glucose deprivation and hyper osmotic stress. Deletion of Tpk3 altered the kinetics of P-body formation. Analysis of protein expression showed that the relative expression pattern of each Tpk changes from low levels under fermentative metabolism to higher levels during stationary phase. The increase in Tpk levels produced an imbalance with Bcy1 levels. Our data suggest that the signaling specificity through PKA in yeast could be mediated by a particular subcellular localization of each isoform of Tpk. PMID- 19804921 TI - [Ethical and legal problems in severe dementia. The right to die in peace]. AB - The progression toward a natural death, without medical intervention, which was the normal route until fairly recently, could be attributed to the lack of means and medical facilities, influencing the development of events toward a normal death. The advances in medical technology that prolong life have created a false notion in our society that a longer life goes hand in hand with a high quality of life, which in most cases is false. The fact that intellectual and physical functions have sometimes been maintained or only minimally impaired has served to create the obligation to try all medical techniques, provoking clearly futile treatments and behaviors. In the terminally ill, our approach should be proportional to the needs of the patient and should be based on the principles of bioethics. A wise and not too academic bioethical approach should be able to help us to take the right decisions aimed at satisfying the patient's needs at the end of life. A bioethical approach that studies life not simply from the biological but also from the biographical perspective--with maintenance of life as a right rather than as an obligation--is required. PMID- 19804920 TI - [Treatment optimization in chronic hepatitis C virus infection]. AB - The treatment duration that obtains the optimal risk-benefit ratio in chronic hepatitis C infection is guided by viral kinetic data in weeks 4 and 12. Rapid virological response (RVR) and early virological response (EVR) have high positive and negative predictive value, respectively. Patients with genotype-1, RVR, without significant fibrosis and low baseline viral load (<600,000UI/ml) can receive treatment for 24 weeks without loss of efficacy, while the absence of EVR in these patients is a criterion for treatment interruption. Data on prolonging treatment to 72 weeks in patients with genotype 1 and a decrease of >2log in viremia without negativization of viremia in week 12 are contractictory. In patients with genotypes 2 and 3, 24-week treatment is superior to 16-week treatment, although 16-week treatment can be evaluated in patients with genotype 3 and RVR. In patients with genotype 2 and RVR, rates of RVR in 14-week treatment are similar to those in 24-week treatment, while in patients without RVR, treatment should be continued to 24 weeks. Key factors in treatment optimization are the weight-adjusted dose of ribavirin and therapeutic adherence. PMID- 19804922 TI - Professional collaboration in students of Medicine Faculty and School of Nursing. AB - This descriptive study has been planned to analyze the professional collaboration among the students of Ege University, Faculty of Medicine and School of Nursing. The study group consisted of 137 5th and 6th grade students from the Faculty of Medicine and 142 3rd and 4th grade students from the School of Nursing. The participation rate is 94%. For data collection, a questionnaire form (30 questions) which was specially developed for the purpose of the study and the Jefferson Scale of Attitudes toward Physician-Nurse Collaboration (15 questions) were used. Permission was granted for the research by Ege University School of Nursing Ethical Committee and the Office of the Dean of the Faculty of Medicine. The professional collaboration mean score of the students from the Faculty of Medicine who participated in the study was 30.40+/-5.82 and the professional collaboration mean score of the students from the School of Nursing was 26.11+/ 5.27. The difference between the mean scores of professional collaboration was found to be significant in terms of their profession (p<0.01). Physicians expressed more positive attitudes toward collaboration than nurses while female physicians expressed more positive attitudes toward collaboration than male physicians. PMID- 19804923 TI - Depositional behaviors of plutonium and thorium isotopes at Tsukuba and Mt. Haruna in Japan indicate the sources of atmospheric dust. AB - Monthly plutonium and thorium depositions at Tsukuba (28m asl) and Mt. Haruna (1370m asl) were measured during 2006 and 2007 (Jan 2006-Dec 2007 at Tsukuba, Nov 2006-Dec 2007 at Mt. Haruna). The monthly (239,240)Pu depositions ranged from 0.044 to 2.67mBq m(-2) at Tsukuba and from 0.05 to 0.9mBq m(-2) at Mt. Haruna during the measurement periods. Monthly (239,240)Pu deposition did not differ markedly between the two sites except in April 2007. Seasonal pattern of monthly (239,240)Pu depositions at both sites showed high in spring and low in summer, and typical of seasonal variations in northeastern Asia. Thorium deposition at Tsukuba was higher than that at Mt. Haruna except in May and June 2007. (230)Th/(232)Th activity ratios were used to partition deposition samples into locally and remotely derived fractions. The results revealed that a major proportion of total (239,240)Pu and Th deposits are derived from remote sources, especially in spring. PMID- 19804924 TI - Quantitative use of photography in orthognathic outcome assessment. AB - This study reports an independent audit of two aspects of orthognathic surgery, namely control of inter-alar width and mandibular outline asymmetry. Measurements were taken from standardized photographs of a consecutive series of 27 patients, using an on-screen digitizing program (IPTool). All patients had undergone bimaxillary osteotomies involving maxillary impaction and/or advancement, by one surgeon, using a cinch suture for nasal width control. Nine-twelve months after surgery, inter-alar width had increased by just 0.08 cm mean (SD 0.3). Four patients showed an increase of just over 2mm, whilst six showed a small reduction. Based on ratios of size (area) and shape (compactness) of the right and left mandibular segments, there was a small overall improvement in mandibular symmetry (0.019 and 0.005 respectively). Whilst in most of the patients the need for surgery was primarily the correction of antero-posterior and vertical discrepancies, five patients with demonstrable asymmetry showed a clear improvement. In three patients whose asymmetry scores were very mild pre treatment, there was a small, measured increase in asymmetry, but not to a degree that would be clinically noticeable. At a time when 3D imaging is still unavailable to many clinicians, the results of this study suggest that appropriate measurements taken from carefully standardized conventional photographs can provide a valid and objective means of assessing treatment outcome. PMID- 19804925 TI - Actinic granuloma affecting the upper lip: a rare and challenging clinical entity. AB - Actinic granuloma is a rare dermatological condition that can be difficult to diagnose, and the opinion of specialist dermatologists and histopathologists might be needed to confirm diagnosis. Our patient's lesion was not diagnosed until 3 years after initial presentation. We are aware of only 18 reported cases since 1982, and to our knowledge this is the first reported to affect the upper lip. The condition is slow to improve, and the most effective treatment remains unclear. We used intralesional steroid injections, but a small area has yet to resolve 9 years after presentation. PMID- 19804926 TI - Seasonal acclimation of the moss Polytrichum juniperinum Hedw. to natural and enhanced ultraviolet radiation. AB - Short- and long-term changes in the methanol-extractable UV-absorbing compounds and biomass of the pioneer moss Polytrichum juniperinum in response to natural and enhanced UV radiation were studied. Under natural conditions, the compounds were found to fluctuate seasonally. In summer these compounds correlated negatively with irradiation. The concentration was low in July after a period of simultaneous heat, drought and high irradiation. Transient positive correlation between daily concentration and UV was seen in June. The concentration increased towards autumn and was relatively high under snow. Two enhanced UV experiments were performed. Seasonality in the compounds was again observed, with negative correlations with irradiation. During the first weeks, a transient inhibition of compound production was observed after the daily UV-B treatment. After six years of modulated UV-treatment in situ, photosynthesizing biomass decreased under UV-B and increased under UV-A. A larger variation in the UV-absorbing compounds was observed under UV-B treatment. PMID- 19804927 TI - [Incidence and characteristics of asthma exacerbations in Barcelona (ASMAB II)]. AB - INTRODUCTION: A study has been made on the incidence and clinical characteristics of asthma exacerbations (AE) seen in hospital emergency departments (HED) and domiciliary services (DS) in Barcelona. PATIENTS AND METHODS: AEs were identified over a 56 day period during the months of October and November 2003 and the patients seen in university hospitals were interviewed. RESULTS: A total of 262 AE were identified, 188 in hospital emergency departments and 82 in domiciliary, which was a mean of 4.6 AE/day, giving an incidence of 0.37 AA/10(5) inhabitants. This incidence was no different from that obtained in the same months in 2002 (P>0.05). The mean age (+/-standard deviation) of the 143 cases of AE seen in university hospitals was 41+/-17 years; 87/143 (60%) were women and 57/138 (41%), smokers. In 36/61 (59%) the AE started in the home; in 88/132 (61%) it began 24 hours before arriving at the hospital centre. In 78/123 cases (63%) slow onset of nasal catarrh was noted before the AE and a viral infection was suspected. Inhaled corticoids were given to 45/112 patients (40%), and only one added oral corticoids 12 hours before admission. The AE were mild in 38/75 cases (51%), moderate in 28/75 (37%), severe in 8/75 (11%) and almost fatal in one patient (1.3%). Around 10% (13/137) of patients (10%) were re-admitted. CONCLUSIONS: The daily incidence of AE is 0.37/10(5) inhabitants. More than half of the exacerbations started after nasal catarrh and 11% of the AE were severe. PMID- 19804928 TI - Lipid resuscitation. PMID- 19804929 TI - Body temperature changes are associated with outcomes following in-hospital cardiac arrest and return of spontaneous circulation. AB - INTRODUCTION: Spontaneous changes in body temperature after return of circulation (ROSC) from cardiac arrest are common, but the association of these changes with outcomes in hospitalized patients who survive to 24h post-ROSC is not known. We tested the hypothesis that adults who experience temperature lability in the first 24h have worse outcomes compared with those who maintain normothermia. MATERIALS AND METHODS: A prospective observational study from a multicenter registry of cardiac arrests (National Registry of Cardiopulmonary Resuscitation) from 355 US and Canadian hospitals. 14,729 adults with return of circulation from a pulseless cardiac arrest. We excluded those who died or were discharged before 24h post-event, those made Do-Not-Resuscitate (DNR) within 24h of event, those that had a preceding trauma, and those with multiple cardiac arrests. Finally, we included only subjects that had both a lowest (T(min)) and highest (T(max)) body temperature value recorded during the first 24-h after ROSC, resulting in a study sample of 3426 patients. RESULTS: After adjustment for potential covariates, there was a lower odds of survival in those having an episode of hypothermia (adjusted odds ratio [OR], 0.62; 95% confidence interval [CI], 0.48-0.80), those having an episode of hyperthermia (OR, 0.67; 95% CI, 0.48-0.80), and those having an episode of both (OR, 0.59; 95% CI, 0.39-0.91). Among those who survived to discharge, there was also a lower odds of favorable neurologic performance in those who had an episode of hyperthermia (OR, 0.71; 95% CI, 0.51-0.98). CONCLUSIONS: Episodes of temperature lability following in-hospital resuscitation from cardiac arrest are associated with lower odds of surviving to discharge. Hyperthermia is also associated with fewer patients leaving the hospital with favorable neurologic performance. Further studies should identify whether therapeutic control over changes in body temperature after in-hospital cardiac arrest improves outcomes. PMID- 19804930 TI - Is external jugular vein cannulation feasible in emergency care? A randomised study in open heart surgery patients. AB - The optimal intravenous catheterisation site for emergencies is unknown. The external jugular vein might be preferable route compared to cubital veins in emergencies due to more rapid circulation time to heart and faster cardiac responses. However, the feasibility of the different venous catheterisation sites has not been compared in relation to catheterisation time and success rate. METHODS: We examined the time differences and success rates of external jugular compared to antecubital vein catheterisations. 32 paramedics and 28 emergency department residents performed external jugular and antecubital venous catheterisations on anesthetized patients scheduled for elective cardiac surgery. The primary outcome was catheterisation time and the secondary outcomes the failure rate and catheterisation times needed to succeed. RESULTS: Antecubital venous catheterisation was faster (113+/-89s) compared to external jugular vein catheterisation (156+/-112s), p=0.008 and the success rate was higher (93% compared to 68%, respectively, p=0.001). Less attempts were needed for antecubital vein catheterisations compared to external jugular vein catheterisations (p=0.002). For the antecubital vein, subjects needed two attempts in 6 patients and three attempts in 6 patients. For the external jugular vein, subjects needed two attempts in 13 patients and three attempts in 20 patients. Two (6%) paramedics and two (7%) residents failed to catheterise the antecubital vein. Nine (28%) paramedics and 10 (36%) residents failed to catheterise the external jugular vein. CONCLUSIONS: Antecubital vein catheterisation was faster and had a superior success rate compared to external jugular vein catheterisation. PMID- 19804931 TI - The first documented cardiac arrest rhythm in hospitalized patients with heart failure. AB - BACKGROUND: Patients with heart failure (HF) have abnormal cellular anatomy and myocardial mechanics that may impact the initial rhythm and subsequent outcomes in cardiac arrest (CA). HYPOTHESIS: Patients with pre-existing HF are less likely to have ventricular fibrillation/pulseless ventricular tachycardia (VF/pVT) as the first documented rhythm in CA and have poorer survival than patients without pre-existing HF. PURPOSE: Identify the first documented cardiac arrest rhythm (FDR) in hospitalized patients with and without a pre-existing history of HF. METHODS: We evaluated 60,389 consecutive, adult, index, pulseless CA events with documented initial rhythm in the National Registry of Cardiopulmonary Resuscitation. The primary endpoint was the FDR in patients with and without a history of pre-existing HF. Secondary endpoints were return of spontaneous circulation (ROSC), survival to discharge, and neurological outcome. RESULTS: Thirty three percent of patients had a pre-existing diagnosis of HF. HF patients were more likely to have VF/pVT (25.9 vs. 23.2%) and less likely to have asystole (34.4 vs. 35.3%, p=<.0001) than non-HF. There was no difference in survival to discharge (18.3 vs. 18.2%, p=.66), or good neurological outcomes (82.2 vs. 83.2%, p=.23) between the groups. Women were less likely to have VF/pVT as the first documented rhythm in both HF and non-HF groups. CONCLUSIONS: Hospitalized patients with HF are more likely than those without HF to have VF/pVT as the FDR in CA, however the clinical magnitude of this difference is small. Overall survival and neurological outcomes are no different than hospitalized arrest patients without HF. PMID- 19804932 TI - Predictors of resuscitation outcome in a swine model of VF cardiac arrest: A comparison of VF duration, presence of acute myocardial infarction and VF waveform. AB - INTRODUCTION: Factors that affect resuscitation to a perfusing rhythm (ROSC) following ventricular fibrillation (VF) include untreated VF duration, acute myocardial infarction (AMI), and possibly factors reflected in the VF waveform. We hypothesized that resuscitation of VF to ROSC within 3min is predicted by the VF waveform, independent of untreated VF duration or presence of acute MI. METHODS: AMI was induced by the occlusion of the left anterior descending coronary artery. VF was induced in normal (N=30) and AMI swine (N=30). Animals were resuscitated after untreated VF of brief (2min) or prolonged (8min) duration. VF waveform was analyzed before the first shock to compute the amplitude-spectral area (AMSA) and slope. RESULTS: Unadjusted predictors of ROSC within 3min included untreated VF duration (8min vs 2min; OR 0.11, 95%CI 0.02 0.54), AMI (AMI vs normal; OR 0.11, 95%CI 0.02-0.54), AMSA (highest to lowest tertile; OR 15.5, 95%CI 1.7-140), and slope (highest to lowest tertile; OR 12.7, 95%CI 1.4-114). On multivariate regression, untreated VF duration (P=0.011) and AMI (P=0.003) predicted ROSC within 3min. Among secondary outcome variables, favorable neurological status at 24h was only predicted by VF duration (OR 0.22, 95% CI 0.05-0.92). CONCLUSIONS: In this swine model of VF, untreated VF duration and AMI were independent predictors of ROSC following VF cardiac arrest. AMSA and slope predicted ROSC when VF duration or the presence of AMI were unknown. Importantly, the initial treatment of choice for short duration VF is defibrillation regardless of VF waveform. PMID- 19804933 TI - Induction and maintenance of in vivo ventricular fibrillation in rabbits. AB - AIM: To provide new sustainable in vivo models of ventricular fibrillation in rabbits. METHODS: New Zealand rabbits were submitted to anaesthesia and mechanical ventilation, after which ventricular fibrillation was induced through electrical stimulation (for 2min at 100Hz, with 2-ms pulses, 10mA, and 10V) directly to the heart. To that end, the animals were divided into two groups: right ventricle (n=11) and left ventricle (n=11). In group right ventricle, the thoracic cavity was exposed, and a catheter was introduced into the right ventricle via the right jugular vein. In group left ventricle, the thorax remained closed, and the catheter was introduced into the left ventricle via the left common carotid artery (cervical access). RESULTS: Sustained ventricular fibrillation was achieved in 100% of group right ventricle rabbits (n=11) and in 82% of group left ventricle rabbits (n=9). CONCLUSION: Both models proved appropriate for achieving sustained ventricular fibrillation. However, in view of the invasiveness of the procedure adopted in group right ventricle, the experimental conditions used in group left ventricle seemed more physiological and more effective in inducing sustained ventricular fibrillation. PMID- 19804934 TI - A population-based study on the prevalence and determinants of cardiopulmonary resuscitation in the last month of life for Taiwanese cancer decedents, 2001 2006. AB - BACKGROUND: The success rate of cardiopulmonary resuscitation (CPR) for cancer patients following in-hospital cardiac arrest has remained poor over the last 3 decades, but little is known about determinants of undergoing CPR for these patients at the end of life. OBJECTIVE: To determine the prevalence of CPR for Taiwanese cancer patients in the last month of life and the association between their undergoing CPR and patient demographics, disease characteristics, physician specialty, hospital characteristics, and availability of healthcare resources at the hospital and regional levels. METHODS: This retrospective cohort study examined administrative data for a cohort of 204,850 cancer decedents in 2001 2006. RESULTS: Rates of CPR decreased substantially over the study period, from 13.18% to 8.63%, and the adjusted odds ratio of undergoing CPR decreased significantly by a factor of 0.93 for each successive year. Taiwanese cancer patients were predisposed to undergo CPR in their last month of life if they were male, young, and unmarried (except for widowhood); had high comorbidity; had certain cancers (hematological malignancies, head and neck, esophageal, and prostate cancers); had a localized or newly diagnosed (within 1-2 months of death) cancer; had a non-oncologist as their primary physician; and received care at a non-teaching hospital. CONCLUSION: One-tenth of Taiwanese cancer patients underwent CPR in the last month of life, and the rates of CPR decreased substantially from 2001 to 2006. The propensity for CPR was influenced by patient demographics, disease characteristics, physician specialty, and teaching status of the patient's primary hospital. PMID- 19804935 TI - Intraosseous access in osteogenesis imperfecta (IO in OI). PMID- 19804936 TI - Techniques for artefact filtering from chest compression corrupted ECG signals: good, but not enough. PMID- 19804937 TI - Validation of a computational platform for the analysis of the physiologic mechanisms of a human experimental model of hemorrhage. AB - Computational models of integrative physiology may serve as a framework for understanding the complex adaptive responses essential for homeostasis in critical illness and resuscitation and may provide insights for design of diagnostics and therapeutics. In this study a computer model of human physiology was compared to results obtained from experiments using Lower Body Negative Pressure (LBNP) analog model of human hemorrhage. LBNP has been demonstrated to produce physiologic changes in humans consistent with hemorrhage. The computer model contains over 4000 parameters that describe the detailed integration of physiology based upon basic physical principles and established biologic interactions. The LBNP protocol consisted of a 5min rest period (0mmHg) followed by 5min of chamber decompression of the lower body to -15, -30, -45, and -60mmHg and additional increments of -10mmHg every 5min until the onset of hemodynamic decompensation (n=20). Physiologic parameters recorded include mean arterial pressure (MAP), cardiac output (CO), and venous oxygen saturation (SVO(2); from peripheral venous blood), during the last 30s at each LBNP level. The computer model analytic procedure recreates the investigational protocol for a virtual individual in an In Silico environment. After baseline normalization, the model predicted measurements for MAP, CO, and SVO(2) were compared to those observed through the entire range of LBNP. Differences were evaluated using standard statistical performance error measurements (median performance error (PE) <5%). The simulation results closely tracked the average changes observed during LBNP. The predicted MAP fell outside the standard error measurement for the experimental data at only LBNP -30mmHg while CO was more variable. The predicted SVO(2) fell outside the standard error measurement for the experimental data only during the post-LBNP recovery point. However, the statistical median PE measurement was found to be within the 5% objective error measure (1.3% for MAP, 3.5% for CO, and 3.95% for SVO(2)). The computer model was found to accurately predict the experimental results observed using LBNP. The model should be explored as a platform for studying concepts and physiologic mechanisms of hemorrhage including its diagnosis and treatment. PMID- 19804938 TI - Low dose nitrite enhances perfusion after fluid resuscitation from hemorrhagic shock. AB - This study determines the systemic and microvascular hemodynamic consequences of administering a low dose sodium nitrite after fluid resuscitation from hemorrhagic shock. Hemodynamic responses to hemorrhagic shock and resuscitation were studied in the hamster window chamber model. Moderated hemorrhage was induced by arterial controlled bleeding of 50% of the blood volume (BV) and the hypovolemic state was maintained for 1h. Volume restitution was performed by infusion of 25% of BV using Hextend (6% Hetastarch 670kDa in lactated electrolyte solution) 10min after fluid resuscitation 100microl of specific concentrations of sodium nitrite were infused. The experimental groups were named based on the nitrite concentration used, namely: 0microM, 10microM and 50microM. Systemic parameters, microvascular hemodynamics and capillary perfusion (functional capillary density, FCD) were followed during entire protocol. Exogenous 10microM nitrite maintained systemic and microhemodynamic conditions post fluid resuscitation from hemorrhagic shock, compared to 50microM or no nitrite. A moderated increase in plasma nitrite during the early phase of resuscitation reversed arteriolar vasoconstriction and increased capillary perfusion and venous return, improving central cardiac function. Nitrite effects on resistance vessels, directly influenced intravascular pressure redistribution, sustained blood flow, and prevented tissue ischemia. In conclusion, increasing nitrite plasma bioavailability after fluid resuscitation from hemorrhagic shock can be a potential therapy to enhance microvascular perfusion and to improve overall outcome. PMID- 19804939 TI - Characteristics and outcome of prehospital paediatric tracheal intubation attended by anaesthesia-trained emergency physicians. AB - AIM: To collect data regarding prehospital paediatric tracheal intubation by emergency physicians skilled in advanced airway management. METHODS: A prospective 8-year observational study of a single emergency physician-staffed emergency medical service. Self-reporting by emergency physicians of all children aged 0-14 years who had prehospital tracheal intubation and were attended by either anaesthesia-trained emergency physicians (group 1) or by a mixture of anaesthesia and non-anaesthesia-trained emergency physicians (group 2). RESULTS: Eighty-two out of 2040 children (4.0%) had prehospital tracheal intubation (58 in group 1). The most common diagnoses were trauma (50%; in school children, 73.0%), convulsions (13.4%) and SIDS (12.2%; in infants, 58.8%). The overall tracheal intubation success rate was 57 out of 58 attempts (98.3%). Compared to older children, infants had a higher number of Cormack-Lehane scores of 3 or 4, "difficult to intubate" status (both 3 out of 13; 23.1%) and a lower first attempt success rate for tracheal intubation (p=0.04). Among all 82 children 71 (86.6%) survived to hospital admission and 63 (76.8%) to discharge. Of the 63 survivors, 54 (85.7%) demonstrated a favourable or unchanged neurological outcome (PCPC 1-3). The survival and neurological outcomes of infants were inferior compared to older children (p<0.001). On average an emergency physician performed one prehospital tracheal intubation in 3 years in a child and one in 13 years in an infant. CONCLUSIONS: Anaesthesia-trained emergency physicians working in our system report high success rates for prehospital tracheal intubation in children. Survival and neurological outcomes were considerably better than reported in previous studies. PMID- 19804940 TI - The use of D-dimer with new cutoff can be useful in diagnosis of venous thromboembolism in pregnancy. AB - OBJECTIVE: D-dimer testing has an important role in the exclusion of acute venous thromboembolism (VTE) in the nonpregnant population. Establishing D-dimers role in the diagnosis of VTE in pregnancy is hampered because of the substantial increase of D-dimer throughout gestational age. STUDY DESIGN: In a prospective study we followed 89 healthy pregnant women to establish the reference range of D dimer for each trimester. D-dimer testing was also performed in 12 women with clinical suspicion of VTE and their results were compared with the established new reference range of D-dimer, and with the recorded ultrasound findings. RESULTS: In the first trimester, 84% women from reference group had normal D dimer, in the second 33%, and by the third trimester only 1%, which suggests that D-dimer has no practical diagnostic use in ruling out VTE if the threshold of 230 ng/mL for abnormal is used. All pregnant women with thrombosis who had positive ultrasound findings also had statistically significant elevation of the D-dimer level, considering the established reference range of the corresponding trimester. We found 100% sensitivity of D-dimer test. A women developed thrombosis in the first trimester had 6.7-7.6 time higher level of D-dimer than the mean value in the reference group, and in the third trimester thrombotic women had 2.0-3.8 time higher level of D-dimer, p<0.0001. CONCLUSION: D-dimer test with the new threshold for: the first of 286, the second of 457 and the third trimester of 644 ng/mL can be useful in diagnosis of pregnancy related VTE. PMID- 19804941 TI - Human melanoma cells expressing the alphavbeta3 integrin are partially protected from necrotic cell death induced by dynamic matrix detachment. AB - Anchorage-independence is a hallmark of metastatic cancer cells. In previous studies we characterized a novel model for anchorage-independence employing dynamic matrix detachment (DMD) using rotation in low shear stress conditions. We observed that in contrast to the classical apoptosis-inducing static matrix detachment (SMD) model, the venous circulation-mimicking DMD model induced necrosis in transformed cells. In the current study we revisited the mechanism of DMD-induced cell death and evaluated the contribution of alphavbeta3 integrin overexpression in human melanoma cells to anchorage-independence in DMD. DMD cell culture induced primarily necrosis in the melanoma cells studied. alphavbeta3, but not the control related alphaIIbbeta3 integrin, could confer survival advantage in DMD. While apoptosis was unaffected, constitutive, unligated alphavbeta3 overexpression was associated with attenuation of necrosis in DMD. alphavbeta3 overexpressing melanoma cells manifested AKT activation that was independent of DMD conditions. Furthermore, while a small molecular inhibitor of AKT phosphorylation induced apoptosis in adherent cells, in DMD conditions it had no effect on cell outcome. Thus, alphavbeta3-overexpressing melanoma cells are partially protected from DMD-induced cell death in an apoptosis-independent mechanism. This finding may be one of the factors accounting for anchorage independence in circulating metastatic melanoma cells. PMID- 19804942 TI - Detection and identification of Tetratrichomonas in a preputial wash from a bull by PCR and SSCP. AB - Motile Tritrichomonas foetus-like trichomonads were found during microscopic examination of a wet mount sample of a preputial wash collected from a bull. Staining of the organisms with a modified Wright-Giemsa stain revealed that several had four anterior flagella of unequal length instead of the three anterior flagella of equal length characteristic of T. foetus. Limited propagation of these organisms was achieved in InPouch medium but no growth occurred in modified Diamond's medium. The 5.8S rRNA gene and the flanking internal transcribed spacers were amplified from the trichomonad gDNA of two preputial wash samples and a fecal sample taken from the affected bull. Amplicons were subjected to single-strand conformation polymorphism (SSCP) analyses. The SSCP banding patterns of the amplicons from gDNA of the preputial wash samples were different from those of a T. foetus control sample. These unknown trichomonads were not detected in the fecal sample. The gDNA extracted from preputial washes was also subjected to PCR using primers developed to amplify the 16S rDNA of the non-T. foetus trichomonads, Tetratrichomonas and Pentatrichomonas spp. Amplicons were produced from the gDNA of the two preputial washes but not from the T. foetus gDNA control sample. The 16S rDNA sequences obtained from the trichomonads in the two preputial washes samples were 100% similar to that of a Tetratrichomonas species previously isolated from an Angus bull from the United States. PMID- 19804943 TI - Aging males' symptoms in relation to the genetically determined androgen receptor CAG polymorphism, sex hormone levels and sample membership. AB - Late-onset hypogonadism describes the co-occurrence of a range of physical, psychological and sexual symptoms in aging men, with the implication that these symptoms are caused by androgen deficiency. Previous investigations examined mostly population samples and did not take into account the testosterone modulating effects of the genetically determined CAG repeat polymorphism (CAGn) of the androgen receptor (AR) gene. This is the first study which investigates aging male symptoms (AMS) in relation to the genetically determined androgen receptor CAG polymorphism, estradiol and testosterone levels in men > or =50 years of age in a healthy population sample (n=100), outpatients of an andrological department (n=76) who presented with sexual and "aging male" symptoms and a psychosomatic/psychiatric sample (n=120) who presented with various psychological and medically unexplained somatic complaints. Although the population sample was significantly older than the two patient groups, they reported significantly fewer AMS and had higher testosterone levels and shorter CAG repeats of the AR. Regression analysis revealed influences of CAGn on the AMS global score and the psychological and somatic subscale only in the two patient samples, while testosterone had some impact on the sexual subscale. Our results suggest that the so-called aging male symptoms show a certain association to androgenicity, but that they are rather unspecific and of multifactorial origin. Other factors contributing to AMS need further clarification. PMID- 19804944 TI - Watching a previous victory produces an increase in testosterone among elite hockey players. AB - Previous research indicates that testosterone concentrations are highly responsive to human competitive interactions and that winners have elevated testosterone concentrations relative to losers. Also, there is some evidence that simply observing others compete can have a similar effect on the endocrine system. Here, in two studies, we examined the extent to which elite male hockey players would demonstrate an increase in testosterone concentrations after watching themselves engaged in a previous successful competitive interaction. Results indicated that watching a previous victory produced a significant increase in testosterone concentrations (42-44% increase), whereas watching a previous defeat or a neutral video did not produce a significant change in testosterone (17% and 6%, respectively). Given that natural fluctuations in testosterone have been shown to influence future competitive and aggressive behaviours, the current studies may have important practical implications for individuals involved in competitive sports. PMID- 19804946 TI - Dental caries: cola, calor or calories? PMID- 19804945 TI - Correlates of smoking cessation self-efficacy in a community sample of smokers. AB - While numerous studies show that higher levels of smoking cessation self-efficacy predicts motivation to quit smoking and successful smoking cessation, few studies have evaluated factors related to smoking cessation self-efficacy that could be targets of behavioral interventions to promote greater confidence to quit smoking. This study, using a large community sample of smokers enrolled in a smoking cessation treatment program, evaluated potential associations between self-efficacy to quit smoking and demographic (e.g., age, race), smoking-related (e.g., rate, cessation history, past use of treatments), and psychosocial (e.g., stress, cue reactivity, self-medication smoking) variables. The results indicated that Hispanic-American smokers, relative to smokers of other racial/ethnic groups, report significantly lower self-efficacy to quit smoking when facing internal stimuli (e.g., feeling depressed), as do smokers who report that they have little confidence to control abstinence-induced symptoms (F(9,576)=6.9, p<.001). The results also indicated that smokers who reported that they have little confidence to control abstinence-induced symptoms and report high smoking urge reactivity to situations that illicit positive affect (e.g., at a bar, with coffee, at a party) report lower self-efficacy to quit smoking when facing external stimuli (e.g., during a celebration; F[7,600]=9.05, p<.05). These findings can be used to refine behavioral smoking cessation interventions to increase self-efficacy to quit smoking. PMID- 19804947 TI - First trimester curtailment of iron absorption: innate suppression of a teratogen? AB - In human pregnancies, maternal absorption of iron is markedly curtailed in the first trimester. In a murine model, iron was teratogenic in the analogous embryonic period. Although iron is a weak mutagen, it is a powerful oxidant and a catalyst of formation of hydroxyl radicals. Studies are needed to determine if there might be an association of first trimester iron supplementation with miscarriage/fetal abnormalities. PMID- 19804948 TI - Focal salvage guided by T2-weighted and dynamic contrast-enhanced magnetic resonance imaging for prostate cancer recurrences. AB - PURPOSE: Salvage treatment of the entire prostate for local recurrent cancer after primary radiotherapy is associated with high toxicity rates. Our goal was to show that, using dynamic contrast-enhanced magnetic resonance imaging (DCE MRI) for the visualization of a recurrence, focal salvage treatment can be performed, with, potentially, a reduction in toxicity. METHODS AND MATERIALS: We performed MRI, including a DCE sequence, in 7 patients with biopsy-proven locally recurrent prostate cancer. The specific regions of interest suspect for containing tumor were delineated using DCE and T(2)-weighted MRI scans. Subsequently, focal salvage high-dose-rate brachytherapy plans were created to illustrate the principle of focal salvage. Total salvage treatment plans were also created for comparison. RESULTS: The transfer constant (K(trans)) values from the DCE were 0.33-0.67 min(-1) for areas suspect for tumor and 0.07-0.25 min(-1) for normal tissue. In 4 cases, a focal salvage plan could be generated; 93-100% of the gross tumor volume was covered with the prescribed dose, with relative sparing of the bladder, rectum, and urethra. In the total salvage plans, 24-53% of the gross tumor volume was covered, and the organs at risk received high doses. In 3 cases, a focal salvage plan could not be created because of multifocal tumor, seminal vesicle extension, or capsular extension. CONCLUSION: Focal salvage treatment plans can be created in patients with local recurrent prostate cancer after radiotherapy. DCE-MRI supports the localization of the target area. This could lead to less toxicity in patients with local recurrent prostate cancer. PMID- 19804949 TI - Embryonic resorption, litter size and sex ratio in the grasscutter, Thryonomys swinderianus. AB - This study was conducted using 210 whole uteri collected from hunters' kills from July 2006 to March 2007 at Esiam and Sokode in the Ekumfi and Ho Districts respectively; and data on 148 dams from eight farms in the Greater Accra Region of Ghana from January to December 2007. The objective of the study was to investigate the effects of embryonic resorption, litter size and parity on sex ratio in the grasscutter. Embryonic resorption rate, which occurred at all stages of pregnancy, ranged between 41.3% and 47.2%, and was not biased towards male or female offspring. The rate of resorption also had no effect on sex ratio (r= 0.011, P=0.396). The study also showed that litter size in the grasscutter could be improved if embryo resorption is controlled. Primiparous dams had offspring with a sex ratio of 0.43+/-0.020 which was not significantly different from dams of parities 3 (0.47+/-0.040) and > or =4 (0.53+/-0.053). Parity 2 dams, however, had a significantly higher proportion of males (0.54+/-0.024). There was a tendency towards a decreasing proportion of males as litter size increased but the overall sex ratio observed in the farmed grasscutter (0.48) did not deviate significantly from the theoretical sex ratio of 0.50. PMID- 19804950 TI - CT characteristics of resolving ground-glass opacities in a lung cancer screening programme. AB - PURPOSE: This study aimed at evaluating the computed tomography (CT) characteristics of resolving localized ground-glass opacities (GGOs) in a screening programme for lung cancer. MATERIAL AND METHODS: 280 patients at high risk for lung cancer (221 men, 59 women; mean age, 58.6 years), divided into four groups (lung cancer history (n = 83), head and neck cancer history (n = 63), symptomatic (n = 88) and asymptomatic (n = 46) cigarette smokers), were included in a prospective trial with annual low-dose CT for lung cancer screening. We retrospectively reviewed all localized GGOs, analyzed the CT characteristics on initial CT scans and changes during follow-up (median 29.1 months). Variables associated with resolution of GGOs were tested using chi-square or Mann-Whitney tests. RESULTS: A total of 75 GGOs were detected in 37 patients; 54.7% were present at baseline and 45.3% appeared on annual CT. During follow-up, 56.2% persisted and 43.8% disappeared. The resolving localized GGOs were significantly more often lobular GGOs (p = 0.006), polygonal in shape (p = 0.02), mixed (p = 0.003) and larger (p < 0.0001) than non-resolving localized GGOs. CONCLUSION: Localized GGOs are frequent and many disappeared on follow-up. CT characteristics of resolving GGOs show significant differences compared to persistent ones. This study emphasizes the importance of short-term CT follow-up in subjects with localized GGOs. PMID- 19804951 TI - Discrimination between invasive pulmonary aspergillosis and pulmonary lymphoma using CT. AB - OBJECTIVE: The purpose was to assess the characteristic CT features of invasive pulmonary aspergillosis (IPA) and pulmonary lymphoma (PL) and to analyze the potential to distinguish the two entities using CT. METHODS: The CT images of 70 patients with either proven IPA (n = 35) or PL (n = 35) were evaluated retrospectively and independently by two radiologists (reader 1 [R1] and reader 2 [R2]), analyzing images for presence, number and characteristics of pulmonary nodules and masses, ground-glass opacities, consolidations and other interstitial changes. RESULTS: Interreader agreement was moderate (4/33 CT features), good (9/33) or excellent (20/33). Pulmonary nodules (P = 0.045 [R1], P = 0.001 [R2]), nodules with spiculated outer contours (P < 0.001 [R1], P = 0.001 [R2]), nodules with a halo sign (P < 0.001 [R1 + R2]), nodules with homogeneous (P = 0.030 [R1], P = 0.006 [R2]) and inhomogeneous (P = 0.001 [R1], P < 0.001 [R2]) attenuation patterns, nodules with cavitation (P = 0.006 [R1], P = 0.003 [R2]) and wedge shaped, pleural-based consolidations (P < 0.001 [R1 + R2]) occurred significantly more often in patients with IPA, while masses without a halo sign (P = 0.03 [R1], P = 0.01 [R2]), lobar consolidations with bronchogram (P = 0.02 [R1 + R2]) and consolidations with homogeneous attenuation patterns (P < 0.001 [R1 + R2]) were found significantly more frequent in PL-patients. CONCLUSIONS: Those CT features can therefore be considered suggestive for either IPA or PL. However, in most cases the diagnosis cannot be made based on CT findings solely because no single feature gained a high sensitivity and specificity concomitantly. Furthermore, the logistic regression did not show a combination that was significantly better than the best univariate predictor. PMID- 19804952 TI - Metabolism of sanguinarine in human and in rat: characterization of oxidative metabolites produced by human CYP1A1 and CYP1A2 and rat liver microsomes using liquid chromatography-tandem mass spectrometry. AB - The quaternary benzo[c]phenanthridine alkaloid, sanguinarine (SA), has been detected in the mustard oil contaminated with Argemone mexicana, which produced severe human intoxications during epidemic dropsy in India. Today, SA metabolism in human and in rat has not yet been fully elucidated. The goal of this study is to investigate the oxidative metabolites of SA formed during incubations with rat liver microsomes (RLM) and recombinant human cytochrome P450 (CYP) and to tentatively identify the CYP isoforms involved in SA detoxification. Metabolites were analyzed by liquid chromatography combined with electrospray ionization tandem mass spectrometry. Up to six metabolites were formed by RLM and their modified structure has been proposed using their mass spectra and mass shifts from SA (m/z 332). The main metabolite M2 (m/z 320) resulted from ring-cleavage of SA followed by demethylation, whereas M4 (m/z 348) is oxidized by CYP in the presence of NADPH. The diol-sanguinarine metabolite M6 (m/z 366) formed by RLM might derive from a putative epoxy-sanguinarine metabolite M5 (m/z 348). M4 and M6 could be detected in rat urine as their respective glucuronides. 5,6 Dihydrosanguinarine is the prominent derivative formed from SA in cells expressing no CYP. Oxidative biotransformation of SA was investigated using eight human CYPs: only CYP1A1 and CYP1A2 displayed activity. PMID- 19804953 TI - Validation of handgrip strength and endurance as a measure of physical function and quality of life in healthy subjects and patients. AB - OBJECTIVE: Handgrip strength (HGS) is often used as a bedside measurement of muscle function in the hospital setting. The aim of this study was to investigate the extent to which HGS, endurance, and work (force during endurance x time) are related to physical function as measured by mobility and physical activity (PA) in young, healthy volunteers. Further, the relations between HGS, mobility, PA and quality of life (QoL) in patients were investigated. METHODS: Ninety-two healthy subjects (45% men, mean age 30 y) and 45 patients (56% men, mean age 55 y) were assessed for HGS, handgrip endurance, and handgrip work, mobility (timed up-and-go test), and PA (Baecke questionnaire or Bouchard activity diary). The patients were further assessed for QoL (SF-36). RESULTS: There was a correlation between HGS and mobility in healthy subjects (r=-0.31, P=0.0028) and patients (r= 0.59, P<0.0001). Further, HGS and mobility were related to physical and mental component summary scores of QoL in patients. There was also a relation between HGS and PA in healthy female subjects and male patients. CONCLUSION: Handgrip strength is a valid measurement of mobility and QoL in patients and of PA in healthy female subjects and male patients. Handgrip endurance and work were not found to be valid measurements of mobility and PA in healthy subjects or of QoL in patients. PMID- 19804954 TI - No difference in the 24-hour interstitial fluid glucose profile with modulations to the glycemic index of the diet. AB - OBJECTIVE: Reducing the glycemic index (GI) of the diet may decrease metabolic risk, primarily through reduction and stabilization of blood glucose. The objective of this research was to investigate whether incorporation of lower or higher GI foods into mixed meals had different effects on daylong glucose profiles, measured in interstitial fluid by a continuous glucose-monitoring system. METHODS: The study was a randomized, balanced, two-way crossover intervention of 2 x 1-wk periods of lower and higher GI diets. Participants were 12 overweight healthy adult women (mean body mass index +/- standard deviation 27.5+/-2.3 kg/m(2)). Changes in GI were achieved through substitution of key staple carbohydrate-rich foods. After a 4-d run-in on each dietary regimen, participants wore the continuous glucose-monitoring system over 2 d of identical controlled feeding in the laboratory, separated by 1 d of ad libitum consumption at home. RESULTS: On controlled days, diets differed in GI by 15 U and provided equal energy, macronutrients, and fiber. On ad libitum days, diet diaries revealed a difference in GI of 14+/-1 U (mean +/- standard error), with no detectable difference in energy, macronutrient, or fiber intake. No differences were observed in glucose profiles between higher and lower GI interventions in the controlled or ad libitum setting. There was significant agreement in area under the glucose curve on repeated controlled feeding days (intraclass correlation 0.75). CONCLUSION: This study indicates that a difference in dietary GI of 14-15 U is insufficient to alter daylong glycemia as measured in interstitial fluid by the continuous glucose-monitoring system. PMID- 19804955 TI - Food security measurement in cultural pluralism: missing the point or conceptual misunderstanding? AB - OBJECTIVE: Food Security has become a global concern, yet its measurement has varied considerably across disciplines and countries. We examined the current discrepancies in the definitions of food security and propose a framework for understanding and measuring food security. METHODS: This conceptual review draws from a range of works published in Medline and the gray literature to advance the understanding of food security concepts. We begin by examining the historical background of food security and then move on to examine its various definitions and interpret food through cultural lenses in terms of food access and utilization. We finish by examining various measurements and indicators of food security and reviewing implications for public health. RESULTS: We argue that the reliance on coping strategies as surrogate measurements of food insecurity without taking into account the social, cultural, and political contexts in which they occur is misleading, and viewing food insecurity solely from a food access or availability perspective, without taking into account food utilization and asset creation as pillars of food security, paints an incomplete picture. Although this review does not claim to provide solutions to the discrepancies in the conceptual definition of food security, it attempts to highlights areas of concern and provide a way forward. CONCLUSION: When coping strategies are used as an indicator of food insecurity, they need to be culturally relevant and focus tested, and together with objective measurements of nutritional outcomes, would allow policy makers to make evidence-based decisions to inform social and nutrition policies. PMID- 19804956 TI - Role of an electronic armband in motor function monitoring in patients with Parkinson's disease. AB - OBJECTIVE: Levodopa replacement still is the gold standard for the management of Parkinson's disease (PD). Long-term treatment with levodopa is frequently associated with motor fluctuations. A low-protein (LP) dietary regimen has proved to be effective in reducing this adverse effect, but has been associated with weight loss, probably due to increased energy expenditure. A new wearable device (SenseWear Armband [SWA]) has recently been introduced into clinical practice. It is designed to monitor physical activity continuously and provide estimates of energy consumption. We assessed its role in measuring the effects of dietary regimens on motor function in PD. METHODS: Six patients with levodopa-treated PD and motor fluctuations were asked to follow a balanced diet (protein 1g x kg(-1) x d(-1)) for 7 d and then to cross over to a isocaloric LP (protein 0.7 g x kg( 1) x d(-1)) dietary regimen. Total daily energy expenditures, physical activity, number of steps, and metabolic rate were assessed continuously (14 d) by the SWA. Motor control was evaluated by daily diaries. RESULTS: The SWA proved that, during the LP diet, mean total daily energy expenditure was higher (P<0.05) and so were physical activity (P=0.05) and average metabolic rate (P=0.01), despite no change in the number of steps. The duration of periods with dyskinesias was also increased (P<0.05). These data support the role of upper-extremity involuntary movements in increasing total daily energy expenditure during an LP diet. CONCLUSION: The SWA may help in monitoring patients with PD because it can assist in evaluating motor response to treatment and changes in physical activity and daily calorie needs. PMID- 19804957 TI - Atraumatic placement of circummandibular wires: a technical note. AB - Fractures of the mandible are relatively uncommon in children as compared to that of adults. When they occur, the problems associated with their management are complicated due to the presence of the tooth bud and potential growth disturbance. Various management protocols are discussed in the literature. Most authors agree that the ideal method is to use an acrylic splint because it is easy to place and reduces the risk to jaw growth. These splints are secured to the reduced jaw by circummandibular wiring, which is a relatively simple technique. The authors describe their experience with a new atraumatic technique for the placement of circummandibular wires using a 16 gauge intravenous cannula stillete. PMID- 19804958 TI - Posterior dislocation in intact mandibular condyle: an unusual case. AB - Dislocation of the temporomandibular joint (TMJ) represents 3% of all reported dislocated joints. Traditionally, dislocation of the TMJ occurs when the mandibular condyle is displaced anteriorly beyond the articular eminence. Traumatic dislocation of the condyle into the middle cranial fossa is well defined in the literature, but posterior dislocation without fracture is rarely described. This report documents a case of young male with posterior dislocation of the intact mandibular condyle after facial trauma. The clinical symptoms, diagnosis and treatment are discussed, and a critical review of the literature is provided. PMID- 19804959 TI - Contributions of peripheral inflammation to seizure susceptibility: cytokines and brain excitability. AB - Inflammation is an important factor in the pathophysiology of seizure generation and epileptogenesis. While the role of CNS inflammation is well acknowledged as an important factor in seizure pathophysiology, less is known about the role of peripheral inflammation. Systemic inflammation induces a mirror inflammatory response in the brain that might have transient or long-term effects on seizure susceptibility. The focus of our laboratory research is the study of the interaction of systemic inflammatory events with neuronal excitability and seizure susceptibility. In this paper we provide a review of our findings and discuss possible mechanisms. PMID- 19804961 TI - Rectal filling at planning does not predict stability of the prostate gland during a course of radical radiotherapy if patients with large rectal filling are re-imaged. AB - AIMS: It has been suggested that large rectal filling is associated with an increased risk of prostate motion in radiotherapy. The aim of the present study was to determine if there is a correlation between rectal distension on planning computed tomography and the intrafraction and interfraction stability of the prostate gland during a course of radical radiotherapy for prostate cancer if a protocol was used to rescan patients with excessive rectal diameter during planning. MATERIALS AND METHODS: The computed tomography planning scans of 89 patients with adenocarcinoma of the prostate treated with conformal radiotherapy were reviewed. All patients had three gold seed fiducial markers implanted into the prostate before planning computed tomography. About one in five patients had repeat computed tomography because their rectum was judged to be too large at the time of the first planning computed tomography. Rectal distension was assessed on planning computed tomography using outlines following European Organization for Research and Treatment of Cancer guidelines by measuring the rectal volume, the average cross-sectional area and the mean anterior-posterior diameter of the rectum. Daily kV images were obtained before and after treatment delivery to determine positional matching of the fiducial markers in the superior-inferior, anterior-posterior and right-left dimensions. RESULTS: In total, 2860 pre- and post-treatment daily kV image pairs were obtained of 89 patients (average 32.1 image pairs per patient). The median rectal cross-sectional area was 7.3cm(2) (range 2.8-17.1), the median rectal volume was 54.8cm(3) (range 20.9-128.2), and the median anterior-posterior rectal diameter was 3.03cm (range 1.58-8.30). Unifactor linear regression models showed no statistically significant relationship between intra- and interfraction prostate stability and rectal volume on planning computed tomography. CONCLUSIONS: No statistically significant relationship between rectal distension on planning computed tomography and the intra- and interfraction stability of the prostate gland was identified if patients with a large rectal volume were rescanned for planning. PMID- 19804960 TI - NPY mRNA expression in the prefrontal cortex: Selective reduction in the superficial white matter of subjects with schizoaffective disorder. AB - BACKGROUND: Alterations in the inhibitory circuitry of the dorsolateral prefrontal cortex (DLPFC) in schizophrenia include reduced expression of the messenger RNA (mRNA) for somatostatin (SST), a neuropeptide present in a subpopulation of gamma-aminobutyric acid (GABA) neurons. Neuropeptide Y (NPY) is expressed in a subset of SST-containing interneurons and lower levels of NPY mRNA have also been reported in schizophrenia spectrum disorders. However, whether the alterations in these two transcripts identify the same, particularly vulnerable, subset of GABA neurons has not been examined. METHODS: We used in situ hybridization to quantify NPY mRNA levels in DLPFC gray and white matter from 23 pairs of subjects with schizophrenia or schizoaffective disorder and matched normal control subjects; results were compared to those from a previous study of SST mRNA expression in the same subjects. RESULTS: In contrast to SST mRNA, NPY mRNA levels were not significantly lower in the gray matter of subjects with schizophrenia or schizoaffective disorder. However, NPY, but not SST, mRNA expression was significantly lower in the superficial white matter of subjects with schizoaffective disorder. CONCLUSION: These findings suggest that the alterations in SST-containing interneurons in schizophrenia and schizoaffective disorder are selective for the subset that do not express NPY mRNA, and that lower NPY mRNA expression in the superficial white matter may distinguish subjects with schizoaffective disorder from those with schizophrenia. PMID- 19804962 TI - Ultrastructural and DNA damaging effects of lead nitrate in the liver. AB - A ubiquitous environmental toxicant - lead is known to affect several organ systems. This study was designed to investigate the effects of lead nitrate exposure on liver structure and DNA fragmentation. Adult male Wistar rats were treated orally with lead nitrate at the dose levels of 0%, 0.5% and 1% for 60 days and sacrificed on the next day. The liver was processed for thick sections and evaluated after toludine blue staining and by electron microscopy after staining with uranyl acetate and lead citrate. The DNA damage was assessed by DNA fragmentation assay. The liver weight was not significantly affected in the experimental groups. Hepatocyte nuclei were not shrunk, instead lead was mitogenic to hepatocytes as indicated by an increase in the number of binucleated hepatocytes (P<0.05). The number of mitochondria per hepatocyte decreased in a dose-dependent manner (P<0.05). Qualitatively, the necrotic changes such as small to large-sized cytoplasmic vacuoles often displacing the organelles, decrease in hepatocyte microvilli, degeneration of mitochondria, and vacuolar encroachment of nuclei and dilatation of sinusoids were observed. The qualitative changes were induced in a dose-dependent manner. Kupffer cells or Ito cells did not present any notable structural changes. Although the electrophoretic flow of DNA fragments was observed in lead-treated groups, these changes were not significantly different from that in control as evaluated by optical density. In conclusion, lead induces necrotic changes with simultaneous mitogenic activity; however, it does not induce significant DNA damage in the liver. PMID- 19804963 TI - Bilevel ventilation during exercise in acute on chronic respiratory failure: a preliminary study. AB - To determine the immediate effects of bilevel non-invasive ventilation plus oxygen (NIV+O(2)) during exercise compared to exercise with O(2) alone in people recovering from acute on chronic hypercapnic respiratory failure (HRF), a randomised crossover study with repeated measures was performed. Eighteen participants performed six minute walk tests (6MWT) and 16 participants performed unsupported arm exercise (UAE) tests with NIV+O(2) and with O(2) alone in random order. Distance walked increased by a mean of 43.4m (95% CI 14.1 to 72.8, p=0.006) with NIV+O(2) compared to exercise with O(2) alone. In addition, isotime oxygen saturation increased by a mean of 5% (95% CI 2-7, p=0.001) and isotime dyspnoea was reduced [median 2 (interquartile range (IQR) 1-4) versus 4 (3-5), p=0.028] with NIV+O(2). A statistically significant increase was also observed in UAE endurance time with NIV+O(2) [median 201s (IQR 93-414) versus 157 (90-342), p=0.033], and isotime perceived exertion (arm muscle fatigue) was reduced by a mean of 1.0 on the Borg scale (95% CI -1.9 to -0.1, p=0.037) compared with O(2) alone. Non-invasive ventilation plus O(2) during walking resulted in an immediate improvement in distance walked and oxygen saturation, and a reduction in dyspnoea compared to exercise with O(2) alone in people recovering from acute on chronic HRF. The reduction of dyspnoea during walking and arm muscle fatigue during UAE observed with NIV+O(2) may allow patients to better tolerate exercise early in the recovery period. PMID- 19804964 TI - Management of aggressive fibromatosis: can we unravel the maze of treatment options? PMID- 19804965 TI - The mystery of male dominance in oesophageal cancer and the potential protective role of oestrogen. AB - Oesophageal cancer is the sixth most common form of cancer death globally with almost 400,000 deaths annually. More than 90% of all cases are either adenocarcinomas (OAC) or squamous-cell carcinomas (OSCC). There is a strong male predominance with up to 8 and 3 men for every woman affected with OAC and OSCC, respectively. It has been hypothesised that sex hormonal factors may play a role in the development of oesophageal cancer or more specifically that oestrogen prevents such development. This article reviews the available literature on this topic. Basic science studies suggest an inhibitory effect of oestrogen in the growth of oesophageal cancer cells, and a possible mechanism of any oestrogen protection might be mediated through oestrogen receptors. But from the few epidemiological studies in which the hypothesis of oestrogen protection has been tested, no firm conclusions can yet be drawn of the role of oestrogen in human oesophageal cancer aetiology. More evidence from valid and large human studies is needed before any conclusions can be drawn. PMID- 19804967 TI - The role of (18)F-fluorodeoxyglucose positron emission tomography in the diagnosis, staging, and follow-up of cholangiocarcinoma. AB - (18)F-Fluorodeoxyglucose positron emission tomography ((18)FDG-PET) is a new diagnostic technique for the diagnosis and staging of cholangiocarcinoma. For diagnosis of a primary cholangiocarcinoma, (18)FDG-PET seems to be helpful to discriminate between malignant and benign lesions. However, the accuracy of (18)FDG-PET seems to be dependent on the anatomic location, growth pattern, and pathologic characteristics of the lesion. It has been proved that the accuracy of (18)FDG-PET is limited to detection of extrahepatic, infiltrating, and mucinous cholangiocarcinomas. Due to its lower sensitivity, (18)FDG-PET provides complementary rather than confirmative information in the diagnosis of regional lymph node metastasis. In contrast, it has high accuracy in detecting unsuspected distant metastases. The role of (18)FDG-PET in detecting cancer recurrence, monitoring treatment response, and predicting prognosis is still controversial. PMID- 19804966 TI - Prostate cancer mortality in screen and clinically detected prostate cancer: estimating the screening benefit. AB - BACKGROUND: To estimate the benefits of prostate-specific antigen (PSA) screening on prostate cancer (Pca) metastasis and Pca-specific mortality, we compared two populations with a well-defined difference in intensity of screening. METHODS: Between 1997 and 1999, a total of 11,970 men, aged 55-74 years, were included in the intervention arm of the European Randomised Study of Screening for Prostate Cancer (ERSPC) section Rotterdam. Control population consisted of 133,287 men, aged 55-74 years, between 1998 and 1999 in Northern Ireland (NI). Men were followed for Pca incidence, Pca metastasis and cause of death until 31st December 2006. RESULTS: Median age in both groups was 63 years at study entry (p=0.184). In Rotterdam 94.2% of men and in NI 6% of men underwent PSA testing. In Rotterdam, 1153 men (9.6%) were diagnosed with Pca with median baseline PSA of 5.1 ng/ml. In NI, 3962 men (3.0%, p<0.001) were diagnosed with Pca with median baseline PSA of 18.0 ng/ml (p<0.001). The relative risk of Pca metastasis during observation in the intervention population compared to control population was 0.47 (95% confidence interval (CI), 0.35-0.63; p<0.001). The relative risk of Pca specific mortality was also lower in the intervention population compared to the control population after a median follow-up of 8.5 years: 0.63 (95% CI, 0.45 0.88; p=0.008); absolute mortality reduction was 1.8 deaths per 1000 men. CONCLUSIONS: A relative reduction in Pca metastasis of 53% and Pca mortality of 37% was observed in the intervention population after 8.5 years of observation. The impact of overdiagnosis, quality of life benefits and cost-effectiveness need to be assessed before population-based PSA screening can be recommended. PMID- 19804968 TI - Methane production from rice straw pretreated by a mixture of acetic-propionic acid. AB - Rice straw was treated with a mixed solution of acetic acid and propionic acid to enhance its biodegradability. The effect of acid concentration, pretreatment time, and the ratio of solid to liquid on the delignification performance of rice straw were investigated. It was found that the optimal conditions for hydrolysis were 0.75 mol/L acid concentration, 2h pretreatment time and 1:20 solid to liquid ratio. Batch methane fermentation of untreated rice straw, pretreated rice straw, and the hydrolysates (the liquid fraction) of pretreatment were conducted at 35 degrees C for 30 days, and the results indicated that methane production of rice straw can be enhanced by dilute organic acid pretreatment. Moreover, most of the acid in hydrolysates can also be converted into methane gas. PMID- 19804969 TI - The discovery and optimisation of benzazepine sulfonamide and sulfones as potent agonists of the motilin receptor. AB - Optimisation of a series of benzazepine sulfonamide hit compounds identified from high throughput screening led to the discovery of a new series of tractable, potent motilin receptor agonists. PMID- 19804970 TI - Design of novel alpha7-subtype-preferring nicotinic acetylcholine receptor agonists: application of docking and MM-PBSA computational approaches, synthetic and pharmacological studies. AB - In the search for nicotinic acetylcholine receptor (nAChRs) agonists with a selective affinity for the homomeric alpha7 channels, we carried out the virtual screening of a test set of potential nicotinic ligands, and adopted a simplified MM-PBSA approach to estimate their relative binding free energy values. By means of this procedure, previously validated by a training set of compounds, we reached a realistic compromise between computational accuracy and calculation rate, and singled out a small group of novel structurally related derivatives characterized by a promising theoretical affinity for the alpha7 subtype. Among them, five new compounds were synthesized and assayed in binding experiments at neuronal alpha7 as well as alpha4beta2 nAChRs. PMID- 19804971 TI - Discovery and optimization of CRTH2 and DP dual antagonists. AB - A series of phenylacetic acid derivatives was discovered as CRTH2 antagonists. Modification of the series led to compounds that are also antagonists of DP. Since activation of CRTH2 and DP are believed to play key roles in mediating responses of asthma and other immune diseases, this series was optimized to increase the dual antagonistic activities and improve pharmacokinetic properties. These efforts led to selection of AMG 009 as a clinical candidate. PMID- 19804972 TI - Solid-phase synthesis of a library based on biphenyl-containing trypsin-like serine protease inhibitors. AB - The solid-phase synthesis of a library based on an unusual biphenyl-containing trypsin-like serine protease inhibitor is described. Key to this effort was the synthesis of a highly functionalized aryl boronic acid reagent which required the development of a novel and efficient method to convert a triflate to a pinacolboronate in large scale. PMID- 19804973 TI - Antioxidant reactivity toward nitroxide probes anchored into human serum albumin. A new model for studying antioxidant repairing capacity of protein radicals. AB - A new strategy to evaluate accessibility of antioxidants to radical proteins has been developed using nitroxide prefluorescent probes anchored into human serum albumin (HSA). Binding association constants for the nitroxide probes C(343)T and QT with HSA were 5 x 10(4) and 9 x 10(4)M(-1), respectively. Rate constants for the nitroxide reduction by antioxidants in HSA were determined finding k(HSA)/k(buffer) ratio of 0.8, 1.9, and 0.075 for ascorbic acid, Trolox, and caffeic acid, respectively, for the nitroxide C(343)T reduction. PMID- 19804974 TI - Systematic study on the broad nucleotide triphosphate specificity of the pyrophosphorylase domain of the N-acetylglucosamine-1-phosphate uridyltransferase from Escherichia coli K12. AB - N-Acetylglucosamine-1-phosphate uridyltransferase (GlmU) from Escherichia coli K12 is a bifunctional enzyme that catalyzes both the acetyltransfer and uridyltransfer reactions in the prokaryotic UDP-GlcNAc biosynthetic pathway. In this study, we report the broad substrate specificity of the pyrophosphorylase domain of GlmU during its uridyltransfer reaction and the substrate priority is ranked in the following order: UTP > dUTP > dTTP >> CTP > dATP/dm(6) ATP. This pyrophosphorylase domain of GlmU is also a tool to synthesize UDP-GlcNAc analogs, two examples of which were synthesized herein in multiple mg scale in vitro. PMID- 19804975 TI - Introduction of intravenous formulations of the antiepileptic drugs levetiracetam and valproate to the Australian market. PMID- 19804976 TI - In vitro stability of open wedge high tibial osteotomy with synthetic bone graft. AB - It has been predicted that significant stress will be applied to the plate and lateral cortical hinge of an osteotomy site when early full weight bearing is commenced after an open wedge high tibial osteotomy. We hypothesized that the stress concentration on the plate or at the lateral cortical hinge would be reduced by inserting bone substitutes into the osteotomy gap. Two different types of tibia model were investigated: Group A, fixation with TomoFix with the osteotomy site left as an open space; and Group B, two beta-TCP wedges are inserted into osteotomy site and fixed with TomoFix. Stress at five points was measured using strain gauges. Specimens were mounted onto a testing machine with an FTA (femoro-tibial angle) of 170 degrees . Cyclic load tests and an ultimate load test were then performed. The mean stress on the plate was measured at 15.5+/-1.8Mpa in Group A. On the other hand, this value in Group B was only 9.52+/-2.1Mpa and this was a significant difference (P<0.01). The mean stress on the lateral hinge in Groups A and B was 3.31+/-0.5 and 2.49+/-0.2, respectively which was also a significant difference (P<0.05). The mean maximum breaking load in Group A was 2500+/-280N and in Group B 4270+/-420N which was a significant difference (P<0.01). Hence, for OWHTO procedures, the use of beta-TCP wedges and TomoFix is thus likely to improve the initial axial and possibly rotational stability at the osteotomy site in comparison with methods that leave the osteotomy gap open. PMID- 19804977 TI - New approach for the synthesis of [18F]fluoroethyltyrosine for cancer imaging: simple, fast, and high yielding automated synthesis. AB - O-(2-[(18)F]fluoroethyl)-L-tyrosine ([(18)F]FET) is one of the first (18)F labeled amino acids for imaging amino acid metabolism in tumors. This tracer overcomes the disadvantages of [(18)F]fluorodeoxyglucose, [(18)F]FDG, and [(11)C]methionine, [(11)C]MET. Nevertheless, the various synthetic methods providing (18)F[FET] exhibit a big disadvantage concerning the necessity of two purification steps during the synthesis including HPLC purification, which causes difficulties in the automation, moderate yields, and long synthesis times >60 min. A new approach for the synthesis of [(18)F]FET is developed starting from 2 bromoethyl triflate as precursor. After optimization of the synthesis parameters including the distillation step of [(18)F]-FCH(2)CH(2)Br combined with the final purification of [(18)F]FET using a simple solid phase extraction instead of an HPLC run the synthesis [(18)F]FET could be significantly simplified, shortened, and improved. The radiochemical yield (RCY) was about 45% (not decay corrected and calculated relative to [(18)F]F(-) activity that was delivered from the cyclotron). Synthesis time was only 35 min from the end of bombardment (EOB) and the radiochemical purity was >99% at the end of synthesis (EOS). Thus, this simplified synthesis for [(18)F]FET offers a very good option for routine clinical use. PMID- 19804978 TI - Synthesis, molecular docking and biological evaluation of metronidazole derivatives as potent Helicobacter pylori urease inhibitors. AB - Fourteen metronidazole derivatives (compounds 3a-f and 4b-h) have been synthesized by coupling of metronidazole and salicylic acid derivatives. All of them are reported for the first time. Their chemical structures are characterized by (1)H NMR, MS, and elemental analysis. The inhibitory activities against Helicobacter pylori urease have been investigated in vitro and many compounds have showed promising potential inhibitory activities of H. pylori urease. The effect of compounds 4b (IC(50)=26 microM) and 4 g (IC(50)=12 microM) was comparable with that of acetohydroxamic acid, a well known H. pylori urease inhibitor used as a positive control. The experimental values of IC(50) showed that inhibitor was potent urease inhibitor. A docking analysis using the autodock 4.0 program could explain the inhibitory activities of compound 4 g against H. pylori urease. PMID- 19804979 TI - Hybrid pharmacophore design and synthesis of isatin-benzothiazole analogs for their anti-breast cancer activity. AB - A hybrid pharmacophore approach was used to design and synthesize isatin benzothiazole analogs to examine their anti-breast cancer activity. The cytotoxicity of these compounds were determined using three different human breast tumor cell lines, MDA-MB231, MDA-MB468, MCF7, and two non-cancer breast epithelial cell lines, 184B5 and MCF10A. Although all compounds examined were quite effective on all the cancer cell lines examined, the compounds 4-bromo-1 diethylaminomethyl-1H-indole-2,3-dione (2l) and 4-chloro-1-dimethylaminomethyl-3 (6-methyl-benzothiazol-2-ylimino)-1,3-dihydro-indol-2-one (5e) emerged as the most active compounds of this series. Importantly, the cytotoxic effect of 2l was 10-15-fold higher on cancer than non-cancer cells, suggesting that this compound can be very effective for the control of breast cancer with low side effects. Since 2l showed effective cytotoxicity on MCF7 cells and arrested the cells at G2/M at a similar concentration, these two phenomena may be closely correlated. We conclude that the isatin-linked benzothiazole analog can serve as a prototype molecule for further development of a new class of anti-breast cancer agents. PMID- 19804980 TI - Synthesis and biological evaluation of epothilone A dimeric compounds. AB - The preparation and biological evaluation of a novel series of dimeric epothilone A derivatives (1-6) are described. Two types of diacyl spacers were introduced to establish the various dimeric epothilone A constructs. The effect of these compounds on tubulin polymerization and their cytotoxicity against four different cancer cell lines are reported. Several of the newly synthesized compounds inhibit endothelial cell differentiation and endothelial cell migration that are key steps of the angiogenic process. PMID- 19804981 TI - Synthesis, cytotoxicity and human telomerase inhibition activities of a series of 1,2-heteroannelated anthraquinones and anthra[1,2-d]imidazole-6,11-dione homologues. AB - A series of 1,2-heteroannelated anthraquinones and anthra[1,2-d]imidazole-6,11 dione tetracyclic analogues with different side chain were prepared using an various synthetic route via acylation, cyclization, condensation, and intramolecular heterocyclization. Tetracyclic system containing alkyl and aryl, aromatic and heterocyclic, linear and cyclic, polar and apolar, and basic and acids residues were incorporated. They were evaluated for their effects on telomerase activity, hTERT expression, cell proliferations, and in vitro cytotoxicity against NCI's 60 cell line human tumor screen. Compounds 4, 11, 12, 14, 15, 16, 17, 19, 20, 23, 25, and 26 were selected by the NCI for one dose screening program and further studies on 4, 23 and 25 where the curves cross these lines represent the interpolated values to cause 50% growth inhibition (GI(50)), total growth inhibition (TGI) and 50% cell killing (LC(50)), respectively. Further studies did not reveal any compound that showed potent and significant on telomerase inhibitory activity and hTERT repressing ability. Comparative testing of these compounds in the NCI's screen revealed varying levels of potency and differential cytotoxicity, apparently related to the unsaturation levels in and substitution patterns on the core ring system. It appeared that addition of a fourth planar aromatic system to a tricyclic chromophore might enhances potent cytotoxic agents, at a level equivalent to a second side chain in one of the tricyclic series. Although the exact mechanism of how this pharmacophore contributes to its activity is still unclear, however, the group in the extended arm of the tetracyclic system might contribute to proper binding to the residues within the grove of G-quadruplex structure. PMID- 19804982 TI - Synthesis and in vitro evaluation of pteridine analogues as monoamine oxidase B and nitric oxide synthase inhibitors. AB - Monoamine oxidase B (MAO-B) and nitric oxide synthase (NOS) have both been implicated in the pathology of neurodegenerative diseases. In an attempt to design dual-target-directed drugs that inhibit both these enzymes, a series of pteridine-2,4-dione analogues were synthesised. The compounds were found to be relatively weak NOS inhibitors but showed promising MAO-B activity with 6-amino-5 [(E)-3-(3-chloro-phenyl)-prop-2-en-(E)-ylideneamino]-1,3-dimethyl-1H-pyrimidine 2,4-dione and 6-[(E)-2-(3-chloro-phenyl)-vinyl]-1,3-dimethyl-1H-pteridine-2,4 dione inhibiting MAO-B with IC(50) values of 0.602 and 0.314 microM, respectively. The pteridine-2,4-dione analogues thus show promise as scaffolds for the development of potent reversible MAO-B inhibitors and as observed in earlier studies, the most potent inhibitors were obtained with 3-chlorostyryl substitution. PMID- 19804983 TI - Comparison of performance of INAA, RNAA and ion chromatography for the determination of individual lanthanides. AB - Lanthanides represent an interesting group of elements which are steadily gaining importance in science and in industry. Literature data suggest that they may have a role in regulating cellular processes and also in agriculture enhancing plant growth. Only few instrumental methods like NAA, ICP-OES and ICP-MS have potential for the determination of low levels of all lanthanides, but in practice they often have to be combined with suitable separation/preconcentration methods to achieve maximum number of elements being determined and to assure good accuracy and precision. In this work an attempt has been made to compare the performance of both variants of NAA: purely instrumental activation analysis (INAA) and radiochemical mode (RNAA) with pre- and post-irradiation group separation for the determination of individual lanthanides with the special emphasis on rarely determined elements (Gd, Ho, Er, Tm, Lu). This has been done by the analysis of two reference materials with different matrices viz. biological (CTA-OTL-1) and mineral (CTA-FFA-1). The results are compared with those obtained by ion chromatography (IC) employing the same lanthanide group preseparation scheme as in the case of RNAA. PMID- 19804984 TI - Isolation of good quality RNA from a medicinal plant seabuckthorn, rich in secondary metabolites. AB - Medicinal plants are being widely investigated owing to their ability to produce molecules of therapeutic significance. Isolation of good quality RNA is a tedious but primary step towards undertaking molecular biology experiments. However, medicinal plants are rich in secondary metabolites and not amenable to standard RNA isolation protocols involving Guanidine isothiocyanate (GITC). So an RNA isolation protocol from difficult samples (richer in secondary metabolites) is of highest desiderata. Here we propose a new protocol suitable for isolating RNA from plant tissues rich in secondary metabolites. To standard CTAB (Cetyl Trimethyl Ammonium Bromide) buffer, addition of 2% PVPP (polyvinyl polypyrrolidone) and 350 mM beta-mercaptoethanol was found useful. Use of glacial acetic acid (1M) along with ethanol for precipitation after phenolization and chloroform extraction enhanced the RNA yield. This is the first report of using glacial acetic acid in a CTAB based protocol for the precipitation of RNA. This protocol has been validated in medicinal plant Hippophae rhamnoides vern. seabuckthorn, where standard RNA isolation methods involving GITC and TRIZol extraction buffers failed. The RNA isolated by this method was of good quality as gauged by spectrophotometric readings and denaturing agarose gel electrophoresis. To the best of our knowledge, this RNA isolation protocol has never been published before. The RNA thus obtained could be suitably used for the downstream molecular procedures like Reverse Transcription Polymerase Chain Reaction (RT PCR), Real Time-PCR, cDNA library construction, etc. PMID- 19804985 TI - Life course socioeconomic conditions and metabolic syndrome in adults: the Atherosclerosis Risk in Communities (ARIC) Study. AB - PURPOSE: This study examined the effect of childhood, adulthood, and cumulative socioeconomic status (cumSES) on the prevalence of metabolic syndrome (MetS) in middle-aged adults in the Atherosclerosis Risk in Communities Study (1987-1989). METHODS: Participants included 2,461 black and 8,536 white men and women 45 to 64 years of age without diabetes. Socioeconomic status (SES) measures from childhood, early adulthood, and mature adulthood were used to create a cumSES score. Childhood SES, early adult SES, mature adult SES and cumSES scores were grouped into two categories (high/low). Age- and center-adjusted prevalence ratios and 95% confidence intervals for MetS, using the Third Report of the Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (ATP III) criteria, by SES were estimated by log-linear regression for race-sex groups. RESULTS: Black and white women with low childhood SES, early adult SES, mature adult SES or cumSES were more likely to have the MetS than those with high SES. These associations remained after adjustment for physical activity, smoking status, and drinking status. In contrast, there was no association of SES with MetS in men. CONCLUSIONS: Our findings suggest that socioeconomic disadvantage early in life and across the life course influences risk of the MetS in black and white women. PMID- 19804986 TI - Shift work and arteriosclerosis risk in professional bus drivers. AB - PURPOSE: Professional bus drivers are at increased risk for cardiovascular disease, but the underlying causes are unclear. Professional bus drivers often follow shift schedules. Especially, an association between shift work and early manifestations of cardiovascular disease has not been elucidated. Thus we investigated the links between shift work and arteriosclerosis risk in professional bus drivers. METHODS: Questionnaires were administered to 184 bus drivers on demographic characteristics, lifestyle, and occupational history from 5 transportation companies in Taiwan. Brachial-ankle pulse wave velocity (baPWV) was measured using a volume-plethysmographic apparatus. Body mass index, waist circumference, biochemical variables, and blood pressure were also measured. RESULTS: Arteriosclerotic risk factors (age, weekly driving hours, systolic blood pressure, diastolic blood pressure, and insulin level) differed in part among different groups of drivers. Long-term shift drivers had higher baPWV compared to regular drivers and short-term shift drivers (1594 cm/s vs. 1497 and 1432, p<0.01). Our multiple regression model showed that age (p<0.01) and diastolic blood pressure (p<0.01) were positively associated with baPWV in our professional drivers. After adjusting for all covariates, we observed that baPWV increased by 3.6 cm/s for per 1-year increment in years of shift driving. CONCLUSIONS: Long term shift work could increase the risk of arteriosclerosis in professional bus drivers. Larger studies would be necessary to provide further evidence regarding this finding. PMID- 19804987 TI - Social isolation and depression predict 12-month outcomes in the "waiting for a new heart study". AB - BACKGROUND: Identification of modifiable psychosocial characteristics related to survival of heart transplant (HTx) candidates is needed to prevent clinical deterioration and improve prognosis. METHODS: A multi-site, prospective study was conducted with 318 HTx candidates (18% female, 82% male; 53 +/- 11 years of age) newly listed at 17 hospitals in Germany and Austria. Baseline demographic and psychosocial characteristics were assessed by questionnaires. Indicators of disease severity (Heart Failure Survival Score, creatinine, cardiac index) and 12 month outcomes (death, high-urgency HTx, elective HTx, de-listing due to deterioration or improvement) were provided by Eurotransplant. RESULTS: By 12 months, 33 patients died, 83 received an urgent HTx, 30 underwent an elective HTx, and 9 were de-listed due to clinical deterioration and 17 due to improvement. All measures of disease severity predicted outcomes. Controlling for disease severity, the number of social contacts contributed significantly to outcomes, favoring those who improved. Comparing socially isolated patients (<4 social contacts/month) who also had depression scores in the clinical range (high psychosocial risk group; n = 37) to those with >10 social contacts/month without depression (low psychosocial risk group; n = 47) revealed significant differences in the distribution of outcome frequencies (chi-square = 11.2, df = 4, p < 0.04). The high psychosocial risk group was more likely to have died/deteriorated and less likely to have improved than the low psychosocial risk group. CONCLUSIONS: Regardless of disease severity, socially isolated HTx candidates who are also depressed may be at increased risk for clinical deterioration and mortality, indicating a need for psychosocial intervention. PMID- 19804988 TI - Mediastinal radiation and adverse outcomes after heart transplantation. AB - Orthotopic heart transplantation (OHT) may represent the only treatment option for patients with end-stage cardiovascular disease due to mediastinal radiation therapy (MRT). The primary aim of this study was to evaluate the safety and efficacy of OHT in this patient population. We conducted a retrospective, single center cohort study of patients with MRT-associated cardiovascular disease who underwent OHT between January 1987 and September 2008. Nine patients (3 men), aged 46 +/- 11 years at the time of their OHT, were identified. Time from MRT to OHT was 26 +/- 11 years. Lymphoma was the indication for MRT in all patients. Five patients had non-ischemic dilated cardiomyopathy, 2 had ischemic cardiomyopathy and 2 had constrictive pericarditis. Three patients expired in the peri-operative period, whereas another patient died 3 years post-transplant from lung carcinoma. Two additional patients developed a secondary malignancy post transplant. Five patients are still alive at a mean follow-up of 10 +/- 8 years. Early survival rate is poor in patients who undergo OHT for MRT-associated end stage cardiovascular disease. In addition, long-term follow-up shows an elevated incidence of malignancies. Our results raise concern about the safety and efficacy of performing OHT in patients with MRT-associated cardiovascular disease. PMID- 19804989 TI - Interactions among donor characteristics influence post-transplant survival: a multi-institutional analysis. AB - BACKGROUND: Quantification of donor-associated risk in a specific heart transplant recipient is often difficult. Our aim was to identify donor characteristics that affect survival in the contemporary era. METHODS: Between 1990 and 2006, 7,322 patients from 32 centers in the Cardiac Transplant Research Database underwent heart transplantation. Multivariable logistic regression analysis was used to identify donor-associated risk predictors and important interactions between these donor characteristics. Recipient survival was examined using parametric regression analysis in the hazard function domain. RESULTS: Donor characteristics associated with post-transplant death included donor age, donor requirement for vasoactive therapy, positive donor cytomegalovirus serology, longer graft ischemic time, and lower donor body weight. Several interactions between individual donor characteristics affected survival. In male donors, history of hypertension and diabetes mellitus were risk factors for death (p = 0.006, p = 0.04, respectively), but not in female donors (p = 0.5, p = 0.8, respectively). There was a significant interaction between donor age and recipient-donor weight difference. If the donor was of younger age, increasing recipient-donor weight difference did not result in increased death. With increasing donor age, weight difference did result in compromised survival (p < 0.0003). Donor and recipient gender further modified the degree of risk: risk was higher in female donors and when recipients were male (p < 0.0003). CONCLUSIONS: This multi-institutional analysis identified important interactions between donor characteristics that affect post-transplant survival that explain some of the discrepancies in the results of previous studies. The results are likely to aid in efficient organ allocation. PMID- 19804990 TI - Pericardial constriction after cardiac transplantation. AB - In this study we present a series of 5 cases that developed constrictive pericarditis after orthotopic heart transplantation. All 5 patients had pericardial effusion of non-infectious etiology in the early post-transplant period. They subsequently presented with heart failure unresponsive to standard medical management. The diagnosis was made by comprehensive echo-Doppler studies. Findings were confirmed at surgical inspection and complete pericardiectomy led to improvement in hemodynamics in 4 patients. One patient had relief from constriction but died of non-cardiac complications. One patient with constriction has been re-listed for transplantation due to intermittent heart block and associated cardiac allograft vasculopathy. Early diagnosis of pericardial constriction after orthotopic heart transplantation requires a high index of clinical suspicion and optimal use of Doppler echocardiography. Early diagnosis and timely surgical pericardiectomy may correct this condition entirely and result in satisfactory long-term results. PMID- 19804991 TI - Testing the repression hypothesis: effects of emotional valence on memory suppression in the think - no think task. AB - It has been proposed that performance in the think - no think (TNT) task represents a laboratory analogue of the voluntary form of memory repression. The central prediction of this repression hypothesis is that performance in the TNT task will be influenced by emotional characteristics of the material to be remembered. This prediction was tested in two experiments by asking participants to learn paired associates in which the first item was either emotionally positive (e.g. joy) or emotionally negative (e.g. hatred). The second word was always emotionally neutral (e.g. socks). Consistent with the repression hypothesis, significant memory suppression was observed in both experiments following 'no think' instructions for memories associated with emotionally negative material. No suppression was observed for memories associated with emotionally positive information. Implications of these findings for the relationship between performance in the TNT task and the controversial notion of memory repression are considered. PMID- 19804992 TI - Forms, functions, and foibles of humor used in AIDS service organizations. AB - Research has indicated that HIV service providers commonly use humor to cope with work-related stress; however, little is known about the forms and functions of humor used by these professionals. In this study, 25 HIV service providers from five AIDS service organizations were interviewed about their use of humor. Participants described five primary types of humor as prevalent within AIDS service organizations and noted that humor served a variety of functions, which were either adaptive or maladaptive. Adaptive functions included boosting morale and reducing tension, whereas maladaptive functions ranged from masking emotions to alienating certain groups. Results emphasized the importance of context in the study of humor use and the need for continued investigations of the stress and coping of HIV service providers. PMID- 19804993 TI - Protective factors for HIV infection among Mexican American men who have sex with men. AB - Latinos in the United States have been disproportionately affected by the HIV epidemic. The purpose of this study was to identify potential themes for inclusion in effective HIV prevention interventions for Mexican American men who have sex with men (MSM). The authors used a phenomenological design to explore the lived experiences of Mexican American MSM who had grown up in Dallas, Texas, regarding protective factors for HIV infection. A total of 20 30- to 60-year-old Mexican American MSM participated in semistructured interviews. During data analysis, the following themes concerning protective behaviors for HIV emerged: (a) accepting one's sexuality; (b) machismo; (c) being in love; (d) respect for family, self, and life; and (e) having HIV-living now. Strategies for potential inclusion in HIV prevention interventions geared toward Mexican American MSM were identified based on these themes. The recommendations encompass modification of behavioral interventions and related social policies. PMID- 19804994 TI - An examination of the psychometric properties of the Antiretroviral General Adherence Scale (AGAS) in two samples of HIV-infected individuals. AB - This article reports on the development and psychometric properties of the Antiretroviral General Adherence Scale (AGAS) in two National Institutes of Health-funded projects: the Get Busy Living Project, a behavioral clinical trial to promote consistent use of antiretroviral therapy, and the KHARMA (Keeping Healthy and Active with Risk Reduction and Medication Adherence) Project, which addressed issues of adherence and risk reduction behavior in women. AGAS assesses the ease and ability of participants to take antiretroviral therapy according to a health care provider's recommendations. Data were analyzed from completed baseline assessments of the two studies. The AGAS was internally consistent in both samples. Content, construct, and criterion validity were established using factor analysis and correlations of total AGAS scores with two measures of adherence: electronic drug monitoring and an Adult AIDS Clinical Trials Group adherence scale. Viral load, CD4 cell counts, and depression scores were also examined. Reliability and validity of the AGAS were supported in both samples. PMID- 19804995 TI - Smoking cessation for persons living with HIV: a review of currently available interventions. AB - Smoking among persons living with HIV infection (PLWH) is estimated to be two to three times greater than the general population. Data suggest that cigarette smoking is more common among PLWH because of factors such as lower socioeconomic status, previous or concurrent illicit drug and alcohol use, age, education level, and concurrent depressive symptoms. Cigarette smoking in HIV-infected individuals has been associated with higher risk of certain cancers, infections, and a decrease in response to antiretroviral therapy. Recent guidelines from the U.S. Public Health Service have recommended the use of counseling and pharmacologic therapies for all patients interested in quitting smoking. It is critical for clinicians caring for PLWH to be familiar with these modalities. This article provides an overview of currently available interventions for smoking cessation, reviews considerations for patients with HIV, and discusses clinical implications for nurses and advanced practice clinicians. PMID- 19804996 TI - Force majeure: therapeutic measures in response to restricted supply of imiglucerase (Cerezyme) for patients with Gaucher disease. AB - Gaucher disease is the first lysosomal disorder for which clinically effective enzyme replacement therapy has been introduced. Lifelong treatment with imiglucerase, the recombinant glucocerebrosidase manufactured by the Genzyme Corporation (MA, USA), is administered intravenously - usually at biweekly intervals. An acute shortage of imiglucerase (to 20% of prior global supply) has occurred as a result of viral contamination of the production facility; production was halted, and a full supply of imiglucerase is not anticipated until January 2010. An urgent meeting of physicians, researchers, and patients was convened through the agency of the European Working Group for Gaucher Disease; this was instigated by patients internationally represented by the European Gaucher Alliance. Here we present a position statement based on the findings of the group, with key recommendations about identification and monitoring of at risk patients threatened by the abrupt withdrawal of treatment, the equitable distribution of residual imiglucerase - and access to alternative treatments including those that have completed phase III clinical trials but have not yet been licensed. PMID- 19804997 TI - Tail-biting: a new perspective. AB - Tail-biting data from different studies are difficult to compare because a range of definitions of tail-biting behaviour and tail-biting lesions are used. Although records from abattoirs provide a large database, their usefulness is restricted as tail-biting is under-recorded and environmental and husbandry factors associated with the behaviour are unlikely to be known. Both farm and abattoir data provide no information on the number of pigs biting, only those bitten. Studying individual animals that tail-bite should give a better understanding of the pig's motivation to tail-bite and which of the components of its environment should be adjusted to improve welfare. This review examines the existing literature on tail-biting in pigs but considered from a new perspective using three different descriptive behavioural types, namely, 'two-stage', 'sudden forceful' and 'obsessive', each of which may have different motivational bases. The article also considers the different environmental and husbandry factors which may affect each type of behaviour and discusses why this is such a complicated field and why it is often difficult to draw conclusions from available research. PMID- 19804998 TI - Effect of diet, energy balance and milk production on oxidative stress in early lactating dairy cows grazing pasture. AB - The aim of this study was to determine the effect of diet, energy balance and milk production on oxidative stress in early-lactating, Holstein-Friesian dairy cows fed to produce either low or high levels of milk. Indicators of energy balance (non-esterified fatty acids, beta-hydroxybutyrate, glucose and insulin like growth factor-1) and indicators of oxidative stress (reactive oxygen metabolites and biological antioxidants) were measured in the first 5 weeks of lactation. Energy balance indicators showed that high producing animals had a lower degree of negative energy balance. Diet was found to have an indirect effect on the level of oxidative stress. Factors associated with a high level of oxidative stress were severe negative energy balance (mean -71 +/- 6.85 27 MJ/cow/day, P < 0.05) and lower levels of milk production (mean 26.4 +/- 0.07 28 L/cow/day, P < 0 .05). Further studies will be required to more precisely determine the specific effects of diet, energy balance and milk production on such stress in dairy cows and to establish normal ranges for these biomarkers. PMID- 19804999 TI - An approach to automated frequency-domain feature extraction in nuclear magnetic resonance spectroscopy. AB - For the analysis of metabolite systems, nuclear magnetic resonance (NMR) spectroscopy has become an important quantitative monitoring technology. Automated quantitation methods are highly desired and mainly characterized by the tasks of model selection and parameter approximation. This paper proposes a promising automated two stage approach in the frequency-domain, in which signaling peaks are first identified and filtered from noise based on curvature properties of the spectrum, and then proportionally approximated based on the analytical solution of a Lorentz-function. Remarkably, in opposition to common least-squares approaches, the proposed approximation scheme does not rely on partial derivatives, and furthermore, the runtime is independent to the number of spectral datapoints. Simulations provide promising empirical evidence for successful peak selection and parameter approximation, with the results for the latter highly outperforming the Levenberg-Marquardt algorithm in terms of error minimization and robustness. PMID- 19805000 TI - Systematic review of apomorphine infusion, levodopa infusion and deep brain stimulation in advanced Parkinson's disease. AB - The effectiveness of oral levodopa in complex Parkinson's disease (PD) is limited by its short half-life, and the resulting pulsatile dopaminergic stimulation leads to complex motor fluctuations and dyskinesia. Several treatments provide more continuous/less pulsatile dopaminergic stimulation by modifying the pharmacokinetics of levodopa or dopamine; however, patients with advanced disease can be refractory to these treatments. In such cases infusion therapies (apomorphine and intraduodenal levodopa) and neurosurgery (deep brain stimulation [DBS]) may be used. The purpose of this systematic review is to assess, as far as possible, the relative effectiveness of these therapies. There were no randomised controlled trials comparing the three treatment modalities or any directly comparable studies, therefore a descriptive analysis of the data was performed. Studies identified for levodopa infusion and DBS supported a significant benefit compared with best medical management in terms of improvements in the proportion of the waking day in a functional "on" state, activities of daily living and motor score. This finding was supported in observational studies for all three therapies. Adverse events were not adequately reported in the majority of included studies and it was therefore not possible to obtain a reliable tolerability profile of the different treatment options. The absence of direct comparative data means that, for the immediate future at least, treatment choices for advanced PD will be determined by clinical judgement and patient preference. There is an urgent need for well-designed clinical trials to generate reliable data to inform the clinical management of this difficult-to-treat subgroup of PD patients. PMID- 19805001 TI - Effect of weathering and thickness on the superficial microhardness of acrylic resin and ocular button. AB - PURPOSE: This study evaluated the effects of weathering (1008h) and thickness (1 and 3.5mm) on the superficial microhardness of the ocular button, using five colorless acrylic resins, and colorless buttons specific for ocular prosthesis. MATERIALS AND METHODS: One-hundred-and-twenty specimens were made and distributed allocated to 12 groups. Microhardness analysis was carried out with a microdurometer (Shimadzu, Japan) before and after weathering for 1008h. Data were then analyzed statistically by ANOVA and the Tukey Test at 1% significance. RESULTS: The data demonstrated that the ocular button and the QC 20 resin presented, respectively, the highest (22.9) and lowest (18.31) values of surface microhardness (Knoop), independently of the variable evaluated. CONCLUSION: Thickness did not influence the property studied, and the specimens presented higher mean values of superficial microhardness after weathering for 1008h. PMID- 19805002 TI - Recent advances in mycobacterial cell wall glycan biosynthesis. AB - The cell wall of mycobacteria, including the causative agents of the human diseases tuberculosis (Mycobacterium tuberculosis) and leprosy (M. leprae), is composed of an array of carbohydrate-containing molecules. These glycoconjugates are assembled by glycosyltransferases (GTs) that work in tandem through pathways that are only now beginning to be fully understood. Given the essentiality of cell wall glycans to mycobacterial viability, these enzymes represent novel targets for drug action. Summarized here are recent genetic and biochemical studies leading to the identification and characterization of mycobacterial GTs. PMID- 19805004 TI - Direct genotyping of cytomegalovirus envelope glycoproteins from toddler's saliva samples. AB - BACKGROUND: The polymorphism of genes encoding CMV envelope protein is used for strain classification and may influence pathogenesis and/or infectivity. CMV genotyping is usually based on sequencing or acrylamide gel-RFLP, but these methods are not suited to rapid screening of large populations. OBJECTIVES: We developed a high-throughput method to analyze CMV strains diversity and to detect multiple-strain infection in a large population of toddlers (six daycare centers (DCC) and an emergency unit (EU)). METHODS: We developed a new PCR-RFLP method coupled with capillary electrophoresis fragment detection for UL55-gB, UL75-gH and UL73-gN genotyping. To detect gB recombinants, gpUL55 typing was applied to two variable zones (NTerminal and central). We applied this method to 212 CMV positive saliva samples and controlled the results by direct sequencing of PCR products. RESULTS: We identified 112 strains, that fell into eight groups in UL55 gB, two groups in UL75-gH, and seven groups in UL73-gN. The 79 samples from the emergency unit contained 30 strains, 28 children harboring 2 strains. The samples (n=133) from the six daycare centers contained respectively 4, 1, 6, 1 and 11 strains. Fifteen percent of strains were UL55-gB recombinants. CONCLUSION: Our new method can simultaneously determine gB, gH and gN genotypes and offers more precise classification of CMV strains than previous RFLP-based methods. This could constitute the basis for a new classification, particularly in UL55-gB. Easy direct identification of multiple strains and recombinants in pathological samples could facilitate large epidemiologic studies. PMID- 19805003 TI - Novel strategies for reversing platinum resistance. AB - Platinum-based drugs continue to be the mainstay of therapy for ovarian cancer. Along with adverse effects, chemoresistance (intrinsic or acquired) has become a major limitation in the management of recurrent disease. Even though much is known about the effects of platinum drugs on cancer cells, the mechanisms underlying resistance are poorly understood. In this review, we summarize the current data on chemoresistance and discuss novel strategies to reverse resistance to platinum-based drugs. The most important targets highlighted here include Aurora kinases, PARP, ATP7B, and ERCC1. Furthermore, we discuss the implications of these novel approaches for ovarian cancer treatment. PMID- 19805005 TI - Staphylococcal lipoproteins and their role in bacterial survival in mice. AB - Staphylococcus aureus expresses about 50 lipoproteins (Lpp), which are lipid anchored in the membrane. The processing of the precursor to the mature Lpp is catalyzed by the phosphatidyl glycerol diacylglyceryl transferase (Lgt) and the lipoprotein-specific type II signal peptidase (LspA) leading to diacylated Lpp. Possibly another acyltransferase attaches a third fatty acid leading to triacylated Lpp. Lpp function as binding proteins for transport of nutrients across the microbial membrane and are involved in processing of other proteins, but most Lpp remain of predicted or unknown function. The di- or triacylated lipid structure is sensed by host pattern recognition receptor TLR2 and induces innate immune responses in professional and non-professional phagocytes. In the host, maturation of Lpp confers optimal metal ion - particularly iron - acquisition, it enhances staphylococcal invasion and phagocytosis, intracellular survival and persistence of infections. However, the advantages of Lpp maturation are counterbalanced by the capability to induce inflammation. In this review, we summarize the current knowledge about the role of Lpp in iron acquisition and TLR2 recognition in the host and describe the consequences of Lpp maturation for survival of S. aureus in the host. PMID- 19805006 TI - A census of the advanced and specialist cancer nursing workforce in England, Northern Ireland and Wales. AB - INTRODUCTION: At present no detailed data on the specialist cancer nursing workforce across different cancer types and populations is routinely collected in England, Wales or Northern Ireland. This has implications for workforce planning and the future provision of cancer services. METHOD: In an attempt to establish a baseline of the workforce a census was taken. Data was collected via an Excel spreadsheet by Cancer Network Nurse Directors and Lead Nurses across England and Northern Ireland and a lead nurse in Wales. Scotland, Palliative Care and Chemotherapy posts were excluded at levels other than Consultant as these are collected via other mechanisms. RESULTS & CONCLUSIONS: The census recorded 2309.4 specialist and advanced practice posts in England (89% response rate), 204 posts in Wales (66% response rate) and 43.4 posts in Northern Ireland (100% response rate). There is a variation in terms of distribution of specialist nurses across the Networks both in number and in cancer type. 1800 adult CNS posts were recorded in England and 1 in 5 of these were breast cancer posts. The range of job titles is very wide with 17 different titles being used. In England the extent of support for posts from Macmillan Cancer Support was considerable. Around a third (31%) of all adult cancer specialist posts in England are supported by Macmillan Cancer Support (n=671.2) 607.2 of these are CNS posts equating to 34% of all CNS posts in England. 34% of all NI CNS posts and 32.5% in Wales. PMID- 19805007 TI - The management of women with abnormal cervical cytology in pregnancy. AB - The management of women with abnormal cytology in pregnancy represents both a diagnostic and a therapeutic challenge for colposcopists. The emphasis should be on diagnosis and confirmation of cervical precancer (Cervical intraepithelial neoplasia (CIN) or Adenocarcinoma in situ (AIS), thus excluding invasive cancer). Following an initial assessment, careful follow-up is essential. This must include colposcopy and take into account the physiological changes of the cervix during pregnancy and the puerperium. The management of women with invasive cancer diagnosed during pregnancy depends on the gestation at diagnosis and requires careful assessment and multidisciplinary planning. PMID- 19805008 TI - Simultaneous analysis of eight nucleoside triphosphates in cell lines by liquid chromatography coupled with tandem mass spectrometry. AB - In this study, we developed a new method for the simultaneous determination of eight endogenous ribonucleoside triphosphates and deoxyribonucleoside triphosphates based on a combination of a selective sample preparation and an ion pair liquid chromatography-electrospray tandem mass spectrometry. The sample preparation was based on a protein precipitation coupled with a solid phase extraction using a weak-anion-exchange cartridge. The analytical separation of the nucleotides was achieved on a porous graphitic carbon stationary phase with a binary elution gradient program employing ion-pairing reagents (diethylamine and hexylamine) and organic eluent (methanol). The triple quadrupole mass spectrometer operated in both negative and positive multiple reaction monitoring modes. The calibration assay used the stable isotope labelled analogs of each compounds as standard. Standard calibrations were from 0.25 to 10pmol injected according to deoxyribonucleotides and from 12.5 to 3000pmol injected according to ribonucleotides. The within-run precision of the assay was less than 14.5% and the between-run precision was less than 12.4% for each analytes. Assay accuracy was in the range of 92.3-107.6%. This method allows the determination of NTP and dNTP pools from lysats of several cell lines or peripheral blood mononuclear cell from patient. Assays were performed with different preparation of cells to confirm the quality and the relevance of the described method. PMID- 19805009 TI - Autoimmune pancreatitis: a challenging diagnostic puzzle for clinicians. AB - Autoimmune pancreatitis is a form of pancreatitis with autoimmune stigmata that may present as either focal or diffuse gland involvement. In focal forms, autoimmune pancreatitis shares demographic, clinical, biochemical and imaging features with pancreatic cancer. Since autoimmune pancreatitis is a benign disease and steroid therapy can rapidly resolve symptoms, improve radiological findings and avoid unnecessary surgery, the current clinical challenge is how to differentiate autoimmune pancreatitis from pancreatic neoplasia. Even though definitive diagnosis of the disease is difficult, several diagnostic criteria have been proposed and progress has been made in imaging studies. The management of this unique form of pancreatitis should, therefore, be handled in centres with knowledge of all aspects of the disease. This article briefly reviews clinical aspects of autoimmune pancreatitis with a focus on its diagnostic imaging and management. PMID- 19805010 TI - [Star anise poisoning]. PMID- 19805011 TI - [Acute necrotising pneumonia eighteen months after aspiration of a pine branch]. PMID- 19805012 TI - Aesthetic outcome and oncological safety of nipple-areola complex replantation after mastectomy and immediate breast reconstruction. AB - Immediate breast reconstruction (IBR) has become an established procedure for women necessitating mastectomy. Traditionally, the nipple-areola complex (NAC) is resected during this procedure. The NAC, in turn, is a principal factor determining aesthetic outcome after breast reconstruction, and due to its particular texture and shape, a natural-looking NAC can barely be reconstructed with other tissues. The aim of this study was to assess the oncological safety as well as morbidity and aesthetic outcome after replantation of the NAC some days after IBR. Retrospective analysis of 85 patients receiving 88 mastectomies and IBR between 1998 and 2007 was conducted. NAC (n=29) or the nipple alone (n=23) were replanted 7 days (median, range 2-10 days) after IBR in 49 patients, provided the subareolar tissue was histologically negative for tumour infiltration. Local recurrence rate was assessed after 49 months (median, range 6 120 months). Aesthetic outcome was evaluated by clinical assessment during routine follow-up at least 12 months after the last intervention. Malignant involvement of the subareolar tissue was found in eight cases (9.1%). Patients qualifying for NAC replantation were in stage 0 in 29%, stage I in 15%, stage IIa in 31%, stage IIb in 17% and stage III in 8%. Total or partial necrosis occurred in 69% and 26% if the entire NAC or only the nipple were replanted, respectively (P<0.01). Depigmentation was seen in 52% and corrective surgery was done in 11 out of 52 NAC or nipple replantations. Local recurrence and isolated regional lymph node metastasis were observed in one single case each. Another 5.8% of the patients showed distant metastases. We conclude that the replantation of the NAC in IBR is oncologically safe, provided the subareolar tissue is free of tumour. However, the long-term aesthetic outcome of NAC replantation is not satisfying, which advocates replanting the nipple alone. PMID- 19805013 TI - A survey of treatment practices in diabetic peripheral neuropathy. AB - OBJECTIVE: To evaluate provider practices for identification and treatment of painful diabetic peripheral neuropathy (DPN). METHODS: A questionnaire was distributed to healthcare providers attending educational programs in New England. RESULTS: The survey was completed by 357 providers. Although generally ineffective in neuropathic pain, non-steroidal anti-inflammatory drugs were prescribed by 31% of providers. Only 57% providers used a quantitative pain scale to evaluate pain. The effectiveness of medications was assessed at least frequently by 70% of providers and at every visit by 22% providers. CONCLUSION: The results reiterate the need for routinely monitoring patients with painful DPN and using appropriate pain scales. PMID- 19805014 TI - Antenatal screening and predicting hypertension in pregnancy for midwives. AB - BACKGROUND: The cause of hypertension in pregnancy remains unknown and results in increased risk of complications for mother and baby. Symptoms of developing pre eclampsia, such as an elevated blood pressure, can be vague and singular. The purpose of this literature review is to evaluate research investigating antenatal screening practices for hypertension which fall within the midwives scope of practice. METHOD: Inclusion criteria for this literature review were English language, peer reviewed primary research journal articles, published in the previous 20 years where the population under study was pregnant with reported outcomes of prevention, screening or prediction of hypertension in pregnancy. A large number of papers (n=201) were identified and these were screened and subsequently excluded if they addressed diagnostic testing, screening and interpretation that depended solely on a medical practitioner. RESULTS: There was no single predictive factor found, however the relevant papers included in this review (n=33) found evidence of modifiable, non-modifiable and clinical assessment factors for inclusion in a midwifery screening model. CONCLUSIONS: Further research should be focused on the factors observed by midwives during history taking and the antenatal course in the second and third trimesters and whether or not these can be synthesised in to a hypertension-specific diagnostic tool for use in midwifery practice. PMID- 19805015 TI - Vaccine development against Trypanosoma cruzi and Leishmania species in the post genomic era. AB - Trypanosoma cruzi and the genus Leishmania are protozoan parasites causing diseases of major public health importance, and the recent completion of the sequencing of their genomes has opened new opportunities to further our understanding of the mechanisms required for protection and the development of vaccines. For example, trans-sialidases, one of the largest protein families from T. cruzi, contain dominant CD8+ T cell epitopes, and their use as preventive or therapeutic vaccines in different animal models has provided encouraging results. A much wider range of antigens and vaccine formulations have been tested against Leishmania, and new correlates for protection are being defined, such as the induction of multifunctional Th1 effector cells capable of producing a complex set of cytokines. Also, while a large number of these vaccine candidates have been rather successful in mouse models, their usefulness in more relevant animal models is still poor, in spite of significant immunogenicity. Novel proteomics and genomics approaches are being used for antigen discovery and the identification of new vaccine candidates, some of which have shown promise for the control of infection. These studies cast little doubt that T. cruzi and Leishmania genomes represent major resources for understanding key aspects of the mechanisms of immune protection against these parasites, and the increasing use of these tools will greatly impact vaccine development. PMID- 19805016 TI - Estimates of health care system costs of unsafe abortion in Africa and Latin America. AB - CONTEXT: Each year, 19 million unsafe abortions occur in developing countries, and an estimated five million women are treated for the resulting serious medical complications. Meanwhile, the economic impact of postabortion care on health care systems in Africa and Latin America is poorly understood (data for Asia are lacking). METHODS: Two main approaches were used to estimate the cost of postabortion care: calculating the average cost of care per patient, as represented in 20 empirical studies, and analyzing treatment costs using the WHO Mother-Baby Package model, which enumerates the costs of specific components of treatment related to postabortion complications. The average cost estimates from each approach were multiplied by the annual number of cases of hospitalization for postabortion care to generate regional cost estimates. Three methods (low severity, weighted severity, and inclusion of overhead and capital costs) were used to generate a range of per-patient and regional cost estimates. RESULTS: The average per-patient cost of postabortion care ranged from $83 in Africa to $94 in Latin America (2006 US$); estimates based on the WHO Mother-Baby Package model were between $57 and $109 per case. The health system costs of postabortion care in the two regions combined ranged from $159 million to $333 million per year. The average estimates from the two approaches were similar: $280 million and $274 million, respectively. CONCLUSIONS: The costs of treating medical complications from unsafe abortion constitute a significant financial burden on public health care systems in the developing world, and postabortion complications are a significant cause of maternal morbidity. PMID- 19805017 TI - Demand and unmet need for means of family limitation in Rwanda. AB - CONTEXT: Rwanda is the most densely populated country in Africa, with substantial annual population growth. The current government seeks new policies for family limitation as a way to facilitate more sustainable development. METHODS: Data from the 2005 Rwanda Demographic and Health Survey were used for a two-step analysis; binary logistic regression was used to identify factors associated with desiring to stop childbearing and having unmet need. RESULTS: Eighty-seven percent of women aged 15-49 approve of family planning, but only 64% believe that their partner approves of it. There is a high level of unmet need for family limitation; 58% of women who want to stop childbearing do not use modern contraceptives. Demand was lower among women who did not approve of family planning, those who did not know their partner's attitude toward family planning and those who had discussed family planning with their partner fewer than three times. Unmet need was higher among women who did not approve of family planning, those who believed their partner did not approve of family planning or who did not know his attitude, and those who had never discussed family planning with their partner or had done so only once or twice. CONCLUSIONS: Negative attitudes toward family planning and failing structures of provision are the dominant constraints on the use of modern contraceptives in Rwanda. Community-based family planning services could greatly expand access, especially in underserved provinces. PMID- 19805018 TI - Male perceptions on female sterilization: a community-based study in rural Central India. AB - CONTEXT: Use of modern contraceptive methods has increased fourfold in India since the 1970s, characterized by a predominance of female sterilization. There has been considerable investigation about women's choice of female sterilization, but little from the male perspective. METHODS: Seven focus group discussions were conducted among 58 men currently married to women aged 15-45, followed by a cross sectional survey among 793 men currently married to same-aged women. Bivariate analysis was used for the survey data, and content analysis was used for the qualitative data. RESULTS: Men's primary source of reproductive health information was mass media, although they expressed interest in getting information through discussion with knowledgeable sources. Men understood family planning and contraception to be two separate issues: Men viewed "family planning" as synonymous with female sterilization, whereas they saw "contraception" as referring to spacing methods, knowledge of which was limited. Thirty-four percent of men reported that their wives had been sterilized; 79% of men who did not rely on any permanent method said they wanted their wives to be sterilized. In focus group discussions, most men reported themselves as their family's sole decision maker about reproductive health; however, only one-third of survey respondents did so. CONCLUSION: Men are interested in acquiring family planning information, but lack knowledge about available information sources, which hampers their ability to make informed family planning choices. Family planning service providers and program planners need to be aware of males' knowledge and perceptions pertaining to family planning, and make appropriate modifications to communication strategies. PMID- 19805019 TI - The role of education level in the intergenerational pattern of adolescent pregnancy in Brazil. AB - CONTEXT: Adolescent pregnancy has been associated with the early childbearing experience of the mothers of adolescents, and young people's education level is believed to be an important factor in this phenomenon. METHODS: In 2002, a representative household survey collected data from 3,050 young men and women aged 20-24 in three Brazilian cities. The main measures were mother's age at first birth, daughter's age at first pregnancy and son's age when he first impregnated a partner; ages were dichotomized as younger than 20 and 20 or older. The distribution of respondents by both their own and their mothers' reproductive experience was analyzed in relation to various characteristics, and logistic regressions assessed possible associations between these variables and pregnancy experience. RESULTS: Thirty percent of women reported getting pregnant before age 20, and 21% of men said they were younger than 20 when they first impregnated a partner. Of these groups, 34% of women and 31% of men reported that their mothers had first given birth at the same age. Both women and men were more likely to have had an early pregnancy experience if their mother had had a child before age 20 (odds ratios, 2.0 and 2.3, respectively). Among women, this positive association disappeared in the final model after adjusting for their education level, whereas among men the association remained after similar adjustment (1.8). CONCLUSIONS: Daughters' and sons' level of education appears to be an important factor in the repetition of adolescent fertility across generations. Efforts are needed to increase access to education and to encourage young people to remain in school. PMID- 19805020 TI - Evidence-based, alternative cervical cancer screening approaches in low-resource settings. PMID- 19805021 TI - Experimental and clinical evidence of neuroprotection by nerve growth factor eye drops: Implications for glaucoma. AB - Elevated intraocular pressure (IOP) in glaucoma causes loss of retinal ganglion cells (RGCs) and damage to the optic nerve. Although IOP is controlled pharmacologically, no treatment is available to restore retinal and optic nerve function. We evaluated the effects of NGF eye drops in a rat model of glaucoma. We also treated 3 patients with progressive visual field defects despite IOP control. Glaucoma was induced in rats through injection of hypertonic saline into the episcleral vein. Initially, 2 doses of NGF (100 and 200 mug/mL) were tested on 24 rats, and the higher dose was found to be more effective. Glaucoma was then induced in an additional 36 rats: half untreated and half treated with 200 mug/mL NGF QID for 7 weeks. Apoptosis/survival of RGCs was evaluated by histological, biochemical, and molecular analysis. Three patients with advanced glaucoma underwent psychofunctional and electrofunctional tests at baseline, after 3 months of NGF eye drops, and after 3 months of follow-up. Seven weeks of elevated IOP caused RGC degeneration resulting in 40% cell death. Significantly less RGC loss was observed with NGF treatment (2,530 +/- 121 vs. 1,850 +/- 156 RGCs/mm(2)) associated with inhibition of cell death by apoptosis. Patients treated with NGF demonstrated long lasting improvements in visual field, optic nerve function, contrast sensitivity, and visual acuity. NGF exerted neuroprotective effects, inhibiting apoptosis of RGCs in animals with glaucoma. In 3 patients with advanced glaucoma, treatment with topical NGF improved all parameters of visual function. These results may open therapeutic perspectives for glaucoma and other neurodegenerative diseases. PMID- 19805022 TI - Type 2C protein phosphatases directly regulate abscisic acid-activated protein kinases in Arabidopsis. AB - Abscisic acid (ABA) signaling is important for stress responses and developmental processes in plants. A subgroup of protein phosphatase 2C (group A PP2C) or SNF1 related protein kinase 2 (subclass III SnRK2) have been known as major negative or positive regulators of ABA signaling, respectively. Here, we demonstrate the physical and functional linkage between these two major signaling factors. Group A PP2Cs interacted physically with SnRK2s in various combinations, and efficiently inactivated ABA-activated SnRK2s via dephosphorylation of multiple Ser/Thr residues in the activation loop. This step was suppressed by the RCAR/PYR ABA receptors in response to ABA. However the abi1-1 mutated PP2C did not respond to the receptors and constitutively inactivated SnRK2. Our results demonstrate that group A PP2Cs act as 'gatekeepers' of subclass III SnRK2s, unraveling an important regulatory mechanism of ABA signaling. PMID- 19805023 TI - Rapid equilibrium sampling initiated from nonequilibrium data. AB - Simulating the conformational dynamics of biomolecules is extremely difficult due to the rugged nature of their free energy landscapes and multiple long-lived, or metastable, states. Generalized ensemble (GE) algorithms, which have become popular in recent years, attempt to facilitate crossing between states at low temperatures by inducing a random walk in temperature space. Enthalpic barriers may be crossed more easily at high temperatures; however, entropic barriers will become more significant. This poses a problem because the dominant barriers to conformational change are entropic for many biological systems, such as the short RNA hairpin studied here. We present a new efficient algorithm for conformational sampling, called the adaptive seeding method (ASM), which uses nonequilibrium GE simulations to identify the metastable states, and seeds short simulations at constant temperature from each of them to quantitatively determine their equilibrium populations. Thus, the ASM takes advantage of the broad sampling possible with GE algorithms but generally crosses entropic barriers more efficiently during the seeding simulations at low temperature. We show that only local equilibrium is necessary for ASM, so very short seeding simulations may be used. Moreover, the ASM may be used to recover equilibrium properties from existing datasets that failed to converge, and is well suited to running on modern computer clusters. PMID- 19805024 TI - BAC talk about cell type-specific regulation of human IL-10. PMID- 19805025 TI - CIB1 functions as a Ca(2+)-sensitive modulator of stress-induced signaling by targeting ASK1. AB - Calcium and integrin binding protein 1 (CIB1) is a Ca(2+)-binding protein of 22 kDa that was initially identified as a protein that interacts with integrin alpha(IIb). Although it interacts with various proteins and has been implicated in diverse cellular functions, the molecular mechanism by which CIB1 regulates intracellular signaling networks has remained unclear. We now show that, by targeting apoptosis signal-regulating kinase 1 (ASK1), CIB1 negatively regulates stress-activated MAPK signaling pathways. CIB1 was thus shown to bind to ASK1, to interfere with the recruitment of TRAF2 to ASK1, and to inhibit the autophosphorylation of ASK1 on threonine-838, thereby blocking ASK1 activation. Furthermore, CIB1 mitigated apoptotic cell death initiated either by TNF-alpha in breast cancer MCF7 cells or by 6-hydroxydopamine (6-OHDA) in dopaminergic cells. Ca(2+) influx induced by membrane depolarization reversed the inhibitory effect of CIB1 on 6-OHDA-induced ASK1 activation and cell death in dopaminergic neurons. These observations thus suggest that CIB1 functions as a Ca(2+)-sensitive negative regulator of ASK1-mediated signaling events. PMID- 19805026 TI - Some cautionary notes on fisheries evolutionary impact assessments. PMID- 19805027 TI - Do mangroves provide an effective barrier to storm surges? PMID- 19805028 TI - Structural organization of Weibel-Palade bodies revealed by cryo-EM of vitrified endothelial cells. AB - In endothelial cells, the multifunctional blood glycoprotein von Willebrand Factor (VWF) is stored for rapid exocytic release in specialized secretory granules called Weibel-Palade bodies (WPBs). Electron cryomicroscopy at the thin periphery of whole, vitrified human umbilical vein endothelial cells (HUVECs) is used to directly image WPBs and their interaction with a 3D network of closely apposed membranous organelles, membrane tubules, and filaments. Fourier analysis of images and tomographic reconstruction show that VWF is packaged as a helix in WPBs. The helical signature of VWF tubules is used to identify VWF-containing organelles and characterize their paracrystalline order in low dose images. We build a 3D model of a WPB in which individual VWF helices can bend, but in which the paracrystalline packing of VWF tubules, closely wrapped by the WPB membrane, is associated with the rod-like morphology of the granules. PMID- 19805029 TI - CAPS drives trans-SNARE complex formation and membrane fusion through syntaxin interactions. AB - Ca(2+)-dependent activator protein for secretion (CAPS) is an essential factor for regulated vesicle exocytosis that functions in priming reactions before Ca(2+)-triggered fusion of vesicles with the plasma membrane. However, the precise events that CAPS regulates to promote vesicle fusion are unclear. In the current work, we reconstituted CAPS function in a SNARE-dependent liposome fusion assay using VAMP2-containing donor and syntaxin-1/SNAP-25-containing acceptor liposomes. The CAPS stimulation of fusion required PI(4,5)P(2) in acceptor liposomes and was independent of Ca(2+), but Ca(2+) dependence was restored by inclusion of synaptotagmin. CAPS stimulated trans-SNARE complex formation concomitant with the stimulation of full membrane fusion at physiological SNARE densities. CAPS bound syntaxin-1, and CAPS truncations that competitively inhibited syntaxin-1 binding also inhibited CAPS-dependent fusion. The results revealed an unexpected activity of a priming protein to accelerate fusion by efficiently promoting trans-SNARE complex formation. CAPS may function in priming by organizing SNARE complexes on the plasma membrane. PMID- 19805030 TI - SH3TC2/KIAA1985 protein is required for proper myelination and the integrity of the node of Ranvier in the peripheral nervous system. AB - Charcot-Marie-Tooth disease type 4C (CMT4C) is an early-onset, autosomal recessive form of demyelinating neuropathy. The clinical manifestations include progressive scoliosis, delayed age of walking, muscular atrophy, distal weakness, and reduced nerve conduction velocity. The gene mutated in CMT4C disease, SH3TC2/KIAA1985, was recently identified; however, the function of the protein it encodes remains unknown. We have generated knockout mice where the first exon of the Sh3tc2 gene is replaced with an enhanced GFP cassette. The Sh3tc2(DeltaEx1/DeltaEx1) knockout animals develop progressive peripheral neuropathy manifested by decreased motor and sensory nerve conduction velocity and hypomyelination. We show that Sh3tc2 is specifically expressed in Schwann cells and localizes to the plasma membrane and to the perinuclear endocytic recycling compartment, concordant with its possible function in myelination and/or in regions of axoglial interactions. Concomitantly, transcriptional profiling performed on the endoneurial compartment of peripheral nerves isolated from control and Sh3tc2(DeltaEx1/DeltaEx1) animals uncovered changes in transcripts encoding genes involved in myelination and cell adhesion. Finally, detailed analyses of the structures composed of compact and noncompact myelin in the peripheral nerve of Sh3tc2(DeltaEx1/DeltaEx1) animals revealed abnormal organization of the node of Ranvier, a phenotype that we confirmed in CMT4C patient nerve biopsies. The generated Sh3tc2 knockout mice thus present a reliable model of CMT4C neuropathy that was instrumental in establishing a role for Sh3tc2 in myelination and in the integrity of the node of Ranvier, a morphological phenotype that can be used as an additional CMT4C diagnostic marker. PMID- 19805031 TI - Toll-like receptor 2 dependent immunogenicity of glycoconjugate vaccines containing chemically derived zwitterionic polysaccharides. AB - Group B Streptococcus (GBS) causes serious infection in neonates and is an important target of vaccine development. Zwitterionic polysaccharides (ZPS), obtained through chemical introduction of positive charges into anionic polysaccharides (PS) from GBS, have the ability to activate human and mouse antigen presenting cells (APCs) through toll-like receptor 2 (TLR2). To generate a polysaccharide vaccine with antigen (Ag) and adjuvant properties in one molecule, we have conjugated ZPS with a carrier protein. ZPS-glycoconjugates induce higher T-cell and Ab responses to carrier and PS, respectively, compared to control PS-glycoconjugates made with the native polysaccharide form. The increased immunogenicity of ZPS-conjugates correlates with their ability to activate dendritic cells (DCs). Moreover, protection of mothers or neonate offspring from lethal GBS challenge is better when mothers are immunized with ZPS conjugates compared to immunization with PS-conjugates. In TLR2 knockout mice, ZPS-conjugates lose both their increased immunogenicity and protective effect after vaccination. When ZPS are coadministered as adjuvants with unconjugated tetanus toxoid (TT), they have the ability to increase the TT-specific antibody titer. In conclusion, glycoconjugates containing ZPS are potent vaccines. They target Ag to TLR2-expressing APCs and activate these APCs, leading to better T cell priming and ultimately to higher protective Ab titers. Thus, rational chemical design can generate potent PS-adjuvants with wide application, including glycoconjugates and coadministration with unrelated protein Ags. PMID- 19805032 TI - Evidence for a ferryl intermediate in a heme-based dioxygenase. AB - In contrast to the wide spectrum of cytochrome P450 monooxygenases, there are only 2 heme-based dioxygenases in humans: tryptophan dioxygenase (hTDO) and indoleamine 2,3-dioxygenase (hIDO). hTDO and hIDO catalyze the same oxidative ring cleavage reaction of L-tryptophan to N-formyl kynurenine, the initial and rate-limiting step of the kynurenine pathway. Despite immense interest, the mechanism by which the 2 enzymes execute the dioxygenase reaction remains elusive. Here, we report experimental evidence for a key ferryl intermediate of hIDO that supports a mechanism in which the 2 atoms of dioxygen are inserted into the substrate via a consecutive 2-step reaction. This finding introduces a paradigm shift in our understanding of the heme-based dioxygenase chemistry, which was previously believed to proceed via simultaneous incorporation of both atoms of dioxygen into the substrate. The ferryl intermediate is not observable during the hTDO reaction, highlighting the structural differences between the 2 dioxygenases, as well as the importance of stereoelectronic factors in modulating the reactions. PMID- 19805033 TI - Evolution in health and medicine Sackler colloquium: Somatic evolutionary genomics: mutations during development cause highly variable genetic mosaicism with risk of cancer and neurodegeneration. AB - Somatic mutations must happen often during development because of the large number of cell divisions to expand from a single-cell zygote to a full organism. A mutation in development carries forward to all descendant cells, causing genetic mosaicism. Widespread genetic mosaicism may influence diseases that derive from a few genetically altered cells, such as cancer. I show how to predict the expected amount of mosaicism and the variation in mosaicism between individuals. I then calculate the predicted risk of cancer derived from developmental mutations. The calculations show that a significant fraction of cancer in later life likely arises from developmental mutations in early life. In addition, much of the variation in the risk of cancer between individuals may arise from variation in the degree of genetic mosaicism set in early life. I also suggest that certain types of neurodegeneration, such as amyotrophic lateral sclerosis (ALS), may derive from a small focus of genetically altered cells. If so, then the risk of ALS would be influenced by developmental mutations and the consequent variation in genetic mosaicism. New technologies promise the ability to measure genetic mosaicism by sampling a large number of cellular genomes within an individual. The sampling of many genomes within an individual will eventually allow one to reconstruct the cell lineage history of genetic change in a single body. Somatic evolutionary genomics will follow from this technology, providing new insight into the origin and progression of disease with increasing age. PMID- 19805034 TI - Crystal structure of native RPE65, the retinoid isomerase of the visual cycle. AB - Vertebrate vision is maintained by the retinoid (visual) cycle, a complex enzymatic pathway that operates in the retina to regenerate the visual chromophore, 11-cis-retinal. A key enzyme in this pathway is the microsomal membrane protein RPE65. This enzyme catalyzes the conversion of all-trans-retinyl esters to 11-cis-retinol in the retinal pigment epithelium (RPE). Mutations in RPE65 are known to be responsible for a subset of cases of the most common form of childhood blindness, Leber congenital amaurosis (LCA). Although retinoid isomerase activity has been attributed to RPE65, its catalytic mechanism remains a matter of debate. Also, the manner in which RPE65 binds to membranes and extracts retinoid substrates is unclear. To gain insight into these questions, we determined the crystal structure of native bovine RPE65 at 2.14-A resolution. The structural, biophysical, and biochemical data presented here provide the framework needed for an in-depth understanding of the mechanism of catalytic isomerization and membrane association, in addition to the role mutations that cause LCA have in disrupting protein function. PMID- 19805035 TI - A long-snouted, multihorned tyrannosaurid from the Late Cretaceous of Mongolia. AB - Tyrannosaurid theropods are characterized by a generalized body plan, and all well-known taxa possess deep and robust skulls that are optimized for exerting powerful bite forces. The fragmentary Late Cretaceous Alioramus appears to deviate from this trend, but its holotype and only known specimen is incomplete and poorly described. A remarkable new tyrannosaurid specimen from the Maastrichtian (Late Cretaceous) of Mongolia, including a nearly complete and well preserved skull and an extensive postcranium, represents a new species of Alioramus, Alioramus altai. This specimen conclusively demonstrates that Alioramus is a small, gracile, long-snouted carnivore that deviates from other tyrannosaurids in its body plan and presumably its ecological habits. As such, it increases the range of morphological diversity in one of the most familiar extinct clades. Phylogenetic analysis places Alioramus deep within the megapredatory Tyrannosauridae, and within the tyrannosaurine subclade that also includes Tarbosaurus and Tyrannosaurus. Both pneumatization and ornamentation are extreme compared with other tyrannosaurids, and the skull contains eight discrete horns. The new specimen is histologically aged at nine years old but is smaller than other tyrannosaurids of similar age. Despite its divergent cranial form, Alioramus is characterized by a similar sequence of ontogenetic changes as the megapredatory Tyrannosaurus and Albertosaurus, indicating that ontogenetic change is conservative in tyrannosaurids. PMID- 19805036 TI - Single-cell response to stiffness exhibits muscle-like behavior. AB - Living cells sense the rigidity of their environment and adapt their activity to it. In particular, cells cultured on elastic substrates align their shape and their traction forces along the direction of highest stiffness and preferably migrate towards stiffer regions. Although numerous studies investigated the role of adhesion complexes in rigidity sensing, less is known about the specific contribution of acto-myosin based contractility. Here we used a custom-made single-cell technique to measure the traction force as well as the speed of shortening of isolated myoblasts deflecting microplates of variable stiffness. The rate of force generation increased with increasing stiffness and followed a Hill force-velocity relationship. Hence, cell response to stiffness was similar to muscle adaptation to load, reflecting the force-dependent kinetics of myosin binding to actin. These results reveal an unexpected mechanism of rigidity sensing, whereby the contractile acto-myosin units themselves can act as sensors. This mechanism may translate anisotropy in substrate rigidity into anisotropy in cytoskeletal tension, and could thus coordinate local activity of adhesion complexes and guide cell migration along rigidity gradients. PMID- 19805037 TI - Birds track their Grinnellian niche through a century of climate change. AB - In the face of environmental change, species can evolve new physiological tolerances to cope with altered climatic conditions or move spatially to maintain existing physiological associations with particular climates that define each species' climatic niche. When environmental change occurs over short temporal and large spatial scales, vagile species are expected to move geographically by tracking their climatic niches through time. Here, we test for evidence of niche tracking in bird species of the Sierra Nevada mountains of California, focusing on 53 species resurveyed nearly a century apart at 82 sites on four elevational transects. Changes in climate and bird distributions resulted in focal species shifting their average climatological range over time. By comparing the directions of these shifts relative to the centroids of species' range-wide climatic niches, we found that 48 species (90.6%) tracked their climatic niche. Analysis of niche sensitivity on an independent set of occurrence data significantly predicted the temperature and precipitation gradients tracked by species. Furthermore, in 50 species (94.3%), site-specific occupancy models showed that the position of each site relative to the climatic niche centroid explained colonization and extinction probabilities better than a null model with constant probabilities. Combined, our results indicate that the factors limiting a bird species' range in the Sierra Nevada in the early 20th century also tended to drive changes in distribution over time, suggesting that climatic models derived from niche theory might be used successfully to forecast where and how to conserve species in the face of climate change. PMID- 19805038 TI - The transcription factors T-bet and GATA-3 control alternative pathways of T-cell differentiation through a shared set of target genes. AB - Upon detection of antigen, CD4(+) T helper (Th) cells can differentiate into a number of effector types that tailor the immune response to different pathogens. Alternative Th1 and Th2 cell fates are specified by the transcription factors T bet and GATA-3, respectively. Only a handful of target genes are known for these two factors and because of this, the mechanism through which T-bet and GATA-3 induce differentiation toward alternative cell fates is not fully understood. Here, we provide a genomic map of T-bet and GATA-3 binding in primary human T cells and identify their target genes, most of which are previously unknown. In Th1 cells, T-bet associates with genes of diverse function, including those with roles in transcriptional regulation, chemotaxis and adhesion. GATA-3 occupies genes in both Th1 and Th2 cells and, unexpectedly, shares a large proportion of targets with T-bet. Re-complementation of T-bet alters the expression of these genes in a manner that mirrors their differential expression between Th1 and Th2 lineages. These data show that the choice between Th1 and Th2 lineage commitment is the result of the opposing action of T-bet and GATA-3 at a shared set of target genes and may provide a general paradigm for the interaction of lineage specifying transcription factors. PMID- 19805039 TI - Infection of neurons and encephalitis after intracranial inoculation of herpes simplex virus requires the entry receptor nectin-1. AB - Multiple entry receptors can mediate infection of cells by herpes simplex virus (HSV), permitting alternative pathways for infection and disease. We investigated the roles of two known entry receptors, herpesvirus entry mediator (HVEM) and nectin-1, in infection of neurons in the CNS and the development of encephalitis. Wild-type, HVEM KO, nectin-1 KO, and HVEM/nectin-1 double KO mice were inoculated with HSV into the hippocampus. The mice were examined for development of encephalitis or were killed at various times after inoculation for immunohistological analyses of brain slices. Nectin-1 KO mice showed no signs of disease after intracranial inoculation, and no HSV antigens were detectable in the brain parenchyma. However, HSV antigens were detected in non-parenchymal cells lining the ventricles. In the double KO mice, there was also no disease and no detectable expression of viral antigens even in non-parenchymal cells, indicating that infection of these cells in the nectin-1 KO mice was dependent on the expression of HVEM. Wild-type and HVEM KO mice rapidly developed encephalitis, and the patterns of HSV replication in the brain were indistinguishable. Thus, expression of nectin-1 is necessary for HSV infection via the intracranial route and for encephalitis; HVEM is largely irrelevant. These results contrast with recent findings that (i) either HVEM or nectin-1 can permit HSV infection of the vaginal epithelium in mice and (ii) nectin-1 is not the sole receptor capable of enabling spread of HSV infection from the vaginal epithelium to the PNS and CNS. PMID- 19805040 TI - A superior descriptor of random textures and its predictive capacity. AB - Two-phase random textures abound in a host of contexts, including porous and composite media, ecological structures, biological media, and astrophysical structures. Questions surrounding the spatial structure of such textures continue to pose many theoretical challenges. For example, can two-point correlation functions be identified that can be manageably measured and yet reflect nontrivial higher-order structural information about the textures? We present a solution to this question by probing the information content of the widest class of different types of two-point functions examined to date using inverse "reconstruction" techniques. This enables us to show that a superior descriptor is the two-point cluster function C(2)(r), which is sensitive to topological connectedness information. We demonstrate the utility of C(2)(r) by accurately reconstructing textures drawn from materials science, cosmology, and granular media, among other examples. Our work suggests a theoretical pathway to predict the bulk physical properties of random textures and that also has important ramifications for atomic and molecular systems. PMID- 19805041 TI - Niches and distributional areas: concepts, methods, and assumptions. AB - Estimating actual and potential areas of distribution of species via ecological niche modeling has become a very active field of research, yet important conceptual issues in this field remain confused. We argue that conceptual clarity is enhanced by adopting restricted definitions of "niche" that enable operational definitions of basic concepts like fundamental, potential, and realized niches and potential and actual distributional areas. We apply these definitions to the question of niche conservatism, addressing what it is that is conserved and showing with a quantitative example how niche change can be measured. In this example, we display the extremely irregular structure of niche space, arguing that it is an important factor in understanding niche evolution. Many cases of apparently successful models of distributions ignore biotic factors: we suggest explanations to account for this paradox. Finally, relating the probability of observing a species to ecological factors, we address the issue of what objects are actually calculated by different niche modeling algorithms and stress the fact that methods that use only presence data calculate very different quantities than methods that use absence data. We conclude that the results of niche modeling exercises can be interpreted much better if the ecological and mathematical assumptions of the modeling process are made explicit. PMID- 19805042 TI - Phylogenetic structure in tropical hummingbird communities. AB - How biotic interactions, current and historical environment, and biogeographic barriers determine community structure is a fundamental question in ecology and evolution, especially in diverse tropical regions. To evaluate patterns of local and regional diversity, we quantified the phylogenetic composition of 189 hummingbird communities in Ecuador. We assessed how species and phylogenetic composition changed along environmental gradients and across biogeographic barriers. We show that humid, low-elevation communities are phylogenetically overdispersed (coexistence of distant relatives), a pattern that is consistent with the idea that competition influences the local composition of hummingbirds. At higher elevations communities are phylogenetically clustered (coexistence of close relatives), consistent with the expectation of environmental filtering, which may result from the challenge of sustaining an expensive means of locomotion at high elevations. We found that communities in the lowlands on opposite sides of the Andes tend to be phylogenetically similar despite their large differences in species composition, a pattern implicating the Andes as an important dispersal barrier. In contrast, along the steep environmental gradient between the lowlands and the Andes we found evidence that species turnover is comprised of relatively distantly related species. The integration of local and regional patterns of diversity across environmental gradients and biogeographic barriers provides insight into the potential underlying mechanisms that have shaped community composition and phylogenetic diversity in one of the most species-rich, complex regions of the world. PMID- 19805043 TI - Coin hoards speak of population declines in Ancient Rome. AB - In times of violence, people tend to hide their valuables, which are later recovered unless the owners had been killed or driven away. Thus, the temporal distribution of unrecovered coin hoards is an excellent proxy for the intensity of internal warfare. We use this relationship to resolve a long-standing controversy in Roman history. Depending on who was counted in the early Imperial censuses (adult males or the entire citizenry including women and minors), the Roman citizen population of Italy either declined, or more than doubled, during the first century BCE. This period was characterized by a series of civil wars, and historical evidence indicates that high levels of sociopolitical instability are associated with demographic contractions. We fitted a simple model quantifying the effect of instability (proxied by hoard frequency) on population dynamics to the data before 100 BCE. The model predicts declining population after 100 BCE. This suggests that the vigorous growth scenario is highly implausible. PMID- 19805044 TI - Unseen facial and bodily expressions trigger fast emotional reactions. AB - The spontaneous tendency to synchronize our facial expressions with those of others is often termed emotional contagion. It is unclear, however, whether emotional contagion depends on visual awareness of the eliciting stimulus and which processes underlie the unfolding of expressive reactions in the observer. It has been suggested either that emotional contagion is driven by motor imitation (i.e., mimicry), or that it is one observable aspect of the emotional state arising when we see the corresponding emotion in others. Emotional contagion reactions to different classes of consciously seen and "unseen" stimuli were compared by presenting pictures of facial or bodily expressions either to the intact or blind visual field of two patients with unilateral destruction of the visual cortex and ensuing phenomenal blindness. Facial reactions were recorded using electromyography, and arousal responses were measured with pupil dilatation. Passive exposure to unseen expressions evoked faster facial reactions and higher arousal compared with seen stimuli, therefore indicating that emotional contagion occurs also when the triggering stimulus cannot be consciously perceived because of cortical blindness. Furthermore, stimuli that are very different in their visual characteristics, such as facial and bodily gestures, induced highly similar expressive responses. This shows that the patients did not simply imitate the motor pattern observed in the stimuli, but resonated to their affective meaning. Emotional contagion thus represents an instance of truly affective reactions that may be mediated by visual pathways of old evolutionary origin bypassing cortical vision while still providing a cornerstone for emotion communication and affect sharing. PMID- 19805045 TI - Regulation of ciliary polarity by the APC/C. AB - Planar cell polarity signaling controls a variety of polarized cell behaviors. In multiciliated Xenopus epidermal cells, recruitment of Dishevelled (Dvl) to the basal body and its localization to the center of the ciliary rootlet are required to correctly position the motile cilia. We now report that the anaphase-promoting complex (APC/C) recognizes a D-box motif of Dvl and ubiquitylates Dvl on a highly conserved lysine residue. Inhibition of APC/C function by knockdown of the ANAPC2 subunit disrupts the polarity of motile cilia and alters the directionality of the fluid movement along the epidermis of the Xenopus embryo. Our results suggest that the APC/C activity enables cilia to correctly polarize in Xenopus epidermal cells. PMID- 19805046 TI - A little bit of lithium does a lot for hydrogen. AB - From detailed assessments of electronic structure, we find that a combination of significantly quantal elements, six of seven atoms being hydrogen, becomes a stable metal at a pressure approximately 1/4 of that required to metalize pure hydrogen itself. The system, LiH(6) (and other LiH(n)), may well have extensions beyond the constituent lithium. These hypothetical materials demonstrate that nontraditional stoichiometries can considerably expand the view of chemical combination under moderate pressure. PMID- 19805047 TI - Possible freshwater resource consumption by the earliest directly dated European modern humans: Implications for direct radiometric dating. PMID- 19805048 TI - miR-145 directs intestinal maturation in zebrafish. AB - The rapid specification and differentiation of the embryonic zebrafish gut is essential to provide contractility for the digestion of food. The role of microRNAs in modulating gut epithelial or smooth muscle differentiation is currently not known. Here we show that the microRNA miR-145 is strongly expressed in zebrafish gut smooth muscle and regulates its development. Modulation of miR 145 levels results in gut smooth muscle and epithelium maturation defects. Loss of miR-145 results in defects of smooth muscle function as measured by decreased nitric oxide production but also leads to increased expression of the embryonic smooth muscle markers sm22alpha-b, nm-mhc-b, and smoothelin. Defects in gut epithelial maturation are also present as observed by immature morphology and a complete loss of alkaline phosphatase expression. Loss or gain of miR-145 function phenocopies defects observed with altered gata6 expression and accordingly, we show that miR-145 directly represses gata6, and that gata6 is a major miR-145 target in vitro and in vivo. miR-145 therefore plays a critical role in promoting the maturation of both layers of the gut during development through regulation of gata6. PMID- 19805049 TI - Flow and diffusion of high-stakes test scores. AB - We apply visualization and modeling methods for convective and diffusive flows to public school mathematics test scores from Texas. We obtain plots that show the most likely future and past scores of students, the effects of random processes such as guessing, and the rate at which students appear in and disappear from schools. We show that student outcomes depend strongly upon economic class, and identify the grade levels where flows of different groups diverge most strongly. Changing the effectiveness of instruction in one grade naturally leads to strongly nonlinear effects on student outcomes in subsequent grades. PMID- 19805050 TI - Decoding cognitive control in human parietal cortex. AB - Efficient execution of perceptual-motor tasks requires rapid voluntary reconfiguration of cognitive task sets as circumstances unfold. Such acts of cognitive control, which are thought to rely on a network of cortical regions in prefrontal and posterior parietal cortex, include voluntary shifts of attention among perceptual inputs or among memory representations, or switches between categorization or stimulus-response mapping rules. A critical unanswered question is whether task set shifts in these different domains are controlled by a common, domain-independent mechanism or by separate, domain-specific mechanisms. Recent studies have implicated a common region of medial superior parietal lobule (mSPL) as a domain-independent source of cognitive control during shifts between perceptual, mnemonic, and rule representations. Here, we use fMRI and event related multivoxel pattern classification to show that spatial patterns of brain activity within mSPL reliably express which of several domains of cognitive control is at play on a moment-by-moment basis. Critically, these spatiotemporal brain patterns are stable over time within subjects tested several months apart and across a variety of tasks, including shifting visuospatial attention, switching categorization rules, and shifting attention in working memory. PMID- 19805051 TI - Identifying genotype-dependent efficacy of single and combined PI3K- and MAPK pathway inhibition in cancer. AB - In cancer, genetically activated proto-oncogenes often induce "upstream" dependency on the activity of the mutant oncoprotein. Therapeutic inhibition of these activated oncoproteins can induce massive apoptosis of tumor cells, leading to sometimes dramatic tumor regressions in patients. The PI3K and MAPK signaling pathways are central regulators of oncogenic transformation and tumor maintenance. We hypothesized that upstream dependency engages either one of these pathways preferentially to induce "downstream" dependency. Therefore, we analyzed whether downstream pathway dependency segregates by genetic aberrations upstream in lung cancer cell lines. Here, we show by systematically linking drug response to genomic aberrations in non-small-cell lung cancer, as well as in cell lines of other tumor types and in a series of in vivo cancer models, that tumors with genetically activated receptor tyrosine kinases depend on PI3K signaling, whereas tumors with mutations in the RAS/RAF axis depend on MAPK signaling. However, efficacy of downstream pathway inhibition was limited by release of negative feedback loops on the reciprocal pathway. By contrast, combined blockade of both pathways was able to overcome the reciprocal pathway activation induced by inhibitor-mediated release of negative feedback loops and resulted in a significant increase in apoptosis and tumor shrinkage. Thus, by using a systematic chemo-genomics approach, we identify genetic lesions connected to PI3K and MAPK pathway activation and provide a rationale for combined inhibition of both pathways. Our findings may have implications for patient stratification in clinical trials. PMID- 19805052 TI - Evolution in health and medicine Sackler colloquium: Consanguinity, human evolution, and complex diseases. AB - There is little information on inbreeding during the critical early years of human existence. However, given the small founding group sizes and restricted mate choices it seems inevitable that intrafamilial reproduction occurred and the resultant levels of inbreeding would have been substantial. Currently, couples related as second cousins or closer (F >or= 0.0156) and their progeny account for an estimated 10.4% of the global population. The highest rates of consanguineous marriage occur in north and sub-Saharan Africa, the Middle East, and west, central, and south Asia. In these regions even couples who regard themselves as unrelated may exhibit high levels of homozygosity, because marriage within clan, tribe, caste, or biraderi boundaries has been a long-established tradition. Mortality in first-cousin progeny is approximately 3.5% higher than in nonconsanguineous offspring, although demographic, social, and economic factors can significantly influence the outcome. Improving socioeconomic conditions and better access to health care will impact the effects of consanguinity, with a shift from infant and childhood mortality to extended morbidity. At the same time, a range of primarily social factors, including urbanization, improved female education, and smaller family sizes indicate that the global prevalence of consanguineous unions will decline. This shift in marriage patterns will initially result in decreased homozygosity, accompanied by a reduction in the expression of recessive single-gene disorders. Although the roles of common and rare gene variants in the etiology of complex disease remain contentious, it would be expected that declining consanguinity would also be reflected in reduced prevalence of complex diseases, especially in population isolates. PMID- 19805053 TI - How to make a meandering river. PMID- 19805054 TI - Genetic engineering of human stem cells for enhanced angiogenesis using biodegradable polymeric nanoparticles. AB - Stem cells hold great potential as cell-based therapies to promote vascularization and tissue regeneration. However, the use of stem cells alone to promote angiogenesis remains limited because of insufficient expression of angiogenic factors and low cell viability after transplantation. Here, we have developed vascular endothelial growth factor (VEGF) high-expressing, transiently modified stem cells for the purposes of promoting angiogenesis. Nonviral, biodegradable polymeric nanoparticles were developed to deliver hVEGF gene to human mesenchymal stem cells (hMSCs) and human embryonic stem cell-derived cells (hESdCs). Treated stem cells demonstrated markedly enhanced hVEGF production, cell viability, and engraftment into target tissues. S.c. implantation of scaffolds seeded with VEGF-expressing stem cells (hMSCs and hESdCs) led to 2- to 4-fold-higher vessel densities 2 weeks after implantation, compared with control cells or cells transfected with VEGF by using Lipofectamine 2000, a leading commercial reagent. Four weeks after intramuscular injection into mouse ischemic hindlimbs, genetically modified hMSCs substantially enhanced angiogenesis and limb salvage while reducing muscle degeneration and tissue fibrosis. These results indicate that stem cells engineered with biodegradable polymer nanoparticles may be therapeutic tools for vascularizing tissue constructs and treating ischemic disease. PMID- 19805055 TI - Assembly mechanisms of RNA pseudoknots are determined by the stabilities of constituent secondary structures. AB - Understanding how RNA molecules navigate their rugged folding landscapes holds the key to describing their roles in a variety of cellular functions. To dissect RNA folding at the molecular level, we performed simulations of three pseudoknots (MMTV and SRV-1 from viral genomes and the hTR pseudoknot from human telomerase) using coarse-grained models. The melting temperatures from the specific heat profiles are in good agreement with the available experimental data for MMTV and hTR. The equilibrium free energy profiles, which predict the structural transitions that occur at each melting temperature, are used to propose that the relative stabilities of the isolated helices control their folding mechanisms. Kinetic simulations, which corroborate the inferences drawn from the free energy profiles, show that MMTV folds by a hierarchical mechanism with parallel paths, i.e., formation of one of the helices nucleates the assembly of the rest of the structure. The SRV-1 pseudoknot, which folds in a highly cooperative manner, assembles in a single step in which the preformed helices coalesce nearly simultaneously to form the tertiary structure. Folding occurs by multiple pathways in the hTR pseudoknot, the isolated structural elements of which have similar stabilities. In one of the paths, tertiary interactions are established before the formation of the secondary structures. Our work shows that there are significant sequence-dependent variations in the folding landscapes of RNA molecules with similar fold. We also establish that assembly mechanisms can be predicted using the stabilities of the isolated secondary structures. PMID- 19805056 TI - Small RNAs serve as a genetic buffer against genomic shock in Arabidopsis interspecific hybrids and allopolyploids. AB - Small RNAs, including microRNAs (miRNAs), small interfering RNAs (siRNAs), and trans-acting siRNAs (tasiRNAs), control gene expression and epigenetic regulation. Although the roles of miRNAs and siRNAs have been extensively studied, their expression diversity and evolution in closely related species and interspecific hybrids are poorly understood. Here, we show comprehensive analyses of miRNA expression and siRNA distributions in two closely related species Arabidopsis thaliana and Arabidopsis arenosa, a natural allotetraploid Arabidopsis suecica, and two resynthesized allotetraploid lines (F(1) and F(7)) derived from A. thaliana and A. arenosa. We found that repeat- and transposon associated siRNAs were highly divergent between A. thaliana and A. arenosa. A. thaliana siRNA populations underwent rapid changes in F(1) but were stably maintained in F(7) and A. suecica. The correlation between siRNAs and nonadditive gene expression in allopolyploids is insignificant. In contrast, miRNA and tasiRNA sequences were conserved between species, but their expression patterns were highly variable between the allotetraploids and their progenitors. Many miRNAs tested were nonadditively expressed (deviating from the mid-parent value, MPV) in the allotetraploids and triggered unequal degradation of A. thaliana or A. arenosa targets. The data suggest that small RNAs produced during interspecific hybridization or polyploidization serve as a buffer against the genomic shock in interspecific hybrids and allopolyploids: Stable inheritance of repeat-associated siRNAs maintains chromatin and genome stability, whereas expression variation of miRNAs leads to changes in gene expression, growth vigor, and adaptation. PMID- 19805057 TI - Modeling children's early grammatical knowledge. AB - Theories of grammatical development differ in how much abstract knowledge they attribute to young children. Here, we report a series of experiments using a computational model to evaluate the explanatory power of child grammars based not on abstract rules but on concrete words and phrases and some local abstractions associated with these words and phrases. We use a Bayesian procedure to extract such item-based grammars from transcriptions of 28+ h of each of two children's speech at 2 and 3 years of age. We then use these grammars to parse all of the unique multiword utterances from transcriptions of separate recordings of these same children at each of the two ages. We found that at 2 years of age such a model had good coverage and predictive fit, with the children showing radically limited productivity. Furthermore, adding expert-annotated parts of speech to the induction procedure had little effect on coverage, with the exception of the category of noun. At age 3, the children's productivity sharply increased and the addition of a verb and a noun category markedly improved the model's performance. PMID- 19805058 TI - Nonrelatives inherit colony resources in a primitive termite. AB - The evolution of eusociality, especially how selection would favor sterility or subfertility of most individuals within a highly social colony, is an unresolved paradox. Eusociality evolved independently in diverse taxa, including insects (all ants and termites; some bees, wasps, thrips, and beetles), snapping shrimp, and naked mole rats. Termites have received comparatively less focus than the haplodiploid Hymenoptera (ants, bees, and wasps); however, they are the only diploid group with highly complex colonies and an extraordinary diversity of castes. In this study we staged encounters between unrelated colonies of primitive dampwood termites, Zootermopsis nevadensis, mimicking natural meetings that occur under bark. During encounters, kings and/or queens were killed and surviving members merged into one colony. After encounters, members of both unrelated colonies cooperated as a single social unit. We determined the colony of origin of replacement reproductives that emerged after death of kings and/or queens. Here, we document that replacement reproductives developed from workers in either or both original colonies, inherited the merged resources of the colony, and sometimes interbred. Because this species shares many characteristics with ancestral termites, these findings demonstrate how ecological factors could have promoted the evolution of eusociality by accelerating and enhancing direct fitness opportunities of helper offspring, rendering relatedness favoring kin selection less critical. PMID- 19805059 TI - Glucocorticoid regulation of the circadian clock modulates glucose homeostasis. AB - Circadian clock genes are regulated by glucocorticoids; however, whether this regulation is a direct or secondary effect and the physiological consequences of this regulation were unknown. Here, we identified glucocorticoid response elements (GREs) at multiple clock genes and showed that 3 were directly regulated by the glucocorticoid receptor. We determined that a GRE within the core clock gene Per2 was continuously occupied during rhythmic expression and essential for glucocorticoid regulation of that gene in vivo. We further demonstrated that mice with a genomic deletion spanning this GRE expressed elevated leptin levels and were protected from glucose intolerance and insulin resistance on glucocorticoid treatment but not from muscle wasting. We conclude that Per2 is an integral component of a particular glucocorticoid regulatory pathway and that glucocorticoid regulation of the peripheral clock is selectively required for some actions of glucocorticoids. PMID- 19805060 TI - Transient dwarfism of soil fauna during the Paleocene-Eocene Thermal Maximum. AB - Soil organisms, as recorded by trace fossils in paleosols of the Willwood Formation, Wyoming, show significant body-size reductions and increased abundances during the Paleocene-Eocene Thermal Maximum (PETM). Paleobotanical, paleopedologic, and oxygen isotope studies indicate high temperatures during the PETM and sharp declines in precipitation compared with late Paleocene estimates. Insect and oligochaete burrows increase in abundance during the PETM, suggesting longer periods of soil development and improved drainage conditions. Crayfish burrows and molluscan body fossils, abundant below and above the PETM interval, are significantly less abundant during the PETM, likely because of drier floodplain conditions and lower water tables. Burrow diameters of the most abundant ichnofossils are 30-46% smaller within the PETM interval. As burrow size is a proxy for body size, significant reductions in burrow diameter suggest that their tracemakers were smaller bodied. Smaller body sizes may have resulted from higher subsurface temperatures, lower soil moisture conditions, or nutritionally deficient vegetation in the high-CO(2) atmosphere inferred for the PETM. Smaller soil fauna co-occur with dwarf mammal taxa during the PETM; thus, a common forcing mechanism may have selected for small size in both above- and below ground terrestrial communities. We predict that soil fauna have already shown reductions in size over the last 150 years of increased atmospheric CO(2) and surface temperatures or that they will exhibit this pattern over the next century. We retrodict also that soil fauna across the Permian-Triassic and Triassic-Jurassic boundary events show significant size decreases because of similar forcing mechanisms driven by rapid global warming. PMID- 19805061 TI - Learning sculpts the spontaneous activity of the resting human brain. AB - The brain is not a passive sensory-motor analyzer driven by environmental stimuli, but actively maintains ongoing representations that may be involved in the coding of expected sensory stimuli, prospective motor responses, and prior experience. Spontaneous cortical activity has been proposed to play an important part in maintaining these ongoing, internal representations, although its functional role is not well understood. One spontaneous signal being intensely investigated in the human brain is the interregional temporal correlation of the blood-oxygen level-dependent (BOLD) signal recorded at rest by functional MRI (functional connectivity-by-MRI, fcMRI, or BOLD connectivity). This signal is intrinsic and coherent within a number of distributed networks whose topography closely resembles that of functional networks recruited during tasks. While it is apparent that fcMRI networks reflect anatomical connectivity, it is less clear whether they have any dynamic functional importance. Here, we demonstrate that visual perceptual learning, an example of adult neural plasticity, modifies the resting covariance structure of spontaneous activity between networks engaged by the task. Specifically, after intense training on a shape-identification task constrained to one visual quadrant, resting BOLD functional connectivity and directed mutual interaction between trained visual cortex and frontal-parietal areas involved in the control of spatial attention were significantly modified. Critically, these changes correlated with the degree of perceptual learning. We conclude that functional connectivity serves a dynamic role in brain function, supporting the consolidation of previous experience. PMID- 19805062 TI - Folding of electrostatically charged beads-on-a-string as an experimental realization of a theoretical model in polymer science. AB - The "beads-on-a-string" model for folding of polymers is a cornerstone of theoretical polymer science. This communication describes a physical model of beads-on-a-string, based on the folding of flexible strings of electrostatically charged beads in two dimensions. The system comprises millimeter-scale Teflon and Nylon-6,6 (spherical or cylindrical) beads (approximately 6 mm in diameter) separated by smaller (approximately 3 mm) poly(methyl methacrylate) (PMMA) spherical beads, threaded on a flexible string. The smaller, uncharged beads define the distances between the larger beads, and control the flexibility of the string. During agitation of the sequence of beads on a planar, horizontal paper surface, tribocharging generates opposite electrostatic charges on the larger Nylon and Teflon beads, but leaves the smaller PMMA beads essentially uncharged; the resulting electrostatic interactions cause the string to fold. Examination and comparison of two models--one physical and one theoretical--may offer a new approach to understanding folding, collapse, and molecular recognition at an abstract level, with particular opportunity to explore the influence of the flexibility of the string and the shape of the beads on the pattern and rate of folding. The physical system is, thus, an analog computer, simulating the theoretical beads-on-a-string model in two dimensions; this system makes it possible to test hypotheses connecting "sequence" to "folding", rapidly and conveniently, while exploring nonlinearities and other complexities omitted from the theoretical model. PMID- 19805063 TI - Voltage-gated proton channels maintain pH in human neutrophils during phagocytosis. AB - Phagocytosis of microbial invaders represents a fundamental defense mechanism of the innate immune system. The subsequent killing of microbes is initiated by the respiratory burst, in which nicotinamide adenine dinucleotide phosphate (NADPH) oxidase generates vast amounts of superoxide anion, precursor to bactericidal reactive oxygen species. Cytoplasmic pH regulation is crucial because NADPH oxidase functions optimally at neutral pH, yet produces enormous quantities of protons. We monitored pH(i) in individual human neutrophils during phagocytosis of opsonized zymosan, using confocal imaging of the pH sensing dye SNARF-1, enhanced by shifted excitation and emission ratioing, or SEER. Despite long standing dogma that Na(+)/H(+) antiport regulates pH during the phagocyte respiratory burst, we show here that voltage-gated proton channels are the first transporter to respond. During the initial phagocytotic event, pH(i) decreased sharply, and recovery required both Na(+)/H(+) antiport and proton current. Inhibiting myeloperoxidase attenuated the acidification, suggesting that diffusion of HOCl into the cytosol comprises a substantial acid load. Inhibiting proton channels with Zn(2+) resulted in profound acidification to levels that inhibit NADPH oxidase. The pH changes accompanying phagocytosis in bone marrow phagocytes from HVCN1-deficient mice mirrored those in control mouse cells treated with Zn(2+). Both the rate and extent of acidification in HVCN1-deficient cells were twice larger than in control cells. In summary, acid extrusion by proton channels is essential to the production of reactive oxygen species during phagocytosis. PMID- 19805064 TI - Spectroelectrochemical determination of the redox potential of pheophytin a, the primary electron acceptor in photosystem II. AB - Thin-layer cell spectroelectrochemistry, featuring rigorous potential control and rapid redox equilibration within the cell, was used to measure the redox potential E(m)(Phe a/Phe a(-)) of pheophytin (Phe) a, the primary electron acceptor in an oxygen-evolving photosystem (PS) II core complex from a thermophilic cyanobacterium Thermosynechococcus elongatus. Interferences from dissolved O(2) and water reductions were minimized by airtight sealing of the sample cell added with dithionite and mercury plating on the gold minigrid working electrode surface, respectively. The result obtained at a physiological pH of 6.5 was E(m)(Phe a/Phe a(-)) = -505 + or - 6 mV vs. SHE, which is by approximately 100 mV more positive than the values measured approximately 30 years ago at nonphysiological pH and widely accepted thereafter in the field of photosynthesis research. Using the P680* - Phe a free energy difference, as estimated from kinetic analyses by previous authors, the present result would locate the E(m)(P680/P680(+)) value, which is one of the key parameters but still resists direct measurements, at approximately +1,210 mV. In view of these pieces of information, a renewed diagram is proposed for the energetics in PS II. PMID- 19805065 TI - Cargo binding induces dimerization of myosin VI. AB - Although myosin VI has properties that would allow it to function optimally as a dimer, full-length myosin VI exists as a monomer in isolation. Based on the ability of myosin VI monomers to dimerize when held in close proximity, we postulated that cargo binding normally regulates dimerization of myosin VI. We tested this hypothesis by expressing a known dimeric cargo adaptor protein of myosin VI, optineurin, and the myosin VI-binding segment from a monomeric cargo adaptor protein, Dab2. In the presence of these adaptor proteins, full-length myosin VI has ATPase properties of a dimer, appears as a dimer in electron micrographs, and moves processively on actin filaments. The results support a model in which cargo binding exposes internal dimerization sequences within full length myosin VI. Because, unexpectedly, a monomeric fragment of Dab2 triggers dimerization, it would appear that myosin VI is designed to function as a dimer in cells. PMID- 19805066 TI - Evolutionary replacement of UV vision by violet vision in fish. AB - The vertebrate ancestor possessed ultraviolet (UV) vision and many species have retained it during evolution. Many other species switched to violet vision and, then again, some avian species switched back to UV vision. These UV and violet vision are mediated by short wavelength-sensitive (SWS1) pigments that absorb light maximally (lambda(max)) at approximately 360 and 390-440 nm, respectively. It is not well understood why and how these functional changes have occurred. Here, we cloned the pigment of scabbardfish (Lepidopus fitchi) with a lambda(max) of 423 nm, an example of violet-sensitive SWS1 pigment in fish. Mutagenesis experiments and quantum mechanical/molecular mechanical (QM/MM) computations show that the violet-sensitivity was achieved by the deletion of Phe-86 that converted the unprotonated Schiff base-linked 11-cis-retinal to a protonated form. The finding of a violet-sensitive SWS1 pigment in scabbardfish suggests that many other fish also have orthologous violet pigments. The isolation and comparison of such violet and UV pigments in fish living in different ecological habitats will open an unprecedented opportunity to elucidate not only the molecular basis of phenotypic adaptations, but also the genetics of UV and violet vision. PMID- 19805067 TI - Differential utilization of T cell receptor TCR alpha/TCR delta locus variable region gene segments is mediated by accessibility. AB - T cell receptor (TCR) variable region exons are assembled from germline V, (D), and J gene segments, each of which is flanked by recombination signal (RS) sequences that are composed of a conserved heptamer, a spacer of 12 or 23 bp, and a characteristic nonamer. V(D)J recombination only occurs between V, D, and J segments flanked by RS sequences that contain, respectively, 12(12-RS)- and 23(23 RS)-bp spacers (12/23 rule). Additional mechanisms can restrict joining of 12/23 RS matched segments beyond the 12/23 rule (B12/23). The TCRdelta locus is contained within the TCRalpha locus; TCRalpha variable region exons are encoded by TRAV and TRAJ segments and those of TCRdelta by TRDV, TRDD, and TRDJ segments. On the basis of the 12/23 rule, both TRAV and TRDV gene segments are compatible to rearrange with TRDD gene segments; however, TRAV-to-TRDD joins are not observed in vivo. Absence of TRAV-to-TRDD rearrangement might be explained either by B12/23 restriction or by differential accessibility of the TRDV versus TRAV gene segments for rearrangement to TRDD. We used in vitro substrate analysis to reveal that both TRAV and TRDV 23-RSs mediate rearrangements to the 5'TRDD1 12 RS, demonstrating that B12/23 restriction does not explain these rearrangement biases. However, targeted replacement of TRDD1 and its 12-RSs with TRAJ38 and its 12-RS showed that TRDV gene segments rearrange with the ectopic TRAJ38, whereas TRAV segments do not. Our results demonstrate that sorting of TRAV and TRDV gene segments is determined by differential locus accessibility during T cell development. PMID- 19805068 TI - How oxygen attacks [FeFe] hydrogenases from photosynthetic organisms. AB - Green algae such as Chlamydomonas reinhardtii synthesize an [FeFe] hydrogenase that is highly active in hydrogen evolution. However, the extreme sensitivity of [FeFe] hydrogenases to oxygen presents a major challenge for exploiting these organisms to achieve sustainable photosynthetic hydrogen production. In this study, the mechanism of oxygen inactivation of the [FeFe] hydrogenase CrHydA1 from C. reinhardtii has been investigated. X-ray absorption spectroscopy shows that reaction with oxygen results in destruction of the [4Fe-4S] domain of the active site H-cluster while leaving the di-iron domain (2Fe(H)) essentially intact. By protein film electrochemistry we were able to determine the order of events leading up to this destruction. Carbon monoxide, a competitive inhibitor of CrHydA1 which binds to an Fe atom of the 2Fe(H) domain and is otherwise not known to attack FeS clusters in proteins, reacts nearly two orders of magnitude faster than oxygen and protects the enzyme against oxygen damage. These results therefore show that destruction of the [4Fe-4S] cluster is initiated by binding and reduction of oxygen at the di-iron domain-a key step that is blocked by carbon monoxide. The relatively slow attack by oxygen compared to carbon monoxide suggests that a very high level of discrimination can be achieved by subtle factors such as electronic effects (specific orbital overlap requirements) and steric constraints at the active site. PMID- 19805069 TI - Cell surface-bound IL-1alpha is an upstream regulator of the senescence associated IL-6/IL-8 cytokine network. AB - Inflammation underlies most age-related diseases, including cancer, but the etiology is poorly understood. One proposed factor is the presence of senescent cells, which increase with age. The senescence response arrests the proliferation of potentially oncogenic cells, and most senescent cells secrete high levels of proinflammatory cytokines and other proteins. The complex senescence-associated secretory phenotype is likely regulated at multiple levels, most of which are unknown. We show that cell surface-bound IL-1alpha is essential for signaling the senescence-associated secretion of IL-6 and IL-8, 2 proinflammatory cytokines that also reinforce the senescence growth arrest. Senescent human fibroblasts expressed high levels of IL-1alpha mRNA, intracellular protein, and cell surface associated protein, but secreted very little protein. An IL-1 receptor (IL1R) antagonist, neutralizing IL-1alpha antibodies, and IL-1alpha depletion by RNA interference all markedly reduced senescence-associated IL-6/IL-8 secretion. Depletion of the key IL-1R signaling component IRAK1 also suppressed this secretion, and IL-1alpha neutralizing antibodies prevented IRAK1 degradation, indicating engagement of the IL-1R signaling pathway. Furthermore, IL-1alpha depletion reduced the DNA binding activity of NF-kappaB and C/EBPbeta, which stimulate IL-6/IL-8 transcription. IL-1alpha was a general regulator of senescence-associated IL-6/IL-8 secretion because IL-1alpha blockade reduced IL 6/IL-8 secretion whether cells senesced owing to DNA damage, replicative exhaustion, oncogenic RAS, or chromatin relaxation. Furthermore, conditioned medium from IL-1alpha-depleted senescent cells markedly reduced the IL-6/IL-8 dependent invasiveness of metastatic cancer cells, indicating that IL-1alpha regulates the biological effects of these cytokines. Thus, cell surface IL-1alpha is an essential cell-autonomous regulator of the senescence-associated IL-6/IL-8 cytokine network. PMID- 19805070 TI - Natural and synthetic prion structure from X-ray fiber diffraction. AB - A conformational isoform of the mammalian prion protein (PrP(Sc)) is the sole component of the infectious pathogen that causes the prion diseases. We have obtained X-ray fiber diffraction patterns from infectious prions that show cross beta diffraction: meridional intensity at 4.8 A resolution, indicating the presence of beta strands running approximately at right angles to the filament axis and characteristic of amyloid structure. Some of the patterns also indicated the presence of a repeating unit along the fiber axis, corresponding to four beta strands. We found that recombinant (rec) PrP amyloid differs substantially from highly infectious brain-derived prions, both in structure as demonstrated by the diffraction data, and in heterogeneity as shown by electron microscopy. In addition to the strong 4.8 A meridional reflection, the recPrP amyloid diffraction is characterized by strong equatorial intensity at approximately 10.5 A, absent from brain-derived prions, and indicating the presence of stacked beta sheets. Synthetic prions recovered from transgenic mice inoculated with recPrP amyloid displayed structural characteristics and homogeneity similar to those of naturally occurring prions. The relationship between the structural differences and prion infectivity is uncertain, but might be explained by any of several hypotheses: only a minority of recPrP amyloid possesses a replication-competent conformation, the majority of recPrP amyloid has to undergo a conformational maturation to acquire replication competency, or inhibitory forms of recPrP amyloid interfere with replication during the initial transmission. PMID- 19805071 TI - Groucho corepressor functions as a cofactor for the Knirps short-range transcriptional repressor. AB - Despite the pervasive roles for repressors in transcriptional control, the range of action of these proteins on cis regulatory elements remains poorly understood. Knirps has essential roles in patterning the Drosophila embryo by means of short range repression, an activity that is essential for proper regulation of complex transcriptional control elements. Short-range repressors function in a local fashion to interfere with the activity of activators or basal promoters within approximately 100 bp. In contrast, long-range repressors such as Hairy act over distances >1 kb. The functional distinction between these two classes of repressors has been suggested to stem from the differential recruitment of the CtBP corepressor to short-range repressors and Groucho to long-range repressors. Contrary to this differential recruitment model, we report that Groucho is a functional part of the Knirps short-range repression complex. The corepressor interaction is mediated via an eh-1 like motif present in the N terminus and a conserved region present in the central portion of Knirps. We also show that this interaction is important for the CtBP-independent repression activity of Knirps and is required for regulation of even-skipped. Our study uncovers a previously uncharacterized interaction between proteins previously thought to function in distinct repression pathways, and indicates that the Groucho corepressor can be differentially harnessed to execute short- and long-range repression. PMID- 19805072 TI - Crystal structure of Manduca sexta prophenoloxidase provides insights into the mechanism of type 3 copper enzymes. AB - Arthropod phenoloxidase (PO) generates quinones and other toxic compounds to sequester and kill pathogens during innate immune responses. It is also involved in wound healing and other physiological processes. Insect PO is activated from its inactive precursor, prophenoloxidase (PPO), by specific proteolysis via a serine protease cascade. Here, we report the crystal structure of PPO from a lepidopteran insect at a resolution of 1.97 A, which is the initial structure for a PPO from the type 3 copper protein family. Manduca sexta PPO is a heterodimer consisting of 2 homologous polypeptide chains, PPO1 and PPO2. The active site of each subunit contains a canonical type 3 di-nuclear copper center, with each copper ion coordinated with 3 structurally conserved histidines. The acidic residue Glu-395 located at the active site of PPO2 may serve as a general base for deprotonation of monophenolic substrates, which is key to the ortho hydroxylase activity of PO. The structure provides unique insights into the mechanism by which type 3 copper proteins differ in their enzymatic activities, albeit sharing a common active center. A drastic change in electrostatic surface induced on cleavage at Arg-51 allows us to propose a model for localized PPO activation in insects. PMID- 19805073 TI - Identification of functional marker proteins in the mammalian growth cone. AB - Identification of proteins in the mammalian growth cone has the potential to advance our understanding of this critical regulator of neuronal growth and formation of neural circuit; however, to date, only one growth cone marker protein, GAP-43, has been reported. Here, we successfully used a proteomic approach to identify 945 proteins present in developing rat forebrain growth cones, including highly abundant, membrane-associated and actin-associated proteins. Almost 100 of the proteins appear to be highly enriched in the growth cone, as determined by quantitative immunostaining, and for 17 proteins, the results of RNAi suggest a role in axon growth. Most of the proteins we identified have not previously been implicated in axon growth and thus their identification presents a significant step forward, providing marker proteins and candidate neuronal growth-associated proteins. PMID- 19805074 TI - Whole-genome phylogeny of mammals: evolutionary information in genic and nongenic regions. AB - Ten complete mammalian genome sequences were compared by using the "feature frequency profile" (FFP) method of alignment-free comparison. This comparison technique reveals that the whole nongenic portion of mammalian genomes contains evolutionary information that is similar to their genic counterparts--the intron and exon regions. We partitioned the complete genomes of mammals (such as human, chimp, horse, and mouse) into their constituent nongenic, intronic, and exonic components. Phylogenic species trees were constructed for each individual component class of genome sequence data as well as the whole genomes by using standard tree-building algorithms with FFP distances. The phylogenies of the whole genomes and each of the component classes (exonic, intronic, and nongenic regions) have similar topologies, within the optimal feature length range, and all agree well with the evolutionary phylogeny based on a recent large dataset, multispecies, and multigene-based alignment. In the strictest sense, the FFP based trees are genome phylogenies, not species phylogenies. However, the species phylogeny is highly related to the whole-genome phylogeny. Furthermore, our results reveal that the footprints of evolutionary history are spread throughout the entire length of the whole genome of an organism and are not limited to genes, introns, or short, highly conserved, nongenic sequences that can be adversely affected by factors (such as a choice of sequences, homoplasy, and different mutation rates) resulting in inconsistent species phylogenies. PMID- 19805076 TI - Life and death during the Great Depression. AB - Recent events highlight the importance of examining the impact of economic downturns on population health. The Great Depression of the 1930s was the most important economic downturn in the U.S. in the twentieth century. We used historical life expectancy and mortality data to examine associations of economic growth with population health for the period 1920-1940. We conducted descriptive analyses of trends and examined associations between annual changes in health indicators and annual changes in economic activity using correlations and regression models. Population health did not decline and indeed generally improved during the 4 years of the Great Depression, 1930-1933, with mortality decreasing for almost all ages, and life expectancy increasing by several years in males, females, whites, and nonwhites. For most age groups, mortality tended to peak during years of strong economic expansion (such as 1923, 1926, 1929, and 1936-1937). In contrast, the recessions of 1921, 1930-1933, and 1938 coincided with declines in mortality and gains in life expectancy. The only exception was suicide mortality which increased during the Great Depression, but accounted for less than 2% of deaths. Correlation and regression analyses confirmed a significant negative effect of economic expansions on health gains. The evolution of population health during the years 1920-1940 confirms the counterintuitive hypothesis that, as in other historical periods and market economies, population health tends to evolve better during recessions than in expansions. PMID- 19805075 TI - Differing requirements for actin and myosin by plant viruses for sustained intercellular movement. AB - The actin cytoskeleton has been implicated in the intra- and intercellular movement of a growing number of plant and animal viruses. However, the range of viruses influenced by actin for movement and the mechanism of this transport are poorly understood. Here we determine the importance of microfilaments and myosins for the sustained intercellular movement of a group of RNA-based plant viruses. We demonstrate that the intercellular movement of viruses from different genera [tobacco mosaic virus (TMV), potato virus X (PVX), tomato bushy stunt virus (TBSV)], is inhibited by disruption of microfilaments. Surprisingly, turnip vein clearing virus (TVCV), a virus from the same genus as TMV, did not require intact microfilaments for normal spread. To investigate the molecular basis for this difference we compared the subcellular location of GFP fusions to the 126-kDa protein and the homologous 125-kDa protein from TMV and TVCV, respectively. The 126-kDa protein formed numerous large cytoplasmic inclusions associated with microfilaments, whereas the 125-kDa protein formed few small possible inclusions, none associated with microfilaments. The dependence of TMV, PVX, and TBSV on intact microfilaments for intercellular movement led us to investigate the role of myosin motors in this process. Virus-induced gene silencing of the Nicotiana benthamiana myosin XI-2 gene, but not three other myosins, inhibited only TMV movement. These results indicate that RNA viruses have evolved differently in their requirements for microfilaments and the associated myosin motors, in a manner not correlated with predicted phylogeny. PMID- 19805077 TI - Experimental evidence for the conditions necessary to sustain meandering in coarse-bedded rivers. AB - Meandering rivers are common on Earth and other planetary surfaces, yet the conditions necessary to maintain meandering channels are unclear. As a consequence, self-maintaining meandering channels with cutoffs have not been reproduced in the laboratory. Such experimental channels are needed to explore mechanisms controlling migration rate, sinuosity, floodplain formation, and planform morphodynamics and to test theories for wavelength and bend propagation. Here we report an experiment in which meandering with near-constant width was maintained during repeated cutoff and regeneration of meander bends. We found that elevated bank strength (provided by alfalfa sprouts) relative to the cohesionless bed material and the blocking of troughs (chutes) in the lee of point bars via suspended sediment deposition were the necessary ingredients to successful meandering. Varying flood discharge was not necessary. Scaling analysis shows that the experimental meander migration was fast compared to most natural channels. This high migration rate caused nearly all of the bedload sediment to exchange laterally, such that bar growth was primarily dependent on bank sediment supplied from upstream lateral migration. The high migration rate may have contributed to the relatively low sinuosity of 1.19, and this suggests that to obtain much higher sinuosity experiments at this scale may have to be conducted for several years. Although patience is required to evolve them, these experimental channels offer the opportunity to explore several fundamental issues about river morphodynamics. Our results also suggest that sand supply may be an essential control in restoring self-maintaining, actively shifting gravel-bedded meanders. PMID- 19805078 TI - BMP inhibition initiates neural induction via FGF signaling and Zic genes. AB - Neural induction is the process that initiates nervous system development in vertebrates. Two distinct models have been put forward to describe this phenomenon in molecular terms. The default model states that ectoderm cells are fated to become neural in absence of instruction, and do so when bone morphogenetic protein (BMP) signals are abolished. A more recent view implicates a conserved role for FGF signaling that collaborates with BMP inhibition to allow neural fate specification. Using the Xenopus embryo, we obtained evidence that may unite the 2 views. We show that a dominant-negative R-Smad, Smad5-somitabun unlike the other BMP inhibitors used previously-can trigger conversion of Xenopus epidermis into neural tissue in vivo. However, it does so only if FGF activity is uncompromised. We report that this activity may be encoded by FGF4, as its expression is activated upon BMP inhibition, and its knockdown suppresses endogenous, as well as ectopic, neural induction by Smad5-somitabun. Supporting the importance of FGF instructive activity, we report the isolation of 2 immediate early neural targets, zic3 and foxD5a. Conversely, we found that zic1 can be activated by BMP inhibition in the absence of translation. Finally, Zic1 and Zic3 are required together for definitive neural fate acquisition, both in ectopic and endogenous situations. We propose to merge the previous models into a unique one whereby neural induction is controlled by BMP inhibition, which activates directly, and, via FGF instructive activity, early neural regulators such as Zic genes. PMID- 19805079 TI - The human insulin gene is part of a large open chromatin domain specific for human islets. AB - Knowledge of how insulin (INS) gene expression is regulated will lead to better understanding of normal and abnormal pancreatic beta cell function. We have mapped histone modifications over the INS region, coupled with an expression profile, in freshly isolated islets from multiple human donors. Unlike many other human genes, in which active modifications tend to be concentrated within 1 kb around the transcription start site, these marks are distributed over the entire coding region of INS as well. Moreover, a region of approximately 80 kb around the INS gene, which contains the {tyrosine hydroxylase (TH)-(INS)-insulin-like growth factor 2 antisense (IGF2AS)-insulin-like growth factor 2 (IGF2)} gene cluster, unusually is marked by almost uniformly elevated levels of histone acetylation and H3K4 dimethylation, extending both downstream into IGF2 and upstream beyond the TH gene. This is accompanied by islet specific coordinate expression with INS of the neighboring TH and IGF2 genes. The presence of islet specific intergenic transcripts suggests their possible function in the maintenance of this unusual large open chromatin domain. PMID- 19805080 TI - Anoxygenic photosynthesis modulated Proterozoic oxygen and sustained Earth's middle age. AB - Molecular oxygen (O(2)) began to accumulate in the atmosphere and surface ocean ca. 2,400 million years ago (Ma), but the persistent oxygenation of water masses throughout the oceans developed much later, perhaps beginning as recently as 580 550 Ma. For much of the intervening interval, moderately oxic surface waters lay above an oxygen minimum zone (OMZ) that tended toward euxinia (anoxic and sulfidic). Here we illustrate how contributions to primary production by anoxygenic photoautotrophs (including physiologically versatile cyanobacteria) influenced biogeochemical cycling during Earth's middle age, helping to perpetuate our planet's intermediate redox state by tempering O(2) production. Specifically, the ability to generate organic matter (OM) using sulfide as an electron donor enabled a positive biogeochemical feedback that sustained euxinia in the OMZ. On a geologic time scale, pyrite precipitation and burial governed a second feedback that moderated sulfide availability and water column oxygenation. Thus, we argue that the proportional contribution of anoxygenic photosynthesis to overall primary production would have influenced oceanic redox and the Proterozoic O(2) budget. Later Neoproterozoic collapse of widespread euxinia and a concomitant return to ferruginous (anoxic and Fe(2+) rich) subsurface waters set in motion Earth's transition from its prokaryote-dominated middle age, removing a physiological barrier to eukaryotic diversification (sulfide) and establishing, for the first time in Earth's history, complete dominance of oxygenic photosynthesis in the oceans. This paved the way for the further oxygenation of the oceans and atmosphere and, ultimately, the evolution of complex multicellular organisms. PMID- 19805081 TI - Functionally diverse reef-fish communities ameliorate coral disease. AB - Coral reefs, the most diverse of marine ecosystems, currently experience unprecedented levels of degradation. Diseases are now recognized as a major cause of mortality in reef-forming corals and are complicit in phase shifts of reef ecosystems to algal-dominated states worldwide. Even so, factors contributing to disease occurrence, spread, and impact remain poorly understood. Ecosystem resilience has been linked to the conservation of functional diversity, whereas overfishing reduces functional diversity through cascading, top-down effects. Hence, we tested the hypothesis that reefs with trophically diverse reef fish communities have less coral disease than overfished reefs. We surveyed reefs across the central Philippines, including well-managed marine protected areas (MPAs), and found that disease prevalence was significantly negatively correlated with fish taxonomic diversity. Further, MPAs had significantly higher fish diversity and less disease than unprotected areas. We subsequently investigated potential links between coral disease and the trophic components of fish diversity, finding that only the density of coral-feeding chaetodontid butterflyfishes, seldom targeted by fishers, was positively associated with disease prevalence. These previously uncharacterized results are supported by a second large-scale dataset from the Great Barrier Reef. We hypothesize that members of the charismatic reef-fish family Chaetodontidae are major vectors of coral disease by virtue of their trophic specialization on hard corals and their ecological release in overfished areas, particularly outside MPAs. PMID- 19805082 TI - Neural computations underlying action-based decision making in the human brain. AB - Action-based decision making involves choices between different physical actions to obtain rewards. To make such decisions the brain needs to assign a value to each action and then compare them to make a choice. Using fMRI in human subjects, we found evidence for action-value signals in supplementary motor cortex. Separate brain regions, most prominently ventromedial prefrontal cortex, were involved in encoding the expected value of the action that was ultimately taken. These findings differentiate two main forms of value signals in the human brain: those relating to the value of each available action, likely reflecting signals that are a precursor of choice, and those corresponding to the expected value of the action that is subsequently chosen, and therefore reflecting the consequence of the decision process. Furthermore, we also found signals in the dorsomedial frontal cortex that resemble the output of a decision comparator, which implicates this region in the computation of the decision itself. PMID- 19805083 TI - Structures of receptor complexes formed by hemagglutinins from the Asian Influenza pandemic of 1957. AB - The viruses that caused the three influenza pandemics of the twentieth century in 1918, 1957, and 1968 had distinct hemagglutinin receptor binding glycoproteins that had evolved the capacity to recognize human cell receptors. We have determined the structure of the H2 hemagglutinin from the second pandemic, the "Asian Influenza" of 1957. We compare it with the 1918 "Spanish Influenza" hemagglutinin, H1, and the 1968 "Hong Kong Influenza" hemagglutinin, H3, and show that despite its close overall structural similarity to H1, and its more distant relationship to H3, the H2 receptor binding site is closely related to that of H3 hemagglutinin. By analyzing hemagglutinins of potential H2 avian precursors of the pandemic virus, we show that the human receptor can be bound by avian hemagglutinins that lack the human-specific mutations of H2 and H3 pandemic viruses, Gln-226Leu, and Gly-228Ser. We show how Gln-226 in the avian H2 receptor binding site, together with Asn-186, form hydrogen bond networks through bound water molecules to mediate binding to human receptor. We show that the human receptor adopts a very similar conformation in both human and avian hemagglutinin receptor complexes. We also show that Leu-226 in the receptor binding site of human virus hemagglutinins creates a hydrophobic environment near the Sia-1-Gal-2 glycosidic linkage that favors binding of the human receptor and is unfavorable for avian receptor binding. We consider the significance for the development of pandemics, of the existence of avian viruses that can bind to both avian and human receptors. PMID- 19805084 TI - Resolving the distinct stages in erythroid differentiation based on dynamic changes in membrane protein expression during erythropoiesis. AB - Erythropoiesis is the process by which nucleated erythroid progenitors proliferate and differentiate to generate, every second, millions of nonnucleated red cells with their unique discoid shape and membrane material properties. Here we examined the time course of appearance of individual membrane protein components during murine erythropoiesis to throw new light on our understanding of the evolution of the unique features of the red cell membrane. We found that the accumulation of all of the major transmembrane and all skeletal proteins of the mature red blood cell, except actin, accrued progressively during terminal erythroid differentiation. At the same time, and in marked contrast, accumulation of various adhesion molecules decreased. In particular, the adhesion molecule, CD44 exhibited a progressive and dramatic decrease from proerythroblast to reticulocyte; this enabled us to devise a new strategy for distinguishing unambiguously between erythroblasts at successive developmental stages. These findings provide unique insights into the genesis of red cell membrane function during erythroblast differentiation and also offer a means of defining stage specific defects in erythroid maturation in inherited and acquired red cell disorders and in bone marrow failure syndromes. PMID- 19805085 TI - Costly punishment does not always increase cooperation. AB - In a pairwise interaction, an individual who uses costly punishment must pay a cost in order that the opponent incurs a cost. It has been argued that individuals will behave more cooperatively if they know that their opponent has the option of using costly punishment. We examined this hypothesis by conducting two repeated two-player Prisoner's Dilemma experiments, that differed in their payoffs associated to cooperation, with university students from Beijing as participants. In these experiments, the level of cooperation either stayed the same or actually decreased when compared with the control experiments in which costly punishment was not an option. We argue that this result is likely due to differences in cultural attitudes to cooperation and punishment based on similar experiments with university students from Boston that found cooperation did increase with costly punishment. PMID- 19805086 TI - Optical control of zebrafish behavior with halorhodopsin. AB - Expression of halorhodopsin (NpHR), a light-driven microbial chloride pump, enables optical control of membrane potential and reversible silencing of targeted neurons. We generated transgenic zebrafish expressing enhanced NpHR under control of the Gal4/UAS system. Electrophysiological recordings showed that eNpHR stimulation effectively suppressed spiking of single neurons in vivo. Applying light through thin optic fibers positioned above the head of a semi restrained zebrafish larva enabled us to target groups of neurons and to simultaneously test the effect of their silencing on behavior. The photostimulated volume of the zebrafish brain could be marked by subsequent photoconversion of co-expressed Kaede or Dendra. These techniques were used to localize swim command circuitry to a small hindbrain region, just rostral to the commissura infima Halleri. The kinetics of the hindbrain-generated swim command was investigated by combined and separate photo-activation of NpHR and Channelrhodopsin-2 (ChR2), a light-gated cation channel, in the same neurons. Together this "optogenetic toolkit" allows loss-of-function and gain-of-function analyses of neural circuitry at high spatial and temporal resolution in a behaving vertebrate. PMID- 19805087 TI - ADAR1 is required for hematopoietic progenitor cell survival via RNA editing. AB - Adenosine Deaminase Acting on RNA 1 (ADAR1) is an RNA-editing enzyme that converts adenosine to inosine, following RNA transcription. ADAR1's essential role in embryonic development, especially within the hematopoietic lineage, has been demonstrated in knock-out mice. However, a specific role for ADAR1 in adult hematopoietic progenitor cells (HPCs) remains elusive. In this report, we show that ADAR1 is required for survival of differentiating HPCs as opposed to more primitive cells in adult mice by multiple strategies targeting floxed ADAR1 for deletion by Cre recombinase. As a consequence, ADAR1-deficient hematopoietic stem cells (HSCs) were incapable of reconstituting irradiated recipients although being phenotypically present in the recipient bone marrow. While an effect on HSCs cannot be completely ruled out, the preferential effect of ADAR1 absence on HPCs over more primitive hematopoietic cells was consistent with the increased expression of ADAR1 within HPCs, as well as the inability of ADAR1-deficient HPCs to form differentiated colonies and increased apoptotic fraction during ex vivo culture. Moreover, we have obtained direct evidence that ADAR1 functions in HPCs via an RNA-editing dependent mechanism. Therefore, ADAR1 plays an essential role in adult hematopoiesis through its RNA editing activity in HPCs. PMID- 19805088 TI - PNAS takes action regarding breach of NIH embargo policy on a PNAS paper. PMID- 19805089 TI - A wound-induced Wnt expression program controls planarian regeneration polarity. AB - Regeneration requires specification of the identity of new tissues to be made. Whether this process relies only on intrinsic regulative properties of regenerating tissues or whether wound signaling provides input into tissue repatterning is not known. The head-versus-tail regeneration polarity decision in planarians, which requires Wnt signaling, provides a paradigm to study the process of tissue identity specification during regeneration. The Smed-wntP-1 gene is required for regeneration polarity and is expressed at the posterior pole of intact animals. Surprisingly, wntP-1 was expressed at both anterior- and posterior-facing wounds rapidly after wounding. wntP-1 expression was induced by all types of wounds examined, regardless of whether wounding prompted tail regeneration. Regeneration polarity was found to require new expression of wntP 1. Inhibition of the wntP-2 gene enhanced the polarity phenotype due to wntP-1 inhibition, with new expression of wntP-2 in regeneration occurring subsequent to expression of wntP-1 and localized only to posterior-facing wounds. New expression of wntP-2 required wound-induced wntP-1. Finally, wntP-1 and wntP-2 expression changes occurred even in the absence of neoblast stem cells, which are required for regeneration, suggesting that the role of these genes in polarity is independent of and instructive for tail formation. These data indicate that wound induced input is involved in resetting the normal polarized features of the body axis during regeneration. PMID- 19805090 TI - Membrane domains and flagellar pocket boundaries are influenced by the cytoskeleton in African trypanosomes. AB - A key feature of immune evasion for African trypanosomes is the functional specialization of their surface membrane in an invagination known as the flagellar pocket (FP), the cell's sole site of endocytosis and exocytosis. The FP membrane is biochemically distinct yet continuous with those of the cell body and the flagellum. The structural features maintaining this individuality are not known, and we lack a clear understanding of how extracellular components gain access to the FP. Here, we have defined domains and boundaries on these surface membranes and identified their association with internal cytoskeletal features. The FP membrane appears largely homogeneous and uniformly involved in endocytosis. However, when endocytosis is blocked, receptor-mediated and fluid phase endocytic markers accumulate specifically on membrane associated with four specialized microtubules in the FP region. These microtubules traverse a distinct boundary and associate with a channel that connects the FP lumen to the extracellular space, suggesting that the channel is the major transport route into the FP. PMID- 19805091 TI - Kinesin's step dissected with single-motor FRET. AB - The motor protein Kinesin-1 drives intracellular transport along microtubules, with each of its two motor domains taking 16-nm steps in a hand-over-hand fashion. The way in which a single-motor domain moves during a step is unknown. Here, we use Forster resonance energy transfer (FRET) between fluorescent labels on both motor domains of a single kinesin. This approach allows us to resolve the relative distance between the motor domains and their relative orientation, on the submillisecond timescale, during processive stepping. We observe transitions between high and low FRET values for certain kinesin constructs, depending on the location of the labels. These results reveal that, during a step, a kinesin motor domain dwells in a well-defined intermediate position for approximately 3 ms. PMID- 19805092 TI - A selective contribution of the RIG-I-like receptor pathway to type I interferon responses activated by cytosolic DNA. AB - The activation of the innate immune responses by DNA exposed within the cytosol has gained much attention and, in this context, several cytosolic DNA sensors have been identified. However, previous studies revealed the operation of redundant and complex mechanisms and it still remains to be clarified how the DNA mediated evocation of diverse innate immune responses can be achieved. Here we show that two RIG-I-like receptors (RLRs), RIG-I and MDA5, known as cytosolic RNA receptors, nonredundantly function as cytosolic DNA receptors that lead to the selective activation of type I IFN genes. We demonstrate that overexpression of otherwise IFN-inducible RIG-I or MDA5 in IFN signal-deficient cells results in a marked enhancement of type I IFN gene induction upon cytosolic DNA stimulation, while in their absence the induction is impaired. Interestingly, the DNA-mediated induction of other cytokine genes was barely affected by the absence of RLRs. Indeed, unlike the RNA-RLR pathway that activates the transcription factors IRF3 and NF-kappaB, the DNA-RLR pathway is primarily responsible for the IRF3 activation critical for type I IFN gene transcription, illustrating a deliberate divergence of the DNA signaling pathways. Expectedly, the RLR pathway also contributes to intricate innate immune responses against infection by a DNA virus. Our study may provide insights into the complexity of host defense mechanisms that thwart immune evasion by DNA-containing pathogens. PMID- 19805093 TI - Appraising the apoptotic mimicry model and the role of phospholipids for poxvirus entry. AB - Entry of vaccinia virus (VACV) into cells occurs by fusion with the plasma membrane and via a low pH-dependent endosomal pathway, presumably involving unidentified cellular receptors. In addition to approximately 25 viral proteins, the membrane of VACV mature virions contains several phospholipids including phosphatidylserine (PS). A recent model posits that PS flags virions as apoptotic debris to activate a common cellular uptake pathway to gain cell entry, perhaps through an interaction with a PS-specific cell surface receptor. To evaluate the apoptotic mimicry model, we reconstituted the membrane of detergent-extracted virions with several different phospholipids. Although the ability of the L stereoisomer of PS to reconstitute infectivity was confirmed, the nonbiologically relevant D-stereoisomer of PS, and phosphatidylglycerol, which are not normally present in the virion membrane, functioned as well. Regardless of which phospholipid reconstituted infectivity, virus entry was inhibited by a neutralizing monoclonal antibody to a virion surface protein and by the drugs blebbistatin and bafilomycin A1, suggesting that in each case virus uptake was specific and occurred by a similar mechanism involving macropinocytosis and a low pH endocytic pathway. Lipid-reconstituted and nonreconstituted, membrane extracted virions were equally capable of binding to cells. However, the physical association of phospholipids with virus particles during membrane reconstitution correlated directly with rescue of particle infectivity and cell entry capability. Our results support a role for PS in poxvirus entry, but demonstrate that other phospholipids, not known to signal uptake of apoptotic debris, can function similarly. PMID- 19805094 TI - Targeting lymphotoxin-mediated negative selection to prevent prostate cancer in mice with genetic predisposition. AB - The identification of individuals genetically susceptible to cancer calls for preventive measures to minimize the cancer risk in these high-risk populations. Immune prevention is made necessary by the anticipated health threat, but lack of enough high-affinity T cells against tumor-associated antigens and the unpredictability of tumor antigens make antigen-based immune prevention untenable for cancer. To address this issue, we explored a non-antigen-based cancer immune prevention strategy using the transgenic adenocarcinoma of mouse prostate model that spontaneously develops prostate cancer with 100% penetrance. We show that targeted mutation of the lymphotoxin alpha (LTalpha) gene efficiently rescued tumor-reactive T cells, drastically reduced cancer incidence, and almost completely ablated metastasis. Remarkably, short-term treatments with the fusion protein consisting of constant region of IgG and extracellular domain of lymphotoxin beta receptor (LTbetaRIg) interrupted clonal deletion, reduced the size of the primary cancer, and completely prevented metastasis later in life. Our data demonstrated the value of non-antigen-based immune prevention for those with a genetic predisposition to cancer. PMID- 19805095 TI - A human IL10 BAC transgene reveals tissue-specific control of IL-10 expression and alters disease outcome. AB - Interleukin (IL)-10 is an immunoregulatory cytokine that is produced by diverse cell populations. Studies in mice suggest that the cellular source of IL-10 is a key determinant in various disease pathologies, yet little is known regarding the control of tissue-specific human IL-10 expression. To assess cell type-specific human IL-10 regulation, we created a human IL-10 transgenic mouse with a bacterial artificial chromosome (hIL10BAC) in which the IL10 gene is positioned centrally. Since human IL-10 is biologically active in the mouse, we could examine the in vivo capacity of tissue-specific human IL-10 expression to recapitulate IL-10-dependent phenotypes by reconstituting Il10(-/-) mice (Il10(-/ )/hIL10BAC). In response to LPS, Il10(-/-)/hIL10BAC mice proficiently regulate IL 10-target genes and normalize sensitivity to LPS toxicity via faithful human IL 10 expression from macrophages and dendritic cells. However, in the Leishmania donovani model of pathogen persistence, Il10(-/-)/hIL10BAC mice did not develop the characteristic IL-10(+)IFN-gamma(+)CD4 T cell subset thought to mediate persistence and, like Il10(-/-) mice, cleared the parasites. Furthermore, the IL 10-promoting cytokine IL-27 failed to regulate transgenic human IL-10 production in CD4(+) T cells in vitro which together suggests that the hIL10BAC encodes for weak T cell-specific IL-10 expression. Thus, the hIL10BAC mouse is a model of human gene structure and function revealing tissue-specific regulatory requirements for IL-10 expression which impacts disease outcomes. PMID- 19805096 TI - Discovery and validation of colonic tumor-associated proteins via metabolic labeling and stable isotopic dilution. AB - The unique biology of a neoplasm is reflected by its distinct molecular profile compared with normal tissue. To understand tumor development better, we have undertaken a quantitative proteomic search for abnormally expressed proteins in colonic tumors from Apc(Min/+) (Min) mice. By raising pairs of Min and wild-type mice on diets derived from natural-abundance or (15)N-labeled algae, we used metabolic labeling to compare protein levels in colonic tumor versus normal tissue. Because metabolic labeling allows internal control throughout sample preparation and analysis, technical error is minimized as compared with in vitro labeling. Several proteins displayed altered expression, and a subset was validated via stable isotopic dilution using synthetic peptide standards. We also compared gene and protein expression among tumor and nontumor tissue, revealing limited correlation. This divergence was especially pronounced for species showing biological change, highlighting the complementary perspectives provided by transcriptomics and proteomics. Our work demonstrates the power of metabolic labeling combined with stable isotopic dilution as an integrated strategy for the identification and validation of differentially expressed proteins using rodent models of human disease. PMID- 19805097 TI - A coiled-coil motif that sequesters ions to the hydrophobic core. AB - Most core residues of coiled coils are hydrophobic. Occasional polar residues are thought to lower stability, but impart structural specificity. The coiled coils of trimeric autotransporter adhesins (TAAs) are conspicuous for their large number of polar residues in position d of the core, which often leads to their prediction as natively unstructured regions. The most frequent residue, asparagine (N@d), can occur in runs of up to 19 consecutive heptads, frequently in the motif [I/V]xxNTxx. In the Salmonella TAA, SadA, the core asparagines form rings of interacting residues with the following threonines, grouped around a central anion. This conformation is observed generally in N@d layers from trimeric coiled coils of known structure. Attempts to impose a different register on the motif show that the asparagines orient themselves specifically into the core, even against conflicting information from flanking domains. When engineered into the GCN4 leucine zipper, N@d layers progressively destabilized the structure, but zippers with 3 N@d layers still folded at high concentration. We propose that N@d layers maintain the coiled coils of TAAs in a soluble, export competent state during autotransport through the outer membrane. More generally, we think that polar motifs that are both periodic and conserved may often reflect special folding requirements, rather than an unstructured state of the mature proteins. PMID- 19805098 TI - Molecules, morphology, and ecology indicate a recent, amphibious ancestry for echidnas. AB - The semiaquatic platypus and terrestrial echidnas (spiny anteaters) are the only living egg-laying mammals (monotremes). The fossil record has provided few clues as to their origins and the evolution of their ecological specializations; however, recent reassignment of the Early Cretaceous Teinolophos and Steropodon to the platypus lineage implies that platypuses and echidnas diverged >112.5 million years ago, reinforcing the notion of monotremes as living fossils. This placement is based primarily on characters related to a single feature, the enlarged mandibular canal, which supplies blood vessels and dense electrosensory receptors to the platypus bill. Our reevaluation of the morphological data instead groups platypus and echidnas to the exclusion of Teinolophos and Steropodon and suggests that an enlarged mandibular canal is ancestral for monotremes (partly reversed in echidnas, in association with general mandibular reduction). A multigene evaluation of the echidna-platypus divergence using both a relaxed molecular clock and direct fossil calibrations reveals a recent split of 19-48 million years ago. Platypus-like monotremes (Monotrematum) predate this divergence, indicating that echidnas had aquatically foraging ancestors that reinvaded terrestrial ecosystems. This ecological shift and the associated radiation of echidnas represent a recent expansion of niche space despite potential competition from marsupials. Monotremes might have survived the invasion of marsupials into Australasia by exploiting ecological niches in which marsupials are restricted by their reproductive mode. Morphology, ecology, and molecular biology together indicate that Teinolophos and Steropodon are basal monotremes rather than platypus relatives, and that living monotremes are a relatively recent radiation. PMID- 19805099 TI - Structural basis for Ca2+-independence and activation by homodimerization of tomato subtilase 3. AB - Subtilases are serine proteases found in Archae, Bacteria, yeasts, and higher eukaryotes. Plants possess many more of these subtilisin-like endopeptidases than animals, e.g., 56 identified genes in Arabidopsis compared with only 9 in humans, indicating important roles for subtilases in plant biology. We report the first structure of a plant subtilase, SBT3 from tomato, in the active apo form and complexed with a chloromethylketone (cmk) inhibitor. The domain architecture comprises an N-terminal protease domain displaying a 132 aa protease-associated (PA) domain insertion and a C-terminal seven-stranded jelly-roll fibronectin (Fn) III-like domain. We present the first structural evidence for an explicit function of PA domains in proteases revealing a vital role in the homo dimerization of SBT3 and in enzyme activation. Although Ca(2+)-binding sites are conserved and critical for stability in other subtilases, SBT3 was found to be Ca(2+)-free and its thermo stability is Ca(2+)-independent. PMID- 19805100 TI - Optimizing rotary processes in synthetic molecular motors. AB - We deal with the issue of quantifying and optimizing the rotation dynamics of synthetic molecular motors. For this purpose, the continuous four-stage rotation behavior of a typical light-activated molecular motor was measured in detail. All reaction constants were determined empirically. Next, we developed a Markov model that describes the full motor dynamics mathematically. We derived expressions for a set of characteristic quantities, i.e., the average rate of quarter rotations or "velocity," V, the spread in the average number of quarter rotations, D, and the dimensionless Peclet number, Pe = V/D. Furthermore, we determined the rate of full, four-step rotations (Omega(eff)), from which we derived another dimensionless quantity, the "rotational excess," r.e. This quantity, defined as the relative difference between total forward (Omega(+)) and backward (Omega(-)) full rotations, is a good measure of the unidirectionality of the rotation process. Our model provides a pragmatic tool to optimize motor performance. We demonstrate this by calculating V, D, Pe, Omega(eff), and r.e. for different rates of thermal versus photochemical energy input. We find that for a given light intensity, an optimal temperature range exists in which the motor exhibits excellent efficiency and unidirectional behavior, above or below which motor performance decreases. PMID- 19805101 TI - Polymer beacons for luminescence and magnetic resonance imaging of DNA delivery. AB - The delivery of nucleic acids with polycations offers tremendous potential for developing highly specific treatments for various therapeutic targets. Although materials have been developed and studied for polynucleotide transfer, the biological mechanisms and fate of the synthetic vehicle has remained elusive due to the limitations with current labeling technologies. Here, we have developed polymer beacons that allow the delivery of nucleic acids to be visualized at different biological scales. The polycations have been designed to contain repeated oligoethyleneamines, for binding and compacting nucleic acids into nanoparticles, and lanthanide (Ln) chelates [either luminescent europium (Eu(3+)) or paramagnetic gadolinium (Gd(3+))]. The chelated Lns allow the visualization of the delivery vehicle both on the nm/microm scale via microscopy and on the sub-mm scale via MRI. We demonstrate that these delivery beacons effectively bind and compact plasmid (p)DNA into nanoparticles and protect nucleic acids from nuclease damage. These delivery beacons efficiently deliver pDNA into cultured cells and do not exhibit toxicity. Micrographs of cultured cells exposed to the nanoparticle complexes formed with fluorescein-labeled pDNA and the europium chelated polymers reveal effective intracellular imaging of the delivery process. MRI of bulk cells exposed to the complexes formulated with pDNA and the gadolinium-chelated structures show bright image contrast, allowing visualization of effective intracellular delivery on the tissue-scale. Because of their versatility, these delivery beacons posses remarkable potential for tracking and understanding nucleic acid transfer in vitro, and have promise as in vivo theranostic agents. PMID- 19805102 TI - Universal architecture of bacterial chemoreceptor arrays. AB - Chemoreceptors are key components of the high-performance signal transduction system that controls bacterial chemotaxis. Chemoreceptors are typically localized in a cluster at the cell pole, where interactions among the receptors in the cluster are thought to contribute to the high sensitivity, wide dynamic range, and precise adaptation of the signaling system. Previous structural and genomic studies have produced conflicting models, however, for the arrangement of the chemoreceptors in the clusters. Using whole-cell electron cryo-tomography, here we show that chemoreceptors of different classes and in many different species representing several major bacterial phyla are all arranged into a highly conserved, 12-nm hexagonal array consistent with the proposed "trimer of dimers" organization. The various observed lengths of the receptors confirm current models for the methylation, flexible bundle, signaling, and linker sub-domains in vivo. Our results suggest that the basic mechanism and function of receptor clustering is universal among bacterial species and was thus conserved during evolution. PMID- 19805103 TI - Microwell-mediated control of embryoid body size regulates embryonic stem cell fate via differential expression of WNT5a and WNT11. AB - Recently, various approaches for controlling the embryonic stem (ES) cell microenvironment have been developed for regulating cellular fate decisions. It has been reported that the lineage specific differentiation could be affected by the size of ES cell colonies and embryoid bodies (EBs). However, much of the underlying biology has not been well elucidated. In this study, we used microengineered hydrogel microwells to direct ES cell differentiation and determined the role of WNT signaling pathway in directing the differentiation. This was accomplished by forming ES cell aggregates within microwells to form different size EBs. We determined that cardiogenesis was enhanced in larger EBs (450 microm in diameter), and in contrast, endothelial cell differentiation was increased in smaller EBs (150 microm in diameter). Furthermore, we demonstrated that the EB-size mediated differentiation was driven by differential expression of WNTs, particularly noncanonical WNT pathway, according to EB size. The higher expression of WNT5a in smaller EBs enhanced endothelial cell differentiation. In contrast, the increased expression of WNT11 enhanced cardiogenesis. This was further validated by WNT5a-siRNA transfection assay and the addition of recombinant WNT5a. Our data suggest that EB size could be an important parameter in ES cell fate specification via differential gene expression of members of the noncanonical WNT pathway. Given the size-dependent response of EBs to differentiate to endothelial and cardiac lineages, hydrogel microwell arrays could be useful for directing stem cell fates and studying ES cell differentiation in a controlled manner. PMID- 19805104 TI - Ecology and the ratchet of events: climate variability, niche dimensions, and species distributions. AB - Climate change in the coming centuries will be characterized by interannual, decadal, and multidecadal fluctuations superimposed on anthropogenic trends. Predicting ecological and biogeographic responses to these changes constitutes an immense challenge for ecologists. Perspectives from climatic and ecological history indicate that responses will be laden with contingencies, resulting from episodic climatic events interacting with demographic and colonization events. This effect is compounded by the dependency of environmental sensitivity upon life-stage for many species. Climate variables often used in empirical niche models may become decoupled from the proximal variables that directly influence individuals and populations. Greater predictive capacity, and more-fundamental ecological and biogeographic understanding, will come from integration of correlational niche modeling with mechanistic niche modeling, dynamic ecological modeling, targeted experiments, and systematic observations of past and present patterns and dynamics. PMID- 19805105 TI - A frequent kinase domain mutation that changes the interaction between PI3Kalpha and the membrane. AB - Mutations in oncogenes often promote tumorigenesis by changing the conformation of the encoded proteins, thereby altering enzymatic activity. The PIK3CA oncogene, which encodes p110alpha, the catalytic subunit of phosphatidylinositol 3-kinase alpha (PI3Kalpha), is one of the two most frequently mutated oncogenes in human cancers. We report the structure of the most common mutant of p110alpha in complex with two interacting domains of its regulatory partner (p85alpha), both free and bound to an inhibitor (wortmannin). The N-terminal SH2 (nSH2) domain of p85alpha is shown to form a scaffold for the entire enzyme complex, strategically positioned to communicate extrinsic signals from phosphopeptides to three distinct regions of p110alpha. Moreover, we found that Arg-1047 points toward the cell membrane, perpendicular to the orientation of His-1047 in the WT enzyme. Surprisingly, two loops of the kinase domain that contact the cell membrane shift conformation in the oncogenic mutant. Biochemical assays revealed that the enzymatic activity of the p110alpha His1047Arg mutant is differentially regulated by lipid membrane composition. These structural and biochemical data suggest a previously undescribed mechanism for mutational activation of a kinase that involves perturbation of its interaction with the cellular membrane. PMID- 19805106 TI - Photolysis of iron-siderophore chelates promotes bacterial-algal mutualism. AB - Marine microalgae support world fisheries production and influence climate through various mechanisms. They are also responsible for harmful blooms that adversely impact coastal ecosystems and economies. Optimal growth and survival of many bloom-forming microalgae, including climatically important dinoflagellates and coccolithophores, requires the close association of specific bacterial species, but the reasons for these associations are unknown. Here, we report that several clades of Marinobacter ubiquitously found in close association with dinoflagellates and coccolithophores produce an unusual lower-affinity dicitrate siderophore, vibrioferrin (VF). Fe-VF chelates undergo photolysis at rates that are 10-20 times higher than siderophores produced by free-living marine bacteria, and unlike the latter, the VF photoproduct has no measurable affinity for iron. While both an algal-associated bacterium and a representative dinoflagellate partner, Scrippsiella trochoidea, used iron from Fe-VF chelates in the dark, in situ photolysis of the chelates in the presence of attenuated sunlight increased bacterial iron uptake by 70% and algal uptake by >20-fold. These results suggest that the bacteria promote algal assimilation of iron by facilitating photochemical redox cycling of this critical nutrient. Also, binary culture experiments and genomic evidence suggest that the algal cells release organic molecules that are used by the bacteria for growth. Such mutualistic sharing of iron and fixed carbon has important implications toward our understanding of the close beneficial interactions between marine bacteria and phytoplankton, and the effect of these interactions on algal blooms and climate. PMID- 19805107 TI - Inhibition of Hsp90 via 17-DMAG induces apoptosis in a p53-dependent manner to prevent medulloblastoma. AB - Elevated expression of HSP90 is observed in many tumor types and is associated with a limited clinical response. Targeting HSP90 using inhibitors such as 17 DMAG (17-desmethoxy-17-N,N-dimethylaminoethylaminogeldanamycin) has shown limited therapeutic success. HSP90 regulates the function of several proteins implicated in tumorigenesis although the precise mechanism through which 17-DMAG regulates tumor cell survival remains unclear. We observed a requirement for p53 in mediating 17-DMAG-induced cell death. The sensitivity of primary mouse embryonic fibroblasts and tumor cells to 17-DMAG-induced apoptosis depended on the p53 status. Wild-type MEFs underwent 17-DMAG-induced caspase-dependent cell death, whilst those lacking p53 failed to do so. Interestingly p53-dependent cell death occurred independently of Atm or Arf. Primary tumor cells derived from two models of murine medulloblastoma (Ptch1(+/-);Ink4c(-/-) and p53(FL/FL);Nestin-Cre(+); Ink4c(-/-)) that retain and lack p53 function, respectively, displayed a dependence on functional p53 to engage 17-DMAG-induced apoptosis. Strikingly, 17 DMAG treatment in an allograft model of Ptch1(+/-);Ink4c(-/-) but not p53(FL/FL);Nestin-Cre(+); Ink4c(-/-) tumor cells prevented tumor growth in vivo. Our data suggest that p53 status is a likely predictor of the sensitivity of tumors to 17-DMAG. PMID- 19805108 TI - Orthogonal analytical approaches to detect potential contaminants in heparin. AB - Heparin is a widely used anticoagulant and antithrombotic agent. Recently, a contaminant, oversulfated chondroitin sulfate (OSCS), was discovered within heparin preparations. The presence of OSCS within heparin likely led to clinical manifestations, most prevalently, hypotension and abdominal pain leading to the deaths of several dozens of patients. Given the biological effects of OSCS, one continuing item of concern is the ability for existing methods to identify other persulfonated polysaccharide compounds that would also have anticoagulant activity and would likely elicit a similar activation of the contact system. To complete a more extensive analysis of the ability for NMR and capillary electrophoresis (CE) to capture a broader array of potential contaminants within heparin, we completed a systematic study of NMR, both mono- and bidimensional, and CE to detect both various components of sidestream heparin and their persulfonated derivatives. We show that given the complexity of heparin samples, and the requirement to ensure their purity and safety, use of orthogonal analytical techniques is effective at detecting an array of potential contaminants that could be present. PMID- 19805109 TI - A strategy for blood biomarker amplification and localization using ultrasound. AB - Blood biomarkers have significant potential applications in early detection and management of various diseases, including cancer. Most biomarkers are present in low concentrations in blood and are difficult to discriminate from noise. Furthermore, blood measurements of a biomarker do not provide information about the location(s) where it is produced. We hypothesize a previously undescribed strategy to increase the concentration of biomarkers in blood as well as localize the source of biomarker signal using ultrasound energy directly applied to tumor cells. We test and validate our hypothesis in cell culture experiments and mouse tumor xenograft models using the human colon cancer cell line LS174T, while measuring the biomarker carcinoembryonic antigen (CEA) before and after the use of ultrasound to liberate the biomarker from the tumor cells. The results demonstrate that the application of low-frequency ultrasound to tumor cells causes a significant release of tumor biomarker, which can be measured in the blood. Furthermore, we establish that this release is specific to the direct application of the ultrasound to the tumor, enabling a method for localization of biomarker production. This work shows that it is possible to use ultrasound to amplify and localize the source of CEA levels in blood of tumor-bearing mice and will allow for a previously undescribed way to determine the presence and localization of disease more accurately using a relatively simple and noninvasive strategy. PMID- 19805110 TI - Catalytic activities of NifEN: implications for nitrogenase evolution and mechanism. AB - NifEN is a key player in the biosynthesis of nitrogenase MoFe protein. It not only shares a considerable degree of sequence homology with the MoFe protein, but also contains clusters that are homologous to those found in the MoFe protein. Here we present an investigation of the catalytic activities of NifEN. Our data show that NifEN is catalytically competent in acetylene (C(2)H(2)) and azide (N(3)(-)) reduction, yet unable to reduce dinitrogen (N(2)) or evolve hydrogen (H(2)). Upon turnover, C(2)H(2) gives rise to an additional S = 1/2 signal, whereas N(3)(-) perturbs the signal originating from the NifEN-associated FeMoco homolog. Combined biochemical and spectroscopic studies reveal that N(3)(-) can act as either an inhibitor or an activator for the binding and/or reduction of C(2)H(2), while carbon monoxide (CO) is a potent inhibitor for the binding and/or reduction of both N(3)(-) and C(2)H(2). Taken together, our results suggest that NifEN is a catalytic homolog of MoFe protein; however, it is only a "skeleton" version of the MoFe protein, as its associated clusters are simpler in structure and less versatile in function, which, in turn, may account for its narrower range of substrates and lower activities of substrate reduction. The resemblance of NifEN to MoFe protein in catalysis points to a plausible, sequential appearance of the two proteins in nitrogenase evolution. More importantly, the discrepancy between the two systems may provide useful insights into nitrogenase mechanism and allow reconstruction of a fully functional nitrogenase from the "skeleton" enzyme, NifEN. PMID- 19805111 TI - Direct measurements of kinesin torsional properties reveal flexible domains and occasional stalk reversals during stepping. AB - Kinesin is a homodimeric motor with two catalytic heads joined to a stalk via short neck linkers (NLs). We measured the torsional properties of single recombinant molecules by tracking the thermal angular motions of fluorescently labeled beads bound to the C terminus of the stalk. When kinesin heads were immobilized on microtubules (MTs) under varied nucleotide conditions, we observed bounded or unbounded angular diffusion, depending on whether one or both heads were attached to the MT. Free rotation implies that NLs act as swivels. From data on constrained diffusion, we conclude that the coiled-coil stalk domains are approximately 30-fold stiffer than its flexible "hinge" regions. Surprisingly, while tracking processive kinesin motion at low ATP concentrations, we observed occasional abrupt reversals in the directional orientations of the stalk. Our results impose constraints on kinesin walking models and suggest a role for rotational freedom in cargo transport. PMID- 19805113 TI - Sufficient symmetry conditions for Topological Quantum Order. AB - We prove sufficient conditions for Topological Quantum Order at zero and finite temperatures. The crux of the proof hinges on the existence of low-dimensional Gauge-Like Symmetries, thus providing a unifying framework based on a symmetry principle. These symmetries may be actual invariances of the system, or may emerge in the low-energy sector. Prominent examples of Topological Quantum Order display Gauge-Like Symmetries. New systems exhibiting such symmetries include Hamiltonians depicting orbital-dependent spin exchange and Jahn-Teller effects in transition metal orbital compounds, short-range frustrated Klein spin models, and p+ip superconducting arrays. We analyze the physical consequences of Gauge-Like Symmetries (including topological terms and charges) and show the insufficiency of the energy spectrum, topological entanglement entropy, maximal string correlators, and fractionalization in establishing Topological Quantum Order. General symmetry considerations illustrate that not withstanding spectral gaps, thermal fluctuations may impose restrictions on suggested quantum computing schemes. Our results allow us to go beyond standard topological field theories and engineer systems with Topological Quantum Order. PMID- 19805112 TI - CCR7-mediated migration of developing thymocytes to the medulla is essential for negative selection to tissue-restricted antigens. AB - Immature double-positive thymocytes are generated in the thymic cortex, and on positive selection, are induced to differentiate into mature single-positive thymocytes and relocate to the medulla. CCR7 is pivotal for cortex-to-medulla migration of positively selected thymocytes, and CCR7-mediated migration to the medulla is essential for establishing central tolerance, thereby, preventing tissue-specific autoimmunity. However, it was unclear how CCR7-mediated migration to the medulla affects the establishment of self-tolerance. Here, we show that the deletion of thymocytes specific for insulin-promoter-driven tissue-restricted antigens (TRAs) is significantly impaired in CCR7- or CCR7-ligand-deficient mice. These results indicate that CCR7-mediated migration to the medulla contributes to the negative selection of TRA-reactive thymocytes. PMID- 19805114 TI - Suppression of retinal degeneration in Drosophila by stimulation of ER-associated degradation. AB - Mutations in the rhodopsin gene that disrupt the encoded protein's folding properties are a major cause of autosomal dominant retinitis pigmentosa (ADRP). This disease is faithfully modeled in Drosophila where similar mutations in the ninaE gene, encoding rhodopsin-1 (Rh-1), cause ER stress and dominantly trigger age-related retinal degeneration. In addition, mutant flies bearing certain ninaE alleles have dramatically reduced Rh-1 protein levels, but the underlying mechanism for this reduction and significance of its contribution to the ADRP phenotype remains unclear. To address this question, we specifically analyzed the role of Drosophila genes homologous to the known yeast and animal regulators of the ER-associated degradation (ERAD) pathway, a process that reduces levels of misfolded proteins in the ER through proteasomal degradation. We found that loss of-function of these putative ERAD factors resulted in increased levels of Rh-1 in ninaE mutant flies. Conversely, in an ER stress assay where mutant or wild type Rh-1 were overexpressed in developing imaginal discs beyond the ER protein folding capacity of those cells, co-expression of certain ERAD factors was sufficient to reduce Rh-1 protein levels and to completely suppress ER stress reporter activation. Significantly, those ERAD factors that specifically reduced misfolded Rh-1 in the imaginal disc assay also delayed age-related retinal degeneration caused by an endogenous ninaE allele, indicating that ERAD acts as a protective mechanism against retinal degeneration in the Drosophila model for ADRP. These results suggest that manipulation of ERAD may serve as a powerful therapeutic strategy against a number of diseases associated with ER stress. PMID- 19805115 TI - Anomalous properties of the acoustic excitations in glasses on the mesoscopic length scale. AB - The low-temperature thermal properties of dielectric crystals are governed by acoustic excitations with large wavelengths that are well described by plane waves. This is the Debye model, which rests on the assumption that the medium is an elastic continuum, holds true for acoustic wavelengths large on the microscopic scale fixed by the interatomic spacing, and gradually breaks down on approaching it. Glasses are characterized as well by universal low-temperature thermal properties that are, however, anomalous with respect to those of the corresponding crystalline phases. Related universal anomalies also appear in the low-frequency vibrational density of states and, despite a longstanding debate, remain poorly understood. By using molecular dynamics simulations of a model monatomic glass of extremely large size, we show that in glasses the structural disorder undermines the Debye model in a subtle way: The elastic continuum approximation for the acoustic excitations breaks down abruptly on the mesoscopic, medium-range-order length scale of approximately 10 interatomic spacings, where it still works well for the corresponding crystalline systems. On this scale, the sound velocity shows a marked reduction with respect to the macroscopic value. This reduction turns out to be closely related to the universal excess over the Debye model prediction found in glasses at frequencies of approximately 1 THz in the vibrational density of states or at temperatures of approximately 10 K in the specific heat. PMID- 19805116 TI - Structural basis of activation-dependent binding of ligand-mimetic antibody AL-57 to integrin LFA-1. AB - The activity of integrin LFA-1 (alpha(L)beta(2)) to its ligand ICAM-1 is regulated through the conformational changes of its ligand-binding domain, the I domain of alpha(L) chain, from an inactive, low-affinity closed form (LA), to an intermediate-affinity form (IA), and then finally, to a high-affinity open form (HA). A ligand-mimetic human monoclonal antibody AL-57 (activated LFA-1 clone 57) was identified by phage display to specifically recognize the affinity upregulated I domain. Here, we describe the crystal structures of the Fab fragment of AL-57 in complex with IA, as well as in its unligated form. We discuss the structural features conferring AL-57's strong selectivity for the high affinity, open conformation of the I domain. The AL-57-binding site overlaps the ICAM-1 binding site on the I domain. Furthermore, an antibody Asp mimics an ICAM Glu by forming a coordination to the metal-ion dependent adhesion site (MIDAS). The structure also reveals better shape complementarity and a more hydrophobic interacting interface in AL-57 binding than in ICAM-1 binding. The results explain AL-57's antagonistic mimicry of LFA-1's natural ligands, the ICAM molecules. PMID- 19805117 TI - A genomic screen identifies TYRO3 as a MITF regulator in melanoma. AB - Malignant melanoma is the most aggressive form of cutaneous carcinoma, accounting for 75% of all deaths caused by skin cancers. Microphthalmia-associated transcription factor (MITF) is a master gene regulating melanocyte development and functions as a "lineage addiction" oncogene in malignant melanoma. We have identified the receptor protein tyrosine kinase TYRO3 as an upstream regulator of MITF expression by a genome-wide gain-of-function cDNA screen and show that TYRO3 induces MITF-M expression in a SOX10-dependent manner in melanoma cells. Expression of TYRO3 is significantly elevated in human primary melanoma tissue samples and melanoma cell lines and correlates with MITF-M mRNA levels. TYRO3 overexpression bypasses BRAF(V600E)-induced senescence in primary melanocytes, inducing transformation of non-tumorigenic cell lines. Furthermore, TYRO3 knockdown represses cellular proliferation and colony formation in melanoma cells, and sensitizes them to chemotherapeutic agent-induced apoptosis; TYRO3 knockdown in melanoma cells also inhibits tumorigenesis in vivo. Taken together, these data indicate that TYRO3 may serve as a target for the development of therapeutic agents for melanoma. PMID- 19805118 TI - Image-based multiscale modeling predicts tissue-level and network-level fiber reorganization in stretched cell-compacted collagen gels. AB - The mechanical environment plays an important role in cell signaling and tissue homeostasis. Unraveling connections between externally applied loads and the cellular response is often confounded by extracellular matrix (ECM) heterogeneity. Image-based multiscale models provide a foundation for examining the fine details of tissue behavior, but they require validation at multiple scales. In this study, we developed a multiscale model that captured the anisotropy and heterogeneity of a cell-compacted collagen gel subjected to an off axis hold mechanical test and subsequently to biaxial extension. In both the model and experiments, the ECM reorganized in a nonaffine and heterogeneous manner that depended on multiscale interactions between the fiber networks. Simulations predicted that tensile and compressive fiber forces were produced to accommodate macroscopic displacements. Fiber forces in the simulation ranged from -11.3 to 437.7 nN, with a significant fraction of fibers under compression (12.1% during off-axis stretch). The heterogeneous network restructuring predicted by the model serves as an example of how multiscale modeling techniques provide a theoretical framework for understanding relationships between ECM structure and tissue-level mechanical properties and how microscopic fiber rearrangements could lead to mechanotransductive cell signaling. PMID- 19805119 TI - Impaired tumor growth, metastasis, angiogenesis and wound healing in annexin A1 null mice. AB - Despite 2 decades of research, no clear function for annexin A1 (AnxA1) has been established. Using AnxA1-KO mice, we show that tumor growth and metastasis are significantly decreased, whereas rodent survival and tumor necrosis are greatly increased when tumors grow in AnxA1-KO mice. Systems analysis of gene expression in these tumors specifically implicates 2 related vascular functions, angiogenesis and wound healing, in this impairment. Both tumor vascular development and wound healing are greatly retarded in KO tissues. Aortic ring assays reveal induced AnxA1 expression on sprouting endothelial cells of normal mice whereas KO aortas exhibit impaired endothelial cell sprouting that is rescued by adenoviral expression of AnxA1. Key differences in specific gene regulation may define new molecular pathways mediating angiogenesis, including a reset profile of pro- versus anti-angiogenic factors, apparently distinct for physiological versus pathological angiogenesis. These studies establish novel pro angiogenic functions for AnxA1 in vascular endothelial cell sprouting, wound healing, and tumor growth and metastasis, thereby uncovering a new functional target for repairing damaged tissue and treating diseases such as cancer. They also provide critical new evidence that the tumor stroma and its microenvironment can greatly affect tumor progression and metastasis. PMID- 19805120 TI - Exploring the folding energy landscape of a series of designed consensus tetratricopeptide repeat proteins. AB - Repeat proteins contain short, tandem arrays of simple structural motifs (20-40 aa). These stack together to form nonglobular structures that are stabilized by short-range interactions from residues close in primary sequence. Unlike globular proteins, they have few, if any, long-range nonlocal stabilizing interactions. One ubiquitous repeat is the tetratricopeptide motif (TPR), a 34-aa helix-turn helix motif. In this article we describe the folding kinetics of a series of 7 designed TPR proteins that are assembled from arraying identical designed consensus repeats (CTPRan). These range from the smallest 2-repeat protein to a large 10-repeat protein (approximately 350 aa). In particular, we describe how the energy landscape changes with the addition of repeat units. The data reveal that although the CTPRa proteins have low local frustration, their highly symmetric, modular native structure is reflected in their multistate kinetics of unfolding and folding. Moreover, although the initial folding of all CTPRan proteins involves a nucleus with similar solvent accessibility, their subsequent folding to the native structure depends directly on repeat number. This corresponds to an increasingly complex landscape that culminates in CTPRa10 populating a misfolded, off-pathway intermediate. These results extend our current understanding of the malleable folding pathways of repeat proteins and highlight the consequences of adding identical repeats to the energy landscape. PMID- 19805121 TI - Continuous imaging of plasmon rulers in live cells reveals early-stage caspase-3 activation at the single-molecule level. AB - The use of plasmon coupling in metal nanoparticles has shown great potential for the optical characterization of many biological processes. Recently, we have demonstrated the use of "plasmon rulers" to observe conformational changes of single biomolecules in vitro. Plasmon rulers provide robust signals without photobleaching or blinking. Here, we show the first application of plasmon rulers to in vivo studies to observe very long trajectories of single biomolecules in live cells. We present a unique type of plasmon ruler comprised of peptide-linked gold nanoparticle satellites around a core particle, which was used as a probe to optically follow cell-signaling pathways in vivo at the single-molecule level. These "crown nanoparticle plasmon rulers" allowed us to continuously monitor trajectories of caspase-3 activity in live cells for over 2 h, providing sufficient time to observe early-stage caspase-3 activation, which was not possible by conventional ensemble analyses. PMID- 19805122 TI - Predicted poxvirus FEN1-like nuclease required for homologous recombination, double-strand break repair and full-size genome formation. AB - Poxviruses encode many if not all of the proteins required for viral genome replication in the cytoplasm of the host cell. In this context, we investigated the function of the vaccinia virus G5 protein because it belongs to the FEN1-like family of nucleases and is conserved in all poxviruses. A vaccinia virus G5 deletion mutant was severely impaired, as the yield of infectious virus was reduced by approximately two orders of magnitude. The mutant virions contained an apparently normal complement of proteins but appeared spherical rather than brick shaped and contained no detectable DNA. The inability of G5 with substitutions of the predicted catalytic aspartates to complement the deletion mutant suggested that G5 functions as a nuclease during viral DNA replication. Although the amount of viral DNA produced in the absence of G5 was similar to that made by wild-type virus, the mean size was approximately one-fourth of the genome length. Experiments with transfected plasmids showed that G5 was required for double strand break repair by homologous recombination, suggesting a similar role during vaccinia virus genome replication. PMID- 19805123 TI - NF-kappaB activity is constitutively elevated in c-Abl null fibroblasts. AB - The c-abl proto-oncogene encodes a nonreceptor tyrosine kinase involved in many cellular processes, including signaling from growth factor and antigen receptors, remodeling the cytoskeleton, and responding to DNA damage and oxidative stress. Many downstream pathways are affected by c-Abl. Elevated c-Abl kinase activity can inhibit NF-kappaB activity by stabilizing the inhibitory protein IkappaB alpha, raising the possibility that c-Abl-deficient cells might have increased NF kappaB activity. We examined the levels of NF-kappaB activity in primary mouse embryonic fibroblasts (MEFs) derived from wild-type and c-Abl knockout mice and found that the knockout MEFs indeed exhibited elevated NF-kappaB activity in response to stimulation as well as constitutively elevated NF-kappaB activity. Thus, endogenous c-Abl is a negative regulator of basal and inducible NF-kappaB activity. Examination of various points of NF-kappaB regulation revealed that unstimulated c-Abl knockout MEFs do not exhibit an increase in IkappaB alpha degradation, p65/RelA nuclear translocation, or DNA binding of NF-kappaB subunits. They do, however, show reduced levels of the histone deacetylase HDAC1, a negative regulator of basal NF-kappaB activity. Unstimulated c-Abl knockout MEFs are less responsive to induction of NF-kappaB activity by trichostatin A, an HDAC inhibitor, suggesting that c-Abl might play a role in the HDAC-mediated repression of basal NF-kappaB activity. PMID- 19805124 TI - Orai1 internalization and STIM1 clustering inhibition modulate SOCE inactivation during meiosis. AB - Store-operated Ca(2+) entry (SOCE) is a ubiquitous Ca(2+) influx pathway activated in response to depletion of intracellular Ca(2+) stores. SOCE is a primary modulator of intracellular Ca(2+) dynamics, which specify cellular responses. Interestingly, SOCE inactivates during M phase but the mechanisms involved remain unclear. SOCE is mediated by clustering of the ER Ca(2+) sensor STIM1 in response to Ca(2+) store depletion, leading to gating of the plasma membrane SOCE channel Orai1. Here we show that SOCE inactivation in meiosis is the result of internalization of Orai1 into an intracellular vesicular compartment and to the inability of STIM1 to cluster in response to store depletion. At rest, Orai1 continuously recycles between the cell membrane and an endosomal compartment. We further show that STIM1-STIM1 interactions are inhibited during meiosis, which appears to mediate the inability of STIM1 to form puncta following store depletion. In contrast, STIM1-Orai1 interactions remain functional during meiosis. Combined, the removal of Orai1 from the cell membrane and STIM1 clustering inhibition effectively uncouple store depletion from SOCE activation in meiosis. Although STIM1 is phosphorylated during meiosis, phosphomimetic and alanine substitution mutations do not modulate STIM1 clustering, arguing that phosphorylation does not mediate STIM1 clustering inhibition during meiosis. PMID- 19805125 TI - Colloquium papers: Adaptive landscapes and protein evolution. AB - The principles governing protein evolution under strong selection are important because of the recent history of evolved resistance to insecticides, antibiotics, and vaccines. One experimental approach focuses on studies of mutant proteins and all combinations of mutant sites that could possibly be intermediates in the evolutionary pathway to resistance. In organisms carrying each of the engineered proteins, a measure of protein function or a proxy for fitness is estimated. The correspondence between protein sequence and fitness is widely known as a fitness landscape or adaptive landscape. Here, we examine some empirical fitness landscapes and compare them with simulated landscapes in which the fitnesses are randomly assigned. We find that mutant sites in real proteins show significantly more additivity than those obtained from random simulations. The high degree of additivity is reflected in a summary statistic for adaptive landscapes known as the "roughness," which for the actual proteins so far examined lies in the smallest 0.5% tail of random landscapes. PMID- 19805126 TI - GABA uptake-dependent Ca(2+) signaling in developing olfactory bulb astrocytes. AB - We studied GABAergic signaling in astrocytes of olfactory bulb slices using confocal Ca(2+) imaging and two-photon Na(+) imaging. GABA evoked Ca(2+) transients in astrocytes that persisted in the presence of GABA(A) and GABA(B) receptor antagonists, but were suppressed by inhibition of GABA uptake by SNAP 5114. Withdrawal of external Ca(2+) blocked GABA-induced Ca(2+) transients, and depletion of Ca(2+) stores with cyclopiazonic acid reduced Ca(2+) transients by approximately 90%. This indicates that the Ca(2+) transients depend on external Ca(2+), but are mainly mediated by intracellular Ca(2+) release, conforming with Ca(2+)-induced Ca(2+) release. Inhibition of ryanodine receptors did not affect GABA-induced Ca(2+) transients, whereas the InsP(3) receptor blocker 2-APB inhibited the Ca(2+) transients. GABA also induced Na(+) increases in astrocytes, potentially reducing Na(+)/Ca(2+) exchange. To test whether reduction of Na(+)/Ca(2+) exchange induces Ca(2+) signaling, we inhibited Na(+)/Ca(2+) exchange with KB-R7943, which mimicked GABA-induced Ca(2+) transients. Endogenous GABA release from neurons, activated by stimulation of afferent axons or NMDA application, also triggered Ca(2+) transients in astrocytes. The significance of GABAergic Ca(2+) signaling in astrocytes for control of blood flow is demonstrated by SNAP 5114-sensitive constriction of blood vessels accompanying GABA uptake. The results suggest that GABAergic signaling is composed of GABA uptake-mediated Na(+) rises that reduce Na(+)/Ca(2+) exchange, thereby leading to a Ca(2+) increase sufficient to trigger Ca(2+)-induced Ca(2+) release via InsP(3) receptors. Hence, GABA transporters not only remove GABA from the extracellular space, but may also contribute to intracellular signaling and astrocyte function, such as control of blood flow. PMID- 19805127 TI - Direct evidence of 1,900 years of indigenous silver production in the Lake Titicaca Basin of Southern Peru. AB - Archaeological excavations at a U-shaped pyramid in the northern Lake Titicaca Basin of Peru have documented a continuous 5-m-deep stratigraphic sequence of metalworking remains. The sequence begins in the first millennium AD and ends in the Spanish Colonial period ca. AD 1600. The earliest dates associated with silver production are 1960 + or - 40 BP (2-sigma cal. 40 BC to AD 120) and 1870 + or - 40 BP (2-sigma cal. AD 60 to 240) representing the oldest known silver smelting in South America. Scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) analysis of production debris indicate a complex, multistage, high temperature technology for producing silver throughout the archaeological sequence. These data hold significant theoretical implications including the following: (i) silver production occurred before the development of the first southern Andean state of Tiwanaku, (ii) the location and process of silverworking remained consistent for 1,500 years even though political control of the area cycled between expansionist states and smaller chiefly polities, and (iii) that U-shaped structures were the location of ceremonial, residential, and industrial activities. PMID- 19805128 TI - Dissociation of circadian and light inhibition of melatonin release through forced desynchronization in the rat. AB - Pineal melatonin release exhibits a circadian rhythm with a tight nocturnal pattern. Melatonin synthesis is regulated by the master circadian clock within the hypothalamic suprachiasmatic nucleus (SCN) and is also directly inhibited by light. The SCN is necessary for both circadian regulation and light inhibition of melatonin synthesis and thus it has been difficult to isolate these two regulatory limbs to define the output pathways by which the SCN conveys circadian and light phase information to the pineal. A 22-h light-dark (LD) cycle forced desynchrony protocol leads to the stable dissociation of rhythmic clock gene expression within the ventrolateral SCN (vlSCN) and the dorsomedial SCN (dmSCN). In the present study, we have used this protocol to assess the pattern of melatonin release under forced desynchronization of these SCN subregions. In light of our reported patterns of clock gene expression in the forced desynchronized rat, we propose that the vlSCN oscillator entrains to the 22-h LD cycle whereas the dmSCN shows relative coordination to the light-entrained vlSCN, and that this dual-oscillator configuration accounts for the pattern of melatonin release. We present a simple mathematical model in which the relative coordination of a single oscillator within the dmSCN to a single light-entrained oscillator within the vlSCN faithfully portrays the circadian phase, duration and amplitude of melatonin release under forced desynchronization. Our results underscore the importance of the SCN's subregional organization to both photic input processing and rhythmic output control. PMID- 19805129 TI - Toggle involving cis-interfering noncoding RNAs controls variegated gene expression in yeast. AB - The identification of specific functional roles for the numerous long noncoding (nc)RNAs found in eukaryotic transcriptomes is currently a matter of intense study amid speculation that these ncRNAs have key regulatory roles. We have identified a pair of cis-interfering ncRNAs in yeast that contribute to the control of variegated gene expression at the FLO11 locus by implementing a regulatory circuit that toggles between two stable states. These capped, polyadenylated ncRNAs are transcribed across the large intergenic region upstream of the FLO11 ORF. As with mammalian long intervening (li)ncRNAs, these yeast ncRNAs (ICR1 and PWR1) are themselves regulated by transcription factors (Sfl1 and Flo8) and chromatin remodelers (Rpd3L) that are key elements in phenotypic transitions in yeast. The mechanism that we describe explains the unanticipated role of a histone deacetylase complex in activating gene expression, because Rpd3L mutants force the ncRNA circuit into a state that silences the expression of the adjacent variegating gene. PMID- 19805130 TI - Insulin resistance is a cellular antioxidant defense mechanism. AB - We know a great deal about the cellular response to starvation via AMPK, but less is known about the reaction to nutrient excess. Insulin resistance may be an appropriate response to nutrient excess, but the cellular sensors that link these parameters remain poorly defined. In the present study we provide evidence that mitochondrial superoxide production is a common feature of many different models of insulin resistance in adipocytes, myotubes, and mice. In particular, insulin resistance was rapidly reversible upon exposure to agents that act as mitochondrial uncouplers, ETC inhibitors, or mitochondrial superoxide dismutase (MnSOD) mimetics. Similar effects were observed with overexpression of mitochondrial MnSOD. Furthermore, acute induction of mitochondrial superoxide production using the complex III antagonist antimycin A caused rapid attenuation of insulin action independently of changes in the canonical PI3K/Akt pathway. These results were validated in vivo in that MnSOD transgenic mice were partially protected against HFD induced insulin resistance and MnSOD+/- mice were glucose intolerant on a standard chow diet. These data place mitochondrial superoxide at the nexus between intracellular metabolism and the control of insulin action potentially defining this as a metabolic sensor of energy excess. PMID- 19805131 TI - Quantum-mechanics-derived 13Calpha chemical shift server (CheShift) for protein structure validation. AB - A server (CheShift) has been developed to predict (13)C(alpha) chemical shifts of protein structures. It is based on the generation of 696,916 conformations as a function of the phi, psi, omega, chi1 and chi2 torsional angles for all 20 naturally occurring amino acids. Their (13)C(alpha) chemical shifts were computed at the DFT level of theory with a small basis set and extrapolated, with an empirically-determined linear regression formula, to reproduce the values obtained with a larger basis set. Analysis of the accuracy and sensitivity of the CheShift predictions, in terms of both the correlation coefficient R and the conformational-averaged rmsd between the observed and predicted (13)C(alpha) chemical shifts, was carried out for 3 sets of conformations: (i) 36 x-ray derived protein structures solved at 2.3 A or better resolution, for which sets of (13)C(alpha) chemical shifts were available; (ii) 15 pairs of x-ray and NMR derived sets of protein conformations; and (iii) a set of decoys for 3 proteins showing an rmsd with respect to the x-ray structure from which they were derived of up to 3 A. Comparative analysis carried out with 4 popular servers, namely SHIFTS, SHIFTX, SPARTA, and PROSHIFT, for these 3 sets of conformations demonstrated that CheShift is the most sensitive server with which to detect subtle differences between protein models and, hence, to validate protein structures determined by either x-ray or NMR methods, if the observed (13)C(alpha) chemical shifts are available. CheShift is available as a web server. PMID- 19805132 TI - Common variants on chromosome 2 and risk of primary open-angle glaucoma in the Afro-Caribbean population of Barbados. AB - Primary open-angle glaucoma (POAG) is the second leading cause of blindness worldwide. Although a number of genetic loci have shown association or genetic linkage to monogenic forms of POAG, the identified genes and loci do not appear to have a major role in the common POAG phenotype. We seek to identify genetic loci that appear to be major risk factors for POAG in the Afro-Caribbean population of Barbados, West Indies. We performed linkage analyses in 146 multiplex families ascertained through the Barbados Family Study of Glaucoma (BFSG) and identified a strong linkage signal on chromosome 2p (logarithm of odds score = 6.64 at = 0 with marker D2S2156). We subsequently performed case-control analyses using unrelated affected individuals and unaffected controls. A set of SNPs on chromosome 2p was evaluated in two independent groups of BFSG participants, a discovery group (130 POAG cases, 65 controls) and a replication group (122 POAG cases, 65 controls), and a strong association was identified with POAG and rs12994401 in both groups (P < 3.34 E-09 and P < 1.21E-12, respectively). The associated SNPs form a common disease haplotype. In summary, we have identified a locus with a major impact on susceptibility to the common POAG phenotype in an Afro-Caribbean population in Barbados. Our approach illustrates the merit of using an isolated population enriched with common disease variants as an efficient method to identify genetic underpinning of POAG. PMID- 19805133 TI - A CREB-C/EBPbeta cascade induces M2 macrophage-specific gene expression and promotes muscle injury repair. AB - Macrophages play an essential role in the resolution of tissue damage through removal of necrotic cells, thus paving the way for tissue regeneration. Macrophages also directly support the formation of new tissue to replace the injury, through their acquisition of an anti-inflammatory, or M2, phenotype, characterized by a gene expression program that includes IL-10, the IL-13 receptor, and arginase 1. We report that deletion of two CREB-binding sites from the Cebpb promoter abrogates Cebpb induction upon macrophage activation. This blocks the downstream induction of M2-specific Msr1, Il10, II13ra, and Arg-1 genes, whereas the inflammatory (M1) genes Il1, Il6, Tnfa, and Il12 are not affected. Mice carrying the mutated Cebpb promoter (betaDeltaCre) remove necrotic tissue from injured muscle, but exhibit severe defects in muscle fiber regeneration. Conditional deletion of the Cebpb gene in muscle cells does not affect regeneration, showing that the C/EBPbeta cascade leading to muscle repair is muscle-extrinsic. While betaDeltaCre macrophages efficiently infiltrate injured muscle they fail to upregulate Cebpb, leading to decreased Arg-1 expression. CREB-mediated induction of Cebpb expression is therefore required in infiltrating macrophages for upregulation of M2-specific genes and muscle regeneration, providing a direct genetic link between these two processes. PMID- 19805134 TI - Repeated climate-linked host shifts have promoted diversification in a temperate clade of leaf-mining flies. AB - A central but little-tested prediction of "escape and radiation" coevolution is that colonization of novel, chemically defended host plant clades accelerates insect herbivore diversification. That theory, in turn, exemplifies one side of a broader debate about the relative influence on clade dynamics of intrinsic (biotic) vs. extrinsic (physical-environmental) forces. Here, we use a fossil calibrated molecular chronogram to compare the effects of a major biotic factor (repeated shift to a chemically divergent host plant clade) and a major abiotic factor (global climate change) on the macroevolutionary dynamics of a large Cenozoic radiation of phytophagous insects, the leaf-mining fly genus Phytomyza (Diptera: Agromyzidae). We find one of the first statistically supported examples of consistently elevated net diversification accompanying shift to new plant clades. In contrast, we detect no significant direct effect on diversification of major global climate events in the early and late Oligocene. The broader paleoclimatic context strongly suggests, however, that climate change has at times had a strong indirect influence through its effect on the biotic environment. Repeated rapid Miocene radiation of these flies on temperate herbaceous asterids closely corresponds to the dramatic, climate-driven expansion of seasonal, open habitats. PMID- 19805135 TI - An unexpected role for TASK-3 potassium channels in network oscillations with implications for sleep mechanisms and anesthetic action. AB - TASK channels are acid-sensitive and anesthetic-activated members of the family of two-pore-domain potassium channels. We have made the surprising discovery that the genetic ablation of TASK-3 channels eliminates a specific type of theta oscillation in the cortical electroencephalogram (EEG) resembling type II theta (4-9 Hz), which is thought to be important in processing sensory stimuli before initiating motor activity. In contrast, ablation of TASK-1 channels has no effect on theta oscillations. Despite the absence of type II theta oscillations in the TASK-3 knockout (KO) mice, the related type I theta, which has certain neuronal pathways in common and is involved in exploratory behavior, is unaffected. In addition to the absence of type II theta oscillations, the TASK-3 KO animals show marked alterations in both anesthetic sensitivity and natural sleep behavior. Their sensitivity to halothane, a potent activator of TASK channels, is greatly reduced, whereas their sensitivity to cyclopropane, which does not activate TASK 3 channels, is unchanged. The TASK-3 KO animals exhibit a slower progression from their waking to sleeping states and, during their sleeping period, their sleep episodes as well as their REM theta oscillations are more fragmented. These results imply a previously unexpected role for TASK-3 channels in the cellular mechanisms underlying these behaviors and suggest that endogenous modulators of these channels may regulate theta oscillations. PMID- 19805136 TI - The conserved Est1 protein stimulates telomerase DNA extension activity. AB - The first telomerase cofactor identified was the budding yeast protein Est1, which is conserved through humans. While it is evident that Est1 is required for telomere DNA maintenance, understanding its mechanistic contributions to telomerase regulation has been limited. In vitro, the primary effect of Est1 is to activate telomerase-mediated DNA extension. Although Est1 displayed specific DNA and RNA binding, neither activity contributed significantly to telomerase stimulation. Rather Est1 mediated telomerase upregulation through direct contacts with the reverse transcriptase subunit. In addition to intrinsic Est1 functions, we found that Est1 cooperatively activated telomerase in conjunction with Cdc13 and that the combinatorial effect was dependent upon a known salt-bridge interaction between Est1 (K444) and Cdc13 (E252). Our studies provide insights into the molecular events used to control the enzymatic activity of the telomerase holoenzyme. PMID- 19805137 TI - DNA polymerase epsilon and delta proofreading suppress discrete mutator and cancer phenotypes in mice. AB - Organisms require faithful DNA replication to avoid deleterious mutations. In yeast, replicative leading- and lagging-strand DNA polymerases (Pols epsilon and delta, respectively) have intrinsic proofreading exonucleases that cooperate with each other and mismatch repair to limit spontaneous mutation to less than 1 per genome per cell division. The relationship of these pathways in mammals and their functions in vivo are unknown. Here we show that mouse Pol epsilon and delta proofreading suppress discrete mutator and cancer phenotypes. We found that inactivation of Pol epsilon proofreading elevates base-substitution mutations and accelerates a unique spectrum of spontaneous cancers; the types of tumors are entirely different from those triggered by loss of Pol delta proofreading. Intercrosses of Pol epsilon-, Pol delta-, and mismatch repair-mutant mice show that Pol epsilon and delta proofreading act in parallel pathways to prevent spontaneous mutation and cancer. These findings distinguish Pol epsilon and delta functions in vivo and reveal tissue-specific requirements for DNA replication fidelity. PMID- 19805138 TI - Structural relationships among proteins with different global topologies and their implications for function annotation strategies. AB - It has become increasingly apparent that geometric relationships often exist between regions of two proteins that have quite different global topologies or folds. In this article, we examine whether such relationships can be used to infer a functional connection between the two proteins in question. We find, by considering a number of examples involving metal and cation binding, sugar binding, and aromatic group binding, that geometrically similar protein fragments can share related functions, even if they have been classified as belonging to different folds and topologies. Thus, the use of classifications inevitably limits the number of functional inferences that can be obtained from the comparative analysis of protein structures. In contrast, the development of interactive computational tools that recognize the "continuous" nature of protein structure/function space, by increasing the number of potentially meaningful relationships that are considered, may offer a dramatic enhancement in the ability to extract information from protein structure databases. We introduce the MarkUs server, that embodies this strategy and that is designed for a user interested in developing and validating specific functional hypotheses. PMID- 19805139 TI - Retinoid-related orphan nuclear receptor RORbeta is an early-acting factor in rod photoreceptor development. AB - Rods and cones are morphologically and developmentally distinct photoreceptor types with different functions in vision. Cones mediate daylight and color vision and in most mammals express M and S opsin photopigments for sensitivity to medium long and short light wavelengths, respectively. Rods mediate dim light vision and express rhodopsin photopigment. The transcription factor networks that direct differentiation of each photoreceptor type are incompletely defined. Here, we report that Rorb(-/-) mice lacking retinoid-related orphan nuclear receptor beta lose rods but overproduce primitive S cones that lack outer segments. The phenotype reflects pronounced plasticity between rod and cone lineages and resembles that described for Nrl(-/-) mice lacking neural retina leucine zipper factor. Rorb(-/-) mice lack Nrl expression and reexpression of Nrl in Rorb(-/-) mice converts cones to rod-like cells. Thus, Rorb directs rod development and does so at least in part by inducing the Nrl-mediated pathway of rod differentiation. PMID- 19805140 TI - Control of bud activation by an auxin transport switch. AB - In many plant species only a small proportion of buds yield branches. Both the timing and extent of bud activation are tightly regulated to produce specific branching architectures. For example, the primary shoot apex can inhibit the activation of lateral buds. This process is termed apical dominance and is dependent on the plant hormone auxin moving down the main stem in the polar auxin transport stream. We use a computational model and mathematical analysis to show that apical dominance can be explained in terms of an auxin transport switch established by the temporal precedence between competing auxin sources. Our model suggests a mechanistic basis for the indirect action of auxin in bud inhibition and captures the effects of diverse genetic and physiological manipulations. In particular, the model explains the surprising observation that highly branched Arabidopsis phenotypes can exhibit either high or low auxin transport. PMID- 19805142 TI - A robust method for searching the smallest set of smallest rings with a path included distance matrix. AB - The perception of rings in graphs is widely used in many fields of science and engineering. Algorithms developed in the chemistry community, called smallest set of smallest rings (SSSR), applicable only for simple graphs or chemical structures. In contrast, algorithms developed by the computer science community, called minimum cycle basis (MCB) are identical to SSSR yet exhibit greater robustness. MCB-based algorithms can correctly reveal all rings in any complex graph. However, they are slow when applied to large complex graphs due to the inherent limitations of the algorithms used. Here, we suggest a heuristic method called RP-Path. This method is a robust, simple, and fast search method with O(n(3)) runtime algorithm that correctly identifies the SSSR of all of the test case of complex graphs by using approach different from the MCB-based method. Both the robustness and improvement in speed are achieved by using a path included distance matrix and describing the characteristic features of rings in the matrix. This method is accurate and faster than any other methods and may find many application in various fields of science and engineering that use complicated graphs with thousands of nodes. PMID- 19805141 TI - Adoptively transferred effector cells derived from naive rather than central memory CD8+ T cells mediate superior antitumor immunity. AB - Effector cells derived from central memory CD8(+) T cells were reported to engraft and survive better than those derived from effector memory populations, suggesting that they are superior for use in adoptive immunotherapy studies. However, previous studies did not evaluate the relative efficacy of effector cells derived from naive T cells. We sought to investigate the efficacy of tumor specific effector cells derived from naive or central memory T-cell subsets using transgenic or retrovirally transduced T cells engineered to express a tumor specific T-cell receptor. We found that naive, rather than central memory T cells, gave rise to an effector population that mediated superior antitumor immunity upon adoptive transfer. Effector cells developed from naive T cells lost the expression of CD62L more rapidly than those derived from central memory T cells, but did not acquire the expression of KLRG-1, a marker for terminal differentiation and replicative senescence. Consistent with this KLRG-1(-) phenotype, naive-derived cells were capable of a greater proliferative burst and had enhanced cytokine production after adoptive transfer. These results indicate that insertion of genes that confer antitumor specificity into naive rather than central memory CD8(+) T cells may allow superior efficacy upon adoptive transfer. PMID- 19805143 TI - aP2-Cre-mediated inactivation of acetyl-CoA carboxylase 1 causes growth retardation and reduced lipid accumulation in adipose tissues. AB - Adipose tissue is one of the major sites for fatty acid synthesis and lipid storage. We generated adipose (fat)-specific ACC1 knockout (FACC1KO) mice using the aP2-Cre/loxP system. FACC1KO mice showed prenatal growth retardation; after weaning, however, their weight gain was comparable to that of wild-type (WT) mice on a normal diet. Under lipogenic conditions of fasting/re-feeding a fat-free diet, lipid accumulation in adipose tissues of FACC1KO mice was significantly decreased; this is consistent with a 50-66% reduction in the ACC activity in these tissues compared with that of WT mice. Surprisingly, FACC1KO mice manifested skeletal growth retardation phenotype accompanied by decreased chondrocyte proliferation in the growth plate and lower trabecular bone density. In addition, there was about a 30% decrease in serum insulin-like growth factor I (IGF1), and while the serum leptin level was decreased by about 50%, it did not counteract the osteopenic effects of IGF1 on the bone. Fatty acid analyses of mutant bone lipids revealed relatively higher levels of C18:2n-6 and C18:3n-3 and lower levels of their elongation C20 homologs than that of WT cohorts, leading to lower levels of C20 homologs and bone development. Moreover, aP2-Cre-mediated ACC1 inactivation in bone tissue led to a decreased number of osteoblasts but not of osteoclasts. The downregulation of ACC1 on osteoblastogenesis may be the cause for the osteopenia phenotype of FACC1KO bone homeostasis. PMID- 19805144 TI - AnkyrinG is required to maintain axo-dendritic polarity in vivo. AB - Neurons are highly polarized cells that extend a single axon and several dendrites. Studies with cultured neurons indicate that the proximal portion of the axon, denoted as the axon initial segment (AIS), maintains neuronal polarity in vitro. The membrane-adaptor protein ankyrinG (ankG) is an essential component of the AIS. To determine the relevance of ankG for neuronal polarity in vivo, we studied mice with a cerebellum-specific ankG deficiency. Strikingly, ankG depleted axons develop protrusions closely resembling dendritic spines. Such axonal spines are enriched with postsynaptic proteins, including ProSAP1/Shank2 and ionotropic and metabotropic glutamate receptors. In addition, immunofluorescence indicated that axonal spines are contacted by presynaptic glutamatergic boutons. For further analysis, double mutants were obtained by crossbreeding ankG(-/-) mice with L7/Purkinje cell-specific promoter 2 (PCP2) mice expressing enhanced green fluorescent protein (EGFP) in Purkinje cells (PCs). This approach allowed precise confocal microscopic mapping of EGFP positive spiny axons and their subsequent identification at the electron microscopic level. Ultrastructurally, axonal spines contained a typical postsynaptic density and established asymmetric excitatory synapses with presynaptic boutons containing synaptic vesicles. In the shaft of spiny axons, typical ultrastructural features of the AIS, including the membrane-associated dense undercoating and cytoplasmic bundles of microtubules, were absent. Finally, using time-lapse imaging of organotypic cerebellar slice cultures, we demonstrate that nonspiny PC axons of EGFP-positive/ankG(-/-) mice acquire a spiny phenotype within a time range of only 3 days. Collectively, these findings demonstrate that axons of ankG-deficient mice acquire hallmark features of dendrites. AnkG thus is important for maintaining appropriate axo-dendritic polarity in vivo. PMID- 19805145 TI - E3 ubiquitin ligase COP1 regulates the stability and functions of MTA1. AB - Metastasis-associated protein 1 (MTA1), a component of the nucleosome remodeling and histone deacetylation (NuRD) complex, is widely upregulated in human cancers. However, the mechanism for regulating its protein stability remains unknown. Here we report that MTA1 is an ubiquitinated protein and targeted by the RING-finger E3 ubiquitin-protein ligase constitutive photomorphogenesis protein 1 (COP1) for degradation via the ubiquitin-proteasome pathway. Induced expression of wild-type COP1 but not its RING motif mutants promotes the ubiquitination and degradation of MTA1, indicating that the ligase activity is required for the COP1-mediated proteolysis of MTA1. Conversely, depletion of endogenous COP1 resulted in a marked decrease in MTA1 ubiquitination, accompanied by a pronounced accumulation of MTA1 protein. MTA1, in turn, destabilizes COP1 by promoting its autoubiquitination, thus creating a tight feedback loop that regulates both MTA1 and COP1 protein stability. Accordingly, disruption of the COP1-mediated proteolysis by ionizing radiation leads to MTA1 stabilization, accompanied by an increased coregulatory function of MTA1 on its target. Furthermore, we discovered that MTA1 is required for optimum DNA double-strand break repair after ionizing radiation. These findings provide novel insights into the regulation of MTA1 protein and reveal a novel function of MTA1 in DNA damage response. PMID- 19805146 TI - Fungal infection counters insecticide resistance in African malaria mosquitoes. AB - The evolution of insecticide resistance in mosquitoes is threatening the effectiveness and sustainability of malaria control programs in various parts of the world. Through their unique mode of action, entomopathogenic fungi provide promising alternatives to chemical control. However, potential interactions between fungal infection and insecticide resistance, such as cross-resistance, have not been investigated. We show that insecticide-resistant Anopheles mosquitoes remain susceptible to infection with the fungus Beauveria bassiana. Four different mosquito strains with high resistance levels against pyrethroids, organochlorines, or carbamates were equally susceptible to B. bassiana infection as their baseline counterparts, showing significantly reduced mosquito survival. Moreover, fungal infection reduced the expression of resistance to the key public health insecticides permethrin and dichlorodiphenyltrichloroethane. Mosquitoes preinfected with B. bassiana or Metarhizium anisopliae showed a significant increase in mortality after insecticide exposure compared with uninfected control mosquitoes. Our results show a high potential utility of fungal biopesticides for complementing existing vector control measures and provide products for use in resistance management strategies. PMID- 19805147 TI - Implicit sampling for particle filters. AB - We present a particle-based nonlinear filtering scheme, related to recent work on chainless Monte Carlo, designed to focus particle paths sharply so that fewer particles are required. The main features of the scheme are a representation of each new probability density function by means of a set of functions of Gaussian variables (a distinct function for each particle and step) and a resampling based on normalization factors and Jacobians. The construction is demonstrated on a standard, ill-conditioned test problem. PMID- 19805148 TI - Inhibition of protein kinase C signaling protects prefrontal cortex dendritic spines and cognition from the effects of chronic stress. AB - The prefrontal cortex r regulates behavior, cognition, and emotion by using working memory. Prefrontal functions are impaired by stress exposure. Acute, stress-induced deficits arise from excessive protein kinase C (PKC) signaling, which diminishes prefrontal neuronal firing. Chronic stress additionally produces architectural changes, reducing dendritic complexity and spine density of cortico cortical pyramidal neurons, thereby disrupting excitatory working memory networks. In vitro studies have found that sustained PKC activity leads to spine loss from hippocampal-cultured neurons, suggesting that PKC may contribute to spine loss during chronic stress exposure. The present study tested whether inhibition of PKC with chelerythrine before daily stress would protect prefrontal spines and working memory. We found that inhibition of PKC rescued working memory impairments and reversed distal apical dendritic spine loss in layer II/III pyramidal neurons of rat prelimbic cortex. Greater spine density predicted better cognitive performance, the first direct correlation between pyramidal cell structure and working memory abilities. These findings suggest that PKC inhibitors may be neuroprotective in disorders with dysregulated PKC signaling such as bipolar disorder, schizophrenia, post-traumatic stress disorder, and lead poisoning--conditions characterized by impoverished prefrontal structural and functional integrity. PMID- 19805149 TI - Interareal coordination of columnar architectures during visual cortical development. AB - The formation of cortical columns is often conceptualized as a local process in which synaptic microcircuits confined to the volume of the emerging column are established and selectively refined. Many neurons, however, while wiring up locally are simultaneously building macroscopic circuits spanning widely distributed brain regions, such as different cortical areas or the two brain hemispheres. Thus, it is conceivable that interareal interactions shape the local column layout. Here we show that the columnar architectures of different areas of the cat visual cortex in fact develop in a coordinated manner, not adequately described as a local process. This is revealed by comparing the layouts of orientation columns (i) in left/right pairs of brain hemispheres and (ii) in areas V1 and V2 of individual brain hemispheres. Whereas the size of columns varied strongly within all areas considered, columns in different areas were typically closely matched in size if they were mutually connected. During development, we find that such mutually connected columns progressively become better matched in size as the late phase of the critical period unfolds. Our results suggest that one function of critical-period plasticity is to progressively coordinate the functional architectures of different cortical areas -even across hemispheres. PMID- 19805150 TI - Identification of tumor-initiating cells in a highly aggressive brain tumor using promoter activity of nucleostemin. AB - Controversy remains over whether the cancer stem cell (CSC) theory applies to all tumors. To determine whether cells within a highly aggressive solid tumor are stochastically or hierarchically organized, we combined a reporter system where the nucleostemin (NS) promoter drives GFP expression (termed NS-GFP) with a mouse brain tumor model induced by retroviral Ras expression on a p16(Ink4a)/p19(Arf) deficient background. The NS-GFP system allowed us to monitor the differentiation process of normal neural stem/precursor cells by analyzing GFP fluorescence intensity. In tumor-bearing mice, despite the very high frequency of tumorigenic cells, we successfully identified the NS-GFP(+) cells as tumor-initiating cells (T-ICs). The clonal studies conclusively established that phenotypical heterogeneity can exist among the cells comprising a genetically homogeneous tumor, suggesting that this aggressive brain tumor follows the CSC model. Detailed analyses of the NS-GFP(+) brain tumor cells revealed that T-ICs showed activation of the receptor tyrosine kinase c-Met, which functions in tumor invasiveness. Thus, the NS-GFP system provides a powerful tool to elucidate stem cell biology in normal and malignant tissues. PMID- 19805151 TI - MEIG1 is essential for spermiogenesis in mice. AB - Spermatogenesis can be divided into three stages: spermatogonial mitosis, meiosis of spermatocytes, and spermiogenesis. During spermiogenesis, spermatids undergo dramatic morphological changes including formation of a flagellum and chromosomal packaging and condensation of the nucleus into the sperm head. The genes regulating the latter processes are largely unknown. We previously discovered that a bi-functional gene, Spag16, is essential for spermatogenesis. SPAG16S, the 35 kDa, testis-specific isoform derived from the Spag16 gene, was found to bind to meiosis expressed gene 1 product (MEIG1), a protein originally thought to play a role in meiosis. We inactivated the Meig1 gene and, unexpectedly, found that Meig1 mutant male mice had no obvious defect in meiosis, but were sterile as a result of impaired spermatogenesis at the stage of elongation and condensation. Transmission electron microscopy revealed that the manchette, a microtubular organelle essential for sperm head and flagellar formation was disrupted in spermatids of MEIG1-deficient mice. We also found that MEIG1 associates with the Parkin co-regulated gene (PACRG) protein, and that testicular PACRG protein is reduced in MEIG1-deficient mice. PACRG is thought to play a key role in assembly of the axonemes/flagella and the reproductive phenotype of Pacrg-deficient mice mirrors that of the Meig1 mutant mice. Our findings reveal a critical role for the MEIG1/PARCG partnership in manchette structure and function and the control of spermiogenesis. PMID- 19805152 TI - Comparative genomic analysis of Lactobacillus rhamnosus GG reveals pili containing a human- mucus binding protein. AB - To unravel the biological function of the widely used probiotic bacterium Lactobacillus rhamnosus GG, we compared its 3.0-Mbp genome sequence with the similarly sized genome of L. rhamnosus LC705, an adjunct starter culture exhibiting reduced binding to mucus. Both genomes demonstrated high sequence identity and synteny. However, for both strains, genomic islands, 5 in GG and 4 in LC705, punctuated the colinearity. A significant number of strain-specific genes were predicted in these islands (80 in GG and 72 in LC705). The GG-specific islands included genes coding for bacteriophage components, sugar metabolism and transport, and exopolysaccharide biosynthesis. One island only found in L. rhamnosus GG contained genes for 3 secreted LPXTG-like pilins (spaCBA) and a pilin-dedicated sortase. Using anti-SpaC antibodies, the physical presence of cell wall-bound pili was confirmed by immunoblotting. Immunogold electron microscopy showed that the SpaC pilin is located at the pilus tip but also sporadically throughout the structure. Moreover, the adherence of strain GG to human intestinal mucus was blocked by SpaC antiserum and abolished in a mutant carrying an inactivated spaC gene. Similarly, binding to mucus was demonstrated for the purified SpaC protein. We conclude that the presence of SpaC is essential for the mucus interaction of L. rhamnosus GG and likely explains its ability to persist in the human intestinal tract longer than LC705 during an intervention trial. The presence of mucus-binding pili on the surface of a nonpathogenic Gram positive bacterial strain reveals a previously undescribed mechanism for the interaction of selected probiotic lactobacilli with host tissues. PMID- 19805153 TI - Human gut microbiome adopts an alternative state following small bowel transplantation. AB - Small bowel transplants provide an exceptional opportunity for long-term study of the microbial ecology of the human small bowel. The ileostomy created at time of transplant for ongoing monitoring of the allograft provides access to samples of ileal effluent and mucosal biopsies. In this study, we used qPCR to assay the bacterial population of the small bowel lumen of 17 small bowel transplant patients over time. Surprisingly, the posttransplant microbial community was found to be dominated by Lactobacilli and Enterobacteria, both typically facultative anaerobes. This represents an inversion of the normal community that is dominated instead by the strictly anaerobic Bacteroides and Clostridia. We found this inverted community also in patients with ileostomies who did not receive a transplant, suggesting that the ileostomy itself is the primary ecological determinant shaping the microbiota. After surgical closure of the ileostomy, the community reverted to the normal structure. Therefore, we hypothesized that the ileostomy allows oxygen into the otherwise anaerobic distal ileum, thus driving the transition from one microbial community structure to another. Supporting this hypothesis, metabolomic profiling of both communities demonstrated an enrichment for metabolites associated with aerobic respiration in samples from patients with open ileostomies. Viewed from an ecological perspective, the two communities constitute alternative stable states of the human ileum. That the small bowel appears to function normally despite these dramatic shifts suggests that its ecological resilience is greater than previously realized. PMID- 19805154 TI - Efficient IgM assembly and secretion require the plasma cell induced endoplasmic reticulum protein pERp1. AB - Plasma cells daily secrete their own mass in antibodies, which fold and assemble in the endoplasmic reticulum (ER). To reach these levels, cells require pERp1, a novel lymphocyte-specific small ER-resident protein, which attains expression levels as high as BiP when B cells differentiate into plasma cells. Although pERp1 has no homology with known ER proteins, it does contain a CXXC motif typical for oxidoreductases. In steady state, the CXXC cysteines are locked by two parallel disulfide bonds with a downstream C(X)(6)C motif, and pERp1 displays only modest oxidoreductase activity. pERp1 emerged as a dedicated folding factor for IgM, associating with both heavy and light chains and promoting assembly and secretion of mature IgM. PMID- 19805155 TI - TRAF5 is a critical mediator of in vitro signals and in vivo functions of LMP1, the viral oncogenic mimic of CD40. AB - The cytoplasmic signaling protein TNF receptor-associated factor 5 (TRAF5) has been implicated in several biological roles in T-lymphocyte responses. However, a clear connection between in vivo TRAF5 immune cell functions and specific signaling pathways has not been made. This study shows that TRAF5 associated strongly with the viral oncogenic CD40 mimic latent membrane protein 1 (LMP1), in contrast to weaker association with CD40, for which it has been shown to play a modest role. LMP1 uses specific TRAFs differently than CD40, resulting in amplified and dysregulated CD40-like activation of B lymphocytes. When the cytoplasmic domain of LMP1 is expressed as a transgenic replacement for CD40 in mouse B cells, the resulting mouse exhibits measures of B-cell hyperactivity such as splenomegaly, lymphadenopathy, elevated serum IL-6, elevated serum autoantibodies, and abnormal splenic architecture. Thus, in contrast to CD40, TRAF5 may have an important nonredundant role as a positive mediator of LMP1 signaling and functions in B cells. To test this hypothesis, mice were created that express mCD40LMP1 in place of CD40, and are either sufficient or deficient in TRAF5. Results revealed that TRAF5 plays a critical role in LMP1-mediated c Jun kinase signaling and is required for much of the abnormal phenotype observed in mCD40LMP1 transgenic mice. This is the first report showing a major requirement for TRAF5 in signaling by a specific receptor both in vitro and in vivo, as well as playing an important role in biological function in B lymphocytes. PMID- 19805156 TI - Triplet repeat length bias and variation in the human transcriptome. AB - Length variation in short tandem repeats (STRs) is an important family of DNA polymorphisms with numerous applications in genetics, medicine, forensics, and evolutionary analysis. Several major diseases have been associated with length variation of trinucleotide (triplet) repeats including Huntington's disease, hereditary ataxias and spinobulbar muscular atrophy. Using the reference human genome, we have catalogued all triplet repeats in genic regions. This data revealed a bias in noncoding DNA repeat lengths. It also enabled a survey of repeat-length polymorphisms (RLPs) in human genomes and a comparison of the rate of polymorphism in humans versus divergence from chimpanzee. For short repeats, this analysis of three human genomes reveals a relatively low RLP rate in exons and, somewhat surprisingly, in introns. All short RLPs observed in multiple genomes are biallelic (at least in this small sample). In contrast, long repeats are highly polymorphic and some long RLPs are multiallelic. For long repeats, the chimpanzee sequence frequently differs from all observed human alleles. This suggests a high expansion/contraction rate in all long repeats. Expansions and contractions are not, however, affected by natural selection discernable from our comparison of human-chimpanzee divergence with human RLPs. Our catalog of human triplet repeats and their surrounding flanking regions can be used to produce a cost-effective whole-genome assay to test individuals. This repeat assay could someday complement SNP arrays for producing tests that assess the risk of an individual to develop a disease, or become part of personalized genomic strategy that provides therapeutic guidance with respect to drug response. PMID- 19805157 TI - pERp1 is significantly up-regulated during plasma cell differentiation and contributes to the oxidative folding of immunoglobulin. AB - Plasma cells can synthesize and secrete thousands of Ig molecules per second, which are folded and assembled in the endoplasmic reticulum (ER) and are likely to place unusually high demands on the resident chaperones and folding enzymes. We have discovered a new resident ER protein (pERp1) that is a component of the BiP chaperone complex. PERp1 is substantially up-regulated during B to plasma cell differentiation and can be induced in B cell lines by some UPR activators, arguing that it represents a potentially new class of conditional UPR targets. In LPS-stimulated murine splenocytes, pERp1 interacted covalently via a disulfide bond with IgM monomers and noncovalently with other Ig assembly intermediates. Knockdown and overexpression experiments revealed that pERp1 promoted correct oxidative folding of Ig heavy chains and prevented off-pathway assembly intermediates. Although pERp1 has no homology with known chaperones or folding enzymes, it possesses a thioredoxin-like active site motif (CXXC), which is the signature of oxidoreductases. Mutation of this sequence did not affect its in vivo activity, suggesting that pERp1 is either a unique type of oxidoreductase or a previously unidentified class of molecular chaperone that is dedicated to enhancing the oxidative folding of Ig precursors. PMID- 19805159 TI - Retraction. PKNOX2 gene is significantly associated with substance dependence in European-origin women. PMID- 19805158 TI - Clonality of mouse and human cardiomyogenesis in vivo. AB - An analysis of the clonality of cardiac progenitor cells (CPCs) and myocyte turnover in vivo requires genetic tagging of the undifferentiated cells so that the clonal marker of individual mother cells is traced in the specialized progeny. CPC niches in the atria and apex of the mouse heart were infected with a lentivirus carrying EGFP, and the destiny of the tagged cells was determined 1-5 months later. A common integration site was identified in isolated CPCs, cardiomyocytes, endothelial cells (ECs), and fibroblasts, documenting CPC self renewal and multipotentiality and the clonal origin of the differentiated cell populations. Subsequently, the degree of EGFP-lentiviral infection of CPCs was evaluated 2-4 days after injection, and the number of myocytes expressing the reporter gene was measured 6 months later. A BrdU pulse-chasing protocol was also introduced as an additional assay for the analysis of myocyte turnover. Over a period of 6 months, each EGFP-positive CPC divided approximately eight times generating 230 cardiomyocytes; this value was consistent with the number of newly formed cells labeled by BrdU. To determine whether, human CPCs (hCPCs) are self renewing and multipotent, these cells were transduced with the EGFP-lentivirus and injected after acute myocardial infarction in immunosuppressed rats. hCPCs, myocytes, ECs, and fibroblasts collected from the regenerated myocardium showed common viral integration sites in the human genome. Thus, our results indicate that the adult heart contains a pool of resident stem cells that regulate cardiac homeostasis and repair. PMID- 19805160 TI - Evidence for adaptive radiation from a phylogenetic study of plant defenses. AB - One signature of adaptive radiation is a high level of trait change early during the diversification process and a plateau toward the end of the radiation. Although the study of the tempo of evolution has historically been the domain of paleontologists, recently developed phylogenetic tools allow for the rigorous examination of trait evolution in a tremendous diversity of organisms. Enemy driven adaptive radiation was a key prediction of Ehrlich and Raven's coevolutionary hypothesis [Ehrlich PR, Raven PH (1964) Evolution 18:586-608], yet has remained largely untested. Here we examine patterns of trait evolution in 51 North American milkweed species (Asclepias), using maximum likelihood methods. We study 7 traits of the milkweeds, ranging from seed size and foliar physiological traits to defense traits (cardenolides, latex, and trichomes) previously shown to impact herbivores, including the monarch butterfly. We compare the fit of simple random-walk models of trait evolution to models that incorporate stabilizing selection (Ornstein-Ulenbeck process), as well as time-varying rates of trait evolution. Early bursts of trait evolution were implicated for 2 traits, while stabilizing selection was implicated for several others. We further modeled the relationship between trait change and species diversification while allowing rates of trait evolution to vary during the radiation. Species-rich lineages underwent a proportionately greater decline in latex and cardenolides relative to species-poor lineages, and the rate of trait change was most rapid early in the radiation. An interpretation of this result is that reduced investment in defensive traits accelerated diversification, and disproportionately so, early in the adaptive radiation of milkweeds. PMID- 19805161 TI - Why data-sharing policies matter. PMID- 19805162 TI - Ancient origin of placental expression in the growth hormone genes of anthropoid primates. AB - In anthropoid primates, growth hormone (GH) genes have undergone at least 2 independent locus expansions, one in platyrrhines (New World monkeys) and another in catarrhines (Old World monkeys and apes). In catarrhines, the GH cluster has a pituitary-expressed gene called GH1; the remaining GH genes include placental GHs and placental lactogens. Here, we provide cDNA sequence evidence that the platyrrhine GH cluster also includes at least 3 placenta expressed genes and phylogenetic evidence that placenta expressed anthropoid GH genes have undergone strong adaptive evolution, whereas pituitary-expressed GH genes have faced strict functional constraint. Our phylogenetic evidence also points to lineage-specific gene gain and loss in early placental mammalian evolution, with at least three copies of the GH gene present at the time of the last common ancestor (LCA) of primates, rodents, and laurasiatherians. Anthropoid primates and laurasiatherians share gene descendants of one of these three copies, whereas rodents and strepsirrhine primates each maintain a separate copy. Eight of the amino-acid replacements that occurred on the lineage leading to the LCA of extant anthropoids have been implicated in GH signaling at the maternal-fetal interface. Thus, placental expression of GH may have preceded the separate series of GH gene duplications that occurred in catarrhines and platyrrhines (i.e., the roles played by placenta-expressed GHs in human pregnancy may have a longer evolutionary history than previously appreciated). PMID- 19805163 TI - Hutchinson's duality: the once and future niche. AB - The duality between "niche" and "biotope" proposed by G. Evelyn Hutchinson provides a powerful way to conceptualize and analyze biogeographical distributions in relation to spatial environmental patterns. Both Joseph Grinnell and Charles Elton had attributed niches to environments. Attributing niches, instead, to species, allowed Hutchinson's key innovation: the formal severing of physical place from environment that is expressed by the duality. In biogeography, the physical world (a spatial extension of what Hutchinson called the biotope) is conceived as a map, each point (or cell) of which is characterized by its geographical coordinates and the local values of n environmental attributes at a given time. Exactly the same n environmental attributes define the corresponding niche space, as niche axes, allowing reciprocal projections between the geographic distribution of a species, actual or potential, past or future, and its niche. In biogeographical terms, the realized niche has come to express not only the effects of species interactions (as Hutchinson intended), but also constraints of dispersal limitation and the lack of contemporary environments corresponding to parts of the fundamental niche. Hutchinson's duality has been used to classify and map environments; model potential species distributions under past, present, and future climates; study the distributions of invasive species; discover new species; and simulate increasingly more realistic worlds, leading to spatially explicit, stochastic models that encompass speciation, extinction, range expansion, and evolutionary adaptation to changing environments. PMID- 19805164 TI - Minimal dynamics and the classification of C*-algebras. AB - Let X be an infinite, compact, metrizable space of finite covering dimension and alpha: X --> X a minimal homeomorphism. We prove that the crossed product C(X) times sign, right closed(alpha) Z absorbs the Jiang-Su algebra tensorially and has finite nuclear dimension. As a consequence, these algebras are determined up to isomorphism by their graded ordered K-theory under the necessary condition that their projections separate traces. This result applies, in particular, to those crossed products arising from uniquely ergodic homeomorphisms. PMID- 19805165 TI - Endothelial nitric oxide synthase negatively regulates hydrogen peroxide stimulated AMP-activated protein kinase in endothelial cells. AB - Hydrogen peroxide and other reactive oxygen species are intimately involved in endothelial cell signaling. In many cell types, the AMP-activated protein kinase (AMPK) has been implicated in the control of metabolic responses, but the role of endothelial cell redox signaling in the modulation of AMPK remains to be completely defined. We used RNA interference and pharmacological methods to establish that H(2)O(2) is a critical activator of AMPK in cultured bovine aortic endothelial cells (BAECs). H(2)O(2) treatment of BAECs rapidly and significantly increases the phosphorylation of AMPK. The EC(50) for H(2)O(2)-promoted phosphorylation of AMPK is 65 + or - 15 microM, within the physiological range of cellular H(2)O(2) concentrations. The Ca(2+)/calmodulin-dependent protein kinase kinase-beta (CaMKKbeta) inhibitor STO-609 abolishes H(2)O(2)-dependent AMPK activation, whereas eNOS inhibitors enhance AMPK activation. Similarly, siRNA mediated knockdown of CaMKKbeta abrogates AMPK activation, whereas siRNA-mediated knockdown of eNOS leads to a striking increase in AMPK phosphorylation. Cellular imaging studies using the H(2)O(2) biosensor HyPer show that siRNA-mediated eNOS knockdown leads to a marked increase in intracellular H(2)O(2) generation, which is blocked by PEG-catalase. eNOS(-/-) mice show a marked increase in AMPK phosphorylation in liver and lung compared to wild-type mice. Lung endothelial cells from eNOS(-/-) mice also show a significant increase in AMPK phosphorylation. Taken together, these results establish that CaMKKbeta is critically involved in mediating the phosphorylation of AMPK promoted by H(2)O(2) in endothelial cells, and document that eNOS is an important negative regulator of AMPK phosphorylation and intracellular H(2)O(2) generation in endothelial cells. PMID- 19805166 TI - Cargo sorting into multivesicular bodies in vitro. AB - Genetic studies have identified a number of proteins required for the internalization of biosynthetic and endocytic cargo proteins transported to the multivesicular body (MVB). We have developed a cell-free reaction that recapitulates the internalization of a yeast biosynthetic membrane cargo protein, carboxypeptidase S (CPS), into the interior of an endosome. A recombinant form of CPS containing a biotinylation site from an Escherichia coli protein is accumulated in a vps27 yeast mutant blocked in the MVB internalization event. Endosomes isolated from the vps27 mutant are exposed to E. coli biotin ligase, which acts on only those CPS molecules with a cytosol-exposed N-terminal domain. Internalization of biotin-tagged CPS is measured by the detection of trypsin inaccessible, membrane-protected species. Biotinylated CPS internalization requires ATP and functional forms of Vps27p and Vps4p and depends on the availability of an exposed lysine residue critical for CPS ubiquitylation. PMID- 19805167 TI - Malignant cell-derived PlGF promotes normalization and remodeling of the tumor vasculature. AB - Vascular functions of PlGF remain poorly understood and controversial. Here, we show that tumor cell-derived PlGF-1 and PlGF-2 displayed significant remodeling effects on the tumor vasculature, leading to a normalized vascular phenotype and improved functions against leakage. In two murine tumor models, that is, T241 fibrosarcoma and Lewis lung carcinoma, stable expression of PlGF-1 and PlGF-2 in tumor cells resulted in significant reduction of tumor microvascular density and branch formation. Markedly, the vasculature in PlGF-expressing tumors consisted of relatively large-diameter microvessels with substantial improvement of pericyte coverage. Similarly, PlGF-induced vascular normalization and remodeling were also observed in a spontaneous human choriocarcinoma that expressed endogenous PlGF. Our findings shed light on functions of PlGF as a vascular remodeling factor that normalizes the tumor vasculature and thus may have conceptual implications of cancer therapy. PMID- 19805168 TI - Targeting proteins to distinct subcellular compartments reveals unique requirements for MHC class I and II presentation. AB - Peptides derived from exogenous proteins are presented by both MHC class I and II. Despite extensive study, the features of the endocytic pathway that mediate cross-presentation of exogenous antigens on MHC class I are not entirely understood and difficult to generalize to all proteins. Here, we used dendritic cells and macrophages to examine MHC class I and II presentation of hen egg-white lysozyme (HEL) in different forms, soluble and liposome encapsulated. Soluble HEL or HEL targeted to a late endosomal compartment only allowed for MHC class II presentation, in a process that was blocked by chloroquine and a cathepsin S (CatS) inhibitor; brefeldin A (BFA) also blocked presentation, indicating a requirement for nascent MHC class II. In contrast, liposome-encapsulated HEL targeted to early endosomes entered the MHC class I and II presentation pathways. Cross-presentation of HEL in early endosomal liposomes had several unique features: it was markedly increased by BFA and by blockade of the proteasome or CatS activity, it occurred independently of the transporter associated with antigen processing but required an MHC class I surface-stabilizing peptide, and it was inhibited by chloroquine. Remarkably, chloroquine facilitated MHC class I cross-presentation of soluble HEL and HEL in late endosomal liposomes. Altogether, MHC class I and II presentation of HEL occurred through pathways having distinct molecular and proteolytic requirements. Moreover, MHC class I sampled antigenic peptides from various points along the endocytic route. PMID- 19805169 TI - Enzyme millisecond conformational dynamics do not catalyze the chemical step. AB - The idea that enzymes catalyze reactions by dynamical coupling between the conformational motions and the chemical coordinates has recently attracted major experimental and theoretical interest. However, experimental studies have not directly established that the conformational motions transfer energy to the chemical coordinate, and simulating enzyme catalysis on the relevant timescales has been impractical. Here, we introduce a renormalization approach that transforms the energetics and dynamics of the enzyme to an equivalent low dimensional system, and allows us to simulate the dynamical coupling on a ms timescale. The simulations establish, by means of several independent approaches, that the conformational dynamics is not remembered during the chemical step and does not contribute significantly to catalysis. Nevertheless, the precise nature of this coupling is a question of great importance. PMID- 19805170 TI - The role of fluctuations and stress on the effective viscosity of cell aggregates. AB - Cell aggregates are a tool for in vitro studies of morphogenesis, cancer invasion, and tissue engineering. They respond to mechanical forces as a complex rather than simple liquid. To change an aggregate's shape, cells have to overcome energy barriers. If cell shape fluctuations are active enough, the aggregate spontaneously relaxes stresses ("fluctuation-induced flow"). If not, changing the aggregate's shape requires a sufficiently large applied stress ("stress-induced flow"). To capture this distinction, we develop a mechanical model of aggregates based on their cellular structure. At stress lower than a characteristic stress tau*, the aggregate as a whole flows with an apparent viscosity eta*, and at higher stress it is a shear-thinning fluid. An increasing cell-cell tension results in a higher eta* (and thus a slower stress relaxation time t(c)). Our constitutive equation fits experiments of aggregate shape relaxation after compression or decompression in which irreversibility can be measured; we find t(c) of the order of 5 h for F9 cell lines. Predictions also match numerical simulations of cell geometry and fluctuations. We discuss the deviations from liquid behavior, the possible overestimation of surface tension in parallel-plate compression measurements, and the role of measurement duration. PMID- 19805171 TI - Evidence for biological nitrification inhibition in Brachiaria pastures. AB - Nitrification, a key process in the global nitrogen cycle that generates nitrate through microbial activity, may enhance losses of fertilizer nitrogen by leaching and denitrification. Certain plants can suppress soil-nitrification by releasing inhibitors from roots, a phenomenon termed biological nitrification inhibition (BNI). Here, we report the discovery of an effective nitrification inhibitor in the root-exudates of the tropical forage grass Brachiaria humidicola (Rendle) Schweick. Named "brachialactone," this inhibitor is a recently discovered cyclic diterpene with a unique 5-8-5-membered ring system and a gamma-lactone ring. It contributed 60-90% of the inhibitory activity released from the roots of this tropical grass. Unlike nitrapyrin (a synthetic nitrification inhibitor), which affects only the ammonia monooxygenase (AMO) pathway, brachialactone appears to block both AMO and hydroxylamine oxidoreductase enzymatic pathways in Nitrosomonas. Release of this inhibitor is a regulated plant function, triggered and sustained by the availability of ammonium (NH(4)(+)) in the root environment. Brachialactone release is restricted to those roots that are directly exposed to NH(4)(+). Within 3 years of establishment, Brachiaria pastures have suppressed soil nitrifier populations (determined as amoA genes; ammonia-oxidizing bacteria and ammonia-oxidizing archaea), along with nitrification and nitrous oxide emissions. These findings provide direct evidence for the existence and active regulation of a nitrification inhibitor (or inhibitors) release from tropical pasture root systems. Exploiting the BNI function could become a powerful strategy toward the development of low-nitrifying agronomic systems, benefiting both agriculture and the environment. PMID- 19805172 TI - Institutional challenges for mining and sustainability in Peru. AB - Global consumption continues to generate growth in mining. In lesser developed economies, this growth offers the potential to generate new resources for development, but also creates challenges to sustainability in the regions in which extraction occurs. This context leads to debate on the institutional arrangements most likely to build synergies between mining, livelihoods, and development, and on the socio-political conditions under which such institutions can emerge. Building from a multiyear, three-country program of research projects, Peru, a global center of mining expansion, serves as an exemplar for analyzing the effects of extractive industry on livelihoods and the conditions under which arrangements favoring local sustainability might emerge. This program is guided by three emergent hypotheses in human-environmental sciences regarding the relationships among institutions, knowledge, learning, and sustainability. The research combines in-depth and comparative case study analysis, and uses mapping and spatial analysis, surveys, in-depth interviews, participant observation, and our own direct participation in public debates on the regulation of mining for development. The findings demonstrate the pressures that mining expansion has placed on water resources, livelihood assets, and social relationships. These pressures are a result of institutional conditions that separate the governance of mineral expansion, water resources, and local development, and of relationships of power that prioritize large scale investment over livelihood and environment. A further problem is the poor communication between mining sector knowledge systems and those of local populations. These results are consistent with themes recently elaborated in sustainability science. PMID- 19805173 TI - IL-18 binding protein-expressing mesenchymal stem cells improve myocardial protection after ischemia or infarction. AB - IL-18 is a proinflammatory cytokine known to cause tissue injury by inducing inflammation and cell death. Increased levels of IL-18 are associated with myocardial injury after ischemia or infarction. IL-18-binding protein (IL-18BP), the naturally occurring inhibitor of IL-18 activity, decreases the severity of inflammation in response to injury. In the present study, mesenchymal stem cells (MSCs) derived from mice transgenic for over expression of human IL-18BP were tested in rat models of global myocardial ischemia and acute myocardial infarction. Improved myocardial function is associated with production of VEGF, and in vitro, IL-18BP MSCs secreted higher levels of constitutive VEGF compared to wild-type MSCs. Whereas IL-18 increased cell death and reduced VEGF in wild type MSCs, IL-18BP MSCs were protected. In an isolated heart model, intracoronary infusion of IL-18BP MSCs before ischemia increased postischemic left ventricular (LV) developed pressure to 79.5 + or - 9.47 mmHg compared to 59.3 + or - 7.8 mmHg in wild-type MSCs and 37.8 + or - 5 mmHg in the vehicle group. Similarly, using a coronary artery ligation model, intramyocardial injection of IL-18BP MSCs improved LV ejection fraction to 67.8 + or - 1.76% versus wild-type MSCs (57.4 + or - 1.33%) and vehicle (39.2 + or - 2.07%), increased LV fractional shortening 1.25-fold over wild-type MSCs and 1.95-fold over vehicle, decreased infarct size to 38.8 + or - 2.16% compared to 46.4 + or - 1.92% in wild-type MSCs and 60.7 + or - 2.2% in vehicle, reduced adverse ventricular remodeling, increased myocardial VEGF production, and decreased IL-6 levels. This study provides the concept that IL-18BP genetically modified stem cells improve cardioprotection over that observed with unmodified stem cells. PMID- 19805174 TI - Reticulon 4B (Nogo-B) is necessary for macrophage infiltration and tissue repair. AB - Blood vessel formation during ischemia and wound healing requires coordination of the inflammatory response with genes that regulate blood vessel assembly. Here we show that the reticulon family member 4B, aka Nogo-B, is upregulated in response to ischemia and is necessary for blood flow recovery secondary to ischemia and wound healing. Mice lacking Nogo-B exhibit reduced arteriogenesis and angiogenesis that are linked to a decrease in macrophage infiltration and inflammatory gene expression in vivo. Bone marrow-derived macrophages isolated from Nogo knock-out mice have reduced spreading and chemotaxis due to impaired Rac activation. Bone marrow reconstitution experiments show that Nogo in myeloid cells is necessary to promote macrophage homing and functional recovery after limb ischemia. Thus, endogenous Nogo coordinates macrophage-mediated inflammation with arteriogenesis, wound healing, and blood flow control. PMID- 19805175 TI - Generalized antifungal activity and 454-screening of Pseudonocardia and Amycolatopsis bacteria in nests of fungus-growing ants. AB - In many host-microbe mutualisms, hosts use beneficial metabolites supplied by microbial symbionts. Fungus-growing (attine) ants are thought to form such a mutualism with Pseudonocardia bacteria to derive antibiotics that specifically suppress the coevolving pathogen Escovopsis, which infects the ants' fungal gardens and reduces growth. Here we test 4 key assumptions of this Pseudonocardia Escovopsis coevolution model. Culture-dependent and culture-independent (tag encoded 454-pyrosequencing) surveys reveal that several Pseudonocardia species and occasionally Amycolatopsis (a close relative of Pseudonocardia) co-occur on workers from a single nest, contradicting the assumption of a single pseudonocardiaceous strain per nest. Pseudonocardia can occur on males, suggesting that Pseudonocardia could also be horizontally transmitted during mating. Pseudonocardia and Amycolatopsis secretions kill or strongly suppress ant cultivated fungi, contradicting the previous finding of a growth-enhancing effect of Pseudonocardia on the cultivars. Attine ants therefore may harm their own cultivar if they apply pseudonocardiaceous secretions to actively growing gardens. Pseudonocardia and Amycolatopsis isolates also show nonspecific antifungal activities against saprotrophic, endophytic, entomopathogenic, and garden-pathogenic fungi, contrary to the original report of specific antibiosis against Escovopsis alone. We conclude that attine-associated pseudonocardiaceous bacteria do not exhibit derived antibiotic properties to specifically suppress Escovopsis. We evaluate hypotheses on nonadaptive and adaptive functions of attine integumental bacteria, and develop an alternate conceptual framework to replace the prevailing Pseudonocardia-Escovopsis coevolution model. If association with Pseudonocardia is adaptive to attine ants, alternate roles of such microbes could include the protection of ants or sanitation of the nest. PMID- 19805176 TI - Distinct immune responses to transgene products from rAAV1 and rAAV8 vectors. AB - Recently developed serotypes of recombinant adeno-associated virus (rAAV) vectors have significantly enhanced the use of rAAV vectors for gene therapy. However, host immune responses to the transgene products from different serotypes remain uncharacterized. In the present study, we evaluated the differential immune responses to the transgene products from rAAV1 and rAAV8 vectors. In non-obese diabetic (NOD) mice, which have a hypersensitive immunity, rAAV serotype 1 vector (rAAV1-hAAT) induced high levels of both humoral and cellular responses, while rAAV8-hAAT did not. In vitro studies showed that rAAV1, but not rAAV8 vector transduced dendritic cells (DCs) efficiently. In vivo studies indicated that vector transduction of DCs was essential for the immune responses; while the presence of a transgene product (or foreign gene product produced by host cells) was not immunogenic. Intriguingly, preimmunization with rAAV8-hAAT vector or with serum of hAAT transgenic NOD mouse induced immune tolerance to rAAV1-hAAT injection. These results demonstrate the immunogenic differences of rAAV1 and rAAV8 and imply tremendous potential for these vectors in different applications, where an immune response to transgene is to be either elicited or avoided. PMID- 19805177 TI - Hyperpolarized xenon NMR and MRI signal amplification by gas extraction. AB - A method is reported for enhancing the sensitivity of NMR of dissolved xenon by detecting the signal after extraction to the gas phase. We demonstrate hyperpolarized xenon signal amplification by gas extraction (Hyper-SAGE) in both NMR spectra and magnetic resonance images with time-of-flight information. Hyper SAGE takes advantage of a change in physical phase to increase the density of polarized gas in the detection coil. At equilibrium, the concentration of gas phase xenon is approximately 10 times higher than that of the dissolved-phase gas. After extraction the xenon density can be further increased by several orders of magnitude by compression and/or liquefaction. Additionally, being a remote detection technique, the Hyper-SAGE effect is further enhanced in situations where the sample of interest would occupy only a small proportion of the traditional NMR receiver. Coupled with targeted xenon biosensors, Hyper-SAGE offers another path to highly sensitive molecular imaging of specific cell markers by detection of exhaled xenon gas. PMID- 19805178 TI - The niche, limits to species' distributions, and spatiotemporal variation in demography across the elevation ranges of two monkeyflowers. AB - Understanding the processes that create and maintain species' geographic range limits has implications for many questions in ecology, evolution, and conservation biology. Many expectations for the ecological and evolutionary dynamics of populations at the range margin rest on the concordance of geographic limits and the limits of a species' ecological niche. If range limits are coincident with niche limits, then marginal populations should have lower and/or more variable vital rates and population growth rates than central populations. Using data from 8 annual censuses of marked individuals, I investigated the demography of Mimulus cardinalis and Mimulus lewisii across the species' elevation ranges. Central and marginal populations exhibited striking demographic differences, but only for one species were differences in expected directions. Marginal populations from the M. lewisii lower elevation range limit had lower and more variable survival than central populations and appeared to be demographic sinks. In contrast, marginal populations from the M. cardinalis upper elevation limit had higher fecundity and higher population growth rates than central populations. Although the species differed with respect to central marginal patterns, they were concordant with respect to elevation; that is, both species had higher fitness in higher reaches of their examined ranges. Potential explanations for these patterns include source-sink dynamics, with asymmetrical gene flow mediated by river currents, and climate change, with recent warming shifting the species' climatic envelopes to higher elevations. Hence, assessment of spatiotemporal variation in both demography and dispersal is necessary to fully understand the relationship between the niche and species' distributions. PMID- 19805179 TI - Niches, body sizes, and the disassembly of mammal communities on the Sunda Shelf islands. AB - The rising sea level at the end of the Pleistocene that created the islands of the Sunda Shelf in Indonesia and Malaysia provides a natural experiment in community disassembly and offers insights into the effects of body size and niches on abundance, distribution, and diversity. Since isolation, terrestrial mammal communities of these islands have been reduced by extinction, with virtually no offsetting colonization. We document three empirical patterns of disassembly, all of which are significantly different from null models of random assembly: (i) a diversity-area relationship: the number of taxa is strongly and positively correlated with island area; (ii) nested subset composition: species that occur on small islands tend to be subsets of more diverse communities inhabiting larger islands; and (iii) body size distributions: species of intermediate body sizes occur on the greatest number of islands, and smaller islands have smaller ranges of body sizes, caused by the absence of species of both very large and extremely small size. These patterns reveal the role of body size and other niche characteristics, such as habitat requirements and trophic status, in the differential susceptibility of taxa to extinction. PMID- 19805180 TI - Redox trapping of arsenic during groundwater discharge in sediments from the Meghna riverbank in Bangladesh. AB - Groundwater arsenic (As) is elevated in the shallow Holocene aquifers of Bangladesh. In the dry season, the shallow groundwater discharges to major rivers. This process may influence the chemistry of the river and the hyporheic zone sediment. To assess the fate of As during discharge, surface (0-5 cm) and subsurface (1-3 m) sediment samples were collected at 9 sites from the bank of the Meghna River along a transect from its northern source (25 degrees N) to the Bay of Bengal (22.5 degrees N). Bulk As concentrations of surface sediment averaged 16 +/- 7 mg/kg (n = 9). Subsurface sediment contained higher mean concentrations of As of 4,000 mg/kg (n = 14), ranging from 1 to 23,000 mg/kg As, with >100 mg/kg As measured at 8 sites. X-ray absorption near-edge structure spectroscopy indicated that As was mainly arsenate and arsenite, not As-bearing sulfides. We hypothesize that the elevated sediment As concentrations form as As rich groundwater discharges to the river, and enters a more oxidizing environment. A significant portion of dissolved As sorbs to iron-bearing minerals, which form a natural reactive barrier. Recycling of this sediment-bound As to the Ganges-Brahmaputra-Meghna Delta aquifer provides a potential source of As to further contaminate groundwater. Furthermore, chemical fluxes from groundwater discharge from the Ganges-Brahmaputra-Meghna Delta may be less than previous estimates because this barrier can immobilize many elements. PMID- 19805181 TI - The Corynebacterium diphtheriae shaft pilin SpaA is built of tandem Ig-like modules with stabilizing isopeptide and disulfide bonds. AB - Cell-surface pili are important virulence factors that enable bacterial pathogens to adhere to specific host tissues and modulate host immune response. Relatively little is known about the structure of Gram-positive bacterial pili, which are built by the sortase-catalyzed covalent crosslinking of individual pilin proteins. Here we report the 1.6-A resolution crystal structure of the shaft pilin component SpaA from Corynebacterium diphtheriae, revealing both common and unique features. The SpaA pilin comprises 3 tandem Ig-like domains, with characteristic folds related to those typically found in non-pilus adhesins. Whereas both the middle and the C-terminal domains contain an intramolecular Lys Asn isopeptide bond, previously detected in the shaft pilins of Streptococcus pyogenes and Bacillus cereus, the middle Ig-like domain also harbors a calcium ion, and the C-terminal domain contains a disulfide bond. By mass spectrometry, we show that the SpaA monomers are cross-linked in the assembled pili by a Lys Thr isopeptide bond, as predicted by previous genetic studies. Together, our results reveal that despite profound dissimilarities in primary sequences, the shaft pilins of Gram-positive pathogens have strikingly similar tertiary structures, suggesting a modular backbone construction, including stabilizing intermolecular and intramolecular isopeptide bonds. PMID- 19805182 TI - The Tor and PKA signaling pathways independently target the Atg1/Atg13 protein kinase complex to control autophagy. AB - Macroautophagy (or autophagy) is a conserved degradative pathway that has been implicated in a number of biological processes, including organismal aging, innate immunity, and the progression of human cancers. This pathway was initially identified as a cellular response to nutrient deprivation and is essential for cell survival during these periods of starvation. Autophagy is highly regulated and is under the control of a number of signaling pathways, including the Tor pathway, that coordinate cell growth with nutrient availability. These pathways appear to target a complex of proteins that contains the Atg1 protein kinase. The data here show that autophagy in Saccharomyces cerevisiae is also controlled by the cAMP-dependent protein kinase (PKA) pathway. Elevated levels of PKA activity inhibited autophagy and inactivation of the PKA pathway was sufficient to induce a robust autophagy response. We show that in addition to Atg1, PKA directly phosphorylates Atg13, a conserved regulator of Atg1 kinase activity. This phosphorylation regulates Atg13 localization to the preautophagosomal structure, the nucleation site from which autophagy pathway transport intermediates are formed. Atg13 is also phosphorylated in a Tor-dependent manner, but these modifications appear to occur at positions distinct from the PKA phosphorylation sites identified here. In all, our data indicate that the PKA and Tor pathways function independently to control autophagy in S. cerevisiae, and that the Atg1/Atg13 kinase complex is a key site of signal integration within this degradative pathway. PMID- 19805183 TI - The evolution of antiherbivore defenses and their contribution to species coexistence in the tropical tree genus Inga. AB - Plants and their herbivores constitute more than half of the organisms in tropical forests. Therefore, a better understanding of the evolution of plant defenses against their herbivores may be central for our understanding of tropical biodiversity. Here, we address the evolution of antiherbivore defenses and their possible contribution to coexistence in the Neotropical tree genus Inga (Fabaceae). Inga has >300 species, has radiated recently, and is frequently one of the most diverse and abundant genera at a given site. For 37 species from Panama and Peru we characterized developmental, ant, and chemical defenses against herbivores. We found extensive variation in defenses, but little evidence of phylogenetic signal. Furthermore, in a multivariate analysis, developmental, ant, and chemical defenses varied independently (were orthogonal) and appear to have evolved independently of each other. Our results are consistent with strong selection for divergent defensive traits, presumably mediated by herbivores. In an analysis of community assembly, we found that Inga species co-occurring as neighbors are more different in antiherbivore defenses than random, suggesting that possessing a rare defense phenotype increases fitness. These results imply that interactions with herbivores may be an important axis of niche differentiation that permits the coexistence of many species of Inga within a single site. Interactions between plants and their herbivores likely play a key role in the generation and maintenance of the conspicuously high plant diversity in the tropics. PMID- 19805184 TI - Traffic-driven epidemic spreading in finite-size scale-free networks. AB - The study of complex networks sheds light on the relation between the structure and function of complex systems. One remarkable result is the absence of an epidemic threshold in infinite-size, scale-free networks, which implies that any infection will perpetually propagate regardless of the spreading rate. The vast majority of current theoretical approaches assumes that infections are transmitted as a reaction process from nodes to all neighbors. Here we adopt a different perspective and show that the epidemic incidence is shaped by traffic flow conditions. Specifically, we consider the scenario in which epidemic pathways are defined and driven by flows. Through extensive numerical simulations and theoretical predictions, it is shown that the value of the epidemic threshold in scale-free networks depends directly on flow conditions, in particular on the first and second moments of the betweenness distribution given a routing protocol. We consider the scenarios in which the delivery capability of the nodes is bounded or unbounded. In both cases, the threshold values depend on the traffic and decrease as flow increases. Bounded delivery provokes the emergence of congestion, slowing down the spreading of the disease and setting a limit for the epidemic incidence. Our results provide a general conceptual framework for the understanding of spreading processes on complex networks. PMID- 19805185 TI - Rational modulation of conformational fluctuations in adenylate kinase reveals a local unfolding mechanism for allostery and functional adaptation in proteins. AB - Elucidating the complex interplay between protein structure and dynamics is a prerequisite to an understanding of both function and adaptation in proteins. Unfortunately, it has been difficult to experimentally decouple these effects because it is challenging to rationally design mutations that will either affect the structure but not the dynamics, or that will affect the dynamics but not the structure. Here we adopt a mutation approach that is based on a thermal adaptation strategy observed in nature, and we use it to study the binding interaction of Escherichia coli adenylate kinase (AK). We rationally design several single-site, surface-exposed glycine mutations to selectively perturb the excited state conformational repertoire, leaving the ground-state X-ray crystallographic structure unaffected. The results not only demonstrate that the conformational ensemble of AK is significantly populated by a locally unfolded state that is depopulated upon binding, but also that the excited-state conformational ensemble can be manipulated through mutation, independent of perturbations of the ground-state structures. The implications of these results are twofold. First, they indicate that it is possible to rationally design dynamic allosteric mutations, which do not propagate through a pathway of structural distortions connecting the mutated and the functional sites. Secondly and equally as important, the results reveal a general strategy for thermal adaptation that allows enzymes to modulate binding affinity by controlling the amount of local unfolding in the native-state ensemble. These findings open new avenues for rational protein design and fundamentally illuminate the role of local unfolding in function and adaptation. PMID- 19805186 TI - Long-term risk preference and suboptimal decision making following adolescent alcohol use. AB - Individuals who abused alcohol at an early age show decision-making impairments. However, the question of whether maladaptive choice constitutes a predisposing factor to, or a consequence resulting from, alcohol exposure remains open. To examine whether a causal link exists between voluntary alcohol consumption during adolescence and adult decision making the present studies used a rodent model. High levels of voluntary alcohol intake were promoted by providing adolescent rats with access to alcohol in a palatable gel matrix under nondeprivation conditions. A probability-discounting instrumental response task offered a choice between large but uncertain rewards and small but certain rewards to assess risk based choice in adulthood either 3 weeks or 3 months following alcohol exposure. While control animals' performance on this task closely conformed to a predictive model of risk-neutral value matching, rats that consumed high levels of alcohol during adolescence violated this model, demonstrating greater risk preference. Evidence of significant risk bias was still present when choice was assessed 3 months following discontinuation of alcohol access. These findings provide evidence that adolescent alcohol exposure may lead to altered decision making during adulthood and this model offers a promising approach to the investigation of the neurobiological underpinnings of this link. PMID- 19805187 TI - Upregulation of three Drosophila homologs of human chromosome 21 genes alters synaptic function: implications for Down syndrome. AB - At the neuronal level of Down syndrome (DS) brains, there are evidences of altered shape, number, and density of synapses, as well as aberrant endocytosis associated with accumulation of enlarged endosomes, suggesting that proteins involved in synaptic vesicle recycling may play key roles in DS neurons. However, the exact mechanism underlying those anomalies is not well understood. We hypothesize that overexpression of three genes, dap160/itsn1, synj/synj1, and nla/dscr1, located on human chromosome 21 play important roles in DS neurons. Here, we systematically investigate the effects of multiple gene overexpression on synaptic morphology and endocytosis to identify possible dominant gene or genes. We found that overexpression of individual genes lead to abnormal synaptic morphology, but all three genes are necessary to cause impaired vesicle recycling and affect locomotor vigor. Furthermore, we report that dap160 overexpression alters the subcellular distribution of synaptojanin, and overexpression of nla regulates the phosphoinositol 5' phosphatase activity of synaptojanin. These findings imply that restoring the level of any one of these genes may reduce endocytic defects seen in DS. PMID- 19805188 TI - Melanin-concentrating hormone directly inhibits GnRH neurons and blocks kisspeptin activation, linking energy balance to reproduction. AB - A link between energy balance and reproduction is critical for the survival of all species. Energy-consuming reproductive processes need to be aborted in the face of a negative energy balance, yet knowledge of the pathways mediating this link remains limited. Fasting and food restriction that inhibit fertility also upregulate the hypothalamic melanin-concentrating hormone (MCH) system that promotes feeding and decreases energy expenditure; MCH knockout mice are lean and have a higher metabolism but remain fertile. MCH also modulates sleep, drug abuse behavior, and mood, and MCH receptor antagonists are currently being developed as antiobesity and antidepressant drugs. Despite the clinical implications of MCH, the direct postsynaptic effects of MCH have never been reported in CNS neurons. Using patch-clamp recordings in brain slices from multiple lines of transgenic GFP mice, we demonstrate a strong inhibitory effect of MCH on an exclusive population of septal vGluT2-GnRH neurons that is activated by the puberty triggering and preovulatory luteinizing hormone surge-mediating peptide, kisspeptin. MCH has no effect on kisspeptin-insensitive GnRH, vGluT2, cholinergic, or GABAergic neurons located within the same nucleus. The inhibitory effects of MCH are reproducible and nondesensitizing and are mediated via a direct postsynaptic Ba(2+)-sensitive K(+) channel mechanism involving the MCHR1 receptor. MCH immunoreactive fibers are in close proximity to vGluT2-GFP and GnRH GFP neurons. Importantly, MCH blocks the excitatory effect of kisspeptin on vGluT2-GnRH neurons. Considering the role of MCH in regulating energy balance and of GnRH and kisspeptin in triggering puberty and maintaining fertility, MCH may provide a critical link between energy balance and reproduction directly at the level of the kisspeptin-activated vGluT2-GnRH neuron. PMID- 19805189 TI - Mice with the CHEK2*1100delC SNP are predisposed to cancer with a strong gender bias. AB - The CHEK2 kinase (Chk2 in mouse) is a member of a DNA damage response pathway that regulates cell cycle arrest at cell cycle checkpoints and facilitates the repair of dsDNA breaks by a recombination-mediated mechanism. There are numerous variants of the CHEK2 gene, at least one of which, CHEK2*1100delC (SNP), associates with breast cancer. A mouse model in which the wild-type Chk2 has been replaced by a Chk2*1100delC allele was tested for elevated risk of spontaneous cancer and increased sensitivity to challenge by a carcinogenic compound. Mice homozygous for Chk2*1100delC produced more tumors than wild-type mice, whereas heterozygous mice were not statistically different. When fractionated by gender, however, homozygous and heterozygous mice developed spontaneous tumors more rapidly and to a far greater extent than wild-type mice, indicative of a marked gender bias in mice harboring the variant allele. Consistent with our previous data showing elevated genomic instability in mouse embryonic fibroblasts (MEFs) derived from mice homozygous for Chk2*1100delC, the level of Cdc25A was elevated in heterozygous and homozygous MEFs and tumors. When challenged with the carcinogen 7,12-dimethylbenz[a]anthracene, all mice, regardless of genotype, had a reduced lifespan. Latency for mammary tumorigenesis was reduced significantly in mice homozygous for Chk2*1100delC but unexpectedly increased for the development of lymphomas. An implication from this study is that individuals who harbor the variant CHEK2*1100delC allele not only are at an elevated risk for the development of cancer but also that this risk can be further increased as a result of environmental exposure. PMID- 19805190 TI - Loss of the plant DEAD-box protein ISE1 leads to defective mitochondria and increased cell-to-cell transport via plasmodesmata. AB - Plants have intercellular channels, plasmodesmata (PD), that span the cell wall to enable cell-to-cell transport of micro- and macromolecules. We identified an Arabidopsis thaliana embryo lethal mutant increased size exclusion limit 1 (ise1) that results in increased PD-mediated transport of fluorescent tracers. The ise1 mutants have a higher frequency of branched and twinned PD than wild-type embryos. Silencing of ISE1 in mature Nicotiana benthamiana leaves also leads to increased PD transport, as monitored by intercellular movement of a GFP fusion to the tobacco mosaic virus movement protein. ISE1 encodes a putative plant-specific DEAD-box RNA helicase that localizes specifically to mitochondria. The N-terminal 100 aa of ISE1 specify mitochondrial targeting. Mitochondrial metabolism is compromised severely in ise1 mutant embryos, because their mitochondrial proton gradient is disrupted and reactive oxygen species production is increased. Although mitochondria are essential for numerous cell-autonomous functions, the present studies demonstrate that mitochondrial function also regulates the critical cell non-cell-autonomous function of PD. PMID- 19805191 TI - Regulation of axonal development by natriuretic peptide hormones. AB - Natriuretic peptides (NPs) are a family of cardiac- and vascular-derived hormones that are well known for regulating blood pressure, but their expression in the brain poses an intriguing yet unanswered question concerning their roles in the nervous system. Here, we report several unique activities of these hormones in regulating axonal development of dorsal root ganglion (DRG) neurons in the spinal cord. First, the C-type NP (CNP) is expressed in a restricted area of the dorsal spinal cord and provides a cue that is necessary for bifurcation of central sensory afferents. Second, in the culture of embryonic DRG neurons, CNP stimulates branch formation, induces axon outgrowth, and attracts growth cones. Furthermore, these activities are mediated by cyclic guanosine-3',5' monophosphate (cGMP) signaling and can be elicited by other members of NP hormones. Thus, NPs represent a new class of extracellular factors that regulate key axonal processes during development. Because their receptors are present in many regions of the embryonic and adult brain, we propose that these hormones have wide influence on the development and function of the nervous system. PMID- 19805192 TI - Acriflavine inhibits HIF-1 dimerization, tumor growth, and vascularization. AB - HIF-1 is a heterodimeric transcription factor that mediates adaptive responses to hypoxia and plays critical roles in cancer progression. Using a cell-based screening assay we have identified acriflavine as a drug that binds directly to HIF-1alpha and HIF-2alpha and inhibits HIF-1 dimerization and transcriptional activity. Pretreatment of mice bearing prostate cancer xenografts with acriflavine prevented tumor growth and treatment of mice bearing established tumors resulted in growth arrest. Acriflavine treatment inhibited intratumoral expression of angiogenic cytokines, mobilization of angiogenic cells into peripheral blood, and tumor vascularization. These results provide proof of principle that small molecules can inhibit dimerization of HIF-1 and have potent inhibitory effects on tumor growth and vascularization. PMID- 19805193 TI - Structure of a trimeric nucleoporin complex reveals alternate oligomerization states. AB - The heptameric Nup84 complex constitutes an evolutionarily conserved building block of the nuclear pore complex. Here, we present the crystal structure of the heterotrimeric Sec13 x Nup145C x Nup84 complex, the centerpiece of the heptamer, at 3.2-A resolution. Nup84 forms a U-shaped alpha-helical solenoid domain, topologically similar to two other members of the heptamer, Nup145C and Nup85. The interaction between Nup84 and Nup145C is mediated via a hydrophobic interface located in the kink regions of the two solenoids that is reinforced by additional interactions of two long Nup84 loops. The Nup84 binding site partially overlaps with the homo-dimerization interface of Nup145C, suggesting competing binding events. Fitting of the elongated Z-shaped heterotrimer into electron microscopy (EM) envelopes of the heptamer indicates that structural changes occur at the Nup145C x Nup84 interface. Docking the crystal structures of all heptamer components into the EM envelope constitutes a major advance toward the completion of the structural characterization of the Nup84 complex. PMID- 19805194 TI - An evolutionary conserved function of the JAK-STAT pathway in anti-dengue defense. AB - Here, we show that the major mosquito vector for dengue virus uses the JAK-STAT pathway to control virus infection. Dengue virus infection in Aedes aegypti mosquitoes activates the JAK-STAT immune signaling pathway. The mosquito's susceptibility to dengue virus infection increases when the JAK-STAT pathway is suppressed through RNAi depletion of its receptor Domeless (Dome) and the Janus kinase (Hop), whereas mosquitoes become more resistant to the virus when the negative regulator of the JAK-STAT pathway, PIAS, is silenced. The JAK-STAT pathway exerts its anti-dengue activity presumably through one or several STAT regulated effectors. We have identified, and partially characterized, two JAK STAT pathway-regulated and infection-responsive dengue virus restriction factors (DVRFs) that contain putative STAT-binding sites in their promoter regions. Our data suggest that the JAK-STAT pathway is part of the A. aegypti mosquito's anti dengue defense and may act independently of the Toll pathway and the RNAi mediated antiviral defenses. PMID- 19805195 TI - Color-tuned neurons are spatially clustered according to color preference within alert macaque posterior inferior temporal cortex. AB - Large islands of extrastriate cortex that are enriched for color-tuned neurons have recently been described in alert macaque using a combination of functional magnetic resonance imaging (fMRI) and single-unit recording. These millimeter sized islands, dubbed "globs," are scattered throughout the posterior inferior temporal cortex (PIT), a swath of brain anterior to area V3, including areas V4, PITd, and posterior TEO. We investigated the micro-organization of neurons within the globs. We used fMRI to identify the globs and then used MRI-guided microelectrodes to test the color properties of single glob cells. We used color stimuli that sample the CIELUV perceptual color space at regular intervals to test the color tuning of single units, and make two observations. First, color tuned neurons of various color preferences were found within single globs. Second, adjacent glob cells tended to have the same color tuning, demonstrating that glob cells are clustered by color preference and suggesting that they are arranged in color columns. Neurons separated by 50 microm, measured parallel to the cortical sheet, had more similar color tuning than neurons separated by 100 microm, suggesting that the scale of the color columns is <100 microm. These results show that color-tuned neurons in PIT are organized by color preference on a finer scale than the scale of single globs. Moreover, the color preferences of neurons recorded sequentially along a given electrode penetration shifted gradually in many penetrations, suggesting that the color columns are arranged according to a chromotopic map reflecting perceptual color space. PMID- 19805196 TI - Increased expression of Mg(2+) transport proteins enhances the survival of Salmonella enterica at high temperature. AB - Mg(2+) homeostasis is important for Salmonella pathogenesis. In Salmonella enterica, the transcription of the mgtA gene, which encodes a Mg(2+) transporter, is regulated by a Mg(2+)-sensing riboswitch [Cromie MJ, Shi Y, Latifi T, Groisman EA (2006) Cell 125:71-84]. In a genetic analysis of the determinants of thermotolerance in S. enterica serovar Typhimurium, we isolated the chr-1 mutation that increased the resistance of exponential phase cells to killing by high temperature. This mutation is a single base change in the mgtA riboswitch that causes high-level constitutive expression of mgtA. We showed that another mgtA riboswitch mutation, DeltaUTR(re-100), which had been constructed by Cromie et al., also confers similar increased thermotolerance. Surprisingly, the chr-1 mutation is located at a position that would not be predicted to be important for the regulatory function of the riboswitch. We obtained physiological evidence suggesting that the chr-1 mutation increases the cytosolic free Mg(2+) concentration. High-level expression of the heterologous MgtE Mg(2+) transport protein of Bacillus subtilis also enhanced the thermotolerance of S. enterica. We hypothesize that increased Mg(2+) accumulation might enhance thermotolerance by protecting the integrity of proteins or membranes, by mitigating oxidative damage or acting as an inducer of thermoprotective functions. PMID- 19805197 TI - Layer-specific network oscillation and spatiotemporal receptive field in the visual cortex. AB - A quintessential feature of the neocortex is its laminar organization, and characterizing the activity patterns in different layers is an important step in understanding cortical processing. Using in vivo whole-cell recordings in rat visual cortex, we show that the temporal patterns of ongoing synaptic inputs to pyramidal neurons exhibit clear laminar specificity. Although low-frequency (approximately 2 Hz) activity is widely observed in layer 2/3 (L2/3), a narrow band fast oscillation (10-15 Hz) is prominent in layer 5 (L5). This fast oscillation is carried exclusively by excitatory inputs. Moreover, the frequency of ongoing activity is strongly correlated with the spatiotemporal window of visual integration: Neurons with fast-oscillating spontaneous inputs exhibit transient visual responses and small receptive fields (RFs), whereas those with slow inputs show prolonged responses and large RFs. These findings suggest that the neural representation of visual information within each layer is strongly influenced by the temporal dynamics of the local network manifest in spontaneous activity. PMID- 19805198 TI - Structure of an integrin alphaIIb beta3 transmembrane-cytoplasmic heterocomplex provides insight into integrin activation. AB - Heterodimeric integrin adhesion receptors regulate diverse biological processes including angiogenesis, thrombosis and wound healing. The transmembrane cytoplasmic domains (TMCDs) of integrins play a critical role in controlling activation of these receptors via an inside-out signaling mechanism, but the precise structural basis remains elusive. Here, we present the solution structure of integrin alphaIIb beta3 TMCD heterodimer, which reveals a right-handed coiled coil conformation with 2 helices intertwined throughout the transmembrane region. The helices extend into the cytoplasm and form a clasp that differs significantly from a recently published alphaIIb beta3 TMCD structure. We show that while a point mutation in the clasp interface modestly activates alphaIIb beta3, additional mutations in the transmembrane interface have a synergistic effect, leading to extensive integrin activation. Detailed analyses and structural comparison with previous studies suggest that extensive integrin activation is a highly concerted conformational transition process, which involves transmembrane coiled-coil unwinding that is triggered by the membrane-mediated alteration and disengagement of the membrane-proximal clasp. Our results provide atomic insight into a type I transmembrane receptor heterocomplex and the mechanism of integrin inside-out transmembrane signaling. PMID- 19805199 TI - Monkey drumming reveals common networks for perceiving vocal and nonvocal communication sounds. AB - Salient sounds such as those created by drumming can serve as means of nonvocal acoustic communication in addition to vocal sounds. Despite the ubiquity of drumming across human cultures, its origins and the brain regions specialized in processing such signals remain unexplored. Here, we report that an important animal model for vocal communication, the macaque monkey, also displays drumming behavior, and we exploit this finding to show that vocal and nonvocal communication sounds are represented by overlapping networks in the brain's temporal lobe. Observing social macaque groups, we found that these animals use artificial objects to produce salient periodic sounds, similar to acoustic gestures. Behavioral tests confirmed that these drumming sounds attract the attention of listening monkeys similarly as conspecific vocalizations. Furthermore, in a preferential looking experiment, drumming sounds influenced the way monkeys viewed their conspecifics, suggesting that drumming serves as a multimodal signal of social dominance. Finally, by using high-resolution functional imaging we identified those brain regions preferentially activated by drumming sounds or by vocalizations and found that the representations of both these communication sounds overlap in caudal auditory cortex and the amygdala. The similar behavioral responses to drumming and vocal sounds, and their shared neural representation, suggest a common origin of primate vocal and nonvocal communication systems and support the notion of a gestural origin of speech and music. PMID- 19805200 TI - Thrombospondin-1 is a critical effector of oncosuppressive activity of sst2 somatostatin receptor on pancreatic cancer. AB - The somatostatin receptor subtype 2 (sst2) behaves as a tumor suppressor when expressed and stimulated by its ligand somatostatin in pancreatic cancer. We reveal a mechanism underlying oncosuppressive action of sst2, whereby this inhibitory receptor upregulates the expression of the secreted angioinhibitory factor thrombospondin-1 (TSP-1), as demonstrated in exocrine BxPC-3 and endocrine BON pancreatic cancer cells. The sst2-dependent upregulation of TSP-1 occurs through the inhibition of the PI3K pathway. It depends on transcriptional and translational events, involving a previously undescribed IRES in the 5'-UTR of TSP-1 mRNA. Chick chorioallantoic membrane was used as an in vivo model to demonstrate that TSP-1 is a critical effector of the inhibitory role of sst2 on the neoangiogenesis and oncogenesis induced by pancreatic cancer cells. TSP-1 reduced in vitro tubulogenesis of endothelial cells when grown in conditioned medium from pancreatic cancer cells expressing sst2, as compared to those expressing the control vector. TSP-1 inhibited tumor cell-induced neoangiogenesis by directly sequestering the proangiogenic factor VEGF, and inactivating the angiogenesis initiated by VEGFR2 phosphorylation in endothelial cells. Using human pancreatic tissue-microarrays, the expression of both sst2 and TSP-1 was shown to be correlated during the pancreatic neoplastic program. Both proteins are nearly undetectable in normal exocrine pancreas and in most invasive cancer lesions, but their expression is strikingly upregulated in most preinvasive cancer-adjacent lesions. The upregulation of both sst2 and TSP-1 tumor suppressors may function as an early negative feedback to restrain pancreatic carcinogenesis. PMID- 19805201 TI - A genetic perspective on gender, culture, and mathematics performance. PMID- 19805202 TI - Gender, biology, and mathematics performance. PMID- 19805203 TI - No evidence for disease-like processes in fetal transplants. PMID- 19805206 TI - The trophic tapestry of the sea. PMID- 19805207 TI - Breadth-biased versus focused cognitive control in media multitasking behaviors. PMID- 19805208 TI - Stem cells and a cure for type 1 diabetes? PMID- 19805209 TI - Damage control in host-pathogen interactions. PMID- 19805210 TI - The genomic basis of trophic strategy in marine bacteria. AB - Many marine bacteria have evolved to grow optimally at either high (copiotrophic) or low (oligotrophic) nutrient concentrations, enabling different species to colonize distinct trophic habitats in the oceans. Here, we compare the genome sequences of two bacteria, Photobacterium angustum S14 and Sphingopyxis alaskensis RB2256, that serve as useful model organisms for copiotrophic and oligotrophic modes of life and specifically relate the genomic features to trophic strategy for these organisms and define their molecular mechanisms of adaptation. We developed a model for predicting trophic lifestyle from genome sequence data and tested >400,000 proteins representing >500 million nucleotides of sequence data from 126 genome sequences with metagenome data of whole environmental samples. When applied to available oceanic metagenome data (e.g., the Global Ocean Survey data) the model demonstrated that oligotrophs, and not the more readily isolatable copiotrophs, dominate the ocean's free-living microbial populations. Using our model, it is now possible to define the types of bacteria that specific ocean niches are capable of sustaining. PMID- 19805211 TI - Universal non-Landau, self-organized, lattice disordering percolative dopant network sub-T(c) phase transition in ceramic superconductors. AB - Ceramic superconductors (cuprates, pnictides, etc.) exhibit universal features in both T(c)(max) and in their planar lattice disordering measured by EXAFS, as reflected by three phase transitions. The two highest temperature transitions are known to be associated with formation of Jahn-Teller pseudogaps and superconductive gaps, with corresponding Landau order parameters, but no new gap is associated with the third transition below T(c), and its origin is mysterious. It is argued that the third subT(c) transition is a dopant glass transition, which is remarkably similar to topological transitions previously observed in chalcogenide and oxide alloy network glasses (like window glass). PMID- 19805212 TI - Fluorescent chlorophyll catabolites in bananas light up blue halos of cell death. AB - Breakdown of chlorophyll is a major contributor to the diagnostic color changes in fall leaves, and in ripening apples and pears, where it commonly provides colorless, nonfluorescent tetrapyrroles. In contrast, in ripening bananas (Musa acuminata) chlorophylls fade to give unique fluorescent catabolites (FCCs), causing yellow bananas to glow blue, when observed under UV light. Here, we demonstrate the capacity of the blue fluorescent chlorophyll catabolites to signal symptoms of programmed cell death in a plant. We report on studies of bright blue luminescent rings on the peel of very ripe bananas, which arise as halos around necrotic areas in 'senescence associated' dark spots. These dark spots appear naturally on the peel of ripe bananas and occur in the vicinity of stomata. Wavelength, space, and time resolved fluorescence measurements allowed the luminescent areas to be monitored on whole bananas. Our studies revealed an accumulation of FCCs in luminescent rings, within senescing cells undergoing the transition to dead tissue, as was observable by morphological textural cellular changes. FCCs typically are short lived intermediates of chlorophyll breakdown. In some plants, FCCs are uniquely persistent, as is seen in bananas, and can thus be used as luminescent in vivo markers in tissue undergoing senescence. While FCCs still remain to be tested for their own hypothetical physiological role in plants, they may help fill the demand for specific endogenous molecular reporters in noninvasive assays of plant senescence. Thus, they allow for in vivo studies, which provide insights into critical stages preceding cell death. PMID- 19805213 TI - Higher trends but larger uncertainty and geographic variability in 21st century temperature and heat waves. AB - Generating credible climate change and extremes projections remains a high priority challenge, especially since recent observed emissions are above the worst-case scenario. Bias and uncertainty analyses of ensemble simulations from a global earth systems model show increased warming and more intense heat waves combined with greater uncertainty and large regional variability in the 21st century. Global warming trends are statistically validated across ensembles and investigated at regional scales. Observed heat wave intensities in the current decade are larger than worst-case projections. Model projections are relatively insensitive to initial conditions, while uncertainty bounds obtained by comparison with recent observations are wider than ensemble ranges. Increased trends in temperature and heat waves, concurrent with larger uncertainty and variability, suggest greater urgency and complexity of adaptation or mitigation decisions. PMID- 19805214 TI - Stability of body-centered cubic iron-magnesium alloys in the Earth's inner core. AB - The composition and the structure of the Earth's solid inner core are still unknown. Iron is accepted to be the main component of the core. Lately, the body centered cubic (bcc) phase of iron was suggested to be present in the inner core, although its stability at core conditions is still in discussion. The higher density of pure iron compared with that of the Earth's core indicates the presence of light element(s) in this region, which could be responsible for the stability of the bcc phase. However, so far, none of the proposed composition models were in full agreement with seismic observations. The solubility of magnesium in hexagonal Fe has been found to increase significantly with increasing pressure, suggesting that Mg can also be an important element in the core. Here, we report a first-principles density functional study of bcc Fe-Mg alloys at core pressures and temperatures. We show that at core conditions, 5-10 atomic percent Mg stabilizes the bcc Fe both dynamically and thermodynamically. Our calculated density, elastic moduli, and sound velocities of bcc Fe-Mg alloys are consistent with those obtained from seismology, indicating that the bcc structured Fe-Mg alloy is a possible model for the Earth's inner core. PMID- 19805215 TI - In vivo far-red luminescence imaging of a biomarker based on BRET from Cypridina bioluminescence to an organic dye. AB - We aimed to develop a far-red luminescence imaging technology for visualization of disease specific antigens on cell surfaces in a living body. First, we conjugated a far-red fluorescent indocyanine derivative to biotinylated Cypridina luciferase. This conjugate produced a bimodal spectrum that has long-wavelength bioluminescence emission in the far-red region as a result of bioluminescence resonance energy transfer. To generate a far-red luminescent probe with targeting and imaging capabilities of tumors, we then linked this conjugate to an anti human Dlk-1 monoclonal antibody via the biotin-avidin interaction. This far-red luminescent probe enabled us to obtain high-resolution microscopic images of live, Dlk-1-expressing Huh-7 cells without an external light source, and to monitor the accumulation of this probe in tumor-bearing mice. Thus this far-red luminescent probe is a convenient analytical tool for the evaluations of monoclonal antibody localization in a living body. PMID- 19805216 TI - Assembly of the Cdc45-Mcm2-7-GINS complex in human cells requires the Ctf4/And-1, RecQL4, and Mcm10 proteins. AB - In eukaryotes, the activation of the prereplicative complex and assembly of an active DNA unwinding complex are critical but poorly understood steps required for the initiation of DNA replication. In this report, we have used bimolecular fluorescence complementation assays in HeLa cells to examine the interactions between Cdc45, Mcm2-7, and the GINS complex (collectively called the CMG complex), which seem to play a key role in the formation and progression of replication forks. Interactions between the CMG components were observed only after the G(1)/S transition of the cell cycle and were abolished by treatment of cells with either a CDK inhibitor or siRNA against the Cdc7 kinase. Stable association of CMG required all three components of the CMG complex as well as RecQL4, Ctf4/And-1, and Mcm10. Surprisingly, depletion of TopBP1, a homologue of Dpb11 that plays an essential role in the chromatin loading of Cdc45 and GINS in yeast cells, did not significantly affect CMG complex formation. These results suggest that the proteins involved in the assembly of initiation complexes in human cells may differ somewhat from those in yeast systems. PMID- 19805217 TI - Identification of an RNA-dependent RNA polymerase in Drosophila involved in RNAi and transposon suppression. AB - Here, we show that recombinant Drosophila elp1 (D-elp1) produced in Sf9 cells or Escherichia coli, corresponding to the largest of the three subunits in the RNA polymerase II core elongator complex, has RNA-dependent RNA polymerase (RdRP) activity. D-elp1 is a noncanonical RdRP that can synthesize dsRNA from different ssRNA templates using either a primer-dependent or primer-independent initiation mechanism. Of the three core subunits, only D-elp1 depletion inhibits RNAi in S2 cells but does not affect micro RNA function. Furthermore, D-elp1 depletion results in increased steady state levels of representative transposon RNAs and a decrease in the corresponding transposon antisense transcripts and endo siRNAs. In contrast, although Dcr-2 depletion results in increased transposon RNA levels and a reduction in the corresponding endo siRNAs, there is no change in the transposon antisense RNA levels. In D-elp1 null third instar larvae transposon RNA levels are also increased and the corresponding transposon antisense RNAs are reduced. D-elp1 associates tightly with Dcr-2, similar to the Dicer-RdRP interaction observed in lower eukaryotes. These results identify an aspect of the RNAi pathway in Drosophila that suggest transposon derived endo siRNAs, critical for transposon suppression, are produced, in part, in a D-elp1 dependent step that converts transposon RNA into dsRNA that is subsequently processed by Dcr-2. The generality of this mechanism in genome defense and RNA silencing in higher eukaryotes is suggested. PMID- 19805218 TI - Protein contents in biological membranes can explain abnormal solvation of charged and polar residues. AB - Transmembrane helices are generally believed to insert into membranes based on their hydrophobicity. Nevertheless, there are important exceptions where polar residues have great functional importance, for instance the S4 helix of voltage gated ion channels. It has been shown experimentally that insertion can be accomplished by hydrophobic counterbalance, predicting an arginine insertion cost of only 2.5 kcal/mol, compared with 14.9 kcal/mol in cyclohexane. Previous simulations of pure bilayers have produced values close to the pure hydrocarbon, which has lead to spirited discussion about the experimental conditions. Here, we have performed computer simulations of models better mimicking biological membranes by explicitly including protein helices at mass fractions from 15% to 55%, as well as an actual translocon. This has a striking effect on the solvation free energy of arginine. With some polar residues present, the solvation cost comes close to experimental observation at approximately 30% mass fraction, and negligible at 40%. In the presence of a translocon in the membrane, the cost of inserting arginine next to the lateral gate can be as low as 3-5 kcal/mol. The effect is mainly due to the extra helices making it easier to retain hydration water. These results offer a possible explanation for the discrepancy between the in vivo hydrophobicity scale and computer simulations and highlight the importance of the high protein contents in membranes. Although many membrane proteins are stable in pure bilayers, such simplified models might not be sufficiently accurate for insertion of polar or charged residues in biological membranes. PMID- 19805219 TI - The continuity of protein structure space is an intrinsic property of proteins. AB - The classical view of the space of protein structures is that it is populated by a discrete set of protein folds. For proteins up to 200 residues long, by using structural alignments and building upon ideas of the completeness and continuity of structure space, we show that nearly any structure is significantly related to any other using a transitive set of no more than 7 intermediate structurally related proteins. This result holds for all structures in the Protein Data Bank, even when structural relationships between evolutionary related proteins (as detected by threading or functional analyses) are excluded. A similar picture holds for an artificial library of compact, hydrogen-bonded, homopolypeptide structures. The 3 sets share the global connectivity features of random graphs, in which the local connectivity of each node (i.e., the number of neighboring structures per protein) is preserved. This high connectivity supports the continuous view of single-domain protein structure space. More importantly, these results do not depend on evolution, rather just on the physics of protein structures. The fact that evolutionary divergence need not be invoked to explain the continuous nature of protein structure space has implications for how the universe of protein structures might have originated, and how function should be transferred between proteins of similar structure. PMID- 19805220 TI - Feeder-free derivation of induced pluripotent stem cells from adult human adipose stem cells. AB - Ectopic expression of transcription factors can reprogram somatic cells to a pluripotent state. However, most of the studies used skin fibroblasts as the starting population for reprogramming, which usually take weeks for expansion from a single biopsy. We show here that induced pluripotent stem (iPS) cells can be generated from adult human adipose stem cells (hASCs) freshly isolated from patients. Furthermore, iPS cells can be readily derived from adult hASCs in a feeder-free condition, thereby eliminating potential variability caused by using feeder cells. hASCs can be safely and readily isolated from adult humans in large quantities without extended time for expansion, are easy to maintain in culture, and therefore represent an ideal autologous source of cells for generating individual-specific iPS cells. PMID- 19805221 TI - Phosphorylation of WASp is a key regulator of activity and stability in vivo. AB - The Wiskott-Aldrich syndrome protein (WASp) is a key cytoskeletal regulator in hematopoietic cells. Covalent modification of a conserved tyrosine by phosphorylation has emerged as an important potential determinant of activity, although the physiological significance remains uncertain. In a murine knockin model, mutation resulting in inability to phosphorylate Y293 (Y293F) mimicked many features of complete WASp-deficiency. Although a phosphomimicking mutant Y293E conferred enhanced actin-polymerization, the cellular phenotype was similar due to functional dysregulation. Furthermore, steady-state levels of Y293E-WASp were markedly reduced compared to wild-type WASp and Y293F-WASp, although partially recoverable by treatment of cells with proteasome inhibitors. Consequently, tyrosine phosphorylation plays a critical role in normal activation of WASp in vivo, and is indispensible for multiple tasks including proliferation, phagocytosis, chemotaxis, and assembly of adhesion structures. Furthermore, it may target WASp for proteasome-mediated degradation, thereby providing a default mechanism for self-limiting stimulation of the Arp2/3 complex. PMID- 19805222 TI - CKIepsilon/delta-dependent phosphorylation is a temperature-insensitive, period determining process in the mammalian circadian clock. AB - A striking feature of the circadian clock is its flexible yet robust response to various environmental conditions. To analyze the biochemical processes underlying this flexible-yet-robust characteristic, we examined the effects of 1,260 pharmacologically active compounds in mouse and human clock cell lines. Compounds that markedly (>10 s.d.) lengthened the period in both cell lines, also lengthened it in central clock tissues and peripheral clock cells. Most compounds inhibited casein kinase Iepsilon (CKIepsilon) or CKIdelta phosphorylation of the PER2 protein. Manipulation of CKIepsilon/delta-dependent phosphorylation by these compounds lengthened the period of the mammalian clock from circadian (24 h) to circabidian (48 h), revealing its high sensitivity to chemical perturbation. The degradation rate of PER2, which is regulated by CKIepsilon/delta-dependent phosphorylation, was temperature-insensitive in living clock cells, yet sensitive to chemical perturbations. This temperature-insensitivity was preserved in the CKIepsilon/delta-dependent phosphorylation of a synthetic peptide in vitro. Thus, CKIepsilon/delta-dependent phosphorylation is likely a temperature-insensitive period-determining process in the mammalian circadian clock. PMID- 19805223 TI - The p53 target Wig-1 regulates p53 mRNA stability through an AU-rich element. AB - The p53 target gene Wig-1 encodes a double-stranded-RNA-binding zinc finger protein. We show here that Wig-1 binds to p53 mRNA and stabilizes it through an AU-rich element (ARE) in the 3' UTR of the p53 mRNA. This effect is mirrored by enhanced p53 protein levels in both unstressed cells and cells exposed to p53 activating stress agents. Thus, the p53 target Wig-1 is a previously undescribed ARE-regulating protein that acts as a positive feedback regulator of p53, with implications both for the steady-state levels of p53 and for the p53 stress response. Our data reveal a previously undescribed link between the tumor suppressor p53 and posttranscriptional gene regulation via AREs in mRNA. PMID- 19805224 TI - The evolution of information suppression in communicating robots with conflicting interests. AB - Reliable information is a crucial factor influencing decision-making and, thus, fitness in all animals. A common source of information comes from inadvertent cues produced by the behavior of conspecifics. Here we use a system of experimental evolution with robots foraging in an arena containing a food source to study how communication strategies can evolve to regulate information provided by such cues. The robots could produce information by emitting blue light, which the other robots could perceive with their cameras. Over the first few generations, the robots quickly evolved to successfully locate the food, while emitting light randomly. This behavior resulted in a high intensity of light near food, which provided social information allowing other robots to more rapidly find the food. Because robots were competing for food, they were quickly selected to conceal this information. However, they never completely ceased to produce information. Detailed analyses revealed that this somewhat surprising result was due to the strength of selection on suppressing information declining concomitantly with the reduction in information content. Accordingly, a stable equilibrium with low information and considerable variation in communicative behaviors was attained by mutation selection. Because a similar coevolutionary process should be common in natural systems, this may explain why communicative strategies are so variable in many animal species. PMID- 19805225 TI - Acp36DE is required for uterine conformational changes in mated Drosophila females. AB - In a multitude of animals with internal fertilization, including insects and mammals, sperm are stored within a female's reproductive tract after mating. Defects in the process of sperm storage drastically reduce reproductive success. In Drosophila males, "Acp" seminal proteins alter female postmating physiology and behavior, and are necessary for several aspects of sperm storage. For example, Acps cause a series of conformational changes in the mated female's reproductive tract that occur during and immediately after mating. These conformational changes have been hypothesized to aid both in the movement of sperm within the female and in the subsequent storage of those sperm. We used RNAi to systematically knock down several Acps involved in sperm storage to determine whether they played a role in the mating-induced uterine conformational changes. Mates of males lacking the glycoprotein Acp36DE, which is needed for the accumulation of sperm in the storage organs, fail to complete the full sequence of the conformational changes. Our results show that uterine conformational changes are important for proper accumulation of sperm in storage and identify a seminal protein that mediates these changes. Four Acps included in this study, previously shown to affect sperm release from storage (CG9997, CG1656, CG1652, and CG17575), are not necessary for uterine conformational changes to occur. Rather, consistent with their role in later steps of sperm storage, we show here that their presence can affect the outcome of sperm competition situations. PMID- 19805226 TI - PD-1+ memory phenotype CD4+ T cells expressing C/EBPalpha underlie T cell immunodepression in senescence and leukemia. AB - Although altered T cell function plays a part in immunosenescence, the mechanisms remain uncertain. Here we identify a bona fide age-dependent PD-1(+) memory phenotype (MP) CD4(+) T cell subpopulation that hardly proliferates in response to T cell receptor (TCR) stimulation and produces abundant osteopontin at the cost of typical T cell lymphokines. These T cells demonstrate impaired repopulation in Rag2(-/-) mice, but a homeostatic proliferation in gamma-ray irradiated mice. These T cells also reveal a unique molecular signature, including a strong expression of C/EBPalpha normally expressed in myeloid-lineage cells, with diminished c-Myc and cyclin D1. Transduction of Cebpa in regular CD4(+) T cells inhibited the TCR-mediated proliferation with c-Myc and cyclin D1 repression and caused a striking activation of Spp1 encoding osteopontin along with concomitant repression of T cell lymphokine genes. Although these T cells gradually increase in number with age and become predominant at the senescent stage in normal mice, the generation is robustly accelerated during leukemia. In both conditions, their predominance is associated with the diminution of specific CD4(+) T cell response. The results suggest that global T cell immunodepression in senescence and leukemia is attributable to the increase in PD-1(+) MP CD4(+) T cells expressing C/EBPalpha. PMID- 19805227 TI - Nod2 is required for the regulation of commensal microbiota in the intestine. AB - Mutations in the Nod2 gene are among the strongest genetic risk factors in the pathogenesis of ileal Crohn's disease, but the exact contributions of Nod2 to intestinal mucosal homeostasis are not understood. Here we show that Nod2 plays an essential role in controlling commensal bacterial flora in the intestine. Analysis of intestinal bacteria from the terminal ilea of Nod2-deficient mice showed that they harbor an increased load of commensal resident bacteria. Furthermore, Nod2-deficient mice had a diminished ability to prevent intestinal colonization of pathogenic bacteria. In vitro, intestinal crypts isolated from terminal ilea of Nod2-deficient mice were unable to kill bacteria effectively, suggesting an important role of Nod2 signaling in crypt function. Interestingly, the expression of Nod2 is dependent on the presence of commensal bacteria, because mice re-derived into germ-free conditions expressed significantly less Nod2 in their terminal ilea, and complementation of commensal bacteria into germ free mice induced Nod2 expression. Therefore, Nod2 and intestinal commensal bacterial flora maintain a balance by regulating each other through a feedback mechanism. Dysfunction of Nod2 results in a break-down of this homeostasis. PMID- 19805228 TI - Antibody-mediated blockade of IL-15 reverses the autoimmune intestinal damage in transgenic mice that overexpress IL-15 in enterocytes. AB - Celiac disease (CD) is an autoimmune inflammatory disease with a relatively high prevalence especially in the western hemisphere. A strong genetic component is involved in the pathogenesis of CD with virtually all individuals that develop the disease carrying HLA-DQ alleles that encode specific HLA-DQ2 or HLA-DQ8 heterodimers. Consumption of cereals rich in gluten triggers a chronic intestinal inflammation in genetically susceptible individuals leading to the development of CD. Emerging evidence has implicated a central role for IL-15 in the orchestration and perpetuation of inflammation and tissue destruction in CD. Therefore, IL-15 represents an attractive target for development of new therapies for CD. Transgenic mice that express human IL-15 specifically in enterocytes (T3(b)-hIL-15 Tg mice) develop villous atrophy and severe duodeno-jejunal inflammation with massive accumulation of NK-like CD8(+) lymphocytes in the affected mucosa. We used these mice to demonstrate that blockade of IL-15 signaling with an antibody (TM-beta1) that binds to murine IL-2/IL-15Rbeta (CD122) leads to a reversal of the autoimmune intestinal damage. The present study, along with work of others, provides the rationale to explore IL-15 blockade as a test of the hypothesis that uncontrolled expression of IL-15 is critical in the pathogenesis and maintenance of refractory CD. PMID- 19805229 TI - DISC1 splice variants are upregulated in schizophrenia and associated with risk polymorphisms. AB - Disrupted-In-Schizophrenia-1 (DISC1) is a promising susceptibility gene for major mental illness, but the mechanism of the clinical association is unknown. We searched for DISC1 transcripts in adult and fetal human brain and tested whether their expression is altered in patients with schizophrenia and is associated with genetic variation in DISC1. Many alternatively spliced transcripts were identified, including groups lacking exon 3 (Delta3), exons 7 and 8 (Delta7Delta8), an exon 3 insertion variant (extra short variant-1, Esv1), and intergenic splicing between TSNAX and DISC1. Isoforms Delta7Delta8, Esv1, and Delta3, which encode truncated DISC1 proteins, were expressed more abundantly during fetal development than during postnatal ages, and their expression was higher in the hippocampus of patients with schizophrenia. Schizophrenia risk associated polymorphisms [non-synonymous SNPs rs821616 (Cys704Ser) and rs6675281 (Leu607Phe), and rs821597] were associated with the expression of Delta3 and Delta7Delta8. Moreover, the same allele at rs6675281, which predicted higher expression of these transcripts in the hippocampus, was associated with higher expression of DISC1Delta7Delta8 in lymphoblasts in an independent sample. Our results implicate a molecular mechanism of genetic risk associated with DISC1 involving specific alterations in gene processing. PMID- 19805230 TI - Rewiring the RNAs of influenza virus to prevent reassortment. AB - Influenza viruses contain segmented, negative-strand RNA genomes. Genome segmentation facilitates reassortment between different influenza virus strains infecting the same cell. This phenomenon results in the rapid exchange of RNA segments. In this study, we have developed a method to prevent the free reassortment of influenza A virus RNAs by rewiring their packaging signals. Specific packaging signals for individual influenza virus RNA segments are located in the 5' and 3' noncoding regions as well as in the terminal regions of the ORF of an RNA segment. By putting the nonstructural protein (NS)-specific packaging sequences onto the ORF of the hemagglutinin (HA) gene and mutating the packaging regions in the ORF of the HA, we created a chimeric HA segment with the packaging identity of an NS gene. By the same strategy, we made an NS gene with the packaging identity of an HA segment. This rewired virus had the packaging signals for all eight influenza virus RNAs, but it lost the ability to independently reassort its HA or NS gene. A similar approach can be applied to the other influenza A virus segments to diminish their ability to form reassortant viruses. PMID- 19805231 TI - Comparative systems biology across an evolutionary gradient within the Shewanella genus. AB - To what extent genotypic differences translate to phenotypic variation remains a poorly understood issue of paramount importance for several cornerstone concepts of microbiology including the species definition. Here, we take advantage of the completed genomic sequences, expressed proteomic profiles, and physiological studies of 10 closely related Shewanella strains and species to provide quantitative insights into this issue. Our analyses revealed that, despite extensive horizontal gene transfer within these genomes, the genotypic and phenotypic similarities among the organisms were generally predictable from their evolutionary relatedness. The power of the predictions depended on the degree of ecological specialization of the organisms evaluated. Using the gradient of evolutionary relatedness formed by these genomes, we were able to partly isolate the effect of ecology from that of evolutionary divergence and to rank the different cellular functions in terms of their rates of evolution. Our ranking also revealed that whole-cell protein expression differences among these organisms, when the organisms were grown under identical conditions, were relatively larger than differences at the genome level, suggesting that similarity in gene regulation and expression should constitute another important parameter for (new) species description. Collectively, our results provide important new information toward beginning a systems-level understanding of bacterial species and genera. PMID- 19805232 TI - Hydrogen is a preferred intermediate in the energy-conserving electron transport chain of Methanosarcina barkeri. AB - Methanogens use an unusual energy-conserving electron transport chain that involves reduction of a limited number of electron acceptors to methane gas. Previous biochemical studies suggested that the proton-pumping F(420)H(2) dehydrogenase (Fpo) plays a crucial role in this process during growth on methanol. However, Methanosarcina barkeri Delta fpo mutants constructed in this study display no measurable phenotype on this substrate, indicating that Fpo plays a minor role, if any. In contrast, Delta frh mutants lacking the cytoplasmic F(420)-reducing hydrogenase (Frh) are severely affected in their ability to grow and make methane from methanol, and double Delta fpo/Delta frh mutants are completely unable to use this substrate. These data suggest that the preferred electron transport chain involves production of hydrogen gas in the cytoplasm, which then diffuses out of the cell, where it is reoxidized with transfer of electrons into the energy-conserving electron transport chain. This hydrogen-cycling metabolism leads directly to production of a proton motive force that can be used by the cell for ATP synthesis. Nevertheless, M. barkeri does have the flexibility to use the Fpo-dependent electron transport chain when needed, as shown by the poor growth of the Delta frh mutant. Our data suggest that the rapid enzymatic turnover of hydrogenases may allow a competitive advantage via faster growth rates in this freshwater organism. The mutant analysis also confirms the proposed role of Frh in growth on hydrogen/carbon dioxide and suggests that either Frh or Fpo is needed for aceticlastic growth of M. barkeri. PMID- 19805233 TI - Functional requirement of AgRP and NPY neurons in ovarian cycle-dependent regulation of food intake. AB - In female mammals including rodents and humans, feeding decreases during the periovulatory period of the ovarian cycle, which coincides with a surge in circulating estrogen levels. Ovariectomy increases food intake, which can be normalized by estrogen treatment at a dose and frequency mimicking those during the estrous cycle. Furthermore, administration of estrogen to rodents potently inhibits food intake. Despite these well-known effects of estrogen, neuronal subtypes that mediate estrogen's anorexigenic effects have not been identified. In this study, we show that changes in hypothalamic expression of agouti-related protein (Agrp) and neuropeptide Y (Npy) coincide with the cyclic changes in feeding across the estrous cycle. These cyclic changes in feeding are abolished in mice with degenerated AgRP neurons even though these mice cycle normally. Central administration of 17beta-estradiol (E2) decreases food intake in controls but not in mice lacking the AgRP neurons. Furthermore, E2 treatment suppresses fasting-induced c-Fos activation in AgRP and NPY neurons and blunts the refeeding response. Surprisingly, although estrogen receptor alpha (ERalpha) is the key mediator of estrogen's anorexigenic effects, we find that expression of ERalpha is completely excluded from AgRP and NPY neurons in the mouse hypothalamus, suggesting that estrogen may regulate these neurons indirectly via presynaptic neurons that express ERalpha. This study indicates that neurons coexpressing AgRP and NPY are functionally required for the cyclic changes in feeding across estrous cycle and that AgRP and NPY neurons are essential mediators of estrogen's anorexigenic function. PMID- 19805234 TI - Reelin signaling antagonizes beta-amyloid at the synapse. AB - Abnormal processing of the amyloid precursor protein (APP) and beta-amyloid (Abeta) plaque accumulation are defining features of Alzheimer disease (AD), a genetically complex neurodegenerative disease that is characterized by progressive synapse loss and neuronal cell death. Abeta induces synaptic dysfunction in part by altering the endocytosis and trafficking of AMPA and NMDA receptors. Reelin is a neuromodulator that increases glutamatergic neurotransmission by signaling through the postsynaptic ApoE receptors Apoer2 and Vldlr and thereby potently enhances synaptic plasticity. Here we show that Reelin can prevent the suppression of long-term potentiation and NMDA receptors, which is induced by levels of Abeta comparable to those present in an AD-afflicted brain. This reversal is dependent upon the activation of Src family tyrosine kinases. At high concentrations of Abeta peptides, Reelin can no longer overcome the Abeta induced functional suppression and this coincides with a complete blockade of the Reelin-dependent phosphorylation of NR2 subunits. We propose a model in which Abeta, Reelin, and ApoE receptors modulate neurotransmission and thus synaptic stability as opposing regulators of synaptic gain control. PMID- 19805235 TI - Reduced arsenic clearance and increased toxicity in aquaglyceroporin-9-null mice. AB - Expressed in liver, aquaglyceroporin-9 (AQP9) is permeated by glycerol, arsenite, and other small, neutral solutes. To evaluate a possible protective role, AQP9 null mice were evaluated for in vivo arsenic toxicity. After injection with NaAsO(2), AQP9-null mice suffer reduced survival rates (LD(50), 12 mg/kg) compared with WT mice (LD(50), 15 mg/kg). The highest tissue level of arsenic is in heart, with AQP9-null mice accumulating 10-20 times more arsenic than WT mice. Within hours after NaAsO(2) injection, AQP9-null mice sustain profound bradycardia, despite normal serum electrolytes. Increased arsenic levels are also present in liver, lung, spleen, and testis of AQP9-null mice. Arsenic levels in the feces and urine of AQP9-null mice are only approximately 10% of the WT levels, and reduced clearance of multiple arsenic species by the AQP9-null mice suggests that AQP9 is involved in the export of multiple forms of arsenic. Immunohistochemical staining of liver sections revealed that AQP9 is most abundant in basolateral membrane of hepatocytes adjacent to the sinusoids. AQP9 is not detected in heart or kidney by PCR or immunohistochemistry. We propose that AQP9 provides a route for excretion of arsenic by the liver, thereby providing partial protection of the whole animal from arsenic toxicity. PMID- 19805236 TI - Secretin as a neurohypophysial factor regulating body water homeostasis. AB - Hypothalamic magnocellular neurons express either one of the neurohypophysial hormones, vasopressin or oxytocin, along with different neuropeptides or neuromodulators. Axonal terminals of these neurons are generally accepted to release solely the two hormones but not others into the circulation. Here, we show that secretin, originally isolated from upper intestinal mucosal extract, is present throughout the hypothalamo-neurohypophysial axis and that it is released from the posterior pituitary under plasma hyperosmolality conditions. In the hypothalamus, it stimulates vasopressin expression and release. Considering these findings together with our previous findings that show a direct effect of secretin on renal water reabsorption, we propose here that secretin works at multiple levels in the hypothalamus, pituitary, and kidney to regulate water homeostasis. Findings presented here challenge previous understanding regarding the neurohypophysis and could provide new concepts in treating disorders related to osmoregulation. PMID- 19805237 TI - Impaired respiration discloses the physiological significance of state transitions in Chlamydomonas. AB - State transitions correspond to a major regulation process for photosynthesis, whereby chlorophyll protein complexes responsible for light harvesting migrate between photosystem II and photosystem I in response to changes in the redox poise of the intersystem electron carriers. Here we disclose their physiological significance in Chlamydomonas reinhardtii using a genetic approach. Using single and double mutants defective for state transitions and/or mitochondrial respiration, we show that photosynthetic growth, and therefore biomass production, critically depends on state transitions in respiratory-defective conditions. When extra ATP cannot be provided by respiration, enhanced photosystem I turnover elicited by transition to state 2 is required for photosynthetic activity. Concomitant impairment of state transitions and respiration decreases the overall yield of photosynthesis, ultimately leading to reduced fitness. We thus provide experimental evidence that the combined energetic contributions of state transitions and respiration are required for efficient carbon assimilation in this alga. PMID- 19805238 TI - Hormonal anticipation of territorial challenges in cichlid fish. AB - In many territorial species androgens respond to social interactions. This response has been interpreted as a mechanism for adjusting aggressive motivation to a changing social environment. Therefore, it would be adaptive to anticipate social challenges and reacting to their clues with an anticipatory androgen response to adjust agonistic motivation to an imminent social challenge. Here we test the hypothesis of an anticipatory androgen response to territorial intrusions using classical conditioning to establish an association between a conditioned stimulus (CS = light) and an unconditioned stimulus (US = intruder male) in male cichlid fish (Oreochromis mossambicus). During the training phase conditioned males (CS-US paired presentations) showed a higher decrease in latency for agonistic response toward the intruder than unconditioned males (CS US unpaired presentations). In the test trial, conditioned males showed an increase in androgen levels (i.e., testosterone and 11-ketotestosterone) relative to baseline, in response to the CS alone. This increase was similar to that of control males exposed to real intruders after CS, whereas unconditioned males showed a decrease in androgen levels in response to the CS. Furthermore, conditioned males were significantly more aggressive than unconditioned males during the post-CS period on test trial, even though the intruder male was not present during this period. These results reveal the occurrence of a conditioned androgen response that may give territorial males an advantage in mounting a defense to upcoming territorial intrusions, if the ability to readily elevate androgens does not co-vary with other traits that bear costs. PMID- 19805239 TI - Coexistence of binocular integration and suppression determined by surface border information. AB - The visual system relies on both the integration and interocular inhibitory processes to achieve single vision from different images in the two eyes. It is generally assumed that the integration process first searches for matching local features between the two eyes. If the matching fails, an interocular inhibitory process is triggered to suppress the image representation of one eye, leading to visual perception that is essentially contributed by the other eye. Here, using a stimulus comprising of binocularly corresponding features (vertical gratings) but incompatible surface border information, we found evidence to the contrary. In one half-image, a circular patch of vertical grating was phase-shifted relative to the surrounding vertical grating to create a circular, monocular boundary contour (MBC), while the other half-image had a similar vertical grating. The two half-images had a binocular disparity at the circular grating patch area, leading to the percept of a disc in depth. Concurrent with the stereo percept, threshold for detecting a Gabor probe on the half-image without the MBC was higher than that on the corresponding area with the grating disc, indicating binocular suppression. These findings reveal that when we perceive depth, which requires the integration process to obtain binocular disparity from the two eyes, one eye's image could simultaneously be suppressed from visual awareness by the interocular inhibitory process. Our study also presents a provocative example of where the brain selectively binds some, but not all, features of the images from the two eyes for visual perception. PMID- 19805240 TI - "Real-time" obstacle avoidance in the absence of primary visual cortex. AB - When we reach toward objects, we easily avoid potential obstacles located in the workspace. Previous studies suggest that obstacle avoidance relies on mechanisms in the dorsal visual stream in the posterior parietal cortex. One fundamental question that remains unanswered is where the visual inputs to these dorsal stream mechanisms are coming from. Here, we provide compelling evidence that these mechanisms can operate in "real-time" without direct input from primary visual cortex (V1). In our first experiment, we used a reaching task to demonstrate that an individual with a dense left visual field hemianopia after damage to V1 remained strikingly sensitive to the position of unseen static obstacles placed in his blind field. Importantly, in a second experiment, we showed that his sensitivity to the same obstacles in his blind field was abolished when a short 2-s delay (without vision) was introduced before reach onset. These findings have far-reaching implications, not only for our understanding of the time constraints under which different visual pathways operate, but also in relation to how these seemingly "primitive" subcortical visual pathways can control complex everyday behavior without recourse to conscious vision. PMID- 19805241 TI - In defense of statistical methods for detecting positive selection. PMID- 19805242 TI - Morphogenera, monophyly, and macroevolution. PMID- 19805243 TI - An improved method for identifying red lakes on art and historical artifacts. PMID- 19805244 TI - The liquid water polymorphism. PMID- 19805245 TI - What is a social tie? PMID- 19805246 TI - The promise and pitfalls of systematic conservation planning. PMID- 19805247 TI - Feeding aquaculture in an era of finite resources. AB - Aquaculture's pressure on forage fisheries remains hotly contested. This article reviews trends in fishmeal and fish oil use in industrial aquafeeds, showing reduced inclusion rates but greater total use associated with increased aquaculture production and demand for fish high in long-chain omega-3 oils. The ratio of wild fisheries inputs to farmed fish output has fallen to 0.63 for the aquaculture sector as a whole but remains as high as 5.0 for Atlantic salmon. Various plant- and animal-based alternatives are now used or available for industrial aquafeeds, depending on relative prices and consumer acceptance, and the outlook for single-cell organisms to replace fish oil is promising. With appropriate economic and regulatory incentives, the transition toward alternative feedstuffs could accelerate, paving the way for a consensus that aquaculture is aiding the ocean, not depleting it. PMID- 19805248 TI - Liquids and structural glasses special feature: liquids: condensed, disordered, and sometimes complex. PMID- 19805249 TI - Poor methodology for predicting large-scale tree die-off. PMID- 19805250 TI - Overlook agricultural research at our peril. PMID- 19805251 TI - Darwin and the recent African origin of modern humans. PMID- 19805252 TI - Finding needles in DNA stacks. PMID- 19805253 TI - ABCs of erythroid mitochondrial iron uptake. PMID- 19805254 TI - Trophoblast stem cells derived from nuclear transfer embryos: phenotypically unique, bad neighbors, or poor communicators? PMID- 19805255 TI - Cytokinin and WUSCHEL tie the knot around plant stem cells. PMID- 19805256 TI - Out of Africa: modern human origins special feature: human origins: out of Africa. AB - Our species, Homo sapiens, is highly autapomorphic (uniquely derived) among hominids in the structure of its skull and postcranial skeleton. It is also sharply distinguished from other organisms by its unique symbolic mode of cognition. The fossil and archaeological records combine to show fairly clearly that our physical and cognitive attributes both first appeared in Africa, but at different times. Essentially modern bony conformation was established in that continent by the 200-150 Ka range (a dating in good agreement with dates for the origin of H. sapiens derived from modern molecular diversity). The event concerned was apparently short-term because it is essentially unanticipated in the fossil record. In contrast, the first convincing stirrings of symbolic behavior are not currently detectable until (possibly well) after 100 Ka. The radical reorganization of gene expression that underwrote the distinctive physical appearance of H. sapiens was probably also responsible for the neural substrate that permits symbolic cognition. This exaptively acquired potential lay unexploited until it was "discovered" via a cultural stimulus, plausibly the invention of language. Modern humans appear to have definitively exited Africa to populate the rest of the globe only after both their physical and cognitive peculiarities had been acquired within that continent. PMID- 19805257 TI - Out of Africa: modern human origins special feature: the origin of Neandertals. AB - Western Eurasia yielded a rich Middle (MP) and Late Pleistocene (LP) fossil record documenting the evolution of the Neandertals that can be analyzed in light of recently acquired paleogenetical data, an abundance of archeological evidence, and a well-known environmental context. Their origin likely relates to an episode of recolonization of Western Eurasia by hominins of African origin carrying the Acheulean technology into Europe around 600 ka. An enhancement of both glacial and interglacial phases may have played a crucial role in this event, as well as in the subsequent evolutionary history of the Western Eurasian populations. In addition to climatic adaptations and an increase in encephalization, genetic drift seems to have played a major role in their evolution. To date, a clear speciation event is not documented, and the most likely scenario for the fixation of Neandertal characteristics seems to be an accretion of features along the second half of the MP. Although a separation time for the African and Eurasian populations is difficult to determine, it certainly predates OIS 11 as phenotypic Neandertal features are documented as far back as and possibly before this time. It is proposed to use the term "Homo rhodesiensis" to designate the large-brained hominins ancestral to H. sapiens in Africa and at the root of the Neandertals in Europe, and to use the term "Homo neanderthalensis" to designate all of the specimens carrying derived metrical or non-metrical features used in the definition of the LP Neandertals. PMID- 19805258 TI - Out of Africa: modern human origins special feature: the meaning of neandertal skeletal morphology. AB - A procedure is outlined for distinguishing among competing hypotheses for fossil morphology and then used to evaluate current views on the meaning of Neandertal skeletal morphology. Three explanations have dominated debates about the meaning of Neandertal cranial features: climatic adaptation, anterior dental loading, and genetic drift. Neither climatic adaptation nor anterior dental loading are well supported, but genetic drift is consistent with the available evidence. Climatic adaptation and activity patterns are the most discussed explanations for Neandertal postcranial features. Robust empirical relationships between climate and body form in extant humans and other endotherms currently make climatic adaptation the most plausible explanation for the wide bodies and relatively short limbs of Neandertals, and many additional postcranial features are likely secondary consequences of these overall skeletal proportions. Activity patterns may explain certain Neandertal postcranial features, but unlike the situation for climate, relationships in extant humans between morphology and activities are typically not well established. For both the cranium and the postcranium, changes in diet or activity patterns may underlie why Neandertals and Pleistocene modern humans tend to be more robust than Holocene humans. PMID- 19805259 TI - Fabrication of large binary colloidal crystals with a NaCl structure. AB - Binary colloidal crystals offer great potential for tuning material properties for applications in, for example, photonics, semiconductors and spintronics, because they allow the positioning of particles with quite different characteristics on one lattice. For micrometer-sized colloids, it is believed that gravity and slow crystallization rates hinder the formation of high-quality binary crystals. Here, we present methods for growing binary colloidal crystals with a NaCl structure from relatively heavy, hard-sphere-like, micrometer-sized silica particles by exploring the following external fields: electric, gravitational, and dielectrophoretic fields and a structured surface (colloidal epitaxy). Our simulations show that the free-energy difference between the NaCl and NiAs structures, which differ in their stacking of the hexagonal planes of the larger spheres, is very small (approximately 0.002 k(B)T). However, we demonstrate that the fcc stacking of the large spheres, which is crucial for obtaining the pure NaCl structure, can be favored by using a combination of the above-mentioned external fields. In this way, we have successfully fabricated large, 3D, oriented single crystals having a NaCl structure without stacking disorder. PMID- 19805260 TI - A simple ligand that selectively targets CUG trinucleotide repeats and inhibits MBNL protein binding. AB - This work describes the rational design, synthesis, and study of a ligand that selectively complexes CUG repeats in RNA (and CTG repeats in DNA) with high nanomolar affinity. This sequence is considered a causative agent of myotonic dystrophy type 1 (DM1) because of its ability to sequester muscleblind-like (MBNL) proteins. Ligand 1 was synthesized in two steps from commercially available compounds, and its binding to CTG and CUG repeats in oligonucleotides studied. Isothermal titration calorimetry studies of 1 with various sequences showed a preference toward the T-T mismatch (K(d) of 390 +/- 80 nM) with a 13-, 169-, and 85-fold reduction in affinity toward single C-C, A-A, and G-G mismatches, respectively. Binding and Job analysis of 1 to multiple CTG step sequences revealed high affinity binding to every other T-T mismatch with negative cooperativity for proximal T-T mismatches. The affinity of 1 for a (CUG)(4) step provided a K(d) of 430 nM with a binding stoichiometry of 1:1. The preference for the U-U in RNA was maintained with a 6-, >143-, and >143-fold reduction in affinity toward single C-C, A-A, and G-G mismatches, respectively. Ligand 1 destabilized the complexes formed between MBNL1N and (CUG)(4) and (CUG)(12) with IC(50) values of 52 +/- 20 microM and 46 +/- 7 microM, respectively, and K(i) values of 6 +/- 2 microM and 7 +/- 1 microM, respectively. These values were only minimally altered by the addition of competitor tRNA. Ligand 1 does not destabilize the unrelated RNA-protein complexes the U1A-SL2 RNA complex and the Sex lethal-tra RNA complex. Thus, ligand 1 selectively destabilizes the MBNL1N-poly(CUG) complex. PMID- 19805261 TI - Atypical mechanism of conduction in potassium channels. AB - Potassium channels can conduct passively K+ ions with rates of up to approximately 10(8) ions per second at physiological conditions, and they are selective to these species by a factor of 10(4) over Na+ ions. Ion conduction has been proposed to involve transitions between 2 main states, with 2 or 3 K+ ions occupying the selectivity filter separated by an intervening water molecule. The largest free energy barrier of such a process was reported to be of the order of 2-3 kcal mol(-1). Here, we present an alternative mechanism for conduction of K+ in potassium channels where site vacancies are involved, and we propose that coexistence of several ion permeation mechanisms is energetically possible. Conduction can be described as a more anarchic phenomenon than previously characterized by the concerted translocations of K+-water-K+. PMID- 19805262 TI - Chemical dynamics of triacetylene formation and implications to the synthesis of polyynes in Titan's atmosphere. AB - For the last four decades, the role of polyynes such as diacetylene (HCCCCH) and triacetylene (HCCCCCCH) in the chemical evolution of the atmosphere of Saturn's moon Titan has been a subject of vigorous research. These polyacetylenes are thought to serve as an UV radiation shield in planetary environments; thus, acting as prebiotic ozone, and are considered as important constituents of the visible haze layers on Titan. However, the underlying chemical processes that initiate the formation and control the growth of polyynes have been the least understood to date. Here, we present a combined experimental, theoretical, and modeling study on the synthesis of the polyyne triacetylene (HCCCCCCH) via the bimolecular gas phase reaction of the ethynyl radical (CCH) with diacetylene (HCCCCH). This elementary reaction is rapid, has no entrance barrier, and yields the triacetylene molecule via indirect scattering dynamics through complex formation in a single collision event. Photochemical models of Titan's atmosphere imply that triacetylene may serve as a building block to synthesize even more complex polyynes such as tetraacetylene (HCCCCCCCCH). PMID- 19805263 TI - Spectroscopic evidence for an engineered, catalytically active Trp radical that creates the unique reactivity of lignin peroxidase. AB - The surface oxidation site (Trp-171) in lignin peroxidase (LiP) required for the reaction with veratryl alcohol a high-redox-potential (1.4 V) substrate, was engineered into Coprinus cinereus peroxidase (CiP) by introducing a Trp residue into a heme peroxidase that has similar protein fold but lacks this activity. To create the catalytic activity toward veratryl alcohol in CiP, it was necessary to reproduce the Trp site and its negatively charged microenvironment by means of a triple mutation. The resulting D179W+R258E+R272D variant was characterized by multifrequency EPR spectroscopy. The spectra unequivocally showed that a new Trp radical [g values of g(x) = 2.0035(5), g(y) = 2.0027(5), and g(z) = 2.0022(1)] was formed after the [Fe(IV)=O Por(*+)] intermediate, as a result of intramolecular electron transfer between Trp-179 and the porphyrin. Also, the EPR characterization crucially showed that [Fe(IV)=O Trp-179(*)] was the reactive intermediate with veratryl alcohol. Accordingly, our work shows that it is necessary to take into account the physicochemical properties of the radical, fine-tuned by the microenvironment, as well as those of the preceding [Fe(IV)=O Por(*+)] intermediate to engineer a catalytically competent Trp site for a given substrate. Manipulation of the microenvironment of the Trp-171 site in LiP allowed the detection by EPR spectroscopy of the Trp-171(*), for which direct evidence has been missing so far. Our work also highlights the role of Trp residues as tunable redox-active cofactors for enzyme catalysis in the context of peroxidases with a unique reactivity toward recalcitrant substrates that require oxidation potentials not realized at the heme site. PMID- 19805264 TI - Chemoenzymatic synthesis of GDP-L-fucose and the Lewis X glycan derivatives. AB - Lewis X (Le(x))-containing glycans play important roles in numerous cellular processes. However, the absence of robust, facile, and cost-effective methods for the synthesis of Le(x) and its structurally related analogs has severely hampered the elucidation of the specific functions of these glycan epitopes. Here we demonstrate that chemically defined guanidine 5'-diphosphate-beta-l-fucose (GDP fucose), the universal fucosyl donor, the Le(x) trisaccharide, and their C-5 substituted derivatives can be synthesized on preparative scales, using a chemoenzymatic approach. This method exploits l-fucokinase/GDP-fucose pyrophosphorylase (FKP), a bifunctional enzyme isolated from Bacteroides fragilis 9343, which converts l-fucose into GDP-fucose via a fucose-1-phosphate (Fuc-1-P) intermediate. Combining the activities of FKP and a Helicobacter pylori alpha1,3 fucosyltransferase, we prepared a library of Le(x) trisaccharide glycans bearing a wide variety of functional groups at the fucose C-5 position. These neoglycoconjugates will be invaluable tools for studying Le(x)-mediated biological processes. PMID- 19805265 TI - A realistic molecular model of cement hydrates. AB - Despite decades of studies of calcium-silicate-hydrate (C-S-H), the structurally complex binder phase of concrete, the interplay between chemical composition and density remains essentially unexplored. Together these characteristics of C-S-H define and modulate the physical and mechanical properties of this "liquid stone" gel phase. With the recent determination of the calcium/silicon (C/S = 1.7) ratio and the density of the C-S-H particle (2.6 g/cm(3)) by neutron scattering measurements, there is new urgency to the challenge of explaining these essential properties. Here we propose a molecular model of C-S-H based on a bottom-up atomistic simulation approach that considers only the chemical specificity of the system as the overriding constraint. By allowing for short silica chains distributed as monomers, dimers, and pentamers, this C-S-H archetype of a molecular description of interacting CaO, SiO2, and H2O units provides not only realistic values of the C/S ratio and the density computed by grand canonical Monte Carlo simulation of water adsorption at 300 K. The model, with a chemical composition of (CaO)(1.65)(SiO2)(H2O)(1.75), also predicts other essential structural features and fundamental physical properties amenable to experimental validation, which suggest that the C-S-H gel structure includes both glass-like short-range order and crystalline features of the mineral tobermorite. Additionally, we probe the mechanical stiffness, strength, and hydrolytic shear response of our molecular model, as compared to experimentally measured properties of C-S-H. The latter results illustrate the prospect of treating cement on equal footing with metals and ceramics in the current application of mechanism-based models and multiscale simulations to study inelastic deformation and cracking. PMID- 19805266 TI - Competing grain-boundary- and dislocation-mediated mechanisms in plastic strain recovery in nanocrystalline aluminum. AB - Recent experiments have demonstrated that plastic strains in nanocrystalline aluminum and gold films with grain sizes on the order of 50 nm are partially recoverable. To reveal the mechanisms behind such strain recovery, we perform large scale molecular dynamics simulations of plastic deformation in nanocrystalline aluminum with mean grain sizes of 10, 20, and 30 nm. Our results indicate that the inhomogeneous deformation in a polycrystalline environment results in significant residual stresses in the nanocrystals. Upon unloading, these internal residual stresses cause strain recovery via competitive deformation mechanisms including dislocation reverse motion/annihilation and grain-boundary sliding/diffusion. By tracking the evolution of each individual deformation mechanism during strain recovery, we quantify the fractional contributions by grain-boundary and dislocation deformation mechanisms to the overall recovered strain. Our analysis shows that, even under strain rates as high as those in molecular dynamics simulations, grain-boundary-mediated processes play important roles in the deformation of nanocrystalline aluminum. PMID- 19805267 TI - Polar firn air reveals large-scale impact of anthropogenic mercury emissions during the 1970s. AB - Mercury (Hg) is an extremely toxic pollutant, and its biogeochemical cycle has been perturbed by anthropogenic emissions during recent centuries. In the atmosphere, gaseous elemental mercury (GEM; Hg degrees ) is the predominant form of mercury (up to 95%). Here we report the evolution of atmospheric levels of GEM in mid- to high-northern latitudes inferred from the interstitial air of firn (perennial snowpack) at Summit, Greenland. GEM concentrations increased rapidly after World War II from approximately 1.5 ng m(-3) reaching a maximum of approximately 3 ng m(-3) around 1970 and decreased until stabilizing at approximately 1.7 ng m(-3) around 1995. This reconstruction reproduces real-time measurements available from the Arctic since 1995 and exhibits the same general trend observed in Europe since 1990. Anthropogenic emissions caused a two-fold rise in boreal atmospheric GEM concentrations before the 1970s, which likely contributed to higher deposition of mercury in both industrialized and remotes areas. Once deposited, this toxin becomes available for methylation and, subsequently, the contamination of ecosystems. Implementation of air pollution regulations, however, enabled a large-scale decline in atmospheric mercury levels during the 1980s. The results shown here suggest that potential increases in emissions in the coming decades could have a similar large-scale impact on atmospheric Hg levels. PMID- 19805268 TI - Long-term natural variability and 20th century climate change. AB - Global mean temperature at the Earth's surface responds both to externally imposed forcings, such as those arising from anthropogenic greenhouse gases, as well as to natural modes of variability internal to the climate system. Variability associated with these latter processes, generally referred to as natural long-term climate variability, arises primarily from changes in oceanic circulation. Here we present a technique that objectively identifies the component of inter-decadal global mean surface temperature attributable to natural long-term climate variability. Removal of that hidden variability from the actual observed global mean surface temperature record delineates the externally forced climate signal, which is monotonic, accelerating warming during the 20th century. PMID- 19805269 TI - Quantification of rapid environmental redox processes with quick-scanning x-ray absorption spectroscopy (Q-XAS). AB - Quantification of the initial rates of environmental reactions at the mineral/water interface is a fundamental prerequisite to determining reaction mechanisms and contaminant transport modeling and predicting environmental risk. Until recently, experimental techniques with adequate time resolution and elemental sensitivity to measure initial rates of the wide variety of environmental reactions were quite limited. Techniques such as electron paramagnetic resonance and Fourier transform infrared spectroscopies suffer from limited elemental specificity and poor sensitivity to inorganic elements, respectively. Ex situ analysis of batch and stirred-flow systems provides high elemental sensitivity; however, their time resolution is inadequate to characterize rapid environmental reactions. Here we apply quick-scanning x-ray absorption spectroscopy (Q-XAS), at sub-second time-scales, to measure the initial oxidation rate of As(III) to As(V) by hydrous manganese(IV) oxide. Using Q-XAS, As(III) and As(V) concentrations were determined every 0.98 s in batch reactions. The initial apparent As(III) depletion rate constants (t < 30 s) measured with Q-XAS are nearly twice as large as rate constants measured with traditional analytical techniques. Our results demonstrate the importance of developing analytical techniques capable of analyzing environmental reactions on the same time scale as they occur. Given the high sensitivity, elemental specificity, and time resolution of Q-XAS, it has many potential applications. They could include measuring not only redox reactions but also dissolution/precipitation reactions, such as the formation and/or reductive dissolution of Fe(III) (hydr)oxides, solid-phase transformations (i.e., formation of layered-double hydroxide minerals), or almost any other reaction occurring in aqueous media that can be measured using x-ray absorption spectroscopy. PMID- 19805270 TI - Forest transition in Vietnam and displacement of deforestation abroad. AB - In some countries across the globe, tropical forest cover is increasing. The national-scale reforestation of Vietnam since 1992 is assumed to contribute to this recovery. It is achieved, however, by the displacement of forest extraction to other countries on the order of 49 (34-70) M m(3), or approximately 39% of the regrowth of Vietnam's forests from 1987 to 2006. Approximately half of wood imports to Vietnam during this period were illegal. Leakage due to policies restricting forest exploitation and displacement due to growing domestic consumption and exports contributed respectively to an estimated 58% and 42% of total displacement. Exports of wood products from Vietnam also grew rapidly, amounting to 84% of the displacement, which is a remarkable feature of the forest transition in Vietnam. Attribution of the displacement and corresponding forest extraction to Vietnam, the source countries or the final consumers is thus debatable. Sixty-one percent of the regrowth in Vietnam was, thus, not associated with displacement abroad. Policies allocating credits to countries for reducing deforestation and forest degradation should monitor illegal timber trade and take into account the policy-induced leakage of wood extraction to other countries. PMID- 19805271 TI - Visualizing sodium dynamics in isolated cardiomyocytes using fluorescent nanosensors. AB - Regulation of sodium flux across the cell membrane plays a vital role in the generation of action potentials and regulation of membrane excitability in cells such as cardiomyocytes and neurons. Alteration of sodium channel function has been implicated in diseases such as epilepsy, long QT syndrome, and heart failure. However, single cell imaging of sodium dynamics has been limited due to the narrow selection of fluorescent sodium indicators available to researchers. Here we report, the detection of spatially defined sodium activity during action potentials. Fluorescent nanosensors that measure sodium in real-time, are reversible and are completely selective over other cations such as potassium that were used to image sodium. The use of the nanosensors in vitro was validated by determining drug-induced activation in heterologous cells transfected with the voltage-gated sodium channel Na(V)1.7. Spatial information of sodium concentrations during action potentials will provide insight at the cellular level on the role of sodium and how slight changes in sodium channel function can affect the entirety of an action potential. PMID- 19805272 TI - Tracking the roots of cellulase hyperproduction by the fungus Trichoderma reesei using massively parallel DNA sequencing. AB - Trichoderma reesei (teleomorph Hypocrea jecorina) is the main industrial source of cellulases and hemicellulases harnessed for the hydrolysis of biomass to simple sugars, which can then be converted to biofuels such as ethanol and other chemicals. The highly productive strains in use today were generated by classical mutagenesis. To learn how cellulase production was improved by these techniques, we performed massively parallel sequencing to identify mutations in the genomes of two hyperproducing strains (NG14, and its direct improved descendant, RUT C30). We detected a surprisingly high number of mutagenic events: 223 single nucleotides variants, 15 small deletions or insertions, and 18 larger deletions, leading to the loss of more than 100 kb of genomic DNA. From these events, we report previously undocumented non-synonymous mutations in 43 genes that are mainly involved in nuclear transport, mRNA stability, transcription, secretion/vacuolar targeting, and metabolism. This homogeneity of functional categories suggests that multiple changes are necessary to improve cellulase production and not simply a few clear-cut mutagenic events. Phenotype microarrays show that some of these mutations result in strong changes in the carbon assimilation pattern of the two mutants with respect to the wild-type strain QM6a. Our analysis provides genome-wide insights into the changes induced by classical mutagenesis in a filamentous fungus and suggests areas for the generation of enhanced T. reesei strains for industrial applications such as biofuel production. PMID- 19805273 TI - C-end rule peptides mediate neuropilin-1-dependent cell, vascular, and tissue penetration. AB - Screening of phage libraries expressing random peptides for binding to prostate cancer cells primarily yielded peptides that had a C-terminal arginine (or rarely lysine) residue, usually in a consensus context R/KXXR/K. Phage expressing these sequences and synthetic nanoparticles coated with them bound to and were internalized into cells. The C-terminal arginine (or lysine) was essential to the activity; adding another amino acid, or even blocking the free carboxyl group of this arginine residue by amidation, eliminated the binding and internalizing activity. An internal R/KXXR/K can be exposed and switched on by a cleavage by a protease. The strict requirement for C-terminal exposure of the motif prompted us to term the phenomenon the C-end rule (CendR). Affinity chromatography showed that the CendR peptides bind to neuropilin-1 (NRP-1) on the target cells. NRP-1 is a cell-surface receptor that plays an essential role in angiogenesis, regulation of vascular permeability, and the development of the nervous system. VEGF-A165 and other ligands of NRP-1 possess a C-terminal CendR sequence that interacts with the b1 domain of NRP-1 and causes cellular internalization and vascular leakage. Our CendR peptides have similar effects, particularly when made multivalent through coupling to a particle. We also noted a unique and important activity of these peptides: penetration and transportation through tissues. The peptides were able to take payloads up to the nanoparticle size scale deep into extravascular tissue. Our observations have implications in drug delivery and penetration of tissue barriers and tumors. PMID- 19805274 TI - Profiling protein thiol oxidation in tumor cells using sulfenic acid-specific antibodies. AB - Hydrogen peroxide (H2O2) functions as a second messenger that can activate cell proliferation through chemoselective oxidation of cysteine residues in signaling proteins. The connection between H2O2 signaling, thiol oxidation, and activation of growth pathways has emerged as fertile ground for the development of strategies for cancer treatment. Central to achieving this goal is the development of tools and assays that facilitate characterization of the molecular events associated with tumorigenesis and evaluation of patient response to therapy. Here we report on the development of an immunochemical method for detecting sulfenic acid, the initial oxidation product that results when a thiolate reacts with H2O2. For this approach, the sulfenic acid is derivatized with a chemical tag to generate a unique epitope for recognition. The elicited antibody is exquisitely specific, context-independent, and capable of visualizing sulfenic acid formation in cells. Applying this approach to several systems, including cancer cell lines, shows it can be used to monitor differences in thiol redox status and reveals a diverse pattern of sulfenic acid modifications across different subtypes of breast tumors. These studies demonstrate a general strategy for producing antibodies against a specific oxidation state of cysteine and show the utility of these reagents for profiling thiol oxidation associated with pathological conditions such as breast cancer. PMID- 19805275 TI - The cytochrome ba3 oxygen reductase from Thermus thermophilus uses a single input channel for proton delivery to the active site and for proton pumping. AB - The heme-copper oxygen reductases are redox-driven proton pumps that generate a proton motive force in both prokaryotes and mitochondria. These enzymes have been divided into 3 evolutionarily related groups: the A-, B- and C-families. Most experimental work on proton-pumping mechanisms has been performed with members of the A-family. These enzymes require 2 proton input pathways (D- and K-channels) to transfer protons used for oxygen reduction chemistry and for proton pumping, with the D-channel transporting all pumped protons. In this work we use site directed mutagenesis to demonstrate that the ba(3) oxygen reductase from Thermus thermophilus, a representative of the B-family, does not contain a D-channel. Rather, it utilizes only 1 proton input channel, analogous to that of the A family K-channel, and it delivers protons to the active site for both O2 chemistry and proton pumping. Comparison of available subunit I sequences reveals that the only structural elements conserved within the oxygen reductase families that could perform these functions are active-site components, namely the covalently linked histidine-tyrosine, the Cu(B) and its ligands, and the active site heme and its ligands. Therefore, our data suggest that all oxygen reductases perform the same chemical reactions for oxygen reduction and comprise the essential elements of the proton-pumping mechanism (e.g., the proton-loading and kinetic-gating sites). These sites, however, cannot be located within the D channel. These results along with structural considerations point to the A propionate region of the active-site heme and surrounding water molecules as the proton-loading site. PMID- 19805276 TI - Intramembrane proteolytic cleavage of a membrane-tethered transcription factor by a metalloprotease depends on ATP. AB - Regulated intramembrane proteolysis (RIP) involves cleavage of a transmembrane segment of a protein. RIP governs diverse processes in a wide variety of organisms and is carried out by different types of intramembrane proteases (IPs), including a large family of metalloproteases. The Bacillus subtilis SpoIVFB protein is a putative metalloprotease that cleaves membrane-tethered Pro sigma(K), releasing sigma(K) to direct transcription of genes necessary for spore formation. By attaching an extra transmembrane segment to the N terminus of SpoIVFB, expression in E. coli was improved more than 100-fold, facilitating purification and demonstration of metalloprotease activity, which accurately cleaved purified Pro-sigma(K). Uniquely for IPs examined so far, SpoIVFB activity requires ATP, which binds to the C-terminal cystathionine-beta-synthase (CBS) domain of SpoIVFB. Deleting just 10 residues from the C-terminal end of SpoIVFB nearly eliminated cleavage of coexpressed Pro-sigma(K) in E. coli. The CBS domain of SpoIVFB was shown to interact with Pro-sigma(K) and ATP changed the interaction, suggesting that ATP regulates substrate access to the active site and renders cleavage sensitive to the cellular energy level, which may be a general feature of CBS-domain-containing IPs. Incorporation of SpoIVFB into preformed liposomes stimulated its ability to cleave Pro-sigma(K). Cleavage depended on ATP and the correct peptide bond was shown to be cleaved using a rapid and sensitive mass spectrometry assay. A system for biochemical studies of RIP by a metalloprotease in a membrane environment has been established using methods that might be applicable to other IPs. PMID- 19805277 TI - Crystallographic snapshots of iterative substrate translocations during nicotianamine synthesis in Archaea. AB - Nicotianamine (NA), a small molecule ubiquitous in plants, is an important divalent metal chelator and the main precursor of phytosiderophores. Nicotianamine synthase (NAS) is the enzyme catalyzing NA synthesis by the condensation of three aminopropyl moieties of S-adenosylmethionine (SAM) and the cyclization of one of them to form an azetidine ring. Here we report five crystal structures of an archaeal NAS from Methanothermobacter thermautotrophicus, either free or in complex with its product(s) and substrate(s). These structures reveal a two-domains fold arrangement of MtNAS, a small molecule related to NA (named here thermoNicotianamine or tNA), and an original mechanism of synthesis in a buried reaction chamber. This reaction chamber is open to the solvent through a small inlet, and a single active site allows the selective entrance of only one substrate at a time that is then processed and translocated stepwise. PMID- 19805278 TI - Structural plasticity and catalysis regulation of a thermosensor histidine kinase. AB - Temperature sensing is essential for the survival of living cells. A major challenge is to understand how a biological thermometer processes thermal information to optimize cellular functions. Using structural and biochemical approaches, we show that the thermosensitive histidine kinase, DesK, from Bacillus subtilis is cold-activated through specific interhelical rearrangements in its central four-helix bundle domain. As revealed by the crystal structures of DesK in different functional states, the plasticity of this helical domain influences the catalytic activities of the protein, either by modifying the mobility of the ATP-binding domains for autokinase activity or by modulating binding of the cognate response regulator to sustain the phosphotransferase and phosphatase activities. The structural and biochemical data suggest a model in which the transmembrane sensor domain of DesK promotes these structural changes through conformational signals transmitted by the membrane-connecting two-helical coiled-coil, ultimately controlling the alternation between output autokinase and phosphatase activities. The structural comparison of the different DesK variants indicates that incoming signals can take the form of helix rotations and asymmetric helical bends similar to those reported for other sensing systems, suggesting that a similar switching mechanism could be operational in a wide range of sensor histidine kinases. PMID- 19805279 TI - Phosphoinositides and SNARE chaperones synergistically assemble and remodel SNARE complexes for membrane fusion. AB - Yeast vacuole fusion requires 4 SNAREs, 2 SNARE chaperone systems (Sec17p/Sec18p/ATP and the HOPS complex), and 2 phosphoinositides, phosphatidylinositol 3-phosphate [PI(3)P] and phosphatidylinositol 4,5 bisphosphate [PI(4,5)P(2)]. By reconstituting proteoliposomal fusion with purified components, we now show that phosphoinositides have 4 distinct roles: PI(3)P is recognized by the PX domain of the SNARE Vam7p; PI(3)P enhances the capacity of membrane-bound SNAREs to drive fusion in the absence of SNARE chaperones; either PI(3)P or PI(4,5)P(2) can activate SNARE chaperones for the recruitment of Vam7p into fusion-competent SNARE complexes; and either PI(3)P or PI(4,5)P(2) strikingly promotes synergistic SNARE complex remodeling by Sec17p/Sec18p/ATP and HOPS. This ternary synergy of phosphoinositides and 2 SNARE chaperone systems is required for rapid fusion. PMID- 19805280 TI - An Armadillo motif in Ufd3 interacts with Cdc48 and is involved in ubiquitin homeostasis and protein degradation. AB - The yeast AAA-ATPase Cdc48 and the ubiquitin fusion degradation (UFD) proteins play important, evolutionarily conserved roles in ubiquitin dependent protein degradation. The N-terminal domain of Cdc48 interacts with substrate-recruiting cofactors, whereas the C terminus of Cdc48 binds to proteins such as Ufd3 that process substrates. Ufd3 is essential for efficient protein degradation and for maintaining cellular ubiquitin levels. This protein contains an N-terminal WD40 domain, a central ubiquitin-binding domain, and a C-terminal Cdc48-binding PUL domain. The crystal structure of the PUL domain reveals an Armadillo repeat with high structural similarity to importin-alpha, and the Cdc48-binding site could be mapped to the concave surface of the PUL domain by biochemical studies. Alterations of the Cdc48 binding site of Ufd3 by site-directed mutagenesis resulted in a depletion of cellular ubiquitin pools and reduced activity of the ubiquitin fusion degradation pathway. Therefore, our data provide direct evidence that the functions of Ufd3 in ubiquitin homeostasis and protein degradation depend on its interaction with the C terminus of Cdc48. PMID- 19805281 TI - Limited terminal transferase in human DNA polymerase mu defines the required balance between accuracy and efficiency in NHEJ. AB - DNA polymerase mu (Polmu) is a family X member implicated in DNA repair, with template-directed and terminal transferase (template-independent) activities. It has been proposed that the terminal transferase activity of Polmu can be specifically required during non-homologous end joining (NHEJ) to create or increase complementarity of DNA ends. By site-directed mutagenesis in human Polmu, we have identified a specific DNA ligand residue (Arg(387)) that is responsible for its limited terminal transferase activity compared to that of human TdT, its closest homologue (42% amino acid identity). Polmu mutant R387K (mimicking TdT) displayed a large increase in terminal transferase activity, but a weakened interaction with ssDNA. That paradox can be explained by the regulatory role of Arg(387) in the translocation of the primer from a non productive E:DNA complex to a productive E:DNA:dNTP complex in the absence of a templating base, which is probably the rate limiting step during template independent synthesis. Further, we show that the Polmu switch from terminal transferase to templated insertions in NHEJ reactions is triggered by recognition of a 5'-P at a second DNA end, whose 3'-protrusion could provide a templating base to facilitate such a special "pre-catalytic translocation step." These studies shed light on the mechanism by which a rate-limited terminal transferase activity in Polmu could regulate the balance between accuracy and necessary efficiency, providing some variability during NHEJ. PMID- 19805282 TI - Biogenesis of glutaminyl-mt tRNAGln in human mitochondria. AB - Mammalian mitochondrial (mt) tRNAs, which are required for mitochondrial protein synthesis, are all encoded in the mitochondrial genome, while mt aminoacyl-tRNA synthetases (aaRSs) are encoded in the nuclear genome. However, no mitochondrial homolog of glutaminyl-tRNA synthetase (GlnRS) has been identified in mammalian genomes, implying that Gln-tRNA(Gln) is synthesized via an indirect pathway in the mammalian mitochondria. We demonstrate here that human mt glutamyl-tRNA synthetase (mtGluRS) efficiently misaminoacylates mt tRNA(Gln) to form Glu tRNA(Gln). In addition, we have identified a human homolog of the Glu-tRNA(Gln) amidotransferase, the hGatCAB heterotrimer. When any of the hGatCAB subunits were inactivated by siRNA-mediated knock down in human cells, the Glu-charged form of tRNA(Gln) accumulated and defects in respiration could be observed. We successfully reconstituted in vitro Gln-tRNA(Gln) formation catalyzed by the recombinant mtGluRS and hGatCAB. The misaminoacylated form of tRNA(Gln) has a weak binding affinity to the mt elongation factor Tu (mtEF-Tu), indicating that the misaminoacylated form of tRNA(Gln) is rejected from the translational apparatus to maintain the accuracy of mitochondrial protein synthesis. PMID- 19805283 TI - Structure of a tRNA-dependent kinase essential for selenocysteine decoding. AB - Compared to bacteria, archaea and eukaryotes employ an additional enzyme for the biosynthesis of selenocysteine (Sec), the 21(st) natural amino acid (aa). An essential RNA-dependent kinase, O-phosphoseryl-tRNA(Sec) kinase (PSTK), converts seryl-tRNA(Sec) to O-phosphoseryl-tRNA(Sec), the immediate precursor of selenocysteinyl-tRNA(Sec). The sequence of Methanocaldococcus jannaschii PSTK (MjPSTK) suggests an N-terminal kinase domain (177 aa) followed by a presumed tRNA binding region (75 aa). The structures of MjPSTK complexed with ADP and AMPPNP revealed that this enzyme belongs to the P-loop kinase class, and that the kinase domain is closely related to gluconate kinase and adenylate kinase. ATP is bound by the P-loop domain (residues 11-18). Formed by antiparallel dimerization of two PSTK monomers, the enzyme structure shows a deep groove with positive electrostatic potential. Located in this groove is the enzyme's active site, which biochemical and genetic data suggest is composed of Asp-41, Arg-44, Glu-55, Tyr-82, Tyr-83, Met-86, and Met-132. Based on structural comparison with Escherichia coli adenylate kinase a docking model was generated that assigns these amino acids to the recognition of the terminal A76-Ser moieties of Ser tRNA(Sec). The geometry and electrostatic environment of the groove in MjPSTK are perfectly complementary to the unusually long acceptor helix of tRNA(Sec). PMID- 19805284 TI - NO formation by a catalytically self-sufficient bacterial nitric oxide synthase from Sorangium cellulosum. AB - The role of nitric oxide (NO) in the host response to infection and in cellular signaling is well established. Enzymatic synthesis of NO is catalyzed by the nitric oxide synthases (NOSs), which convert Arg into NO and citrulline using co substrates O2 and NADPH. Mammalian NOS contains a flavin reductase domain (FAD and FMN) and a catalytic heme oxygenase domain (P450-type heme and tetrahydrobiopterin). Bacterial NOSs, while much less studied, were previously identified as only containing the heme oxygenase domain of the more complex mammalian NOSs. We report here on the characterization of a NOS from Sorangium cellulosum (both full-length, scNOS, and oxygenase domain, scNOSox). scNOS contains a catalytic, oxygenase domain similar to those found in the mammalian NOS and in other bacteria. Unlike the other bacterial NOSs reported to date, however, this protein contains a fused reductase domain. The scNOS reductase domain is unique for the entire NOS family because it utilizes a 2Fe2S cluster for electron transfer. scNOS catalytically produces NO and citrulline in the presence of either tetrahydrobiopterin or tetrahydrofolate. These results establish a bacterial electron transfer pathway used for biological NO synthesis as well as a unique flexibility in using different tetrahydropterin cofactors for this reaction. PMID- 19805285 TI - Short-distance probes for protein backbone structure based on energy transfer between bimane and transition metal ions. AB - The structure and dynamics of proteins underlies the workings of virtually every biological process. Existing biophysical methods are inadequate to measure protein structure at atomic resolution, on a rapid time scale, with limited amounts of protein, and in the context of a cell or membrane. FRET can measure distances between two probes, but depends on the orientation of the probes and typically works only over long distances comparable with the size of many proteins. Also, common probes used for FRET can be large and have long, flexible attachment linkers that position dyes far from the protein backbone. Here, we improve and extend a fluorescence method called transition metal ion FRET that uses energy transfer to transition metal ions as a reporter of short-range distances in proteins with little orientation dependence. This method uses a very small cysteine-reactive dye monobromobimane, with virtually no linker, and various transition metal ions bound close to the peptide backbone as the acceptor. We show that, unlike larger fluorophores and longer linkers, this donor acceptor pair accurately reports short-range distances and changes in backbone distances. We further extend the method by using cysteine-reactive metal chelators, which allow the technique to be used in protein regions of unknown secondary structure or when native metal ion binding sites are present. This improved method overcomes several of the key limitations of classical FRET for intramolecular distance measurements. PMID- 19805286 TI - Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. AB - Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the alpha-CA enzyme family, which is involved in solid tumor acidification. It is a marker of tumor hypoxia and a prognostic factor in several human cancers. An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. The structure reveals a typical alpha-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO(2) hydration to bicarbonate and protons. On the basis of the structural differences observed between CA IX and the other membrane-associated alpha-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo. PMID- 19805287 TI - Direct observation of barrier-limited folding of BBL by single-molecule fluorescence resonance energy transfer. AB - One controversial area in protein folding mechanisms is whether some small, ultra fast-folding proteins exist in distinct native and denatured state ensembles, separated by an energy barrier, or if there is a continuum of states between native and denatured. In theory, the simplest way of distinguishing between single-state barrierless or "downhill" folding and conventional separate state folding is by single-molecule spectroscopy, which can detect either distinct populations of proteins or a continuum. But, the time resolution of approximately 1 ms of most confocal fluorescence microscopes for single-molecule fluorescence resonance energy transfer (SM-FRET) is longer than that for the structural relaxation of proteins such as BBL, whose mechanism of folding is controversial. We have constructed a highly sensitive confocal fluorescence microscope and measured the distribution of FRET efficiencies of appropriately labeled BBL in time bins of 50 and 200 mus under conditions in which its structural relaxation time is 340 mus or less. The experiments are at the very limits of detection because of signal artefacts from shot noise, photo-bleaching, and other events that broaden signals of individual states so they appear to coalesce. However, with appropriate tuning of the thresholds for detection and length of data collection, we clearly observed 2 distinct states of BBL, with FRET efficiencies corresponding to native and denatured states. The population of each state varied with GdmCl or urea during chemical denaturation transitions corresponding to conventional barrier-limited folding at 279 K and pH 7 and pH 5.8. The folding of BBL is accordingly barrier limited. PMID- 19805288 TI - Clustering of alpha(5)beta(1) integrins determines adhesion strength whereas alpha(v)beta(3) and talin enable mechanotransduction. AB - A key molecular link between cells and the extracellular matrix is the binding between fibronectin and integrins alpha(5)beta(1) and alpha(v)beta(3). However, the roles of these different integrins in establishing adhesion remain unclear. We tested the adhesion strength of fibronectin-integrin-cytoskeleton linkages by applying physiological nanonewton forces to fibronectin-coated magnetic beads bound to cells. We report that the clustering of fibronectin domains within 40 nm led to integrin alpha(5)beta(1) recruitment, and increased the ability to sustain force by over six-fold. This force was supported by alpha(5)beta(1) integrin clusters. Importantly, we did not detect a role of either integrin alpha(v)beta(3) or talin 1 or 2 in maintaining adhesion strength. Instead, these molecules enabled the connection to the cytoskeleton and reinforcement in response to an applied force. Thus, high matrix forces are primarily supported by clustered alpha(5)beta(1) integrins, while less stable links to alpha(v)beta(3) integrins initiate mechanotransduction, resulting in reinforcement of integrin cytoskeleton linkages through talin-dependent bonds. PMID- 19805289 TI - Structure of the C-terminus of the mRNA export factor Dbp5 reveals the interaction surface for the ATPase activator Gle1. AB - The DExD/H-box RNA-dependent ATPase Dbp5 plays an essential role in the nuclear export of mRNA. Dbp5 localizes to the nuclear pore complex, where its ATPase activity is stimulated by Gle1 and its coactivator inositol hexakisphosphate. Here, we present the crystal structure of the C-terminal domain of Dbp5, refined to 1.8 A. The structure reveals a RecA-like fold that contains two defining characteristics not present in other structurally characterized DExD/H-box proteins: a C-terminal alpha-helix and a loop connecting beta5 and alpha4, both of which are composed of conserved and unique elements in the Dbp5 primary sequence. Using structure-guided mutagenesis, we have identified several charged surface residues that, when mutated, weaken the binding of Gle1 and inhibit the ability of Gle1 to stimulate Dbp5's ATPase activity. In vivo analysis of the same mutations reveals that those mutants displaying the weakest ATPase stimulation in vitro are also unable to support yeast growth. Analysis of the correlation between the in vitro and in vivo data indicates that a threshold level of Dbp5 ATPase activity is required for cellular mRNA export that is not met by the unstimulated enzyme, suggesting a possible mechanism by which Dbp5's activity can be modulated to regulate mRNA export. PMID- 19805290 TI - Real-time imaging of histone H4 hyperacetylation in living cells. AB - To visualize histone acetylation in living cells, we developed a genetically encoded fluorescent resonance energy transfer (FRET)-based indicator. Response of the indicator reflects changes in the acetylation state of both K5 and K8 in histone H4. Using this acetylation indicator, we were able to monitor the dynamic fluctuation of histone H4 acetylation levels during mitosis, as well as acetylation changes in response to structurally distinct histone deacetylase inhibitors. PMID- 19805291 TI - Abcb10 physically interacts with mitoferrin-1 (Slc25a37) to enhance its stability and function in the erythroid mitochondria. AB - Mitoferrin-1 (Mfrn1; Slc25a37), a member of the solute carrier family localized in the mitochondrial inner membrane, functions as an essential iron importer for the synthesis of mitochondrial heme and iron-sulfur clusters in erythroblasts. The biochemistry of Mfrn1-mediated iron transport into the mitochondria, however, is poorly understood. Here, we used the strategy of in vivo epitope-tagging affinity purification and mass spectrometry to investigate Mfrn1-mediated mitochondrial iron homeostasis. Abcb10, a mitochondrial inner membrane ATP binding cassette transporter highly induced during erythroid maturation in hematopoietic tissues, was found as one key protein that physically interacts with Mfrn1 during mouse erythroleukemia (MEL) cell differentiation. Mfrn1 was shown previously to have a longer protein half-life in differentiated MEL cells compared with undifferentiated cells. In this study, Abcb10 was found to enhance the stabilization of Mfrn1 protein in MEL cells and transfected heterologous COS7 cells. In undifferentiated MEL cells, cotransfected Abcb10 specifically interacts with Mfrn1 to enhance its protein stability and promote Mfrn1-dependent mitochondrial iron importation. The structural stabilization of the Mfrn1-Abcb10 complex demonstrates a previously uncharacterized function for Abcb10 in mitochondria. Furthermore, the binding domain of Mfrn1-Abcb10 interaction maps to the N terminus of Mfrn1. These results suggest the tight regulation of mitochondrial iron acquisition and heme synthesis in erythroblasts is mediated by both transcriptional and posttranslational mechanisms, whereby the high level of Mfrn1 is stabilized by oligomeric protein complexes. PMID- 19805292 TI - The interaction of nucleoside diphosphate kinase B with Gbetagamma dimers controls heterotrimeric G protein function. AB - Heterotrimeric G proteins in physiological and pathological processes have been extensively studied so far. However, little is known about mechanisms regulating the cellular content and compartmentalization of G proteins. Here, we show that the association of nucleoside diphosphate kinase B (NDPK B) with the G protein betagamma dimer (Gbetagamma) is required for G protein function in vivo. In zebrafish embryos, morpholino-mediated knockdown of zebrafish NDPK B, but not NDPK A, results in a severe decrease in cardiac contractility. The depletion of NDPK B is associated with a drastic reduction in Gbeta(1)gamma(2) dimer expression. Moreover, the protein levels of the adenylyl cyclase (AC)-regulating Galpha(s) and Galpha(i) subunits as well as the caveolae scaffold proteins caveolin-1 and -3 are strongly reduced. In addition, the knockdown of the zebrafish Gbeta(1) orthologs, Gbeta(1) and Gbeta(1like), causes a cardiac phenotype very similar to that of NDPK B morphants. The loss of Gbeta(1)/Gbeta(1like) is associated with a down-regulation in caveolins, AC regulating Galpha-subunits, and most important, NDPK B. A comparison of embryonic fibroblasts from wild-type and NDPK A/B knockout mice demonstrate a similar reduction of G protein, caveolin-1 and basal cAMP content in mammalian cells that can be rescued by re-expression of human NDPK B. Thus, our results suggest a role for the interaction of NDPK B with Gbetagamma dimers and caveolins in regulating membranous G protein content and maintaining normal G protein function in vivo. PMID- 19805293 TI - CBP and p300 are cytoplasmic E4 polyubiquitin ligases for p53. AB - p300 and CREB-binding protein (CBP) act as multifunctional regulators of p53 via acetylase and polyubiquitin ligase (E4) activities. Prior work in vitro has shown that the N-terminal 595 aa of p300 encode both generic ubiquitin ligase (E3) and p53-directed E4 functions. Analysis of p300 or CBP-deficient cells revealed that both coactivators were required for endogenous p53 polyubiquitination and the normally rapid turnover of p53 in unstressed cells. Unexpectedly, p300/CBP ubiquitin ligase activities were absent in nuclear extracts and exclusively cytoplasmic. Consistent with the cytoplasmic localization of its E3/E4 activity, CBP deficiency specifically stabilized cytoplasmic, but not nuclear p53. The N terminal 616 aa of CBP, which includes the conserved Zn(2+)-binding C/H1-TAZ1 domain, was the minimal domain sufficient to destabilize p53 in vivo, and it included within an intrinsic E3 autoubiquitination activity and, in a two-step E4 assay, exhibited robust E4 activity for p53. Cytoplasmic compartmentalization of p300/CBP's ubiquitination function reconciles seemingly opposed functions and explains how a futile cycle is avoided-cytoplasmic p300/CBP E4 activities ubiquitinate and destabilize p53, while physically separate nuclear p300/CBP activities, such as p53 acetylation, activate p53. PMID- 19805294 TI - Highly tumorigenic lung cancer CD133+ cells display stem-like features and are spared by cisplatin treatment. AB - The identification of lung tumor-initiating cells and associated markers may be useful for optimization of therapeutic approaches and for predictive and prognostic information in lung cancer patients. CD133, a surface glycoprotein linked to organ-specific stem cells, was described as a marker of cancer initiating cells in different tumor types. Here, we report that a CD133+, epithelial-specific antigen-positive (CD133+ESA+) population is increased in primary nonsmall cell lung cancer (NSCLC) compared with normal lung tissue and has higher tumorigenic potential in SCID mice and expression of genes involved in stemness, adhesion, motility, and drug efflux than the CD133(-) counterpart. Cisplatin treatment of lung cancer cells in vitro resulted in enrichment of CD133+ fraction both after acute cytotoxic exposure and in cells with stable cisplatin-resistant phenotype. Subpopulations of CD133+ABCG2+ and CD133+CXCR4+ cells were spared by in vivo cisplatin treatment of lung tumor xenografts established from primary tumors. A tendency toward shorter progression-free survival was observed in CD133+ NSCLC patients treated with platinum-containing regimens. Our results indicate that chemoresistant populations with highly tumorigenic and stem-like features are present in lung tumors. The molecular features of these cells may provide the rationale for more specific therapeutic targeting and the definition of predictive factors in clinical management of this lethal disease. PMID- 19805296 TI - Adaptive shell color plasticity during the early ontogeny of an intertidal keystone snail. AB - We report a mechanism of crypsis present during the vulnerable early post metamorphic ontogeny (95%) of specimens bearing patterns of shell coloration (dark or light colored) that matched the background coloration provided by patches of Concholepas' most abundant prey (mussels or barnacles respectively). The variation in shell color was positively associated with the color of the most common prey (r = 0.99). In laboratory experiments, shell coloration of C. concholepas depended on the prey substrate used to induce metamorphosis and for the post-metamorphic rearing. The snail shell color matched the color of the prey offered during rearing. Laboratory manipulation experiments, switching the prey during rearing, showed a corresponding change in snail shell color along the outermost shell edge. As individuals grew and became increasingly indistinguishable from the surrounding background, cryptic individuals had higher survival (71%) than the non cryptic ones (4%) when they were reared in the presence of the predatory crab Acanthocyclus hassleri. These results suggest that the evolution of shell color plasticity during the early ontogeny of C. concholepas, depends on the color of the more abundant of the consumed prey available in the natural habitat where settlement has taken place; this in turn has important consequences for their fitness and survivorship in the presence of visual predators. PMID- 19805295 TI - beta-Catenin promotes respiratory progenitor identity in mouse foregut. AB - The mammalian respiratory system, consisting of both trachea and lung, initiates from the foregut endoderm. The molecular program that instructs endodermal cells to adopt the respiratory fate is not fully understood. Here we show that conditional inactivation of beta-Catenin (also termed Ctnnb1) in foregut endoderm leads to absence of both the trachea and lung due to a failure in maintaining the respiratory fate. In converse, conditional expression of an activated form of beta-Catenin leads to expansion of Nkx2.1, an early marker for the trachea and lung, into adjacent endoderm including the stomach epithelium. Analyses of these mutants show that the loss or gain of trachea/lung progenitor identity is accompanied by an expansion or contraction of esophagus/stomach progenitor identity, respectively. Our findings reveal an early role for beta-Catenin in the establishment of respiratory progenitors in mouse foregut endoderm. PMID- 19805297 TI - The phytohormone precursor OPDA is isomerized in the insect gut by a single, specific glutathione transferase. AB - Oxylipins play important roles in stress signaling in plants. The compound 12 oxophytodienoic acid (cis-OPDA) is an early biosynthetic precursor of jasmonic acid (JA), the key phytohormone orchestrating the plant anti-herbivore defense. When consumed by feeding Lepidopteran larvae, plant-derived cis-OPDA suffers rapid isomerization to iso-OPDA in the midgut and is excreted in the frass. Unlike OPDA epimerization (yielding trans-OPDA), the formation of iso-OPDA is enzyme-dependent, and is catalyzed by an inducible glutathione transferase (GSTs) from the larval gut. Purified GST fractions from the gut of Egyptian cotton leafworm (Spodoptera littoralis) and cotton bollworm (Helicoverpa armigera) both exhibited strong OPDA isomerization activity, most likely via transient formation of a glutathione-OPDA conjugate. Out of 16 cytosolic GST proteins cloned from the gut of cotton bollworm larvae and expressed in E. coli, only one catalyzed the OPDA isomerization. The biological function of the double bond shift might be seen in an inactivation of cis-OPDA, similar to the inactivation of prostaglandin A1 to prostaglandin B1 in mammalian tissue. The enzymatic isomerization is particularly widespread among generalist herbivores that have to cope with various amounts of cis-OPDA in their spectrum of host plants. PMID- 19805298 TI - A genome-wide view of Caenorhabditis elegans base-substitution mutation processes. AB - Knowledge of mutation processes is central to understanding virtually all evolutionary phenomena and the underlying nature of genetic disorders and cancers. However, the limitations of standard molecular mutation detection methods have historically precluded a genome-wide understanding of mutation rates and spectra in the nuclear genomes of multicellular organisms. We applied two high-throughput DNA sequencing technologies to identify and characterize hundreds of spontaneously arising base-substitution mutations in 10 Caenorhabditis elegans mutation-accumulation (MA)-line nuclear genomes. C. elegans mutation rate estimates were similar to previous calculations based on smaller numbers of mutations. Mutations were distributed uniformly within and among chromosomes and were not associated with recombination rate variation in the MA lines, suggesting that intragenomic variation in genetic hitchhiking and/or background selection are primarily responsible for the chromosomal distribution patterns of polymorphic nucleotides in C. elegans natural populations. A strong mutational bias from G/C to A/T nucleotides was detected in the MA lines, implicating oxidative DNA damage as a major endogenous mutagenic force in C. elegans. The observed mutational bias also suggests that the C. elegans nuclear genome cannot be at equilibrium because of mutation alone. Transversions dominate the spectrum of spontaneous mutations observed here, whereas transitions dominate patterns of allegedly neutral polymorphism in natural populations of C. elegans and many other animal species; this observation challenges the assumption that natural patterns of molecular variation in noncoding regions of the nuclear genome accurately reflect underlying mutation processes. PMID- 19805299 TI - Post-Pleistocene radiation of the pea aphid complex revealed by rapidly evolving endosymbionts. AB - Adaptation to different resources has the potential to cause rapid species diversification, but few studies have been able to quantify the time scale of recent adaptive radiations. The pea aphid, Acyrthosiphon pisum, a model of speciation for host-specialized parasites, consists of several biotypes (races or species) living on distinct legume hosts. To document this radiation, we used rapidly evolving sequences from Buchnera, the maternally transmitted bacterial endosymbiont of aphids. Analyses of Buchnera pseudogene sequences revealed that 11 host-associated biotypes sort mostly into distinct matrilines despite low sequence divergence. A calibration based on divergence times of 7 sequenced genomes of Buchnera allowed us to date the last maternal ancestor of these biotypes between 8,000 and 16,000 years, with a burst of diversification at an estimated 3,600-9,500 years. The recency of this diversification, which is supported by microsatellite data, implies that the pea aphid complex ranks among the most rapid adaptive radiations yet documented. This diversification coincides with post-Pleistocene warming and with the domestication and anthropogenic range expansion of several of the legume hosts of pea aphids. Thus, we hypothesize that the new availability or abundance of resources triggered a cascade of divergence events in this newly formed complex. PMID- 19805300 TI - A polycomb repressive complex 2 gene regulates apogamy and gives evolutionary insights into early land plant evolution. AB - Land plants have distinct developmental programs in haploid (gametophyte) and diploid (sporophyte) generations. Although usually the two programs strictly alternate at fertilization and meiosis, one program can be induced during the other program. In a process called apogamy, cells of the gametophyte other than the egg cell initiate sporophyte development. Here, we report for the moss Physcomitrella patens that apogamy resulted from deletion of the gene orthologous to the Arabidopsis thaliana CURLY LEAF (PpCLF), which encodes a component of polycomb repressive complex 2 (PRC2). In the deletion lines, a gametophytic vegetative cell frequently gave rise to a sporophyte-like body. This body grew indeterminately from an apical cell with the character of a sporophytic pluripotent stem cell but did not form a sporangium. Furthermore, with continued culture, the sporophyte-like body branched. Sporophyte branching is almost unknown among extant bryophytes. When PpCLF was expressed in the deletion lines once the sporophyte-like bodies had formed, pluripotent stem cell activity was arrested and a sporangium-like organ formed. Supported by the observed pattern of PpCLF expression, these results demonstrate that, in the gametophyte, PpCLF represses initiation of a sporophytic pluripotent stem cell and, in the sporophyte, represses that stem cell activity and induces reproductive organ development. In land plants, branching, along with indeterminate apical growth and delayed initiation of spore-bearing reproductive organs, were conspicuous innovations for the evolution of a dominant sporophyte plant body. Our study provides insights into the role of PRC2 gene regulation for sustaining evolutionary innovation in land plants. PMID- 19805301 TI - Elephant shark (Callorhinchus milii) provides insights into the evolution of Hox gene clusters in gnathostomes. AB - We have sequenced and analyzed Hox gene clusters from elephant shark, a holocephalian cartilaginous fish. Elephant shark possesses 4 Hox clusters with 45 Hox genes that include orthologs for a higher number of ancient gnathostome Hox genes than the 4 clusters in tetrapods and the supernumerary clusters in teleost fishes. Phylogenetic analysis of elephant shark Hox genes from 7 paralogous groups that contain all of the 4 members indicated an ((AB)(CD)) topology for the order of Hox cluster duplication, providing support for the 2R hypothesis (i.e., 2 rounds of whole-genome duplication during the early evolution of vertebrates). Comparisons of noncoding sequences of the elephant shark and human Hox clusters have identified a large number of conserved noncoding elements (CNEs), which represent putative cis-regulatory elements that may be involved in the regulation of Hox genes. Interestingly, in fugu more than 50% of these ancient CNEs have diverged beyond recognition in the duplicated (HoxA, HoxB, and HoxD) as well as the singleton (HoxC) Hox clusters. Furthermore, the b-paralogs of the duplicated fugu Hox clusters are virtually devoid of unique ancient CNEs. In contrast to fugu Hox clusters, elephant shark and human Hox clusters have lost fewer ancient CNEs. If these ancient CNEs are indeed enhancers directing tissue-specific expression of Hox genes, divergence of their sequences in vertebrate lineages might have led to altered expression patterns and presumably the functions of their associated Hox genes. PMID- 19805302 TI - Eukaryote-to-eukaryote gene transfer events revealed by the genome sequence of the wine yeast Saccharomyces cerevisiae EC1118. AB - Saccharomyces cerevisiae has been used for millennia in winemaking, but little is known about the selective forces acting on the wine yeast genome. We sequenced the complete genome of the diploid commercial wine yeast EC1118, resulting in an assembly of 31 scaffolds covering 97% of the S288c reference genome. The wine yeast differed strikingly from the other S. cerevisiae isolates in possessing 3 unique large regions, 2 of which were subtelomeric, the other being inserted within an EC1118 chromosome. These regions encompass 34 genes involved in key wine fermentation functions. Phylogeny and synteny analyses showed that 1 of these regions originated from a species closely related to the Saccharomyces genus, whereas the 2 other regions were of non-Saccharomyces origin. We identified Zygosaccharomyces bailii, a major contaminant of wine fermentations, as the donor species for 1 of these 2 regions. Although natural hybridization between Saccharomyces strains has been described, this report provides evidence that gene transfer may occur between Saccharomyces and non-Saccharomyces species. We show that the regions identified are frequent and differentially distributed among S. cerevisiae clades, being found almost exclusively in wine strains, suggesting acquisition through recent transfer events. Overall, these data show that the wine yeast genome is subject to constant remodeling through the contribution of exogenous genes. Our results suggest that these processes are favored by ecologic proximity and are involved in the molecular adaptation of wine yeasts to conditions of high sugar, low nitrogen, and high ethanol concentrations. PMID- 19805303 TI - Validation-based insertional mutagenesis identifies lysine demethylase FBXL11 as a negative regulator of NFkappaB. AB - We describe a highly efficient use of lentiviral validation-based insertional mutagenesis (VBIM) to generate large populations of mammalian cells in which a strong promoter is inserted into many different genomic loci, causing greatly increased expression of downstream sequences. Many different selections or screens can follow, to isolate dominant mutant clones with a desired phenotypic change. The inserted promoter can be excised or silenced at will, to prove that the insertion caused the mutation. Cloning DNA flanking the insertion site identifies the locus precisely. VBIM virus particles are pseudotyped with VSV G protein, allowing efficient infection of most mammalian cell types, including non dividing cells, and features are included that give high yields of stable virus stocks. In several different selections, useful mutants have been obtained at frequencies of approximately 10(-6) or higher. We used the VBIM technique to isolate mutant human cells in which the F-box leucine-rich protein 11 (FBXL11), a histone H3K36 demethylase, is shown to be a negative regulator of NFkappaB. High levels of FBXL11 block the ability of NFkappaB to bind to DNA or activate gene expression, and siRNA-mediated reduction of FBXL11 expression has the opposite effects. The H212A mutation of FBXL11 abolishes both its histone H3K36 demethylase activity and its ability to inhibit NFkappaB. Thus, we have used a powerful tool for mutagenesis of mammalian cells to reveal an aspect of the complex regulation of NFkappaB-dependent signaling. PMID- 19805304 TI - Gasp, a Grb2-associating protein, is critical for positive selection of thymocytes. AB - T cells develop in the thymus through positive and negative selection, which are responsible for shaping the T cell receptor (TCR) repertoire. To elucidate the molecular mechanisms involved in selection remains an area of intense interest. Here, we identified and characterized a gene product Gasp (Grb2-associating protein, also called Themis) that is critically required for positive selection. Gasp is a cytosolic protein with no known functional motifs that is expressed only in T cells, especially immature CD4/CD8 double positive (DP) thymocytes. In the absence of Gasp, differentiation of both CD4 and CD8 single positive cells in the thymus was severely inhibited, whereas all other TCR-induced events such as beta-selection, negative selection, peripheral activation, and homeostatic proliferation were unaffected. We found that Gasp constitutively associates with Grb2 via its N-terminal Src homology 3 domain, suggesting that Gasp acts as a thymocyte-specific adaptor for Grb2 or regulates Ras signaling in DP thymocytes. Collectively, we have described a gene called Gasp that is critical for positive selection. PMID- 19805305 TI - XMRV is present in malignant prostatic epithelium and is associated with prostate cancer, especially high-grade tumors. AB - Xenotropic murine leukemia virus-related virus (XMRV) was recently discovered in human prostate cancers and is the first gammaretrovirus known to infect humans. While gammaretroviruses have well-characterized oncogenic effects in animals, they have not been shown to cause human cancers. We provide experimental evidence that XMRV is indeed a gammaretrovirus with protein composition and particle ultrastructure highly similar to Moloney murine leukemia virus (MoMLV), another gammaretrovirus. We analyzed 334 consecutive prostate resection specimens, using a quantitative PCR assay and immunohistochemistry (IHC) with an anti-XMRV specific antiserum. We found XMRV DNA in 6% and XMRV protein expression in 23% of prostate cancers. XMRV proteins were expressed primarily in malignant epithelial cells, suggesting that retroviral infection may be directly linked to tumorigenesis. XMRV infection was associated with prostate cancer, especially higher-grade cancers. We found XMRV infection to be independent of a common polymorphism in the RNASEL gene, unlike results previously reported. This finding increases the population at risk for XMRV infection from only those homozygous for the RNASEL variant to all individuals. Our observations provide evidence for an association of XMRV with malignant cells and with more aggressive tumors. PMID- 19805307 TI - Positron emission tomography imaging of drug-induced tumor apoptosis with a caspase-3/7 specific [18F]-labeled isatin sulfonamide. AB - Of the molecular biochemical alterations that occur during apoptosis, activation of caspases, notably caspase-3, is probably the most attractive for developing specific in vivo molecular imaging probes. We recently designed a library of isatin-5 sulfonamides and selected [18F]ICMT-11 for further evaluation on the basis of subnanomolar affinity for activated capsase-3, high metabolic stability, and facile radiolabeling. In this present study, we have demonstrated that [18F]ICMT-11 binds to a range of drug-induced apoptotic cancer cells in vitro and to 38C13 murine lymphoma xenografts in vivo by up to 2-fold at 24 h posttreatment compared to vehicle treatment. We further demonstrated that the increased signal intensity in tumors after drug treatment, detected by whole body in vivo microPET imaging, was associated with increased apoptosis. In summary, we have characterized [18F]ICMT-11 as a caspase-3/7 specific PET imaging radiotracer for the assessment of tumor apoptosis that could find utility in anticancer drug development and the monitoring of early responses to therapy. PMID- 19805306 TI - aPKClambda/iota promotes growth of prostate cancer cells in an autocrine manner through transcriptional activation of interleukin-6. AB - Understanding the mechanism by which hormone refractory prostate cancer (HRPC) develops remains a major issue. Alterations in HRPC include androgen receptor (AR) changes. In addition, the AR is activated by cytokines such as interleukin-6 (IL-6). Atypical protein kinase C (aPKClambda/iota) has been implicated in the progression of several cancers. Herein, we provide evidence that aPKClambda/iota expression correlates with prostate cancer recurrence. Experiments in vitro and in vivo revealed aPKClambda/iota to be involved in prostate cancer cell growth through secretion of IL-6. Further, aPKClambda/iota activates transcription of the IL-6 gene through NFkappaB and AP-1. We conclude that aPKClambda/iota promotes the growth of hormone independent prostate cancer cells by stimulating IL-6 production in an autocrine manner. Our findings not only explain the link between aPKClambda/iota and IL-6, implicated in the progression a variety of cancers, but also establish a molecular change involved in the development of HRPC. Further, aPKClambda/iota expression might be a biomarker for prostate cancer progression. PMID- 19805308 TI - Elucidation of the mechanism of mitochondrial iron loading in Friedreich's ataxia by analysis of a mouse mutant. AB - We used the muscle creatine kinase (MCK) conditional frataxin knockout mouse to elucidate how frataxin deficiency alters iron metabolism. This is of significance because frataxin deficiency leads to Friedreich's ataxia, a disease marked by neurologic and cardiologic degeneration. Using cardiac tissues, we demonstrate that frataxin deficiency leads to down-regulation of key molecules involved in 3 mitochondrial utilization pathways: iron-sulfur cluster (ISC) synthesis (iron sulfur cluster scaffold protein1/2 and the cysteine desulferase Nfs1), mitochondrial iron storage (mitochondrial ferritin), and heme synthesis (5 aminolevulinate dehydratase, coproporphyrinogen oxidase, hydroxymethylbilane synthase, uroporphyrinogen III synthase, and ferrochelatase). This marked decrease in mitochondrial iron utilization and resultant reduced release of heme and ISC from the mitochondrion could contribute to the excessive mitochondrial iron observed. This effect is compounded by increased iron availability for mitochondrial uptake through (i) transferrin receptor1 up-regulation, increasing iron uptake from transferrin; (ii) decreased ferroportin1 expression, limiting iron export; (iii) increased expression of the heme catabolism enzyme heme oxygenase1 and down-regulation of ferritin-H and -L, both likely leading to increased "free iron" for mitochondrial uptake; and (iv) increased expression of the mammalian exocyst protein Sec15l1 and the mitochondrial iron importer mitoferrin-2 (Mfrn2), which facilitate cellular iron uptake and mitochondrial iron influx, respectively. Our results enable the construction of a model explaining the cytosolic iron deficiency and mitochondrial iron loading in the absence of frataxin, which is important for understanding the pathogenesis of Friedreich's ataxia. PMID- 19805309 TI - Genetic heterogeneity of Myc-induced mammary tumors reflecting diverse phenotypes including metastatic potential. AB - Human cancers result from a complex series of genetic alterations, resulting in heterogeneous disease states. Dissecting this heterogeneity is critical for understanding underlying mechanisms and providing opportunities for therapeutics matching the complexity. Mouse models of cancer have generally been used to reduce this complexity and focus on the role of single genes. Nevertheless, our analysis of tumors arising in the MMTV-Myc model of mammary carcinogenesis reveals substantial heterogeneity, seen in both histological and expression phenotypes. One contribution to this heterogeneity is the substantial frequency of activating Ras mutations. Additionally, we show that these Myc-induced mammary tumors exhibit even greater heterogeneity, revealed by distinct histological subtypes as well as distinct patterns of gene expression, than many other mouse models of tumorigenesis. Two of the major histological subtypes are characterized by differential patterns of cellular signaling pathways, including beta-catenin and Stat3 activities. We also demonstrate that one of the MMTV-Myc mammary tumor subgroups exhibits metastatic capacity and that the signature derived from the subgroup can predict metastatic potential of human breast cancer. Together, these data reveal that a combination of histological and genomic analyses can uncover substantial heterogeneity in mammary tumor formation and therefore highlight aspects of tumor phenotype not evident in the population as a whole. PMID- 19805311 TI - Spatiotemporal patterns and transcription kinetics of induced RNA in single bacterial cells. AB - Bacteria have a complex internal organization with specific localization of many proteins and DNA, which dynamically move during the cell cycle and in response to changing environmental stimuli. Much less is known, however, about the localization and movements of RNA molecules. By modifying our previous RNA labeling system, we monitor the expression and localization of a model RNA transcript in live Escherichia coli cells. Our results reveal that the target RNA is not evenly distributed within the cell and localizes laterally along the long cell axis, in a pattern suggesting the existence of ordered helical RNA structures reminiscent of known bacterial cytoskeletal cellular elements. PMID- 19805312 TI - Subcellular localization of a bacterial regulatory RNA. AB - Eukaryotes and bacteria regulate the activity of some proteins by localizing them to discrete subcellular structures, and eukaryotes localize some RNAs for the same purpose. To explore whether bacteria also spatially regulate RNAs, the localization of tmRNA was determined using fluorescence in situ hybridization. tmRNA is a small regulatory RNA that is ubiquitous in bacteria and that interacts with translating ribosomes in a reaction known as trans-translation. In Caulobacter crescentus, tmRNA was localized in a cell-cycle-dependent manner. In G(1)-phase cells, tmRNA was found in regularly spaced foci indicative of a helix like structure. After initiation of DNA replication, most of the tmRNA was degraded, and the remaining molecules were spread throughout the cytoplasm. Immunofluorescence assays showed that SmpB, a protein that binds tightly to tmRNA, was colocalized with tmRNA in the helix-like pattern. RNase R, the nuclease that degrades tmRNA, was localized in a helix-like pattern that was separate from the SmpB-tmRNA complex. These results suggest a model in which tmRNA-SmpB is localized to sequester tmRNA from RNase R, and localization might also regulate tmRNA-SmpB interactions with ribosomes. PMID- 19805313 TI - Kinetic control of TolC recruitment by multidrug efflux complexes. AB - In Gram-negative pathogens, multidrug efflux pumps that provide clinically significant levels of antibiotic resistance function as three-component complexes. They are composed of the inner membrane transporters belonging to one of three superfamilies of proteins, RND, ABC, or MF; periplasmic proteins belonging to the membrane fusion protein (MFP) family; and outer membrane channels exemplified by the Escherichia coli TolC. The three-component complexes span the entire two-membrane envelope of Gram-negative bacteria and expel toxic molecules from the cytoplasmic membrane to the medium. The architecture of these complexes is expected to vary significantly because of the structural diversity of the inner membrane transporters. How the three-component pumps are assembled, their architecture, and their dynamics remain unclear. In this study, we reconstituted interactions and compared binding kinetics of the E. coli TolC with AcrA, MacA, and EmrA, the periplasmic MFPs that function in multidrug efflux with transporters from the RND, ABC, and MF superfamilies, respectively. By using surface plasmon resonance, we demonstrate that TolC interactions with MFPs are highly dynamic and sensitive to pH. The affinity of TolC to MFPs decreases in the order MacA > EmrA > AcrA. We further show that MFPs are prone to oligomerization, but differ dramatically from each other in oligomerization kinetics and stability of oligomers. The propensity of MFPs to oligomerize correlates with the stability of MFP-TolC complexes and structural features of inner membrane transporters. We propose that recruitment of TolC by various MFPs is determined not only by kinetics of MFP-TolC interactions but also by oligomerization kinetics of MFPs and pH. PMID- 19805314 TI - Tracking insertion mutants within libraries by deep sequencing and a genome-wide screen for Haemophilus genes required in the lung. AB - Rapid genome-wide identification of genes required for infection would expedite studies of bacterial pathogens. We developed genome-scale "negative selection" technology that combines high-density transposon mutagenesis and massively parallel sequencing of transposon/chromosome junctions in a mutant library to identify mutants lost from the library after exposure to a selective condition of interest. This approach was applied to comprehensively identify Haemophilus influenzae genes required to delay bacterial clearance in a murine pulmonary model. Mutations in 136 genes resulted in defects in vivo, and quantitative estimates of fitness generated by this technique were in agreement with independent validation experiments using individual mutant strains. Genes required in the lung included those with characterized functions in other models of H. influenzae pathogenesis and genes not previously implicated in infection. Genes implicated in vivo have reported or potential roles in survival during nutrient limitation, oxidative stress, and exposure to antimicrobial membrane perturbations, suggesting that these conditions are encountered by H. influenzae during pulmonary infection. The results demonstrate an efficient means to identify genes required for bacterial survival in experimental models of pathogenesis, and this approach should function similarly well in selections conducted in vitro and in vivo with any organism amenable to insertional mutagenesis. PMID- 19805316 TI - Diminished dosage of 22q11 genes disrupts neurogenesis and cortical development in a mouse model of 22q11 deletion/DiGeorge syndrome. AB - The 22q11 deletion (or DiGeorge) syndrome (22q11DS), the result of a 1.5- to 3 megabase hemizygous deletion on human chromosome 22, results in dramatically increased susceptibility for "diseases of cortical connectivity" thought to arise during development, including schizophrenia and autism. We show that diminished dosage of the genes deleted in the 1.5-megabase 22q11 minimal critical deleted region in a mouse model of 22q11DS specifically compromises neurogenesis and subsequent differentiation in the cerebral cortex. Proliferation of basal, but not apical, progenitors is disrupted, and subsequently, the frequency of layer 2/3, but not layer 5/6, projection neurons is altered. This change is paralleled by aberrant distribution of parvalbumin-labeled interneurons in upper and lower cortical layers. Deletion of Tbx1 or Prodh (22q11 genes independently associated with 22q11DS phenotypes) does not similarly disrupt basal progenitors. However, expression analysis implicates additional 22q11 genes that are selectively expressed in cortical precursors. Thus, diminished 22q11 gene dosage disrupts cortical neurogenesis and interneuron migration. Such developmental disruption may alter cortical circuitry and establish vulnerability for developmental disorders, including schizophrenia and autism. PMID- 19805315 TI - Community proteogenomics reveals insights into the physiology of phyllosphere bacteria. AB - Aerial plant surfaces represent the largest biological interface on Earth and provide essential services as sites of carbon dioxide fixation, molecular oxygen release, and primary biomass production. Rather than existing as axenic organisms, plants are colonized by microorganisms that affect both their health and growth. To gain insight into the physiology of phyllosphere bacteria under in situ conditions, we performed a culture-independent analysis of the microbiota associated with leaves of soybean, clover, and Arabidopsis thaliana plants using a metaproteogenomic approach. We found a high consistency of the communities on the 3 different plant species, both with respect to the predominant community members (including the alphaproteobacterial genera Sphingomonas and Methylo bacterium) and with respect to their proteomes. Observed known proteins of Methylobacterium were to a large extent related to the ability of these bacteria to use methanol as a source of carbon and energy. A remarkably high expression of various TonB-dependent receptors was observed for Sphingomonas. Because these outer membrane proteins are involved in transport processes of various carbohydrates, a particularly large substrate utilization pattern for Sphingomonads can be assumed to occur in the phyllosphere. These adaptations at the genus level can be expected to contribute to the success and coexistence of these 2 taxa on plant leaves. We anticipate that our results will form the basis for the identification of unique traits of phyllosphere bacteria, and for uncovering previously unrecorded mechanisms of bacteria-plant and bacteria bacteria relationships. PMID- 19805317 TI - S-nitrosylation of stargazin regulates surface expression of AMPA-glutamate neurotransmitter receptors. AB - Synaptic plasticity is mediated by changes in the surface expression of AMPA receptors (AMPARs). Stargazin and related transmembrane AMPAR regulatory proteins have emerged as the principal regulators of AMPAR surface expression. Here, we show in heterologous cells and primary neurons that stargazin is physiologically S-nitrosylated, resulting in increased surface expression. S-nitrosylation of stargazin increases binding to the AMPAR subunit GluR1, causing increased surface expression of the AMPAR. NMDAR stimulation, well known to activate neuronal nitric oxide synthase, increases both nitrosylation of stargazin and its binding to AMPAR. Thus, S-nitrosylation of stargazin is a physiologic regulator of AMPAR surface expression. PMID- 19805318 TI - Spiking in primary somatosensory cortex during natural whisking in awake head restrained rats is cell-type specific. AB - Sensation involves active movement of sensory organs, but it remains unknown how position or movement of sensory organs is encoded in cortex. In the rat whisker system, each whisker is represented by an individual cortical (barrel) column. Here, we quantified in awake, head-fixed rats the impact of natural whisker movements on action potential frequencies of single (identified) neurons located in different layers of somatosensory (barrel) cortex. In all layers, we found only weak correlations between spiking and whisker position or velocity. Conversely, whisking significantly increased spiking rate in a subset of neurons located preferentially in layer 5A. This finding suggests that whisker movement could be encoded by population responses of neurons within all layers and by single slender-tufted pyramids in layer 5A. PMID- 19805319 TI - Metalloproteinase MT5-MMP is an essential modulator of neuro-immune interactions in thermal pain stimulation. AB - Peripheral interactions between nociceptive fibers and mast cells contribute to inflammatory pain, but little is known about mechanisms mediating neuro-immune communication. Here we show that metalloproteinase MT5-MMP (MMP-24) is an essential mediator of peripheral thermal nociception and inflammatory hyperalgesia. We report that MT5-MMP is expressed by CGRP-containing peptidergic nociceptors in dorsal root ganglia and that Mmp24-deficient mice display enhanced sensitivity to noxious thermal stimuli under basal conditions. Consistently, mutant peptidergic sensory neurons hyperinnervate the skin, a phenotype that correlates with changes in the regulated cleavage of the cell-cell adhesion molecule N-cadherin. In contrast to basal nociception, Mmp24(-/-) mice do not develop thermal hyperalgesia during inflammation, a phenotype that appears associated with alterations in N-cadherin-mediated cell-cell interactions between mast cells and sensory fibers. Collectively, our findings demonstrate an essential role of MT5-MMP in the development of dermal neuro-immune synapses and suggest that this metalloproteinase may be a target for pain control. PMID- 19805310 TI - Opportunistic pathogens enriched in showerhead biofilms. AB - The environments we humans encounter daily are sources of exposure to diverse microbial communities, some of potential concern to human health. In this study, we used culture-independent technology to investigate the microbial composition of biofilms inside showerheads as ecological assemblages in the human indoor environment. Showers are an important interface for human interaction with microbes through inhalation of aerosols, and showerhead waters have been implicated in disease. Although opportunistic pathogens commonly are cultured from shower facilities, there is little knowledge of either their prevalence or the nature of other microorganisms that may be delivered during shower usage. To determine the composition of showerhead biofilms and waters, we analyzed rRNA gene sequences from 45 showerhead sites around the United States. We find that variable and complex, but specific, microbial assemblages occur inside showerheads. Particularly striking was the finding that sequences representative of non-tuberculous mycobacteria (NTM) and other opportunistic human pathogens are enriched to high levels in many showerhead biofilms, >100-fold above background water contents. We conclude that showerheads may present a significant potential exposure to aerosolized microbes, including documented opportunistic pathogens. The health risk associated with showerhead microbiota needs investigation in persons with compromised immune or pulmonary systems. PMID- 19805320 TI - Analogous intermediate shape coding in vision and touch. AB - We recognize, understand, and interact with objects through both vision and touch. Conceivably, these two sensory systems encode object shape in similar ways, which could facilitate cross-modal communication. To test this idea, we studied single neurons in macaque monkey intermediate visual (area V4) and somatosensory (area SII) cortex, using matched shape stimuli. We found similar patterns of shape sensitivity characterized by tuning for curvature direction. These parallel tuning patterns imply analogous shape coding mechanisms in intermediate visual and somatosensory cortex. PMID- 19805321 TI - Cooperative synapse formation in the neocortex. AB - Neuron morphology plays an important role in defining synaptic connectivity. Clearly, only pairs of neurons with closely positioned axonal and dendritic branches can be synaptically coupled. For excitatory neurons in the cerebral cortex, such axo-dendritic oppositions, termed potential synapses, must be bridged by dendritic spines to form synaptic connections. To explore the rules by which synaptic connections are formed within the constraints imposed by neuron morphology, we compared the distributions of the numbers of actual and potential synapses between pre- and postsynaptic neurons forming different laminar projections in rat barrel cortex. Quantitative comparison explicitly ruled out the hypothesis that individual synapses between neurons are formed independently of each other. Instead, the data are consistent with a cooperative scheme of synapse formation where multiple-synaptic connections between neurons are stabilized while neurons that do not establish a critical number of synapses are not likely to remain synaptically coupled. PMID- 19805322 TI - Differential but convergent functions of Ca2+ binding to synaptotagmin-1 C2 domains mediate neurotransmitter release. AB - Neurotransmitter release is triggered by cooperative Ca2+-binding to the Ca2+ sensor protein synaptotagmin-1. Synaptotagmin-1 contains two C2 domains, referred to as the C2A and C2B domains, that bind Ca2+ with similar properties and affinities. However, Ca2+ binding to the C2A domain is not required for release, whereas Ca2+ binding to the C2B domain is essential for release. We now demonstrate that despite its expendability, Ca2+-binding to the C2A domain significantly contributes to the overall triggering of neurotransmitter release, and determines its Ca2+ cooperativity. Biochemically, Ca2+ induces more tight binding of the isolated C2A domain than of the isolated C2B domain to standard liposomes composed of phosphatidylcholine and phosphatidylserine. However, here we show that surprisingly, the opposite holds true when the double C2A/B-domain fragment of synaptotagmin-1 is used instead of isolated C2 domains, and when liposomes containing a physiological lipid composition are used. Under these conditions, Ca2+ binding to the C2B domain but not the C2A domain becomes the primary determinant of phospholipid binding. Thus, the unique requirement for Ca2+ binding to the C2B domain for synaptotagmin-1 in Ca2+-triggered neurotransmitter release may be accounted for, at least in part, by the unusual phospholipid-binding properties of its double C2A/B-domain fragment. PMID- 19805325 TI - Myosin II is required for interkinetic nuclear migration of neural progenitors. AB - Interkinetic nuclear migration (INM) is a hallmark of the polarized stem and progenitor cells in the ventricular zone (VZ) of the developing vertebrate CNS. INM is responsible for the pseudostratification of the VZ, a crucial aspect of brain evolution. The nuclear migration toward the apical centrosomes in G2 is thought to be a dynein-microtubule-based process. By contrast, the cytoskeletal machinery involved in the basally directed nuclear translocation away from the centrosome in G1 has been enigmatic. Studying the latter aspect of INM requires manipulation of the cytoskeleton without impairing mitosis and cytokinesis. To this end, we have established a culture system of mouse embryonic telencephalon that reproduces cortical development, and have applied it to explore a role of actomyosin in INM. Using the nonmuscle myosin II inhibitor blebbistatin at a low concentration at which neither cell cycle progression nor cytokinesis is impaired, we show that myosin II is required for the apical-to-basal (ap-->bl), ab-centrosomal INM. Myosin II activity is also necessary for the nuclear translocation during delamination of subventricular zone (SVZ) cells, a second, telencephalon-specific type of neural progenitor. Moreover, the inhibition of ab centrosomal INM changes the balance between VZ and SVZ progenitor cell fate. Our data suggest a unifying concept in which the actomyosin contraction underlying ab centrosomal INM sets the stage for the evolutionary increase in VZ pseudostratification and for SVZ progenitor delamination, a key process in cortical expansion. PMID- 19805324 TI - Dendritic trafficking of BDNF mRNA is mediated by translin and blocked by the G196A (Val66Met) mutation. AB - Alternatively spliced brain-derived neurotrophic factor (BDNF) transcripts are targeted to distinct cellular compartments in neurons but the mechanisms underlying this sorting are unknown. Although only some BDNF isoforms are targeted to dendrites, we have found that the coding region common to all BDNF transcripts contains a constitutively active dendritic targeting signal and that this signal is suppressed in transcripts containing exons 1 or 4, which are restricted to the cell soma and proximal dendrites. This dendritic targeting signal is mediated by translin, an RNA-binding protein implicated in RNA trafficking, and is disrupted by the G196A mutation associated with memory deficits and psychiatric disorders. Molecular modeling and mutational studies indicate that the G196A mutation blocks dendritic targeting of BDNF mRNA by disrupting its interaction with translin. These findings implicate abnormal dendritic trafficking of BDNF mRNA in the pathophysiology of neuropsychiatric disorders linked to the G196A mutation. PMID- 19805323 TI - Genetic perturbation of postsynaptic activity regulates synapse elimination in developing cerebellum. AB - In many parts of the vertebrate nervous system, synaptic connections are remodeled during early postnatal life. Neural activity plays an important role in regulating one such rearrangement, synapse elimination, in the developing neuromuscular system, but there is little direct evidence on roles of pre- or postsynaptic activity in regulating synapse elimination in the developing brain. To address this issue, we expressed a chloride channel-yellow fluorescent protein fusion in cerebellar Purkinje cells (PCs) of transgenic mice to decrease their excitability. We then assessed elimination of supernumerary climbing fiber inputs to PCs. Individual PCs are innervated by multiple climbing fibers at birth; all but one are eliminated during the first three postnatal weeks in wild-type mice, but multiple innervation persists for at least three months in the transgenic mice. The normal redistribution of climbing fiber synapses from PC somata to proximal dendrites was also blunted in transgenics. These results show that normal electrical activity of the postsynaptic cell is required for it to attain a mature innervation pattern. PMID- 19805326 TI - Intrinsic, nondeterministic circadian rhythm generation in identified mammalian neurons. AB - Circadian rhythms are modeled as reliable and self-sustained oscillations generated by single cells. The mammalian suprachiasmatic nucleus (SCN) keeps near 24-h time in vivo and in vitro, but the identity of the individual cellular pacemakers is unknown. We tested the hypothesis that circadian cycling is intrinsic to a unique class of SCN neurons by measuring firing rate or Period2 gene expression in single neurons. We found that fully isolated SCN neurons can sustain circadian cycling for at least 1 week. Plating SCN neurons at <100 cells/mm(2) eliminated synaptic inputs and revealed circadian neurons that contained arginine vasopressin (AVP) or vasoactive intestinal polypeptide (VIP) or neither. Surprisingly, arrhythmic neurons (nearly 80% of recorded neurons) also expressed these neuropeptides. Furthermore, neurons were observed to lose or gain circadian rhythmicity in these dispersed cell cultures, both spontaneously and in response to forskolin stimulation. In SCN explants treated with tetrodotoxin to block spike-dependent signaling, neurons gained or lost circadian cycling over many days. The rate of PERIOD2 protein accumulation on the previous cycle reliably predicted the spontaneous onset of arrhythmicity. We conclude that individual SCN neurons can generate circadian oscillations; however, there is no evidence for a specialized or anatomically localized class of cell-autonomous pacemakers. Instead, these results indicate that AVP, VIP, and other SCN neurons are intrinsic but unstable circadian oscillators that rely on network interactions to stabilize their otherwise noisy cycling. PMID- 19805327 TI - Predictable irregularities in retinal receptive fields. AB - Understanding how the nervous system achieves reliable performance using unreliable components is important for many disciplines of science and engineering, in part because it can suggest ways to lower the energetic cost of computing. In vision, retinal ganglion cells partition visual space into approximately circular regions termed receptive fields (RFs). Average RF shapes are such that they would provide maximal spatial resolution if they were centered on a perfect lattice. However, individual shapes have fine-scale irregularities. Here, we find that irregular RF shapes increase the spatial resolution in the presence of lattice irregularities from approximately 60% to approximately 92% of that possible for a perfect lattice. Optimization of RF boundaries around their fixed center positions reproduced experimental observations neuron-by-neuron. Our results suggest that lattice irregularities determine the shapes of retinal RFs and that similar algorithms can improve the performance of retinal prosthetics where substantial irregularities arise at their interface with neural tissue. PMID- 19805328 TI - Intrahippocampal injection of a lentiviral vector expressing Nrf2 improves spatial learning in a mouse model of Alzheimer's disease. AB - The amyloid hypothesis of Alzheimer's disease (AD) postulates that amyloid-beta (Abeta) deposition and neurotoxicity play a causative role in AD; oxidative injury is thought to be central in the pathogenesis. An endogenous defense system against oxidative stress is induced by binding of the transcription factor nuclear factor E2-related factor 2 (Nrf2) to the antioxidant response element (ARE) enhancer sequence. The Nrf2-ARE pathway is activated in response to reactive oxygen species to trigger the simultaneous expression of numerous protective enzymes and scavengers. To exploit the Nrf2-ARE pathway therapeutically, we delivered Nrf2 bilaterally into the hippocampus of 9-month old transgenic AD mice (APP/PS1 mice) using a lentiviral vector encoding human Nrf2. The data indicate that significant reductions in spatial learning deficits of aged APP/PS1 mice in a Morris Water Maze can be achieved by modulating levels of Nrf2 in the brain. Memory improvement in APP/PS1 mice after Nrf2 transduction shifts the balance between soluble and insoluble Abeta toward an insoluble Abeta pool without concomitant change in total brain Abeta burden. Nrf2 gene transfer is associated with a robust reduction in astrocytic but not microglial activation and induction of Nrf2 target gene heme oxygenase 1, indicating overall activation of the Nrf2-ARE pathway in hippocampal neurons 6 months after injection. Results warrant further exploration of the Nrf2-ARE pathway for treatment of AD and suggest that the Nrf2-ARE pathway may represent a potential therapeutic strategy to pursue in AD in humans, particularly in view of the multiple mechanisms by which Nrf2 can exert its protective effects. PMID- 19805329 TI - The putative cannabinoid receptor GPR55 affects osteoclast function in vitro and bone mass in vivo. AB - GPR55 is a G protein-coupled receptor recently shown to be activated by certain cannabinoids and by lysophosphatidylinositol (LPI). However, the physiological role of GPR55 remains unknown. Given the recent finding that the cannabinoid receptors CB(1) and CB(2) affect bone metabolism, we examined the role of GPR55 in bone biology. GPR55 was expressed in human and mouse osteoclasts and osteoblasts; expression was higher in human osteoclasts than in macrophage progenitors. Although the GPR55 agonists O-1602 and LPI inhibited mouse osteoclast formation in vitro, these ligands stimulated mouse and human osteoclast polarization and resorption in vitro and caused activation of Rho and ERK1/2. These stimulatory effects on osteoclast function were attenuated in osteoclasts generated from GPR55(-/-) macrophages and by the GPR55 antagonist cannabidiol (CBD). Furthermore, treatment of mice with this non-psychoactive constituent of cannabis significantly reduced bone resorption in vivo. Consistent with the ability of GPR55 to suppress osteoclast formation but stimulate osteoclast function, histomorphometric and microcomputed tomographic analysis of the long bones from male GPR55(-/-) mice revealed increased numbers of morphologically inactive osteoclasts but a significant increase in the volume and thickness of trabecular bone and the presence of unresorbed cartilage. These data reveal a role of GPR55 in bone physiology by regulating osteoclast number and function. In addition, this study also brings to light an effect of both the endogenous ligand, LPI, on osteoclasts and of the cannabis constituent, CBD, on osteoclasts and bone turnover in vivo. PMID- 19805330 TI - Molecular clock is involved in predictive circadian adjustment of renal function. AB - Renal excretion of water and major electrolytes exhibits a significant circadian rhythm. This functional periodicity is believed to result, at least in part, from circadian changes in secretion/reabsorption capacities of the distal nephron and collecting ducts. Here, we studied the molecular mechanisms underlying circadian rhythms in the distal nephron segments, i.e., distal convoluted tubule (DCT) and connecting tubule (CNT) and the cortical collecting duct (CCD). Temporal expression analysis performed on microdissected mouse DCT/CNT or CCD revealed a marked circadian rhythmicity in the expression of a large number of genes crucially involved in various homeostatic functions of the kidney. This analysis also revealed that both DCT/CNT and CCD possess an intrinsic circadian timing system characterized by robust oscillations in the expression of circadian core clock genes (clock, bma11, npas2, per, cry, nr1d1) and clock-controlled Par bZip transcriptional factors dbp, hlf, and tef. The clock knockout mice or mice devoid of dbp/hlf/tef (triple knockout) exhibit significant changes in renal expression of several key regulators of water or sodium balance (vasopressin V2 receptor, aquaporin-2, aquaporin-4, alphaENaC). Functionally, the loss of clock leads to a complex phenotype characterized by partial diabetes insipidus, dysregulation of sodium excretion rhythms, and a significant decrease in blood pressure. Collectively, this study uncovers a major role of molecular clock in renal function. PMID- 19805331 TI - Traditional models already explain adoption/abandonment pattern. PMID- 19805332 TI - Constraining future greenhouse gas emissions by a cumulative target. PMID- 19805333 TI - pH regulation in symbiotic anemones and corals: a delicate balancing act. PMID- 19805334 TI - Tales of retinogenesis told by human stem cells. PMID- 19805335 TI - Unraveling different chemical fingerprints between a champagne wine and its aerosols. AB - As champagne or sparkling wine is poured into a glass, the myriad of ascending bubbles collapse and radiate a multitude of tiny droplets above the free surface into the form of very characteristic and refreshing aerosols. Ultrahigh resolution MS was used as a nontargeted approach to discriminate hundreds of surface active compounds that are preferentially partitioning in champagne aerosols; thus, unraveling different chemical fingerprints between the champagne bulk and its aerosols. Based on accurate exact mass analysis and database search, tens of these compounds overconcentrating in champagne aerosols were unambiguously discriminated and assigned to compounds showing organoleptic interest or being aromas precursors. By drawing a parallel between the fizz of the ocean and the fizz in Champagne wines, our results closely link bursting bubbles and flavor release; thus, supporting the idea that rising and collapsing bubbles act as a continuous paternoster lift for aromas in every glass of champagne. PMID- 19805336 TI - Chemoenzymatic elaboration of monosaccharides using engineered cytochrome P450BM3 demethylases. AB - Polysaccharides comprise an extremely important class of biopolymers that play critical roles in a wide range of biological processes, but the synthesis of these compounds is challenging because of their complex structures. We have developed a chemoenzymatic method for regioselective deprotection of monosaccharide substrates using engineered Bacillus megaterium cytochrome P450 (P450(BM3)) demethylases that provides a highly efficient means to access valuable intermediates, which can be converted to a wide range of substituted monosaccharides and polysaccharides. Demethylases displaying high levels of regioselectivity toward a number of protected monosaccharides were identified using a combination of protein and substrate engineering, suggesting that this approach ultimately could be used in the synthesis of a wide range of substituted mono- and polysaccharides for studies in chemistry, biology, and medicine. PMID- 19805337 TI - Strong-field control and spectroscopy of attosecond electron-hole dynamics in molecules. AB - Molecular structures, dynamics and chemical properties are determined by shared electrons in valence shells. We show how one can selectively remove a valence electron from either Pi vs. Sigma or bonding vs. nonbonding orbital by applying an intense infrared laser field to an ensemble of aligned molecules. In molecules, such ionization often induces multielectron dynamics on the attosecond time scale. Ionizing laser field also allows one to record and reconstruct these dynamics with attosecond temporal and sub-Angstrom spatial resolution. Reconstruction relies on monitoring and controlling high-frequency emission produced when the liberated electron recombines with the valence shell hole created by ionization. PMID- 19805338 TI - A quantitative model for age-dependent expression of the p16INK4a tumor suppressor. AB - Recent work has shown that expression of the p16(INK4a) tumor suppressor increases with chronological age. Expression is accelerated by gerontogenic behaviors such as tobacco use and physical inactivity, and is also influenced by allelic genotype of a polymorphic single nucleotide polymorphism (SNP) rs10757278 that is physically linked with the p16(INK4a) ORF. To understand the relationship between p16(INK4a) expression, chronologic age, subject characteristics and host genetics, we sought to develop a mathematical model that links p16(INK4a) expression with aging. Using an annotated dataset of 170 healthy adults for whom p16(INK4a) expression and subject genotypes were known, we developed two alternative stochastic models that relate p16(INK4a) expression to age, smoking, exercise and rs10757278 genotype. Levels of p16(INK4a) increased exponentially and then saturated at later chronologic ages. The model, which best fit the data, suggests saturation occurs because of p16(INK4a)-dependent attrition of subjects at older chronologic ages, presumably due to death or chronic illness. An important feature of our model is that factors that contribute to death in a non p16(INK4a)-dependent manner do not affect our analysis. Interestingly, tobacco related increases in p16(INK4a) expression are predicted to arise from a decrease in the rate of p16(INK4a)-dependent death. This analysis is most consistent with the model that p16(INK4a) expression monotonically increases with age, and higher expression is associated with increased subject attrition. PMID- 19805339 TI - Physiological function and transplantation of scaffold-free and vascularized human cardiac muscle tissue. AB - Success of human myocardial tissue engineering for cardiac repair has been limited by adverse effects of scaffold materials, necrosis at the tissue core, and poor survival after transplantation due to ischemic injury. Here, we report the development of scaffold-free prevascularized human heart tissue that survives in vivo transplantation and integrates with the host coronary circulation. Human embryonic stem cells (hESCs) were differentiated to cardiomyocytes by using activin A and BMP-4 and then placed into suspension on a rotating orbital shaker to create human cardiac tissue patches. Optimization of patch culture medium significantly increased cardiomyocyte viability in patch centers. These patches, composed only of enriched cardiomyocytes, did not survive to form significant grafts after implantation in vivo. To test the hypothesis that ischemic injury after transplantation would be attenuated by accelerated angiogenesis, we created "second-generation," prevascularized, and entirely human patches from cardiomyocytes, endothelial cells (both human umbilical vein and hESC-derived endothelial cells), and fibroblasts. Functionally, vascularized patches actively contracted, could be electrically paced, and exhibited passive mechanics more similar to myocardium than patches comprising only cardiomyocytes. Implantation of these patches resulted in 10-fold larger cell grafts compared with patches composed only of cardiomyocytes. Moreover, the preformed human microvessels anastomosed with the rat host coronary circulation and delivered blood to the grafts. Thus, inclusion of vascular and stromal elements enhanced the in vitro performance of engineered human myocardium and markedly improved viability after transplantation. These studies demonstrate the importance of including vascular and stromal elements when designing human tissues for regenerative therapies. PMID- 19805340 TI - Survey of large protein complexes in D. vulgaris reveals great structural diversity. AB - An unbiased survey has been made of the stable, most abundant multi-protein complexes in Desulfovibrio vulgaris Hildenborough (DvH) that are larger than Mr approximately 400 k. The quaternary structures for 8 of the 16 complexes purified during this work were determined by single-particle reconstruction of negatively stained specimens, a success rate approximately 10 times greater than that of previous "proteomic" screens. In addition, the subunit compositions and stoichiometries of the remaining complexes were determined by biochemical methods. Our data show that the structures of only two of these large complexes, out of the 13 in this set that have recognizable functions, can be modeled with confidence based on the structures of known homologs. These results indicate that there is significantly greater variability in the way that homologous prokaryotic macromolecular complexes are assembled than has generally been appreciated. As a consequence, we suggest that relying solely on previously determined quaternary structures for homologous proteins may not be sufficient to properly understand their role in another cell of interest. PMID- 19805341 TI - Reconstitution of phospholipid translocase activity with purified Drs2p, a type IV P-type ATPase from budding yeast. AB - Type-IV P-type ATPases (P4-ATPases) are putative phospholipid translocases, or flippases, that translocate specific phospholipid substrates from the exofacial to the cytosolic leaflet of membranes to generate phospholipid asymmetry. In addition, the activity of Drs2p, a P4-ATPase from Saccharomyces cerevisiae, is required for vesicle-mediated protein transport from the Golgi and endosomes, suggesting a role for phospholipid translocation in vesicle budding. Drs2p is necessary for translocation of a fluorescent phosphatidylserine analogue across purified Golgi membranes. However, a flippase activity has not been reconstituted with purified Drs2p or any other P4-ATPase, so whether these ATPases directly pump phospholipid across the membrane bilayer is unknown. Here, we show that Drs2p can catalyze phospholipid translocation directly through purification and reconstitution of this P4-ATPase into proteoliposomes. The noncatalytic subunit, Cdc50p, also was reconstituted in the proteoliposome, although at a substoichiometric concentration relative to Drs2p. In proteoliposomes containing Drs2p, a phosphatidylserine analogue was actively flipped across the liposome bilayer to the outer leaflet in the presence of Mg(2+)-ATP, whereas no activity toward the phosphatidylcholine or sphingomyelin analogues was observed. This flippase activity was mediated by Drs2p, because protein-free liposomes or proteoliposomes reconstituted with a catalytically inactive form of Drs2p showed no translocation activity. These data demonstrate for the first time the reconstitution of a flippase activity with a purified P4-ATPase. PMID- 19805342 TI - GTP-induced conformational changes in septins and implications for function. AB - Septins constitute a group of GTP-binding proteins involved in cytokinesis and other essential cellular functions. They form heterooligomeric complexes that polymerize into nonpolar filaments and are dynamic during different stages of the cell cycle. Posttranslational modifications and interacting partners are widely accepted regulators of septin filament function, but the contribution of nucleotide is undefined due to a lack of detailed structural information. Previous low-resolution structures showed that the G domain assembles into a linear polymer with 2 different interfaces involving the N and C termini and the G binding sites. Here we report the crystal structure of SEPT2 bound to GppNHp at 2.9 A resolution. GTP binding induces conformational changes in the switch regions at the G interfaces, which are transmitted to the N-terminal helix and also affect the NC interface. Biochemical studies and sequence alignment suggest that a threonine, which is conserved in certain subgroups of septins, is responsible for GTP hydrolysis. Although this threonine is not present in yeast CDC3 and CDC11, its mutation in CDC10 and CDC12 induces temperature sensitivity. Highly conserved contact residues identified in the G interface are shown to be necessary for Cdc3-10, but not Cdc11-12, heterodimer formation and cell growth in yeast. Based on our findings, we propose that GTP binding/hydrolysis and the nature of the nucleotide influence the stability of interfaces in heterooligomeric and polymeric septins and are required for proper septin filament assembly/disassembly. These data also offer a first rationale for subdividing human septins into different functional subgroups. PMID- 19805343 TI - Repression of DNA-binding dependent glucocorticoid receptor-mediated gene expression. AB - The glucocorticoid receptor (GR) affects the transcription of genes involved in diverse processes, including energy metabolism and the immune response, through DNA-binding dependent and independent mechanisms. The DNA-binding dependent mechanism occurs by direct binding of GR to glucocorticoid response elements (GREs) at regulatory regions of target genes. The DNA-binding independent mechanism involves binding of GR to transcription factors and coactivators that, in turn, contact DNA. A small molecule that competes with GR for binding to GREs could be expected to affect the DNA-dependent pathway selectively by interfering with the protein-DNA interface. We show that a DNA-binding polyamide that targets the consensus GRE sequence binds the glucocorticoid-induced zipper (GILZ) GRE, inhibits expression of GILZ and several other known GR target genes, and reduces GR occupancy at the GILZ promoter. Genome-wide expression analysis of the effects of this polyamide on a set of glucocorticoid-induced and -repressed genes could help to elucidate the mechanism of GR regulation for these genes. PMID- 19805344 TI - Structure of apo-CAP reveals that large conformational changes are necessary for DNA binding. AB - The binding of cAMP to the Escherichia coli catabolite gene activator protein (CAP) produces a conformational change that enables it to bind specific DNA sequences and regulate transcription, which it cannot do in the absence of the nucleotide. The crystal structures of the unliganded CAP containing a D138L mutation and the unliganded WT CAP were determined at 2.3 and 3.6 A resolution, respectively, and reveal that the two DNA binding domains have dimerized into one rigid body and their two DNA recognition helices become buried. The WT structure shows multiple orientations of this rigid body relative to the nucleotide binding domain supporting earlier biochemical data suggesting that the inactive form exists in an equilibrium among different conformations. Comparison of the structures of the liganded and unliganded CAP suggests that cAMP stabilizes the active DNA binding conformation of CAP through the interactions that the N(6) of the adenosine makes with the C-helices. These interactions are associated with the reorientation and elongation of the C-helices that precludes the formation of the inactive structure. PMID- 19805345 TI - Barrier-to-autointegration factor (BAF) condenses DNA by looping. AB - Barrier-to-autointegration factor (BAF) is a protein that has been proposed to compact retroviral DNA, making it inaccessible as a target for self-destructive integration into itself (autointegration). BAF also plays an important role in nuclear organization. We studied the mechanism of DNA condensation by BAF using total internal reflection fluorescence microscopy. We found that BAF compacts DNA by a looping mechanism. Dissociation of BAF from DNA occurs with multiphasic kinetics; an initial fast phase is followed by a much slower dissociation phase. The mechanistic basis of the broad timescale of dissociation is discussed. This behavior mimics the dissociation of BAF from retroviral DNA within preintegration complexes as monitored by functional assays. Thus the DNA binding properties of BAF may alone be sufficient to account for its association with the preintegration complex. PMID- 19805346 TI - A progenitor cell origin of myeloid malignancies. AB - All cancers rely on cells that have properties of long-term self-renewal or "stemness" to maintain and propagate the tumor, but the cell of origin of most cancers is still unknown. Here, we design a stochastic mathematical model of hematopoietic stem and progenitor cells to study the evolutionary dynamics of cancer initiation. We consider different evolutionary pathways leading to cancer initiating cells in JAK2V617F-positive myeloproliferative neoplasms (MPN): (i) the JAK2V617F mutation may arise in a stem cell; (ii) a progenitor cell may first acquire a mutation conferring self-renewal, followed by acquisition of the JAK2V617F mutation; (iii) the JAK2V617F mutation may first emerge in a progenitor cell, followed by a mutation conferring self-renewal; and (iv) a mutation conferring self-renewal to progenitors may arise in the stem cell population without causing a change in the stem cell's phenotype, followed by the JAK2V617F mutation emerging in a progenitor cell. We find mathematical evidence that a progenitor is the most likely cell of origin of JAK2V617F-mutant MPN. These results may also have relevance to other tumor types arising in tissues that are organized as a differentiation hierarchy. PMID- 19805347 TI - Fast screening of protein surfaces using geometric invariant fingerprints. AB - We develop a rapid and efficient method for the comparison of protein local surface similarities using geometric invariants (fingerprints). By combining fast fingerprint comparison with explicit alignment, we successfully screen the entire Protein Data Bank for proteins that possess local surface similarities. Our method is independent of sequence and fold similarities, and has potential application to protein structure annotation and protein-protein interface design. PMID- 19805348 TI - Tetramer opening in LacI-mediated DNA looping. AB - Lactose repressor protein (LacI) controls transcription of the genes involved in lactose metabolism in bacteria. Essential to optimal LacI-mediated regulation is its ability to bind simultaneously to two operators, forming a loop on the intervening DNA. Recently, several lines of evidence (both theoretical and experimental) have suggested various possible loop structures associated with different DNA binding topologies and LacI tetramer structural conformations (adopted by flexing about the C-terminal tetramerization domain). We address, specifically, the role of protein opening in loop formation by employing the single-molecule tethered particle motion method on LacI protein mutants chemically cross-linked at different positions along the cleft between the two dimers. Measurements on the wild-type and uncross-linked LacI mutants led to the observation of two distinct levels of short tether length, associated with two different DNA looping structures. Restricting conformational flexibility of the protein by chemical cross-linking induces pronounced effects. Crosslinking the dimers at the level of the N-terminal DNA binding head (E36C) completely suppresses looping, whereas cross-linking near the C-terminal tetramerization domain (Q231C) results in changes of looping geometry detected by the measured tether length distributions. These observations lead to the conclusion that tetramer opening plays a definite role in at least a subset of LacI/DNA loop conformations. PMID- 19805349 TI - No facilitator required for membrane transport of hydrogen sulfide. AB - Hydrogen sulfide (H(2)S) has emerged as a new and important member in the group of gaseous signaling molecules. However, the molecular transport mechanism has not yet been identified. Because of structural similarities with H(2)O, it was hypothesized that aquaporins may facilitate H(2)S transport across cell membranes. We tested this hypothesis by reconstituting the archeal aquaporin AfAQP from sulfide reducing bacteria Archaeoglobus fulgidus into planar membranes and by monitoring the resulting facilitation of osmotic water flow and H(2)S flux. To measure H(2)O and H(2)S fluxes, respectively, sodium ion dilution and buffer acidification by proton release (H(2)S left arrow over right arrow H(+) + HS(-)) were recorded in the immediate membrane vicinity. Both sodium ion concentration and pH were measured by scanning ion-selective microelectrodes. A lower limit of lipid bilayer permeability to H(2)S, P(M,H(2)S) >or = 0.5 +/- 0.4 cm/s was calculated by numerically solving the complete system of differential reaction diffusion equations and fitting the theoretical pH distribution to experimental pH profiles. Even though reconstitution of AfAQP significantly increased water permeability through planar lipid bilayers, P(M,H(2)S) remained unchanged. These results indicate that lipid membranes may well act as a barrier to water transport although they do not oppose a significant resistance to H(2)S diffusion. The fact that cholesterol and sphingomyelin reconstitution did not turn these membranes into an H(2)S barrier indicates that H(2)S transport through epithelial barriers, endothelial barriers, and membrane rafts also occurs by simple diffusion and does not require facilitation by membrane channels. PMID- 19805350 TI - Membrane structure and conformational changes of the antibiotic heterodimeric peptide distinctin by solid-state NMR spectroscopy. AB - The heterodimeric antimicrobial peptide distinctin is composed of 2 linear peptide chains of 22- and 25-aa residues that are connected by a single intermolecular S-S bond. This heterodimer has been considered to be a unique example of a previously unrecorded class of bioactive peptides. Here the 2 distinctin chains were prepared by chemical peptide synthesis in quantitative amounts and labeled with (15)N, as well as (15)N and (2)H, at selected residues, respectively, and the heterodimer was formed by oxidation. CD spectroscopy indicates a high content of helical secondary structures when associated with POPC/POPG 3:1 vesicles or in membrane-mimetic environments. The propensity for helix formation follows the order heterodimer >chain 2 >chain 1, suggesting that peptide-peptide and peptide-lipid interactions both help in stabilizing this secondary structure. In a subsequent step the peptides were reconstituted into oriented phospholipid bilayers and investigated by (2)H and proton-decoupled (15)N solid-state NMR spectroscopy. Whereas chain 2 stably inserts into the membrane at orientations close to perfectly parallel to the membrane surface in the presence or absence of chain 1, the latter adopts a more tilted alignment, which further increases in the heterodimer. The data suggest that membrane interactions result in considerable conformational rearrangements of the heterodimer. Therefore, chain 2 stably anchors the heterodimer in the membrane, whereas chain 1 interacts more loosely with the bilayer. These structural observations are consistent with the antimicrobial activities when the individual chains are compared to the dimer. PMID- 19805351 TI - Order of lipid phases in model and plasma membranes. AB - Lipid rafts are nanoscopic assemblies of sphingolipids, cholesterol, and specific membrane proteins that contribute to lateral heterogeneity in eukaryotic membranes. Separation of artificial membranes into liquid-ordered (Lo) and liquid disordered phases is regarded as a common model for this compartmentalization. However, tight lipid packing in Lo phases seems to conflict with efficient partitioning of raft-associated transmembrane (TM) proteins. To assess membrane order as a component of raft organization, we performed fluorescence spectroscopy and microscopy with the membrane probes Laurdan and C-laurdan. First, we assessed lipid packing in model membranes of various compositions and found cholesterol and acyl chain dependence of membrane order. Then we probed cell membranes by using two novel systems that exhibit inducible phase separation: giant plasma membrane vesicles [Baumgart et al. (2007) Proc Natl Acad Sci USA 104:3165-3170] and plasma membrane spheres. Notably, only the latter support selective inclusion of raft TM proteins with the ganglioside GM1 into one phase. We measured comparable small differences in order between the separated phases of both biomembranes. Lateral packing in the ordered phase of giant plasma membrane vesicles resembled the Lo domain of model membranes, whereas the GM1 phase in plasma membrane spheres exhibited considerably lower order, consistent with different partitioning of lipid and TM protein markers. Thus, lipid-mediated coalescence of the GM1 raft domain seems to be distinct from the formation of a Lo phase, suggesting additional interactions between proteins and lipids to be effective. PMID- 19805352 TI - Macromolecular crowding induced elongation and compaction of single DNA molecules confined in a nanochannel. AB - The effect of dextran nanoparticles on the conformation and compaction of single DNA molecules confined in a nanochannel was investigated with fluorescence microscopy. It was observed that the DNA molecules elongate and eventually condense into a compact form with increasing volume fraction of the crowding agent. Under crowded conditions, the channel diameter is effectively reduced, which is interpreted in terms of depletion in DNA segment density in the interfacial region next to the channel wall. Confinement in a nanochannel also facilitates compaction with a neutral crowding agent at low ionic strength. The threshold volume fraction for condensation is proportional to the size of the nanoparticle, due to depletion induced attraction between DNA segments. We found that the effect of crowding is not only related to the colligative properties of the agent and that confinement is also important. It is the interplay between anisotropic confinement and osmotic pressure which gives the elongated conformation and the possibility for condensation at low ionic strength. PMID- 19805353 TI - Function of IRE1 alpha in the placenta is essential for placental development and embryonic viability. AB - Inositol requiring enzyme-1 (IRE1), a protein located on the endoplasmic reticulum (ER) membrane, is highly conserved from yeast to humans. This protein is activated during ER stress and induces cellular adaptive responses to the stress. In mice, IRE1alpha inactivation results in widespread developmental defects, leading to embryonic death after 12.5 days of gestation. However, the cause of this embryonic lethality is not fully understood. Here, by using in vivo imaging analysis and conventional knockout mice, respectively, we showed that IRE1alpha was activated predominantly in the placenta and that loss of IRE1alpha led to reduction in vascular endothelial growth factor-A and severe dysfunction of the labyrinth in the placenta, a highly developed tissue of blood vessels. We also used a conditional knockout strategy to demonstrate that IRE1alpha-deficient embryos supplied with functionally normal placentas can be born alive. Fetal liver hypoplasia thought to be responsible for the embryonic lethality of IRE1alpha-null mice was virtually absent in rescued IRE1alpha-null pups. These findings reveal that IRE1alpha plays an essential function in extraembryonic tissues and highlight the relationship of physiological ER stress and angiogenesis in the placenta during pregnancy in mammals. PMID- 19805354 TI - Sulforaphane destabilizes the androgen receptor in prostate cancer cells by inactivating histone deacetylase 6. AB - High consumption of cruciferous vegetables is associated with a reduced risk of prostate cancer in epidemiological studies. There is preliminary evidence that sulforaphane, derived from glucoraphanin found in a number of crucifers, may prevent and induce regression of prostate cancer and other malignancies in preclinical models, but the mechanisms that may explain these effects are not fully defined. Recent reports show that sulforaphane may impair prostate cancer growth through inhibition of histone deacetylases, which are up-regulated in cancer. Indeed, one of these enzymes, histone deacetylase 6 (HDAC6), influences the acetylation state of a key androgen receptor (AR) chaperone, HSP90. AR is the central signaling pathway in prostate cancer, and its inhibition is used for both prevention and treatment of this disease. However, it is not known whether the effects of sulforaphane involve suppression of AR. We hypothesized that sulforaphane treatment would lead to hyperacetylation of HSP90 and that this would destabilize AR and attenuate AR signaling. We confirmed this by demonstrating that sulforaphane enhances HSP90 acetylation, thereby inhibiting its association with AR. Moreover, AR is subsequently degraded in the proteasome, which leads to reduced AR target gene expression and reduced AR occupancy at its target genes. Finally, sulforaphane inhibits HDAC6 deacetylase activity, and the effects of sulforaphane on AR protein are abrogated by overexpression of HDAC6 and mimicked by HDAC6 siRNA. The inactivation by sulforaphane of HDAC6-mediated HSP90 deacetylation and consequent attenuation of AR signaling represents a newly defined mechanism that may help explain this agent's effects in prostate cancer. PMID- 19805355 TI - Dual role of K ATP channel C-terminal motif in membrane targeting and metabolic regulation. AB - The coordinated sorting of ion channels to specific plasma membrane domains is necessary for excitable cell physiology. K(ATP) channels, assembled from pore forming (Kir6.x) and regulatory sulfonylurea receptor subunits, are critical electrical transducers of the metabolic state of excitable tissues, including skeletal and smooth muscle, heart, brain, kidney, and pancreas. Here we show that the C-terminal domain of Kir6.2 contains a motif conferring membrane targeting in primary excitable cells. Kir6.2 lacking this motif displays aberrant channel targeting due to loss of association with the membrane adapter ankyrin-B (AnkB). Moreover, we demonstrate that this Kir6.2 C-terminal AnkB-binding motif (ABM) serves a dual role in K(ATP) channel trafficking and membrane metabolic regulation and dysfunction in these pathways results in human excitable cell disease. Thus, the K(ATP) channel ABM serves as a previously unrecognized bifunctional touch-point for grading K(ATP) channel gating and membrane targeting and may play a fundamental role in controlling excitable cell metabolic regulation. PMID- 19805356 TI - IGFBP2 is a candidate biomarker for Ink4a-Arf status and a therapeutic target for high-grade gliomas. AB - The levels of insulin-like growth factor-binding protein 2 (IGFBP2) are elevated during progression of many human cancers. By using a glial-specific transgenic mouse system (RCAS/Ntv-a), we reported previously that IGFBP2 is an oncogenic factor for glioma progression in combination with platelet-derived growth factor beta (PDGFB). Because the INK4a-ARF locus is often deleted in high-grade gliomas (anaplastic oligodendroglioma and glioblastoma), we investigated the effect of the Ink4a-Arf-null background on IGFBP2-mediated progression of PDGFB-initiated oligodendroglioma. We demonstrate here that homozygous deletion of Ink4a-Arf bypasses the requirement of exogenously introduced IGFBP2 for glioma progression. Instead, absence of Ink4a-Arf resulted in elevated endogenous tumor cell IGFBP2. An inverse relationship between p16(INK4a) and IGFBP2 expression was also observed in human glioma tissue samples and in 90 different cancer cell lines by using Western blotting and reverse-phase protein lysate arrays. When endogenous IGFBP2 expression was attenuated by an RCAS vector expressing antisense IGFBP2 in our mouse model, a decreased incidence of anaplastic oligodendroglioma as well as prolonged survival was observed. Thus, p16(INK4a) is a negative regulator of the IGFBP2 oncogene. Loss of Ink4a-Arf results in increased IGFBP2, which contributes to glioma progression, thereby implicating IGFBP2 as a marker and potential therapeutic target for Ink4a-Arf-deleted gliomas. PMID- 19805357 TI - Functional interaction between native G protein-coupled purinergic receptors in Xenopus follicles. AB - Purinergic receptors are expressed in the membrane of the follicular cell layer that communicates with the Xenopus oocyte. Adenosine (Ado) generates a cAMP dependent K(+) current (I(K,cAMP)), whereas ATP activates a Cl(-) current (F(Cl)) and has a dual effect on I(K,cAMP), provoking both its activation and inhibition. Here, purinergic responses were studied electrophysiologically, first in the whole follicle (w.f.), and then in the same follicle after removal of its epithelium/theca layers (e.t.r. follicle). Responses were analyzed as the ratio of the current amplitudes (i(etr)/i(wf)) in the two preparations. For ATP activation of I(K,cAMP) and F(Cl), the ratios i(etr)/i(wf) were 0.053 and 22, respectively, whereas that for Ado was 0.75. Thus, epithelium/theca removal drastically altered the ATP response, suggesting a change in the signaling pathway that correlated with changes in the pharmacological characteristics: the half-maximal effective concentration for activation of the main current in w.f. (I(K,cAMP)) was 14 +/- 3.8 microM [Hill coefficient (nH) = 2.7 +/- 0.61], and that in e.t.r. follicles (F(Cl)) was 1.8 +/- 0.68 microM (nH = 0.76 +/- 0.09), whereas Ado-response parameters did not change. Responses to UTP and beta,gamma methylene-ATP, specific agonists for I(K,cAMP) inhibition and activation, respectively, indicated that in e.t.r. follicles inhibition increased and activation decreased drastically. Thus, purinergic responses were not independent; instead, they were functionally linked. We hypothesize that this property was due to direct interactions between receptors for Ado (A2 subtype) and ATP (P2Y subtype) in the Xenopus follicle. PMID- 19805358 TI - Ubiquitination of histone H2B regulates chromatin dynamics by enhancing nucleosome stability. AB - The mechanism by which ubiquitination of histone H2B (H2Bub1) regulates H3-K4 and -K79 methylation and the histone H2A-H2B chaperone Spt16-mediated nucleosome dynamics during transcription is not fully understood. Upon investigating the effect of H2Bub1 on chromatin structure, we find that contrary to the supposed role for H2Bub1 in opening up chromatin, it is important for nucleosome stability. First, we show that H2Bub1 does not function as a "wedge" to non specifically unfold chromatin, as replacement of ubiquitin with a bulkier SUMO molecule conjugated to the C-terminal helix of H2B cannot functionally support H3 K4 and -K79 methylation. Second, using a series of biochemical analyses, we demonstrate that nucleosome stability is reduced or enhanced, when the levels of H2Bub1 are abolished or increased, respectively. Besides transcription elongation, we show that H2Bub1 regulates initiation by stabilizing nucleosomes positioned over the promoters of repressed genes. Collectively, our study reveals an intrinsic difference in the property of chromatin assembled in the presence or absence of H2Bub1 and implicates the regulation of nucleosome stability as the mechanism by which H2Bub1 modulates nucleosome dynamics and histone methylation during transcription. PMID- 19805359 TI - Central role of TRAF-interacting protein in a new model of brain sexual differentiation. AB - Sexually dimorphic brain nuclei underlie gender-specific neural functions and susceptibility to disease, but the developmental basis of dimorphisms is poorly understood. In these studies, we focused on the anteroventral periventricular nucleus (AVPV), a nucleus that is larger in females and critical for the female typical cyclic surge pattern of luteinizing hormone (LH) release. Sex differences in the size and function of the AVPV result from apoptosis that occurs preferentially in the developing male. To identify upstream pathways responsible for sexual differentiation of the AVPV, we used targeted apoptosis microarrays and in vivo and in vitro follow-up studies. We found that the tumor necrosis factor alpha (TNFalpha)-TNF receptor 2 (TNFR2)-NFkappaB cell survival pathway is active in postnatal day 2 (PND2) female AVPV and repressed in male counterparts. Genes encoding key members of this pathway were expressed exclusively in GABAergic neurons. One gene in particular, TNF receptor-associated factor 2 (TRAF2)-inhibiting protein (trip), was higher in males and it inhibited both TNFalpha-dependent NFkappaB activation and bcl-2 gene expression. The male AVPV also had higher levels of bax and bad mRNA, but neither of these genes was regulated by either TNFalpha or TRIP. Finally, the trip gene was not expressed in the sexually dimorphic nucleus of the preoptic area (SDN-POA), a nucleus in which apoptosis is higher in females than males. These findings form the basis of a new model of sexual differentiation of the AVPV that may also apply to the development of other sexually dimorphic nuclei. PMID- 19805360 TI - The tyrosine phosphatase Shp2 (PTPN11) directs Neuregulin-1/ErbB signaling throughout Schwann cell development. AB - The nonreceptor tyrosine phosphatase Shp2 (PTPN11) has been implicated in tyrosine kinase, cytokine, and integrin receptor signaling. We show here that conditional mutation of Shp2 in neural crest cells and in myelinating Schwann cells resulted in deficits in glial development that are remarkably similar to those observed in mice mutant for Neuregulin-1 (Nrg1) or the Nrg1 receptors, ErbB2 and ErbB3. In cultured Shp2 mutant Schwann cells, Nrg1-evoked cellular responses like proliferation and migration were virtually abolished, and Nrg1 dependent intracellular signaling was altered. Pharmacological inhibition of Src family kinases mimicked all cellular and biochemical effects of the Shp2 mutation, implicating Src as a primary Shp2 target during Nrg1 signaling. Together, our genetic and biochemical analyses demonstrate that Shp2 is an essential component in the transduction of Nrg1/ErbB signals. PMID- 19805361 TI - Loss of Erk3 function in mice leads to intrauterine growth restriction, pulmonary immaturity, and neonatal lethality. AB - Extracellular signal-regulated kinase 3 (Erk3) is an atypical member of the mitogen-activated protein (MAP) kinase family. No function has yet been ascribed to this MAP kinase. Here we show that targeted disruption of the Mapk6 gene (encoding Erk3) leads to intrauterine growth restriction, associated with marked pulmonary hypoplasia, and early neonatal death during the first day of life. Around 40% of Erk3(-/-) neonates die within minutes after birth from acute respiratory failure. Erk3-deficient mice have normal lung-branching morphogenesis, but show delayed lung maturation characterized by decreased sacculation, atelectasis, and defective type II pneumocyte differentiation. Interestingly, in utero administration of glucocorticoid promoted fetal lung maturity and rescued differentiation of type II cells, but failed to alter the neonatal lethality. We observed that loss of Erk3 retards intrauterine growth, as reflected by a marked reduction in fetal lung, heart, and liver weights, and by low body weight at birth. Importantly, we found that insulin-like growth factor (IGF)-2 levels are decreased in the serum of Erk3-deficient mice. Our findings reveal a critical role for Erk3 in the establishment of fetal growth potential and pulmonary function in the mouse. PMID- 19805362 TI - Identifying sub-optimal responses to ivermectin in the treatment of River Blindness. AB - Identification of drug resistance before it becomes a public health concern requires a clear distinction between what constitutes a normal and a suboptimal treatment response. A novel method of analyzing drug efficacy studies in human helminthiases is proposed and used to investigate recent claims of atypical responses to ivermectin in the treatment of River Blindness. The variability in the rate at which Onchocerca volvulus microfilariae repopulate host's skin following ivermectin treatment is quantified using an individual-based onchocerciasis mathematical model. The model estimates a single skin repopulation rate for every host sampled, allowing reports of suboptimal responses to be statistically compared with responses from populations with no prior exposure to ivermectin. Statistically faster rates of skin repopulation were observed in 3 Ghanaian villages (treated 12-17 times), despite the wide variability in repopulation rates observed in ivermectin-naive populations. Another village previously thought to have high rates of skin repopulation was shown to be indistinguishable from the normal treatment response. The model is used to generate testable hypotheses to identify whether atypical rates of skin repopulation by microfilariae could result from low treatment coverage alone or provide evidence of decreased ivermectin efficacy. Further work linking phenotypic poor responses to treatment with parasite molecular genetics markers will be required to confirm drug resistance. Limitations of the skin-snipping method for estimating parasite load indicates that changes in the distribution of microfilarial repopulation rates, rather than their absolute values, maybe a more sensitive indicator of emerging ivermectin resistance. PMID- 19805363 TI - Fossil and molecular evidence constrain scenarios for the early evolutionary and biogeographic history of hystricognathous rodents. AB - The early evolutionary and paleobiogeographic history of the diverse rodent clade Hystricognathi, which contains Hystricidae (Old World porcupines), Caviomorpha (the endemic South American rodents), and African Phiomorpha (cane rats, dassie rats, and blesmols) is of great interest to students of mammalian evolution, but remains poorly understood because of a poor early fossil record. Here we describe the oldest well-dated hystricognathous rodents from an earliest late Eocene (approximately 37 Ma) fossil locality in the Fayum Depression of northern Egypt. These taxa exhibit a combination of primitive and derived features, the former shared with Asian "baluchimyine" rodents, and the latter shared with Oligocene phiomorphs and caviomorphs. Phylogenetic analysis incorporating morphological, temporal, geographic, and molecular information places the new taxa as successive sister groups of crown Hystricognathi, and supports an Asian origin for stem Hystricognathi and an Afro-Arabian origin for crown Hystricognathi, stem Hystricidae, and stem Caviomorpha. Molecular dating of early divergences within Hystricognathi, using a Bayesian "relaxed clock" approach and multiple fossil calibrations, suggests that the split between Hystricidae and the phiomorph caviomorph clade occurred approximately 39 Ma, and that phiomorphs and caviomorphs diverged approximately 36 Ma. These results are remarkably congruent with our phylogenetic results and the fossil record of hystricognathous rodent evolution in Afro-Arabia and South America. PMID- 19805364 TI - Fine-scale mergers of chloroplast and mitochondrial genes create functional, transcompartmentally chimeric mitochondrial genes. AB - The mitochondrial genomes of flowering plants possess a promiscuous proclivity for taking up sequences from the chloroplast genome. All characterized chloroplast integrants exist apart from native mitochondrial genes, and only a few, involving chloroplast tRNA genes that have functionally supplanted their mitochondrial counterparts, appear to be of functional consequence. We developed a novel computational approach to search for homologous recombination (gene conversion) in a large number of sequences and applied it to 22 mitochondrial and chloroplast gene pairs, which last shared common ancestry some 2 billion years ago. We found evidence of recurrent conversion of short patches of mitochondrial genes by chloroplast homologs during angiosperm evolution, but no evidence of gene conversion in the opposite direction. All 9 putative conversion events involve the atp1/atpA gene encoding the alpha subunit of ATP synthase, which is unusually well conserved between the 2 organelles and the only shared gene that is widely sequenced across plant mitochondria. Moreover, all conversions were limited to the 2 regions of greatest nucleotide and amino acid conservation of atp1/atpA. These observations probably reflect constraints operating on both the occurrence and fixation of recombination between ancient homologs. These findings indicate that recombination between anciently related sequences is more frequent than previously appreciated and creates functional mitochondrial genes of chimeric origin. These results also have implications for the widespread use of mitochondrial atp1 in phylogeny reconstruction. PMID- 19805365 TI - Mediator complex association with constitutively transcribed genes in yeast. AB - Mediator is a large, multisubunit complex that is essential for transcription of mRNA by RNA Pol II in eukaryotes and is believed to bridge transcriptional activators and the general transcription machinery. However, several recent studies suggest that the requirement for Mediator during transcriptional activation is not universal, but rather activator dependent, and may be indirect for some genes. Here we have investigated Mediator association with several constitutively transcribed genes in yeast by comparing a yeast strain that harbors a temperature-sensitive mutation in an essential Mediator subunit, Srb4, with its wild-type (WT) counterpart. We find modest association of Mediator with constitutively active genes and show that this association is strongly decreased in srb4 ts yeast, whereas association with a nontranscribed region or repressed gene promoters is lower and unaffected in the mutant yeast. The tail module of Mediator remains associated with ribosomal protein (RP) gene promoters in srb4 ts yeast, while subunits from the head and middle modules are lost. Tail module association at Rap1-dependent gene promoters is lost in rap1 ts yeast, indicating that Rap1 is required for Mediator recruitment at these gene promoters and that its recruitment occurs via the tail module. Pol II association is also rapidly and severely affected in srb4 ts yeast, indicating that Mediator is directly required for pol II association at constitutively transcribed genes. Our results are consistent with Mediator functioning as a general transcription factor in yeast. PMID- 19805366 TI - BG1 has a major role in MHC-linked resistance to malignant lymphoma in the chicken. AB - Pathogen selection is postulated to drive MHC allelic diversity at loci for antigen presentation. However, readily apparent MHC infectious disease associations are rare in most species. The strong link between MHC-B haplotype and the occurrence of virally induced tumors in the chicken provides a means for defining the relationship between pathogen selection and MHC polymorphism. Here, we verified a significant difference in resistance to gallid herpesvirus-2 (GaHV 2)-induced lymphomas (Marek's disease) conferred by two closely-related recombinant MHC-B haplotypes. We mapped the crossover breakpoints that distinguish these haplotypes to the highly polymorphic BG1 locus. BG1 encodes an Ig-superfamily type I transmembrane receptor-like protein that contains an immunoreceptor tyrosine-based inhibition motif (ITIM), which undergoes phosphorylation and is recognized by Src homology 2 domain-containing protein tyrosine phosphatase (SHP-2). The recombinant haplotypes are identical, except for differences within the BG1 3'-untranslated region (3'-UTR). The 3'-UTR of the BG1 allele associated with increased lymphoma contains a 225-bp insert of retroviral origin and showed greater inhibition of luciferase reporter gene translation compared to the other allele. These findings suggest that BG1 could affect the outcome of GaHV-2 infection through modulation of the lymphoid cell responsiveness to infection, a condition that is critical for GaHV-2 replication and in which the MHC-B haplotype has been previously implicated. This work provides a mechanism by which MHC-B region genetics contributes to the incidence of GaHV-2-induced malignant lymphoma in the chicken and invites consideration of the possibility that similar mechanisms might affect the incidence of lymphomas associated with other oncogenic viral infections. PMID- 19805367 TI - Elucidating the genetic architecture of familial schizophrenia using rare copy number variant and linkage scans. AB - To elucidate the genetic architecture of familial schizophrenia we combine linkage analysis with studies of fine-level chromosomal variation in families recruited from the Afrikaner population in South Africa. We demonstrate that individually rare inherited copy number variants (CNVs) are more frequent in cases with familial schizophrenia as compared to unaffected controls and affect almost exclusively genic regions. Interestingly, we find that while the prevalence of rare structural variants is similar in familial and sporadic cases, the type of variants is markedly different. In addition, using a high-density linkage scan with a panel of nearly 2,000 markers, we identify a region on chromosome 13q34 that shows genome-wide significant linkage to schizophrenia and show that in the families not linked to this locus, there is evidence for linkage to chromosome 1p36. No causative CNVs were identified in either locus. Overall, our results from approaches designed to detect risk variants with relatively low frequency and high penetrance in a well-defined and relatively homogeneous population, provide strong empirical evidence supporting the notion that multiple genetic variants, including individually rare ones, that affect many different genes contribute to the genetic risk of familial schizophrenia. They also highlight differences in the genetic architecture of the familial and sporadic forms of the disease. PMID- 19805368 TI - Bioinformatics construction of the human cell surfaceome. AB - Cell surface proteins are excellent targets for diagnostic and therapeutic interventions. By using bioinformatics tools, we generated a catalog of 3,702 transmembrane proteins located at the surface of human cells (human cell surfaceome). We explored the genetic diversity of the human cell surfaceome at different levels, including the distribution of polymorphisms, conservation among eukaryotic species, and patterns of gene expression. By integrating expression information from a variety of sources, we were able to identify surfaceome genes with a restricted expression in normal tissues and/or differential expression in tumors, important characteristics for putative tumor targets. A high-throughput and efficient quantitative real-time PCR approach was used to validate 593 surfaceome genes selected on the basis of their expression pattern in normal and tumor samples. A number of candidates were identified as potential diagnostic and therapeutic targets for colorectal tumors and glioblastoma. Several candidate genes were also identified as coding for cell surface cancer/testis antigens. The human cell surfaceome will serve as a reference for further studies aimed at characterizing tumor targets at the surface of human cells. PMID- 19805369 TI - The same systemic autoimmune disease provokes arthritis and endocarditis via distinct mechanisms. AB - The immune mechanisms that provoke concomitant inflammation of synovial joints and cardiac valves in disorders such as rheumatic fever and systemic lupus erythematosus remain poorly defined. Here, we report the discovery of spontaneous endocarditis-in addition to their well-studied autoimmune arthritis-in K/BxN T cell receptor (TCR) transgenic mice. The same adaptive immune system elements were required for initiation of arthritis and endocarditis, and both diseases were dependent on autoantibodies. In contrast, the participation of key innate immune system molecules and perhaps T cells as effectors of inflammation differed between the 2 target tissues. Arthritis in K/BxN TCR transgenic mice depended primarily on complement C5 and not FcRgamma-using receptors; conversely, endocarditis depended essentially on FcRgamma receptors and not C5. Elucidating how a single systemic autoimmune disease engages distinct immune effector pathways to damage different target tissues is essential for optimizing the treatment of such disorders. PMID- 19805370 TI - 25-Hydroxycholesterol secreted by macrophages in response to Toll-like receptor activation suppresses immunoglobulin A production. AB - 25-Hydroxycholesterol is produced in mammalian tissues. The function of this oxysterol is unknown. Here we describe a central role for 25-hydroxycholesterol in regulating the immune system. In initial experiments, we found that stimulation of macrophage Toll-like receptors (TLR) induced expression of cholesterol 25-hydroxylase and the synthesis of 25-hydroxycholesterol. Treatment of naive B cells with nanomolar concentrations of 25-hydroxycholesterol suppressed IL-2-mediated stimulation of B cell proliferation, repressed activation-induced cytidine deaminase (AID) expression, and blocked class switch recombination, leading to markedly decreased IgA production. Consistent with these findings, deletion of the mouse cholesterol 25-hydroxylase gene caused an increase in serum IgA. Conversely, inactivation of the CYP7B1 oxysterol 7alpha hydroxylase, which degrades 25-hydroxycholesterol, decreased serum IgA. The suppression of IgA class switching in B cells by a macrophage-derived sterol in response to TLR activation provides a mechanism for local and systemic negative regulation of the adaptive immune response by the innate immune system. PMID- 19805371 TI - E3 ubiquitin ligase GRAIL controls primary T cell activation and oral tolerance. AB - T cell unresponsiveness or anergy is one of the mechanisms that maintain inactivity of self-reactive lymphocytes. E3 ubiquitin ligases are important mediators of the anergic state. The RING finger E3 ligase GRAIL is thought to selectively function in anergic T cells but its mechanism of action and its role in vivo are largely unknown. We show here that genetic deletion of Grail in mice leads not only to loss of an anergic phenotype in various models but also to hyperactivation of primary CD4(+) T cells. Grail(-/-) CD4(+) T cells hyperproliferate in vitro to TCR stimulation alone or with concomitant anti-CD28 costimulation, with transient increased survival. In vitro differentiated T helper 1 cells show slight but significant hypersecretion of IFN-gamma in Grail( /-) mice whereas Th2 and Th17 cytokine secretions are unchanged. Consistent with defective in vitro anergy, oral tolerance is abolished in vivo in OT-II TCR transgenic Grail(-/-) mice fed with ovalbumin. In experimental allergic encephalitis, a model of organ-specific autoimmunity, oral tolerization with myelin basic protein was abrogated as well in Grail(-/-) mice. On the protein level, Grail(-/-) naive T cells show no significant differences of total and phosphorylated levels of ZAP70, phospholipase Cgamma1, and MAP kinases p38 and JNK but elevated baseline levels of MAP kinase ERK1/2. In summary, we define a role for GRAIL in primary T cell activation, survival, and differentiation. In addition, we formally prove an indispensable role for GRAIL in T cell anergy and oral tolerance-a promising, antigen-specific strategy to treat autoimmune diseases. PMID- 19805372 TI - TSLP production by epithelial cells exposed to immunodeficiency virus triggers DC mediated mucosal infection of CD4+ T cells. AB - Mucosal dendritic cells have been implicated in the capture, storage, and transmission of HIV to CD4(+) T cells as well as in the promotion of HIV replication in activated CD4(+) T cells during the cognate T-cell and DC interaction. We report that HIV induces human genital mucosal epithelial cells to produce thymic stromal lymphopoietin (TSLP) via activation of the NFkappaB signaling pathway. The TSLP secreted by HIV exposed epithelial cells activated DC, which promoted proliferation and HIV-1 replication of co-cultured autologous CD4(+) T cells. In rhesus macaques, we observed dramatic increases in TSLP expression concurrent with an increase in viral replication in the vaginal tissues within the first 2 weeks after vaginal SIV exposure. These data suggest that HIV-mediated TSLP production by mucosal epithelial cells is a critical trigger for DC-mediated amplification of HIV-infection in activated CD4(+) T cells. The cross talk between mucosal epithelial cells and DC, mediated by HIV induced TSLP, may be an important mechanism for the high rate of HIV infection in women through the vaginal mucosa. PMID- 19805373 TI - A mathematical model of ischemic cutaneous wounds. AB - Chronic wounds represent a major public health problem affecting 6.5 million people in the United States. Ischemia, primarily caused by peripheral artery diseases, represents a major complicating factor in cutaneous wound healing. In this work, we sought to develop a mathematical model of ischemic dermal wounds. The model consists of a coupled system of partial differential equations in the partially healed region, with the wound boundary as a free boundary. The extracellular matrix (ECM) is assumed to be viscoelastic, and the free boundary moves with the velocity of the ECM at the boundary. The model equations involve the concentration of oxygen, PDGF and VEGF, the densities of macrophages, fibroblasts, capillary tips and sprouts, and the density and velocity of the ECM. Simulations of the model demonstrate how ischemic conditions may limit macrophage recruitment to the wound-site and impair wound closure. The results are in general agreement with experimental findings. PMID- 19805374 TI - From the Cover: CD39 deletion exacerbates experimental murine colitis and human polymorphisms increase susceptibility to inflammatory bowel disease. AB - CD39/ENTPD1 hydrolyzes proinflammatory nucleotides to generate adenosine. As purinergic mediators have been implicated in intestinal inflammation, we hypothesized that CD39 might protect against inflammatory bowel disease. We studied these possibilities in a mouse model of colitis using mice with global CD39 deletion. We then tested whether human genetic polymorphisms in the CD39 gene might influence susceptibility to Crohn's disease. We induced colitis in mice using Dextran Sodium Sulfate (DSS). Readouts included disease activity scores, histological evidence of injury, and markers of inflammatory activity. We used HapMap cell lines to find SNPs that tag for CD39 expression, and then compared the frequency of subjects with high vs. low CD39-expression genotypes in a case-control cohort for Crohn's disease. Mice null for CD39 were highly susceptible to DSS injury, with heterozygote mice showing an intermediate phenotype compared to wild type (WT). We identified a common SNP that tags CD39 mRNA expression levels in man. The SNP tagging low levels of CD39 expression was associated with increased susceptibility to Crohn's disease in a case-control cohort comprised of 1,748 Crohn's patients and 2,936 controls (P = 0.005-0.0006). Our data indicate that CD39 deficiency exacerbates murine colitis and suggest that CD39 polymorphisms are associated with inflammatory bowel disease in humans. PMID- 19805375 TI - Regulation of proto-oncogene transcription, cell proliferation, and tumorigenesis in mice by PSF protein and a VL30 noncoding RNA. AB - We describe the role of PSF protein and VL30-1 RNA, a mouse retroelement noncoding RNA, in the reversible regulation of proto-oncogene transcription, cell proliferation, and tumorigenesis in mice. The experiments involved increasing expression of PSF or VL30-1 RNA in NIH/3T3 fibroblast cells and B16F10 melanoma cells by transfecting the respective coding genes under control of a strong promoter or decreasing expression by transfecting a shRNA construct that causes degradation of PSF mRNA or VL30-1 RNA. The results are as follows: (i) PSF binds to the proto-oncogene Rab23, repressing transcription, and VL30-1 RNA binds and releases PSF from Rab23, activating transcription; (ii) increasing expression of PSF or decreasing expression of VL30-1 RNA suppresses cell proliferation in culture and tumorigenesis in mice; and (iii) decreasing expression of PSF or increasing expression of VL30-1 RNA promotes cell proliferation in culture and tumorigenesis in mice. These results indicate that PSF is a major tumor suppressor protein and VL30-1 RNA is a major tumor-promoter RNA in mice. Although VL30-1 RNA can integrate into the cell genome, tumor promotion by VL30-1 RNA involves a trans effect rather than a cis effect on gene transcription. Expression of VL30-1 RNA is 5- to 8-fold higher in mouse tumor lines than in mouse fibroblast or myoblast lines, whereas expression of PSF mRNA does not decrease in the tumor lines, suggesting that tumorigenesis is driven by an increase of VL30-1 RNA rather than a decrease of PSF. A similar regulatory mechanism functions in human cells, except that human PSF-binding RNAs replace VL30-1 RNA, which is not encoded in the human genome. We propose that PSF protein and PSF-binding RNAs have a central role in the reversible regulation of mammalian cell proliferation and tumorigenesis and that increasing PSF expression or decreasing PSF-binding RNA expression in tumor cells is a potential therapeutic strategy for cancer. PMID- 19805376 TI - Identification of human zonulin, a physiological modulator of tight junctions, as prehaptoglobin-2. AB - Increased intestinal permeability (IP) has emerged recently as a common underlying mechanism in the pathogenesis of allergic, inflammatory, and autoimmune diseases. The characterization of zonulin, the only physiological mediator known to regulate IP reversibly, has remained elusive. Through proteomic analysis of human sera, we have now identified human zonulin as the precursor for haptoglobin-2 (pre-HP2). Although mature HP is known to scavenge free hemoglobin (Hb) to inhibit its oxidative activity, no function has ever been ascribed to its uncleaved precursor form. We found that the single-chain zonulin contains an EGF like motif that leads to transactivation of EGF receptor (EGFR) via proteinase activated receptor 2 (PAR(2)) activation. Activation of these 2 receptors was coupled to increased IP. The siRNA-induced silencing of PAR(2) or the use of PAR(2)(-/-) mice prevented loss of barrier integrity. Proteolytic cleavage of zonulin into its alpha(2)- and beta-subunits neutralized its ability to both activate EGFR and increase IP. Quantitative gene expression revealed that zonulin is overexpressed in the intestinal mucosa of subjects with celiac disease. To our knowledge, this is the initial example of a molecule that exerts a biological activity in its precursor form that is distinct from the function of its mature form. Our results therefore characterize zonulin as a previously undescribed ligand that engages a key signalosome involved in the pathogenesis of human immune-mediated diseases that can be targeted for therapeutic interventions. PMID- 19805377 TI - HIF-2alpha maintains an undifferentiated state in neural crest-like human neuroblastoma tumor-initiating cells. AB - High hypoxia-inducible factor-2alpha (HIF-2alpha) protein levels predict poor outcome in neuroblastoma, and hypoxia dedifferentiates cultured neuroblastoma cells toward a neural crest-like phenotype. Here, we identify HIF-2alpha as a marker of normoxic neural crest-like neuroblastoma tumor-initiating/stem cells (TICs) isolated from patient bone marrows. Knockdown of HIF-2alpha reduced VEGF expression and induced partial sympathetic neuronal differentiation when these TICs were grown in vitro under stem cell-promoting conditions. Xenograft tumors of HIF-2alpha-silenced cells were widely necrotic, poorly vascularized, and resembled the bulk of tumor cells in clinical neuroblastomas by expressing additional sympathetic neuronal markers, whereas control tumors were immature, well-vascularized, and stroma-rich. Thus, HIF-2alpha maintains an undifferentiated state of neuroblastoma TICs. Because low differentiation is associated with poor outcome and angiogenesis is crucial for tumor growth, HIF 2alpha is an attractive target for neuroblastoma therapy. PMID- 19805378 TI - Reduction of phosphoinositide-phospholipase C beta1 methylation predicts the responsiveness to azacitidine in high-risk MDS. AB - Lipid signaling pathways are involved in cell growth, differentiation, and apoptosis, and could have a role in the progression of myelodysplastic syndromes (MDS) into acute myeloid leukemia (AML). Indeed, recent studies showed that phosphoinositide-phospholipase (PI-PL)Cbeta1 mono-allelic deletion correlates with a higher risk of AML evolution. Also, a single patient treated with azacitidine, a DNA methyltransferase inhibitor currently used in MDS, displayed a direct correlation between PI-PLCbeta1 gene expression and drug responsiveness. Consequently, we hypothesized that PI-PLCbeta1 could be a target for demethylating therapy. First, we analyzed the structure of PI-PLCbeta1 gene promoter, then quantified the degree of PI-PLCbeta1 promoter methylation and gene expression in MDS patients at baseline and during azacitidine administration. Indeed, PI-PLCbeta1 mRNA increased in responder patients, along with a reduction of PI-PLCbeta1 promoter methylation. Also, the molecular response correlated to and anticipated the clinical outcome, thus suggesting that PI-PLCbeta1 gene reactivation could predict azacitidine responsiveness. Our results demonstrate not only that PI-PLCbeta1 promoter is hypermethylated in high-risk MDS patients, but also that the amount of PI-PLCbeta1 mRNA could predict the clinical response to azacitidine, therefore indicating a promising new therapeutic approach. PMID- 19805379 TI - Chromatin protein HMGB2 regulates articular cartilage surface maintenance via beta-catenin pathway. AB - The superficial zone (SZ) of articular cartilage is critical in maintaining tissue function and homeostasis and represents the site of the earliest changes in osteoarthritis. Mechanisms that regulate the unique phenotype of SZ chondrocytes and maintain SZ integrity are unknown. We recently demonstrated that expression of the chromatin protein high mobility group box (HMGB) protein 2 is restricted to the SZ in articular cartilage suggesting a transcriptional regulation involving HMGB2 in SZ. Here, we show that an interaction between HMGB2 and the Wnt/beta-catenin pathway regulates the maintenance of the SZ. We found that the Wnt/beta-catenin pathway is active specifically in the SZ in normal mouse knee joints and colocalizes with HMGB2. Both Wnt signaling and HMGB2 expression decrease with aging in mouse joints. Our molecular studies show that HMGB2 enhances the binding of Lef-1 to its target sequence and potentiates transcriptional activation of the Lef-1-beta-catenin complex. The HMG domain within HMGB2 is crucial for interaction with Lef-1, suggesting that both HMGB2 and HMGB1 may be involved in this function. Furthermore, conditional deletion of beta-catenin in cultured mouse chondrocytes induced apoptosis. These findings define a pathway where protein interactions of HMGB2 and Lef-1 enhance Wnt signaling and promote SZ chondrocyte survival. Loss of the HMGB2-Wnt signaling interaction is a new mechanism in aging-related cartilage pathology. PMID- 19805381 TI - A state-mutating genetic algorithm to design ion-channel models. AB - Realistic computational models of single neurons require component ion channels that reproduce experimental findings. Here, a topology-mutating genetic algorithm that searches for the best state diagram and transition-rate parameters to model macroscopic ion-channel behavior is described. Important features of the algorithm include a topology-altering strategy, automatic satisfaction of equilibrium constraints (microscopic reversibility), and multiple-protocol fitting using sequential goal programming rather than explicit weighting. Application of this genetic algorithm to design a sodium-channel model exhibiting both fast and prolonged inactivation yields a six-state model that produces realistic activity-dependent attenuation of action-potential backpropagation in current-clamp simulations of a CA1 pyramidal neuron. PMID- 19805380 TI - Human papillomavirus 16 E7 inactivator of retinoblastoma family proteins complements human cytomegalovirus lacking UL97 protein kinase. AB - Several different families of DNA viruses encode proteins that inactivate the cellular retinoblastoma tumor suppressor protein (pRb), which normally functions to bind E2F transcription factors and restrict expression of genes necessary for cellular processes including DNA replication. Human cytomegalovirus (HCMV) UL97, a protein kinase functionally orthologous to cellular cyclin-dependent kinases, phosphorylates pRb on inactivating residues during HCMV infection. To assess if such phosphorylation is biologically relevant, we tested whether the human papillomavirus type 16 E7 protein, which inactivates pRb family proteins by direct binding and destabilization, could substitute for UL97 during HCMV infection. In the absence of UL97, expression of wild-type E7 protein, but not a mutant E7 unable to bind pRb family proteins, restored E2F-responsive cellular gene expression, late viral gene expression, and viral DNA synthesis to levels normally observed during wild-type virus infection of quiescent cells. UL97-null mutants exhibited more pronounced defects in virus production and DNA synthesis in quiescent cells as compared to serum-fed, cycling cells. E7 expression substantially enhanced infectious virus production in quiescent cells, but did not complement the defects observed during UL97-null virus infection of cycling cells. Thus, a primary role of UL97 is to inactivate pRb family proteins during infection of quiescent cells, and this inactivation likely abets virus replication by induction of cellular E2F-responsive genes. Our findings have implications for human cytomegalovirus disease and for drugs that target UL97. PMID- 19805382 TI - Neural responses to sanction threats in two-party economic exchange. AB - Sanctions are used ubiquitously to enforce obedience to social norms. However, recent field studies and laboratory experiments have demonstrated that cooperation is sometimes reduced when incentives meant to promote prosocial decisions are added to the environment. Although various explanations for this effect have been suggested, the neural foundations of the effect have not been fully explored. Using a modified trust game, we found that trustees reciprocate relatively less when facing sanction threats, and that the presence of sanctions significantly reduces trustee's brain activities involved in social reward valuation [in the ventromedial prefrontal cortex (VMPFC), lateral orbitofrontal cortex, and amygdala] while it simultaneously increases brain activities in the parietal cortex, which has been implicated in rational decision making. Moreover, we found that neural activity in a trustee's VMPFC area predicts her future level of cooperation under both sanction and no-sanction conditions, and that this predictive activity can be dynamically modulated by the presence of a sanction threat. PMID- 19805383 TI - Segregating the significant from the mundane on a moment-to-moment basis via direct and indirect amygdala contributions. AB - If the amygdala is involved in shaping perceptual experience when affectively significant visual items are encountered, responses in this structure should be correlated with both visual cortex responses and behavioral reports. Here, we investigated how affective significance shapes visual perception during an attentional blink paradigm combined with aversive conditioning. Behaviorally, following aversive learning, affectively significant scenes (CS(+)) were better detected than neutral (CS(-)) ones. In terms of mean brain responses, both amygdala and visual cortical responses were stronger during CS(+) relative to CS( ) trials. Increased brain responses in these regions were associated with improved behavioral performance across participants and followed a mediation-like pattern. Importantly, the mediation pattern was observed in a trial-by-trial analysis, revealing that the specific pattern of trial-by-trial variability in brain responses was closely related to single-trial behavioral performance. Furthermore, the influence of the amygdala on visual cortical responses was consistent with a mediation, although partial, via frontal brain regions. Our results thus suggest that affective significance potentially determines the fate of a visual item during competitive interactions by enhancing sensory processing through both direct and indirect paths. In so doing, the amygdala helps separate the significant from the mundane. PMID- 19805384 TI - C-type natriuretic peptide (CNP) is a bifurcation factor for sensory neurons. AB - Neuronal circuits are shaped during development by the coordinated action of guidance factors and signals that regulate axonal branching. Unlike guidance cues, the molecules and signaling cascades that underlie axonal branching remain to be resolved. Here we show that the secreted molecule C-type natriuretic peptide (CNP) induces a cGMP signaling cascade via its receptor particulate guanylyl cyclase Npr2 which is essential for sensory axon bifurcation at the dorsal root entry zone (DREZ) of the spinal cord. In contrast, another form of sensory axon branching-collateral formation-is not affected by this pathway. We also demonstrate that cGMP signaling via the nitric oxide-stimulated soluble guanylyl cyclase system (NO-GC) is dispensable for sensory axon branching. Functionally, the bifurcation error in CNP mutant mice is maintained at mature stages and results in a reduced input on secondary neurons as detected by patch clamp recordings. PMID- 19805385 TI - Modulation of neuronal responses during covert search for visual feature conjunctions. AB - While searching for an object in a visual scene, an observer's attentional focus and eye movements are often guided by information about object features and spatial locations. Both spatial and feature-specific attention are known to modulate neuronal responses in visual cortex, but little is known of the dynamics and interplay of these mechanisms as visual search progresses. To address this issue, we recorded from directionally selective cells in visual area MT of monkeys trained to covertly search for targets defined by a unique conjunction of color and motion features and to signal target detection with an eye movement to the putative target. Two patterns of response modulation were observed. One pattern consisted of enhanced responses to targets presented in the receptive field (RF). These modulations occurred at the end-stage of search and were more potent during correct target identification than during erroneous saccades to a distractor in RF, thus suggesting that this modulation is not a mere presaccadic enhancement. A second pattern of modulation was observed when RF stimuli were nontargets that shared a feature with the target. The latter effect was observed during early stages of search and is consistent with a global feature-specific mechanism. This effect often terminated before target identification, thus suggesting that it interacts with spatial attention. This modulation was exhibited not only for motion but also for color cue, although MT neurons are known to be insensitive to color. Such cue-invariant attentional effects may contribute to a feature binding mechanism acting across visual dimensions. PMID- 19805386 TI - Loss of dendrite stabilization by the Abl-related gene (Arg) kinase regulates behavioral flexibility and sensitivity to cocaine. AB - Adolescence is characterized by increased vulnerability to developing neuropsychiatric disorders and involves a period of prefrontal cortical dendritic refinement and synaptic pruning that culminates in cytoskeletal stabilization in adulthood. The Abl-related gene (Arg) acts through p190RhoGAP to inhibit the RhoA GTPase and stabilize cortical dendritic arbors beginning in adolescence. Cortical axons, dendrites, and synapses develop normally in Arg-deficient (arg(-/-)) mice, but adult dendrites destabilize and regress; thus, arg(-/-) mice present a model of adolescent-onset dendritic simplification. We show that arg(-/-) mice are impaired in a reversal task and that deficits are grossly exacerbated by low-dose cocaine administration. Although ventral prefrontal dopamine D2 receptor levels predict "perseverative" error counts in wild-type mice, no such relationship is found in arg(-/-) mice. Moreover, arg(-/-) mice are insensitive to the disruptive effects of the D2/D3 antagonist haloperidol in reversal but show normal sensitivity to its locomotor-depressant actions. Arg deficiency and orbitofrontal cortical Arg inhibition via STI-571 infusion also enhance the psychomotor stimulant actions of cocaine. These findings provide evidence that stabilization of dendritic structure beginning in adolescence is critical for the development of adaptive and flexible behavior after cocaine exposure. PMID- 19805387 TI - Experience-dependent maturation of the glomerular microcircuit. AB - Spontaneous and patterned activity, largely attributed to chemical transmission, shape the development of virtually all neural circuits. However, electrical transmission also has an important role in coordinated activity in the brain. In the olfactory bulb, gap junctions between apical dendrites of mitral cells increase excitability and synchronize firing within each glomerulus. We report here that the development of the glomerular microcircuit requires both sensory experience and connexin (Cx)36-mediated gap junctions. Coupling coefficients, which measure electrical coupling between mitral cell dendrites, were high in young mice, but decreased after postnatal day (P)10 because of a maturational increase in membrane conductance. Sensory deprivation, induced by unilateral naris occlusion at birth, slowed the morphological development of mitral cells and arrested the maturational changes in membrane conductance and coupling coefficients. As the coupling coefficients decreased in normal mice, a glutamate mediated excitatory postsynaptic current (EPSC) between mitral cells emerged by P30. Although mitral-mitral EPSCs were generally unidirectional, they were not present in young adult Cx36(-/-) mice, suggesting that gap junctions are required for the development and/or function of the mature circuit. The experience dependent transition from electrical transmission to combined chemical and electrical transmission provides a previously unappreciated mechanism that may tune the response properties of the glomerular microcircuit. PMID- 19805388 TI - DeltaNp73 regulates neuronal survival in vivo. AB - Apoptosis occurs widely during brain development, and p73 transcription factors are thought to play essential roles in this process. The p73 transcription factors are present in two forms, the full length TAp73 and the N-terminally truncated DeltaNp73. In cultured sympathetic neurons, overexpression of DeltaNp73 inhibits apoptosis induced by nerve growth factor withdrawal or p53 overexpression. To probe the function of DeltaNp73 in vivo, we generated a null allele and inserted sequences encoding the recombinase Cre and green fluorescent protein (EGFP). We show that DeltaNp73 is heavily expressed in the thalamic eminence (TE) that contributes neurons to ventral forebrain, in vomeronasal neurons, Cajal-Retzius cells (CRc), and choroid plexuses. In DeltaNp73(-/-) mice, cells in preoptic areas, vomeronasal neurons, GnRH-positive cells, and CRc were severely reduced in number, and choroid plexuses were atrophic. This phenotype was enhanced when DeltaNp73-positive cells were ablated by diphtheria toxin expression. However, ablation of cells that express DeltaNp73 and Wnt3a did neither remove all CRc, nor did they abolish Reelin secretion or generate a reeler-like cortical phenotype. Our data emphasize the role of DeltaNp73 in neuronal survival in vivo and in choroid plexus development, the importance of the TE as a source of neurons in ventral forebrain, and the multiple origins of CRc, with redundant production of Reelin. PMID- 19805389 TI - Reversal of long-term dendritic spine alterations in Alzheimer disease models. AB - Synapse loss is strongly correlated with cognitive impairment in Alzheimer's disease (AD). We have previously reported the loss of dendritic spines and the presence of dystrophic neurites in both the hippocampi of transgenic mice overexpressing amyloid precursor protein (APP) and in the human brain affected with AD. In the studies reported here we have asked whether the acute alterations in dendritic spines induced by Abeta, as well as the chronic loss of spine density seen in hAPP transgenic mice, are reversible by treatments that restore the cAMP/PKA/CREB signaling pathway or proteasome function to control levels. The results show that both rolipram and TAT-HA-Uch-L1 restore spine density to near control conditions, even in elderly mice. The results suggest that changes in dendritic structure and function that occur after Abeta elevation are reversible even after long periods of time, and that one could envision therapeutic approaches to AD based on this restoration that could work independently of therapies aimed at lowering Abeta levels in the brain. PMID- 19805390 TI - REVEILLE1, a Myb-like transcription factor, integrates the circadian clock and auxin pathways. AB - The circadian clock modulates expression of a large fraction of the Arabidopsis genome and affects many aspects of plant growth and development. We have discovered one way in which the circadian system regulates hormone signaling, identifying a node that links the clock and auxin networks. Auxin plays key roles in development and responses to environmental cues, in part through regulation of plant growth. We have characterized REVEILLE1 (RVE1), a Myb-like, clock-regulated transcription factor that is homologous to the central clock genes CIRCADIAN CLOCK ASSOCIATED 1 (CCA1) and LATE ELONGATED HYPOCOTYL (LHY). Despite this homology, inactivation of RVE1 does not affect circadian rhythmicity but instead causes a growth phenotype, indicating this factor is a clock output affecting plant development. CCA1 regulates growth via the bHLH transcription factors PHYTOCHROME INTERACTING FACTOR4 (PIF4) and PIF5, but RVE1 acts independently of these genes. RVE1 instead controls auxin levels, promoting free auxin production during the day but having no effect during the night. RVE1 positively regulates the expression of the auxin biosynthetic gene YUCCA8 (YUC8), providing a mechanism for its growth-promoting effects. RVE1 is therefore a node that connects two important signaling networks that coordinate plant growth with rhythmic changes in the environment. PMID- 19805391 TI - Evaluation of five decontamination methods for filtering facepiece respirators. AB - Concerns have been raised regarding the availability of National Institute for Occupational Safety and Health (NIOSH)-certified N95 filtering facepiece respirators (FFRs) during an influenza pandemic. One possible strategy to mitigate a respirator shortage is to reuse FFRs following a biological decontamination process to render infectious material on the FFR inactive. However, little data exist on the effects of decontamination methods on respirator integrity and performance. This study evaluated five decontamination methods [ultraviolet germicidal irradiation (UVGI), ethylene oxide, vaporized hydrogen peroxide (VHP), microwave oven irradiation, and bleach] using nine models of NIOSH-certified respirators (three models each of N95 FFRs, surgical N95 respirators, and P100 FFRs) to determine which methods should be considered for future research studies. Following treatment by each decontamination method, the FFRs were evaluated for changes in physical appearance, odor, and laboratory performance (filter aerosol penetration and filter airflow resistance). Additional experiments (dry heat laboratory oven exposures, off-gassing, and FFR hydrophobicity) were subsequently conducted to better understand material properties and possible health risks to the respirator user following decontamination. However, this study did not assess the efficiency of the decontamination methods to inactivate viable microorganisms. Microwave oven irradiation melted samples from two FFR models. The remainder of the FFR samples that had been decontaminated had expected levels of filter aerosol penetration and filter airflow resistance. The scent of bleach remained noticeable following overnight drying and low levels of chlorine gas were found to off-gas from bleach decontaminated FFRs when rehydrated with deionized water. UVGI, ethylene oxide (EtO), and VHP were found to be the most promising decontamination methods; however, concerns remain about the throughput capabilities for EtO and VHP. Further research is needed before any specific decontamination methods can be recommended. PMID- 19805393 TI - Effects of headspace and oxygen level on off-gas emissions from wood pellets in storage. AB - Few papers have been published in the open literature on the emissions from biomass fuels, including wood pellets, during the storage and transportation and their potential health impacts. The purpose of this study is to provide data on the concentrations, emission factors, and emission rate factors of CO(2), CO, and CH(4) from wood pellets stored with different headspace to container volume ratios with different initial oxygen levels, in order to develop methods to reduce the toxic off-gas emissions and accumulation in storage spaces. Metal containers (45 l, 305 mm diameter by 610 mm long) were used to study the effect of headspace and oxygen levels on the off-gas emissions from wood pellets. Concentrations of CO(2), CO, and CH(4) in the headspace were measured using a gas chromatograph as a function of storage time. The results showed that the ratio of the headspace ratios and initial oxygen levels in the storage space significantly affected the off-gas emissions from wood pellets stored in a sealed container. Higher peak emission factors and higher emission rates are associated with higher headspace ratios. Lower emissions of CO(2) and CO were generated at room temperature under lower oxygen levels, whereas CH(4) emission is insensitive to the oxygen level. Replacing oxygen with inert gases in the storage space is thus a potentially effective method to reduce the biomass degradation and toxic off gas emissions. The proper ventilation of the storage space can also be used to maintain a high oxygen level and low concentrations of toxic off-gassing compounds in the storage space, which is especially useful during the loading and unloading operations to control the hazards associated with the storage and transportation of wood pellets. PMID- 19805392 TI - Quantitative plasma biomarker analysis in HDI exposure assessment. AB - Quantification of amines in biological samples is important for evaluating occupational exposure to diisocyanates. In this study, we describe the quantification of 1,6-hexamethylene diamine (HDA) levels in hydrolyzed plasma of 46 spray painters applying 1,6-hexamethylene diisocyanate (HDI)-containing paint in vehicle repair shops collected during repeated visits to their workplace and their relationship with dermal and inhalation exposure to HDI monomer. HDA was detected in 76% of plasma samples, as heptafluorobutyryl derivatives, and the range of HDA concentrations was < or =0.02-0.92 microg l(-1). After log transformation of the data, the correlation between plasma HDA levels and HDI inhalation exposure measured on the same workday was low (N = 108, r = 0.22, P = 0.026) compared with the correlation between plasma HDA levels and inhalation exposure occurring approximately 20 to 60 days before blood collection (N = 29, r = 0.57, P = 0.0014). The correlation between plasma HDA levels and HDI dermal exposure measured on the same workday, although statistically significant, was low (N = 108, r = 0.22, P = 0.040) while the correlation between HDA and dermal exposure occurring approximately 20 to 60 days before blood collection was slightly improved (N = 29, r = 0.36, P = 0.053). We evaluated various workplace factors and controls (i.e. location, personal protective equipment use and paint booth type) as modifiers of plasma HDA levels. Workers using a downdraft ventilated booth had significantly lower plasma HDA levels relative to semi downdraft and crossdraft booth types (P = 0.0108); this trend was comparable to HDI inhalation and dermal exposure levels stratified by booth type. These findings indicate that HDA concentration in hydrolyzed plasma may be used as a biomarker of cumulative inhalation and dermal exposure to HDI and for investigating the effectiveness of exposure controls in the workplace. PMID- 19805394 TI - A comparative assessment of the forces and moments generated with various maxillary incisor intrusion biomechanics. AB - The aim of this study was to comparatively evaluate the intrusive forces and buccolingual torquing moments generated during anterior maxillary intrusion using different maxillary incisor intrusion mechanics. Five wire specimens were used for each of the following intrusive arches: blue Elgiloy utility arch 0.016 x 0.016 inch, TMA utility arch 0.017 x 0.025 inch, Burstone TMA intrusion arch 0.017 x 0.025 inch, and reverse curve of Spee NiTi 0.016 x 0.022 inch. The wires were inserted on bracketed dental arches constructed on maxillary Frasaco models, segmented mesially to the maxillary canines. Simulated intrusion from 0.0 to 3 mm was performed using the orthodontic measurement and simulation system (OMSS), and forces and moments were recorded in the sagittal plane at 0.1 mm vertical displacement increments. All measurements were repeated five times for each specimen and values recorded at 1.5 mm for all wires were used for statistical evaluations. The results were analysed with one-way analysis of variance with forces and moments serving as the dependent variables and wire type as the independent variable. Post hoc multiple comparisons were performed using the Tukey test (0.05 error rate). Comparison of the two major intrusion techniques for the maxillary anterior teeth, segmented and bioprogressive, revealed that the Burstone TMA 0.017 x 0.025 inch intrusion arch exerted the lowest force on the incisors (0.99 N), followed by the TMA utility 0.017 x 0.025 inch (1.33 N) and the blue Elgiloy 0.016 x 0.016 inch utility (1.43 N). The highest force was recorded for the reverse curve of Spee NiTi and exceeded the value of 9 N. The lowest buccolingual moments were recorded with the Burstone intrusion arch (2.47 Nmm), whereas the highest was registered for the utility arch constructed with a 0.017 x 0.025 inch TMA wire (7.31 Nmm). PMID- 19805395 TI - The amylose extender mutant of maize conditions novel protein-protein interactions between starch biosynthetic enzymes in amyloplasts. AB - The amylose extender (ae(-)) mutant of maize lacks starch branching enzyme IIb (SBEIIb) activity, resulting in amylopectin with reduced branch point frequency, and longer glucan chains. Recent studies indicate isozymes of soluble starch synthases form high molecular weight complexes with SBEII isoforms. This study investigated the effect of the loss of SBEIIb activity on interactions between starch biosynthetic enzymes in maize endosperm amyloplasts. Results show distinct patterns of protein-protein interactions in amyloplasts of ae(-) mutants compared with the wild type, suggesting functional complementation for loss of SBEIIb by SBEI, SBEIIa, and SP. Coimmunoprecipitation experiments and affinity chromatography using recombinant proteins showed that, in amyloplasts from normal endosperm, protein-protein interactions involving starch synthase I (SSI), SSIIa, and SBEIIb could be detected. By contrast, in ae(-) amyloplasts, SSI and SSIIa interacted with SBEI, SBEIIa, and SP. All interactions in the wild-type were strongly enhanced by ATP, and broken by alkaline phosphatase, indicating a role for protein phosphorylation in their assembly. Whilst ATP and alkaline phosphatase had no effect on the stability of the protein complexes from ae(-) endosperm, radiolabelling experiments showed SP and SBEI were both phosphorylated within the mutant protein complex. It is proposed that, during amylopectin biosynthesis, SSI and SSIIa form the core of a phosphorylation-dependent glucan synthesizing protein complex which, in normal endosperm, recruits SBEIIb, but when SBEIIb is absent (ae(-)), recruits SBEI, SBEIIa, and SP. Differences in stromal protein complexes are mirrored in the complement of the starch synthesizing enzymes detected in the starch granules of each genotype, reinforcing the hypothesis that the complexes play a functional role in starch biosynthesis. PMID- 19805396 TI - High job control enhances vagal recovery in media work. AB - BACKGROUND: Job strain has been linked to increased risk of cardiovascular diseases. In modern media work, time pressures, rapidly changing situations, computer work and irregular working hours are common. Heart rate variability (HRV) has been widely used to monitor sympathovagal balance. Autonomic imbalance may play an additive role in the development of cardiovascular diseases. AIMS: To study the effects of work demands and job control on the autonomic nervous system recovery among the media personnel. METHODS: From the cross-sectional postal survey of the employees in Finnish Broadcasting Company (n = 874), three age cohorts (n = 132) were randomly selected for an analysis of HRV in 24 h electrocardiography recordings. RESULTS: In the middle-aged group, those who experienced high job control had significantly better vagal recovery than those with low or moderate control (P < 0.01). Among young and ageing employees, job control did not associate with autonomic recovery. CONCLUSIONS: High job control over work rather than low demands seemed to enhance autonomic recovery in middle aged media workers. This was independent of poor health habits such as smoking, physical inactivity or alcohol consumption. PMID- 19805397 TI - Suicides among seafarers in UK merchant shipping, 1919-2005. AB - BACKGROUND: Little has been reported on suicides among seafarers and how they have changed over time. AIMS: To establish the causes, rates and trends in suicides at work among seafarers in UK merchant shipping from 1919 to 2005 and to compare suicide rates with the general UK population and with seafarers employed in non-UK shipping. METHODS: Examination of seafarers' death inquiry files, death registers and death returns (for a total population of 11.90 million seafarer years); literature reviews and national suicide statistics. RESULTS: The suicide rate (for suicides at work and unexplained disappearances at sea) in UK shipping fell from 40-50 per 100,000 in the 1920s to <10 per 100 000 in recent years, with an interim peak during the 1960s. Suicide rates were higher for ratings (all ranks below officers) than for officers, for Lascars (Asian seafarers) than for British seafarers and for older than for younger seafarers and were typically lower than those in Asian and Scandinavian merchant fleets. The suicide rate (for suicides at work) among seafarers was substantially higher than the overall suicide rate in the general British population from 1919 to the 1970s, but following reductions in suicide mortality among seafarers, it has become more comparable since. CONCLUSIONS: Although merchant seafaring was previously a high risk occupation for suicides at work, there has been a sharp fall in the suicide rate in the past 40 years. Likely reasons for this include reductions over time in long intercontinental voyages and changes over time in seafarers' lifestyles. PMID- 19805398 TI - Multidimensional intervention and sickness absence in assistant nursing students. AB - BACKGROUND: When handling patients, nursing assistant (NA) students and nurse students are frequently exposed to risk factors for low back pain (LBP) including sudden loads and twisting and bending of the spine. Furthermore, LBP is a major cause of sickness absence. AIMS: To ascertain if a multidimensional prevention programme combining physical training, patient transfer technique and stress management prevents sickness absence and LBP in NA students. METHODS: The study was a 14-month cluster randomized controlled study. The participants were NA students from 37 randomly selected classes located at two schools of health and social care in Copenhagen, Denmark. The participants completed a comprehensive questionnaire regarding sickness absence, LBP and psychosocial factors on commencement and after completion of the study. RESULTS: Of 766 female NA students, 668 (87%) completed the baseline questionnaire. Sickness absence during the study period increased in both groups but the increase was significantly lower in the intervention group than the control group, mean (standard deviation) number of days 12 (20) versus 18 (34), P < 0.05. The intervention group reported no change in the mean level of general health perception, energy/fatigue or psychological well-being at follow-up, while the control group reported a decline on those scales. There were no significant differences in the prevalence of LBP at follow-up between the intervention and control group. CONCLUSIONS: Compared to the control group, the intervention group had significantly less sickness absence. The intervention had no preventive effect on LBP prevalence. PMID- 19805399 TI - Monocytes in heart failure: relationship to a deteriorating immune overreaction or a desperate attempt for tissue repair? AB - Monocytes play an important role in immune defence, inflammation, and tissue remodelling. Nevertheless, the role of monocytes in cardiovascular disease is obscure. Indeed, monocytes infiltrate dysfunctional tissue and augment tissue damage and are actively involved in tissue regeneration and healing. In support of the latter, recent studies have provided data on the functional and structural plasticity of monocytes. Monocytes are also actively involved in processes associated with tissue regeneration such as angiogenesis and vasculogenesis, either by producing pro-angiogenic factors or even by evolving to structural components of the vascular wall. This review article provides an overview on whether monocytes represent deteriorating immune overreaction in heart failure (HF), or a desperate attempt for tissue repair or physiological compensation in the failing heart. Perhaps, it is time to reconsider our attitude towards monocytes and consider more 'monocyte activation' rather than 'monocyte suppression' as a potential therapeutic target in HF. PMID- 19805400 TI - Dual effects of superovulation: loss of maternal and paternal imprinted methylation in a dose-dependent manner. AB - Superovulation or ovarian stimulation is currently an indispensable assisted reproductive technology (ART) for human subfertility/infertility treatment. Recently, increased frequencies of imprinting disorders have been correlated with ARTs. Significantly, for Angelman and Beckwith-Wiedemann Syndromes, patients have been identified where ovarian stimulation was the only procedure used by the couple undergoing ART. In many cases, increased risk of genomic imprinting disorders has been attributed to superovulation in combination with inherent subfertility. To distinguish between these contributing factors, carefully controlled experiments are required on spontaneously ovulated, in vivo-fertilized oocytes and their induced-ovulated counterparts, thereby minimizing effects of in vitro manipulations. To this end, effects of superovulation on genomic imprinting were evaluated in a mouse model, where subfertility is not a confounding issue. This work represents the first comprehensive examination of the overall effects of superovulation on imprinted DNA methylation for four imprinted genes in individual blastocyst stage embryos. We demonstrate that superovulation perturbed genomic imprinting of both maternally and paternally expressed genes; loss of Snrpn, Peg3 and Kcnq1ot1 and gain of H19 imprinted methylation were observed. This perturbation was dose-dependent, with aberrant imprinted methylation more frequent at the high hormone dosage. Superovulation is thought to primarily affect oocyte development; thus, effects were expected to be limited to maternal alleles. Our study revealed that maternal as well as paternal H19 methylation was perturbed by superovulation. We postulate that superovulation has dual effects during oogenesis, disrupting acquisition of imprints in growing oocytes, as well as maternal-effect gene products subsequently required for imprint maintenance during pre-implantation development. PMID- 19805402 TI - Bandaru s. Reddy: in memoriam (1930-2009). PMID- 19805403 TI - Adaptive significance and ontogenetic variability of the waxy zone in Nepenthes rafflesiana. AB - BACKGROUND AND AIMS: The slippery waxy zone in the upper part of pitchers has long been considered the key trapping structure of the Nepenthes carnivorous plants; however, the presence of wax is reported to be variable within and between species of this species-rich genus. This study raises the question of the adaptive significance of the waxy zone and investigates the basis for an ontogenetic cause of its variability and correlation with pitcher shape. METHODS: In Brunei (Borneo) the expression of the waxy zone throughout plant ontogeny was studied in two taxa of the Nepenthes rafflesiana complex, typica and elongata, which differ in pitcher shape and size. We also tested the adaptive significance of this zone by comparing the trapping efficiency and the number of prey captured of wax-bearing and wax-lacking plants. KEY RESULTS: In elongata, the waxy zone is always well expanded and the elongated pitchers change little in form during plant development. Wax efficiently traps experimental ants but the number of captured prey in pitchers is low. In contrast, in typica, the waxy zone is reduced in successively produced pitchers until it is lost at the end of the plant's juvenile stage. The form of pitchers thus changes continuously throughout plant ontogeny, from elongated to ovoid. In typica, the number of captured prey is greater, but the role of wax in trapping is minor compared with that of the digestive liquid, and waxy plants do not show a higher insect retention and prey abundance as compared with non-waxy plants. CONCLUSIONS: The waxy zone is not always a key trapping structure in Nepenthes and can be lost when supplanted by more efficient features. This study points out how pitcher structure is submitted to selection, and that evolutionary changes in developmental mechanisms could play a role in the morphological diversity of Nepenthes. PMID- 19805401 TI - Racial discrimination and health among Asian Americans: evidence, assessment, and directions for future research. AB - Research shows that racial discrimination is related to illness among diverse racial and ethnic populations. Studies of racial discrimination and health among Asian Americans, however, remain underdeveloped. In this paper, the authors review evidence on racial discrimination and health among Asian Americans, identify gaps in the literature, and provide suggestions for future research. They identified 62 empirical articles assessing the relation between discrimination and health among Asian Americans. The majority of articles focused on mental health problems, followed by physical and behavioral problems. Most studies find that discrimination was associated with poorer health, although the most consistent findings were for mental health problems. This review suggests that future studies should continue to investigate the following: 1) the measurement of discrimination among Asian Americans, whose experiences may be qualitatively different from those of other racial minority groups; 2) the heterogeneity among Asian Americans, including those factors that are particularly salient in this population, such as ethnic ancestry and immigration history; and 3) the health implications of discrimination at multiple ecologic levels, ranging from the individual level to the structural level. PMID- 19805404 TI - Plant morphometric traits and climate gradients in northern China: a meta analysis using quadrat and flora data. AB - BACKGROUND AND AIMS: The collection of field data on plant traits is time consuming and this makes it difficult to examine changing patterns of traits along large-scale climate gradients. The present study tests whether trait information derived from regional floras can be used in conjunction with pre existing quadrat data on species presence to derive meaningful relationships between specific morphometric traits and climate. METHODS: Quadrat records were obtained for 867 species in 404 sites from northern China (38-49 degrees N, 82 132 degrees E) together with information on the presence/absence of key traits from floras. Bioclimate parameters for each site were calculated using the BIOME3 model. Principal component analysis and correlation analysis were conducted to determine the most important climate factors. The Akaike Information Criterion was used to select the best relationship between each trait and climate. Canonical correspondence analysis was used to explore the relationships between climate and trait occurrence. KEY RESULTS: The changing abundance of life form, leaf type, phenology, photosynthetic pathway, leaf size and several other morphometric traits are determined by gradients in plant-available moisture (as measured by the ratio of actual to potential evapotranspiration: alpha), growing season temperature (as measured by growing degree-days on a 0 degrees base: GDD(0)) or a combination of these. Different plant functional types (PFTs, as defined by life form, leaf type and phenology) reach maximum abundance in distinct areas of this climate space: for example, evergreen trees occur in the coldest, wettest environments (GDD(0) < 2500 degrees Cd, alpha > 0.38), and deciduous scale-leaved trees occur in drier, warmer environments than deciduous broad-leaved trees. Most leaf-level traits show similar relationships with climate independently of PFT: for example, leaf size in all PFTs increases as the environment becomes wetter and cooler. However, some traits (e.g. petiole length) display different relationships with climate in different PFTs. CONCLUSIONS: Based on presence/absence species data and flora-based trait assignments, the present study demonstrates ecologically plausible trends in the occurrence of key plant traits along climate gradients in northern China. Life form, leaf type, phenology, photosynthetic pathway, leaf size and other key traits reflect climate. The success of these analyses opens the possibility of using quadrat- and flora-based trait analyses to examine climate-trait relationships in other regions of the world. PMID- 19805405 TI - Differential impact of diesel particle composition on pro-allergic dendritic cell function. AB - Diesel exhaust particles (DEP) were described as potent adjuvant in the induction and maintenance of allergic diseases, suggesting that they might play a role in the increase of allergic diseases in the industrialized countries. However, the cellular basis by which these particles enhance allergic immune responses is still a matter of debate. Thus, we exposed immature murine bone marrow-derived dendritic cells (BMDC) to different particles or particle-associated organic compounds in the absence or presence of the maturation stimuli lipopolysaccharide (LPS) and analyzed the cellular maturation, viability, and cytokine production. Furthermore, we monitored the functionality of particle-exposed BMDC to suppress B cell isotype switching to immunoglobulin (Ig) E. Only highly polluted DEP (standard reference material 1650a [SRM1650a]) but not particle-associated organic compounds or less polluted DEP from modern diesel engines were able to modulate the dendritic cell phenotype. SRM1650a particles significantly suppressed LPS-induced IL-12p70 production in murine BMDC, whereas cell-surface marker expression was not altered. Furthermore, SRM1650a-exposed immature BMDC lost the ability to suppress IgE isotype switch in B cells. This study revealed that highly polluted DEP not only interfere with dendritic cell maturation but also additionally with dendritic cell function, thus suggesting a role in T(h)2 immune deviation. PMID- 19805406 TI - Toxicity testing in the 21st century: a view from the chemical industry. PMID- 19805407 TI - Suppression of insulin-like growth factor acid-labile subunit expression--a novel mechanism for deoxynivalenol-induced growth retardation. AB - Consumption of deoxynivalenol (DON), a trichothecene mycotoxin commonly detected in cereal-based foods, causes impaired growth in many animal species. While growth retardation is used as a basis for regulating DON levels in human food, the underlying mechanisms remain poorly understood. Oral exposure of mice to DON rapidly induces multiorgan expression of proinflammatory cytokines, and this is followed by upregulation of several suppressors of cytokine signaling (SOCS), some of which are capable of impairing growth hormone (GH) signaling. The purpose of this study was to test the hypothesis that impairment of the GH axis precedes DON-induced growth retardation in the mouse. Subchronic dietary exposure of young (4-week old) mice to DON (20 ppm) over a period of 2-8 weeks was found to (1) impair weight gain, (2) result in a steady-state plasma DON concentration (40-60 ng/ml), (3) downregulate hepatic insulin-like growth factor acid-labile subunit (IGFALS) mRNA expression, and (4) reduce circulating insulin-like growth factor 1 (IGF1) and IGFALS levels. Acute oral exposure to DON at 0.5-12.5 mg/kg body weight (bw) markedly suppressed hepatic IGFALS mRNA levels within 2 h in a dose dependent fashion, whereas 0.1 mg/kg bw was without effect. DON-induced IGFALS mRNA upregulation occurred both with and without exogenous GH treatment. These latter effects co-occurred with robust hepatic suppressors of cytokine signaling 3 upregulation. Taken together, these data suggest that oral DON exposure perturbs GH axis by suppressing two clinically relevant growth-related proteins, IGFALS and IGF1. Both have potential to serve as biomarkers of effect in populations exposed to this common foodborne mycotoxin. PMID- 19805408 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin inhibits fibroblast growth factor 10-induced prostatic bud formation in mouse urogenital sinus. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) dorsalizes the pattern of prostatic buds developing from the urogenital sinus (UGS) of male fetal mice, causing some buds to form in inappropriate positions while blocking formation of others. This teratogenic TCDD action significantly reduces prostate main duct number and causes ventral prostate agenesis in exposed males. The purpose of this study was to determine whether inhibition of fibroblast growth factor 10 (FGF10) signaling is mechanistically linked to mouse prostatic budding impairment by TCDD. In utero TCDD exposure induced aryl hydrocarbon receptor-responsive cytochrome P450 1b1 messenger RNA (mRNA) in ventral UGS regions where Fgf10 and fibroblast growth factor receptor 2 (Fgfr2) mRNA were expressed and where budding was most severely inhibited by TCDD. However, TCDD exposure did not reduce Fgf10 or Fgfr2 mRNA abundance in the UGS or alter their distribution. Addition of FGF10 protein to UGS organ culture media increased the abundance of UGS basal epithelial cells immunopositive for phosphorylated extracellular signal-regulated kinase (ERK). FGF10 also increased the number of 5-bromo-2'-deoxyuridine (BrdU)-labeled UGS epithelial cells and increased the number of prostatic buds formed per UGS. Addition of TCDD to UGS organ culture media did not alter FGF10-induced ERK activation in UGS basal epithelium but prevented FGF10-induced BrdU incorporation and blocked FGF10-induced prostatic bud formation. These results identify basal urogenital sinus epithelium cells as the key site of FGF10 action during fetal prostate development and suggest that TCDD likely acts downstream of FGFR2 and ERK to restrict UGS epithelial cell proliferation and prevent prostatic bud formation. PMID- 19805409 TI - Weighted feature significance: a simple, interpretable model of compound toxicity based on the statistical enrichment of structural features. AB - In support of the U.S. Tox21 program, we have developed a simple and chemically intuitive model we call weighted feature significance (WFS) to predict the toxicological activity of compounds, based on the statistical enrichment of structural features in toxic compounds. We trained and tested the model on the following: (1) data from quantitative high-throughput screening cytotoxicity and caspase activation assays conducted at the National Institutes of Health Chemical Genomics Center, (2) data from Salmonella typhimurium reverse mutagenicity assays conducted by the U.S. National Toxicology Program, and (3) hepatotoxicity data published in the Registry of Toxic Effects of Chemical Substances. Enrichments of structural features in toxic compounds are evaluated for their statistical significance and compiled into a simple additive model of toxicity and then used to score new compounds for potential toxicity. The predictive power of the model for cytotoxicity was validated using an independent set of compounds from the U.S. Environmental Protection Agency tested also at the National Institutes of Health Chemical Genomics Center. We compared the performance of our WFS approach with classical classification methods such as Naive Bayesian clustering and support vector machines. In most test cases, WFS showed similar or slightly better predictive power, especially in the prediction of hepatotoxic compounds, where WFS appeared to have the best performance among the three methods. The new algorithm has the important advantages of simplicity, power, interpretability, and ease of implementation. PMID- 19805410 TI - Rapamycin inhibits yeast nucleotide excision repair independently of tor kinases. AB - The yeast target of rapamycin (Tor) kinases, Tor1 and Tor2, belong to the phosphatidylinositol 3-kinase-related family of proteins, which are involved in the cellular response to DNA damage and changes in nutrient conditions. In contrast to yeast, many eukaryotes possess a single Tor kinase. Regardless of the number of Tor kinases in an organism, two distinct complexes involving Tor proteins exist in eukaryotes, TORC1 and TORC2. The yeast TORC1, containing Tor1 or Tor2, is sensitive to the antibiotic rapamycin. The yeast TORC2 is insensitive to rapamycin. We examined the influence of rapamycin treatment upon yeast transcription-coupled nucleotide excision repair in a gene transcribed by RNA polymerase II. We also examined tor mutants for their ability to perform transcription-coupled repair in the absence or presence of rapamycin. Ostensibly lacking TORC1 and TORC2 function, a tor1tor2(ts) mutant grown at the nonpermissive temperature exhibited similar rates of repair as the wild-type strain. However, repair of both strands in genes decreases in the wild-type strain and the tor1tor2(ts) mutant exposed to rapamycin. Rapamycin may be inhibiting DNA repair independently of the Tor kinases. In yeast, FPR1 encodes the rapamycin-binding protein Fpr1 that inhibits the TORC1 kinase in the presence of rapamycin. Fap1 competes with rapamycin for Fpr1 binding. Deletion of the FPR1 or FAP1 gene abolishes the inhibitory effect of rapamycin on repair. Thus, the decreased repair observed following rapamycin treatment is independent of TORC1/2 function and likely due to a function of Fap1. We suggest that Fap1 and peptidyl prolyl isomerases, particularly Fpr1, function in the cellular response to genotoxic stress. Our findings have clinical implications for genetic toxicities associated with genotoxic agents when coadministered with rapamycin. PMID- 19805411 TI - Unusual localization of a malignant fibrous histiocytoma on the mitral valve. AB - The present report describes the case of a 55-year-old woman who suffered from cardio-embolic stroke originating from malignant fibrous histiocytoma (MFH) localized on the mitral valve. The patient underwent transthoracic two-/three dimensional and transoesophageal echocardiography which demonstrated the mass protruding in the outflow tract of the left ventricle. Differential diagnosis had to be made with other masses in the left ventricle, such as thrombi, vegetations, and cardiac tumours. Surgery was performed to remove the tumour and the surgery findings confirmed echocardiographic images. Primary cardiac tumours are a rare entity, and their incidence is approximately 0.0017-0.019%. The majority of them are benign, but in a quarter of cases they are malignant. This case is an example of an MFH which caused embolism to the central nervous system. PMID- 19805412 TI - Mitral valve disease as well as uncommon extensive epipericardial and intramyocardial calcification secondary to massive mitral annular calcification. AB - A 71-year-old woman with a history of childhood pulmonary tuberculosis was admitted to our hospital for exertional dyspnoea (NYHA functional class II). Transthoracic and transoesophageal echocardiography demonstrated moderate to severe mixed mitral valve disease due to massive mitral annular calcification (MAC) and extensive infiltrative calcification of the atrioventricular groove. In addition, a very uncommon intramyocardial calcification of the ventricular septum and the lateral free wall was diagnosed. This case demonstrates a rare combination of mitral valve disease secondary to MAC, and a small hypertrophied left ventricle, as well as epipericardial and myocardial calcification likely due either to the massive MAC with myocardial extension or to former tuberculous perimyocarditis. The multidimensional imaging approach, which has been used in this particularly case, provided an excellent visualization and clinical evaluation of this rare finding. PMID- 19805413 TI - Real-time three-dimensional transoesophageal echocardiography in the assessment of aortic valve stenosis. AB - AIMS: To determine the feasibility of real-time three-dimensional transoesophageal echocardiography (3D-TOE) in the evaluation of aortic valve stenosis, to study its reliability, and to test the concordance of this new method when compared with transthoracic two-dimensional echocardiography (2D-TTE) as the diagnostic standard. METHODS AND RESULTS: Fifty-nine consecutive patients with moderate-to-severe aortic valve stenosis were assessed by means of 2D-TTE and 3D-TOE by independent blinded observers. Aortic valve planimetry was possible in 94.9% of patients. Inter-observer intraclass correlation coefficients (ICC) were 0.892 (CI 95% 0.818-0.936; P < 0.001), and 0.871 (CI 95% 0.780-0.925; P < 0.001) for 2D-TTE and 3D-TOE, respectively. Bland-Altman plot showed a mean difference in aortic valve area (AVA) of 0.040 cm(2), with 2D-TTE yielding larger values than 3D-TOE. ICC of both methods was 0.724 (CI 95% 0.530-0.839; P < 0.001). CONCLUSION: Assessment of AVA by means of 3D-TOE is feasible in most patients with aortic valve stenosis. Reliability of the measurement is good. However, there is some disagreement with standard 2D-TTE that needs further investigation. PMID- 19805414 TI - Home improvements: how the malaria parasite makes the red blood cell home sweet home. AB - A recent study published in Nature by de Koning-Ward et al. provides further insights into the way in which the lethal malaria parasite, Plasmodium falciparum, remodels the host red blood cell during the pathogenic blood stage of development. PMID- 19805415 TI - Rapamycin: the cure for all that ails. AB - Target of rapamycin (TOR) signaling stimulates cell growth by regulating protein synthesis in response to a variety of stimuli in a wide range of species and is inhibited by rapamycin, a naturally occurring antifungal compound produced by bacteria and discovered on Easter Island or in the local vernacular, Rapa Nui (rapamycin's namesake). Recently, rapamycin was shown to extend life span for mice, even when administered late in life, suggesting that inhibiting the mammalian TOR pathway may improve health span for people. PMID- 19805416 TI - Identity crisis when telomeres left unprotected. AB - Loss of shelterin components TRF2 or POT1a-TPP1 complex from the chromosome end triggers DNA damage response (DDR) and aberrant DNA repair events. In a recent Nature paper, Chang and colleagues reported that the DNA repair protein Mre11 contributes to multiple events at the uncapped telomere, including ataxia telangiectasia-mutated (ATM)-dependent signaling, processing of the telomeric G tail and homologous recombination (HR). PMID- 19805417 TI - Motor cortex plasticity predicts recovery in acute stroke. AB - Repetitive transcranial magnetic stimulation of the brain given as intermittent theta burst stimulation (iTBS) can induce long-term potentiation (LTP)-like changes in the stimulated hemisphere and long-term depression (LTD)-like changes in the opposite hemisphere. We evaluated whether LTP- and LTD-like changes produced by iTBS in acute stroke correlate with outcome at 6 months. We evaluated the excitability of affected hemisphere (AH) and unaffected hemisphere (UH) by measuring motor threshold and motor-evoked potential (MEP) amplitude under baseline conditions and after iTBS of AH in 17 patients with acute ischemic stroke. Baseline amplitude of MEPs elicited from AH was significantly smaller than that of MEPs elicited from UH, and baseline motor threshold was higher for the AH. Higher baseline MEP values in UH correlated with poor prognosis. iTBS produced a significant increase in MEP amplitude for AH that was significantly correlated with recovery. A nonsignificant decrease in MEP amplitude was observed for the UH. When the decrease in the amplitude of UH MEPs was added to the regression model, the correlation was even higher. Functional recovery is directly correlated with LTP-like changes in AH and LTD-like changes in UH and inversely correlated with the baseline excitability of UH. PMID- 19805418 TI - Does brain activity at rest reflect adaptive strategies? Evidence from speech processing after cochlear implantation. AB - In functional neuroimaging studies, task-related activity refers to the signal difference between the stimulation and rest conditions. We asked whether long term changes in the sensory environment may affect brain activity at rest. To answer this question, we compared regional cerebral blood flow between a group of normally hearing controls and a group of cochlear-implanted (CI) deaf patients. Here we present evidence that long-term alteration of auditory experience, such as profound deafness followed by partial auditory recuperation through cochlear implantation, leads to functional cortical reorganizations at rest. Without any visual or auditory stimulation, CI subjects showed changes of cerebral blood flow in the visual, auditory cortex, Broca area, and in the posterior temporal cortex with an increment of activity in these areas from the time of activation of the implant to less than a year after the implantation. PMID- 19805419 TI - Ventral premotor and inferior parietal cortices make distinct contribution to action organization and intention understanding. AB - It is well known that ventral premotor area F5 codes the goal of executed and observed motor acts. This area is anatomically connected with part of the inferior parietal cortex (area PFG), which has been recently shown to play a role in action organization and intention understanding. The aims of the present study were 1) to assess whether the discharge of F5 motor neurons and mirror neurons (MNs) codes action goals and 2) to clarify the relative contribution of F5 and PFG in action organization and intention understanding. To this purpose, we first recorded from F5 motor neurons and MNs of 2 monkeys while performing a motor task constituted by 2 actions ("grasp-to-eat" and "grasp-to-place") or observing the same task done by an experimenter. Results showed that some F5 neurons code grasping according to the goal of the action in which it is embedded. Subsequently, we recorded from PFG motor neurons and MNs of the same monkeys, using the same tasks. The comparison between the neuronal properties of F5 and PFG motor neurons suggests that PFG plays a major role in organizing natural actions. Furthermore, the similarities between MNs properties of the 2 areas indicate that they constitute a functional circuit underlying others' intention understanding. PMID- 19805421 TI - The evolution of society. PMID- 19805420 TI - Age-related shifts in brain activity dynamics during task switching. AB - Cognitive aging studies have suggested that older adults show declines in both sustained and transient cognitive control processes. However, previous neuroimaging studies have primarily focused on age-related change in the magnitude, but not temporal dynamics, of brain activity. The present study compared brain activity dynamics in healthy old and young adults during task switching. A mixed blocked/event-related functional magnetic resonance imaging design enabled separation of transient and sustained neural activity associated with cognitive control. Relative to young adults, older adults exhibited not only decreased sustained activity in the anterior prefrontal cortex (aPFC) during task switching blocks but also increased transient activity on task-switch trials. Another pattern of age-related shift in dynamics was present in the lateral PFC (lPFC) and posterior parietal cortex (PPC), with younger adults showing a cue related response during task-switch trials in lPFC and PPC, whereas older adults exhibited switch-related activation during the cue period in PPC only. In all 3 regions, these qualitatively distinct patterns of brain activity predicted qualitatively distinct patterns of behavioral performance across the 2 age groups. Together, these results suggest that older adults may shift from a proactive to reactive cognitive control strategy as a means of retaining relatively preserved behavioral performance in the face of age-related neurocognitive changes. PMID- 19805422 TI - Formalizing Darwinism and inclusive fitness theory. AB - Inclusive fitness maximization is a basic building block for biological contributions to any theory of the evolution of society. There is a view in mathematical population genetics that nothing is caused to be maximized in the process of natural selection, but this is explained as arising from a misunderstanding about the meaning of fitness maximization. Current theoretical work on inclusive fitness is discussed, with emphasis on the author's 'formal Darwinism project'. Generally, favourable conclusions are drawn about the validity of assuming fitness maximization, but the need for continuing work is emphasized, along with the possibility that substantive exceptions may be uncovered. The formal Darwinism project aims more ambitiously to represent in a formal mathematical framework the central point of Darwin's Origin of Species, that the mechanical processes of inheritance and reproduction can give rise to the appearance of design, and it is a fitting ambition in Darwin's bicentenary year to capture his most profound discovery in the lingua franca of science. PMID- 19805423 TI - Beyond society: the evolution of organismality. AB - The evolution of organismality is a social process. All organisms originated from groups of simpler units that now show high cooperation among the parts and are nearly free of conflicts. We suggest that this near-unanimous cooperation be taken as the defining trait of organisms. Consistency then requires that we accept some unconventional organisms, including some social insect colonies, some microbial groups and viruses, a few sexual partnerships and a number of mutualistic associations. Whether we call these organisms or not, a major task is to explain such cooperative entities, and our survey suggests that many of the traits commonly used to define organisms are not essential. These non-essential traits include physical contiguity, indivisibility, clonality or high relatedness, development from a single cell, short-term and long-term genetic cotransmission, germ-soma separation and membership in the same species. PMID- 19805424 TI - Social evolution in micro-organisms and a Trojan horse approach to medical intervention strategies. AB - Medical science is typically pitted against the evolutionary forces acting upon infective populations of bacteria. As an alternative strategy, we could exploit our growing understanding of population dynamics of social traits in bacteria to help treat bacterial disease. In particular, population dynamics of social traits could be exploited to introduce less virulent strains of bacteria, or medically beneficial alleles into infective populations. We discuss how bacterial strains adopting different social strategies can invade a population of cooperative wild type, considering public good cheats, cheats carrying medically beneficial alleles (Trojan horses) and cheats carrying allelopathic traits (anti-competitor chemical bacteriocins or temperate bacteriophage viruses). We suggest that exploitation of the ability of cheats to invade cooperative, wild-type populations is a potential new strategy for treating bacterial disease. PMID- 19805425 TI - The evolution of extreme altruism and inequality in insect societies. AB - In eusocial organisms, some individuals specialize in reproduction and others in altruistic helping. The evolution of eusociality is, therefore, also the evolution of remarkable inequality. For example, a colony of honeybees (Apis mellifera) may contain 50 000 females all of whom can lay eggs. But 100 per cent of the females and 99.9 per cent of the males are offspring of the queen. How did such extremes evolve? Phylogenetic analyses show that high relatedness was almost certainly necessary for the origin of eusociality. However, even the highest family levels of kinship are insufficient to cause the extreme inequality seen in e.g. honeybees via 'voluntary altruism'. 'Enforced altruism' is needed, i.e. social pressures that deter individuals from attempting to reproduce. Coercion acts at two stages in an individual's life cycle. Queens are typically larger so larvae can be coerced into developing into workers by being given less food. Workers are coerced into working by 'policing', in which workers or the queen eat worker-laid eggs or aggress fertile workers. In some cases, individuals rebel, such as when stingless bee larvae develop into dwarf queens. The incentive to rebel is strong as an individual is the most closely related to its own offspring. However, because individuals gain inclusive fitness by rearing relatives, there is also a strong incentive to 'acquiesce' to social coercion. In a queenright honeybee colony, the policing of worker-laid eggs is very effective, which results in most workers working instead of attempting to reproduce. Thus, extreme altruism is due to both kinship and coercion. Altruism is frequently seen as a Darwinian puzzle but was not a puzzle that troubled Darwin. Darwin saw his difficulty in explaining how individuals that did not reproduce could evolve, given that natural selection was based on the accumulation of small heritable changes. The recognition that altruism is an evolutionary puzzle, and the solution was to wait another 100 years for William Hamilton. PMID- 19805426 TI - Social stability and helping in small animal societies. AB - In primitively eusocial societies, all individuals can potentially reproduce independently. The key fact that we focus on in this paper is that individuals in such societies instead often queue to inherit breeding positions. Queuing leads to systematic differences in expected future fitness. We first discuss the implications this has for variation in behaviour. For example, because helpers nearer to the front of the queue have more to lose, they should work less hard to rear the dominant's offspring. However, higher rankers may be more aggressive than low rankers, even if they risk injury in the process, if aggression functions to maintain or enhance queue position. Second, we discuss how queuing rules may be enforced through hidden threats that rarely have to be carried out. In fishes, rule breakers face the threat of eviction from the group. In contrast, subordinate paper wasps are not injured or evicted during escalated challenges against the dominant, perhaps because they are more valuable to the dominant. We discuss evidence that paper-wasp dominants avoid escalated conflicts by ceding reproduction to subordinates. Queuing rules appear usually to be enforced by individuals adjacent in the queue rather than by dominants. Further manipulative studies are required to reveal mechanisms underlying queue stability and to elucidate what determines queue position in the first place. PMID- 19805427 TI - Lifetime monogamy and the evolution of eusociality. AB - All evidence currently available indicates that obligatory sterile eusocial castes only arose via the association of lifetime monogamous parents and offspring. This is consistent with Hamilton's rule (br(s) > r(o)c), but implies that relatedness cancels out of the equation because average relatedness to siblings (r(s)) and offspring (r(o)) are both predictably 0.5. This equality implies that any infinitesimally small benefit of helping at the maternal nest (b), relative to the cost in personal reproduction (c) that persists throughout the lifespan of entire cohorts of helpers suffices to establish permanent eusociality, so that group benefits can increase gradually during, but mostly after the transition. The monogamy window can be conceptualized as a singularity comparable with the single zygote commitment of gametes in eukaryotes. The increase of colony size in ants, bees, wasps and termites is thus analogous to the evolution of multicellularity. Focusing on lifetime monogamy as a universal precondition for the evolution of obligate eusociality simplifies the theory and may help to resolve controversies about levels of selection and targets of adaptation. The monogamy window underlines that cooperative breeding and eusociality are different domains of social evolution, characterized by different sectors of parameter space for Hamilton's rule. PMID- 19805428 TI - Adaptation and the genetics of social behaviour. AB - In recent years much progress has been made towards understanding the selective forces involved in the evolution of social behaviour including conflicts over reproduction among group members. Here, I argue that an important additional step necessary for advancing our understanding of the resolution of potential conflicts within insect societies is to consider the genetics of the behaviours involved. First, I discuss how epigenetic modifications of behaviour may affect conflict resolution within groups. Second, I review known natural polymorphisms of social organization to demonstrate that a lack of consideration of the genetic mechanisms involved may lead to erroneous explanations of the adaptive significance of behaviour. Third, I suggest that, on the basis of recent genetic studies of sexual conflict in Drosophila, it is necessary to reconsider the possibility of within-group manipulation by means of chemical substances (i.e. pheromones). Fourth, I address the issue of direct versus indirect genetic effects, which is of particular importance for the study of behaviour in social groups. Fifth, I discuss the issue of how a genetic influence on dominance hierarchies and reproductive division of labour can have secondary effects, for example in the evolution of promiscuity. Finally, because the same sets of genes (e.g. those implicated in chemical signalling and the responses that are triggered) may be used even in species as divergent as ants, cooperative breeding birds and primates, an integration of genetic mechanisms into the field of social evolution may also provide unifying ideas. PMID- 19805429 TI - The evolution of cooperative breeding in birds: kinship, dispersal and life history. AB - The evolution of cooperation among animals has posed a major problem for evolutionary biologists, and despite decades of research into avian cooperative breeding systems, many questions about the evolution of their societies remain unresolved. A review of the kin structure of avian societies shows that a large majority live in kin-based groups. This is consistent with the proposed evolutionary routes to cooperative breeding via delayed dispersal leading to family formation, or limited dispersal leading to kin neighbourhoods. Hypotheses proposed to explain the evolution of cooperative breeding systems have focused on the role of population viscosity, induced by ecological/demographic constraints or benefits of philopatry, in generating this kin structure. However, comparative analyses have failed to generate robust predictions about the nature of those constraints, nor differentiated between the viscosity of social and non-social populations, except at a coarse level. I consider deficiencies in our understanding of how avian dispersal strategies differ between social and non social species, and suggest that research has focused too narrowly on population viscosity and that a broader perspective that encompasses life history and demographic processes may provide fresh insights into the evolution of avian societies. PMID- 19805430 TI - Structure and function in mammalian societies. AB - Traditional interpretations of the evolution of animal societies have suggested that their structure is a consequence of attempts by individuals to maximize their inclusive fitness within constraints imposed by their social and physical environments. In contrast, some recent re-interpretations have argued that many aspects of social organization should be interpreted as group-level adaptations maintained by selection operating between groups or populations. Here, I review our current understanding of the evolution of mammalian societies, focusing, in particular, on the evolution of reproductive strategies in societies where one dominant female monopolizes reproduction in each group and her offspring are reared by other group members. Recent studies of the life histories of females in these species show that dispersing females often have little chance of establishing new breeding groups and so are likely to maximize their inclusive fitness by helping related dominants to rear their offspring. As in eusocial insects, increasing group size can lead to a progressive divergence in the selection pressures operating on breeders and helpers and to increasing specialization in their behaviour and life histories. As yet, there is little need to invoke group-level adaptations in order to account for the behaviour of individuals or the structure of mammalian groups. PMID- 19805431 TI - Nepotistic cooperation in non-human primate groups. AB - Darwin was struck by the many similarities between humans and other primates and believed that these similarities were the product of common ancestry. He would be even more impressed by the similarities if he had known what we have learned about primates over the last 50 years. Genetic kinship has emerged as the primary organizing force in the evolution of primate social organization and the patterning of social behaviour in non-human primate groups. There are pronounced nepotistic biases across the primate order, from tiny grey mouse lemurs (Microcebus murinus) that forage alone at night but cluster with relatives to sleep during the day, to cooperatively breeding marmosets that rely on closely related helpers to rear their young, rhesus macaque (Macaca mulatta) females who acquire their mother's rank and form strict matrilineal dominance hierarchies, male howler monkeys that help their sons maintain access to groups of females and male chimpanzees (Pan troglodytes) that form lasting relationships with their brothers. As more evidence of nepotism has accumulated, important questions about the evolutionary processes underlying these kin biases have been raised. Although kin selection predicts that altruism will be biased in favour of relatives, it is difficult to assess whether primates actually conform to predictions derived from Hamilton's rule: br > c. In addition, other mechanisms, including contingent reciprocity and mutualism, could contribute to the nepotistic biases observed in non-human primate groups. There are good reasons to suspect that these processes may complement the effects of kin selection and amplify the extent of nepotistic biases in behaviour. PMID- 19805432 TI - Evolving the ingredients for reciprocity and spite. AB - Darwin never provided a satisfactory account of altruism, but posed the problem beautifully in light of the logic of natural selection. Hamilton and Williams delivered the necessary satisfaction by appealing to kinship, and Trivers showed that kinship was not necessary as long as the originally altruistic act was conditionally reciprocated. From the late 1970s to the present, the kinship theories in particular have been supported by considerable empirical data and elaborated to explore a number of other social interactions such as cooperation, selfishness and punishment, giving us what is now a rich description of the nature of social relationships among organisms. There are, however, two forms of theoretically possible social interactions-reciprocity and spite-that appear absent or nearly so in non-human vertebrates, despite considerable research efforts on a wide diversity of species. We suggest that the rather weak comparative evidence for these interactions is predicted once we consider the requisite socioecological pressures and psychological mechanisms. That is, a consideration of ultimate demands and proximate prerequisites leads to the prediction that reciprocity and spite should be rare in non-human animals, and common in humans. In particular, reciprocity and spite evolved in humans because of adaptive demands on cooperation among unrelated individuals living in large groups, and the integrative capacities of inequity detection, future-oriented decision-making and inhibitory control. PMID- 19805433 TI - The ecology of social transitions in human evolution. AB - We know that there are fundamental differences between humans and living apes, and also between living humans and their extinct relatives. It is also probably the case that the most significant and divergent of these differences relate to our social behaviour and its underlying cognition, as much as to fundamental differences in physiology, biochemistry or anatomy. In this paper, we first attempt to demarcate what are the principal differences between human and other societies in terms of social structure, organization and relationships, so that we can identify what derived features require explanation. We then consider the evidence of the archaeological and fossil record, to determine the most probable context in time and taxonomy, of these evolutionary trends. Finally, we attempt to link five major transitional points in hominin evolution to the selective context in which they occurred, and to use the principles of behavioural ecology to understand their ecological basis. Critical changes in human social organization relate to the development of a larger scale of fission and fusion; the development of a greater degree of nested substructures within the human community; and the development of intercommunity networks. The underlying model that we develop is that the evolution of 'human society' is underpinned by ecological factors, but these are influenced as much by technological and behavioural innovations as external environmental change. PMID- 19805434 TI - Culture and the evolution of human cooperation. AB - The scale of human cooperation is an evolutionary puzzle. All of the available evidence suggests that the societies of our Pliocene ancestors were like those of other social primates, and this means that human psychology has changed in ways that support larger, more cooperative societies that characterize modern humans. In this paper, we argue that cultural adaptation is a key factor in these changes. Over the last million years or so, people evolved the ability to learn from each other, creating the possibility of cumulative, cultural evolution. Rapid cultural adaptation also leads to persistent differences between local social groups, and then competition between groups leads to the spread of behaviours that enhance their competitive ability. Then, in such culturally evolved cooperative social environments, natural selection within groups favoured genes that gave rise to new, more pro-social motives. Moral systems enforced by systems of sanctions and rewards increased the reproductive success of individuals who functioned well in such environments, and this in turn led to the evolution of other regarding motives like empathy and social emotions like shame. PMID- 19805435 TI - The evolutionary and ecological roots of human social organization. AB - Social organization among human foragers is characterized by a three-generational system of resource provisioning within families, long-term pair-bonding between men and women, high levels of cooperation between kin and non-kin, and relatively egalitarian social relationships. In this paper, we suggest that these core features of human sociality result from the learning- and skill-intensive human foraging niche, which is distinguished by a late age-peak in caloric production, high complementarity between male and female inputs to offspring viability, high gains to cooperation in production and risk-reduction, and a lack of economically defensible resources. We present an explanatory framework for understanding variation in social organization across human societies, highlighting the interactive effects of four key ecological and economic variables: (i) the role of skill in resource production; (ii) the degree of complementarity in male and female inputs into production; (iii) economies of scale in cooperative production and competition; and (iv) the economic defensibility of physical inputs into production. Finally, we apply this framework to understanding variation in social and political organization across foraging, horticulturalist, pastoralist and agriculturalist societies. PMID- 19805436 TI - Trust and cooperation among economic agents. AB - The units that are subject to selection pressure in evolutionary biology are 'strategies', which are conditional actions ('Do P if X occurs, otherwise do Q'). In contrast, the units in economics select strategies from available menus so as to further their projects and purposes. As economic agents do not live in isolation, each agent's optimum choice, in general, depends on the choices made by others. Because their projects and purposes involve the future, not just the present, each agent reasons about the likely present and future consequences of their respective choices. That is why beliefs, about what others may do and what the consequences of those choices could be, are at the basis of strategy selection. A catalogue of social environments is constructed in which agents not only promise each other's cooperation, but also rationally believe that the promises will be kept. Unfortunately, non-cooperation arising from mistrust can be the outcome in those same environments: societies harbour multiple 'equilibria' and can skid from cooperation to non-cooperation. Moreover, a pre occupation among analysts with the Prisoners' Dilemma game has obscured the fact that cooperative arrangements can harbour not only inequality, but also exploitation. The analysis is used to discuss why international cooperation over the use of global public goods has proved to be so elusive. PMID- 19805437 TI - Isoform divergence of the filamin family of proteins. AB - The vertebrate filamin family (A, B, and C) is part of the spectrin family of actin cross-linking proteins. Family members share high sequence similarity (>64%) and have both common and isoform-distinct functionalities. To identify the basis for isoform-specific functionality, we perform an evolutionary trace of chordate filamin at the granularity of single residues. Our trace methodology is constrained to focus on neofunctionality by requiring that one isoform remain the ancestral type, whereas at least one isoform has an accepted mutation. We call divergence meeting these characteristics "class-distinctive." To obtain a temporal and spatial context for class-distinctive residues, we derive an all atom model of full-length filamin A by homology modeling and joining individual domains. We map onto our model both conserved and class-distinctive residues along with the period (Teleostei, Amphibian, and Mammalian) in which they diverged. Our phylogenetic analysis suggests that filamins diverged from a common ancestral gene between urochordate and vertebrate lineages. Filamins also diverged the most just after gene duplication, in the Teleostei period, with filamin C remaining closest to ancestral filamin. At the residue level, domains with well-characterized interfaces, IgFLN 17 and IgFLN 21 (immunoglobulin, Ig), have diverged in potentially critical residues in their adhesion protein-binding interfaces, signifying that isoforms may bind or regulate ligand binding differentially. Similarly, isoform divergence in a region associated with F actin binding regulation suggests that isoforms differentially regulate F-actin binding. In addition, we observe some class-distinctive residues in the vicinity of missense mutations that cause filamin A and B-associated skeletal disorders. Our analysis, utilizing both spatial and temporal granularity, has identified potentially important residues responsible for vertebrate filamin isoform specific divergence-significantly in regions where few binding partners have been discovered to date- and suggests yet to be discovered filamin-binding partners and isoform-specific differential regulation with these binding partners. PMID- 19805438 TI - A statistical evaluation of models for the initial settlement of the american continent emphasizes the importance of gene flow with Asia. AB - Although there is agreement in that the Bering Strait was the entry point for the initial colonization of the American continent, there is considerable uncertainty regarding the timing and pattern of human migration from Asia to America. In order to perform a statistical assessment of the relative probability of alternative migration scenarios and to estimate key demographic parameters associated with them, we used an approximate Bayesian computation framework to analyze a data set of 401 autosomal microsatellite loci typed in 29 native American populations. A major finding is that a single, discrete, wave of colonization is highly inconsistent with observed levels of genetic diversity. A scenario with two discrete migration waves is also not supported by the data. The current genetic diversity of Amerindian populations is best explained by a third model involving recurrent gene flow between Asia and America, after initial colonization. We estimate that this colonization involved about 100 individuals and occurred some 13,000 years ago, in agreement with well-established archeological data. PMID- 19805439 TI - Phylogenetic distributions and histories of proteins involved in anaerobic pyruvate metabolism in eukaryotes. AB - Protists that live in low oxygen conditions often oxidize pyruvate to acetate via anaerobic ATP-generating pathways. Key enzymes that commonly occur in these pathways are pyruvate:ferredoxin oxidoreductase (PFO) and [FeFe]-hydrogenase (H(2)ase) as well as the associated [FeFe]-H(2)ase maturase proteins HydE, HydF, and HydG. Determining the origins of these proteins in eukaryotes is of key importance to understanding the origins of anaerobic energy metabolism in microbial eukaryotes. We conducted a comprehensive search for genes encoding these proteins in available whole genomes and expressed sequence tag data from diverse eukaryotes. Our analyses of the presence/absence of eukaryotic PFO, [FeFe]-H(2)ase, and H(2)ase maturase sequences across eukaryotic diversity reveal orthologs of these proteins encoded in the genomes of a variety of protists previously not known to contain them. Our phylogenetic analyses revealed: 1) extensive lateral gene transfers of both PFO and [FeFe]-H(2)ase in eubacteria, 2) decreased support for the monophyly of eukaryote PFO domains, and 3) that eukaryotic [FeFe]-H(2)ases are not monophyletic. Although there are few eukaryote [FeFe]-H(2)ase maturase orthologs characterized, phylogenies of these proteins do recover eukaryote monophyly, although a consistent eubacterial sister group for eukaryotic homologs could not be determined. An exhaustive search for these five genes in diverse genomes from two representative eubacterial groups, the Clostridiales and the alpha-proteobacteria, shows that although these enzymes are nearly universally present within the former group, they are very rare in the latter. No alpha-proteobacterial genome sequenced to date encodes all five proteins. Molecular phylogenies and the extremely restricted distribution of PFO, [FeFe]-H(2)ases, and their associated maturases within the alpha-proteobacteria do not support a mitochondrial origin for these enzymes in eukaryotes. However, the unexpected prevalence of PFO, pyruvate:NADP oxidoreductase, [FeFe]-H(2)ase, and the maturase proteins encoded in genomes of diverse eukaryotes indicates that these enzymes have an important role in the evolution of microbial eukaryote energy metabolism. PMID- 19805440 TI - Chordate Hox and ParaHox gene clusters differ dramatically in their repetitive element content. AB - The ParaHox and Hox gene clusters control aspects of animal anterior-posterior development and are related as paralogous evolutionary sisters. Despite this relationship, it is not clear if the clusters operate in similar ways, with similar constraints. To compare clusters, we examined the transposable-element (TE) content of amphioxus and mammalian ParaHox and Hox clusters. Chordate Hox clusters are known to be largely devoid of TEs, possibly due to gene regulation and constraints on clustering in these animals. Here, we describe several novel amphioxus TEs and show that the amphioxus ParaHox cluster is a hotspot for TE insertion. TE contents of mammalian ParaHox loci are at background levels, in stark contrast to chordate Hox clusters. This marks a significant difference between Hox and ParaHox clusters. The presence of so many potentially disruptive elements implies selection constrains these ParaHox clusters as they have not dispersed despite 500 My of evolution for each lineage. PMID- 19805441 TI - Serum levels of CXCL13 are elevated in active multiple sclerosis. AB - There is increasing recognition of the important role that B cells play in the pathogenesis of multiple sclerosis (MS). Recently it was reported that the B cell chemokine CXCL13 is elevated in MS serum and cerebrospinal fluid. Here we study whether serum levels of CXCL13 are associated with active MS. We measured serum levels of CXCL13 by enzyme-linked immunosorbent assay in 74 patients with relapsing MS randomized to interferon beta 1b or glatiramer acetate and examined with monthly 3 T brain MRI scans optimized for detection of gadolinium enhancement for up to 2 years. The median (range) serum levels of CXCL13 pre treatment were 40 (3-171) pg/ml. Serum levels of CXCL13 were significantly higher at times of active brain MRI scans (p < 0.01). Furthermore, serum levels were higher in patients who never reached MRI remission compared with those in complete (p < 0.01) or partial (p = 0.01) remission. There was a significant positive correlation between the pattern of serum levels of CXCL13 and MRI activity during the first (r = 0.33, p < 0.05) and the full 2 years (r = 0.35, p < 0.01) of the study. Treatment with interferon beta 1b or glatiramer acetate did not affect serum CXCL13. We conclude that the serum levels of the B cell chemokine CXCL13 are associated with active MS. PMID- 19805442 TI - High prevalence of multiple sclerosis in the Swedish county of Varmland. AB - Previous epidemiological studies have indicated that the county of Varmland in western Sweden may be a high-risk zone for multiple sclerosis (MS). The objective of this study was to determine the prevalence in the area. Hospital and general practice medical files were scrutinized. The diagnostic criteria of Poser were used, with 31 December 2002 as prevalence day. The prevalence was 170.07 per 100,000 inhabitants. The average annual incidence was 6.39 to 6.46 per 100,000 (1991-1995, 1996-2000). Multiple sclerosis was 2.3 times more common among women than men. There was a variation in prevalence among the 16 municipalities, however it was not statistically significant. The rates seemed highest in the southwestern part of the county, roughly similar in location to findings some 70 years earlier. When the prevalence ratios by geographical units for the county in 1933 were applied to the current prevalence, the distribution from these estimated cases differed from homogeneity with very high significance (p < 0.00001 ). In conclusion, this study supports previous reports indicating that Varmland continues to be a high-risk zone for MS and shares in the diffusion of the disease at the county level which we had presented for the country as a whole. PMID- 19805443 TI - Statistical challenges of high-dimensional data. AB - Modern applications of statistical theory and methods can involve extremely large datasets, often with huge numbers of measurements on each of a comparatively small number of experimental units. New methodology and accompanying theory have emerged in response: the goal of this Theme Issue is to illustrate a number of these recent developments. This overview article introduces the difficulties that arise with high-dimensional data in the context of the very familiar linear statistical model: we give a taste of what can nevertheless be achieved when the parameter vector of interest is sparse, that is, contains many zero elements. We describe other ways of identifying low-dimensional subspaces of the data space that contain all useful information. The topic of classification is then reviewed along with the problem of identifying, from within a very large set, the variables that help to classify observations. Brief mention is made of the visualization of high-dimensional data and ways to handle computational problems in Bayesian analysis are described. At appropriate points, reference is made to the other papers in the issue. PMID- 19805444 TI - Selective inference in complex research. AB - We explain the problem of selective inference in complex research using a recently published study: a replicability study of the associations in order to reveal and establish risk loci for type 2 diabetes. The false discovery rate approach to such problems will be reviewed, and we further address two problems: (i) setting confidence intervals on the size of the risk at the selected locations and (ii) selecting the replicable results. PMID- 19805445 TI - Observed universality of phase transitions in high-dimensional geometry, with implications for modern data analysis and signal processing. AB - We review connections between phase transitions in high-dimensional combinatorial geometry and phase transitions occurring in modern high-dimensional data analysis and signal processing. In data analysis, such transitions arise as abrupt breakdown of linear model selection, robust data fitting or compressed sensing reconstructions, when the complexity of the model or the number of outliers increases beyond a threshold. In combinatorial geometry, these transitions appear as abrupt changes in the properties of face counts of convex polytopes when the dimensions are varied. The thresholds in these very different problems appear in the same critical locations after appropriate calibration of variables. These thresholds are important in each subject area: for linear modelling, they place hard limits on the degree to which the now ubiquitous high-throughput data analysis can be successful; for robustness, they place hard limits on the degree to which standard robust fitting methods can tolerate outliers before breaking down; for compressed sensing, they define the sharp boundary of the undersampling/sparsity trade-off curve in undersampling theorems. Existing derivations of phase transitions in combinatorial geometry assume that the underlying matrices have independent and identically distributed Gaussian elements. In applications, however, it often seems that Gaussianity is not required. We conducted an extensive computational experiment and formal inferential analysis to test the hypothesis that these phase transitions are universal across a range of underlying matrix ensembles. We ran millions of linear programs using random matrices spanning several matrix ensembles and problem sizes; visually, the empirical phase transitions do not depend on the ensemble, and they agree extremely well with the asymptotic theory assuming Gaussianity. Careful statistical analysis reveals discrepancies that can be explained as transient terms, decaying with problem size. The experimental results are thus consistent with an asymptotic large-n universality across matrix ensembles; finite-sample universality can be rejected. PMID- 19805447 TI - An overview of recent developments in genomics and associated statistical methods. AB - The landscape of genomics has changed drastically in the last two decades. Increasingly inexpensive sequencing has shifted the primary focus from the acquisition of biological sequences to the study of biological function. Assays have been developed to study many intricacies of biological systems, and publicly available databases have given rise to integrative analyses that combine information from many sources to draw complex conclusions. Such research was the focus of the recent workshop at the Isaac Newton Institute, 'High dimensional statistics in biology'. Many computational methods from modern genomics and related disciplines were presented and discussed. Using, as much as possible, the material from these talks, we give an overview of modern genomics: from the essential assays that make data-generation possible, to the statistical methods that yield meaningful inference. We point to current analytical challenges, where novel methods, or novel applications of extant methods, are presently needed. PMID- 19805446 TI - On landmark selection and sampling in high-dimensional data analysis. AB - In recent years, the spectral analysis of appropriately defined kernel matrices has emerged as a principled way to extract the low-dimensional structure often prevalent in high-dimensional data. Here, we provide an introduction to spectral methods for linear and nonlinear dimension reduction, emphasizing ways to overcome the computational limitations currently faced by practitioners with massive datasets. In particular, a data subsampling or landmark selection process is often employed to construct a kernel based on partial information, followed by an approximate spectral analysis termed the Nystrom extension. We provide a quantitative framework to analyse this procedure, and use it to demonstrate algorithmic performance bounds on a range of practical approaches designed to optimize the landmark selection process. We compare the practical implications of these bounds by way of real-world examples drawn from the field of computer vision, whereby low-dimensional manifold structure is shown to emerge from high dimensional video data streams. PMID- 19805448 TI - Cherry-picking for complex data: robust structure discovery. AB - Complex data often arise as a superposition of data generated from several simpler models. The traditional strategy for such cases is to use mixture modelling, but it can be problematic, especially in higher dimensions. This paper considers an alternative approach, emphasizing data exploration and robustness to model misspecification. The strategy is applied to problems in regression, cluster analysis and multidimensional scaling. The approach is illustrated through simulation and the analysis of several datasets. PMID- 19805449 TI - Statistical inference for exploratory data analysis and model diagnostics. AB - We propose to furnish visual statistical methods with an inferential framework and protocol, modelled on confirmatory statistical testing. In this framework, plots take on the role of test statistics, and human cognition the role of statistical tests. Statistical significance of 'discoveries' is measured by having the human viewer compare the plot of the real dataset with collections of plots of simulated datasets. A simple but rigorous protocol that provides inferential validity is modelled after the 'lineup' popular from criminal legal procedures. Another protocol modelled after the 'Rorschach' inkblot test, well known from (pop-)psychology, will help analysts acclimatize to random variability before being exposed to the plot of the real data. The proposed protocols will be useful for exploratory data analysis, with reference datasets simulated by using a null assumption that structure is absent. The framework is also useful for model diagnostics in which case reference datasets are simulated from the model in question. This latter point follows up on previous proposals. Adopting the protocols will mean an adjustment in working procedures for data analysts, adding more rigour, and teachers might find that incorporating these protocols into the curriculum improves their students' statistical thinking. PMID- 19805450 TI - Sufficient dimension reduction and prediction in regression. AB - Dimension reduction for regression is a prominent issue today because technological advances now allow scientists to routinely formulate regressions in which the number of predictors is considerably larger than in the past. While several methods have been proposed to deal with such regressions, principal components (PCs) still seem to be the most widely used across the applied sciences. We give a broad overview of ideas underlying a particular class of methods for dimension reduction that includes PCs, along with an introduction to the corresponding methodology. New methods are proposed for prediction in regressions with many predictors. PMID- 19805451 TI - Identifying graph clusters using variational inference and links to covariance parametrization. AB - Finding clusters of well-connected nodes in a graph is a problem common to many domains, including social networks, the Internet and bioinformatics. From a computational viewpoint, finding these clusters or graph communities is a difficult problem. We use a clique matrix decomposition based on a statistical description that encourages clusters to be well connected and few in number. The formal intractability of inferring the clusters is addressed using a variational approximation inspired by mean-field theories in statistical mechanics. Clique matrices also play a natural role in parametrizing positive definite matrices under zero constraints on elements of the matrix. We show that clique matrices can parametrize all positive definite matrices restricted according to a decomposable graph and form a structured factor analysis approximation in the non decomposable case. Extensions to conjugate Bayesian covariance priors and more general non-Gaussian independence models are briefly discussed. PMID- 19805452 TI - Classification of sparse high-dimensional vectors. AB - We study the problem of classification of d-dimensional vectors into two classes (one of which is 'pure noise') based on a training sample of size m. The main specific feature is that the dimension d can be very large. We suppose that the difference between the distribution of the population and that of the noise is only in a shift, which is a sparse vector. For Gaussian noise, fixed sample size m, and dimension d that tends to infinity, we obtain the sharp classification boundary, i.e. the necessary and sufficient conditions for the possibility of successful classification. We propose classifiers attaining this boundary. We also give extensions of the result to the case where the sample size m depends on d and satisfies the condition (log m)/log d --> gamma, 0 6 months) are asymptomatic and do not generally require treatment. These children are, however, at increased risk for severe complications later in life, including advanced liver disease and liver cancer. On November 11, 2008, the Hepatitis B Foundation, a nonprofit research and disease advocacy organization, convened a panel of nationally recognized North American pediatric liver specialists to consider and recommend an approach for the screening, monitoring, initial management, and referral of children with chronic hepatitis B. The panel developed recommendations to provide guidance to practitioners on determining what additional tests to conduct, how often to monitor on the basis of test results, and when to refer to a pediatric liver specialist to build a partnership between the practitioner and liver specialist to enhance the success of management of children with this lifelong infection. PMID- 19805458 TI - Young driver education programs that build resilience have potential to reduce road crashes. AB - OBJECTIVE: The research aimed to explore associations between participation in 2 education programs for school-based learner drivers and subsequent road traffic offenses and crashes among a large cohort of newly licensed drivers. METHODS: DRIVE is a prospective cohort study of 20822 first-year drivers aged 17 to 24 in New South Wales (NSW), Australia. Participants completed a detailed questionnaire and consented to data linkage in 2003-2004. Questionnaire items included year of participation in 2 specific education programs: a 1-day workshop-only program focusing on driving risks ("driver-focused") and a whole-of-community program also including a 1-day workshop but also longer term follow-up activities and a broader focus on reducing risk-taking and building resilience ("resilience focused"). Survey data were subsequently linked to police-reported crash and offense data for 1996-2005. Poisson regression models that adjusted for multiple confounders were created to explore offenses and crashes as a driver (dichotomized as 0 vs >or=1) after program participation. RESULTS: Offenses did not differ between groups; however, whereas the driver-focused program was not associated with reduced crash risk, the resilience-focused program was associated with a 44% reduced relative risk for crash (0.56 [95% confidence interval: 0.34 0.93]). CONCLUSIONS: The large effect size observed and complementary findings from a comparable randomized, controlled trial in the United States suggest programs that focus more generally on reducing risks and building resilience have the potential to reduce crashes. A large, representative, randomized, controlled trial is urgently needed to confirm road safety benefits and ensure evidence based spending and practitioner recommendations in this field. PMID- 19805459 TI - Violence, abuse, and crime exposure in a national sample of children and youth. AB - OBJECTIVE: The objective of this research was to obtain national estimates of exposure to the full spectrum of the childhood violence, abuse, and crime victimizations relevant to both clinical practice and public-policy approaches to the problem. METHODS: The study was based on a cross-sectional national telephone survey that involved a target sample of 4549 children aged 0 to 17 years. RESULTS: A clear majority (60.6%) of the children and youth in this nationally representative sample had experienced at least 1 direct or witnessed victimization in the previous year. Almost half (46.3%) had experienced a physical assault in the study year, 1 in 4 (24.6%) had experienced a property offense, 1 in 10 (10.2%) had experienced a form of child maltreatment, 6.1% had experienced a sexual victimization, and more than 1 in 4 (25.3%) had been a witness to violence or experienced another form of indirect victimization in the year, including 9.8% who had witnessed an intrafamily assault. One in 10 (10.2%) had experienced a victimization-related injury. More than one third (38.7%) had been exposed to 2 or more direct victimizations, 10.9% had 5 or more, and 2.4% had 10 or more during the study year. CONCLUSIONS: The scope and diversity of child exposure to victimization is not well recognized. Clinicians and researchers need to inquire about a larger spectrum of victimization types to identify multiply victimized children and tailor prevention and interventions to the full range of threats that children face. PMID- 19805460 TI - Prevalence of parent-reported diagnosis of autism spectrum disorder among children in the US, 2007. AB - OBJECTIVES: The reported increasing prevalence of autism spectrum disorder (ASD) and attendant health and family impact make monitoring of ASD prevalence a public health priority. METHODS: The prevalence of parent-reported diagnosis of ASD among US children aged 3 to 17 years was estimated from the 2007 National Survey of Children's Health (sample size: 78037). A child was considered to have ASD if a parent/guardian reported that a doctor or other health care provider had ever said that the child had ASD and that the child currently had the condition. The point-prevalence for ASD was calculated for those children meeting both criteria. We examined sociodemographic factors associated with current ASD and with a past (but not current) ASD diagnosis. The health care experiences for children in both ASD groups were explored. RESULTS: The weighted current ASD point-prevalence was 110 per 10,000. We estimate that 673,000 US children have ASD. Odds of having ASD were 4 times as large for boys than girls. Non-Hispanic (NH) black and multiracial children had lower odds of ASD than NH white children. Nearly 40% of those ever diagnosed with ASD did not currently have the condition; NH black children were more likely than NH white children to not have current ASD. Children in both ASD groups were less likely than children without ASD to receive care within a medical home. CONCLUSIONS: The observed point-prevalence is higher than previous US estimates. More inclusive survey questions, increased population awareness, and improved screening and identification by providers may partly explain this finding. PMID- 19805462 TI - Ethnic group and medical care. What about doctor factors? PMID- 19805463 TI - Contraception for women. First consultation for the pill. PMID- 19805464 TI - Interpreting sickness absence rates of NHS employees. PMID- 19805467 TI - Equity and NHS reforms. Waiting times are poor marker for equity. PMID- 19805471 TI - Behcet's syndrome. PMID- 19805472 TI - Meeting the challenge of population ageing. PMID- 19805475 TI - Answering the burning question of how transient receptor potential vanilloid-1 channel antagonists cause unwanted hyperthermia. AB - The treatment of chronic pain with new therapies that lack the side effects of existing analgesics is one of medicine's great unmet needs. Toward this goal, antagonists of the transient receptor potential vanilloid-1 (TRPV1) channel have shown some promise. However, the development of these compounds has been hindered by an unpleasant on-target hyperthermic side effect. With compelling evidence, the accompanying critical review by Romanovsky et al. (p. 228) regarding TRPV1 takes a position on the sites of action of TRPV1 agonists and antagonists on the thermoregulatory system that controls this side effect. From this comes insight on potential ways to overcome the unwanted hyperthermic action of TRPV1 antagonists. PMID- 19805477 TI - Pituitary adenylate cyclase-activating polypeptide and its receptors: 20 years after the discovery. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a 38-amino acid C terminally alpha-amidated peptide that was first isolated 20 years ago from an ovine hypothalamic extract on the basis of its ability to stimulate cAMP formation in anterior pituitary cells (Miyata et al., 1989. PACAP belongs to the vasoactive intestinal polypeptide (VIP)-secretin-growth hormone-releasing hormone glucagon superfamily. The sequence of PACAP has been remarkably well conserved during evolution from protochordates to mammals, suggesting that PACAP is involved in the regulation of important biological functions. PACAP is widely distributed in the brain and peripheral organs, notably in the endocrine pancreas, gonads, respiratory and urogenital tracts. Characterization of the PACAP precursor has revealed the existence of a PACAP-related peptide, the activity of which remains unknown. Two types of PACAP binding sites have been characterized: type I binding sites exhibit a high affinity for PACAP and a much lower affinity for VIP, whereas type II binding sites have similar affinity for PACAP and VIP. Molecular cloning of PACAP receptors has shown the existence of three distinct receptor subtypes: the PACAP-specific PAC1-R, which is coupled to several transduction systems, and the PACAP/VIP-indifferent VPAC1-R and VPAC2-R, which are primarily coupled to adenylyl cyclase. PAC1-Rs are particularly abundant in the brain, the pituitary and the adrenal gland, whereas VPAC receptors are expressed mainly in lung, liver, and testis. The development of transgenic animal models and specific PACAP receptor ligands has strongly contributed to deciphering the various actions of PACAP. Consistent with the wide distribution of PACAP and its receptors, the peptide has now been shown to exert a large array of pharmacological effects and biological functions. The present report reviews the current knowledge concerning the pleiotropic actions of PACAP and discusses its possible use for future therapeutic applications. PMID- 19805478 TI - Therapeutic targeting of nuclear protein import in pathological cell conditions. AB - Proteins enter the nucleus through the nuclear pore complex. Once in the nucleus, some proteins, such as transcriptional regulators, can turn genes on or off, and change the composition of the cell and its function to meet the demands of its environment. This process of protein import into the nucleus is highly controlled and regulated by the expression or function of single cargoes, transport receptors, or the transport channels themselves. Thus, these components of the import process have an impact on transport capacity, which subsequently affects gene expression, signal transduction, and cell growth and development. With such a key position in the process of cell growth, it is reasonable to hypothesize that alterations in nuclear protein transport may play an important role in pathological cell conditions that have abnormal cell growth as a central feature. Indeed, there are now sufficient data to demonstrate that alterations in nuclear protein transport participate in alterations in cell proliferation and hypertrophy. Further study is needed to provide definitive proof that changes in nuclear protein import directly participate in the pathogenesis of diseases such as hypertension, atherosclerosis, cancer, viral infection, and diabetes. However, the data to date have, on select occasions, led to a clear association of alterations in nuclear transport with disease states. Furthermore,this research has led to the important identification of new targets within the process of nuclear protein import that hold therapeutic promise to inhibit viral replication, to improve drug delivery during cancer therapy, and, in general, to modify cell growth and viability during disease conditions. PMID- 19805479 TI - Regulation of skeletal muscle physiology and metabolism by peroxisome proliferator-activated receptor delta. AB - Agonists directed against the alpha and gamma isoforms of the peroxisome proliferator-activated receptors (PPARs) have become important for the respective treatment of hypertriglyceridemia and insulin resistance associated with metabolic disease. PPARdelta is the least well characterized of the three PPAR isoforms. Skeletal muscle insulin resistance is a primary risk factor for the development of type 2 diabetes. There is increasing evidence that PPARdelta is an important regulator of skeletal muscle metabolism, in particular, muscle lipid oxidation, highlighting the potential utility of this isoform as a drug target. In addition, PPARdelta seems to be a key regulator of skeletal muscle fiber type and a possible mediator of the adaptations noted in skeletal muscle in response to exercise. In this review we summarize the current status regarding the regulation, and the metabolic effects, of PPARdelta in skeletal muscle. PMID- 19805476 TI - Inflammatory stress and idiosyncratic hepatotoxicity: hints from animal models. AB - Adverse drug reactions (ADRs) present a serious human health problem. They are major contributors to hospitalization and mortality throughout the world (Lazarou et al., 1998; Pirmohamed et al., 2004). A small fraction (less than 5%) of ADRs can be classified as "idiosyncratic." Idiosyncratic ADRs (IADRs) are caused by drugs with diverse pharmacological effects and occur at various times during drug therapy. Although IADRs affect a number of organs, liver toxicity occurs frequently and is the primary focus of this review. Because of the inconsistency of clinical data and the lack of experimental animal models, how IADRs arise is largely undefined. Generation of toxic drug metabolites and induction of specific immunity are frequently cited as causes of IADRs, but definitive evidence supporting either mechanism is lacking for most drugs. Among the more recent hypotheses for causation of IADRs is that inflammatory stress induced by exogenous or endogenous inflammagens is a susceptibility factor. In this review, we give a brief overview of idiosyncratic hepatotoxicity and the inflammatory response induced by bacterial lipopolysaccharide. We discuss the inflammatory stress hypothesis and use as examples two drugs that have caused IADRs in human patients: ranitidine and diclofenac. The review focuses on experimental animal models that support the inflammatory stress hypothesis and on the mechanisms of hepatotoxic response in these models. The need for design of epidemiological studies and the potential for implementation of inflammation interaction studies in preclinical toxicity screening are also discussed briefly. PMID- 19805480 TI - Glucocorticoid receptor interacting protein-1 restores glucocorticoid responsiveness in steroid-resistant airway structural cells. AB - Glucocorticoid (GC) insensitivity represents a profound challenge in managing patients with asthma. The mutual inhibition of transcriptional activity between GC receptor (GR) and other regulators is one of the mechanisms contributing to GC resistance in asthma. We recently reported that interferon regulatory factor (IRF)-1 is a novel transcription factor that promotes GC insensitivity in human airway smooth muscle (ASM) cells by interfering with GR signaling (Tliba et al., Am J Respir Cell Mol Biol 2008;38:463-472). Here, we sought to determine whether the inhibition of GR function by IRF-1 involves its interaction with the transcriptional co-regulator GR-interacting protein 1 (GRIP-1), a known GR transcriptional co-activator. We here found that siRNA-mediated GRIP-1 depletion attenuated IRF-1-dependent transcription of the luciferase reporter construct and the mRNA expression of an IRF-1-dependent gene, CD38. In parallel experiments, GRIP-1 silencing significantly reduced GR-mediated transactivation activities. Co immunoprecipitation and GST pull-down assays showed that GRIP-1, through its repression domain, physically interacts with IRF-1 identifying GRIP-1 as a bona fide transcriptional co-activator for IRF-1. Interestingly, the previously reported inhibition of GR-mediated transactivation activities by either TNF-alpha and IFN-gamma treatment or IRF-1 overexpression was fully reversed by increasing cellular levels of GRIP-1. Together, these data suggest that the cellular accumulation of IRF-1 may represent a potential molecular mechanism mediating altered cellular response to GC through the depletion of GRIP-1 from the GR transcriptional regulatory complexes. PMID- 19805481 TI - Enhancement of Th1 lung immunity induced by recombinant Mycobacterium bovis Bacillus Calmette-Guerin attenuates airway allergic disease. AB - Mycobacterium bovis Bacillus Calmette-Guerin (BCG) has been shown to down regulate experimental allergic asthma, a finding that reinforced the hygiene hypothesis. We have previously found that recombinant BCG (rBCG) strain that express the genetically detoxified S1 subunit of pertussis toxin (rBCG-S1PT) exerts an adjuvant effect that enhances Th1 responses against BCG proteins. Here we investigated the effect of this rBCG-S1PT on the classical ovalbumin-induced mouse model of allergic lung disease. We found that rBCG-S1PT was more effective than wild-type BCG in preventing Th2-mediated allergic immune responses. The inhibition of allergic lung disease was not associated with increased concentration of suppressive cytokines or with an increased number of pulmonary regulatory T cells but was positively correlated with the increase in IFN-gamma producing T cells and T-bet expression in the lung. In addition, an IL-12 dependent mechanism appeared to be important to the inhibition of lung allergic disease. The inhibition of allergic inflammation was found to be restricted to the lung because when allergen challenge was given by the intraperitoneal route, rBCG-S1PT administration failed to inhibit peritoneal allergic inflammation and type 2 cytokine production. Our work offers a nonclassical interpretation for the hygiene hypothesis indicating that attenuation of lung allergy by rBCG could be due to the enhancement of local lung Th1 immunity induced by rBCG-S1PT. Moreover, it highlights the possible use of rBCG strains as multipurpose immunomodulators by inducing specific immunity against microbial products while protecting against allergic asthma. PMID- 19805482 TI - Adenovirus vectors block human immunodeficiency virus-1 replication in human alveolar macrophages by inhibition of the long terminal repeat. AB - Heterologous viruses may transactivate or suppress human immunodeficiency virus (HIV)-1 replication. An adenovirus type 5 gene transfer vector (Ad5) HIV-1 vaccine was recently evaluated in a clinical trial, without efficacy. In this context, it is relevant to ask what effect Ad vectors have on HIV-1 replication, particularly in cells that are part of the innate immune system. Infection of HIV 1-infected human alveolar macrophages (AMs) obtained from HIV-1(+) individuals with an Ad vector containing no transgene (AdNull) resulted in dose-responsive inhibition of endogenous HIV-1 replication. HIV-1 replication in normal AMs infected with HIV-1 in vitro was inhibited by AdNull with a similar dose response. Ad reduced AM HIV-1 replication up to 14 days after HIV-1 infection. Fully HIV-1-infected AMs were treated with 3'-azido-3'-deoxythymidine, after which Ad infection still inhibited HIV-1 replication, suggesting a postentry step was affected. Substantial HIV-1 DNA was still produced after Ad infection, as quantified by TaqMan real-time PCR, suggesting that the replication block occurred after reverse transcription. AdNull blocked HIV-1 long terminal repeat (LTR) transcription, as assessed by an vesicular stomatitis virus G protein pseudotyped HIV-1 LTR luciferase construct. The formation of HIV-1 DNA integrated into the host chromosome was not inhibited by Ad, as quantified by a two-step TaqMan real-time PCR assay, implying a postintegration block to HIV-1 replication. These data indicate that Ad vectors are inhibitory to HIV-1 replication in human AMs based, in part, on their ability to inhibit LTR-driven transcription. PMID- 19805483 TI - Both hematopoietic-derived and non-hematopoietic-derived {beta}-arrestin-2 regulates murine allergic airway disease. AB - Allergic asthma, a major cause of morbidity and leading cause of hospitalizations, is an inflammatory disease orchestrated by T helper cells and characterized by the lung migration of eosinophils, which are important asthma effector cells. Lung migration of inflammatory cells requires, among other events, the chemokine receptor transduction of lung-produced inflammatory chemokines. Despite the widespread prevalence of this disease, the molecular mechanisms regulating chemokine production and receptor regulation in asthma are poorly understood. Previous work from our laboratory demonstrated that beta arrestin-2 positively regulates the development of allergic airway disease in a mouse model, partly through positive regulation of T-lymphocyte chemotaxis to the lung. However, beta-arrestin-2 is expressed in many cell types, including other hematopoietic cells and lung structural cells, which are involved in the development and manifestation of allergic airway disease. To determine the cell types required for beta-arrestin-2-dependent allergic inflammation, we generated bone marrow chimera mice. Using the ovalbumin murine model of allergic airway disease, we show that eosinophilic and lymphocytic inflammation is restored in chimeric mice, with expression of beta-arrestin-2 exclusively on hematopoietic derived cell types. In contrast, airway hyperresponsiveness is dependent on the expression of beta-arrestin-2 in structural cells. Our data demonstrate that the expression of beta-arrestin-2 in at least two divergent cell types contributes to the pathogenesis of allergic airway disease. PMID- 19805485 TI - A multiphase model for tissue construct growth in a perfusion bioreactor. AB - The growth of a cell population within a rigid porous scaffold in a perfusion bioreactor is studied, using a three-phase continuum model of the type presented by Lemon et al. (2006, Multiphase modelling of tissue growth using the theory of mixtures. J. Math. Biol., 52, 571-594) to represent the cell population (and attendant extracellular matrix), culture medium and porous scaffold. The bioreactor system is modelled as a 2D channel containing the cell-seeded rigid porous scaffold (tissue construct) which is perfused with culture medium. The study concentrates on (i) the cell-cell and cell-scaffold interactions and (ii) the impact of mechanotransduction mechanisms on construct composition. A numerical and analytical analysis of the model equations is presented and, depending upon the relative importance of cell aggregation and repulsion, markedly different cell movement is revealed. Additionally, mechanotransduction effects due to cell density, pressure and shear stress-mediated tissue growth are shown to generate qualitative differences in the composition of the resulting construct. The results of our simulations indicate that this model formulation (in conjunction with appropriate experimental data) has the potential to provide a means of identifying the dominant regulatory stimuli in a cell population. PMID- 19805486 TI - Recognition of facial, auditory, and bodily emotions in older adults. AB - Understanding older adults' social functioning difficulties requires insight into their recognition of emotion processing in voices and bodies, not just faces, the focus of most prior research. We examined 60 young and 61 older adults' recognition of basic emotions in facial, vocal, and bodily expressions, and when matching faces and bodies to voices, using 120 emotion items. Older adults were worse than young adults in 17 of 30 comparisons, with consistent difficulties in recognizing both positive (happy) and negative (angry and sad) vocal and bodily expressions. Nearly three quarters of older adults functioned at a level similar to the lowest one fourth of young adults, suggesting that age-related changes are common. In addition, we found that older adults' difficulty in matching emotions was not explained by difficulty on the component sources (i.e., faces or voices on their own), suggesting an additional problem of integration. PMID- 19805484 TI - Nuclear erythroid 2 p45-related factor 2 inhibits the maturation of murine dendritic cells by ragweed extract. AB - Oxidative stress plays an important role in immune regulation and dendritic cell (DC) maturation. Recent studies indicate that allergens, including ragweed extract (RWE), possess prooxidant activities, but how RWE interacts with DCs is not well understood. Nuclear erythroid 2 p45-related factor 2 (Nrf2) is a key transcription factor that regulates constitutive and coordinated induction of a battery of antioxidant genes. We hypothesized that RWE would activate DCs and that this response would be augmented in the absence of Nrf2. We generated bone marrow-derived DCs (BM-DCs) and isolated lung DCs from Nrf2(+/+) and Nrf2(-/-) mice and studied the effects of RWE on DCs in vitro. Under resting conditions, Nrf2(-/-) BM-DCs exhibited constitutively greater levels of inflammatory cytokines and costimulatory molecules than Nrf2(+/+) BM-DCs. Exposure to RWE impaired endocytic activity, significantly induced oxidative stress, and enhanced the expression of CD80, CD86, and MHCII in Nrf2(-/-) BM-DCs when compared with Nrf2(+/+) BM-DC, in association with reduced expression of Nrf2-regulated antioxidant genes. RWE significantly induced the secretion of inflammatory cytokines IL-6 and TNF-alpha in BM-DCs and lung DCs from Nrf2(-/-) mice than Nrf2(+/+) mice and significantly inhibited the secretion of IL-12 in Nrf2(+/+) BM DCs and IL-18 in Nrf2(+/+) and Nrf2(-/-) BM-DCs. The stimulatory effects of RWE on DC activation were inhibited to varying degrees by the antioxidant N-acetyl cysteine. Our findings indicate that a defect in Nrf2-mediated signaling mechanisms alters the response of DCs to a common environmental allergen, which may contribute to the susceptibility to allergic diseases. PMID- 19805487 TI - Comparison of patient and caregiver reports of patient activity participation and its relationship to mental health in patients with Alzheimer's disease. AB - The relationship between engagement in pleasant activities as rated by the patient and as rated by the caregiver from the patient's perspective was examined using structural equation modeling in a sample of patients (N = 277) diagnosed with mild to moderate Alzheimer's disease. The two activity participation ratings were only moderately related to one another. Furthermore, depression was the only significant predictor of the patient-rated activity participation, whereas severity of depression, degree of personality change, level of dependence, and cognition were all significant predictors of caregiver-rated activity participation. These findings suggest that caregivers consider a wider range of variables when evaluating the patient's engagement in activities than does the patient. Predictors of patient-rated activity participation did not differ as a function of age or cognition. PMID- 19805488 TI - Cessation of alcohol drinking, tobacco smoking and the reversal of head and neck cancer risk. AB - BACKGROUND: Quitting tobacco or alcohol use has been reported to reduce the head and neck cancer risk in previous studies. However, it is unclear how many years must pass following cessation of these habits before the risk is reduced, and whether the risk ultimately declines to the level of never smokers or never drinkers. METHODS: We pooled individual-level data from case-control studies in the International Head and Neck Cancer Epidemiology Consortium. Data were available from 13 studies on drinking cessation (9167 cases and 12 593 controls), and from 17 studies on smoking cessation (12 040 cases and 16 884 controls). We estimated the effect of quitting smoking and drinking on the risk of head and neck cancer and its subsites, by calculating odds ratios (ORs) using logistic regression models. RESULTS: Quitting tobacco smoking for 1-4 years resulted in a head and neck cancer risk reduction [OR 0.70, confidence interval (CI) 0.61-0.81 compared with current smoking], with the risk reduction due to smoking cessation after > or =20 years (OR 0.23, CI 0.18-0.31), reaching the level of never smokers. For alcohol use, a beneficial effect on the risk of head and neck cancer was only observed after > or =20 years of quitting (OR 0.60, CI 0.40-0.89 compared with current drinking), reaching the level of never drinkers. CONCLUSIONS: Our results support that cessation of tobacco smoking and cessation of alcohol drinking protect against the development of head and neck cancer. PMID- 19805490 TI - Commentary: a Darwin family concern. PMID- 19805491 TI - Proximal dentatothalamocortical tract involvement in posterior fossa syndrome. AB - Posterior fossa syndrome is characterized by cerebellar dysfunction, oromotor/oculomotor apraxia, emotional lability and mutism in patients after infratentorial injury. The underlying neuroanatomical substrates of posterior fossa syndrome are unknown, but dentatothalamocortical tracts have been implicated. We used pre- and postoperative neuroimaging to investigate proximal dentatothalamocortical tract involvement in childhood embryonal brain tumour patients who developed posterior fossa syndrome following tumour resection. Diagnostic imaging from a cohort of 26 paediatric patients previously operated on for an embryonal brain tumour (13 patients prospectively diagnosed with posterior fossa syndrome, and 13 non-affected patients) were evaluated. Preoperative magnetic resonance imaging was used to define relevant tumour features, including two potentially predictive measures. Postoperative magnetic resonance and diffusion tensor imaging were used to characterize operative injury and tract based differences in anisotropy of water diffusion. In patients who developed posterior fossa syndrome, initial tumour resided higher in the 4th ventricle (P = 0.035). Postoperative magnetic resonance signal abnormalities within the superior cerebellar peduncles and midbrain were observed more often in patients with posterior fossa syndrome (P = 0.030 and 0.003, respectively). The fractional anisotropy of water was lower in the bilateral superior cerebellar peduncles, in the bilateral fornices, white matter region proximate to the right angular gyrus (Tailerach coordinates 35, -71, 19) and white matter region proximate to the left superior frontal gyrus (Tailerach coordinates -24, 57, 20). Our findings suggest that multiple bilateral injuries to the proximal dentatothalamocortical pathways may predispose the development of posterior fossa syndrome, that functional disruption of the white matter bundles containing efferent axons within the superior cerebellar peduncles is a critical underlying pathophysiological component of posterior fossa syndrome, and that decreased fractional anisotropy in the fornices and cerebral cortex may be related to the abnormal neurobehavioural symptoms of posterior fossa syndrome. PMID- 19805492 TI - Semantic dementia: demography, familial factors and survival in a consecutive series of 100 cases. AB - A great deal has been written about cognitive aspects of semantic dementia but little is known about the demography or prognosis. We describe these features in a consecutive series of 100 patients seen over a 17-year period; all cases were assessed and followed up in a specialist clinic. The mean age at diagnosis was 64.2 (+/-7.1) range 40-79 years, but 46 presented after age 65 and 7 after 75; a higher proportion than the existing literature might predict. Fifteen had a first degree relative with dementia, but in seven this was almost certainly unrelated. Only two had relatives with young-onset dementia. There were no families with more than two affected members. The familial rate was estimated at between 2% and 7% (95% confidence interval 0-12%). Kaplan-Meier analyses indicated a 50% survival of 12.8 years (95% confidence interval 11.9-13.7); a more benign course than suggested by neuropathologically based studies. We were unable to identify any factors influencing survival. Of the 100, 34 have died, with pathological confirmation in 24; 18 had frontotemporal lobar degeneration with ubiquitin positive inclusions (13 of 13 confirmed TAR DNA binding protein-43 positive), and 3 had classic tau-positive Pick bodies and 3 had Alzheimer's pathology. The age at diagnosis or death across the pathological subgroups was equivalent. Although semantic dementia has a strong statistical association with ubiquitin-positive pathology, it does not have the signature of familial frontotemporal lobar degeneration with ubiquitin-positive inclusions, notably the presence of intranuclear lentiform TAR DNA binding protein-43 inclusions. The age of onset is older than predicted and the course more slowly progressive than suggested by earlier studies of small groups of subjects. PMID- 19805493 TI - Microglial CB2 cannabinoid receptors are neuroprotective in Huntington's disease excitotoxicity. AB - Cannabinoid-derived drugs are promising agents for the development of novel neuroprotective strategies. Activation of neuronal CB(1) cannabinoid receptors attenuates excitotoxic glutamatergic neurotransmission, triggers prosurvival signalling pathways and palliates motor symptoms in animal models of neurodegenerative disorders. However, in Huntington's disease there is a very early downregulation of CB(1) receptors in striatal neurons that, together with the undesirable psychoactive effects triggered by CB(1) receptor activation, foster the search for alternative pharmacological treatments. Here, we show that CB(2) cannabinoid receptor expression increases in striatal microglia of Huntington's disease transgenic mouse models and patients. Genetic ablation of CB(2) receptors in R6/2 mice, that express human mutant huntingtin exon 1, enhanced microglial activation, aggravated disease symptomatology and reduced mice lifespan. Likewise, induction of striatal excitotoxicity in CB(2) receptor deficient mice by quinolinic acid administration exacerbated brain oedema, microglial activation, proinflammatory-mediator state and medium-sized spiny neuron degeneration. Moreover, administration of CB(2) receptor-selective agonists to wild-type mice subjected to excitotoxicity reduced neuroinflammation, brain oedema, striatal neuronal loss and motor symptoms. Studies on ganciclovir induced depletion of astroglial proliferation in transgenic mice expressing thymidine kinase under the control of the glial fibrillary acidic protein promoter excluded the participation of proliferating astroglia in CB(2) receptor mediated actions. These findings support a pivotal role for CB(2) receptors in attenuating microglial activation and preventing neurodegeneration that may pave the way to new therapeutic strategies for neuroprotection in Huntington's disease as well as in other neurodegenerative disorders with a significant excitotoxic component. PMID- 19805494 TI - Relating visual to verbal semantic knowledge: the evaluation of object recognition in prosopagnosia. AB - Assessment of face specificity in prosopagnosia is hampered by difficulty in gauging pre-morbid expertise for non-face object categories, for which humans vary widely in interest and experience. In this study, we examined the correlation between visual and verbal semantic knowledge for cars to determine if visual recognition accuracy could be predicted from verbal semantic scores. We had 33 healthy subjects and six prosopagnosic patients first rated their own knowledge of cars. They were then given a test of verbal semantic knowledge that presented them with the names of car models, to which they were to match the manufacturer. Lastly, they were given a test of visual recognition, presenting them with images of cars to which they were to provide information at three levels of specificity: model, manufacturer and decade of make. In controls, while self-ratings were only moderately correlated with either visual recognition or verbal semantic knowledge, verbal semantic knowledge was highly correlated with visual recognition, particularly for more specific levels of information. Item concordance showed that less-expert subjects were more likely to provide the most specific information (model name) for the image when they could also match the manufacturer to its name. Prosopagnosic subjects showed reduced visual recognition of cars after adjusting for verbal semantic scores. We conclude that visual recognition is highly correlated with verbal semantic knowledge, that formal measures of verbal semantic knowledge are a more accurate gauge of expertise than self-ratings, and that verbal semantic knowledge can be used to adjust tests of visual recognition for pre-morbid expertise in prosopagnosia. PMID- 19805498 TI - Clinical images: Spontaneous rectus sheath hematoma. PMID- 19805497 TI - Effect of ambient air pollution on the incidence of appendicitis. AB - BACKGROUND: The pathogenesis of appendicitis is unclear. We evaluated whether exposure to air pollution was associated with an increased incidence of appendicitis. METHODS: We identified 5191 adults who had been admitted to hospital with appendicitis between Apr. 1, 1999, and Dec. 31, 2006. The air pollutants studied were ozone, nitrogen dioxide, sulfur dioxide, carbon monoxide, and suspended particulate matter of less than 10 micro and less than 2.5 micro in diameter. We estimated the odds of appendicitis relative to short-term increases in concentrations of selected pollutants, alone and in combination, after controlling for temperature and relative humidity as well as the effects of age, sex and season. RESULTS: An increase in the interquartile range of the 5-day average of ozone was associated with appendicitis (odds ratio [OR] 1.14, 95% confidence interval [CI] 1.03-1.25). In summer (July-August), the effects were most pronounced for ozone (OR 1.32, 95% CI 1.10-1.57), sulfur dioxide (OR 1.30, 95% CI 1.03-1.63), nitrogen dioxide (OR 1.76, 95% CI 1.20-2.58), carbon monoxide (OR 1.35, 95% CI 1.01-1.80) and particulate matter less than 10 micro in diameter (OR 1.20, 95% CI 1.05-1.38). We observed a significant effect of the air pollutants in the summer months among men but not among women (e.g., OR for increase in the 5-day average of nitrogen dioxide 2.05, 95% CI 1.21-3.47, among men and 1.48, 95% CI 0.85-2.59, among women). The double-pollutant model of exposure to ozone and nitrogen dioxide in the summer months was associated with attenuation of the effects of ozone (OR 1.22, 95% CI 1.01-1.48) and nitrogen dioxide (OR 1.48, 95% CI 0.97-2.24). INTERPRETATION: Our findings suggest that some cases of appendicitis may be triggered by short-term exposure to air pollution. If these findings are confirmed, measures to improve air quality may help to decrease rates of appendicitis. PMID- 19805499 TI - Keeping children safe: rethinking how we design our surroundings. PMID- 19805500 TI - China's counterfeit medicine trade booming. PMID- 19805501 TI - Stark contrasts. PMID- 19805502 TI - Shift toward capitation in Ontario. PMID- 19805489 TI - Causes of death in HIV-infected women: persistent role of AIDS. The 'Mortalite 2000 & 2005' Surveys (ANRS EN19). AB - BACKGROUND: Little is known about the causes of death in human immunodeficiency virus (HIV)-infected women in the era of combination antiretroviral therapy (ART). METHODS: In the French nationwide Mortalite 2000 and 2005 surveys, physicians reported causes of deaths in HIV-infected adults in 2000 and 2005, using a standardized questionnaire. We used multivariate logistic regression models to study the association between gender and AIDS-defining causes of death, adjusting for other characteristics. RESULTS: Of the 1013 HIV-infected adults who died in 2005, 247 (24%) were women. Half of women were infected through heterosexual contacts, compared with 25% men. In 2005, the proportion of AIDS defining causes of death was higher in women than in men (43 vs 34%; P = 0.01), whereas it had been the same in 2000 (47% in women and men). In 2005, women died less frequently than men from respiratory malignancies (lung, ear/nose/throat) and cardiovascular disease (9% of all causes of death in women compared with 16% in men; P = 0.004), and suicides or accidents (4 vs 9%; P = 0.02). Socio-economic precariousness, younger age, less alcohol and tobacco consumption and lack of prior ART explained the higher proportion of deaths from AIDS in women compared with men. CONCLUSIONS: The higher proportion of AIDS-related deaths in women is probably explained by two factors: (i) some HIV-infected women, especially migrants in poor socio-economic conditions, may not have access to optimal care; and (ii) a lower prevalence of risk factors for respiratory, cardiovascular and violent deaths means that the risk of dying from non-AIDS causes may be lower in women. PMID- 19805503 TI - Conflict emerges over value of handwashing. PMID- 19805504 TI - Stented within a stentless aortic valve. A simple surgical solution for the replacement of a stentless aortic bioprosthesis. AB - Stentless aortic bioprostheses are designed to offer better hemodynamics, less mechanical stress to the leaflets and as a result less degeneration. Although encouraging results are reported, little evidence has been published regarding reoperations of stentless valves. We are reporting a case of a structural valve dysfunction of an O'Brien-Angell stentless prosthesis, which could not be extracted during reoperation without damaging the aortic root. We are presenting a simple, quick and effective surgical solution, the surgical 'valve within a valve' technique for the avoidance of a redo complex root procedure. PMID- 19805505 TI - Isolated rib metastases from renal cell carcinoma. AB - Osseous metastases of renal cell carcinoma (RCC) are the second most frequent location after lung metastases. They rarely present as isolated location. When isolated, resection may offer five-year survival rates of 30-60%. The purpose of the current study is to focus on a particular subset, the isolated rib metastases (IRM). The files of six patients who underwent radical resection for IRM were reviewed. All had previous radical nephrectomy for clear-cell renal cancer. The mean age of these six men was 55.3 years. Preoperative evaluation included in all patients a conventional chest radiograph and thoracic computed tomography (CT) scanning. Chest wall resections were wide and curative. The mean disease-free interval (DFI) after renal cancer treatment was 25 months. There was no postoperative death. Two patients had synchronous disease. One of them developed two recurrences operated on by large resections. They survived for 77 and 81 months. The overall five and ten-year survival rates were respectively, 83 and 66.7%. IRM of RCC are rare and remain not well-known. Surgical wide resection is a safe and effective treatment. PMID- 19805506 TI - Influence of cannabis use trajectories, grade repetition and family background on the school-dropout rate at the age of 17 years in France. AB - BACKGROUND: Research has shown that cannabis use contributes to school dropout, but few studies have distinguished the age at onset of use from the age at progression to daily use neither their interaction with grade repetition. METHODS: This study is based on a French representative cross-sectional survey (N = 29,393 teenagers aged 17 years) and uses retrospective data. The influence of drug-use patterns <16 years of age on school-dropout rates (5.3%) are modelled with logistic regressions among boys and girls. RESULTS: The main factors associated with dropout were a low family socio-economic status, early grade repetition, single-parent families and daily tobacco smoking (ORa > or = 2.6). The link with the move to daily cannabis use was more evident when it occurred <14 years of age (ORa = 2.05 for boys and 3.41 for girls) rather than at > or =15 years (ORa = 1.45 for both sexes). The onset of cannabis use was not significant when occurring <14 years of age, but was linked to school attainment when occurring at age 15-16 years (ORa = 0.80 for boys and 0.64 for girls). Results are similar for alcohol use. Repeating a grade before beginning to use cannabis increased the dropout rates compared with the opposite sequence. Girls were more affected by early grade repetition and by early and daily cannabis use. CONCLUSION: Cannabis use is rarely a trigger for grade repetition but can have either damaging or positive effects on school attainment depending of the level of use. Positive social competence reflected by peer initiation should be investigated to understand this paradoxical effect. PMID- 19805507 TI - Soft drink consumption in adolescence: associations with food-related lifestyles and family rules in Belgium Flanders and the Veneto Region of Italy. AB - BACKGROUND: The number of studies among adolescents that focus on several lifestyle behaviours and family rules as determinant of soft drink consumption are limited. The aim of this study is to investigate the associations between daily soft drink consumption, food-related lifestyles and family rules in adolescence. METHODS: The data are part of the Health Behavior in School-aged Children (HBSC) cross-sectional survey. Adolescents between 11 and 16 years of age were included, resulting in a final sample of 14 407 adolescents representative of Belgium Flanders (N = 7904) and the Veneto Region of Italy (N = 6503). Binary logistic regression was used to test the association between soft drink consumption and food-related lifestyle (breakfast habits, family meals, snacking, meals in fast food restaurants and television viewing) and family rules (restriction and obligation rules) by region and gender. RESULTS: Each independent variable is significantly associated with daily soft drink consumption, despite some sub-groups exceptions. When we entered all the variables into the same statistical model, the positive association with daily soft drink consumption remained significant for frequent meals in fast food restaurants, television variables and low restriction rules. Breakfast during weekdays, evening meal with parents and obligation rules remained significant only in specific sub-groups and not the entire sample. Finally, the association with breakfast with parents and during the weekend disappeared. CONCLUSION: These findings suggest that considering gender and cultural differences, involving parents and limiting adolescents' exposure to television would increase the effectiveness of interventions aimed to reduce soft drink consumption in adolescence. PMID- 19805508 TI - Pharmacological removal of human ether-a-go-go-related gene potassium channel inactivation by 3-nitro-N-(4-phenoxyphenyl) benzamide (ICA-105574). AB - Human ether-a-go-go-related gene (hERG) potassium channel activity helps shape the cardiac action potential and influences its duration. In this study, we report the discovery of 3-nitro-N-(4-phenoxyphenyl) benzamide (ICA-105574), a potent and efficacious hERG channel activator with a unique mechanism of action. In whole-cell patch-clamp studies of recombinant hERG channels, ICA-105574 steeply potentiated current amplitudes more than 10-fold with an EC(50) value of 0.5 +/- 0.1 microM and a Hill slope (n(H)) of 3.3 +/- 0.2. The effect on hERG channels was confirmed because the known hERG channel blockers, N-[4-[[1-[2-(6 methyl-2-pyridinyl)ethyl]-4-piperidinyl]carbonyl]phenyl]methanesulfonamide, 2HCl (E-4031) and BeKm-1, potently blocked the stimulatory effects of ICA-105574. The primary mechanism by which ICA-105574 potentiates hERG channel activity is by removing hERG channel inactivation, because ICA-105574 (2 microM) shifts the midpoint of the voltage-dependence of inactivation by >180 mV from -86 to +96 mV. In addition to the effects on inactivation, greater concentrations of ICA-105574 (3 microM) produced comparatively small hyperpolarizing shifts (up to 11 mV) in the voltage-dependence of channel activation and a 2-fold slowing of channel deactivation. In isolated guinea pig ventricular cardiac myocytes, ICA-105574 induced a concentration-dependent shortening of action potential duration (>70%, 3 microM) that could be prevented by preincubation with E-4031. In conclusion, we identified a novel agent that can prevent the inactivation of hERG potassium channels. This compound may provide a useful tool to further understand the mechanism by which hERG channels inactivate and affect cardiac function in addition to the role of hERG channels in other cell systems. PMID- 19805509 TI - Protein kinase C-mediated inhibition of recombinant T-type Cav3.2 channels by neurokinin 1 receptors. AB - The voltage-activated T-type calcium channel (Ca(V)3.2) and the G protein-coupled neurokinin 1 (NK1) receptor are expressed in peripheral tissues and in central neurons, in which they participate in diverse physiological processes, including neurogenic inflammation and nociception. In the present report, we demonstrate that recombinant Ca(V)3.2 channels are reversibly inhibited by NK1 receptors when both proteins are transiently coexpressed in human embryonic kidney 293 cells. We found that the voltage-dependent macroscopic properties of Ca(V)3.2 currents were unaffected during NK1 receptor-mediated inhibition. However, inhibition was attenuated in cells coexpressing either the dominant-negative Galpha(q) Q209L/D277N or the regulator of G protein signaling (RGS) proteins 2 (RGS2) and 3T (RGS3T), which are effective antagonists of Galpha(q/11). By contrast, inhibition was unaffected in cells coexpressing human rod transducin (Galpha(t)), which buffers Gbetagamma. Channel inhibition was blocked by 1-[6-[[17beta methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione (U73122) and bisindolylmaleimide I, selective inhibitors of phospholipase Cbeta and protein kinase C (PKC), respectively. Inhibition was occluded by application of the PKC activator phorbol-12-myristate-13-acetate. Altogether, these data indicate that NK1 receptors inhibit Ca(V)3.2 channels through a voltage-independent signaling pathway that involves Galpha(q/11), phospholipase Cbeta, and PKC. Our results provide novel evidence regarding the mechanisms underlying T-type calcium channel modulation by G protein-coupled receptors. Functional coupling between Ca(V)3.2 channels and NK1 receptors may be relevant in neurogenic inflammation, neuronal rhythmogenesis, nociception, and other physiological processes. PMID- 19805510 TI - The emerging role of HP1 in the DNA damage response. AB - Heterochromatin protein 1 (HP1) family members are versatile proteins involved in transcription, chromatin organization, and replication. Recent findings now have implicated HP1 proteins in the DNA damage response as well. Cell-biological approaches showed that reducing the levels of all three HP1 isoforms enhances DNA repair, possibly due to heterochromatin relaxation. Additionally, HP1 is phosphorylated in response to DNA damage, which was suggested to initiate the DNA damage response. These findings have led to the conclusion that heterochromatic proteins are inhibitory to repair and that their dissociation from heterochromatin may facilitate repair. In contrast with an inhibitory role, a more active role for HP1 in DNA repair also was proposed based on the finding that all HP1 isoforms are recruited to UV-induced lesions, oxidative lesions, and DNA breaks. The loss of HP1 renders nematodes highly sensitive to DNA damage, and mice lacking HP1beta suffer from genomic instability, suggesting that the loss of HP1 is not necessarily beneficial for repair. These findings raise the possibility that HP1 facilitates DNA repair by reorganizing chromatin, which may involve interactions between phosphorylated HP1 and other DNA damage response proteins. Taken together, these studies illustrate an emerging role of HP1 proteins in the response to genotoxic stress. PMID- 19805511 TI - Mechanism of Mpk1 mitogen-activated protein kinase binding to the Swi4 transcription factor and its regulation by a novel caffeine-induced phosphorylation. AB - The Mpk1 mitogen-activated protein kinase (MAPK) of the cell wall integrity signaling pathway uses a noncatalytic mechanism to activate the SBF (Swi4/Swi6) transcription factor. Active Mpk1 forms a complex with Swi4, the DNA-binding subunit of SBF, conferring the ability to bind DNA. Because SBF activation is independent of Mpk1 catalytic activity but requires Mpk1 to be in an active conformation, we sought to understand how Mpk1 interacts with Swi4. Mutational analysis revealed that binding and activation of Swi4 by Mpk1 requires an intact D-motif-binding site, a docking surface common to MAPKs that resides distal to the phosphorylation loop but does not require the substrate-binding site, revealing a novel mechanism for MAPK target regulation. Additionally, we found that Mpk1 binds near the autoinhibitory C terminus of Swi4, suggesting an activation mechanism in which Mpk1 substitutes for Swi6 in promoting Swi4 DNA binding. Finally, we show that caffeine is an atypical activator of cell wall integrity signaling, because it induces phosphorylation of the Mpk1 C-terminal extension at Ser423 and Ser428. These phosphorylations were dependent on the DNA damage checkpoint kinases, Mec1/Tel1 and Rad53. Phosphorylation of Ser423 specifically blocked SBF activation by preventing Mpk1 association with Swi4, revealing a novel mechanism for regulating MAPK target specificity. PMID- 19805512 TI - Alternative splicing modulates autoinhibition and SH3 accessibility in the Src kinase Fyn. AB - Src family kinases are central regulators of a large number of signaling pathways. To adapt to the idiosyncrasies of different cell types, these kinases may need a fine-tuning of their intrinsic molecular control mechanisms. Here, we describe on a molecular level how the Fyn kinase uses alternative splicing to adapt to different cellular environments. Using structural analysis, site directed mutagenesis, and functional analysis, we show how the inclusion of either exon 7A or 7B affects the autoinhibition of Fyn and how this changes the SH3-dependent interaction and tyrosine phosphorylation of Sam68, with functional consequences for the Sam68-regulated survival of epithelial cells. Our results illustrate a novel mechanism of evolution that may contribute to the complexity of Src kinase regulation. PMID- 19805513 TI - XPF-ERCC1 participates in the Fanconi anemia pathway of cross-link repair. AB - Interstrand cross-links (ICLs) prevent DNA strand separation and, therefore, transcription and replication, making them extremely cytotoxic. The precise mechanism by which ICLs are removed from mammalian genomes largely remains elusive. Genetic evidence implicates ATR, the Fanconi anemia proteins, proteins required for homologous recombination, translesion synthesis, and at least two endonucleases, MUS81-EME1 and XPF-ERCC1. ICLs cause replication-dependent DNA double-strand breaks (DSBs), and MUS81-EME1 facilitates DSB formation. The subsequent repair of these DSBs occurs via homologous recombination after the ICL is unhooked by XPF-ERCC1. Here, we examined the effect of the loss of either nuclease on FANCD2 monoubiquitination to determine if the nucleolytic processing of ICLs is required for the activation of the Fanconi anemia pathway. FANCD2 was monoubiquitinated in Mus81(-/-), Ercc1(-/-), and XPF-deficient human, mouse, and hamster cells exposed to cross-linking agents. However, the monoubiquitinated form of FANCD2 persisted longer in XPF-ERCC1-deficient cells than in wild-type cells. Moreover, the levels of chromatin-bound FANCD2 were dramatically reduced and the number of ICL-induced FANCD2 foci significantly lower in XPF-ERCC1 deficient cells. These data demonstrate that the unhooking of an ICL by XPF-ERCC1 is necessary for the stable localization of FANCD2 to the chromatin and subsequent homologous recombination-mediated DSB repair. PMID- 19805514 TI - Mammalian casein kinase 1alpha and its leishmanial ortholog regulate stability of IFNAR1 and type I interferon signaling. AB - Phosphorylation of the degron of the IFNAR1 chain of the type I interferon (IFN) receptor triggers ubiquitination and degradation of this receptor and, therefore, plays a crucial role in negative regulation of IFN-alpha/beta signaling. Besides the IFN-stimulated and Jak activity-dependent pathways, a basal ligand independent phosphorylation of IFNAR1 has been described and implicated in downregulating IFNAR1 in response to virus-induced endoplasmic reticulum (ER) stress. Here we report purification and characterization of casein kinase 1alpha (CK1alpha) as a bona fide major IFNAR1 kinase that confers basal turnover of IFNAR1 and cooperates with ER stress stimuli to mediate phosphorylation-dependent degradation of IFNAR1. Activity of CK1alpha was required for phosphorylation and downregulation of IFNAR1 in response to ER stress and viral infection. While many forms of CK1 were capable of phosphorylating IFNAR1 in vitro, human CK1alpha and L-CK1 produced by the protozoan Leishmania major were also capable of increasing IFNAR1 degron phosphorylation in cells. Expression of leishmania CK1 in mammalian cells stimulated the phosphorylation-dependent downregulation of IFNAR1 and attenuated its signaling. Infection of mammalian cells with L. major modestly decreased IFNAR1 levels and attenuated cellular responses to IFN-alpha in vitro. We propose a role for mammalian and parasite CK1 enzymes in regulating IFNAR1 stability and type I IFN signaling. PMID- 19805515 TI - Transcription factor Glis3, a novel critical player in the regulation of pancreatic beta-cell development and insulin gene expression. AB - In this study, we report that the Kruppel-like zinc finger transcription factor Gli-similar 3 (Glis3) is induced during the secondary transition of pancreatic development, a stage of cell lineage specification and extensive patterning, and that Glis3(zf/zf) mutant mice develop neonatal diabetes, evidenced by hyperglycemia and hypoinsulinemia. The Glis3(zf/zf) mutant mouse pancreas shows a dramatic loss of beta and delta cells, contrasting a smaller relative loss of alpha, PP, and epsilon cells. In addition, Glis3(zf/zf) mutant mice develop ductal cysts, while no significant changes were observed in acini. Gene expression profiling and immunofluorescent staining demonstrated that the expression of pancreatic hormones and several transcription factors important in endocrine cell development, including Ngn3, MafA, and Pdx1, were significantly decreased in the developing pancreata of Glis3(zf/zf) mutant mice. The population of pancreatic progenitors appears not to be greatly affected in Glis3(zf/zf) mutant mice; however, the number of neurogenin 3 (Ngn3)-positive endocrine cell progenitors is significantly reduced. Our study indicates that Glis3 plays a key role in cell lineage specification, particularly in the development of mature pancreatic beta cells. In addition, we provide evidence that Glis3 regulates insulin gene expression through two Glis-binding sites in its proximal promoter, indicating that Glis3 also regulates beta-cell function. PMID- 19805516 TI - Functional specificities of Brm and Brg-1 Swi/Snf ATPases in the feedback regulation of hepatic bile acid biosynthesis. AB - Bile acid homeostasis is critical in maintaining health and is primarily regulated by the nuclear receptors farnesoid X receptor (FXR) and small heterodimer partner (SHP). Bile acid-activated FXR indirectly inhibits expression of cholesterol 7alpha hydroxylase (CYP7A1), a key enzyme in conversion of cholesterol to bile acids, by induction of SHP. We recently demonstrated that SHP inhibits CYP7A1 transcription by recruiting chromatin-modifying cofactors, including Brm-Swi/Snf. Swi/Snf complexes contain either Brm or Brg-1 ATPases, and whether these subunits have distinct functions remains unclear. We have examined the role of these subunits in regulation of bile acid metabolism under physiological conditions by FXR and SHP. Brg-1 interacted with FXR and enhanced FXR-mediated transactivation of SHP, whereas Brm interacted with SHP and enhanced SHP-mediated repression of CYP7A1 and, interestingly, auto-repression of SHP. Chromatin immunoprecipitation and remodeling studies revealed that after treatment with FXR agonists, Brg-1 was recruited to the SHP promoter, resulting in transcriptionally active accessible chromatin, whereas Brm was recruited to both CYP7A1 and SHP promoters, resulting in inactive inaccessible chromatin. Our studies demonstrate that Brm and Brg-1 have distinct functions in the regulation of two key genes, CYP7A1 and SHP, within a single physiological pathway, feedback inhibition of bile acid biosynthesis, by differentially targeting SHP and FXR. PMID- 19805517 TI - Peroxisome proliferator-activated receptor beta/delta (PPARbeta/delta) but not PPARalpha serves as a plasma free fatty acid sensor in liver. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is an important transcription factor in liver that can be activated physiologically by fasting or pharmacologically by using high-affinity synthetic agonists. Here we initially set out to elucidate the similarities in gene induction between Wy14643 and fasting. Numerous genes were commonly regulated in liver between the two treatments, including many classical PPARalpha target genes, such as Aldh3a2 and Cpt2. Remarkably, several genes induced by Wy14643 were upregulated by fasting independently of PPARalpha, including Lpin2 and St3gal5, suggesting involvement of another transcription factor. Using chromatin immunoprecipitation, Lpin2 and St3gal5 were shown to be direct targets of PPARbeta/delta during fasting, whereas Aldh3a2 and Cpt2 were exclusive targets of PPARalpha. Binding of PPARbeta/delta to the Lpin2 and St3gal5 genes followed the plasma free fatty acid (FFA) concentration, consistent with activation of PPARbeta/delta by plasma FFAs. Subsequent experiments using transgenic and knockout mice for Angptl4, a potent stimulant of adipose tissue lipolysis, confirmed the stimulatory effect of plasma FFAs on Lpin2 and St3gal5 expression levels via PPARbeta/delta. In contrast, the data did not support activation of PPARalpha by plasma FFAs. The results identify Lpin2 and St3gal5 as novel PPARbeta/delta target genes and show that upregulation of gene expression by PPARbeta/delta is sensitive to plasma FFA levels. In contrast, this is not the case for PPARalpha, revealing a novel mechanism for functional differentiation between PPARs. PMID- 19805518 TI - S-adenosyl homocysteine hydrolase is required for Myc-induced mRNA cap methylation, protein synthesis, and cell proliferation. AB - The c-Myc proto-oncogene promotes mRNA cap methylation, which is essential for almost all mRNA translation. The mRNA cap methylation reaction produces an inhibitory byproduct, S-adenosyl homocysteine. Here we report that Myc promotes upregulation of S-adenosyl homocysteine hydrolase (SAHH), an enzyme which hydrolyzes S-adenosyl homocysteine, thus neutralizing its inhibitory effects, and this is required for c-Myc-induced mRNA cap methylation. c-Myc-induced mRNA cap methylation was repressed by inhibiting the expression or activity of SAHH, whereas the same treatments did not have a significant effect on c-Myc-induced transcription or other c-Myc-dependent methylation events. The selective inhibition of mRNA cap methylation afforded by SAHH repression revealed that c Myc-induced cap methylation could be correlated with the core c-Myc functions of protein synthesis, cell proliferation, and cell transformation. PMID- 19805519 TI - Bim upregulation by histone deacetylase inhibitors mediates interactions with the Bcl-2 antagonist ABT-737: evidence for distinct roles for Bcl-2, Bcl-xL, and Mcl 1. AB - The Bcl-2 antagonist ABT-737 kills transformed cells in association with displacement of Bim from Bcl-2. The histone deactetylase (HDAC) inhibitor suberoyl bis-hydroxamic acid (SBHA) was employed to determine whether and by what mechanism ABT-737 might interact with agents that upregulate Bim. Expression profiling of BH3-only proteins indicated that SBHA increased Bim, Puma, and Noxa expression, while SBHA concentrations that upregulated Bim significantly potentiated ABT-737 lethality. Concordance between SBHA-mediated Bim upregulation and interactions with ABT-737 was observed in various human leukemia and myeloma cells. SBHA-induced Bim was largely sequestered by Bcl-2 and Bcl-x(L), rather than Mcl-1; ABT-737 attenuated these interactions, thereby triggering Bak/Bax activation and mitochondrial outer membrane permeabilization. Knockdown of Bim (but not Puma or Noxa) by shRNA or ectopic overexpression of Bcl-2, Bcl-x(L), or Mcl-1 diminished Bax/Bak activation and apoptosis. Notably, ectopic expression of these antiapoptotic proteins disabled death signaling by sequestering different proapoptotic proteins, i.e., Bim by Bcl-2, both Bim and Bak by Bcl-x(L), and Bak by Mcl-1. Together, these findings indicate that HDAC inhibitor-inducible Bim is primarily neutralized by Bcl-2 and Bcl-x(L), thus providing a mechanistic framework by which Bcl-2 antagonists potentiate the lethality of agents, such as HDAC inhibitors, which upregulate Bim. PMID- 19805520 TI - Human SNF5/INI1, a component of the human SWI/SNF chromatin remodeling complex, promotes nucleotide excision repair by influencing ATM recruitment and downstream H2AX phosphorylation. AB - Recent studies have implicated the role of the SWI/SNF ATP-dependent chromatin remodeling complex in nuclear excision repair (NER), but the mechanism of its function has remained elusive. Here, we show that the human SWI/SNF component human SNF5 (hSNF5) interacts with UV damage recognition factor XPC and colocalizes with XPC at the damage site. Inactivation of hSNF5 did not affect the recruitment of XPC but affected the recruitment of ATM checkpoint kinase to the damage site and ATM activation by phosphorylation. Consequently, hSNF5 deficiency resulted in a defect in H2AX and BRCA1 phosphorylation at the damage site. However, recruitment of ATR checkpoint kinase to the damage site was not affected by hSNF5 deficiency, supporting that hSNF5 functions downstream of ATR. Additionally, ATM/ATR-mediated Chk2/Chk1 phosphorylation was not affected in hSNF5-depleted cells in response to UV irradiation, suggesting that the cell cycle checkpoint is intact in these cells. Taken together, the results indicate that the SWI/SNF complex associates with XPC at the damage site and thereby facilitates the access of ATM, which in turn promotes H2AX and BRCA1 phosphorylation. We propose that the SWI/SNF chromatin remodeling function is utilized to increase the DNA accessibility of NER machinery and checkpoint factors at the damage site, which influences NER and ensures genomic integrity. PMID- 19805521 TI - Hepatocyte nuclear factor 4alpha, a key factor for homeostasis, cell architecture, and barrier function of the adult intestinal epithelium. AB - Hepatocyte nuclear factor 4alpha (HNF-4alpha) is a transcription factor which is highly expressed in the intestinal epithelium from duodenum to colon and from crypt to villus. The homeostasis of this constantly renewing epithelium relies on an integrated control of proliferation, differentiation, and apoptosis, as well as on the functional architecture of the epithelial cells. In order to determine the consequences of HNF-4alpha loss in the adult intestinal epithelium, we used a tamoxifen-inducible Cre-loxP system to inactivate the Hnf-4a gene. In the intestines of adult mice, loss of HNF-4alpha led to an increased proliferation in crypts and to an increased expression of several genes controlled by the Wnt/beta catenin system. This control of the Wnt/beta-catenin signaling pathway by HNF 4alpha was confirmed in vitro. Cell lineage was affected, as indicated by an increased number of goblet cells and an impairment of enterocyte and enteroendocrine cell maturation. In the absence of HNF-4alpha, cell-cell junctions were destabilized and paracellular intestinal permeability increased. Our results showed that HNF-4alpha modulates Wnt/beta-catenin signaling and controls intestinal epithelium homeostasis, cell function, and cell architecture. This study indicates that HNF-4alpha regulates the intestinal balance between proliferation and differentiation, and we hypothesize that it might act as a tumor suppressor. PMID- 19805522 TI - Semaphorin 4D signaling requires the recruitment of phospholipase C gamma into the plexin-B1 receptor complex. AB - The semaphorin 4D (Sema4D) receptor plexin-B1 constitutively interacts with particular Rho guanine nucleotide exchange factors (RhoGEFs) and thereby mediates Sema4D-induced RhoA activation, a process which involves the tyrosine phosphorylation of plexin-B1 by ErbB-2. It is, however, unknown how plexin-B1 phosphorylation regulates RhoGEF activity. We show here that activation of plexin B1 by Sema4D and its subsequent tyrosine phosphorylation creates docking sites for the SH2 domains of phospholipase Cgamma (PLCgamma). PLCgamma is thereby recruited into the plexin-B1 receptor complex and via its SH3 domain activates the Rho guanine nucleotide exchange factor PDZ-RhoGEF. PLCgamma-dependent RhoGEF activation is independent of its lipase activity. The recruitment of PLCgamma has no effect on the R-Ras GTPase-activating protein activity of plexin-B1 but is required for Sema4D-induced axonal growth cone collapse as well as for the promigratory effects of Sema4D on cancer cells. These data demonstrate a novel nonenzymatic function of PLCgamma as an important mechanism of plexin-mediated signaling which links tyrosine phosphorylation of plexin-B1 to the regulation of a RhoGEF protein and downstream cellular processes. PMID- 19805524 TI - Molecular characterization of EmABP, an apolipoprotein A-I binding protein secreted by the Echinococcus multilocularis metacestode. AB - Cestodes are unable to synthesize de novo most of their own membrane lipids, including cholesterol, and have to take them up from the host during an infection. The underlying molecular mechanisms are so far unknown. Here we report the identification and characterization of a novel gene, Emabp, which is expressed by larval stages and adults of the fox tapeworm Echinococcus multilocularis. The encoded protein, EmABP, displays significant homologies to apolipoprotein A-I binding protein (AI-BP) of mammalian origin and to metazoan YjeF_N domain proteins. Like mammalian AI-BP, EmABP carries an export-directing signal sequence which is absent in predicted AI-BP orthologs from the related flatworms Schistosoma japonicum and Schmidtea mediterranea. Using a specific antibody and immunoprecipitation techniques, we demonstrate that EmABP is secreted into the extraparasitic environment and into the hydatid fluid of in vitro-cultivated metacestode vesicles. Furthermore, we show that apolipoprotein A I (apoA-I), a major constituent of cholesterol-transporting high-density lipoproteins, is present in hydatid fluid. By pulldown experiments, we demonstrate that recombinantly expressed, purified EmABP interacts with purified human apoA-I and is able to precipitate apoA-I from human serum. On the basis of these features and the suggested function of AI-BP in cholesterol transport in higher eukaryotes, we propose a role for EmABP in cholesterol and lipid uptake mechanisms of larval E. multilocularis. PMID- 19805523 TI - Instantaneous within-patient diversity of Pseudomonas aeruginosa quorum-sensing populations from cystic fibrosis lung infections. AB - In the opportunistic pathogen Pseudomonas aeruginosa, acyl-homoserine lactone (acyl-HSL) quorum sensing (QS) regulates biofilm formation and expression of many extracellular virulence factors. Curiously, QS-deficient variants, often carrying mutations in the central QS regulator LasR, are frequently isolated from infections, particularly from cystic fibrosis (CF) lung infections. Very little is known about the proportion and diversity of these QS variants in individual infections. Such information is desirable to better understand the selective forces that drive the evolution of QS phenotypes, including social cheating and innate (nonsocial) benefits. To obtain insight into the instantaneous within patient diversity of QS, we assayed a panel of 135 concurrent P. aeruginosa isolates from eight different adult CF patients (9 to 20 isolates per patient) for various QS-controlled phenotypes. Most patients contained complex mixtures of QS-proficient and -deficient isolates. Among all patients, deficiency in individual phenotypes ranged from 0 to about 90%. Acyl-HSL, sequencing, and complementation analyses of variants with global loss-of-function phenotypes revealed dependency upon the central QS circuitry genes lasR, lasI, and rhlI. Deficient and proficient isolates were clonally related, implying evolution from a common ancestor in vivo. Our results show that the diversity of QS types is high within and among patients, suggesting diverse selection pressures in the CF lung. A single selective mechanism, be it of a social or nonsocial nature, is unlikely to account for such heterogeneity. The observed diversity also shows that conclusions about the properties of P. aeruginosa QS populations in individual CF infections cannot be drawn from the characterization of one or a few selected isolates. PMID- 19805525 TI - A heterologous helper T-cell epitope enhances the immunogenicity of a multiple antigenic-peptide vaccine targeting the cryptic loop-neutralizing determinant of Bacillus anthracis protective antigen. AB - We previously showed that a multiple antigenic peptide (MAP) displaying amino acids (aa) 305 to 319 from the 2beta2-2beta3 loop of protective antigen (PA) can elicit high-titered antibody that neutralizes lethal toxin (LeTx) in vitro and that this loop-neutralizing determinant (LND) specificity is absent in PA-immune rabbits. Some immune rabbits were, however, nonresponders to the MAP. We hypothesized that the immunogen elicited suboptimal major histocompatibility complex (MHC) class II-restricted T-cell help and that introduction of a functional helper T-cell epitope would increase MHC-restricted responsiveness and the magnitude and affinity of the antibody responses. In the current study, we characterized the T- and B-cell responses to LND peptides in mice, then designed second-generation MAP immunogens for eliciting LND-specific immunity, and tested them in rabbits. The 305-319 sequence was devoid of helper T-cell epitopes in three strains of mice; however, a T-B peptide comprising aa 305 to 319, colinearly synthesized with the P30 helper epitope of tetanus toxin, elicited robust LeTx-neutralizing immunity in mice. T-B MAPs displaying B-cell epitopes 304 to 319 (MAP304) or 305 to 319 (MAP305) elicited high-titer, durable antibody responses in rabbits which exhibited potent neutralization of LeTx in vitro. All MAP304-immune rabbits demonstrated neutralization titers exceeding that of hyperimmune sera of rabbits immunized with PA in Freund's adjuvant, with peak neutralization titers 23-, 6-, and 3-fold higher than that of the PA antiserum. Overall, immunization with MAPs containing the P30 epitope elicited higher antibody and toxin neutralization titers and peptide-specific affinity than immunization with an LND MAP lacking a helper epitope. P30-containing MAP304 represents a promising LND-specific vaccine for anthrax. PMID- 19805526 TI - Inhibition of erythrocyte invasion and Plasmodium falciparum merozoite surface protein 1 processing by human immunoglobulin G1 (IgG1) and IgG3 antibodies. AB - Antigen-specific antibodies (Abs) to the 19-kDa carboxy-terminal region of Plasmodium falciparum merozoite surface protein 1 (MSP1(19)) play an important role in protective immunity to malaria. Mouse monoclonal Abs (MAbs) 12.10 and 12.8 recognizing MSP1(19) can inhibit red cell invasion by interfering with MSP1 processing on the merozoite surface. We show here that this ability is dependent on the intact Ab since Fab and F(ab')(2) fragments derived from MAb 12.10, although capable of binding MSP1 with high affinity and competing with the intact antibody for binding to MSP1, were unable to inhibit erythrocyte invasion or MSP1 processing. The DNA sequences of the variable (V) regions of both MAbs 12.8 and 12.10 were obtained, and partial amino acid sequences of the same regions were confirmed by mass spectrometry. Human chimeric Abs constructed by using these sequences, which combine the original mouse V regions with human gamma1 and gamma3 constant regions, retain the ability to bind to both parasites and recombinant MSP1(19), and both chimeric human immunoglobulin G1s (IgG1s) were at least as good at inhibiting erythrocyte invasion as the parental murine MAbs 12.8 and 12.10. Furthermore, the human chimeric Abs of the IgG1 class (but not the corresponding human IgG3), induced significant NADPH-mediated oxidative bursts and degranulation from human neutrophils. These chimeric human Abs will enable investigators to examine the role of human Fcgamma receptors in immunity to malaria using a transgenic parasite and mouse model and may prove useful in humans for neutralizing parasites as an adjunct to antimalarial drug therapy. PMID- 19805527 TI - Inescapable need for neutrophils as mediators of cellular innate immunity to acute Pseudomonas aeruginosa pneumonia. AB - Pseudomonas aeruginosa is a leading cause of pneumonia, and many components of the innate immune system have been proposed to exert important effects in preventing lung infection. However, a vigorous experimental system to identify an overriding, key effector mediating innate immunity to lung infection has not been utilized. As many of the important components of innate immunity are involved in recruitment and activation of polymorphonuclear neutrophils (PMNs) to infected tissues, we hypothesized that the cells and factors needed for their proper recruitment to the lung comprised the major mediators of innate immunity. In neutropenic mice, intranasal (i.n.) doses of P. aeruginosa as low as 10 to 100 CFU/mouse produced a fatal lung infection, compared with 10(7) to >10(8) CFU for nonneutropenic mice. There was only a very modest increased mortality in mice lacking mature lymphocytes and no increased mortality in mice depleted of alveolar macrophages when administered i.n. P. aeruginosa. Recombinant mouse granulocyte colony-stimulating factor increased survival of neutropenic mice after i.n. P. aeruginosa inoculation. MyD88(-/-) mice, which cannot recruit PMNs to the lungs, were highly susceptible to fatal P. aeruginosa lung infection, with bacterial doses of <120 CFU being lethal. Activation of a MyD88-independent pathway for PMN recruitment to the lungs in MyD88(-/-) mice resulted in enhanced protection against P. aeruginosa lung infection. Overall, in the absence of PMNs, mice cannot resist P. aeruginosa lung infection from extremely small bacterial doses. There is an inescapable requirement for local PMN recruitment and activation to mediate innate immunity to P. aeruginosa lung infection. PMID- 19805528 TI - The glycosylphosphatidylinositol-anchored protease Sap9 modulates the interaction of Candida albicans with human neutrophils. AB - Human polymorphonuclear neutrophils (PMNs) play a major role in the immune defense against invasive Candida albicans infection. This fungal pathogen produces a set of aspartic proteases that directly contributes to virulence properties such as adhesion, tissue invasion, and immune evasion. We show here that, in contrast to other secreted proteases, the cell surface-associated isoform Sap9 has a major impact on the recognition of C. albicans by PMNs. SAP9 is required for the induction of PMN chemotaxis toward C. albicans filaments, an essential prerequisite of effective PMN activation. Furthermore, deletion of SAP9 leads to a mitigated release of reactive oxygen intermediates (ROI) in human PMNs and decreases C. albicans-induced apoptosis triggered by ROI formation. In confrontation assays, killing of a SAP9 deletion mutant is reduced in comparison to wild-type C. albicans. These data clearly implicate Sap9 protease activity in the initiation of protective innate immunity and suggest novel molecular mechanisms in C. albicans-host interaction leading to neutrophil activation. PMID- 19805529 TI - The secreted esterase of group a streptococcus is important for invasive skin infection and dissemination in mice. AB - Virulence factors regulated by the CovRS/CsrRS two-component gene regulatory system contribute to the invasive diseases caused by group A Streptococcus (GAS). To determine whether the streptococcal secreted esterase (Sse), an antigen that protects against subcutaneous GAS infection, is one of these virulence factors, we investigated the phenotype of a nonpolar sse deletion mutant strain (Deltasse). In addition, we examined the effects of covS mutation on sse expression. As assessed using a mouse model of subcutaneous infection, the virulence of the Deltasse strain is attenuated and the overall pathology is reduced. Furthermore, GAS was detected in the blood and spleens from mice subcutaneously infected with the parental strain, whereas mice subcutaneously infected with the Deltasse strain had no GAS present in their blood and spleens. The ability of the mutant to survive in the subcutis of mice appeared to be compromised. The growth of the Deltasse strain in rich and chemically defined media and nonimmune human blood and sera was slower than that of the wild-type strain. Complementation restored the phenotype of the Deltasse strain to that of the wild-type strain. The wild-type, Deltasse, and complement strains had no detectable SpeB activity. Expression of Sse is negatively controlled by CovRS. These findings suggest that Sse is a CovRS-regulated virulence factor that is important for the virulence of GAS in subcutaneous infection and plays an important role in severe soft tissue infections and systemic dissemination of GAS from the skin. PMID- 19805530 TI - Toll-like receptors 2 and 4 contribute to sepsis-induced depletion of spleen dendritic cells. AB - Depletion of dendritic cells (DC) in secondary lymphoid organs is a hallmark of sepsis-induced immune dysfunction. In this setting, we investigated if Toll-like receptor (TLR)-dependent signaling might modulate the maturation process and the survival of DC. Using a model of sublethal polymicrobial sepsis induced by cecal ligation and puncture, we investigated the quantitative and functional features of spleen DC in wild-type, TLR2(-/-), TLR4(-/-), and TLR2(-/-) TLR4(-/-) mice. By 24 h, a decrease in the relative percentage of CD11c(high) spleen DC occurred in wild-type mice but was prevented in TLR2(-/-), TLR4(-/-), and TLR2(-/-) TLR4(-/-) mice. In wild-type mice, sepsis dramatically affected both CD11c(+) CD8alpha(+) and CD11c(+) CD8alpha(-) subsets. In all three types of knockout mice studied, the CD11c(+) CD8alpha(+) subset followed a depletion pattern similar to that for wild-type mice. In contrast, the loss of CD11c(+) CD8alpha(-) cells was attenuated in TLR2(-/-) and TLR4(-/-) mice and completely prevented in TLR2(-/-) TLR4(-/-) mice. Accordingly, apoptosis of spleen DC was increased in septic wild type mice and inhibited in knockout mice. In addition we characterized the functional features of spleen DC obtained from septic mice. As shown by increased expression of major histocompatibility complex class II and CD86, polymicrobial sepsis induced maturation of DC, with subsequent increased capacity to prime T lymphocytes, similarly in wild-type and knockout mice. In response to CpG DNA stimulation, production of interleukin-12 was equally impaired in DC obtained from wild-type and knockout septic mice. In conclusion, although dispensable for the DC maturation process, TLR2 and TLR4 are involved in the mechanisms leading to depletion of spleen DC following polymicrobial sepsis. PMID- 19805531 TI - The Sca2 autotransporter protein from Rickettsia conorii is sufficient to mediate adherence to and invasion of cultured mammalian cells. AB - Obligate intracellular bacteria of the genus Rickettsia must adhere to and invade the host endothelium in order to establish an infection. These processes require the interaction of rickettsial surface proteins with mammalian host cell receptors. A previous bioinformatic analysis of sequenced rickettsial species identified a family of at least 17 predicted "surface cell antigen" (sca) genes whose products resemble autotransporter proteins. Two members of this family, rOmpA and rOmpB of spotted fever group (SFG) rickettsiae have been identified as adhesion and invasion factors, respectively; however, little is known about the putative functions of the other sca gene products. An intact sca2 gene is found in the majority of pathogenic SFG rickettsiae and, due to its sequence conservation among these species, we predict that Sca2 may play an important function at the rickettsial surface. Here we have shown that sca2 is transcribed and expressed in Rickettsia conorii and have used a heterologous gain-of-function assay in E. coli to determine the putative role of Sca2. Using this system, we have demonstrated that expression of Sca2 at the outer membrane of nonadherent, noninvasive E. coli is sufficient to mediate adherence to and invasion of a panel of mammalian cells, including endothelial cells. Furthermore, soluble Sca2 protein is capable of diminishing R. conorii invasion of cultured mammalian cells. This is the first evidence that Sca2 participates in the interaction between SFG rickettsiae and host cells and suggests that in addition to other surface proteins, Sca2 may play a critical role in rickettsial pathogenesis. PMID- 19805532 TI - Persistent infection contributes to heterologous protective immunity against fatal ehrlichiosis. AB - Human monocytotropic ehrlichiosis (HME), an emerging and often life-threatening tick-transmitted disease, is caused by the obligately intracellular bacterium Ehrlichia chaffeensis. HME is modeled in C57BL/6 mice using Ehrlichia muris, which causes persistent infection, and Ixodes ovatus Ehrlichia (IOE), which is either acutely lethal or sublethal depending on the dose and route of inoculation. A persistent primary E. muris infection, but not a sublethal IOE infection, protects mice against an ordinarily lethal secondary IOE challenge. In the present study, we determined the role of persistent infection in maintenance of protective memory immune responses. E. muris-infected mice were treated with doxycycline or left untreated and then challenged with an ordinarily lethal dose of IOE. Compared to E. muris-primed mice treated with doxycycline, untreated mice persistently infected with E. muris had significantly greater numbers of antigen specific gamma interferon-producing splenic memory T cells, significant expansion of CD4(+) CD25(+) T regulatory cells, and production of transforming growth factor beta1 in the spleen. Importantly, E. muris-primed mice treated with doxycycline showed significantly greater susceptibility to challenge infection with IOE compared to untreated mice persistently infected with E. muris. The study indicated that persistent ehrlichial infection contributes to heterologous protection by stimulating the maintenance of memory T-cell responses. PMID- 19805533 TI - C3H male mice with severe combined immunodeficiency cannot clear a urethral infection with a human serovar of Chlamydia trachomatis. AB - The pathogenesis of an infection of the male genitourinary tract of mice with a human serovar of Chlamydia trachomatis has not been characterized. To establish a new model, we inoculated C3H/HeN (H-2(k)) mice in the meatus urethra with C. trachomatis serovar D. To determine the 50% infectious dose (ID(50)), male mice were inoculated with doses ranging from 10(2) to 10(6) inclusion-forming units (IFU). The mice were euthanized 10 days post infection (p.i.), and the urethra, bladder, epididimydes, and testes were cultured for Chlamydia. Positive cultures were obtained from the urethra, urinary bladder, and epididimydes, and the ID(50) was determined to be 5 x 10(4) IFU/mouse. Subsequently, to characterize the course of the infection, wild-type (WT) and C3H animals with severe combined immunodeficiency (SCID animals) were inoculated with 10(6) IFU/mouse (20 times the ID(50)). In the WT mice, the infection peaked in the second week, and by 42 days p.i., it was cleared. In contrast, most of the SCID mice continued to have positive cultures at 60 days p.i. C. trachomatis-specific antibodies were first detected in WT animals' sera at 21 days p.i. and increased until 42 days p.i. The immunoglobulin G2a (IgG2a) titers were 32-fold higher than those of IgG1, indicative of a Th1-biased immune response. A lymphoproliferative assay using splenocytes showed a significant cell-mediated immune response in the WT mice. As expected, no humoral or cell-mediated immune responses were observed in the SCID animals. In conclusion, inoculation of WT male mice in the meatus urethra with a human serovar of C. trachomatis resulted in a limited infection mainly localized to the lower genitourinary tract. On the other hand, SCID animals could not clear the infection, suggesting that in male mice, the adaptive immune response is necessary to control an infection with a C. trachomatis human serovar. PMID- 19805534 TI - Spa15 of Shigella flexneri is secreted through the type III secretion system and prevents staurosporine-induced apoptosis. AB - Shigella flexneri is a gram-negative, facultative intracellular pathogen that invades the colonic epithelium and causes bacillary dysentery. We previously demonstrated that S. flexneri inhibits staurosporine-induced apoptosis in infected epithelial cells and that a DeltamxiE mutant is unable to inhibit apoptosis. Therefore, we hypothesized that an MxiE-regulated gene was responsible for protection of epithelial cells from apoptosis. Analysis of all MxiE-regulated genes yielded no mutants that lacked the ability to prevent apoptosis. Spa15, which is defined as a type III secretion system chaperone, was analyzed since it associates with MxiE. A Deltaspa15 mutant was unable to prevent staurosporine induced apoptosis. C-terminal hemagglutinin-tagged spa15 was secreted by S. flexneri within 2 h in the Congo red secretion assay, and secretion was dependent on the type III secretion system. Spa15 was also secreted by Shigella in infected epithelial cells, as verified by immunofluorescence analysis. Spa15 secretion was decreased in the DeltamxiE mutant, which demonstrates why this mutant is unable to prevent staurosporine-induced apoptosis. Our data are the first to show that Spa15 is secreted in a type III secretion system-dependent fashion, and the absence of Spa15 in the Deltaspa15 mutant results in the loss of protection from staurosporine-induced apoptosis in epithelial cells. Thus, Spa15 contributes to the intracellular survival of Shigella by blocking apoptosis in the infected host cell. PMID- 19805535 TI - Critical role for interleukin-1beta (IL-1beta) during Chlamydia muridarum genital infection and bacterial replication-independent secretion of IL-1beta in mouse macrophages. AB - Recent findings have implicated interleukin-1beta (IL-1beta) as an important mediator of the inflammatory response in the female genital tract during chlamydial infection. But how IL-1beta is produced and its specific role in infection and pathology are unclear. Therefore, our goal was to determine the functional consequences and cellular sources of IL-1beta expression during a chlamydial genital infection. In the present study, IL-1beta(-/-) mice exhibited delayed chlamydial clearance and decreased frequency of hydrosalpinx compared to wild-type (WT) mice, implying an important role for IL-1beta both in the clearance of infection and in the mediation of oviduct pathology. At the peak of IL-1beta secretion in WT mice, the major producers of IL-1beta in vivo are F4/80(+) macrophages and GR-1(+) neutrophils, but not CD45(-) epithelial cells. Although elicited mouse macrophages infected with Chlamydia muridarum in vitro secrete minimal IL-1beta, in vitro prestimulation of macrophages by Toll-like receptor (TLR) ligands such as lipopolysaccharide (LPS) purified from Escherichia coli or C. trachomatis L2 prior to infection greatly enhanced secretion of IL 1beta from these cells. By using LPS-primed macrophages as a model system, it was determined that IL-1beta secretion was dependent on caspase-1, potassium efflux, and the activity of serine proteases. Significantly, chlamydia-induced IL-1beta secretion in macrophages required bacterial viability but not growth. Our findings demonstrate that IL-1beta secreted by macrophages and neutrophils has important effects in vivo during chlamydial infection. Additionally, prestimulation of macrophages by chlamydial TLR ligands may account for the elevated levels of pro-IL-1beta mRNA observed in vivo in this cell type. PMID- 19805536 TI - The long-lived nature of clostridium perfringens iota toxin in mammalian cells induces delayed apoptosis. AB - Mono-ADP ribosylation of actin by bacterial toxins, such as Clostridium perfringens iota or Clostridium botulinum C2 toxins, results in rapid depolymerization of actin filaments and cell rounding. Here we report that treatment of African green monkey kidney (Vero) cells with iota toxin resulted in delayed caspase-dependent death. Unmodified actin did not reappear in toxin treated cells, and enzyme-active toxin was detectable in the cytosol for at least 24 h. C2 toxin showed comparable, long-lived effects in cells, while a C2 toxin control lacking ADP-ribosyltransferase activity did not induce cell death. To address whether the remarkable stability of the iota and C2 toxins in cytosol was crucial for inducing cell death, we treated cells with C/SpvB, the catalytic domain of Salmonella enterica SpvB. Although C/SpvB also mono-ADP ribosylates actin as do the iota and C2 toxins, cells treated with a cell-permeating C/SpvB fusion toxin became rounded but recovered and remained viable. Moreover, unmodified actin reappeared in these cells, and ADP-ribosyltransferase activity due to C/SpvB was not detectable in the cytosol after 24 h, a result most likely due to degradation of C/SpvB. Repeated application of C/SpvB prevented recovery of cells and reappearance of unmodified actin. In conclusion, a complete but transient ADP ribosylation of actin was not sufficient to trigger apoptosis, implying that long-term stability of actin-ADP-ribosylating toxins, such as iota and C2, in the cytosol is crucial for inducing delayed, caspase-dependent cell death. PMID- 19805537 TI - Development and application of new mouse models to study the pathogenesis of Clostridium perfringens type C Enterotoxemias. AB - Clostridium perfringens type C isolates cause enterotoxemias and enteritis in humans and livestock. While the major disease signs and lesions of type C disease are usually attributed to beta toxin (CPB), these bacteria typically produce several different lethal toxins. Since understanding of disease pathogenesis and development of improved vaccines is hindered by the lack of small animal models mimicking the lethality caused by type C isolates, in this study we developed two mouse models of C. perfringens type C-induced lethality. When inoculated into BALB/c mice by intragastric gavage, 7 of 14 type C isolates were lethal, whereas when inoculated intraduodenally, these strains were all lethal in these mice. Clinical signs in intragastrically and intraduodenally challenged mice were similar and included respiratory distress, abdominal distension, and neurological alterations. At necropsy, the small, and occasionally the large, intestine was dilated and gas filled in most mice developing a clinical response. Histological changes in the gut were relatively mild, consisting of attenuation of the mucosa with villus blunting. Inactivation of the CPB-encoding gene rendered the highly virulent type C strain CN3685 avirulent in the intragastric model and nearly nonlethal in the intraduodenal model. In contrast, inactivation of the genes encoding alpha toxin and perfringolysin O only slightly decreased the lethality of CN3685. Mice could be protected against lethality by intravenous passive immunization with a CPB antibody prior to intragastric challenge. This study proves that CPB is a major contributor to the systemic effects of type C infections and provides new mouse models for investigating the pathogenesis of type C-induced lethality. PMID- 19805538 TI - Regulated delayed expression of rfaH in an attenuated Salmonella enterica serovar typhimurium vaccine enhances immunogenicity of outer membrane proteins and a heterologous antigen. AB - RfaH is a transcriptional antiterminator that reduces the polarity of long operons encoding secreted and surface-associated cell components of Salmonella enterica serovar Typhimurium, including O antigen and lipopolysaccharide core sugars. A DeltarfaH mutant strain is attenuated in mice (50% lethal dose [LD(50)], >10(8) CFU). To examine the potential for using rfaH in conjunction with other attenuating mutations, we designed a series of strains in which we replaced the native rfaH promoter with the tightly regulated arabinose-dependent araC P(BAD) promoter so that rfaH expression was dependent on exogenously supplied arabinose provided during in vitro growth. Following colonization of host lymphoid tissues, where arabinose was not available, the P(BAD) promoter was no longer active and rfaH was not expressed. In the absence of RfaH, O antigen and core sugars were not synthesized. We constructed three mutant strains that expressed different levels of RfaH by altering the ribosome-binding sequence and start codon. One mutation, DeltaP(rfaH178), was introduced into the attenuated vaccine strain chi9241 (DeltapabA DeltapabB DeltaasdA) expressing the pneumococcal surface protein PspA from an Asd(+) balanced-lethal plasmid. Mice immunized with this strain and boosted 4 weeks later induced higher levels of serum immunoglobulin G specific for PspA and for outer membrane proteins from other enteric bacteria than either an isogenic DeltarfaH derivative or the isogenic RfaH(+) parent. Eight weeks after primary oral immunization, mice were challenged with 200 LD(50) of virulent Streptococcus pneumoniae WU2. Immunization with DeltaP(rfaH178) mutant strains led to increased levels of protection compared to that of the parent chi9241 and of a DeltarfaH derivative of chi9241. PMID- 19805539 TI - Leptospira interrogans binds to human cell surface receptors including proteoglycans. AB - Leptospirosis is a global public health problem, primarily in the tropical developing world. The pathogenic mechanisms of the causative agents, several members of the genus Leptospira, have been underinvestigated. The exception to this trend has been the demonstration of the binding of pathogenic leptospires to the extracellular matrix (ECM) and its components. In this work, interactions of Leptospira interrogans bacteria with mammalian cells, rather than the ECM, were examined. The bacteria bound more efficiently to the cells than to the ECM, and a portion of this cell-binding activity was attributable to attachment to glycosaminoglycan (GAG) chains of proteoglycans (PGs). Chondroitin sulfate B PGs appeared to be the primary targets of L. interrogans attachment, while heparan sulfate PGs were much less important. Inhibition of GAG/PG-mediated attachment resulted in partial inhibition of bacterial attachment, suggesting that additional receptors for L. interrogans await identification. GAG binding may participate in the pathogenesis of leptospirosis within the host animal. In addition, because GAGs are expressed on the luminal aspects of epithelial cells in the proximal tubules of the kidneys, this activity may play a role in targeting the bacteria to this critical site. Because GAGs are shed in the urine, GAG binding may also be important for transmission to new hosts through the environment. PMID- 19805540 TI - Saliva enables the antimicrobial activity of LL-37 in the presence of proteases of Porphyromonas gingivalis. AB - Proteolysis is a common microbial virulence mechanism that enables the destruction of host tissue and evasion from host defense mechanisms. Antimicrobial peptides, also known as host defense peptides, are effector molecules of the innate immunity that demonstrate a broad range of antimicrobial and immunoregulatory activities. Deficiency of the human LL-37 antimicrobial peptide was previously correlated with severe periodontal disease. Porphyromonas gingivalis, the major pathogen associated with periodontitis, is highly proteolytic. In this study, P. gingivalis was found capable of degrading LL-37 by utilizing its arginine-specific gingipains. Saliva collected from volunteers with a healthy periodontium protected LL-37 from proteolysis by P. gingivalis. Salivary protection of LL-37 was heat resistant and specific and enabled LL-37 to inhibit growth of Escherichia coli in the presence of the P. gingivalis proteases. Previously, saliva and other body fluids have been shown to inhibit the antimicrobial activity of LL-37. Here we demonstrate that at a cost of a small reduction in the bactericidal activity of LL-37, saliva enables the antibacterial activity of LL-37 despite the presence of proteases secreted by the main periodontopathogen. PMID- 19805541 TI - Enterococcus faecalis capsular polysaccharide serotypes C and D and their contributions to host innate immune evasion. AB - It has become increasingly difficult to treat infections caused by Enterococcus faecalis due to its high levels of intrinsic and acquired antibiotic resistance. However, few studies have explored the mechanisms that E. faecalis employs to circumvent the host innate immune response and establish infection. Capsular polysaccharides are important virulence factors that are associated with innate immune evasion. We demonstrate, using cultured macrophages (RAW 264.7), that capsule-producing E. faecalis strains of either serotype C or D are more resistant to complement-mediated opsonophagocytosis than unencapsulated strains. We show that differences in opsonophagocytosis are not due to variations in C3 deposition but are due to the ability of capsule to mask bound C3 from detection on the surface of E. faecalis. Similarly, E. faecalis capsule masks lipoteichoic acid from detection, which correlates with decreased tumor necrosis factor alpha production by cultured macrophages in the presence of encapsulated strains compared to that in the presence of unencapsulated strains. Our studies confirm the important role of the capsule as a virulence factor of E. faecalis and provide several mechanisms by which the presence of the capsule influences evasion of the innate immune response and suggest that the capsule could be a potential target for developing alternative therapies to treat E. faecalis infections. PMID- 19805542 TI - CD23 mediates antimycobacterial activity of human macrophages. AB - Engagement of surface receptors contributes to the antimicrobial activity of human immune cells. We show here that infection of human monocyte-derived macrophages (MDM) with live Mycobacterium avium induced the expression of CD23 on their membrane. Subsequent cross-linking of surface CD23 by appropriate ligands induced a dose-dependent antibacterial activity of MDM and the elimination of most infected cells. The stimulation of inducible nitric oxide synthase-dependent generation of NO from MDM after CD23 activation played a major role during their anti-M. avium activity. CD23 activation also induced tumor necrosis factor alpha (TNF-alpha) production from MDM. Mycobacteria reduction was partially inhibited by the addition of neutralizing anti-TNF-alpha antibody to cell cultures without affecting NO levels, which suggested the role of this cytokine for optimal antimicrobial activity. Finally, interleukin-10, a Th2 cytokine known to downregulate CD23 pathway, is shown to decrease NO generation and mycobacteria elimination by macrophages. Therefore, (i) infection with M. avium promotes functional surface CD23 expression on human macrophages and (ii) subsequent signaling of this molecule contributes to the antimicrobial activity of these cells through an NO- and TNF-alpha-dependent pathway. This study reveals a new human immune response mechanism to counter mycobacterial infection involving CD23 and its related ligands. PMID- 19805543 TI - Organization of cytochrome P450 enzymes involved in sex steroid synthesis: PROTEIN-PROTEIN INTERACTIONS IN LIPID MEMBRANES. AB - Mounting evidence underscores the importance of protein-protein interactions in the functional regulation of drug-metabolizing P450s, but few studies have been conducted in membrane environments, and none have examined P450s catalyzing sex steroid synthesis. Here we report specific protein-protein interactions for full length, human, wild type steroidogenic cytochrome P450 (P450, CYP) enzymes: 17alpha-hydroxylase/17,20-lyase (P450c17, CYP17) and aromatase (P450arom, CYP19), as well as their electron donor NADPH-cytochrome P450 oxidoreductase (CPR). Fluorescence resonance energy transfer (FRET)(3) in live cells, coupled with quartz crystal microbalance (QCM), and atomic force microscopy (AFM) studies on phosphatidyl choline +/- cholesterol (mammalian) biomimetic membranes were used to investigate steroidogenic P450 interactions. The FRET results in living cells demonstrated that both P450c17 and P450arom homodimerize but do not heterodimerize, although they each heterodimerize with CPR. The lack of heteroassociation between P450c17 and P450arom was confirmed by QCM, wherein neither enzyme bound a membrane saturated with the other. In contrast, the CPR bound readily to either P450c17- or P450arom-saturated surfaces. Interestingly, N terminally modified P450arom was stably incorporated and gave similar results to the wild type, although saturation was achieved with much less protein, suggesting that the putative transmembrane domain is not required for membrane association but for orientation. In fact, all of the proteins were remarkably stable in the membrane, such that high resolution AFM images were obtained, further supporting the formation of P450c17, P450arom, and CPR homodimers and oligomers in lipid bilayers. This unique combination of in vivo and in vitro studies has provided strong evidence for homodimerization and perhaps some higher order interactions for both P450c17 and P450arom. PMID- 19805544 TI - Bax activates endophilin B1 oligomerization and lipid membrane vesiculation. AB - Endophilins participate in membrane scission events that occur during endocytosis and intracellular organelle biogenesis through the combined activity of an N terminal BAR domain that interacts with membranes and a C-terminal SH3 domain that mediates protein binding. Endophilin B1 (Endo B1) was identified to bind Bax, a Bcl-2 family member that promotes apoptosis, through yeast two-hybrid protein screens. Although Endo B1 does not bind Bax in healthy cells, during apoptosis, Endo B1 interacts transiently with Bax and promotes cytochrome c release from mitochondria. To explore the molecular mechanism of action of Endo B1, we have analyzed its interaction with Bax in cell-free systems. Purified recombinant Endo B1 in solution displays a Stokes radius indicating a tetrameric quarternary structure. However, when incubated with purified Bax, it assembles into oligomers more than 4-fold greater in molecular weight. Although Endo B1 oligomerization is induced by Bax, Bax does not stably associate with the high molecular weight Endo B1 complex. Endo B1 oligomerization requires its C-terminal Src homology 3 domain and is not induced by Bcl-xL. Endo B1 combined with Bax reduces the size and changes the morphology of giant unilamellar vesicles by inducing massive vesiculation of liposomes. This activity of purified Bax protein to induce cell-free assembly of Endo B1 may reflect its activity in cells that regulates apoptosis and/or mitochondrial fusion. PMID- 19805545 TI - Noncatalytic function of ERK1/2 can promote Raf/MEK/ERK-mediated growth arrest signaling. AB - Kinase activity is known as the key biochemical property of MAPKs. Here, we report that ERK1/2 also utilizes its noncatalytic function to mediate certain signal transductions. Sustained activation of the Raf/MEK/ERK pathway induces growth arrest, accompanied by changes in cell cycle regulators (decreased retinoblastoma phosphorylation, E2F1 down-regulation, and/or p21(CIP1) up regulation) and cell type-specific changes in morphology and expression of c-Myc or RET in the human tumor lines LNCaP, U251, and TT. Ablation of ERK1/2 by RNA interference abrogated all these effects. However, active site-disabled ERK mutants (ERK1-K71R, ERK2-K52R, and ERK2-D147A), which competitively inhibit activation of endogenous ERK1/2, could not block Raf/MEK-induced growth arrest as well as changes in the cell cycle regulators, although they effectively blocked phosphorylation of the ERK1/2 catalytic activity readouts, p90(RSK) and ELK1, as well as the cell type-specific changes. Because this indicated a potential noncatalytic ERK1/2 function, we generated stable lines of the tumor cells in which both ERK1 and ERK2 were significantly knocked down, and we further investigated the possibility using rat-derived kinase-deficient ERK mutants (ERK2 K52R and ERK2-T183A/Y185F) that were not targeted by human small hairpin RNA. Indeed, ERK2-K52R selectively restored Raf-induced growth inhibitory signaling in ERK1/2-depleted cells, as manifested by regained cellular ability to undergo growth arrest and to control the cell cycle regulators without affecting c-Myc and morphology. However, ERK2-T183A/Y185F was less effective, indicating the requirement of TEY site phosphorylation. Our study suggests that functions of ERK1/2 other than its "canonical" kinase activity are also involved in the pathway-mediated growth arrest signaling. PMID- 19805546 TI - Amyotrophic lateral sclerosis-linked mutant SOD1 sequesters Hu antigen R (HuR) and TIA-1-related protein (TIAR): implications for impaired post-transcriptional regulation of vascular endothelial growth factor. AB - Down-regulation of vascular endothelial growth factor (VEGF) in the mouse leads to progressive and selective degeneration of motor neurons similar to amyotrophic lateral sclerosis (ALS). In mice expressing ALS-associated mutant superoxide dismutase 1 (SOD1), VEGF mRNA expression in the spinal cord declines significantly prior to the onset of clinical manifestations. In vitro models suggest that dysregulation of VEGF mRNA stability contributes to that decline. Here, we show that the major RNA stabilizer, Hu Antigen R (HuR), and TIA-1 related protein (TIAR) colocalize with mutant SOD1 in mouse spinal cord extracts and cultured glioma cells. The colocalization was markedly reduced or abolished by RNase treatment. Immunoanalysis of transfected cells indicated that colocalization occurred in insoluble aggregates and inclusions. RNA immunoprecipitation showed a significant loss of VEGF mRNA binding to HuR and TIAR in mutant SOD1 cells, and there was marked depletion of HuR from polysomes. Ectopic expression of HuR in mutant SOD1 cells more than doubled the mRNA half life of VEGF and significantly increased expression to that of wild-type SOD1 control. Cellular effects produced by mutant SOD1, including impaired mitochondrial function and oxidative stress-induced apoptosis, were reversed by HuR in a gene dose-dependent pattern. In summary, our findings indicate that mutant SOD1 impairs post-transcriptional regulation by sequestering key regulatory RNA-binding proteins. The rescue effect of HuR suggests that this impairment, whether related to VEGF or other potential mRNA targets, contributes to cytotoxicity in ALS. PMID- 19805547 TI - A bifunctional enzyme in a single gene catalyzes the incorporation of GlcN into the Aeromonas core lipopolysaccharide. AB - The core lipopolysaccharide (LPS) of Aeromonas hydrophila AH-3 and Aeromonas salmonicida A450 is characterized by the presence of the pentasaccharide alpha-d GlcN-(1-->7)-l-alpha-d-Hep-(1-->2)-l-alpha-d-Hep-(1-->3)-l-alpha-d-Hep-(1-->5) alpha-Kdo. Previously it has been suggested that the WahA protein is involved in the incorporation of GlcN residue to outer core LPS. The WahA protein contains two domains: a glycosyltransferase and a carbohydrate esterase. In this work we demonstrate that the independent expression of the WahA glycosyltransferase domain catalyzes the incorporation of GlcNAc from UDP-GlcNAc to the outer core LPS. Independent expression of the carbohydrate esterase domain leads to the deacetylation of the GlcNAc residue to GlcN. Thus, the WahA is the first described bifunctional glycosyltransferase enzyme involved in the biosynthesis of core LPS. By contrast in Enterobacteriaceae containing GlcN in their outer core LPS the two reactions are performed by two different enzymes. PMID- 19805548 TI - Proteomics reveals a physical and functional link between hepatocyte nuclear factor 4alpha and transcription factor IID. AB - Proteomic analyses have contributed substantially to our understanding of diverse cellular processes. Improvements in the sensitivity of mass spectrometry approaches are enabling more in-depth analyses of protein-protein networks and, in some cases, are providing surprising new insights into well established, longstanding problems. Here, we describe such a proteomic analysis that exploits MudPIT mass spectrometry and has led to the discovery of a physical and functional link between the orphan nuclear receptor hepatocyte nuclear factor 4alpha (HNF4alpha) and transcription factor IID (TFIID). A systematic characterization of the HNF4alpha-TFIID link revealed that the HNF4alpha DNA binding domain binds directly to the TATA box-binding protein (TBP) and, through this interaction, can target TBP or TFIID to promoters containing HNF4alpha binding sites in vitro. Supporting the functional significance of this interaction, an HNF4alpha mutation that blocks binding of TBP to HNF4alpha interferes with HNF4alpha transactivation activity in cells. These findings identify an unexpected role for the HNF4alpha DNA-binding domain in mediating key regulatory interactions and provide new insights into the roles of HNF4alpha and TFIID in RNA polymerase II transcription. PMID- 19805549 TI - STRA8 shuttles between nucleus and cytoplasm and displays transcriptional activity. AB - Stra8 (stimulated by retinoic acid 8) encodes a protein crucial for mammalian germ cells entering into premeiotic stages. Here, to elucidate the still unknown STRA8 molecular functions, we studied the cellular localization of the protein in several cell types, including premeiotic mouse germ cells and stem cell lines. We reported distinct STRA8 localization in germ and stem cell types and a heterogeneous protein distribution in the cytoplasm and nucleus of such cells suggesting that the protein can shuttle between these two compartments. Moreover, we identified specific protein motifs determining its nuclear import/export. Furthermore, we demonstrated that in transfected cell lines the nuclear import of STRA8 is an active process depending on an N-terminal basic nuclear localization signal. Moreover, its nuclear export is mainly mediated by the Exportin1 (XPO1) recognition of a nuclear export signal. Significantly, we also demonstrated that STRA8 associates with DNA and possesses transcriptional activity. These observations strongly suggest that STRA8 can exert important functions in the nucleus rather than in the cytoplasm as believed previously, likely depending on the cell type and regulated by its nuclear-cytoplasmic shuttling. PMID- 19805550 TI - Metal-free superoxide dismutase-1 and three different amyotrophic lateral sclerosis variants share a similar partially unfolded beta-barrel at physiological temperature. AB - The structure and unfolding of metal-free (apo) human wild-type SOD1 and three pathogenic variants of SOD1 (A4V, G93R, and H48Q) that cause familial amyotrophic lateral sclerosis have been studied with amide hydrogen/deuterium exchange and mass spectrometry. The results indicate that a significant proportion of each of these proteins exists in solution in a conformation in which some strands of the beta-barrel (i.e. beta2) are well protected from exchange at physiological temperature (37 degrees C), whereas other strands (i.e. beta3 and beta4) appear to be unprotected from hydrogen/deuterium exchange. Moreover, the thermal unfolding of these proteins does not result in the uniform incorporation of deuterium throughout the polypeptide but involves the local unfolding of different residues at different temperatures. Some regions of the proteins (i.e. the "Greek key" loop, residues 104-116) unfold at a significantly higher temperature than other regions (i.e. beta3 and beta4, residues 21-53). Together, these results show that human wild-type apo-SOD1 and variants have a partially unfolded beta-barrel at physiological temperature and unfold non-cooperatively. PMID- 19805551 TI - Reproductive aging is associated with altered gene expression in human luteinized granulosa cells. AB - Declining reproductive success with aging is attributable to qualitative and quantitative deterioration in oocytes, which are nurtured by granulosa cells (GCs). This prospective study assesses whether reproductive aging is accompanied by differential gene expression in luteinized GCs from in vitro fertilization (IVF) patients. Women with nonovarian infertility etiologies were categorized as younger (< or =30, n = 3) or older (> or =40, n = 3). During oocyte retrieval, mural GCs were isolated; messenger RNA (mRNA) was extracted and transcribed for complementary DNA (cDNA) microarray analysis. Differential gene expression was confirmed by real-time polymerase chain reaction (PCR). Analysis revealed 120 genes were differentially expressed. Three genes were upregulated and 117 were downregulated (including interleukin [IL]-1beta, IL-1R2, and IL-6R) in GCs of older versus younger patients. Our data provide evidence of downregulation in IL 1 and IL-6 gene families in luteinized GCs with advancing age. Given previously recognized roles for the IL gene family in folliculogenesis and ovulation, our findings may partly explain ovulatory and luteal dysfunctions associated with reproductive aging. PMID- 19805552 TI - Uterine leiomyomas exhibit fewer stem/progenitor cell characteristics when compared with corresponding normal myometrium. AB - Uterine leiomyomas (also known as uterine fibroids) are the most common benign tumors of female reproductive tract and are the single most common indication for hysterectomies. Despite their high prevalence, the exact pathogenesis of these benign tumors is still unknown. One possible mechanism for leiomyoma formation is dysregulation of mesenchymal stem cell activity. Mesenchymal stem cells have been identified in both human and murine uteri and cancer stem cells have been identified in female reproductive malignancies. We compared stem/progenitor cell characteristics in both normal myometrium and the corresponding leiomyoma of patient's undergoing hysterectomies. We found that leiomyoma cells form fewer mesenchymal stem cell colonies and exhibit less Hoechst dye-excluding side population (SP) activity, which is a function associated with progenitor cells in other tissues, than cells isolated from normal myometrium. Whereas in normal myometrium, we observed heterogeneous expression of CD90, a cell surface marker associated the with differentiation potential of uterine fibroblasts, in leiomyomas, we observed homogenous expression of CD90, suggesting leiomyoma cells are more terminally differentiated. Furthermore, we found that while leiomyoma cells could only produce CD90 expressing cells, both CD90+ and CD90- myometrial cells could reestablish their original heterogeneous CD90 profile when expanded in vitro. These results suggest that normal myometrium contains cells with stem/progenitor cell activities that are absent in leiomyomas. PMID- 19805553 TI - Global challenge of antibiotic-resistant Treponema pallidum. AB - Syphilis is a multistage infectious disease that is usually transmitted through contact with active lesions of a sexual partner or from an infected pregnant woman to her fetus. Despite elimination efforts, syphilis remains endemic in many developing countries and has reemerged in several developed countries, including China, where a widespread epidemic recently occurred. In the absence of a vaccine, syphilis control is largely dependent upon identification of infected individuals and treatment of these individuals and their contacts with antibiotics. Although penicillin is still effective, clinically significant resistance to macrolides, a second-line alternative to penicillin, has emerged. Macrolide-resistant strains of Treponema pallidum are now prevalent in several developed countries. An understanding of the genetic basis of T. pallidum antibiotic resistance is essential to enable molecular surveillance. This review discusses the genetic basis of T. pallidum macrolide resistance and the potential of this spirochete to develop additional antibiotic resistance that could seriously compromise syphilis treatment and control. PMID- 19805554 TI - In vitro pharmacodynamics of levofloxacin and other aerosolized antibiotics under multiple conditions relevant to chronic pulmonary infection in cystic fibrosis. AB - The inhalational administration of antibiotics can provide high concentrations locally in the lungs of cystic fibrosis patients and, thus, can be useful for the treatment of chronic bacterial infections. The present study evaluated the in vitro activities of levofloxacin, ciprofloxacin, tobramycin, amikacin, and aztreonam against clinical isolates of Pseudomonas aeruginosa, Burkholderia cepacia complex, Stenotrophomonas maltophilia, Alcaligenes xylosoxidans, and Staphylococcus aureus from cystic fibrosis patients. Levofloxacin was the most potent antibiotic against all cystic fibrosis isolates tested, with MIC(90)s ranging from 8 to 32 microg/ml. Levofloxacin was more potent than the aminoglycosides and aztreonam against P. aeruginosa biofilms. Time-kill assays with drug concentrations achievable in sputum following aerosol administration showed that levofloxacin had the most rapid rate of killing among mucoid and nonmucoid isolates of P. aeruginosa. In contrast to tobramycin, the bactericidal activity of levofloxacin was not affected by sputum from cystic fibrosis patients. The results of the study show that the high concentrations of levofloxacin readily achievable in the lung following aerosol delivery may be useful for the management of pulmonary infections in patients with cystic fibrosis. PMID- 19805555 TI - Mitochondrial DNA depletion and respiratory chain activity in primary human subcutaneous adipocytes treated with nucleoside analogue reverse transcriptase inhibitors. AB - Mitochondrial dysfunction as a consequence of mitochondrial DNA (mtDNA) depletion due to therapy with nucleoside analogue reverse transcriptase inhibitors (NRTI) has been proposed as a pathogenic mechanism leading to lipoatrophy in HIV infected patients. The aim of our study was to investigate the impact of NRTI treatment on mtDNA abundance and the activities of respiratory chain complexes in primary human subcutaneous preadipocytes (phsPA). We studied adipocyte phenotypes, viability, and differentiation (CCAAT/enhancer-binding protein alpha [C/EBPalpha] and peroxisome proliferator-activated receptor gamma [PPARgamma] expression) and adiponectin production, mtDNA content, mitochondrial membrane potential, mitochondrial mass, and respiratory chain enzyme and citrate synthase activities in both proliferating and differentiating phsPA. Cells were exposed to zidovudine (6 microM), stavudine (d4T; 3 microM), and zalcitabine (ddC; 0.1 microM) for 8 weeks. NRTI-induced mtDNA depletion occurred in proliferating and differentiating phsPA after exposure to therapeutic drug concentrations of d4T and ddC. At these concentrations, ddC and d4T led to an almost 50% decrease in the number of mtDNA copies per cell without major impact on adipocyte differentiation. Despite mtDNA depletion by NRTI, the activities of the respiratory chain complexes, the mitochondrial membrane potential, and the mitochondrial mass were found to be unaffected. Severe NRTI-mediated mtDNA depletion in phsPA is not inevitably associated with impaired respiratory chain activity or altered mitochondrial membrane potential. PMID- 19805556 TI - Metronidazole resistance in Prevotella spp. and description of a new nim gene in Prevotella baroniae. AB - Nonduplicate clinical isolates of Prevotella spp. recovered from patients hospitalized between 2003 and 2006 in two French tertiary-care teaching hospitals were investigated for their susceptibility to metronidazole and the presence of nim genes. Of the 188 strains tested, 3 isolates displayed reduced susceptibility to metronidazole after 48 h of incubation, while 27 additional isolates exhibited heterogeneous resistance after prolonged incubation; all 30 of the isolates were nim negative. Among the remaining 158 isolates, 7 nim-positive isolates were detected. All of these strains were identified as Prevotella baroniae by 16S rRNA gene sequence analysis and contained a new nim gene, named nimI, as determined by DNA sequence analysis. Chromosomal localization of this single-copy gene was demonstrated in all clinical isolates as well as in type strain P. baroniae DSM 16972 by using Southern hybridization. No known associated insertion sequence elements were detected upstream of the nimI gene in any of the nim-positive strains by PCR mapping. After prolonged exposure to metronidazole, stable resistant subpopulations could be selected in nimI-positive Prevotella isolates (n = 6) as well as in nim-negative Prevotella isolates (n = 6), irrespective of their initial susceptibility to this antibiotic. This study is the first description of a new nitroimidazole resistance gene in P. baroniae which seems to be silent and which might be intrinsic in this species. Moreover, our findings highlight the fact that high-level resistance to metronidazole may be easily induced in both nim-positive and nim-negative Prevotella sp. strains. PMID- 19805557 TI - Mutations in ribosomal protein L3 are associated with oxazolidinone resistance in staphylococci of clinical origin. AB - Following recent reports of ribosomal protein L3 mutations in laboratory-derived linezolid-resistant (LZD(r)) Staphylococcus aureus, we investigated whether similar mutations were present in LZD(r) staphylococci of clinical origin. Sequence analysis of a variety of LZD(r) isolates revealed two L3 mutations, DeltaSer145 (S. aureus NRS127) and Ala157Arg (Staphylococcus epidermidis 1653059), both occurring proximal to the oxazolidinone binding site in the peptidyl transferase center. The oxazolidinone torezolid maintained a >or=8-fold potency advantage over linezolid for both strains. PMID- 19805558 TI - Correlation between vancomycin MIC values and those of other agents against gram positive bacteria among patients with bloodstream infections caused by methicillin-resistant Staphylococcus aureus. AB - An increase in the distribution of vancomycin MIC values among methicillin (meticillin)-resistant Staphylococcus aureus (MRSA) isolates has been noted. It is postulated that the shift in vancomycin MIC values may be associated with a concurrent rise in the MIC values of other anti-MRSA agents. Scant data are available on the correlation between vancomycin MIC values and the MIC values of other anti-MRSA agents. This study examined the correlation between vancomycin MIC values and the MIC values of daptomycin, linezolid, tigecycline, and teicoplanin among 120 patients with bloodstream infections caused by MRSA at a tertiary care hospital between January 2005 and May 2007. For each included patient, the MIC values of the antibiotics under study were determined by the Etest method and were separated into the following two categories: day 1 (index) and post-day 1 (subsequent). For subsequent isolates, the MIC values for each antibiotic from the post-day 1 terminal isolate were used. Among the index isolates, there was a significant correlation (P value, <0.01) between the MIC values for vancomycin and daptomycin and between the MIC values for vancomycin and teicoplanin. The MIC values for daptomycin were significantly correlated with linezolid, tigecycline, and teicoplanin MIC values. Among the 48 patients with subsequent isolates, vancomycin MIC values were significantly correlated with MIC values for daptomycin, linezolid, and teicoplanin (rho value of >or=0.38 for all comparisons). This study documented an association between vancomycin MIC values and the MIC values of other anti-MRSA antibiotics among patients with bloodstream infections caused by MRSA primarily treated with vancomycin. PMID- 19805559 TI - Prevalence and clonal characterization of Streptococcus pyogenes clinical isolates with reduced fluoroquinolone susceptibility in Spain. AB - The aim of this study was to determine the prevalence and characteristics of non fluoroquinolone (FQ)-susceptible Streptococcus pyogenes isolates and to study their mechanisms of resistance. We performed a prospective prevalence study with 468 isolates collected from 2005 to 2007 and a retrospective study that was based on the examination of existing data collected from 1999 to 2008. The retrospective study included data for isolates with high-level resistance (HR) to ciprofloxacin (MIC >or= 32 microg/ml) (HR isolates) and isolates with the same emm types as those reported in the literature with low-level resistance (LR) to ciprofloxacin (MICs, 2 to 8 microg/ml) (LR isolates, n = 205). Genetic characterization of the isolates was performed by means of emm typing and multilocus sequence typing. The prevalence of LR ranged from 1.9% in 2005 to 30.8% in 2007. This increase was mainly due to the circulation of an emm6 subtype (emm6.4) that represented 77.1% of the LR isolates in 2007. Notably, another emm6 subtype, also detected in 2007 (emm6.37), showed coresistance to 14- and 15 membered macrolides mediated by the mefA gene. Only three HR isolates were detected (isolates emm68.1/ST247/T3,13,B3264, emm77/ST399/T28, and emm28/ST52/T28), and all were identified in the retrospective study. Overall, the 673 isolates represented 25 emm types. All LR isolates were clustered into two emm types: emm6 (six emm6 subtypes) and emm75. All the 156 emm6 isolates had LR, harbored the Ser79/Ala mutation in the parC gene product, and had the same sequence type (ST), ST382. Most (21/33) of the emm75 isolates had LR, showed the Ser79/Phe plus Asp91/Asn double mutation in the parC gene product, and were ST150. The Asp91/Asn mutation by itself did not confer resistance to FQs. PMID- 19805560 TI - In vitro activity of gallium maltolate against Staphylococci in logarithmic, stationary, and biofilm growth phases: comparison of conventional and calorimetric susceptibility testing methods. AB - Ga(3+) is a semimetal element that competes for the iron-binding sites of transporters and enzymes. We investigated the activity of gallium maltolate (GaM), an organic gallium salt with high solubility, against laboratory and clinical strains of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), methicillin-susceptible Staphylococcus epidermidis (MSSE), and methicillin-resistant S. epidermidis (MRSE) in logarithmic or stationary phase and in biofilms. The MICs of GaM were higher for S. aureus (375 to 2000 microg/ml) than S. epidermidis (94 to 200 microg/ml). Minimal biofilm inhibitory concentrations were 3,000 to >or=6,000 microg/ml (S. aureus) and 94 to 3,000 microg/ml (S. epidermidis). In time-kill studies, GaM exhibited a slow and dose-dependent killing, with maximal action at 24 h against S. aureus of 1.9 log(10) CFU/ml (MSSA) and 3.3 log(10) CFU/ml (MRSA) at 3x MIC and 2.9 log(10) CFU/ml (MSSE) and 4.0 log(10) CFU/ml (MRSE) against S. epidermidis at 10x MIC. In calorimetric studies, growth-related heat production was inhibited by GaM at subinhibitory concentrations; and the minimal heat inhibition concentrations were 188 to 4,500 microg/ml (MSSA), 94 to 1,500 microg/ml (MRSA), and 94 to 375 microg/ml (MSSE and MRSE), which correlated well with the MICs. Thus, calorimetry was a fast, accurate, and simple method useful for investigation of antimicrobial activity at subinhibitory concentrations. In conclusion, GaM exhibited activity against staphylococci in different growth phases, including in stationary phase and biofilms, but high concentrations were required. These data support the potential topical use of GaM, including its use for the treatment of wound infections, MRSA decolonization, and coating of implants. PMID- 19805561 TI - Novel approach for comparing the abilities of quinolones to restrict the emergence of resistant mutants during quinolone exposure. AB - An agar-plate assay was adapted to examine aspects of quinolone structure that restrict the emergence of quinolone-mediated quinolone resistance. When Escherichia coli was applied to agar containing nalidixic acid, the number of quinolone-resistant mutants arising during incubation was decreased by raising the drug concentration and by mutations expected to block the induction of the SOS response (recA, lexA); the mutant number was increased by a mutator mutation (ung). The examination of four related fluoroquinolones then revealed that a C-8 methoxy group and an N-ethyl piperazine substituent at C-7 reduced mutant acquisition more effectively than C-8 H and C-7 C-ethyl piperazine groups. The fluoroquinolone that was most effective at restricting mutant acquisition was the most active when lethal activity was measured on agar plates or in liquid medium (as minimal bactericidal concentration). It also exhibited the lowest ratio of mutant MIC to wild-type MIC when it was tested with a set of isogenic gyrase mutants, and it had a low mutant prevention concentration (MPC) relative to MIC. However, a low MPC was less likely to be important in restricting the induced mutant accumulation because a fluoroquinolone N-ethyl piperazine substituent was more effective than a C-ethyl piperazine substituent at reducing mutant accumulation but was less effective at lowering the MPC. An 8-methoxy-quinazoline 2,4-dione was also effective at restricting the accumulation of resistant mutants on agar. Collectively, these data characterize a simple assay for detection of drug-mediated resistance that is sensitive to the structures of GyrA inhibitors. The assay provides a new method for screening quinolones and quinolone-like molecules that complements MPC-based tests for restricting the emergence of resistance. PMID- 19805562 TI - Novel antiseptic urinary catheters for prevention of urinary tract infections: correlation of in vivo and in vitro test results. AB - Urinary catheters are widely used for hospitalized patients and are often associated with high rates of urinary tract infection. We evaluated in vitro the antiadherence activity of a novel antiseptic Gendine-coated urinary catheter against several multidrug-resistant bacteria. Gendine-coated urinary catheters were compared to silver hydrogel-coated Foley catheters and uncoated catheters. Bacterial biofilm formation was assessed by quantitative culture and scanning electron microscopy. These data were further correlated to an in vivo rabbit model. We challenged 31 rabbits daily for 4 days by inoculating the urethral meatus with 1.0 x 10(9) CFU streptomycin-resistant Escherichia coli per day. In vitro, Gendine-coated urinary catheters reduced the CFU of all organisms tested for biofilm adherence compared with uncoated and silver hydrogel-coated catheters (P < 0.004). Scanning electron microscopy analysis showed that a thick biofilm overlaid the control catheter and the silver hydrogel-coated catheters but not the Gendine-coated urinary catheter. Similar results were found with the rabbit model. Bacteriuria was present in 60% of rabbits with uncoated catheters and 71% of those with silver hydrogel-coated catheters (P < 0.01) but not in those with Gendine-coated urinary catheters. No rabbits with Gendine-coated urinary catheters had invasive bladder infections. Histopathologic assessment revealed no differences in toxicity or staining. Gendine-coated urinary catheters were more efficacious in preventing catheter-associated colonization and urinary tract infections than were silver hydrogel-coated Foley catheters and uncoated catheters. PMID- 19805563 TI - Amiodarone and miltefosine act synergistically against Leishmania mexicana and can induce parasitological cure in a murine model of cutaneous leishmaniasis. AB - Leishmaniasis is parasitic disease that is an important problem of public health worldwide. Intramuscularly administered glucantime and pentostam are the most common drugs used for treatment of this disease, but they have significant limitations due to toxicity and increasing resistance. A recent breakthrough has been the introduction of orally administered miltefosine for the treatment of visceral, cutaneous, and mucocutaneous leishmaniasis, but the relative high cost and concerns about teratogenicity have limited the use of this drug. Searching for alternative drugs, we previously demonstrated that the antiarrhythmic drug amiodarone is active against Leishmania mexicana promastigotes and intracellular amastigotes, acting via disruption of intracellular Ca(2+) homeostasis (specifically at the mitochondrion and the acidocalcisomes of these parasites) and through inhibition of the parasite's de novo sterol biosynthesis (X. Serrano Martin, Y. Garcia-Marchan, A. Fernandez, N. Rodriguez, H. Rojas, G. Visbal, and G. Benaim, Antimicrob. Agents Chemother. 53:1403-1410, 2009). In the present work, we found that miltefosine also disrupts the parasite's intracellular Ca(2+) homeostasis, in this case by inducing a large increase in intracellular Ca(2+) levels, probably through the activation of a plasma membrane Ca(2+) channel. We also investigated the in vitro and in vivo activities of amiodarone and miltefosine, used alone or in combination, on L. mexicana. It was found that the drug combination had synergistic effects on the proliferation of intracellular amastigotes growing inside macrophages and led 90% of parasitological cures in a murine model of leishmaniasis, as revealed by a PCR assay using a novel DNA sequence specific for L. mexicana. PMID- 19805564 TI - Prevention and treatment of Staphylococcus aureus pneumonia with a beta cyclodextrin derivative. AB - Staphylococcus aureus pneumonia is a common, potentially life-threatening infection caused by this human pathogen. The only therapies available to treat S. aureus pneumonia are antibiotics, a modality that is jeopardized by the organism's remarkable ability to acquire antimicrobial resistance. S. aureus alpha-hemolysin is a pore-forming cytotoxin that is essential for the pathogenesis of pneumonia. Strains lacking this cytotoxin are avirulent in a murine model of pneumonia; likewise, vaccine-based strategies that antagonize the toxin afford protection against lethal disease. Disruption of the function of this toxin therefore provides a potent mechanism to prevent and/or treat S. aureus pneumonia. beta-Cyclodextrin derivatives are small molecules with a sevenfold symmetry that mirrors the heptameric alpha-hemolysin. These compounds block the assembled alpha-hemolysin pore, compromising toxin function. We report that a modified beta-cyclodextrin compound, IB201, prevents alpha-hemolysin induced lysis of human alveolar epithelial cells. This protective effect does not result from the ability of the beta-cyclodextrin to impair formation of the oligomeric alpha-hemolysin on the cell surface, supporting a role for this molecule in blockade of the lytic pore. An examination of IB201 in murine S. aureus pneumonia demonstrated that administration of this compound prevents and treats disease, protecting against mortality. Consistent with the vital importance of alpha-hemolysin in pneumonia caused by methicillin-sensitive and highly virulent methicillin-resistant S. aureus strains, IB201 protects against lethal challenge with both types of isolates. These observations, coupled with a favorable safety profile of beta-cyclodextrin compounds, provide a novel strategy that may be developed to combat S. aureus pneumonia. PMID- 19805565 TI - Phenotypic and genotypic characterization of Enterobacteriaceae with decreased susceptibility to carbapenems: results from large hospital-based surveillance studies in China. AB - The resistance mechanism of 49 Enterobacteriaceae isolates with decreased susceptibility to carbapenems collected from 2004 to 2008 at 16 teaching hospitals in China was investigated. Moderate- to high-level carbapenem resistance in most isolates was more closely associated with loss or decreased expression of both major porins combined with production of AmpC or extended spectrum beta-lactamase enzymes, while KPC-2, IMP-4, and IMP-8 carbapenemase production may lead to a low to moderate level of carbapenem resistance in Enterobacteriaceae in China. PMID- 19805566 TI - Clindamycin-induced CovS-mediated regulation of the production of virulent exoproteins streptolysin O, NAD glycohydrolase, and streptokinase in Streptococcus pyogenes. AB - The administration of high-dose clindamycin (CLI) along with penicillin is recommended for the treatment of streptococcal toxic shock syndrome. However, the prevalence of CLI-resistant Streptococcus pyogenes strains is increasing worldwide, and the effect of CLI on CLI-resistant S. pyogenes strains remains unknown. We aimed to evaluate the effect of CLI on the in vitro production of three major virulent exoproteins, namely, streptolysin O (Slo), NAD glycohydrolase (Nga), and streptokinase (Ska), by CLI-resistant S. pyogenes strains. After the incubation of M1 serotype CLI-resistant S. pyogenes D2TY in the presence of 1 microg/ml CLI, the amounts of Slo, Nga, and Ska and the levels of slo, nga, and ska mRNA in the supernatant were analyzed by Northern blotting and Western blotting, respectively. The results of both assays showed that the production of Slo, Nga, and Ska was higher with CLI treatment than without CLI treatment. We evaluated the role of the sensor kinase CovS, which is involved in the two-component system of S. pyogenes, in the CLI-induced production of these three exoproteins. Northern blotting analysis revealed that CLI induced the expression of covS mRNA in wild-type strain D2TY. Furthermore, both Northern blotting and Western blotting analyses showed that CLI decreased the levels of expression of Slo, Nga, and Ska in isogenic covS mutant D2TYcovS. These results suggest that CLI increases the production of three virulent exoproteins in CLI resistant S. pyogenes strains via the action of CovS. PMID- 19805567 TI - Long-lasting enfuvirtide carrier pentasaccharide conjugates with potent anti human immunodeficiency virus type 1 activity. AB - Enfuvirtide (also known as Fuzeon, T-20, or DP-178) is an antiretroviral fusion inhibitor which prevents human immunodeficiency virus type 1 (HIV-1) from entering host cells. This linear 36-mer synthetic peptide is indicated, in combination with other antiretroviral agents, for the treatment of HIV-1-infected individuals and AIDS patients with multidrug-resistant HIV infections. Although enfuvirtide is an efficient anti-HIV-1 drug, its clinical use is limited by a short plasma half-life, i.e., approximately 2 h, which requires twice-daily subcutaneous injections, often resulting in skin sensitivity reaction side effects at the injection sites. Ultimately, 80% of patients stop enfuvirtide treatment within 6 months because of these side effects. We report on the development of long-lasting enfuvirtide conjugates by the use of the site specific conjugation of enfuvirtide to an antithrombin-binding carrier pentasaccharide (CP) through polyethylene glycol (PEG) linkers of various lengths. These conjugates showed consistent and broad anti-HIV-1 activity in the nanomolar range. The coupling of the CP to enfuvirtide only moderately affected the in vitro anti-HIV-1 activity in the presence of antithrombin. Most importantly, one of these conjugates, enfuvirtide-PEG(12)-CP (EP40111), exhibited a prolonged elimination half-life of more than 10 h in rat plasma compared to the half-life of native enfuvirtide, which was 2.8 h. On the basis of the pharmacokinetic properties of antithrombin-binding pentasaccharides, the anticipated half-life of EP40111 in humans would putatively be about 120 h, which would allow subcutaneous injection once a week instead of twice daily. In conclusion, EP40111 is a promising compound with strong potency as a novel long lasting anti-HIV-1 drug. PMID- 19805568 TI - Eradication of intracellular Salmonella enterica serovar Typhimurium with a small molecule, host cell-directed agent. AB - Eradication of intracellular pathogenic bacteria with host-directed chemical agents has been an anticipated innovation in the treatment of antibiotic resistant bacteria. We previously synthesized and characterized a novel small molecule agent, AR-12, that induces autophagy and inhibits the Akt kinase in cancer cells. As both autophagy and the Akt kinase have been shown recently to play roles in the intracellular survival of several intracellular bacteria, including Salmonella enterica serovar Typhimurium, we investigated the effect of AR-12 on the intracellular survival of Salmonella serovar Typhimurium in macrophages. Our results show that AR-12 induces autophagy in macrophages, as indicated by increased autophagosome formation, and potently inhibits the survival of serovar Typhimurium in macrophages in association with increased colocalization of intracellular bacteria with autophagosomes. Intracellular bacterial growth was partially rescued in the presence of AR-12 by the short hairpin RNA-mediated knockdown of Beclin-1 or Atg7 in macrophages. Moreover, AR 12 inhibits Akt kinase activity in infected macrophages, which we show to be important for its antibacterial effect as the enforced expression of constitutively activated Akt1 in these cells reverses the AR-12-induced inhibition of intracellular serovar Typhimurium survival. Finally, oral administration of AR-12 at 2.5 mg/kg/day to serovar Typhimurium-infected mice reduced hepatic and splenic bacterial burdens and significantly prolonged survival. These findings show that AR-12 represents a proof of principle that the survival of intracellular bacteria can be suppressed by small-molecule agents that target both innate immunity and host cell factors modulated by bacteria. PMID- 19805569 TI - Bacterial strain-to-strain variation in pharmacodynamic index magnitude, a hitherto unconsidered factor in establishing antibiotic clinical breakpoints. AB - Antibiotic pharmacodynamic modeling allows variations in pathogen susceptibility and human pharmacokinetics to be accounted for when considering antibiotic doses, potential bacterial pathogen targets for therapy, and clinical susceptibility breakpoints. Variation in the pharmacodynamic index (area-under-the-concentration curve to 24 h [AUC(24)]/MIC; maximum serum concentration of drug in the serum/MIC; time the serum concentration remains higher than the MIC [T > MIC]) is not usually considered. In an in vitro pharmacokinetic model of infection using a dose-ranging design, we established the relationship between AUC(24)/MIC and the antibacterial effect for moxifloxacin against 10 strains of Staphylococcus aureus. The distributions of AUC(24)/MIC targets for 24-h bacteriostatic effect and 1-log, 2-log, and 3-log drops in bacterial counts were used to calculate potential clinical breakpoint values, and these were compared with those obtained by the more conventional approach of taking a single AUC(24)/MIC target. Consideration of the AUC(24)/MIC as a distribution rather than a single value resulted in a lower clinical breakpoint. PMID- 19805570 TI - Polyamine transport as a target for treatment of Pneumocystis pneumonia. AB - Polyamine levels are greatly increased in alveolar macrophages (AMs) during Pneumocystis pneumonia (PCP), leading to increased production of H(2)O(2), which causes AMs to undergo apoptosis. One of the mechanisms by which polyamine levels in AMs are elevated is enhanced uptake of exogenous polyamines. In this study, the possibility of targeting polyamine uptake as a treatment for PCP was examined. Four anthracene- and one benzene-polyamine conjugates that are potential polyamine transport inhibitors, including N1-anthracen-9-ylmethyl butane-1,4-diamine; N-(4-aminobutyl)-N-anthracen-9-ylmethylbutane-1,4-diamine; N [4-(4-aminobutylamino)butyl]-N-anthracen-9-ylmethylbutane-1,4-diamine; N-(4-amino butyl)-N'-(10-[[4-(4-amino-butylamino)butylamino]-methyl]anthracen-9 ylmethyl)butane-1,4-diamine (44-Ant-44); and benzene-polyamine conjugate N-(4 amino-butyl)-N'-(4-[[4-(4-amino-butylamino)butylamino]-methyl]benzyl)butane-1,4 diamine (44-Bn-44), were tested. Compounds 44-Ant-44 and 44-Bn-44 were found to have a very low toxicity to AMs in vitro and were evaluated for their therapeutic effect on PCP in vivo. Sprague-Dawley rats infected with P. carinii for 28 days were intranasally instilled with 50 microl of a 1 mM solution of 44-Bn-44 or 44 Ant-44 every 2 days. Twenty-one days after initiation of the treatment, three to five rats from each group were sacrificed and examined for lung pathology, organism burden, and apoptosis of AMs. Both 44-Bn-44 and 44-Ant-44 reduced organism burdens; however, only 44-Ant-44 decreased the severity of the infection with reduced lung inflammation, increased clearance of exudates, increased air space, and decreased apoptosis of AMs. 44-Ant-44 also significantly prolonged the survival of treated animals. These results suggest that polyamine uptake is a potential target for treatment of PCP. PMID- 19805571 TI - New small-molecule inhibitor class targeting human immunodeficiency virus type 1 virion maturation. AB - A new small-molecule inhibitor class that targets virion maturation was identified from a human immunodeficiency virus type 1 (HIV-1) antiviral screen. PF-46396, a representative molecule, exhibits antiviral activity against HIV-1 laboratory strains and clinical isolates in T-cell lines and peripheral blood mononuclear cells (PBMCs). PF-46396 specifically inhibits the processing of capsid (CA)/spacer peptide 1 (SP1) (p25), resulting in the accumulation of CA/SP1 (p25) precursor proteins and blocked maturation of the viral core particle. Viral variants resistant to PF-46396 contain a single amino acid substitution in HIV-1 CA sequences (CAI201V), distal to the CA/SP1 cleavage site in the primary structure, which we demonstrate is sufficient to confer significant resistance to PF-46396 and 3-O-(3',3'-dimethylsuccinyl) betulinic acid (DSB), a previously described maturation inhibitor. Conversely, a single amino substitution in SP1 (SP1A1V), which was previously associated with DSB in vitro resistance, was sufficient to confer resistance to DSB and PF-46396. Further, the CAI201V substitution restored CA/SP1 processing in HIV-1-infected cells treated with PF 46396 or DSB. Our results demonstrate that PF-46396 acts through a mechanism that is similar to DSB to inhibit the maturation of HIV-1 virions. To our knowledge, PF-46396 represents the first small-molecule HIV-1 maturation inhibitor that is distinct in chemical class from betulinic acid-derived maturation inhibitors (e.g., DSB), demonstrating that molecules of diverse chemical classes can inhibit this mechanism. PMID- 19805572 TI - Effects of subinhibitory concentrations of antibiotics on colonization factor expression by moxifloxacin-susceptible and moxifloxacin-resistant Clostridium difficile strains. AB - Recent outbreaks of Clostridium difficile infection have been related to the emergence of the NAP1/027 epidemic strain. This strain demonstrates increased virulence and resistance to the C-8-methoxyfluoroquinolones gatifloxacin and moxifloxacin. These antibiotics have been implicated as major C. difficile infection-inducing agents. We investigated by real-time reverse transcription-PCR the impact of subinhibitory concentrations of ampicillin, clindamycin, ofloxacin, and moxifloxacin on the expression of genes encoding three colonization factors, the protease Cwp84, the high-molecular-weight S-layer protein, and the fibronectin-binding protein Fbp68. We have previously shown in six non-NAP1/027 moxifloxacin-susceptible strains that the presence of ampicillin or clindamycin induced an upregulation of these genes, whereas the presence of fluoroquinolones did not. The objective of this study was to analyze the expression of these genes under the same conditions in four NAP1/027 strains, one moxifloxacin susceptible and three moxifloxacin resistant. Two in vitro-selected moxifloxacin-resistant mutants were also analyzed. Moxifloxacin resistance was associated with the Thr82 ->Ile substitution in GyrA in all but one of the moxifloxacin-resistant strains. The expression of cwp84 and slpA was strongly increased after culture with ampicillin or clindamycin in NAP1/027 strains. Interestingly, after culture with fluoroquinolones, the expression of cwp84 and slpA was only increased in four moxifloxacin-resistant strains, including the NAP1/027 strains and one of the in vitro-selected mutants. The overexpression of cwp84 was correlated with increased production of the protease Cwp84. The historical NAP1/027 moxifloxacin susceptible strain and its mutant appear to be differently regulated by fluoroquinolones. Overall, fluoroquinolones appear to favor the expression of some colonization factor-encoding genes in resistant C. difficile strains. The fluoroquinolone resistance of the NAP1/027 epidemic strains could be considered an ecological advantage. This could also increase their colonization fitness and promote the infection. PMID- 19805573 TI - Transcriptomic profiling of the Saccharomyces cerevisiae response to quinine reveals a glucose limitation response attributable to drug-induced inhibition of glucose uptake. AB - Quinine has been employed in the treatment of malaria for centuries and is still used against severe Plasmodium falciparum malaria. However, its interactions with the parasite remain poorly understood and subject to debate. In this study, we used the Saccharomyces cerevisiae eukaryotic model to better understand quinine's mode of action and the mechanisms underlying the cell response to the drug. We obtained a transcriptomic profile of the yeast's early response to quinine, evidencing a marked activation of genes involved in the low-glucose response (e.g., CAT8, ADR1, MAL33, MTH1, and SNF3). We used a low inhibitory quinine concentration with no detectable effect on plasma membrane function, consistent with the absence of a general nutrient starvation response and suggesting that quinine-induced glucose limitation is a specific response. We have further shown that transport of [(14)C]glucose is inhibited by quinine, with kinetic data indicating competitive inhibition. Also, tested mutant strains deleted for genes encoding high- and low-affinity hexose transporters (HXT1 to HXT5, HXT8, and HXT10) exhibit resistance phenotypes, correlating with reduced levels of quinine accumulation in the mutants examined. These results suggest that the hexose transporters are facilitators of quinine uptake in S. cerevisiae, possibly through a competitive inhibition mechanism. Interestingly, P. falciparum is highly dependent on glucose uptake, which is mediated by the single-copy transporter PfHT1, a protein with high homology to yeast's hexose transporters. We propose that PfHT1 is an interesting candidate quinine target possibly involved in quinine import in P. falciparum, an uptake mechanism postulated in recent studies to occur through a still-unidentified importer(s). PMID- 19805574 TI - ELR+ CXC chemokines and oncogenic Ras-mediated tumorigenesis. AB - The small GTPase Ras is mutated to remain in the active oncogenic state in one third of human cancers, thereby promoting tumorigenesis. It has recently come to light that one consequence of oncogenic Ras signaling is secretion of cytokines vascular endothelial growth factor (VEGF), interleukin 6 (IL6), hCXCL1 (Gro alpha) and hCXCL8 (IL8). As the latter two belong to the ELR+ Cys-X-Cys (CXC) chemokine family, we investigated whether the entire family of ELR+ CXC chemokines plays a role in oncogenic Ras-mediated tumorigenesis. We now demonstrate that oncogenic Ras induced the expression and secretion of the ELR+ CXC chemokine family in different tumorigenic human cells and that these chemokines are elevated in tumor specimens. Moreover, genetic ablation of the common receptor for these chemokines, mCXCR2, reduced oncogenic Ras-driven tumorigenesis in mice. Taken together, we suggest that oncogenic Ras induces the secretion of the ELR+ CXC chemokine family to promote tumorigenesis. This chemokine signature may identify the presence of Ras activation in cancer and perhaps even serve as targets for oncogenic Ras-driven tumor cells. PMID- 19805575 TI - A case-control and a family-based association study revealing an association between CYP2E1 polymorphisms and nasopharyngeal carcinoma risk in Cantonese. AB - Nasopharyngeal carcinoma (NPC) is rare in most parts of the world but is more prevalent in Southern China, especially in Guangdong. The cytochrome P450 2E1 (CYP2E1) has been recognized as one of the critically important enzymes involved in oxidizing carcinogens and is probably to be associated with NPC carcinogenesis. To systematically investigate the association between genetic variants in CYP2E1 and NPC risk in Cantonese, two independent studies, a family based association study and a case-control study, were conducted using the haplotype-tagging single-nucleotide polymorphism approach. A total of 2499 individuals from 546 nuclear families were initially genotyped for the family based association study. Single-nucleotide polymorphisms (SNPs) rs9418990, rs915908, rs8192780, rs1536826, rs3827688 and one haplotype h2 (CGTGTTAA) were revealed to be significantly associated with the NPC phenotype (P = 0.045-0.003 and P = 0.003, respectively). To follow up the initial study, a case-control study including 755 cases and 755 controls was conducted. Similar results were observed in the case-control study in individuals <46 years of age and had a history of cigarette smoking, with odds ratios (ORs) of specific genotypes ranging from 1.88 to 2.99 corresponding to SNP rs9418990, rs3813865, rs915906, rs2249695, rs8192780, rs1536826, rs3827688 and of haplotypes h2 with OR = 1.65 (P = 0.026), h5 (CCCGTTAA) with OR = 2.58 (P = 0.007). The values of false-positive report probability were <0.015 for six SNPs, suggesting that the reported associations are less probably to be false. This study provides robust evidence for associations between genetic variants of CYP2E1 and NPC risk. PMID- 19805576 TI - Role of the nucleoplasmin 2 C-terminal domain in the formation of nucleolus-like bodies in mouse oocytes. AB - Nucleolus-like bodies (NLBs) are characteristic structures found in the germinal vesicles of mammalian oocytes. Although these structures are essential for embryonic development, their composition, precise function, and mechanism of formation have not been elucidated. Here, we used immunoblotting and EGFP fusion protein fluorescence to demonstrate that murine nucleoplasmin 2 (NPM2) is a component of mouse NLBs and that the targeting of NPM2 to NLBs is regulated by a lysine-rich, 16-aa C-terminal motif (K-rich motif). When the K-rich motif was fused to another nuclear protein, MafG, the resultant fusion protein accumulated in NLBs but not in the nucleoli of somatic cells, suggesting that the K-rich motif functions to target NPM2 specifically to NLBs. To investigate the role of the K-rich motif in NLB formation, we replaced the endogenous NPM2 in growing oocytes with a mutant NPM2 protein lacking the K-rich motif (NPM2(C16del)). Growing oocytes surrounded by granulosa layers were coinjected with NPM2(C16del) mRNA and with small-interfering RNA targeting NPM2 (siNpm2), which was used to degrade the endogenous NPM2 mRNA. After culture in vitro, the NLBs in the resulting full-grown oocytes were significantly smaller than those in control oocytes that had been coinjected with siNpm2 and NPM2 mRNA, indicating that the K rich motif is necessary for NLB development. Together, these results suggest that NPM2 targeting of NLBs is regulated by the K-rich motif and is essential for the formation of NLBs. PMID- 19805577 TI - ATP modulates Ca2+ uptake by TRPV6 and is counteracted by isoform-specific phosphorylation. AB - Ca(2+) homeostasis requires balanced uptake and extrusion, and dysregulation leads to disease. TRPV6 channels are homeostasis regulators, are upregulated in certain cancers, and show an unusual allele-specific evolution in humans. To understand how Ca(2+) uptake can be adapted to changes in metabolic status, we investigate regulation of Ca(2+)-influx by ATP and phosphorylation. We show that ATP binds to TRPV6, reduces whole-cell current increments, and prevents channel rundown with an EC(50) of 380 microM. By using both biochemical binding studies and patch-clamp analyses of wild-type and mutant channels, we have mapped one relevant site for regulation by ATP to residues within the ankyrin repeat domain (ARD) and identify an additional C-terminal binding region. Stimulation of PKC largely prevented the effects of ATP. This regulation requires PKC(betaII) and defined phosphorylation sites within the ARD and the C-terminus. Both regulatory sites act synergistically to constitute a novel mechanism by which ATP stabilizes channel activity and acts as a metabolic switch for Ca(2+) influx. Decreases in ATP concentration or activation of PKC(betaII) disable regulation of the channels by ATP, rendering them more susceptible to inactivation and rundown and preventing Ca(2+) overload. PMID- 19805578 TI - Genome-wide mapping of myosin protein-RNA networks suggests the existence of specialized protein production sites. AB - Motor proteins can organize posttranscriptional processes by transporting ribonucleoprotein complexes to specific locations. To investigate a possible role of myosin proteins in gene expression control, I have identified mRNAs associated with five myosin heavy chains in the fission yeast Schizosaccharomyces pombe, by purifying the proteins and identifying bound transcripts using DNA microarrays. Each myosin coimmunoprecipitated with 5-13 different mRNAs (approximately 0.1 0.2% of all genes), including those encoding four different myosin heavy chains. Moreover, one of the myosins (Myo1) interacted with mRNAs encoding components of the cortical actin cytoskeleton. These interactions were not observed in control immunoprecipitates. A myosin-specific chaperone (Rng3) that interacts cotranslationally with myosin mRNAs was essential for the association between myosin proteins and transcripts but not between Myo1 and other mRNAs. Finally, proteins encoded by the Myo1-associated mRNAs immunoprecipitated each other's transcripts, but not myosin mRNAs. These interactions suggest the existence of two distinct myosin-containing ribonucleoprotein complexes: those containing myosin mRNAs and those associated with Myo1. They are distinguished by their mRNA composition, requirement for the Rng3 chaperone and the presence of nonmyosin cytoskeletal proteins. I propose that these complexes represent specialized sites for the production of myosin proteins and the assembly of cytoskeletal components, respectively. PMID- 19805579 TI - FKBP12.6-knockout mice display hyperinsulinemia and resistance to high-fat diet induced hyperglycemia. AB - FK506 binding protein 12.6 kDa (FKBP12.6), a protein that regulates ryanodine Ca(2+) release channels, may act as an important regulator of insulin secretion. In this study, the role of FKBP12.6 in the control of insulin secretion and blood glucose is clarified using FKBP12.6(-/-) mice. FKBP12.6(-/-) mice showed significant fed hyperinsulinemia but exhibited normoglycemia, fasting normoinsulinemia, and normal body weight compared with wild-type (WT) littermate control mice. Deletion of FKBP12.6 resulted in enhanced glucose-stimulated insulin secretion (GSIS) both in vivo and in vitro, a result that is due to enhanced glucose-induced islet Ca(2+) elevation. After a high-fat dietary challenge (HF diet) for 3 mo, FKBP12.6(-/-) mice displayed higher body weight, hyperinsulinemia, and lower fed blood glucose concentrations compared with WT mice. FKBP12.6(-/-) mice displayed hyperinsulinemia, and resistance to HF diet induced hyperglycemia, suggesting that FKBP12.6 plays an important role in insulin secretion and blood glucose control, and raising the possibility that it may be a potential therapeutic target for the treatment of type 2 diabetes. PMID- 19805580 TI - Glucose-6-phosphate dehydrogenase-deficient mice have increased renal oxidative stress and increased albuminuria. AB - Glucose-6-phosphate dehydrogenase (G6PD) is the rate-limiting enzyme of the pentose phosphate pathway and the principal source of NADPH, a major cellular reductant, and is central to cell survival. Our previous work showed that diabetes and increased aldosterone are acquired forms of G6PD deficiency, leading to decreased G6PD activity and NADPH levels and damage to kidney tissue and endothelial cells. In this study, G6PD-deficient mice were studied to test the hypothesis that decreased G6PD activity per se can cause changes similar to those seen in the acquired conditions of G6PD deficiency. Results show that as compared with control mice, G6PD-deficient mice had increased oxidative stress, as manifested by decreased NADPH levels and decreased GSH levels, and increased markers of lipid peroxidation. G6PD-deficient mice had increased protein kinase C activity, increased nuclear factor-kappaB activity, and increased urinary albumin levels, all of which is similar to changes seen in diabetic mice. Changes persisted as the mice aged, as old G6PD-deficient mice (17-20 mo) had higher urine albumin levels and also had evidence for increased apoptosis in the renal cortex. These results show that decreased G6PD activity per se is sufficient to cause changes similar to those seen in diabetic mice. PMID- 19805581 TI - Treatment of scaphoid waist nonunions with an avascular proximal pole and carpal collapse. Surgical technique. AB - BACKGROUND: Surgically, it is difficult to achieve union of a scaphoid nonunion that is associated with osteonecrosis of the proximal pole, and those with carpal collapse are especially difficult to treat. A variety of vascularized bone grafts can be used. The purpose of this study was to compare the effectiveness of two types of vascularized bone graft -- a distal radial pedicle graft and a free vascularized medial femoral condyle graft -- in the treatment of scaphoid waist nonunions associated with proximal pole osteonecrosis and carpal collapse. METHODS: A retrospective review was conducted at two institutions to identify all patients with a scaphoid waist nonunion associated with an avascular proximal pole and carpal collapse. Between January 1994 and June 2006, twenty-two such nonunions were identified in twenty-two patients. Ten were treated with a distal radial pedicle vascularized graft and twelve, with a free vascularized medial femoral condyle graft. Patient demographics were similar between the groups, and the duration of follow-up averaged twelve months. Union was determined with use of plain radiographs and computed tomography or trispiral tomograms. In addition, carpal angles, time to union, union rates, and complications were recorded. RESULTS: Four of the ten nonunions treated with the distal radial pedicle graft healed, at a median of nineteen weeks, and all twelve nonunions treated with the free medial femoral condyle graft healed, at a median of thirteen weeks. The rate of union was significantly higher (p = 0.005) and the median time to healing was significantly shorter (p < 0.001) for the nonunions treated with the medial femoral condyle graft. CONCLUSIONS: A vascularized interposition graft from the medial femoral condyle is the recommended vascularized bone graft for the surgical treatment of scaphoid waist nonunion with avascularity of the proximal pole and carpal collapse. PMID- 19805582 TI - Long-term follow-up of radial shortening osteotomy for Kienbock disease. Surgical technique. AB - BACKGROUND: Three previous studies have investigated the long-term outcome of radial osteotomy in the treatment of Kienbock disease. However, none used patient based assessment. The purpose of this study was to investigate the long-term clinical and radiographic outcomes of this osteotomy, including the subjective evaluation of the patient with use of the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire. METHODS: A DASH questionnaire was sent to nineteen patients with Kienbock disease who had undergone a radial shortening osteotomy, and thirteen replied. The mean age at the time of surgery was thirty-nine years. On the basis of the Lichtman classification, six patients had stage-II, four had stage-IIIA, and three had stage-IIIB disease. Prior to surgery, ulnar variance was positive in six patients, neutral in four, and negative in three. The mean duration of follow-up was twenty-one years. Clinical evaluation, including calculation of the modified Mayo wrist score, and radiographic evaluation were also performed on twelve of the thirteen patients. RESULTS: The mean DASH score was 8 points (range, 0 to 23 points), and patient satisfaction was high. Compared with the findings in the contralateral wrist, the mean range of motion was 81% in flexion and 82% in extension and mean grip strength was 88%. The mean modified Mayo wrist score was 83 points, and the clinical results were excellent in six patients, good in five, and moderate in one. The DASH scores tended to be worse in patients with Lichtman stage-IIIB disease. Follow-up radiographs revealed that the Lichtman stage had progressed in six of the twelve patients. CONCLUSIONS: Although most patients had mild wrist pain, patient satisfaction and the clinical results were satisfactory following a radial shortening osteotomy. This procedure is a reliable long-term treatment for Lichtman stage-II and IIIA disease and may be a reasonable option for patients with stage-IIIB disease. PMID- 19805583 TI - Traumatic valgus instability of the elbow: pathoanatomy and results of direct repair. Surgical technique. AB - BACKGROUND: The medial collateral ligament provides valgus stability to the elbow. The purpose of the present study was to describe the pathoanatomy of acute traumatic medial collateral ligament ruptures and to report the rationale and results of direct repair. METHODS: Between 1996 and 2006, eleven athletes presented with acute rupture of the medial collateral ligament of the elbow and no history of dislocation. Three patients had received steroid injections for the treatment of medial epicondylitis, but none had a history of medial elbow insufficiency. All patients demonstrated gross valgus instability on clinical examination and medial joint space widening on valgus stress radiographs. Complete avulsion of the medial collateral ligament from its humeral origin was documented with magnetic resonance imaging in all patients. Operative findings uniformly demonstrated avulsion of the flexor-pronator muscles with distal retraction. The underlying medial collateral ligament was avulsed in a sleeve like fashion from the denuded medial epicondyle. The ligament was directly reattached to its footprint. The avulsed flexor-pronator tendon was repaired to the residual tendon with use of interrupted figure-of-eight nonabsorbable sutures. All patients were followed for a minimum of sixteen months with serial clinical examinations, radiographs, and the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire. RESULTS: Full active range of motion was achieved in ten patients; the remaining patient had a 20 degrees flexion contracture. Three patients had acute ulnar nerve palsies at the time of the injury, and all three recovered complete motor and sensory function by six months after the injury. Nine of the eleven patients returned to competitive college athletics between four and six months. The mean DASH score at the time of the most recent follow-up was 6. CONCLUSIONS: Direct repair of an acute traumatic medial collateral ligament avulsion of the elbow reliably restores valgus stability, even in throwing athletes. PMID- 19805584 TI - Linked elbow replacement: a salvage procedure for distal humeral nonunion. Surgical technique. AB - BACKGROUND: Nonunion is a challenging and not uncommon complication of distal humeral fractures. Our long-term experience with linked semiconstrained total elbow arthroplasty as a salvage procedure for patients with distal humeral nonunion not amenable to internal fixation was investigated. METHODS: Ninety-one consecutive patients (ninety-two elbows) underwent total elbow arthroplasty for the treatment of a distal humeral nonunion, and the results were reviewed at a mean of 6.5 years postoperatively. Patients' charts and anteroposterior and lateral radiographs made prior to and immediately after the joint replacement and at the time of the latest follow-up were reviewed to identify intraoperative and postoperative complications, and radiographic evidence of loosening or bushing wear. The outcome measures consisted of prosthetic survival, with implant removal as the end point for failure, and the Mayo Elbow Performance Score (MEPS). RESULTS: At the time of the most recent follow-up, joint stability had been initially restored in all patients, including nine who had had a grossly flailed elbow. Sixty-seven (74%) of the patients had no pain or mild pain at the time of the latest follow-up, whereas seventy-nine patients (87%) had had moderate or severe pain prior to the surgery. While 85% (seventy-seven) of the ninety-one patients rated the outcome as better or much better, twenty patients (22%) had a fair or poor MEPS. A total of forty-four complications occurred in forty elbows, and there were thirty-two reoperations, twenty-three of which involved implant revision or removal. Factors that increased the risk of implant failure were a patient age of less than sixty-five years, two or more prior surgical procedures, and a history of infection. The rate of prosthetic survival without removal or revision for any reason was 96% at two years, 82% at five years, and 65% at both ten and fifteen years. CONCLUSIONS: Linked semiconstrained total elbow arthroplasty is a salvage procedure that can provide pain relief and restore motion and function in patients with a distal humeral nonunion that is not amenable to internal fixation. Substantial risk factors for failure include an age of less than sixty-five years, multiple previous surgical procedures, and any history of infection. PMID- 19805585 TI - The early effects of tendon transfers and open capsulorrhaphy on glenohumeral deformity in brachial plexus birth palsy. Surgical technique. AB - BACKGROUND: Persistent muscle imbalance and soft-tissue contractures can lead to progressive glenohumeral joint dysplasia in patients with brachial plexus birth palsy. The objective of the present investigation was to determine the effects of tendon transfers and open glenohumeral reduction on shoulder function and dysplasia in patients with preexisting joint deformity secondary to brachial plexus birth palsy. METHODS: Twenty-three patients with preexisting glenohumeral deformity underwent latissimus dorsi and teres major tendon transfers to the rotator cuff with concomitant musculotendinous lengthening of the pectoralis major and/or subscapularis and open glenohumeral joint reduction for the treatment of internal rotation contracture and external rotation weakness. Shoulder function was assessed with use of the modified Mallet classification system and the Active Movement Scale. Glenoid version and humeral head subluxation were quantified radiographically, and glenohumeral deformity was appropriately graded. The mean duration of clinical and radiographic follow-up was thirty-one and twenty-five months, respectively. RESULTS: Clinically, all patients demonstrated improved global shoulder function, with the mean aggregate Mallet score improving from 10 points preoperatively to 18 points postoperatively (p < 0.01). The mean modified Mallet score for external rotation improved from 2 to 4 (p < 0.01). Similarly, the mean Active Movement Scale score for external rotation improved from 3 to 6 (p < 0.01). The mean Mallet hand-to-spine score improved from 1 to 2 (p < 0.01). The mean Active Movement Scale score for internal rotation remained constant at 6. Radiographically, the mean glenoid version improved from -39 degrees preoperatively to -18 degrees postoperatively (p < 0.01). The mean percentage of the humeral head anterior to the middle of the glenoid similarly improved from 13% to 38% (p < 0.01). The mean glenohumeral deformity score improved from 3 to 2 (p < 0.01). Nineteen (83%) of the twenty three patients demonstrated glenohumeral remodeling; one patient had progressive worsening of glenohumeral deformity. CONCLUSIONS: Tendon transfers to the rotator cuff, combined with musculotendinous lengthenings and open reduction of the glenohumeral joint, improve global shoulder function and lead to glenohumeral joint remodeling in the majority of selected patients with mild-to-moderate preexisting glenohumeral dysplasia secondary to brachial plexus birth palsy. Future study of the long-term outcomes of these procedures will help to clarify the ultimate effect on glenohumeral joint function. PMID- 19805586 TI - Cervical disc arthroplasty compared with arthrodesis for the treatment of myelopathy. Surgical technique. AB - BACKGROUND: Although there have been case reports describing the use of cervical disc arthroplasty for the treatment of myelopathy, there is a concern that motion preservation may maintain microtrauma to the spinal cord, negatively affecting the clinical results. As we are not aware of any studies on the use of arthroplasty in this scenario, we performed a cross-sectional analysis of two large, prospective, randomized multicenter trials to evaluate the efficacy of cervical disc arthroplasty for the treatment of myelopathy. METHODS: The patients in the current study were a cohort of patients who were enrolled in the United States Food and Drug Administration Investigational Device Exemption studies of the Prestige ST and Bryan disc replacements (Medtronic, Memphis, Tennessee). The inclusion criteria were myelopathy and spondylosis or disc herniation at a single level from C3 to C7. Clinical outcome measures were collected preoperatively and at six weeks, three months, six months, twelve months, and twenty-four months postoperatively. RESULTS: A total of 199 patients were included in the present study; 106 patients (53%) underwent arthroplasty, whereas ninety-three (47%) underwent arthrodesis. The Neck Disability Index, Short Form-36 scores, and specific arm and neck pain scores improved significantly from baseline at all time points. Patients in all four groups had improvement in the postoperative neurological status and gait function; at twenty-four months after surgery, 90% (95% confidence interval, 77.8% to 96.6%) of the patients in the arthroplasty group and 81% (95% confidence interval, 64.9% to 92.0%) of those in the arthrodesis group had improvement in or maintenance of the neurological status in the Prestige ST trial and 90% (95% confidence interval, 75.8% to 97.1%) of the patients in the arthroplasty group and 77% (95% confidence interval, 57.7% to 90.1%) of those in the arthrodesis group had improvement in or maintenance of the neurological status in the Bryan trial. CONCLUSIONS: We found that patients in both the arthroplasty and arthrodesis groups had improvement following surgery; furthermore, improvement was similar between the groups, with no worsening of myelopathy in the arthroplasty group. While the findings at two years postoperatively suggest that arthroplasty is equivalent to arthrodesis for the treatment of cervical myelopathy for a single-level abnormality localized to the disc space, the present study did not evaluate the treatment of retrovertebral compression as occurs in association with ossification of the posterior longitudinal ligament, and we cannot comment on the treatment of this condition. PMID- 19805587 TI - Surgical treatment of main thoracic scoliosis with thoracoscopic anterior instrumentation. Surgical technique. AB - BACKGROUND: The surgical outcomes in patients with scoliosis at two years following anterior thoracoscopic spinal instrumentation and fusion have been reported. The purpose of this study was to evaluate the results at five years. METHODS: A consecutive series of forty-one patients with major thoracic scoliosis treated with anterior thoracoscopic spinal instrumentation was evaluated at regular intervals. Prospectively collected data included patient demographics, radiographic measurements, clinical deformity measures, pulmonary function, an assessment of intervertebral fusion, and the scores on the Scoliosis Research Society (SRS-24) outcomes instrument. Perioperative and postoperative complications were recorded. Patient data for the preoperative, two-year, and five-year postoperative time points were compared. In addition, a univariate analysis compared selected two-year radiographic, pulmonary function, and SRS-24 data of the study cohort and those of the patients lost to follow-up. RESULTS: Twenty-five (61%) of the original forty-one patients had five-year follow-up data and were included in the analysis. Between the two-year and five-year follow-up visits, no significant changes were observed with regard to the average percent correction of the major Cobb angle (56% +/- 11% and 52% +/- 14%, respectively), average total lung capacity as a percent of the predicted value (95% +/- 14% and 91% +/- 10%), and the average total SRS-24 score (4.2 +/- 0.4 and 4.1 +/- 0.7). Radio-graphic evaluation of intervertebral fusion at five years revealed convincing evidence of a fusion with remodeling and trabeculae present at 151 (97%) of the 155 instrumented motion segments. No postoperative infections or clinically relevant neurovascular complications were observed. Rod failure occurred in three patients, and three patients required a surgical revision with posterior spinal instrumentation and fusion. CONCLUSIONS: Thoracoscopic anterior instrumentation for main thoracic idiopathic scoliosis results in five-year outcomes comparable with those reported previously for open anterior and posterior techniques. The radiographic findings, pulmonary function, and clinical measures remain stable between the two and five-year follow-up time points. Thoracoscopic instrumentation provides a viable alternative to treat spinal deformity; however, the risks of pseudarthrosis, hardware failure, and surgical revision should be considered along with the advantages of limited muscular dissection and improved scar appearance. PMID- 19805588 TI - Avulsion of the proximal hamstring origin. Surgical technique. AB - BACKGROUND: The torn hamstring is a common athletic injury. The purpose of the present study was to review the clinical presentation of this injury, the diagnostic imaging findings, the surgical technique of reattachment, and the likely clinical outcome of surgery for the treatment of avulsion of the proximal hamstring origin. METHODS: Seventy-two consecutive reconstructions in seventy-one patients with avulsion of the proximal hamstring origin were performed at a single center. The mean age at the time of the operation was 40.2 years. The mean duration of follow-up was twenty-four months, and all patients with a minimum duration of follow-up of six months were included. There were no exclusions. Patients were independently reviewed, and the mean postoperative isotonic hamstring strength was compared with that on the uninjured side. RESULTS: Waterskiing was the most frequent cause of injury (twenty-one cases). The mean time between the injury and the operation was twelve months. The most common pathological finding was a complete avulsion of the proximal hamstring origin (sixty-three cases; 87.5%), with a mean retraction of 7 cm (range, 0 to 20 cm). The mean postoperative isotonic hamstring strength measured 84% (range, 43% to 122%) and the mean postoperative hamstring endurance measured 89% (range, 26% to 161%) when compared with the values on the contralateral side. CONCLUSIONS: It is important to distinguish proximal hamstring origin avulsions (for which we recommend early surgical repair) from the majority of hamstring muscle injuries (which respond well to nonoperative treatment). The present study suggests that, in cases of complete avulsion with hamstring retraction, a delay in surgical repair renders the repair more technically challenging, may increase the likelihood of sciatic nerve involvement, increases the need for postoperative bracing, and reduces postoperative outcome in terms of hamstring strength and endurance. Once the nature of the injury has been established, the surgical treatment of hamstring origin avulsions has predictable and satisfactory results. PMID- 19805589 TI - Biomechanical consequences of a tear of the posterior root of the medial meniscus. Surgical technique. AB - BACKGROUND: Tears of the posterior root of the medial meniscus are becoming increasingly recognized. They can cause rapidly progressive arthritis, yet their biomechanical effects are not understood. The goal of this study was to determine the effects of posterior root tears of the medial meniscus and their repairs on tibiofemoral joint contact pressure and kinematics. METHODS: Nine fresh-frozen cadaver knees were used. An axial load of 1000 N was applied with a custom testing jig at each of four knee-flexion angles: 0 degrees , 30 degrees , 60 degrees , and 90 degrees . The knees were otherwise unconstrained. Four conditions were tested: (1) intact, (2) a posterior root tear of the medial meniscus, (3) a repaired posterior root tear, and (4) a total medial meniscectomy. Fuji pressure-sensitive film was used to record the contact pressure and area for each testing condition. Kinematic data were obtained by using a robotic arm to record the position of the knees for each loading condition. Three-dimensional knee kinematics were analyzed with custom programs with use of previously described transformations. The measured variables were axial rotation, varus angulation, lateral translation, and anterior translation. RESULTS: In the medial compartment, a posterior root tear of the medial meniscus caused a 25% increase in peak contact pressure compared with that found in the intact condition (p < 0.001). Repair restored the peak contact pressure to normal. No difference was detected between the peak contact pressure after the total medial meniscectomy and that associated with the root tear. The peak contact pressure in the lateral compartment after the total medial meniscectomy was up to 13% greater than that for all other conditions (p = 0.026). Significant increases in external rotation and lateral tibial translation, compared with the values in the intact knee, were observed in association with the posterior root tear (2.98 degrees and 0.84 mm, respectively) and the meniscectomy (4.45 degrees and 0.80 mm, respectively), and these increases were corrected by the repair. CONCLUSIONS: This study demonstrated significant changes in contact pressure and knee joint kinematics due to a posterior root tear of the medial meniscus. Root repair was successful in restoring joint biomechanics to within normal conditions. PMID- 19805590 TI - Distal femoral extension osteotomy and patellar tendon advancement to treat persistent crouch gait in cerebral palsy. Surgical technique. AB - BACKGROUND: Hallmarks of a persistent crouched walking pattern exhibited by individuals with cerebral palsy usually include loss of an adequate plantar flexion/knee extension couple, hamstring and/or psoas tightness, or contracture in conjunction with quadriceps insufficiency. Traditional treatment addresses the muscle-tightness component, but not the contracture or the muscle insufficiency. This study was performed to evaluate the effectiveness of distal femoral extension osteotomy and/or patellar tendon advancement in the treatment of crouch gait in patients with cerebral palsy. METHODS: A retrospective, nonrandomized, repeated-measures design was used. Individuals with a diagnosis of cerebral palsy were included if they had had (1) a distal femoral extension osteotomy in combination with a distal patellar tendon advancement (thirty-three patients), (2) a distal femoral extension osteotomy without patellar tendon advancement (sixteen), or (3) a distal patellar tendon advancement only (twenty-four). All subjects were evaluated with preoperative and postoperative gait analysis. Gait, radiographic, strength, and functional measures were included in the analysis to assess changes in knee function. RESULTS: Seventy-three individuals met the criteria for inclusion. A single side was chosen for the analysis of each subject. Ninety percent of the subjects had additional, concurrent surgery. Improvements were noted in the index assessing the level of gait pathology and in functional variables across all groups, and pain was consistently decreased. All preoperative stress fractures healed. Strength levels were maintained across all groups. The Koshino index of patellar height improved from 1.4 to -2.3 in the group treated with patellar tendon advancement only and from 1.5 to -2.9 in the group treated with both osteotomy and tendon advancement. The range of knee flexion improved an average of 15 degrees to 20 degrees , and stance-phase knee flexion was restored to the typical range (9 degrees to 10 degrees ) in the groups that had advancement of the patellar tendon as part of the procedure. Individuals who underwent a distal femoral osteotomy only were still in a crouch (a mean of 31 degrees of knee flexion in midstance) at the final assessment. CONCLUSIONS: Inclusion of patellar tendon advancement is necessary to achieve optimal results in the surgical management of a persistent crouch gait exhibited by adolescents and young adults with cerebral palsy. When this procedure is done alone or in combination with a distal femoral extension osteotomy (for the treatment of a knee flexion contracture), knee function in gait can be restored to values within typical limits, with gains in community function. PMID- 19805591 TI - Hindfoot endoscopy for posterior ankle impingement. Surgical technique. AB - BACKGROUND: The surgical treatment of posterior ankle impingement is associated with a high rate of complications and a substantial time to recover. An endoscopic approach to the posterior ankle (hindfoot endoscopy) may lack these disadvantages. We hypothesized that hindfoot endoscopy causes less morbidity and facilitates a quick recovery compared with open surgery. METHODS: Fifty-five consecutive patients with posterior ankle impingement were treated with an endoscopic removal of bone fragments and/or scar tissue. The symptoms were caused by trauma (65%) or overuse (35%). All patients were enrolled in a prospective protocol. At baseline, the age, sex, work and sports activities, American Orthopaedic Foot and Ankle Society (AOFAS) hindfoot scores, and preinjury Tegner scores were determined for all patients. At the time of follow-up, AOFAS hindfoot scores and Tegner scores were assessed and the time to return to work and sports activities was determined. Complications were recorded. Patients scored the overall result as poor, fair, good, or excellent by means of a 4-point Likert scale. RESULTS: The median duration of follow-up was thirty-six months, and no patient was lost to follow-up. The median AOFAS hindfoot score increased from 75 points preoperatively to 90 points at the time of final follow-up. The median time to return to work and sports activities was two and eight weeks, respectively. At the time of follow-up, patients in the overuse group were more satisfied than those in the posttraumatic group, and the AOFAS hindfoot scores were higher in patients in the overuse group (median, 100 points) compared with patients in the posttraumatic group (median, 90 points). A complication occurred in one patient who had a temporary loss of sensation of the posteromedial aspect of the heel. CONCLUSIONS: The outcome after endoscopic treatment of posterior ankle impingement compares favorably with the results of open surgery reported in the literature. Hindfoot endoscopy appears to cause less morbidity than open ankle surgery and facilitates a quick recovery. Patients treated for posterior ankle impingement caused by overuse have better results than those treated following trauma. PMID- 19805592 TI - A comparison of two nonoperative methods of idiopathic clubfoot correction: the Ponseti method and the French functional (physiotherapy) method. Surgical technique. AB - BACKGROUND: In the treatment of idiopathic clubfeet, the Ponseti method and the French functional method have been successful in reducing the need for surgery. The purpose of this prospective study was to compare the results of these two methods at one institution. METHODS: Patients under three months of age with previously untreated idiopathic clubfeet were enrolled. All feet were rated for severity prior to treatment. After both techniques had been described to them, the parents selected the treatment method. Outcomes at a minimum of two years were classified as good (a plantigrade foot with, or without, a heel-cord tenotomy), fair (a plantigrade foot that had or needed to have limited posterior release or tibialis anterior transfer), or poor (a need for a complete posteromedial surgical release). Two hundred and sixty-seven feet in 176 patients treated with the Ponseti method and 119 feet in eighty patients treated with the French functional method met the inclusion criteria. RESULTS: The patients were followed for an average of 4.3 years. Both groups had similar severity scores before treatment. The initial correction rates were 94.4% for the Ponseti method and 95% for the French functional method. Relapses occurred in 37% of the feet that had initially been successfully treated with the Ponseti method. One-third of the relapsed feet were salvaged with further nonoperative treatment, but the remainder required operative intervention. Relapses occurred in 29% of the feet that had been successfully treated with the French functional method, and all required operative intervention. At the time of the latest follow-up, the outcomes for the feet treated with the Ponseti method were good for 72%, fair for 12%, and poor for 16%. The outcomes for the feet treated with the French functional method were good for 67%, fair for 17%, and poor for 16%. CONCLUSIONS: Nonoperative correction of an idiopathic clubfoot deformity can be maintained over time in most patients. Although there was a trend showing improved results with use of the Ponseti method, the difference was not significant. In our experience, parents select the Ponseti method twice as often as they select the French functional method. PMID- 19805594 TI - Cell phones and choice architecture. PMID- 19805596 TI - Disproportionate sales of crime guns among licensed handgun retailers in the United States: a case-control study. AB - OBJECTIVE: To determine risk factors among licensed firearm retailers for disproportionate sales of handguns that are later subjected to ownership tracing, generally after use in crime. DESIGN: Case-control; the study period was 1998 2003. Cases were all eligible firearm retailers whose handguns were later traced at a rate that significantly (p<0.05) exceeded the expected value. Controls were a 4:1 random sample of the remainder. Data were obtained from sales and tracing records for 1998-2003 and site visits conducted August-December, 2004. SUBJECTS AND SETTING: 60 cases and 240 controls, from the 573 retailers in California selling >or= 50 handguns annually during the study period. MAIN OUTCOME MEASURE: Status as a case. Odds ratios were used to measure relative risk. RESULTS: In multivariate analyses, cases had larger sales volumes, sold inexpensive handguns more often, had a higher percentage of sales denied because the prospective purchasers were prohibited from owning firearms, and were more likely to be in an urban area, in or near a city with a policy of tracing all recovered crime guns. The effects of several risk factors, including status as a pawnbroker and sales to law enforcement personnel, appeared to be mediated by purchaser characteristics for which denied sales are a proxy measure. CONCLUSIONS: A number of factors-most of them characteristics of the retailers or of their handgun purchasers, and most of them available in existing data-were linked to disproportionate sales of handguns that are later used in crime. PMID- 19805597 TI - Validation of a HOME Injury Survey. AB - BACKGROUND: There are no validated observational surveys to assess injury hazards in the home environment. OBJECTIVE: To evaluate the reproducibility and reliability of a survey quantifying home injury hazards for children. METHODS: A nested cohort of children in the intervention arm of the Home Observations and Measures of the Environment (HOME) Study trial were analysed. The number and density of hazards were quantified by research assistants in the homes of participants at a baseline visit (BHV) for four high-risk rooms (kitchen, main activity room, child's bathroom and child's bedroom) and stairways and later at an intervention planning visit (IPV) for the four high-risk rooms and entire household. Statistical analysis included Pearson correlation, Bland-Altman analysis of agreement, analysis of variance and kappa statistics. RESULTS: There were 163 households with measurements at BHV and IPV. The number and density of hazards for the four high-risk rooms correlated significantly between BHV and IPV (r = 0.50 and 0.75, respectively). The number and density of hazards for the four high-risk rooms correlated significantly with that for the whole household at the IPV (r = 0.17 and 0.52, respectively). The number of injury hazards was significantly higher in the kitchen than in the other high-risk rooms, whereas density was highest in the child's bathroom. Inter-rater reliability between research assistants, as measured by the kappa statistic, was excellent with a mean of 0.81. CONCLUSIONS: The HOME Injury Survey was a reliable and replicable tool for quantifying residential injury hazards. The density of injury hazards was a more stable and valid measure than the number of injury hazards. PMID- 19805598 TI - Histories including number of falls may improve risk prediction for certain non vertebral fractures in older men. AB - OBJECTIVE: To determine whether information on number of falls on a falls history screen predicts risk of non-vertebral and hip fracture. METHODS: A cohort of 5995 community-dwelling men aged 65 years and older (mean 73.7) was followed over 7.2 years for incident non-vertebral fractures. Cox proportional hazard models were used to calculate hazard ratios (HRs) (95% CI) for incident fracture comparing a history of one and two or more falls with no falls. Models were adjusted for age, clinic, body mass index, height, femoral neck bone mineral density and whether the participant had a non-trauma fracture after the age of 50. por=0.08 mg/dl was detected among 43% of non-firearm-related (61% of AI) and 33% of firearm-related (50% of AI) homicide victims. Non-firearm-related homicide rates were highest among AI men aged 25-34 years (31/100,000). Non-firearm-related homicide victims were more likely than firearm-related victims to be AI (adjusted odds ratio (AOR) 4.20; 95% CI 2.16 to 8.16) and female (AOR 2.05; 95% CI 1.27 to 3.31), and to have had a BAC >or=0.08 mg/dl (AOR 1.65; 95% CI 1.08 to 2.52). CONCLUSIONS: Homicide-prevention efforts among AIs in NM should focus on non-firearm-related homicides. The association between excessive drinking and non-firearm-related homicide should be further characterised. Continued surveillance for non-firearm related homicides will assist these efforts. PMID- 19805601 TI - Characteristics of adult male and female firearm suicide decedents: findings from the National Violent Death Reporting System. AB - OBJECTIVE: To examine the risk factors and precipitating circumstances associated with firearm suicide. METHODS: Data from the restricted National Violent Death Reporting System (2003-6) for 25 491 male and female suicide decedents aged 18 and older were analysed by multiple logistic regression to estimate the relative odds of firearm use with 95% CIs. RESULTS: Firearms were often used in male (58.1%) and female (31.2%) suicides. Among male decedents, older age, veteran status, residing in areas with higher rates of firearm availability, raised blood alcohol concentration, acute crisis and relationship problems were all associated with firearm use. Conversely, men with a diagnosis of a mental health problem, a history of suicide attempts or alcohol problems had lower odds of firearm use. Among female decedents, factors with a significant effect on firearm use included: being older, married, white and a veteran; residing in areas with higher rates of firearm availability; having an acute crisis; having experienced the death of a relative or friend; being depressed; and having relationship problems. Of note, women who had a treated DSM-IV-diagnosed problem, previous suicide attempts and physical health problems were less likely to use firearms. CONCLUSIONS: These findings challenge the conventional view that those who are severely depressed and suicidal are prone to highly lethal methods, such as firearms. Rather, firearms users may be reacting to acute situations. PMID- 19805602 TI - The effect of counting principal and secondary injuries on national estimates of motor vehicle-related trauma: a NEISS-AIP special study. AB - OBJECTIVE: To demonstrate the effect of including both principal and secondary injuries in the calculation of national estimates of non-fatal motor vehicle related injury, using the National Electronic Injury Surveillance System-All Injury Program (NEISS-AIP). METHODS: The setting was a stratified sample of 15 US hospital emergency departments selected among 50 NEISS-AIP hospitals which agreed to participate in the study. Non-fatal injury data from a special study of the 2004 NEISS-AIP were analysed which allowed up to five injuries to be coded per case. National estimates of number and rate of injuries for 2004 were calculated, first using principal injuries alone, then by including principal and secondary injuries. RESULTS: An estimated 4,833,626 principal and secondary injuries were sustained by the estimated 2,893,782 motor vehicle occupants involved in a crash and treated in US hospital emergency departments (EDs) in 2004. This represents a 67% increase in the total number of injuries compared with an estimate of principal injury alone. Incidence of contusions/abrasions and lower trunk injuries rose most steeply among broad injury types, and whiplash injury rose 18% in number and rate. A significantly lower percentage of cases with a single listed injury were hospitalised (5%) compared with those who sustained multiple injuries (8%). CONCLUSIONS: Based on an analysis of NEISS-AIP special study data, the inclusion of both principal and secondary injuries in national estimates of motor vehicle-related occupant injury would provide a more comprehensive report of non-fatal injuries treated in US hospital EDs. Other countries with ED-based surveillance systems could consider reporting multiple injuries when assessing injury count associated with motor vehicle trauma requiring ED care. PMID- 19805603 TI - Innovation in qualitative interviews: "Sharing Circles" in a First Nations community. AB - There is growing recognition that different research approaches are necessary to understand the complex interaction between individual and social processes that contribute to risk-taking and injuries. Therefore, qualitative studies have an important role in injury prevention research. This article describes qualitative research in general and outlines some of the ways qualitative research can add to our understanding of injury. It also describes the role, format and methods of interviews (person-to-person and focus groups) commonly performed in qualitative studies, and proposes a novel approach to interviewing that has special relevance and value in injury research with indigenous populations. This methodology adapts focus group methods to be consistent with the goals and procedures of the traditional First Nations communities' Sharing Circles. This adaptation provides a culturally appropriate and sensitive method of developing a deep and broad understanding of indigenous participants' verbal descriptions of their feelings, their experiences and their modes of reasoning. After detailing of this adaptation of the Sharing Circle as a vibrant and vital interview and analysis method, the use of Sharing Circle interview methodology will be illustrated in a study investigating how an Alberta First Nations community experiences and deals with disproportionate levels of injuries arising from impaired driving, outlining important findings uncovered using this novel interviewing method. These findings have been informative to First Nations communities themselves, have informed policy makers provincially and nationally, and have instigated culturally appropriate intervention techniques for Canadian First Nations communities. PMID- 19805604 TI - The role of alcohol in unintentional falls among young and middle-aged adults: a systematic review of epidemiological studies. AB - OBJECTIVE: To appraise the published epidemiological evidence quantifying the risk of falls associated with acute and usual alcohol consumption among young and middle-aged adults. DESIGN: Systematic review. METHODS: DATA SOURCES: searches of electronic databases (eg, Medline, EMBASE, CINAHL, PsycINFO, Scopus), websites of relevant organisations, major injury journals, reference lists of relevant articles, and contact with experts in the field. INCLUSION CRITERIA: epidemiological studies with an English language abstract investigating alcohol use as a risk factor (exposure) for unintentional falls or related injuries among individuals aged 25-60 years. Studies were critically appraised using the GATE LITE tool. Meta-analysis was not attempted because of the heterogeneity of the eligible studies. RESULTS: Four case-control, three cohort and one case-crossover study fulfilled the inclusion criteria. The studies showed an increased risk of unintentional falls among young and middle-aged adults with increasing exposure to alcohol use. However, the magnitude of this risk varied considerably across studies with most estimates being relatively imprecise. Modest evidence of a dose response relationship with acute alcohol use was observed. The association between usual alcohol use and fall risk was inconclusive, and evidence of a gender difference was inconsistent. CONCLUSIONS: Alcohol use appears to be an important risk factor for falls among young and middle-aged adults. Controlled studies with sufficient power that adjust effect estimates for potential confounders (eg, fatigue, recreational drug use) are required to determine the population-based burden of fall-related injuries attributable to alcohol. This can help inform and prioritize falls prevention strategies for this age group. PMID- 19805605 TI - Booster seat laws and child fatalities: a case-control study. AB - A case-control study examined, primarily, the association between booster seat laws and fatalities among children in frontal collisions and, secondarily, the association between booster seat laws and reported restraint use, and restraint use and child fatalities. Children who died in a crash in the US were cases, and children who survived a fatal crash were controls. Subjects were child passengers (4-8 years old) in the Fatality Analysis Reporting System Database, 1995-2005. In states with a booster seat law, children were less likely to die than in states without a law (OR 0.80; 95% CI 0.66 to 0.98). They were also more likely to be restrained (adjusted OR 1.59; 95% CI 1.21 to 2.09) and were more likely to be correctly restrained (adjusted OR 4.44; 95% CI 3.18 to 6.20). It is concluded that booster seat laws are associated with a decrease in child deaths and an increase in correct restraint use among children involved in a fatal crash in the USA. PMID- 19805606 TI - Prospective outcomes of injury study. AB - BACKGROUND: In New Zealand (NZ), 20% of adults report a disability, of which one third is caused by injury. No prospective epidemiological studies of predictors of disability following all-cause injury among New Zealanders have been undertaken. Internationally, studies have focused on a limited range of predictors or specific injuries. Although these studies provide useful insights, applicability to NZ is limited given the importance of NZ's unique macro-social factors, such as NZ's no-fault accident compensation and rehabilitation scheme, the Accident Compensation Corporation (ACC). OBJECTIVES: (1) To quantitatively determine the injury, rehabilitation, personal, social and economic factors leading to disability outcomes following injury in NZ. (2) To qualitatively explore experiences and perceptions of injury-related outcomes in face-to-face interviews with 15 Maori and 15 other New Zealanders, 6 and 12 months after injury. SETTING: Four geographical regions within NZ. DESIGN: Prospective cohort study with telephone interviews 1, 4 and 12 months after injury. PARTICIPANTS: 2500 people (including 460 Maori), aged 18-64 years, randomly selected from ACC's entitlement claims register (people likely to be off work for at least 1 week or equivalent). DATA: Telephone interviews, electronic hospital and ACC injury data. Exposures include demographic, social, economic, work-related, health status, participation and/or environmental factors. OUTCOME MEASURES: Primary: disability (including WHODAS II) and health-related quality of life (including EQ-5D). Secondary: participation (paid and unpaid activities), life satisfaction and costs. ANALYSIS: Separate regression models will be developed for each of the outcomes. Repeated measures outcomes will be modelled using general estimating equation models and generalised linear mixed models. PMID- 19805607 TI - Lowering the default speed limit in residential areas: opportunities for policy influence and the role of public health professionals. PMID- 19805608 TI - Prevention of falls and fall-related injuries in older people. PMID- 19805609 TI - From SAVIR. PMID- 19805610 TI - WHO releases Guidelines for trauma quality improvement programmes. PMID- 19805614 TI - Time-course study of the immunotoxic effects of the anticancer drug chlorambucil in the rat. AB - In 2005, the International conference on harmonization (ICH) recommended that all new human pharmaceuticals be tested for unintended immunomodulatory potential via a tiered approach. Included in this approach is a semiquantitative description of changes in the separate compartments of lymphoid tissue (also called enhanced histopathology). Chlorambucil was administered to Hanover Wistar rats at regular time points, followed by a treatment-free (recovery) period. Groups of treated and control animals were sacrificed regularly during both the treatment and recovery periods. Selected tissues were removed, weighed fresh and fixed in formalin, processed, and stained with hematoxylin and eosin. Blood samples and bone marrow smears were also obtained. With the use of enhanced histopathology, a description of the changes in lymphoid tissues and bone marrow was used as a means of assessing the susceptibility, and recovery, of the different lymphoid cell populations over time. A correlation with organ weights, flow cytometry data, and bone marrow cytology was achieved. The administration of chlorambucil in the Hanover Wistar rat provided a useful tool to examine the rate and sequence of changes in the lymphoid organs and bone marrow during treatment with, and the recovery from the effects of, a potent immunosuppressive agent. PMID- 19805615 TI - Platelet protein disulfide isomerase is localized in the dense tubular system and does not become surface expressed after activation. AB - Evidence is accumulating that circulating tissue factor (TF) contributes to the initiation of coagulation and the formation of fibrin. The majority of circulating TF is cryptic, and it has been suggested that close vicinity with anionic phospholipids on the cell surface increases the active conformation of TF. Two recent papers have shown that encryption of TF and initiation of coagulation are facilitated by the enzyme protein disulfide isomerase (PDI), possibly on the surface of activated platelets or endothelial cells. In this brief report, we demonstrate that the majority of PDI in platelets is intracellular where it is exclusively located in the dense tubular system. On activation, PDI remains confined to the intracellular stores of the dense tubular system and is neither released nor targeted to the cell surface. Similar results were obtained in endothelium where PDI remains exclusively localized in the endoplasmic reticulum, both at steady state and after thrombin stimulation. PMID- 19805616 TI - Functionally distinct subsets of human NK cells and monocyte/DC-like cells identified by coexpression of CD56, CD7, and CD4. AB - The lack of natural killer (NK) cell-specific markers, as well as the overlap among several common surface antigens and functional properties, has obscured the delineation between NK cells and dendritic cells. Here, novel subsets of peripheral blood CD3/14/19(neg) NK cells and monocyte/dendritic cell (DC)-like cells were identified on the basis of CD7 and CD4 expression. Coexpression of CD7 and CD56 differentiates NK cells from CD56+ monocyte/DC-like cells, which lack CD7. In contrast to CD7+CD56+ NK cells, CD7(neg)CD56+ cells lack expression of NK cell-associated markers, but share commonalities in their expression of various monocyte/DC-associated markers. Using CD7, we observed approximately 60% of CD4+CD56+ cells were CD7(neg) cells, indicating the actual frequency of activated CD4+ NK cells is much lower in the blood than previously recognized. Functionally, only CD7+ NK cells secrete gamma interferon (IFNgamma) and degranulate after interleukin-12 (IL-12) plus IL-18 or K562 target cell stimulation. Furthermore, using CD7 to separate CD56+ NK cells and CD56+ myeloid cells, we demonstrate that unlike resting CD7+CD56+ NK cells, the CD7(neg)CD56+ myeloid cells stimulate a potent allogeneic response. Our data indicate that CD7 and CD56 coexpression discriminates NK cells from CD7(neg)CD56+ monocyte/DC-like cells, thereby improving our ability to study the intricacies of NK-cell subset phenotypes and functions in vivo. PMID- 19805617 TI - Cell-cell cooperation at the T helper cell/mast cell immunological synapse. AB - It has been suggested that mast cells might serve, under certain circumstances, as antigen-presenting cells (APCs) for T cells. However, whether cognate interactions between mast cells and class II-restricted CD4(+) T cells actually occur is still an open question. We addressed this question by using peritoneal cell-derived mast cells (PCMCs) and freshly isolated peritoneal mast cells as APC models. Our results show that in vitro treatment of PCMCs with interferon-gamma and interleukin-4 induced surface expression of mature major histocompatibility complex class II molecules and CD86. When interferon-gamma/interleukin-4-primed PCMCs were used as APCs for CD4(+) T cells, they induced activation of effector T cells but not of their naive counterparts as evidenced by CD69 up-regulation, proliferation, and cytokine production. Confocal laser scanning microscopy showed that CD4(+) T cells formed immunological synapses and polarized their secretory machinery toward both antigen-loaded PCMCs and freshly isolated peritoneal mast cells. Finally, on cognate interaction with CD4(+) T cells, mast cells lowered their threshold of activation via FcepsilonRI. Our results show that mast cells can establish cognate interactions with class II-restricted helper T cells, implying that they can actually serve as resident APCs in inflamed tissues. PMID- 19805618 TI - {beta}2 Glycoprotein I ({beta}2GPI) binds platelet factor 4 (PF4): implications for the pathogenesis of antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS) is an autoimmune thrombophilia characterized by arterial/venous thrombosis and/or pregnancy morbidity in the presence of antiphospholipid antibodies that mainly recognize beta2 glycoprotein I (beta2GPI). To investigate potential platelet ligands of beta2GPI, platelet membrane proteins from healthy persons and patients with APS were passed through a beta2GPI-affinity column. By using mass spectrometry, platelet factor 4 (PF4) appeared as the dominant beta2GPI binding protein. PF4 could bind in vitro, with high-affinity, recombinant beta2GPI, and the binding was abrogated by soluble beta2GPI. Coprecipitation experiments further confirmed this interaction. In silico molecular docking showed that PF4 tetramers can bind 2 beta2GPI molecules simultaneously. Size exclusion chromatography confirmed that anti-beta2GPI antibodies selectively interact with complexes composed of (beta2GPI)(2) (PF4)(4). In addition, as shown by the beta2GPI antigenicity evaluation, the reactivity of APS sera was higher against PF4-beta2GPI complex than against beta2GPI alone. On complex formation, anti-beta2GPI-beta2GPI-PF4 significantly induced platelet p38MAPK phosphorylation and TXB2 production, mainly through F(ab')(2) fragments of antibodies. In summary, this study makes evident that beta2GPI forms stable complexes with PF4, leading to the stabilization of beta2GPI dimeric structure that facilitates the antibody recognition. This interaction can probably be involved in the procoagulant tendency of APS. PMID- 19805619 TI - Expression profiling of a hemopoietic cell survival transcriptome implicates osteopontin as a functional prognostic factor in AML. AB - Deregulated cell survival programs are a classic hallmark of cancer. We have previously identified a serine residue (Ser585) in the betac subunit of the granulocyte-macrophage colony-stimulating factor receptor that selectively and independently promotes cell survival. We now show that Ser585 phosphorylation is constitutive in 20 (87%) of 23 acute myeloid leukemia (AML) patient samples, indicating that this survival-only pathway is frequently deregulated in leukemia. We performed a global expression screen to identify gene targets of this survival pathway and report a 138-gene betac Ser585-regulated transcriptome. Pathway analysis defines a gene network enriched for PI3-kinase target genes and a cluster of genes involved in cancer and cell survival. We show that one such gene, osteopontin (OPN), is a functionally relevant target of the Ser585-survival pathway as shown by siRNA-mediated knockdown of OPN expression that induces cell death in both AML blasts and CD34(+)CD38(-)CD123(+) leukemic progenitors. Increased expression of OPN at diagnosis is associated with poor prognosis with multivariate analysis indicating that it is an independent predictor of overall patient survival in normal karyotype AML (n = 60; HR = 2.2; P = .01). These results delineate a novel cytokine-regulated Ser585/PI3-kinase signaling network that is deregulated in AML and identify OPN as a potential prognostic and therapeutic target. PMID- 19805620 TI - Depletion of the C3 component of complement enhances the ability of rituximab coated target cells to activate human NK cells and improves the efficacy of monoclonal antibody therapy in an in vivo model. AB - Growing evidence indicates antibody-dependent cellular cytotoxicity (ADCC) contributes to the clinical response to monoclonal antibody (mAb) therapy of lymphoma. Recent in vitro analysis suggests C3b can inhibit mAb-induced natural killer (NK)-cell activation and ADCC. Further studies were conducted to assess the effect of C3 depletion on mAb-induced NK activation and therapy of lymphoma. Normal human serum inhibited the ability of rituximab-coated lymphoma cells to activate NK cells as previously reported. Serum did not inhibit NK-cell activation when it was preincubated with cobra venom factor (CVF) to deplete C3. Similar results were found when transudative pleural fluid or nonmalignant ascites was used as surrogates for extravascular fluid, suggesting the inhibitory effect of complement may be present in the extravascular compartment, in which many malignant lymphocytes reside. In vivo, C3 was depleted before mAb treatment in a syngeneic murine model of lymphoma. Survival of lymphoma-bearing mice after treatment with CVF plus mAb and with a human C3 derivative with CVF-like functions (HC3-1496) plus mAb was both superior to that of mAb alone. These studies show that complement depletion enhances NK-cell activation induced by rituximab-coated target cells and improves the efficacy of mAb therapy in a murine lymphoma model. PMID- 19805621 TI - Attention training for school-aged children with ADHD: results of an open trial. AB - OBJECTIVE: The article discusses a feasibility study conducted to examine whether Pay Attention!, an intervention training sustained, selective, alternating, and divided attention, could be utilized in a clinical setting with children diagnosed with ADHD, and whether children who received the intervention made attention and executive functioning gains. METHOD: After a diagnostic and baseline evaluation, 23 school-aged children with ADHD participate in up to 16 sessions of Pay Attention! and the outcomes are evaluated. RESULTS: Results show the intervention is feasible to administer and acceptable to participants. Parents and clinicians rate fewer ADHD symptoms following the intervention and report improvements in executive function. Child performance on neuropsychological tests showed improvements in fluid reasoning and cognitive flexibility and working memory. CONCLUSION: The findings suggest that a randomized clinical trial of Pay Attention! is warranted to investigate its viability as a treatment for attention and executive functioning deficits in ADHD. PMID- 19805622 TI - Maternal anxiety and attention problems in children at 5 and 14 years. AB - OBJECTIVE: This study examines the association between maternal anxiety from pregnancy to 5 years and child attention problems at 5 and 14 years. METHOD: Birth cohort of 3,982 individuals born in Brisbane between 1981 and 1983 are assessed. Self-reported measures of maternal anxiety are assessed at four time points. Maternal reports of child attention problems using Achenbach's Child Behavior Checklist are assessed at 5 and 14 years. RESULTS: Children of mothers experiencing anxiety during or after pregnancy are at greater risk of experiencing attention problems at 5 and 14 years. After adjusting for maternal age and child's gender, antenatal anxiety is strongly associated with persistent attention problems (OR = 3.65, 95% CI = 2.19, 6.07). Children with chronically anxious mothers are 5.67 (95% CI = 3.56, 9.03) times more likely to have persistent attention problems. These associations remain consistent after adjusting for potential confounders. CONCLUSIONS: Maternal anxiety appears to increase the rate of child attention problems and identifies a need for treatment programs to have a dual focus-the mother and her child. PMID- 19805623 TI - Shared and nonshared symptoms in youth-onset psychosis and ADHD. AB - OBJECTIVE: We compared ratings of behavior and attention problems between youth onset psychosis and ADHD, two disorders in which attentional impairments play a key role, and examined the effect of psychostimulant use on age of onset in psychosis. METHOD: Parent and teacher ratings of behavioral problems and ADHD symptoms were collected using the Achenbach CBCL, TRF, and SNAP-IV Teacher Rating Scales on 42 participants with psychosis, 36 with ADHD and 57 controls (ages 8 19). RESULTS AND CONCLUSIONS: Results suggested that academic, externalizing, and attention problems reflect symptoms shared between the disorders, whereas internalizing, social and thought problems reflect factors that differ between disorders. Furthermore, participants with psychosis who had been prescribed psychostimulants had a younger age of onset of psychotic symptoms than those who had not. This difference could reflect dissimilarities in symptom severity symptom between subgroups or potentially harmful effects of psychostimulants in individuals predisposed to develop psychosis. PMID- 19805624 TI - Secretory phospholipase A2 type III enhances alpha-secretase-dependent amyloid precursor protein processing through alterations in membrane fluidity. AB - In the non-amyloidogenic pathway, amyloid precursor protein (APP) is cleaved by alpha-secretases to produce alpha-secretase-cleaved soluble APP (sAPP(alpha)) with neuroprotective and neurotrophic properties; therefore, enhancing the non amyloidogenic pathway has been suggested as a potential pharmacological approach for the treatment of Alzheimer's disease. Here, we demonstrate the effects of type III secretory phospholipase A(2) (sPLA(2)-III) on sAPP(alpha) secretion. Exposing differentiated neuronal cells (SH-SY5Y cells and primary rat neurons) to sPLA(2)-III for 24 h increased sAPP(alpha) secretion and decreased levels of Abeta(1-42) in SH-SY5Y cells, and these changes were accompanied by increased membrane fluidity. We further tested whether sPLA(2)-III-enhanced sAPP(alpha) release is due in part to the production of its hydrolyzed products, including arachidonic acid (AA), palmitic acid (PA), and lysophosphatidylcholine (LPC). Addition of AA but neither PA nor LPC mimicked sPLA(2)-III-induced increases in sAPP(alpha) secretion and membrane fluidity. Treatment with sPLA(2)-III and AA increased accumulation of APP at the cell surface but did not alter total expressions of APP, alpha-secretases, and beta-site APP cleaving enzyme. Taken together, these results support the hypothesis that sPLA(2)-III enhances sAPP(alpha) secretion through its action to increase membrane fluidity and recruitment of APP at the cell surface. PMID- 19805626 TI - Irina Conboy: making the old feel young again. Interview by Kendall Powell. PMID- 19805625 TI - Disruption of the C-terminal helix by single amino acid deletion is directly responsible for impaired cholesterol efflux ability of apolipoprotein A-I Nichinan. AB - Apolipoprotein A-I (apoA-I) Nichinan, a naturally occurring variant with DeltaE235 in the C terminus, is associated with low plasma HDL levels. Here, we investigated the tertiary structure, lipid-binding properties, and ability to induce cellular cholesterol efflux of apoA-I Nichinan and its C-terminal peptide. Thermal and chemical denaturation experiments demonstrated that the DeltaE235 mutation decreased the protein stability compared with wild type (WT). ApoA-I Nichinan exhibited capabilities to bind to or solubilize lipid vesicles that are intermediate to that of WT and a L230P/L233P/Y236P variant in which the C terminal alpha-helix folding is completely disrupted and forms relatively larger and unstable discoidal complexes, indicating that perturbation of the C-terminal alpha-helical structure by the DeltaE235 mutation leads to reduced lipid binding. Supporting this, apoA-I 209-241/DeltaE235 peptide showed significantly decreased ability to form alpha-helix both in the lipid-free and lipid-bound states, and reduced efficiency to solubilize vesicles. In addition, both apoA-I Nichinan and its C-terminal peptide exhibited reduced activity in ABCA1-mediated cellular cholesterol efflux. Thus, the disruption of the ability of the C-terminal region to form alpha-helix caused by the E235 deletion appears to be the important determinant of impaired lipid binding and cholesterol efflux ability and, consequently, the low plasma HDL levels of apoA-I Nichinan probands. PMID- 19805627 TI - Tim-Tipin dysfunction creates an indispensible reliance on the ATR-Chk1 pathway for continued DNA synthesis. AB - The Tim (Timeless)-Tipin complex has been proposed to maintain genome stability by facilitating ATR-mediated Chk1 activation. However, as a replisome component, Tim-Tipin has also been suggested to couple DNA unwinding to synthesis, an activity expected to suppress single-stranded DNA (ssDNA) accumulation and limit ATR-Chk1 pathway engagement. We now demonstrate that Tim-Tipin depletion is sufficient to increase ssDNA accumulation at replication forks and stimulate ATR activity during otherwise unperturbed DNA replication. Notably, suppression of the ATR-Chk1 pathway in Tim-Tipin-deficient cells completely abrogates nucleotide incorporation in S phase, indicating that the ATR-dependent response to Tim-Tipin depletion is indispensible for continued DNA synthesis. Replication failure in ATR/Tim-deficient cells is strongly associated with synergistic increases in H2AX phosphorylation and DNA double-strand breaks, suggesting that ATR pathway activation preserves fork stability in instances of Tim-Tipin dysfunction. Together, these experiments indicate that the Tim-Tipin complex stabilizes replication forks both by preventing the accumulation of ssDNA upstream of ATR Chk1 function and by facilitating phosphorylation of Chk1 by ATR. PMID- 19805628 TI - Relative contributions of chromatin and kinetochores to mitotic spindle assembly. AB - During mitosis and meiosis in animal cells, chromosomes actively participate in spindle assembly by generating a gradient of Ran guanosine triphosphate (RanGTP). A high concentration of RanGTP promotes microtubule nucleation and stabilization in the vicinity of chromatin. However, the relative contributions of chromosome arms and centromeres/kinetochores in this process are not known. In this study, we address this issue using cells undergoing mitosis with unreplicated genomes (MUG). During MUG, chromatin is rapidly separated from the forming spindle, and both centrosomal and noncentrosomal spindle assembly pathways are active. MUG chromatin is coated with RCC1 and establishes a RanGTP gradient. However, a robust spindle forms around kinetochores/centromeres outside of the gradient peak. When stable kinetochore microtubule attachment is prevented by Nuf2 depletion in both MUG and normal mitosis, chromatin attracts astral microtubules but cannot induce spindle assembly. These results support a model in which kinetochores play the dominant role in the chromosome-mediated pathway of mitotic spindle assembly. PMID- 19805629 TI - Glyburide inhibits the Cryopyrin/Nalp3 inflammasome. AB - Inflammasomes activate caspase-1 for processing and secretion of the cytokines interleukin-1beta (IL-1beta) and IL-18. Cryopyrin/NALP3/NLRP3 is an essential component of inflammasomes triggered by microbial ligands, danger-associated molecular patterns (DAMPs), and crystals. Inappropriate Cryopyrin activity has been incriminated in the pathogenesis of gouty arthritis, Alzheimer's, and silicosis. Therefore, inhibitors of the Nalp3 inflammasome offer considerable therapeutic promise. In this study, we show that the type 2 diabetes drug glyburide prevented activation of the Cryopyrin inflammasome. Glyburide's cyclohexylurea group, which binds to adenosine triphosphatase (ATP)-sensitive K(+) (K(ATP)) channels for insulin secretion, is dispensable for inflammasome inhibition. Macrophages lacking K(ATP) subunits or ATP-binding cassette transporters also activate the Cryopyrin inflammasome normally. Glyburide analogues inhibit ATP- but not hypothermia-induced IL-1beta secretion from human monocytes expressing familial cold-associated autoinflammatory syndrome associated Cryopyrin mutations, thus suggesting that inhibition occurs upstream of Cryopyrin. Concurrent with the role of Cryopyrin in endotoxemia, glyburide significantly delays lipopolysaccharide-induced lethality in mice. Therefore, glyburide is the first identified compound to prevent Cryopyrin activation and microbial ligand-, DAMP-, and crystal-induced IL-1beta secretion. PMID- 19805630 TI - Intraflagellar transport particle size scales inversely with flagellar length: revisiting the balance-point length control model. AB - The assembly and maintenance of eukaryotic flagella are regulated by intraflagellar transport (IFT), the bidirectional traffic of IFT particles (recently renamed IFT trains) within the flagellum. We previously proposed the balance-point length control model, which predicted that the frequency of train transport should decrease as a function of flagellar length, thus modulating the length-dependent flagellar assembly rate. However, this model was challenged by the differential interference contrast microscopy observation that IFT frequency is length independent. Using total internal reflection fluorescence microscopy to quantify protein traffic during the regeneration of Chlamydomonas reinhardtii flagella, we determined that anterograde IFT trains in short flagella are composed of more kinesin-associated protein and IFT27 proteins than trains in long flagella. This length-dependent remodeling of train size is consistent with the kinetics of flagellar regeneration and supports a revised balance-point model of flagellar length control in which the size of anterograde IFT trains tunes the rate of flagellar assembly. PMID- 19805631 TI - Molecular mechanisms that enhance synapse stability despite persistent disruption of the spectrin/ankyrin/microtubule cytoskeleton. AB - Loss of spectrin or ankyrin in the presynaptic motoneuron disrupts the synaptic microtubule cytoskeleton and leads to disassembly of the neuromuscular junction (NMJ). Here, we demonstrate that NMJ disassembly after loss of alpha-spectrin can be suppressed by expression of a Wld(S) transgene, providing evidence for a Wallerian-type degenerative mechanism. We then identify a second signaling system. Enhanced MAPK-JNK-Fos signaling suppresses NMJ disassembly despite loss of presynaptic alpha-spectrin or ankyrin2-L. This signaling system is activated after an acute cytoskeletal disruption, suggesting an endogenous role during neurological stress. This signaling system also includes delayed, negative feedback via the JNK phosphatase puckered, which inhibits JNK-Fos to allow NMJ disassembly in the presence of persistent cytoskeletal stress. Finally, the MAPK JNK pathway is not required for baseline NMJ stabilization during normal NMJ growth. We present a model in which signaling via JNK-Fos functions as a stress response system that is transiently activated after cytoskeletal disruption to enhance NMJ stability, and is then shut off allowing NMJ disassembly during persistent cytoskeletal disruption. PMID- 19805632 TI - Cdc42 antagonizes Rho1 activity at adherens junctions to limit epithelial cell apical tension. AB - In epithelia, cells are arranged in an orderly pattern with a defined orientation and shape. Cadherin containing apical adherens junctions (AJs) and the associated actomyosin cytoskeleton likely contribute to epithelial cell shape by providing apical tension. The Rho guanosine triphosphatases are well known regulators of cell junction formation, maintenance, and function. Specifically, Rho promotes actomyosin activity and cell contractility; however, what controls and localizes this Rho activity as epithelia remodel is unresolved. Using mosaic clonal analysis in the Drosophila melanogaster pupal eye, we find that Cdc42 is critical for limiting apical cell tension by antagonizing Rho activity at AJs. Cdc42 localizes Par6-atypical protein kinase C (aPKC) to AJs, where this complex limits Rho1 activity and thus actomyosin contractility, independent of its effects on Wiskott-Aldrich syndrome protein and p21-activated kinase. Thus, in addition to its role in the establishment and maintenance of apical-basal polarity in forming epithelia, the Cdc42-Par6-aPKC polarity complex is required to limit Rho activity at AJs and thus modulate apical tension so as to shape the final epithelium. PMID- 19805633 TI - Electron-tomographic analysis of intraflagellar transport particle trains in situ. AB - Intraflagellar transport (IFT) is the bidirectional movement of multipolypeptide particles between the ciliary membrane and the axonemal microtubules, and is required for the assembly, maintenance, and sensory function of cilia and flagella. In this paper, we present the first high-resolution ultrastructural analysis of trains of flagellar IFT particles, using transmission electron microscopy and electron-tomographic analysis of sections from flat-embedded Chlamydomonas reinhardtii cells. Using wild-type and mutant cells with defects in IFT, we identified two different types of IFT trains: long, narrow trains responsible for anterograde transport; and short, compact trains underlying retrograde IFT. Both types of trains have characteristic repeats and patterns that vary as one sections longitudinally through the trains of particles. The individual IFT particles are highly complex, bridged to each other and to the outer doublet microtubules, and are closely apposed to the inner surface of the flagellar membrane. PMID- 19805634 TI - A novel measurement index for antihypertensive medication burden and its use. PMID- 19805635 TI - Hypertension correlates with lenticulostriate arteries visualized by 7T magnetic resonance angiography. AB - Hypertension, a major risk factor for stroke, is associated with altered arterial anatomy and function; however, the limited resolution of current imaging techniques has restricted the in vivo study of microvascular changes in the brain. In this report, we quantitatively examined the lenticulostriate arteries in hypertensive patients using ultrahigh-field 7T MRI. We compared the number of stems and branches, curvature, and tortuosity of the lenticulostriate arteries by 3D time-of-flight magnetic resonance angiography among 20 hypertensive patients (mean age: 46.6+/-9.1 years) and 20 age-matched healthy subjects (mean age: 47.7+/-8.1 years). The average numbers of stems and branches in hypertensive patients were significantly less than those of healthy subjects (P<0.002). However, this difference was abolished in older volunteers (>45 years old), whereas the difference between young hypertensive patients (< or = 45 years old) and age-matched healthy controls was augmented by 55% for stems and 91% for branches (P=0.001). In comparison, there were no differences in the average curvature and tortuosity of the lenticulostriate arteries and no significant difference when corrected for smoking (P=0.064). In conclusion, our results showed that there was a substantial difference in the lenticulostriate arteries of hypertensive patients compared with healthy individuals when observed in vivo by ultrahigh-resolution 7T magnetic resonance angiography, and the difference was considerable in young subjects. PMID- 19805636 TI - Early progression of the autonomic dysfunction observed in pediatric type 1 diabetes mellitus. AB - To focus on early cardiac and vascular autonomic dysfunction that might complicate type 1 diabetes mellitus in children, we planned an observational, cross-sectional study in a population of 93 young patients, under insulin treatment, subdivided in 2 age subgroups (children: 11.5+/-0.4 years; adolescents: 19.3+/-0.2 years). Time and frequency domain analysis of RR interval and systolic arterial pressure variability provided quantitative indices of the sympatho-vagal balance regulating the heart period, of the gain of cardiac baroreflex, and of the sympathetic vasomotor control. Sixty-eight children of comparable age served as a reference group. At rest, systolic arterial pressure and the power of its low-frequency component were greater in patients than in controls, particularly in children (14.0+/-2.3 versus 3.1+/-0.3 mm Hg2). Moreover, baroreflex gain was significantly reduced in both subgroups of patients. Standing induced similar changes in the autonomic profiles of controls and patients. A repeat study after 1 year showed a progression in low-frequency oscillations of arterial pressure and a shift toward low frequency in RR variability. Data in young patients with type 1 diabetes mellitus show a significant increase in arterial pressure, a reduced gain of the baroreflex regulation of the heart period, and an increase of the low-frequency component of systolic arterial pressure variability, suggestive of simultaneous impairment of vagal cardiac control and increases of sympathetic vasomotor regulation. A repeat study after 1 year shows a further increase of sympathetic cardiac and vascular modulation, suggesting early progression of the autonomic dysfunction. PMID- 19805637 TI - Genetic silencing of Nox2 and Nox4 reveals differential roles of these NADPH oxidase homologues in the vasopressor and dipsogenic effects of brain angiotensin II. AB - The renin-angiotensin system exerts a tremendous influence over fluid balance and arterial pressure. Angiotensin II (Ang-II), the effector peptide of the renin angiotensin system, acts in the central nervous system to regulate neurohumoral outflow and thirst. Dysregulation of Ang-II signaling in the central nervous system is implicated in cardiovascular diseases; however, the mechanisms remain poorly understood. Recently we established that NADPH oxidase (Nox)-derived superoxide acting in the forebrain subfornical organ is critical in the physiological responses to central Ang-II. In addition, we have found that Nox2 and Nox4 are the most abundantly expressed Nox homologues within Ang-II-sensitive sites in the forebrain. To dissect out the functional importance and unique roles of these Nox enzymes in the pressor and dipsogenic effects of central Ang-II, we developed adenoviral vectors expressing small interfering RNA to selectively silence Nox2 or Nox4 expression in the subfornical organ. Our results demonstrate that both Nox2 and Nox4 are required for the full vasopressor effects of brain Ang-II but that only Nox2 is coupled to the Ang-II-induced water intake response. These studies establish the importance of both Nox2- and Nox4-containing NADPH oxidases in the actions of Ang-II in the central nervous system and are the first to reveal differential involvement of these Nox enzymes in the various physiological effects of central Ang-II. PMID- 19805638 TI - Angiotensin receptor blocker prevented beta-amyloid-induced cognitive impairment associated with recovery of neurovascular coupling. AB - Recent studies suggest that vascular risk factors play a considerable role in the development of Alzheimer disease. Furthermore, the use of antihypertensive drugs has been suggested to reduce the incidence of dementia, including Alzheimer disease. In this study, we examined the effects of an angiotensin receptor blocker, olmesartan, on beta-amyloid-induced cerebrovascular dysfunction and cognitive impairment. Oral administration of a low dose of olmesartan attenuated cerebrovascular dysfunction in young Alzheimer disease-model transgenic mice (APP23 mouse), without a reduction in the brain beta-amyloid level. Moreover, treatment of APP23 mice with olmesartan decreased oxidative stress in brain microvessels. Using an acute mouse model induced by ICV administration of beta amyloid 1-40, we assessed the effect of oral administration of olmesartan on spatial learning evaluated with the Morris water maze. Olmesartan significantly improved cognitive function independent of its blood pressure-lowering effect, whereas there was no improvement by other types of antihypertensive drugs (hydralazine and nifedipine). We found that pretreatment with a low dose of olmesartan completely prevented beta-amyloid-induced vascular dysregulation and partially attenuated the impairment of hippocampal synaptic plasticity. These findings suggest the possibility that amelioration of cerebrovascular dysfunction with an angiotensin receptor blocker could be a novel therapeutic strategy for the early stage of Alzheimer disease. PMID- 19805640 TI - Mediterranean-style diet effect on the structural properties of the erythrocyte cell membrane of hypertensive patients: the Prevencion con Dieta Mediterranea Study. AB - A currently ongoing randomized trial has revealed that the Mediterranean diet, rich in virgin olive oil or nuts, reduces systolic blood pressure in high-risk cardiovascular patients. Here, we present a structural substudy to assess the effect of a Mediterranean-style diet supplemented with nuts or virgin olive oil on erythrocyte membrane properties in 36 hypertensive participants after 1 year of intervention. Erythrocyte membrane lipid composition, structural properties of reconstituted erythrocyte membranes, and serum concentrations of inflammatory markers are reported. After the intervention, the membrane cholesterol content decreased, whereas that of phospholipids increased in all of the dietary groups; the diminishing cholesterol:phospholipid ratio could be associated with an increase in the membrane fluidity. Moreover, reconstituted membranes from the nuts and virgin olive oil groups showed a higher propensity to form a nonlamellar inverted hexagonal phase structure that was related to an increase in phosphatidylethanolamine lipid class. These data suggest that the Mediterranean style diet affects the lipid metabolism that is altered in hypertensive patients, influencing the structural membrane properties. The erythrocyte membrane modulation described provides insight in the structural bases underlying the beneficial effect of a Mediterranean-style diet in hypertensive subjects. PMID- 19805639 TI - Relationship of carotid distensibility and thoracic aorta calcification: multi ethnic study of atherosclerosis. AB - Stiffening of the central elastic arteries is one of the earliest detectable manifestations of adverse change within the vessel wall. Although an association between carotid artery stiffness and adverse events has been demonstrated, little is known about the relationship between stiffness and atherosclerosis. Even less is known about the impact of age, sex, and race on this association. To elucidate this question, we used baseline data from the Multi-Ethnic Study of Atherosclerosis (2000-2002). Carotid artery distensibility coefficient was calculated after visualization of the instantaneous waveform of the common carotid diameter using a high-resolution B-mode ultrasound. Thoracic aorta calcification was identified using noncontrast cardiac computed tomography. We found a strong association between decreasing distensibility coefficient (increasing carotid stiffness) and increasing thoracic aorta calcification, as well as a graded increase in the thoracic aorta calcification score (P<0.001). After controlling for age, sex, race, and traditional and emerging cardiovascular risk factors, individuals in the stiffest quartile had a prevalence ratio of 1.52 (95% CI: 1.15 to 2.00) for thoracic aorta calcification compared with the least stiff quartile. In exploratory analysis, carotid stiffness was more highly correlated with calcification of the aorta than calcification of the coronary arteries (rho=0.32 versus 0.22; P<0.001 for comparison). In conclusion, there is a strong independent association between carotid stiffness and thoracic aorta calcification. Carotid stiffness is more highly correlated with calcification of the aorta, a central elastic artery, than calcification of the coronary arteries. The prognostic significance of these findings requires longitudinal follow-up of the Multi-Ethnic Study of Atherosclerosis cohort. PMID- 19805641 TI - Hepatocyte gp130 deficiency reduces vascular remodeling after carotid artery ligation. AB - Inflammation and vascular remodeling are hallmarks of atherosclerosis, hypertension, and restenosis after angioplasty. Here we investigated the role of the hepatocyte gp130-dependent systemic acute phase response on vascular remodeling after carotid artery ligation. Mice with a hepatocyte-specific gp130 knockout on an apolipoprotein E(-/-) background (gp130-) were compared with control mice (gp130(flox)). Vascular remodeling was induced by permanent ligation of the left common carotid artery. This, in turn, activated the systemic acute phase reaction in gp130(flox) mice, as measured by serum amyloid A plasma levels, which was completely abrogated in gp130- mice (P<0.05). Morphometric analysis of the carotid artery revealed severe neointima formation and media thickening 28 days after ligation in gp130(flox) mice, which was suppressed in gp130- mice (P<0.01). Serial sections from gp130- carotid segments showed significantly less smooth muscle cell (SMC) proliferation and monocyte recruitment (P<0.01). To evaluate the impact of the gp130-dependent systemic acute phase response on SMCs, hepatocytes from gp130(flox) and gp130- mice were stimulated with interleukin 6. Interleukin 6-induced secretion of serum amyloid A was completely abolished in gp130- hepatocytes (P<0.01). Moreover, when stimulated with supernatants from gp130- hepatocytes, SMCs showed significantly less migration and proliferation compared with supernatants from gp130(flox) hepatocytes (P<0.01). Recombinant serum amyloid A induced SMC migration and proliferation (P<0.05) and serum amyloid A injection after carotid artery ligation restored vascular remodeling in gp130- mice (P<0.01). These results imply a critical role for the gp130-dependent systemic acute phase response for vascular inflammation and SMC migration, as well as proliferation, and, subsequently, for vascular remodeling. PMID- 19805642 TI - Environmental mercury exposure and blood pressure among Nunavik Inuit adults. AB - Epidemiological evidence suggests a negative impact of methylmercury on the cardiovascular system, but findings regarding the effect on blood pressure (BP) are not consistent. We aimed to study the impact of mercury levels on BP among Nunavik Inuit adults. The health survey Qanuippitaa? was conducted in Nunavik (northern Quebec, Canada), and data were obtained from 732 Inuit > or = 18 years of age. Anthropometric blood samples, as well as systolic BP and diastolic BP, were assessed. Pulse pressure (systolic BP-diastolic BP) was calculated. Mercury blood concentration was used as a biomarker of recent exposure. Simple relations between mercury and BP parameters were studied by using the Pearson correlation, whereas multiple regressions were performed to control for confounders. Mean age of the participants was 34.3 years (95% CI: 33.6 to 34.9 years). Systolic BP, diastolic BP, and pulse pressure means were 117 mm Hg (95% CI: 116 to 118 mm Hg), 73 mm Hg (95% CI: 72 to 74 mm Hg), and 43 mm Hg (95% CI: 42 to 44 mm Hg), respectively. Mercury mean was 50.2 nmol/L. In multivariable analyses, mercury was associated with systolic BP (beta=2.14; P=0.0004), whereas the association with diastolic BP was near the significance level (beta=0.96; P=0.069). In conclusion, mercury is associated with increasing BP and pulse pressure among Nunavik Inuit adults after considering the effect of fish nutrients (n-3 fatty acids and selenium) and other confounders. PMID- 19805643 TI - Time-dependent effects of low-dose aspirin on plasma renin activity, aldosterone, cortisol, and catecholamines. AB - Studies have shown that aspirin may decrease blood pressure when given at bedtime but not when administered on awakening. However, until now, a biologically plausible mechanism of this striking phenomenon was not revealed. We investigated the effect of 100 mg of aspirin administered at bedtime compared with administration on awakening on plasma renin activity and aldosterone levels over 24 hours and excretion of cortisol and catecholamines in 24-hour urine samples. A randomized, placebo-controlled, double-blind, crossover trial was performed in 16 grade 1 hypertensive subjects. During 2 periods of 2 weeks separated by a 4-week washout period, participants used aspirin both at morning and at night, which was blinded with placebo. After both periods, subjects were admitted for 24 hours to measure the aforementioned parameters. Aspirin intake at bedtime compared with on awakening reduced average (24-hour) plasma renin activity by 0.08 microg/L per hour (95% CI: 0.03 to 0.13 microg/L per hour; P=0.003) without affecting aldosterone levels (95% CI: -0.01 to 0.01 nmol/L; P=0.93). Cortisol excretion in 24-hour urine was 52 nmol/24 hours (95% CI: 5 to 99 nmol/24 hours; P=0.05) lower, and dopamine and norepinephrine excretions were 0.25 micromol/24 hours (95% CI: 0.01 to 0.48 micromol/24 hours; P=0.04) and 0.22 micromol/24 hours (95% CI: -0.03 to 0.46 micromol/24 hours; P=0.02) lower in patients treated with bedtime aspirin. In conclusion, aspirin taken at bedtime compared with on awakening significantly diminished 24-hour plasma renin activity and excretion of cortisol, dopamine, and norepinephrine in 24-hour urine. Decreased activity of these pressor systems forms a biologically plausible explanation for the finding that aspirin at night may reduce blood pressure, whereas aspirin at morning does not. PMID- 19805645 TI - Impact of nonadherence to antihypertensive therapy. PMID- 19805644 TI - Renal proximal tubular reabsorption is reduced in adult spontaneously hypertensive rats: roles of superoxide and Na+/H+ exchanger 3. AB - Proximal tubule reabsorption is regulated by systemic and intrinsic mechanisms, including locally produced autocoids. Superoxide, produced by NADPH oxidase enhances NaCl transport in the loop of Henle and the collecting duct, but its role in the proximal tubule is unclear. We measured proximal tubule fluid reabsorption (Jv) in WKY rats and compared that with Jv in the spontaneously hypertensive rat (SHR), a model of enhanced renal superoxide generation. Rats were treated with the NADPH oxidase inhibitor apocynin (Apo) or with small interfering RNA for p22(phox), which is the critical subunit of NADPH oxidase. Jv was lower in SHR compared with Wistar-Kyoto rats (WKY; WKY: 2.3+/-0.3 vs SHR: 1.1+/-0.2 nL/min per millimeter; n=9 to 11; P<0.001). Apo and small interfering RNA to p22(phox) normalized Jv in SHRs but had no effect in WKY rats. Jv was reduced in proximal tubules perfused with S-1611, a highly selective inhibitor of the Na(+)/H(+) exchanger 3, the major Na(+) uptake pathway in the proximal tubule, in WKY rats but not in SHRs. Pretreatment with Apo restored an effect of S-1611 to reduce Jv in the SHRs (SHR+Apo: 2.9+/-0.4 vs SHR+Apo+S-1611: 1.0+/-0.3 nL/min per millimeter; P<0.001). However, because expression of the Na(+)/H(+) exchanger 3 was similar between SHR and WKY rats, this suggests that superoxide affects Na(+)/H(+) exchanger 3 activity. Direct microperfusion of Tempol or Apo into the proximal tubule also restored Jv in SHRs. In conclusion, superoxide generated by NADPH oxidase inhibits proximal tubule fluid reabsorption in SHRs. This finding implies that proximal tubule fluid reabsorption is regulated by redox balance, which may have profound effects on ion and fluid homeostasis in the hypertensive kidney. PMID- 19805646 TI - Assessment of structural disease in the coronary microvasculature. PMID- 19805647 TI - Hereditary deficiency of gp91(phox) is associated with enhanced arterial dilatation: results of a multicenter study. AB - BACKGROUND: NADPH oxidase is believed to modulate arterial tone, but its role in humans is still unclear. The objective of this study was to evaluate whether NADPH oxidase is involved in flow-mediated arterial dilation (FMD). METHODS AND RESULTS: Twenty-five patients with hereditary deficiency of gp91(phox), the catalytic core of NADPH oxidase, (X-CGD), 25 healthy subjects, and 25 obese patients matched for sex and age were recruited. FMD, platelet gp91(phox), serum levels of nitrite and nitrate as markers of nitric oxide generation, oxidized low density lipoprotein, and urinary excretion of isoprostanes as markers of oxidative stress were determined. Platelet gp91(phox) expression was downregulated in X-CGD patients (1.0+/-0.8 mean fluorescence; P<0.001) and upregulated in obese patients (4.1+/-2.2 mean fluorescence; P=0.01) compared with healthy subjects (2.9+/-1.7 mean fluorescence). Urinary excretion of isoprostanes was reduced in X-CGD patients (41.7+/-33.3 pg/mg creatinine; P=0.04) and increased in obese patients (154.4+/-91 pg/mg creatinine; P<0.001) compared with healthy subjects (69.5+/-52.4 pg/mg creatinine). Obese patients had higher serum oxidized low-density lipoprotein than healthy subjects (35.3+/-6.7 versus 24.8+/ 9.8 U/L; P<0.001) and X-CGD patients (28.5+/-7.2 U/L; P<0.001). X-CGD patients had significantly higher FMD (14.7+/-5.9%) compared with healthy subjects (7.9+/ 2.5%; P<0.001); obese patients had lower FMD (5.3+/-3.0%; P=0.028) compared with healthy subjects. Serum nitrite and nitrate levels were significantly higher in patients with X-CGD (36.0+/-10.8 micromol/L; P=0.016) and lower in obese patients (9.3+/-11.0 micromol/L; P=0.001) compared with healthy subjects (27.1+/-19.1 micromol/L). Serum nitrite and nitrate levels significantly correlated with FMD (R(s)=0.403, P<0.001) and platelet gp91(phox) (R(s)=-0.515, P<0.001). FMD inversely correlated with platelet gp91(phox) (R(s)=-0.502, P<0.001) and isoprostanes (R(s)=-0.513, P<0.001). CONCLUSIONS: This study provides the first evidence that, in humans, gp91(phox) is implicated in the modulation of arterial tone. PMID- 19805648 TI - Activin A and follistatin-like 3 determine the susceptibility of heart to ischemic injury. AB - BACKGROUND: Transforming growth factor-beta family cytokines have diverse actions in the maintenance of cardiac homeostasis. Activin A is a member of this family whose regulation and function in heart are not well understood at a molecular level. Follistatin-like 3 (Fstl3) is an extracellular regulator of activin A protein, and its function in the heart is also unknown. METHODS AND RESULTS: We analyzed the expression of various transforming growth factor-beta superfamily cytokines and their binding partners in mouse heart. Activin betaA and Fstl3 were upregulated in models of myocardial injury. Overexpression of activin A with an adenoviral vector (Ad-actbetaA) or treatment with recombinant activin A protein protected cultured myocytes from hypoxia/reoxygenation-induced apoptosis. Systemic overexpression of activin A in mice by intravenous injection of Ad actbetaA protected hearts from ischemia/reperfusion injury. Activin A induced the expression of Bcl-2, and ablation of Bcl-2 by small interfering RNA abrogated its protective action in myocytes. The protective effect of activin A on cultured myocytes was abolished by treatment with Fstl3 or by a pharmacological activin receptor-like kinase inhibitor. Cardiac-specific Fstl3 knockout mice showed significantly smaller infarcts after ischemia/reperfusion injury that was accompanied by reduced apoptosis. CONCLUSIONS: Activin A and Fstl3 are induced in heart by myocardial stress. Activin A protects myocytes from death, and this activity is antagonized by Fstl3. Thus, the relative expression levels of these factors after injury is a determinant of cell survival in the heart. PMID- 19805649 TI - Absence of thrombospondin-2 causes age-related dilated cardiomyopathy. AB - BACKGROUND: The progressive shift from a young to an aged heart is characterized by alterations in the cardiac matrix. The present study investigated whether the matricellular protein thrombospondin-2 (TSP-2) may affect cardiac dimensions and function with physiological aging of the heart. METHODS AND RESULTS: TSP-2 knockout (KO) and wild-type mice were followed up to an age of 60 weeks. Survival rate, cardiac function, and morphology did not differ at a young age in TSP-2 KO compared with wild-type mice. However, >55% of the TSP-2 KO mice died between 24 and 60 weeks of age, whereas <10% of the wild-type mice died. In the absence of TSP-2, older mice displayed a severe dilated cardiomyopathy with impaired systolic function, increased cardiac dilatation, and fibrosis. Ultrastructural analysis revealed progressive myocyte stress and death, accompanied by an inflammatory response and replacement fibrosis, in aging TSP-2 KO animals, whereas capillary or coronary morphology or density was not affected. Importantly, adeno-associated virus-9 gene-mediated transfer of TSP-2 in 7-week old TSP-2 KO mice normalized their survival and prevented dilated cardiomyopathy. In TSP-2 KO animals, age-related cardiomyopathy was accompanied by increased matrix metalloproteinase-2 and decreased tissue transglutaminase-2 activity, together with impaired collagen cross-linking. At the cardiomyocyte level, TSP-2 deficiency in vivo and its knockdown in vitro decreased the activation of the Akt survival pathway in cardiomyocytes. CONCLUSIONS: TSP-2 expression in the heart protects against age-dependent dilated cardiomyopathy. PMID- 19805650 TI - Bleeding increases the risk of ischemic events in patients with peripheral arterial disease. AB - BACKGROUND: Patients with peripheral arterial disease are at high risk of ischemic events and therefore are treated with antithrombotics. In patients with coronary artery disease or cerebrovascular disease, bleeding is related to the subsequent occurrence of ischemic events. Our objective was to assess whether this is also the case in patients with peripheral arterial disease. METHODS AND RESULTS: All patients from the Dutch Bypass and Oral Anticoagulants or Aspirin (BOA) Study, a multicenter randomized trial comparing oral anticoagulants with aspirin after infrainguinal bypass surgery, were included. The primary outcome event was the composite of nonfatal myocardial infarction, nonfatal ischemic stroke, major amputation, and cardiovascular death. To identify major bleeding as an independent predictor for ischemic events, crude and adjusted hazard ratios with 95% confidence intervals were calculated with multivariable Cox regression models. From 1995 until 1998, 2650 patients were included with 101 nonfatal major bleedings. During a mean follow-up of 14 months, the primary outcome event occurred in 218 patients; 22 events were preceded by a major bleeding. The mean time between major bleeding and the primary outcome event was 4 months. Major bleeding was associated with a 3-fold increased risk of subsequent ischemic events (crude hazard ratio, 3.0; 95% confidence interval, 1.9 to 4.6; adjusted hazard ratio, 3.0; 95% confidence interval, 1.9 to 4.7). CONCLUSIONS: In patients with peripheral arterial disease, as in patients with coronary artery disease or cerebrovascular disease, major bleeding was independently associated with major ischemic complications. Without compromising the benefits of antithrombotics, these findings call for caution relative to the risks of major bleeding. PMID- 19805651 TI - Proteinuria, chronic kidney disease, and the effect of an angiotensin receptor blocker in addition to an angiotensin-converting enzyme inhibitor in patients with moderate to severe heart failure. AB - BACKGROUND: Chronic kidney disease (CKD) is an established risk factor for poor outcomes in heart failure (HF). Whether proteinuria provides additional prognostic information is not known. Renin-angiotensin blockade medications improve outcomes in HF but are underutilized in HF patients with renal dysfunction because of safety concerns and a lack of evidence of their effectiveness. METHODS AND RESULTS: In the Valsartan in Heart Failure Trial (Val HeFT), 5010 patients with class II, III, or IV heart failure were randomly assigned to receive valsartan or placebo. The 2 primary outcomes were death and first morbid event, defined as death, sudden death with resuscitation, hospitalization for HF, or administration of intravenous inotropic or vasodilator drugs for 4 hours or more without hospitalization. The study cohort was divided into subgroups according to the presence of CKD (estimated glomerular filtration rate <60 mL x min(-1) x 1.73 m(-2)) and proteinuria (positive dipstick). Multivariable Cox proportional hazards regression models were used to examine the relationships between study outcomes and proteinuria, including its interaction with CKD. The interaction between valsartan and CKD was also tested. The effect of valsartan on estimated glomerular filtration rate was estimated by generalized linear models, including tests of interactions between treatment and CKD. At baseline, CKD was found in 58% and dipstick-positive proteinuria in 8% of patients. Dipstick-positive proteinuria was independently associated with mortality (hazard ratio [HR] 1.28, 95% confidence interval [CI] 1.01 to 1.62, P=0.05) and first morbid event (HR 1.28, 95% CI 1.06 to 1.55, P=0.01). The increased risk of death associated with dipstick-positive proteinuria was similar for those with and without CKD (HR 1.26, 95% CI 0.96 to 1.66 versus HR 1.37, 95% CI 0.83 to 2.26; P=0.94), as was the hazard for first morbid event (HR 1.26, 95% CI 1.01 to 1.57 versus HR 1.42, 95% CI 0.98 to 2.07; P=0.71). Valsartan reduced estimated glomerular filtration rate compared with placebo to a similar extent (P=0.52) in the subgroups with CKD (mean reduction -3.6 mL x min(-1) x 1.73 m( 2)) and without CKD (mean reduction -4.0 mL x min(-1) x 1.73 m(-2)) and by -3.8 mL x min(-1) x 1.73 m(-2) in both groups combined. The beneficial effect of valsartan on first morbid events was similar in those with and without CKD (HR 0.86, 95% CI 0.74 to 0.99 versus HR 0.91, 95% CI 0.73 to 1.12; P=0.23) and was significant in the subgroup with CKD. The effect of valsartan on mortality did not differ in patients with and without CKD (HR 1.01, 95% CI 0.85 to 1.20 versus HR 0.91, 95% CI 0.69 to 1.25; P=0.08). CONCLUSIONS: CKD was common and dipstick positive proteinuria was infrequent in this sample of patients with HF. After controlling for other risk factors, including CKD, the relatively small subgroup with dipstick-positive proteinuria did have worse outcomes. Valsartan reduced the estimated glomerular filtration rate by the same amount in patients with and without CKD and reduced the risk of the first morbid event in patients with CKD, which suggests its beneficial effects in patients with HF and CKD. PMID- 19805652 TI - Assessment of microcirculatory remodeling with intracoronary flow velocity and pressure measurements: validation with endomyocardial sampling in cardiac allografts. AB - BACKGROUND: Intracoronary physiology techniques have been validated extensively for the assessment of epicardial stenoses but not for the lone study of coronary microcirculation. We performed a comparison between 4 intracoronary physiological indices with the actual structural microcirculatory changes documented in transplanted hearts. METHODS AND RESULTS: In 17 cardiac allograft patients without coronary stenoses, ECG, intracoronary Doppler flow velocity, and aortic pressure were digitally recorded before and during maximal hyperemia with a dedicated system. Postprocessing of data yielded 4 indices of microcirculatory status: coronary flow velocity reserve (2.13+/-0.59), instantaneous hyperemic diastolic velocity pressure slope (2.33+/-1.25 cm x s x (-1)mm Hg(-1)), coronary resistance index (1.65+/-0.88 mm Hg x cm(-1) x s(-1)), and coronary resistance reserve (2.36+/-0.65). Quantitative morphometry was performed in endomyocardial biopsies during the same hospital intake; arteriolar obliteration (76.57+/-6.95%) and density (2.00+/-1.22 arterioles per 1 mm(2)) and capillary density (645+/-179 capillaries per 1 mm(2)) were measured. Univariate regression analysis between intracoronary measurements and histological findings revealed that instantaneous hyperemic diastolic velocity-pressure slope correlated with arteriolar obliteration (r=0.58, P=0.014) and capillary density (r=0.60, P=0.012). Statistical adjustment revealed an independent contribution of arteriolar obliteration (beta=0.61, P=0.0009) and capillary density (beta=-0.60, P=0.0008) to instantaneous hyperemic diastolic velocity-pressure slope values, resulting in an excellent predictive model (r=0.84, P=0.0002). Coronary resistance index correlated only with capillary density (r=0.70, P=0.019). Relative indices (coronary flow velocity reserve and coronary resistance reserve) did not correlate significantly with arteriolar obliteration, capillary density, or arteriolar density. CONCLUSIONS: Intracoronary indices derived from pressure and flow, particularly instantaneous hyperemic diastolic velocity-pressure slope, appear to be superior to coronary flow velocity reserve in detecting structural microcirculatory changes. Both arteriolar obliteration and capillary rarefaction seem to influence microcirculatory hemodynamics independently. PMID- 19805653 TI - Adherence to antihypertensive medications and cardiovascular morbidity among newly diagnosed hypertensive patients. AB - BACKGROUND: Nonadherence to antihypertensive treatment is a common problem in cardiovascular prevention and may influence prognosis. We explored predictors of adherence to antihypertensive treatment and the association of adherence with acute cardiovascular events. METHODS AND RESULTS: Using data obtained from 400 Italian primary care physicians providing information to the Health Search/Thales Database, we selected 18,806 newly diagnosed hypertensive patients >or=35 years of age during the years 2000 to 2001. Subjects included were newly treated for hypertension and initially free of cardiovascular diseases. Patient adherence was subdivided a priori into 3 categories-high (proportion of days covered, >or=80%), intermediate (proportion of days covered, 40% to 79%), and low (proportion of days covered, or = 50,000/microL and/or age > or = 10 years. Randomly assigned therapies evaluated the impact of postinduction treatment intensification on outcome. We examined outcome and prognostic factors for 262 young adults with ALL. RESULTS: Five-year event-free and overall survival rates for young adult patients are 71.5% (SE, 3.6%) and 77.5% (SE, 3.3%), respectively. Rapid responder patients (< 25% bone marrow blasts on day 7) randomly assigned to augmented therapy had 5-year event-free survival of 81.8% (SE, 7%), as compared with 66.8% (SE, 6.7%) for patients receiving standard therapy (P = .07). One versus two interim maintenance and delayed intensification courses had no significant impact on event-free survival. WBC count more than 50,000/microL was an adverse prognostic factor. CONCLUSION: Young adult patients with ALL showing a rapid response to induction chemotherapy benefit from early intensive postinduction therapy but do not benefit from a second interim maintenance and delayed intensification phase. Given the excellent outcome with this chemotherapy, there seems to be no role for the routine use of allogeneic stem-cell transplantation in first remission for young adults with ALL. PMID- 19805690 TI - Risk of relapse of childhood acute lymphoblastic leukemia is predicted by flow cytometric measurement of residual disease on day 15 bone marrow. AB - PURPOSE: Speed of blast clearance is an indicator of outcome in childhood acute lymphoblastic leukemia (ALL). Availability of measurement of minimal residual disease (MRD) at an early time point with a reduced-cost method is of clinical relevance. In the AIEOP-BFM-ALL (Associazione Italiana Ematologia Oncologia Pediatrica and Berlin-Frankfurt-Munster Study Group) 2000 trial, patients were stratified by levels of polymerase chain reaction (PCR) MRD at day +33 and +78. AIEOP studied the prognostic impact of MRD measured by flow cytometry (FCM) at day 15 of induction therapy. PATIENTS AND METHODS: Bone marrow samples from 830 Italian patients were collected on day 15, after 14 days of steroids, and one dose of intrathecal methotrexate, vincristine, daunorubicine, and asparaginase. Cells were analyzed by four-color FCM for detection of leukemia-associated immunophenotypes. RESULTS: Three patient risk groups were identified by FCM: standard (< 0.1% blast cells; 42% of the total), intermediate (0.1 to < 10%; 47%), and high (> or = 10%; 11%). Their 5-year cumulative incidences of relapse were 7.5% (SE, 1.5), 17.5% (SE, 2.1), and 47.2% (SE, 5.9), respectively. In multivariate analysis, FCM was the most important prognostic factor among those available by day 15, with two-fold and five-fold increase in the risk of relapse compared with patients with less than 0.1%. PCR MRD, when added to the model, had significant prognostic impact; yet high levels of FCM MRD retained an independent ability to detect a significantly higher risk of relapse. CONCLUSION: Measurement of FCM MRD in day 15 bone marrow was the most powerful early predictor of relapse, applicable to virtually all patients; it may complement PCR MRD-based stratification including later time points, thus allowing additional treatment tailoring. PMID- 19805691 TI - Comparison of iodine-123 metaiodobenzylguanidine (MIBG) scan and [18F]fluorodeoxyglucose positron emission tomography to evaluate response after iodine-131 MIBG therapy for relapsed neuroblastoma. AB - PURPOSE: Children with relapsed neuroblastoma have poor survival. It is crucial to have a reliable method for evaluating functional response to new therapies. In this study, we compared two functional imaging modalities for neuroblastoma: metaiodobenzylguanidine (MIBG) scan for uptake by the norepinephrine transporter and [(18)F]fluorodeoxyglucose positron emission tomography (FDG-PET) uptake for glucose metabolic activity. PATIENTS AND METHODS: Patients enrolled onto a phase I study of sequential infusion of iodine-131 ((131)I) MIBG (NANT-2000-01) were eligible for inclusion if they had concomitant FDG-PET and MIBG scans. (131)I MIBG therapy was administered on days 0 and 14. For each patient, we compared all lesions identified on concomitant FDG-PET and MIBG scans and gave scans a semiquantitative score. RESULTS: The overall concordance of positive lesions on concomitant MIBG and FDG-PET scans was 39.6% when examining the 139 unique anatomic lesions. MIBG imaging was significantly more sensitive than FDG-PET overall and for the detection of bone lesions (P < .001). There was a trend for increased sensitivity of FDG-PET for detection of soft tissue lesions. Both modalities showed similar improvement in number of lesions identified from day 0 to day 56 scan and in semiquantitative scores that correlated with overall response. FDG-PET scans became completely negative more often than MIBG scans after treatment. CONCLUSION: MIBG scan is significantly more sensitive for individual lesion detection in relapsed neuroblastoma than FDG-PET, though FDG PET can sometimes play a complementary role, particularly in soft tissue lesions. Complete response by FDG-PET metabolic evaluation did not always correlate with complete response by MIBG uptake. PMID- 19805692 TI - Multinational, double-blind, phase III study of prednisone and either satraplatin or placebo in patients with castrate-refractory prostate cancer progressing after prior chemotherapy: the SPARC trial. AB - PURPOSE: This multinational, double-blind, randomized, placebo-controlled, phase III trial assessed the efficacy and tolerability of the oral platinum analog satraplatin in patients with metastatic castrate-refractory prostate cancer (CRPC) experiencing progression after one prior chemotherapy regimen. PATIENTS AND METHODS: Nine hundred fifty patients were randomly assigned (2:1) to receive oral satraplatin (n = 635) 80 mg/m(2) on days 1 to 5 of a 35-day cycle and prednisone 5 mg twice daily or placebo (n = 315) and prednisone 5 mg twice daily. Primary end points were progression-free survival and overall survival (OS). The secondary end point was time to pain progression (TPP). RESULTS: A 33% reduction (hazard ratio [HR] = 0.67; 95% CI, 0.57 to 0.77; P < .001) was observed in the risk of progression or death with satraplatin versus placebo. This effect was maintained irrespective of prior docetaxel treatment. No difference in OS was seen between the satraplatin and placebo arms (HR = 0.98; 95% CI, 0.84 to 1.15; P = .80). Compared with placebo, satraplatin significantly reduced TPP (HR = 0.64; 95% CI, 0.51 to 0.79; P < .001). Satraplatin was generally well tolerated, although myelosuppression and GI disorders occurred more frequently with satraplatin. CONCLUSION: Oral satraplatin delayed progression of disease and pain in patients with metastatic CRPC experiencing progression after initial chemotherapy but did not provide a significant OS benefit. Satraplatin was generally well tolerated. These results suggest activity for satraplatin in patients with CRPC who experience progression after initial chemotherapy. PMID- 19805693 TI - "Trying to be a good parent" as defined by interviews with parents who made phase I, terminal care, and resuscitation decisions for their children. AB - PURPOSE: When a child's cancer progresses beyond current treatment capability, the parents are likely to participate in noncurative treatment decision making. One factor that helps parents to make these decisions and remain satisfied with them afterward is deciding as they believe a good parent would decide. Because being a good parent to a child with incurable cancer has not been formally defined, we conducted a descriptive study to develop such a definition. METHODS: In face-to-face interviews, 62 parents who had made one of three decisions (enrollment on a phase I study, do not resuscitate status, or terminal care) for 58 patients responded to two open-ended questions about the definition of a good parent and about how clinicians could help them fulfill this role. For semantic content analysis of the interviews, a rater panel trained in this method independently coded all responses. Inter-rater reliability was excellent. RESULTS: Among the aspects of the definition qualitatively identified were making informed, unselfish decisions in the child's best interest, remaining at the child's side, showing the child that he is cherished, teaching the child to make good decisions, advocating for the child with the staff, and promoting the child's health. We also identified 15 clinician strategies that help parents be a part of making these decisions on behalf of a child with advanced cancer. CONCLUSION: The definition and the strategies may be used to guide clinicians in helping parents fulfill the good parent role and take comfort afterward in having acted as a good parent. PMID- 19805695 TI - Influence of NOS1 on verbal intelligence and working memory in both patients with schizophrenia and healthy control subjects. AB - CONTEXT: Human and animal studies have implicated the gene NOS1 in both cognition and schizophrenia susceptibility. OBJECTIVE: To investigate whether a potential schizophrenia risk single-nucleotide polymorphism (rs6490121) identified in a recent genome-wide association study negatively influences cognition in patients with schizophrenia and healthy control subjects. DESIGN: A comparison of both cases and controls grouped according to NOS1 genotype (GG vs AG vs AA) on selected measures of cognition in 2 independent samples. We tested for association between NOS1 rs6490121 and cognitive functions known to be impaired in schizophrenia (IQ, episodic memory, working memory, and attentional control) in an Irish sample. We then sought to replicate the significant results in a German sample. SETTING: Unrelated patients from general adult psychiatric inpatient and outpatient services and unrelated healthy volunteers from the general population were ascertained. PARTICIPANTS: Patients with DSM-IV-diagnosed schizophrenia and healthy control subjects from independent samples of Irish (cases, n = 349; controls, n = 230) and German (cases, n = 232; controls, n = 1344) nationality. RESULTS: A main effect of NOS1 genotype on verbal IQ and working memory was observed in the Irish sample where the homozygous carriers of the schizophrenia risk G allele performed poorly compared with the other genotype groups. These findings were replicated in the German sample, again with the GG genotype carriers performing below other genotype groups. Post hoc analysis of additional IQ measures (full-scale and performance IQ) in the German sample revealed that NOS1 GG carriers underperformed on these measures also. CONCLUSIONS: NOS1 is associated with clinically significant variation in cognition. Whether this is a mechanism by which schizophrenia risk is increased (eg, via an influence on cognitive reserve) is yet to be confirmed. PMID- 19805696 TI - Susceptibility locus on chromosome 1q23-25 for a schizophrenia subtype resembling deficit schizophrenia identified by latent class analysis. AB - CONTEXT: Identifying susceptibility genes for schizophrenia may be complicated by phenotypic heterogeneity, with some evidence suggesting that phenotypic heterogeneity reflects genetic heterogeneity. OBJECTIVE: To evaluate the heritability and conduct genetic linkage analyses of empirically derived, clinically homogeneous schizophrenia subtypes. DESIGN: Latent class and linkage analysis. SETTING: Taiwanese field research centers. PARTICIPANTS: The latent class analysis included 1236 Han Chinese individuals with DSM-IV schizophrenia. These individuals were members of a large affected-sibling-pair sample of schizophrenia (606 ascertained families), original linkage analyses of which detected a maximum logarithm of odds (LOD) of 1.8 (z = 2.88) on chromosome 10q22.3. MAIN OUTCOME MEASURES: Multipoint exponential LOD scores by latent class assignment and parametric heterogeneity LOD scores. RESULTS: Latent class analyses identified 4 classes, with 2 demonstrating familial aggregation. The first (LC2) described a group with severe negative symptoms, disorganization, and pronounced functional impairment, resembling "deficit schizophrenia." The second (LC3) described a group with minimal functional impairment, mild or absent negative symptoms, and low disorganization. Using the negative/deficit subtype, we detected genome-wide significant linkage to 1q23-25 (LOD = 3.78, empiric genome-wide P = .01). This region was not detected using the DSM-IV schizophrenia diagnosis, but has been strongly implicated in schizophrenia pathogenesis by previous linkage and association studies.Variants in the 1q region may specifically increase risk for a negative/deficit schizophrenia subtype. Alternatively, these results may reflect increased familiality/heritability of the negative class, the presence of multiple 1q schizophrenia risk genes, or a pleiotropic 1q risk locus or loci, with stronger genotype-phenotype correlation with negative/deficit symptoms. Using the second familial latent class, we identified nominally significant linkage to the original 10q peak region. CONCLUSION: Genetic analyses of heritable, homogeneous phenotypes may improve the power of linkage and association studies of schizophrenia and thus have relevance to the design and analysis of genome-wide association studies. PMID- 19805697 TI - A reverse-translational study of dysfunctional exploration in psychiatric disorders: from mice to men. AB - CONTEXT: Bipolar mania and schizophrenia are recognized as separate disorders but share many commonalities, which raises the question of whether they are the same disorder on different ends of a continuum. The lack of distinct endophenotypes of bipolar mania and schizophrenia has complicated the development of animal models that are specific to these disorders. Exploration is fundamental to survival and is dysregulated in these 2 disorders. Although exploratory behavior in rodents has been widely studied, surprisingly little work has examined this critical function in humans. OBJECTIVES: To quantify the exploratory behavior of individuals with bipolar mania and schizophrenia and to identify distinctive phenotypes of these illnesses. DESIGN: Static group comparison by the use of a novel human open field paradigm, the human Behavioral Pattern Monitor (BPM). SETTING: Psychiatric hospital. PARTICIPANTS: Fifteen patients with bipolar mania and 16 patients with schizophrenia were compared with 26 healthy volunteers in the human BPM. The effects of amphetamine sulfate, the selective dopamine transporter inhibitor GBR12909, and the genetic knockdown of the dopamine transporter were compared with controls in the mouse BPM. MAIN OUTCOME MEASURES: The amount of motor activity, spatial patterns of activity, and exploration of novel stimuli were quantified in both the human and mouse BPMs. RESULTS: Patients with bipolar mania demonstrated a unique exploratory pattern, characterized by high motor activity and increased object exploration. Patients with schizophrenia did not show the expected habituation of motor activity. Selective genetic or pharmacologic inhibition of the dopamine transporter matched the mania phenotype better than the effects of amphetamine, which has been the criterion standard for animal models of mania. CONCLUSIONS: These findings validate the human open field paradigm and identify defining characteristics of bipolar mania that are distinct from those of schizophrenia. This cross-species study of exploration calls into question an accepted animal model of mania and should help to develop more accurate human and animal models, which are essential to the identification of the neurobiological underpinnings of neuropsychiatric disorders. PMID- 19805698 TI - Incremental benefit and cost of telephone care management and telephone psychotherapy for depression in primary care. AB - CONTEXT: Effectiveness of organized depression care programs is well established, but dissemination will depend on the balance of benefits and costs. OBJECTIVES: To estimate the incremental benefit, incremental cost, and net benefit of 2 depression care programs. DESIGN: Randomized trial comparing 2 interventions with continued usual care, conducted between November 2000 and June 2004. SETTING: Seven primary care clinics of a prepaid health care plan in Washington. PARTICIPANTS: Consecutive primary care patients starting antidepressant treatment were invited to a telephone assessment 2 weeks later. Of 634 patients with significant depressive symptoms, 600 consented and were randomized. INTERVENTIONS: The telephone care management intervention included up to 5 outreach calls for monitoring and support, feedback to treating physicians, and care coordination. The care management plus telephone psychotherapy intervention added an 8-session structured cognitive behavioral therapy program with up to 4 additional calls for reinforcement. MAIN OUTCOME MEASURES: Independent, blinded telephone assessments at 1, 3, 6, 9, 12, and 18 months included the Symptom Checklist 90 depression scale. Health services costs were measured using health care plan accounting records. RESULTS: Over 24 months, telephone care management led to a gain of 29 depression-free days (95% confidence interval, -6 to +63) and a $676 increase in outpatient health care costs (95% confidence interval, $596 lower to $1974 higher). The incremental net benefit was negative even if a day free of depression was valued up to $20. Care management plus psychotherapy led to a gain of 46 depression-free days (95% confidence interval, +12 to +80) and a $397 increase in outpatient costs (95% confidence interval, $882 lower to $1725 higher). The incremental net benefit was positive if a day free of depression was valued at $9 or greater. CONCLUSION: Compared with current primary care practice, a structured telephone program including care management and cognitive behavioral psychotherapy has significant clinical benefit with only a modest increase in health services cost. PMID- 19805699 TI - Association of the Mediterranean dietary pattern with the incidence of depression: the Seguimiento Universidad de Navarra/University of Navarra follow up (SUN) cohort. AB - CONTEXT: Adherence to the Mediterranean dietary pattern (MDP) is thought to reduce inflammatory, vascular, and metabolic processes that may be involved in the risk of clinical depression. OBJECTIVE: To assess the association between adherence to the MDP and the incidence of clinical depression. DESIGN: Prospective study that uses a validated 136-item food frequency questionnaire to assess adherence to the MDP. The MDP score positively weighted the consumption of vegetables, fruit and nuts, cereal, legumes, and fish; the monounsaturated- to saturated-fatty-acids ratio; and moderate alcohol consumption, whereas meat or meat products and whole-fat dairy were negatively weighted. SETTING: A dynamic cohort of university graduates (Seguimiento Universidad de Navarra/University of Navarra Follow-up [SUN] Project). PARTICIPANTS: A total of 10 094 initially healthy Spanish participants from the SUN Project participated in the study. Recruitment began on December 21, 1999, and is ongoing. MAIN OUTCOME MEASURE: Participants were classified as having incident depression if they were free of depression and antidepressant medication at baseline and reported a physician made diagnosis of clinical depression and/or antidepressant medication use during follow-up. RESULTS: After a median follow-up of 4.4 years, 480 new cases of depression were identified. The multiple adjusted hazard ratios (95% confidence intervals) of depression for the 4 upper successive categories of adherence to the MDP (taking the category of lowest adherence as reference) were 0.74 (0.57 0.98), 0.66 (0.50-0.86), 0.49 (0.36-0.67), and 0.58 (0.44-0.77) (P for trend <.001). Inverse dose-response relationships were found for fruit and nuts, the monounsaturated- to saturated-fatty-acids ratio, and legumes. CONCLUSIONS: Our results suggest a potential protective role of the MDP with regard to the prevention of depressive disorders; additional longitudinal studies and trials are needed to confirm these findings. PMID- 19805700 TI - Resting metabolic activity in the cingulate cortex and vulnerability to posttraumatic stress disorder. AB - CONTEXT: Recent neuroimaging research has revealed functional abnormalities in the anterior cingulate cortex, amygdala, and hippocampus in individuals with posttraumatic stress disorder (PTSD). OBJECTIVE: To determine whether resting functional abnormalities found in PTSD are acquired characteristics or familial risk factors. DESIGN: Cross-sectional design including identical twins discordant for trauma exposure. SETTING: Academic medical center. PARTICIPANTS: Combat exposed veterans with PTSD (n = 14) and their identical co-twins not exposed to combat (n = 14) as well as combat-exposed veterans without PTSD (n = 19) and their identical co-twins not exposed to combat (n = 19). MAIN OUTCOME MEASURES: We used positron emission tomography and fluorodeoxyglucose 18 to examine resting regional cerebral metabolic rate for glucose (rCMRglu). RESULTS: Veterans with PTSD and their co-twins had significantly higher resting rCMRglu in the dorsal anterior cingulate cortex/midcingulate cortex (dACC/MCC) compared with veterans without PTSD and their co-twins. Resting rCMRglu in the dACC/MCC in unexposed co twins was positively correlated with combat exposure severity, PTSD symptom severity, and alcohol use in their exposed twins. CONCLUSIONS: Enhanced resting metabolic activity in the dACC/MCC appears to represent a familial risk factor for developing PTSD after exposure to psychological trauma. PMID- 19805701 TI - Improving clinical outcomes in treating heroin dependence: randomized, controlled trial of oral or implant naltrexone. AB - CONTEXT: Oral naltrexone hydrochloride effectively antagonizes heroin, but its utility is limited by patient noncompliance. Sustained-release preparations may overcome this limitation. OBJECTIVE: To compare the safety and efficacy of a single-treatment sustained-release naltrexone implant with daily oral naltrexone treatment. DESIGN: Seventy heroin-dependent volunteers entered a randomized, double-blind, double-placebo controlled trial with a 6-month follow-up period. PATIENTS: Eligibility criteria were DSM-IV opioid (heroin) dependence; age 18 years or older; willingness to be randomized; residing in the Perth, Western Australia, metropolitan area; and completion of preclinical screening and written consent. A total of 129 eligible participants were identified, and 70 (54%) provided informed consent and were randomized as per the study design. INTERVENTION: Participants received oral naltrexone, 50 mg/d, for 6 months (plus placebo implants) or a single dose of 2.3 g of naltrexone implant (plus placebo tablets). MAIN OUTCOME MEASURES: (1) Maintaining therapeutic naltrexone levels above 2 ng/mL; (2) return to regular heroin use (>or=4 d/wk); (3) other heroin use and abstinence; (4) use of illicit nonopioid drugs; (5) number of opiate overdoses requiring hospitalization; (6) treatment-related unexpected and expected adverse events; and (7) blood naltrexone levels (ie, pharmacokinetic profile) for recipients of active naltrexone implants. RESULTS: More participants in the oral vs the implant group had blood naltrexone levels below 2 ng/mL in months 1 (P < .001) and 2 (P = .01); in addition, more oral group participants had returned to regular heroin use by 6 months (P = .003) and at an earlier stage (median [SE], 115 [12.0] days vs 158 [9.4] days). There were 10 trial-related, unexpected adverse events. One serious adverse event, a wound hematoma, was associated with surgical implantation. Naltrexone blood levels in implant recipients were maintained above 1 and 2 ng/mL for 101 (95% confidence interval, 83-119) and 56 (39-73) days, respectively, among men and 124 (88-175) and 43 (16 79) days among women. CONCLUSIONS: The naltrexone implant effectively reduced relapse to regular heroin use compared with oral naltrexone and was not associated with major adverse events. Clinical Trial Registration anzctr.org.au Identifier: ACTRN12606000308594. PMID- 19805702 TI - Cocaine vaccine for the treatment of cocaine dependence in methadone-maintained patients: a randomized, double-blind, placebo-controlled efficacy trial. AB - CONTEXT: Cocaine dependence, which affects 2.5 million Americans annually, has no US Food and Drug Administration-approved pharmacotherapy. OBJECTIVES: To evaluate the immunogenicity, safety, and efficacy of a novel cocaine vaccine to treat cocaine dependence. DESIGN: A 24-week, phase 2b, randomized, double-blind, placebo-controlled trial with efficacy assessed during weeks 8 to 20 and follow up to week 24. SETTING: Cocaine- and opioid-dependent persons recruited from October 2003 to April 2005 from greater New Haven, Connecticut. PARTICIPANTS: One hundred fifteen methadone-maintained subjects (67% male, 87% white, aged 18-46 years) were randomized to vaccine or placebo, and 94 subjects (82%) completed the trial. Most smoked crack cocaine along with using marijuana (18%), alcohol (10%), and nonprescription opioids (44%). INTERVENTION: Over 12 weeks, 109 of 115 subjects received 5 vaccinations of placebo or succinylnorcocaine linked to recombinant cholera toxin B-subunit protein. Main Outcome Measure Semiquantitative urinary cocaine metabolite levels measured thrice weekly with a positive cutoff of 300 ng/mL. RESULTS: The 21 vaccinated subjects (38%) who attained serum IgG anticocaine antibody levels of 43 microg/mL or higher (ie, high IgG level) had significantly more cocaine-free urine samples than those with levels less than 43 microg/mL (ie, low IgG level) and the placebo-receiving subjects during weeks 9 to 16 (45% vs 35% cocaine-free urine samples, respectively). The proportion of subjects having a 50% reduction in cocaine use was significantly greater in the subjects with a high IgG level than in subjects with a low IgG level (53% of subjects vs 23% of subjects, respectively) (P = .048). The most common adverse effects were injection site induration and tenderness. There were no treatment-related serious adverse events, withdrawals, or deaths. CONCLUSIONS: Attaining high (>or=43 microg/mL) IgG anticocaine antibody levels was associated with significantly reduced cocaine use, but only 38% of the vaccinated subjects attained these IgG levels and they had only 2 months of adequate cocaine blockade. Thus, we need improved vaccines and boosters. Trial Registration clinicaltrials.gov Identifier: NCT00142857. PMID- 19805704 TI - Dopaminergic haplotype as a predictor of spatial inattention in children with attention-deficit/hyperactivity disorder. AB - CONTEXT: A distinct pattern of selective attention deficits in attention deficit/hyperactivity disorder (ADHD) has been difficult to identify. Heterogeneity may reflect differences in underlying genetics. OBJECTIVE: To document an objective deficit of selective attention in a large sample of children with and without ADHD using spatial orienting paradigms. By stratifying samples according to the gene dosage of a risk haplotype of the dopamine transporter gene (DAT1), we could determine whether genetic factors predict spatial inattention in ADHD. DESIGN: A case-control design was used. SETTING: Children with ADHD were recruited from clinics or support groups in Ireland. Typically developing children were recruited from schools in and around Dublin, Ireland. PARTICIPANTS: One hundred fifteen children were recruited (ADHD = 50, control = 65). Groups were matched for age but differed in estimated intelligence. INTERVENTION: Two versions of a visual spatial orienting task in which attention was directed by valid, neutral, or invalid cues to target locations. Sudden-onset peripheral cues (exogenous) and centrally presented predictive cues (endogenous) were used. MAIN OUTCOME MEASURES: To isolate an attention deficit in ADHD, groups were first compared using analysis of variance on the spatial orienting tasks. Multiple regression was used to assess the main effect of DAT1 haplotype status (heterozygous vs homozygous) and the interaction of diagnosis and genotype on those variables that discriminated children with and without ADHD. RESULTS: Children with ADHD displayed deficits in reorienting attention from invalidly cued spatial locations, particularly for targets in the left visual field. DAT1 haplotype status predicted spatial reorienting deficits for left visual field targets (P = .007) but there was also a significant interaction of diagnosis and genotype (P = .02), which revealed the greatest impairment in children with ADHD homozygous for the DAT1 haplotype. CONCLUSION: Heterogeneity in selective attention in ADHD can be explained by a replicated genetic risk factor for ADHD, the 10/3 DAT1 haplotype. PMID- 19805703 TI - Positron emission tomography measures of endogenous opioid neurotransmission and impulsiveness traits in humans. AB - CONTEXT: The endogenous opioid system and opioid mu receptors (mu-receptors) are known to interface environmental events, positive (eg, relevant emotional stimuli) and negative (eg, stressors), with pertinent behavioral responses and to regulate motivated behavior. OBJECTIVE: To examine the degree to which trait impulsiveness (the tendency to act on cravings and urges rather than to delay gratification) is predicted by baseline mu-receptor availability or the response of this system to a standardized, experientially matched stressor. DESIGN, SETTING, AND PATIENTS: Nineteen young healthy male volunteers completed a personality questionnaire (NEO Personality Inventory, Revised) and underwent positron emission tomography scans with the mu-receptor-selective radiotracer carfentanil labeled with carbon 11. Measures of receptor concentrations were obtained at rest and during receipt of an experimentally maintained pain stressor of matched intensity between subjects. MAIN OUTCOME MEASURES: Baseline receptor levels and stress-induced activation of mu-opioid system neurotransmission compared between subjects scoring above and below the population median on the NEO Personality Inventory, Revised, impulsiveness subscale and the orthogonal dimension (deliberation) expected to interact with it. RESULTS: High impulsiveness and low deliberation scores were associated with significantly higher regional mu-receptor concentrations and greater stress-induced endogenous opioid system activation. Effects were obtained in the prefrontal and orbitofrontal cortices, anterior cingulate, thalamus, nucleus accumbens, and basolateral amygdala-all regions involved in motivated behavior and the effects of drugs of abuse. Availability of the mu-receptor and the magnitude of stress induced endogenous opioid activation in these regions accounted for 17% to 49% of the variance in these personality traits. CONCLUSIONS: Individual differences in the function of the endogenous mu-receptor system predict personality traits that confer vulnerability to or resiliency against risky behaviors such as the predisposition to develop substance use disorders. These personality traits are also implicated in psychopathological states (eg, personality disorders) in which variations in the function of this neurotransmitter system also may play a role. PMID- 19805705 TI - Pregnancy outcomes in female childhood and adolescent cancer survivors: a linked cancer-birth registry analysis. AB - OBJECTIVE: To compare birth outcomes among female survivors of childhood and adolescent cancer who subsequently bear children, relative to those of women without a history of cancer. DESIGN: Retrospective cohort study. SETTING: Four US regions. PARTICIPANTS: Cancer registries identified girls younger than 20 years who were diagnosed as having cancer from 1973 through 2000. Linked birth records identified the first live births after diagnosis (n = 1898). Comparison subjects were selected from birth records (n = 14 278). Survivors of genital tract carcinomas underwent separate analysis. MAIN EXPOSURE: Cancer diagnosis at younger than 20 years. MAIN OUTCOME MEASURES: Infant low birth weight, preterm delivery, sex ratio, malformations, mortality, and delivery method, and maternal diabetes, anemia, and preeclampsia. RESULTS: Infants born to childhood cancer survivors were more likely to be preterm (relative risk [RR], 1.54; 95% confidence interval [CI], 1.30-1.83) and to weigh less than 2500 g (1.31; 1.10 1.57). For the offspring of genital tract carcinoma survivors, RRs were 1.33 (95% CI, 1.13-1.56) and 1.29 (1.10-1.53), respectively. There were no increased risks of malformations, infant death, or altered sex ratio, suggesting no increased germ cell mutagenicity. In exploratory analysis, bone cancer survivors had an increased risk of diabetes (RR, 4.92; 95% CI, 1.60-15.13), and anemia was more common among brain tumor survivors (3.05; 1.16-7.98) and childhood cancer survivors whose initial treatment was chemotherapy only (2.45; 1.16-5.17). CONCLUSIONS: Infants born to female survivors of childhood and adolescent cancer were not at increased risk of malformations or death. Increased occurrence of preterm delivery and low birth weight suggest that close monitoring is warranted. Increased diabetes and anemia among subgroups have not been reported, suggesting areas for study. PMID- 19805707 TI - End-of-life decisions in Dutch neonatal intensive care units. AB - OBJECTIVE: To clarify the practice of end-of-life decision making in severely ill newborns. DESIGN: Retrospective descriptive study with face-to-face interviews. SETTING: The 10 neonatal intensive care units in the Netherlands from October 2005 to September 2006. PATIENTS: All 367 newborn infants who died in the first 2 months of life in Dutch neonatal intensive care units. Adequate documentation was available in 359 deaths. OUTCOME MEASURES: Presence of end-of-life decisions, classification of deaths in 3 groups, and physicians' considerations leading to end-of-life decisions. RESULTS: An end-of-life decision preceded death in 95% of cases, and in 5% treatment was continued until death. Of all of the deaths, 58% were classified as having no chance of survival and 42% were stabilized newborns with poor prognoses. Withdrawal of life-sustaining therapy was the main mode of death in both groups. One case of deliberate ending of life was found. In 92% of newborns with poor prognoses, end-of-life decisions were based on patients' future quality of life and mainly concerned future suffering. Considerations regarding the infant's present state were made in 44% of infants. CONCLUSIONS: Virtually all deaths in Dutch neonatal intensive care units are preceded by the decision to withdraw life-sustaining treatment and many decisions are based on future quality of life. The decision to deliberately end the life of a newborn may occur less frequently than was previously assumed. PMID- 19805706 TI - Reproductive outcomes in male childhood cancer survivors: a linked cancer-birth registry analysis. AB - OBJECTIVE: To compare the risk of reproductive and infant outcomes between male childhood cancer survivors and a population-based comparison group. DESIGN: Retrospective cohort study. SETTING: Four US regions. PARTICIPANTS: Cancer registries identified males younger than 20 years diagnosed with cancer from 1973 to 2000. Linked birth certificates identified first subsequent live offspring (N = 470). Comparison subjects were identified from remaining birth certificates, frequency-matched on year and age at fatherhood, and race/ethnicity (N = 4150). MAIN EXPOSURE: Cancer diagnosis before age 20 years. OUTCOME MEASURES: Pregnancy and infant outcomes identified from birth certificates. RESULTS: Compared with infants born to unaffected males, offspring of cancer survivors had a borderline risk of having a birth weight less than 2500 g (relative risk, 1.43 [95% confidence interval, 0.99-2.05]) that was associated most strongly with younger age at cancer diagnosis and exposure to any chemotherapy (1.96 [1.22-3.17]) or radiotherapy (1.95 [1.14-3.35]). However, they were not at risk of being born prematurely, being small for gestational age, having malformations, or having an altered male to female ratio. Overall, female partners of male survivors were not more likely to have maternal complications recorded on birth records vs the comparison group. However, preeclampsia was associated with some cancers, especially central nervous system tumors (relative risk, 3.36 [95% confidence interval, 1.63-6.90]). CONCLUSIONS: Most pregnancies resulting in live births among partners of male childhood cancer survivors were not at significantly greater risk of complications vs comparison subjects. However, there remains the possibility that prior cancer therapy may affect male germ cells with some effects on progeny and on female partners. PMID- 19805708 TI - Intervention at the border of viability: perspective over a decade. AB - OBJECTIVE: To investigate prenatal management and outcome of infants born at the border of viability during 2 periods, 2001 to 2003 (late epoch) and 1993 to 1995 (early epoch). DESIGN: Cohort study. SETTING: Single academic, high-risk perinatal referral center. PARTICIPANTS: All 160 women admitted to labor and delivery with a live fetus who delivered at an estimated gestational age of 220/7 weeks to 246/7 weeks. MAIN OUTCOME MEASURES: Prenatal management and time between maternal admission and delivery or death of the fetus, infant resuscitation efforts, neonatal intensive care unit interventions, time of death, and morbidities in survivors. RESULTS: Mothers in both epochs were of similar age, race, and duration of pregnancy at hospital admission. Compared with the early epoch, women during the late epoch were more likely to be transported to a higher level of care (relative risk [RR], 2.01; 95% confidence interval [CI], 1.58-2.57) and receive sonographic surveillance (RR, 1.48; 95% CI, 1.07-2.04), antibiotics (RR, 1.60; 95% CI, 1.10-2.33), and antenatal steroids (RR, 1.61; 95% CI, 1.22 2.12). Life-sustaining interventions were provided for infants admitted to the neonatal intensive care unit more frequently during the late epoch than the early epoch, including high-frequency ventilation (RR, 3.57; 95% CI, 1.93-6.61), chest tubes (RR, 1.44; 95% CI, 1.06-1.94), dopamine administration (RR, 2.49; 95% CI, 1.24-4.97), and steroid administration for blood pressure support (RR, 2.18; 95% CI, 1.60-2.92). Gestational age-specific mortality was the same in the 2 epochs. CONCLUSIONS: More interventions were provided for infants born at 22 to 24 weeks' gestation in the late epoch than the early epoch. Despite these changes in management, there has been no reduction in mortality in more than a decade. PMID- 19805709 TI - Characteristics and concordance of autism spectrum disorders among 277 twin pairs. AB - OBJECTIVES: To examine patterns of autism spectrum disorder (ASD) inheritance and other features in twin pairs by zygosity, sex, and specific ASD diagnosis. DESIGN: Cross-sectional study. SETTING: Internet-based autism registry for US residents. PARTICIPANTS: Survey results from 277 twin pairs (210 dizygotic [DZ] and 67 monozygotic [MZ]) aged 18 years or younger with at least 1 affected twin. MAIN EXPOSURES: Zygosity and sex. OUTCOME MEASURES: Concordance within twin pairs of diagnosis, natural history, and results from standardized autism screening. RESULTS: Pairwise ASD concordance was 31% for DZ and 88% for MZ twins. Female and male MZ twins were 100% and 86% concordant, respectively, and DZ twin pairs with at least 1 female were less likely to be concordant (20%) than were male-male DZ twin pairs (40%). The hazard ratio for ASD diagnosis of the second twin after a first-twin diagnosis was 7.48 for MZ vs DZ twins (95% confidence interval, 3.8 14.7). Affected DZ individual twins had an earlier age at first parental concern and more frequent diagnoses of intellectual disability than did MZ twins; MZ twins had a higher prevalence of bipolar disorder and Asperger syndrome and higher concordance of the latter. Results of autism screening correlated with parent-reported ASD status in more than 90% of cases. CONCLUSIONS: Our data support greater ASD concordance in MZ vs DZ twins. Overall higher functioning, psychiatric comorbidity, and Asperger syndrome concordance among affected MZ vs DZ twins may also suggest differential heritability for different ASDs. For families in which one MZ twin is diagnosed with ASD, the second twin is unlikely to receive an ASD diagnosis after 12 months. In addition, Internet parent report of ASD status is valid. PMID- 19805710 TI - Randomized clinical trial of behavioral intervention and nutrition education to improve caloric intake and weight in children with cystic fibrosis. AB - OBJECTIVE: To evaluate the efficacy of a behavioral plus nutrition education intervention, Be In CHARGE!, compared with that of a nutrition education intervention alone on caloric intake and weight gain in children with cystic fibrosis and pancreatic insufficiency. DESIGN: Randomized controlled trial. SETTING: Cystic fibrosis centers in the eastern, midwestern, and southern United States. PARTICIPANTS: Seventy-nine children aged 4 to 12 years below the 40th percentile for weight for age were recruited. Sixty-seven completed the intervention and 59 completed a 24-month follow-up assessment. INTERVENTION: Comparison of a behavioral plus nutrition education intervention with a nutrition education intervention alone. MAIN OUTCOME MEASURES: Primary outcomes were changes from pretreatment to posttreatment in caloric intake and weight gain. Secondary outcomes were changes from pretreatment to posttreatment in percentage of the estimated energy requirement and body mass index z score. These outcomes were also examined 24 months posttreatment. RESULTS: After treatment, the behavioral plus nutrition education intervention as compared with the nutrition education intervention alone had a statistically greater average increase on the primary and secondary outcomes of caloric intake (mean, 872 vs 489 cal/d, respectively), percentage of the estimated energy requirement (mean, 148% vs 127%, respectively), weight gain (mean, 1.47 vs 0.92 kg, respectively), and body mass index z score (0.38 vs 0.18, respectively). At the 24-month follow-up, children in both conditions maintained an estimated energy requirement of around 120% and did not significantly differ on any outcomes. CONCLUSIONS: A behavioral plus nutrition education intervention was more effective than a nutrition education intervention alone at increasing dietary intake and weight over a 9 week period. However, across the 24-month follow-up, both treatments achieved similar outcomes. Trial Registration clinicaltrials.gov Identifier: NCT00006169. PMID- 19805711 TI - A longitudinal study of maternal depression and child maltreatment in a national sample of families investigated by child protective services. AB - OBJECTIVE: To assess whether a change in depression predicts a mother's change in maltreatment. DESIGN: Observational, repeated measures study. SETTING: National Survey of Child and Adolescent Well-being, 1999 to 2004. PARTICIPANTS: Mothers who retained custody of a child aged 0 to 15 years following a maltreatment investigation and completed at least 2 of 3 surveys (n = 2386). MAIN EXPOSURE: Change in depression status between baseline and 18- and 36-month follow-ups, assessed with the Composite International Diagnostic Interview Short Form. MAIN OUTCOME MEASURES: Change in psychological aggression, physical assault, and neglect between baseline and 18- and 36-month follow-ups, assessed with the Conflict Tactics Scale Parent-Child version. RESULTS: One-third (35.5%) of mothers experienced onset or remission of depression. Onset of depression was associated with an increase of 2.3 (95% confidence interval, 0.2-4.4) psychologically aggressive acts in an average 12-month period, but was not statistically significantly associated with change in physical assault or neglect. CONCLUSION: Depression is positively associated with maternal perpetration of psychological aggression in high-risk families. PMID- 19805712 TI - Violence against women and increases in the risk of diarrheal disease and respiratory tract infections in infancy: a prospective cohort study in Bangladesh. AB - OBJECTIVE: To explore whether different forms of violence against women were associated with increased incidence rates of diarrhea and respiratory tract infections among infants. DESIGN: A 12-month follow-up study embedded in a food and micronutrient supplementation trial. SETTING: Rural Bangladesh. PARTICIPANTS: Pregnant women and their 3132 live-born children. MAIN EXPOSURE: Maternal exposure to physical, sexual, and emotional violence and level of controlling behavior in the family. MAIN OUTCOME MEASURES: Infants' risk of falling ill with diarrheal diseases and respiratory tract infections in relation to mothers' exposure to different forms of violence. Adjusted for household economic conditions, mother's education level, parity, and religion. RESULTS: Fifty percent of the women reported lifetime experience of family violence. Infants of mothers exposed to different forms of family violence had 26% to 37% higher incidence of diarrhea. Any lifetime family violence was positively associated with increased incidence of diarrheal diseases (adjusted rate ratio, 1.20; 95% confidence interval, 1.10-1.30) and lower respiratory tract infections (adjusted rate ratio, 1.31; 95% confidence interval, 1.17-1.46). Further, all forms of family violence were also independently positively associated with infant illness, and the highest incidence rates were found among the daughters of severely physically abused mothers. CONCLUSION: Family violence against women was positively associated with an increased risk of falling ill with diarrheal and respiratory tract infections during infancy. The present findings add to increasing evidence of the magnitude of public health consequences of violence against women. PMID- 19805713 TI - Predictive values of psychiatric symptoms for internet addiction in adolescents: a 2-year prospective study. AB - OBJECTIVES: To evaluate the predictive values of psychiatric symptoms for the occurrence of Internet addiction and to determine the sex differences in the predictive value of psychiatric symptoms for the occurrence of Internet addiction in adolescents. DESIGN: Internet addiction, depression, attention deficit/hyperactivity disorder, social phobia, and hostility were assessed by self-reported questionnaires. Participants were then invited to be assessed for Internet addiction 6, 12, and 24 months later (the second, third, and fourth assessments, respectively). SETTING: Ten junior high schools in southern Taiwan. PARTICIPANTS: A total of 2293 (1179 boys and 1114 girls) adolescents participated in the initial investigation. MAIN EXPOSURE: The course of time. MAIN OUTCOME MEASURE: Internet addiction as assessed using the Chen Internet Addiction Scale. RESULTS: Depression, attention-deficit/hyperactivity disorder, social phobia, and hostility were found to predict the occurrence of Internet addiction in the 2 year follow-up, and hostility and attention-deficit/hyperactivity disorder were the most significant predictors of Internet addiction in male and female adolescents, respectively. CONCLUSIONS: These results suggest that attention deficit/hyperactivity disorder, hostility, depression, and social phobia should be detected early on and intervention carried out to prevent Internet addiction in adolescents. Also, sex differences in psychiatric comorbidity should be taken into consideration when developing prevention and intervention strategies for Internet addiction. PMID- 19805714 TI - Impact of individual values on adherence to emergency contraception practice guidelines among pediatric residents: implications for training. AB - OBJECTIVE: To evaluate the impact of individual, system, and interpersonal factors on emergency contraception practices. We hypothesized that abortion attitudes and attitudes toward teen sex would be significant individual factors influencing emergency contraception practices. DESIGN: This was a cross sectional, anonymous Internet survey. SETTING: Four pediatric residency programs in the Baltimore, Maryland-Washington, DC, metropolitan area during April to June 2007. PARTICIPANTS: One hundred forty-one pediatric residents completed the survey. MAIN EXPOSURE: Abortion attitudes were assessed by participants' level of agreement with abortion in 7 scenarios. Attitudes toward teen sex were assessed by participants' level of agreement with 5 statements about the acceptability of teens having sex. MAIN OUTCOME MEASURES: Emergency contraceptive counseling behavior was assessed by reported frequency of including emergency contraception in routine contraceptive counseling. Intention to prescribe emergency contraception was assessed by reported likelihood of prescribing in 5 scenarios. RESULTS: When controlling for demographics and other predictors, residents with less favorable abortion attitudes were more likely to have the lowest intention to prescribe emergency contraception. Residents with more positive attitudes toward teen sex and who had a preceptor encourage emergency contraception prescription were more likely to include emergency contraception in routine contraceptive counseling most/all the time and to have the highest intention to prescribe. CONCLUSION: Efforts to challenge and affect attitudes toward teen sex and to prompt residents to prescribe emergency contraception in clinical settings may be needed to encourage more proactive emergency contraceptive practice in accordance with national practice guidelines. PMID- 19805715 TI - Selective serotonin reuptake inhibitor exposure in utero and pregnancy outcomes. AB - OBJECTIVE: To investigate the effect of intrauterine selective serotonin reuptake inhibitor (SSRI) exposure on pregnancy outcomes. DESIGN: Prospective cohort study. SETTING: Department of Obstetrics, Aarhus University Hospital, Aarhus, Denmark. PARTICIPANTS: Pregnant women receiving prenatal care in our hospital from 1989 to 2006. MAIN EXPOSURE: Maternal SSRI use during pregnancy. OUTCOME MEASURES: Gestational age, birth weight, head circumference, 5-minute Apgar score, and admission to the neonatal intensive care unit. RESULTS: Three hundred twenty-nine pregnant women reported treatment with SSRIs, 4902 were not treated with SSRIs but had a history of psychiatric illness, and 51 770 reported no history of psychiatric illness. Gestational age was 5 days (95% confidence interval [CI], -6 to -3) shorter and the odds ratio (OR) for preterm birth was 2.0 (95% CI, 1.3-3.2) in the women exposed to SSRIs compared with women with no history of psychiatric illness. In utero-exposed newborns had increased risk of admission to the neonatal intensive care unit (OR, 2.4; 95% CI, 1.7-3.4) and of 5 minute Apgar scores of less than 8 (OR, 4.4; 95% CI, 2.6-7.6) compared with those not exposed. Head circumference and birth weight did not differ between infants in the exposed and unexposed groups. The results were similar when compared with infants of women with a psychiatric history. CONCLUSIONS: Exposure to SSRIs during pregnancy was associated with an increased risk of preterm delivery, a low 5-minute Apgar score, and neonatal intensive care unit admission, which was not explained by lower Apgar scores or gestational age. The study justifies increased awareness to the possible effects of intrauterine exposure to antidepressants. PMID- 19805716 TI - Picture of the month. Eyelid pilomatricoma. PMID- 19805718 TI - Death in the Netherlands: evidence and argument. PMID- 19805719 TI - Trapped in the net: will internet addiction become a 21st-century epidemic? PMID- 19805720 TI - Formal swimming lessons must be defined. PMID- 19805721 TI - Not one more child drowns. PMID- 19805722 TI - Internet safety. PMID- 19805723 TI - Clinical reasoning: A 54-year-old woman with hand dysesthesia: many dimensions to a common problem. PMID- 19805724 TI - Teaching video neuroimages: excessive grinning in Wilson disease. PMID- 19805726 TI - Driving under low-contrast visibility conditions in Parkinson disease. AB - OBJECTIVE: To assess driving performance in Parkinson disease (PD) under low contrast visibility conditions. METHODS: Licensed, active drivers with mild to moderate PD (n = 67, aged 66.2 +/- 9.0 years, median Hoehn-Yahr stage = 2) and controls (n = 51, aged 64.0 +/- 7.2 years) drove in a driving simulator under high- (clear sky) and low-contrast visibility (fog) conditions, leading up to an intersection where an incurring vehicle posed a crash risk in fog. RESULTS: Drivers with PD had higher SD of lateral position (SDLP) and lane violation counts (LVC) than controls during fog (p < 0.001). Transition from high- to low contrast visibility condition increased SDLP and LVC more in PD than in controls (p < 0.01). A larger proportion of drivers with PD crashed at the intersection in fog (76.1% vs 37.3%, p < 0.0001). The time to first reaction in response to incursion was longer in drivers with PD compared with controls (median 2.5 vs 2.0 seconds, p < 0.0001). Within the PD group, the strongest predictors of poor driving outcomes under low-contrast visibility conditions were worse scores on measures of visual processing speed and attention, motion perception, contrast sensitivity, visuospatial construction, motor speed, and activities of daily living score. CONCLUSIONS: During driving simulation under low-contrast visibility conditions, drivers with Parkinson disease (PD) had poorer vehicle control and were at higher risk for crashes, which were primarily predicted by decreased visual perception and cognition; motor dysfunction also contributed. Our results suggest that drivers with PD may be at risk for unsafe driving in low contrast visibility conditions such as during fog or twilight. PMID- 19805727 TI - SPG15 is the second most common cause of hereditary spastic paraplegia with thin corpus callosum. AB - OBJECTIVE: Hereditary spastic paraplegias (HSPs) are very heterogeneous inherited neurodegenerative disorders. Our group recently identified ZFYVE26 as the gene responsible for one of the clinical and genetic entities, SPG15. Our aim was to describe its clinical and mutational spectra. METHODS: We analyzed all exons of SPG15/ZFYVE26 gene by direct sequencing in a series of 60 non-SPG11 HSP subjects with associated mental or MRI abnormalities, including 30 isolated cases. The clinical data were collected through the SPATAX network. RESULTS: We identified 13 novel truncating mutations in ZFYVE26, 12 of which segregated at the homozygous or compound heterozygous states in 8 new SPG15 families while 1 was found at the heterozygous state in a single family. Two of 3 splice site mutations were validated on mRNA of 2 patients. The SPG15 phenotype in 11 affected individuals was characterized by early onset HSP, severe progression of the disease, and mental impairment dominated by cognitive decline. Thin corpus callosum and white matter hyperintensities were MRI hallmarks of the disease in this series. CONCLUSIONS: The mutations are truncating, private, and distributed along the entire coding sequence of ZFYVE26, which complicates the analysis of this gene in clinical practice. In our series of patients with hereditary spastic paraplegia-thin corpus callosum, the largest analyzed so far, SPG15 was the second most frequent form (11.5%) after SPG11. Both forms share similar clinical and imaging presentations with very few distinctions, which are, however, insufficient to infer the molecular diagnosis when faced with a single patient. PMID- 19805728 TI - Emergence from minimally conscious state: insights from evaluation of posttraumatic confusion. AB - BACKGROUND: Guidelines for defining the minimally conscious state (MCS) specify behaviors that characterize emergence, including "reliable and consistent" functional communication (accurate yes/no responding). Guidelines were developed by consensus because of lack of empirical data. OBJECTIVE: To evaluate the utility of the operational threshold for emergence from posttraumatic MCS, by determining yes/no accuracy to questions of varied difficulty, including simple orientation questions, using all items from the Yes/No Subscale of the Mississippi Aphasia Screening Test. METHOD: Prospective observational study of a cohort of responsive patients recovering from traumatic brain injury in an acute inpatient brain injury rehabilitation program. RESULTS: Of the 629 observations from 144 participants, name recognition was the easiest yes/no question, with nonconfused individuals responding with 100% accuracy, whereas only 75% to 78% of confused participants on initial evaluation answered this question correctly. Generalized Estimating Equations analysis revealed that confused participants were more likely to respond inaccurately to all yes/no questions. Nonconfused participants had a reduction in odds of inaccuracy ranging from 45.6% to 99.7% (p = 0.001 to 0.02) depending on the type of yes/no question. CONCLUSIONS: Accuracy for simple orientation yes/no questions remains challenging for responsive patients in early recovery from traumatic brain injury. Although name recognition questions are relatively easier than other types of yes/no questions, including situational orientation questions, confused patients still may answer these incorrectly. Results suggest the operational threshold for yes/no response accuracy as a diagnostic criterion for emergence from the minimally conscious state should be revisited, with particular consideration of the type of yes/no questions and the requisite accuracy threshold for responses. PMID- 19805729 TI - Relative preservation of MMSE scores in autopsy-proven dementia with Lewy bodies. AB - BACKGROUND: Recent studies raised questions about the severity of cognitive impairment associated with dementia with Lewy bodies (DLB). However, there have been few analyses of large, multicenter data registries for clinical-pathologic correlation. METHODS: We evaluated data from the National Alzheimer's Coordinating Center registry (n = 5,813 cases meeting initial inclusion criteria) and the University of Kentucky Alzheimer's Disease Center autopsy series (n = 527) to compare quantitatively the severity of cognitive impairment associated with DLB pathology vs Alzheimer disease (AD) and AD+DLB pathologies. RESULTS: Mini-Mental State Examination (MMSE) scores showed that persons with pure DLB had cognitive impairment of relatively moderate severity (final MMSE score 15.6 +/- 8.7) compared to patients with pure AD and AD+DLB (final MMSE score 10.7 +/- 8.6 and 10.6 +/- 8.6). Persons with pure DLB pathology from both data sets had more years of formal education and were more likely to be male. Differences in final MMSE scores were significant (p < 0.01) between pure DLB and both AD+DLB and pure AD even after correction for education level, gender, and MMSE-death interval. Even in cases with extensive neocortical LBs, the degree of cognitive impairment was most strongly related to the amount of concomitant AD-type neurofibrillary pathology. CONCLUSIONS: Dementia with Lewy bodies can constitute a debilitating disease with associated psychiatric, motoric, and autonomic dysfunction. However, neocortical Lewy bodies are not a substrate for severe global cognitive impairment as assessed by the Mini-Mental State Examination. Instead, neocortical Lewy bodies appear to constitute or reflect an additive disease process, requiring Alzheimer disease or other concomitant brain diseases to induce severe global cognitive deterioration. PMID- 19805730 TI - The video head impulse test: diagnostic accuracy in peripheral vestibulopathy. AB - BACKGROUND: The head impulse test (HIT) is a useful bedside test to identify peripheral vestibular deficits. However, such a deficit of the vestibulo-ocular reflex (VOR) may not be diagnosed because corrective saccades cannot always be detected by simple observation. The scleral search coil technique is the gold standard for HIT measurements, but it is not practical for routine testing or for acute patients, because they are required to wear an uncomfortable contact lens. OBJECTIVE: To develop an easy-to-use video HIT system (vHIT) as a clinical tool for identifying peripheral vestibular deficits. To validate the diagnostic accuracy of vHIT by simultaneous measures with video and search coil recordings across healthy subjects and patients with a wide range of previously identified peripheral vestibular deficits. METHODS: Horizontal HIT was recorded simultaneously with vHIT (250 Hz) and search coils (1,000 Hz) in 8 normal subjects, 6 patients with vestibular neuritis, 1 patient after unilateral intratympanic gentamicin, and 1 patient with bilateral gentamicin vestibulotoxicity. RESULTS: Simultaneous video and search coil recordings of eye movements were closely comparable (average concordance correlation coefficient r(c) = 0.930). Mean VOR gains measured with search coils and video were not significantly different in normal (p = 0.107) and patients (p = 0.073). With these groups, the sensitivity and specificity of both the reference and index test were 1.0 (95% confidence interval 0.69-1.0). vHIT measures detected both overt and covert saccades as accurately as coils. CONCLUSIONS: The video head impulse test is equivalent to search coils in identifying peripheral vestibular deficits but easier to use in clinics, even in patients with acute vestibular neuritis. PMID- 19805731 TI - Intraepidermal nerve fiber density and its application in sarcoidosis. AB - BACKGROUND: Intraepidermal nerve fiber density (IENFD) is considered a good diagnostic tool for small fiber neuropathy (SFN). OBJECTIVES: To assess stratified normative values for IENFD and determine the reliability and validity of IENFD in sarcoidosis. METHODS: IENFD was assessed in 188 healthy volunteers and 72 patients with sarcoidosis (n = 58 with SFN symptoms, n = 14 without SFN symptoms). Healthy controls were stratified (for age and sex), resulting in 6 age groups (20-29, 30-39, ... up to > or = 70 years) containing at least 15 men and 15 women. A skin biopsy was taken in each participant 10 cm above the lateral malleolus and analyzed in accordance with the international guidelines using bright-field microscopy. Interobserver/intraobserver reliability of IENFD was examined. In the patients, a symptoms inventory questionnaire (SIQ; assessing SFN symptoms) and the Vickrey Peripheral Neuropathy Quality-of-Life Instrument-97 (PNQoL-97) were assessed to examine the discriminative ability of normative IENFD values. RESULTS: There was a significant age-dependent decrease of IENFD values in healthy controls, with lower densities in men compared with women. Good interobserver/intraobserver reliability scores were obtained (kappa values > or = 0.90). A total of 21 patients with sarcoidosis had a reduced IENFD score (< 5th percentile; 19 [32.8%] in patients with SFN symptoms, 2 [14.3%] in patients without SFN symptoms). The validity of the normative IENFD values was demonstrated by distinguishing between the SIQ scores and various PNQoL-97 values for the different patient groups. CONCLUSION: This study provides clinically applicable distal intraepidermal nerve fiber density normative values, showing age- and sex-related differences. PMID- 19805732 TI - Gamma knife radiosurgery for multiple sclerosis-related trigeminal neuralgia. AB - BACKGROUND: Surgical options for multiple sclerosis (MS) related to trigeminal neuralgia (TN), a severe and disabling pain disorder, include percutaneous rhizotomy, stereotactic radiosurgery, or microsurgical nerve section. Our goal was to evaluate clinical outcomes after gamma knife radiosurgery (GKRS) in patients with MS with TN. METHODS: We evaluated clinical outcomes in 37 patients with TN managed over a 12-year period. The maximum TN target dose varied between 70 and 90 Gy. Seventy-eight percent of patients had failed prior surgery. In 9, GKRS was the first procedure. Median follow-up was 56.7 months (range, 6-174). Pain relief was assessed in each patient by physicians who did not participate in the surgery. RESULTS: Eventual complete pain relief (BNI grade I) after GKRS and reasonable pain control (BNI grade I-IIIb) after GKRS were noted in 23 patients (62.1%) and 36 patients (97.3%) at some point in their course. Reasonable pain control (BNI grade I-IIIb) after GKRS was maintained in 82.6%, 73.9%, and 54.0% of patients after 1, 3, and 5 years. Fourteen patients (37.8%) underwent a second or a subsequent procedure for residual or recurrent pain. Eight patients underwent a second GKRS, 5 underwent percutaneous glycerol rhizotomy, and 1 underwent balloon microcompression. The complication rate after GKRS was 5.4% (new onset of nondisabling paresthesias). No patient developed dysesthesias. CONCLUSIONS: Gamma knife radiosurgery is the most minimally invasive surgical technique for multiple sclerosis-related trigeminal neuralgia and has low morbidity. For this reason, gamma knife radiosurgery proved to be a satisfactory management strategy for multiple sclerosis-related trigeminal neuralgia. PMID- 19805733 TI - Hughlings Jackson's suggestion for the treatment of epilepsy. AB - John Hughlings Jackson articulated a neurologic method of systematically evaluating the anatomy, physiology, and pathology of every patient with neurologic disease. He used this mode of analysis to develop a theory of the physiology of epilepsy. We examined an example of his method in a newly discovered, unpublished manuscript containing his suggestions for the treatment of epilepsy based on his physiologic ideas. He had his private papers destroyed at the time of his death, but the Rockefeller Library of the University College London Institute of Neurology, Queen Square, contains a collection of his papers probably saved from destruction by his collaborator James Taylor. Among these articles is an 1899 memorandum, labeled "For Private Circulation" and entitled "A Suggestion for the Treatment of Epilepsy." In it, Hughlings Jackson claimed that focal discharging lesions cause both focal and generalized epilepsy, and that the cells in the lesion discharge their energy more easily than normal tissue. Citing microscopic evidence that such lesions are congested and inflamed, and that tuberculin destroys such tissue in the lung, he reasoned that destroying these unstable neurons with tuberculin would improve epilepsy. In this private manuscript, Hughlings Jackson uses an unusually detailed analysis of the pathology, anatomy, and physiology of epilepsy to predict a scientific approach to its treatment. PMID- 19805734 TI - Core-rod myopathy caused by mutations in the nebulin gene. PMID- 19805735 TI - LINGO1 variant increases risk of familial essential tremor. PMID- 19805736 TI - Ageotropic central positional nystagmus in nodular infarction. PMID- 19805737 TI - Cobblestone-like brain dysgenesis and altered glycosylation in congenital cutis laxa, Debre type. PMID- 19805738 TI - Neurologic improvement after peripheral blood stem cell transplantation in poems. PMID- 19805739 TI - Cell-cell and intracellular lactate shuttles. AB - Once thought to be the consequence of oxygen lack in contracting skeletal muscle, the glycolytic product lactate is formed and utilized continuously in diverse cells under fully aerobic conditions. 'Cell-cell' and 'intracellular lactate shuttle' concepts describe the roles of lactate in delivery of oxidative and gluconeogenic substrates as well as in cell signalling. Examples of the cell-cell shuttles include lactate exchanges between between white-glycolytic and red oxidative fibres within a working muscle bed, and between working skeletal muscle and heart, brain, liver and kidneys. Examples of intracellular lactate shuttles include lactate uptake by mitochondria and pyruvate for lactate exchange in peroxisomes. Lactate for pyruvate exchanges affect cell redox state, and by itself lactate is a ROS generator. In vivo, lactate is a preferred substrate and high blood lactate levels down-regulate the use of glucose and free fatty acids (FFA). As well, lactate binding may affect metabolic regulation, for instance binding to G-protein receptors in adipocytes inhibiting lipolysis, and thus decreasing plasma FFA availability. In vitro lactate accumulation upregulates expression of MCT1 and genes coding for other components of the mitochondrial reticulum in skeletal muscle. The mitochondrial reticulum in muscle and mitochondrial networks in other aerobic tissues function to establish concentration and proton gradients necessary for cells with high mitochondrial densities to oxidize lactate. The presence of lactate shuttles gives rise to the realization that glycolytic and oxidative pathways should be viewed as linked, as opposed to alternative, processes, because lactate, the product of one pathway, is the substrate for the other. PMID- 19805740 TI - Influence of endogenous angiotensin II on control of sympathetic nerve activity in human dehydration. AB - Arterial blood pressure can often fall too low during dehydration, leading to an increased incidence of orthostatic hypotension and syncope. Systemic sympathoexcitation and increases in volume regulatory hormones such as angiotensin II (AngII) may help to maintain arterial pressure in the face of decreased plasma volume. Our goals in the present study were to quantify muscle sympathetic nerve activity (MSNA) during dehydration (DEH), and to test the hypothesis that endogenous increases in AngII in DEH have a mechanistic role in DEH-associated sympathoexcitation. We studied 17 subjects on two separate study days: DEH induced by 24 h fluid restriction and a euhydrated (EUH) control day. MSNA was measured by microneurography at the peroneal nerve, and arterial blood pressure, electrocardiogram, and central venous pressure were also recorded continuously. Sequential nitroprusside and phenylephrine (modified Oxford test) were used to evaluate baroreflex control of MSNA. Losartan (angiotensin type 1 receptor (AT1) antagonist) was then administered and measurements were repeated. MSNA was elevated during DEH (42 +/- 5 vs. EUH: 32 +/- 4 bursts per 100 heartbeats, P = 0.02). Blockade of AT1 receptors partially reversed this change in MSNA during DEH while having no effect in the control EUH condition. The sensitivity of baroreflex control of MSNA was unchanged during DEH compared to EUH. We conclude that endogenous increases in AngII during DEH contribute to DEH associated sympathoexcitation. PMID- 19805741 TI - Extracellular ATP inhibits chloride channels in mature mammalian skeletal muscle by activating P2Y1 receptors. AB - ATP is released from skeletal muscle during exercise, a discovery dating back to 1969. Surprisingly, few studies have examined the effects of extracellular ATP on mature mammalian skeletal muscle. This electrophysiological study examined the effects of extracellular ATP on fully innervated rat levator auris longus using two intracellular microelectrodes. The effects of ATP were determined by measuring the relative changes of miniature endplate potentials (mEPPs) and voltage responses to step current pulses in individual muscle fibres. Exposure to ATP (20 microm) prolonged the mEPP falling phase by 31 +/- 7.5% (values +/- s.d., n = 3 fibres). Concurrently, the input resistance increased by 31 +/- 2.0% and the time course of the voltage responses increased by 59 +/- 3.0%. Analogous effects were observed using 2 and 5 microm ATP, and on regions distal from the neuromuscular junction, indicating that physiologically relevant levels of ATP enhanced electrical signalling over the entire muscle fibre. The effects of extracellular ATP were blocked by 200 microm anthracene-9-carboxylic acid, a chloride channel inhibitor, and reduced concentrations of extracellular chloride, indicating that ATP inhibited chloride channels. A high affinity agonist for P2Y receptors, 2-methylthioadenosine-5-O-diphosphate (2MeSADP), induced similar effects to ATP with an EC(50) of 160 +/- 30 nm. The effects of 250 nm2MeSADP were blocked by 500 nmMRS2179, a specific P2Y(1) receptor inhibitor, suggesting that ATP acts on P2Y(1) receptors to inhibit chloride channels. The inhibition of chloride channels by extracellular ATP has implications for muscle excitability and fatigue, and the pathophysiology of myotonias. PMID- 19805743 TI - The response to paired motor cortical stimuli is abolished at a spinal level during human muscle fatigue. AB - During maximal exercise, supraspinal fatigue contributes significantly to the decline in muscle performance but little is known about intracortical inhibition during such contractions. Long-interval inhibition is produced by a conditioning motor cortical stimulus delivered via transcranial magnetic stimulation (TMS) 50 200 ms prior to a second test stimulus. We aimed to delineate changes in this inhibition during a sustained maximal voluntary contraction (MVC). Eight subjects performed a 2 min MVC of elbow flexors. Single test and paired (conditioning-test interval of 100 ms) stimuli were delivered via TMS over the motor cortex every 7 8 s throughout the effort and during intermittent MVCs in the recovery period. To determine the role of spinal mechanisms, the protocol was repeated but the TMS test stimulus was replaced by cervicomedullary stimulation which activates the corticospinal tract. TMS motor evoked potentials (MEPs) and cervicomedullary motor evoked potentials (CMEPs) were recorded from biceps brachii. Unconditioned MEPs increased progressively with fatigue, whereas CMEPs increased initially but returned to the control value in the final 40 s of contraction. In contrast, both conditioned MEPs and CMEPs decreased rapidly with fatigue and were virtually abolished within 30 s. In recovery, unconditioned responses required <30 s but conditioned MEPs and CMEPs required 90 s to return to control levels. Thus, long interval inhibition increased markedly as fatigue progressed. Contrary to expectations, subcortically evoked CMEPs were inhibited as much as MEPs. This new phenomenon was also observed in the first dorsal interosseous muscle. Tested with a high intensity conditioning stimulus during a fatiguing maximal effort, long interval inhibition of MEPs was increased primarily by spinal rather than motor cortical mechanisms. The spinal mechanisms exposed here may contribute to the development of central fatigue in human muscles. PMID- 19805744 TI - PKC-permitted elevation of sarcolemmal KATP concentration may explain female specific resistance to myocardial infarction. AB - The female myocardium, relative to that of the male, exhibits sustained resistance to ischaemic tissue injury, a phenomenon termed sex-specific cardioprotection (SSC). SSC is dependent upon the sarcolemmal K(ATP) channel (sarcK(ATP)), and protein kinase C (PKC). Here we investigate whether PKC mediated regulation of sarcK(ATP) concentration can explain this endogenous form of protection. Hearts from male (M) and female (F) rats were Langendorff-perfused for 30 min prior to either regional ischaemia-reperfusion (I/R), or global ischaemia (GISC). For both protocols, pre-ischaemic blockade of PKC was achieved by chelerythrine (Chel) in male (M + C) and female (F + C) hearts. Additional female hearts underwent sarcK(ATP) antagonism during I/R by HMR-1098 (HMR), either alone or in combination with Chel (HMR + Chel). GISC hearts were fractionated to assess cellular distribution of PKC and sarcK(ATP). Sex-specific infarct resistance was apparent under control I/R (F, 23 +/- 3% vs. M, 36 +/- 4%, P < 0.05) and abolished by Chel (F + C, 36 +/- 3%). Female infarct resistance was susceptible to sarcK(ATP) blockade (Control, 16 +/- 2% vs. HMR, 27 +/- 3%), and PKC blockade had no additional effect (HMR + Chel, 26 +/- 2%). The prevalence of Kir6.2 and SUR2 was higher in the sarcolemmal fractions of females (Kir6.2: F, 1.24 +/- 0.07 vs. M, 1.02 +/- 0.06; SUR2: F, 3.16 +/- 0.22 vs. M, 2.45 +/- 0.09; ratio units), but normalized by Chel (Kir6.2: F, 1.06 +/- 0.07 vs. M, 0.99 +/- 0.06; SUR2: F, 2.99 +/- 0.09 vs. M, 2.82 +/- 0.22, M; ratio units). Phosphorylation of sarcolemmal PKC was reduced by Chel (p-PKC/PKC: control, 0.43 +/- 0.02; Chel, 0.29 +/- 0.01; P < 0.01). We conclude that PKC-mediated regulation of sarcK(ATP) may account for the physiologically sustainable dependence of SSC upon both PKC and sarcK(ATP), and that this regulation involves PKC-permitted enrichment of the female sarcolemma with sarcK(ATP). As such, the PKC-sarcK(ATP) axis may represent a target for sustainable prophylactic induction of cardioprotection. PMID- 19805745 TI - Episodic spinal serotonin receptor activation elicits long-lasting phrenic motor facilitation by an NADPH oxidase-dependent mechanism. AB - Phrenic long-term facilitation (pLTF) is a serotonin (5-HT)-dependent augmentation of phrenic motor output induced by acute intermittent hypoxia (AIH). AIH-induced pLTF requires spinal NADPH oxidase activity and reactive oxygen species (ROS) formation. Since 5-HT receptor activation stimulates NADPH oxidase activity in some cell types, we tested the hypothesis that episodic spinal 5-HT receptor activation (without AIH) is sufficient to elicit an NADPH oxidase dependent facilitation of phrenic motor output (pMF). In anaesthetised, artificially ventilated adult male rats, episodic intrathecal 5-HT injections (3 x 6 microl injections at 5 min intervals) into the cerebrospinal fluid (CSF) near cervical spinal segments containing the phrenic motor nucleus elicited a progressive increase in integrated phrenic nerve burst amplitude (i.e. pMF) lasting at least 60 min post-5-HT administration. Hypoglossal (XII) nerve activity was unaffected, suggesting that effective doses of 5-HT did not reach the brainstem. A single 5-HT injection was without effect. 5-HT-induced pMF was dose dependent, but exhibited a bell-shaped dose-response curve. Activation of different 5-HT receptor subtypes, specifically 5-HT(2) versus 5-HT(7) receptors, may underlie the bell-shaped dose-response curve via a mechanism of 'cross-talk' inhibition. Pre-treatment with NADPH oxidase inhibitors, apocynin or diphenylenodium (DPI), blocked 5-HT induced pMF. Thus, episodic spinal 5-HT receptor activation is sufficient to elicit pMF by an NADPH oxidase-dependent mechanism, suggesting common mechanisms of ROS formation with AIH-induced pLTF. An understanding of the mechanisms giving rise to AIH-induced pLTF and 5-HT induced pMF may inspire novel therapeutic strategies for respiratory insufficiency in diverse conditions, such as sleep apnoea, cervical spinal injury or amyotrophic lateral sclerosis. PMID- 19805746 TI - Impact of transcranial direct current stimulation on spinal network excitability in humans. AB - Transcranial direct current stimulation (tDCS) when applied over the motor cortex, modulates excitability dependent on the current polarity. The impact of this cortical modulation on spinal cord network excitability has rarely been studied. In this series of experiments, performed in healthy subjects, we show that anodal tDCS increases disynaptic inhibition directed from extensor carpi radialis (ECR) to flexor carpi radialis (FCR) with no modification of presynaptic inhibition of FCR Ia terminals and FCR H-reflex recruitment curves. We also show that cathodal tDCS does not modify spinal network excitability. Our results suggest that the increase of disynaptic inhibition observed during anodal tDCS relies on an increase of disynaptic interneuron excitability and that tDCS over the motor cortex in human subjects induces effects on spinal network excitability. Our results highlight the fact that the effects of tDCS should be considered in regard to spinal motor circuits and not only to cortical circuits. PMID- 19805747 TI - Progressive augmentation and ventilatory long-term facilitation are enhanced in sleep apnoea patients and are mitigated by antioxidant administration. AB - Progressive augmentation (PA) and ventilatory long-term facilitation (vLTF) of respiratory motor output are forms of respiratory plasticity that are initiated during exposure to intermittent hypoxia. The present study was designed to determine whether PA and vLTF are enhanced in obstructive sleep apnoea (OSA) participants compared to matched healthy controls. The study was also designed to determine whether administration of an antioxidant cocktail mitigates PA and vLTF. Thirteen participants with sleep apnoea and 13 controls completed two trials. During both trials participants were exposed to intermittent hypoxia which included twelve 4-min episodes of hypoxia (P(ETCO(2)), 50 mmHg; P(ETCO(2)), 4 mmHg above baseline) followed by 30 min of recovery. Prior to exposure to intermittent hypoxia, participants were administered, in a randomized fashion, either an antioxidant or a placebo cocktail. Baseline measures of minute ventilation during the placebo and antioxidant trials were not different between or within groups. During the placebo trial, PA was evident in both groups; however it was enhanced in the OSA group compared to control (last hypoxic episode 36.9 +/- 2.8 vs. 27.7 +/- 2.2 l min(-1); P or=0.82 mm); likelihood ratio 2.2 (1.9-2.5). The results were similar in case control and cohort studies. CONCLUSION: Neither carotid plaque nor IMT has a CHD screening performance that is sufficiently discriminatory between affected and unaffected individuals to be a worthwhile screening test. PMID- 19805757 TI - Combining carotid intima-media thickness with carotid plaque on screening for coronary heart disease. AB - BACKGROUND: Ultrasound-detected carotid artery intima-media thickness (IMT) and carotid plaque are possible screening tests for coronary heart disease (CHD) among asymptomatic individuals. OBJECTIVE: To assess the increase in screening performance of combining carotid IMT and plaque compared with each measurement alone in the identification of individuals with CHD. METHODS: Ultrasound examination of left and right carotid arteries was performed on 100 individuals (median age 57), 55 with a history of CHD (unstable angina or myocardial infarction) and 45 without. IMT measurements were taken from the common carotid artery and plaque was identified above, at and below the carotid bifurcation. Associations between IMT and plaque were determined using logistic regression, and screening performance was assessed from the distributions of IMT and plaque among cases and controls. RESULTS: At a false-positive rate of 5%, IMT (cut-off >0.75 mm) identified 30% (95% CI 14-58) of affected individuals. There was an increase in the detection rate of 8 percentage points (1-33%) using IMT and plaque combined compared with IMT alone. As the false-positive increased, the difference in the detection rate increased, up to a maximum of 20 percentage points (5-38%) at a false-positive rate of 20%. The comparison of IMT and plaque combined with plaque alone could only be estimated for the false-positive rate observed using plaque alone (18%); at this point the detection rate was 72% for plaque and 75% for plaque and IMT combined, an increase of 3 percentage points (0 4%). CONCLUSION: In screening for CHD, combining carotid IMT measurement with plaque assessment is better than using either measurement alone, but the improvement in discrimination is not sufficient to make carotid ultrasound screening for CHD worthwhile. PMID- 19805764 TI - Summaries for patients. The effects of hand washing and facemasks on prevention of influenza infection. PMID- 19805765 TI - Summaries for patients. The effects of limited sleep and alcohol on driving performance in people with untreated sleep apnea. PMID- 19805766 TI - Summaries for patients. Using nontraditional risk factors to estimate risk for coronary heart disease. PMID- 19805767 TI - In the clinic. Community-acquired pneumonia. PMID- 19805768 TI - Effects of alcohol and sleep restriction on simulated driving performance in untreated patients with obstructive sleep apnea. AB - BACKGROUND: Because of previous sleep disturbance and sleep hypoxia, patients with obstructive sleep apnea (OSA) might be more vulnerable to the effects of alcohol and sleep restriction than healthy persons. OBJECTIVE: To compare the effects of sleep restriction and alcohol on driving simulator performance in patients with OSA and age-matched control participants. DESIGN: Driving simulator assessments in 2 groups under 3 different conditions presented in random order. SETTING: Adelaide Institute for Sleep Health, Sleep Laboratory, Adelaide, Australia. PARTICIPANTS: 38 untreated patients with OSA and 20 control participants. MEASUREMENTS: Steering deviation, crashes, and braking reaction time. INTERVENTION: Unrestricted sleep, sleep restricted to a maximum of 4 hours, and ingestion of an amount of 40% vodka calculated to achieve a blood alcohol level of 0.05 g/dL. RESULTS: Patients with OSA demonstrated increased steering deviation compared with control participants (mean, 50.5 cm [95% CI, 46.1 to 54.9 cm] in the OSA group and 38.4 cm [CI, 32.4 to 44.4 cm] in the control group; P < 0.01) and significantly greater steering deterioration over time (group by time interaction, P = 0.02). The increase in steering deviation after sleep restriction and alcohol was approximately 40% greater in patients with OSA than in control participants (group by condition interaction, P = 0.04). Patients with OSA crashed more frequently than control participants (1 vs. 24 participants; odds ratio [OR], 25.4; P = 0.03) and crashed more frequently after sleep restriction (OR, 4.0; P < 0.01) and alcohol consumption (OR, 2.3; P = 0.02) than after normal sleep. In patients with OSA, prolonged eye closure (>2 seconds) and microsleeps (> 2 seconds of theta activity on electroencephalography) were significant crash predictors (OR, 19.2 and 7.2, respectively; P < 0.01). Braking reaction time was slower after sleep restriction than after normal sleep (mean, 1.39 [SD, 0.06] seconds vs. 1.22 [SD, 0.04] seconds; P < 0.01) but not after alcohol consumption. No group differences were found. LIMITATION: Simulated driving was assessed rather than on-road driving. CONCLUSION: Patients with OSA are more vulnerable than healthy persons to the effects of alcohol consumption and sleep restriction on various driving performance variables. PRIMARY FUNDING SOURCE: Australian National Health and Medical Research Council. PMID- 19805769 TI - Associations between structural capabilities of primary care practices and performance on selected quality measures. AB - BACKGROUND: Recent proposals to reform primary care have encouraged physician practices to adopt such structural capabilities as performance feedback and electronic health records. Whether practices with these capabilities have higher performance on measures of primary care quality is unknown. OBJECTIVE: To measure associations between structural capabilities of primary care practices and performance on commonly used quality measures. DESIGN: Cross-sectional analysis. SETTING: Massachusetts. PARTICIPANTS: 412 primary care practices. MEASUREMENTS: During 2007, 1 physician from each participating primary care practice (median size, 4 physicians) was surveyed about structural capabilities of the practice (responses representing 308 practices were obtained). Data on practice structural capabilities were linked to multipayer performance data on 13 Healthcare Effectiveness Data and Information Set (HEDIS) process measures in 4 clinical areas: screening, diabetes, depression, and overuse. RESULTS: Frequently used multifunctional electronic health records were associated with higher performance on 5 HEDIS measures (3 in screening and 2 in diabetes), with statistically significant differences in performance ranging from 3.1 to 7.6 percentage points. Frequent meetings to discuss quality were associated with higher performance on 3 measures of diabetes care (differences ranging from 2.3 to 3.1 percentage points). Physician awareness of patient experience ratings was associated with higher performance on screening for breast cancer and cervical cancer (1.9 and 2.2 percentage points, respectively). No other structural capabilities were associated with performance on more than 1 measure. No capabilities were associated with performance on depression care or overuse. LIMITATION: Structural capabilities of primary care practices were assessed by physician survey. CONCLUSION: Among the investigated structural capabilities of primary care practices, electronic health records were associated with higher performance across multiple HEDIS measures. Overall, the modest magnitude and limited number of associations between structural capabilities and clinical performance suggest the importance of continuing to measure the processes and outcomes of care for patients. PRIMARY FUNDING SOURCE: The Commonwealth Fund. PMID- 19805770 TI - Using nontraditional risk factors in coronary heart disease risk assessment: U.S. Preventive Services Task Force recommendation statement. AB - DESCRIPTION: New recommendation from the U.S. Preventive Services Task Force (USPSTF) on the use of nontraditional, or novel, risk factors in assessing the coronary heart disease (CHD) risk of asymptomatic persons. METHODS: Systematic reviews were conducted of literature since 1996 on 9 proposed nontraditional markers of CHD risk: high-sensitivity C-reactive protein, ankle-brachial index, leukocyte count, fasting blood glucose, periodontal disease, carotid intima-media thickness, coronary artery calcification score on electron-beam computed tomography, homocysteine, and lipoprotein(a). The reviews followed a hierarchical approach aimed at determining which factors could practically and definitively reassign persons assessed as intermediate-risk according to their Framingham score to either a high-risk or low-risk strata, and thereby improve outcomes by means of aggressive risk-factor modification in those newly assigned to the high risk stratum. RECOMMENDATION: The USPSTF concludes that the current evidence is insufficient to assess the balance of benefits and harms of using the nontraditional risk factors studied to screen asymptomatic men and women with no history of CHD to prevent CHD events. (I statement). PMID- 19805771 TI - C-reactive protein as a risk factor for coronary heart disease: a systematic review and meta-analyses for the U.S. Preventive Services Task Force. AB - BACKGROUND: C-reactive protein (CRP) may help to refine global risk assessment for coronary heart disease (CHD), particularly among persons who are at intermediate risk on the basis of traditional risk factors alone. PURPOSE: To assist the U.S. Preventive Services Task Force (USPSTF) in determining whether CRP should be incorporated into guidelines for CHD risk assessment. DATA SOURCES: MEDLINE search of English-language articles (1966 to November 2007), supplemented by reference lists of reviews, pertinent studies, editorials, and Web sites and by expert suggestions. STUDY SELECTION: Prospective cohort, case-cohort, and nested case-control studies relevant to the independent predictive ability of CRP when used in intermediate-risk persons. DATA EXTRACTION: Included studies were reviewed according to predefined criteria, and the quality of each study was rated. DATA SYNTHESIS: The validity of the body of evidence and the net benefit or harm of using CRP for CHD risk assessment were evaluated. The combined magnitude of effect was determined by meta-analysis. The body of evidence is of good quality, consistency, and applicability. For good studies that adjusted for all Framingham risk variables, the summary estimate of relative risk for incident CHD was 1.58 (95% CI, 1.37 to 1.83) for CRP levels greater than 3.0 mg/L compared with levels less than 1.0 mg/L. Analyses from 4 large cohorts were consistent in finding evidence that including CRP improves risk stratification among initially intermediate-risk persons. C-reactive protein has desirable test characteristics, and good data exist on the prevalence of elevated CRP levels in intermediate-risk persons. Limited evidence links changes in CRP level to primary prevention of CHD events. LIMITATIONS: Study methods for measuring Framingham risk variables and other covariates varied. Ethnic and racial minority populations were poorly represented in most studies, limiting generalizability. Few studies directly assessed the effect of CRP on risk reclassification in intermediate-risk persons. CONCLUSION: Strong evidence indicates that CRP is associated with CHD events. Moderate, consistent evidence suggests that adding CRP to risk prediction models among initially intermediate-risk persons improves risk stratification. However, sufficient evidence that reducing CRP levels prevents CHD events is lacking. PMID- 19805772 TI - Emerging risk factors for coronary heart disease: a summary of systematic reviews conducted for the U.S. Preventive Services Task Force. AB - BACKGROUND: Traditional risk factors do not explain all of the risk for incident coronary heart disease (CHD) events. Various new or emerging risk factors have the potential to improve global risk assessment for CHD. PURPOSE: To summarize the results of 9 systematic reviews of novel risk factors to help the U.S. Preventive Services Task Force (USPSTF) evaluate the factors' clinical usefulness. DATA SOURCES: Results from a MEDLINE search for English-language articles published from 1966 to September 2008, using the Medical Subject Heading terms cohort studies and cardiovascular diseases in combination with terms for each risk factor. STUDY SELECTION: Studies were included if the participants had no baseline cardiovascular disease and the investigators adjusted for at least 6 Framingham risk factors. DATA EXTRACTION: Study quality was evaluated by using USPSTF criteria and overall quality of evidence for each risk factor by using a modified version of the Grading of Recommendations, Assessment, Development, and Evaluation framework. Each factor's potential clinical value was evaluated by using a set of criteria that emphasized the importance of the effect of that factor on the reclassification of intermediate-risk persons. DATA SYNTHESIS: 9 systematic reviews were conducted. C-reactive protein (CRP) was the best candidate for use in screening and the most rigorously studied, but evidence that changes in CRP level lead to primary prevention of CHD events is inconclusive. The other evaluated risk factors were coronary artery calcium score as measured by electron-beam computed tomography, lipoprotein(a) level, homocysteine level, leukocyte count, fasting blood glucose, periodontal disease, ankle-brachial index, and carotid intima-media thickness. The availability and validity of the evidence varied considerably across the risk factors in terms of aggregate quality, consistency of findings, and applicability to intermediate-risk persons in the general population. For most risk factors, no studies assessed their usefulness for reclassifying intermediate-risk persons. LIMITATIONS: Because of lack of access to original data, no firm conclusions could be drawn about differences in risk prediction among racial and ethnic groups. The review did not emphasize within-cohort comparisons of multiple risk factors. CONCLUSION: The current evidence does not support the routine use of any of the 9 risk factors for further risk stratification of intermediate-risk persons. PMID- 19805775 TI - Redundant data in the meta-analysis on Helicobacter pylori eradication. PMID- 19805776 TI - Granulysin as a marker for early diagnosis of the Stevens-Johnson syndrome. PMID- 19805777 TI - Localized amyloidosis at the site of enfuvirtide injection. PMID- 19805778 TI - Creation of a scholars program in dental leadership (SPDL) for dental and dental hygiene students. AB - There is a great need for leaders in the dental profession. As technological advances make our world smaller and our lives faster and more complex, we as a profession face challenges and opportunities that are evolving. Many of the changes in the scope and mode of practice will require new and different approaches. Meeting these challenges will require changes in how we as dental professionals do business; interact with our patients, other stakeholders, and health care providers; and educate our future colleagues. The purposeful incorporation of leadership education into dental and dental hygiene curricula represents an important departure from existing paradigms-but will help prepare our students to address these challenges. This article provides an overview of the development of a Scholars Program in Dental Leadership (SPDL) at the University of Michigan School of Dentistry. Our aim for the program is to create a learning environment that fosters leadership development, so that students are prepared and motivated to assume leadership positions in the profession and their communities. PMID- 19805779 TI - Using competencies to improve dental practice management education. AB - The ADEA Competencies for the New General Dentist contain a significant number of practice management-related competencies. To date, these have been taught primarily in a lecture format in the third and fourth years of the dental curriculum. Presenting information in this way only satisfies the lower level learning skills, not the skills needed to become a competent general dentist. This article presents a framework for applying the competency-based education model to dental practice management teaching, learning, and assessment. PMID- 19805780 TI - Full-time dental faculty perceptions of satisfaction with the academic work environment. AB - A significant factor in a faculty member's accepting or maintaining an academic appointment is the work environment. Assessing the work environment to identify characteristics that could increase faculty retention and recruitment could be valuable to an educational institution. This study assessed the academic dental work environment to identify positive and negative areas affecting career satisfaction. An online survey about departmental structure and individual work patterns was sent to the deans of fifty-two U.S. dental schools who then forwarded the survey to their faculty. Thirty-eight institutions (73 percent) and 451 full-time faculty members from those thirty-eight schools responded. Most dental faculty members in this survey intend to remain in academia for the next five to eight years. Slightly fewer male faculty members intend to remain in dental education for five to eight years than do female faculty members. Positive satisfaction aspects of the work environment listed by respondents included supportive chair/administration, working relationships with colleagues, and interactions with students. Negative satisfaction aspects of the work environment included low salary, long hours, and heavy workloads. Both positive aspects of job satisfaction and negative factors that impede productivity need to be analyzed within the framework of each institution to enact change for career enrichment, leading to increased faculty recruitment and retention. PMID- 19805781 TI - Introducing infant oral health into dental curricula: a clinical intervention. AB - Although pediatric dentists are trained to address the oral health needs of young children, few general practitioners receive this training in dental school. The purpose of this study was to evaluate change in dental students' knowledge, confidence, opinions, and behaviors following a curricular intervention in infant and toddler oral health. Using a pre- and post-survey study design, forty-five intervention and forty-one control group students participated in the study. The intervention consisted of a three-hour seminar, followed by three or four clinical sessions. Descriptive statistics and analysis of covariance were used to compare the average scores of the two groups after adjusting for pre-construct scores. The response rate was 84 percent. The mean scores of the control and intervention groups did not differ significantly for any of the constructs at baseline (p>.14). Bivariate analysis showed the average post-scores differed significantly (p<.01) in the control and intervention groups for all constructs after adjusting for pre-scores. Similar findings occurred in the regression modeling, with previous experience caring for young children, ability to speak Spanish, and gender influencing the outcomes. Eighty-eight percent of the students enrolled in the intervention stated they were more likely to treat children in this age group following this clinical experience. PMID- 19805782 TI - Intelligent tutoring system for clinical reasoning skill acquisition in dental students. AB - Learning clinical reasoning is an important core activity of the modern dental curriculum. This article describes an intelligent tutoring system (ITS) for clinical reasoning skill acquisition. The system is designed to provide an experience that emulates that of live human-tutored problem-based learning (PBL) sessions as much as possible, while at the same time permitting the students to participate collaboratively from disparate locations. The system uses Bayesian networks to model individual student knowledge and activity, as well as that of the group. Tutoring algorithms use the models to generate tutoring hints. The system incorporates a multimodal interface that integrates text and graphics so as to provide a rich communication channel between the students and the system, as well as among students in the group. Comparison of learning outcomes shows that student clinical reasoning gains from the ITS are similar to those obtained from human-tutored sessions. PMID- 19805783 TI - The role of basic sciences in diagnostic oral radiology. AB - Although it is generally taken for granted that dental education must include both basic science and feature-based knowledge components, little is known about their relative roles in visual interpretation of radiographs. The objectives of this study were twofold. First, we sought to compare the educational efficacy of three learning strategies in diagnostic radiology: one that used basic scientific (pathophysiologic) information, one that used feature lists structured with an organizational tool, and one that used unstructured feature lists. Our second objective was to determine whether basic scientific information provides conceptual coherence or is merely a simple means for organizing feature-based knowledge. Predoctoral dental and undergraduate dental hygiene students (n=96) were randomly assigned into three groups (basic science, structured algorithm, and feature list) and were taught four confusable intrabony entities. The students completed diagnostic and memory tests immediately after learning and one week later, and these data were subjected to a 3x2 repeated measures ANOVA. For the diagnostic test, students in the basic science group outperformed those assigned to the feature list and structured algorithm groups on immediate and delayed testing (p<0.05). A main effect of learning condition was found to be significant. On the memory test, performance was similar across all three groups, and no significant effects were found. The results of this study support the critical role of basic scientific knowledge in diagnostic radiology. This study also refutes the organized learning theory and provides support for the conceptual coherence theory as a possible explanation for the process by which basic science aids in diagnosis. PMID- 19805784 TI - A clinically oriented complete denture program for second-year dental students. AB - The traditional preclinical complete denture prosthodontic curriculum relies predominantly on the laboratory (e.g., bench-type) component of the complete denture fabrication process. In most cases, this involves a passive model of student knowledge acquisition utilizing lectures and low-fidelity laboratory exercises. A recently implemented program in the College of Dentistry at the University of Illinois at Chicago challenges this educational paradigm by introducing an active learning environment for second-year students based on significant clinical exposure with patients. The result is a major shift of emphasis, first, from a purely technical/laboratory aspect of the discipline to patient-centered education and, second, from mastering individual phases of denture fabrication to understanding the entire process of edentulous patient care. To compare student outcomes in the new program with those in the traditional program, their performance overall and in three components of the final examination for each program were statistically compared. The results of the one-way ANOVA analysis show statistically significant improvement in the students' total score in the new program, including their performance on the written, practical, and OSCE portions of the final examination. This article describes the rationale, logistics, challenges, and advantages of the new educational model of the complete denture prosthodontics curriculum. PMID- 19805785 TI - Implementing e-learning in a radiological science course in dental education: a short-term longitudinal study. AB - This short-term longitudinal case study evaluated the quality of learning by comparing assessment outcomes following the traditional face-to-face lecture teaching of a radiological science course to first-year dental students in 2007 with assessment outcomes following conversion of the course to an e-learning format for first-year dental students in 2008. Multiple methods were used to triangulate and add rigor to the study. Not only were multiple-choice exam results from both the 2007 and 2008 cohorts statistically compared, but an interview was conducted with the content provider to capture a different perspective. A survey was also used to provide insights into students' perceptions of the use of e-learning. Finally, the tracking records of students who accessed online lectures were considered in analysis of the data. Due to the positive response from the students, coupled with the nature of the course and an emphasis on the quality of the content, e-learning could be fully implemented, despite some shortcomings, without compromising the quality of learning. PMID- 19805786 TI - Transition of a dental histology course from light to virtual microscopy. AB - The transition of the dental histology course at the University of Texas Health Science Center at San Antonio Dental School was completed gradually over a five year period. A pilot project was initially conducted to study the feasibility of integrating virtual microscopy into a traditional light microscopic lecture and laboratory course. Because of the difficulty of procuring quality calcified and decalcified sections of teeth, slides from the student loan collection in the oral histology block of the course were outsourced for conversion to digital images and placed on DVDs along with a slide viewer. The slide viewer mimicked the light microscope, allowing horizontal and vertical movement and changing of magnification, and, in addition, a feature to capture static images. In a survey, students rated the ease of use of the software, quality of the images, maneuverability of the images, and questions regarding use of the software, effective use of laboratory, and faculty time. Because of the positive support from the students, our entire student loan collection of 153 glass slides was subsequently converted to virtual images and distributed on an Apricorn pocket external hard drive. Students were asked to assess the virtual microscope over a four-year period. As a result of the surveys, light microscopes have been totally eliminated, and microscope exams have been replaced with project slide examinations. In the future, we plan to expand our virtual slides and incorporate computer testing. PMID- 19805787 TI - Practicing dental hygienists' perceptions about the bachelor of science in dental hygiene and the oral health practitioner. AB - No research data are available about practicing dental hygienists' opinions regarding the Bachelor of Science in Dental Hygiene (B.S.D.H.) and the oral health practitioner (OHP), a new professional category in dental hygiene; however, such views would be useful to those implementing these programs in the academic setting as well as those involved in proposing and passing legislation regarding them. The purpose of our study was to gather information from a group of practicing dental hygienists regarding their opinions in three areas: 1) the entry-level B.S.D.H., 2) the OHP, and 3) reasons for being for or against these programs. A survey, sent to 564 dental hygiene graduates, used a five-point Likert scale to evaluate perceptions in various categories. The respondents also ranked perceived benefits and negative impacts. The usable return rate was 33.6 percent. Descriptive statistics were developed, and chi-square tests were used to analyze the data. More than 70 percent of the respondents agreed that an associate's degree sufficiently prepared dental hygienists for their positions and that the OHP would have a positive impact on access to dental care. The majority also said they felt the master's-educated hygienist would be adequately prepared to perform proposed OHP functions. Selected demographic variables were found to be significantly associated with perceptions, including that the B.S.D.H. was viewed more positively by younger respondents. PMID- 19805788 TI - In Memoriam: Sabra F. Woolley, PhD. PMID- 19805789 TI - Behavioral theory in a diverse society: like a compass on Mars. AB - The behavioral theory constructs most often used to study mammography utilization perceived benefit, perceived susceptibility, self-efficacy, intention, and subjective norms-have neither been developed nor sufficiently tested among diverse racial/ethnic subgroups. The authors explored these constructs and their underlying assumptions relating to the social context of Filipina and Latina women. The mixed-methods study included testing construct measures in the multilingual surveys of a concurrent intervention study of 1,463 women from five ethnic groups. An intensive inductive investigation then targeted Latina and Filipina women to elucidate connections between social context and individual screening behavior. In-depth interviews were conducted with 11 key informant scholars, 13 community gatekeepers, and 29 lay women, and a supplemental study videotaped and interviewed 9 mother-daughter dyads. Three social context domains emerged: relational culture, social capital, and transculturation and transmigration. The meaning and appropriateness of the five behavioral constructs were analyzed in relation to these domains. In contradistinction to tenets of behavioral theory, the authors found that social context can influence behavior directly, circumventing or attenuating the influence of individual beliefs; contextual influences, synthesized from multiple perspectives, can operate at an unconscious level not accessible to the individual; and contextual influences are dynamic, contingent on distal and proximal forces coming together in a given moment and are thus not consistent with an exclusive focus at the individual level. This article describes the study methods, summarizes main findings, and preview the detailed results presented in the other articles in this issue. PMID- 19805790 TI - Behavioral constructs and mammography in five ethnic groups. AB - Intention, self-efficacy, perceived susceptibility, perceived benefits, and subjective norms are key constructs of health behavior theories; their predictive validity for cancer screening has not been ascertained in multiethnic populations. Participants were 1,463 African American, Chinese, Filipina, Latina, and White women aged 40 to 74 interviewed by telephone in their preferred languages. The relationship between baseline constructs and mammography 2 years later was assessed using multivariable logistic regression. Intention predicted mammography overall and among Whites (odds ratio [OR] = 5.0, 95% confidence interval [CI] = 2.4, 10), with racial/ethnic differences in association (p = .020). Self-efficacy predicted mammography overall and among Whites (OR = 3.5, 95% CI = 1.1, 11), with no racial/ethnic interaction. Perceived benefits and subjective norms were associated with screening overall and in some racial/ethnic groups. These results generally support cross-cultural applicability of four of the five constructs to screening with mixed predictive value of measures across racial/ethnic groups. Additional in-depth inquiry is required to refine assessment of constructs. PMID- 19805791 TI - Theorizing social context: rethinking behavioral theory. AB - Major behavioral theories focus on proximal influences on behavior that are considered to be predominantly cognitive characteristics of the individual largely uninfluenced by social context. Social ecological models integrate multiple levels of influence on health behavior and are noted for emphasizing the interdependence of environmental settings and life domains. This theory-based article explains how social context is conceptualized in the social sciences and how the social science conceptualization differs from and can broaden the analytic approach to health behavior. The authors use qualitative data from the Behavioral Constructs and Culture in Cancer Screening study to illustrate our conceptualization of social context. We conclude that the incorporation into health behavior theory of a multidimensional socioculturally oriented, theoretical approach to social context is critical to understand and redress health disparities in multicultural societies like the United States. PMID- 19805792 TI - Perceived susceptibility to illness and perceived benefits of preventive care: an exploration of behavioral theory constructs in a transcultural context. AB - This article describes how the social context of transculturation (cultural change processes) and transmigration (migration in which relationships are sustained across national boundaries) can directly influence use of mammography screening. The authors conducted semistructured interviews with Latino and Filipino academics and social service providers and with U.S.-born and immigrant Latinas and Filipinas to explore direct and indirect influences of social context on health behavior (Behavioral Constructs and Culture in Cancer Screening study). Iterative analyses identified themes of the transcultural domain: colonialism, immigration, discrimination, and therapeutic engagement. In this domain, the authors examine two key behavioral theory constructs, perceptions of susceptibility to illness and perceptions of benefits of preventive medical care. The findings raise concerns about interventions to promote mammography screening primarily based on provision of scientific information. The authors conclude that social context affects behavior directly rather than exclusively through beliefs as behavioral theory implies and that understanding contextual influences, such as transculturation, points to different forms of intervention. PMID- 19805793 TI - Intention, subjective norms, and cancer screening in the context of relational culture. AB - Research targeting disparities in breast cancer detection has mainly utilized theories that do not account for social context and culture. Most mammography promotion studies have used a conceptual framework centered in the cognitive constructs of intention (commonly regarded as the most important determinant of screening behavior), self-efficacy, perceived benefit, perceived susceptibility, and/or subjective norms. The meaning and applicability of these constructs in diverse communities are unknown. The purpose of this study is to inductively explore the social context of Filipina and Latina women (the sociocultural forces that shape people's day-to-day experiences and that directly and indirectly affect health and behavior) to better understand mammography screening behavior. One powerful aspect of social context that emerged from the findings was relational culture, the processes of interdependence and interconnectedness among individuals and groups and the prioritization of these connections above virtually all else. The authors examine the appropriateness of subjective norms and intentions in the context of relational culture and identify inconsistencies that suggest varied meanings from those intended by behavioral theorists. PMID- 19805794 TI - Social and cultural meanings of self-efficacy. AB - This article describes the influences of social context on women's health behavior through illustration of the powerful influences of social capital (the benefits and challenges that accrue from participation in social networks and groups) on experiences and perceptions of self-efficacy. The authors conducted inductive interviews with Latino and Filipino academics and social service providers and with U.S.-born and immigrant Latinas and Filipinas to explore direct and indirect influences of social context on health behaviors such as mammography screening. Iterative thematic analysis identified themes (meanings of efficacy, spheres of efficacy, constraints on efficacy, sources of social capital, and differential access to and quality of social capital) that link the domain of social capital with the behavioral construct perceived self-efficacy. The authors conclude that social capital addresses aspects of social context absent in the current self-efficacy construct and that these aspects have important implications for scholars' and practitioners' understandings of health behavior and intervention development. PMID- 19805795 TI - Adult daughters' influence on mothers' health-related decision making: an expansion of the subjective norms construct. AB - This study of mother-adult daughter communication uses qualitative methods to explore the appropriateness of including adult daughters as referents in the measurement of subjective norms (a behavioral theory construct) related to the use of mammography and other health-related tests and services. The methods were chosen to approximate as closely as possible the mother-adult daughter relationship in the context of daily life. This inductive approach contrasts with the deductive origins of the construct. A sample of nine Mexican and Filipina immigrant and U.S.-born mothers and their adult daughters was recruited. Data were collected in two phases: (a) videotaped observations of mother-daughter dyads discussing health-related topics and (b) follow-up interviews designed to obtain an emic (insider) perspective of the videotaped interaction. Results show that adult daughters influence their mothers' ability to navigate the health care system and contribute to health-related decision making and behavior, suggesting that it may be appropriate to include adult daughters in the assessment of subjective norms. PMID- 19805796 TI - Triple therapy for chronic obstructive pulmonary disease: trials catching up with clinical practice? PMID- 19805797 TI - Prediction of perioperative mortality after lung cancer resection in the elderly. PMID- 19805798 TI - Effect of vitamin D on tuberculosis and HIV replication depends on conversion to calcitriol and concentration. PMID- 19805799 TI - Adolf Kussmaul: distinguished clinician and medical pioneer. PMID- 19805800 TI - The case of the proliferating paradigms. PMID- 19805801 TI - Patient real-time and 12-month retrospective perceptions of difficult communications in the cancer diagnostic period. AB - Communication is a notoriously complex challenge in the cancer care context. Our program of research involves exploration of patient-provider communications across the cancer trajectory from the patient perspective.Toward this end, we have been following a cohort of 60 cancer patients, representing a range of tumor sites, from immediately after diagnosis through to recovery, chronic, or advanced disease. Drawing on interpretive description analytic techniques, we documented patterns and themes related to various components of the cancer journey. In this article, we report on findings pertaining to poor communication during the initial diagnostic period, as described by patients at the time of diagnosis and 1 year later.These findings illuminate the dynamics of communication problems during that complex period, and depict the mechanisms by which patients sought to confront these challenges to optimize their cancer care experience. On the basis of these findings, considered in the context of the body of available evidence, suggestions are proposed as to appropriate directions for system-level solutions to the complex communication challenges within cancer care. PMID- 19805802 TI - Accounts of HIV seroconversion among substance-using gay and bisexual men. AB - Statistical associations between substance use and seroconversion among gay and bisexual men abound. However, these associations often ignore men's own interpretations of their seroconversion. Using in-depth interviews with gay and bisexual men who reported using drugs or alcohol at the time of their seroconversion, we identify how these men explain the events that led to HIV transmission. Whereas a small minority of respondents reported substance use to explain their seroconversion, the majority reported three competing explanations. These participants claimed that they lacked sufficient knowledge about the behavioral risks that led to their seroconversion; that their decision to engage in unsafe sex was because of negative personal affect; and that they "trusted the wrong person." We link these findings to prevention and suggest that gay and bisexual men who use substances for recreational purposes will benefit from prevention efforts designed to address issues of gay and bisexual men rather than substance-using men. PMID- 19805803 TI - Experiencing out-of-hospital cardiac arrest: significant others' lifeworld perspective. AB - When patients suffer out-of-hospital cardiac arrests (OHCA), significant others find themselves with no choice about being there. After the event they are often left with unanswered questions about the life-threatening circumstances, or the patient's death, or emergency treatment and future needs. When it is unclear how the care and the event itself will affect significant others' well-being, prehospital emergency personnel face ethical decisions. In this article we describe the experiences of significant others present at OHCA, focusing on ethical aspects and values. Using a lifeworld phenomenological approach, 7 significant others were interviewed. The essence of the phenomenon of OHCA can be stated as unreality in the reality, which is characterized by overwhelming responsibility. The significant others experience inadequacy and limitation, they move between hope and hopelessness, and they struggle with ethical considerations and an insecurity about the future.The study findings show how significant others' sense of an OHCA situation, when life is trembling, can threaten values deemed important for a good life. PMID- 19805804 TI - Relationships between humor, subversion, and genuine connection among persons with severe mental illness. AB - Although humor has been linked to resilience among many populations, little is known regarding the role of humor in the coping of individuals with severe mental illness (SMI). In this study, a series of interviews focused on humor was completed by 15 individuals with SMI, with narratives analyzed using grounded theory methods.The marginalized and stigmatized social position occupied by persons with SMI was found to affect both the use and meanings of humor. Humor was described as being the subject of clinical scrutiny. Humor was also emphasized as a means of subverting power differentials revolving around the identity of SMI with, for many, the primary goal being the development of "real" and genuine connections with service providers. PMID- 19805805 TI - Family presence during resuscitation and invasive procedures: the nurse experience. AB - The purpose of this study was to understand the experiences of nurses who participate with families during resuscitation and invasive procedures in the hospital. Seventeen nurses in a large metropolitan area in the northeastern United States were interviewed about their thoughts and feelings about participating with families during these procedures. Conversations were audiotaped and transcribed verbatim for analysis. Transcripts were analyzed using van Manen's technique of isolating thematic statements. Four main themes were identified from the data: forging a connection, engaging the family, transition to acceptance, and a cautious approach.The experience of participating in family presence during resuscitation or invasive procedures can be a positive experience for the nurse by forging a connection with family.The experience for the nurse might include a transition to acceptance and a committed change in practice. PMID- 19805806 TI - I've never not had it so I don't really know what it's like not to: nondifference and biographical disruption among children and young people with cystic fibrosis. AB - The relevance of biographical disruption and loss of self for children and young people is unclear, particularly in cases of congenital illness such as cystic fibrosis, where no prior period of wellness, stability, or perceived normality might exist. We explored the meaning, importance, and forms of maintenance of ideas of normality among 32 children and young people with cystic fibrosis. We examine the ways in which normalcy is produced, maintained, and threatened, and discuss the implications for the applicability and relevance of these traditional sociological concepts. Analysis of children's and young people's accounts resulted in a conceptualization of four forms of normalcy based on personal and social definitions and audiences. Biographical disruption appeared relevant but in a more nuanced form than its usual conceptualization when applied to adult populations. Maintaining normality within the family resulted in continual biographical revision in anticipation of future illness trajectory and life course. PMID- 19805807 TI - From trauma to PTSD: beliefs about sensations, symptoms, and mental illness. AB - Despite the ongoing debate about its legitimacy as a diagnostic entity, posttraumatic stress disorder (PTSD) continues to be the prevailing model for posttrauma suffering in Western societies. In this article, we examine how U.S. veterans came to conceptualize their posttrauma suffering as reflecting the mental disorder of PTSD. We describe the criteria veterans used to evaluate the potential clinical importance of their posttrauma reactions, and the process by which they came to label these reactions as PTSD. Difficulties that veterans experienced in evaluating their posttrauma reactions and in labeling them as PTSD mirror issues that have been raised about the diagnosis of PTSD in the professional literature. PMID- 19805808 TI - Navigating between illness paradigms: treatment seeking by Samoan people in Samoa and New Zealand. AB - There are substantial Samoan communities in New Zealand. Most Samoan people use both palagi (Western) and traditional Samoan health care. Western practitioners sometimes comment on Samoan patients' seemingly erratic patterns of seeking and using health care. Within a larger study on knowledge and the use of antibiotics, we carried out semistructured interviews with 31 Samoan people in Samoa and New Zealand.Accounts of participants' responses to illness and patterns of treatment seeking reveal how Samoan people draw on traditional and Western models of treatment and healing resources. Samoan people are very active interpreters of illness symptoms, using (at least) the two illness paradigms they know of to make sense of symptoms.These paradigms and systems of treatment are sometimes used experimentally, to determine whether illnesses are Samoan or Western. Symptom interpretation and decision making about treatment are done at the family level as well as the individual level. The individual might hold a particular view of what symptoms mean and what to do about them, but might not act on this if overruled or persuaded by other family members. PMID- 19805809 TI - Making it all normal: the role of the internet in problematic pregnancy. AB - Women are actively encouraged to educate themselves about pregnancy from formal sources (e.g., information leaflets, antenatal classes, books). In addition, informal stories of pregnancy and birth are routinely told between women. However, increased prenatal testing means that more fetuses are diagnosed with abnormalities, shifting the information requirements during pregnancy. Traditional sources of information cannot cover all possible outcomes, and the Internet is beginning to fill this gap. In this article, we draw from interviews about experiences of antenatal screening and pregnancy to explore how the Internet provides a unique resource for problematic pregnancies. It allows access to information about rarer conditions beyond standard pregnancy texts, as well as personal narratives about conditions. Learning how others have coped or are coping in similar situations can help alleviate feelings of isolation, and also places women back in a familiar territory of shared pregnancy narratives. PMID- 19805810 TI - Adjusting to life after esophagectomy: the experience of survivors and carers. AB - Following surgery for esophageal cancer, patients can experience complex physical, social, and emotional changes. Investigation of these challenges, particularly from the perspective of the patient and his or her carer, has been limited. The current study explored the emotional and cognitive experiences of esophageal cancer survivors and those of their carers, using focus groups conducted with members of a patient support group. Analysis of the patients' data yielded three themes: coping with a death sentence, adjusting to and accepting an altered self, and the unique benefits of peer support. Analysis of the carers' data also yielded three themes: the carer as buffer, representations of recovery and recurrence, and normalizing experiences through peer support. Esophageal cancer patients and their carers require holistic support in their efforts to adjust to the social, emotional, and physical consequences of esophagectomy. Peers could be an effective channel for the support of patients and carers. PMID- 19805811 TI - Understanding the ecological validity of neuropsychological testing using an ethnographic approach. AB - Neurocognitive impairment is a defining and disabling feature of schizophrenia and other physical disorders. Most of our understanding about neurocognitive deficits comes from laboratory-based testing in research protocols. There has been little research using direct behavioral community observation over a prolonged period to understand the association of daily functioning with cognitive performance.The purpose of this study was to develop an observational method that could be replicated by researchers interested in viewing cognitive deficits in vivo, and then comparing this data to laboratory measures to affirm the ecological validity of those measures.The eight-step method explained here was developed from the targeted ethnographic study of 10 persons with schizophrenia. Obtaining real world context with this method will help to increase the generalizability of effective cognitive treatments, create improved interventions for this population, and bring into greater relief the coping and compensatory strategies already used by individuals to complete daily tasks. PMID- 19805812 TI - Toward an agenda for evaluation of qualitative research. AB - Evaluation is essential for research quality and development, but the diversity of traditions that characterize qualitative research suggests that general checklists or shared criteria for evaluation are problematic. We propose an approach to research evaluation that encourages reflexive dialogue through use of an evaluation agenda. In proposing an evaluation agenda we shift attention from rule-based judgment to reflexive dialogue. Unlike criteria, an agenda may embrace pluralism, and does not request consensus on ontological, epistemological, and methodological issues, only consensus on what themes warrant discussion. We suggest an evaluation agenda-EPICURE-with two dimensions communicated through use of two acronyms.The first, EPIC, refers to the challenge of producing rich and substantive accounts based on engagement, processing, interpretation, and (self )critique. The second-CURE-refers to the challenge of dealing with preconditions and consequences of research, with a focus on (social) critique, usefulness, relevance, and ethics. The seven items of the composite agenda EPICURE are presented and exemplified. Features and implications of the agenda approach to research evaluation are then discussed. PMID- 19805813 TI - A genomewide RNAi screen for genes that affect the stability, distribution and function of P granules in Caenorhabditis elegans. AB - P granules are non-membrane-bound organelles found in the germ-line cytoplasm throughout Caenorhabditis elegans development. Like their "germ granule" counterparts in other animals, P granules are thought to act as determinants of the identity and special properties of germ cells, properties that include the unique ability to give rise to all tissues of future generations of an organism. Therefore, understanding how P granules work is critical to understanding how cellular immortality and totipotency are retained, gained, and lost. Here we report on a genomewide RNAi screen in C. elegans, which identified 173 genes that affect the stability, localization, and function of P granules. Many of these genes fall into specific classes with shared P-granule phenotypes, allowing us to better understand how cellular processes such as protein degradation, translation, splicing, nuclear transport, and mRNA homeostasis converge on P granule assembly and function. One of the more striking phenotypes is caused by the depletion of CSR-1, an Argonaute associated with an endogenous siRNA pathway that functions in the germ line. We show that CSR-1 and two other endo-siRNA pathway members, the RNA-dependent RNA polymerase EGO-1 and the helicase DRH-3, act to antagonize RNA and P-granule accumulation in the germ line. Our findings strengthen the emerging view that germ granules are involved in numerous aspects of RNA metabolism, including an endo-siRNA pathway in germ cells. PMID- 19805814 TI - A Caenorhabditis elegans RNA-directed RNA polymerase in sperm development and endogenous RNA interference. AB - Short interfering RNAs (siRNAs) are a class of regulatory effectors that enforce gene silencing through formation of RNA duplexes. Although progress has been made in identifying the capabilities of siRNAs in silencing foreign RNA and transposable elements, siRNA functions in endogenous gene regulation have remained mysterious. In certain organisms, siRNA biosynthesis involves novel enzymes that act as RNA-directed RNA polymerases (RdRPs). Here we analyze the function of a Caenorhabditis elegans RdRP, RRF-3, during spermatogenesis. We found that loss of RRF-3 function resulted in pleiotropic defects in sperm development and that sperm defects led to embryonic lethality. Notably, sperm nuclei in mutants of either rrf-3 or another component of the siRNA pathway, eri 1, were frequently surrounded by ectopic microtubule structures, with spindle abnormalities in a subset of the resulting embryos. Through high-throughput small RNA sequencing, we identified a population of cellular mRNAs from spermatogenic cells that appear to serve as templates for antisense siRNA synthesis. This set of genes includes the majority of genes known to have enriched expression during spermatogenesis, as well as many genes not previously known to be expressed during spermatogenesis. In a subset of these genes, we found that RRF-3 was required for effective siRNA accumulation. These and other data suggest a working model in which a major role of the RRF-3/ERI pathway is to generate siRNAs that set patterns of gene expression through feedback repression of a set of critical targets during spermatogenesis. PMID- 19805815 TI - DLA-based strategies for cloning insertion mutants: cloning the gl4 locus of maize using Mu transposon tagged alleles. AB - Digestion-ligation-amplification (DLA), a novel adaptor-mediated PCR-based method that uses a single-stranded oligo as the adaptor, was developed to overcome difficulties of amplifying unknown sequences flanking known DNA sequences in large genomes. DLA specifically overcomes the problems associated with existing methods for amplifying genomic sequences flanking Mu transposons, including high levels of nonspecific amplification. Two DLA-based strategies, MuClone and DLA 454, were developed to isolate Mu-tagged alleles. MuClone allows for the amplification of subsets of the numerous Mu transposons in the genome, using unique three-nucleotide tags at the 3' ends of primers, simplifying the identification of flanking sequences that cosegregate with mutant phenotypes caused by Mu insertions. DLA-454, which combines DLA with 454 pyrosequencing, permits the efficient cloning of genes for which multiple independent insertion alleles are available without the need to develop segregating populations. The utility of each approach was validated by independently cloning the gl4 (glossy4) gene. Mutants of gl4 lack the normal accumulation of epicuticular waxes. The gl4 gene is a homolog of the Arabidopsis CUT1 gene, which encodes a condensing enzyme involved in the synthesis of very-long-chain fatty acids, which are precursors of epicuticular waxes. PMID- 19805816 TI - Comparison of mating designs for establishing nested association mapping populations in maize and Arabidopsis thaliana. AB - The nested association mapping (NAM) strategy promises to combine the advantages of linkage mapping and association mapping. The objectives of my research were to (i) investigate by computer simulations the power and type I error rate for detecting quantitative trait loci (QTL) with additive effects using recombinant inbred line (RIL) populations of maize derived from various mating designs, (ii) compare these estimates to those obtained for RIL populations of Arabidopsis thaliana, (iii) examine for both species the optimum number of inbreds used as parents of the NAM populations, and (iv) provide on the basis of the results of these two model species a general guideline for the design of NAM populations in other plant species. The computer simulations were based on empirical data of a set of 26 diverse maize inbred lines and a set of 20 A. thaliana inbreds both representing a large part of the genetic diversity of the corresponding species. I observed considerable differences in the power for QTL detection between NAM populations of the same size but created on the basis of different crossing schemes. This finding illustrated the potential to improve the power for QTL detection without increasing the total resources necessary for a QTL mapping experiment. Furthermore, my results clearly indicated that it is advantageous to create NAM populations from a large number of parental inbreds. PMID- 19805817 TI - Deleting the 14-3-3 protein Bmh1 extends life span in Saccharomyces cerevisiae by increasing stress response. AB - Enhanced stress response has been suggested to promote longevity in many species. Calorie restriction (CR) and conserved nutrient-sensing target of rapamycin (TOR) and protein kinase A (PKA) pathways have also been suggested to extend life span by increasing stress response, which protects cells from age-dependent accumulation of oxidative damages. Here we show that deleting the yeast 14-3-3 protein, Bmh1, extends chronological life span (CLS) by activating the stress response. 14-3-3 proteins are highly conserved chaperone-like proteins that play important roles in many cellular processes. bmh1Delta-induced heat resistance and CLS extension require the general stress-response transcription factors Msn2, Msn4, and Rim15. The bmh1Delta mutant also displays a decreased reactive oxygen species level and increased heat-shock-element-driven transcription activity. We also show that BMH1 genetically interacts with CR and conserved nutrient-sensing TOR- and PKA-signaling pathways to regulate life span. Interestingly, the level of phosphorylated Ser238 on Bmh1 increases during chronological aging, which is delayed by CR or by reduced TOR activities. In addition, we demonstrate that PKA can directly phosphorylate Ser238 on Bmh1. The status of Bmh1 phosphorylation is therefore likely to play important roles in life-span regulation. Together, our studies suggest that phosphorylated Bmh1 may cause inhibitory effects on downstream longevity factors, including stress-response proteins. Deleting Bmh1 may eliminate the inhibitory effects of Bmh1 on these longevity factors and therefore extends life span. PMID- 19805818 TI - Identification of novel genes that mediate innate immunity using inbred mice. AB - Innate immunity is the first line of defense against microbial infections. Although polymorphisms in toll-like receptors (TLRs) and downstream signaling molecules (CD14, TLR2, TLR4, TLR5, and IRAK4) affect the innate immune response, these variants account for only a portion of the ability of the host to respond to bacteria, fungi, and viruses. To identify other genes involved in the innate immune response, we challenged 16 inbred murine strains with lipopolysaccharide (LPS) systemically and measured serum concentrations of pro-inflammatory cytokines IL-1beta, IL-6, and TNFalpha, and the chemokine KC 6 hr post-treatment. Loci that segregate with strain phenotypes were identified by whole genome association (WGA) mapping of cytokine concentrations. Published gene expression profiles and quantitative trait loci (QTL) were then utilized to prioritize loci and genes that potentially regulate the host response to LPS. Sixteen loci were selected for further investigation by combining WGA analysis with previously published QTL for murine response to LPS or gram negative bacteria. Thirty-eight genes within these loci were then selected for further investigation on the basis of the significance of the identified locus, transcriptional response to LPS, and biological plausibility. RNA interference-mediated inhibition of 4 of 38 candidate genes was shown to block the production of IL-6 in J774A.1 macrophages. In summary, our analysis identified 4 genes that have not previously been implicated in innate immunity, namely, 1110058L19Rik, 4933415F23Rik, Fbxo9, and Ipo7. These genes could represent potential sepsis biomarkers or therapeutic targets that should be further investigated in human populations. PMID- 19805819 TI - Centromere replication timing determines different forms of genomic instability in Saccharomyces cerevisiae checkpoint mutants during replication stress. AB - Yeast replication checkpoint mutants lose viability following transient exposure to hydroxyurea, a replication-impeding drug. In an effort to understand the basis for this lethality, we discovered that different events are responsible for inviability in checkpoint-deficient cells harboring mutations in the mec1 and rad53 genes. By monitoring genomewide replication dynamics of cells exposed to hydroxyurea, we show that cells with a checkpoint deficient allele of RAD53, rad53K227A, fail to duplicate centromeres. Following removal of the drug, however, rad53K227A cells recover substantial DNA replication, including replication through centromeres. Despite this recovery, the rad53K227A mutant fails to achieve biorientation of sister centromeres during recovery from hydroxyurea, leading to secondary activation of the spindle assembly checkpoint (SAC), aneuploidy, and lethal chromosome segregation errors. We demonstrate that cell lethality from this segregation defect could be partially remedied by reinforcing bipolar attachment. In contrast, cells with the mec1-1 sml1-1 mutations suffer from severely impaired replication resumption upon removal of hydroxyurea. mec1-1 sml1-1 cells can, however, duplicate at least some of their centromeres and achieve bipolar attachment, leading to abortive segregation and fragmentation of incompletely replicated chromosomes. Our results highlight the importance of replicating yeast centromeres early and reveal different mechanisms of cell death due to differences in replication fork progression. PMID- 19805820 TI - The genetic basis of phenotypic adaptation II: the distribution of adaptive substitutions in the moving optimum model. AB - We consider a population that adapts to a gradually changing environment. Our aim is to describe how ecological and genetic factors combine to determine the genetic basis of adaptation. Specifically, we consider the evolution of a polygenic trait that is under stabilizing selection with a moving optimum. The ecological dynamics are defined by the strength of selection, sigma, and the speed of the optimum, v; the key genetic parameters are the mutation rate Theta and the variance of the effects of new mutations, omega. We develop analytical approximations within an "adaptive-walk" framework and describe how selection acts as a sieve that transforms a given distribution of new mutations into the distribution of adaptive substitutions. Our analytical results are complemented by individual-based simulations. We find that (i) the ecological dynamics have a strong effect on the distribution of adaptive substitutions and their impact depends largely on a single composite measure gamma=v/(sigmaThetaomega(3)), which combines the ecological and genetic parameters; (ii) depending on gamma, we can distinguish two distinct adaptive regimes: for large gamma the adaptive process is mutation limited and dominated by genetic constraints, whereas for small gamma it is environmentally limited and dominated by the external ecological dynamics; (iii) deviations from the adaptive-walk approximation occur for large mutation rates, when different mutant alleles interact via linkage or epistasis; and (iv) in contrast to predictions from previous models assuming constant selection, the distribution of adaptive substitutions is generally not exponential. PMID- 19805822 TI - Pediatric constraint-induced movement therapy is associated with increased contralateral cortical activity on functional magnetic resonance imaging. AB - The mechanism behind constraint-induced movement therapy (constraint therapy) success is unknown. Study objectives were to evaluate cortical change after modified constraint therapy and explore a novel approach to quantify developmental disregard. Five participants underwent modified constraint therapy. Functional magnetic resonance imaging (MRI) and clinical measures were done pretreatment and posttreatment. Developmental disregard indices were calculated. Four participants showed clinical improvement posttreatment. Functional MRI laterality indices were variable pretreatment and exclusively contralateral among participants posttreatment. The disregard index range was -12.9 to 62.6 among participants. Disregard indices were correlated with change scores after treatment on the Pediatric Motor Activity Log amount of use domain (r = .93, P = .02), Assisting Hand Assessment (r = .93, P = .02), and grip strength (r = .92, P = .03). Study results suggest that a shift to or persistence of contralateral cortical activity for affected hand movement is important for constraint therapy mechanism of action; and developmental disregard may be a predictor of positive response to treatment. PMID- 19805821 TI - Efficient transposition of Tol2 in the mouse germline. AB - Insertional mutagenesis screens play an integral part in the annotating of functional data for all sequenced genes in the postgenomic era. Chemical mutagenesis screens are highly efficient but identifying the causative gene can be a laborious task. Other mutagenesis platforms, such as transposable elements, have been successfully applied for insertional mutagenesis screens in both the mouse and rat. However, relatively low transposition efficiency has hampered their use as a high-throughput forward genetic mutagenesis screen. Here we report the first evidence of germline activity in the mouse using a naturally active DNA transposon derived from the medaka fish called Tol2, as an alternative system for high-throughput forward genetic mutagenesis screening tool. PMID- 19805823 TI - The T allele of the 677C>T polymorphism of methylenetetrahydrofolate reductase gene is associated with an increased risk of ischemic stroke in Polish children. AB - Ischemic stroke is a very rare and multifactorial disease in children. The aim of the study was to analyze the relationship between the methylenetetrahydrofolate reductase (MTHFR) 677C>T polymorphism and stroke in Polish children and to observe whether there is any significant transmission of MTHFR alleles from heterozygous parents to their affected offspring. We analyzed 64 patients with stroke, 122 parents, and 59 healthy children. The MTHFR polymorphism was genotyped using polymerase chain reaction (PCR)-restriction fragment length polymorphism. The T allele was more frequent in the stroke group (38%) than in controls (25%, P = .029, odds ratio = 1.84). We also found higher frequency of T allele in male patients compared to male controls (46% vs. 25%, P = .009, odds ratio = 2.53). The number of T allele carriers was again more prevalent in boys with stroke (71%) than in healthy boys (45%, P = .023, odds ratio = 3.09). The T allele was significantly transmitted in male patients (P < .019). We conclude that the MTHFR 677C>T polymorphism may be considered as a genetic risk factor of childhood stroke, especially in boys. PMID- 19805824 TI - Attention-deficit hyperactivity disorder (ADHD) and tuberous sclerosis complex. AB - The neurobiological basis of attention-deficit hyperactivity disorder (ADHD) in tuberous sclerosis complex is still largely unknown. Cortical tubers may disrupt several brain networks that control different types of attention. Frontal lobe dysfunction due to seizures or epileptiform electroencephalographic discharges may perturb the development of brain systems that underpin attentional and hyperactive functions during a critical early stage of brain maturation. Comorbidity of attention-deficit hyperactivity disorder (ADHD) with mental retardation and autism spectrum disorders is frequent in children with tuberous sclerosis. Attention-deficit hyperactivity disorder (ADHD) may also reflect a direct effect of the abnormal genetic program. Treatment of children with tuberous sclerosis complex with combined symptoms of attention-deficit hyperactivity disorder (ADHD) and epilepsy may represent a challenge for clinicians, because antiepileptic therapy and drugs used to treat attention deficit hyperactivity disorder (ADHD) may aggravate the clinical picture of each other. PMID- 19805825 TI - SURF-1 gene mutation associated with leukoencephalopathy in a 2-year-old. AB - Mutations in the nuclear SURF-1 gene lead directly to cytochrome-c oxidase deficiency, the most common respiratory chain defect in Leigh syndrome, a neurodegenerative mitochondrial disease involving the deep gray matter and brain stem. We describe the second documented case in the literature to have a SURF-1 mutation presenting with diffuse leukodystrophy, adding to the growing number of cases of mitochondrial syndromes presenting with white matter disease. We examine magnetic resonance imaging (MRI) findings, which suggest that high-grade cytotoxic edema on diffusion-weighted imaging may be a helpful diagnostic feature in differentiating mitochondrial leukodystrophy from other, more common leukodystrophies. We show how MRI white matter findings may progress to include the brain stem, suggesting that a leukodystrophy due to respiratory chain defects can precede more classic Leigh syndrome deep gray matter radiographic findings. PMID- 19805826 TI - Inflammatory myofibroblastic tumor involving lung and brain in a 10-year-old boy: a case report. AB - Inflammatory myofibroblastic tumors are rare tumors of unknown etiology, composed of proliferating myofibroblasts and accompanying lymphoplasmacytic infiltration. A 10-year-old boy who developed inflammatory myofibroblastic tumors in the lung as well as the brain is described. Surgery and radiation therapy were not feasible. The authors review the current literature and treatment options. PMID- 19805827 TI - Conflict of interest in peer-reviewed medical journals: a policy statement of the World Association of Medical Editors (WAME). PMID- 19805828 TI - Correspondence on ''effect of acupuncture on the brain in children with spastic cerebral palsy using functional neuroimaging (fMRI)''. PMID- 19805833 TI - Elevated admission serum creatinine predicts poor myocardial blood flow and one year mortality in ST-segment elevation myocardial infarction patients undergoing primary percutaneous coronary intervention. AB - BACKGROUND: Outcomes after percutaneous coronary intervention (PCI) for patients with acute myocardial infarction (AMI) complicated by renal insufficiency have been well described. However, data regarding admission serum creatinine and coronary and myocardial flow are scant. The aims of this study are to evaluate the effects of admission serum creatinine on coronary blood flow and prognosis in ST-segment elevation myocardial infarction (STEMI) patients undergoing primary PCI. METHODS: A total of 495 patients undergoing primary PCI for STEMI within 12 hours after symptom onset were studied. Patients were divided into two groups according to admission serum creatinine level: 1) elevated serum creatinine group (elevated group, serum creatinine > or = 1.3 mg/dl), and 2) normal serum creatinine group (normal group, serum creatinine < 1.3 mg/dl). RESULTS: Elevated serum creatinine was observed in 86 patients. Univariate analyses showed statistical differences between normal and elevated serum creatinine groups in age, gender, number of diseased vessels, hypertension, previous MI, serum creatinine level and Killip's grades on presentation. Corrected TIMI frame count (CTFC) in the elevated group was more than in the normal group, and TIMI myocardial perfusion grades (TMPG) 0-1 were more frequent (5.6% and 12.8%, p < 0.05). Elevated admission serum creatinine was an independent predictor of poor myocardial perfusion (adjusted relative risk [RR] 3.93; 95% confidence interval [CI] 1.13-6.84) and a higher rate of 1-year mortality in STEMI patients undergoing primary PCI (adjusted RR 1.41; 95% CI 1.24-2.69]). CONCLUSIONS: The elevated admission serum creatinine levels are associated with impaired myocardial flow and poor prognosis in STEMI patients undergoing primary PCI. PMID- 19805834 TI - Renal insufficiency and prognosis after primary PCI: still bad after all these years. PMID- 19805835 TI - Peripheral plaque volume changes pre- and post-rotational atherectomy followed by directional plaque excision: assessment by intravascular ultrasound and virtual histology. AB - Atherectomy in the infra-inguinal peripheral vascular bed may be an effective alternative to the balloon and stent-based approach. The change in plaque volume and composition with rotational atherectomy and directional plaque excision has not been studied. We performed rotational atherectomy (RA) followed by adjunctive plaque excision (PE) in 8 patients with infra-inguinal lesions. Lesions were assessed by quantitative angiography and intravascular ultrasound (IVUS) with virtual histology (VH) pretreatment, post-RA, and post-PE. Paired t-tests were used to assess the change in plaque volume, luminal area and dimensions and overall vessel size. Total plaque volume decreased by 24% (347 +/- 118 to 264 +/- 85 mm(3); p = 0.03), which resulted in a lumen increase of 66% (7.08 +/- 3 to 11.7 +/- 2.8 mm(2); p = 0.006). This was mainly due to PE compared to RA. This was achieved without any significant increase in the overall vessel area (27.4 +/ 6.3 to 28 +/- 6.7 mm(2); p = 0.42). The effect on minimal and maximal luminal diameters was synergistic between RA and PE. Fibrotic and fibrofatty plaque were amenable to PE, whereas necrotic core and dense calcium were less responsive to either atherectomy technique. In conclusion, RA followed by PE result in significant improvements in plaque volume and luminal area and diameter primarily by removing plaque rather than vessel expansion. This is mainly attributable to the removal of fibrotic and fibrofatty plaque. PMID- 19805836 TI - Ethnic minorities exhibit reduction in door-to-balloon time comparable to whites with a newly-implemented emergency physician activation protocol (Code-STEMI). AB - BACKGROUND: Studies have shown significant differences in door-to-balloon times (D2B) for ethnic minority patients (minorities) undergoing angioplasty for ST elevation myocardial infarction (STEMI) compared to white patients (white). In this study, we evaluated the D2B for these groups before and after modification of the emergency protocol for STEMI. METHODS: We compared D2B for 51 consecutive STEMIs during 2006, (serial activation protocol, SAP) with D2B times for 72 consecutive STEMI patients during 2007 when a "Code STEMI" (concurrent activation) protocol was instituted. Outcomes were D2B times in whites versus minorities, pre- and post-Code-STEMI, length of stay (LOS) and peak troponin I levels. RESULTS: The median D2B time in the SAP group was 113 (whites) vs. 122 (minorities) minutes (p = 0.324), as compared to 74 (whites) vs. 78 (minorities) minutes (p = 0.324) in the Code STEMI group. The D2B for both groups was significantly reduced (p < 0.0001) with the use of Code STEMI. The median peak troponin I in the SAP group was 97 ng/mL (whites) vs. 78 ng/mL (minorities) (p = 0.084), as compared to 54 ng/mL (whites) vs. 29 ng/mL (minorities) (p = 0.084) for the Code STEMI group. LOS was 4.88 days (whites) vs. 4.39 days (minorities) (p = 0.84) in the SAP group, as compared to 3.7 days (whites) vs. 3.4 days (minorities) (p = 0.84) for the Code STEMI group, a significant change (p = 0.012) for both groups. CONCLUSION: No ethnic disparity was observed in the mean D2B time, LOS and peak troponin I levels between whites and minorities; both groups demonstrated comparable improvement in all outcomes evaluated. PMID- 19805837 TI - Concurrent assessment of epicardial coronary artery stenosis and microvascular dysfunction using diagnostic endpoints derived from fundamental fluid dynamics principles. AB - BACKGROUND: Simultaneously measured pressure and flow distal to coronary stenoses can be combined, in conjunction with anatomical measurements, to assess the status of both the epicardial and microvascular circulations. METHODS AND RESULTS: Assessments of coronary hemodynamics were performed using fundamental fluid dynamics principles. We hypothesized that the pressure-drop coefficient (CDPe; trans-stenotic pressure drop divided by the dynamic pressure in the distal vessel) correlates linearly with epicardial and microcirculatory resistances concurrently. In 14 pigs, simultaneous measurements of distal coronary arterial pressure and flow were performed using a dual sensor-tipped guidewire in the setting of both normal and disrupted microcirculation, with the presence of epicardial coronary lesions of lt; 50% area stenosis (AS) and > 50% AS. The CDPe progressively increased from lesions of < 50% AS to > 50% AS and had a higher resolving power (45 +/- 22 to 193 +/- 140 in normal microcirculation; 248 +/- 137 to 351 +/- 140 in disrupted microcirculation) as compared to fractional flow reserve (FFR) and coronary flow reserve (CFR). Strong multiple linear correlation was observed for CDPe with combined FFR and CFR (r = 0.72; p < 0.0001). Further, the ratio of maximum pressure drop coefficient evaluated at the site of stenosis and its theoretical limiting value of minimum cross-sectional area was also able to distinguish different combinations of coronary artery diseases. CONCLUSIONS: The CDPe can be readily obtained during routine pressure and flow measurements during cardiac catheterization. It is a promising clinical diagnostic parameter that can independently assess the severity of epicardial stenosis and microvascular impairment. PMID- 19805838 TI - ST-elevation myocardial infarction mortality in a major academic center "on-" versus "off-" hours. AB - BACKGROUND: A higher mortality rate for weekend myocardial infarction (MI) admissions has been reported and attributed to the lower availability of primary percutaneous coronary intervention (PCI) during off-hours. However, the data are conflicting and, furthermore, inapplicable to hospitals where primary PCI is invariably performed. METHODS: This study was conducted in a tertiary hospital where primary PCI is routinely performed in all patients with ST-elevation myocardial infarction (STEMI). Patients admitted during on-hours (Monday through Friday 7 am-7 pm) where compared to off-hours patients (including weekends). The primary endpoint of in-hospital mortality, cardiogenic shock and recurrent MI was examined. A second analysis that excluded STEMI transfers, in-hospital mortality and reperfusion times was examined. RESULTS: Between 2003 and 2007, 747 STEMI patients (46% on-hours vs. 56% off-hours) underwent primary PCI. Demographic characteristics were similar between on- and off-hours groups. However, off-hours STEMI admissions had significantly greater in-hospital mortality rates (8% vs. 3.7%; p = 0.01) and higher rates of cardiogenic shock (37% vs. 24%; p = 0.0001). Admission arrival time was an independent predictor of in-hospital mortality (hazard ratio [HR] 3.98, 95% confidence interval [CI] 1.10-14.38; p = 0.035). Longer door-to-balloon times (DTB) were observed during off-hours (134 vs. 109 minutes; p < 0.0001), even after excluding the transfer population (63 vs. 89 minutes; p < 0.0001). CONCLUSION: Higher rates of in-hospital mortality and cardiogenic shock may be expected in STEMI patients admitted during off-hours, even when primary PCI is performed. Longer DTB times during off-hours may partially explain our findings. Strategies to optimize reperfusion time during off-hours, including perhaps a 24/7 in-house "STEMI team" may be necessary. PMID- 19805839 TI - On and off: the dilemma of managing STEMI after hours. PMID- 19805840 TI - Long-term safety and efficacy of sirolimus-eluting stents in Japanese patients: a single-center cohort study. AB - BACKGROUND: The long-term safety and efficacy of sirolimus-eluting stents (SES) remain uncertain in real practice in Japan. METHODS: We used a hospital-based cohort (n = 6,562) comprising all the new patients who had visited our hospital between 2004 and 2007 to investigate the mortality and morbidity after SES or bare-metal stent (BMS) implantation. RESULTS: Of the total, coronary artery disease was observed in 822 patients (12.5%), and SES or BMS were implanted in 208 and 167 patients, respectively. Patients receiving SES more often had diabetes but less often presented with acute myocardial infarction (MI). Median follow-up periods were 815 and 894 days for SES and BMS, respectively (p = 0.305). Intravascular ultrasound (IVUS) was used at a high rate (> 90%) in both groups, and maximum pressure inflation for SES was high at approximately 18 atm. The unadjusted cumulative incidence of all-cause death and major adverse cardiac events (MACE) (cardiac death, MI or target vessel revascularization) at 2 years was 4.3% versus 7.2% (p = 0.233) and 16.3% versus 32.9% (p < 0.001), respectively. In multivariate analysis, all-cause mortality was similar between SES (hazard ratio [HR] 0.981, 95% confidence interval [CI] 0.366-2.632) and BMS, but SES significantly reduced MACE (HR 0.468, 95% CI 0.280-0.784). Definite stent thrombosis set by the Academic Research Consortium was not observed in either group, and the incidence of cerebral hemorrhage was 0.5% in SES and 1.2% in BMS, respectively (p = 0.588). CONCLUSIONS: SES used in real-world settings appeared to be safe and significantly associated with a lower risk of adverse events at long-term follow up in Japan, especially utilizing a careful stent deployment technique including high-pressure inflation and IVUS use. PMID- 19805841 TI - Provisional TAP-stenting strategy to treat bifurcated lesions with drug-eluting stents: one-year clinical results of a prospective registry. AB - OBJECTIVE: To assess the clinical outcome of unselected patients undergoing drug eluting stent (DES) implantation on bifurcated lesions using a "provisional T And small Protrusion (TAP)" stenting strategy. METHODS: Consecutive patients undergoing DES implantation on one major bifurcation lesion were treated by main vessel (MV) stenting, followed (if needed) by side-branch (SB) rewiring (with a "pullback" technique) and kissing balloon. SB stenting was performed according to the TAP-technique in selected cases. The endpoint of the study was a 12-month incidence of major adverse cardiac events (MACE) defined as cardiac death, myocardial infarction (MI), stent thrombosis and target vessel revascularization (TVR). RESULTS: The study population included 266 patients (9% unprotected left main). Only 19 patients (7.1%) (with more complex angiographic features) received stents in both the MV and SB using the TAP-technique. Overall, 22 (8.2%) patients had MACE at 1 year. Observed, non-hierarchical MACE were: 1 (0.4%) cardiac death, 11 (4.1%) MI, 2 probable stent thromboses and 12 (4.5%) TVRs. Postprocedural troponin T increase and adverse events up to 12 months were similar between patients treated by MV stenting only or double stenting. CONCLUSIONS: In unselected patients undergoing DES implantation on bifurcated lesions, a provisional TAP-stenting strategy (with a low rate of SB stenting) appears to be safe and effective. PMID- 19805842 TI - "Bailing out" side branches: technique trumps technology, round 1. PMID- 19805843 TI - The confluent balloon technique--two cases illustrating a novel method to achieve rapid wire crossing of chronic total occlusion during retrograde approach percutaneous coronary intervention. AB - We describe two cases of a retrograde approach to treat a chronic total occlusion (CTO) of the right coronary artery in which we were initially unable to achieve wire crossing. A novel technique of pushing both antegrade and retrograde balloons into the CTO lesion so that the balloons overlapped each other during inflation created a new confluent subintimal space allowing easy, reliable and rapid passing of the retrograde wire into the antegrade true lumen. This novel "confluent balloon" method is recommended to other operators who may struggle with wire crossing during a retrograde approach to CTOs. PMID- 19805844 TI - Retrograde PCI: what will they think of next? PMID- 19805845 TI - Management of radial and brachial artery perforations during transradial procedures--a practical approach. AB - We present a practical approach to the management of iatrogenic radial and brachial artery perforations during transradial catheterization and interventions. These perforations were treated successfully without abandoning the procedure in 15 patients. PMID- 19805846 TI - Anaphylactoid reactions to radiocontrast agents: prevention and treatment in the cardiac catheterization laboratory. AB - The use of iodinated contrast agents for angiography dates back to the 1920s. The initial prototype has undergone modifications to reduce the toxicity and discomfort associated with the early contrast molecules. More importantly, these changes have dramatically decreased the rate and risk for severe adverse reactions such as hypersensitivity and anaphylaxis. With over 15 million contrast requiring procedures performed annually in the United States, it is important to understand the risk factors, pathogenesis, diagnosis, prevention and treatment of contrast-induced anaphylactoid reactions. Reviews of adverse reactions are sparse in the cardiology literature, except for a landmark review in 1995 by Goss et al, which has served as the only practice guideline to date for cardiologists. In this report, we review the most recent literature to provide a guide for the general and interventional cardiologist in regards to the pretreatment and management of contrast-related reactions specifically in the cardiac catheterization laboratory. PMID- 19805847 TI - Detection of atherosclerotic progression with rupture of degenerated in-stent intima five years after bare-metal stent implantation using optical coherence tomography. AB - In-stent intimal hyperplasia peaks in the early phase (6-12 months) after bare metal stent (BMS) implantation. However, late luminal re-narrowing due to atherosclerotic progression of in-stent intima is reported beyond 4 years. We report the optical coherence tomographic findings of a case of late restenosis of BMS 5 years after implantation. Remarkable in-stent intimal growth was observed, demonstrating a heterogeneous appearance including low-intensity areas and accompanied by intimal rupture. These findings were similar to the morphology of ruptured fibroatheroma in the native coronary artery, and suggested that atherosclerotic change in the in-stent intima occurred over the 5 years following BMS implantation. PMID- 19805848 TI - Myocardial infarction due to paradoxical embolism in a patient with large atrial septal defect. AB - We present the case of a patient who presented with acute inferior myocardial infarction and embolic occlusion of the distal left anterior descending and proximal right coronary artery. A large atrial septal defect (ASD) was seen on transesophageal echocardiography and the ASD was closed during the same session as coronary angiography and percutaneous coronary intervention. The presence of embolic or thrombotic occlusions of coronary arteries should prompt interventional cardiologists to look for a patent foramen ovale or ASD and perform percutaneous closure right away. PMID- 19805849 TI - Use of a novel pericardial covered stent to seal an iatrogenic coronary perforation. AB - Iatrogenic coronary perforation complicates 0.1-0.8% of percutaneous coronary interventional (PCI) procedures. The incidence is higher if atheroablative therapy is used. When coronary perforation occurs, it may rapidly result in cardiac tamponade, myocardial infarction or death, hence prompt treatment is required. PTFE-covered stents have been used to seal coronary perforations, but these are bulky devices that lack flexibility, and rapid deployment in calcified or tortuous vessels can be difficult, particularly in emergency situations. Furthermore, difficulties in achieving adequate stent expansion and the prospect of delayed re-endothelialization have led to concern about the increased potential for stent thrombosis or restenosis. We present the first report of the successful use of a novel, highly deliverable pericardial covered stent to treat an iatrogenic coronary perforation during PCI, with angiographic follow up. PMID- 19805850 TI - Rotational atherectomy is useful to treat restenosis lesions due to crushing of a sirolimus-eluting stent implanted in severely calcified lesions: experimental study and initial clinical experience. AB - We have occasionally encountered restenosis due to the crushing of drug-eluting stents (DES) implanted in severely calcified lesions. We aimed to establish the role of rotational atherectomy (RA) in its treatment. At first, we conducted an experimental study and found that the size of the metallic particles generated during RA of stent struts was 5.6 +/- 3.6 mum. We performed RA on the restenosis of the sirolimus-eluting stents implanted in the severely calcified lesions of a 66-year-old male who had received hemodialysis for 13 years. He had restenosis in the proximal and mid-segments of the right coronary artery, and intravascular ultrasound images documented that these stents were crushed by calcified plaque behind them. RA ablated both crushed stent struts and the calcified lesions behind them, and there was no hemodynamic derangement during the procedure. Maximum dilatation of the lesions was achieved with balloon angioplasty, followed by stent implantation. RA is an effective strategy to treat restenotic lesions resulting from the crushing of DES in severely calcified lesions. PMID- 19805851 TI - Transcatheter valve-in-valve implantation due to severe aortic regurgitation in a degenerated aortic homograft. AB - Transcatheter aortic valve implantation (TAVI) in severe aortic stenosis has proven to be a feasible and effective treatment modality for inoperable patients. Until now, neither aortic regurgitation nor degenerated bioprostheses has been an indication for TAVI. However, this article reports a successful valve-in-valve implantation of a CoreValve aortic valve prosthesis through the right subclavian artery in a case of severe aortic regurgitation within a degenerated aortic homograft. The case exemplifies the possibilities of expanding the indications for TAVI, as well as other vascular access options than the femoral arteries. PMID- 19805852 TI - Embolized stent in the coronary circulation "without riding the wire" and its novel management strategy. AB - We report a unique technique for retrieval of a coronary stent, embolized during percutaneous coronary intervention (PCI), using conventional angioplasty wires and angioplasty balloon catheters. In this case, the stent embolized into the distal left main and proximal left circumflex arteries. As the guide catheter and percutaneous coronary angioplasty (PTCA) wire were withdrawn from the coronary arteries, the stent was freely floating in the coronary circulation "without riding the wire". It was managed by re-entry of the Intermediate PTCA wire through the stent with the help of an over-the-wire balloon and was successfully deployed across the lesion. Stent embolization, although a very rare event, may lead to devastating consequences. This technique has been demonstrated for the first time and employs readily available tools that are familiar to all operators. PMID- 19805853 TI - Anomalous circumflex coronary artery injury caused by mitral annuloplasty: role of 64-multislice computed tomography. PMID- 19805855 TI - Probing the nanoworld. PMID- 19805854 TI - Mastering matter at the nanoscale. PMID- 19805856 TI - Nanotechnology impact on sensors. PMID- 19805857 TI - Nature's building blocks. PMID- 19805858 TI - Quantum phenomena in Nanotechnology. PMID- 19805859 TI - Moore and more progress in electronics and photonics. PMID- 19805861 TI - Big science at the nanoscale. PMID- 19805860 TI - Whither nanomaterials? PMID- 19805862 TI - Bi4Ge3O12 at the onset of pressure-induced amorphization. AB - The crystal structure of tetrabismuth tris(germanate), Bi4Ge3O12 (I43d, Z = 4), is stable to at least 7.30 GPa, as demonstrated by hydrostatic single-crystal X ray diffraction measurements in a diamond anvil cell at room temperature. The highest pressure reached in this study is close to the onset of amorphization at about 8 GPa. The Bi and Ge atoms are located at the 16c (3) and 12a (4) Wyckoff positions, respectively. The compression mainly affects the distorted BiO6 octahedra, while the GeO4 tetrahedra are relatively rigid. When compared with the values obtained under ambient conditions, the long Bi-O distances decrease with increasing pressure, while the short Bi-O distances do not change. PMID- 19805863 TI - EuI2, a low-temperature europium(II) iodide phase. AB - Light-yellow europium(II) diiodide, prepared by the low-temperature reaction of europium and ammonium iodide in liquid ammonia at 200 K and characterized by high resolution X-ray powder diffraction, represents a new phase for EuI2 that adopts an orthorhombic Pnma structure with all three atoms lying on 4c positions (.m.). It is isotypic with SrI2(IV). Temperature-dependent X-ray measurements performed to investigate the thermal stability of the new phase show that it decomposes irreversibly to amorphous material around 673 K. Total-energy density-functional calculations using the generalized gradient approximation suggest this to be the ground-state structure of EuI2. PMID- 19805864 TI - Flux growth and structure of two compounds with the EuIn2P2 structure type, AIn2P2 (A = Ca and Sr), and a new structure type, BaIn2P2. AB - Single crystals of the new Zintl phases AIn2P2 [A = Ca (calcium indium phosphide), Sr (strontium indium phosphide) and Ba (barium indium phosphide)] have been synthesized from a reactive indium flux. CaIn2P2 and SrIn2P2 are isostructural with EuIn2P2 and crystallize in the space group P63/mmc. The alkaline earth cations A are located at a site with 3m symmetry; In and P are located at sites with 3m symmetry. The structure type consists of layers of A2+ cations separated by [In2P2]2- anions that contain [In2P6] eclipsed ethane-like units that are further connected by shared P atoms. This yields a double layer of six-membered rings in which the In-In bonds are parallel to the c axis and to one another. BaIn2P2 crystallizes in a new structure type in the space group P2(1)/m with Z = 4, with all atoms residing on sites of mirror symmetry. The structure contains layers of Ba2+ cations separated by [In2P2]2- layers of staggered [In2P6] units that form a mixture of four-, five- and six-membered rings. As a consequence of this more complicated layered structure, both the steric and electronic requirements of the large Ba2+ cation are met. PMID- 19805865 TI - Disordered structure of ZrW1.8V0.2O7.9from a combined X-ray and neutron powder diffraction study at 530 K. AB - A novel compound, vanadium aliovalent substituted zirconium tungstate, ZrW1.8V0.2O7.9, was prepared with vanadium substituting tungsten rather than the common zirconium substitution. The structure of the high-temperature phase was refined from combined neutron and X-ray powder diffraction data gathered at 530 K. This phase is the disordered centric modification (space group Pa3) and the average crystal structure is similar to that of beta-ZrW2O8. The V atom occupies only a W2 site and charge compensation is achieved through oxygen vacancy, i.e. the oxygen vacancy occurs at only the O4 site. [Atom names follow the established scheme; Evans et al. (1996). Chem. Mater. 8, 2809-2823.]. PMID- 19805866 TI - Lagoden dimethylformamide hemisolvate dihydrate: absolute configuration, dipolar interactions and hydrogen-bonding interactions. AB - Lagoden (L.3H2O, where L is Na+.C20H33O6-; sodium 3beta,16,18-trihydroxy-8,13-epi 9,13-epoxylabdan-15-oate trihydrate) is widely used as an effective haemostatic agent. It has been crystallized from dimethylformamide (DMF) as sodium 3beta,16,18-trihydroxy-8,13-epi-9,13-epoxylabdan-15-oate dimethylformamide hemisolvate dihydrate, Na+.C20H33O6-.0.5C3H7NO.2H2O or L2.DMF.4H2O, and the asymmetric unit contains two of the latter formulation. The four symmetry independent Na+ cations and lagoden anions, one DMF molecule and six of the eight symmetry-independent water molecules assemble into a one-dimensional polymeric structure via dipolar and hydrogen-bonding interactions. The lagoden anions coordinate to the Na+ cations via the carboxylate groups and the two primary hydroxy groups, whereas the secondary OH groups are solely involved in hydrogen bonding. Two of the four symmetry-independent lagoden anions act in a chelating mode, forming seven-membered chelate rings. The absolute structure, based on anomalous dispersion data collected at 130 K with Cu Kalpha radiation, confirms an inverted configuration at chiral centres C8 and C13 (labdane numbering) relative to the labdane skeleton. PMID- 19805867 TI - Hydrogen-bonded three-dimensional network of a lanthanum(III) exocyclic complex with 5,10,15,20-tetra-4-pyridylporphyrin. AB - In the complex diaquatetranitrato[5-(pyridinium-4-yl)-10,15,20-tri-4 pyridylporphyrin]lanthanum(III) 1,2-dichlorobenzene trisolvate, [La(NO3)4(C40H27N8)(H2O)2].3C6H4Cl2, the lanthanum ion is coordinated to one of the peripheral pyridyl substituents of the porphyrin entity. Units of the complex are interlinked to one another in three dimensions by a network of O-H...N, O H...O and N-H...O hydrogen bonds between the water ligands, nitrate ions, and pyridyl and pyridinium groups of adjacent species. This is the first structural report of an exocyclic complex of the tetrapyridylporphyrin ligand with any lanthanide ion and its self-assembly into a three-dimensional architecture sustained by hydrogen bonds. PMID- 19805868 TI - Poly[diaqua(mu4-benzene-1,2-dicarboxylato)(isonicotinato kappa2O,O')gadolinium(III)]. AB - The title neutral polymer, [Gd(C6H4NO2)(C8H4O4)(H2O)2]n, contains an extended two dimensional wave-like lanthanide carboxylate layer decorated by isonicotinate (IN) ligands. The Gd(II) atom is eight-coordinated by four carboxylate O atoms from four benzene-1,2-dicarboxylate (1,2-bdc) ligands, two 1,2-bdc carboxylate O atoms from one chelating IN ligand and two terminal water molecules, forming a bicapped trigonal-prismatic coordination geometry. The wave-like layers are stacked in an ...ABAB... packing mode along the c-axis direction. Strong hydrogen bonding interactions further stabilize the structure of the title compound. PMID- 19805869 TI - Two novel silver(I) coordination polymers: poly[(mu2-2-aminopyrimidine kappa2N1:N3)bis(mu3-thiocyanato-kappa3S:S:S)disilver(I)] and poly[(2-amino-4,6 dimethylpyrimidine-kappaN)(mu3-thiocyanato-kappa3N:S:S)silver(I)]. AB - 2-Aminopyrimidine (L1) and 2-amino-4,6-dimethylpyrimidine (L2) have been used to create the two novel title complexes, [Ag2(NCS)2(C4H5N3)]n, (I), and [Ag(NCS)(C6H9N3)]n, (II). The structures of complexes (I) and (II) are mainly directed by the steric properties of the ligands. In (I), the L1 ligand is bisected by a twofold rotation axis running through the amine N atom and opposite C atoms of the pyrimidine ring. The thiocyanate anion adopts the rare mu3-kappa3S coordination mode to link three tetrahedrally coordinated Ag(I) ions into a two dimensional honeycomb-like 6(3) net. The L1 ligands further extend the two dimensional sheet to form a three-dimensional framework by bridging Ag(I) ions in adjacent layers. In (II), with three formula units in the asymmetric unit, the L2 ligand bonds to a single Ag(I) ion in a monodentate fashion, while the thiocyanate anions adopt a mu3-kappa1N,kappa2S coordination mode to link the AgL2 subunits to form two-dimensional sheets. These layers are linked by N-H...N hydrogen bonds between the noncoordinated amino H atoms and both thiocyanate and pyrimidine N atoms. PMID- 19805870 TI - [Bis(3,5-dimethylpyrazol-1-yl-kappaN2)hydro(pyrazol-1-yl-kappaN2)borato][(3,5 dimethylpyrazol-1-yl-kappaN2)dihydro(pyrazol-1-yl-kappaN2)borato]nickel(II). AB - The title compound, [Ni(C8H12BN4)(C13H18BN6)] or Bp'Tp'Ni(II), where Bp' is (3,5 dimethylpyrazol-1-yl)dihydro(pyrazol-1-yl)borate and Tp' is bis(3,5 dimethylpyrazol-1-yl)hydro(pyrazol-1-yl)borate, contains a divalent Ni(II) centre bound by the chelating N atoms of the polysubstituted pyrazolylborate ligands. It is shown to lack a strong agostic B-H...Ni interaction, implying that the sixth coordination site is unoccupied in the solid state. This square-pyramidal complex is the only known crystal structure where the Ni(II) centre is pentacoordinated while bonded exclusively to pyrazolyl units. This is of interest with respect to electrochemical and catalytic properties. PMID- 19805871 TI - A new organically templated vanadium tellurite: (H2dien)[(VO2)(TeO3)]2.2H2O (dien is diethylenetriamine). AB - A new organically templated vanadium tellurite, poly[2,2'-iminodiethanaminium [hexa-mu2-oxido-tetraoxidoditellurium(IV)divanadium(V)] dihydrate], {(C4H15N3)[Te2V2O10].2H2O}n, features the interconnection of distorted [VO5] trigonal bipyramids by bridging [TeO3] pyramids, leading to a two-dimensional corrugated anionic layer with an interlayer distance of about 13.47 A. The interlayer space is occupied by doubly protonated diethylenetriamine cations (H2dien) and guest water molecules. The two terminal amino groups of H2dien are protonated, while the middle amino group, located on a twofold rotation axis, is not protonated. All the three amino groups and water molecules are involved in hydrogen-bonding interactions. The compound represents a new member in the series (H2am)[(VO2)(TeO3)]2.xH2O, where H2am represents a doubly protonated diamine. Similarities and differences between the structures of members of the series are discussed. PMID- 19805872 TI - Poly[[aqua(4,4'-diazenediyldibenzoato-kappa4O,O':O'',O''')cadmium(II)]: a twofold interpenetrated three-dimensional coordination polymer of PtS topology. AB - In the title coordination compound, [Cd(C14H8N2O4)(H2O)]n, the Cd(II) cation and the coordinated water molecule lie on a twofold axis, whereas the ligand lies on an inversion center. The Cd(II) center is five-coordinated in a distorted square pyramidal geometry by four carboxylate O atoms from four different 4,4' diazenediyldibenzoate (ddb) anions and one water O atom. The three-dimensional frameworks thus formed by the bridging ddb anions interpenetrate to generate a three-dimensional PtS-type network. Additionally, the coordination water molecule and the carboxylate O atom form a hydrogen-bonding interaction, stabilizing the three-dimensional framework structure. PMID- 19805873 TI - (+ac,-ac)-trans-Bis(hinokitiolato)copper(II) and its chloroform disolvate. AB - The complex trans-bis(hinokitiolato)copper(II) [systematic name: trans-bis(3 isopropyl-7-oxocyclohepta-1,3,5-trienolato)copper(II); abbreviated name: trans Cu(hino)2], [Cu(C10H11O2)2], is a biologically active compound. Three polymorphs of this square-planar monomer, all with (+sp,-sp) isopropyl substituents, have been reported previously. A fourth polymorph containing (+ac,-ac) isopropyl groups and its chloroform disolvate, [Cu(C10H11O2)2].2CHCl3, both exhibiting nonmerohedral twinning and with all Cu atoms on centers of crystallographic inversion symmetry, are reported here. One of the differences between all of these polymorphs is the relative conformation of the isopropyl groups with respect to the plane of the molecule. Stacking and Cu...olefin pi distances ranging from 3.214 (4) to 3.311 (2) A are observed, and the chloroform solvent molecules participate in bifurcated C-H...O hydrogen bonds [H...O = 2.26-2.40 A, C...O = 3.123 (5)-3.214 (5) A, C-H...O = 127-151 degrees and O...H...O = 74 degrees]. PMID- 19805874 TI - Poly[[diaqua(mu3-3-nitrophthalato)calcium(II)] monohydrate]. AB - The title 3-nitrophthalate-calcium coordination polymer, {[Ca(C8H3NO6)(H2O)2].H2O}n, crystallizes as a one-dimensional framework. The Ca(II) centre has a distorted pentagonal-bipyramidal geometry, being seven coordinated by five O atoms from three different 3-nitrophthalate groups and by two water molecules, resulting in a one-dimensional zigzag chain along the a-axis direction by the interconnection of the four O atoms from the two carboxylate groups. There is a D3 water cluster composed of the coordinated and the solvent water molecules within such chains. Adjacent chains are aggregated into two dimensional layers via hydrogen bonds in the c-axis direction. The whole three dimensional structure is further stabilized by weak O-H...O hydrogen bonds between the O atoms of the nitro group and the water molecules. PMID- 19805875 TI - Octakis(dimethylammonium) hexa-mu2-chlorido-hexachloridotrinickelate(II) dichloride: a linear trinickel complex with asymmetric bridging. AB - The title compound, (C2H8N)8[Ni3Cl12]Cl2, crystallizes as linear [Ni3Cl12]6- complex anions with inversion symmetry, separated from one another by dimethylammonium cations and noncoordinated chloride ions. The gross structural arrangement of the trinickel complex is as a segment of face-sharing NiCl6 octahedra similar to the (NiCl3)n chains of CsNiCl3-type compounds. On closer inspection, the regular coordination geometry of the complex consists of octahedral NiCl6 in the center linked by two symmetrically bridging chloride ions to square-pyramidal NiCl5 on each end. A long semicoordinate bond is formed by each of the terminal Ni(II) cations, to give a 5+1 coordination geometry and form an asymmetric bridge to the central Ni(II) cation. The dimethylammonium cations surround the complex with an extensive hydrogen-bonding network, linking the complex to the noncoordinated chloride ions. Asymmetric bridging in the complex arises from short hydrogen bonds from the same dimethylammonium cation to the apical and asymmetric bridging chloride ions, causing the complex to scissor outward. PMID- 19805877 TI - New organically templated vanadium tellurites: (H2pn)[V2TeO8] (pn is propane-1,3 diamine). AB - The title compound, poly[propane-1,3-diaminium hexa-mu-oxido dioxidotellurium(IV)divanadium(V)], (C3H12N2)[V2O8Te] or (H2pn)[V2TeO8] (pn is propane-1,3-diamine), contains a two-dimensional anionic layer and the diprotonated pn cation for charge compensation. The anionic layer consists of pyrovanadates and [TeO3] pyramids, which are linked alternately through corner sharing to form a one-dimensional chain. These one-dimensional chains are crosslinked through two weak Te-O bonds, constructing an anionic layer. Hydrogen bonds are observed involving the diprotonated pn cation and the O atoms of the anionic framework. PMID- 19805876 TI - A three-dimensional supramolecular vanadium hydroxylamide complex: poly[di-mu2 aqua-bis(hydroxylamido)-mu3-malonato-oxidosodiumvanadium(V)]. AB - The crystal structure of the title compound, [NaV(C3H2O4)(NH2O)2O(H2O)2], is built up of NaO6 and VO5N2 polyhedra connected through malonate bridges. The NaO6 octahedra are linked by edge sharing in the equatorial plane to form one dimensional infinite chains. These chains are linked together by the malonate bridges to form two-dimensional layers. The distorted VO5N2 pentagonal bipyramid is grafted on to the layer by a malonate carboxylate O atom. Adjacent layers are connected through O-H...O and N-H...O hydrogen bonds to build up a three dimensional supramolecular structure. PMID- 19805878 TI - A stair-like two-dimensional silver(I) coordination polymer of N'-(3 cyanobenzylidene)nicotinohydrazide. AB - The structure of the title compound, poly[[[mu3-N'-(3 cyanobenzylidene)nicotinohydrazide]silver(I)] hexafluoroarsenate], {[Ag(C14H10N4O)](AsF6)}n, at 173 K exhibits a novel stair-like two-dimensional layer and a three-dimensional supramolecular framework through C-H...Ag hydrogen bonds. The Ag(I) cation is coordinated by three N atoms and one O atom from N'-(3 cyanobenzylidene)nicotinohydrazide (L) ligands, resulting in a distorted tetrahedral coordination geometry. The organic ligand acts as a mu3-bridging ligand through the pyridyl and carbonitrile N atoms and deviates from planarity in order to adapt to the coordination geometry. Two ligands bridge two Ag(I) cations to construct a small 2+2 Ag2L2 ring. Four ligands bridge one Ag(I) cation from each of four of these small rings to form a large grid. An interesting stair like two-dimensional (3,6)-net is formed through Ag(I) metal centres acting as three-connection nodes and through L molecules as tri-linkage spacers. PMID- 19805879 TI - 3-[(E)-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)iminomethyl]quinolin-2(1H)-one: chains built by pi-stacking of hydrogen-bonded R(2)(2)(8) dimers. AB - In the title compound, C23H22N4O, there is evidence for some bond fixation in the aryl component of the quinolinone unit. Pairs of molecules related by inversion are linked into R(2)(2)(8) dimers by almost linear N-H...O hydrogen bonds, and dimers related by inversion are linked into chains by a single aromatic pi-pi stacking interaction. PMID- 19805880 TI - Rotationally disordered phase of 1,3-dibromo-5-iodo-2,4,6-trimethylbenzene at 293 K. AB - In the crystal state at room temperature, the molecule of dibromoiodomesitylene (1,3-dibromo-5-iodo-2,4,6-trimethylbenzene), C9H9Br2I, is prone to strong disorder, apparently involving only the three halogen sites (occupied identically by 66.7% Br and 33.3% I). This disorder, of the rotational type according to previously published NMR measurements, corresponds to fast 2pi/3 stochastic in plane reorientations of the whole molecule between three discernable locations. This kind of rotational disorder can be revealed for the first time by diffractometry thanks to the C2v idealized molecular symmetry of the title compound, although it has been indirectly suspected at room temperature in other trihalogenomesitylenes of similar crystal packing but of D3h molecular symmetry. The average endocyclic angles facing the Br/I sites and the methyl groups are 124.14 (6) and 115.85 (2) degrees, respectively. The angle between the normal to the aromatic ring and the normal to the (100) plane is 4.1 degrees. TLS analysis indicates that only the aromatic ring and the methyl groups behave as a rigid body with respect to the thermal librations. PMID- 19805881 TI - Lenalidomide, an antineoplastic drug, and its hemihydrate. AB - The crystal structures of lenalidomide [systematic name: (RS)-3-(4-amino-1 oxoisoindolin-2-yl)piperidine-2,6-dione], C13H13N3O3, (I), an antineoplastic drug, and its hemihydrate, C13H13N3O3.0.5H2O, (II), have been determined by single-crystal X-ray diffraction analysis. The overall conformation of the molecule defined by the orientation of the two ring portions, viz. pyridinedione and isoindolinone, is twisted in both structures. The influence of the self complementary pyridinedione ring is seen in the crystal packing of both structures through its involvement in forming hydrogen-bonded dimers, although alternate dione O atoms are utilized. An extensive series of N-H...O hydrogen bonds link the dimers into two-dimensional supramolecular arrays built up from infinite chains. The water molecule in (II) has a cohesive function, connecting three lenalidomide molecules by hydrogen bonds. The significance of this study lies in the analysis of the interactions in these structures and the aggregations occurring via hydrogen bonds in the hydrated and dehydrated crystalline forms of the title compound. PMID- 19805882 TI - 3-(1H-pyrrol-2-yl)-1H-pyrazole forms an unusual hydrogen-bonded two-dimensional (3,4)-connected net. AB - The title compound, C7H7N3, is the first crystallographically characterized 1H pyrrolyl-1H-pyrazole derivative and contains two unique molecules in its asymmetric unit (Z' = 2). These molecules associate into centrosymmetric tetramers through N-H...N hydrogen bonding, including a cyclic dimerization of one of the two unique pyrazole rings. These tetramers are linked further by two weaker N-H...pi contacts to give a novel two-dimensional (3,4)-connected net with a (3(2).8)(2)(3.8(2))(2) topology. PMID- 19805883 TI - Coulombic interactions, hydrogen bonding and supramolecular chirality in pyridinium trifluoromethanesulfonate. AB - The title compound, C5H6N+.CF3SO3-, was serendipitously crystallized in the chiral space group P4(3)2(1)2. The component entities associate into hydrogen bonded helical chains, which propagate along the a and b axes of the crystal, with an alternating disposition of the cations and anions along the chain. N H...O charge-assisted hydrogen bonds, from each pyridinium cation to two adjacent trifluoromethanesulfonate anions and from every anion to two different cations, direct the formation of the supramolecular chiral arrays. The crystal packing exhibits nonconventional C-H...O and C-H...F hydrogen bonds between the components. The observed structure demonstrates induction of supramolecular chirality by a combination of Coulombic attractions and intermolecular hydrogen bonds. PMID- 19805884 TI - Three quinolone compounds featuring O...I halogen bonding. AB - Ethyl 1-ethyl-6-iodo-4-oxo-1,4-dihydroquinoline-3-carboxylate, C14H14INO3, (I), and ethyl 1-cyclopropyl-6-iodo-4-oxo-1,4-dihydroquinoline-3-carboxylate, C15H14INO3, (II), have isomorphous crystal structures, while ethyl 1 dimethylamino-6-iodo-4-oxo-1,4-dihydroquinoline-3-carboxylate, C14H15IN2O3, (III), possesses a different solid-state supramolecular architecture. In all three structures, O...I halogen-bonding interactions connect the quinolone molecules into infinite chains parallel to the unique crystallographic b axis. In (I) and (II), these molecular chains are arranged in (101) layers, via pi-pi stacking and C-H...pi interactions, and these layers are then interlinked by C H...O interactions. The structural fragments involved in the C-H...O interactions differ between (I) and (II), accounting for the observed difference in planarity of the quinolone moieties in the two isomorphous structures. In (III), C-H...O and C-H...pi interactions form (100) molecular layers, which are crosslinked by O...I and C-H...I interactions. PMID- 19805885 TI - (E)-2-[(4-chlorophenyl)iminomethyl]-5-methoxyphenol and (E)-2-[(2 chlorophenyl)iminomethyl]-5-methoxyphenol: X-ray and DFT-calculated structures. AB - The crystal structures of the title 4-chlorophenyl, (I), and 2-chlorophenyl, (II), compounds, both C14H12ClNO2, have been determined using X-ray diffraction techniques and the molecular structures have also been optimized at the B3LYP/6 31 G(d,p) level using density functional theory (DFT). The X-ray study shows that the title compounds both have strong intramolecular O-H...N hydrogen bonds and that the crystal networks are primarily determined by weak C-H...pi and van der Waals interactions. The strong intramolecular O-H...N hydrogen bond is evidence of the preference for the phenol-imine tautomeric form in the solid state. The IR spectra of the compounds were recorded experimentally and also calculated for comparison. The results from both the experiment and theoretical calculations are compared in this study. PMID- 19805886 TI - Pseudomerohedrally twinned monoclinic structure of unfolded 'free' nonactin: comparative analysis of its large conformational change upon encapsulation of alkali metal ions. AB - The title compound, C40H64O12, crystallizes in a pseudomerohedrally twinned primitive monoclinic cell with similar contributions of the two twin components. There are two symmetry-independent half-molecules of nonactin in the asymmetric unit. Each molecule has a pseudo-S4 symmetry and resides on a crystallographic twofold axis; the axes pass through the molecular center of mass and are perpendicular to the plane of the macrocycle. The literature description of the room-temperature structure of nonactin as an order-disorder structure in an orthorhombic unit cell is corrected. We report a low-temperature high-precision ordered structure of ;free' nonactin that allowed for the first time precise determination of its bond distances and angles. It possesses an unfolded and more planar geometry than its complexes with encapsulated Na+, K+, Cs+, Ca2+ or NH4+ cations that exhibit more isometric overall conformations. PMID- 19805887 TI - Hydrogen-bond-directed supramolecular arrays in 4,4'-bipyridinium tetrachloroterephthalate dihydrate and bis(1,10-phenanthrolinium) tetrachloroterephthalate tetrachloroterephthalic acid trihydrate. AB - The title compounds, C10H10N(2)(2+).C8Cl4O(4)(2-).2H2O, (I), and 2C12H9N2+.C8Cl4O(4)(2-).C8H2Cl4O4.3H2O, (II), both crystallize as charge-transfer organic salts with the dianionic or neutral acid components lying on inversion centres. The acid and base subunits in (I) arrange alternately to generate a linear tape motif via N-H...O hydrogen bonds; these tapes are further combined into a three-dimensional architecture through multiple O-H...O and C-H...O interactions involving solvent water molecules. In contrast, the neutral and anionic acid components in (II) are linked to form a zigzag chain by means of O H...O hydrogen bonds between acid groups, with dangling 1,10-phenanthrolinium units connected to these chains by carboxylate-pyridinium interactions with R(2)(2)(7) hydrogen-bond notation. Adjacent chains are further extended to result in a two-dimensional corrugated layer network via pi-pi interactions. Inter-ion Cl...O interactions are also found in both (I) and (II). PMID- 19805888 TI - Channel-forming solvates of 6-chloro-2,5-dihydroxypyridine and its solvent-free tautomer 6-chloro-5-hydroxy-2-pyridone. AB - On crystallization from CHCl3, CCl4, CH2ClCH2Cl and CHCl2CHCl2, 6-chloro-5 hydroxy-2-pyridone, C5H4ClNO2, (I), undergoes a tautomeric rearrangement to 6 chloro-2,5-dihydroxypyridine, (II). The resulting crystals, viz. 6-chloro-2,5 dihydroxypyridine chloroform 0.125-solvate, C5H4ClNO(2).0.125CHCl3, (IIa), 6 chloro-2,5-dihydroxypyridine carbon tetrachloride 0.125-solvate, C5H4ClNO(2)..0.125CCl4, (IIb), 6-chloro-2,5-dihydroxypyridine 1,2-dichloroethane solvate, C5H4ClNO2.C2H4Cl2, (IIc), and 6-chloro-2,5-dihydroxypyridine 1,1,2,2 tetrachloroethane solvate, C5H4ClNO2.C2H2Cl4, (IId), have I4(1)/a symmetry, and incorporate extensively disordered solvent in channels that run the length of the c axis. Upon gentle heating to 378 K in vacuo, these crystals sublime to form solvent-free crystals with P2(1)/n symmetry that are exclusively the pyridone tautomer, (I). In these sublimed pyridone crystals, inversion-related molecules form R(2)(2)(8) dimers via pairs of N-H...O hydrogen bonds. The dimers are linked by O-H...O hydrogen bonds into R(4)(6)(28) motifs, which join to form pleated sheets that stack along the a axis. In the channel-containing pyridine solvate crystals, viz. (IIa)-(IId), two independent host molecules form an R(2)(2)(8) dimer via a pair of O-H...N hydrogen bonds. One molecule is further linked by O H...O hydrogen bonds to two 4(1) screw-related equivalents to form a helical motif parallel to the c axis. The other independent molecule is O-H...O hydrogen bonded to two 4 related equivalents to form tetrameric R(4)(4)(28) rings. The dimers are pi-pi stacked with inversion-related dimers, which in turn stack the R(4)(4)(28) rings along c to form continuous solvent-accessible channels. CHCl3, CCl4, CH2ClCH2Cl and CHCl2CHCl2 solvent molecules are able to occupy these channels but are disordered by virtue of the 4 site symmetry within the channels. PMID- 19805889 TI - Hydrogen-bonded structures of the isomeric compounds of quinoline with 2-chloro-5 nitrobenzoic acid, 3-chloro-2-nitrobenzoic acid, 4-chloro-2-nitrobenzoic acid and 5-chloro-2-nitrobenzoic acid. AB - The structures of four isomeric compounds, all C7H4ClNO4.C9H7N, of quinoline with chloro- and nitro-substituted benzoic acid, namely, 2-chloro-5-nitrobenzoic acid quinoline (1/1), (I), 3-chloro-2-nitrobenzoic acid-quinoline (1/1), (II), 4 chloro-2-nitrobenzoic acid-quinoline (1/1), (III), and 5-chloro-2-nitrobenzoic acid-quinoline (1/1), (IV), have been determined at 185 K. In each compound, a short hydrogen bond is observed between the pyridine N atom and a carboxyl O atom. The N...O distances are 2.6476 (13), 2.5610 (13), 2.5569 (12) and 2.5429 (12) A for (I), (II), (III) and (IV), respectively. Although in (I) the H atom in the hydrogen bond is located at the O site, in (II), (III) and (IV) the H atom is disordered in the hydrogen bond over two positions with (N site):(O site) occupancies of 0.39 (3):0.61 (3), 0.47 (3):0.53 (3) and 0.65 (3):0.35 (3), respectively. PMID- 19805890 TI - A new polymorph of 2-methyl-6-nitroaniline. AB - A new crystal form of 2-methyl-6-nitroaniline, C7H8N2O2, crystallizing with Z' = 2 in the space group P2(1)/c, has been identified during screening for salts and cocrystals. The different N-H...O hydrogen-bonding synthons result in linear V shaped chains in the new polymorph, rather than the helical chain arrangement seen in the known form where Z' = 1. The presence of a second component during crystallization appears to have determined the resultant crystal form of 2-methyl 6-nitroaniline. PMID- 19805891 TI - Proton transfer versus nontransfer in compounds of the diazo-dye precursor 4 (phenyldiazenyl)aniline (aniline yellow) with strong organic acids: the 5 sulfosalicylate and the dichroic benzenesulfonate salts, and the 1:2 adduct with 3,5-dinitrobenzoic acid. AB - The structures of two 1:1 proton-transfer red-black dye compounds formed by reaction of aniline yellow [4-(phenyldiazenyl)aniline] with 5-sulfosalicylic acid and benzenesulfonic acid, and a 1:2 nontransfer adduct compound with 3,5 dinitrobenzoic acid have been determined at either 130 or 200 K. The compounds are 2-(4-aminophenyl)-1-phenylhydrazin-1-ium 3-carboxy-4-hydroxybenzenesulfonate methanol solvate, C12H12N3+.C7H5O6S-.CH3OH, (I), 2-(4-aminophenyl)-1 phenylhydrazin-1-ium 4-(phenyldiazenyl)anilinium bis(benzenesulfonate), 2C12H12N3+.2C6H5O3S-, (II), and 4-(phenyldiazenyl)aniline-3,5-dinitrobenzoic acid (1/2), C12H11N3.2C7H4N2O6, (III). In compound (I), the diazenyl rather than the aniline group of aniline yellow is protonated, and this group subsequently takes part in a primary hydrogen-bonding interaction with a sulfonate O-atom acceptor, producing overall a three-dimensional framework structure. A feature of the hydrogen bonding in (I) is a peripheral edge-on cation-anion association also involving aromatic C-H...O hydrogen bonds, giving a conjoint R(1)(2)(6)R(1)(2)(7)R(2)(1)(4) motif. In the dichroic crystals of (II), one of the two aniline yellow species in the asymmetric unit is diazenyl-group protonated, while in the other the aniline group is protonated. Both of these groups form hydrogen bonds with sulfonate O-atom acceptors and these, together with other associations, give a one-dimensional chain structure. In compound (III), rather than proton transfer, there is preferential formation of a classic R(2)(2)(8) cyclic head-to-head hydrogen-bonded carboxylic acid homodimer between the two 3,5-dinitrobenzoic acid molecules, which, in association with the aniline yellow molecule that is disordered across a crystallographic inversion centre, results in an overall two-dimensional ribbon structure. This work has shown the correlation between structure and observed colour in crystalline aniline yellow compounds, illustrated graphically in the dichroic benzenesulfonate compound. PMID- 19805892 TI - M K Chandrashekaran (1937-2009). PMID- 19805893 TI - Branches in the plant world. PMID- 19805894 TI - Estradiol can modulate sensory processing with rapid and longer term consequences. PMID- 19805896 TI - R A Fisher, design theory, and the Indian connection. AB - Design Theory, a branch of mathematics, was born out of the experimental statistics research of the population geneticist R A Fisher and of Indian mathematical statisticians in the 1930s. The field combines elements of combinatorics, finite projective geometries, Latin squares, and a variety of further mathematical structures, brought together in surprising ways. This essay will present these structures and ideas as well as how the field came together, in itself an interesting story. PMID- 19805895 TI - Of pungency, pain, and naked mole rats: chili peppers revisited. PMID- 19805897 TI - B P Belousov and his reaction. AB - The generation to which Boris Pavlovich Belousov (1893-1976) belonged has almost disappeared. The archives hold only a few documents about his life and basically secret research. This article is a brief biography of Belousov and attempts to reconstruct what lay behind his famous discovery of the oscillatory homogeneous chemical reaction named after him. PMID- 19805898 TI - What history tells us XVIII. When functional biologists propose mechanisms of evolution. PMID- 19805899 TI - Thermal stability of alpha-amylase in aqueous cosolvent systems. AB - The activity and thermal stability of alpha-amylase were studied in the presence of different concentrations of trehalose, sorbitol, sucrose and glycerol. The optimum temperature of the enzyme was found to be 50 +/- 2 degrees C. Further increase in temperature resulted in irreversible thermal inactivation of the enzyme. In the presence of cosolvents, the rate of thermal inactivation was found to be significantly reduced. The apparent thermal denaturation temperature (Tm) app and activation energy (Ea) of alpha-amylase were found to be significantly increased in the presence of cosolvents in a concentration-dependent manner. In the presence of 40% trehalose, sorbitol, sucrose and glycerol, increments in the (Tm)app were 20 degrees C, 14 degrees C, 13 degrees C and 9 degrees C, respectively. The Ea of thermal denaturation of alpha-amylase in the presence of 20% (w/v) trehalose, sorbitol, sucrose and glycerol was found to be 126, 95, 90 and 43 kcal/mol compared with a control value of 40 kcal/mol. Intrinsic and 8 anilinonaphathalene-1-sulphonic acid (ANS) fluorescence studies indicated that thermal denaturation of the enzyme was accompanied by exposure of the hydrophobic cluster on the protein surface. Preferential interaction parameters indicated extensive hydration of the enzyme in the presence of cosolvents. PMID- 19805900 TI - GalNAc-T14 may be involved in regulating the apoptotic action of IGFBP-3. AB - Insulin-like growth factor binding protein-3 (IGFBP-3) is known to induce apoptosis in an insulin-like growth factor (IGF)-dependent and IGF-independent manner, but the mechanism underlying the IGF-independent effects remains unclear. Polypeptide N-acetylgalactosaminyltransferase 14 (GalNAc-T14) is a novel IGFBP-3 binding partner. In this paper, small interference RNA (siRNA) targeting GalNAc T14 was used to examine whether GalNAc-T14 affects the apoptotic action of IGFBP 3. Using semi-quantitative reverse-transcriptase polymerase chain reaction (RT PCR) and western blot analysis, we determined that GalNAc-T14 expression was downregulated by the siRNA directed against GalNAc-T14. Apoptosis analysis of IGFBP-3-overexpressing cells treated with siRNA against GalNAc-T14 was performed to determine if GalNAc-T14 was specifically involved in IGFBP-3 signalling. The results, as determined by flow cytometric analysis and caspase-3 assay, showed that the extent of apoptosis induced by IGFBP-increased with RNA interference (RNAi) knockdown of GalNAc-T14. Our data suggest that GalNAc-T14 influences the apoptotic action of IGFBP-3 and might mediate the signalling pathway of IGFBP-3. Experiments to determine the role of GalNAc-T14 in the regulation of apoptosis induced by IGFBP-3 are under way. PMID- 19805901 TI - Single-nucleotide variations associated with Mycobacterium tuberculosis KwaZulu Natal strains. AB - The occurrence of drug resistance in Mycobacterium tuberculosis, the aetiological agent of tuberculosis (TB), is hampering the management and control of TB in the world. Here we present a computational analysis of recently sequenced drug sensitive (DS), multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains of M. tuberculosis. Single-nucleotide variations (SNVs) were identified in a pair-wise manner using the anchor-based whole genome comparison (ABWGC) tool and its modified version. For this analysis, four fully sequenced genomes of different strains of M. tuberculosis were taken along with three KwaZulu-Natal (KZN) strains isolated from South Africa including one XDR and one MDR strain. KZN strains were compared with other fully sequenced strains and also among each other. The variations were analysed with respect to their biological influence as a result of either altered structure or synthesis. The results suggest that the DR phenotype may be due to changes in a number of genes. The database on KZN strains can be accessed through the website http://mirna.jnu.ac.in/mgdd/. PMID- 19805902 TI - Single-nucleotide polymorphisms in peroxisome proliferator-activated receptor gamma and their association with plasma levels of resistin and the metabolic syndrome in a South Indian population. AB - Studies on the association of the Pro12Ala and C1431T polymorphisms of PPAR? with diabetes and obesity have revealed extensive population-dependent variations. However, association of these polymorphisms with the metabolic syndrome and its individual components has not been well investigated in the Indian population. The Indian population harbours the maximum number of diabetics in the world who are thus more susceptible to metabolic disorders. We screened a South Indian population (N=699) for a possible association of these polymorphisms with the metabolic syndrome (MS) and type 2 diabetes. We also investigated the correlation of these two single-nucleotide polymorphisms (SNPs) with plasma resistin levels. The C1431T SNP was associated with higher levels of plasma resistin (P=0.017). Furthermore, C1431T was associated with resistin in different tertiles. Prevalence of the 'Pro-C' haplotype decreased with increasing tertiles of resistin (84.1% to 75.4%, P=0.037). Plasma resistin levels were not found to be associated with MS and type 2 diabetes. These results point to a likely association of plasma resistin levels with PPAR? polymorphisms in the Indian population. PMID- 19805903 TI - BRCA1 and BRCA2 germline mutation analysis among Indian women from south India: identification of four novel mutations and high-frequency occurrence of 185delAG mutation. AB - Mutations in the BRCA1 and BRCA2 genes profoundly increase the risk of developing breast and/or ovarian cancer among women. To explore the contribution of BRCA1 and BRCA2 mutations in the development of hereditary breast cancer among Indian women, we carried out mutation analysis of the BRCA1 and BRCA2 genes in 61 breast or ovarian cancer patients from south India with a positive family history of breast and/or ovarian cancer. Mutation analysis was carried out using conformation-sensitive gel electrophoresis (CSGE) followed by sequencing. Mutations were identified in 17 patients (28.0%); 15 (24.6%) had BRCA1 mutations and two (3.28%) had BRCA2 mutations. While no specific association between BRCA1 or BRCA2 mutations with cancer type was seen, mutations were more often seen in families with ovarian cancer. While 40% (4/10) and 30.8% (4/12) of families with ovarian or breast and ovarian cancer had mutations, only 23.1% (9/39) of families with breast cancer carried mutations in the BRCA1 and BRCA2 genes. In addition, while BRCA1 mutations were found in all age groups, BRCA2 mutations were found only in the age group of < or =40 years. Of the BRCA1 mutations, there were three novel mutations (295delCA; 4213T-->A; 5267T-->G) and three mutations that have been reported earlier. Interestingly, 185delAG, a BRCA1 mutation which occurs at a very high frequency in Ashkenazi Jews, was found at a frequency of 16.4% (10/61). There was one novel mutation (4866insT) and one reported mutation in BRCA2. Thus, our study emphasizes the importance of mutation screening in familial breast and/or ovarian cancers, and the potential implications of these findings in genetic counselling and preventive therapy. PMID- 19805904 TI - A simple and efficient Agrobacterium-mediated procedure for transformation of tomato. AB - We describe a highly efficient and reproducible Agrobacterium-mediated transformation protocol applicable to several varieties of tomato (Solanum lycopersicum, earlier known as Lycopersicum esculentum). Conditions such as co cultivation period, bacterial concentration, concentration of benzyl amino purine (BAP), zeatin and indole acetic acid (IAA) were optimized. Co-cultivation of explants with a bacterial concentration of 108 cells/ml for three days on 2 mg/l BAP, followed by regeneration on a medium containing 1 mg/ml zeatin resulted in a transformation frequency of 41.4%. Transformation of tomato plants was confirmed by Southern blot analysis and beta-glucuronidase (GUS) assay. The protocol developed showed very high efficiency of transformation for tomato varieties Pusa Ruby, Arka Vikas and Sioux. The optimized transformation procedure is simple, efficient and does not require tobacco, Petunia, tomato suspension feeder layer or acetosyringone. PMID- 19805905 TI - Association preference and mechanism of kin recognition in tadpoles of the toad Bufo melanostictus. AB - In experiments with specially designed choice tanks, tadpoles of Bufo melanostictus spend significantly greater amounts of time near kin than near non kin. However, in the absence of kin members, they prefer to spend more time near non-kin rather than stay away in isolation in the opposite blank zone with no company. This implies that association of toad tadpoles with their kin is due to attraction rather than repulsion from non-kin. Experiments designed to elucidate the sensory basis of kin recognition showed that toad tadpoles recognize their kin based on chemical cues rather than visual cues. They can also discriminate between homospecific non-kin and heterospecific (Sphaerotheca breviceps) tadpoles since the tadpoles spent significantly greater amounts of time near the former than near the latter. These findings suggest that where kin members are unavailable, selection may have favoured living with non-kin so as to derive benefits from group living and that a phenotype-matching mechanism may operate for both kin and species discrimination in B. melanostictus. PMID- 19805906 TI - Quantitative effect and regulatory function of cyclic adenosine 5'-phosphate in Escherichia coli. AB - Cyclic adenosine 5'-phosphate (cAMP) is a global regulator of gene expression in Escherichia coli. Despite decades of intensive study, the quantitative effect and regulatory function of cAMP remain the subjects of considerable debate. Here, we analyse the data in the literature to show that: (a) In carbon-limited cultures (including cultures limited by glucose), cAMP is at near-saturation levels with respect to expression of several catabolic promoters (including lac, ara and gal). It follows that cAMP receptor protein (CRP) cAMP-mediated regulation cannot account for the strong repression of these operons in the presence of glucose. (b) The cAMP levels in carbon-excess cultures are substantially lower than those observed in carbon-limited cultures under these conditions, the expression of catabolic promoters is very sensitive to variation of cAMP levels. (c)=CRPcAMP invariably activates the expression of catabolic promoters, but it appears to inhibit the expression of anabolic promoters. (d) These results suggest that the physiological function of cAMP is to maintain homeostatic energy levels. In carbon-limited cultures, growth is limited by the supply of energy; the cAMP levels therefore increase to enhance energy accumulation by activating the catabolic promoters and inhibiting the anabolic promoters. Conversely, in carbonexcess cultures, characterized by the availability of excess energy, the cAMP levels decrease in order to depress energy accumulation by inhibiting the catabolic promoters and activating the anabolic promoters. PMID- 19805907 TI - Structure-function-folding relationships and native energy landscape of dynein light chain protein: nuclear magnetic resonance insights. AB - The detailed characterization of the structure, dynamics and folding process of a protein is crucial for understanding the biological functions it performs. Modern biophysical and nuclear magnetic resonance (NMR) techniques have provided a way to obtain accurate structural and thermodynamic information on various species populated on the energy landscape of a given protein. In this context, we review here the structure-function-folding relationship of an important protein, namely, dynein light chain protein (DLC8). DLC8, the smallest subunit of the dynein motor complex, acts as a cargo adaptor. The protein exists as a dimer under physiological conditions and dissociates into a pure monomer below pH 4. Cargo binding occurs at the dimer interface. Dimer stability and relay of perturbations through the dimer interface are anticipated to be playing crucial roles in the variety of functions the protein performs. NMR investigations have provided great insights into these aspects of DLC8 in recent years. PMID- 19805908 TI - Influenza A (H1N1) 2009: a pandemic alarm. AB - At this critical juncture when the world has not yet recovered from the threat of avian influenza, the virus has returned in the disguise of swine influenza, a lesser known illness common in pigs. It has reached pandemic proportions in a short time span with health personnel still devising ways to identify the novel H1N1 virus and develop vaccines against it. The H1N1 virus has caused a considerable number of deaths within the short duration since its emergence. Presently, there are no effective methods to contain this newly emerged virus. Therefore, a proper and clear insight is urgently required to prevent an outbreak in the future and make preparations that may be planned well in advance. This review is an attempt to discuss the historical perspective of the swine flu virus, its epidemiology and route of transmission to better understand the various control measures that may be taken to fight the danger of a global pandemic. PMID- 19805909 TI - Small-molecule screening using a human primary cell model of HIV latency identifies compounds that reverse latency without cellular activation. AB - The development of highly active antiretroviral therapy (HAART) to treat individuals infected with HIV-1 has dramatically improved patient outcomes, but HAART still fails to cure the infection. The latent viral reservoir in resting CD4+ T cells is a major barrier to virus eradication. Elimination of this reservoir requires reactivation of the latent virus. However, strategies for reactivating HIV-1 through nonspecific T cell activation have clinically unacceptable toxicities. We describe here the development of what we believe to be a novel in vitro model of HIV-1 latency that we used to search for compounds that can reverse latency. Human primary CD4+ T cells were transduced with the prosurvival molecule Bcl-2, and the resulting cells were shown to recapitulate the quiescent state of resting CD4+ T cells in vivo. Using this model system, we screened small-molecule libraries and identified a compound that reactivated latent HIV-1 without inducing global T cell activation, 5-hydroxynaphthalene-1,4 dione (5HN). Unlike previously described latency-reversing agents, 5HN activated latent HIV-1 through ROS and NF-kappaB without affecting nuclear factor of activated T cells (NFAT) and PKC, demonstrating that TCR pathways can be dissected and utilized to purge latent virus. Our study expands the number of classes of latency-reversing therapeutics and demonstrates the utility of this in vitro model for finding strategies to eradicate HIV-1 infection. PMID- 19805910 TI - Adoptive immunotherapy with liver allograft-derived lymphocytes induces anti-HCV activity after liver transplantation in humans and humanized mice. AB - After liver transplantation in HCV-infected patients, the virus load inevitably exceeds pre-transplantation levels. This phenomenon reflects suppression of the host-effector immune responses that control HCV replication by the immunosuppressive drugs used to prevent rejection of the transplanted liver. Here, we describe an adoptive immunotherapy approach, using lymphocytes extracted from liver allograft perfusate (termed herein liver allograft-derived lymphocytes), which includes an abundance of NK/NKT cells that mounted an anti HCV response in HCV-infected liver transplantation recipients, despite the immunosuppressive environment. This therapy involved intravenously injecting patients 3 days after liver transplantation with liver allograft-derived lymphocytes treated with IL-2 and the CD3-specific mAb OKT3. During the first month after liver transplantation, the HCV RNA titers in the sera of recipients who received immunotherapy were markedly lower than those in the sera of recipients who did not receive immunotherapy. We further explored these observations in human hepatocyte-chimeric mice, in which mouse hepatocytes were replaced by human hepatocytes. These mice unfailingly developed HCV infections after inoculation with HCV-infected human serum. However, injection of human liver-derived lymphocytes treated with IL-2/OKT3 completely prevented HCV infection. Furthermore, an in vitro study using genomic HCV replicon-containing hepatic cells revealed that IFN-gamma-secreting cells played a pivotal role in such anti-HCV responses. Thus, our study presents what we believe to be a novel paradigm for the inhibition of HCV replication in HCV-infected liver transplantation recipients. PMID- 19805911 TI - Impaired autophagic flux mediates acinar cell vacuole formation and trypsinogen activation in rodent models of acute pancreatitis. AB - The pathogenic mechanisms underlying acute pancreatitis are not clear. Two key pathologic acinar cell responses of this disease are vacuole accumulation and trypsinogen activation. We show here that both result from defective autophagy, by comparing the autophagic responses in rodent models of acute pancreatitis to physiologic autophagy triggered by fasting. Pancreatitis-induced vacuoles in acinar cells were greater in number and much larger than those induced with fasting. Degradation of long-lived proteins, a measure of autophagic efficiency, was markedly inhibited in in vitro pancreatitis, while it was stimulated by acinar cell starvation. Further, processing of the lysosomal proteases cathepsin L (CatL) and CatB into their fully active, mature forms was reduced in pancreatitis, as were their activities in the lysosome-enriched subcellular fraction. These findings indicate that autophagy is retarded in pancreatitis due to deficient lysosomal degradation caused by impaired cathepsin processing. Trypsinogen activation occurred in pancreatitis but not with fasting and was prevented by inhibiting autophagy. A marker of trypsinogen activation partially localized to autophagic vacuoles, and pharmacologic inhibition of CatL increased the amount of active trypsin in acinar cells. The results suggest that retarded autophagy is associated with an imbalance between CatL, which degrades trypsinogen and trypsin, and CatB, which converts trypsinogen into trypsin, resulting in intra-acinar accumulation of active trypsin in pancreatitis. Thus, deficient lysosomal degradation may be a dominant mechanism for increased intra acinar trypsin in pancreatitis. PMID- 19805912 TI - Suppression of KATP channel activity protects murine pancreatic beta cells against oxidative stress. AB - The enhanced oxidative stress associated with type 2 diabetes mellitus contributes to disease pathogenesis. We previously identified plasma membrane associated ATP-sensitive K+ (KATP) channels of pancreatic beta cells as targets for oxidants. Here, we examined the effects of genetic and pharmacologic ablation of KATP channels on loss of mouse beta cell function and viability following oxidative stress. Using mice lacking the sulfonylurea receptor type 1 (Sur1) subunit of KATP channels, we found that, compared with insulin secretion by WT islets, insulin secretion by Sur1-/- islets was less susceptible to oxidative stress induced by the oxidant H2O2. This was likely, at least in part, a result of the reduced ability of H2O2 to hyperpolarize plasma membrane potential and reduce cytosolic free Ca2+ concentration ([Ca2+]c) in the Sur1-/- beta cells. Remarkably, Sur1-/- beta cells were less prone to apoptosis induced by H2O2 or an NO donor than WT beta cells, despite an enhanced basal rate of apoptosis. This protective effect was attributed to upregulation of the antioxidant enzymes SOD, glutathione peroxidase, and catalase. Upregulation of antioxidant enzymes and reduced sensitivity of Sur1-/- cells to H2O2-induced apoptosis were mimicked by treatment with the sulfonylureas tolbutamide and gliclazide. Enzyme upregulation and protection against oxidant-induced apoptosis were abrogated by agents lowering [Ca2+]c. Sur1-/- mice were less susceptible than WT mice to streptozotocin-induced beta cell destruction and subsequent hyperglycemia and death, which suggests that loss of KATP channel activity may protect against streptozotocin-induced diabetes in vivo. PMID- 19805914 TI - Real-time imaging of de novo arteriovenous malformation in a mouse model of hereditary hemorrhagic telangiectasia. AB - Arteriovenous malformations (AVMs) are vascular anomalies where arteries and veins are directly connected through a complex, tangled web of abnormal arteries and veins instead of a normal capillary network. AVMs in the brain, lung, and visceral organs, including the liver and gastrointestinal tract, result in considerable morbidity and mortality. AVMs are the underlying cause of three major clinical symptoms of a genetic vascular dysplasia termed hereditary hemorrhagic telangiectasia (HHT), which is characterized by recurrent nosebleeds, mucocutaneous telangiectases, and visceral AVMs and caused by mutations in one of several genes, including activin receptor-like kinase 1 (ALK1). It remains unknown why and how selective blood vessels form AVMs, and there have been technical limitations to observing the initial stages of AVM formation. Here we present in vivo evidence that physiological or environmental factors such as wounds in addition to the genetic ablation are required for Alk1-deficient vessels to develop to AVMs in adult mice. Using the dorsal skinfold window chamber system, we have demonstrated for what we believe to be the first time the entire course of AVM formation in subdermal blood vessels by using intravital bright-field images, hyperspectral imaging, fluorescence recordings of direct arterial flow through the AV shunts, and vascular casting techniques. We believe our data provide novel insights into the pathogenetic mechanisms of HHT and potential therapeutic approaches. PMID- 19805913 TI - Syndecan-1 is the primary heparan sulfate proteoglycan mediating hepatic clearance of triglyceride-rich lipoproteins in mice. AB - Elevated plasma triglyceride levels represent a risk factor for premature atherosclerosis. In mice, accumulation of triglyceride-rich lipoproteins can occur if sulfation of heparan sulfate in hepatocytes is diminished, as this alters hepatic lipoprotein clearance via heparan sulfate proteoglycans (HSPGs). However, the relevant HSPG has not been determined. In this study, we found by RT PCR analysis that mouse hepatocytes expressed the membrane proteoglycans syndecan 1, -2, and -4 and glypican-1 and -4. Analysis of available proteoglycan-deficient mice showed that only syndecan-1 mutants (Sdc1-/- mice) accumulated plasma triglycerides. Sdc1-/- mice also exhibited prolonged circulation of injected human VLDL and intestinally derived chylomicrons. We found that mice lacking both syndecan-1 and hepatocyte heparan sulfate did not display accentuated triglyceride accumulation compared with single mutants, suggesting that syndecan 1 is the primary HSPG mediating hepatic triglyceride clearance. Immunoelectron microscopy showed that syndecan-1 was expressed specifically on the microvilli of hepatocyte basal membranes, facing the space of Disse, where lipoprotein uptake occurs. Abundant syndecan-1 on wild-type murine hepatocytes exhibited saturable binding of VLDL and inhibition by heparin and facilitated degradation of VLDL. Furthermore, adenovirus-encoded syndecan-1 restored binding, uptake, and degradation of VLDL in isolated Sdc1-/- hepatocytes and the lipoprotein clearance defect in Sdc1-/- mice. These findings provide the first in vivo genetic evidence that syndecan-1 is the primary hepatocyte HSPG receptor mediating the clearance of both hepatic and intestinally derived triglyceride-rich lipoproteins. PMID- 19805916 TI - Ayurvedic lead poisoning: an under-recognized, international problem. PMID- 19805915 TI - Animal models of sepsis and sepsis-induced kidney injury. AB - Sepsis is characterized by a severe inflammatory response to infection, and its complications, including acute kidney injury, can be fatal. Animal models that correctly mimic human disease are extremely valuable because they hasten the development of clinically useful therapeutics. Too often, however, animal models do not properly mimic human disease. In this Review, we outline a bedside-to bench-to-bedside approach that has resulted in improved animal models for the study of sepsis - a complex disease for which preventive and therapeutic strategies are unfortunately lacking. We also highlight a few of the promising avenues for therapeutic advances and biomarkers for sepsis and sepsis-induced acute kidney injury. Finally, we review how the study of drug targets and biomarkers are affected by and in turn have influenced these evolving animal models. PMID- 19805917 TI - Recollections of learning-disabled adolescents of their schooling experiences: a qualitative study. AB - BACKGROUND: Little is known about the actual impact of the schooling experience on adolescents with specific learning disability (SpLD). AIMS: To analyze the recollections of adolescents with SpLD who were undergoing education in regular mainstream schools. SETTINGS AND DESIGN: Prospective interview-based study conducted in our clinic. MATERIALS AND METHODS: Adolescents' responses to a semi structured interview were audio-taped, transcribed verbatim and interpreted by content analysis. The number of participants was determined by 'saturation sampling,' resulting in a total of 30 adolescents. RESULTS: Twelve (40%) adolescents had 'overall' neutral recollections, 9 (30%) had 'overall' positive recollections and 9 (30%) had 'overall' negative recollections about having SpLD during their schooling. Fourteen (46.7%) adolescents stated 'getting provisions' as good features, whereas 7 (23.3%) stated 'feeling different from classmates' and 6 (20%) stated 'being teased by classmates about their disability' as bad features of having SpLD. Nineteen (63.3%) adolescents remembered classroom teachers being supportive, while 7 (23.3%) remembered being insulted by them. Of the 21 adolescents who had undergone remedial education, only 10 (47.6%) acknowledged that it had benefited them. Twenty-one (70%) adolescents stated that availing provisions helped in getting better marks in examinations. Ten (33.3%) adolescents had negative recollections about their parents' behavior in relation to their disability. Twenty (66.7%) adolescents wanted changes in their school to help students with SpLD. CONCLUSION: Improving the knowledge of classroom teachers, classmates and family members about SpLD and about the rationale of provisions will help reduce the unpleasant experiences students with SpLD undergo during their schooling years. PMID- 19805918 TI - Evaluation of lung infiltration score to predict postural hypoxemia in ventilated acute respiratory distress syndrome patients and the lateralization of skin pressure sore. AB - CONTEXT: Mechanical ventilation with positive end expiratory pressure (PEEP) is associated with unequal aeration of lungs in acute respiratory distress syndrome (ARDS) patients. Therefore, patients may develop asymmetric atelectasis and postural hypoxemia during lateral positioning. AIMS: To validate proposed lung infiltration score (LIS) based on chest x-ray to predict postural hypoxemia and lateralization of skin sores in ARDS patients. SETTINGS AND DESIGN: University hospital ICU. Prospective, observational study of consecutive patients. MATERIALS AND METHODS: Sixteen adult patients of both genders on mechanical ventilation with PEEP for 24 to <48 hours. On chest x-ray, 6 segments were identified on each lung. The proposed LIS points (0- normal; 1- patchy infiltrates; 2- white infiltrates matching heart shadow) were assigned to each segment. Without changing ventilation parameters, supine, left and right lateral positions at 45 degrees tilt were randomly changed. At the end of 20 minutes of ventilation in each position, we observed arterial oxygen saturation, hemodynamic and arterial blood gases. Later, position change protocol (4 hourly) was practiced in ICU, and skin pressure sore grading was noted within a week of ICU stay. STATISTICAL ANALYSIS USED: Nonparametric Bland and Altman correlation analysis, ANOVA and Student t test. RESULTS: Arterial oxygenation (PaO2/FiO2 = 313 +/- 145.6) was significantly (P<0.01) higher in better lung (lower LIS)-down position than supine (PaO2/FiO2 = 199 +/- 70.2) or a better lung-up position (PaO2/FiO2 = 165 +/- 64.8). The positioning-related arterial oxygenation was significant (P<0.05) at LIS asymmetry > or =3 between two lungs. CONCLUSIONS: The LIS mapping on chest x-ray was useful to differentiate between asymmetric lung disease and postural hypoxemia in ICU patients, which predisposed patients to early skin sore changes on higher LIS side. PMID- 19805920 TI - Lead-induced peripheral neuropathy following Ayurvedic medication. AB - Lead poisoning following intake of Ayurvedic medication is one of the recent areas of concern. We report a case of a 58-year-old type II diabetic man who was stable with diet control and 30 mg pioglitazone per day. He took Ayurvedic medication for generalized weakness and developed peripheral neuropathy following its intake. He was found to have high blood and urinary lead levels and was diagnosed to have subacute lead poisoning. He was treated with d-Penicillamine for 8 weeks, following which his lead levels became normal. The use of d Penicillamine was proved highly effective in treating a case of lead poisoning. PMID- 19805919 TI - Prevalence of Helicobacter pylori and cytotoxin-associated gene A in Iranian patients with non-erosive and erosive reflux disease. AB - BACKGROUND: The prevalence of Helicobacter pylori (H. pylori) infection and its virulent strain (cytotoxin-associated gene A: CagA) has not been evaluated in patients with gastroesophageal reflux disease (GERD) subgroups (erosive and non erosive) in Iran. AIMS: This study was conducted to evaluate H. pylori infection and anti-CagA status in GERD. SETTING AND DESIGN: One hundred fifty-four patients (71 male and 83 female; mean age, 42.6+/- 15.5 years) were evaluated for symptoms -heartburn and acid regurgitation--compatible with GERD. MATERIALS AND METHODS: The esophagus was diagnosed endoscopically whether it was with erosion (erosive reflux disease, ERD) or without erosion (non-erosive reflux disease, NERD). Rates of anti-H. pylori and anti-CagA-IgG (immunoglobulin G) antibodies were determined for each subject by ELISA. H. pylori was considered by ELISA if the rapid urease test proved positive. STATISTICAL ANALYSIS: anti-H. pylori and anti-CagA statuses were compared in ERD and NERD patients with and without related IgG antibodies. Mean values and P values were calculated using SPSS (version 12). RESULTS: The overall prevalence of H. pylori infection was 75.3% (116/154). Anti-CagA antibody was found in 60.3% (70/116) of infected patients. The prevalence of H. pylori infection in patients with NERD and ERD was 81.2% (56/69) and 70.6% (60/85), respectively (OR=0.56; 95% CI [confidence interval]=0.26-1.20; P=.14). The prevalence of anti-CagA antibody in infected NERD and ERD patients was 58.9% (33/56) and 61.7% (37/60), respectively (OR=1.16; 95% CI=0.49-1.70; P=0.83). CONCLUSION: The results showed that more than 70% of the patients, either NERD or ERD, acquire H. pylori infection. Anti-CagA antibody is also common in patients with reflux disorders in our community. PMID- 19805921 TI - Prenatal management with digoxin and sotalol combination for fetal supraventricular tachycardia: case report and review of literature. AB - Sustained fetal supraventricular tachycardia (SVT) with a heart rate of approximately 210 bpm may lead to increased atrial and venous pressures and congestive heart failure. There is no clear consensus regarding the best drug treatment regimens for fetal SVT. However, considerable nonrandomized experience in the transmaternal treatment of fetal SVT is available with a number of antiarrhythmic agents. We report a case of fetal supraventricular tachyarrhythmia with hydrops detected at 32 weeks that was managed with combination of oral digoxin and sotalol and review management guidelines available in the literature. PMID- 19805922 TI - Response to reference ranges for lymphocyte subsets in adults from western India: influence of sex, age and method of enumeration. PMID- 19805923 TI - Group-A beta-hemolytic streptococci resistance pattern in north Indian pediatric population. PMID- 19805924 TI - Management of nonvalvular atrial fibrillation: a comprehensive approach. AB - Atrial fibrillation is the most common arrhythmia in clinical practice, may coexist with conditions common to both cardiovascular and noncardiovascular diseases and is associated with considerable morbidity and mortality. Atrial fibrillation is often asymptomatic and diagnosed only when it has caused a potentially serious complication, such as an ischemic stroke. When atrial fibrillation has been identified, 2 objectives have to be addressed--the antiarrhythmic therapy based on rate control or rhythm control, and prevention of thromboembolism. A rhythm or rate control strategy can be chosen indifferently because they have comparable efficacy for the outcome measure of mortality, but the antithrombotic therapy is ever mandatory. The risk of stroke increases cumulatively with increasing age, previous transient ischemic attack or stroke, hypertension, diabetes mellitus, impaired left ventricular function and heart failure. Warfarin reduces the risk of stroke by about two thirds; and aspirin, by about one fifth, but its use must be weighted with the risk of bleeding. The risk of anticoagulant-associated hemorrhage increases with age, the presence of serious concomitant diseases, with poorly controlled hypertension and poorly controlled anticoagulation. PMID- 19805925 TI - Effects of shift work and intermittent noise exposure on hearing: mechanisms and prophylactic potential. AB - It is well established that intermittent noise exposure characteristically produces less hearing loss than equal energy/intensity continuous noise in animal models. Ongoing different shift work regimes open for direct studies on hearing effects of intermittent noise exposure in man without ethical concern. Amazingly, few such studies are reported. In one recent study in the present volume, noise exposed employees working 12 hours a day for two consecutive days followed by two days off, the cycle then repeated, had significantly lower permanent hearing loss than employees working nine-hour shifts from 8 am to 5 pm Monday to Friday. This commentary refers to the few studies reported, gives a short overview of the mechanisms behind noise-induced hearing loss and the protective effect of intermittent exposure, and concludes that direct studies in man on the effects of different shift work regimes on occupational hearing loss under specified noise conditions represent a prophylactic potential that calls for increased research activity. Such studies might pave the way for direct use of more optimal intermittent noise exposure regimes in future design of the noise exposure workday/-week and make future hearing conservation programs more effective. PMID- 19805926 TI - Effects of shift work on noise-induced hearing loss. AB - Evidence has accumulated concerning the adverse effects of noise on hearing acuity, but it is not clear whether working shifts may decelerate the effects of hearing loss. The objective of this study is to assess the effects of shift work on hearing loss in a noisy work environment. A sample of 218 male workers recruited at a semiconductor factory with no known occupational hazards that affected hearing acuity other than noise was chosen. The subjects worked either in an eight-hour or 12-hour shift. A standardized audiometric procedure was performed by a qualified audiologist to measure pure-tone hearing thresholds at 0.5 kHz, 1 kHz, 2 kHz, 3 kHz, 4 kHz, 6 kHz and 8 kHz in both ears. Using multiple linear regression adjusted for age, smoking habits, and work duration, the results showed that the severity of hearing loss in both ears was significantly lower in subjects who worked a 12-hour shift. In conclusion, working a 12-hour shift followed by a day off is best for workers and hearing protection should be provided in high noise areas. PMID- 19805927 TI - Development of a noise prediction model under interrupted traffic flow conditions: a case study for Jaipur city. AB - The objective of this study is to develop an empirical noise prediction model for the evaluation of equivalent noise levels (Leq) under interrupted traffic flow conditions. A new factor tendency to blow horn (AH) was introduced in the conventional federal highway administrative noise prediction (FHWA) model and a comparative study was made between FHWA and modified FHWA models to evaluate the best suitability of the model. Monitoring and modeling of Leq were carried out at four selected intersections of Jaipur city. After comparison of the results, it was found that the modified FHWA model could be satisfactorily applied for Indian conditions as it gives acceptable results with a deviation of +/-3 dB (A). In addition, statistical analysis of the data comprising measured and estimated values shows a good agreement. Hence, the modified FHWA traffic noise prediction model can be applied to the cities having similar traffic conditions as in Jaipur city. PMID- 19805928 TI - Effects of road traffic noise and irrelevant speech on children's reading and mathematical performance. AB - Irrelevant speech in classrooms and road traffic noise adjacent to schools have a substantial impact on children's ability to learn. Comparing the effects of different noise sources on learning may help construct guidelines for noise abatement programs. Experimental studies are important to establish dose-response relationships and to expand our knowledge beyond correlation studies. This experiment examined effects of road traffic noise and irrelevant speech on children's reading speed, reading comprehension, basic mathematics, and mathematical reasoning. A total of 187 pupils (89 girls and 98 boys), 12-13 years old, were tested in their ordinary classrooms. Road traffic noise was found to impair reading speed (P<0.01) and basic mathematics (P<0.05). No effect was found on reading comprehension or on mathematical reasoning. Irrelevant speech did not disrupt performance on any task. These findings are related to previous research on noise in schools and the implications for noise abatement guidelines are discussed. PMID- 19805929 TI - Sound localization with an army helmet worn in combination with an in-ear advanced communications system. AB - Conventional hearing protection devices result in decrements mainly in the ability to distinguish front from rearward sound sources. The aim of this study was to investigate the effect of wearing an earplug with advanced communications capability, in combination with an army helmet, on horizontal plane speaker identification. Ten normal-hearing male subjects were tested in a semi reverberant sound proof booth under eight conditions defined by combinations of two levels of ear occlusion (unoccluded and occluded by the earplug) and four levels of the helmet (head bare and fitted with the helmet modified to give no, partial and full ear coverage). Percent correct speaker identification was assessed using a horizontal array of eight loudspeakers surrounding the subject at one meter. These were positioned close to the midline and interaural axes of the head, at ear level. The stimulus was a 75-dB SPL, 300-ms broadband white noise. Both degree of ear coverage and ear occlusion significantly determined outcome. Overall percent correct ranged from 93.6% (bareheaded) to 79.7% (full ear coverage) with the ears unoccluded, and from 83.4%-77.5% with ear occlusion. Both variables affected the prevalence of mirror image confusions for positions 30 degrees apart in front and back of the interaural axis. With ear occlusion, front given back errors were more likely than back given front errors, increasing with degree of ear coverage to 49% and 25.4%, respectively. These errors also increased with ear coverage with the ears unoccluded, but were similar. Both degree of ear coverage and ear occlusion significantly impacted horizontal plane speaker identification, particularly for sources close to the interaural axis. However, overall percent correct was higher than observed in a previous study with conventional and level-dependent hearing protection devices, using the same array. PMID- 19805930 TI - Expert system to predict effects of noise pollution on operators of power plant using neuro-fuzzy approach. AB - Ration power plants, to generate power, have become common worldwide. One such one is the steam power plant. In such plants, various moving parts of heavy machines generate a lot of noise. Operators are subjected to high levels of noise. High noise level exposure leads to psychological as well physiological problems; different kinds of ill effects. It results in deteriorated work efficiency, although the exact nature of work performance is still unknown. To predict work efficiency deterioration, neuro-fuzzy tools are being used in research. It has been established that a neuro-fuzzy computing system helps in identification and analysis of fuzzy models. The last decade has seen substantial growth in development of various neuro-fuzzy systems. Among them, adaptive neuro fuzzy inference system provides a systematic and directed approach for model building and gives the best possible design parameters in minimum possible time. This study aims to develop a neuro-fuzzy model to predict the effects of noise pollution on human work efficiency as a function of noise level, exposure time, and age of the operators doing complex type of task. PMID- 19805931 TI - Blink rate during tests of executive performance after nocturnal traffic noise. AB - This analysis is on the hypothesis that nocturnal traffic noise affects sleep quality whereas performance decrement is avoided by increased effort expressed by a decrease in blink rates (BRs) during a visual task. Twenty-four persons (12 women, 12 men; 19-28 years, 23.56+/-2.49 years) slept during three consecutive weeks in the laboratory while exposed to road, rail, or aircraft noise with weekly permuted changes. Each week consisted of a random sequence of a quiet night (32 dBA) and three nights with equivalent noise levels of 39, 44 and 50 dBA respectively. The polysomnogram was recorded during all nights. Every morning the participants rated their sleep quality and then completed two executive tasks (Go/Nogo-, Switch-task). Neither of the two performance tests was affected by nocturnal noise. Sleep efficiency and subjective sleep quality decreased with increasing noise levels but were not associated with the type of noise. In contrast, BRs were associated with the type of noise, not with noise levels. The results do not support the hypothesis concerning the BR. The possible reasons are discussed. However, the results do not exclude that other physiological parameters such as heart rate or brain potentials measured during the tests might have revealed alterations associated with nocturnal noise exposure. PMID- 19805932 TI - Estimates of the auditory risk from outdoor impulse noise. I: Firecrackers. AB - Firecrackers are common impulse noise exposures in the United States. In this study, impulses produced outdoors by consumer firecrackers were recorded, described, and analyzed with respect to the amount of the auditory risk they pose to the unprotected listener under various listening conditions. Risk estimates were obtained using three contemporary damage risk criteria (DRC), including a waveform parameter-based approach (peak SPL and B duration), an energy-based criterion (A-weighted sound exposure level and equivalent continuous level), and a physiological model (the AHAAH model developed by Price and Kalb). Results from these DRC were converted into numbers of maximum permissible unprotected exposures to facilitate comparison. Acoustic characteristics of firecracker impulses varied with the distance, but only subtle differences were observed across firecrackers. Typical peak levels ranged between 171 dB SPL at 0.5 m and 142 dB SPL at 8 m. Estimates of the auditory risk did not differ significantly across firecrackers, but varied with the distance. Vast differences in maximum permissible exposures were observed, and the directions of the differences varied with the level of the impulse. Typical estimates of maximum permissible exposures ranged between 0 and 2 at 0.5 m and between 31 and 227,000 at 8 m. Unprotected exposures to firecracker impulses should be limited or avoided entirely if the firecrackers are ignited in batches within 8 m of the listener. Differences across DRC are inconsequential at 0.5 m, but have substantial implications at distances of 1 m and more. PMID- 19805933 TI - Estimates of auditory risk from outdoor impulse noise. II: Civilian firearms. AB - Firearm impulses are common noise exposures in the United States. This study records, describes and analyzes impulses produced outdoors by civilian firearms with respect to the amount of auditory risk they pose to the unprotected listener under various listening conditions. Risk estimates were obtained using three contemporary damage risk criteria (DRC) including a waveform parameter-based approach (peak SPL and B-duration), an energy-based criterion (A-weighted SEL and equivalent continuous level) and a physiological model (AHAAH). Results from these DRC were converted into a number of maximum permissible unprotected exposures to facilitate interpretation. Acoustic characteristics of firearm impulses differed substantially across guns, ammunition, and microphone location. The type of gun, ammunition and the microphone location all significantly affected estimates of auditory risk from firearms. Vast differences in maximum permissible exposures were observed; the rank order of the differences varied with the source of the impulse. Unprotected exposure to firearm noise is not recommended, but people electing to fire a gun without hearing protection should be advised to minimize auditory risk through careful selection of ammunition and shooting environment. Small-caliber guns with long barrels and guns loaded with the least powerful ammunition tend to be associated with the least auditory risk. PMID- 19805934 TI - Use of magnesium sulfate to manage pre-eclampsia and eclampsia in Nigeria: overcoming the odds. PMID- 19805935 TI - The use of magnesium sulphate for the treatment of severe pre-eclampsia and eclampsia. AB - BACKGROUND: Pre-eclampsia and eclampsia are important causes of maternal and perinatal morbidity and mortality in the developing countries. There is need to provide the most effective management to pre-eclamptic and eclamptic patients. There is now evidence that magnesium sulphate is the most effective anticonvulsant. METHOD: In this article , a literature review was made on the contribution of pre-eclampsia and eclampsia to maternal mortality and how it can be curtailed by the use of magnesium sulphate. RESULTS: The drug is administered by the Pritchard or Zuspan regimen, although modifications in the two protocols have been reported. CONCLUSION: A Nigerian national protocol has been developed on its use. There is need for further training of health workers on how to use this important drug. PMID- 19805936 TI - A 5-year review of maternal mortality associated with eclampsia in a tertiary institution in northern Nigeria. AB - OBJECTIVE: To determine the incidence of maternal mortality associated with eclampsia and to determine how socio-demographic and clinical characteristics of the women influence the deaths. METHODOLOGY: Records of 52 eclampsia-related mortalities from January 2003 to December 2007 were reviewed, retrospectively. Their social demography, mode and place of delivery, time of eclampsia, and fetal outcome were extracted for analysis. RESULTS: Eclampsia accounted for 52 (46.4%) of the 112 total maternal deaths recorded within the 5-year period, with case fatality of 22.33%. Age group <20, 20-29 and above 30 all had similar case fatality rate of 22.1%, 23.8% and 26.7%, respectively. Those who were experiencing their first deliveries have the worst deaths recording 42.5% of the case fatality in that category. As expected, unbooked had a higher case fatality of 24.0% compared to 15% among book cases, while those with formal education also had more death (22.3% case fatality) as compared to 3.3% among those who had some form of formal education. Antepartum eclampsia was the cause in 50% of the death, 11 (21.2%) of the pregnancies were not delivered before their death, while 18 (34.6%) were stillbirth. CONCLUSION: Eclampsia still remains the major cause of maternal mortality in this region resulting from unsupervised pregnancies and deliveries. There is need to educate and encourage the general public for antenatal care and hospital delivery. PMID- 19805937 TI - Insecticide-treated nets usage and malaria episodes among boarding students in Zaria, northern Nigeria. AB - BACKGROUND: Despite malaria being the largest public health problem in Africa South of Sahara with over one million associated deaths each year, there has been little progress in its prevention/control during the past decades. Therefore, this study was conducted to determine the knowledge, attitude, use of insecticide treated nets (ITNs), and the prevalence of malaria episodes among boarding secondary school pupils in Zaria, Nigeria. METHODS: A multi-stage sampling technique was used to sample five (5) secondary schools within Zaria, from which six classes (JSS 1 - 3 and SS 1- 3) of respondents were then randomly selected. Structured, closed-ended self-administered questionnaires were used to collect information on demographic characteristics, knowledge, attitude, and use of ITNs, reasons for non-use and malaria episodes in last 12 months. RESULTS: A total of 150 students from the five (5) boarding secondary schools were interviewed, majority were in the age group 15-17 (53.3%) with a mean (x) SD of 16.8 0.8 years. Males were 60% and females 40% of respondents. Majority of the respondents (87.3%) knew about and had actually seen an ITN; only 43.3% were current users. Most of the current users of ITN noticed a significant reduction in malaria episodes in the last 12 months. This was statistically significant (P=0.004). Cost and availability were reasons sited by non-ITN users. CONCLUSION: There is urgent need on the part of all the three tiers of Government for public health awareness campaigns through information, education and communication (IEC) to create positive ITN culture and usage. It is also suggested that ITN usage among boarding school pupils should be incorporated into school health service. PMID- 19805938 TI - The prospect of anatomy as a career choice among clinical year medical students in Nigeria. AB - BACKGROUND: Medical doctors are potential important source of anatomy teachers. The aim of this study was to determine the choice of anatomy as a career option among medical students in a Nigerian medical school METHOD: Descriptive survey of second- and third-year medical students using a self-administered questionnaire to asses age, sex, perception of anatomy as a subject, anatomy training experience and choice of anatomy as a career among respondents. Analysis was conducted using the SPSS and statistical significance inferred at P RESULTS: Three hundred and fifty three (85.3% response rate) who completely filled questionnaire were returned and analyzed of which 195 (55.2%) were males and 158(44.8%) were females. Their ages ranged between 18 and 37 years with a mean of 22.4 9.9 years. Although most students agreed that anatomy was an important subject in medical sciences 346 (98%), they had benefited from anatomy training in their clinical classes 320 (90.7%), and the knowledge of anatomy is useful in investigating patients with certain diseases 251 (71.1%), only 22 (6.2%) would choose anatomy as a career. Male students were more likely to choose anatomy as a career (P=0.026). Textbooks were the most common 334 (94.5%), while radiological means were the least 23 (6.5%) method employed in learning anatomy. CONCLUSION: Anatomy as a subject is perceived positively by clinical medical students, but the choice as a career option is low; attempt at increasing career interest is needful. PMID- 19805939 TI - Operative vaginal deliveries in Zaria, Nigeria. AB - BACKGROUND: Operative vaginal deliveries are frequent features of obstetrics practice in tertiary levels of care even in developing countries. It is essential to review these practices in order to assess their benefits or otherwise to safe motherhood in resource limited settings. STUDY DESIGN: Labor records on operative vaginal delivery cases and matched controls who had spontaneous vaginal deliveries between January 1997 and December 2001 at the Ahmadu Bello University Teaching Hospital, Zaria, Nigeria, were analyzed with respect to mode of delivery, indication for operative vaginal delivery, anesthesia use, fetal 5-min Apgar score, birth weight, fetal, and maternal complications. RESULTS: Of 7,327 deliveries at the center in the study period, 262 (3.6%) were by operative vaginal deliveries. Forceps delivery was most frequently performed (55.7%), while vacuum delivery was found to be in increased use (38.2%). Embryotomy procedures were performed selectively (6.1%). Operative vaginal deliveries were more commonly employed on primigravida (78.6%) compared to multiparas and the most common indication was delayed second stage of labor. Forcep- and vacuum-assisted deliveries were both associated with maternal and newborn complications. There was no significant difference in the use of anesthesia between forceps and vacuum deliveries. CONCLUSION: Operative vaginal delivery rates in this center are comparable to other centers as are the possible complications. Making these procedures safer will improve safe motherhood in settings where there are performed. PMID- 19805940 TI - Treatment outcomes among pulmonary tuberculosis patients at treatment centers in Ibadan, Nigeria. AB - OBJECTIVE: To assess treatment outcomes and determinants of outcome among tuberculosis patients. DESIGN: A longitudinal study design involving a cohort of sputum smear-positive pulmonary tuberculosis patients at initiation of therapy, who were followed up to the end of treatment at eighth month. SETTING: Tuberculosis treatment centers in Ibadan, Nigeria. RESULTS: A total of 1,254 patients were followed up with a mean age of 35.0+/-3.3 years. The percentages of patients with treatment outcomes assessed in the study were as follows: cure (76.6%), failure (8.1%), default (6.6%), transferred out (4.8%), and death (1.9%). The cure rate varied significantly between treatment centers from 40 to 94.4% (P<0.05). The treatment centers located within the specialist health centers at Jericho and the University College Hospital had 50 and 75% cure rates, respectively.The mean age of cured patients was 31.2+/3.1 years which was significantly lower than the mean age of those with poor treatment outcomes (36.7+/3.5 years; P<0.05). Males had a higher risk of a poor treatment outcome (RR=1.8; 95% CI: 1.02-1.94) than females. Also, patients with a poor knowledge of tuberculosis had a higher risk of having a poor treatment outcome (RR=1.35; 95% CI: 1.25-1.62) compared to those with knowledge. CONCLUSION: Variations in health center treatment outcomes and poor knowledge of tuberculosis among patients suggest that poor program implementation quality may be a major modifiable determinant of treatment outcomes in our environment. PMID- 19805941 TI - Sacrococcygeal teratoma: clinical characteristics and long-term outcome in Nigerian children. AB - BACKGROUND/PURPOSE: The excision of sacrococcygeal teratoma (SCT) may be associated with significant long-term morbidity for the child. We reviewed our experience with SCT in a tertiary health care facility in a developing country with particular interest on the long-term sequelae. METHODS: Between January 1990 and May 2008 inclusive, 38 consecutive children with the diagnosis of SCT were identified from the operation register and the Cancer Registry of the Jos University Teaching Hospital. Their clinical presentation, investigation, operative findings, histology report, and outcome were recorded and analyzed. The long-term follow-up of some of the patients were also recorded and analyzed. RESULTS: There were 31 females and 7 males. Twenty-three patients presented during the neonatal period with a median age at presentation of 7 days (range 1 18 days) and a median weight at presentation of 2.8 kg (range 2.0-3.6kg), 10 presented between 1 month and 12 months, while 5 were older than 1 year at presentation. Most of the patients had significantly external tumors. Excision of the tumor was mainly by the sacral route, four had abdominal-sacral excision. Histology was mainly benign; four were malignant at presentation. Four children with malignant disease had chemotherapy in addition to excision of the tumor. Eight had immediate post-operative wound-related complications while three children died, two of the deaths were related to anesthesia, while one died of colostomy complications. Twenty-one (60%) were followed up for a median duration of 6 years (range 1 month-8 years). Two (9.5%) had recurrent disease after primary excision; five (23.8%) had some degree of functional impairment at the follow-up. CONCLUSION: While SCT is usually benign, recurrence, malignant transformations in patients who present late and long-term functional sequelae are problems that must be tackled by the care givers. A multi-center study may be necessary to characterize this disease in developing countries and assess the long-term functional sequelae in survivors. PMID- 19805942 TI - Intrapair birthweight discordance in twins. AB - BACKGROUND: Birthweight differences in twin pairs influence their perinatal outcome as well as growth and development. Growth-discordant twins have not been well characterized in our local environment. OBJECTIVE: To determine the frequency of birthweight discordance in live-born twin pairs and document some factors that may influence it. METHODS: In this cross-sectional study, 104 live born twin pairs were recruited and their intrapair birthweight differences were determined. Twin pairs whose birthweight difference was 15% or more, using the larger infant as the growth standard, were designated as discordant. The maternal age, parity, and birth order of the heavier twin were noted. RESULTS: The prevalence of birthweight discordance was 28.8% when a 15% cut-off point was used but dropped to 9.6% when a 25% cut-off point was applied. Grand multiparity was associated with a significantly increased risk of delivery of discordant twins. Same-sex twin pairs accounted for 60% of all cases of birthweight discordance. CONCLUSION: Birthweight discordance is common in twin gestations in Benin City, especially among grand multiparous women. PMID- 19805943 TI - Prognosis of non traumatic coma: the role of some socio-economic factors on its outcome in Ibadan, Nigeria. AB - BACKGROUND: Coma occurring in the course of an illness, irrespective of cause, traditionally implies a poor prognosis and many factors may determine its outcome. These factors must be identified and possibly stratified in their order of importance. This research seeks to identify these factors and how they influenced the outcome of non-traumatic coma in our environment. METHODS: Two hundred consecutive patients, aged 18-79 years who met the inclusion criteria, the Glasgow coma scale (GCS) score of <8, history and physical findings suggestive of medical illness, no head trauma or sedation, were recruited into the study from August 2004 to March 2005 at the university College Hospital (UCH), Ibadan, after obtaining institutional ethical clearance and consent from patients' guardians. Detailed history of illness including the bio-data and time to present to the hospital and treatments given were noted. Thereafter, the clinical course of the patients was monitored daily for a maximum of 28 days during which the support of the family and/ or the hospital social welfare was evaluated. RESULTS: During the 8-month period of the study, 76% (152) of the patients died while 24% (48) survived. The following factors were associated with high mortality rate: inability to confirm diagnosis (100%), poor family support (97.1%), delay in making a diagnosis within 24 h (85.4%), poor family understanding of disease (84.1%), need for intensive care admission and management (83.3%), poor hospital social welfare support (82.4%), presentation to UCH after 6 h of coma (76.7%), and referral from private health facilities (75.7%). Others include substance abuse (100%) and seropositivity to HIV (96%) and hepatitis B surface antigen (92%) antibodies, among others. CONCLUSION: This study has demonstrated that socio-economic factors such as gender, occupation, risky lifestyle behaviors, late presentation or referral to hospital, late diagnosis and treatment, and poor family support contributed to poor outcome of nontraumatic coma. It is hoped that improvement, modification, or correction of these factors may improve coma outcome. PMID- 19805944 TI - Vesicovaginal fistula: do the patients know the cause? AB - BACKGROUND: So much has been written on vesicovaginal fistula (VVF) but there is little on the patients' perspective of the condition. The objectives of this study were to determine the knowledge of patients who have developed VVF on the causes of the fistula and their attitude toward measures that would prevent future occurrence. METHODS: The questionnaire-based survey was conducted on VVF patients on admission from June to August 2003 at Maryam Abacha Women and Children Welfare Hospital, Sokoto, Nigeria. The case notes of the patients were reviewed after the interview to match the responses from the patients with those documented in the folders. Focus group discussions were held with the maternity staff to ascertain the content and quality of existing counseling. RESULTS: One hundred and thirty patients were studied out of which 121 (93%) had no formal education. Teenagers constituted 37%, while 57% were primiparae. Thirty-five (27%) patients were divorced or separated because of the VVF. There were seven cases of recurrence after a previous successful repair. Prolonged obstructed labor was the cause of the VVF in 110 (85%) patients and 77 (70%) correctly attributed their problem to the prolonged labor. The 33 patients who could not identify the prolonged obstructed labor as the cause either attributed their condition to God/destiny or to the operation that was done to relief the obstruction and therefore would not have hospital delivery in their subsequent pregnancies. From the focus group discussions, it was confirmed that pre and post operative counseling were inadequate. CONCLUSION: Even though majority (70%) of the patients knew the cause of their fistula from the health talks, some (32%) would still not change from risky obstetric behavior. Mandatory provision of accurate and appropriate information and education to all VVF patients and their relatives or spouses by trained counselors should be ensured. Such information and education should emphasize the etiology and management of obstetric fistula in order to prevent a recurrence. PMID- 19805945 TI - Rising trend and indications of caesarean section at the university of Maiduguri teaching hospital, Nigeria. AB - OBJECTIVE: To determine the trend and indications for the use of caesarean delivery in our environment. METHOD: A retrospective review of the caesarean sections performed at University of Maiduguri Teaching Hospital from January 2000 to December 2005 inclusive. RESULTS: During the study period, there were 10,097 deliveries and 1192 caesarean sections giving a caesarean section rate of 11.8%. The major maternal indications were cephalopelvic disproportion (15.5%), previous caesarean section (14.7%), eclampsia (7.2%), failed induction of labor (5.5%), and placenta previa (5.1%). Fetal distress (9.6%), breech presentation (4.7%), fetal macrosomia (4.3%), and pregnancy complicated by multiple fetuses (4.2%) were the major fetal indications. The caesarean section rate showed a steady increase over the years (7.20% in 2000-13.95% in 2005), but yearly analysis of the demographic characteristics, type of caesarean section, and the major indications did not reveal any consistent changes to account for the rising trend except for the increasing frequency of fetal distress as an indication of caesarean section over the years, which was also not statistically significant (c[2] =8.08; P=0.12). The overall perinatal mortality in the study population was found to be 72.7/1000 birth and despite the rising rate of caesarean section, the perinatal outcomes did not improve over the years. CONCLUSION: Trial of vaginal birth after caesarean section in appropriate cases and use of cardiotocography for continuous fetal heart rate monitoring in labor with confirmation of suspected fetal distress through fetal blood acid--base study are recommended. A prospective study may reveal some of the other reasons for the increasing caesarean section rate. PMID- 19805946 TI - Mobile caecum and ascending colon syndrome in a Nigerian adult. AB - A mobile caecum and ascending colon is a rare congenital abnormality. Its presentation as a cause of right lower abdominal pain in an adult is usually mis diagnosed as acute appendicitis. A 42-year-old civil servant presented with a 2 year history of recurrent right lower quadrant pain of the abdomen. The pain was sharp in nature and persistent in the last 2 weeks and centered mainly in the right side of the abdomen. No other associated symptoms were noted. Laboratory investigations did not reveal obvious abnormality. A diagnosis of acute on chronic lower quadrant pain of unknown etiology was made. The patient was resuscitated and had exploratory laparatomy. No abnormalities were found other than the caecum and the whole ascending colon, which were unattached to the posterior peritoneum. Appendectomy and caecopexy, using a lateral peritoneal flap were performed. The diagnosis of mobile caecal syndrome should be considered in patients with chronic right lower quadrant pain, and appendectomy and caecopexy offers a great relief. PMID- 19805947 TI - Autosomal dominant polycystic kidney disease presenting with hepatic encephalopathy. PMID- 19805948 TI - Foregut cystic developmental malformation: new taxonomy and classification- unifying embryopathological concepts. AB - Foregut cystic developmental malformations are rare developmental anomalies. The problems inherent to these malformations are their presentation across specialties that include embryology, anatomy, pathology, thoracic foregut surgery, pediatric surgery and general abdominal surgery. The direct consequence of this variation has resulted in diverse terminology, classification and a failure to identify the correlation. The article aims to summarize and unify the embryological concepts of foregut cystic malformation, to suggest a generic title to the various groups of these interrelated disorders and a uniform use of nomenclature on the basis of unifying concepts of embryopathogeneis. PMID- 19805949 TI - A histopathologic study of salivary gland tumors in Kano, northern Nigeria. AB - Significant differences in the global distribution of salivary gland tumors have been reported, but no formal study has been carried out here in Kano, the largest city in northern Nigeria. We therefore undertook this eight-year retrospective study of all histologically diagnosed salivary neoplasms at the histopathology laboratory of our referral teaching hospital in Kano. Seventy-eight salivary gland tumors were diagnosed during the eight-year study period accounting for 0.4% of all neoplasms. Benign tumors were more prevalent, comprising 56.4%, while malignancies were 43.6%. Pleomorphic adenoma and mucoepidermoid carcinoma were the commonest histological types constituting 48.7% and 23.1% respectively, while the most frequent sites were parotid, submandibular and minor salivary glands accounting for 49%, 26% and 24% respectively. As in most studies of black populations there were no adenolymphomas. Age distribution was bimodal with a benign peak in the third decade and a malignant peak in the sixth. Our findings were broadly similar to most other African reports but somewhat at variance with Western literature. Improved hospital attendance with more comprehensive reporting would yield more representative data. PMID- 19805950 TI - Breast carcinoma grading, estimation of tumor size, axillary lymph node status, staging, and nottingham prognostic index scoring on mastectomy specimens. AB - INTRODUCTION: Breast carcinoma is the most common malignant tumor and the leading cause of cancer death in women. In western countries, a sharp increase in the detection of breast carcinoma, largely due to widespread use of mammography, has recently led to a fall in breast cancer mortality. This, however, is not true for less developed countries, in which mortality continues to rise. OBJECTIVE: The aim of this study was to acquire information about the extent and spread of breast carcinoma in our patients by grading the tumors, determining the tumor size, and axillary lymph node status, staging of the tumors and Nottingham Prognostic Index (NPI) scoring on the available material. MATERIALS AND METHODS: One hundred and twenty consecutive mastectomy specimens with axillary lymph node sampling removed for breast carcinoma and received in the section of histopathology, Aga Khan University, in the year 2005, were included in the study. Standard protocols were used for the processing of the specimens, and reporting was done using a standard format incorporating all relevant tumor parameters. NPI was applied to the cases. RESULTS: Out of the 120 cases, 5 (4.17) were grade 1, 91 (75.83) were grade 2, and 24 (20%) were grade 3. Also, 9 cases (7.5%) were T1 (4 were T1b, and 5 were T1c); 53 (44.16%) were T2; 50 (41.66%) were T3; and 8 (6.66%) were T4. Axillary lymph nodes were available in 107 cases. In 13 cases, no lymph nodes were recovered. Out of 107 cases 27 (25.23%) lymph nodes were negative for metastases pN0; 29 (27.10%) cases were pN1; 26 (24.30) were pN2; and 25 cases (23.36%) were pN3. Extranodal spread was present in 56 (70%) out of the 80 cases in which lymph nodes were positive. The average microscopic size of nodal metastasis was 1.7 cms. Significant statistical association was found between the number of positive nodes and perinodal extension (P = 0.001). Tumor necrosis was present in 76 out of 120 cases (63.33%). Vascular invasion was present in 43 out of 120 cases (35.83%). NPI scores were greater than 5.4 in 60 out of 107 cases (56.1%) indicating poor prognosis. CONCLUSION: The large majority of the cases were grade 2 tumors. Most cases (all grades) were T2 or T3, and were axillary lymph node positive. Large majority of cases with nodel metastases showed extra nodal spread. The majority of patients had NPI scores greater than 5.4 indicating poor prognosis. Significant statistical association was found between the number of positive nodes and perinodal extension (P = 0.001). The findings show extensive and advanced disease trends in our patients. PMID- 19805951 TI - HER-2/neu expression in lesions of uterine cervix: is it reliable and consistent? AB - This study was conducted to evaluate the expression of HER-2/neu oncogene in the lesions of the uterine cervix and to determine its correlation with histological type of malignancy, grade and clinical stage of presentation. One hundred cervical specimens were included in this study. These comprised cases with diagnosis of benign epithelial lesions, squamous cell carcinoma, adenocarcinoma, carcinoma cervix with glandular differentiation and cervical intraepithelial neoplasia. HER-2/neu immunostaining was performed by streptovidin-biotin peroxidase method. Higher expression of HER-2/neu was noted in malignant lesions as compared to benign lesions. Intensity of staining also correlated with clinical stage of presentation, lymph node metastasis and presence of parametrial extension. The over-expression of HER-2 oncoprotein is associated with poor prognosis, metastatic potential and aggressive biological behavior. PMID- 19805952 TI - Coronary atherosclerosis in sudden cardiac death: an autopsy study. AB - BACKGROUND: The incidence of ischemic heart disease (IHD) has markedly increased in India over the past few years. Considering the variations in racial, dietary and lifestyle patterns in our population, it is essential to study the biology of coronary atherosclerosis in our patients. Vulnerable plaques have a large number of foam cells, extracellular lipid, thin fibrous caps and clusters of inflammatory cells and are more prone to rupture. These plaques are nourished by the microvessels arising from the vasa vasorum of the blood vessels and by lumen derived microvessels through the fibrous cap. This autopsy study was designed to analyse the coronary arterial tree in cases of sudden cardiac death, classify coronary atherosclerotic plaques and to assess the factors contributing to vulnerability of the plaques including inflammation, calcification and microvascular density. MATERIALS AND METHODS: Seven cases of sudden cardiac death were included in the study. The hearts were perfusion-fixed and the coronary arteries along with their main branches were dissected and studied. The location of the plaques, type of plaques, presence of inflammation and calcification were assessed. The cap thickness and microvessel density per 1000 um 2 were assessed. The statistical significance was estimated. RESULTS AND CONCLUSIONS: Extensive high-grade coronary atherosclerotic disease was seen in all sudden cardiac death cases. Majority of the plaques were vulnerable. High-grade inflammation was seen in most of the vulnerable and ruptured plaques. All the ruptured plaques were uncalcified indicating that calcification probably stabilizes the plaques and protects against rupture. Increased microvessel density was noted in ruptured plaques compared to vulnerable plaques. However, it was not statistically significant. PMID- 19805953 TI - Megakaryocytic alterations in thrombocytopenia: a bone marrow aspiration study. AB - CONTEXT: Dysplastic changes are well documented in myelodysplastic syndromes (MDS). However, they are also observed in non-MDS hematological conditions. AIMS: To evaluate the megakaryocytic alterations in the bone marrow aspirations in cases of non-MDS related thrombocytopenia. SETTING AND DESIGN: A prospective study of 144 bone marrow aspirates was conducted in the department of pathology, Kasturba Medical College, Mangalore. The aspirates were studied to assess the number and morphology of the megakaryocytes in non-MDS related thrombocytopenia and evaluate their significance when compared to changes in MDS. MATERIALS AND METHODS: The bone marrow aspiration smears were stained with Leishman stain and examined under light microscope. STATISTICAL ANALYSIS USED: Fisher's exact test. A P value less than 0.05 was considered significant. Sensitivity and specificity was calculated for those features which were significant in the relevant hematological disorders. RESULTS: The sensitivity of immature megakaryocytes, dysplastic forms and micromegakaryocytes in cases of immune thrombocytopenic purpura was 100%, 89% and 42% respectively. The specificity of emperipolesis was 74%. In cases of infection-associated thrombocytopenia, immature megakaryocytes had a sensitivity of 100% and cytoplasmic vacuolization were 86% specific. The sensitivity of the dysplastic forms in megaloblastic anemia was 75%. However, no platelet budding was observed. The presence of micromegakaryocyte had a specificity of 83% in MDS, and was statistically significant when compared to cases of non-MDS conditions (P 0.05). CONCLUSIONS: Careful understanding of the morphological changes of megakaryocytes in bone marrow aspirates can improve the diagnostic accuracy for a wide range of hematological disorders thereby enabling proper therapeutic interventions. PMID- 19805954 TI - Evaluation of 100% rapid rescreening of cervical smears. AB - Advantage of rapid rescreening (RR) over conventional 10% random rescreening of all negative smears has been the topic of debate. RR of negative smears by cytopathologists/cytotechnologists have yielded good results. We undertook the rapid review of all the reported cervical smears to study its results as a means of strict quality control exercise on 2,500 smears. PMID- 19805955 TI - Predictive value of mid-trimester amniotic fluid high-sensitive C-reactive protein, ferritin, and lactate dehydrogenase for fetal growth restriction. AB - BACKGROUND: Fetal growth restriction (FGR) is surprisingly common with placental dysfunction occurring in about 3% of pregnancies and despite advances in obstetric care, FGR remains a major problem in developed countries. AIM: The purpose of this study is to find out the predictive value of amniotic fluid high sensitive C-reactive protein (hs-CRP), ferritin, and lactate dehydrogenase (LDH) for FGR. MATERIALS AND METHODS: This prospective strategy of this study has been conducted on pregnant women who underwent genetic amniocentesis between 15th and 20th weeks of gestation. All patients were followed up on until delivery. Patients with abnormal karyotype and iatrogenic preterm delivery for fetal and maternal indications were excluded. The samples were immediately sent to laboratory for cytogenetic and biochemical examination. Non-parametric tests and receiver-operator characteristic curve analysis were used for statistical purpose. RESULTS: A significant correlation between incremental amniotic fluid alpha fetoprotein (alphaFPr) and LDH levels and FGR at gestational weeks 15th 20th was found out. We also found an optimum cut-off value> 140 IU/L for the amniotic fluid LDH concentration with a sensitivity of 87.5% and a specificity of 82.4% for the prediction of FGR. CONCLUSION: Once the LDH value is confirmed, it could serve as a prediction factor for FGR at the time of genetic amniocentesis at gestational weeks 15-20. PMID- 19805956 TI - Detection and antifungal susceptibility testing of oral Candida dubliniensis from human immunodeficiency virus-infected patients. AB - CONTEXT: Candida dubliniensis, an opportunistic yeast that has been implicated in oropharyngeal candidiasis (OPC) in patients infected with Human Immunodeficiency Virus (HIV) may be under-reported due to its similarity with Candida albicans. Resistance to Fluconazole is often seen in C. dubliniensis isolates from clinical specimens. AIMS: To know the prevalence of C. dubliniensis in OPC in patients infected with HIV and their antifungal susceptibility pattern. SETTINGS AND DESIGN: One hundred and thirty-two HIV seropositive individuals and 50 healthy controls were included in the study. MATERIALS AND METHODS: Two oral swabs were collected from the site of the lesion from 132 HIV-infected patients. Oral rinse was obtained from 50 healthy controls. Samples were inoculated on Sabouraud's dextrose agar (SDA) medium and on HiCrome Candida Differential Agar (CHROM agar) medium. Isolates were speciated by standard tests. Dark green-colored, germ tube positive isolates, which failed to grow at 420C and negative for xylose assimilation were identified as C. dubliniensis. Antifungal susceptibility test was performed by Macro broth dilution technique (National Committee for Clinical Laboratory Standards guidelines). RESULTS AND CONCLUSIONS: From 132 patients, 22 (16.3%) C. dubliniensis were isolated; samples from healthy controls did not reveal their presence. Antifungal susceptibility test showed higher resistance among C. dubliniensis isolates to azoles compared to C. albicans. Five (22.7%) isolates of C. dubliniensis were resistant to Fluconazole followed by four (18.2%) to Ketoconazole. This study emphasizes the importance of identification and antifungal susceptibility testing of C. dubliniensis in HIV-infected patients. PMID- 19805957 TI - Changing trends in antimicrobial resistance of Salmonella enterica serovar typhi and salmonella enterica serovar paratyphi A in Chennai. AB - BACKGROUND AND OBJECTIVES: Chloramphenicol was considered the anti-microbial gold standard for typhoid treatment but, following the increasing worldwide frequency of antibiotic resistance, ciprofloxacin has been the mainstay of therapy since 1980. Recent studies have shown a shifting of susceptibility to conventional drugs like chloramphenicol, ampicillin and cotrimoxazole. The primary objective of the study was to evaluate the in vitro activity of chloramphenicol and other first-line drugs in comparison with cephalosporins and quinolones. MATERIALS AND METHODS: Fifty isolates of Salmonella obtained from blood culture were subjected to serotyping at the Central Research Institute, Kasauli. Phage typing and biotyping was performed at the National Phage Typing Centre, New Delhi. Antibiotic sensitivity testing was carried out for 10 drugs by the Kirby-Bauer disc diffusion method and minimum inhibitory concentration by broth microdilution for nalidixic acid, chloramphenicol, ciprofloxacin, ceftriaxone, cefixime and ofloxacin. Multi-drug-resistant (MDR) strains were checked for plasmid. RESULTS: In the present study, 70 and 30% of the isolates were Salmonella enterica serovar typhi and paratyphi A, respectively. They were highly sensitive to chloramphenicol (86%), ampicillin (84%) and cotrimoxazole (88%). Highest sensitivity was seen for cephalosporins, followed by quinolones. Seventeen/21 (81%) and 100% of the Salmonella enterica serovar typhi strains belonged to E1 phage type and biotype 1, respectively. Antibiogram showed 2% of the strains to be sensitive to all the drugs tested and 12% were MDR and showed the presence of plasmids. CONCLUSION: The study indicates reemergence of chloramphenicol susceptible Salmonella enterica serovar typhi and paratyphi A isolates, a significant decline in MDR strains and high resistance to nalidixic acid. E1 phage type and biotype 1 are found to be most prevalent in Chennai, India. PMID- 19805958 TI - Nosocomial cross-transmission of Pseudomonas aeruginosa between patients in a tertiary intensive care unit. AB - BACKGROUND: Nosocomial infection caused by Pseudomonas aeruginosa (P. aeruginosa) is very common, despite the application of various preventive measures in intensive care units (ICUs) leading to increased morbidity, mortality, prolonged hospital stay, and increased treatment cost. AIM: The aim of the present study is to identify the source of P. aeruginosa infection in patients admitted to tertiary ICU. MATERIALS AND METHODS: From 200 patients selected randomly, appropriate clinical specimens from different sites were collected and processed for the isolation and identification of the nosocomial pathogens. Surveillance samples from environmental sites and hands of nursing staff were also cultured. RESULTS: P. aeruginosa was found to be the most common pathogen associated with nosocomial infections accounting for 23.3% of all bacterial isolates from different infection sites in the ICU. Serotyping of the clinical isolates and surveillance sample isolates from nurses' hands showed serotype E as the most common serotype. Other serotypes of P. aeruginosa were isolated from environmental cultures such as sinks, floors, walls, tap water, etc. CONCLUSION: Study revealed a high prevalence of P. aeruginosa infections in the ICU attributed to cross transmission from patient to patient via hands of the nursing staff. Strict enforcement of infection control protocols is essential to minimize the disease burden. PMID- 19805959 TI - Mucinous tubular and spindle cell carcinoma of kidney: a rare case report and review of the literature. AB - Low grade mucinous tubular and spindle cell carcinoma of kidney was newly established as a distinct renal cell carcinoma in the World Health Organization (WHO) classification of 2004. Until now, less than 60 cases have been reported and the largest series represented approximately 15 patients with this type of tumor. Herein, we report a case of mucinous tubular and spindle cell carcinoma in a 63-year-old male presented with right flank pain which was diagnosed after nephrectomy. Pathologists should consider this diagnosis and its spectrum of histopathologic features in mind to ensure an accurate diagnosis. PMID- 19805960 TI - Pulmonary carcinosarcoma masquerading as a cryptic disseminated malignancy. AB - Pulmonary carcinosarcoma is an aggressive rare malignant tumor comprising a mixture of carcinoma and sarcoma components containing differentiated mesenchymal elements, such as malignant cartilage, bone, and skeletal muscle. We report a case which presented with unusual clinical features and proved cryptic until death. At autopsy, it was a stage IV lung malignancy and histopathology revealed a carcinosarcoma comprising an adenocarcinoma and an osteosarcoma with metastasis to the heart, lymph nodes, and both adrenals. To our knowledge, this is the first case of this subtype with metastasis to the heart. The present case had an unusual clinical presentation and its elusive nature towards diagnosis despite dissemination is noteworthy and unique in the literature. PMID- 19805962 TI - A case of melanoma in xeroderma pigmentosum. AB - Xeroderma pigmentosum (XP) was first described in 1874 by Hebra and Kaposi. [1] It is a rare autosomal recessive disorder characterized by photosensitivity, pigmentary changes, premature skin aging, and malignant tumor development due to cellular hypersensitivity to ultraviolet radiation resulting from a defect in DNA repair. The basic defect in XP is in nucleotide excision repair (NER), leading to deficient repair of damaged DNA. A 12-year-old boy presented with a large growth over the right side of the forehead. The lesion was first noticed before two years as a 2 x 2 cm 2 mass. It was slowly growing and attained the present size of 10 x 8 x 7 cm 3 . The surface showed ulceration with areas of hemorrhage and blackish pigmentation. Also, the patient had hyperpigmented macules over the skin since early childhood. The macules appeared initially over the face and later developed over the other areas of the body. The macules were more over the sun exposed areas. He also had photophobia and both eyes showed corneal opacities. Histopathological examination of the excised growth showed features consistent with melanoma. This case is being presented because of its rare association with xeroderma pigmentosum patients in India. PMID- 19805961 TI - Primary endobronchial synovial sarcoma confirmed by SYT-SSX1 fusion gene transcript by reverse transcriptase polymerase chain reaction. AB - Primary sarcomas of lung are rare compared to metastatic sarcomas. Herein, we report a rare case of primary pulmonary synovial sarcoma with polypoid endobronchial growth in a 35-year-old lady who presented with cough and dyspnea. A malignant pulmonary tumor was suspected and left pneumonectomy was performed. Grossly, a non-encapsulated polypoidal endobronchial tumor measuring 6 cm in greatest diameter, with a solid, tan-white cut surface was identified. Microscopically, tumor was characterized by a proliferation of oval to spindle shaped cells arranged in sheets and fascicles. Focal hemangiopericytomatous pattern was noted. Immunohistochemically, tumor cells were positive for vimentin, BCL-2, MIC-2 and calponin and focally positive for pancytokeratin and epithelial membrane antigen. A subsequent molecular analysis performed using reverse transcriptase-polymerase chain reaction with RNA extracted from paraffin-embedded tissue, revealed SYT/SSX1 fusion gene which confirmed the diagnosis of synovial sarcoma. The utility of immunohistochemistry and molecular techniques in diagnosis of such a rare case is stressed and the relevant literature is discussed. PMID- 19805963 TI - Calcifying fibrous pseudotumor in association with hyaline vascular type Castleman's disease. AB - Calcifying fibrous pseudotumor is a recently described rare entity. It is considered as sclerosing end stage of inflammatory myofibroblastic tumor. An association with hyaline vascular type Castleman's disease has also been described. We report a case of a 13-year-old boy who presented with pain in epigastrium. Computed tomography scan of abdomen revealed a circumscribed mass arising from the gastric wall along the greater curvature. Histology revealed a tumor composed of spindle cells present within the dense hyalinized collagenous tissue. Lymphoplasmacytic infiltrate was seen along with lymphoid follicles, dystrophic and ossifying calcification. Tumor cells were focally positive for alpha smooth muscle actin and negative for anaplastic lymphoma kinase protein. PMID- 19805964 TI - Nodular lymphoid hyperplasia in common variable immunodeficiency syndrome mimicking familial adenomatous polyposis on endoscopy. AB - Common variable immunodeficiency syndrome (CVID) includes a heterogeneous disorder characterized by reduced levels of IgG, IgA or IgM, and recurrent bacterial infections with normal T-cell immunity in 60% of patients. It affects the gastrointestinal tract as the largest immune organ with a wide spectrum of symptoms and signs. We present a case of nodular lymphoid hyperplasia (NLH) of the small intestine in a 31-year-old man admitted for evaluation of chronic diarrhea. Upper and lower gastrointestinal endoscopy revealed multiple polyps in the stomach, duodenum, ileum, and large intestine mimicking familial adenomatous polyposis (FAP). Although he had no history of recurrent infection, immunological profiles were in favor of CVID. We emphasize the importance of considering CVID in any patient with gastrointestinal manifestations even in the absence of recurrent bacterial infections. Diagnostic delay results in more morbidity and complications in untreated patients. PMID- 19805965 TI - Periocular metatypical cell carcinoma with scleral infiltration. AB - A 60-year-old male, laborer by occupation, presented with an ulcerative lesion of medial canthus in the left eye along with infiltration and adhesion of lids to globe. He had decreased vision and restricted extraocular movements. Exenteration with skin grafting was done. Histopathologically it was diagnosed as periocular metatypical cell carcinoma with scleral infiltration. Any suspicious lesion should be diagnosed and treated early so as to prevent such extensive damage. PMID- 19805966 TI - Coexistent gastric primary choriocarcinoma and adenocarcinoma. AB - Choriocarcinomas are commonly gestational and intrauterine or gonadal in origin. Nongestational and extragonadal disease, especially in the stomach is extremely rare with a poorer prognosis and early metastasis. We herein describe a case of primary gastric choriocarcinoma with liver metastasis in a 69-year-old male. The patient presented with abdominal pain and irregular bowel habits of one month duration. Both endoscopic and radiographic imaging revealed an ulcerated and necrotic circumferential tumor in the pyloric antrum of the stomach. The histopathologic findings of the resected tumor were that of an extensively necrotic and hemorrhagic neoplasm with combined choriocarcinomatous and adenocarcinomatous element. A strongly positive immunohistochemical staining for beta- subunit of human chorionic gonadotropin (beta-HCG) in the choriocarcinomatous component and a proper correlation with elevated serum beta HCG levels confirmed the diagnosis. PMID- 19805967 TI - Adenoid cystic carcinoma of the external auditory canal: report of two cases. AB - Adenoid cystic carcinoma occurring in the external auditory canal is very rare. Two cases are being reported. One was diagnosed on cytology based on which surgery was planned. In the second case, fine needle aspiration procedure had to be abandoned due to severe pain and a biopsy was required for its diagnosis. Morphologically, it resembles its more common salivary gland counterpart. Though severe pain during needling may be a limiting factor, adenoid cystic carcinoma of external auditory canal can be confidently diagnosed on cytology based on which surgical management can be planned. PMID- 19805968 TI - Adenoid cystic carcinoma of uterine cervix in a young patient. AB - Adenoid cystic carcinoma of uterine cervix is a rare tumor. Its origin is debatable. It has a high incidence in postmenopausal women but rarely can develop in patients under 40. An association with squamous cell carcinoma has been described. We report a case of adenoid cystic carcinoma of the endocervical canal with foci of squamous cell carcinoma in a 34-year-old suffering from menorrhagia associated with blood-stained vaginal discharge. Per vaginum and per speculum examination revealed a growth. Cervical biopsy showed bits of tissue, suggesting adenoid cystic carcinoma. Patient was operated upon and uterus with cervix sent for histopathological examination. We report this case because of its rarity, particularly in young patients, with description of illustrative pathology and discussion on the histological diagnosis. PMID- 19805969 TI - Persistent Mullerian duct syndrome. AB - Persistent Mullerian duct syndrome (PMDS) is a rare form of male pseudohermaphroditism characterized by the presence of the Momicronllerian duct structures in an otherwise phenotypically as well as genotypically normal male. We report a case of 40-year-old cryptorchid male who was clinically diagnosed as seminoma in the undescended abdominal testis. A diagnosis of PMDS was made on histological evaluation subsequent to abdominal orchidectomy. PMID- 19805970 TI - Leiomyosarcoma of the renal pelvis. AB - Leiomyosarcomas are rare malignant tumors of the kidney. They may arise from the renal capsule, renal vein, renal pelvic musculature or renal parenchyma. Renal pelvis is an uncommon site of occurrence, with around 10 cases reported in the literature so far. Here we present a 60-year-old male who presented with increased urinary frequency, lower limb weakness, anorexia and weight loss. Imaging showed a right renal mass. A renal cell carcinoma was suspected clinically. A right nephrectomy was performed, which showed a large circumscribed mass in the hilar region. Histology revealed a tumor mass arising from the renal pelvis. The tumor was composed of spindle cells arranged in fascicles. Immunohistochemistry showed tumor cells to be positive for smooth muscle actin (SMA) and desmin (Des) and negative for cytokeratin (CK), HMB 45, CD117 (C-kit), and CD34. That confirmed the diagnosis of leiomyosarcoma. PMID- 19805971 TI - Idiopathic renal replacement lipomatosis: a case report and review of literature. AB - Idiopathic renal replacement lipomatosis is a rare condition in which the renal parenchyma, especially the renal sinus is replaced with adipose tissue with increase in perirenal fat. We report a case of renal replacement lipomatosis presenting as a renal mass clinically. The computed tomography scan revealed low density and nodular masses of adipose tissue within the renal parenchyma. The differential diagnosis before surgery was angiomyolipoma, lipoma, and xanthogranulomatous pyelonephritis. After the histological examination and extensive literature search, the diagnosis of renal replacement lipomatosis was made. It is a rare entity and may be missed if not considered due to lack of experience, so we hope that more urologists, radiologists, and pathologists become aware of this entity and recognize it. PMID- 19805972 TI - Metastatic rhabdomyosarcomatous elements, mimicking a primary sarcoma, in the omentum, from a poorly differentiated ovarian Sertoli-Leydig cell tumor in a young girl: an unusual presentation with a literature review. AB - Sertoli-Leydig cell tumors (SLCTs) of the ovary with mesenchymal heterologous elements are uncommon. Only few such cases have been documented, showing presence of only mesenchymal heterologous elements at the metastatic site. We report an unusual case in a young girl who presented with an omental mass that was consistent with histopathological features of a high-grade sarcoma, with prominent rhabdomyoblastic differentiation of the embryonal type. The sections from her ovarian mass for which she was operated a year back displayed features of a poorly differentiated SLCT with heterologous elements, including focal rhabdomyoblastic differentiation. This is one of the rare cases, to the best of our knowledge, where only rhabomyosarcomatous elements were identified at the metastatic site, mimicking a primary abdominal rhabdomyosarcoma, in a case of an ovarian SLCT. Further, this case reinforces the presence of rhabdomyosarcomatous elements in an ovarian SLCT to be associated with an aggressive disease course. PMID- 19805973 TI - Granulosa cell tumor arising in an ovary with mature teratoma. AB - Granulosa cell tumor (GCT) with synchronous mature teratoma is extremely rare and only eight cases are documented in the literature. GCT is low-grade malignancy and need a close follow up for recurrences which may be late. We report a case of GCT and mature teratoma occurring synchronously in the same ovary in a perimenopausal woman. PMID- 19805974 TI - Transfusion related acute lung injury. AB - Transfusion related acute lung injury (TRALI) is an uncommon but potentially fatal adverse reaction to transfusion of plasma containing blood components. We describe a case of 10-year-old male child with aplastic anemia, platelet count of 7800/l, B positive blood group who developed fever (39.2C), difficulty in breathing and cyanosis within 2 hrs after transfusion of a random platelet concentrate. Despite the best resuscitative efforts, the child died within next 24 hrs. The present case highlights the fact that TRALI should be kept as a differential diagnosis in all patients developing acute respiratory discomfort within 6 hrs of transfusion. Without a 'gold standard' the diagnosis of TRALI relies on a high index of suspicion and on excluding other types of transfusion reactions. Notification to transfusion services is crucial to ensure that a proper investigation is carried out and at-risk donor and recipients can be identified, and risk reduction measures can be adopted. PMID- 19805975 TI - Inflammatory psuedo-tumor of the spleen. AB - Inflammatory pseudo-tumor of spleen, a benign reactive lesion of unknown etiology and pathogenesis, is extremely rare with isolated case reports in literature. These are usually misdiagnosed preoperatively, both clinically and radiologically; metastasis or lymphoproliferative disorders with pathological studies allow reliable diagnosis of the disease. We report the unusual occurrence of this lesion in the spleen. PMID- 19805976 TI - Cardiac sarcoidosis causing sudden death. AB - Sarcoidosis is a systemic disease of young adults. Cardiac involvement is rarely diagnosed clinically. In most cases it presents with arrhythmias and conduction disorders. We report a case of sudden death of a young female, wherein sarcoidosis with prominent cardiac involvement was diagnosed at autopsy. The other organs involved were lung and liver. Cardiac sarcoidosis should be considered in young patients with unexplained conduction disorders. PMID- 19805977 TI - Monophasic synovial sarcoma of tongue. AB - Synovial sarcoma is a well defined morphologic entity extensively researched in literature. Synovial sarcoma displays a wide spectrum of clinical presentations and histologic appearances that may give rise to diagnostic dilemmas. One such unusual site in the head and neck area is the tongue. We report a case of monophasic synovial sarcoma of the tongue in a 22-year-old male. Microscopically, this tumor mimicked a poorly differentiated carcinoma which is more common at this site though the patient was young for this type of tumor. On immunohistochemistry, neoplastic cells were positive for cytokeratin, vimentin, calponin, CD99 and bcl2. Molecular studies--viz. reverse transcriptase polymerase chain reaction revealed a SYT-SSX translocation clinching the diagnosis. This paper highlights the immunohistochemistry profile and SYT-SSX translocation which helped arrive at an accurate diagnosis only because the index of suspicion for a monophasic synovial sarcoma is high. PMID- 19805978 TI - Intraventricular hydatid cyst in a child. AB - Hydatid disease is caused by the infestation of the larvae of tapeworms of the genus Echinococcus. The definitive hosts of Echinococcus are various carnivores, the common being the dog. All mammals (more often sheep and cattle) are intermediate hosts. Humans get infected through the feco-oral route by ingestion of food or milk contaminated by dog feces containing ova of the parasite or by direct contact with dogs. The most common sites of infestation are the liver (75%) and lungs (15%). Various authors state a frequency of hydatidosis of the brain ranging between only 0.2-4% of cases. [1],[2],[3] The prognosis following surgical intervention is good, especially in pediatric age. [2],[3] Hence early diagnosis of this condition is crucial. We report a rare case of a child with a large intraventricular hydatid cyst that had an excellent recovery following surgery. PMID- 19805979 TI - Pancreatic abscess secondary to gall stones caused by Escherichia coli. PMID- 19805980 TI - Primary malignant melanoma of the uterine cervix. PMID- 19805981 TI - Synchronous papillary thyroid carcinoma with mucinous rectal carcinoma. PMID- 19805982 TI - Chondroid heterotopia in the fallopian tube. PMID- 19805983 TI - Sirenomelia. PMID- 19805984 TI - Objective career planning for a resident pathologist. PMID- 19805985 TI - Intra-abdominal mass: a diagnostic dilemma. PMID- 19805986 TI - Chemotherapy induced cytomorphologic changes in breast carcinoma: a potential diagnostic challenge for the histopathologist. PMID- 19805987 TI - T-cell/histiocyte rich B-cell lymphoma of mass in caecum and part of ascending colon. PMID- 19805988 TI - Alternative method of digital photography in pathology. PMID- 19805989 TI - A case of metastatic melanoma in the breast with unknown primary site, diagnosed by fine needle aspiration cytology. PMID- 19805990 TI - Longstanding lupus vulgaris with basal cell carcinoma. PMID- 19805991 TI - Partial hydatidiform mole with a full-term infant. PMID- 19805992 TI - A case of ovarian leiomyoma with cystic degeneration. PMID- 19805993 TI - A six month-old girl with botulism due to honey ingestion. PMID- 19805994 TI - Sclerosing stromal tumor of the ovary associated with benign endometrioid peritoneal implants. PMID- 19805995 TI - Mandible like structure with fourteen teeth in a benign cystic teratoma. PMID- 19805996 TI - Primary fallopian tube adenocarcinoma with brain and lung metastasis. PMID- 19805997 TI - Ciliated hepatic foregut cyst. PMID- 19805998 TI - Megakaryocytic emperipolesis: a histological finding in myelodysplastic syndrome. PMID- 19805999 TI - Associations of income and wealth with health status in the Korean elderly. AB - OBJECTIVES: This study aimed to verify the association between wealth or income level and health status after adjusting for other socio-economic position (SEP) indicators among Korean adults aged 45 and over. METHODS: Data were obtained from the 1st wave of Korean Longitudinal Study of Ageing (households: 6,171, persons: 10,254). We used self-rated health status and activities of daily living (ADLs) as dependent variables. Explanatory variables included both net wealth measured by savings, immovables, the other valuated assets and total income including pay, transfer, property and so on. Binary logistic regression was conducted to examine the relationships. Also, in order to determine the relative health inequality across economic groups, we estimated the relative index of inequality (RII). RESULTS: The inequality of health status was evident among various wealth and income groups. The wealthiest group (5th quintile) was much healthier than the poorest group, and this differential increased with age. Likewise, higher income was associated with better health status among the elderly. However, these effects, as measured by the odds ratio and RII, showed that wealth was more important in determining health status of elderly people. CONCLUSIONS: This study suggests that economic capability plays a significant role in determining the health status and other health-related problems among the elderly. Particularly, our results show that health status of the aged is related more closely to the individual's wealth than income. PMID- 19806000 TI - Assessment of community capacity building ability of health promotion workers in public health centers. AB - OBJECTIVES: This study was performed to assess the community capacity building ability of health promotion workers of public health centers and to identify influential factors to the ability. METHODS: The subjects were 43 public officers from 16 public health centers in Busan Metropolitan City. Questionnaire was developed based on 'Community Capacity Building Tool' of Public Health Agency of Canada which consists of 9 feathers. Each feather of capacity was assessed in 4 point rating scale. Univariate analysis by characteristics of subjects and multivariate analysis by multiple regression was done. RESULTS: The mean score of the 9 features were 2.35. Among the 9 feathers, 'Obtaining resources' scored 3.0 point which was the highest but 'Community structure' scored 2.1 which was the lowest. The mean score of the feathers was relatively lower than that of Canadian data. The significant influential factors affecting community capacity building ability were 'Service length', 'Heath promotion skill level', 'Existence of an executive department' and 'Cooperative partnership for health promotion'. According to the result of multiple linear regression, the 'Existence of an executive department' had significant influence. CONCLUSIONS: Community capacity building ability of subjects showed relatively lower scores in general. Building and activating an executive department and cooperative partnerships for health promotion may be helpful to achieve community capacity building ability. PMID- 19806001 TI - Workplace smoking ban policy and smoking behavior. AB - OBJECTIVES: To evaluate the impact of the workplace smoking ban in South Korea, where the male smoking rate is high (57%), on smoking behavior and secondhand smoke exposure. METHODS: A workplace smoking ban legislation implemented in April 2003 requires offices, meeting rooms, and lobbies located in larger than 3,000 square meter buildings (or 2,000 square meter multipurpose buildings) should be smoke free. A representative cross-sectional survey, the third wave (2005) of health supplements in the National Health Nutrition Survey of South Korea, was used to measure the impact of the 2003 workplace smoking ban implementation on smoking behavior. It contained 3,122 observations of adults 20 to 65 years old (excluding self-employed and non-working populations). A multivariate statistical model was used. The self-reported workplace smoking ban policy (full workplace ban, partial workplace ban, and no workplace ban) was used as the key measure. RESULTS: A full workplace smoking ban reduced the current smoking rate by 6.4 percentage points among all workers and also decreased the average daily consumption among smokers by 3.7 cigarettes relative to no smoking ban. Secondhand smoke showed a dramatic decrease of 86 percent (= -1.74/2.03)from the sample mean for full workplace ban. However, public anti-smoking campaign did not show any significant impact on smoking behavior. CONCLUSIONS: The full workplace ban policy is effective in South Korea. Male group showed bigger impact of smoking ban policy than female group. The public antismoking campaign did not show any effectiveness. PMID- 19806002 TI - [Association of blood pressure levels with carotid intima-media thickness and plaques]. AB - OBJECTIVES: The aim of this study was to investigate the association of blood pressure levels with the common carotid artery intima-media thickness (CCA-IMT) and carotid plaques. METHODS: Data were obtained from 2,635 subjects, aged 50 years and over, who participated in the Community Health Survey (a population based, cross-sectional study) in Dong-gu, Gwangju city between 2007 and 2008. Participants were categorized into three groups according to blood pressure levels; normotensives (<120/80 mmHg), prehypertensives (120-139/80-89 mmHg), and hypertensives (> or =140/90 mmHg). Prehypertensives were further categorized as low prehypertensives (120-129/80-84 mmHg) and high prehypertensives (130-139/85 89 mmHg). Carotid intima-media thickness and plaques were evaluated with a high resolution B-mode ultrasound. Statistical analyses were performed using chi square test, ANOVA, and multiple logistic regression. RESULTS: Prehypertensives had significantly greater maximal CCA-IMT values than normotensives, with a multivariate adjusted odds ratio of 1.78 (95% CI=1.36-2.32) for abnormal CCA-IMT (maximal CCA-IMT > or =1.0 mm), and 1.45 (95% CI=1.19-1.77) for carotid plaques. The multivariate adjusted odds ratio of low prehypertensives was 1.64 (95% CI=1.21-2.21) for abnormal CCA-IMT, and 1.30 (95% CI=1.04-1.63) for carotid plaques compared with normotensives. Subject with hypertension had higher frequency of abnormal CCA-IMT (odds ratio, 2.18; 95% CI=1.49-3.18), and carotid plaques (odds ratio, 1.98; 95% CI=1.46-2.67) compared with normotensives after adjustment for other cardiovascular risk factors. CONCLUSIONS: Our results indicate that there is a significant increase in the prevalence of carotid atherosclerosis in subjects with prehypertension (even in low prehypertensives) compared with normotensive subjects. Further studies are required to confirm the benefits and role of carotid ultrasonography in persons with prehypertension. PMID- 19806003 TI - [Association between the physical activity of Korean adolescents and socioeconomic status]. AB - OBJECTIVES: The physical activity of Korean adolescents and its distribution based on social characteristics have not yet been fully assessed. This study intends to reveal the distribution of physical activity by its subgroups and offer possible explanatory variables. METHODS: The 3rd Korea Youth Risk Behavior Web-based Survey was analyzed for this study. The appropriateness of physical activity was defined by Korea's Health Plan 2010 and physical inactivity was assessed independently. Family affluence scale, parents' education levels, subjective economic status, grade, and school location were considered explanatory variables. All statistical analysis was conducted using SAS ver. 9.1. RESULTS: The proportion of participants engaging in vigorous physical activity was high in males (41.6%), at a low grade (38.5%), within the high family affluence scale group (35.5%). The distribution of participants engaging in moderate physical activity showed similar patterns, but the overall proportion was lower (9.8%). Low family affluence and students with lower subjective economic status reported a higher prevalence of physical inactivity. In multiple logistic regression analysis for physical activity, significant factors included family affluence scale (p<0.05). For physical inactivity, family affluence scale, parents education levels, and subjective economic status were included as significant factors (p<0.05). CONCLUSIONS: The results suggest that the physical activity and inactivity of adolescents may be affected by socioeconomic variables, such as family affluence scale. This implies the need to take proper measures to address these socio-economic inequalities. PMID- 19806004 TI - [Management of diabetic mellitus in low-income rural patients]. AB - OBJECTIVES: Knowledge about the management status of diabetic mellitus (DM) is essential to improve diabetic management. Moreover, low income is associated with poor adherence to treatment and increased mortality. This study was performed to evaluate the management status of DM in low-income patients in a rural area. METHODS: We enrolled 370 patients with type 2 DM living in Gokseong county, JeollaNamdo. A well-trained examiner measured the height, weight, waist circumference, blood pressure, total cholesterol, triglyceride, high density lipoprotein cholesterol, fasting blood sugar and glycosylated hemoglobin (HbA1c) levels. Carotid ultrasonography was used to measure carotid artery carotid artery intima media thickness (IMT) and plaque. ankle-brachial index (ABI) was used to evaluate peripheral artery disease. A fundoscopic examination was performed to evaluate diabetic retinopathy. A history of diabetes complications and health related questionnaires were also completed. RESULTS: The age of diabetic subjects was 68.7+/-8.7 years and the duration of diabetes was 8.9+/-8.2 years. Most (63.5%) had hypertension, and 45.7% had triglycerides below 150 mg/dl, 38.1% had low density lipoprotein cholesterol (LDL) cholesterol below 100 mg/dl, 48.7% had urine albumin to creatinine ratio (UACR) below 30 mg/g. Less than half (45.9%) achieved the goal of HbA1c less than 7% suggested by the American Diabetes Association (ADA). 10.6% had peripheral vascular disease, 11.9% had retinopathy, and 60.8% had chronic kidney disease. CONCLUSIONS: DM management in low income patients is very poor and requires further work to improve. PMID- 19806005 TI - [Employment and married women's health in Korea; beneficial or harmful?]. AB - OBJECTIVES: The aim of this study was to investigate whether working married women in different occupational classes affected diverse health outcomes. METHODS: We used data for married women aged 25-59 (N=2,273) from the 2005 National Health and Nutritional Examination Survey. Outcome measures included physical/mental and subjective/objective indicators (self-rated poor health, chronic diseases, depression, and suicidal ideation from reported results; metabolic syndrome and dyslipidemia from health examination results). Age standardized prevalence and logistic regression were employed to assess health status according to three types of working groups (housewives, married women in manual jobs, married women in non-manual jobs). Sociodemographic factors (age, numbers of children under 7, education, household income) and health behaviors (health examination, sleep, rest, exercise, smoking, drinking) and a psychological factor (stress) were considered as covariates. RESULTS: Non-manual married female workers in Korea showed better health status in all five health outcomes than housewives. The positive health effect for the non-manual group persisted in absolute (age-adjusted prevalence) and relative (odds ratio) measures, but multivariate analyses showed an insignificant association of the non-manual group with dyslipidemia. Manual female workers showed significantly higher age-adjusted prevalence of almost all health outcomes than housewives except chronic disease, but the associations disappeared after further adjustment for covariates regarding sleep, rest, and stress. CONCLUSIONS: Our results suggest that examining the health impact of work on married women requires the consideration of occupational class. PMID- 19806006 TI - [Lifestyle and metabolic syndrome among male workers in an electronics research and development company]. AB - OBJECTIVES: The objectives of this study were to determine the relationship between lifestyle-implementation and metabolic syndrome in an electronics research and development company, and to provide a foundation for health providers of health management programs for setting priorities. METHODS: From July 1 to July 16, 2008 we carried out a descriptive cross-sectional survey. Consecutive workers of one R & D company in Seoul, Korea (N=2,079) were enrolled in study. A checklist for lifestyle (from the National Health Insurance Corporation) consisted of questions regarding diet, drinking, smoking and exercise. After the survey, researchers obtained data from health profiles for metabolic syndrome(waist-circumference, triglycerides, HDL cholesterol, blood pressure and fasting blood sugar level). Lifestyle was recorded as good or not good. Statistical analysis of metabolic syndrome and the lifestyle of subjects was done using multiple logistic regression analysis. RESULTS: The prevalence of metabolic syndrome in our study gropu was 13.3% (N=277). After adjustment for age, the adjusted odds ratios (odds ratio, 95% confidence intervals) for metabolic syndrome increased in proportion to the number of bad habits: two (1.72, 1.23-2.44), three (2.47, 1.73-3.56), and four (3.63, 2.03-6.34). Relative to subjects eating both vegetables and meat', the OR for 'meat' eaters was 1.66 (1.18-2.31). Compared with 'non-smokers and ever-smoker', the OR for 'current smoker' was 1.62 (1.25-2.10). Compared with 'Healthy drinker', the OR for 'unhealthy drinker' was 1.38 (1.05-1.83). CONCLUSIONS: Poor lifestyle was associated with an increased likelihood of metabolic syndrome. These findings suggest that lifestyle-based occupational health interventions for young employees should include a specific diet, smoking cessation, and healthy-drinking programs. PMID- 19806007 TI - [Prevalence of hyperhomocysteinemia and related factors in a community-based health examination survey: a cross-sectional study]. AB - BACKGROUND: Many previous studies have shown that elevated homocysteine in the serum is a well known risk factor for cardiovascular disease and this is associated with other risk factors for cardiovascular disease, but any Korean data on this is limited. OBJECTIVES: This study aimed to calculate the prevalence of hyperhomocysteinemia and to analyze the relation between elevated homocysteine and the lifestyle factors of Korean adults. METHODS: We conducted a cross sectional survey that included 650 men and 743 women (age range, 20 to 79 years) who were residents of Gwangju City in Gyeonggi-do. These subjects participated in the health interview and examination survey from November to December 2005. The total homocysteine, total cholesterol, HDL-cholesterol, LDL-cholesterol, and triglyceride in the serum were measured. All the participants had their body composition measured such as height and weight, and we obtained health-related behavioral information through the self-entry questionnaire. RESULTS: Very right handed skewed distributions of homocysteine were shown in men and women. The prevalence of hyperhomocysteinemia was 22.6% in men and 13.7% in women in Gwangju city. On the multiple logistic regression analysis, hyperhomocysteinemia was associated with age (OR=1.02, 95% CI=1.01-1.04), male gender (OR=1.60, 95% CI=1.02-2.52), severe general physical activity (OR=0.32, 95% CI=0.15-0.69) and nutrient consumption (OR=0.49, 95% CI=0.31-0.76). CONCLUSIONS: There is a great prevalence of hyperhomocysteinemia in adults of Gwangju City, Korea and it was associated with both genetic factors and lifestyle risk factors. This study can suggest that comprehensive lifestyle modification is needed in order to diminish the prevalence of hyperhomocysteinemia and to prevent CVD. PMID- 19806008 TI - Kelch-like 1 protein upregulates T-type currents by an actin-F dependent increase in alpha(1H) channels via the recycling endosome. AB - The neuronal protein Kelch-like 1 (KLHL1) is a novel actin-binding protein that modulates neuronal structure and function. KLHL1 knockout mice exhibit dendritic atrophy in cerebellar Purkinje neurons and motor dysfunction. Interestingly, KLHL1 upregulates high and low voltage-gated calcium currents (Ca(V)2.1 and Ca(V)3.2) and interacts with their respective principal subunits, alpha(1A) and alpha(1H). We reported the mechanism of enhanced Ca(V)3.2 (alpha(1H)) current density (and calcium influx) by KLHL1 is due to an increase in channel number (N) that requires the binding of actin. In this report we further elucidate the role of the actin cytoskeleton in this process using pharmacological tools to disrupt or stabilize actin filaments and to prevent protein trafficking and vesicle recycling. Disruption of the cytoskeleton did not affect the basal activity of alpha(1H), but did eliminate its modulation by KLHL1. In contrast, actin-F stabilization on its own increased basal alpha(1H) activity similar to KLHL1 but without synergy in its presence, suggesting KLHL1 requires actin-polymerization to increase alpha(1H) currents. Noise analysis revealed that actin polymerization induced an increase in N and P(o), in contrast to increased N in the presence of KLHL1. Interestingly, pharmacological or genetic disruption of endosomal recycling eliminated the increase in channel number by KLHL1 demonstrating this effect occurs via enhanced alpha(1H) re-insertion through the recycling endosome. Our findings afford insight on a novel mechanism of T-type channel modulation that could have overall functional implications for T-type channel function in the brain. PMID- 19806009 TI - From the circumsporozoite protein to the RTS, S/AS candidate vaccine. AB - The RTS,S/AS01(E) malaria vaccine candidate has recently entered Phase 3 testing. Reaching this important milestone is the culmination of more than 20 years of research and development by GlaxoSmithKline and partners and collaborators. The vaccine has been developed to protect young children and infants living in Sub Saharan Africa against clinical and severe disease caused by Plasmodium falciparum infection. Over the past 9 years, RTS,S/AS has been evaluated in multiple Phase 2 studies. The vaccine was shown to have a favorable safety profile and to be well tolerated in all age groups in which it was tested, including the intended target population of infants and young children in Sub Saharan Africa. Data obtained so far suggest that RTS,S/AS can be co-administered with other vaccines included in the routine Expanded Program of Immunization (EPI). In Phase 2 testing, the vaccine candidate was shown to confer significant protection against P. falciparum infection and clinical disease, including severe malaria. Furthermore, a trend towards an indirect beneficial effect of the vaccine on non-malarial morbidities has been observed in several trials. In this paper, we will describe the genesis of the RTS,S/AS concept, including the rationale for selecting the circumsporozoite protein (CSP) as the target antigen. Early development history of the vaccine will be briefly described. We will present the most salient results from recent Phase 2 studies conducted in the target pediatric population, which have led to the decision to progress RTS,S/AS to Phase 3 testing. If the Phase 3 results confirm the observations made during Phase 2 testing, the RTS,S/AS vaccine, when broadly implemented and judiciously integrated with other malaria-prevention measures, would have a major public health impact in Sub-Saharan Africa. PMID- 19806010 TI - Telomerase activation by c-Myc in human mammary epithelial cells requires additional genomic changes. AB - A central question in breast cancer biology is how cancer cells acquire telomerase activity required for unlimited proliferation. According to one model, proliferation of telomerase(-) pre-malignant cells leads to telomere dysfunction and increased genomic instability. Such instability leads in rare cases to reactivation of telomerase and immortalization. The mechanism of telomerase reactivation remains unknown. We have studied immortalization of cultured human mammary epithelial cells by c-Myc, a positive transcriptional regulator of the hTERT gene encoding the catalytic subunit of telomerase. Retrovirally introduced c-Myc cDNA resulted in immortalization of human mammary epithelial cells in which the cyclin dependent kinase inhibitor, p16(INK4A), was inactivated by an shRNA encoding retrovirus. However, while c-Myc introduction immediately resulted in increased activity of transiently transfected hTERT promoter reporter constructs, endogenous hTERT mRNA levels did not change until about 60 population doublings after c-Myc introduction. Increased endogenous hTERT transcripts and stabilization of telomeric DNA in cells expressing exogenous c-Myc coincided with telomere dysfunction-associated senescence in control cultures. Genome copy number analyses of immortalized cells indicated amplifications of some or all of chromosome 5, where hTERT genes are located. hTERT gene copy number, however, was not increased in one case. The results are consistent with the hypothesis that changes in chromosome 5, while not necessarily increasing hTERT gene copy number, resulted in removal of repressive chromatin structures around hTERT loci, allowing induction of hTERT transcription. These in vitro results model one possible sequence of events leading to immortalization of breast epithelial cells during cancer progression. PMID- 19806012 TI - Gain or loss of TGFbeta signaling in mammary carcinoma cells can promote metastasis. AB - The transforming growth factor beta (TGFbeta) is a potent regulator of tumor initiation, progression and metastasis. It has been known for many years that TGFbeta signaling in the carcinoma cell can suppress or promote tumor progression depending on the context of stimulation. While the impact of TGFbeta on the carcinoma cell is significant, it is now generally accepted that primary and metastatic carcinoma progression is regulated by an intricate network of host tumor cell interactions. Interestingly, recent results have revealed that gain or loss of TGFbeta signaling in carcinoma cells can promote metastasis through carcinoma cell derived TGFbeta dependent host-tumor cell interactions in vivo. Further, gain or complete abrogation of TGFbeta signaling was shown to result in gene expression signatures that correlated with poor patient prognosis in breast cancer. Specifically, the TGFbeta responsive gene expression signature correlated with poor prognosis for estrogen receptor negative (ER(-)) breast cancer while complete abrogation of TGFbeta signaling resulted in a correlation with poor outcome in lymph node positive (LN(+)) and ER(+) breast cancers. Importantly, in both cases the correlation with poor prognosis was linked to carcinoma cell derived interactions with the adjacent microenvironment. Together the current results suggest that, in addition to intrinsic carcinoma cell signaling, TGFbeta dependent host-tumor cell interactions should be considered when designing therapeutic strategies to manage carcinoma progression. PMID- 19806011 TI - Stabilization of Ca current in Purkinje neurons during high-frequency firing by a balance of Ca-dependent facilitation and inactivation. AB - Purkinje neurons fire spontaneous action potentials at ~50 spikes/sec and generate more than 100 spikes/sec during cerebellum-mediated behaviors. Many voltage-gated channels, including Ca channels, can inactivate and/or facilitate with repeated stimulation, raising the question of how these channels respond to regular, rapid trains of depolarizations. To test whether Ca currents are modulated during firing, we recorded voltage-clamped Ca currents, predominantly carried by P-type Ca channels, from acutely dissociated mouse Purkinje neurons at 30-33 degrees C (1 mM Ca). With 0.5 mM intracellular EGTA, 1-second trains of either spontaneous action potential waveforms or brief depolarizing steps at 50 Hz evoked Ca tail currents that were stable, remaining within 5% of the first tail current throughout the train. Higher frequency trains (100 and 200 Hz) elicited a maximal inactivation of <10%. To test whether this stability of Ca currents resulted from a lack of modulation or from an equilibrium between facilitation and inactivation, we manipulated the permeant ion (Ca vs. Ba) and Ca buffering (0.5 vs. 10 mM EGTA). With low buffering, Ba accelerated the initial inactivation evoked by 1-second trains, but reduced its extent at 200 Hz, consistent with an early calcium-dependent facilitation (CDF) and late calcium dependent inactivation (CDI) at high frequencies. Increasing the Ca buffer favored CDF. These data suggest that stable Ca current amplitudes result from a balance of CDF, CDI, and voltage-dependent inactivation. This modest net Ca dependent modulation may contribute to the ability of Purkinje neurons to sustain long periods of regular firing and synaptic transmission. PMID- 19806013 TI - HIF-1alpha versus HIF-2alpha in endothelial cells and vascular functions: is there a master in angiogenesis regulation? PMID- 19806014 TI - A life outside kinetochores for Bub1 kinases? PMID- 19806015 TI - Nrf2 and p21 regulate the fine balance between life and death by controlling ROS levels. PMID- 19806016 TI - Retinoid signaling regulates breast cancer stem cell differentiation. AB - The cancer stem cell (CSC) hypothesis implicates the development of new therapeutic approaches to target the CSC population. Characterization of the pathways that regulate CSCs activity will facilitate the development of targeted therapies. We recently reported that the enzymatic activity of ALDH1, as measured by the ALDELFUOR assay, can be utilized to isolate normal and malignant breast stem cells in both primary tumors and cell lines. In this study, utilizing a tumorsphere assay, we have demonstrated the role of retinoid signaling in the regulation of breast CSCs self-renewal and differentiation. Utilizing the gene set enrichment analysis (GSEA) algorithm we identified gene sets and pathways associated with retinoid signaling. These pathways regulate breast CSCs biology and their inhibition may provide novel therapeutic approaches to target breast CSCs. PMID- 19806017 TI - MYC, microRNAs and glutamine addiction in cancers. AB - The MYC oncogene encodes a transcription factor, c-Myc (Myc), which is a master regulator of cell metabolism and proliferation. Myc directly influences the expression of thousands of genes, of which, subsets are coordinately regulated with other transcription factors under specified conditions. Myc regulates entry into S phase by stimulating glucose and glutamine metabolism and mitochondrial biogenesis that are coupled to the regulation of E2F1 expression. As a transcription factor, Myc regulates genes involved in these pathways either transcriptionally or post-transcriptionally through direct regulation of microRNA expression. Myc's de-regulation of the expression of glutamine transporters and miR-23a/b that targets glutaminase, triggers an addiction to glutamine, which is required for bioenergetics, nucleotide biosynthesis and redox homeostasis in cancer cells. The induction of the miR-17 cluster by Myc attenuates E2F1 protein expression, such that interruption of this regulatory loop results in DNA replication stress. Hence, deregulated Myc expression in cancers is accompanied by key nodal points that could be exploited for therapeutic purposes. PMID- 19806018 TI - KSHV and the toll of innate immune activation. PMID- 19806019 TI - MAP4K3 enhances the expression of the BH3-only protein BID. PMID- 19806020 TI - Malaria vaccine development: an endemic country perspective. AB - The quest for an effective vaccine as an additional strategy in the control of malaria and to significantly impact the disease burden has progressed tremendously over the past decade and there is a very high probability that a malaria vaccine will be available for use in the near future. The development and deployment of an effective malaria vaccine faces several challenges and the early engagement of endemic country institutions in the entire process will be beneficial to all. It is very likely that the initial vaccine candidates will have to be improved upon, especially as our understanding of the disease processes improves and the number of potential vaccine antigens increase. This will place greater demands on the currently insufficient capacity for trials. Current strategies for testing and deployment of a malaria vaccine, for example those focusing on risk groups such as infants and children and pregnant women, will also need modifications as efforts are moved towards malaria elimination. Also, a successful malaria vaccine development process will need strengthening of the currently weak regulatory framework in endemic countries to enable them have oversight over clinical trials. The introduction of any malaria vaccine will be confronted by some cultural issues and it is essential to understand how these factors will ultimately affect its utilization. These and other challenges related to the development and deployment of an effective malaria vaccine especially as they concern endemic countries are discussed. PMID- 19806021 TI - Monitoring mitophagy in yeast: the Om45-GFP processing assay. AB - Macroautophagy (hereafter autophagy) is a ubiquitous degradative process in eukaryotic cells. Mitochondria autophagy (mitophagy) is a type of specific autophagy that degrades mitochondria selectively. Mitophagy is thought to play an important role for maintaining the quality of these organelles by eliminating damaged mitochondria, and it is involved in cellular differentiation, whereas dysfunctional mitophagy is related with neurodegenerative diseases; however, the mechanism of mitophagy is poorly understood. To facilitate the analysis of mitophagy, we recently established a simple method to monitor mitophagy in yeast, the Om45-GFP processing assay. Om45-GFP is a mitochondrial outer membrane protein. Following the uptake of mitochondria into the vacuole, Om45-GFP is degraded, releasing the intact form of GFP, which is detected by immunoblotting. Therefore, the amount of free GFP reflects the level of mitophagy. PMID- 19806022 TI - Chromosomal instability: a composite phenotype that influences sensitivity to chemotherapy. AB - Chromosomal instability (CIN) is defined as continual gain or loss of whole chromosomes or fractions of chromosomes and is a major cause of the genomic instability that characterizes most solid tumors. CIN is associated with intrinsic resistance to taxanes, acquired multidrug resistance and poor prognosis in many solid tumors, although recent evidence has shown that platinum agents, such as carboplatin, may specifically target CIN cancers. PMID- 19806023 TI - Characterization of a new cancer-associated mutant of p53 with a missense mutation (K351N) in the tetramerization domain. AB - Inactivation of the tumor suppressor p53 is central to carcinogenesis and acquisition of resistance to drug-induced apoptosis. The majority of alterations are missense mutations and occur within the DNA-binding domain. However, little is known about the point mutations in the tetramerization domain (TD). Here we investigated the properties of a new p53 mutant (Lys 351 to Asn) in the TD identified in a cisplatin-resistant ovarian carcinoma cell line (A2780 CIS). We found that K351N substitution significantly reduces the thermodynamic stability of p53 tetramers without affecting the overall half-life of the protein. Moreover, p53 K351N has a reduced ability to bind DNA and to trans-activate its specific target gene promoters, such as bax. Data obtained from the analysis of p53 subcellular localization revealed that K351N mutation inhibits the nuclear export of p53 and accumulation in the cytoplasm induced by cisplatin treatment. These results identify p53 K351N as a new cancer associated mutant with reduced tumor suppressor activity and altered functions in response to apoptotic stimuli. PMID- 19806024 TI - DNA damage foci in mitosis are devoid of 53BP1. AB - Nuclear DNA damage foci indicate ongoing DNA damage response, which is the major inducer of cell cycle arrest, cellular senescence and apoptosis. 53BP1 is one central mediator of the DNA damage response and a component of active DNA damage foci. Using an AcGFP-53BP1c fluorescent fusion protein that quantitatively reports DNA damage, we show that the recruitment of 53BP1 into gammaH2A.X containing DNA damage foci was inhibited at G(2)/M. This suggests a possible mechanism for cells to continue through the G(2) checkpoint with gammaH2A.X flagged double strand breaks via inhibition of 53BP1-mediated DNA damage signalling. PMID- 19806025 TI - p53 negatively regulates expression of FoxM1. AB - The Forkhead box M1 (FoxM1) oncogenic transcription factor is overexpressed in a majority of human tumors. p53 is a transcription factor and a major tumor suppressor that is mutated in 50% of human cancers. In this study, we compared the levels of FoxM1 in normal BJ human fibroblasts, BJ fibroblasts with p53 knockdown and corresponding BJ immortal/oncogenic cell lines with inactivated p53. We found that partial deletion or inactivation of p53 in these cells leads to upregulation of FoxM1 expression. Similarly, p53 knockdown in several human cancer cell lines with wt-p53 led to upregulation of FoxM1 mRNA and protein expression, while induction of p53 by DNA-damage led to downregulation of FoxM1. These data suggest that p53 negatively regulates FoxM1 expression and therefore inactivation of p53 in tumors could partially explain the phenomenon of FoxM1 overexpression in human cancers. PMID- 19806026 TI - Quantitative and qualitative analysis of cellular prion protein (PrP(C)) expression in bovine somatic tissues. AB - The host encoded cellular prion protein (PrP(C)) is an N-linked glycoprotein tethered to the cell membrane by a glycophosphatidylinositol (GPI) anchor. Under certain conditions, PrP(C) can undergo conversion into a conformationally-altered isoform (PrP(Sc)) widely believed to be the pathogenic agent of transmissible spongiform encephalopathies (TSEs). Understanding the tissue-specific expression of PrP(C) is crucial considering that cells expressing high levels of PrP(C) bear a risk for conversion and accumulation of PrP(Sc). In the present study, fifteen bovine somatic tissues were analyzed for PrP(C) expression by quantitative western blot and immunohistochemistry. Quantitative western blot analysis revealed highest expression of PrP(C) in cerebellum, obex and spinal cord. Intermediate levels were detected in thymus, intestine, nerve, heart and spleen, and lower levels in lung, muscle, kidney, lymph node, skin, pancreas and liver. Immunohistochemical analysis detected intense cellular-specific PrP(C) staining in neurons, thymocytes and lymphocytes. PrP(C) was also detected in the enteric wall, pancreatic islets of langerhans, myocardium, pulmonary alveolar sacs, renal glomeruli and dermal epithelial cells. This study demonstrated the quantitatively varied, wide-spread, tissue- and cell-specific expression pattern of PrP(C) in bovine somatic tissues. The importance of this study is to lay the foundation for understanding the tissue-specific expression of PrP(C) and to consider the potential participation of more bovine tissues in the transmission of BSE infection. PMID- 19806027 TI - KV4.3 expression and gating: S2 and S3 acidic and S4 innermost basic residues. AB - Effects of neutralizing individual negatively charged (acidic [E,D]) and innermost positively charged (basic [K,R]) residues in transmembrane segments S2 (D230Q, E240Q), S3 (D263Q) and S4 (K299A/Q, R302A/Q) of the K(V)4.3 putative voltage sensing domain (VSD) were determined. S2 D230Q generated large macroscopic currents, depolarized steady-state activation ("a(4)") and isochronal (1 sec) inactivation ("i") relationships, and significantly accelerated kinetics of deactivation and recovery (from both macroscopic and closed state inactivation [CSI]). D230Q thus stabilized non-inactivated closed states. These effects were attributable to structural perturbations, and indicated D230 is not primarily involved in voltage sensing. Both S2 E240Q and S3 D263Q failed to generate measurable ionic currents, suggesting deletion of negative charges at these putatively more intracellular acidic positions were functionally "lethal" to macroscopic K(V)4.3 function. S4 innermost positive charge deletion mutants K299A/Q and R032A/Q generated functional currents with reduced peak amplitudes. While reduced K299A/Q and R302A/Q currents prevented accurate determination of "a(4)" and estimates of potential electrostatic perturbations, both sets of mutants: (i) depolarized potentials at which currents could be macroscopically detected, consistent with stabilization of closed states (structural perturbations); and (ii) accelerated macroscopic recovery. These results provide further evidence that: (i) basic residues in S4 are involved not only in regulating K(V)4.3 activation and deactivation, but also CSI and recovery; and (ii) suggest putative electrostatic interactions between acidic S2/S3 and basic S4 residues may be different in K(V)4.3 from those proposed to exist in Shaker. Functional implications are discussed. PMID- 19806028 TI - DNA damage induced Pol eta recruitment takes place independently of the cell cycle phase. AB - When DNA is damaged in cells progressing through S phase, replication blockage can be avoided by TLS (Translesion DNA synthesis). This is an auxiliary replication mechanism that relies on the function of specialized polymerases that accomplish DNA damage bypass. Intriguingly, recent evidence has linked TLS polymerases to processes that can also take place outside S phase such as nucleotide excision repair (NER). Here we show that Pol eta is recruited to UV induced DNA lesions in cells outside S phase including cells permanently arrested in G(1). This observation was confirmed by different strategies including global UV irradiation, local UV irradiation and local multi-photon laser irradiation of single nuclei in living cells. The potential connection between Pol eta recruitment to DNA lesions outside S phase and NER was further evaluated. Interestingly, the recruitment of Pol eta to damage sites outside S phase did not depend on active NER, as UV-induced focus formation occurred normally in XPA, XPG and XPF deficient fibroblasts. Our data reveals that the re-localization of the TLS polymerase Pol eta to photo-lesions might be temporally and mechanistically uncoupled from replicative DNA synthesis and from DNA damage processing. PMID- 19806029 TI - Perturbation of survivin expression affects chromosome alignment and spindle checkpoint in mouse oocyte meiotic maturation. AB - Survivin is a member of inhibitors of apoptosis proteins (IAPs), which have multiple regulatory functions in mitosis, but its roles in meiosis remain unknown. Here, we report its expression, localization and functions in mouse oocyte meiosis. Survivin displayed a maximal expression level in GV stage, and then gradually decreased from Pro-MI to MII stages. Immunofluorescent staining showed that survivin was restricted to the germinal vesicle, associated with centromeres from pro-metaphase I to metaphase I stages, distributed at the midzone and midbody of anaphase and telophase spindles, and located to centromeres at metaphase II stages. Depletion of survivin by antibody injection and morpholino injection resulted in severe chromosome misalignment, precocious polar body extrusion, and larger-than-normal polar bodies. Overexpression of survivin resulted in severe chromosome misalignment and prometaphase I or metaphase I arrest in a large proportion of oocytes. Our data suggest that survivin is required for chromosome alignment and that it may regulate spindle checkpoint activity during mouse oocyte meiosis. PMID- 19806030 TI - Enhancement of cancer chemotherapy by simultaneously altering cell cycle progression and DNA-damage defenses through global modification of the serine/threonine phospho-proteome. AB - Despite improvements in the therapeutic efficacy of rationally designed cancer treatment regimens, most cancers remain incurable once spread beyond their sites of origin. Failure to achieve sustained control or eradication of cancers arises in large part because a subpopulation of quiescent "cancer stem cells" is insensitive to drugs targeting cell growth and replication and because defense mechanisms critical to survival of the normal cell also protect the cancer cell from cytotoxic injury. Global alteration of signal transduction by inhibition of serine/threonine dephosphorylation has recently been shown to markedly potentiate cancer cell killing by the DNA-methylating drug, temozolomide. Inhibition of the multifunctional protein phosphatase 2A appears to drive quiescent cancer cells into cycle and simultaneously inhibits cycle arrest, permitting cancer cell entry into mitosis despite the presence of chemotherapy induced DNA-damage. Absence of toxicity in animal models suggests that multi-site mutations in pathways regulating cell cycle in cancer cells make them more vulnerable than normal cells to large changes in the balance of phosphorylation-regulated signaling. Global modulation of the serine-threonine phospho-proteome may be a general method for enhancing the effectiveness of cytotoxic cancer therapy. PMID- 19806031 TI - Homology between DUF784, DUF1278 domains and the plant prolamin superfamily typifies evolutionary changes of disulfide bonding patterns. PMID- 19806032 TI - Astrin regulates meiotic spindle organization, spindle pole tethering and cell cycle progression in mouse oocytes. AB - Astrin has been described as a microtubule and kinetochore protein required for the maintenance of sister chromatid cohesion and centrosome integrity in human mitosis. However, its role in mammalian oocyte meiosis is unclear. In this study, we find that Astrin is mainly associated with the meiotic spindle microtubules and concentrated on spindle poles at metaphase I and metaphase II stages. Taxol treatment and immunoprecipitation show that Astrin may interact with the centrosomal proteins Aurora-A or Plk1 to regulate microtubule organization and spindle pole integrity. Loss-of-function of Astrin by RNAi and overexpression of the coiled-coil domain results in spindle disorganization, chromosome misalignment and meiosis progression arrest. Thr24, Ser66 or Ser447 may be the potential phosphorylation sites of Astrin by Plk1, as site-directed mutation of these sites causes oocyte meiotic arrest at metaphase I with highly disordered spindles and disorganized chromosomes, although mutant Astrin localizes to the spindle apparatus. Taken together, these data strongly suggest that Astrin is critical for meiotic spindle assembly and maturation in mouse oocytes. PMID- 19806033 TI - Visualizing the mapped ion pathway through the Na,K-ATPase pump. AB - The Na(+),K(+)-ATPase pump achieves thermodynamically uphill exchange of cytoplasmic Na(+) ions for extracellular K(+) ions by using ATP-mediated phosphorylation, followed by autodephosphorylation, to power conformational changes that allow ion access to the pump's binding sites from only one side of the membrane at a time. Formally, the pump behaves like an ion channel with two tightly coupled gates that are constrained to open and close alternately. The marine agent palytoxin disrupts this coupling, allowing both gates to sometimes be open, so temporarily transforming a pump into an ion channel. We made a cysteine scan of Na(+),K(+)-ATPase transmembrane (TM) segments TM1 to TM6, and used recordings of Na(+) current flow through palytoxin-bound pump-channels to monitor accessibility of introduced cysteine residues via their reaction with hydrophilic methanethiosulfonate (MTS) reagents. To visualize the open-channel pathway, the reactive positions were mapped onto a homology model of Na(+),K(+) ATPase based on the structure of the related sarcoplasmicand endoplasmic reticulum (SERCA) Ca(2+)-ATPase in a BeF(3)(-)-trapped state,(1,2) in which the extra-cytoplasmic gate is wide open (although the cytoplasmic access pathway is firmly shut). The results revealed a single unbroken chain of reactive positions that traverses the pump from the extracellular surface to the cytoplasm, comprises residues from TM1, TM2, TM4 and TM6, and passes through the equivalent of cation binding site II in SERCA, but not through site I. Cavity search analysis of the homology model validated its use for mapping the data by yielding a calculated extra-cytoplasmic pathway surrounded by MTS-reactive residues. As predicted by previous experimental results, that calculated extra-cytoplasmic pathway abruptly broadens above residue T806, at the outermost end of TM6 that forms the floor of the extracellular-facing vestibule. These findings provide a structural basis for further understanding cation translocation by the Na(+),K(+) ATPase and by other P-type pumps like the Ca(2+)- and H(+),K(+)-ATPases. PMID- 19806035 TI - Using 18F-fluorodeoxyglucose positron emission tomography to monitor clinical outcomes in patients treated with neoadjuvant chemo-radiotherapy for locally advanced pancreatic cancer. AB - BACKGROUND: Pancreatic cancer ranks as the fourth leading cause of cancer death in the United States with 5-year survival ranging from 1% to 5%. Positron emission tomography (PET) is a metabolic imaging system that is widely used for the initial staging of cancer and detecting residual disease after treatment. There are limited data, however, on the use of this molecular imaging technique to assess early tumor response after treatment in pancreatic cancer. METHODS: The objective of the study was to explore the relationship of early treatment response using the F-fluorodeoxyglucose (FDG) PET with surgical outcome and overall survival in patients with locally advanced pancreatic cancer. FDG-PET measurements of maximum standardized uptake value and kinetic parameters were compared with the clinical outcome. RESULTS: Twenty patients were enrolled in the study evaluating neoadjuvant induction chemotherapy followed by concurrent chemoradiotherapy (chemo-RT) for locally advanced pancreatic cancer. All 20 patients had prestudy PET scans and a total of fifty PET scans were performed. Among patients who were PET responders (> or =50% decrease in standardized uptake value after cycle 1), 100% (2/2) had complete surgical resection. Only 6% (1/16) had surgical resection in the PET nonresponders (<50% decrease). Two patients did not have the second PET scan because of clinical progression or treatment toxicity. Mean survival was 23.2 months for PET responders and 11.3 months for nonresponders (P = 0.234). Similar differences in survival were also noted when response was measured using Patlak analysis. CONCLUSIONS: FDG-PET can aid in monitoring the clinical outcome of patients with locally advanced pancreatic cancer treated with neoadjuvant chemo-RT. FDG-PET may be used to aid patients who could have complete surgical resection as well as prognosticate patients' survival. PMID- 19806034 TI - Towards revealing the structure of bacterial inclusion bodies. AB - Protein aggregation is a widely observed phenomenon in human diseases, biopharmaceutical production, and biological research. Protein aggregates are generally classified as highly ordered, such as amyloid fibrils, or amorphous, such as bacterial inclusion bodies. Amyloid fibrils are elongated filaments with diameters of 6-12 nm, they are comprised of residue-specific cross-beta structure, and display characteristic properties, such as binding with amyloid specific dyes. Amyloid fibrils are associated with dozens of human pathological conditions, including Alzheimer disease and prion diseases. Distinguished from amyloid fibrils, bacterial inclusion bodies display apparent amorphous morphology. Inclusion bodies are formed during high-level recombinant protein production, and formation of inclusion bodies is a major concern in biotechnology. Despite of the distinctive morphological difference, bacterial inclusion bodies have been found to have some amyloid-like properties, suggesting that they might contain structures similar to amyloid-like fibrils. Recent structural data further support this hypothesis, and this review summarizes the latest progress towards revealing the structural details of bacterial inclusion bodies. PMID- 19806036 TI - A phase I study of capecitabine, irinotecan, celecoxib, and radiation as neoadjuvant therapy of patients with locally advanced rectal cancer. AB - OBJECTIVES: We conducted a prospective phase I trial to determine the maximum tolerated dose of capecitabine and irinotecan when used in combination with celecoxib and radiation as preoperative therapy for patients with locally advanced rectal cancer. METHODS: Patients with histologic diagnosis of adenocarcinoma of distal rectum, evidence of T3/T4 tumor or nodal involvement by endorectal ultrasound/magnetic resonance imaging, any T status where tumor was close to but not involving the sphincter requiring abdominoperineal resection were evaluated by standard phase I methodology. Starting chemotherapy dosage (dose level: 0) was capecitabine 550 mg/m bid, day 1 to 5 every week through out x-ray therapy, irinotecan 30 mg/ m IV on days 1, 8, 22, 29 (no treatment on day 15 and day 36), and celecoxib 400 mg PO bid from day 1 till the last day of radiation. Radiation dosage of 50.4 Gy in 28 fractions was delivered in 5.6 weeks. If no dose limiting toxicity was observed, dose of capecitabine was escalated by 75 mg/m and irinotecan by 5 mg/m. Celecoxib dosage was fixed. RESULTS: Fourteen patients were accrued. Dose limiting toxicity was observed at level 2 and was primarily hematological and gastrointestinal. Two patients at level 2 developed grade-3 diarrhea and thrombocytopenia and 1 patient at level 2 developed grade 3/4 vomiting, diarrhea and dehydration. CONCLUSIONS: Recommended dosage for future trials is capecitabine 625 mg/m bid, irinotecan 35 mg/m, and celecoxib 400 mg orally bid in combination with pelvic radiation. PMID- 19806037 TI - Postoperative radiotherapy for cervix cancer incidentally discovered after a simple hysterectomy for either benign conditions or noninvasive pathology. AB - OBJECTIVE: Report the long-term outcome of patients who received postoperative radiotherapy for incidentally discovered cervix cancer following simple hysterectomy. METHODS: We recorded tumor status, treatment complications, and survival of 25 patients treated at our institution from 1961 to 2004 with postoperative RT for invasive cervix cancer discovered following simple hysterectomy (median follow-up, 17 years). All patients had stage IA2-II squamous cell carcinoma (76%) or adenocarcinoma (24%) of the cervix. RESULTS: One patient had an isolated vaginal-cuff recurrence and was cured long-term with salvage surgery. No patient died of cervix cancer. The actuarial rate of tumor control and relapse-free survival at 5, 10, and 20 years was 96%. One patient died of a treatment-related complication. Cause-specific survival was 100% at 5 and 10 years, but 92% at 20 years. Overall survival was 100% at 5 years, 95% at 10 years, and 62% at 20 years.The complications rate from therapy was surprising. The overall grade 2 to 5 complications rate was 36% (9 of 25). Twenty percent (5 of 25) of patients experienced grade 4 or 5 complications. CONCLUSIONS: This series demonstrates the price we pay for adding comprehensive radiotherapy to simple hysterectomy for early-stage cervix cancer. The findings support 2 recommendations: (1) Avoid postoperative radiotherapy by aggressively screening patients for invasive disease before performing simple hysterectomy. (2) Raise the threshold for delivering pelvic radiotherapy following simple hysterectomy with an incidental diagnosis of invasive cervix cancer. We recommend vaginal brachytherapy alone in patients with negative postoperative imaging, negative surgical margins, and <10 mm tumor invasion. PMID- 19806038 TI - The effect of intravenous contrast on photon radiation therapy dose calculations for lung cancer. AB - OBJECTIVE: The aim of this study was to evaluate the effect of intravenous contrast-enhanced computed tomography (CT) scans on the photon radiation dose calculations for lung cancer treatment planning. MATERIALS AND METHODS: Nonionic iodinated intravenous contrast (Iohexol) was administered during the treatment planning CT scan of 9 patients with node-positive non-small-cell lung cancer (NSCLC). The potential effect of intravenous contrast was studied by changing the density of the contrast-enhanced vessels. A total of 9 patients were treated in this study: 5 patients with intensity-modulated radiation therapy (IMRT), and 4 patients with three-dimensional (3D) conformal radiation therapy. A treatment plan was generated from an unmanipulated "normal contrast" planning scan. The same planning parameters were then applied to a "no contrast" planning scan. The effect of intravenous contrast was quantified by calculating the percent change of dose in a variety of target and normal structures. To evaluate a worst-case scenario, the comparison between "normal contrast" and "no contrast" planning scans was repeated, assigning each vessel the artificial high density of 1.3 g/cm. RESULTS: Dose differences between the planning image set using intravenous contrast and the image set without contrast were less than 2.5% for planning target volumes. A worst-case scenario in which normal contrast was overridden with an artificially high density of 1.3 g/cm led to small dose differences of less than 3%. CONCLUSIONS: Planning lung radiation therapy treatment using CT scans that contain intravenous contrast does not result in clinically significant errors in dose delivery. PMID- 19806039 TI - The world's experience with facial transplantation: what have we learned thus far? AB - The objective of this review article is to summarize the published details and media citations for all seven face transplants performed to date to point out deficiencies in those reports so as to provide the basis for examining where the field of face transplantation stands, and to act as a stimulus to enhance the quality of future reports and functional outcomes. Overall long-term function of facial alloflaps has been reported satisfactorily in all seven cases. Sensory recovery ranges between 3 and 6 months, and acceptable motor recovery ranges between 9 and 12 months. The risks and benefits of facial composite tissue allotransplantation, which involves mandatory lifelong immunosuppression analogous to kidney transplants, should be deliberated by each institution's multidisciplinary face transplant team. Face transplantation has been shown thus far to be a viable option in some patients suffering severe facial deficits which are not amenable to modern-day reconstructive technique. PMID- 19806040 TI - The use of a retroauricular fascioperichondrial flap in the recreation of the antihelical fold in prominent ear surgery. AB - The surgical treatments for prominent ears are on the basis of the approach over the basics of the deformity decrease the concha-mastoid angle and create a new antihelical fold. The author describes a new retroauricular flap to be used in the recreation of the antihelical fold and critically analyzes the cosmetic results and complications. A skin ellipse is outlined on the posterior surface of the ear. After hydrodissection with anesthetic solution in the outlined area, the skin is removed as thin as possible over the superior mid ear, preserving the underlying connective tissue and perichondrium. A triangular flap is created in the superior third of the raw area and elevated in the subperichondrial plane. The fixation of the flap posteriorly allows the medialization of the upper pole, resulting in a natural antihelical fold. One hundred sixty-one otoplasties were performed in 82 patients (3 unilateral), 53 women and 29 men. Mean age was 19.2 years and mean follow-up time was 3.4 years. Loss of correction at the upper pole led to secondary correction in 12 ears (7.45%). Suture extrusion was observed in 7 ears (4.34%). Hypertrophic scarring developed in 3 ears (1.86%). The accumulated risk of reoperation was of 10.85% in 9 years. The retroauricular fascioperichondrial flap combined with other techniques may be used as a standard procedure in the surgical treatment of prominent ears. PMID- 19806041 TI - Serratus anterior composite flaps for reconstruction of large-area oral and maxillofacial defects: a new neuromuscular flap. AB - Large area defects in the maxillofacial field are difficult to restore with sufficient esthetic and functional outcome. This study was to assess the feasibility of using serratus anterior composite flaps for reconstruction of large-area oral and maxillofacial tissue defects and to determine subsequent effects on neuromuscular function. Six patients with severe maxillofacial deformities were treated with serratus anterior composite flaps. The neurovascularized pedicle of each flap retained the long thoracic nerve to supply motor innervation. The long thoracic nerve was anastomosed with a branch of the facial nerve. The remaining fine branches of the facial nerve were separated interfascicularly and intrafascicularly, and then implanted within the border of the serratus anterior muscle. Transplanted flaps survived in all patients without vascular crisis. At the 12-month follow-up examinations, patients were able to close their eyes, wrinkle their forehead, and blow through pursed lips using the reconstructed mimetic muscles. The free serratus anterior composite flap has different digitations of the serratus anterior and motor nerve innervations, which is an attractive choice for reconstruction of large defects of oral and maxillofacial tissue. The nerve anastomosis and facial nerve replantation technique may protect the pedicle from atrophy of the transferred flap. Facial motion can be animated by neurotization or reinnervation in-growth in the neurotized slips of the serratus. PMID- 19806042 TI - Surgical treatment of hermaphroditism: experience with 25 cases. AB - This article is a summary of our experience with surgical treatment of 25 patients with hermaphroditism. We treated 25 patients with hermaphroditism since 1985, including 12 with male pseudohermaphroditism, 9 with female pseudohermaphroditism and 4 with true hermaphroditism. Decision on sex reassignment for these patients was made according to their genetic sex, gonad sex, social sex, psychologic sex, and the request of the patients and their relatives. Of the 12 male pseudohermaphrodites, 1 was reassigned with the male sex and penile reconstruction was performed, and the other 11 had a female sex assignment and received undescended testis removal, clitoral reduction, labioplasty of the labia minora and labium majus, and vaginoplasty. Nine female pseudohermaphrodites with female sex assignment underwent clitoral reduction, labioplasty of the labia minora and labium majus, and vaginoplasty. The 4 true hermaphrodites also had female sex assignment, and received such procedures as resection of the ovotestes and undescended testes, clitoral reduction, and labioplasty of the labia minora; resection of the uterus and appendices was also performed in 1 case due to the identification of malignant cells. The reconstructed penis in the patient with male sex assignment had good appearance and allowed normal urination. In patients with female sex assignment, the reconstructed external genitalia had also good appearance and good sensation without obvious contraction. No serious complications occurred in these patients, who were satisfactory with the outcome and reported stable gender identity, and 3 married patients reported normal sexual life. Plastic surgery is still the primary option for correction of hermaphroditism after determination of sex assignment, and satisfactory effect can be achieved by surgical intervention with stable gender identity and minimal complications. PMID- 19806043 TI - Role of subcutaneous abdominal fat on cardiac function and proinflammatory cytokines in premenopausal obese women. AB - The role of surgically removing subcutaneous fat by abdominoplasty on circulating inflammatory markers and myocardial dysfunction, evaluated by myocardial performance index (MPI), were investigated. Twenty volunteers submitted to the abdominoplasty (abdominoplasty group), and other 28 women treated by hypocaloric diet (diet group) were evaluated. Echocardiographic parameters of MPI, circulating levels of tumor necrosis factor (TNF)-alpha and interleukin (IL)-6, were performed at baseline and 2 months later. Compared with nonobese women, obese women had increased concentrations of TNF-alpha (P < 0.01), IL-6 (P < 0.01), and higher MPI (P < 0.02), indicating ventricular dysfunction. Subcutaneous fat concentrations of TNF-alpha and IL-6 were related to MPI impairment. After 60 days, waist-to hip ratio was significantly reduced in the abdominoplasty group. Anthropometric changes were accompanied by a significant decline in plasma concentrations of TNF-alpha and IL-6 levels as well as by significant improvements of MPI in abdominoplasty group compared with diet group. Abdominoplasty may represent a safe method for ameliorating cardiac function in obese women. PMID- 19806044 TI - Vertical and horizontal coordinates of the nipple-areola complex position in males. AB - After massive weight loss, male breasts in particular are one of the most disturbing body regions and can be a difficult area to treat often requiring nipple-areola repositioning. The need for bilateral repositioning of the nipple areola complex is also increasing in other surgical fields such as in female-to male transsexual patients with large breasts and in patients with severe forms of gynecomastia. Proper configuration and localization of the nipple-areola complex requires both meticulous planning and a thorough understanding of the male anatomy and is essential in determining final aesthetic outcome. Currently available guidelines create areolas that are too large, place the nipple-areola complex too high and too far medially, and/or require complex abstract mathematical calculations. Relying on the recently appreciated aesthetic value of the golden number Phi (phi) we propose an easy and reliable method to determine the horizontal and vertical coordinates of the male nipples. With only 2 easily measurable distances, umbilicus-anterior axillary fold apex and umbilicus suprasternal notch, the internipple distance and the position of the horizontal nipple plane relative to the suprasternal notch can be calculated. The internipple distance can be determined with 95% accuracy and the distance from the suprasternal notch can be determined in 80% of cases within a range of 3.33 +/- 1.25 cm. PMID- 19806045 TI - An evaluation of calcium hydroxylapatite (Radiesse) for cosmetic nasolabial fold correction: a meta-analysis and patient centric outcomes study. AB - The use of calcium hydroxylapatite (Radiesse, BioForm Medical, San Mateo, California) for cosmetic soft tissue augmentation has increased significantly in recent years. Only a handful of authors have reported patient satisfaction data. A meta-analysis of published data was performed using a 5-point scale for patient satisfaction to allow data comparison across studies. A patient-centric outcomes study was also performed at our institution, using a blinded questionnaire. Five usable studies were identified for meta-analysis. Patient satisfaction was 4.16 of 5 in 324 patients at 3 to 6 months and 4.15 in 86 patients at 1 year. In our institutional patient-centric outcomes study, the early satisfaction rate was 3.7 of 5. At 1 year the rate was only 2.3 of 5. Calcium hydroxylapatite remains a viable option for augmentation of facial soft tissue structures. However, this report shows that further, accurate, large population outcomes assessments are absolutely required. PMID- 19806046 TI - Retrospective analysis of two hundred thirty-five pediatric mandibular fracture cases. AB - Maxillofacial fractures are encountered less commonly during childhood period due to anatomic, social, cultural, and environmental factors. Although the incidence of all maxillofacial fractures is 1% to 15% among pediatric and adolescent patients, this rate drops to less than 1% in children below 5 years age. Two hundred thirty-five cases (0.7 indicates acceptable agreement). Pathologists' estimation of micro- and macrosteatosis disclosed also poor correlation (ICC: 0.22, 0.55, respectively). Inconsistent assessment of histological features of steatohepatitis (lobular inflammation, portal inflammation, hepatocyte ballooning, and Mallory hyaline) was documented. Poor conformity was also shown between the computerized quantification and ratings of 3 pathologists (Spearman rank correlation coefficients: 0.22, 0.82, 0.28, and 0.38). CONCLUSION: Quantification of hepatic steatosis in histological sections is strongly observer-dependent, not reproducible, and does not correlate with the computerized estimation. Current standards of assessment, previously published data and the clinical relevance of hepatic steatosis for liver surgery and transplantation must be challenged. PMID- 19806054 TI - Pancreatic cyst fluid protein expression profiling for discriminating between serous cystadenoma and intraductal papillary mucinous neoplasm. AB - BACKGROUND: Many patients with benign serous cystadenoma (SCA) of the pancreas will undergo resection because of the inability to reliably discriminate between SCA and premalignant mucinous cysts (intraductal papillary mucinous neoplasm [IPMN], mucinous cystic neoplasm [MCN]). METHODS: Cyst fluid from patients with SCA (n = 15), non main-duct and noninvasive IPMN (n = 32), and noninvasive MCN (n = 12) was aspirated at the time of operative resection and analyzed. Commercially available and custom designed multiplex assays (Luminex) were performed using a biomarker panel developed for pancreatic cancer. Differential protein expression (fluorescence intensity, FI) was compared between the 3 groups for each protein (Wilcoxon rank sum test). Unsupervised sample clustering (hierarchical clustering) and supervised sample classification (prediction analysis for microarrays [PAM]) was then performed. RESULTS: Differential protein expression (P < 0.05) was identified between SCA and IPMN (34/51 proteins, 67%) and between SCA and MCN (13/51 proteins, 25%). The majority of proteins were down-regulated in IPMN and MCN compared with SCA. The only proteins significantly overexpressed in the cyst fluid of patients with mucinous cysts were CEA (median FI: IPMN 11.4, MCN 13.0, SCA 5.3; P < 0.001, IPMN vs. SCA) and CA72.4 (median FI: IPMN 10.4, MCN 10.5, SCA 9.9; P = 0.003, IPMN vs. SCA). Unsupervised cluster analysis demonstrated distinct clustering of SCA and IPMN with some cross-over between MCN. Supervised sample classification with 14 proteins had an overall accuracy rate of 92% between SCA and IPMN. CONCLUSIONS: In this study differential cyst fluid protein expression was observed between SCA and IPMN for the majority of proteins assessed and multimarker sample classification accurately discriminated between SCA and IPMN in 92% of patients. PMID- 19806056 TI - One hour hypothermic oxygenated perfusion (HOPE) protects nonviable liver allografts donated after cardiac death. AB - OBJECTIVES: To test, in a large animal model, the efficacy of machine perfusion to rescue livers after prolonged ischemic injury. BACKGROUND: Our group previously showed in various rodent models the benefit of endischemic hypothermic oxygenated perfusion (HOPE) in protecting liver injury from donation after cardiac death (DCD). Convincing results are needed in large animal models before application in human. METHODS: A new model of DCD liver transplantation in large pigs was developed. Pig livers (1300 +/- 210 g each) were harvested 60 minutes after induction of cardiac death (respirator withdrawal). In situ flush and organ procurement were initiated without heparin pretreatment. Then, livers were preserved for 7 hours in cold Celsior (DCD-group) prior to orthotopic transplantation (OLT). Some livers were treated by 1 hour HOPE prior to implantation (HOPE-group). In a first step, animals were kept under anesthesia for 6 hours after orthotopic transplantation. Endpoints included serum (AST) and tissue (ATP, glutathione) markers of injury, bile flow, and histology. In a second step, survival experiments were performed. RESULTS: Livers from the DCD group displayed diffuse necrosis of hepatocytes, increased adhesion of platelets, high AST release, absence of bile flow, depletion of glutathione, and ATP. In contrast, livers treated with HOPE showed dramatic reduction of necrosis, platelet adhesion, while bile flow, ATP recovery and glutathione were improved. Importantly, untreated DCD livers caused graft failure and death of all recipients within 6 hours of reperfusion, whereas HOPE treated DCD livers remained hemodynamically stable. CONCLUSIONS: This is the first study in a reliable large animal transplant model demonstrating the efficacy of a simple cold oxygenated machine perfusion system to rescue, otherwise lethal, ischemic injured DCD liver grafts. PMID- 19806057 TI - Laparoscopic major hepatectomy: an evolution in standard of care. AB - OBJECTIVE: To analyze the results of 6 international surgical centers performing laparoscopic major liver resections. SUMMARY BACKGROUND DATA: The safety and feasibility of laparoscopy for minor liver resections has been previously demonstrated. Major anatomic liver resections, initially considered to be unsuitable for laparoscopy, are increasingly reported by several centers worldwide. METHODS: Prospective databases of 3 European, 2 U.S., and 1 Australian centers were combined. Between 1997 and 2008, 210 major liver resections were performed: 136 right and 74 left hepatectomies. Results and differences in surgical techniques between the 6 centers are outlined. RESULTS: Surgical duration was 250 minutes (range: 90-655 minutes). Operative blood loss was 300 mL (range: 20-2500 mL). Thirty patients (14.3%) received blood transfusion. Conversion to open surgery was required in 26 patients (12.4%). Portal triad clamping was performed in 24 patients (11.4%). Median tumor size was 5.4 cm (range: 1-25 cm) and surgical margin was 10.5 mm (range: 0-70 mm). Two patients died during the postoperative period from pulmonary embolism and urosepsis. Liver specific and general complications occurred in 17 (8.1%) and 29 patients (13.8%), respectively. Hospital length of stay was 6 days (range: 1-34 days). A further analysis of early (n = 90) and late (n = 120) experience showed improved surgical and postoperative results in the latter group. CONCLUSIONS: This multicenter study demonstrates that laparoscopic major liver resections are feasible in selected patients and results improve with experience. However, proficiency in both open liver surgery and advanced laparoscopy is compulsory and surgeons must begin with minor laparoscopic resections. PMID- 19806058 TI - Minimally invasive liver resection for metastatic colorectal cancer: a multi institutional, international report of safety, feasibility, and early outcomes. AB - OBJECTIVE: To evaluate a multicenter, international series on minimally invasive liver resection for colorectal carcinoma (CRC) metastasis. SUMMARY BACKGROUND DATA: Multiple single series have been reported on laparoscopic liver resection for CRC metastasis. We report the first collaborative multicenter, international series to evaluate the safety, feasibility, and oncologic integrity of laparoscopic liver resection for CRC metastasis. METHODS: We retrospectively reviewed all patients who underwent minimally invasive liver resection for CRC metastasis from February 2000 to September 2008 from multiple medical centers from the United States and Europe. The multicenter series of patients were accumulated into a single database. Patient demographics, preoperative, operative, and postoperative characteristics were analyzed. Actuarial overall survival was calculated with Kaplan-Meier analysis. RESULTS: A total of 109 patients underwent minimally invasive liver resection for CRC metastasis. The median age was 63 years (range, 32-88 years) with 51% females. The most common sites of primary colon cancer were sigmoid/rectum (51%), right colon (25%), and left colon (13%). Synchronous liver lesions were present in 11% of patients. For those with metachronous lesions liver lesions, the median time interval from primary colon cancer surgery to liver metastasectomy was 12 months. Preoperative chemotherapy was administered in 68% of cases prior to liver resection. The majority of patients underwent prior abdominal operations (95%). Minimally invasive approaches included totally laparoscopic (56%) and hand-assisted laparoscopic (41%), the latter of which was employed more frequently in the US medical centers (85%) compared with European centers (13%) (P = 0.001). There were 4 conversions to open surgery (3.7%), all due to bleeding. Extents of resection include wedge/segmentectomy (34%), left lateral sectionectomy (27%), right hepatectomy (28%), left hepatectomy (9%), extended right hepatectomy (0.9%), and caudate lobectomy (0.9%). Major liver resections (> or =3 segments) were performed in 45% of patients. Median OR time was 234 minutes (range, 60-555 minutes) and blood loss was 200 mL (range, 20-2500 mL) with 10% receiving a blood transfusion. There were no reported perioperative deaths and a 12% complication rate. Median length of hospital stay for the entire series was 4 days (range, 1 22 days) with a shorter stay in medical centers in the United States (3 days) versus that seen in Europe (6 days) (P = 0.001). Negative margins were achieved in 94.4% of patients. Actuarial overall survivals at 1-, 3-, and 5-year for the entire series were 88%, 69%, and 50%, respectively. Disease-free survivals at 1-, 3-, and 5-year were 65%, 43%, and 43%, respectively. CONCLUSIONS: Minimally invasive liver resection for colorectal metastasis is safe, feasible, and oncologically comparable to open liver resection for both minor and major liver resections, even with prior intra-abdominal operations, in selected patients and when performed by experienced surgeons. PMID- 19806059 TI - On the distinction between interaction and effect modification. AB - This paper contrasts the concepts of interaction and effect modification using a series of examples. Interaction and effect modification are formally defined within the counterfactual framework. Interaction is defined in terms of the effects of 2 interventions whereas effect modification is defined in terms of the effect of one intervention varying across strata of a second variable. Effect modification can be present with no interaction; interaction can be present with no effect modification. There are settings in which it is possible to assess effect modification but not interaction, or to assess interaction but not effect modification. The analytic procedures for obtaining estimates of effect modification parameters and interaction parameters using marginal structural models are compared and contrasted. A characterization is given of the settings in which interaction and effect modification coincide. PMID- 19806060 TI - Estimating direct effects in cohort and case-control studies. AB - Estimating the effect of an exposure on an outcome, other than through some given mediator, requires adjustment for all risk factors of the mediator that are also associated with the outcome. When these risk factors are themselves affected by the exposure, then standard regression methods do not apply. In this article, I review methods for accommodating this and discuss their limitations for estimating the controlled direct effect (ie, the exposure effect when controlling the mediator at a specified level uniformly in the population). In addition, I propose a powerful and easy-to-apply alternative that uses G-estimation in structural nested models to address these limitations both for cohort and case control studies. PMID- 19806062 TI - The effects of visual acuity on target discrimination and shooting performance. AB - PURPOSE: To explore the effects of visual acuity on target discrimination and marksmanship. METHODS: A randomized, single-blind, repeated measures study was conducted to evaluate the effect of visual acuity on target discrimination and marksmanship. Subjects were examined for inclusion criteria and equivalent plus lens over-refractions necessary to obtain the five test visual acuity ranges. Qualified subjects fired at randomized target presentations from 50 to 300 m while wearing customized spectacles to yield the visual acuity levels being evaluated. Subjects then rotated foxholes for target discrimination sequences. Randomized targets marked as friend or enemy were presented in like manner for each of the five visual acuity levels. RESULTS: Subjects at acuities better than 0.2 LogMAR were significantly more likely to hit the target than those with acuities equal to or worse than 0.7 LogMAR. Target discrimination results show subjects with acuities between -0.2 and 0.6 LogMAR were significantly more likely to correctly identify the target than those with acuities equal to or worse than 0.7 LogMAR. In addition, those at acuities of 0.0 LogMAR or better performed significantly better than those with acuities between 0.3 and 0.6 LogMAR. CONCLUSIONS: Visual acuity significantly affects target discrimination and marksmanship performance. Results of this study closely mirror expected findings based on visual angles with acuities in the range 0.3 to 0.4 LogMAR (20/40-20/50) being the transition zone where performance begins to deteriorate. This performance-based evidence fully supports a vision classification system that mandates eyewear to maximize visual performance for deployable military members. Based on the results of this study, the current U.S. Army, U.S. Navy, and U.S. Marine Corps vision readiness standards of 20/40 or better uncorrected or corrected vision seems to be a valid choice because it is the transition point for performance in both target discrimination and marksmanship ability. PMID- 19806061 TI - Weight change, initial BMI, and mortality among middle- and older-aged adults. AB - BACKGROUND: It is not known how the relationship between weight change and mortality is influenced by initial body mass index (BMI) or the magnitude of weight change. METHODS: We use the nationally representative Health and Retirement Study (n = 13,104; follow-up 1992-2006) and Cox regression analysis to estimate relative mortality risks for 2-year weight change by initial BMI among 50- to-70-year-old Americans. We defined small weight loss or gain as a change of 1-2.9 BMI units and large weight loss or gain as a change of 3-5 BMI units. RESULTS: Large and small weight losses were associated with excess mortality for all initial BMI levels below 32 kg/m (eg, hazard ratio [HR] for large weight loss from BMI of 30 = 1.61 [95% confidence interval = 1.31-1.98]; HR for small weight loss from BMI of 30 = 1.19 [1.06-1.28]). Large weight gains were associated with excess mortality only at high BMIs (eg, HR for large weight gain from BMI of 35 = 1.33 [1.00-1.77]). Small weight gains were not associated with excess mortality for any initial BMI level. The weight loss-mortality association was robust to adjustments for health status and to sensitivity analyses considering unobserved confounders. CONCLUSIONS: Weight loss is associated with excess mortality among normal, overweight, and mildly obese middle- and older-aged adults. The excess risk increases for larger losses and lower initial BMI. These results suggest that the potential benefits of a lower BMI may be offset by the negative effects associated with weight loss. Weight gain may be associated with excess mortality only among obese people with an initial BMI over 35. PMID- 19806063 TI - Collagen type distribution in the healthy human tympanic membrane. AB - BACKGROUND: A weakening of the collagen fibers of the lamina propria of the tympanic membrane is a prerequisite for the formation of a retraction pocket. The various collagen types differ in tensile strength. The distribution of the different collagen types in the lamina propria of the healthy human tympanic membrane has not been reported before. METHODS: Immunohistochemical staining for collagen Types I, II, III, and IV in healthy human tympanic membranes harvested during translabyrinthine surgery for acoustic neuroma. The staining was semiquantified using light microscopy. RESULTS: Collagen Type II was the most abundant collagen of the lamina propria. When subdividing the staining between the 2 fiber layers of the lamina propria, it was observed that the inner layer was enriched with Type III relative to Type II, as reflected in staining patterns. In the outer radial fiber layer of the lamina propria, staining for collagen Type II was predominant.The fibrous annulus could be subdivided into an inner and an outer portion by immunohistochemistry. The inner portion stained strongest for collagen Type II and to a lesser extent for Type I. The outer portion stained strongest for collagen Type III and Type I. CONCLUSION: The differences in distribution of collagen types in the different fiber layers of the lamina propria suggest that the lattice of connective tissue supporting the tympanic membrane is not uniform. Understanding the differences in collagen type distribution and in the physical properties of the individual collagen types themselves may contribute to a comprehensive model of retraction pocket pathogenesis. PMID- 19806064 TI - Laterality of exostosis in surfers due to evaporative cooling effect. AB - OBJECTIVES: 1. To correlate exostosis severity with ear canal evaporative cooling. 2. To assess hearing and complications after canalplasty. STUDY DESIGN: Retrospective chart review. SUBJECTS AND METHOD: A retrospective chart review from 1990 to 2007 at a university tertiary referral center. RESULTS: Surfers from the west coast of the United States were twice as likely to have severe exostoses in the right ear compared with the left. Evaporative cooling from a predominant northerly wind direction during the coldest water temperature months in this region may contribute to this lateral bias because surfers on this coast spend most of their time facing west. Few postoperative complications were identified. No cases of facial nerve injury or entry into the temporomandibular joint occurred. Differences in preoperative versus postoperative pure-tone hearing thresholds were not significant. CONCLUSION: Exostosis severity seems to correspond to the ear that is more exposed to the predominant coastal wind. We propose that evaporative cooling in a cold water environment contributes to greater progression of exostoses in the ear with more exposure to the predominant wind. Exostosis removal using the postauricular approach carries a low complication rate. PMID- 19806065 TI - Anterior cervical discectomy and fusion in the ovine model. AB - Anterior cervical discectomy and fusion (ACDF) is the most common surgical operation for cervical radiculopathy and/or myelopathy in patients who have failed conservative treatment. Since the operation was first described by Cloward and Smith and Robinson in 1958, a variety refinements in technique, graft material and implants have been made. In particular, there is a need for safe osteoinductive agents that could benefit selected patients. The ovine model has been shown to have anatomical, biomechanical, bone density and radiological properties that are similar to the human counterpart, the most similar level being C3/4. It is therefore an ideal model in which preclinical studies can be performed. In particular this methodology may be useful to researchers interested in evaluating different devices and biologics, including stem cells, for potential application in human spinal surgery. PMID- 19806066 TI - 'The market does not solve social problems. Public health does.'. PMID- 19806067 TI - How to find nothing. AB - Hypothesis testing can be misused and misinterpreted in various ways. Limitations in the research design, for example, can make it almost impossible to reject the null hypothesis that a policy has no effect. This article discusses two examples of such experimental designs and analyses, in which, unfortunately, the researchers touted their null results as strong evidence of no effect. PMID- 19806068 TI - Assessment of HIV/AIDS related risks among men having sex with men (MSM) in Addis Ababa, Ethiopia. AB - Ethiopia has a high prevalence of HIV/AIDS. Single-point estimates show adult HIV prevalence of 2.1 per cent for 2007. The 2005 Ethiopia Demographic and Health Survey (EDHS 2005) had indicated a national-level adult prevalence rate of 1.4 per cent among adults aged between 15 and 49 years (6 and 0.7 per cent in urban and rural residents, respectively). The rate in the capital, Addis Ababa, was 5 per cent. We conducted an ethnographic study to explore the possible role of men having sex with men (MSM) in the epidemiology of HIV/AIDS in the city of Addis Ababa, Ethiopia. To collect data, we used four focus group discussions with MSM, six key informant interviews, and extended observations at different places in Addis Ababa. These data were analyzed using open code software and a 'grounded theory' approach. We found that contextual factors, such as sociocultural background and politicolegal situation, predisposed to HIV/AIDS-related risk behavior among members of the study subgroup. The infection might have been transmitted between the study population and heterosexual people in the community by many possible routes. The study showed that men have sex with men among a segment of Addis' population. The portion of the population involved is non negligible. They are exposed to HIV/AIDS risks. It is, therefore, high time that the issue be openly discussed in the context of current efforts to control the HIV pandemic. PMID- 19806069 TI - Commentary: Ethiopia's salvation: Pragmatism in public health policy-making. PMID- 19806070 TI - Factors associated with serum retinol, alpha-tocopherol, carotenoids, and selenium in Hispanics with problems of HIV, chronic hepatitis C, and drug use. AB - The effects of hepatitis and drug use on nutritional problems in HIV infection have rarely been examined despite the importance of drug use in the global HIV pandemic. We examined the effects of HIV, hepatitis C, and drug use on serum micronutrients in 300 US Hispanic adults. Chronic hepatitis C infection was associated with lower serum retinol (-8.2 microg/dl, P < 0.0001), alpha tocopherol (-0.10 ln microg/dl, P = 0.024), and carotenoids (-19.8 microg/dl, P < 0.0001). HIV infection was associated with lower selenium (-6.1 microg/l, P = 0.028). Elevated triglycerides in HIV infection were associated with higher serum retinol and alpha-tocopherol. Drug use was not independently associated with micronutrient alterations. We conclude that hepatitis C is an important determinant of low serum micronutrients, and should be considered in any nutritional assessment of HIV infected populations. As the safety of micronutrient supplementation is not established, policy for appropriate HIV clinical care should distinguish between populations with and without hepatitis coinfection. PMID- 19806072 TI - The impact of traumatic stressors in civilian occupational settings. AB - We use the literature reporting prevalence and aetiology of post-traumatic stress disorder (PTSD) in first responders as a catalyst to discuss for organisations the policy implications for prevention and intervention of psychiatric morbidity. We searched PubMed and Google to identify studies and reports of mental health and behavioural problems in occupations including police, fire, and emergency service workers. The prevalence of PTSD ranged from 6 per cent to 32 per cent. Biological markers of PTSD, such as neuroendocrine activity, appear less useful than psychological markers, such as levels of hostility and self-efficacy, to predict PTSD. Prevalence of PTSD was generally less than that found among victims themselves, but higher than general community prevalence. Theoretically, if prevention and intervention strategies were working effectively, there should be a minimal rate of psychiatric morbidity attributable to these individuals' workplaces. Against this background, there is a case for routine screening on an annual basis for those at risk. PMID- 19806071 TI - Breast feeding: A time to craft new policies. AB - New studies of breastfeeding have discovered or confirmed the benefits to mother and child. They reinforce an emphasis on exclusive breastfeeding - no other food or fluids - during the first 6 months. Studies include findings from across the world, in well-resourced and poorly resourced settings. They also emphasize longer duration of breastfeeding, into the second year of life, and gradual rather than abrupt weaning. For HIV-infected mothers, the dangers of non exclusive feeding in the first half year of life have been well documented in recent publications. Other studies open up the possibilities for antiretroviral treatment to accompany breastfeeding, whether given to the mother, or child, or both. To be effective, implementation of any recommendations must consider individual, family, and community resources. PMID- 19806073 TI - Controlling infectious disease outbreaks: Lessons from mathematical modelling. AB - Epidemiological analysis and mathematical models are now essential tools in understanding the dynamics of infectious diseases and in designing public health strategies to contain them. They have provided fundamental concepts, such as the basic and effective reproduction number, generation times, epidemic growth rates, and the role of pre-symptomatic infectiousness, which are crucial in characterising infectious diseases. These concepts are outlined and their relevance in designing control policies for outbreaks is discussed. They are illustrated using examples from the 2003 severe acute respiratory syndrome outbreak, which was brought under control within a year, and from pandemic influenza planning, where mathematical models have been used extensively. PMID- 19806074 TI - Epidemiological characteristics of HIV-infected women in the Republic of Korea: A low HIV prevalence country. AB - Greater than 90 per cent of HIV-infected Koreans are men, and therefore the majority of Korean HIV/AIDS studies reflect the male population. We wished to evaluate the characteristics of HIV infection in women, and to compare our results with those in other countries. To better elucidate HIV infection in Korean women, we examined the characteristics of heterosexual transmission and studied the dynamics of HIV infection in heterosexual couples. We evaluated 479 HIV-infected women who were registered with the government during 1985-2007. The median age at registration and median CD4 cell count were 37 years (N = 479), 362 cells/mm(3) (N = 276), respectively. About 68 per cent had a regular sexual partner and 26 per cent were diagnosed because they had HIV-positive male partners. Eighty-five per cent of women of concordant couples were detected because of previously diagnosed male partners (N = 115). We verified that the majority of HIV-infected women were infected by male sexual partners. These results suggest that early and active detection of HIV-positive men will facilitate earlier detection and prevention in women. PMID- 19806075 TI - WFPHA: World Federation of Public Health Associations: www.wfpha.org. PMID- 19806079 TI - Inhibition of nitric oxide synthesis during induced cholestasis ameliorates hepatocellular injury by facilitating S-nitrosothiol homeostasis. AB - Cholestatic liver injury following extra- or intrahepatic bile duct obstruction causes nonparenchymal cell proliferation and matrix deposition leading to end stage liver disease and cirrhosis. In cholestatic conditions, nitric oxide (NO) is mainly produced by a hepatocyte-inducible NO synthase (iNOS) as a result of enhanced inflow of endotoxins to the liver and also by accumulation of bile salts in hepatocytes and subsequent hepatocellular injury. This study was aimed to investigate the role of NO and S-nitrosothiol (SNO) homeostasis in the development of hepatocellular injury during cholestasis induced by bile duct ligation (BDL) in rats. Male Wistar rats (200-250 g) were divided into four groups (n=10 each), including sham-operated (SO), bile duct-ligated (BDL), tauroursodeoxycholic acid (TUDCA, 50 mg/kg) and S-methylisothiourea (SMT, 25 mg/kg) treated. After 7 days, BDL rats showed elevated serum levels of gamma glutamiltranspeptidase, aspartate aminotransferase, alanine aminotransferase, LDH, and bilirubin, bile duct proliferation and fibrosis, compared with the SO group. TUDCA treatment did not significantly alter these parameters, but the iNOS inhibitor SMT ameliorated hepatocellular injury, as shown by lower levels of circulating hepatic enzymes and bilirubin, and a decreased grade of bile duct proliferation and fibrosis. Both TUDCA and SMT treatments reversed Mrp2 canalicular pump expression to control levels. However, only SMT treatment significantly lowered the increased levels of plasma NO and S-nitrosation (S nitrosylation) of liver proteins in BDL rats. Moreover, BDL resulted in a reduction of the S-nitrosoglutathione reductase (GSNOR/Adh5) enzymatic activity and a downregulation of the GSNOR/Adh5 mRNA expression that was reverted by SMT, but not TUDCA, treatment. A total of 25 liver proteins, including S-adenosyl methionine synthetase, betaine-homocysteine S-methyltransferase, Hsp90 and protein disulfide isomerase, were found to be S-nitrosated in BDL rats. In conclusion, the inhibition of NO production during induced cholestasis ameliorates hepatocellular injury. This effect is in part mediated by the improvement of cell proficiency in maintaining SNO homeostasis. PMID- 19806080 TI - Potentiation of a functional autoantibody in narcolepsy by a cholinesterase inhibitor. AB - We have recently reported the presence of an immunoglobulin G (IgG) autoantibody (Ab) in patients with narcolepsy with cataplexy that abolishes spontaneous colonic migrating motor complexes (CMMCs) and increases smooth muscle tension and atropine-sensitive phasic contractions in a physiological assay of an isolated colon. In this study, we used the cholinesterase inhibitor, neostigmine, to explore the mechanism of the narcoleptic IgG-mediated disruption of enteric motor function in four patients with narcolepsy with cataplexy and to identify a pharmacological mimic of the Ab. Neostigmine potentiated the narcoleptic IgG mediated increase in smooth muscle resting tension and phasic smooth muscle contractions by an atropine-sensitive mechanism but exerted no effect on resting tension in the presence of control IgG. Decreased frequency of CMMCs mediated by IgG with anti-M3R activity was reversed by neostigmine. Therefore, a challenge with a cholinesterase inhibitor improves the specificity of the CMMC assay for narcoleptic IgG. Tetrodotoxin (TTX), a neuronal sodium channel blocker, also abolished CMMCs and increased resting tone, and a similar potentiation was observed with neostigmine; thus, TTX is a mimic of the functional effects of the narcoleptic IgG in this bioassay. These findings provide a link to pharmacological studies of canine narcolepsy and are consistent with a functional blockade of both excitatory and inhibitory motor neurons by the narcoleptic Ab, similar to the TTX mimic, presumably by binding to an autoantigenic target expressed in both populations of neurons. PMID- 19806083 TI - The endoscopic pancreatic function test. PMID- 19806081 TI - Timp-3 deficiency impairs cognitive function in mice. AB - Extracellular matrix (ECM) degradation is performed primarily by matrix metalloproteinases (MMPs). MMPs have recently been shown to regulate synaptic activity in the hippocampus and to affect memory and learning. The tissue inhibitor of metalloproteinase (Timp) is an endogenous factor that controls MMP activity by binding to the catalytic site of MMPs. At present, four Timp isotypes have been reported (Timp-1 through Timp-4) with 35-50% amino-acid sequence homology. Timp-3 is a unique member of Timp proteins in that it is bound to the ECM. In this study, we used the passive avoidance test, active avoidance test, and water maze test to examine the cognitive function in Timp-3 knockout (KO) mice. Habituation was evaluated using the open-field test. The water maze test showed that Timp-3 KO mice exhibit deterioration in cognitive function compared with wild-type (WT) mice. The open-field test showed decreased habituation of Timp-3 KO mice. Immunostaining of brain slices revealed the expression of Timp-3 in the hippocampus. In situ zymography of the hippocampus showed increased gelatinolytic activity in Timp-3 KO mice compared with WT mice. These results present the first evidence of Timp-3 involvement in cognitive function and hippocampal MMP activity in mice. Moreover, our findings suggest a novel therapeutic target to be explored for improvement of cognitive function in humans. PMID- 19806084 TI - NOTES and other emerging trends in gastrointestinal endoscopy and surgery: the change that we need and the change that is real. AB - In this inaugural year of a historic presidency, gastroenterologists and gastrointestinal surgeons may well want to turn their attention to more immediate transformative events that have the potential to revolutionize their own practice in the near future. The most visible and, perhaps, controversial of these is natural orifice transluminal endoscopic surgery (NOTES), but other equally important changes are emerging as investigators around the globe vie with one another in the demonstration of increasingly audacious procedures. As is to be expected, we are also already seeing a backlash from more conservative scholars attempting to temper what they believe to be the surgical equivalent of irrational exuberance. However, by far the most common attitude among gastroenterologists toward these changes is one of indifference. In this piece, we discuss the circumstances that led to the development of NOTES and other innovative procedures, the peril that lies in ignoring them, and the true promise that they hold for our specialties. PMID- 19806090 TI - American College of Gastroenterology Guidelines for Colorectal Cancer Screening 2008. PMID- 19806085 TI - A causal association between isotretinoin and inflammatory bowel disease has yet to be established. AB - OBJECTIVES: Case reports have described a possible association between isotretinoin and inflammatory bowel disease (IBD). We critically appraised the literature on this association to assess whether it supports a causal relationship. METHODS: We systematically searched for case reports, case series, and clinical trials assessing this association. We then applied the Hill criteria to evaluate causality. RESULTS: Twelve case reports and one case series reported an association between isotretinoin use and subsequent development of IBD. Cases occurred in seven countries over a 23-year period and differed with respect to reported isotretinoin dose, duration of treatment before development of disease, whether disease developed on or off medication, and clinical presentation of disease. No prospective or retrospective studies have examined the relationship between isotretinoin and IBD. An estimated 59 coincident cases of IBD would be expected in isotretinoin users each year, assuming no increased risk. Alternative explanations may account for the sequence of events seen in case reports. Strength, specificity, and consistency of the association are lacking. CONCLUSIONS: Current evidence is insufficient to confirm or refute a causal association between isotretinoin and IBD. Additional prospective or well-designed retrospective (e.g., case-control) pharmacoepidemiological studies are needed to definitively establish causality. PMID- 19806091 TI - Colorectal cancer guidance is self-serving. PMID- 19806092 TI - Are split bowel preparation regimens practical for morning colonoscopies? Implications of the new american college of gastroenterology colon cancer screening guidelines for real-world clinical practice. PMID- 19806094 TI - Endoscopy timing in acute variceal hemorrhage: perhaps not the sooner the better, but delay not justified. PMID- 19806096 TI - Radiofrequency ablation and dilated columnar-lined esophagus. PMID- 19806097 TI - Claim for screening endoscopy of the esophagus. PMID- 19806099 TI - Laparoscopic vs. colonoscopic removal of a large polyp. PMID- 19806101 TI - Infections in flares of inflammatory bowel disease. PMID- 19806102 TI - Clostrium cadaveris intra-peritoneal abscess. PMID- 19806103 TI - You cannot see the wood for the tree. PMID- 19806104 TI - Management of biliobronchial fistula with octreotide: a case report. PMID- 19806105 TI - Multifocal gastrointestinal leiomyomas. PMID- 19806106 TI - Splenic hypofunction in Whipple's disease. PMID- 19806107 TI - Rizatriptan induced acute on top of chronic ischemic colitis. PMID- 19806108 TI - When the left got left behind. PMID- 19806109 TI - Use of proton pump inhibitors and risk of progression of Barrett's esophagus to neoplastic lesions. PMID- 19806110 TI - Idiopathic gastric acid hypersecretion in a patient with MUTYH-associated polyposis. PMID- 19806111 TI - Single immunoglobulin infusion can reverse hemodynamic failure associated with severe Clostridium difficile colitis. PMID- 19806112 TI - Isolated pancreatic tuberculosis in a patient with total situs inversus. PMID- 19806113 TI - Case of fulminant colitis after docetaxel therapy for breast carcinoma in a patient with underlying ulcerative colitis. PMID- 19806114 TI - Crohn's disease presenting as acute hypokalemic paralysis. PMID- 19806144 TI - Epidemiology of pancreatic cancer: an overview. AB - Pancreatic cancer, although infrequent, has an exceptionally high mortality rate, making it one of the four or five most common causes of cancer mortality in developed countries. The incidence of pancreatic cancer varies greatly across regions, which suggests roles for lifestyle factors, such as diet, or environmental factors, such as vitamin D exposure. Smoking is the most common known risk factor, and is the cause of 20-25% of all pancreatic tumors. Alcohol does not seem to be a risk factor, unless it leads to chronic pancreatitis, which is a probable risk factor. Long-standing diabetes increases the risk of pancreatic cancer, but can also be an early manifestation of pancreatic tumors. 5 10% of patients with pancreatic cancer have an underlying germline disorder, while the remaining percentage of cancer cases is thought to be caused by somatic mutations. Some individual studies suggest that mutations in various polymorphic genes can lead to small increases in the risk of pancreatic cancer, but these findings need to be replicated. Rising prevalence of smoking in developing countries, improved diagnosis and increasing population longevity are all likely to increase the global burden of pancreatic cancer in the coming decades. PMID- 19806145 TI - New advances that enable identification of glioblastoma recurrence. AB - Postoperative adjuvant radiation therapy and temozolomide chemotherapy have become the standard care for newly diagnosed malignant gliomas. The efficacy of these therapies has led to an increase in pseudoprogression and radiation necrosis, both of which are treatment-related effects whose appearance on standard MRI with gadolinium-based contrast agents resembles that of tumor progression or recurrence. Accurate diagnosis of these post-treatment lesions as either tumor recurrence or treatment effects (pseudoprogression or radiation necrosis) is important to determine the patient's prognosis. Modern advancements with magnetic resonance spectroscopy (MRS), diffusion-weighted imaging (DWI), and PET scans have shown promise for distinguishing tumor recurrence from treatment effects. Advances in radiographic techniques will become critically important with the emergence of new antiangiogenic therapies. Consequently, MRS, DWI, and PET need to be incorporated into routine post-treatment investigations to improve the specificity and sensitivity of distinguishing tumor recurrence from treatment effects. Further research will also be needed to develop improved algorithms that use these modalities, and to develop new modalities with even greater accuracy than those currently available. PMID- 19806146 TI - Myeloproliferative neoplasms: contemporary diagnosis using histology and genetics. AB - The 2008 WHO classification system for hematological malignancies is comprehensive and includes histology and genetic information. Myeloid neoplasms are now classified into five categories: acute myeloid leukemia, myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPN), MDS/MPN, and myeloid and/or lymphoid malignancies associated with eosinophilia and PDGFR or FGFR1 rearrangements. MPN are subclassified into eight separate entities: chronic myelogenous leukemia, polycythemia vera, essential thrombocythemia, primary myelofibrosis, systemic mastocytosis, chronic eosinophilic leukemia not otherwise specified, chronic neutrophilic leukemia, and unclassifiable MPN. The diagnosis of chronic myelogenous leukemia requires the presence of BCR-ABL1, while its absence is required for all other MPN. Additional MPN-associated molecular markers include mutations of JAK2, MPL, TET2 and KIT. JAK2 V617F is found in most patients with polycythemia vera, essential thrombocythemia, or primary myelofibrosis and is, therefore, useful as a clonal marker in those settings. The diagnostic utility of MPL and TET2 mutations is limited by low mutational frequency. In systemic mastocytosis, presence of KIT D816V is expected but not essential for diagnosis. Chronic eosinophilic leukemia not otherwise specified should be distinguished from both PDGFR-rearranged or FGFR1-rearranged neoplasms and hypereosinophilic syndrome. We discuss histologic, cytogenetic and molecular changes in MPN and illustrate their integration into practical diagnostic algorithms. PMID- 19806148 TI - Association of SLC6A4 variants with obsessive-compulsive disorder in a large multicenter US family study. AB - Genetic association studies of SLC6A4 (SERT) and obsessive-compulsive disorder (OCD) have been equivocal. We genotyped 1241 individuals in 278 pedigrees from the OCD Collaborative Genetics Study for 13 single-nucleotide polymorphisms, for the linked polymorphic region (LPR) indel with molecular haplotypes at rs25531, for VNTR polymorphisms in introns 2 and 7 and for a 381-bp deletion 3' to the LPR. We analyzed using the Family-Based Association Test (FBAT) under additive, dominant, recessive and genotypic models, using both OCD and sex-stratified OCD as phenotypes. Two-point FBAT analysis detected association between Int2 (P = 0.0089) and Int7 (P = 0.0187) (genotypic model). Sex-stratified two-point analysis showed strong association in females with Int2 (P<0.0002), significant after correction for linkage disequilibrium, and multiple marker and model testing (P(Adj) = 0.0069). The SLC6A4 gene is composed of two haplotype blocks (our data and the HapMap); FBAT whole-marker analysis conducted using this structure was not significant. Several noteworthy nonsignificant results have emerged. Unlike Hu et al., we found no evidence for overtransmission of the LPR L(A) allele (genotype relative risk = 1.11, 95% confidence interval: 0.77-1.60); however, rare individual haplotypes containing L(A) with P<0.05 were observed. Similarly, three individuals (two with OCD/OCPD) carried the rare I425V SLC6A4 variant, but none of them passed it on to their six OCD-affected offspring, suggesting that it is unlikely to be solely responsible for the 'OCD plus syndrome', as reported by Ozaki et al. In conclusion, we found evidence of genetic association at the SLC6A4 locus with OCD. A noteworthy lack of association at the LPR, LPR-rs25531 and rare 425V variants suggests that hypotheses about OCD risk need revision to accommodate these new findings, including a possible gender effect. PMID- 19806147 TI - Chlamydial infection: the link with ocular adnexal lymphomas. AB - Chlamydiae are obligate intracellular bacteria that grow in eukaryotic cells and cause a wide spectrum of diseases. They can establish persistent infections, are mitogenic in vitro, promote polyclonal cell proliferation in vivo and induce resistance to apoptosis in infected cells-properties that might contribute to tumorigenesis. In fact, Chlamydophila psittaci (Cp) has been linked to the development and maintenance of ocular adnexal marginal zone B-cell lymphoma (OAMZL). In this indolent malignancy, Cp is transported by monocytes and macrophages and causes both local and systemic infection. Cp elementary bodies are viable and infectious in the conjunctiva and peripheral blood of patients with OAMZL. Bacterial eradication with antibiotic therapy is often followed by lymphoma regression. Despite recent advances in the understanding of this bacterium-lymphoma association, several questions remain unanswered. For instance, prevalence variations among different geographical areas and related diagnostic and therapeutic implications remain a major investigational issue. We will focus on clinical and therapeutic implications of chlamydial infections in patients with lymphomas and summarize the current knowledge on the association between Cp infection and OAMZL. Available data on the epidemiology, biology and pathogenesis of this association are analyzed and new investigative and clinical approaches are discussed. PMID- 19806149 TI - Unanticipated demyelinating pathology of the CNS. AB - Incidentally identified anomalies within the CNS that are highly suggestive of demyelinating disease have been extensively described in neuropathological series, and are increasingly being detected during premortem investigations. With the exception of studies that focused specifically on the prevalence of this entity, the observed anomalies are unanticipated and unrelated to the intended purpose of the examination. The discovery of MRI technology, and its subsequent widespread adoption in the clinic, has facilitated the identification of such cases. The natural course of individuals with incidentally identified demyelinating anomalies is unknown at present. This Review focuses on the history and nosology of unexpected demyelinating pathology, encompassing both autopsy data and MRI-based antemortem investigations of large cohorts, as well as family members at high risk of developing multiple sclerosis. Longitudinal clinical data acquired from prospectively followed cohorts will also be reviewed. In addition, I discuss the estimated prevalence of demyelinating pathology, the currently proposed criteria for its identification, implications for therapeutic intervention, and predictors of disease progression. PMID- 19806150 TI - The diagnosis and treatment of early psoriatic arthritis. AB - Psoriatic arthritis (PsA) is a chronic inflammatory musculoskeletal disorder associated with a heterogeneous disease presentation, varied disease expression and an unpredictable but often chronically destructive clinical course. Joint damage can occur early in the disease; indeed, several imaging modalities have demonstrated subclinical joint involvement in psoriasis patients without musculoskeletal signs or symptoms. Efforts are underway to validate questionnaires that will enable dermatologists to screen patients with psoriasis for the presence of musculoskeletal disease. To date, the use of therapies in patients with early PsA has not been reported in randomized controlled trials. Moreover, conventional agents are partially effective in established PsA but, in general, trials with DMARDS have not included validated outcome measures for the different manifestations of PsA. Tumor necrosis factor antagonists can alleviate the signs and symptoms of established psoriatic arthritis and inhibit radiographic progression, but the therapeutic impact of early intervention with these agents requires further study. The extent of disease and the presence of comorbidities should be used to guide treatment decisions and to minimize adverse events. PMID- 19806152 TI - Tools for monitoring spondyloarthritis in clinical practice. AB - Spondyloarthritis (SpA) usually follows a chronic disease course that requires regular medical care and monitoring to control for increased disease activity and to maintain physical function. This Review describes the instruments and imaging techniques available for monitoring SpA in clinical practice. Specifically, questionnaires, physical examination and laboratory tests that can be used to measure physical function, disease activity and pain are discussed. Furthermore, the value of different imaging techniques, including conventional radiography, MRI and ultrasonography, for use in the clinic is described. In the past decade, most of the work on standardizing the monitoring of outcome measures in SpA has focused on ankylosing spondylitis, and similar developments are currently in progress for the monitoring of psoriatic arthritis. This Review, therefore, will focus on these disorders. PMID- 19806151 TI - Oligoarticular and polyarticular JIA: epidemiology and pathogenesis. AB - Juvenile idiopathic arthritis (JIA) refers to a group of chronic childhood arthropathies of unknown etiology, currently classified into subtypes primarily on the basis of clinical features. Research has focused on the hypothesis that these subtypes arise through distinct etiologic pathways. In this Review, we discuss four subtypes of JIA: persistent oligoarticular, extended oligoarticular, rheumatoid-factor-positive polyarticular and rheumatoid-factor-negative polyarticular. These subtypes differ in prevalence between ethnic groups and are associated with different HLA alleles. Non-HLA genetic risk factors have also been identified, some of which reveal further molecular differences between these subtypes, while others suggest mechanistic overlap. Investigations of immunophenotypes also provide insights into subtype differences: adaptive immunity seems to have a prominent role in both polyarticular and oligoarticular JIA, and the more-limited arthritis observed in persistent oligoarticular JIA as compared with extended oligoarticular JIA may reflect more-potent immunoregulatory T-cell activity in the former. Tumor necrosis factor seems to be a key mediator of both polyarticular and oligoarticular JIA, especially in the extended oligoarticular subtype, although elevated levels of other cytokines are also observed. Limited data on monocytes, dendritic cells, B cells, natural killer T cells and neutrophils suggest that the contributions of these cells differ across subtypes of JIA. Within each subtype, however, common pathways seem to drive joint damage. PMID- 19806153 TI - The evolutionary conundrum of pathogen mimicry. AB - Evolutionary conflicts involving mimicry are found throughout nature. Diverse pathogens produce a range of 'mimics' that resemble host components in both form and function. Such mimics subvert crucial cellular processes, including the cell cycle, apoptosis, cytoskeletal dynamics and immunity. Here, we review the mounting evidence that mimicry of host processes is a highly successful strategy for pathogens. Discriminating mimics can be crucial for host survival, and host factors exist that effectively counteract mimics, using strategies that combine rapid evolution and an unexpected degree of flexibility in protein-protein interactions. Even in these instances, mimicry may alter the evolutionary course of fundamental cellular processes in host organisms. PMID- 19806156 TI - Leptin is essential in maintaining normal vascular compliance independent of body weight. AB - The adipocytokine leptin centrally regulates body weight by enhancing metabolic rate and signaling satiety, but it also has wide-ranging peripheral effects. Leptin receptors are expressed on vascular smooth muscle cells and have a role in maintaining vascular tone. We investigated the vascular effects of leptin repletion or calorie restriction on leptin-deficient mice (ob/ob) and a leptin antagonist on wild-type (WT) mice. Aortic compliance was assessed by the measurement of pulse wave velocity by noninvasive Doppler; blood pressure was measured by left ventricular catheterization. We found that ob/ob mice have much stiffer aortas than WT mice and that reduction in aortic stiffness was greater in ob/ob mice treated with leptin vs calorie restriction, despite similar weight loss. Interestingly, treating WT mice with a leptin antagonist increases aortic stiffness with no change in weight. Thus, we conclude that leptin is essential for maintaining normal aortic compliance independent of body weight. PMID- 19806155 TI - Metabolism, cell growth and the bacterial cell cycle. AB - Adaptation to fluctuations in nutrient availability is a fact of life for single celled organisms in the 'wild'. A decade ago our understanding of how bacteria adjust cell cycle parameters to accommodate changes in nutrient availability stemmed almost entirely from elegant physiological studies completed in the 1960s. In this Opinion article we summarize recent groundbreaking work in this area and discuss potential mechanisms by which nutrient availability and metabolic status are coordinated with cell growth, chromosome replication and cell division. PMID- 19806157 TI - Microvascular dysfunction: a direct link among BMI, waist circumference and glucose homeostasis in young overweight/obese normoglycemic women? AB - OBJECTIVE: Capillary recruitment is impaired in obesity (OB), possibly worsening glucose and insulin availability to target organs. In this study, we investigated whether functional microvascular parameters were correlated with clinical anthropometrical data and whether these parameters would influence OB-related metabolic disorders, especially glucose homeostasis, in young overweight (OW)/obese women. DESIGN: Cross-sectional clinical study of microvascular reactivity in young OW/obese women. SUBJECTS AND METHODS: A total of 10 lean (23.1 + or - 3.2 years, body mass index (BMI) 22.3 + or - 1.6 kg m(-2)) and 42 OW/obese (24.9 + or - 3.5 years; BMI 34.5 + or - 5.7 (25.7-46.5) kg m(-2)) sedentary non-smoking women were evaluated. Lipid profile, fasting plasma glucose (PG), post-load PG (75 g-2 h), insulin, C-reactive protein, HOMA-IR (homeostasis model assessment for insulin resistance) index and anthropometric variables (weight, BMI, waist and hip circumferences, waist-to-hip ratio and blood pressure (BP)) were determined. Functional microvascular parameters (functional capillary density, red blood cell velocity at baseline and peak (RBCV(max)), and time taken to reach RBCV(max) (TRBCV(max)) during post-occlusive reactive hyperemia after 1 min arterial occlusion) were evaluated by nailfold videocapillaroscopy. RESULTS: The time taken to reach RBCV(max) was significantly longer in OW/obese patients compared with control subjects (8.6 + or - 2.4 versus 5.7 + or - 1.1 s, P<0.001), and its delay was directly associated with adiposity levels, systolic BP and insulin resistance, and inversely related to high-density lipoprotein cholesterol. Post-load PG could be correlated with TRBCV(max) (R = 0.48, P<0.05) and RBCV(max) (R = -0.29, P<0.05), and it was influenced by weight, waist circumference and TRBCV(max) (adjusted R(2) = 24%) as well. CONCLUSIONS: In the investigated group of young OW/obese women, the direct correlation between post load PG and TRBCV(max) links microvascular parameters with metabolic variables and suggests a key role for microcirculation in OB-related metabolic disorders. PMID- 19806154 TI - The trypanosome flagellar pocket. AB - Trypanosomes are important disease agents and excellent models for the study of evolutionary cell biology. The trypanosome flagellar pocket is a small invagination of the plasma membrane where the flagellum exits the cytoplasm and participates in many cellular processes. It is the only site of exocytosis and endocytosis and part of a multiorganelle complex that is involved in cell polarity and cell division. Several flagellar pocket-associated proteins have been identified and found to contribute to trafficking and virulence. In this Review we discuss the contribution of the flagellar pocket to protein trafficking, immune evasion and other processes. PMID- 19806158 TI - Increased dorsolateral prefrontal cortex activation in obese children during observation of food stimuli. AB - OBJECTIVE: Food cues yield different patterns of brain activation in obese compared with normal-weight adults in prefrontal and limbic/paralimbic areas. For children, no mapping studies comparing representation sites for food and other stimuli between obese and normal-weight subjects are available. DESIGN: We used a cross-sectional design of two age-matched subject groups to investigate differences in brain activation in response to visually presented food, pleasant, and neutral pictures between obese/overweight and normal children. SUBJECTS: 22 overweight/obese children were compared with 22 normal-weight children. MEASUREMENTS: Functional magnetic resonance imaging (of the whole head during perception of visually presented stimuli), psychological testing, and psychophysical measures of heart rate deceleration were assessed. RESULTS: Obese children showed higher activation of the dorsolateral prefrontal cortex (DLPFC) in response to food pictures. In addition, DLPFC activation was negatively correlated with self-esteem. In contrast, normal-weight children showed higher activation of the caudate and hippocampus specific to food pictures, and of the anterior cingulate cortex and thalamus to visual cues in general. In response to food stimuli, obese children showed a heart rate deceleration correlating positively with activation of the ventrolateral prefrontal cortex. CONCLUSION: Obese children react to food stimuli with increased prefrontal activation, which might be associated with increased inhibitory control. PMID- 19806159 TI - High fat stores in ectopic compartments in men with newly diagnosed type 2 diabetes: an anthropometric determinant of carotid atherosclerosis and insulin resistance. AB - OBJECTIVE: The objective of this study was to analyze how fat partitioning in the liver, muscle and visceral compartments is altered by diabetes and age, and whether altered fat distribution is associated with a higher carotid artery intima-media thickness (C-IMT) and insulin resistance. METHODS: This was an observational study performed on 21 young healthy men (mean age + or - s.d., 28.6 + or - 3.8 years) and 73 men with newly developed type 2 diabetes (38 young (29.2 + or - 4.1 years) and 35 middle-aged (47.1 + or - 6.0 years) subjects). Abdominal visceral and subcutaneous fat areas, mid-thigh muscle attenuation and liver attenuation characteristics were determined; the mid-thigh muscle was divided into low- and normal-density muscle areas. RESULTS: The young and middle-aged diabetic subjects had higher visceral fat areas, higher liver attenuation and higher lipid-rich muscle (greater low-density muscle area and decreased muscle attenuation) when compared with healthy individuals; however, no differences were observed between the two diabetic groups. In contrast, the C-IMT increased with both age and diabetes. On the basis of multiple regression analyses, mid-thigh low-density muscle area and muscle attenuation were independently associated with the C-IMT, and the mid-thigh normal-density muscle area and muscle attenuation were independent factors of insulin resistance. CONCLUSIONS: High fat stores within ectopic compartments were observed at an early stage in the development of diabetes. Furthermore, altered lipid partitioning within muscle was independently associated with carotid atherosclerosis and insulin resistance. PMID- 19806160 TI - Joint associations of physical activity and sedentary behaviors with body mass index: results from a time use survey of US adults. AB - OBJECTIVE: Obesity risk is negatively associated with physical activity and positively associated with time spent in sedentary behaviors. Yet, it is not known how different combinations of sedentary and active behavior are associated with body mass index (BMI). This study examined the interaction between time spent in physical activity and sedentary behavior on BMI in US adults. DESIGN: Cross-sectional, data from the 2006 American Time Use Survey. SUBJECTS: 10 984 non-underweight adults (aged 21 + years). MEASUREMENT: A phone interview assessed all activities performed in the past 24 h, height, weight, health status, and other sociodemographic characteristics. Time spent in (1) moderate-to-vigorous leisure-time physical activity (MVPA), (2) active transportation (walking, biking), (3) sedentary leisure activities (TV/movie watching, computer use, playing games, reading), and (4) sedentary transportation (motorized vehicles) was determined from activity coding. BMI was calculated. RESULTS: After adjusting for age, gender, education level, race/ethnicity, and health status, sample weighted linear regressions found significant interactions for leisure MVPA x TV/movies, leisure MVPA x playing games, active transportation x sedentary transportation, and active transportation x reading (Ps<0.0001). For example, the group of adults watching <60 min per day of TV/movies and engaging in > or =60 min per day of leisure MVPA had lower average BMI compared to the group watching <60 min per day of TV/movies and reporting <60 min per day of leisure MVPA (P<0.0001). In contrast, for adults watching > or =189 min per day of TV/movies, there was not a significant difference in BMI by time spent in leisure MVPA. CONCLUSION: Data from a US time use survey indicate that the strength of the association between certain types of sedentary behavior and BMI varies according to time spent in certain types of physical activity and vice versa. PMID- 19806161 TI - Epidemiological profile of a pediatric population with acquired spinal cord injury from AACD: Sao Paulo/Brazil. AB - OBJECTIVE: To provide a retrospective analysis of pediatric patients with Spinal Cord Injury (SCI), who were admitted for treatment and rehabilitation in a reference Brazilian medical center during the past 6 years. METHODS: This retrospective analysis included 106 patients 16 years of age or younger who were admitted with SCI in our Rehabilitation Medical Center between April 2002 and June 2008. The patients admitted were analyzed according to their gender, age, origin region, cause and level of injury and clinical complications. RESULTS/CONCLUSIONS: The frequency corresponded to 5.4% of the total of SCI cases studied. The 106 patients reviewed included 67 boys (63.2%) and the mean age of 8.6 years. In all, 50.9% of the SCI have traumatic etiology. The most frequent causes were gunshot 42.6%, traffic accident 38.9%, diving 9.3% and fall 3.7%. The nontraumatic etiologies corresponded to 49.1% of the total patients. Overall, tumor (36.5%) and infection (19.2%) were the most frequent cause of spinal injuries. The average time between SCI event and arrival at the rehabilitation medical center was 27 months. The majority of patients were paraplegic (76.4%). In the first assessment 64.2% patients showed clinical complications, such as orthopedic deformity (69.1%), pressure ulcers (35.3%), spasticity (8.8%), obesity (2.9%). Pediatric SCI is a rare but devastating condition affecting the patient, patient's family and society. An understanding of the etiology and the frequency of pediatric SCI can improve prevention and treatment. PMID- 19806162 TI - Skimboarding: a new cause of water sport spinal cord injury. AB - STUDY DESIGN: Case series. OBJECTIVE: To present three cases of spinal cord injuries associated with skimboarding and to suggest aspects of the sport that may be associated with spinal cord injury. SETTING: Shepherd Center, Atlanta, GA, USA. Spinal cord injury rehabilitation facility. METHODS: Three males, aged 17-23 years, sustained cervical spine fractures resulting in tetraplegia after skimboarding accidents. RESULTS: The patients admitted from Florida hospitals presented with tetraplegia resulting from both incomplete and complete spinal cord injuries. The ASIA Impairment Scale (AIS) neurologic levels ranged from C3 to C5. All were injured by being thrown from the skimboard riding into the waves in shallow water. All patients required surgical stabilization and inpatient rehabilitation. CONCLUSIONS: Previous studies on skimboarding have demonstrated the risk of extremity fractures and soft tissue injuries. Spinal cord injury should be considered an additional risk associated with skimboarding, particularly as the sport has grown in popularity and become more 'extreme' in the maneuvers performed. Increased education and awareness about the potential risk of spinal cord injury are essential. PMID- 19806163 TI - Narrowing of T-cell receptor beta variable repertoire during symptomatic herpesvirus infection in transplant patients. AB - Primary infection or recrudescence of latent virus infection in transplant recipients can be manifested either as asymptomatic or symptomatic disease. Here we show that symptomatic human cytomegalovirus (HCMV) or Epstein-Barr virus (EBV) infection or recrudescence following solid organ transplantation (SOT) was coincident with a dramatic skewing of T-cell receptor beta variable (TRBV) repertoire, with expansions of monoclonal/oligoclonal clonotypes. As the clinical symptoms resolved, the peripheral blood repertoire reverted to a more diverse distribution. In contrast, SOT recipients with asymptomatic or no viral infection or recrudescence showed minimal or no skewing of the T-cell receptor repertoire to maintain peripheral blood repertoire diversity. More importantly, we show that large monoclonal/oligoclonal repertoire expansions are associated with the loss of HCMV-specific T-cell function observed in SOT patients undergoing symptomatic viral infection or recrudescence, whereas SOT recipients who maintain peripheral blood TRBV repertoire diversity and functional antigen-specific T-cell responses can resist clinical symptomatic disease in spite of high levels of viral load. PMID- 19806164 TI - The role of notch signaling in the development of a normal B-cell repertoire. AB - The notch signaling pathway is evolutionarily conserved across the animal kingdom and regulates developmental 'decisions', such as cell fate commitment, differentiation, proliferation and apoptosis. In the mammalian immune system, notch signaling events have been extensively studied during T lymphopoiesis, and have a role both during early development, as well as differentiation into discreet effector cell compartments. In contrast, the impact of notch signaling in the B-cell compartment is less obvious. It is clear that notch signaling is crucial to generate the marginal zone B-cell population located within the spleen; however, the full effects of notch signaling during normal B-cell development remain unresolved. Nevertheless, there is compelling evidence that notch signaling regulates multiple stages of B-cell differentiation and in shaping the antibody repertoire; however, the molecular details have not been elucidated. This review explores the relationship between notch signaling and B cell development with attention to how these processes contribute to a normal B cell repertoire. PMID- 19806165 TI - Inhibition of cytokinesis by overexpression of NudCL that is localized to the centrosome and midbody. PMID- 19806166 TI - The Arabidopsis PARAQUAT RESISTANT2 gene encodes an S-nitrosoglutathione reductase that is a key regulator of cell death. AB - Metabolism of S-nitrosoglutathione (GSNO), a major biologically active nitric oxide (NO) species, is catalyzed by the evolutionally conserved GSNO reductase (GSNOR). Previous studies showed that the Arabidopsis GSNOR1/HOT5 gene regulates salicylic acid signaling and thermotolerance by modulating the intracellular S nitrosothiol level. Here, we report the characterization of the Arabidopsis paraquat resistant2-1 (par2-1) mutant that shows an anti-cell death phenotype. The production of superoxide in par2-1 is comparable to that of wild-type plants when treated by paraquat (1,1'-dimethyl-4,4'-bipyridinium dichloride), suggesting that PAR2 acts downstream of superoxide to regulate cell death. PAR2, identified by positional cloning, is shown to be identical to GSNOR1/HOT5. The par2-1 mutant carries a missense mutation in a highly conserved glycine, which renders the mutant protein unstable. Compared to wild type, par2-1 mutant has a higher NO level, as revealed by staining with 4,5-diaminofluorescein diacetate. Consistent with this result, wild-type plants treated with an NO donor display resistance to paraquat. Interestingly, the GSNOR1/HOT5/PAR2 protein level, other than its steady-state mRNA level, is induced by paraquat, but is reduced by NO donors. Taken together, these results suggest that GSNOR1/HOT5/PAR2 plays an important role in regulating cell death in plant cells through modulating intracellular NO level. PMID- 19806167 TI - Urinary incontinence: Relative efficacy of retropubic and transobturator slings. PMID- 19806168 TI - Health policy: Do end-of-life discussions have a role in health care reform? PMID- 19806169 TI - Stones: Impact of dose reduction on CT detection of urolithiasis. PMID- 19806170 TI - Prostate cancer: Optimizing the duration of androgen deprivation therapy. PMID- 19806171 TI - Prostate cancer: How can we improve the health of men who receive ADT? PMID- 19806172 TI - Prostatitis: Clinical phenotyping of patients with pelvic pain. PMID- 19806173 TI - Bilateral renal hemorrhage due to polyarteritis nodosa wrongly attributed to blunt trauma. AB - BACKGROUND: A 36-year-old, previously healthy man presented to an accident and emergency department with right-sided abdominal pain 7 days after he sustained a trauma to his right flank. He was using no medication other than over-the-counter analgesics since his injury. INVESTIGATIONS: Complete blood count, serum creatinine measurement, liver function tests, hepatitis B and C screening, abdominal CT, renal angiography, surgical exploration and histology of kidney samples. DIAGNOSIS: Polyarteritis nodosa with Page kidney causing bilateral perirenal hematoma, severe hypertension and renal failure. MANAGEMENT: The patient was severely anemic, and his bleeding was investigated. A 15 x 13 x 12 cm retroperitoneal hematoma was found in the region of the right kidney and the patient underwent unilateral right nephrectomy. 3 weeks after discharge the patient was readmitted with a left-sided perirenal hematoma. Steel-coil embolization of the kidney stopped the bleeding but the patient developed hypertension and renal failure, and antihypertensive agents and dialysis were started. Microaneurysms and vessel-wall necrosis were discovered on re examination of the angiogram and histology, respectively, so immunosuppressive therapy was started, comprising intravenous methylprednisolone daily for 3 days and oral prednisolone and intravenous cyclophosphamide for 4 weeks. Page kidney, resulting from the bleeding into the solitary kidney, caused stretching of the renal artery and deterioration of renal function, which required hemodialysis treatment. PMID- 19806174 TI - A rare case of renal infarction caused by infective endocarditis. AB - BACKGROUND: A 29-year-old man presented to the emergency department of a general hospital complaining of sudden onset left loin pain, radiating to the groin, which had started 48 h previously. He described no urological symptoms and had no medical history of note. INVESTIGATIONS: Physical examination, electrocardiography, dipstick testing of urine, radiography of the chest and abdomen, blood tests (white blood cell count and serum urea, creatinine, sodium, potassium and C-reactive protein levels), CT of the renal tract, blood and urine cultures, renal angiography, thromboembolic blood panel, urine and blood tests for illicit drugs, transthoracic echocardiography, transesophageal echocardiography, renal ultrasonography. DIAGNOSIS: Infective endocarditis resulting in thromboembolic unilateral renal infarction. MANAGEMENT: The patient was started on anticoagulation therapy with low-molecular-weight heparin and treated with intravenous gentamicin and benzylpenicillin for 4 weeks. He was seen in an outpatient clinic 4 weeks after discharge, at which time serum urea and creatinine levels and repeat ultrasonography of the renal tract confirmed normal renal function. He will be followed up regularly by cardiologists and urologists, at 6 weeks initially, and every 6 months to 1 year thereafter by his family physician. PMID- 19806175 TI - Genomic diversity and evolution of Mycobacterium ulcerans revealed by next generation sequencing. AB - Mycobacterium ulcerans is the causative agent of Buruli ulcer, the third most common mycobacterial disease after tuberculosis and leprosy. It is an emerging infectious disease that afflicts mainly children and youths in West Africa. Little is known about the evolution and transmission mode of M. ulcerans, partially due to the lack of known genetic polymorphisms among isolates, limiting the application of genetic epidemiology. To systematically profile single nucleotide polymorphisms (SNPs), we sequenced the genomes of three M. ulcerans strains using 454 and Solexa technologies. Comparison with the reference genome of the Ghanaian classical lineage isolate Agy99 revealed 26,564 SNPs in a Japanese strain representing the ancestral lineage. Only 173 SNPs were found when comparing Agy99 with two other Ghanaian isolates, which belong to the two other types previously distinguished in Ghana by variable number tandem repeat typing. We further analyzed a collection of Ghanaian strains using the SNPs discovered. With 68 SNP loci, we were able to differentiate 54 strains into 13 distinct SNP haplotypes. The average SNP nucleotide diversity was low (average 0.06-0.09 across 68 SNP loci), and 96% of the SNP locus pairs were in complete linkage disequilibrium. We estimated that the divergence of the M. ulcerans Ghanaian clade from the Japanese strain occurred 394 to 529 thousand years ago. The Ghanaian subtypes diverged about 1000 to 3000 years ago, or even much more recently, because we found evidence that they evolved significantly faster than average. Our results offer significant insight into the evolution of M. ulcerans and provide a comprehensive report on genetic diversity within a highly clonal M. ulcerans population from a Buruli ulcer endemic region, which can facilitate further epidemiological studies of this pathogen through the development of high resolution tools. PMID- 19806176 TI - SnTox3 acts in effector triggered susceptibility to induce disease on wheat carrying the Snn3 gene. AB - The necrotrophic fungus Stagonospora nodorum produces multiple proteinaceous host selective toxins (HSTs) which act in effector triggered susceptibility. Here, we report the molecular cloning and functional characterization of the SnTox3 encoding gene, designated SnTox3, as well as the initial characterization of the SnTox3 protein. SnTox3 is a 693 bp intron-free gene with little obvious homology to other known genes. The predicted immature SnTox3 protein is 25.8 kDa in size. A 20 amino acid signal sequence as well as a possible pro sequence are predicted. Six cysteine residues are predicted to form disulfide bonds and are shown to be important for SnTox3 activity. Using heterologous expression in Pichia pastoris and transformation into an avirulent S. nodorum isolate, we show that SnTox3 encodes the SnTox3 protein and that SnTox3 interacts with the wheat susceptibility gene Snn3. In addition, the avirulent S. nodorum isolate transformed with SnTox3 was virulent on host lines expressing the Snn3 gene. SnTox3-disrupted mutants were deficient in the production of SnTox3 and avirulent on the Snn3 differential wheat line BG220. An analysis of genetic diversity revealed that SnTox3 is present in 60.1% of a worldwide collection of 923 isolates and occurs as eleven nucleotide haplotypes resulting in four amino acid haplotypes. The cloning of SnTox3 provides a fundamental tool for the investigation of the S. nodorum-wheat interaction, as well as vital information for the general characterization of necrotroph-plant interactions. PMID- 19806177 TI - Mucosal immunization with iron receptor antigens protects against urinary tract infection. AB - Uncomplicated infections of the urinary tract, caused by uropathogenic Escherichia coli, are among the most common diseases requiring medical intervention. A preventive vaccine to reduce the morbidity and fiscal burden these infections have upon the healthcare system would be beneficial. Here, we describe the results of a large-scale selection process that incorporates bioinformatic, genomic, transcriptomic, and proteomic screens to identify six vaccine candidates from the 5379 predicted proteins encoded by uropathogenic E. coli strain CFT073. The vaccine candidates, ChuA, Hma, Iha, IreA, IroN, and IutA, all belong to a functional class of molecules that is involved in iron acquisition, a process critical for pathogenesis in all microbes. Intranasal immunization of CBA/J mice with these outer membrane iron receptors elicited a systemic and mucosal immune response that included the production of antigen specific IgM, IgG, and IgA antibodies. The cellular response to vaccination was characterized by the induction and secretion of IFN-gamma and IL-17. Of the six potential vaccine candidates, IreA, Hma, and IutA provided significant protection from experimental infection. In immunized animals, class-switching from IgM to IgG and production of antigen-specific IgA in the urine represent immunological correlates of protection from E. coli bladder colonization. These findings are an important first step toward the development of a subunit vaccine to prevent urinary tract infections and demonstrate how targeting an entire class of molecules that are collectively required for pathogenesis may represent a fundamental strategy to combat infections. PMID- 19806178 TI - Differential regulation of type I interferon and epidermal growth factor pathways by a human Respirovirus virulence factor. AB - A number of paramyxoviruses are responsible for acute respiratory infections in children, elderly and immuno-compromised individuals, resulting in airway inflammation and exacerbation of chronic diseases like asthma. To understand the molecular pathogenesis of these infections, we searched for cellular targets of the virulence protein C of human parainfluenza virus type 3 (hPIV3-C). We found that hPIV3-C interacts directly through its C-terminal domain with STAT1 and GRB2, whereas C proteins from measles or Nipah viruses failed to do so. Binding to STAT1 explains the previously reported capacity of hPIV3-C to block type I interferon signaling, but the interaction with GRB2 was unexpected. This adaptor protein bridges Epidermal Growth Factor (EGF) receptor to MAPK/ERK pathway, a signaling cascade recently found to be involved in airway inflammatory response. We report that either hPIV3 infection or transient expression of hPIV3-C both increase cellular response to EGF, as assessed by Elk1 transactivation and phosphorylation levels of ERK1/2, 40S ribosomal subunit protein S6 and translation initiation factor 4E (eIF4E). Furthermore, inhibition of MAPK/ERK pathway with U0126 prevented viral protein expression in infected cells. Altogether, our data provide molecular basis to explain the role of hPIV3-C as a virulence factor and determinant of pathogenesis and demonstrate that Paramyxoviridae have evolved a single virulence factor to block type I interferon signaling and to boost simultaneous cellular response to growth factors. PMID- 19806180 TI - Packages of care for mental, neurological, and substance use disorders in low- and middle-income countries: PLoS Medicine Series. PMID- 19806179 TI - Packages of care for depression in low- and middle-income countries. PMID- 19806181 TI - Capzb2 interacts with beta-tubulin to regulate growth cone morphology and neurite outgrowth. AB - Capping protein (CP) is a heterodimer that regulates actin assembly by binding to the barbed end of F-actin. In cultured nonneuronal cells, each CP subunit plays a critical role in the organization and dynamics of lamellipodia and filopodia. Mutations in either alpha or beta CP subunit result in retinal degeneration in Drosophila. However, the function of CP subunits in mammalian neurons remains unclear. Here, we investigate the role of the beta CP subunit expressed in the brain, Capzb2, in growth cone morphology and neurite outgrowth. We found that silencing Capzb2 in hippocampal neurons resulted in short neurites and misshapen growth cones in which microtubules overgrew into the periphery and completely overlapped with F-actin. In searching for the mechanisms underlying these cytoskeletal abnormalities, we identified beta-tubulin as a novel binding partner of Capzb2 and demonstrated that Capzb2 decreases the rate and the extent of tubulin polymerization in vitro. We mapped the region of Capzb2 that was required for the subunit to interact with beta-tubulin and inhibit microtubule polymerization. A mutant Capzb2 lacking this region was able to bind F-actin and form a CP heterodimer with alpha2-subunit. However, this mutant was unable to rescue the growth cone and neurite outgrowth phenotypes caused by Capzb2 knockdown. Together, these data suggest that Capzb2 plays an important role in growth cone formation and neurite outgrowth and that the underlying mechanism may involve direct interaction between Capzb2 and microtubules. PMID- 19806182 TI - Kissing G domains of MnmE monitored by X-ray crystallography and pulse electron paramagnetic resonance spectroscopy. AB - MnmE, which is involved in the modification of the wobble position of certain tRNAs, belongs to the expanding class of G proteins activated by nucleotide dependent dimerization (GADs). Previous models suggested the protein to be a multidomain protein whose G domains contact each other in a nucleotide dependent manner. Here we employ a combined approach of X-ray crystallography and pulse electron paramagnetic resonance (EPR) spectroscopy to show that large domain movements are coupled to the G protein cycle of MnmE. The X-ray structures show MnmE to be a constitutive homodimer where the highly mobile G domains face each other in various orientations but are not in close contact as suggested by the GDP-AlF(x) structure of the isolated domains. Distance measurements by pulse double electron-electron resonance (DEER) spectroscopy show that the G domains adopt an open conformation in the nucleotide free/GDP-bound and an open/closed two-state equilibrium in the GTP-bound state, with maximal distance variations of 18 A. With GDP and AlF(x), which mimic the transition state of the phosphoryl transfer reaction, only the closed conformation is observed. Dimerization of the active sites with GDP-AlF(x) requires the presence of specific monovalent cations, thus reflecting the requirements for the GTPase reaction of MnmE. Our results directly demonstrate the nature of the conformational changes MnmE was previously suggested to undergo during its GTPase cycle. They show the nucleotide dependent dynamic movements of the G domains around two swivel positions relative to the rest of the protein, and they are of crucial importance for understanding the mechanistic principles of this GAD. PMID- 19806183 TI - Rational extension of the ribosome biogenesis pathway using network-guided genetics. AB - Biogenesis of ribosomes is an essential cellular process conserved across all eukaryotes and is known to require >170 genes for the assembly, modification, and trafficking of ribosome components through multiple cellular compartments. Despite intensive study, this pathway likely involves many additional genes. Here, we employ network-guided genetics-an approach for associating candidate genes with biological processes that capitalizes on recent advances in functional genomic and proteomic studies-to computationally identify additional ribosomal biogenesis genes. We experimentally evaluated >100 candidate yeast genes in a battery of assays, confirming involvement of at least 15 new genes, including previously uncharacterized genes (YDL063C, YIL091C, YOR287C, YOR006C/TSR3, YOL022C/TSR4). We associate the new genes with specific aspects of ribosomal subunit maturation, ribosomal particle association, and ribosomal subunit nuclear export, and we identify genes specifically required for the processing of 5S, 7S, 20S, 27S, and 35S rRNAs. These results reveal new connections between ribosome biogenesis and mRNA splicing and add >10% new genes-most with human orthologs-to the biogenesis pathway, significantly extending our understanding of a universally conserved eukaryotic process. PMID- 19806184 TI - Randomized controlled trial of RTS,S/AS02D and RTS,S/AS01E malaria candidate vaccines given according to different schedules in Ghanaian children. AB - BACKGROUND: The target delivery channel of RTS,S candidate malaria vaccines in malaria-endemic countries in Africa is the World Health Organisation Expanded Program on Immunization. As an Adjuvant System, age de-escalation and schedule selection step, this study assessed 3 schedules of RTS,S/AS01(E) and RTS,S/AS02(D) in infants and young children 5-17 months of age in Ghana. METHODOLOGY: A Phase II, partially-blind randomized controlled study (blind to vaccine, not to schedule), of 19 months duration was conducted in two (2) centres in Ghana between August 2006 and May 2008. Subjects were allocated randomly (1:1:1:1:1:1) to one of six study groups at each study site, each defining which vaccine should be given and by which schedule (0,1-, 0,1,2- or 0,1,7-months). For the 0,1,2-month schedule participants received RTS,S/AS01(E) or rabies vaccine at one center and RTS,S/AS01(E) or RTS,S/AS02(D) at the other. For the other schedules at both study sites, they received RTS,S/AS01(E) or RTS,S/AS02(D). The primary outcome measure was the occurrence of serious adverse events until 10 months post dose 1. RESULTS: The number of serious adverse events reported across groups was balanced. One child had a simple febrile convulsion, which evolved favourably without sequelae, considered to be related to RTS,S/AS01(E) vaccination. Low grade reactions occurred slightly more frequently in recipients of RTS,S/AS than rabies vaccines; grade 3 reactions were infrequent. Less local reactogenicity occurred with RTS,S/AS01(E) than RTS,S/AS02(D). Both candidate vaccines were highly immunogenic for anti-circumsporozoite and anti-Hepatitis B Virus surface antigen antibodies. Recipients of RTS,S/AS01(E) compared to RTS,S/AS02(D) had higher peak anti-circumsporozoite antibody responses for all 3 schedules. Three dose schedules were more immunogenic than 2 dose schedules. Area under the curve analyses for anti-circumsporozoite antibodies were comparable between the 0,1,2- and 0,1,7-month RTS,S/AS01(E) schedules. CONCLUSIONS: Both candidate malaria vaccines were well tolerated. Anti-circumsporozoite responses were greater with RTS,S/AS01(E) than RTS,S/AS02(D) and when 3 rather than 2 doses were given. This study supports the selection of RTS,S/AS01(E) and a 3 dose schedule for further development in children and infants. TRIAL REGISTRATION: ClinicalTrials.gov NCT00360230. PMID- 19806185 TI - Multi-determinants analysis of molecular alterations for predicting clinical benefit to EGFR-targeted monoclonal antibodies in colorectal cancer. AB - BACKGROUND: KRAS mutations occur in 35-45% of metastatic colorectal cancers (mCRC) and preclude responsiveness to EGFR-targeted therapy with cetuximab or panitumumab. However, less than 20% patients displaying wild-type KRAS tumors achieve objective response. Alterations in other effectors downstream of the EGFR, such as BRAF, and deregulation of the PIK3CA/PTEN pathway have independently been found to give rise to resistance. We present a comprehensive analysis of KRAS, BRAF, PIK3CA mutations, and PTEN expression in mCRC patients treated with cetuximab or panitumumab, with the aim of clarifying the relative contribution of these molecular alterations to resistance. METHODOLOGY/PRINCIPAL FINDINGS: We retrospectively analyzed objective tumor response, progression-free (PFS) and overall survival (OS) together with the mutational status of KRAS, BRAF, PIK3CA and expression of PTEN in 132 tumors from cetuximab or panitumumab treated mCRC patients. Among the 106 non-responsive patients, 74 (70%) had tumors with at least one molecular alteration in the four markers. The probability of response was 51% (22/43) among patients with no alterations, 4% (2/47) among patients with 1 alteration, and 0% (0/24) for patients with > or =2 alterations (p<0.0001). Accordingly, PFS and OS were increasingly worse for patients with tumors harboring none, 1, or > or =2 molecular alteration(s) (p<0.001). CONCLUSIONS/SIGNIFICANCE: When expression of PTEN and mutations of KRAS, BRAF and PIK3CA are concomitantly ascertained, up to 70% of mCRC patients unlikely to respond to anti-EGFR therapies can be identified. We propose to define as 'quadruple negative', the CRCs lacking alterations in KRAS, BRAF, PTEN and PIK3CA. Comprehensive molecular dissection of the EGFR signaling pathways should be considered to select mCRC patients for cetuximab- or panitumumab-based therapies. PMID- 19806186 TI - Functional, non-clonal IgMa-restricted B cell receptor interactions with the HIV 1 envelope gp41 membrane proximal external region. AB - The membrane proximal external region (MPER) of HIV-1 gp41 has several features that make it an attractive antibody-based vaccine target, but eliciting an effective gp41 MPER-specific protective antibody response remains elusive. One fundamental issue is whether the failure to make gp41 MPER-specific broadly neutralizing antibodies like 2F5 and 4E10 is due to structural constraints with the gp41 MPER, or alternatively, if gp41 MPER epitope-specific B cells are lost to immunological tolerance. An equally important question is how B cells interact with, and respond to, the gp41 MPER epitope, including whether they engage this epitope in a non-canonical manner i.e., by non-paratopic recognition via B cell receptors (BCR). To begin understanding how B cells engage the gp41 MPER, we characterized B cell-gp41 MPER interactions in BALB/c and C57BL/6 mice. Surprisingly, we found that a significant (approximately 7%) fraction of splenic B cells from BALB/c, but not C57BL/6 mice, bound the gp41 MPER via their BCRs. This strain-specific binding was concentrated in IgM(hi) subsets, including marginal zone and peritoneal B1 B cells, and correlated with enriched fractions (approximately 15%) of gp41 MPER-specific IgM secreted by in vitro-activated splenic B cells. Analysis of Igh(a) (BALB/c) and Igh(b) (C57BL/6) congenic mice demonstrated that gp41 MPER binding was controlled by determinants of the Igh(a) locus. Mapping of MPER gp41 interactions with IgM(a) identified MPER residues distinct from those to which mAb 2F5 binds and demonstrated the requirement of Fc C(H) regions. Importantly, gp41 MPER ligation produced detectable BCR-proximal signaling events, suggesting that interactions between gp41 MPER and IgM(a) determinants may elicit partial B cell activation. These data suggest that low avidity, non-paratopic interactions between the gp41 MPER and membrane Ig on naive B cells may interfere with or divert bnAb responses. PMID- 19806187 TI - Dendritic cell cross-priming is essential for immune responses to Listeria monocytogenes. AB - Cross-presentation is now recognized as a major mechanism for initiating CD8 T cell responses to virus and tumor antigens in vivo. It provides an elegant mechanism that allows relatively few Dendritic cells (DCs) to initiate primary immune responses while avoiding the consumptive nature of pathogenic infection. CD8 T cells play a major role in anti-bacterial immune responses; however, the contribution of cross-presentation for priming CD8 T cell responses to bacteria, in vivo, is not well established. Listeria monocytogenes (Listeria) is the causative agent of Listeriosis, an opportunistic food-borne bacterial infection that poses a significant public health risk. Here, we employ a transgenic mouse model in which cross-presentation is uniquely inactivated, to investigate cross priming during primary Listeria infection. We show that cross-priming deficient mice are severely compromised in their ability to generate antigen-specific T cells to stimulate MHC I-restricted CTL responses following Listeria infection. The defect in generation of Listeria-elicited CD8 T cell responses is also apparent in vitro. However, in this setting, the endogenous route of processing Listeria-derived antigens is predominant. This reveals a new experimental dichotomy whereby functional sampling of Listeria-derived antigens in vivo but not in vitro is dependent on cross-presentation of exogenously derived antigen. Thus, under normal physiological circumstances, cross-presentation is demonstrated to play an essential role in priming CD8 T cell responses to bacteria. PMID- 19806188 TI - Investigating bacterial sources of toxicity as an environmental contributor to dopaminergic neurodegeneration. AB - Parkinson disease (PD) involves progressive neurodegeneration, including loss of dopamine (DA) neurons from the substantia nigra. Select genes associated with rare familial forms of PD function in cellular pathways, such as the ubiquitin proteasome system (UPS), involved in protein degradation. The misfolding and accumulation of proteins, such as alpha-synuclein, into inclusions termed Lewy Bodies represents a clinical hallmark of PD. Given the predominance of sporadic PD among patient populations, environmental toxins may induce the disease, although their nature is largely unknown. Thus, an unmet challenge surrounds the discovery of causal or contributory neurotoxic factors that could account for the prevalence of sporadic PD. Bacteria within the order Actinomycetales are renowned for their robust production of secondary metabolites and might represent unidentified sources of environmental exposures. Among these, the aerobic genera, Streptomyces, produce natural proteasome inhibitors that block protein degradation and may potentially damage DA neurons. Here we demonstrate that a metabolite produced by a common soil bacterium, S. venezuelae, caused DA neurodegeneration in the nematode, Caenorhabditis elegans, which increased as animals aged. This metabolite, which disrupts UPS function, caused gradual degeneration of all neuronal classes examined, however DA neurons were particularly vulnerable to exposure. The presence of DA exacerbated toxicity because neurodegeneration was attenuated in mutant nematodes depleted for tyrosine hydroxylase (TH), the rate-limiting enzyme in DA production. Strikingly, this factor caused dose-dependent death of human SH-SY5Y neuroblastoma cells, a dopaminergic line. Efforts to purify the toxic activity revealed that it is a highly stable, lipophilic, and chemically unique small molecule. Evidence of a robust neurotoxic factor that selectively impacts neuronal survival in a progressive yet moderate manner is consistent with the etiology of age-associated neurodegenerative diseases. Collectively, these data suggest the potential for exposures to the metabolites of specific common soil bacteria to possibly represent a contributory environmental component to PD. PMID- 19806189 TI - Determining protein complex connectivity using a probabilistic deletion network derived from quantitative proteomics. AB - Protein complexes are key molecular machines executing a variety of essential cellular processes. Despite the availability of genome-wide protein-protein interaction studies, determining the connectivity between proteins within a complex remains a major challenge. Here we demonstrate a method that is able to predict the relationship of proteins within a stable protein complex. We employed a combination of computational approaches and a systematic collection of quantitative proteomics data from wild-type and deletion strain purifications to build a quantitative deletion-interaction network map and subsequently convert the resulting data into an interdependency-interaction model of a complex. We applied this approach to a data set generated from components of the Saccharomyces cerevisiae Rpd3 histone deacetylase complexes, which consists of two distinct small and large complexes that are held together by a module consisting of Rpd3, Sin3 and Ume1. The resulting representation reveals new protein-protein interactions and new submodule relationships, providing novel information for mapping the functional organization of a complex. PMID- 19806190 TI - Resilience of coral-associated bacterial communities exposed to fish farm effluent. AB - BACKGROUND: The coral holobiont includes the coral animal, algal symbionts, and associated microbial community. These microbes help maintain the holobiont homeostasis; thus, sustaining robust mutualistic microbial communities is a fundamental part of long-term coral reef survival. Coastal pollution is one major threat to reefs, and intensive fish farming is a rapidly growing source of this pollution. METHODOLOGY & PRINCIPAL FINDINGS: We investigated the susceptibility and resilience of the bacterial communities associated with a common reef building coral, Porites cylindrica, to coastal pollution by performing a clonally replicated transplantation experiment in Bolinao, Philippines adjacent to intensive fish farming. Ten fragments from each of four colonies (total of 40 fragments) were followed for 22 days across five sites: a well-flushed reference site (the original fragment source); two sites with low exposure to milkfish (Chanos chanos) aquaculture effluent; and two sites with high exposure. Elevated levels of dissolved organic carbon (DOC), chlorophyll a, total heterotrophic and autotrophic bacteria abundance, virus like particle (VLP) abundances, and culturable Vibrio abundance characterized the high effluent sites. Based on 16S rRNA clone libraries and denaturing gradient gel electrophoresis (DGGE) analysis, we observed rapid, dramatic changes in the coral-associated bacterial communities within five days of high effluent exposure. The community composition on fragments at these high effluent sites shifted towards known human and coral pathogens (i.e. Arcobacter, Fusobacterium, and Desulfovibrio) without the host corals showing signs of disease. The communities shifted back towards their original composition by day 22 without reduction in effluent levels. SIGNIFICANCE: This study reveals fish farms as a likely source of pathogens with the potential to proliferate on corals and an unexpected short-term resilience of coral-associated bacterial communities to eutrophication pressure. These data highlight a need for improved aquaculture practices that can achieve both sustainable industry goals and long-term coral reef survival. PMID- 19806191 TI - Inherited glutathione reductase deficiency and Plasmodium falciparum malaria--a case study. AB - In Plasmodium falciparum-infected red blood cells (RBCs), the flavoenzyme glutathione reductase (GR) regenerates reduced glutathione, which is essential for antioxidant defense. GR utilizes NADPH produced in the pentose phosphate shunt by glucose-6-phosphate dehydrogenase (G6PD). Thus, conditions affecting host G6PD or GR induce increased sensitivity to oxidants. Hereditary G6PD deficiency is frequent in malaria endemic areas and provides protection against severe malaria. Furthermore, GR deficiency resulting from insufficient saturation of the enzyme with its prosthetic group FAD is common. Based on these naturally occurring phenomena, GR of malaria parasites and their host cells represent attractive antimalarial drug targets. Recently we were given the opportunity to examine invasion, growth, and drug sensitivity of three P. falciparum strains (3D7, K1, and Palo Alto) in the RBCs from three homozygous individuals with total GR deficiency resulting from mutations in the apoprotein. Invasion or growth in the GR-deficient RBCs was not impaired for any of the parasite strains tested. Drug sensitivity to chloroquine, artemisinin, and methylene blue was comparable to parasites grown in GR-sufficient RBCs and sensitivity towards paraquat and sodium nitroprusside was only slightly enhanced. In contrast, membrane deposition of hemichromes as well as the opsonizing complement C3b fragments and phagocytosis were strongly increased in ring-infected RBCs of the GR-deficient individuals compared to ring-infected normal RBCs. Also, in one of the individuals, membrane-bound autologous IgGs were significantly enhanced. Thus, based on our in vitro data, GR deficiency and drug-induced GR inhibition may protect from malaria by inducing enhanced ring stage phagocytosis rather than by impairing parasite growth directly. PMID- 19806192 TI - Innate immune responses of pulmonary epithelial cells to Burkholderia pseudomallei infection. AB - BACKGROUND: Burkholderia pseudomallei, a facultative intracellular pathogen, causes systemic infection in humans with high mortality especially when infection occurs through an infectious aerosol. Previous studies indicated that the epithelial cells in the lung are an active participant in host immunity. In this study, we aimed to investigate the innate immune responses of lung epithelial cells against B. pseudomallei. METHODOLOGY AND PRINCIPAL FINDINGS: Using a murine lung epithelial cell line, primary lung epithelial cells and an inhalational murine infection model, we characterized the types of innate immunity proteins and peptides produced upon B. pseudomallei infection. Among a wide panel of immune components studied, increased levels of major pro-inflammatory cytokines IL-6 and TNFalpha, chemokine MCP-1, and up-regulation of secretory leukocyte protease inhibitor (SLPI) and chemokine (C-C motif) ligand 20 (CCL20) were observed. Inhibition assays using specific inhibitors suggested that NF-kappaB and p38 MAPK pathways were responsible for these B. pseudomallei-induced antimicrobial peptides. CONCLUSIONS: Our findings indicate that the respiratory epithelial cells, which form the majority of the cells lining the epithelial tract and the lung, have important roles in the innate immune response against B. pseudomallei infection. PMID- 19806193 TI - Characterization of multiple ion channels in cultured human cardiac fibroblasts. AB - BACKGROUND: Although fibroblast-to-myocyte electrical coupling is experimentally suggested, electrophysiology of cardiac fibroblasts is not as well established as contractile cardiac myocytes. The present study was therefore designed to characterize ion channels in cultured human cardiac fibroblasts. METHODS AND FINDINGS: A whole-cell patch voltage clamp technique and RT-PCR were employed to determine ion channels expression and their molecular identities. We found that multiple ion channels were heterogeneously expressed in human cardiac fibroblasts. These include a big conductance Ca(2+)-activated K(+) current (BK(Ca)) in most (88%) human cardiac fibroblasts, a delayed rectifier K(+) current (IK(DR)) and a transient outward K(+) current (I(to)) in a small population (15 and 14%, respectively) of cells, an inwardly-rectifying K(+) current (I(Kir)) in 24% of cells, and a chloride current (I(Cl)) in 7% of cells under isotonic conditions. In addition, two types of voltage-gated Na(+) currents (I(Na)) with distinct properties were present in most (61%) human cardiac fibroblasts. One was a slowly inactivated current with a persistent component, sensitive to tetrodotoxin (TTX) inhibition (I(Na.TTX), IC(50) = 7.8 nM), the other was a rapidly inactivated current, relatively resistant to TTX (I(Na.TTXR), IC(50) = 1.8 microM). RT-PCR revealed the molecular identities (mRNAs) of these ion channels in human cardiac fibroblasts, including KCa.1.1 (responsible for BK(Ca)), Kv1.5, Kv1.6 (responsible for IK(DR)), Kv4.2, Kv4.3 (responsible for I(to)), Kir2.1, Kir2.3 (for I(Kir)), Clnc3 (for I(Cl)), Na(V)1.2, Na(V)1.3, Na(V)1.6, Na(V)1.7 (for I(Na.TTX)), and Na(V)1.5 (for I(Na.TTXR)). CONCLUSIONS: These results provide the first information that multiple ion channels are present in cultured human cardiac fibroblasts, and suggest the potential contribution of these ion channels to fibroblast-myocytes electrical coupling. PMID- 19806194 TI - Two alleles of NF-kappaB in the sea anemone Nematostella vectensis are widely dispersed in nature and encode proteins with distinct activities. AB - BACKGROUND: NF-kappaB is an evolutionarily conserved transcription factor that controls the expression of genes involved in many key organismal processes, including innate immunity, development, and stress responses. NF-kappaB proteins contain a highly conserved DNA-binding/dimerization domain called the Rel homology domain. METHODS/PRINCIPAL FINDINGS: We characterized two NF-kappaB alleles in the sea anemone Nematostella vectensis that differ at nineteen single nucleotide polymorphisms (SNPs). Ten of these SNPs result in amino acid substitutions, including six within the Rel homology domain. Both alleles are found in natural populations of Nematostella. The relative abundance of the two NF-kappaB alleles differs between populations, and departures from Hardy-Weinberg equilibrium within populations indicate that the locus may be under selection. The proteins encoded by the two Nv-NF-kappaB alleles have different molecular properties, in part due to a Cys/Ser polymorphism at residue 67, which resides within the DNA recognition loop. In nearly all previously characterized NF-kappaB proteins, the analogous residue is fixed for Cys, and conversion of human RHD proteins from Cys to Ser at this site has been shown to increase DNA-binding ability and increase resistance to inhibition by thiol-reactive compounds. However, the naturally-occurring Nematostella variant with Cys at position 67 binds DNA with a higher affinity than the Ser variant. On the other hand, the Ser variant activates transcription in reporter gene assays more effectively, and it is more resistant to inhibition by a thiol-reactive compound. Reciprocal Cys< >Ser mutations at residue 67 of the native Nv-NF-kappaB proteins affect DNA binding as in human NF-kappaB proteins, e.g., a Cys->Ser mutation increases DNA binding of the native Cys variant. CONCLUSIONS/SIGNIFICANCE: These results are the first demonstration of a naturally occurring and functionally significant polymorphism in NF-kappaB in any species. The functional differences between these alleles and their uneven distribution in the wild suggest that different genotypes could be favored in different environments, perhaps environments that vary in their levels of peroxides or thiol-reactive compounds. PMID- 19806195 TI - Characterization of a natural mutator variant of human DNA polymerase lambda which promotes chromosomal instability by compromising NHEJ. AB - BACKGROUND: DNA polymerase lambda (Pollambda) is a DNA repair polymerase, which likely plays a role in base excision repair (BER) and in non-homologous end joining (NHEJ) of DNA double-strand breaks (DSB). PRINCIPAL FINDINGS: Here, we described a novel natural allelic variant of human Pollambda (hPollambda) characterized by a single nucleotide polymorphism (SNP), C/T variation in the first base of codon 438, resulting in the amino acid change Arg to Trp. In vitro enzyme activity assays of the purified W438 Pollambda variant revealed that it retained both DNA polymerization and deoxyribose phosphate (dRP) lyase activities, but had reduced base substitution fidelity. Ectopic expression of the W438 hPollambda variant in mammalian cells increases mutation frequency, affects the DSB repair NHEJ pathway, and generates chromosome aberrations. All these phenotypes are dependent upon the catalytic activity of the W438 hPollambda. CONCLUSIONS: The expression of a cancer-related natural variant of one specialized DNA polymerase can be associated to generic instability at the cromosomal level, probably due a defective NHEJ. These results establish that chromosomal aberrations can result from mutations in specialized DNA repair polymerases. PMID- 19806196 TI - A conditioning lesion provides selective protection in a rat model of Amyotrophic Lateral Sclerosis. AB - BACKGROUND: Amyotrophic Lateral Sclerosis (ALS) is neurodegenerative disease characterized by muscle weakness and atrophy due to progressive motoneuron loss. The death of motoneuron is preceded by the failure of neuromuscular junctions (NMJs) and axonal retraction. Thus, to develop an effective ALS therapy you must simultaneously preserve motoneuron somas, motor axons and NMJs. A conditioning lesion has the potential to accomplish this since it has been shown to enhance neuronal survival and recovery from trauma in a variety of contexts. METHODOLOGY/PRINCIPAL FINDINGS: To test the effects of a conditioning lesion in a model of familial ALS we administered a tibial nerve crush injury to presymptomatic fALS(G93A) rats. We examined its effects on motor function, motoneuron somas, motor axons, and NMJs. Our experiments revealed a novel paradigm for the conditioning lesion effect. Specifically we found that the motor functional decline in fALS(G93A) rats that received a conditioning lesion was delayed and less severe. These improvements in motor function corresponded to greater motoneuron survival, reduced motor axonopathy, and enhanced NMJ maintenance at disease end-stage. Furthermore, the increased NMJ maintenance was selective for muscle compartments innervated by the most resilient (slow) motoneuron subtypes, but was absent in muscle compartments innervated by the most vulnerable (fast fatigable) motoneuron subtypes. CONCLUSIONS/SIGNIFICANCE: These findings support the development of strategies aimed at mimicking the conditioning lesion effect to treat ALS as well as underlined the importance of considering the heterogeneity of motoneuron sub-types when evaluating prospective ALS therapeutics. PMID- 19806197 TI - Evidence for the emergence of new rice types of interspecific hybrid origin in West African farmers' fields. AB - In West Africa two rice species (Oryza glaberrima Steud. and Oryza sativa L.) co exist. Although originally it was thought that interspecific hybridization is impossible without biotechnological methods, progenies of hybridization appear to occur in farmer fields. AFLP analysis was used to assess genetic diversity in West Africa (including the countries The Gambia, Senegal, Guinea Bissau, Guinea Conakry, Sierra Leone, Ghana and Togo) using 315 rice samples morphologically classified prior to analysis. We show evidence for farmer interspecific hybrids of African and Asian rice, resulting in a group of novel genotypes, and identify possible mechanisms for in-field hybridization. Spontaneous back-crossing events play a crucial role, resulting in different groups of genetic diversity in different regions developed by natural and cultural selection, often under adverse conditions. These new groups of genotypes may have potential relevance for exploitation by plant breeders. Future advances in crop development could be achieved through co-operation between scientists and marginalized farmer groups in order to address challenges of rapid adaptation in a world of increasing socio political and climatic uncertainty. PMID- 19806198 TI - Muscle-specific adaptations, impaired oxidative capacity and maintenance of contractile function characterize diet-induced obese mouse skeletal muscle. AB - BACKGROUND: The effects of diet-induced obesity on skeletal muscle function are largely unknown, particularly as it relates to changes in oxidative metabolism and morphology. PRINCIPAL FINDINGS: Compared to control fed mice, mice fed a high fat diet (HFD; 60% kcal: fat) for 8 weeks displayed increased body mass and insulin resistance without overt fasting hyperglycemia (i.e. pre-diabetic). Histological analysis revealed a greater oxidative potential in the HFD gastrocnemius/plantaris (increased IIA, reduced IIB fiber-type percentages) and soleus (increased I, IIA cross-sectional areas) muscles, but no change in fiber type percentages in tibialis anterior muscles compared to controls. Intramyocellular lipid levels were significantly increased relative to control in HFD gastrocnemius/plantaris, but were similar to control values in the HFD soleus. Using a novel, single muscle fiber approach, impairments in complete palmitate and glucose oxidation (72.8+/-6.6% and 61.8+/-9.1% of control, respectively; p<0.05) with HFD were detected. These reductions were consistent with measures made using intact extensor digitorum longus and soleus muscles. Compared to controls, no difference in succinate dehydrogenase or citrate synthase enzyme activities were observed between groups in any muscle studied, however, short-chain fatty acyl CoA dehydrogenase (SCHAD) activity was elevated in the HFD soleus, but not tibialis anterior muscles. Despite these morphological and metabolic alterations, no significant difference in peak tetanic force or low frequency fatigue rates were observed between groups. CONCLUSIONS: These findings indicate that HFD induces early adaptive responses that occur in a muscle specific pattern, but are insufficient to prevent impairments in oxidative metabolism with continued high-fat feeding. Moreover, the morphological and metabolic changes which occur with 8 weeks of HFD do not significantly impact muscle contractile properties. PMID- 19806199 TI - Insights into the molecular basis of L-form formation and survival in Escherichia coli. AB - L-forms have been shown to occur among many species of bacteria and are suspected to be involved in persistent infections. Since their discovery in 1935, numerous studies characterizing L-form morphology, growth, and pathogenic potential have been conducted. However, the molecular mechanisms underlying the formation and survival of L-forms remain unknown. Using unstable L-form colonies of Escherichia coli as a model, we performed genome-wide transcriptome analysis and screened a deletion mutant library to study the molecular mechanisms involved in formation and survival of L-forms. Microarray analysis of L-form versus classical colonies revealed many up-regulated genes of unknown function as well as multiple over expressed stress pathways shared in common with persister cells and biofilms. Mutant screens identified three groups of mutants which displayed varying degrees of defects in L-form colony formation. Group 1 mutants, which showed the strongest defect in L-form colony formation, belonged to pathways involved in cell envelope stress, DNA repair, iron homeostasis, outer membrane biogenesis, and drug efflux/ABC transporters. Four (Group 1) mutants, rcsB, a positive response regulator of colanic acid capsule synthesis, ruvA, a recombinational junction binding protein, fur, a ferric uptake regulator and smpA a small membrane lipoprotein were selected for complementation. Complementation of the mutants using a high-copy overexpression vector failed, while utilization of a low-copy inducible vector successfully restored L-form formation. This work represents the first systematic genetic evaluation of genes and pathways involved in the formation and survival of unstable L-form bacteria. Our findings provide new insights into the molecular mechanisms underlying L-form formation and survival and have implications for understanding the emergence of antibiotic resistance, bacterial persistence and latent infections and designing novel drugs and vaccines. PMID- 19806200 TI - Activin/Nodal inhibition alone accelerates highly efficient neural conversion from human embryonic stem cells and imposes a caudal positional identity. AB - BACKGROUND: Neural conversion from human embryonic stem cells (hESCs) has been demonstrated in a variety of systems including chemically defined suspension culture, not requiring extrinsic signals, as well as in an adherent culture method that involves dual SMAD inhibition using Noggin and SB431542 (an inhibitor of activin/nodal signaling). Previous studies have also determined a role for activin/nodal signaling in development of the neural plate and anterior fate specification. We therefore sought to investigate the independent influence of SB431542 both on neural commitment of hESCs and positional identity of derived neural progenitors in chemically defined substrate-free conditions. METHODOLOGY/PRINCIPAL FINDINGS: We show that in non-adherent culture conditions, treatment with SB431542 alone for 8 days is sufficient for highly efficient and accelerated neural conversion from hESCs with negligible mesendodermal, epidermal or trophectodermal contamination. In addition the resulting neural progenitor population has a predominantly caudal identity compared to the more anterior positional fate of non-SB431542 treated cultures. Finally we demonstrate that resulting neurons are electro-physiologically competent. CONCLUSIONS: This study provides a platform for the efficient generation of caudal neural progenitors under defined conditions for experimental study. PMID- 19806201 TI - Basal cancer cell survival involves JNK2 suppression of a novel JNK1/c-Jun/Bcl-3 apoptotic network. AB - BACKGROUND: The regulation of apoptosis under basal (non-stress) conditions is crucial for normal mammalian development and also for normal cellular turnover in different tissues throughout life. Deficient regulation of basal apoptosis, or its perturbation, can result in impaired development and/or disease states including cancer. In contrast to stress-induced apoptosis the regulation of apoptosis under basal conditions is poorly understood. To address this issue we have compared basal- and stress-induced apoptosis in human epithelial cells of normal and cancerous origins. For this purpose we focussed our study on the opposing pro-apoptotic JNK/anti-apoptotic NFkappaB pathways. METHODOLOGY/PRINCIPAL FINDINGS: Combinatorial RNAi plus gene knockout were employed to access and map basal regulatory pathways of apoptosis. Follow-on, in depth analyses included exogenous expression of phosphorylation mutants and chromatin immunoprecipitation. We demonstrate that basal apoptosis is constitutively suppressed by JNK2 in a range of human cancer cell lines. This effect was not observed in non-cancer cells. Silencing JNK2 by RNAi resulted in JNK1-dependent apoptosis of cancer cells via up-regulation of the AP-1 factor c Jun. Unexpectedly we discovered that JNK1 and c-Jun promote basal apoptosis in the absence of "activating phosphorylations" typically induced by stress. Hypo phosphorylated c-Jun accumulated to high levels following JNK2 silencing, auto regulated its own expression and suppressed expression of Bcl-3, an unusual IkappaB protein and regulator of NFkappaB. Basal apoptosis was mediated by components of the TNFalpha response pathway but was mechanistically distinct from TNFalpha-induced apoptosis. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that mechanistically distinct pathways operate to regulate apoptosis in mammalian cells under basal (physiological) versus stress-induced conditions. We also describe a novel apoptotic network which governs the basal survival of cancer cells. Such information is crucial for understanding normal cellular turnover during mammalian development and subsequently throughout life. This information also opens new avenues for therapeutic intervention in human proliferative disease states including cancer. PMID- 19806203 TI - The production of antibody by invading B cells is required for the clearance of rabies virus from the central nervous system. AB - BACKGROUND: The pathogenesis of rabies is associated with the inability to deliver immune effectors across the blood-brain barrier and to clear virulent rabies virus from CNS tissues. However, the mechanisms that facilitate immune effector entry into CNS tissues are induced by infection with attenuated rabies virus. METHODOLOGY/PRINCIPAL FINDINGS: Infection of normal mice with attenuated rabies virus but not immunization with killed virus can promote the clearance of pathogenic rabies virus from the CNS. T cell activity in B cell-deficient mice can control the replication of attenuated virus in the CNS, but viral mRNA persists. Low levels of passively administered rabies virus-neutralizing antibody reach infected cells in the cerebellum of B cell-deficient mice but are not sufficient to mediate virus clearance. Production of rabies virus-specific antibody by B cells invading CNS tissues is required for this process, and a substantial proportion of the B cells that accumulate in the CNS of mice infected with attenuated rabies virus produce virus-specific antibodies. CONCLUSIONS/SIGNIFICANCE: The mechanisms required for immune effectors to enter rabies virus-infected tissues are induced by infection with attenuated rabies virus but not by infection with pathogenic rabies viruses or immunization with killed virus. T cell activities can inhibit rabies virus replication, but the production of rabies virus-specific antibodies by infiltrating B cells, as opposed to the leakage of circulating antibody across the BBB, is critical to elimination of the virus. These findings suggest that a pathogenic rabies virus infection may be treatable after the virus has reached the CNS tissues, providing that the appropriate immune effectors can be targeted to the infected tissues. PMID- 19806204 TI - Brugia malayi gene expression in response to the targeting of the Wolbachia endosymbiont by tetracycline treatment. AB - BACKGROUND: Brugia malayi, like most human filarial parasite species, harbors an endosymbiotic bacterium of the genus Wolbachia. Elimination of the endosymbiont leads to sterilization of the adult female. Previous biochemical and genetic studies have established that communication with its endobacterium is essential for survival of the worm. METHODOLOGY/PRINCIPAL FINDINGS: We used electron microscopy to examine the effects of antibiotic treatment on Wolbachia cell structure. We have also used microarray and quantitative RT-PCR analyses to examine the regulation of the B. malayi transcripts altered in response to the anti-Wolbachia treatment. Microscopy of worms taken from animals treated with tetracycline for 14 and 21 days (14 d and 21 d) demonstrated substantial morphologic effects on the Wolbachia endobacterium by 14 d and complete degeneration of the endobacterial structures by 21 d. We observed upregulation of transcripts primarily encoding proteins involved in amino acid synthesis and protein translation, and downregulation of transcripts involved in cuticle biosynthesis after both 7 d and 14 d of treatment. In worms exposed to tetracycline in culture, substantial effects on endobacteria morphology were evident by day 3, and extensive death of the endobacteria was observed by day 5. In a detailed examination of the expression kinetics of selected signaling genes carried out on such cultured worms, a bimodal pattern of regulation was observed. The selected genes were upregulated during the early phase of antibiotic treatment and quickly downregulated in the following days. These same genes were upregulated once more at 6 days post-treatment. CONCLUSIONS/SIGNIFICANCE: Upregulation of protein translation and amino acid synthesis may indicate a generalized stress response induced in B. malayi due to a shortage of essential nutrients/factors that are otherwise supplied by Wolbachia. Downregulation of transcripts involved in cuticle biosynthesis perhaps reflects a disruption in the normal embryogenic program. This is confirmed by the expression pattern of transcripts that may be representative of the worms' response to Wolbachia in different tissues; the early peak potentially reflects the effect of bacteria death on the embryogenic program while the second peak may be a manifestation of the adult worm response to the affected bacteria within the hypodermis. PMID- 19806205 TI - Widespread distribution of a newly found point mutation in voltage-gated sodium channel in pyrethroid-resistant Aedes aegypti populations in Vietnam. AB - BACKGROUND: Resistance of Aedes aegypti to photostable pyrethroid insecticides is a major problem for disease-vector control programs. Pyrethroids target the voltage-gated sodium channel on the insects' neurons. Single amino acid substitutions in this channel associated with pyrethroid resistance are one of the main factors that cause knockdown resistance in insects. Although kdr has been observed in several mosquito species, point mutations in the para gene have not been fully characterized in Ae. aegypti populations in Vietnam. The aim of this study was to determine the types and frequencies of mutations in the para gene in Ae. aegypti collected from used tires in Vietnam. METHODS AND FINDINGS: Several point mutations were examined that cause insensitivity of the voltage gated sodium channel in the insect nervous system due to the replacement of the amino acids L1014F, the most commonly found point mutation in several mosquitoes; I1011M (or V) and V1016G (or I), which have been reported to be associated to knockdown resistance in Ae. aegypti located in segment 6, domain II; and a recently found amino acid replacement in F1269 in Ae. aegypti, located in segment 6, domain III. Among 756 larvae from 70 locations, no I1011M or I1011V nor L1014F mutations were found, and only two heterozygous V1016G mosquitoes were detected. However, F1269C mutations on domain III were distributed widely and with high frequency in 269 individuals among 757 larvae (53 collection sites among 70 locations surveyed). F1269C frequencies were low in the middle to north part of Vietnam but were high in the areas neighboring big cities and in the south of Vietnam, with the exception of the southern mountainous areas located at an elevation of 500-1000 m. CONCLUSIONS: The overall percentage of homozygous F1269C seems to remain low (7.4%) in the present situation. However, extensive and uncontrolled frequent use of photostable pyrethroids might be a strong selection pressure for this mutation to cause serious problems in the control of dengue fever in Vietnam. PMID- 19806206 TI - The repertoire of heterotrimeric G proteins and RGS proteins in Ciona intestinalis. AB - BACKGROUND: Heterotrimeric G proteins and regulators of G protein signaling (RGS) proteins are key downstream interacting partners in the G protein coupled receptor (GPCR) signaling pathway. The highly versatile GPCR transmembrane signaling system is a consequence of the coupling of a diverse set of receptors to downstream partners that include multiple subforms of G proteins and regulatory proteins including RGS proteins, among others. While the GPCR repertoire of Ciona intestinalis, representing the basal chordate is known, the repertoire of the heterotrimeric G proteins and RGS proteins is unknown. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we performed an in-silico genome-wide search of C. intestinalis for its complement of G proteins and RGS proteins. The identification of several one-to-one orthologs of human G proteins at the levels of families, subfamilies and types and of homologs of the human RGS proteins suggests an evolutionarily conserved structure function relationship of the GPCR signaling mechanism in the chordates. CONCLUSIONS: The C. intestinalis genome encodes a highly conserved, albeit, limited repertoire of the heterotrimeric G protein complexes with the size of subunit types comparable with that in lower eukaryotes. PMID- 19806207 TI - An efficient targeted drug delivery through apotransferrin loaded nanoparticles. AB - BACKGROUND: Cancerous state is a highly stimulated environment of metabolically active cells. The cells under these conditions over express selective receptors for assimilation of factors essential for growth and transformation. Such receptors would serve as potential targets for the specific ligand mediated transport of pharmaceutically active molecules. The present study demonstrates the specificity and efficacy of protein nanoparticle of apotransferrin for targeted delivery of doxorubicin. METHODOLOGY/PRINCIPAL FINDINGS: Apotransferrin nanoparticles were developed by sol-oil chemistry. A comparative analysis of efficiency of drug delivery in conjugated and non-conjugated forms of doxorubicin to apotransferrin nanoparticle is presented. The spherical shaped apotransferrin nanoparticles (nano) have diameters of 25-50 etam, which increase to 60-80 etam upon direct loading of drug (direct-nano), and showed further increase in dimension (75-95 etam) in conjugated nanoparticles (conj-nano). The competitive experiments with the transferrin receptor specific antibody showed the entry of both conj-nano and direct-nano into the cells through transferrin receptor mediated endocytosis. Results of various studies conducted clearly establish the superiority of the direct-nano over conj-nano viz. (a) localization studies showed complete release of drug very early, even as early as 30 min after treatment, with the drug localizing in the target organelle (nucleus) (b) pharmacokinetic studies showed enhanced drug concentrations, in circulation with sustainable half-life (c) the studies also demonstrated efficient drug delivery, and an enhanced inhibition of proliferation in cancer cells. Tissue distribution analysis showed intravenous administration of direct nano lead to higher drug localization in liver, and blood as compared to relatively lesser localization in heart, kidney and spleen. Experiments using rat cancer model confirmed the efficacy of the formulation in regression of hepatocellular carcinoma with negligible toxicity to kidney and liver. CONCLUSIONS: The present study thus demonstrates that the direct-nano is highly efficacious in delivery of drug in a target specific manner with lower toxicity to heart, liver and kidney. PMID- 19806208 TI - Vasoregression linked to neuronal damage in the rat with defect of polycystin-2. AB - BACKGROUND: Neuronal damage is correlated with vascular dysfunction in the diseased retina, but the underlying mechanisms remain controversial because of the lack of suitable models in which vasoregression related to neuronal damage initiates in the mature retinal vasculature. The aim of this study was to assess the temporal link between neuronal damage and vascular patency in a transgenic rat (TGR) with overexpression of a mutant cilia gene polycystin-2. METHODS: Vasoregression, neuroglial changes and expression of neurotrophic factors were assessed in TGR and control rats in a time course. Determination of neuronal changes was performed by quantitative morphometry of paraffin-embedded vertical sections. Vascular cell composition and patency were assessed by quantitative retinal morphometry of digest preparations. Glial activation was assessed by western blot and immunofluorescence. Expression of neurotrophic factors was detected by quantitative PCR. FINDINGS: At one month, number and thickness of the outer nuclear cell layers (ONL) in TGR rats were reduced by 31% (p<0.001) and 17% (p<0.05), respectively, compared to age-matched control rats. Furthermore, the reduction progressed from 1 to 7 months in TGR rats. Apoptosis was selectively detected in the photoreceptor in the ONL, starting after one month. Nevertheless, TGR and control rats showed normal responses in electroretinogram at one month. From the second month onwards, TGR retinas had significantly increased acellular capillaries (p<0.001), and a reduction of endothelial cells (p<0.01) and pericytes (p<0.01). Upregulation of GFAP was first detected in TGR retinas after 1 month in glial cells, in parallel with an increase of FGF2 (fourfold) and CNTF (60 %), followed by upregulation of NGF (40 %) at 3 months. INTERPRETATION: Our data suggest that TGR is an appropriate animal model for vasoregression related to neuronal damage. Similarities to experimental diabetic retinopathy render this model suitable to understand general mechanisms of maturity-onset vasoregression. PMID- 19806209 TI - High expression levels of total IGF-1R and sensitivity of NSCLC cells in vitro to an anti-IGF-1R antibody (R1507). AB - BACKGROUND: The IGF receptor type 1 (IGF-1R) pathway is frequently deregulated in human tumors and has become a target of interest for anti-cancer therapy. METHODOLOGY/PRINCIPAL FINDINGS: We used a panel of 22 non-small cell lung cancer (NSCLC) cell lines to investigate predictive biomarkers of response to R1507, a fully-humanized anti-IGF-1R monoclonal antibody (Ab; Roche). 5 lines were moderately sensitive (25-50% growth inhibition) to R1507 alone. While levels of phospho-IGF-1R did not correlate with drug sensitivity, 4 out of 5 sensitive lines displayed high levels of total IGF-1R versus 1 out of 17 resistant lines (p = 0.003, Fisher's Exact). Sensitive lines also harbored higher copy numbers of IGF-1R as assessed by independent SNP array analysis. Addition of erlotinib or paclitaxel to R1507 led to further growth inhibition in sensitive but not resistant lines. In one EGFR mutant lung adenocarcinoma cell line (11-18), R1507 and erlotinib co-treatment induced apoptosis, whereas treatment with either drug alone induced only cell cycle arrest. Apoptosis was mediated, in part, by the survival-related AKT pathway. Additionally, immunohistochemical (IHC) staining of total IGF-1R with an anti-total IGF-1R Ab (G11;Ventana) was performed on tissue microarrays (TMAs) containing 270 independent NSCLC tumor samples. Staining intensity was scored on a scale of 0 to 3+. 39.3% of tumors showed medium to high IGF-1R IHC staining (scores of 2+ or 3+, respectively), while 16.7% had scores of 3+. CONCLUSIONS/SIGNIFICANCE: In NSCLC cell lines, high levels of total IGF-1R are associated with moderate sensitivity to R1507. These results suggest a possible enrichment strategy for clinical trials with anti-IGF-1R therapy. PMID- 19806210 TI - Costs and effects of abdominal versus laparoscopic hysterectomy: systematic review of controlled trials. AB - OBJECTIVE: Comparative evaluation of costs and effects of laparoscopic hysterectomy (LH) and abdominal hysterectomy (AH). DATA SOURCES: Controlled trials from Cochrane Central register of controlled trials, Medline, Embase and prospective trial registers. SELECTION OF STUDIES: Twelve (randomized) controlled studies including the search terms costs, laparoscopy, laparotomy and hysterectomy were identified. METHODS: The type of cost analysis, perspective of cost analyses and separate cost components were assessed. The direct and indirect costs were extracted from the original studies. For the cost estimation, hospital stay and procedure costs were selected as most important cost drivers. As main outcome the major complication rate was taken. FINDINGS: Analysis was performed on 2226 patients, of which 1013 (45.5%) in the LH group and 1213 (54.5%) in the AH group. Five studies scored > or =10 points (out of 19) for methodological quality. The reported total direct costs in the LH group ($63,997) were 6.1% higher than the AH group ($60,114). The reported total indirect costs of the LH group ($1,609) were half of the total indirect in the AH group ($3,139). The estimated mean major complication rate in the LH group (14.3%) was lower than in the AH group (15.9%). The estimated total costs in the LH group were $3,884 versus $3,312 in the AH group. The incremental costs for reducing one patient with major complication(s) in the LH group compared to the AH group was $35,750. CONCLUSIONS: The shorter hospital stay in the LH group compensates for the increased procedure costs, with less morbidity. LH points in the direction of cost effectiveness, however further research is warranted with a broader costs perspective including long term effects as societal benefit, quality of life and survival. PMID- 19806211 TI - Phagocytosis of Streptococcus pyogenes by all-trans retinoic acid-differentiated HL-60 cells: roles of azurophilic granules and NADPH oxidase. AB - BACKGROUND: New experimental approaches to the study of the neutrophil phagosome and bacterial killing prompted a reassessment of the usefulness of all-trans retinoic acid (ATRA)-differentiated HL-60 cells as a neutrophil model. HL-60 cells are special in that they possess azurophilic granules while lacking the specific granules with their associated oxidase components. The resulting inability to mount an effective intracellular respiratory burst makes these cells more dependent on other mechanisms when killing internalized bacteria. METHODOLOGY/PRINCIPAL FINDINGS: In this work phagocytosis and phagosome-related responses of ATRA-differentiated HL-60 cells were compared to those earlier described in human neutrophils. We show that intracellular survival of wild-type S. pyogenes bacteria in HL-60 cells is accompanied by inhibition of azurophilic granule-phagosome fusion. A mutant S. pyogenes bacterium, deficient in M-protein expression, is, on the other hand, rapidly killed in phagosomes that avidly fuse with azurophilic granules. CONCLUSIONS/SIGNIFICANCE: The current data extend our previous findings by showing that a system lacking in oxidase involvement also indicates a link between inhibition of azurophilic granule fusion and the intraphagosomal fate of S. pyogenes bacteria. We propose that differentiated HL 60 cells can be a useful tool to study certain aspects of neutrophil phagosome maturation, such as azurophilic granule fusion. PMID- 19806212 TI - Phylogenetic analysis of seven WRKY genes across the palm subtribe Attaleinae (Arecaceae) [corrected] identifies Syagrus as sister group of the coconut. AB - BACKGROUND: The Cocoseae is one of 13 tribes of Arecaceae subfam. Arecoideae, and contains a number of palms with significant economic importance, including the monotypic and pantropical Cocos nucifera L., the coconut, the origins of which have been one of the "abominable mysteries" of palm systematics for decades. Previous studies with predominantly plastid genes weakly supported American ancestry for the coconut but ambiguous sister relationships. In this paper, we use multiple single copy nuclear loci to address the phylogeny of the Cocoseae subtribe Attaleinae, and resolve the closest extant relative of the coconut. METHODOLOGY/PRINCIPAL FINDINGS: We present the results of combined analysis of DNA sequences of seven WRKY transcription factor loci across 72 samples of Arecaceae tribe Cocoseae subtribe Attaleinae, representing all genera classified within the subtribe, and three outgroup taxa with maximum parsimony, maximum likelihood, and Bayesian approaches, producing highly congruent and well-resolved trees that robustly identify the genus Syagrus as sister to Cocos and resolve novel and well-supported relationships among the other genera of the Attaleinae. We also address incongruence among the gene trees with gene tree reconciliation analysis, and assign estimated ages to the nodes of our tree. CONCLUSIONS/SIGNIFICANCE: This study represents the as yet most extensive phylogenetic analyses of Cocoseae subtribe Attaleinae. We present a well-resolved and supported phylogeny of the subtribe that robustly indicates a sister relationship between Cocos and Syagrus. This is not only of biogeographic interest, but will also open fruitful avenues of inquiry regarding evolution of functional genes useful for crop improvement. Establishment of two major clades of American Attaleinae occurred in the Oligocene (ca. 37 MYBP) in Eastern Brazil. The divergence of Cocos from Syagrus is estimated at 35 MYBP. The biogeographic and morphological congruence that we see for clades resolved in the Attaleinae suggests that WRKY loci are informative markers for investigating the phylogenetic relationships of the palm family. PMID- 19806213 TI - Dynamic locomotor capabilities revealed by early dinosaur trackmakers from southern Africa. AB - BACKGROUND: A new investigation of the sedimentology and ichnology of the Early Jurassic Moyeni tracksite in Lesotho, southern Africa has yielded new insights into the behavior and locomotor dynamics of early dinosaurs. METHODOLOGY/PRINCIPAL FINDINGS: The tracksite is an ancient point bar preserving a heterogeneous substrate of varied consistency and inclination that includes a ripple-marked riverbed, a bar slope, and a stable algal-matted bar top surface. Several basal ornithischian dinosaurs and a single theropod dinosaur crossed its surface within days or perhaps weeks of one another, but responded to substrate heterogeneity differently. Whereas the theropod trackmaker accommodated sloping and slippery surfaces by gripping the substrate with its pedal claws, the basal ornithischian trackmakers adjusted to the terrain by changing between quadrupedal and bipedal stance, wide and narrow gauge limb support (abduction range = 31 degrees ), and plantigrade and digitigrade foot posture. CONCLUSIONS/SIGNIFICANCE: The locomotor adjustments coincide with changes in substrate consistency along the trackway and appear to reflect 'real time' responses to a complex terrain. It is proposed that these responses foreshadow important locomotor transformations characterizing the later evolution of the two main dinosaur lineages. Ornithischians, which shifted from bipedal to quadrupedal posture at least three times in their evolutionary history, are shown to have been capable of adopting both postures early in their evolutionary history. The substrate-gripping behavior demonstrated by the early theropod, in turn, is consistent with the hypothesized function of pedal claws in bird ancestors. PMID- 19806214 TI - Adenotonsillectomy and neurocognitive deficits in children with Sleep Disordered Breathing. AB - BACKGROUND: Sleep Disordered Breathing (SDB) is a common childhood disorder that encompasses a range of sleep-related upper airway obstruction. Children with SDB demonstrate significant neurocognitive deficits. Adenotonsillectomy is the first line of treatment for SDB and whilst this improves respiratory disturbance, it remains to be established whether neurocognitive gains also result. METHODS: A total of 44 healthy snoring children aged 3-12 years awaiting adenotonsillectomy (SDB group), and 48 age and gender matched non-snoring controls from the general community, completed the study. All children underwent polysomnography and neurocognitive assessment at baseline and after a 6-month follow-up (after surgery in the snoring group). Our primary aim was to determine whether neurocognitive deficits in snoring children were significantly improved following adenotonsillectomy. RESULTS: Wide ranging neurocognitive deficits were found at baseline in SDB children compared to controls, most notably a 10 point IQ difference (P<.001) and similar deficits in language and executive function. Whilst adenotonsillectomy improved respiratory parameters and snoring frequency at 6 months post surgery, neurocognitive performance did not improve relative to controls. CONCLUSION: Adenotonsillectomy successfully treated the respiratory effects of SDB in children. However, neurocognitive deficits did not improve 6 months post-operatively. PMID- 19806215 TI - The bile acid synthesis pathway is present and functional in the human ovary. AB - BACKGROUND: Bile acids, end products of the pathway for cholesterol elimination, are required for dietary lipid and fat-soluble vitamin absorption and maintain the balance between cholesterol synthesis in the liver and cholesterol excretion. They are composed of a steroid structure and are primarily made in the liver by the oxidation of cholesterol. Cholesterol is also highly abundant in the human ovarian follicle, where it is used in the formation of the sex steroids. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe for the first time evidence that all aspects of the bile acid synthesis pathway are present in the human ovarian follicle, including the enzymes in both the classical and alternative pathways, the nuclear receptors known to regulate the pathway, and the end product bile acids. Furthermore, we provide functional evidence that bile acids are produced by the human follicular granulosa cells in response to cholesterol presence in the culture media. CONCLUSIONS/SIGNIFICANCE: These findings establish a novel pathway present in the human ovarian follicle that has the capacity to compete directly with sex steroid synthesis. PMID- 19806216 TI - High resolution MEMS accelerometers to estimate VO2 and compare running mechanics between highly trained inter-collegiate and untrained runners. AB - BACKGROUND: The purposes of this study were to determine the validity and reliability of high resolution accelerometers (HRA) relative to VO(2) and speed, and compare putative differences in HRA signal between trained (T) and untrained (UT) runners during treadmill locomotion. METHODOLOGY: Runners performed 2 incremental VO(2max) trials while wearing HRA. RMS of high frequency signal from three axes (VT, ML, AP) and the Euclidean resultant (RES) were compared to VO(2) to determine validity and reliability. Additionally, axial rms relative to speed, and ratio of axial accelerations to RES were compared between T and UT to determine if differences in running mechanics could be identified between the two groups. PRINCIPAL FINDINGS: Regression of RES was strongly related to VO(2), but T was different than UT (r = 0.96 vs 0.92; p<.001) for walking and running. During walking, only the ratio of ML and AP to RES were different between groups. For running, nearly all acceleration parameters were lower for T than UT, the exception being ratio of VT to RES, which was higher in T than UT. All of these differences during running were despite higher VO(2), O(2) cost, and lower RER in T vs UT, which resulted in no significant difference in energy expenditure between groups. CONCLUSIONS/SIGNFICANCE: These results indicate that HRA can accurately and reliably estimate VO(2) during treadmill locomotion, but differences exist between T and UT that should be considered when estimating energy expenditure. Differences in running mechanics between T and UT were identified, yet the importance of these differences remains to be determined. PMID- 19806217 TI - rs5888 variant of SCARB1 gene is a possible susceptibility factor for age-related macular degeneration. AB - Major genetic factors for age-related macular degeneration (AMD) have recently been identified as susceptibility risk factors, including variants in the CFH gene and the ARMS2 LOC387715/HTRA1locus. Our purpose was to perform a case control study in two populations among individuals who did not carry risk variants for CFHY402H and LOC387715 A69S (ARMS2), called "study" individuals, in order to identify new genetic risk factors. Based on a candidate gene approach, we analyzed SNP rs5888 of the SCARB1 gene, coding for SRBI, which is involved in the lipid and lutein pathways. This study was conducted in a French series of 1241 AMD patients and 297 controls, and in a North American series of 1257 patients with advanced AMD and 1732 controls. Among these individuals, we identified 61 French patients, 77 French controls, 85 North American patients and 338 North American controls who did not carry the CFH nor ARMS2 polymorphisms. An association between AMD and the SCARB1 gene was seen among the study subjects. The genotypic distribution of the rs5888 polymorphism was significantly different between cases and controls in the French population (p<0.006). Heterozygosity at the rs5888 SNP increased risk of AMD compared to the CC genotypes in the French study population (odds ratio (OR) = 3.5, CI95%: 1.4-8.9, p<0.01) and after pooling the 2 populations (OR = 2.9, 95% CI: 1.6-5.3, p<0.002). Subgroup analysis in exudative forms of AMD revealed a pooled OR of 3.6 for individuals heterozygous for rs5888 (95% CI: 1.7-7.6, p<0.0015). These results suggest the possible contribution of SCARB1, a new genetic factor in AMD, and implicate a role for cholesterol and antioxidant micronutrient (lutein and vitamin E) metabolism in AMD. PMID- 19806218 TI - Coral fluorescent proteins as antioxidants. AB - BACKGROUND: A wide array of fluorescent proteins (FP) is present in anthozoans, although their biochemical characteristics and function in host tissue remain to be determined. Upregulation of FP's frequently occurs in injured or compromised coral tissue, suggesting a potential role of coral FPs in host stress responses. METHODOLOGY/PRINCIPAL FINDINGS: The presence of FPs was determined and quantified for a subsample of seven healthy Caribbean coral species using spectral emission analysis of tissue extracts. FP concentration was correlated with the in vivo antioxidant potential of the tissue extracts by quantifying the hydrogen peroxide (H(2)O(2)) scavenging rates. FPs of the seven species varied in both type and abundance and demonstrated a positive correlation between H(2)O(2) scavenging rate and FP concentration. To validate this data, the H(2)O(2) scavenging rates of four pure scleractinian FPs, cyan (CFP), green (GFP), red (RFP) and chromoprotein (CP), and their mutant counterparts (without chromophores), were investigated. In vitro, each FP scavenged H(2)O(2) with the most efficient being CP followed by equivalent activity of CFP and RFP. Scavenging was significantly higher in all mutant counterparts. CONCLUSIONS/SIGNIFICANCE: Both naturally occurring and pure coral FPs have significant H(2)O(2) scavenging activity. The higher scavenging rate of RFP and the CP in vitro is consistent with observed increases of these specific FPs in areas of compromised coral tissue. However, the greater scavenging ability of the mutant counterparts suggests additional roles of scleractinian FPs, potentially pertaining to their color. This study documents H(2)O(2) scavenging of scleractinian FPs, a novel biochemical characteristic, both in vivo across multiple species and in vitro with purified proteins. These data support a role for FPs in coral stress and immune responses and highlights the multi-functionality of these conspicuous proteins. PMID- 19806219 TI - Simulations of tubulin sheet polymers as possible structural intermediates in microtubule assembly. AB - The microtubule assembly process has been extensively studied, but the underlying molecular mechanism remains poorly understood. The structure of an artificially generated sheet polymer that alternates two types of lateral contacts and that directly converts into microtubules, has been proposed to correspond to the intermediate sheet structure observed during microtubule assembly. We have studied the self-assembly process of GMPCPP tubulins into sheet and microtubule structures using thermodynamic analysis and stochastic simulations. With the novel assumptions that tubulins can laterally interact in two different forms, and allosterically affect neighboring lateral interactions, we can explain existing experimental observations. At low temperature, the allosteric effect results in the observed sheet structure with alternating lateral interactions as the thermodynamically most stable form. At normal microtubule assembly temperature, our work indicates that a class of sheet structures resembling those observed at low temperature is transiently trapped as an intermediate during the assembly process. This work may shed light on the tubulin molecular interactions, and the role of sheet formation during microtubule assembly. PMID- 19806220 TI - Pseudomonas aeruginosa LPS or flagellin are sufficient to activate TLR-dependent signaling in murine alveolar macrophages and airway epithelial cells. AB - BACKGROUND: The human lung is exposed to a large number of airborne pathogens as a result of the daily inhalation of 10,000 liters of air. Innate immunity is thus essential to defend the lungs against these pathogens. This defense is mediated in part through the recognition of specific microbial ligands by Toll-like receptors (TLR) of which there are at least 10 in humans. Pseudomonas aeruginosa is the main pathogen that infects the lungs of cystic fibrosis patients. Based on whole animal experiments, using TLR knockout mice, the control of this bacterium is believed to occur by the recognition of LPS and flagellin by TLRs 2,4 and 5, respectively. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we investigated in vitro the role of these same TLR and ligands, in alveolar macrophage (AM) and epithelial cell (EC) activation. Cellular responses to P. aeruginosa was evaluated by measuring KC, TNF-alpha, IL-6 and G-CSF secretion, four different markers of the innate immune response. AM and EC from WT and TLR2, 4, 5 and MyD88 knockout mice for were stimulated with the wild-type P. aeruginosa or with a mutant devoid of flagellin production. CONCLUSIONS/SIGNIFICANCE: The results clearly demonstrate that only two ligand/receptor pairs are necessary for the induction of KC, TNF-alpha, and IL-6 synthesis by P. aeruginosa-activated cells, i.e. TLR2,4/LPS and TLR5/flagellin. Either ligand/receptor pair is sufficient to sense the bacterium and to trigger cell activation, and when both are missing lung EC and AM are unable to produce such a response as were cells from MyD88(-/-) mice. PMID- 19806221 TI - Hierarchical structure controls nanomechanical properties of vimentin intermediate filaments. AB - Intermediate filaments (IFs), in addition to microtubules and microfilaments, are one of the three major components of the cytoskeleton in eukaryotic cells, playing a vital role in mechanotransduction and in providing mechanical stability to cells. Despite the importance of IF mechanics for cell biology and cell mechanics, the structural basis for their mechanical properties remains unknown. Specifically, our understanding of fundamental filament properties, such as the basis for their great extensibility, stiffening properties, and their exceptional mechanical resilience remains limited. This has prevented us from answering fundamental structure-function relationship questions related to the biomechanical role of intermediate filaments, which is crucial to link structure and function in the protein material's biological context. Here we utilize an atomistic-level model of the human vimentin dimer and tetramer to study their response to mechanical tensile stress, and describe a detailed analysis of the mechanical properties and associated deformation mechanisms. We observe a transition from alpha-helices to beta-sheets with subsequent interdimer sliding under mechanical deformation, which has been inferred previously from experimental results. By upscaling our results we report, for the first time, a quantitative comparison to experimental results of IF nanomechanics, showing good agreement. Through the identification of links between structures and deformation mechanisms at distinct hierarchical levels, we show that the multi-scale structure of IFs is crucial for their characteristic mechanical properties, in particular their ability to undergo severe deformation of approximately 300% strain without breaking, facilitated by a cascaded activation of a distinct deformation mechanisms operating at different levels. This process enables IFs to combine disparate properties such as mechanosensitivity, strength and deformability. Our results enable a new paradigm in studying biological and mechanical properties of IFs from an atomistic perspective, and lay the foundation to understanding how properties of individual protein molecules can have profound effects at larger length-scales. PMID- 19806222 TI - Is there a link between the lipopolysaccharide of Helicobacter pylori gastric MALT lymphoma associated strains and lymphoma pathogenesis? AB - The aim of this study was to investigate the Lewis antigen expression in Helicobacter pylori gastric MALT lymphoma associated strains in comparison to chronic gastritis only strains. Forty MALT strains (19 cagPAI (-) and 21 cagPAI (+)) and 39 cagPAI frequency-matched gastritis strains (17 cagPAI (-) and 22 cagPAI (+)) were included in this study. The lipopolyssacharide for each strain was extracted using a hot phenol method and the expression of Le(x) and Le(y) were investigated using Western Blot. The data were analyzed according to the strains' cagPAI status and vacA genotype. Le(x) was identified in 21 (52.5%) MALT strains and 29 (74.3%) gastritis strains. Le(y) was identified in 30 (75%) MALT strains and 31 (79.5%) gastritis strains. There was an association between cagPAI positivity and Le(x) expression among MALT strains (p<0.0001), but not in gastritis strains (p = 0.64). Among cagPAI (-) strains, isolates expressing solely Le(y) were associated with MALT with an odds ratio of 64.2 (95% CI 4.9 841.0) when compared to strains expressing both Le(x) and Le(y). vacA genotypes did not modify the association between Lewis antigen expression and disease status. In conclusion, cagPAI (-) MALT strains have a particular Lewis antigen profile which could represent an adaptive mechanism to the host response or participate in MALT lymphomagenesis. PMID- 19806223 TI - Measuring morbidity associated with urinary schistosomiasis: assessing levels of excreted urine albumin and urinary tract pathologies. AB - BACKGROUND: Urinary schistosomiasis is responsible for a variety of debilitating conditions; foremost perhaps are urinary tract pathologies (UTPs). Although portable ultrasonography can be used to detect UTPs visually, there is still a need for rapid morbidity assessment (henceforth referred to as RaMA) tools that can be deployed in the field during implementation, monitoring and evaluation of control programmes. We therefore aimed to determine associations between excreted urine-albumin, as measured using a HemoCue photometer, and UTPs, as detected by ultrasonography, in children and adults from an urinary schistosomiasis endemic area in Zanzibar. METHODOLOGY/PRINCIPAL FINDINGS: In a survey of 140 school children of both sexes (aged 9 to 15 yr) and 47 adult males (> or =16 yr) on the island of Unguja, the prevalence of egg-patent urinary schistosomiasis was 36.4% (CI(95) 28.5-45.0%) and 46.8% (CI(95) 32.1-61.9%) (P = 0.14), and that of UTPs was 39.4% (CI(95) 31.0-48.3%) and 64.4% (CI(95) 48.8-78.1%) (P = 0.006), respectively. In school-children, raised urine-albumin concentrations (>40 mg/L) were associated, albeit non-significantly, with prevalence of infection (OR = 3.1, P = 0.070), but more specifically and significantly with the prevalence of micro-haematuria (OR = 76.7, P<0.0001). In adults, elevated urine-albumin excretion was associated with UTPs, particularly lesions of the bladder wall (OR = 8.4, P = 0.013). Albuminuria showed promising diagnostic performance, especially in school-aged children with sensitivity of 63.3% and specificity of 83.1% at detecting lower UTPs, i.e. bladder-wall lesions (ultrasonography as 'gold standard'). CONCLUSION/SIGNIFICANCE: This study indicates that albuminuria assays could be used as a RaMA tool for monitoring UTP prevalence during control programmes, as well as a tool for selecting those with more chronic bladder-wall lesions without resorting to ultrasonography. PMID- 19806224 TI - Transmissibility of atypical scrapie in ovine transgenic mice: major effects of host prion protein expression and donor prion genotype. AB - Atypical scrapie or Nor98 has been identified as a transmissible spongiform encephalopathy (TSE) that is clearly distinguishable from classical scrapie and BSE, notably regarding the biochemical features of the protease-resistant prion protein PrP(res) and the genetic factors involved in susceptibility to the disease. In this study we transmitted the disease from a series of 12 French atypical scrapie isolates in a transgenic mouse model (TgOvPrP4) overexpressing in the brain approximately 0.25, 1.5 or 6x the levels of the PrP(ARQ) ovine prion protein under the control of the neuron-specific enolase promoter. We used an approach based on serum PrP(c) measurements that appeared to reflect the different PrP(c) expression levels in the central nervous system. We found that transmission of atypical scrapie, much more than in classical scrapie or BSE, was strongly influenced by the PrP(c) expression levels of TgOvPrP4 inoculated mice. Whereas TgOvPrP4 mice overexpressing approximately 6x the normal PrP(c) level died after a survival periods of 400 days, those with approximately 1.5x the normal PrP(c) level died at around 700 days. The transmission of atypical scrapie in TgOvPrP4 mouse line was also strongly influenced by the prnp genotypes of the animal source of atypical scrapie. Isolates carrying the AF(141)RQ or AHQ alleles, associated with increased disease susceptibility in the natural host, showed a higher transmissibility in TgOvPrP4 mice. The biochemical analysis of PrP(res) in TgOvPrP4 mouse brains showed a fully conserved pattern, compared to that in the natural host, with three distinct PrP(res) products. Our results throw light on the transmission features of atypical scrapie and suggest that the risk of transmission is intrinsically lower than that of classical scrapie or BSE, especially in relation to the expression level of the prion protein. PMID- 19806225 TI - MicroRNAs show mutually exclusive expression patterns in the brain of adult male rats. AB - BACKGROUND: The brain is a major site of microRNA (miRNA) gene expression, but the spatial expression patterns of miRNAs within the brain have not yet been fully covered. METHODOLOGY/PRINCIPAL FINDINGS: We have characterized the regional expression profiles of miRNAs in five distinct regions of the adult rat brain: amygdala, cerebellum, hippocampus, hypothalamus and substantia nigra. Microarray profiling uncovered 48 miRNAs displaying more than three-fold enrichment between two or more brain regions. Notably, we found reciprocal expression profiles for a subset of the miRNAs predominantly found (> ten times) in either the cerebellum (miR-206 and miR-497) or the forebrain regions (miR-132, miR-212, miR-221 and miR 222). CONCLUSIONS/SIGNIFICANCE: The results indicate that some miRNAs could be important for area-specific functions in the brain. Our data, combined with previous studies in mice, provides additional guidance for future investigations of miRNA functions in the brain. PMID- 19806226 TI - Multivariate statistical analyses demonstrate unique host immune responses to single and dual lentiviral infection. AB - BACKGROUND: Feline immunodeficiency virus (FIV) and human immunodeficiency virus (HIV) are recently identified lentiviruses that cause progressive immune decline and ultimately death in infected cats and humans. It is of great interest to understand how to prevent immune system collapse caused by these lentiviruses. We recently described that disease caused by a virulent FIV strain in cats can be attenuated if animals are first infected with a feline immunodeficiency virus derived from a wild cougar. The detailed temporal tracking of cat immunological parameters in response to two viral infections resulted in high-dimensional datasets containing variables that exhibit strong co-variation. Initial analyses of these complex data using univariate statistical techniques did not account for interactions among immunological response variables and therefore potentially obscured significant effects between infection state and immunological parameters. METHODOLOGY AND PRINCIPAL FINDINGS: Here, we apply a suite of multivariate statistical tools, including Principal Component Analysis, MANOVA and Linear Discriminant Analysis, to temporal immunological data resulting from FIV superinfection in domestic cats. We investigated the co-variation among immunological responses, the differences in immune parameters among four groups of five cats each (uninfected, single and dual infected animals), and the "immune profiles" that discriminate among them over the first four weeks following superinfection. Dual infected cats mount an immune response by 24 days post superinfection that is characterized by elevated levels of CD8 and CD25 cells and increased expression of IL4 and IFNgamma, and FAS. This profile discriminates dual infected cats from cats infected with FIV alone, which show high IL-10 and lower numbers of CD8 and CD25 cells. CONCLUSIONS: Multivariate statistical analyses demonstrate both the dynamic nature of the immune response to FIV single and dual infection and the development of a unique immunological profile in dual infected cats, which are protected from immune decline. PMID- 19806236 TI - Breakthrough in diabetes therapy ... Just around the corner? AB - Inspired by the articles presented in this issue of The Review of Diabetic Studies, we considered it useful to summarize the latest achievements and current challenges we face in the search for a cure of type 1 diabetes. In this editorial article, we took into account how the research landscape has changed in only a few years. While modern lifestyles impose new concerns, now we have a better knowledge of the various aspects of the disease that can be used to treat our young patients with more appropriate approaches, thereby eliminating old and obsolete prejudices. PMID- 19806227 TI - Human sirt-1: molecular modeling and structure-function relationships of an unordered protein. AB - BACKGROUND: Sirt-1 is a NAD+-dependent nuclear deacetylase of 747 residues that in mammals is involved in various important metabolic pathways, such as glucose metabolism and insulin secretion, and often works on many different metabolic substrates as a multifunctional protein. Sirt-1 down-regulates p53 activity, rising lifespan, and cell survival; it also deacetylases peroxisome proliferator activated receptor-gamma (PPAR-gamma) and its coactivator 1 alpha (PGC-1alpha), promoting lipid mobilization, positively regulating insulin secretion, and increasing mitochondrial dimension and number. Therefore, it has been implicated in diseases such as diabetes and the metabolic syndrome and, also, in the mechanisms of longevity induced by calorie restriction. Its whole structure is not yet experimentally determined and the structural features of its allosteric site are unknown, and no information is known about the structural changes determined by the binding of its allosteric effectors. METHODOLOGY: In this study, we modelled the whole three-dimensional structure of Sirt-1 and that of its endogenous activator, the nuclear protein AROS. Moreover, we modelled the Sirt-1/AROS complex in order to study the structural basis of its activation and regulation. CONCLUSIONS: Amazingly, the structural data show that Sirt-1 is an unordered protein with a globular core and two large unordered structural regions at both termini, which play an important role in the protein-protein interaction. Moreover, we have found on Sirt-1 a conserved pharmacophore pocket of which we have discussed the implication. PMID- 19806237 TI - The role of the CXCL10/CXCR3 system in type 1 diabetes. AB - Despite intervention with insulin, type 1 diabetes gradually deteriorates the patients' quality of life. The disease is characterized by an immune-mediated destruction of pancreatic beta-cells. Its etiology, however, remains controversial. Some studies argue that glutamic acid decarboxylase (GAD) antigen and GAD-reactive T cells are critical players in the development of diabetes by affecting the Th cell balance. A T-helper 1 (Th1)-dominant immune response is considered to be important in beta-cell failure in both human and animal models of type 1 diabetes. The Th1-type chemokine, CXCL10, and its receptor, CXCR3, are involved not only in the immune response, but also in the suppression of beta cell proliferation. Thus, understanding the CXCL10/CXCR3 system may be important for finding a cure. In this short review, we discuss the role of the CXCL10/CXCR3 system in type 1 diabetes and propose relevant treatment options. PMID- 19806239 TI - Type 1 diabetes development requires both CD4+ and CD8+ T cells and can be reversed by non-depleting antibodies targeting both T cell populations. AB - Type 1 diabetes development in NOD mice appears to require both CD4(+) and CD8(+) T cells. However, there are some situations where it has been suggested that either CD4(+) or CD8(+) T cells are able to mediate diabetes in the absence of the other population. In the case of transgenic mice, this may reflect the numbers of antigen-specific T cells able to access the pancreas and recruit other cell types such as macrophages leading to a release of high concentrations of damaging cytokines. Previous studies examining the requirement for CD8(+) T cells have used antibodies specific for CD8alpha. It is known that CD8alpha is expressed not only on alphabeta T cells, but also on other cell types, including a DC population that may be critical for presenting islet antigen in the pancreatic draining lymph nodes. Therefore, we have re-examined the need for both CD4(+) and CD8(+) T cell populations in diabetes development in NOD mice using an antibody to CD8beta. Our studies indicate that by using highly purified populations of T cells and antibodies specific for CD8(+) T cells, there is indeed a need for both cell types. In accordance with some other reports, we found that CD4(+) T cells appeared to be able to access the pancreas more readily than CD8(+) T cells. Despite the ability of CD4(+) T cells to recruit CD11b class II positive cells, diabetes did not develop in the absence of CD8(+) T cells. These studies support the observation that CD8(+) T cells may be final effector cells. As both T cell populations are clearly implicated in diabetes development, we have used a combination of non-depleting antibodies to target both CD4 positive and CD8-positive cells and found that this antibody combination was able to reverse diabetes onset in NOD mice as effectively as anti-CD3 antibodies. PMID- 19806238 TI - Histopathology of type 1 diabetes: old paradigms and new insights. AB - Although our knowledge on the various aspects of diabetes development in the NOD mouse model is substantial and keeps expanding at a dramatic pace, the dataset on histopathologic features of type 1 diabetes (T1D) in patients remains largely stagnant. Early work has established an array of common aspects that have become epitomic in the absence of new patient material. There is a growing consensus that an updated and more detailed view is required that challenges and expands our understanding. Comprehensive initiatives are currently ongoing to address these issues in pre-diabetic, recent onset and longstanding type 1 diabetic individuals, and some of the old data have been recently revisited. In this review article, we wish to provide an overview of where we stand today and how we can correlate the various cross-sectional studies from the past with contemporary models of the disease. We believe an enhanced understanding of the many histopathological particularities in patients as compared to animal models will ultimately lead, not only to more fundamental insights, but also to an improved ability to translate pre-clinical data from bench to bedside. PMID- 19806240 TI - Allelic variation of Ets1 does not contribute to NK and NKT cell deficiencies in type 1 diabetes susceptible NOD mice. AB - The NOD mouse is a well characterized model of type 1 diabetes that shares several of the characteristics of Ets1-deficient targeted mutant mice, viz: defects in TCR allelic exclusion, susceptibility to a lupus like disease characterized by IgM and IgG autoantibodies and immune complex-mediated glomerulonephritis, and deficiencies of NK and NKT cells. Here, we sought evidence for allelic variation of Ets1 in mice contributing to the NK and NKT cell phenotypes of the NOD strain. ETS1 expression in NK and NKT cells was reduced in NOD mice, compared to C57BL/6 mice. Although NKT cells numbers were significantly correlated with ETS1 expression in both strains, NKT cell numbers were not linked to the Ets1 gene in a first backcross from NOD to C57BL/6 mice. These results indicate that allelic variation of Ets1 did not contribute to variation in NKT cell numbers in these mice. It remains possible that a third factor not linked to the Ets1 locus controls both ETS1 expression and subsequently NK and NKT cell phenotypes. PMID- 19806241 TI - Comparison of fasting glucose with post-load glucose values and glycated hemoglobin for prediction of type 2 diabetes: the Isfahan diabetes prevention study. AB - OBJECTIVES: The aim of this study was to compare the ability of fasting plasma glucose (FPG), post-load plasma glucose values and glycated hemoglobin (HbA1c) to predict progression to diabetes in non-diabetic first-degree relatives (FDR) of patients with type 2 diabetes. METHODS: A total of 701 non-diabetic FDR of diabetic patients aged 20-70 years surveyed in 2003 to 2005 were followed until 2008 for the onset of type 2 diabetes mellitus. At baseline and at follow-ups, participants underwent a standard 75 g 2-hour oral glucose tolerance test (OGTT). Prediction of progression to type 2 diabetes was assessed by using area under the receiver-operating characteristic (ROC) curves based upon measurement of FPG, post-load glucose values and HbA1c. RESULTS: The incidence of type 2 diabetes was 33.9 per 1000 person-years in men and 48.6 in women. The incidence rates were 4.6, 50.7, and 99.7 per 1000 person-years in FDR with normal glucose tolerance, impaired fasting glucose and impaired glucose tolerance respectively. FPG value was a better predictor of progression to diabetes than any post-load glucose values or HbA1c. The areas under the ROC curves were 0.811 for fasting, 0.752 for 1/2-hour, 0.782 for 1-hour and 0.756 for 2-hour glucose vs. 0.634 for HbA1c (p < 0.001). CONCLUSIONS: FPG had more discriminatory power to distinguish between individuals at risk for diabetes and those who were not at risk than post-load glucose values during OGTT or HbA1c. Our findings support the American Diabetes Association recommendation of using FPG concentration to diagnose diabetes. PMID- 19806242 TI - Prevalence of diabetic retinopathy and cataract in adult patients with type 1 and type 2 diabetes in Russia. AB - AIM: The aim of the study was to identify the prevalence of diabetic retinopathy (DR) and diabetic cataract (DC) in type 1 and type 2 diabetic patients within the Russian Federation. Also, the stage of DR at the time of its identification and the proportion of new cases diagnosed with DR or DC were to be determined. METHODS: A random sample of 7,186 adult patients with diabetes was screened for DR and DC using fundoscopy and fundus photography. Levels of HbA1c, total cholesterol, triglycerides, creatinine and urinary albumin excretion rate were assessed. RESULTS: In diabetic patients, the prevalence of DR and DC was 45.9% and 30.6%, respectively. These complications appeared significantly more frequently in patients with type 1 diabetes than in type 2 diabetes. The prevalence of background, preproliferative and proliferative DR among diabetic patients was 28.1%, 8.1%, and 6.7%, respectively. Patients with DR were older, had a longer duration of diabetes, higher HbA1c, elevated plasma total cholesterol, increased triglicerides, and higher systolic BP, compared with patients without DR. Microalbuminuria and proteinuria were more prevalent among patients with DR compared with non-DR patients. CONCLUSIONS: The results showed that diabetic retinopathy and cataract are wide-spread complications among diabetic patients in Russia. However, the disease course is more aggressive and accelerated in patients with type 1 diabetes than in those having type 2 diabetes. Therefore, it is important to prevent DR by identifying diabetes and signs of retinopathy at the earliest possible stage of progression for timely and adequate retina laser coagulation or surgical treatment, compensation of carbohydrate and lipid metabolism, and normalization of blood glucose and pressure. PMID- 19806243 TI - Does weather influence the development of deep venous thrombosis? The answer may be blowing in the wind. PMID- 19806244 TI - Platelets interact with bacterial pathogens. PMID- 19806246 TI - Risk factors for cerebral venous thrombosis and deep venous thrombosis in patients aged between 15 and 50 years. AB - Cerebral venous thrombosis (CVT) and deep vein thrombosis or pulmonary embolism (DVT/PE) are associated with many risk factors. It is unclear why CVT occurs less often than DVT/PE. Age dependent risk factors may play a role. The aim of our study was to compare risk factors in a uniform age group of CVT and DVT/PE patients aged between 15 and 50 years. Thrombophilic markers and clinical risk factors of 79 CVT patients and 173 DVT/PE patients aged 15-50 years were compared. Multivariable logistic regression analysis was performed to investigate if risk factors were independently associated with CVT or DVT/PE. Cerebral venous thrombosis patients were younger (median age 30 years vs. 42 years; p<0.001) and more often female (82% vs. 52%; p<0.001). There were no differences in thrombophilic markers. Cerebral venous thrombosis was less often associated with trauma, immobilisation or surgery than DVT/PE (6% vs. 21%; adjusted OR 0.29; 95%CI 0.10-0.82). In women, CVT was more frequently associated with oral contraceptive use, pregnancy or puerperium (82% vs. 53%; adjusted OR 2.34; 95%CI 1.03-5.32). This study demonstrated no differences in thrombophilic markers between CVT patients and DVT/PE patients aged between 15 and 50 years, while the frequency of some transient risk factors was different. Cerebral venous thrombosis was relatively more common in women and hormonal factors may predispose to CVT compared to DVT/PE, while trauma, immobilisation and surgery may be less important in the pathophysiology of CVT. PMID- 19806245 TI - C-reactive protein and venous thromboembolism. A prospective investigation in the ARIC cohort. AB - The role of inflammation in the causation of venous thromboembolism (VTE) is uncertain. In 10,505 participants of the Atherosclerosis Risk in Communities (ARIC) Study, we assessed the association of the systemic inflammation marker, elevated C-reactive protein (CRP), with incidence of VTE (n=221) over a median of 8.3 years of follow-up. Adjusted for age, race, and sex, the hazard ratios of VTE across quintiles of CRP were 1.0, 1.61, 1.16, 1.56, and 2.31 (p for trend p<0.0007). For CRP above the upper 10 percentile (> or = 8.55 mg/L), compared with the lowest 90% of CRP values, the hazard ratio of VTE was 2.07 (95% CI 1.47, 2.94). Further adjustment for baseline hormone replacement therapy, diabetes, and body mass index attenuated the hazard ratios only slightly. For example, the adjusted hazard ratio of VTE was 1.76 (95% CI 1.23, 2.52) for CRP above versus below the 90(th) percentile. In conclusion, this prospective, population-based study suggests elevated CRP is independently associated with increased risk of VTE. PMID- 19806247 TI - Differences in prethrombin-1 activation with human or bovine factor Va can be attributed to the heavy chain. AB - Human and bovine factor Va (FVa) function similarly in the activation of prothrombin but differently in the activation of prethrombin-1 (Pre-1). Pre-1 activation with human FVa proceeds at about 22 percent of the rate with bovine FVa. The dependencies of initial rates on the FVa and Pre-1 concentrations indicate that the differential activity is expressed in kcat differences, rather than differences in the assembly of prothrombinase or the K(m) value of the substrate. The heavy and light chains of both species of FVa were separated and interspecies hybrids were constructed in the presence of Ca(++). Studies of the activation of Pre-1 with these hybrids indicate that the species difference can be attributed specifically to the heavy chain of FVa. Analyses of the reactions by SDS-PAGE indicated that cleavage at Arg271 occurs at about the same rate with both species of FVa, but cleavage at Arg320 with human FVa is specifically retarded. A major difference in primary structure between the human and bovine FVa heavy chains comprises 10 residues at COOH-terminus, adjacent to the negatively charged hirudin-like DYDYQ sequence. These residues have pI values of 12.5 and 4.26 in human and bovine FVa, respectively. The lower value would complement the negatively charged DYDYQ sequence but the higher value would counteract it. Thus, we suggest that the differences in the COOH-terminus of the heavy chain are responsible for the differences in Pre-1 activation, and that it specifically influences cleavage at Arg320 in Pre-1. PMID- 19806248 TI - Genetic fusion to albumin improves the pharmacokinetic properties of factor IX. AB - Haemophilia B is a X-chromosome linked disease characterised by a deficiency of functionally active coagulation Factor IX (FIX). Patients with severe haemophilia B at risk of recurrent bleeding are treated approximately twice a week in a prophylactic setting by application of FIX concentrates. To increase convenience and compliance of the therapy it is desirable to reduce the dosing frequency by improving the pharmacokinetic properties of FIX. Here a concept of rFIX (recombinant factor IX) albumin fusion proteins (rIX-FPs) with cleavable linker peptides derived from the FIX activation sequence is presented. Constructs of the genetic fusion of FIX to albumin via cleavable linkers were expressed in mammalian cells and characterised after purification. In vitro activation studies with FXIa demonstrated that cleavage of the linker and the activation peptide proceeded comparably well. In a clotting assay the rIX-FPs with cleavable linker showed a 10- to 30-fold increase in the molar specific clotting activity compared to fusion proteins with non-cleavable linkers. Furthermore, in-vivo recovery, terminal half-life and the AUC of rIX-FPs in rats and rabbits as determined by FIX antigen measurements were significantly increased compared to rFIX (BeneFIX). In FIX deficient (FIX(-/-)) mice the in-vivo recovery and the AUC were also significantly increased. The efficacy in reducing bleeding time was shown in FIX( /-) mice by a tail tip bleeding model. The results suggest that rIX-FPs with a cleavable linker between FIX and albumin are a promising concept that may support the use of the albumin fusion technology to extend the half-life of FIX. PMID- 19806249 TI - Identification of a protein S-interactive site within the A2 domain of the factor VIII heavy chain. AB - We have recently demonstrated that protein S impairs the intrinsic tenase complex, independent of activated protein C, in competitive interactions between the A2 and A3 domains of factor VIIIa and factor IXa. In the present study, we have identified a protein S-interactive site in the A2 domain of factor VIIIa. Anti-A2 monoclonal antibody recognising a factor IXa-functional region (residues 484-509) on A2, and synthetic peptide inhibited the A2 binding to protein S by approximately 60% and approximately 70%, respectively, in solid-phase binding assays. The 484-509 peptide directly bound to protein S dose-dependently. Covalent cross-linking was observed between the 484-509 peptide and protein S following reaction with EDC (1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide). The cross-linked adduct was consistent with 1:1 stoichiometry of reactants. Cross linking formation was blocked by addition of the 484-497 peptide, but not by the 498-509 peptide. Furthermore, N-terminal sequence analysis of the 484-509 peptide protein S adduct showed that three sequential residues (S488, R489, and R490) in A2 were not identified, suggesting that these residues participate in cross-link formation. Mutant A2 molecules where these residues were converted to alanine were evaluated for the binding of protein S. The S488A, R489A, and R490A mutants demonstrated approximately four-fold lower affinity than wild-type A2. These results indicate that the 484-509 region in the A2 domain of factor VIIIa, in particular sequential residues at positions 488-490, contributes to a unique protein S-interactive site. PMID- 19806250 TI - The A6936G polymorphism of the endothelial protein C receptor gene is associated with the risk of unexplained foetal loss in Mediterranean European couples. AB - The endothelial protein C receptor (EPCR) is expressed by trophoblast cells. Mid gestation pregnancy loss is described in animals with a haemochorial placenta lacking EPCR. The A6936G allele of the EPCR gene (PROCR) may be associated with lower EPCR densities on trophoblasts, but data are lacking for its effect on the risk of pregnancy loss in humans. A 1:2 case-control study on unexplained pregnancy loss was nested in the NOHA First cohort: 3,218 case couples and 6,436 control couples were studied for PROCR A6936G, coagulation factor V gene (F5) G1691A and coagulation factor II gene (F2) G20210A polymorphisms. Ethnicity and time of pregnancy loss defined through biometry-based gestational ages (embryonic loss < 10(th) week > or = foetal loss) were analysed. The PROCR A6936G allele, in mothers and fathers, was associated only with foetal loss in both Europeans and non-Europeans. Increasing probability levels of carrying a homozygous child were increasingly associated with the risk of foetal demise. The F5 G1691A and F2 G20210A alleles, only in mothers, were only and independently associated with foetal loss in Europeans. In our population, the PROCR A6936G allele describes women, but also men and thus couples, at risk for first unexplained foetal loss. This risk is independent of the foetal loss risk conferred to our local Mediterranean European women by the F5 G1691A and F2 G20210A alleles. Data confirm that the relationship between thrombophilias and pregnancy loss varies according to ethnicity and loss type. PMID- 19806251 TI - Cardiovascular risk factors in idiopathic compared to risk-associated venous thromboembolism: A focus on fibrinogen, factor VIII, and high-sensitivity C reactive protein (hs-CRP). AB - There have recently been reports of an increased incidence of arterial cardiovascular events in patients with idiopathic venous thromboembolism (VTE) compared to patients with risk-associated VTE. The aim of our study was to evaluate whether elevated clotting factors, which have been linked to chronic sub clinical inflammation and arterial thromboembolic disease, have a higher prevalence in idiopathic VTE compared to secondary VTE. Plasma fibrinogen, factor VIII, and high-sensitivity C-reactive protein (hs-CRP) levels were determined in a cohort of sex- and age-matched patients with unprovoked VTE (n=101), patients with secondary VTE (n=101), and controls (n=202). Fibrinogen and hs-CRP levels were higher in patients with idiopathic VTE (fibrinogen: median/range: 331/214 524 mg/dl; hs-CRP: median/interquartile range: 1.8/0.8-3.7 mg/l) than in those with risk-associated VTE (299/162-458 mg/dl, p=0.004; 1.5/0.8-2.2 mg/l, p=0.05) and controls (302/185-644 mg/dl, p=0.001; 1.2/0.5-2.2 mg/l, p=0.02). Fibrinogen levels in the upper tertile of the controls were seen in 53% of patients with unprovoked VTE, compared to 35% of patients with secondary VTE. According to their hs-CRP levels (>3 mg/l), 26% of patients with idiopathic VTE were categorised as being at high risk for cardiovascular disease, as opposed to just 9% of those with risk-associated VTE. Factor VIII activity was significantly higher in patients with both idiopathic and secondary VTE than in controls, with the highest median value in patients with idiopathic VTE. Our data show that markers of inflammation, such as hs-CRP, fibrinogen, and factor VIII, are at higher levels in patients with idiopathic compared to secondary VTE, supporting the hypothesis that idiopathic VTE and arterial thromboembolism share common risk factors. PMID- 19806252 TI - The influence of meteorological variables on the development of deep venous thrombosis. AB - The influence of weather on deep venous thrombosis (DVT) incidence remains controversial. We aimed to characterize the temporal association between DVT and meteorological variables including atmospheric pressure. Data relating to hospital admissions with DVT in Scotland were collected retrospectively for a 20 year period for which corresponding meteorological recordings were available. Weather variables were calculated as weighted daily averages to adjust for variations in population density. Seasonal variation in DVT and short-term effects of weather variables on the relative risk of developing DVT were assess using Poisson regression modelling. The models allowed for the identification of lag periods between variation in the weather and DVT presentation. A total of 37,336 cases of DVT were recorded. There was significant seasonal variation in DVT with a winter peak. Seasonal variation in wind speed and temperature were significantly associated with seasonal variation in DVT. When studying more immediate meteorological influences, low atmospheric pressure, high wind speed and high rainfall were significantly associated with an increased risk of DVT approximately 9-10 days later. The effect was most strikingly demonstrated for atmospheric pressure, every 10 millibar decrease in pressure being associated with a 2.1% increase in relative risk of DVT. Alterations in weather have a small but significant impact upon the incidence of DVT. DVT is particularly associated with reduction in atmospheric pressure giving weight to the hypothesis that reduced cabin pressure in long haul flights contributes to DVT. These findings have implications for our understanding of the pathogenesis of DVT. PMID- 19806253 TI - Risk stratification of patients with pulmonary embolism based on pulse rate and D dimer concentration. AB - To enable outpatient treatment of a selected group of patients with pulmonary embolism (PE), insight in the determinants of adverse clinical outcome is warranted. We have identified risk factors for serious adverse events (SAE) within the first 10 days of acute PE. We have retrospectively analysed data of 440 consecutive patients with acute PE. Collected data included age, gender, medical history, blood pressure, pulse rate and D-dimer concentration. The variables associated with SAE in the first 10 days in univariate analysis (p<0.15) have been included in a multivariate logistic regression model (backward conditional, p out >0.10). In 440 patients with acute PE, 20 SAEs occurred in a 10-day follow-up period. Pulse rate > or = 100 beats per minute (bpm) (OR, 6.85; 95%CI 1.43-32.81) and D-dimer concentration > or = 3,000 microg/ml (OR, 5.51; 95%CI 0.68-44.64) were significantly related to the SAEs. All SAEs were predicted by a pulse rate > or = 100 bpm and/or a D-dimer concentration > or = 3,000 microg/ml. Older age, gender, history of venous thromboembolism (VTE), heart failure, chronic obstructive pulmonary disease, cancer or a systolic blood pressure < 90 mm Hg had no significant influence on short term SAEs. Pulse rate and D-dimer concentration can be used to identify patients with acute PE, who are at risk for adverse clinical outcome during the first 10 days of hospitalisation. Outpatient treatment of PE-patients with a pulse rate > or = 100 bpm and/or a D dimer concentration > or = 3,000 microg/ml has to be discouraged. PMID- 19806254 TI - Long-term complications of medical patients with hospital-acquired venous thromboembolism. AB - Long-term complications from hospital-acquired acute venous thromboembolism (VTE) include recurrent VTE, postthrombotic syndrome (PTS), and chronic thromboembolic pulmonary hypertension (CTEPH). We used a probability model to estimate the number of these events among hospitalised medical patients in the 2003 United States Healthcare Cost and Utilization Project (HCUP) Nationwide Inpatient Sample database. Of 8,077,919 hospitalised medical patients at risk for VTE, we calculate that 122,235 were stricken with deep vein thrombosis (DVT) and 32,654 with pulmonary embolism (PE). These events generated 49,843 VTE-related deaths, 28,052 recurrent DVTs, 6,680 recurrent PEs, 140,156 cases of PTS, and 5,288 cases of CTEPH over the ensuing 5 years, for a total of 180,176 patients afflicted with long-term complications of VTE. In our model, rates of pharmacological thromboprophylaxis prescribing varied across populations, ranging from 15.3% to 49.2%. When we modeled universal utilisation of pharmacological prophylaxis, the number of VTE-related deaths decreased from 49,843 to 20,739, recurrent DVT was reduced from 28,052 to 13,384, and recurrent PE was reduced from 6,680 to 3,187 events. Incident cases of PTS decreased from 140,156 to 54,651, and CTEPH decreased from 5,288 to 1,115 cases. The number of hospitalised medical patients with long-term VTE complications was reduced by 60% to 72,337. In conclusion, hospitalised medical patients are particularly vulnerable to the development of recurrent VTE, PTS, and CTEPH. These VTE complications would be reduced by more than half with universal thromboprophylaxis. Further efforts should focus on improving VTE prophylaxis utilisation. PMID- 19806255 TI - alpha(1)-acid glycoprotein (AGP)-induced platelet shape change involves the Rho/Rho kinase signalling pathway. AB - alpha(1)-acid glycoprotein (AGP) is an acute-phase protein that contributes to inflammation processes. The role of AGP in platelet activation and thrombosis is, however, largely unknown. Therefore, we thoroughly investigated the effects of AGP on human platelets. Platelets were isolated from healthy volunteers and subsequently exposed to AGP. Platelet responses were monitored as change in light transmission, intracellular calcium concentration, light microscopy and protein phosphorylation by Western blot. We found that AGP induced platelet shape change independently of a second release of adenine nucleotides or thromboxane A(2), and that effect was abolished by endothelium-derived platelet inhibitors such as nitric oxide (NO) and adenosine. Furthermore, AGP triggered a minor calcium response and a pronounced Rho/Rho-kinase-dependent increase in Thr696 phosphorylation of myosin phosphatase target subunit 1 (MYPT1). Moreover, the Rho/Rho-kinase inhibitor Y-27632 significantly decreased the AGP-induced shape change. The results also showed that the AGP-elicited shape change was antagonised by pretreatment with low doses of collagen and thrombospondin-1. Our results describe a novel mechanism by which AGP stimulates platelet shape change via activation of the Rho/Rho-kinase signalling pathway. Physiological important platelet inhibitors, such as NO, completely counterbalance the effect of AGP. Hence, the present study indicates that AGP directly contributes to platelet activation, which in turn might have an impact in physiological haemostasis and/or pathological thrombosis. PMID- 19806256 TI - The tumour necrosis factor superfamily ligand APRIL (TNFSF13) is released upon platelet activation and expressed in atherosclerosis. AB - Activated platelets release a wide range of inflammatory mediators, including members of the tumour necrosis factor (TNF) superfamily (e.g. CD40 ligand [CD40L] and LIGHT). Such platelet-mediated inflammation could be involved in atherogenesis and plaque destabilisation. In the present study we investigated whether APRIL, another member of the TNF superfamily that has been detected in megakaryocytes, could be released from platelets upon activation. The release of APRIL was studied in thrombin receptor (SFLLRN) activated platelets, and the expression of APRIL was examined in plasma and within the atherosclerotic lesion in patients with carotid and coronary atherosclerosis. Upon SFLLRN activation, there was a gradual release of APRIL, reaching maximum after 90 minutes. While this pattern is similar to that of CD40L and LIGHT, the release of APRIL was quite differently regulated. Thus, prostaglandin E1, but not inhibitors of metal dependent proteases and actin polymerisation or the lack of GP IIb/IIIa, blocks APRIL release in activated platelets. With relevance to atherogenesis, we found that patients with coronary artery disease (n=80) had raised plasma levels of APRIL as compared with controls (n=20), and APRIL immunoreactivity was detected in aggregated platelets within the ruptured plaque in patients with myocardial infarction and within macrophages in symptomatic carotid plaques. In conclusion, activated platelets release significant amounts of APRIL in a long-lasting manner, differently regulated than the gradual release of other platelet-derived TNF superfamily ligands. The enhanced expression of APRIL in atherosclerotic disorders, both systemically and within the lesion, may suggest a potential involvement of APRIL in atherogenesis. PMID- 19806257 TI - Proteomic and functional characterisation of platelet microparticle size classes. AB - Activated platelets release large lipid-protein complexes termed microparticles. These platelet microparticles (PMP) are composed of vesicular fragments of the plasma membrane and alpha-granules. PMP facilitate coagulation, promote platelet and leukocyte adhesion to the subendothelial matrix, support angiogenesis and stimulate vascular smooth muscle proliferation. OBJECTIVES: PMP were separated into 4 size classes to facilitate identification of active protein and lipid components. PMP were obtained from activated human platelets and separated into 4 size classes by gel filtration chromatography. Proteins were identified using 2 dimensional, liquid chromatography tandem mass spectrometry. Functional effects on platelets were determined using the PFA-100 and on endothelial cells by measuring transendothelial cell electrical resistance. PMP size classes differed significantly in their contents of plasma membrane receptors and adhesion molecules, chemokines, growth factors and protease inhibitors. The two smallest size classes (3 and 4) inhibited collagen/adenosine-diphosphate-mediated platelet thrombus formation, while fractions 2 and 4 stimulated barrier formation by endothelial cells. Heat denaturation blocked the effect of fraction 4 on endothelial cell function, but not fraction 2 implying that the active component in fraction 4 is a protein and in fraction 2 is a heat-stable protein or lipid but not sphingosine-1-phosphate. Proteomic and functional analysis of PMP size fractions has shown that PMP can be separated into different size classes that differ in protein components, protein/lipid ratio, and functional effects on platelets and endothelial cells. This analysis will facilitate identification of active components in the PMP and clarify their involvement in diseases such as atherosclerosis and cancer. PMID- 19806258 TI - The influence of clinical characteristics, laboratory and inflammatory markers on 'high on-treatment platelet reactivity' as measured with different platelet function tests. AB - High on-clopidogrel platelet reactivity (HCPR) and high on-aspirin platelet reactivity (HAPR) are independently associated with an increased risk of atherothrombotic events. However, despite this positive correlation, the definitions of both HCPR and HAPR vary largely throughout studies and between different platelet function assays. The aim of the present study was to explore clinical and laboratory parameters that are associated with HCPR and HAPR as measured with different platelet function tests. 530 clopidogrel and aspirin pre treated patients undergoing elective PCI (percutaneous coronary intervention) were enrolled. Platelet function measurements were performed with: optical aggregometry, the VerifyNow device and PFA-100 cartridges (including the novel INNOVANCE P2Y assay). HCPR as measured with Adenosin-Di-Phospate-induced (ADP) aggregation based tests was associated with clinical factors such as older age, female gender and Diabetes mellitus (DM). The VerifyNow P2Y12 assay was significantly influenced by haemoglobin and haematocrit levels. HAPR as measured with aggregation based tests was significantly influenced by the presence of malignancy, BMI (Body-Mass Index), older age and increased levels of hsCRP (high sensitivity c-reactive proteine). The PFA-100 COL/EPI (collagen-epinephrine) and COL/ADP (collagen-ADP) cartridges were significantly influenced by monocyte count, hs-CRP, MPV (mean platelet volume), vWF-antigen (von Willebrand factor) and vWF-activity. HCPR as measured with the novel INNOVANCE P2Y cartridge was associated with clinical determinants such as BMI, female gender, impaired LVEF (left ventricular ejection fraction), renal failure and dosing of clopidogrel. Laboratory markers that were associated with HCPR as measured with INNOVANCE P2Y were platelet count, white blood cells (WBC), hsCRP and fibrinogen. Both HCPR and HAPR are highly dependent on the type of platelet function assay. Each platelet function assay, in turn, is significantly influenced by distinct clinical and laboratory variables. PMID- 19806259 TI - Tissue factor activity of blood mononuclear cells is increased after total knee arthroplasty. AB - Tissue factor (TF) is present in small quantities in normal blood and is reported to be elevated in arterial and venous thrombosis. Patients undergoing total knee arthoplasty (TKA) are at high risk of post-operative venous thromboembolism (VTE). To evaluate the possible contribution of elevated blood TF to VTE risk, we performed serial studies of peripheral blood mononuclear cell (PBMC) functional TF procoagulant activity (PCA) in 19 patients after TKA. PBMC and platelet TF PCA were measured by a functional, clot-based assay following decryption with a calcium ionophore. Plasma TF antigen levels were measured by ELISA. All subjects received chemoprophylaxis and none had VTE. After TKA total TF PCA of PBMC was elevated in 19 of 19 subjects. The peak increase above preoperative levels was 1.1-13.6 fold (>two-fold in 58% and >three-fold in 42%). Median TF PCA of PBMC was not elevated following tourniquet removal, but it was significantly elevated on postoperative days 1 and 2. Thereafter, it decreased to near preoperative values at day 6. Neither platelet TF PCA nor plasma TF antigen levels increased significantly. Since the PBMC count did not rise, the increase in TF PCA was attributable to cell synthesis. The increase in blood TF PCA preceded the median time of diagnosis of venous thromboembolism after TKA established previously. These observations indicate a) TKA stimulates synthesis of encrypted PBMC TF PCA which is likely to contribute to the pathophysiology of VTE; b) TF antigen is not a reliable indicator of TF PCA. PMID- 19806260 TI - Pneumococcal association to platelets is mediated by soluble fibrin and supported by thrombospondin-1. AB - Platelets and coagulation are involved in bacterial colonisation of the host. Streptocococcus pneumoniae (pneumococcus) are important etiologic agents of respiratory tract infections in humans. The formation of pneumococci-platelet associations may facilitate haematogenous dissemination of pneumococci by providing an adhesive surface on damaged endothelium. However, the formation of platelet-pneumococci associations and the factors involved in this process have not been described so far. The formation of platelet-pneumococci associates was analysed and quantified using flow cytometry. Binding of pneumococci to platelets was significantly increased after activation of platelets with thrombin, while platelet activation by ADP or collagen did not promote formation of platelet pneumococci associates. In addition to be a platelet agonist, thrombin cleaves fibrinogen, which results in the generation of fibrin. The simultaneous formation of fibrin and activation of platelets was shown to be a prerequisite for a high number of platelet-pneumococci associates. Moreover, exogenously added human thrombospondin-1 (TSP-1) significantly enhanced the association of pneumococci with activated platelets. Soluble fibrin and TSP-1 are key co-factors of platelet pneumococci-association. Similar results were recently demonstrated for S. aureus platelet adhesion. Consequently, we hypothesise that the described mechanism of platelet-bacteria-association might represent a general and important strategy of Gram-positive bacteria during development of invasive diseases. PMID- 19806261 TI - Proliferative stimulation of the vascular Endothelin-1 axis in vitro and ex vivo by infection with Chlamydia pneumoniae. AB - Endothelin-1 (ET-1) is a vasoactive peptide that modifies vascular function via the G-protein coupled transmembrane receptors, Endothelin-A receptor (ETAR) and Endothelin-B receptor (ETBR). Dysregulation of the ET-1 axis plays a role in atherosclerotic development as it triggers cell proliferation, inflammation, and vasoconstriction. The respiratory pathogen Chlamydia pneumoniae (Cp) has been recovered from atherosclerotic lesions, and related to atherogenesis, via activation of vascular small GTPases and leukocyte recruitment. Cp effectively reprograms host cell signalling and is able to enter an intracellular persistent state in vascular cells that is refractory to antibiotics. Upon chlamydial infection, vascular smooth muscle cells, which do not produce significant ET-1 under physiological conditions were switched into a fundamental source of ET-1 mRNA and protein in a p38-MAP-kinase-dependent pathway. Endothelial cells did not overproduce ET-1 but showed upregulation of mitogenic ETAR mRNA and protein while the counterbalancing ETBR, which regulates ET-1 clearance, remained unaffected. This disruption of the ET-1 axis was confirmed in an ex vivo mouse aortic ring model, and resulted in endothelial cell proliferation that could be abrogated by ETAR-siRNA and the selective ETAR-antagonist BQ-123. Chronic chlamydial infection of the vascular wall might represent a permanent noxious stimulus linked to the endothelial cell proliferation characteristic of early atherosclerosis. Suppression of this deleterious paracrine loop by ETAR antagonism opens up a new option of preventing possible vascular sequelae of otherwise untreatable chronic chlamydial infection. In conclusion, this is the first study to demonstrate infection to dysregulate the ET-1 axis towards inducing a proatherogenic proliferative phenotype. PMID- 19806262 TI - Real-life anticoagulation treatment of atrial fibrillation after catheter ablation: Possible overtreatment of low-risk patients. AB - Catheter ablation provides curative treatment for atrial fibrillation (AF). Data on anticoagulation after the procedure are sparse. We investigated real-life antithrombotic treatment after AF ablation and examined its adherence to current recommendations. Eight hundred forty-four patients (age 58 + or - 10 years) underwent AF ablation. Most patients had a CHADS(2) score of 0 (46%) or 1 (45%). Seven-day Holter was performed at three, six and 12 months after ablation. Decision on anticoagulation treatment was made by general practitioners and referring cardiologists in consultation with the patients. At discharge, anticoagulants were prescribed for the vast majority (94-96%) of patients. This percentage remained high at three and six months (80-90%) without differences between stroke risk groups. At 12 months, the use of anticoagulants was mainly influenced by the detection of recurrence; usage exceeded 90% in all stroke risk groups in patients with recurrences. In patients without recurrences, differences between risk groups were significant but small, ranging from 42% (CHADS(2)=0) to 62% (CHADS(2) > or = 2) (p=0.033). In multivariate analysis, the only factor independently associated with oral anticoagulation at 12 months was the detection of recurrences (odds ratio=16.2, p<0.001), whereas the effect of the CHADS(2) score was not significant (p=0.080). The effect of all other examined factors was also not significant. Contrary to current recommendations, anticoagulation after AF ablation is hardly guided by the stroke risk profile and remains high even in low-risk patients. The most important factor influencing the use of anticoagulants is the detection of recurrences during follow-up. This results in possible overtreatment of low-risk patients. PMID- 19806263 TI - Lower concentrations of thrombin-antithrombin complex (TAT) correlate to higher recanalisation rates among ischaemic stroke patients treated with t-PA. AB - An elevated concentration of the thrombin-antithrombin complex (TAT) has been associated with high mortality rates and poor outcome in ischaemic stroke patients treated with tissue plasminogen activator (t-PA). Moreover, antithrombin drugs have been tested in combination with t-PA in the acute phase of ischaemic stroke to increase treatment efficacy. We aimed to study whether poor outcome associated with TAT among ischaemic stroke patients treated with t-PA could be due to the effects of this complex on recanalisation rates of the middle cerebral artery (MCA) and on haemorrhagic transformation. The TAT levels of 89 patients having a proximal MCA occlusion were measured by ELISA, and the patients were then treated with t-PA. Complete recanalisation was diagnosed by transcranial Doppler (TCD) at 1, 2 and 6 hours post-t-PA infusion and haemorrhagic transformation was identified by computed tomography (CT). Lower levels of TAT were associated with better recanalisation rates at all time-points (1 hour: OR = 24.8 95% CI 1.4-434.8, p = 0.028; 2 hours: OR = 6.3 95% CI 1.5-27, p = 0.014; 6 hours: OR = 6.4 95% CI 1.5-26.5, p = 0.011) after adjustment for stroke risk factors. However, no correlation was found between TAT concentration and haemorrhagic transformation. The elevated mortality rates previously observed in patients with high levels of TAT might have been due to revascularisation resistance. Low levels of TAT are not associated with an increase in haemorrhagic complications after t-PA, indicating that the combination of thrombin blockers and t-PA could be a safe and effective treatment for ischaemic stroke in the future. PMID- 19806264 TI - Repletion of factor XIII following cardiopulmonary bypass using a recombinant A subunit homodimer. A preliminary report. AB - Bleeding following cardiac surgery involving cardiopulmonary bypass (CPB) remains a major concern. Coagulation factor XIII (FXIII) functions as a clot-stabilising factor by cross-linking fibrin. Low post-operative levels of FXIII correlate with increased post-operative blood loss. To evaluate preliminary safety and pharmacokinetics of recombinant FXIII (rFXIII-A(2)) in cardiac surgery, patients scheduled for coronary artery bypass grafting were randomised to receive a single dose of either rFXIII-A(2) (11.9, 25, 35 or 50 IU/kg) or placebo in a 4:1 ratio. Study drug was given post-CPB within 10 to 20 minutes after first protamine dose. Patients were evaluated until day 7 or discharge, with a follow-up visit at weeks 5-7. The primary end-point was incidence and severity of adverse events. Thirty five patients were randomised to rFXIII-A(2) and eight to placebo. Eighteen serious adverse events were reported. These were all complications well recognised during cardiac surgery. Although one patient required an implantable defibrillator, all recovered without sequelae. One myocardial infarction in a patient receiving 35 IU/kg rFXIII-A(2) was identified by the Data Monitoring Committee after reviewing ECGs and cardiac enzymes. No other thromboembolic events were seen. Dosing with 25-50 IU/kg rFXIII-A(2) restored levels of FXIII to pre-operative levels, with a tendency towards an overshoot in receiving 50 IU/kg. rFXIII-A(2), in doses from 11.9 IU/kg up to 50 IU/kg, was well tolerated. For post-operative FXIII replenishment, 35 IU/kg of rFXIII-A(2) may be the most appropriate dose. PMID- 19806265 TI - Utility of 96-well plate aggregometry and measurement of thrombi adhesion to determine aspirin and clopidogrel effectiveness. AB - Aspirin and clopidogrel are key anti-thrombotic therapies. Results from platelet reactivity testing during therapy, have been shown to correlate with future events and would allow for the optimisation of therapy. However, there is little agreement among current tests and there remains a clear clinical need for a universal standardised test. It was the objective of this study to explore the potential of 96-well plate aggregometry as a definitive clinical test of platelet reactivity with respect to aspirin and clopidogrel. A small non-blinded trial of 16 healthy male volunteers assigned to seven days of aspirin (75mg/day) or clopidogrel (75mg/day) therapy. Blood was collected before and on day 7 of treatment. Platelet aggregation was measured using a 96-well plate based aggregation method, and thrombi adhesion measured by colourimetric assay. Platelet agonists used were ADP (0.1-30microM), arachidonic acid (0.03-1.3mM), collagen (0.1-30microg/ml), adrenaline (0.001-100microM), ristocetin (0.2 3mg/ml), TRAP6 amide (0.130microM) and U46619 (0.130microM). Concentration response curves were constructed to each agonist under the various conditions and used to extract data such as log EC(50), Hill slope, and area under the curve. These demonstrated low intra- and inter-assay variability and strong discrimination of drug effects. This study demonstrates the ability of the 96 well plate based aggregation and adhesion method to detect and differentiate between stable aspirin and clopidogrel treatment in healthy volunteers. Moreover, this assay marries the ability to test subjects or patients using a range of platelet agonists with more rapidity and ease than the current gold standard platelet assay, traditional light transmission aggregometry, making it a serious alternative assay for use in clinical settings. PMID- 19806266 TI - Is perioperative point-of-care prothrombin time testing accurate compared to the standard laboratory test? AB - There is no accepted way of measuring prothrombin time without time loss for patients undergoing major surgery who are at risk of intraoperative dilution and consumption coagulopathy due to bleeding and volume replacement with crystalloids or colloids. Decisions to transfuse fresh frozen plasma and procoagulatory drugs have to rely on clinical judgment in these situations. Point-of-care devices are considerably faster than the standard laboratory methods. In this study we assessed the accuracy of a Point-of-care (PoC) device measuring prothrombin time compared to the standard laboratory method. Patients undergoing major surgery and intensive care unit patients were included. PoC prothrombin time was measured by CoaguChek XS Plus (Roche Diagnostics, Switzerland). PoC and reference tests were performed independently and interpreted under blinded conditions. Using a cut-off prothrombin time of 50%, we calculated diagnostic accuracy measures, plotted a receiver operating characteristic (ROC) curve and tested for equivalence between the two methods. PoC sensitivity and specificity were 95% (95% CI 77%, 100%) and 95% (95% CI 91%, 98%) respectively. The negative likelihood ratio was 0.05 (95% CI 0.01, 0.32). The positive likelihood ratio was 19.57 (95% CI 10.62, 36.06). The area under the ROC curve was 0.988. Equivalence between the two methods was confirmed. CoaguChek XS Plus is a rapid and highly accurate test compared with the reference test. These findings suggest that PoC testing will be useful for monitoring intraoperative prothrombin time when coagulopathy is suspected. It could lead to a more rational use of expensive and limited blood bank resources. PMID- 19806267 TI - Inherited factor V deficiency associated with a novel heterozygous missense mutation (p.G493R) in a patient with excessive surgical bleeding. PMID- 19806268 TI - Fibrin gel structure obtained with a FVIIa analogue with enhanced FX-activating potential in haemophilia. PMID- 19806269 TI - Normal filopodia extension in VASP-deficient platelets upon activation by adhesive matrices or soluble agonists. PMID- 19806270 TI - Inferior vena cava agenesis: Association with bilateral lower-limb deep vein thrombosis in young males. PMID- 19806271 TI - Occupational exposure and incidence of respiratory disorders in a general population. AB - OBJECTIVE: To study the effect of occupational exposure on the incidence of adult asthma and six respiratory symptoms using a job exposure matrix (JEM). METHODS: From 1985, we conducted an 11-year community cohort study on the incidence of asthma and respiratory symptoms in Western Norway (N=2401, aged 15-70 years at baseline). The analyses in the current study were based on subjects < or =65 years at baseline, since those >65 years were unlikely to experience significant occupational exposures within the follow-up period. RESULTS: More women than men were exposed to biological dust (38% versus 29%), while more men were exposed to mineral dust (48% versus 19%), and gas or fumes (58% versus 53%). After adjusting for age, educational level, smoking, and previous occupational exposures, we found that high exposure to biological dust exposure was significantly related to a higher incidence of chronic and morning cough in men; for women, low exposure was related to attacks of dyspnea. Low exposure to workplace gas or fumes was significantly related to incident phlegm cough and attacks of dyspnea for women, while for men, high exposure was related to dyspnea grade 2. For the incidence of asthma and phlegm cough, after adjusting for all confounders, we found a significant interaction between workplace exposures and gender, where women had a higher risk of disease. CONCLUSIONS: Assessed by a JEM, occupational airborne exposure was weakly related to the incidence of asthma and respiratory symptoms, significantly more so for women than for men. PMID- 19806272 TI - Author's reply to Coggon commentary on epidemiological investigation of prognosis. PMID- 19806273 TI - Cancer incidence among large cohort of female Danish registered nurses. AB - BACKGROUND: Nurses are potentially exposed to carcinogens in their working environment. We investigated the risks for 21 types of cancers in Danish nurses. METHODS: We identified 92 140 female nurses from the computerized files of the Danish Nurses' Association. By record linkage, we reconstructed information on employment since 1964 using data from a national pension fund; information on vital status and reproduction was obtained from the Central Population Register. Each woman was followed-up from 1980-2003 in the Danish Cancer Registry. We calculated standardized incidence ratios (SIR) and 95% confidence intervals (95% CI). Using Poisson regression models, we made internal comparisons in subgroups of nurses, adjusting for potential confounders. RESULTS: We documented 8410 cancers during follow-up and found significantly increased SIR for breast cancer (SIR 1.1, 95% CI 1.1-1.2), cancers of the brain and nervous system (SIR 1.2, 95% CI 1.1-1.3), melanoma (SIR 1.2, 95% CI 1.1-1.3), and other skin cancers (SIR 1.2, 95% CI 1.1-1.2). Significantly decreased risks were observed for alcohol- and tobacco-related cancers. Nurses who were accredited by the Association after 1981 had significantly increased risks for thyroid cancer (SIR 1.9, 95% CI 1.3-2.5) and cancers of the brain and nervous system (SIR 1.5, 95% CI 1.2-1.9). Former nurses had significantly increased SIR for all cancers combined and breast cancer the first ten years after leaving the profession. In a Poisson regression analysis of breast cancer and duration of employment in hospitals, adjusted for reproductive factors, nurses had an increase risk the first 25 years of employment, but not for longer periods. CONCLUSION: The increased risk of breast cancer and the decreased risk of alcohol- and tobacco-related cancers support the findings of most other studies on nurses. The elevated risks for cancers of the breast, brain, nervous system, and thyroid warrant further study. PMID- 19806274 TI - Quality of working life and organizational performance--two sides of the same coin? AB - OBJECTIVE: This discussion paper presents the debate on the theoretical and practical claim that a simultaneous improvement in the quality of working life and organizational performance can be achieved by workplace development. METHODS: We discuss theories that support this claim or could provide additional support, and mention six "programs" on workplace development or social innovation in European countries. We present the results of evaluation studies in Finland, Germany, Ireland, and the Netherlands. RESULTS: A correlation of 0.50 was found between the quality of working life and organizational performance. All evaluations showed that the simultaneous improvement in the quality of working life and performance was achieved in a number of, but not all, projects. The percentage of successful projects is not always clear and depends on which aspects of quality of working life and performance were taken into account. Our estimation would be 25-50%. Which factors are important for success? With the exception of the commitment of top management and employee participation, other determinants were difficult to distinguish due to differences in research designs and programs. CONCLUSIONS: Important issues for discussion remain with respect to theory, research and practical approaches. Research should be more concerned with patterns of independent variables and how organizations deal with dilemmas. The most important pitfall appears to be taking a top-down as opposed to participatory approach involving employees and their supervisors in projects. National programs seem to be more effective than the initiatives of separate stakeholders groups. PMID- 19806275 TI - A systematic review of occupational safety and health business cases. AB - OBJECTIVE: Business cases are commonly developed as means to rationalize investment. We systematically reviewed 26 reported cases on occupational safety and health (OSH) interventions to assess if health and productivity arguments make a good business case. METHODS: To be included in the review, studies had to analyze the costs and benefits, including productivity, of an OSH intervention at the enterprise level. We searched Medline and Embase for studies and used Google search in addition. Two reviewers independently selected studies and extracted data. The intervention profitability was calculated in euros (euro in 2008) as the first year's benefits minus the total intervention costs per worker. The payback period was calculated as the intervention costs divided by the first year's benefits. RESULTS: We found three ex-ante and 23 ex-post cases. In 20 cases, the study design was a before-after comparison without a control group. Generally a 100% reduction of injuries or sickness absence was assumed. In two cases, productivity and quality increases were very large. The main benefit was avoided sick leave. Depreciation or discounting was applied only in a minority of cases. The intervention profitability was negative in seven studies, up to euro 500 per employee in 12 studies and more than euro 500 per employee in seven studies. The payback period was less than half a year for 19 studies. Only a few studies included sensitivity analyses. CONCLUSIONS: Few ex-ante business cases for management decisions on OSH are reported. Guidelines for reporting and evaluation are needed. Business cases need more sound assumptions on the effectiveness of interventions and should incorporate greater uncertainty into their design. Ex-post evaluation should be based preferably on study designs that control for trends at a time different from that of the intervention. PMID- 19806276 TI - The economic dimension of occupational health and safety. PMID- 19806277 TI - Socioeconomic position and low-back pain--the role of biomechanical strains and psychosocial work factors in the GAZEL cohort. AB - OBJECTIVE: To analyze the role that biomechanical strains and psychosocial work factors play in occupational class disparities in low-back pain in the GAZEL cohort. METHODS: Recruited in 1989, the GAZEL cohort members were employees of the French national company in charge of energy who volunteered to enroll in an annual follow-up survey. The study population comprised 1487 men who completed questionnaires in 1996 (past occupational exposure to manual material handling, bending/twisting, and driving), 1997 (psychosocial work factors), and 2001 (low back pain using a French version of the Nordic questionnaire for the assessment of low-back pain). Associations between low-back pain for >30 days in the preceding 12 months and social position at baseline (four categories) were described with a Cox model to determine prevalence ratios for each category. We compared adjusted and unadjusted ratios to quantify the contribution of occupational exposures. RESULTS: The prevalence of low-back pain for >30 days was 13.6%. The prevalence of low-back pain adjusted for age was significantly higher for blue-collar workers and clerks than for managers. The number of socioeconomic disparities observed was significantly reduced when biomechanical strains were taken into account; adjusting for psychosocial factors had little impact. CONCLUSION: In this population, occupational exposures--especially biomechanical strains--played an important role in occupational class disparities for persistent or recurrent low-back pain. PMID- 19806278 TI - Occupational asthma caused by inhalation of surfactant composed of amines. AB - OBJECTIVE: Occupational asthma (OA) is highly prevalent in industrialized countries and nearly 400 causal agents of this condition have been described to date. This study aims to describe the case of a patient who developed OA secondary to exposure to a surfactant agent comprised of alkylamine ethoxylate and a mixture of alkyleneoxy and ethylenediamine. METHODS: We present the case of a male worker in the meat industry suffering from OA resulting from exposure to a surfactant agent used to clean the animal carcass before it is quartered. We performed various tests on the individual, including: a chest computed tomography; total serum immunoglobulin E (IgE) and specific IgE tests against common pneumoallergens; pulmonary function studies; a methacholine test; and a specific inhalation challenge to the surfactant agent. RESULTS: The tests confirmed the diagnosis of OA. CONCLUSIONS: We discuss whether the amines present in the surfactant or the agent itself might be the cause of the condition. Because of the extensive use of surfactants in several types of industries, it is reasonable to think that their possible relationship with OA may have relevant health implications. PMID- 19806279 TI - Beliefs about back pain predict the recovery rate over 52 consecutive weeks. AB - OBJECTIVES: This study examined the course of low-back pain over 52 weeks following current pain at baseline. Initial beliefs about the inevitability of the pain's negative consequences and fear avoidance beliefs were examined as potential risk factors for persistent low-back pain. METHODS: On a weekly basis over a period of one year, 264 participants reported both the intensity and frequency of their low-back pain and the degree to which it impaired their work performance. In a multilevel regression analysis, predictor variables included initial low-back pain intensity, age, gender, body mass index, anxiety/depression, participation in sport, heavy workload, time (1-52 weeks), and scores on the "back beliefs" and "fear-avoidance beliefs" questionnaires. RESULTS: The group mean values for both the intensity and frequency of weekly low back pain, and the impairment of work performance due to such pain showed a recovery within the first 12 weeks. In a multilevel regression of 9497 weekly measurements, greater weekly low-back pain and impairment were predicted by higher levels of work-related fear avoidance beliefs. A significant interaction between time and the scores on both the work-related fear-avoidance and back beliefs questionnaires indicated faster recovery and pain relief over time in those who reported less fear-avoidance and fewer negative beliefs. CONCLUSIONS: Negative beliefs about the inevitability of adverse consequences of low-back pain and work-related, fear-avoidance beliefs are independent risk factors for poor recovery from low-back pain. PMID- 19806280 TI - The occupational safety and health scorecard--a business case example for strategic management. AB - OBJECTIVE: Human resources and health issues are crucial in terms of corporate competitiveness. However, systematic, continuous and strategically aligned occupational safety and health (OSH) management is scarcely applied in companies. One major reason for this could be the lack of generally accepted and standardised OSH control methods. Our objective was thus to conceptualize a method by which qualitative factors such as human resources and OSH aspects contribute to the performance or value-added layer of an organization. METHODS: We developed a business case based on the well-known and accepted Balanced Scorecard approach, which we adapted and applied to the management of OSH issues. The concept was implemented in the course of a comprehensive case study at a German automobile manufacturer. We gathered health as well as finance data in order to test which health-related indicators had an impact on financial performance. The demonstration of, and reporting on, how the promotion of workplace health contributes strategically to the organization is crucial for both health and human resource managers. RESULTS: Based on multivariate regression analyses, our main finding was that the Balanced Scorecard approach is an adequate means to control OSH issues in terms of strategic health management. Our analyses demonstrated that health-related interventions contribute significantly to performance aspects such as quality, productivity, absenteeism, and cost reduction. Therefore, the financial impact of health-related aspects / interventions could be demonstrated by means of the OSH scorecard. CONCLUSIONS: The availability and quality of health data within the context of overall corporate performance data needs to be improved in order to bridge OSH-related and performance issues of an organization. PMID- 19806281 TI - Comparative study of automotive, aircraft and biogenic emissions of aldehydes and aromatic compounds. AB - Air samples were collected in three well characterized locations in the city of Rio de Janeiro, Brazil: downtown, the idle and taxi way areas of the national airport and an urban forest, where the main emissions are from vehicular, aircraft and biogenic sources, respectively. Aldehydes and BTEX concentrations show a characteristic profile which may be attributed to the emission sources. Formaldehyde/acetaldehyde ratios, in the early morning, were 1.39, 0.62 and 2.22 in downtown, airport and forest, respectively. Toluene/benzene ratios, for downtown, airport and forest areas, were 1.11, 1.82 and 1.06, respectively. The results show that the impact of the urban emissions on the forest is negligible as well as the impact of aircraft emissions over the urban area. PMID- 19806282 TI - Hypothyroidism induced by polychlorinated biphenyls and up-regulation of transthyretin. AB - Polychlorinated biphenyls are environmental pollutants that are toxic to many biological systems. This study examined whether or not PCB126 and PCB114 have adverse effects on the serum thyroxine level and the serum proteome in rats. The results showed a lower serum total thyroxine level in the PCB126 and PCB114 treated groups than the control. Western blotting showed that the levels of transthyretin expression were significantly higher in the PCB-treated group than the control group. These results suggest that the PCB-mediated hypothyroidism is caused by the displacement of thyroxine from transthyretin. PMID- 19806283 TI - Comparison of the ability of organic acids and EDTA to enhance the phytoextraction of metals from a multi-metal contaminated soil. AB - Chelates have been shown to enhance the phytoextraction of metal from contaminated soil. In this study, we evaluated the ability of chelates to enhance the phytoextraction of metals by barnyard grass (Echinochloa crus-galli) from soils contaminated with multiple metals. The results revealed that EDTA increased the ability of barnyard grass to take up Cd, Cu and Pb, but that it resulted in increased soil leaching. Conversely, citric acid induced the removal of Cd, Cu and Pb from soil without increasing the risk of leaching. Furthermore, E.crus galli showed no signs of phytotoxicity in response to treatment with citric acid, whereas its shoot growth decreased in response to treatment with EDTA (p < 0.05). Taken together, these results demonstrate that citric acid is a good agent for the enhancement of the phytoextraction of metals. PMID- 19806284 TI - Hip protectors: recommendations for conducting clinical trials--an international consensus statement (part II). AB - INTRODUCTION: While hip protectors are effective in some clinical trials, many, including all in community settings, have been unable to demonstrate effectiveness. This is due partly to differences in the design and analysis. The aim of this report is to develop recommendations for subsequent clinical research. METHODS: In November of 2007, the International Hip Protector Research Group met to address barriers to the clinical effectiveness of hip protectors. This paper represents a consensus statement from the group on recommended methods for conducting future clinical trials of hip protectors. RESULTS AND CONCLUSIONS: Consensus recommendations include the following: the use of a hip protector that has undergone adequate biomechanical testing, the use of sham hip protectors, the conduct of clinical trials in populations with annual hip fracture incidence of at least 3%, a run-in period with demonstration of adequate adherence, surveillance of falls and adherence, and the inclusion of economic analyses. Larger and more costly clinical trials are required to definitively investigate effectiveness of hip protectors. PMID- 19806285 TI - Osteoporosis and venous thromboembolism: a retrospective cohort study in the UK General Practice Research Database. AB - In a retrospective cohort study using the General Practice Research Database (GPRD), there was a greater association of venous thromboembolism (VTE) in osteoporotic than in non-osteoporotic female patients. No greater association was shown in treated patients with strontium ranelate or alendronate compared to untreated osteoporotic female patients. INTRODUCTION: We explored the risk of VTE in usual practice in osteoporotic and non-osteoporotic women with and without anti-osteoporotic treatment. METHODS: A retrospective study was conducted using the GPRD in the UK. The cohorts consisted of untreated osteoporotic women (N = 11,546), osteoporotic women treated with alendronate (N = 20,084), or strontium ranelate (N = 2,408), and a sample of non-osteoporotic women (N = 115,009). Cohorts were compared using a Cox proportional hazards regression model. RESULTS: There was a significantly increased relative risk for VTE in untreated osteoporotic women versus non-osteoporotic women (annual incidence 5.6 and 3.2 per 1,000 patient-years, respectively; relative risk 1.75 [95% confidence interval (CI), 1.09-1.84]). Results were confirmed using adjusted models. The annual incidences of VTE in osteoporotic patients treated with strontium ranelate and alendronate were 7.0 and 7.2 per 1,000 patient-years, respectively, with no significant difference between untreated and treated patients whatever the treatment. Adjusted hazard ratios for treated versus untreated osteoporotic women were 1.09 (95% CI, 0.60-2.01) for strontium ranelate and 0.92 (95% CI, 0.63-1.33) for alendronate. CONCLUSION: This study shows a greater association of VTE in osteoporotic compared to non-osteoporotic patients, but does not show any greater association in treated patients with strontium ranelate or alendronate compared to untreated osteoporotic patients. PMID- 19806286 TI - Hip protectors: recommendations for biomechanical testing--an international consensus statement (part I). AB - INTRODUCTION: Hip protectors represent a promising strategy for preventing fall related hip fractures. However, clinical trials have yielded conflicting results due, in part, to lack of agreement on techniques for measuring and optimizing the biomechanical performance of hip protectors as a prerequisite to clinical trials. METHODS: In November 2007, the International Hip Protector Research Group met in Copenhagen to address barriers to the clinical effectiveness of hip protectors. This paper represents an evidence-based consensus statement from the group on recommended methods for evaluating the biomechanical performance of hip protectors. RESULTS AND CONCLUSIONS: The primary outcome of testing should be the percent reduction (compared with the unpadded condition) in peak value of the axial compressive force applied to the femoral neck during a simulated fall on the greater trochanter. To provide reasonable results, the test system should accurately simulate the pelvic anatomy, and the impact velocity (3.4 m/s), pelvic stiffness (acceptable range: 39-55 kN/m), and effective mass of the body (acceptable range: 22-33 kg) during impact. Given the current lack of clear evidence regarding the clinical efficacy of specific hip protectors, the primary value of biomechanical testing at present is to compare the protective value of different products, as opposed to rejecting or accepting specific devices for market use. PMID- 19806287 TI - Diagnosis and referral of rheumatoid arthritis by primary care physician: results of a pilot study on the city of Pisa, Italy. AB - The aims of the present study were to evaluate, in the city of Pisa: (1) the prevalence of rheumatoid arthritis; (2) the reliability of the prevalence estimated by primary care physicians, using the rheumatologist's diagnosis as the "gold standard" and (3) the economic impact of the disease. The Tuscany registry of primary care physicians constituted the framework from which a sample of subjects was selected. The rheumatoid arthritis (RA) subjects >18 years followed by each primary care physician constituted the population studied. Each general practitioner (GP) was asked to fill out a questionnaire regarding their patients affected by RA and to send it to the tertiary rheumatologic centre, where the diagnosis was confirmed/discarded, the clinical and epidemiological data were collected in a standardized form and a number of data for the estimation of costs were gathered. The estimated prevalence of RA was 5.1 per thousand (CI, 4.4-5.7). The reliability of general practitioners in the diagnosis of rheumatoid arthritis was on the whole 69%. However, when an analysis of every physician was carried out, a high degree of heterogeneity in the prevalence of RA per physician was found. Overall, the mean annual cost per patient with RA was estimated at about 5,878 euro (euro; median, 6,434 euro; inter quartile range, 669-7,052 euro), with a high variability mainly dependent on the degree of patient disability. More than 90% of the overall annual cost per patient was due to the medical and non medical direct components of costs. The prevalence of RA in Tuscany seems highly comparable with similar prevalence studies in Italy. The annual cost per patient with RA was highly variable and strictly dependent on the level of disability. More than 90% of the overall cost was due to the direct burden of costs. PMID- 19806288 TI - Anatomical basis of the risk of radial nerve injury related to the technique of external fixation applied to the distal humerus. AB - PURPOSE: Stabilization of humeral shaft and elbow fractures can be achieved with an external-fixator. Reports about nerve injuries associated with this procedure are rare in literature. Purpose of this anatomical study was to examine the relation of the radial nerve to distal humeral half pins. METHODS: Percutaneous insertion of external-fixator half pins was performed in 20 upper limbs of 20 cadavers, according to established technique, laterally in the distal humerus. RESULTS: Dissection of the upper limbs showed radial nerve injury in four of the 40 placed half pins. The proximal half pin impaled the nerve in one case and the distal half pin in three cases. Moreover the nerve was directly in contact with the pins in nine cases (five proximally, four distally). CONCLUSIONS: Insertion of external-fixator half pins in the distal humerus can easily injure the radial nerve. Thus we advocate a larger skin incision, blunt dissection to the lateral cortex of the humerus and retraction of soft tissue during half pin insertion. PMID- 19806289 TI - Shoulder bony landmarks location using the EOS low-dose stereoradiography system: a reproducibility study. AB - PURPOSE: To investigate the reproducibility of shoulder bony landmarks location using the EOS low-dose stereoradiography system, in order to validate this new tool for the study of gleno-humeral pseudo-kinematics. METHODS: An inter and intraobserver reproducibility study of shoulder bony landmarks location concerning 22 healthy volunteers. This study concerned the neutral position, arm at rest. Humerus and scapula were modeled with simple geometric shapes using specific software. Those shapes were positioned on A-P and lateral x-rays views. Images analysis of the 22 subjects was carried out three times (n(r) = 3), by two observers (n(o) = 2), for a total of n(tot) = 132 analyses. RESULTS: We obtained a very good reproducibility for the humeral head center and the diaphysis axis with 95% confidence interval (IC95%) inferior to 1.09 mm and 0.41 degrees, respectively. The uncertainty was higher for the lateral and medial epicondyles. Regarding the scapular bony landmarks, we observed a good reproducibility for the tip of the coracoid process, the inferior glenoid rim, and the axillar border with a 95% confidence interval lower than 2.13, 2.91 mm, and 3.67 degrees, respectively. The uncertainty was higher for the most postero-lateral point of the acromion and the superior glenoid rim. CONCLUSION: Our analysis of the x-rays obtained with the EOS low-dose stereoradiography system assessed the location reliability and reproducibility of specific scapular and humeral bony landmarks. This work opens the way to gleno-humeral pseudo-kinematics analysis using EOS imaging system. PMID- 19806290 TI - Health-related quality of life of elderly living in the rural community and homes for the elderly in a district of India. Application of the short form 36 (SF-36) health survey questionnaire. AB - The present investigation aimed to assess the health-related QoL (HRQoL) of elderly people living in two settings: (i) rural community and (ii) homes for the elderly in a district of South India. The data are drawn from elderly (>60 years of age) sampled from both settings. The short form 36-item health survey (SF-36) was administered to all respondents. The average scores for several domains, including total physical health, total mental health and overall health (total SF 36 score) were around 50, which can be interpreted as a moderate level of health related QoL. Residents living in a home for the elderly scored better in all domains except for role-physical and role-emotional. Though univariate analysis revealed some associations between characteristics of elderly SF-36 scores, the multiple regression analysis indicated that working status yields a significant but negative coefficient for total SF-36 score among community dwelling elderly. The elderly report that their lives are better when they are staying in homes for the elderly. Hence, despite the socio-economic conditions, provision of a better and conducive environment by setting up more charity-based homes for the elderly may be one of the options for relative betterment of the QoL of the elderly, particularly those who are socially and economically deprived. Finally, the study warrants the need of normative values of SF-36 for various population groups in India. PMID- 19806291 TI - [Underweight in elderly persons. A retrospective analysis of 3821 forensic autopsies in Hamburg]. AB - AIM AND METHOD: Based on the systematic analyses of a 10-year period at the Institute of Forensic Medicine at the University Clinic Hamburg-Eppendorf, the frequency of underweight in elderly persons at their time of death was retrospectively calculated. Body mass index (BMI) values <20 kg/m(2) were taken as indicative of underweight and the data of 1,551 women and 2,270 men, mean age 69 years (51-101 years) were analyzed. RESULTS: The prevalence of underweight was 15.4%. Low BMI values were more frequent in women than men (18.8 vs. 13.1%) and more frequent in persons needing professional nursing care at the time of death (28.9%). Underweight increased for the age range 70-79 from about 15% to over 38% for those 90 years and older at the time of death. Pressure sores were documented in 1.8% of cases. Persons in need of professional nursing care more often had pressure sores at their time of death. Furthermore, pressure sores were observed more often in underweight elderly in need of professional nursing care. CONCLUSION: Underweight at the time of death is observed more often in the very old. This is associated with the need for professional nursing care and also the occurrence of pressure sores. Less than 5% of persons with tumors were included. The results underscore the need of early detection of people at risk of malnutrition when growing older. PMID- 19806292 TI - [The meaning of work during early retirement]. AB - Since the mid-1990s, many German companies have used the part-time retirement scheme (Altersteilzeit) as an opportunity to release older workers. This was often consistent with the interests of the workers themselves. This paper explores whether workers who participate in this early retirement scheme would like to continue to work. A survey of all the workers in one company who participated in the part-time retirement scheme revealed that those already retired would more often like to continue working than employees who are still awaiting retirement. To a greater extent, early retirees would also like to return to work with shorter working hours or for a time-limited project. Results are explained with the affective forecasting bias that is based on the psychological decision theory and economic utility theory - the future utility of an action cannot be anticipated. The "reflective thinking" approach moreover invites the conclusion that the diverging interest of the two groups in continued employment may be due to the different stage of practical knowledge about the situation in (pre-)retirement. PMID- 19806293 TI - Exercise program for older patients with insulin-treated type 2 diabetes: long term effects on metabolic control and BMI. AB - BACKGROUND: Exercise is an important treatment for diabetes. In our study, we evaluated the acceptance of an exercise program and investigated the long-term effects of exercise on metabolic control and body mass index in older, insulin treated type 2-diabetic patients. PATIENTS AND METHODS: A total of 72 type 2 diabetic patients, aged >or=50 years, participated in an exercise program. At the beginning of the study and after 3 and 12 months, patients completed a questionnaire concerning current activity. Finally, the patients were divided into two groups: those with and without regular exercise. Metabolic control was compared in both groups. RESULTS: Even during the first workout, blood glucose levels fell. No patient suffered from severe hypoglycemia. Motivation for regular exercise increased from 33 to 77%, decreasing slightly to 65% at 12 months. The mean HbA 1c levels were similar in both groups. However, in the exercise group no weight gain was observed in contrast to 2.5% weight gain in the other group. Moreover, insulin requirements were considerably lower in the exercise group. CONCLUSION: The acceptance of regular exercise is also high in older patients with newly insulin-treated type 2 diabetes. Metabolic control was significantly improved in patients with and without regular exercise; however, in the exercise group, body weight did not increase and insulin requirements were significantly lower. PMID- 19806294 TI - Surgical treatment of hilar cholangiocarcinoma in the "new era": the Nagoya University experience. AB - OBJECTIVE: To review our surgical experience with hilar cholangiocarcinoma in the "new era." METHODS: The medical records of 428 patients with hilar cholangiocarcinoma who underwent treatment between 2001 and 2008 at the First Department of Surgery, the Nagoya University Hospital, were retrospectively reviewed. RESULTS: Of the 428 patients, 298 (70%) underwent surgical resection (R0, n = 220; R1, n = 70; R2, n = 8). Portal vein resection was performed in 111 (37%) patients, and hepatic artery resection was performed in 53 (18%) patients. Several different types of postoperative complications occurred in a total of 129 (43%) patients and led to 6 (2%) deaths. Of the 298 resected tumors, 206 (69%) were extrahepatic type tumors, and the remaining 92 (31%) were intrahepatic type tumors. Using the Bismuth classification criteria, we identified 15 (5%) type I, 21 (7%) type II, 120 (40%) type III, and 142 (48%) type IV tumors. The overall 1 , 3-, and 5-year survival rates for all 298 patients were 77, 49, and 42%, respectively. The survival rates were highest among the 197 patients with pM0 disease who underwent R0 resection. Patients in this subgroup had a 5-year survival rate of 52%. The 5-year overall survival rate for the 55 patients with pM0 disease who underwent R1 resection was 32%. The survival rate for patients who had pM1 disease and/or underwent R2 resection was the worst of all the subgroups, but was nonetheless significantly better than that of patients with unresectable tumors. The survival rate for patients who underwent vascular resection and reconstruction was unexpectedly better, with 5-year survival of >20%. CONCLUSIONS: The surgical approach to hilar cholangiocarcinoma has become more challenging in the new era. Nevertheless, surgical outcomes have been improved, with decreased morbidity and mortality rates being observed. Long-term survival has also steadily improved. These findings indicate that biliary surgeons should use an aggressive surgical strategy to treat this intractable disease. PMID- 19806295 TI - Hilar cholangiocarcinoma: the Memorial Sloan-Kettering Cancer Center experience. AB - BACKGROUND/PURPOSE: Hilar cholangiocarcinoma (HCCA) is a rare cancer with a low resectability rate, frequent recurrence after resection and an overall poor outcome. It is widely accepted that en bloc partial hepatectomy is a necessary part of the surgical therapy, but controversy surrounds other areas, including extent of lymphadenectomy and preoperative use of biliary drainage of the future liver remnant (FLR). This study analyzes the authors' experience with HCCA, emphasizing outcome after resection in a more recent cohort. METHODS: All patients with HCCA evaluated at Memorial Sloan-Kettering Cancer Center (MSKCC) since 1991 were included in the initial analysis. Outcome after resection was specifically assessed in patients submitted to operation between January 2001 and September 2008. Patient demographics, preoperative evaluation, resection type, margin status, lymph node status, complications, morbidity and survival were examined. Preoperative disease staging was performed in all patients according to the Blumgart classification. Separate analyses were conducted to assess the impact of preoperative biliary drainage on the FLR and the optimal lymph node harvest. Outcomes for resected patients were analyzed by Fisher's exact test and log rank tests. RESULTS: Three hundred and fifty-two patients with HCCA were evaluated since 1991, of which 118 were seen between 2001 and 2008. During this latter period, 105 (89%) patients underwent exploration, and of the 60 patients that underwent resection with curative intent, 48 (80%) had R0 resections. There were 3 perioperative deaths (5%), and 22 (28%) patients had complications. Patients with an R0 resection had the highest disease-specific survival followed by those with R1 resection when compared to unresected patients. The median follow-up period was 18 months. Classification by the Blumgart preoperative staging system predicted resectability and the likelihood of R0 resection. The benefit of pre-operative biliary drainage of the FLR appeared to be limited to patients with a predicted FLR volume of <30%. In patients with node-negative tumors, survival was greater in those with more than 7 lymph nodes harvested. CONCLUSIONS: R0 resection including hepatectomy with negative lymph nodes is feasible in the majority of patients with resectable HCCA. This strategy is associated with a prolonged disease-specific survival. PMID- 19806296 TI - Carbon dioxide-enhanced virtual MDCT cholangiopancreatography. AB - BACKGROUND/PURPOSE: We investigated the feasibility of new carbon dioxide enhanced virtual multidetector computed tomography (MDCT) cholangiopancreatography (CMCP) for intraluminal exploration in 73 patients with hepatobiliary and pancreatic disease. METHODS: CMCP was performed via a percutaneous or transpapillary drainage tube, and, synchronously, intravenous contrast material was employed for virtual angiography; three-dimensional (3D) virtual reality was incorporated using OsiriX and Fovia applications. The capability of carbon dioxide to delineate the biliary and pancreatic system was evaluated. RESULTS: All CMCPs showed complete technical success; complications including pancreatitis or pain never occurred. The incidences of visible third- and fourth-order biliary branches were 100 and 86.0%. The capability of carbon dioxide to pass an obstruction through an occluded hilar bile duct malignancy was 80.0%; it provided feasible information on additional bile duct segments. The full extent of the gallbladder was depicted in 72.7% of the studies. Minimum 2-mm lesions of biliary stones or gallbladder polyps were clearly detected. The main or second-order pancreatic ducts were visible in 100 and 83.3% of the studies, respectively. Carbon dioxide enabled the replacement of mucin and pancreatic juice and facilitated the detection of cystic lesions of intraductal papillary mucinous neoplasm (IPMN) in 75.0% of the studies. We succeeded in achieving 3D spatial recognition of vascular structures in the cholangiopancreatic region, through the fusion of CMCP and 3DCT arteriography and venography in a single image scanning, and radiation time was decreased. This combined modality proved to be feasible for planning operations and for image-guided navigated surgery in the resection of a malignancy. CONCLUSIONS: To our knowledge, this is the first report to demonstrate the diagnostic accuracy of carbon dioxide MDCT cholangiopancreatography and the use of this modality for depicting biliary, pancreatic, and fusion blood vessels simultaneously. Carbon dioxide possesses many advantages over conventional iodinated contrast agents, and it might replace more invasive diagnostic measures in the near future. PMID- 19806297 TI - Integrated MR-laparoscopy system with respiratory synchronization for minimally invasive liver surgery. AB - PURPOSE: The laparoscope has been invaluable in minimally invasive surgery, but provides only a surface view of target tissue; therefore it is lacking internal tissue information. In combination with the laparoscope for visualizing the cross sectional view of the tissue, MRI is superior to ultrasonography or X-ray CT, because of its high soft-tissue contrast, arbitrary slice orientation and lack of radiation properties. Thus, we propose an integrated MR-laparoscopy system with a respiratory-synchronized navigation. METHODS: A transmit/receive RF coil for localized MR imaging with a 0.5 T open-MRI was mounted onto the tip of an MR compatible laparoscope. The signal detection of the coil was examined with an excised porcine liver sample, an agar phantom and the abdominal wall of a healthy volunteer. A real-time navigation system to compensate for respiratory motion was developed, and examined with a healthy volunteer. RESULTS: The SNRs of the local MR images were 112, 62, and 62 in the liver sample, phantom, and volunteer. The navigation system successfully displayed the scope view, scope location and orientation, and MR images with respiratory-synchronized real time operation. CONCLUSIONS: The MR-imaging and synchronization function of the proposed system seemed to be helpful for laparoscopic surgery. PMID- 19806298 TI - Endoscopic ultrasonography-guided biliary drainage. AB - BACKGROUND AND PURPOSE: Endoscopic ultrasonography-guided biliary drainage (EUS BD) has been developed as an alternative drainage method in patients with obstructive jaundice. EUS-BD is divided into EUS-guided choledochoduodenostomy (EUS-CDS), EUS-guided hepaticogastrostomy (EUS-HGS) and EUS-guided gallbladder drainage (EUS-GBD). The aim of this review is to focus on the current status and limitations of EUS-BD. METHODS: A systematic review was performed to evaluate EUS BD. MEDLINE, EMBASE and manual searches were performed to identify the pertinent English language full articles. RESULTS: The high success rate without fatal adverse events for EUS-CDS (93%; 28/30) and EUS-HGS (97%; 28/29) suggest the feasibility and safety of the procedures in high-volume endoscopic centers adopting various procedural techniques. Although the number of reported cases was very small, the success rate of EUS-GBD was high (100%; 14/14), without serious adverse events. CONCLUSIONS: Although all procedures require further assessment in a larger cohort of patients, including comparative studies between EUS-CDS or EUS-HGS versus PTBD, and EUS-GBD versus PTGBD, EUS-BD may be a promising procedure for the treatment of obstructive jaundice. However, dedicated devices for EUS-guided drainage are needed for reliable procedures. PMID- 19806299 TI - Intraoperative exploration of biliary anatomy using fluorescence imaging of indocyanine green in experimental and clinical cholecystectomies. AB - BACKGROUND/PURPOSE: We evaluated the usefulness of intraoperative exploration of the biliary anatomy using fluorescence imaging with indocyanine green (ICG) in experimental and clinical cholecystectomies. METHODS: The experimental study was done using two 40-kg pigs and the clinical study was done in 12 patients for whom cholecystectomy was planned from January 2009 to June 2009. Initially we used a laparoscopic approach for the evaluation of fluorescence imaging of the biliary system in the two pigs. Then the clinical study was started on the basis of these experimental results. ICG (1.0 ml/body of 2.5 mg/ml ICG) was infused 1-2 h before surgery. With the subjects under general anesthesia we observed in real time the condition of the biliary tract under the guidance of fluorescence imaging employing an infrared camera or a prototype laparoscope. ICG was added intravenously to observe the location or flow condition of the cystic artery. RESULTS: We obtained a clear view of the biliary tract and the location of the cystic duct in the two pigs. Local compression with a transparent hemispherical plastic device was effective for offering a clearer view. The biliary tract, except for the gallbladder, was clearly recognized in all clinical subjects. Local compression with a transparent hemispherical plastic device for open cholecystectomy and a flat plastic device for laparoscopy provided clearer visualization of the confluence between the cystic duct and common bile duct or common hepatic duct. The location of the cystic artery was revealed after division of the connective tissues, and the flow condition of the cystic artery was confirmed 7-10 s after intravenous re-infusion of ICG. There were no adverse events related to the intraoperative procedure or the ICG itself. CONCLUSIONS: This method is safe and easy for the identification of the biliary anatomy, without requiring cannulation into the cystic duct, X-ray equipment, or the use of radioactive materials. Although fluorescence imaging is still at an early stage of application in comparison with ordinary intraoperative cholangiography, we expect that this method will become routine, offering a lower degree of invasiveness that will help avoid bile duct injury. PMID- 19806300 TI - Recent advances in visualization, imaging, and navigation in hepatobiliary and pancreatic sciences. AB - BACKGROUND/PURPOSE: Recent introduction of multi-detector CT (MDCT) and high speed magnetic resonance (MR) imaging have dramatically advanced visualization and imaging technology in diagnostic and therapeutic strategy in hepatobiliary pancreatic disease. However, image diagnostics have progressed with a background of the essence of anatomy, pathology, and physiology. It is important to object the reflection of the patient's condition and pathology of each disease and remove pattern recognition in what they were depicted as an image. Visualization plays another important role in various medical diagnostics. Trends in scientific visualization will depend on advancements in molecular technology and computer hardware as well as trends in engineering disciplines. METHODS: In this special issue, the recent advances in visualization and imaging in the field of hepatobiliary and pancreatic sciences are featured including application of advanced visualization techniques, data management, data compression, feature extraction. RESULTS: We discuss the potential benefits of new technologies and procedures in hepatobiliary and pancreatic areas, that are circulating tumor cells, MR imaging for hepatocellular carcinoma, indocyanine green using fluorescence under infrared light observation, carbon dioxide enhanced MDCT virtual cholangiopancreatography, endoscopic ultrasonography-guided biliary drainage, natural orifice translumenal endoscopic surgery, MR-laparoscopy, and image overlay navigation surgery by OsiriX. CONCLUSION: Some of the recent trends are discussed in terms of visualization and imaging in hepatobiliary and pancreatic sciences. The goal in using visualization is to assist existing scientific procedures by providing new insight through visual representation. PMID- 19806301 TI - Laparoscopic and robotic donor pancreatectomy for living donor pancreas and pancreas-kidney transplantation. AB - Pancreas transplantation is a widely accepted procedure that can efficiently restore euglycemia and prevent progression of complications. In most instances, the limiting factor for deceased donor organ transplantation is the availability and quality of the available organs. Living donor pancreas transplant was introduced at the University of Minnesota in 1979. Because of the potential risks for the donor and the technical challenges in the recipient operation, this procedure has not become very popular since then. In 1999, in the attempt to decrease the morbidity associated with open distal pancreatectomy, the first laparoscopic donor distal pancreatectomy with hand-assisted technique was performed at the same institution. In 2000, the FDA approved the robotic surgical system Da Vinci for general use. Since then, the system has been extensively used at our institution to perform living donor nephrectomy. The only case reported worldwide of robotic distal pancreatectomy and nephrectomy for living donor pancreas-kidney transplantation was successfully performed by our team in 2006 at the University of Illinois at Chicago and proved as a promising technique. The application of minimally invasive techniques has allowed an increased acceptance of the procedure among potential donors and may, therefore, increase the number of donors for this life-saving transplant. The initial results are encouraging and clearly prove feasibility. PMID- 19806302 TI - Teriparatide is safe and effectively increases bone biomarkers in institutionalized individuals with osteoporosis. AB - Institutionalized adults with severe developmental disabilities have a high rate of minimal trauma and appendicular fracture. There is little information about osteoporosis treatment in this population. In this efficacy and safety study, men and women with severe developmental disabilities and osteoporosis received 20 mcg teriparatide subcutaneously daily for 18-24 months. Markers of bone formation [procollagen type 1 intact N-terminal propeptide (P1NP)] and resorption [C telopeptide (CTx)] were measured at three-month intervals. Serum calcium was measured at two-week intervals for 12 weeks and thereafter at three-month intervals. Twenty-seven individuals received at least one injection. The incidence of hypercalcemia was 11.1% but was persistent and led to medication discontinuation in only one participant. Biomarkers of bone formation increased rapidly, doubling by three months. At 12 months, P1NP and CTx remained elevated from baseline; P1NP had risen from 66.95 +/- 83.71 microg/l (mean +/- SD) to 142.42 +/- 113.85 microg/l (P = 0.05), and CTx had increased from 0.377 +/- 0.253 to 1.016 +/- 1.048 ng/ml (P = 0.01). The majority of participants had an increase in P1NP of over 10 microg/l. In conclusion, teriparatide is safe and effective in developmentally disabled institutionalized adults. Serial calcium measurements are warranted, particularly during the first three months of therapy. PMID- 19806303 TI - Outcome correlates with blood distribution in subarachnoid hemorrhage of unknown origin. AB - PURPOSE: Between 15 and 30 % of patients with subarachnoid hemorrhage (SAH) have no bleeding source and usually have a benign clinical course and outcome. The objectives of this study were to classify the pattern of blood distribution on initial computed tomography (CT) and to correlate it with clinical outcome in aneurysmal (ASAH) and SAH of unknown origin (SAHuO). METHODS: We reviewed 112 CTs of SAHuO and 104 CTs of ASAH patients. Blood distribution was classified according to a new grading system (type 0-4) and correlated to Hunt and Hess (H&H) grade and modified Rankin scores (MRS) at short- and long-term follow-up. RESULTS: Fifty percent of 112 SAHuO patients were classified as type 0 (no visible blood on CT) or 1 (blood restricted to prepontine cisterns). Most ASAH patients presented with bleeding into the lateral Sylvian fissure (66%; type 3) or with intracerebral hemorrhage (27%; type 4) whereas types 0 and 1 were not observed. SAHuO patients were in better clinical condition on admission than ASAH patients (p < 0.0001). H&H grades of SAHuO patients correlated with the amount of subarachnoid blood according to the new classification (p = 0.004). Short-term outcome was obtained from 100% and long-term outcome from 95% patients (follow-up 29 +/- 31 months). Short- and long-term MRS correlated with blood distribution in SAHuO patients (p = 0.012) and was significantly better than in ASAH patients (p < 0.0001). No correlation was observed between blood distribution, H&H grade, and short- and long-term outcome in aneurysmal patients. CONCLUSIONS: In SAH of unknown origin, a new classification allows to predict outcome based on the extent of blood on CT. PMID- 19806304 TI - Surgical exposure of the carotid artery for endovascular interventional procedures. AB - BACKGROUND: Transfemoral approach for endovascular interventions is not always possible in cases of unfavorable anatomy. We report our experience using a transcervical approach with carotid cut down and direct, controlled puncture of the carotid artery. METHODS: Four patients underwent surgical exposure of the carotid artery for endovascular procedures. One patient had retrograde placement of a stent in the common carotid artery, and three patients had coiling of an intracranial aneurysm. After the endovascular procedure, the sheath was removed and the vessel was closed, under direct visualization. RESULTS: The technique allowed access to extracranial and intracranial lesions. There were no access site complications. There were no access site-related cardiac, systemic, or neurologic events. CONCLUSIONS: Transcervical access with surgical exposure of the carotid artery for direct and controlled vascular puncture is an effective alternative for endovascular extracranial and intracranial procedures in patients in whom the femoral route cannot be used. PMID- 19806305 TI - Influences of stents on the outcome of coil embolized intracranial aneurysms: comparison between a stent-remodeled and non-remodeled treatment. AB - PURPOSE: The influences of a stent on the outcome of embolized aneurysms are not well known. The purpose of this study is to assess and compare the angiographic follow-up (FU) results of stent-remodeled (StR) and non-remodeled (nonR) coil embolization of intracranial aneurysms. METHODS: Thirty-seven StR patients with FU digital subtraction angiography (DSA) were matched with 37 nonR patients according to presentation (ruptured or unruptured), location of the aneurysm (sidewall or bifurcation), initial sac occlusion (complete/near complete or incomplete/failure), and the sac size (largest diameter/less than 2 mm difference). The baseline clinical and aneurysm characteristics, initial/FU treatment success rates, and FU DSA findings between the StR and nonR groups with regard to the morphologic features (size and location) of the aneurysms were assessed and compared. RESULTS: The StR group showed more frequent progressive thrombosis compared to the nonR group (56.8 vs. 27.0%; p < 0.05). These differences were more prominent for small aneurysms (70 vs. 25%; p < 0.05). Small aneurysms showed a lower rate of FU recanalization in the StR group compared to the nonR group (0 vs. 25%; p < 0.05). Sidewall aneurysms also showed a higher rate of progressive thrombosis in the StR group (61.1 vs. 27.8%; p < 0.05). CONCLUSIONS: Coil embolization is an effective treatment method for intracranial aneurysms, however, changes on follow-up may affect the long-term outcome. Stent remodeling may aid in preserving and enhancing the treatment durability especially in small aneurysms. PMID- 19806306 TI - Intracranial pressure pulse amplitude during changes in head elevation: a new parameter for determining optimum cerebral perfusion pressure? AB - OBJECTIVE: During short-term postural changes, the factors determining the amplitude of intracranial pulse pressure (ICPPA) remain constant, except for cerebrovascular resistance (CVR). Therefore, it may be possible to draw conclusions from the ICPPA onto the cerebrovascular resistance (CVR) and thus the relative change in cerebral perfusion pressure (CPP). METHODS: Age, sex, disease, Glasgow Coma Scale score, placement of ventricular drain, blood gas analysis, and parameters of airway management were prospectively recorded in 40 patients. The changes in intracranial pressure (ICP), CPP, mean arterial pressure (MAP), and ICPPA at head elevations of 0 degrees, 30 degrees, and 60 degrees were measured and analyzed online. Status of cerebrovascular autoregulation was checked using the pressure-reactivity index (PRx). RESULTS: Altogether 36 subjects fulfilled the study conditions. Three patients had positive PRx indicating disturbed autoregulation and were excluded. Thus, 33 were left for analysis (18 females and 15 males). All of them were sedated and mechanically ventilated with Glasgow Coma scores ranging from 3-8. During change in head elevation from 0 degrees to 60 degrees, we found a significant (p < 0.05) improvement of the ICP, an increase of the ICCPA, a reduction of the MAP, and a decrease in the CPP. Increasing ICPPA was linked to decreasing CPP (0 degrees to 60 degrees, r = -0.42, p < 0.05). CONCLUSIONS: Head elevation is an important part of the ICP and CPP therapy in neurointensive care. When searching for the patient-specific optimum upper body position, ICPPA may provide additional information. Providing that the cerebral autoregulation is intact, the lowest ICPPA of a patient corresponds to the individual upper body position with the highest CPP. PMID- 19806307 TI - Prediction of early secondary complications in patients with spontaneous subarachnoid hemorrhage based on accelerated sympathovagal ratios. AB - PURPOSE: The development of secondary complications following spontaneous subarachnoid hemorrhage (SAH) largely depends on sympathetic overexcitation. The roles of vagal activities, however, are poorly defined. Because both components of the autonomic nervous system can be explored in the frequency domain of heart rate variability (HRV), the present study aimed to determine the dynamic evolution of autonomic activities and to identify patients at high risk for complications following hemorrhage. METHODS: Thirty patients with SAH were enrolled in our study. Those who suffered from symptomatic vasospasm, cerebral infarction, neurogenic pulmonary edema, or early mortality within 1 week of ictus were categorized into the complication group. Spectral analysis of HRV explored three important indices of sympathetic and vagal modulations: low-frequency (LF), high-frequency (HF), and LF/HF ratios. Patterns of HRV dynamics within the first 3 days were compared between complication and non-complication groups. The group trends, estimated by the slopes of HRV changes, were determined for further univariate and multivariate analysis. RESULTS: Our study showed that daily HRV in the complication group exhibited an approximately 2.7-fold increase of sympathovagal ratio (denoted by LF/HF). This resulted from reciprocal changes of sympathoexcitation (LF) and vagal withdrawal (HF). Multivariate analysis revealed that LF/HF slope, an indicator of the trend of sympathovagal change, was an independent variable significantly associated with the development of complications. CONCLUSIONS: This study confirmed that during early SAH period, patients with and without complications presented different patterns of sympathovagal changes. LF/HF slope during the first 3 days was a significant predictor of secondary complications after SAH. PMID- 19806308 TI - Cerebellopontine angle arachnoid cyst containing ectopic choroid plexus--case report. AB - We present a rare and interesting case of a cerebellopontine angle cyst containing ectopic choroid plexus tissue in a 26 year-old female. Surgical resection was performed, and histological examination confirmed the presence of choroid plexus in the cyst wall. This is the first reported case of ectopic choroid plexus at the cerebellopontine angle in an adult. We present the case and review the literature. PMID- 19806309 TI - Assessment of the variability in the anatomical position and size of the subthalamic nucleus among patients with advanced Parkinson's disease using magnetic resonance imaging. AB - PURPOSE: Targeting of the subthalamic nucleus (STN) during deep brain stimulation (DBS) surgery using standard atlas coordinates is used in some centers. Such coordinates are accurate for only a subgroup of patients, and subgroup size depends on the extent of inter-individual variation in STN position/size and degree to which atlas represents average anatomical relations. Few studies have addressed this issue. METHODS: Sixty-two axial T(2)-weighted magnetic resonance (MR) images of the brain (1.5 T) were obtained before STN-DBS in 62 patients (37 males) with Parkinson's disease using a protocol optimized for STN visualization. Image distortion was within sub-millimeter range. Midcommissural point (MCP) derived coordinates of STN borders, STN center, and other brain landmarks were obtained using stereotactic software. MR-derived measurements were compared to Schaltenbrand and Wahren Atlas. RESULTS: We evaluated 117 best-visualized STNs. STN dimensions and coordinates of its center were highly variable. STN lateral coordinate ranged 8.7 mm-14.5 mm from MCP, A-P coordinate 3.5 mm posterior to 0.5 mm anterior to MCP, and vertical coordinate 1.3 mm-6 mm below MCP. The antero posterior nucleus dimension varied by 8 mm and lateral-medial dimension by 5.8 mm. Differences between mean values of MR-derived data sets and Atlas values were statistically significant but moderate, excluding AC-PC length, for which the Atlas value was below the 1st percentile of the MR data set. The STN lateral coordinate strongly correlated with the width of the third ventricle (r = 0.73, p < 0.001). CONCLUSIONS: It is now possible to directly evaluate STNs at 1.5 T with minimal image distortion, which reveals variation in STN position and dimensions in the range of nucleus size. This puts under question the rationale of use of standard STN coordinates during DBS surgery. PMID- 19806310 TI - Cerebral venous thrombosis with dural arteriovenous fistulas and antiphospholipid syndrome: a case report. PMID- 19806311 TI - Psychiatric symptoms in Parkinson's disease assessed with the SCL-90R self reported questionnaire. AB - The frequency of psychopathological symptoms in patients with Parkinson's disease (PD) is often underestimated because of the lack of comprehensive evaluation tools. A total of 486 consecutive non-demented PD patients completed the Symptom Checklist 90 Revised (SCL-90R) self-reported questionnaire, a validated tool for the assessment of psychopathological symptoms on nine dimensions. Somatization, depression, anxiety and obsessive-compulsive behaviors were reported by nearly half of the PD patients. They were more likely to occur in females. Disease related factors such as duration, severity and daily dosages, but not type of dopaminergic medications, were associated with the occurrence of these symptoms. Psychopathological features are frequent in PD and their occurrence is underlined by disease-related factors. PMID- 19806312 TI - Isolated ataxia after pure left insular cortex infarction. AB - Acute insular infarction, due to its anatomic and functional complexity and wide connections, may present with various clinical presentations, such as somatosensory deficits, gustatory disorder, vestibular-like syndrome, cardiovascular disturbances, neuropsychological disorders, movement disorders, autonomic dysfunction and empathy impairment. However, there was no mention of the symptoms involving the cerebellar system in the related literature. We present a case of pure left insular cortex infarction with isolated truncal ataxia and demonstrate a crucial relationship between the left insular cortex and the cerebellar system. The possible connections are through the spinocerebellar and dentatorubrothalamic pathway. In conclusions, left insular cortex lesions should be considered in the differential diagnosis of isolated truncal ataxia. PMID- 19806313 TI - Thoracic extradural haematoma after epidural anaesthesia. AB - The administration of analgesics to the thoracic spine is established practice in the operating room, minimizing the need for systemic anaesthetic administration during thoracic surgery. Complications arising from thoracic epidural anaesthesia are uncommon but potentially disastrous. Here, we report the case of a 43-year old woman who developed a thoracic epidural haematoma with paraplegia a few hours after the removal of an epidural catheter. The patient underwent emergency thoracic laminectomy and clot evacuation. After a 4 months period, there was almost complete neurological recovery. Epidural haematoma is a rare complication that must be heeded and urgently treated in case of clinical deterioration after the epidural analgesia. PMID- 19806314 TI - A case report of cranial polyneuropathy in Behcet's disease. AB - Behcet's disease (BD) is a multisystem relapsing inflammatory disorder. Nervous system involvement is the most serious manifestation of BD. Neuro-Behcet's disease (n-BD) is classified into parenchymal meningoencephalitis pattern and non parenchymal vascular patterns. Isolated meningitis is rarely the sole presenting feature of n-BD. We report a 19-year-old man with BD who presented with cranial polyneuropathy secondary to aseptic meningitis. He made a gradual though incomplete recovery with steroid therapy. Cranial polyneuropathy secondary to meningeal involvement can be a presentation of n-BD, and Behcet's disease should be a consideration in patients with idiopathic cranial polyneuropathy. PMID- 19806315 TI - Advanced age as a contraindication to microvascular decompression for drug resistant trigeminal neuralgia: evidence of prejudice? AB - Due to the alleged higher risk of complications of microvascular decompression for trigeminal neuralgia in elderly we evaluated its age-related results. A retrospective analysis of clinical outcome and complications was performed in 476 patients affected by drug-resistant trigeminal neuralgia who underwent microvascular decompression. As much as 117 patients older than 65 years (Group 1) and 359 under the age of 65 (Group 2) were followed-up for a period of 7-138 months. Pain relief was complete without medication in 84.5% in Group 1. Morbidity included slight trigeminal hypoesthesia in 5.8%; severe hearing loss in 0.9%; CSF leakage in 4.2%; transient diplopia in 3.4%; and posterior fossa subdural hematoma in 0.8% of these patients. Mortality was null. No statistically significant differences were observed between Groups 1 and 2. These findings seem to support the idea that microvascular decompression is not a dangerous surgical procedure in patients over the age of 65 years. PMID- 19806316 TI - Cerebellar degeneration and ocular myasthenia gravis in a patient with recurring ovarian carcinoma. AB - Myasthenia gravis (MG) and paraneoplastic cerebellar degeneration (PCD) are immune-mediated syndromes that can represent paraneoplastic disorders. We report a patient with history of ovarian carcinoma that presented with ptosis, diplopia and gait ataxia. Neurophysiological examination and laboratory tests revealed the presence of MG and PCD. An integrated FDG-PET/contrast-enhanced CT scan showed tumor recurrence. This is to the best of our knowledge the first association of MG and PCD with recurring ovarian carcinoma. PMID- 19806317 TI - An assessment of socio-economic inequalities in health among elderly in Greece, Italy and Spain. AB - OBJECTIVE: This study explores socio-economic inequalities in health among Mediterranean people aged 50 or higher. METHODS: The data used in the analysis come from the Survey of Health, Ageing and Retirement in Europe, wave 1, release 2; the sample includes 2,671 Greek, 2,502 Italian and 2,343 Spanish persons. Seven health indicators are examined using age-sex standardized prevalence rates and logistic regression models. Concentration indices are also computed for self rated health (SRH). RESULTS: Socio-economic position of individuals declines with age. Persons of lower socio-economic position experience worse health in all instances. Independently of education and gender, Greek persons display the lowest prevalence rates for SRH and physical and depressive symptoms, Spanish exhibit the highest rates for chronic conditions, and Italians perform better regarding functional limitations. Within-country analysis shows that the magnitude of socio-economic inequalities in SRH is greatest in Greece, followed by Spain and lastly by Italy. CONCLUSIONS: The analysis reconfirms the advantage of high over low socio-economic position for all countries and health indicators and proves education as an important correlate compared to wealth and income among the elderly. PMID- 19806318 TI - Procalcitonin as an early marker of bacterial infection in neutropenic febrile children with acute lymphoblastic leukemia. AB - OBJECTIVE AND DESIGN: The aim of this study was to assess the value of procalcitonin (PCT), C-reactive protein (CRP), tumor necrosis factor-alpha (TNF a), interleukin (IL)-1b, IL-8, and soluble TNF receptor II (sTNFRII) in early and rapid diagnosis of infection in neutropenic children with acute lymphoblastic leukemia (ALL) and to distinguish bacterial from viral infections. PATIENTS: The study included five groups (A, B, C, D, and E) of children with ALL undergoing intensive chemotherapy. Groups A and B consisted of neutropenic children with bacterial and viral infection, respectively. Groups C and D consisted of nonneutropenic children with bacterial and viral infection, respectively. Group E consisted of children without neutropenia and without fever. METHODS: In all groups, blood samples were collected upon admission and then for 7 days on a daily basis. Levels of CRP, PCT, TNF-a, IL-1b, IL-8, and sTNFRII were determined in all blood samples. RESULTS: We found a highly significant difference in PCT levels between bacterial and nonbacterial episodes. Sensitivity and specificity of PCT were 94 and 96.5%, respectively. CONCLUSIONS: Serial measurement of PCT levels on a daily basis seems to be helpful for early prediction of severe bacterial infections, monitoring febrile episodes regarding response to antibiotic therapy, and early detection of complications in the infectious process. PMID- 19806319 TI - Pituitary metastasis of Merkel cell carcinoma. AB - Merkel cell carcinoma (MCC) is a malignant neuroendocrine tumor of the skin that demonstrates a remarkable tendency to metastasize. However, only a few cases of MCC brain metastases have been reported in the literature. We here present a unique case of a pituitary metastasis of MCC in a 65-year-old patient with a history of pituitary adenoma. This case is particularly novel due to the fact that the primary site of the MCC is unknown. PMID- 19806320 TI - The cell growth inhibitory transcription factor C/EBPdelta is expressed in human meningiomas in association with low histological grade and proliferation index. AB - CCAAT/enhancer binding protein (C/EBP) delta is a transcription factor which has been demonstrated to mediate the growth arrest of mammary and prostate cancer cell lines. It is induced by several stimuli including inflammatory cytokines. In this study, C/EBPdelta immunohistochemical expression was assessed in 49 meningiomas of different histotype and grade and correlated with a variety of clinico-pathological data and with the overall and recurrence-free survival of the patients. Positive staining was observed in the nuclei of neoplastic cells in 22 out of the 49 cases analyzed. C/EBPdelta expression was significantly associated with a low histological grade and proliferation index, reflected by low Ki-67 labeling index (LI) and mitotic activity, and with the presence of intra-tumoral inflammatory infiltrate and the absence of necrosis. In addition, the absence of C/EBPdelta was significantly correlated with a shorter disease free interval. Our findings suggest that C/EBPdelta expression may prevent the development of recurrences by inhibition of neoplastic growth in meningiomas. If further studies confirm its induction by inflammatory mediators, this might be exploited in novel therapies to prevent recurrences in meningiomas. PMID- 19806321 TI - Clinical and epidemiological characteristics of pediatric gliosarcomas. AB - Gliosarcoma (GS) is a glioblastoma with a sarcomatous component that is presumed to be a metaplastic differentiation of glioma cells. We studied the clinical relevance of this histological glioblastoma subentity within the pediatric population. We obtained patient data from the German HIT-GBM database, which contains clinical data for more than 600 pediatric patients with centrally reviewed high-grade gliomas. By applying defined inclusion criteria (diagnosis of GS proven by central neuropathological review; patient age 0 to 21 years), four patients were identified. In addition, after a review of the English medical scientific literature, 19 additional cases were found. The relative frequency of GS in the German HIT-GBM database was only 1.9%. In the whole series of 23 pediatric GS patients, including previously reported cases, the male-to-female ratio was 1.2:1. GS was found in all pediatric age groups with a median age of 11 years, but there was an unexpectedly high accumulation in infants (6 of 23 <3 years of age, 26%). GS showed a strong predilection of the cerebral hemispheres (22 out of 23 cases). Increased intracranial pressure was the leading symptom of a short clinical history with a median duration of 0.7 month. Interestingly, six patients (26%) were reported with a history of cranial radiotherapy prior to GS diagnosis. In 60% of the GS patients in our series, gross total resection was achieved. Median overall (OS) and event-free survivals (EFS) of the total cohort were 12.1 and 9.8 months, respectively. In conclusion, GS is a very rare tumor entity in children. Literature review suggests a relatively higher incidence in infants and in patients with a previous history of radiotherapy. PMID- 19806322 TI - Identification of the role of Smad interacting protein 1 (SIP1) in glioma. AB - Glioma is an extremely aggressive and lethal form of brain cancer. Despite recent advances in diagnostics and treatments, prognosis for advanced patients suffering from these diseases remains poor. Therefore, identification of new therapeutic targets for glioma is of significant importance. In this study, we identified the important role of Smad interacting protein 1 (SIP1; also known as ZEB2) in glioma. We firstly found that SIP1 expression was high in four tumorigenic glioma cell lines but low in two nontumorigenic glioma cell lines. By knockdown or overexpression assay, we discovered that knockdown of SIP1 expression statistically significantly inhibited cell migration and invasion of tumorigenic glioma cells, while overexpression of SIP1 promoted cell migration and invasion of nontumorigenic glioma cells. SIP1 knockdown inhibits and overexpression promotes glioma cell clonogenicity in vitro. Further studies identified that SIP1 overexpression inhibits expression of E-cadherin and enhances expression of mesenchymal proteins such as fibronectin and vimentin. This study supports the rationale for developing SIP1 as a potential therapeutic and diagnostic target for gliomas. PMID- 19806325 TI - [Apparent saving through imitations of ready-made dermatologic agents]. PMID- 19806324 TI - [Medical information systems in the internet : current data about the dermatologic web site www.dermis.net]. AB - www.dermis.net was established in 1994 as a dermatological information system with now more than 4,000 pages and over 7,500 dermatological images in an atlas. 1,200 dermatological diagnoses and medical information with 1,100 synonyms can be searched by body location and in alphabetical order. Additionally, there are seven information modules about skin cancer, atopic dermatitis, rosacea, skin care, eczema, fungal diseases and hemorrhoids with links to other medical pages like Cochrane Library. Pediatric skin diseases are covered in PeDOIA. www.dermis.net is meanwhile available in German, English, Spanish, Portuguese, French and Turkish language. Current contents of this information system and user behavior are presented in this paper. PMID- 19806323 TI - Microarray analysis of the moss Physcomitrella patens reveals evolutionarily conserved transcriptional regulation of salt stress and abscisic acid signalling. AB - Regulatory networks of salt stress and abscisic acid (ABA) responses have previously been analyzed in seed plants. Here, we report microarray expression profiles of 439 genes encoding transcription-associated proteins (TAPs) in response to salt stress and ABA in the salt-tolerant moss Physcomitrella patens. Fourteen and 56 TAP genes were differentially expressed within 60 min of NaCl and ABA treatment, respectively, indicating that these responses are regulated at the transcriptional level. Overlapping expression profiles, as well as the up regulation of ABA biosynthesis genes, suggest that ABA mediates the salt stress responses in P. patens. Comparison to public gene expression data of Arabidopsis thaliana and phylogenetic analyses suggest that the role of DREB-like, Dof, and bHLH TAPs in salt stress responses have been conserved during embryophyte evolution, and that the function of ABI3-like, bZIP, HAP3, and CO-like TAPs in seed development and flowering emerged from pre-existing ABA and light signalling pathways. PMID- 19806326 TI - [Pro- and prebiotics in dermatology]. PMID- 19806327 TI - [Oligoclonality of multiple primary melanoma]. PMID- 19806328 TI - Effects of artemisinin derivative on the growth metabolism of Tetrahymena thermophila BF5 based on expression of thermokinetics. AB - The toxic effects of artesunate and dihydroartemisinin on the growth metabolism of Tetrahymena thermophila BF5 were studied by microcalorimetry. The results showed that: (1) low concentrations of artesunate ( or = 4 pN+ and received RT had comparable outcomes after 15 years. The 15-year OS in the subgroup with ME and < or = 3 pN+ nodes was 57.0% and 46.6% (p = 0.0004) with RT (UZ Brussel) and without RT (SEER), respectively. For BCS and < or = 3 pN+, the same significant difference in OS at 15 years was seen: 63.8% after RT (UZ Brussel) and 60.4% without RT (SEER; p = 0.0029). CONCLUSION: RT provides a survival benefit in patients with < or = 3 or > or = 4 pN+; the indication for postoperative RT should therefore be adapted in future consensus meetings. PMID- 19806331 TI - Medial tumor localization in breast cancer--an unappreciated risk factor? AB - PURPOSE: To demonstrate the unfavorable results in survival rates in patients with medial breast cancer compared to patients with laterally located tumors of the mammary gland. PATIENTS AND METHODS: Between 1984 and 1995, 1,089 patients presenting with a total of 1,100 pT1-2 invasive carcinomas of the breast were treated at the authors' institution. 707 presented with tumors in the lateral quadrants, 294 with tumors in the medial quadrants, and 99 with tumors in the central quadrant. Treatment protocols involved breast-conserving surgery and whole-breast radiotherapy in all women, followed by a tumor bed boost dose according to risk factors for local recurrence. All axillary node-positive patients underwent systemic therapy (six cycles of classic CMF and/or 2-5 years of tamoxifen 20 mg/day). Rates of actuarial survival and local control were calculated by the Kaplan-Meier method and differences in survival curves were compared by use of the log-rank test. RESULTS: The mean follow-up of survivors was 97 months (range 36-192 months). Comparing patients with medial and lateral tumors, the actuarial survival data were significantly better for patients with lateral tumors. At 10 years, overall survival for patients with medial tumors was 71%, for patients with lateral tumors 81.8% (p < 0.025), disease-specific survival for patients with medial tumors 79.9%, for patients with lateral tumors 89.1% (p < 0.025). There was no significant difference in local tumor control according to tumor location. CONCLUSION: Medial tumor location is associated with a lower survival rate, but not with inferior local tumor control. Failure to identify nodal metastases confined to the internal mammary chain may lead to undertreatment with systemic/local agents and compromised survival. PMID- 19806332 TI - Immediate reconstruction with an expander/implant following ablatio mammae because of breast cancer : side effects and cosmetic results after adjuvant chest wall radiotherapy. AB - BACKGROUND AND PURPOSE: Timing and sequencing of radiotherapy in the context of allogenous breast reconstruction have not been standardized. The aim of the present study was to assess the influence of adjuvant radiotherapy on morbidity and patient satisfaction after allogenous breast reconstruction. PATIENTS AND METHODS: 33 patients underwent mastectomy between 1999 and 2008 and had immediate breast reconstruction with an expander placement in subpectoral/epipectoral location. 24 patients had adjuvant chemotherapy. Adjuvant external-beam radiotherapy with a median dose of 50.4 Gy was given after expander filling and on average 5.2 months prior to placement of the definitive implant. 22 patients with the definitive implant were considered for analysis of capsular fibrosis rate. Questionnaires were sent to all patients to assess cosmetic outcome and satisfaction. RESULTS: Acute adverse effects were comparable to adjuvant radiotherapy after breast-conserving surgery, resulting in an erythema rate grade 1/2/3 in 21.2%/66.7%/6.1% of patients, respectively. After a mean follow-up of 24.9 months, 9.1%/18.2%/15.2%/9.1% of patients presented a capsular fibrosis grade 1/2/3/4, respectively. Severe deformation/asymmetry of the reconstructed breast was seen in 27.3%/33.3% of patients, respectively. Of the 22 patients with definitive implant, five (22.7%) lost the implant due to painful capsular fibrosis. Of these 22 patients, 50% were very satisfied or satisfied with the reconstruction result. Overall, 81% of patients would request breast reconstruction again. CONCLUSION: Adjuvant radiotherapy with the use of a subtotally filled expander prior to definitive allogenous breast reconstruction is feasible with acceptable morbidity. An interdisciplinary consultation concerning the cosmetic outcome and potential side effects is absolutely necessary. PMID- 19806333 TI - Radiochemotherapy including cisplatin alone versus cisplatin + 5-fluorouracil for locally advanced unresectable stage IV squamous cell carcinoma of the head and neck. AB - BACKGROUND AND PURPOSE: The optimal radiochemotherapy regimen for advanced head and-neck cancer is still debated. This nonrandomized study compares two cisplatin based radiochemotherapy regimens in 128 patients with locally advanced unresectable stage IV squamous cell carcinoma of the head and neck (SCCHN). PATIENTS AND METHODS: Concurrent chemotherapy consisted of either two courses cisplatin (20 mg/m(2)/d1-5 + 29-33; n = 54) or two courses cisplatin (20 mg/m(2)/d1-5 + 29-33) + 5-fluorouracil (5-FU; 600 mg/m(2)/d1-5 + 29-33; n = 74). RESULTS: At least one grade 3 toxicity occurred in 25 of 54 patients (46%) receiving cisplatin alone and in 52 of 74 patients (70%) receiving cisplatin + 5 FU. The latter regimen was particularly associated with increased rates of mucositis (p = 0.027) and acute skin toxicity (p = 0.001). Seven of 54 (13%) and 20 of 74 patients (27%) received only one chemotherapy course due to treatment related acute toxicity. Late toxicity in terms of xerostomia, neck fibrosis, skin toxicity, and lymphedema was not significantly different. The 2-year locoregional control rates were 67% after cisplatin alone and 52% after cisplatin + 5-FU (p = 0.35). The metastases-free survival rates were 79% and 69%, respectively (p = 0.65), and the overall survival rates 70% and 51%, respectively (p = 0.10). On multivariate analysis, outcome was significantly associated with performance status, T-category, N-category, hemoglobin level prior to radiotherapy, and radiotherapy break > 1 week. CONCLUSION: Two courses of fractionated cisplatin (20 mg/m(2)/day) alone appear preferable, as this regimen resulted in similar outcome and late toxicity as two courses of cisplatin + 5-FU, but in significantly less acute toxicity. PMID- 19806334 TI - Chemoradiotherapy with weekly cisplatin 40 mg/m(2) in 103 head-and-neck cancer patients: a cumulative dose-effect analysis. AB - BACKGROUND AND PURPOSE: In patients with head-and-neck cancer treated with chemoradiotherapy (CRT), a cisplatin-based regimen is often used. Several treatment schedules are accepted with a cumulative cisplatin dose of 200 mg/m(2) (CisCD200) given during radiotherapy. The aim of this analysis was to investigate feasibility and efficacy of a weekly cisplatin 40 mg/m(2) regimen. PATIENTS AND METHODS: During 08/2001 and 12/2006, 103 patients with squamous head-and-neck cancer received concurrent CRT with intended weekly cisplatin 40 mg/m(2) and were analyzed retrospectively. CRT was definitive for a newly diagnosed primary in 62, postoperative in 16, and for recurrence in 25 patients. Most patients had carcinoma of the hypo- and oropharynx (81%). Patients received a median total dose of 70 Gy (range, 42-71.2 Gy). RESULTS: Only 42 patients (41%) received a CisCD200 predominantly due to hematotoxicity. Actuarial 12- and 18-month overall survival (OS) for patients with and without CisCD200 was 83.3% versus 72.1% (p = 0.19) and 66.7% versus 67.2% (p = 0.86), the 12- and 18-month locoregional control (LRC) 66.7% versus 78.7% (p = 0.325) and 59.5% versus 78.7% (p = 0.109), respectively. Multivariate analysis revealed only type of CRT (definitive vs. recurrent) and T-classification as significant variables predicting OS and LRC. CONCLUSION: Feasibility and efficacy of CRT with weekly cisplatin 40 mg/m2 were suboptimal in this analysis. However, the prospects of weekly cisplatin may be its more suitable integration into emerging trimodality concepts combining CRT with molecularly targeted agents. PMID- 19806335 TI - Exposure of treating physician to radiation during prostate brachytherapy using iodine-125 seeds: dose measurements on both hands with thermoluminescence dosimeters. AB - BACKGROUND AND PURPOSE: Only sparse reports have been made about radiation exposure of the treating physician during prostate seed implantation. Therefore, thermoluminescence dosimeter (TLD) measurements on the index fingers and the backs of both hands were conducted. MATERIAL AND METHODS: Stranded iodine-125 seeds with a mean apparent activity of 27.4 MBq per seed were used. During application, the treating physician manipulated the loaded needle with the index fingers, partially under fluoroscopic control. Four physicians with varying experience treated 24 patients. The radiation exposure was determined with TLD 100 chips attached to the index fingertips and the backs of hands. Radiation exposure was correlated with the physician's experience. RESULTS: The average brachytherapy duration by the most experienced physician was 19.2 min (standard deviation sigma = 1.2 min; novices: 34.8 min [sigma = 10.2 min]). The mean activity was 1,703 MBq (sigma = 123 MBq), applied with 16.3 needles (sigma = 2.5 needles; novices: 1,469 MBq [sigma = 229 MBq]; 16.8 needles [sigma = 2.3 needles]). The exposure of the finger of the "active hand" and the back of the hand amounted to 1.31 mSv (sigma = 0.54 mSv) and 0.61 mSv (sigma = 0.23 mSv), respectively (novices: 2.07 mSv [sigma = 0.86 mSv] and 1.05 mSv [sigma = 0.53 mSv]). CONCLUSION: If no other radiation exposure needs to be considered, an experienced physician can perform about 400 applications per year without exceeding the limit of 500 mSv/year; for novices, the corresponding figure is about 200. PMID- 19806336 TI - The influence of different IMRT techniques on the peripheral dose: a comparison between sMLM-IMRT and helical tomotherapy. AB - PURPOSE: To investigate how segmented multileaf modulation-(sMLM-)based intensity modulated radiotherapy (IMRT) and dynamic helical tomotherapy (ToTh) affect the peripheral dose (PD) outside the treated region. MATERIAL AND METHODS: A cuboid Perspex phantom was scanned in a computed tomograph. Different artificial cases were contoured consisting of OARs surrounded by cylindrically shaped planning target volumes (PTVs) with different dimensions. Radiotherapy plans were generated with the sMLM system Konrad (Siemens) and with the ToTh planning system. The plans were optimized in such a way that the dose-volume histograms showed comparable results. The sMLM plans were applied with a linac Primus (Siemens OCS), the ToTh plans with the HiArt system (TomoTherapy); both with 6 MV. Measurements of PDs were performed along the longitudinal axis of the phantom outside the primary beam at different distances from the edge of the PTV (horizontal PD) and also at different depths at a fixed distance from the isocenter (vertical PD). Additional experiments to separate the scatter dose caused by the phantom were performed. This was realized by removing the part of the phantom lying in the primary beam, then applying the same plans like before. RESULTS: All PD values were normalized to the median dose of the PTV. The PD values for the different PTVs decrease with decreasing PTV size. They also decrease with increasing distance from the isocenter. The horizontal values are in a range of 7% for the largest PTV (diameter = 15 cm) near the primary dose region to 0.2% for the smallest PTV (diameter = 5 cm) far from the primary dose region. The ToTh values are higher than the sMLM values by a maximal factor of 2 near the primary dose region. They become more similar with increasing distance from the edge of the PTV in longitudinal direction. The PD values are nearly equal at a distance of 25 cm from the edge of the PTV. The vertical PDs are higher for the ToTh at depths of > 1 cm but higher for sMLM close to the surface. By removing the scatter cube, the horizontal PD values at middle distances are reduced to one third of the PD values with scatter cube for ToTh (0.5%) and to one half for sMLM (0.8%). This means that without scatter cube the PD for ToTh is lower than that for sMLM. The measured PD values without scatter cube are in the same dimension as published data. CONCLUSION: The increasing PDs and their trend with increasing PTV size can be explained by Compton scattering of photons from the irradiated volume toward the off-axis measuring points. The further increase of the PD in case of ToTh relative to sMLM is not easy to explain. Different presumptions are possible. The larger field length (in longitudinal direction) of the ToTh plans (consisting of the "real" field length and the overlap) relative to the sMLM plans could be one reason for the higher PD values. The softer energy spectrum of the HiArt machine with more sideward Compton scattering contributions could be another reason. PMID- 19806339 TI - [Shaking trauma and baby syndrome]. PMID- 19806338 TI - TCF7L2 genetic variants and progression to diabetes in the Chinese population: pleiotropic effects on insulin secretion and insulin resistance. AB - TCF7L2 genetic variants were associated with progression to type 2 diabetes in Europeans. However, the role of TCF7L2 in type 2 diabetes remained uncertain in Chinese. Seventeen tag single nucleotide polymorphisms were genotyped in 1,094 subjects of Chinese origin from the Stanford Asia-Pacific Program for Hypertension and Insulin Resistance family study. At baseline, the rs7903146 T allele in the exon 4 linkage disequilibrium (LD) block were associated with lower insulinogenic index at 60 min (P = 0.01), while the rs290481 G allele near the 3' end was associated with higher 2-h post-challenge glucose (P = 0.003) and insulin concentration (P = 0.02), elevated systolic (P = 0.01) and diastolic blood pressure (P = 0.006), lower waist circumference (P = 0.01), and increased steady state plasma glucose (SSPG) concentration measured with modified insulin suppression test (P = 0.02). Over an average follow-up period of 5.43 years, participants with the rs7903146 T allele or variants in the same LD block, but not those with the rs290481 G allele, were more likely to progress to diabetes (hazard ratio = 2.61, 95% confidence interval, 1.27-5.39, P = 0.009) than were non-carriers. TCF7L2 gene expression was inversely associated with SSPG in human visceral (r = -0.73, P = 0.006) and subcutaneous adipose tissue (r = -0.62, P = 0.03). TCF7L2 may exert pleiotropic effects on insulin secretion or insulin resistance. However, only variants associated with impaired beta-cell function predict progression to diabetes in Chinese. PMID- 19806340 TI - [Multiple space-occupying lesions of the liver: a rare differential diagnosis]. AB - The case of a 73-year-old female patient with a known increase in tumor markers is described. The course of this patient shows that a definitive diagnosis can sometimes not be achieved with comprehensive laboratory and imaging investigations alone. PMID- 19806341 TI - [Cranial birth trauma]. AB - Injuries to an infant that result during the birth process are categorized as birth trauma. Cranial injuries due to mechanical forces such as compression or traction include caput succedaneum, cephalhematoma, subgaleal hematoma and intracranial hemorrhaging. Hypoxic ischemic encephalopathy is the consequence of systemic asphyxia occurring during birth. PMID- 19806342 TI - [Systemic therapy of metastatic renal cell carcinoma: from many options to the therapeutic strategy]. AB - In the last 5 years the paradigms for the treatment of metastatic renal cell cancer have fundamentally changed. Until 2005 systemic therapy was limited to the immunomodulating cytokines interferon-alfa and interleukin-2, in recent years, however, tyrosine kinase inhibitors, mTor inhibitors and monoclonal antibodies have been established for this therapeutic situation. Without validated predictive biomarkers it is currently not possible to select patients who are likely to benefit from a certain therapy. Therefore, most current guidelines stratify the patients into risk groups according to the MSKCC risk score. The resulting treatment algorithm for first-line therapy is limited to these new drugs within all risk groups. Since approval for more tyrosine kinase inhibitors and mTOR inhibitors is currently awaited, the number of treatment options will expand further in the near future. The present paper reviews the present study data and aims to provide practical advice for the treatment of patients suffering from metastatic renal cell cancer. PMID- 19806343 TI - Mechanisms of the inhibition of endplate acetylcholine receptors by antiseptic chlorhexidine (experiments and models). AB - Mechanisms of the inhibition of evoked multiquantal endplate currents (EPC) by chlorhexidine (CHX) were studied in electrophysiological experiments and by mathematical modeling to discriminate between possible channel, receptor, and non receptor effects of this common antiseptic drug. Experiments were carried out on the isolated neuromuscular preparation of the cut m. sartorius of the frog Rana ridibunda. The nerve-stimulation-evoked endplate currents were measured by standard double microelectrode technique. For the mathematical simulation, a method based on the solution of a system of ordinary differential equations was used. CHX in milimolar concentrations suppressed the amplitude and shortened the evoked EPC. Recovery of the EPC amplitude was very slow, and EPC shortening persisted during 30-40 min washout of the drug. There is no indication that CHX competes for acetylcholine or carbachol binding site(s). A comparison of the experimental data with mathematical simulation made it possible to construct a reliable kinetic scheme, which describes the action of CHX. CHX induces a combined slow blockade of the open ionic channel and long-lasting allosteric inhibition of the nicotinic acetylcholine receptor. The very slow washout of the drug in terms of EPC amplitude and virtually no recovery of the shortened EPC time course might substantiate certain caution to avoid unintentional high-dose application during its antibacterial application. PMID- 19806345 TI - Proteomic strategies for the identification of proteinaceous binders in paintings. AB - The identification of proteinaceous components in paintings remains a challenging task for several reasons. In addition to the minute amount of sample available, complex and variable chemical composition of the paints themselves, possible simultaneous presence of several binders and contaminants, and degradation of the original materials due to aging and pollution are complicating factors. We proposed proteomic strategies for the identification of proteins in binders of paintings that can be adapted to overcome the requirements and difficulties presented by specific samples. In particular, we worked on (1) the development of a minimally invasive method based on the direct tryptic cleavage of the sample without protein extraction; (2) the use of microwave to enhance the enzymatic digestion yield, followed by the analysis of the peptide mixtures by nanoLC-MS/MS with electrospray ionization (ESI). Moreover, as an additional tool to tackle the problem of contaminating proteins, we exploited the possibility of generating an exclusion list of the mass signals that in a first run had been fragmented and that the mass spectrometer had to ignore for fragmentation in a subsequent run. The methods, tested on model samples, allowed the identification of milk proteins in a sample from paintings attributed to Cimabue and Giotto, thirteenth-century Italian masters, decorating the vaults of the upper church in the Basilica of St. Francis in Assisi, Italy. PMID- 19806344 TI - Nicotine withdrawal and kappa-opioid receptors. AB - RATIONALE: The synthesis and release of dynorphin are increased in the caudate/putamen (CPU) and nucleus accumbens (NAc) of nicotine-withdrawn mice, suggesting a role in the nicotine abstinence syndrome. OBJECTIVES: This study aims to investigate the consequences of enhanced dynorphinergic activity on kappa opioid receptor (KOPr) expression, coupling, and function in CPU and NAc following chronic nicotine administration and withdrawal. METHODS: Mice were injected with nicotine-free base 2 mg/kg, or saline, sc, four times daily for 14 days and experiments performed at 24, 48, and 72 h after drug discontinuation. KOPr binding and mRNA were evaluated by [(3)H]-U69,593 autoradiography and in situ hybridization. KOPr coupling and function were investigated by agonist (U69 593)-stimulated [(35)S]GTPgammaS binding autoradiography and inhibition of adenylyl cyclase activity. RESULTS: KOPr binding density and mRNA in CPU and NAc were unaltered during nicotine withdrawal; however, KPOr mRNA was increased in midbrain. U69,593-stimulated [(35)S]GTPgammaS binding was attenuated in both striatal regions, especially in NAc. In NAc shell and core, stimulated [(35)S]GTPgammaS binding was significantly decreased by 24 h and further declined over the 72 h observation period. In CPU, significant changes were observed only at 72 h. Basal adenylyl cyclase activity decreased early during nicotine withdrawal and recovered by 48 h. Stimulation with U69,593 failed to inhibit adenylyl cyclase activity at all times studied. CONCLUSIONS: These observations suggest that KOPr coupling and function are impaired in NAc and CPU during nicotine withdrawal, and imply receptor desensitization. KOPr desensitization might be a mechanism to ameliorate aversive behavioral symptoms, as nicotine withdrawal evolves. PMID- 19806346 TI - Matrix solid-phase dispersion and solid-phase microextraction applied to study the distribution of fenbutatin oxide in grapes and white wine. AB - The fate of the acaricide fenbutatin oxide (FBTO) during the elaboration of white wine is evaluated. Matrix solid-phase dispersion (MSPD) and solid-phase microextraction (SPME) were used as sample preparation techniques applied to the semi-solid and the liquid matrices involved in this research, respectively. Selective determination of FBTO was achieved by gas chromatography with atomic emission detection (GC-AED). GC coupled to mass spectrometry was also used to establish the identity of FBTO by-products detected in must and wine samples. MSPD extractions were accomplished using C18 as dispersant and co-sorbent. Sugars and other polar interferences were first removed with water and water/acetone mixtures, then FBTO was recovered with 8 mL of acetone. When used in combination with GC-AED, the MSPD method provided limits of quantification (LOQs) in the low nanogram per gram range, recoveries around 90% and relative standard deviations below 13% for extractions performed in different days. Performance of SPME for must and wine was mainly controlled by the extraction temperature, time and fibre coating. Under final conditions, FBTO was extracted in the headspace mode for 45 min at 100 degrees C, using a 100 microm poly(dimethylsiloxane)-coated fibre. The achieved LOQs remained around or below 0.1 ng mL(-1), depending on the type of sample, and the inter-day precision ranged from 10% to 13%. FBTO residues in grapes stayed mostly on the skin of the fruit. Although FBTO was not removed during must and white wine elaboration, it remained associated with suspended particles existing in must and lees, settled after must fermentation, with a negligible risk of being transferred to commercialised wine. On the other hand, two by-products of FBTO (bis and mono (2-methyl-2-phenylpropyl) tin) were identified, for first time, in must and final white wines obtained from FBTO treated grapes. Found values for the first species ranged from 0.03 to 0.9 ng mL( 1). PMID- 19806347 TI - Improved rapid assay of plasma uric acid by short-end injection capillary zone electrophoresis. AB - A rapid and simple short-end injection capillary zone electrophoresis method was developed for the quantification of plasma uric acid. The separation was performed in an uncoated fused-silica capillary (50 microm ID, 60 cm total length, 10.2 cm effective length) by using as a background electrolyte a 75 mmol/L glycylglycine solution titrated with NaOH 5 mol/L to pH 9.0, a voltage of 28 kV, a cartridge temperature of 15 degrees C, and direct UV detection at 292 nm. Under optimized conditions, uric acid was determinate in little more than 1 min (1.076 minutes). In order to verify the accuracy of the analysis, urate levels were measured in 543 apparently healthy volunteers by the new assay and our previous method, and the obtained data were compared by Passing-Bablock regression, Bland-Altman test, and a new regression-based approach, which showed a good agreement between two methods. PMID- 19806348 TI - Determination of ketamine and amphetamines in hair by LC/MS/MS. AB - A liquid chromatography-tandem mass spectrometry method was developed for the determination of ketamine (with its metabolite norketamine) and some amphetamines (amphetamine, methamphetamine, methylenedioxyamphetamine, and 3,4 methylenedioxymethamphetamine). This method was developed to determine these compounds in hair and is able to simultaneously quantify all of them in human hair. Hair samples (20 mg) were washed and pulverized, and an extraction with formic acid (0.01%) and ultrasonication for 4 h was used. Deuterated analogs of the analytes were used as internal standards for quantification. Linearity from 0.5 to 25 ng/mg was obtained for both ketamine (and norketamine) and amphetamines with correlation coefficients exceeding 0.99. The limit of detection and the limit of quantification obtained were 0.1 and 0.5 ng/mg, respectively, for ketamine and amphetamines. A total of 25 hair samples from known drug abusers (relating to designer drug consumption or consumption of amphetamines) were examined by this validated method. The results show that the proposed method is suitable for testing these drugs in a single sample of hair. In addition, it is simpler and faster than analysis by conventional methods such as gas chromatography-mass spectrometry, which usually require a more laborious extraction procedure and, in most of cases, an additional derivatization process. PMID- 19806349 TI - Identifying cytoplasmic proteins that affect receptor clustering using fluorescence resonance energy transfer and RNA interference. AB - Unraveling the complex, dynamic organization of the cell membrane can provide vital information about many aspects of cellular functions. Reported herein is a method for identifying cytoplasmic proteins that affect cell membrane protein organization. RNA interference (RNAi) is used to reduce the expression of select cytoplasmic proteins and a fluorescence resonance energy transfer (FRET) assay is used to measure changes in receptor microclustering. The advantage of this assay is that it does not require attaching fluorescent tags to the receptor. A change in energy transfer after reducing the expression of a cytoplasmic protein provides information about the protein's role in altering receptor organization. As a demonstration of the method, cytoplasmic proteins involved in integrin microclustering have been identified. The cytoplasmic proteins targeted in this study include: dreadlock, integrin-linked kinase, paxillin, steamer duck, vinculin, rhea, focal adhesion kinase, and actin 42A. Reducing the expression of vinculin, paxillin, rhea, and focal adhesion kinase increased integrin microclustering, as measured by an increase in energy transfer in cells expressing alphaPS2CbetaPS integrins. No change in integrin microclustering was measured in a control cell line. Integrin mutants exhibited different microclustering properties compared to the wild-type integrins after reducing the expression of the listed cytoplasmic proteins. The results demonstrate the utility of this assay format, and provide insight into the function of cytoplasmic proteins in integrin microclustering. PMID- 19806350 TI - Static and dynamic discharge properties of vestibular-nerve afferents in the mouse are affected by core body temperature. AB - The goal of this study was to determine the effect of changes in core body temperature on the resting discharge rate and sensitivity of vestibular-nerve afferents. Extracellular recordings were made from vestibular-nerve afferents innervating the semicircular canals in anesthetized C57BL/6 mice maintained at a core body temperature of either 30-32 degrees C (T (31)) or 35-37 degrees C (T (36)). The resting rates of regular (CV* < 0.1) and irregular afferents (CV* > 0.1) were lower at T (31) than at T (36). Sensitivity and phase were compared for rotations ranging from 0.1 to 12 Hz by calculating coefficients of a transfer function, g . t(c)S . (t(z)S +1)/(t(c)S + 1), for each afferent. The sensitivity (g) increased with CV* and with higher core body temperature. The value of the coefficient representing the low-frequency dynamics (t (c)) varied inversely with CV* but did not change with core body temperature. The high-frequency dynamics represented by t (z) increased with CV* and decreased with higher core body temperature. These findings indicate that changes in temperature have effects on the static and dynamic properties of vestibular-nerve afferents. PMID- 19806351 TI - An enhanced voxel-based morphometry method to investigate structural changes: application to Alzheimer's disease. AB - INTRODUCTION: When characterizing regional cerebral gray matter differences in structural magnetic resonance images (sMRI) by voxel-based morphometry (VBM), one faces a known drawback of VBM, namely that histogram unequalization in the intensity images introduces false-positive results. METHODS: To overcome this limitation, we propose to improve VBM by a new approach (called eVBM for enhanced VBM) that takes the histogram distribution of the sMRI into account by adding a histogram equalization step within the VBM procedure. Combining this technique with two most widely used VBM software packages (FSL and SPM), we studied GM variability in a group of 62 patients with Alzheimer's disease compared to 73 age matched elderly controls. RESULTS: The results show that eVBM can reduce the number of false-positive differences in gray matter concentration. CONCLUSION: Because it takes advantage of the properties of VBM while improving sMRI histogram distribution at the same time, the proposed method is a powerful approach for analyzing gray matter differences in sMRI and may be of value in the investigation of sMRI gray and white matter abnormalities in a variety of brain diseases. PMID- 19806352 TI - Infantile chylothorax associated with staphylococcal paravertebral discitis. AB - We report an unusual case of chylothorax in an infant associated with a staphylococcal paravertebral abscess secondary to discitis affecting the thoracic spine. We discuss the initial presentation with sepsis and chylothorax and the delayed diagnosis of discitis. We also discuss the imaging features and treatment in this case. PMID- 19806353 TI - On computational approaches for size-and-shape distributions from sedimentation velocity analytical ultracentrifugation. AB - Sedimentation velocity analytical ultracentrifugation has become a very popular technique to study size distributions and interactions of macromolecules. Recently, a method termed two-dimensional spectrum analysis (2DSA) for the determination of size-and-shape distributions was described by Demeler and colleagues (Eur Biophys J 2009). It is based on novel ideas conceived for fitting the integral equations of the size-and-shape distribution to experimental data, illustrated with an example but provided without proof of the principle of the algorithm. In the present work, we examine the 2DSA algorithm by comparison with the mathematical reference frame and simple well-known numerical concepts for solving Fredholm integral equations, and test the key assumptions underlying the 2DSA method in an example application. While the 2DSA appears computationally excessively wasteful, key elements also appear to be in conflict with mathematical results. This raises doubts about the correctness of the results from 2DSA analysis. PMID- 19806354 TI - A novel acid-stable, acid-active beta-galactosidase potentially suited to the alleviation of lactose intolerance. AB - Extracellular beta-galactosidase produced by a strain of Aspergillus niger van Tiegh was purified to homogeneity using a combination of gel filtration, ion exchange, chromatofocusing, and hydrophobic interaction chromatographies. The enzyme displayed a temperature optimum of 65 degrees C and a low pH optimum of between 2.0 and 4.0. The monomeric glycosylated enzyme displayed a molecular mass of 129 kDa and an isoelectric point of 4.7. Protein database similarity searching using mass spectrometry-derived sequence data indicate that the enzyme shares homology with a previously sequenced A. niger beta-galactosidase. Unlike currently commercialised products, the enzyme displayed a high level of stability when exposed to simulated gastric conditions in vitro, retaining 68+/-2% of original activity levels. This acid-stable, acid-active beta-galactosidase was formulated, along with a neutral beta-galactosidase from Kluyveromyces marxianus DSM5418, in a novel two-segment capsule system designed to ensure delivery of enzymes of appropriate physicochemical properties to both stomach and small intestine. When subjected to simulated full digestive tract conditions, the twin lactase-containing capsule hydrolyzed, per unit activity, some 3.5-fold more lactose than did the commercial supplemental enzyme. The acid-stable, acid-active enzyme, along with the novel two-segment delivery system, may prove beneficial in the more effective treatment of lactose intolerance. PMID- 19806355 TI - An extensive analysis of the hereditary hemochromatosis gene HFE and neighboring histone genes: associations with childhood leukemia. AB - The most common mutation of the HFE gene C282Y has shown a risk association with childhood acute lymphoblastic leukemia (ALL) in Welsh and Scottish case-control studies. This finding has not been replicated outside Britain. Here, we present a thorough analysis of the HFE gene in a panel of HLA homozygous reference cell lines and in the original population sample from South Wales (117 childhood ALL cases and 414 newborn controls). The 21 of 24 variants analyzed were from the HFE gene region extending 52 kb from the histone gene HIST1H1C to HIST1H1T. We identified the single-nucleotide polymorphism (SNP) rs807212 as a tagging SNP for the most common HFE region haplotype, which contains wild-type alleles of all HFE variants examined. This intergenic SNP rs807212 yielded a strong male-specific protective association (per allele OR = 0.38, 95% CI = 0.22-0.64, P (trend) = 0.0002; P = 0.48 in females), which accounted for the original C282Y risk association. In the HapMap project data, rs807212 was in strong linkage disequilibrium with 25 other SNPs spanning 151 kb around HFE. Minor alleles of these 26 SNPs characterized the most common haplotype for the HFE region, which lacked all disease-associated HFE variants. The HapMap data suggested positive selection in this region even in populations where the HFE C282Y mutation is absent. These results have implications for the sex-specific associations observed in this region and suggest the inclusion of rs807212 in future studies of the HFE gene and the extended HLA class I region. PMID- 19806356 TI - Association of homocysteine, asymmetric dimethylarginine, and nitric oxide with preeclampsia. AB - PURPOSE: Endothelial dysfunction underlies the pathogenesis of preeclampsia, but its mechanism has not yet been completely understood. In this study we have aimed to measure homocysteine (Hcy), asymmetric dimethylarginine (ADMA), and nitric oxide (NO) levels as endothelial dysfunction markers in preeclamptic women. METHODS: Control-case study with 62 preeclamptic patients and 30 controls without pregnancy complications was conducted. Plasma total Hcy, determined by capillary column gas chromatography/mass spectrometry (GC/MS), was correlated with serum ADMA (determined by liquid chromatography/tandem mass spectrometry using (13)C(6) L: -arginine as the internal standard) and NO (analyzed by GC/MS). RESULTS: There was a highly significant increase in the plasma concentration of homocysteine (P < 0.001) and ADMA (P < 0.001) and a highly significant decrease in the plasma concentration of nitric oxide (P < 0.001) among the preeclamptic patients. The differences were more significant between mild and severe preeclampsia, with and without eclampsia, with and without HELLP (hemolysis, elevated serum level of liver enzymes, and low platelets). In the combined patients and control groups a highly significant positive correlation was found between the plasma concentrations of homocysteine and ADMA (r = 0.853, P < 0.001). In addition, significant negative correlations were detected between the plasma concentrations of nitric oxide and the plasma concentration of homocysteine (r = -0.870, P < 0.001) and ADMA (r = -0.895, P < 0.001). These significant correlations were found to persist, even when they were restricted to the preeclamptic patients. CONCLUSIONS: The homocysteine-ADMA-NO may be at least partly responsible for etiology in preeclampsia and could be regarded as markers for the severity of the disease. Therefore, L: -arginine may represent a novel therapy for the treatment of preeclampsia. PMID- 19806357 TI - The effect of interferon beta-1a on optic neuritis relapse in patients with multiple sclerosis. AB - BACKGROUND: To evaluate the clinical effect of interferon beta-1a on optic neuritis (ON) relapse in patients with multiple sclerosis (MS) in Taiwan. METHODS: Data were collected from 23 MS patients with ON at National Taiwan University Hospital between January 1, 1993 and February 1, 2007. Twenty-three MS patients with ON received interferon beta-1a (Rebif) 44 microg via subcutaneous injection three times weekly. All patients received corticosteroids pulse therapy followed by oral prednisolone for acute ON. The annual relapse rate (ARR) of ON in these MS patients before and after the use of interferon beta-1a (Rebif) was the main clinical parameter of outcome in this study. RESULTS: The ARR of ON was lower in the posttreatment period than in the pretreatment period (P = 0.0068). Thirteen patients (56.5%) had improved final visual acuity (>2 lines), and the other ten patients (43.5%) had stable final visual outcome (-2 lines < X < 2 lines). In addition, no recurrence of ON was noted in 15 patients (65.2%) during the posttreatment period. CONCLUSIONS: The use of interferon beta-1a 44 microg via subcutaneous injection three times weekly did not increase the ON attacks in MS patients receiving this treatment. In addition, beneficial effects were found with the use of interferon beta-1a on these patients. PMID- 19806358 TI - Longitudinal relationship between physical activity and cardiometabolic factors in overweight and obese adults. AB - Few studies have reported longitudinal relationships between physical activity (PA) and cardiometabolic risk factors over time using repeated assessments in overweight or obese adults. We conducted a longitudinal study in 127 participants (81% with body mass index > 30 kg/m(2)) who completed a 12-month behavioral intervention for weight loss between 2003 and 2005 in Pittsburgh, PA, USA. Using absolute change scores from baseline to each time point (i.e., 6 and 12 months) for all studied variables (Delta = time point - baseline), we performed mixed effects modeling to examine relationships between PA and cardiometabolic risk factors, after adjusting for body weight, energy intake and other covariates (i.e., age, gender, and ethnicity). PA was assessed as energy expenditure (kcal/week) using the Paffenbarger activity questionnaire. Over the 12-month period, energy expenditure increased (Delta1,370 kcal/week at 6 months vs. Delta886 kcal/week at 12 months); body weight decreased (Delta8.9 kg at 6 months vs. Delta8.4 kg at 12 months). The average increase in energy expenditure over 12 months was significantly and independently related to reductions in total cholesterol (F = 6.25, p = 0.013), low-density lipoprotein cholesterol (LDL-C) (F = 5.08, p = 0.025) and fasting blood glucose (F = 5.10, p = 0.025), but not to other risk factors (i.e., fasting insulin, high-density lipoprotein cholesterol, triglycerides, and waist circumference). In conclusion, among overweight and obese adults undergoing a weight loss intervention, increased energy expenditure over 12 months may improve total cholesterol and LDL-C, important coronary risk factors, and fasting blood glucose, a metabolic risk factor. PMID- 19806359 TI - The importance of serine 161 in the sodium channel beta3 subunit for modulation of Na(V)1.2 gating. AB - Voltage-gated sodium (Na) channels contribute to the regulation of cellular excitability due to their role in the generation and propagation of action potentials. They are composed of a pore-forming alpha subunit and are modulated by at least two of four distinct beta subunits (beta1-4). Recent studies have implicated a role for the intracellular domain of beta subunits in modulating Na channel gating and trafficking. In beta3, the intracellular domain contains a serine residue at position 161 that is replaced by an alanine in beta1. In this study, we have probed the functional importance of beta3S161 for modulating Na channel gating. Wild-type beta3 and point mutations beta3S161A or beta3S161E were individually co-expressed in HEK 293 cells stably expressing human Na(v)1.2. WTbeta3 expression increased Na current density, shifted steady-state inactivation in a depolarized direction, and accelerated the kinetics of recovery from inactivation of the Na current. Analogous effects were observed with beta3S161E co-expression. In contrast, beta3S161A abolished the shifts in steady state inactivation and recovery from inactivation of the Na current, but did increase Na current density. Immunocytochemistry and Western blot experiments demonstrate membrane expression of WTbeta3, beta3S161E, and beta3S161A, suggesting that the differences in Na channel gating were not due to disruptions in beta subunit trafficking. These studies suggest that modification of beta3S161 may be important in modulating Na-channel gating. PMID- 19806361 TI - Transgenic mice expressing nitroreductase gene under the control of the podocin promoter: a new murine model of inductible glomerular injury. AB - The present work identifies a new mouse model of inductible acute glomerular injury leading to focal segmental glomerulonephritis. We take advantage of the suicide gene/prodrug nitroreductase/CB1954 combination, in which nitroreductase converts CB1954, a monofunctional alkylating agent, into its toxic form. We generate two lines of transgenic mice in which the nitroreductase gene was placed under the control of the podocyte-specific gene podocin. The functional analysis of transgenic mice lines showed that CB1954 treatment induced a severe but transitory proteinuria. Sequential histopathological analysis was performed on serial kidney biopsies. Injured glomeruli showed acute lesions with early podocyte vacuolization and detachment, podocyte apoptosis, and cellular proliferation leading to a marked hypercellularity of the urinary space that was associated with collapsing of the glomerular tuft. After 1 month, progressive scarring lead to focal segmental glomerulosclerosis with fibrous capsular adhesion, hyalinosis, and podocytosis associated with interstitial fibrosis. The phenotype of podocytes was changed exhibiting dedifferentiation characterized by the loss of podocyte specific proteins/transcription factor and the expression of injury markers. Bowman's capsule cells were also involved in the cellular changes in a manner suggesting epithelial to mesenchymal transition. This model of podocyte injury in transgenic mice provides new insights into the cellular mechanisms of podocytopathies and their progression to scarring. PMID- 19806360 TI - Acid-evoked Ca2+ signalling in rat sensory neurones: effects of anoxia and aglycaemia. AB - Ischaemia excites sensory neurones (generating pain) and promotes calcitonin gene related peptide release from nerve endings. Acidosis is thought to play a key role in mediating excitation via the activation of proton-sensitive cation channels. In this study, we investigated the effects of acidosis upon Ca2+ signalling in sensory neurones from rat dorsal root ganglia. Both hypercapnic (pHo 6.8) and metabolic-hypercapnic (pHo 6.2) acidosis caused a biphasic increase in cytosolic calcium concentration ([Ca2+] i ). This comprised a brief Ca2+ transient (half-time approximately 30 s) caused by Ca2+ influx followed by a sustained rise in [Ca2+] i due to Ca2+ release from caffeine and cyclopiazonic acid-sensitive internal stores. Acid-evoked Ca2+ influx was unaffected by voltage gated Ca2+-channel inhibition with nickel and acid sensing ion channel (ASIC) inhibition with amiloride but was blocked by inhibition of transient receptor potential vanilloid receptors (TRPV1) with (E)-3-(4-t-butylphenyl)-N-(2,3 dihydrobenzo[b][1,4] dioxin-6-yl)acrylamide (AMG 9810; 1 microM) and N-(4 tertiarybutylphenyl)-4-(3-cholorphyridin-2-yl) tetrahydropryazine-1(2H)-carbox amide (BCTC; 1 microM). Combining acidosis with anoxia and aglycaemia increased the amplitude of both phases of Ca2+ elevation and prolonged the Ca2+ transient. The Ca2+ transient evoked by combined acidosis, aglycaemia and anoxia was also substantially blocked by AMG 9810 and BCTC and, to a lesser extent, by amiloride. In summary, the principle mechanisms mediating increase in [Ca2+] i in response to acidosis are a brief Ca2+ influx through TRPV1 followed by sustained Ca2+ release from internal stores. These effects are potentiated by anoxia and aglycaemia, conditions also prevalent in ischaemia. The effects of anoxia and aglycaemia are suggested to be largely due to the inhibition of Ca2+-clearance mechanisms and possible increase in the role of ASICs. PMID- 19806363 TI - Usefulness of pneumococcal antigen detection in pleural effusion for the rapid diagnosis of infection by Streptococcus pneumoniae. AB - Pleural effusion is increasingly reported in children. Standard culture of blood or pleural fluid is frequently negative and molecular diagnosis by polymerase chain reaction is not available in all hospitals. The Binax NOW Streptococcus pneumoniae Antigen test (Binax, Portland, USA) is a rapid immunochromatographic test (ICT) for the detection of the C polysaccharide antigen. We evaluated the Binax NOW test on the pleural fluid of 73 children hospitalized with pleural effusion over a period of 4 years. In our sample, the sensitivity and specificity of ICT were high (88% and 71%, respectively), with a positive predictive value of 96%. Detection of the pneumococcal antigen in pleural fluid by ICT is easy and quick, and enables us to identify the pneumococcal origin of the effusion, thus, making the treatment of complicated pneumonia suitably and early. PMID- 19806362 TI - Nodular pattern of bone marrow infiltration: frequent finding in immunosuppression-related EBV-associated large B-cell lymphomas. AB - Different patterns of bone marrow (BM) infiltration by diffuse large B cell lymphomas (DLBCL) have been described. A pure nodular pattern is uncommon, and the pathologic features, as well as the clinical correlates of DLBCL manifesting this pattern in the BM have not been well characterized. We evaluated BM biopsies involved by large B cell lymphomas diagnosed at our institute over an 11-year period to assess the morphology, phenotype, cytogenetic abnormalities, and clinical features of cases associated with a nodular pattern. A distinct nodular pattern of BM involvement was noted in 14 out of 55 (25%) cases. Although both EBV+ and EBV- DLBCL with this pattern were identified, a pure nodular pattern was significantly more common in EBV+ DLBCL compared to EBV- DLBCL (8/9, 89% versus 6/46, 13%; P = 0.00002). The majority of EBV+ DLBCL associated with a nodular pattern had distinctive morphologic features (polymorphic cellular infiltrate and pleomorphic cytology), and CD30 expression was more commonly observed in this group (P = 0.0163). All EBV+ DLBCL and two out of six (33%) EBV- DLBCL had nongerminal center phenotypes. No recurrent cytogenetic abnormalities were detected in either group. Importantly, all EBV+ DLBCL occurred in individuals with immune dysfunction (organ transplant recipients, HIV infection) or in those >50 years of age. Our study indicates a much higher predilection for EBV+ DLBCL to involve the marrow in a nodular pattern compared to EBV- cases and highlights similarities in the morphologic pattern of BM involvement by previously recognized subsets of immunodeficiency-related EBV + lymphomas and the newer entity of "EBV+ DLBCL of the elderly." PMID- 19806364 TI - Molecular identification and phylogenetic relationships of trichomonad isolates of galliform birds inferred from nuclear small subunit rRNA gene sequences. AB - Histomonas meleagridis is the etiological agent of histomonosis or blackhead disease. Recently, genotyping, based on polymerase chain reaction and sequencing of internal transcribed spacer-1 sequences was applied to various isolates originating from fowl. Three genotypes were described: types I and II isolates were associated with clinical disease and probably derived from H. meleagridis, whereas, type III isolates were not disease-associated and likely corresponded to Parahistomonas wenrichi according to morphological observations. However, this latter species has never been characterized at the molecular level and its phylogenetic relationships with other parabasalids remained hypothetical. To confirm the identification of these isolates, small subunit rRNA gene sequences were obtained from representatives of types I, II, and III and analyzed in a broad phylogeny including 64 other parabasalid sequences. From our phylogenetic trees, we confirmed that types I and II isolates were closely related, if not identical, to H. meleagridis, while type III isolates represented P. wenrichi. Both species clustered together with high support. This grouping suggested that speciation leading to these two species inhabiting the same hosts and ecological niche occurred recently in birds. In addition, speciation was likely followed by loss of pathogenicity in P. wenrichi. PMID- 19806365 TI - Borna disease virus infection alters synaptic input of neurons in rat dentate gyrus. AB - Granule cells are major targets of entorhinal afferents terminating in a laminar fashion in the outer molecular layer of the dentate gyrus. Since Borna disease virus (BDV) infection of newborn rats causes a progressive loss of granule cells in the dentate gyrus, entorhinal fibres become disjoined from their main targets. We have investigated the extent to which entorhinal axons react to this loss of granule cells. Unexpectedly, anterograde DiI tracing has shown a prominent layered termination of the entorhinal projection, despite an almost complete loss of granule cells at 9 weeks after infection. Combined light- and electron microscopic analysis of dendrites at the outer molecular layer of the dentate gyrus at 6 and 9 weeks post-infection has revealed a transient increase in the synaptic density of calbindin-positive granule cells and parvalbuminergic neurons after 6 weeks. In contrast, synaptic density reaches values similar to those of uninfected controls 9 weeks post-infection. These findings indicate that, after BDV infection, synaptic reorganization processes occur at peripheral dendrites of the remaining granule cells and parvalbuminergic neurons, including the unexpected persistence of entorhinal axons in the absence of their main targets. PMID- 19806366 TI - Implications of extreme sexual size dimorphism for thermoregulation in a freshwater turtle. AB - Sexual size dimorphism (SSD) is a common phenomenon in animals. In many species females are substantially larger than males. Because body size plays a central role in modulating the body temperature (T (b)) of ectotherms, intersexual differences in body size may lead to important intersexual differences in thermoregulation. In addition, because SSD is realized by differences in growth rate and because growth rate is strongly temperature dependent in ectotherms, a conflict between male reproductive behaviour and thermoregulation may affect the expression of SSD. In this study, we investigated the thermal implications of SSD in a reptile exhibiting spectacular female-biased SSD: the northern map turtle (Graptemys geographica). Over three seasons, we collected >150,000 measurements of T (b) in free-ranging adult and juvenile northern map turtles using surgically implanted miniature temperature loggers. Northern map turtles exhibited seasonal patterns of thermoregulation typical of reptiles in northern latitudes, but we found that large adult females experienced a lower daily maximum T (b) and a narrower daily range of T (b) than adult males and small juvenile females. In addition, despite more time spent basking, large adult females were not able to thermoregulate as accurately as small turtles. Our findings strongly suggest that body size limits the ability to thermoregulate accurately in large females. By comparing thermoregulatory patterns between adult males and juvenile females of similar body size, we found no evidence that male reproductive behaviours are an impediment to thermoregulation. We also quantified the thermal significance of basking behaviour. We found, contrary to previous findings, that aerial basking allows northern map turtles to raise their T (b) substantially above water temperature, indicating that basking behaviour likely plays an important role in thermoregulation. PMID- 19806367 TI - Inhibition of lipoxygenase affects induction of both direct and indirect plant defences against herbivorous insects. AB - Herbivore-induced plant defences influence the behaviour of insects associated with the plant. For biting-chewing herbivores the octadecanoid signal transduction pathway has been suggested to play a key role in induced plant defence. To test this hypothesis in our plant-herbivore-parasitoid tritrophic system, we used phenidone, an inhibitor of the enzyme lipoxygenase (LOX), that catalyses the initial step in the octadecanoid pathway. Phenidone treatment of Brussels sprouts plants reduced the accumulation of internal signalling compounds in the octadecanoid pathway downstream of the step catalysed by LOX, i.e. 12-oxo phytodienoic acid (OPDA) and jasmonic acid. The attraction of Cotesia glomerata parasitoids to host-infested plants was significantly reduced by phenidone treatment. The three herbivores investigated, i.e. the specialists Plutella xylostella, Pieris brassicae and Pieris rapae, showed different oviposition preferences for intact and infested plants, and for two species their preference for either intact or infested plants was shown to be LOX dependent. Our results show that phenidone inhibits the LOX-dependent defence response of the plant and that this inhibition can influence the behaviour of members of the associated insect community. PMID- 19806369 TI - Editorial--Nutrition as supportive care in the cancer experience. PMID- 19806368 TI - Dent's disease manifesting as focal glomerulosclerosis: Is it the tip of the iceberg? AB - Dent's disease is an X-linked proximal tubulopathy. It often manifests in childhood with symptoms of Fanconi syndrome and low-molecular-weight proteinuria. We describe four boys from three unrelated families whose only presenting symptoms of Dent's disease were nephrotic-range proteinuria and histological findings of focal segmental and/or global glomerulosclerosis. In all families, a causal mutation in the CLCN5 gene, encoding a voltage-gated chloride transporter and chloride-proton exchanger, was identified. All three mutations are pathogenic: two are novel (p.Asp727fs and p.Trp122X), and one is a recurrent mutation, p.R648X. Given the atypical phenotype of these patients with Dent's disease, it is possible that this clinical entity is markedly underdiagnosed and that our report represents only the tip of the iceberg. The diagnosis of Dent's disease should be considered in all patients with nephrotic-range proteinuria without hypoalbuminemia or edema. Establishing the diagnosis of Dent's disease will prevent the administration of unnecessary immunosuppressive medications with their undesirable side effects. PMID- 19806370 TI - Attitudes toward end-of-life situations other than euthanasia and assisted suicide among Portuguese oncologists. AB - BACKGROUND: The aim of this study was to determine the attitude of Portuguese oncologists toward end-of-life situations other than euthanasia and assisted suicide. METHODS: This study used a survey of 450 Portuguese oncologists by postal means and personal contact. MAIN RESULTS: The response rate was 33% (143). Only 7.7% doctors would give lethal doses of drugs to someone with an incurable, advanced, and progressive disease that is unable to make decisions, at the request of a family member or other close person. However, 30 doctors (21.3%) would prefer, in the event they were in such a situation, that the drugs be given them at their request. None of the 12.4% who have received such requests admitted to committing any of those acts. Almost 70% of the doctors would withdraw life support measures at the patient's request if the same had an incurable, advanced, and progressive disease, and a further 14% would do it in certain circumstances, but only 41% would withdraw measures such as nutrition and hydration. Fewer doctors would withdraw such measures including nutrition and hydration at a family member's request or on their own initiative. Religion has a major influence on the doctors' opinion. Most doctors (96.5%) agreed with the administration of drugs for symptom control even foreseeing that they could shorten life. CONCLUSION: Most Portuguese oncologists respect patients' autonomy, favoring the withdrawal of life support treatment at the patients' request when appropriate and much less do so on the request of others or by their own initiative. They appropriately do not confuse those practices with symptom relief even when hastened death can be envisaged. PMID- 19806371 TI - Assessment of 3-D nasal airway morphology in Southeast Asian adults with obstructive sleep apnea using acoustic rhinometry. AB - The aim of the present study is to investigate nasal airway morphology in Asian adults with and without obstructive sleep apnea (OSA) using acoustic rhinometry (AR), principal components analysis (PCA), and 3-D finite-element analysis (FEA). One hundred eight adult Malays aged 18-65 years (mean +/- SD, 33.2 +/- 13.31) underwent clinical examination and limited channel polysomnography, providing 54 patients with OSA and 54 non-OSA controls. The mean minimal cross section area 1 (MCA1) and the mean minimal cross sectional area 2 (MCA2) were obtained from AR for all subjects and subjected to t tests. The OSA and control nasal airways were reconstructed in 3-D and subjected to PCA and FEA. The mean MCA1 and MCA2 using AR were found to be significantly smaller in the OSA group than in the control group (p < 0.001). Comparing the 3-D OSA and control nasal airways using PCA, the first two eigenvalues accounted for 94% of the total shape change, and statistical differences were found (p < 0.05). Similarly, comparing the nasal airways using FEA, the 3-D mean OSA nasal airway was significantly narrower in the OSA group compared to the control group. Specifically, decreases in size of approx. 10-22% were found in the nasal valve/head of inferior turbinate area. In conclusion, differences in nasal airway morphology are present when comparing patients with OSA to controls. These differences need to be recognized as they can improve our understanding of the etiological basis of obstructive sleep apnea and facilitate its subsequent management. PMID- 19806372 TI - Treatment of temporomandibular joint ankylosis: gap and interpositional arthroplasty with temporalis muscle flap. AB - PURPOSE: The purpose of this study was to examine the effectiveness of gap and interpositional arthroplasty with temporalis muscle flap in the treatment of the temporomandibular joint (TMJ) ankylosis. MATERIALS AND METHODS: This retrospective study was evaluated on 34 patients who were treated from February 2003 to July 2007. Diagnosis of TMJ ankylosis was based upon the clinical examination and plain radiography and CT scan. Eight patients (23.5%) had bilateral TMJ ankylosis, and four patients had reankylosis (that had been operated in another center). Trauma was the most etiological factor (88.2%) and osteochondroma of the condyle in one patient caused ankylosis. RESULTS: Means of maximum mouth opening before and after operation were 5.00 (SD) 3.45 mm and 32.85 (SD) 5.51 mm, respectively. Reankylosis in two patients (5.9%) and facial nerve dysfunction in 12 surgery sides (28.5%) were noted. CONCLUSION: The findings of this study suggest that the gap arthroplasty with temporalis muscle flap as interpositional graft is an effective method in the treatment of TMJ ankylosis. The osteoarthrectomy of the callus to create at least 10 mm gap and enough bulk of temporalis muscle flap as interpositional graft followed by at least a 6-month physiotherapy which play an important role in prevention of reankylosis. PMID- 19806373 TI - Revisiting the phenotype associated with FOXG1 mutations: two novel cases of congenital Rett variant. AB - The Forkhead box G1 (FOXG1) is a transcription factor that is critical for forebrain development, where it promotes progenitor proliferation and suppresses premature neurogenesis. Recently, the FOXG1 gene was implicated in the molecular aetiology of the congenital variant of Rett syndrome. So far, 15 FOXG1 molecular alterations, including only eight point mutations, have been reported. We screened the FOXG1 gene in a cohort of 206 MECP2 and CDKL5 mutation negative patients (136 females and 70 males) with severe encephalopathy and microcephaly. The screening was negative in all males, but two de novo mutations (c.1248C>G, p.Y416X and c.460_461dupG, p.E154GfsX300) were identified in two unrelated girls. Both patients showed neurological symptoms from the neonatal period with poor reactivity, hypotonia, and severe microcephaly. During the first year of life, both patients had feeding difficulties and made slow developmental progress. At 5 years old, the girls were significantly neurologically impaired with gross hypotonia, no language, convergent strabismus, and no voluntary hand use. Moreover, they presented a combination of jerky movements, hand-mouthing, and hand-washing stereotypies. Hence, FOXG1 mutation patients demonstrate severe encephalopathy compatible with the congenital variant, as well as additional features such as absent eye contact, inconsolable crying during the perinatal period, and delayed myelination with thin to hypoplastic corpus callosum. Although the overall frequency of mutations in FOXG1 in females with severe mental retardation and microcephaly appears to be low (1.5%), our findings suggest the requirement to investigate both point mutations and gene dosage in the FOXG1 gene in patients with severe encephalopathy with microcephaly and some Rett-like features. PMID- 19806374 TI - Molecular characteristics of pharyngeal and invasive emm3 Streptococcus pyogenes strains from Norway, 1988-2003. AB - A major virulence factor of group A streptococci (GAS) is the M protein. Strains with the M3 type are more often associated with necrotizing fasciitis (NF) and streptococcal toxic shock syndrome, and have a higher case fatality rate than strains of other M types. To better understand the epidemiology of M3 GAS strains in Norway, we analyzed 59 invasive and 69 pharyngeal isolates with respect to prophage content, allelic variation in emm3, mtsR encoding the metal transporter of Streptococcus repressor (mtsR), and sclB coding for streptococcal collagen like protein B. The Norwegian emm3 strains were very homogeneous, mainly harboring the emm allele 3.1 and prophage profile PhiG3.01. Other prophage profiles were transient. The mutation in mtsR known to truncate the protein and result in decreased capacity to cause NF was not found in our isolates. The sclB gene usually harbored five or eight contiguous repeats of a CAAAA pentanucleotide sequence and a highly modular and variable collagen structural motif (CSM) region with 9 and 12 amino acid M3-specific conserved motif repeats distributed across the entire CSM region. Strains with 5 CAAAA repeats emerged in 1993 and these strains were associated with the increase in invasive M3 cases in the period 1993 2003. PMID- 19806375 TI - Influence of cultivation conditions on the production of a thermostable extracellular lipase from Amycolatopsis mediterranei DSM 43304. AB - Among several lipase-producing actinomycete strains screened, Amycolatopsis mediterranei DSM 43304 was found to produce a thermostable, extracellular lipase. Culture conditions and nutrient source modification studies involving carbon sources, nitrogen sources, incubation temperature and medium pH were carried out. Lipase activity of 1.37 +/- 0.103 IU/ml of culture medium was obtained in 96 h at 28 degrees C and pH 7.5 using linseed oil and fructose as carbon sources and a combination of phytone peptone and yeast extract (5:1) as nitrogen sources. Under optimal culture conditions, the lipase activity was enhanced 12-fold with a twofold increase in lipase specific activity. The lipase showed maximum activity at 60 degrees C and pH 8.0. The enzyme was stable between pH 5.0 and 9.0 and temperatures up to 60 degrees C. Lipase activity was significantly enhanced by Fe(3+) and strongly inhibited by Hg(2+). Li(+), Mg(2+) and PMSF significantly reduced lipase activity, whereas other metal ions and effectors had no significant effect at 0.01 M concentration. A. mediterranei DSM 43304 lipase exhibited remarkable stability in the presence of a wide range of organic solvents at 25% (v/v) concentration for 24 h. These features render this novel lipase attractive for potential biotechnological applications in organic synthesis reactions. PMID- 19806376 TI - On the relationship between ozone and its precursors in the Pearl River Delta: application of an observation-based model (OBM). AB - BACKGROUND, AIM, AND SCOPE: Photochemical smog, characterized by high concentrations of O(3) and fine particles, is of great concern in the urban areas, in particular megacities and city clusters like the Pearl River Delta. MATERIALS, METHODS, AND RESULTS: Ambient ozone (O(3)) and its precursors were simultaneously measured at two sites in the Pearl River Delta, namely, Wan Qing Sha (WQS) in Guangzhou and Tung Chung (TC) in Hong Kong, from 23 October to 01 December 2007 in order to explore their potential relationship. Eight high O(3) episode days were identified at WQS and two at TC during the sampling campaign, indicating a more serious O(3) pollution in Guangzhou than in Hong Kong. An observation-based model was employed to determine the ozone-precursor relationship. At both sites, O(3) production was found to be volatile organic compound (VOC)-limited, which is consistent with previous observations. Anthropogenic hydrocarbons played a key role in O(3) production, while reducing nitric oxide emissions aided the buildup of O(3) concentrations. Among VOC species, the summed relative incremental reactivity (RIR) of the top 12 compounds accounted for 89% and 85% of the total RIR at WQS and TC, respectively, indicating that local photochemical O(3) formation can be mainly attributed to a small number of VOC species. DISCUSSION AND CONCLUSIONS: A large increment in both simulated HO(2) and O(3) concentrations was achieved with additional input of hourly carbonyl data. This suggested that apart from hydrocarbons, carbonyls might significantly contribute to the O(3) production in the Pearl River Delta. PMID- 19806377 TI - Characteristics of exhaust gas, liquid products, and residues of printed circuit boards using the pyrolysis process. AB - INTRODUCTION: The pyrolytic method was employed to recycle metals and brominated compounds blended into printed circuit boards (PCBs). METHODS: PCBs were crushed into pieces 4.0-4.8 mm in size, and the crushed pieces were pyrolyzed at temperatures ranging from 200 to 500 degrees C. The compositions of pyrolytic residues, liquid products, and exhaust were analyzed by inductively coupled plasma atomic emission spectrometer, inductively coupled plasma mass spectrometry, and gas chromatography-mass spectrometry. Pyrolytic exhaust was collected by an impinger system in an ice bath cooler to analyze the composition fraction of the liquid product, and uncondensable exhaust was collected for gas constituent analysis. RESULTS: Phenol, methyl-phenol, and bromo-phenol were attributed mainly to the liquid product. Metal content was low in the liquid product. In addition, CO, CO(2), CH(4), and H(2) were the major components of pyrolytic exhaust. CONCLUSIONS: Brominated and chlorinated compounds-i.e., dichloromethane, trans-1,2 dichloroethylene, cis-1,2 dichloroethylene, 1,1,1 trichloroethane, tetrachloromethane, bromophenol, and bromoform-could be high, up to the several parts per million (ppm) level. Low molecular weight volatile organic compounds (VOCs)-i.e., methanol, acetone, ethyl acetate, acrylonitrile, 1 butene, propene, propane, and n-butane-contributed a large fraction of VOCs. The concentrations of toluene, benzene, xylene, ethylbenzene, and styrene were in the ppm range. PMID- 19806378 TI - Cocoa butter and safflower oil elicit different effects on hepatic gene expression and lipid metabolism in rats. AB - The aim of this study was to compare the effects of cocoa butter and safflower oil on hepatic transcript profiles, lipid metabolism and insulin sensitivity in healthy rats. Cocoa butter-based high-fat feeding for 3 days did not affect plasma total triglyceride (TG) levels or TG-rich VLDL particles or hepatic insulin sensitivity, but changes in hepatic gene expression were induced that might lead to increased lipid synthesis, lipotoxicity, inflammation and insulin resistance if maintained. Safflower oil increased hepatic beta-oxidation, was beneficial in terms of circulating TG-rich VLDL particles, but led to reduced hepatic insulin sensitivity. The effects of safflower oil on hepatic gene expression were partly overlapping with those exerted by cocoa butter, but fewer transcripts from anabolic pathways were altered. Increased hepatic cholesterol levels and increased expression of hepatic CYP7A1 and ABCG5 mRNA, important gene products in bile acid production and cholesterol excretion, were specific effects elicited by safflower oil only. Common effects on gene expression included increased levels of p8, DIG-1 IGFBP-1 and FGF21, and reduced levels of SCD-1 and SCD-2. This indicates that a lipid-induced program for hepatic lipid disposal and cell survival was induced by 3 days of high-fat feeding, independent on the lipid source. Based on the results, we speculate that hepatic TG infiltration leads to reduced expression of SCD-1, which might mediate either neutral, beneficial or unfavorable effects on hepatic metabolism upon high-fat feeding, depending on which fatty acids were provided by the diet. PMID- 19806379 TI - Imaging on pelvic pyomyositis in children related to pathogenesis. AB - PURPOSE: Pelvic pyomyositis in children is a rare infectious condition, although it is increasingly reported in temperate climates. Often considered a primary disease, new diagnostic methods are able to identify additional foci of infection. The purpose of this study is to review our patients and to analyze the imaging studies to determine its pathogenesis. METHODS: A retrospective study of the clinical charts and imaging records of 11 patients was made, noting the number and location of muscles involved, as well as bone and joint involvement. RESULTS: Besides the classical form of pelvic pyomyositis, i.e., iliopsoas pyomyositis, other muscular groups were frequently affected, often with multiple involvement. Bone involvement is also frequent. Magnetic resonance imaging (MRI) gives the most useful information. CONCLUSION: MRI is the diagnostic procedure of choice for diagnosing pelvic pyomyositis in children. It may also have an elucidating role in the debated pathogenesis of this condition. In most of the cases, pelvic pyomyositis in children could be secondary. PMID- 19806381 TI - [Visual self-assessment with the ACTO test during follow-up of AMD patients after intravitreal injections]. AB - INTRODUCTION: Monthly controls are necessary after injections of vascular endothelial growth factor (VEGF) to enable timely recognition of a renewed decrease in vision. However, these monthly control intervals are not realistic for many older patients with age-related macular degeneration (AMD), and outpatient clinics often reach their logistical limits because of inadequate funding for the additional medical work. Against this background, we conceived the ACTO self-test as a novel screening method for patients to discover unnoticed visual changes outside the routinely scheduled ophthalmic examination. MATERIAL AND METHODS: The paper version of the ACTO self-test consists of a reading chart gradated in decimal steps as well as six questions regarding the quality of the Amsler grid. The patient uses a self-screening test to examine each eye separately and then transfers the results to a table. Along with the self examination, the Action Eyesight Service Center is available to the patient by phone to motivate the patient and schedule a new appointment if improved test results occur. If decreased values or suspected decreased visual function occur, these are verified immediately by the referring physician. RESULTS: There is a good correlation between the steps of the ACTO self-test and standard visual acuity at 4 m (r(2)=0.9). Altogether, 1,444 patients were followed by phone and 745 participated in repeated regular audits, for a total of 3,003 phone contacts. The treating physician was informed about decreased visual acuity or increased Amsler distortion in the ACTO test in 137 cases, and immediate verification was done. We had 699 dropouts; the reasons were decreased visual acuity below the limits of the ACTO test in 39%, additional monthly examinations by the local ophthalmologist in 29%, and a desire for no more telephone follow-up (despite initial written consent) in 32%. Decreased vision when reading or an increased score on the Amsler test within the ACTO screening test was observed in 18% of AMD patients during the maintenance phase. CONCLUSION: Monthly screening by the ACTO screening test in combination with phone audits offers a new way to test visual acuity, with the Amsler score helping to detect changes in visual function. For patients with visual changes, confirmation by the ophthalmic physician can be achieved in time. Self-assessment cannot replace qualified ophthalmologic examination, but monthly self-controls enhance safety, reduce the number of physician contacts, and improve the detection of visual changes, with the option of immediate ophthalmic retreatment. PMID- 19806380 TI - Chemopreventive potential of curcumin in prostate cancer. AB - The long latency and high incidence of prostate carcinogenesis provides the opportunity to intervene with chemoprevention in order to prevent or eradicate prostate malignancies. We present here an overview of the chemopreventive potential of curcumin (diferuloylmethane), a well-known natural compound that exhibits therapeutic promise for prostate cancer. In fact, it interferes with prostate cancer proliferation and metastasis development through the down regulation of androgen receptor and epidermal growth factor receptor, but also through the induction of cell cycle arrest. It regulates the inflammatory response through the inhibition of pro-inflammatory mediators and the NF-kappaB signaling pathway. These results are consistent with this compound's ability to up-induce pro-apoptotic proteins and to down-regulate the anti-apoptotic counterparts. Alone or in combination with TRAIL-mediated immunotherapy or radiotherapy, curcumin is also reported to be a good inducer of prostate cancer cell death by apoptosis. Curcumin appears thus as a non-toxic alternative for prostate cancer prevention, treatment or co-treatment. PMID- 19806382 TI - [Development of a glaucoma microstent with drainage into the suprachoroidal space: fluid mechanical model approach]. AB - BACKGROUND: The purpose of this study was to develop a microstent with valve function, which normalizes the intraocular pressure (IOP) and drains into the suprachoroidal space. In comparison to the subconjunctival space the suprachoroidal space is attributed with less fibroblast colonization and activity. METHODS: Different glaucoma drainage devices were idealized as tubes and the flow rates were calculated according to Hagen-Poiseuille. The dimensions of the ideal glaucoma implant were modified with respect to an aqueous humor production of 2 microl/min and the different outflow pathways. Specific components of glaucoma drainage devices at the inlet and outlet were not included. RESULTS: The volume flow calculation of the tested glaucoma implants showed that the dimensions of all lumina were too large to prevent postoperative hypotension. A maximum inner tube diameter of 53 microm was calculated for drainage into the suprachoroidal space based on an intra-ocular pressure (IOP) of 20 mmHg. CONCLUSION: The glaucoma microstent has to guarantee an aqueous humor flow for physiological IOP. An increase of IOP has to be regulated to physiological pressure conditions by the microvalve. PMID- 19806383 TI - [Squamous cell carcinoma of the "pure" cornea]. AB - Squamous cell carcinoma of the cornea without involvement of the conjunctiva and limbus is a very rare condition. We report on a 68-year-old female patient. Lamellar keratectomy was performed combined with brachytherapy with strontium-90 for a total dose of 120 Gy (single daily dose: 10 Gy). Two months after treatment, a regression of neovascularization was noted. During 3.5 years of follow-up, no rubeosis of the iris, secondary glaucoma, or evidence of tumor recurrence was seen. There were no regional or distant metastases. PMID- 19806384 TI - Elastin degradation accelerates phosphate-induced mineralization of vascular smooth muscle cells. AB - Medial layer vascular calcification is common in patients with end-stage kidney disease. Inorganic phosphate has been shown to accelerate the transformation of vascular smooth muscle cells (VSMCs) into osteoblast-like cells, which is thought to be a major process of medial layer calcification. Although elastin degradation is associated with medial layer calcification, the linkage between elastin degradation and the transformation of VSMCs remains to be clarified. We investigated the involvement of elastin degradation in the transformation of VSMCs. Rat VSMCs were isolated and cultured with a normal- (NP, 1.0 mM) or high- (HP, 2.5 mM) phosphate medium. An elastin-derived peptide, alpha-elastin (500 microg/ml), was also added to the normal- (NP + E) or high- (HP + E) phosphate medium. After a culture period of 2 weeks, von Kossa staining revealed mineralization in the HP group, which was accelerated by alpha-elastin, whereas alpha-elastin did not affect the mineralization at a normal phosphate concentration. The gene expression of osteoblastic differentiation factors, i.e., Runx2 or osteocalcin (OC), in VSMCs was significantly increased in the HP (Runx2 P < 0.05, OC P < 0.05) and HP + E (OC P < 0.05) groups compared with the NP and NP + E groups. Both gene and protein expressions of tissue-nonspecific alkaline phosphatase (TNAP) were significantly increased in the HP group compared with the NP and NP + E groups (P < 0.01, respectively). This increment was augmented in the HP + E group (P < 0.01). These results suggest that elastin degradation would accelerate or stabilize the process of VSMC transformation, which is induced by high phosphate through the upregulation of TNAP. PMID- 19806385 TI - Optical coherence tomography phase measurement of transient changes in squid giant axons during activity. AB - Noncontact optical measurements reveal that transient changes in squid giant axons are associated with action potential propagation and altered under different environmental (i.e., temperature) and physiological (i.e., ionic concentrations) conditions. Using a spectral-domain optical coherence tomography system, which produces real-time cross-sectional images of the axon in a nerve chamber, axonal surfaces along a depth profile are monitored. Differential phase analyses show transient changes around the membrane on a millisecond timescale, and the response is coincident with the arrival of the action potential at the optical measurement area. Cooling the axon slows the electrical and optical responses and increases the magnitude of the transient signals. Increasing the NaCl concentration bathing the axon, whose diameter is decreased in the hypertonic solution, results in significantly larger transient signals during action potential propagation. While monophasic and biphasic behaviors are observed, biphasic behavior dominates the results. The initial phase detected was constant for a single location but alternated for different locations; therefore, these transient signals acquired around the membrane appear to have local characteristics. PMID- 19806386 TI - Membrane porters of ATP-binding cassette transport systems are polyphyletic. AB - The ATP-binding cassette (ABC) superfamily consists of both importers and exporters. These transporters have, by tradition, been classified according to the ATP hydrolyzing constituents, which are monophyletic. The evolutionary origins of the transmembrane porter proteins/domains are not known. Using five distinct computer programs, we here provide convincing statistical data suggesting that the transmembrane domains of ABC exporters are polyphyletic, having arisen at least three times independently. ABC1 porters arose by intragenic triplication of a primordial two-transmembrane segment (TMS)-encoding genetic element, yielding six TMS proteins. ABC2 porters arose by intragenic duplication of a dissimilar primordial three-TMS-encoding genetic element, yielding a distinctive protein family, nonhomologous to the ABC1 proteins. ABC3 porters arose by duplication of a primordial four-TMS-encoding genetic element, yielding either eight- or 10-TMS proteins. We assign each of 48 of the 50 currently recognized families of ABC exporters to one of the three evolutionarily distinct ABC types. Currently available high-resolution structural data for ABC porters are fully consistent with our findings. These results provide guides for future structural and mechanistic studies of these important transport systems. PMID- 19806387 TI - The origin of life: chemical evolution of a metabolic system in a mineral honeycomb? AB - For the RNA-world hypothesis to be ecologically feasible, selection mechanisms acting on replicator communities need to be invoked and the corresponding scenarios of molecular evolution specified. Complementing our previous models of chemical evolution on mineral surfaces, in which selection was the consequence of the limited mobility of macromolecules attached to the surface, here we offer an alternative realization of prebiotic group-level selection: the physical encapsulation of local replicator communities into the pores of the mineral substrate. Based on cellular automaton simulations we argue that the effect of group selection in a mineral honeycomb could have been efficient enough to keep prebiotic ribozymes of different specificities and replication rates coexistent, and their metabolic cooperation protected from extensive molecular parasitism. We suggest that mutants of the mild parasites persistent in the metabolic system can acquire useful functions such as replicase activity or the production of membrane components, thus opening the way for the evolution of the first autonomous protocells on Earth. PMID- 19806389 TI - Ostial stenosis of an anomalous left coronary artery from the pulmonary artery in a teenager. PMID- 19806390 TI - Evaluating safety, effectiveness, and user satisfaction of home international normalized ratio monitoring service: experience from a tertiary pediatric cardiology unit in the United Kingdom. AB - Home international normalized ratio (INR) monitors are being increasingly used for monitoring the adequacy of anticoagulation in children on long-term warfarin. Their safety and effectiveness in the home setting has not been fully established. The purpose of this study was to explore the safety and effectiveness of home INR monitoring service in managing anticoagulation in children with congenital heart disease. At the Pediatric Cardiology Regional Referral Center, we studied records of 41 children on warfarin and assessed the control of anticoagulation and frequency of adverse reactions over a 1-year period. User satisfaction was assessed by means of telephone interviews. The study found that the therapeutic INR range was maintained for over 57% of the total study period. There were no reported instances of major or minor hemorrhages. On six occasions, subtherapeutic INR levels necessitated hospitalization for heparinization. Mitral valve replacement was significantly associated with poor control (p < 0.05). User satisfaction was high for all aspects of the service evaluated. The hospital-assisted home INR monitoring service is a safe and effective means of managing anticoagulation in children. In addition, it ensures good compliance and a high level of user satisfaction. PMID- 19806391 TI - A new postoperative otoplasty dressing technique using cyanoacrylate tissue adhesives. AB - There are many techniques for cosmetic surgery of the ears and also many different procedures for postoperative treatment. The postoperative dressing is described as important for a successful outcome. We present our method of postoperative dressing in the form of liquid bonding. Cyanoacrylate tissue adhesives as liquid bonding agents are used for fixation of the pinna at the mastoid area. After 10-14 days the bonding can be easily removed. No huge dressings, tapes, or plasters are necessary. The patients are satisfied with the light dressing; they do not feel ashamed to appear in public. We have found this dressing technique to be simple and economical, especially because of the use of the bonding for skin closure before. It can be used after otoplasty with an anterior or a posterior approach. PMID- 19806393 TI - The birth of pancreatic surgery: a tribute to Friedrich Wilhelm Wandesleben. AB - Friedrich Wilhelm Wandesleben (1800-1868), a small-town German physician, performed surgical drainage of a traumatic pancreatic pseudocyst in November 1841. This operation should be acknowledged as the world's first reported operation on the human pancreas. The surgeon, the operation, and its implications for future pancreatic operations are the subjects of this historical review. PMID- 19806396 TI - Archaeal diversity in the haloalkaline Lake Elmenteita in Kenya. AB - A non-culture approach was used to study the archaeal diversity in Lake Elmenteita, Kenya. Five different sampling points were selected randomly within the lake. Wet sediments and water samples were collected from each sampling point. In addition, dry mud cake was collected from three points where the lake had dried. DNA was extracted from these samples and the 16S rRNA genes were amplified using primers described to be Domain-specific for Archaea. Eleven clone libraries were constructed using PCR-amplified 16S rRNA genes. A total of 1,399 clones were picked and analysed via ARDRA. 170 ARDRA patterns were unique and the respective clones were selected for sequencing. 149 clones gave analysable sequences. BLAST analysis showed that 49 belong to the Domain Archaea while the others were either chimera or affiliated to eukaryotic taxa. Comparative sequence analysis of archaeal clones affiliated them to a wide range of genera. The order Halobacteriales was represented by members of the genera Natronococcus, Halovivax, Halobiforma, Halorubrum, and Halalkalicoccus. The highest percentage (46%) of the clones, however, belonged to uncultured members of the Domain Archaea in the order Halobacteriales. The results show that the archaeal diversity in the lake could be higher than previously reported. PMID- 19806397 TI - Type IV pili of Acidithiobacillus ferrooxidans are necessary for sliding, twitching motility, and adherence. AB - We used conventional methods to investigate the mechanism by which Acidithiobacillus ferrooxidans colonizes a solid surface by assessing pili mediated sliding, twitching motility, and adherence. A. ferrooxidans slided to form circular oxidized zones around each colony. This suggested that slide motility occurs through pili or flagella, though A. ferrooxidans strains ATCC 19859 and ATCC 23270 lack flagella. The results of reverse transcription-PCR demonstrated that the putative major pili gene of A. ferrooxidans strains ATCC 19859, ATCC 23270, and BY3 genes were transcribed. Culture of A. ferrooxidans between silicone gel and glass led to the production of type IV pili and the formation of rough twitching motility zones. When the bacteria were grown on lean ore cubes, pyrite was colonized readily by A. ferrooxidans and there is a correlation between pilus expression and strong attachment. However, non-pili bacteria attached minimally to the mineral surface. The results show a correlation between these functions and pilus expression. PMID- 19806398 TI - Genetic basis of sex-specific resistance to neuro-oncogenesis in (BDIX x BDIV) F(2) rats. AB - The identification of cancer susceptibility- and resistance-mediating genes is an essential prerequisite for prevention and early diagnosis of malignant tumors. Model organisms are helpful to identify variant alleles involved in pathways affecting individual cancer risk. BDIX and BDIV rats of both sexes are highly susceptible and resistant, respectively, to the development of N-ethyl-N nitrosourea (ENU)-induced malignant peripheral nerve sheath tumors (MPNST), predominantly in the trigeminal nerves. Nevertheless, female (BDIV x BDIX) F(2) intercross rats have a lower MPNST incidence and a longer latency time than males. Six of seven autosomal gene loci (Mss1-Mss7) controlling genetic susceptibility and resistance in (BDIV x BDIX) F(2) hybrids exert allele- and sex specific effects on tumor incidence and/or latency time of variable strength. Homozygous BDIV alleles at Mss4 or Mss7 located on rat chromosomes 6 and 10, respectively, are sufficient to cause almost complete resistance to ENU-induced MPNST development in female F(2) rats regardless of the genotype of the other locus. Both loci display only weak effects on male cancer risk. Survival curves of ENU-treated F(2) females depleted of animals with homozygous BDIV alleles at Mss4 and Mss7 are not significantly different from those of males, suggesting that these loci account mainly for the excess tumor resistance observed in female F(2) rats. By haplotype analysis Mss4 and Mss7 could be narrowed down to 20 and 12 Mb, respectively, providing a basis for the positional identification of candidate genes. PMID- 19806399 TI - NHLBI training workshop report: the vanishing pediatric pulmonary investigator and recommendations for recovery. AB - The adequacy of the pipeline of advanced pulmonary fellows to supply appropriately trained and committed researchers to enter academic careers was the major topic of a recently held National Heart Lung and Blood Institute NHLBI Workshop: Respiratory Medicine-Related Research Training for Adult and Pediatric Fellows. The special challenges and opportunities for the academic pediatric pulmonary trainee were discussed as part of this workshop and are presented as a companion article to the report by the full workshop. Surveys were conducted of pediatric chairs of academic departments and pediatric pulmonary training directors in the United States to examine the current status and opportunities for the pediatric pulmonary trainee. Strategies for recruitment and retention of talented young trainees and junior faculty are proposed. PMID- 19806401 TI - The lung allocation score and survival in lung transplant candidates with chronic obstructive pulmonary disease. AB - The Lung Allocation Score (LAS), devised to prioritize candidates awaiting lung transplantation (LTX), is calculated using the predicted duration of survival on the wait list while also considering the recipient's likelihood of post transplant survival. This score is generated based, in part, on the severity of the candidate's comorbid illnesses. The actual relationship between the LAS and survival is unknown. The current study was performed to evaluate the relationship between the LAS and both wait-list survival and post-transplant survival in candidates with COPD. The study was a retrospective analysis of 41 LTX candidates with chronic obstructive pulmonary disease (COPD) as well as a cohort of 17 candidates who survived to receive a graft. The study was conducted at a university hospital transplant center. Thirty-six of 41 candidates survived to transplant. The LAS of these survivors was 32.62 +/- 1.06 and was significantly lower than the score of 34.45 +/- 1.19 of the nonsurvivors (P < 0.01). The LAS also exhibited a negative association with survival to transplant (P < 0.05, beta = -1.39). A cohort of 17 LTX recipients was chosen for post-transplant analysis in which 13 survived at least 1 year. In this cohort the LAS did not exhibit significant association with 1-year post-transplant survival (P = 0.58, beta = 0.25). As might be anticipated by virtue of its calculation being based in part on the existence and severity of comorbid conditions, a lower LAS was associated with improved survival to transplantation in LTX candidates with COPD. However, the pretransplant calculation of the LAS was not associated with actual post transplant survival. PMID- 19806400 TI - Dexamethasone and cyclic AMP regulate sodium phosphate cotransporter (NaPi-IIb and Pit-1) mRNA and phosphate uptake in rat alveolar type II epithelial cells. AB - Alveolar epithelial type II (AT II) cells need phosphate (Pi) for surfactant synthesis. The Na-dependent (Na(d)) Pi transporters NaPi-IIb and Pit-1 are expressed in lung, but their expression, regulation, and function in AT II cells remain unclear. We studied NaPi-IIb and Pit-1 mRNA expression in cultured AT II cells isolated from adult rat lung, their regulation by agents known to enhance surfactant production, dexamethasone (dex) and dibutyryl cyclic AMP (cAMP), and the effects of dex and cAMP on Na(d) Pi uptake by this cell type. By Northern analysis, cultured AT II cells expressed both NaPi-IIb (4.8 and 4.0 kb) and Pit-1 (4.3 kb) mRNA. Treatment with 100 nmol/l dex for 24 h decreased the expression of both mRNAs (to 0.48 +/- 0.06 and 0.77 +/- 0.05, respectively, as compared to control), while 0.1 mmol/l cAMP stimulated NaPi-IIb (1.94 +/- 0.22) but not Pit-1 mRNA (0.90 +/- 0.05, compared to vehicle-treated cells). NaPi-IIb and Pit-1 proteins could not be identified by western analysis of plasma membrane preparations of cultured AT II cells. AT II cells take up Pi in a Na(d) manner. Uptake was slightly (to 0.78-fold of the control) decreased by 100 nmol/l dex but not affected by 0.1 mmol/l cAMP treatment. Although NaPi-IIb mRNA expression was maintained to some extent by AT II cells kept in primary culture, Pi uptake was more closely related to Pit-1 mRNA expression. PMID- 19806402 TI - Time interval and the factors associated with the development of asthma in patients with allergic rhinitis. AB - Rhinitis and asthma commonly coexist and studies have shown a positive association between rhinitis and asthma in both atopic and nonatopic adults. Longitudinal studies have shown that in many cases rhinitis precedes the onset of asthma. The aims of this study were to study the time interval for the development of asthma after the onset of rhinitis, to determine the proportion of patients in whom rhinitis precedes asthma, and to study the factors associated with the development of asthma in patients with allergic rhinitis compared to patients who continue to have allergic rhinitis alone. This was a cross-sectional study done at a tertiary care allergy center in Mysore, South India. It included consecutive patients between 2004 and 2006 with allergic rhinitis and/or asthma. We used a structured questionnaire, clinical evaluation, spirometry, and skin prick testing. A total of 1,141 subjects were included in the study. Among them, 700 had allergic rhinitis for varying intervals before developing asthma and 355 had rhinitis without asthma. In subjects aged 20 years or younger, logistic regression analysis confirmed an independent association with a family history of allergic rhinitis and sensitization to house dust mites as risk factors and ever used nasal steroids as protective against developing asthma in subjects with allergic rhinitis. In subjects older than 20 years, a family history of allergic rhinitis, atopy, and sensitization to house dust mites and trees were risk factors and ever-user of nasal steroids was protective. Rhinitis often preceded asthma and a high proportion of patients, both children and adults, developed asthma within 2 years after the onset of rhinitis. A family history of allergic rhinitis, atopy, and sensitization to house dust mites and trees are associated with the development of asthma in patients with allergic rhinitis. PMID- 19806404 TI - Two-step in vivo tumor targeting by biotin-conjugated antibodies and superparamagnetic nanoparticles assessed by magnetic resonance imaging at 1.5 T. AB - PURPOSE: The purpose of this study was to assess two-step in vivo tumor targeting by specific biotin-conjugated antibodies and ultrasmall superparamagnetic iron oxide (USPIO)-anti-biotin nanoparticles as contrast agents for magnetic resonance imaging (MRI) at 1.5 T. PROCEDURES: D430B human lymphoma cells, expressing the CD70 surface antigen, were injected either s.c. or i.v. to induce pseudo metastases in NOD/SCID mice. Thirty micrograms of biotin-conjugated monoclonal anti-CD70 was injected i.v., followed 4 h later by 8 micromol Fe/Kg USPIO-anti biotin. After 24 h, MRI was performed on T2* and b-FFE sequences. Signal intensity (SI) was calculated before and after USPIO-anti-biotin administration. RESULTS: Subcutaneous xenografts showed a dishomogeneous 30% decrease in SI on T2* with anti-CD70 + USPIO-anti-biotin treatment. Pseudo-metastatic xenografts showed a slight reduction in SI on T2*, but a 60% decrease in SI on b-FFE weighted sequences. Prussian blue staining confirmed the presence of iron nanoparticles in the excised tumors. CONCLUSION: MRI at 1.5 T can detect tumors by a two-step in vivo biotin-based protocol, which may allow the targeting of any cell surface antigen. PMID- 19806405 TI - Detection of experimentally induced pulmonary granuloma inflammation in monocyte chemoattractant protein-1 reporter mice. AB - PURPOSE: Among different chemokines, monocyte chemoattractant protein-1 (MCP-1) plays an important role in inflammatory disorders of lung. In response to stimuli, MCP-1 increases its transcription as an immediate early gene. In this paper, we describe the MCP-1-enhanced green fluorescent protein(EGFP) transgenic mouse in which EGFP expression is driven by human MCP-1 promoter and mimics the MCP-1 expression in situ. Thus, the MCP-1 reporter mouse model is designed to facilitate a better understanding of its role in various diseases. We employed this mouse model in a pulmonary granulomatous inflammation model using intratracheal instillation of Sephadex (SDX) beads and compared the EGFP reporter expression to endogenous MCP-1 expression through the course of inflammation. PROCEDURES: We analyzed the temporal pattern of SDX-induced infiltration of inflammatory cells in lung and in bronchoalveolar lavage fluid (BALF). The changes in tissue fluorescence, gene, and protein expressions for both MCP-1 and EGFP were analyzed. RESULTS: SDX instillation caused massive infiltration of inflammatory cells in BALF and lung tissue at the end of day 3. There was an increase of fluorescence in SDX-treated lung and BALF cells. By using lipopolysaccharide-induced systemic inflammation model, increase of fluorescence was found in bone marrow Gr-1(+) cells with high Mac-1 expression. MCP-1 and EGFP gene expression and MCP-1 protein level were increased after day 1, peaked at day 3, and declined toward basal levels at day 5. In contrast, EGFP protein level peaked after day 3 and remained elevated after day 5. Immunohistochemical staining revealed the MCP-1 and EGFP expression primarily at alveolar macrophages, macrophages infiltrating the granulomatous lesions and in bronchiolar epithelial cells. CONCLUSIONS: By using a pulmonary granuloma model, we showed that EGFP transgene reporter expression in MCP-1-EGFP mouse was correlated to the endogenous MCP-1 induction. The establishment of this mouse model will provide a valuable tool for monitoring the activation of monocytes/macrophages and facilitate the studies on the roles of MCP-1 gene in various inflammatory diseases. PMID- 19806406 TI - Quantitative assessment of the hepatic metabolic volume product in patients with diffuse hepatic steatosis and normal controls through use of FDG-PET and MR imaging: a novel concept. AB - AIMS AND OBJECTIVES: The aim of this study was to compare hepatic standardized uptake values (SUVs) and hepatic metabolic volumetric products (HMVP) between patients of diffuse hepatic steatosis and control subjects with normal livers. MATERIALS AND METHODS: Twenty-seven subjects were included in the study (13 men and 14 women; age range, 34-72 years). All had 18F-2-fluoro-2-D-deoxyglucose positron emission tomography (FDG-PET) and magnetic resonance imaging (MRI) scans with an interscan interval of 0-5 months. Twelve of 27 subjects had diffuse hepatic steatosis on MRI. The remaining 15 were selected as age-matched controls based on normal liver parenchyma on MRI. Mean and maximum hepatic SUVs were calculated for both patient groups on FDG-PET images. Hepatic volumes were measured from MRI. HMVP in each subject was subsequently calculated by multiplication of hepatic volume by mean hepatic SUV. HMVPs as well as mean and maximum hepatic SUVs were compared between the two study groups. RESULTS: HMVPs, mean hepatic SUVs, and maximum hepatic SUVs were greater (statistically significant, p < 0.05) in subjects with diffuse hepatic steatosis compared to those in the control group. CONCLUSION: The increase in HMVP is the result of increased hepatic metabolic activity likely related to the diffuse hepatic steatosis. The active inflammatory process related to the diffuse hepatic steatosis is the probable explanation for the increase in hepatic metabolic activity on FDG-PET study. PMID- 19806407 TI - The Clinical Application of Kaplan's Cardinal Line as a Surface Marker for the Superficial Palmar Arch. AB - We feel the original description of Kaplan's cardinal line provides a more accurate reference point to the superficial palmar arterial arch. We sought to anatomically correlate the relationship of Kaplan's cardinal line to the superficial palmar arch. Sixty hands (30 cadavers) were dissected after Kaplan's original description was drawn on each hand. Measurements we made from Kaplan's cardinal line to the superficial palmar arch at both the radial and ulnar borders of the ring finger. The superficial palmar arterial arch was an average of 10.4 and 11.8 mm from the radial and ulnar borders of the ring finger with standard deviations of roughly 4 mm for each measurement. Clinically, Kaplan's cardinal line is a more predictable landmark for the superficial palmar arch. In referencing this landmark as the distal most extent of an open or endoscopic carpal tunnel release, the superficial palmar arch should be free of transection. PMID- 19806408 TI - Free Muscle Transfer in Posttraumatic Plexopathies Part II: The Elbow. AB - The indications for free muscle transfer in brachial plexopathies are prolonged denervation time or inadequate upper extremity function after primary nerve reconstruction. The purpose of this study is to analyze the outcomes of free muscle transfer for elbow flexion and extension in brachial plexopathies in relation to the different muscles used and the respective motor donors. Seventy three muscles were transferred for elbow flexion and ten for elbow extension. Latissimus dorsi (LD) was used in 37 cases, gracilis in 28, rectus femoris (RF) in seven, and vastus lateralis in one. Five LD and five gracilis were transferred for elbow extension. Patients younger than 15 years yielded better results than older patients for elbow flexion. When LD was transferred, the mean muscle grading (MG) was 3.33 +/- 0.25 when the neurotization was from intercostals; these outcomes were statistically significant when compared with outcomes of free gracilis transfer (MG 2.25 +/- 0.6). There was also a statistically significant difference when free LD was neurotized with three intercostals as compared with two intercostals nerves. RF yielded also good results when neurotized from contralateral C7 (cC7; MG 3.67 +/- 0.6). For elbow extension, the better outcomes of LD were not statistically significant. Among all the free muscle transfers for upper extremity reconstruction, elbow reanimation yielded the most rewarding outcomes. The selection of powerful muscle units was more important than the effect of neurotization which was not as strong as it was in muscle transfers for facial or hand reanimation. PMID- 19806409 TI - Preoperative nomogram to predict risk of perioperative mortality following pancreatic resections for malignancy. AB - INTRODUCTION: The majority of pancreatic resections for malignancy are performed in older patients with major comorbidities. The aim of this study was to develop a preoperative nomogram based on the presence of comorbidities to predict risk of perioperative mortality. MATERIALS AND METHODS: The National Inpatient Sample database was queried to identify patients that underwent pancreatectomy for malignancy. The preoperative comorbidities identified as predictors were used, and a nomogram was created. Sample A (2000-2004) was utilized to develop the model, and sample B (2005) was utilized to validate this model. RESULTS: The overall actual observed perioperative mortality rate for samples A and B was 6.3% and 5.2%, respectively. The mean total points calculated for sample A by the nomogram was 131.7 that translates to a nomogram-predicted mortality rate of 4.9%, which is similar to the actual mortality. The mean total points for sample B was 128.1, which translates to a nomogram-predicted mortality rate of 4.6%. The similarity of mortality rates as predicted by the nomogram and a concordance index of 0.76 shows good agreement between the data and the nomogram. CONCLUSION: This preoperative nomogram has been shown to accurately predict the risk of perioperative mortality following pancreatectomy for malignancy. PMID- 19806410 TI - Occurrence and care of an obese patient during a serious adverse event with propofol for an endoscopy procedure. PMID- 19806411 TI - Tantalum is a good bone graft substitute in tibial tubercle advancement. AB - BACKGROUND: Porous tantalum is reportedly a good substitute for structural bone graft in several applications. So far, its use has not been reported in tibial tuberosity anteriorization (TTA) for treatment of isolated degenerative chondral lesions of the patellofemoral joint. QUESTIONS/PURPOSES: We asked whether the use of this material would produce similar standardized functional scores, pain (VAS), fusion rates, complications, and patient satisfaction to those for bone graft. PATIENTS AND METHODS: We performed a randomized, controlled trial in 101 patients (108 knees) scheduled for TTA comparing a porous tantalum implant (57 knees) with an autologous local tibial bone graft (51 knees). The minimum followup was 5 years (mean, 6.2 years; range, 5-8 years). RESULTS: At the last followup, clinical scores, fusion rates, and maintenance of the anteriorization either were better or similar for the TTA using the tantalum implant depending on the respective parameter. The operative technique was easier and shorter with the tantalum device. Complication and failure rates were greater using bone graft. Patient satisfaction was greater using the tantalum implant. CONCLUSIONS: Porous tantalum provided a reasonable alternative to bone graft in TTA. LEVEL OF EVIDENCE: Level I, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19806412 TI - Chest wall mass in a 50-year-old woman. PMID- 19806413 TI - The role of spirituality in the relationship between religiosity and depression in prostate cancer patients. AB - PURPOSE: This study aims to develop a theoretical framework of the relationship among religiosity, spirituality, and depression, potentially explaining the often mixed and inconsistent associations between religiosity and depression. METHODS: In this cross-sectional study, 367 men (average age of 66 +/- 9 years) with prostate cancer completed measures of religiosity (extrinsic/intrinsic), spirituality (Functional Assessment of Chronic Illness Therapy Spiritual Well Being Scale), quality of life (FACT-G), and depression (Hospital Anxiety and Depression Scale). RESULTS: There was a small relationship between intrinsic religiosity and depression (r = -0.23, p < 0.05) but a strong association between spirituality and depression (r = -0.58, p < 0.01). Using a mediation model, the meaning/peace subscale of the spirituality measure mediated the relationship between intrinsic religiosity and depression. This model controlled for age, marital status, stage of disease, time since diagnosis, hormone therapy, quality of life, and anxiety. CONCLUSIONS: When examining religiosity and spirituality, the main component that may help reduce depression is a sense of meaning and peace. These results highlight the potential importance of developing a patient's sense of meaning through activities/interventions (not exclusive to religious involvement) to achieve this goal. PMID- 19806414 TI - Can a motivational intervention overcome an unsupportive environment for walking- findings from the Step-by-Step Study. AB - BACKGROUND: Interventions to promote walking have rarely examined how their effects varied by the attributes of the physical environment. PURPOSE: The purpose of this study is to examine whether perceptions of environmental walkability predicted change in walking behavior following an individual-based intervention to promote walking and whether the intervention buffered the effects of unsupportive environment for walking. METHODS: Inactive adults (aged 30-65 years, 85% women) who completed a 3-month randomized control trial comparing the effect of a single mail-out of a theoretically based self-help walking program (WP, n = 102); the same program plus a pedometer (WPP, n = 105); and a "no treatment" control group (C, n = 107). Measures included change in self-reported walking time for all purposes and in the proportion of people reporting regular walking (i.e., > or =150 min/week and > or =5 sessions/wk). Perceptions of environmental esthetics, safety from crime, proximity to destinations, access to walking facilities, traffic, streetlights, connectivity, and hilliness were assessed at baseline and dichotomized into "low" or "high" by the median score. Covariates were social support, self-efficacy, intention to change behavior, and sociodemographic characteristics. RESULTS: Adjusting for baseline walking, significant covariates, and study groups, walking time at follow-up was lower if streetlights or esthetics were perceived to be "low" (-24% and -22%, respectively) compared with "high" (p < 0.05). In "low" esthetic conditions, those in the WPP were significantly more likely than controls to increase total walking time (Exp (b) = 2.53, p < 0.01) and to undertake regular walking (OR = 5.85, 95% CI 2.60-12.2), whereas in esthetically pleasing environments, the between-group differences were nonsignificant. CONCLUSIONS: Walkability attributes can influence individual-based walking programs. Some environmental barriers for walking can be overcome by motivational aids. PMID- 19806415 TI - Marital status and heart rate variability in patients with suspected coronary artery disease. AB - BACKGROUND: Married individuals are at reduced risk of cardiovascular disease morbidity and mortality. Recent research indicates that impaired heart rate variability (HRV) may contribute to cardiovascular disease morbidity and mortality and has also been associated with social isolation. PURPOSE: We investigated associations between HRV and marital status in patients with suspected coronary artery disease (CAD). METHODS: Eighty-eight patients who were being investigated for suspected CAD (28 women, 60 men, mean age 61.6, 60% married) were recruited from three rapid access chest pain clinics in London. Heart rate variability was measured using 24-h electrocardiograms and analyzed using frequency and time-domain measures. RESULTS: Unmarried marital status was associated with reduced heart rate variability as indexed by both frequency and time-domain measures, independently of age, gender, beta-blocker use, depression ratings, and subsequent diagnosis of significant CAD. CONCLUSION: These findings suggest that reduced heart rate variability is associated with not being married and may contribute to the reliably observed relationship between marital status and cardiovascular disease morbidity and mortality. PMID- 19806416 TI - Methodological issues in research on web-based behavioral interventions. AB - BACKGROUND: Web-based behavioral intervention research is rapidly growing. PURPOSE: We review methodological issues shared across Web-based intervention research to help inform future research in this area. METHODS: We examine measures and their interpretation using exemplar studies and our research. RESULTS: We report on research designs used to evaluate Web-based interventions and recommend newer, blended designs. We review and critique methodological issues associated with recruitment, engagement, and social validity. CONCLUSIONS: We suggest that there is value to viewing this burgeoning realm of research from the broader context of behavior change research. We conclude that many studies use blended research designs, that innovative mantling designs such as the Multiphase Optimization Strategy and Sequential Multiple Assignment Randomized Trial methods hold considerable promise and should be used more widely, and that Web-based controls should be used instead of usual care or no-treatment controls in public health research. We recommend topics for future research that address participant recruitment, engagement, and social validity. PMID- 19806417 TI - Prediction of BMI change in young children with the family nutrition and physical activity (FNPA) screening tool. AB - BACKGROUND: Youth obesity prevention practices would be enhanced if modifiable risk factors can be identified before children become overweight. PURPOSE: This study evaluated the predictive validity of the Family Nutrition and Physical Activity (FNPA) screening tool, a behaviorally based screening tool designed to assess family environments and behaviors that may predispose youth to becoming overweight. METHODS: Parents from a large urban school district completed the FNPA screening when children were in first grade. One-year change in measured body mass index (BMI) was used as the primary outcome, and this was computed using the relative change in distance from the BMI value at the 50th percentile. Descriptive, correlation, and mixed modeling analyses were used for survey validation. RESULTS: Over half of the participants exhibited an increase in BMI percentile over the 1-year follow-up with an average change of 0.51 +/- 11.5% which is indicative of trends to overweight. Although baseline BMI predicted BMI at follow-up, the FNPA total score explained unique variance in child BMI at follow-up after accounting for baseline BMI, parent BMI, and other demographic variables (p = 0.049). CONCLUSIONS: This study demonstrates the potential validity of a simple, easy-to-use screening tool for identifying children that may be at risk for becoming overweight. PMID- 19806418 TI - The vexing problem of post-amputation pain: what is the optimal perioperative pain management for below-knee amputation? PMID- 19806419 TI - [Ultrasound technique to locate the first tracheal ring]. PMID- 19806420 TI - Optimizing preoxygenation prior to tracheal intubation. PMID- 19806421 TI - Laparoscopic repair of Amyand's hernia in an 8-week-old infant. AB - Amyand's hernia is a rare type of hernia in which the appendix is found in the inguinal canal. A case of an 8-week-old boy who was diagnosed and managed laparoscopically is presented. To the authors' knowledge, this is the first report of emergency laparoscopic surgery for an Amyand's hernia in a young infant. PMID- 19806422 TI - Subsequent abdominal surgery after laparoscopic ventral and incisional hernia repair with an expanded polytetrafluoroethylene mesh: a single institution experience with 72 reoperations. AB - PURPOSE: Laparoscopic ventral and incisional hernia repair (LVIHR) carries a risk of adhesion formation and can influence subsequent abdominal operations (SAOs). We performed a retrospective study of findings during reoperations of patients who had previously had an LVIHR by using an expanded polytetrafluoroethylene mesh (DualMesh; WL Gore, Flagstaff, AZ, USA). METHODS: The medical records of all 695 patients who had LVIHR at our hospital were reviewed. Patients who underwent SAO for various indications were identified (n = 72) and analyzed. RESULTS: Seven LVIHR patients (1%) had early SAO (within a few days). In six patients (86%), removal of the mesh was required. Intra-operatively, in all six of these patients with peritonitis, there were no adhesions against the implant identified. Late SAOs (after more than 1 month) were performed in 65 patients (9.4%). Only one patient required acute surgical intervention due to an LVIHR-related adhesion (0.15%). Laparoscopy was performed in 83% and laparotomy in 17% of patients. Adhesions against the implant were present in 83% of patients; in 65%, the adhesions involved omentum only, and in 18%, they involved the bowel. Adhesiolysis was always easy and caused no inadvertent enterotomies. SAOs were devoid of postoperative complications. CONCLUSIONS: In this largest series of reoperations after LVIHR, the majority of patients had mild or moderate adhesions against the implant. The specific observations that: (1) no relaparoscopies had to be converted, (2) no inadvertent enterotomies were made during adhesiolysis, and (3) SAOs have practically been devoid of peri- and postoperative complications indicate that SAOs can be safely performed after previous LVIHR with DualMesh. PMID- 19806423 TI - A very simple technique to repair Grynfeltt-Lesshaft hernia. AB - PURPOSE: A very simple technique to repair a superior lumbar hernia is described. The location of this type of hernia, also known as the Grynfeltt-Lesshaft hernia, is defined by a triangle placed in the lumbar region. METHOD: An unusual case of a 67-year-old woman with a superior lumbar hernia is reported. The diagnosis was made by physical examination. RESULTS: The defect of the posterior abdominal wall was repaired with a polypropylene dart mesh. The patient had no evidence of recurrence at 11 months follow up. CONCLUSIONS: The surgical approach described in this paper is simple and easy to perform, and its result is comparable with other techniques that are much more sophisticated. No cases on the use of dart mesh to repair Grynfeltt-Lesshaft hernia have been reported by surgical journals indexed in PubMed. PMID- 19806424 TI - How do different diagnostic criteria, age and gender affect the prevalence of attention deficit hyperactivity disorder in adults? An epidemiological study in a Hungarian community sample. AB - The goal of the study was twofold: (1) to investigate the effect of different diagnostic criteria on prevalence estimates of adult attention deficit hyperactivity disorder (ADHD), and (2) to provide prevalence estimates of adult ADHD for the first time in a Hungarian sample. Subjects between 18 and 60 years were included in the screening phase of the study (N = 3,529), conducted in 17 GP practices in Budapest. Adult self-report scale 6-item version was used for screening. Out of 279 positively screened subjects 161 subjects participated in a clinical interview and filled out a self-report questionnaire to confirm the diagnosis. Beside DSM-IV diagnostic criteria, we applied four alternative diagnostic criteria: 'No-onset' (DSM-IV criteria without the specific requirement for onset); full/Sx (DSM-IV "symptoms only" criteria); and reduced/Sx (DSM-IV "symptoms only" criteria with a reduced threshold for symptom count). Crude prevalence estimates adjusted for the specificity and sensitivity data of the screener were 1.35% in the 'DSM-IV' group, 1.64% in the 'No-onset' group, 3.65% in the 'Sx/full' group and 4.16% in the 'Sx/reduced' group. Logistic regression analysis showed that ADHD was significantly more prevalent with younger age and male gender [chi(2) = 14.46; P = 0.0007]. Prevalence estimates corrected for the 'not-interviewed' subsample and adjusted for specificity and sensitivity data of the screener was 2.3% in males, 0.91% in females; 2.02% in the < or =40 years age group and 0.70% in the >40 years age group, based on DSM-IV diagnostic criteria. Prevalence rates found in this study are somewhat lower, but still are in line with those reported in the literature. PMID- 19806425 TI - Utility of the Japanese GFR estimation equation for evaluating potential donor kidney function. AB - BACKGROUND: In Japan, the number of living kidney transplantations has increased each year, and an accurate evaluation of renal function must be conducted before donation to minimize the risk to donors. Recently, the Japanese Society of Nephrology issued a new equation for estimating glomerular filtration rate (eGFR) in Japanese people. This study compared the accuracy of eGFR and creatinine clearance (Ccr) values with that of inulin clearance (Cin) for assessing renal function in kidney donors. METHODS: Clinical data were analyzed for 85 potential living kidney donors who had undergone routine measured GFR (mGFR) and Ccr measurements from October 2006 to November 2008 at a single center. Inulin clearance, representing the mGFR, was determined by standard method. The eGFR was calculated as: eGFR = 194 x Scr(-1.094) x Age(-0.287) (for females, x0.739). RESULTS: Mean mGFR was 96.1 +/- 14.7 (range 67.8-126.8); mean eGFR, 72.6 +/- 12.7 (range 50.1-107.1); and mean Ccr, 117.3 +/- 22.4 (range 35.1-170.1), in units of ml/min/1.73 m(2) for each. Relative to mGFR, the correlation coefficient for Ccr was 0.496, and the mean difference between the two values was 21.1 ml/min/1.73 m(2) (23.2%), with a root-mean square error (RMSE) of 19.6. The correlation coefficient between eGFR and mGFR was 0.502, and the mean difference between the two values was -23.5 (23.7%), with a RMSE of 11.0. Bland-Altman plots showed that Ccr overestimated mGFR in 90.6% of cases, whereas eGFR underestimated mGFR in 95.3% of cases. CONCLUSION: Ccr and eGFR values did not accurately estimate mGFR in Japanese living kidney donors. PMID- 19806426 TI - Effect of nonsteroidal anti-inflammatory drug plasters for knee osteoarthritis in Japanese: a randomized controlled trial. AB - The objective of this study was to examine the effect of a nonsteroidal anti inflammatory drug (NSAID) plaster for knee osteoarthritis among Japanese patients. An open-labeled, randomized, controlled, multiclinic trial was performed involving outpatient clinic groups. Two comparative groups-plaster NSAIDs and oral NSAIDs-were randomly allocated. The drugs used were limited to the current top three in both groups in Japan. Treatments were assessed after four weeks and compared with the baseline scores. Outcomes were evaluated by two psychometric measures: Japanese knee osteoarthritis measure, and pain with the visual analogue scale. The total number of patients included in the final evaluation was 165 (87 for the plaster group and 78 for the oral group). Between these two groups there were no significant differences in gender, age, body height and weight, body mass index, and X-ray grading. The subjects in both groups showed improvements in both scores at the end of intervention. The differences in the improvements in scores between the two groups were not significant, though the mean rank score and the 95% CI of the plaster group were slightly better than those of the oral group. In conclusion, the local application of a plaster with NSAIDs leads to the same level of improvement in knee osteoarthritis as oral NSAIDs. PMID- 19806427 TI - Prediction of hormone sensitivity for breast cancers. AB - The classic action that leads to transcriptional activation of estrogen response genes mediated through estrogen receptors (ER) and the estrogen complex plays a pivotal role in the development of ER-positive breast cancers. In addition to this pathway, non-classic action and non-genomic action, both estrogen-dependent and estrogen-independent genomic actions have also been found to contribute to ER positive tumor growth. Although the details of these mechanisms are not well known, participation of the growth factor signaling pathway is likely to be the most significant factor for acquisition of resistance to hormonal therapy. This resistance is mediated not only directly through cell growth promotion by growth factor signaling, but also through enhancement of alternative ER signaling pathways in addition to classic action. The reason why tamoxifen-insensitive ER positive breast cancers respond to aromatase inhibitors may be explained, at least in part, by the different estrogen-related signaling pathways in which aromatase inhibitors may block estrogen signaling. In this paper we discuss the molecular mechanisms for resistance to hormonal therapy based on an understanding of estrogen signaling pathways. PMID- 19806428 TI - Breast cancer stem cells. AB - Since the initial discovery of leukemia stem cells nearly a decade ago, a great deal of cancer research has focused on the identification of cancer stem cells (CSCs) in many types of solid tumors, including breast cancer. Through analysis of cell surface markers and xenotransplant models, a subpopulation of putative human breast cancer stem cells (BCSCs) that is CD24-negative/CD44-positive (CD24( )/CD44+) and bears high aldehyde dehydrogenase 1 activity has been isolated in clinical samples of breast cancer tissues. Human BCSCs are considered to be derived from basal cells that reside in the basal membranes of alveolar units in human adult mammary glands. Furthermore, BCSCs have been shown to express higher levels of oxidative stress-responsive genes, which could confer part of their ability to resist anti-cancer therapy, than non-CSCs. The emerging picture of the biological properties of BCSCs would contribute for devising innovative therapies for breast cancer, targeting the intrinsic and extrinsic factors that maintain the BCSCs. PMID- 19806429 TI - Three cases of kindred with familial breast cancer in which carrier detection by BRCA gene testing was performed on family members. AB - We performed presymptomatic carrier detection by BRCA gene testing of the family members of three familial breast cancer kindred diagnosed with pathogenetic mutation in BRCA genes. All members were over 20 years of age. We explained familial breast cancer and BRCA gene testing, and obtained autonomic consent before gene testing. Genetic testing revealed twins in a family were dizygotic. In another family bilateral breast cancer occurred in a carrier after five years of genetic testing. Carriers are at high risk of breast cancer and have to receive breast cancer screening and familial tumor counseling. Non-carriers are at the same risk of breast cancer as the general population. PMID- 19806430 TI - Development of a new disinfectant with very strong anti-influenza viral activity: a preliminary report. AB - OBJECTIVES: We evaluated the effectiveness and safety of a disinfectant newly developed by our laboratories for use against influenza viruses. METHODS: The effectiveness of our new disinfectant against avian, swine and human influenza viruses was tested in ovo. The acute toxicity of this disinfectant to two different cultured cell lines was investigated. RESULTS: This new disinfectant showed very strong anti-influenza viral activity in the in ovo tests. All of the influenza viruses tested were inactivated very quickly. Following exposure to the disinfectant, the infectivity of all viral strains tested had been eliminated within <=10 min. The infectant showed a weak acute toxicity in vitro. CONCLUSION: This new disinfectant is expected to be useful for preventing viral infection during a new influenza pandemic. PMID- 19806431 TI - Description of environmental determinants of quality of life in children with intellectual disability in Japan using the Delphi technique. AB - OBJECTIVES: The present study aimed to define the framework of an environment conducive to the well-being of children with intellectually disability (CID). METHODS: A questionnaire composed of 31 items was developed through literature review. Then a 2-round Delphi survey was conducted with 3 different panels: health professionals (HPs), parents of CID, and teachers. The participants were asked to rate each item, select and rank the 10 most important items, and suggest additional ones. RESULTS: A total of 71 participants responded to the first round: 24 HPs, 22 parents, and 25 teachers. In the second round the overall response rate was 83%. At the end of the exercise, 12 items reached global consensus, i.e., in all groups. Only 5 items were ranked as most important by all groups: attitudes of family members at home; attitudes of HPs and teachers; support from family members at home; support at school (classmates and teachers); and government policies. Nevertheless, the panelists' views diverged on the remaining items. Several additional elements were suggested. CONCLUSIONS: The views of HPs, teachers, and parents are complementary for the improvement of quality of life (QOL) of CID. The present findings will be used as a basis for the development of an instrument to assess the living environment of CID. PMID- 19806433 TI - The influence of flow rate on the aerosol deposition profile and electrostatic charge of single and combination metered dose inhalers. AB - PURPOSE: The capability of the electrostatic next generation impactor (eNGI) has been investigated as a tool capable of measuring the electrostatic charge of single (Flixotide; containing fluticasone propionate (FP)) and combination (Seretide; FP and salmeterol xinafoate (SX)) pressurised metered dose inhalers (pMDIs) at different flow rates. METHODS: Aerosol mass distributions were investigated at 30, 60 and 90 l.min(-1) and simultaneous charge measurements recorded. RESULTS: Analysis of the mass distribution data indicated a flow dependent relationship, where the aerosol performance (aerodynamic diameter <5 mum) of FP significantly increased between 30 l.min(-1) and 60 l.min(-1) for both formulations. No significant increase in SX was observed for Seretide with increased flow rate. Analysis of the charge distribution indicated both formulations to primarily charge negatively with a concurrent increase in charge with increased flow rate. Interestingly, the charge-tomass ratio remained relatively constant between 30 l.min(-1) and 60 l.min(-1) and increased at 90 l.min(-1), indicating that charging was majorly influenced at the highest flow rate. CONCLUSIONS: This study has shown how the eNGI could be used as a simple Pharmacopeia based methodology for the evaluation of mass and charge profiles of single and combination pMDIs at a series of flow rates. PMID- 19806432 TI - Fructose and moderately high dietary salt-induced hypertension: prevention by a combination of N-acetylcysteine and L-arginine. AB - Diets containing 8% salt or 4% fructose (FR) cause insulin resistance and increase tissue methylglyoxal and advanced glycation end products (AGEs), platelet cytosolic-free calcium, and systolic blood pressure (SBP) in rats. In WKY rats, we have shown that moderately high salt, 4% NaCl (MHS) alone in diet does not cause hypertension, and when given along with 4% FR it does not have an additive effect. N-acetylcysteine (NAC) or L-arginine (ARG), treatment alone does not prevent hypertension in this model. The objectives of this study were to investigate the effect of NAC plus ARG in diet on SBP, platelet cytosolic-free calcium in a MHS + FR model, and to measure the plasma levels of methylglyoxal and the AGE, methylglyoxal-derived hydroimidazolone (MGH). At 7 weeks of age, WKY rats were divided into three groups: control group was given regular rat chow (0.7% NaCl) and water; MHS + FR group, diet containing 4% NaCl and 4% FR in drinking water; and MHS + FR + NAC + ARG group, MHS diet supplemented with 1.5% N acetylcysteine (NAC) and 1.5% L-arginine (ARG), and 4% FR in drinking water, and followed for 6 weeks. NAC + ARG prevented the increase in platelet cytosolic-free calcium and SBP in MHS + FR treated rats. There was no difference in mean values of plasma methylglyoxal and MGH among the groups. In conclusion, NAC + ARG treatment is effective in preventing hypertension in a moderately high salt + FR induced animal model. Plasma methylglyoxal and MGH may not represent tissue modification or, alternatively, other tissue AGEs, derived from methylglyoxal or other aldehydes, may be involved in hypertension in this model. PMID- 19806434 TI - Reverse iontophoresis of amino acids: identification and separation of stratum corneum and subdermal sources in vitro. AB - PURPOSE: To differentiate the stratum corneum (SC) and subdermal sources of amino acids (AAs) extracted by reverse iontophoresis. METHODS: 13 zwitterionic AAs were quantified in this in vitro study. Repetitive tape-stripping permitted the distribution of the analytes to be determined in the SC. Iontophoresis experiments were performed in which the subdermal chamber contained either phosphate-buffered saline (PBS) only, or a mixture of the 13 AAs in PBS. RESULTS: AAs were homogeneously distributed across the SC and broadly divided into three groups (high, medium, low) in terms of total amount present. As expected, extraction to the cathode for the essentially neutral analytes involved was more efficient. Initial samples obtained during the first hour of iontophoresis primarily extracted AAs from the SC. The fluxes observed in the latter half of the 6-h experiment, on the other hand, correlated well with the corresponding subdermal concentrations. CONCLUSION: A relatively short extraction period (approximately 1 h) by reverse iontophoresis can be used to evaluate the content of AAs in the SC. Once this 'reservoir' has been depleted, reverse iontophoresis can then monitor the subdermal concentrations of the AAs. The latter appears most useful for compounds which are present at lower levels in the SC. PMID- 19806435 TI - Effects of polymer type and storage relative humidity on the kinetics of felodipine crystallization from amorphous solid dispersions. AB - PURPOSE: The objective of this study was to investigate the effects of polymer type and storage relative humidity (RH) on the crystallization kinetics of felodipine from amorphous solid dispersions. METHODS: Crystallization of the model drug felodipine from amorphous solid dispersion samples containing poly(vinyl pyrrolidone) (PVP) and hypromellose acetate succinate (HPMCAS) were evaluated. Samples at three different drug-polymer weight ratios (10, 25, and 50 wt. % polymer) were prepared and stored at six different RHs (0%, 32%, 52% or 66%, 75%, 86%, and 93%). Periodically, the fraction of the drug that had crystallized from the samples was quantified using powder X-ray diffractometry (PXRD). RESULTS: Felodipine crystallization rates from PVP-containing dispersions were found to be very sensitive to changes in storage RH, while crystallization rates from HPMCAS-containing dispersions were not. PVP and HPMCAS were similar in terms of their ability to inhibit crystallization at low RH, but when the storage RH was increased to 75% or above, felodipine crystallization from PVP-containing solid dispersions proceeded much faster. It is hypothesized that this trend was caused by moisture-induced drug-polymer immiscibility in PVP-felodipine system. For PVP-containing solid dispersion samples stored at 75% RH and above, crystallization of the model drug felodipine seemed to approach a kinetic plateau, whereby a fraction of the drug still remained amorphous even after storage for 500 days or more. CONCLUSIONS: The physical stability of solid dispersions as a function of RH is highly dependent on the polymer used to form the solid dispersion, with PVP-containing dispersions being much less physically stable at high RH than HPMCAS-containing dispersions. PMID- 19806436 TI - Leucine-Aspartic Acid-Valine sequence as targeting ligand and drug carrier for doxorubicin delivery to melanoma cells: in vitro cellular uptake and cytotoxicity studies. AB - PURPOSE: To study the feasibility of Leucine-Aspartic Acid-Valine (LDV) as targeting ligand and drug carrier for targeted delivery to integrin alpha(4)beta(1) over-expressing cancer cells. METHODS: Poly(L,D,V) was randomly copolymerized using N-carboxyanhydrides of leucine, beta-benzyl-aspartic acid, and valine. Oligo(LDV), consisting of 2-6 LDV units, were synthesized by solid phase protein synthesis (SPPS) method. Binding of Leu-Asp-Val, Val-Asp-Leu, and Leu-Asn-Val, and internalization of FITC labeled LDV by wild-type and integrin alpha(4) knock-down A375 cells were studied. Cytotoxicity of poly(L,D,V)-Dox, oligo(LDV)-Dox, and doxorubicin (Dox) was also determined on wild-type, integrin alpha(4) knock-down A375 cells, and normal human epithelial keratinocytes (NHEK). RESULTS: LDV was essential for the specific binding and internalization by cells expressing integrin alpha(4)beta(1). Cytotoxicity of poly(L,D,V)-Dox and oligo(LDV)-Dox was integrin alpha(4)-dependent, while free Dox did not show this differential effect. No observable cytotoxicity trend was found when increasing LDV repeating unit. Poly(L,D,V) was relatively more effective than oligo(LDV) for the delivery of Dox to A375. CONCLUSION: LDV containing moieties bind specifically to integrin alpha(4)beta(1) expressing cancer cells. The binding, internalization, and cytotoxicity depend on the level of integrin alpha(4)beta(1) expression. Poly(L,D,V) and oligo(LDV) were both effective in the in vitro targeted delivery of Dox to integrin alpha(4)beta(1) over-expressing A375 cells. PMID- 19806437 TI - Adjusting rehabilitation costs and benefits for health capital: the case of low back occupational injuries. AB - INTRODUCTION: Case-mix adjustments for treatment/rehabilitation costs and benefits of non-traumatic injuries, such as occupational back pain, are much more difficult than adjustments for traumatic injuries. We present a new method for adjusting for severity differences in the costs and benefits of treating occupational low back injuries. METHODS: Using initial post-injury differences in the health capital of prospective sample of 1,831 occupational related back pain patients, we combine survey data with workers' compensation claim files and medical billing information to adjust the costs and benefits of treatment using multivariate techniques. RESULTS: We find that large differences in the net benefits of treatment between the three lowest cost provider groups virtually disappear once adjustments are made for worker's health capital (injury severity) at entry into treatment. CONCLUSIONS: Once adjustments are made for initial health capital immediately after injury, the net benefits of treating occupational low back pain are virtually identical for physician only care, physician plus physical therapy care, and chiropractic care. Net benefits of care are lower for combined physician/chiropractic care, and lowest for all other forms of care (principally, treatment by orthopedic surgeons). Our method is readily adapted for comparisons among individual health care/occupational rehabilitation professionals or among group practices and other health care organizations. PMID- 19806438 TI - Understanding decisions about work after spinal cord injury. AB - INTRODUCTION: Research has consistently shown that many people with spinal cord injury (SCI) do not return to work (RTW), despite evidence that being employed is associated with better quality of life, participation and physical and psychological well-being. While some factors associated with RTW outcome have been identified, very little is known about what influences people's own decisions about their employment following SCI. This qualitative study sought to identify factors that influenced decisions about whether and when to RTW for people with SCI. METHODS: Participants were recruited through rehabilitation and support services in Aotearoa/New Zealand, and sampling sought to capture variation in the population, particularly with regard to pre-injury occupation, physical impairment, age, ethnicity and education. In-depth interviews were conducted which were audio-taped and transcribed. Interviews were analysed using rigorous methods drawn from grounded theory. RESULTS: Thirteen participants took part in the research. Findings identified four main themes that synthesised experiences about what influenced decisions about employment after SCI: (1) ability to work (given work demands and current resources); (2) presence of responsibilities or pressures that compete with work; (3) access to a suitable job; and (4) whether work was of enough benefit to the individual to be worth pursuing. CONCLUSIONS: Findings illustrate the complexity of decisions about working after SCI, and provide a framework that health and vocational professionals may find useful to inform discussions with their clients. Findings also provide a basis for further research into interventions to support people to make informed decisions about employment after SCI. PMID- 19806439 TI - Multi label learning for prediction of human protein subcellular localizations. AB - Predicting protein subcellular locations has attracted much attention in the past decade. However, one of the most challenging problems is that many proteins were found simultaneously existing in, or moving between, two or more different cell components in a eukaryotic cell. Seldom previous predictors were able to deal with such multiplex proteins although they have extremely important implications in future drug discovery in terms of their specific subcellular targeting. Approximately 20% of the human proteome consists of such multiplex proteins with multiple sample labels. In order to efficiently handle such multiplex human proteins, we have developed a novel multi-label (ML) learning and prediction framework called ML-PLoc, which decomposes the multi-label prediction problem into multiple independent binary classification problems. ML-PLoc is constructed based on support vector machine (SVM) and sequential evolution information. Experimental results show that ML-PLoc can achieve an overall accuracy 64.6% and recall ratio 67.2% on a benchmark dataset consisting of 14 human subcellular locations, and is very powerful for dealing with multiplex proteins. The current approach represents a new strategy to deal with the multi-label biological problems. ML-PLoc software is freely available for academic use at: http://www.csbio.sjtu.edu.cn/bioinf/ML-PLoc . PMID- 19806440 TI - Modeling the 3-D structure of a recombinant laccase from Trametes trogii active at a pH close to neutrality. AB - A cDNA encoding a novel laccase from the white-rot fungus Trametes trogii was cloned and expressed in Pichia pastoris. The recombinant protein (Lcc2) exhibited kinetic parameters for both phenolic and non phenolic substrates that were different from the previously described Lcc1, the main laccase isoform expressed by T. trogii; in addition, the pH/activity profiles for phenolic substrates of Lcc2 were shifted upward by 1-1.5 pH units towards neutrality as compared to Lcc1. Comparative modeling of the two laccases (69.2% identity) showed that the overall fold of Lcc2 is very similar to Lcc1 and other laccases. The substrate cavity of Lcc2 contains the Asp residue which is thought to mediate the laccase activity at acidic pHs, whereas two hydrophobic residues (Phe, Ile) on the cavity orifice of Lcc2 replace the two polar residues (Thr, Ser) of Lcc1. These structural differences may be responsible for the unique kinetic performances of Lcc2. PMID- 19806443 TI - The moderating effects of parenting styles on African-American and Caucasian children's suicidal behaviors. AB - Given that parenting practices have been linked to suicidal behavior in adolescence, examining the moderating effect of parenting styles on suicidal behavior early in development could offer potential insight into possible buffers as well as directions for suicide prevention and intervention later in adolescence. Hence, the moderating effects of parenting styles, including authoritarian, permissive, and features of authoritative parenting, on depressed and aggressive children's suicidal behavior, including ideation and attempts, were evaluated with young children (N = 172; 72% male, 28% female) ranging from 6 to 12 years of age. African American (69%) and Caucasian (31%) children admitted for acute psychiatric inpatient care completed standardized measures of suicidal behavior, depressive symptoms, and proactive and reaction aggression. Their parents also completed standardized measures of parental distress and parenting style. Hierarchical regression analyses revealed that, while statistically controlling for age and gender, children who endorsed more depressive symptoms or reactive aggression reported more current and past suicidal behavior than children who endorsed fewer depressive or aggressive symptoms. The significant positive relationship observed between depressive symptoms and childhood suicidal behavior, however, was attenuated by parental use of authoritarian parenting practices for African-American and older children but not for younger and Caucasian children. The ethnic/racial difference observed for the buffering effect of authoritarian parenting practices offers potential theoretical and clinical implications for conceptualizing the moderating effects of parenting styles on African-American and Caucasian children's suicidal behavior. PMID- 19806442 TI - Amino acids variations in amyloid-beta peptides, mitochondrial dysfunction, and new therapies for Alzheimer's disease. AB - Soluble oligomers and/or aggregates of Amyloid-beta (Abeta) are viewed by many as the principal cause for neurodegeneration in Alzheimer's disease (AD). However, the mechanism by which Abeta and its aggregates cause neurodegeneration is not clear. The toxicity of Abeta has been attributed to its hydrophobicity. However, many specific mitochondrial cytopathologies e.g., loss of complex IV, loss of iron homeostasis, or oxidative damage cannot be explained by Abeta's hydrophobicity. In order to understand the role of Abeta in these cytopathologies we hypothesized that Abeta impairs specific metabolic pathways. We focused on heme metabolism because it links iron, mitochondria, and Abeta. We generated experimental evidence showing that Abeta alters heme metabolism in neuronal cells. Furthermore, we demonstrated that Abeta binds to and depletes intracellular regulatory heme (forming an Abeta-heme complex), which provides a strong molecular connection between Abeta and heme metabolism. We showed that heme depletion leads to key cytopathologies identical to those seen in AD including loss of iron homeostasis and loss of mitochondrial complex IV. Abeta heme exhibits a peroxidase-like catalytic activity, which catalytically accelerates oxidative damage. Interestingly, the amino acids sequence of rodent Abeta (roAbeta) and human Abeta (huAbeta) is identical except for three amino acids within the hydrophilic region, which is also the heme-binding motif that we identified. We found that huAbeta, unlike roAbeta, binds heme tightly and forms a peroxidase. Although, roAbeta and huAbeta equally form fibrils and aggregates, rodents do not develop AD-like neuropathology. These findings led us to propose a new mechanism for mitochondrial dysfunction and huAbeta's neurotoxicity. This mechanism prompted the development of methylene blue (MB), which increased heme synthesis, complex IV, and mitochondrial function. Thus, MB may delay the onset and progression of AD and serve as a lead to develop novel drugs to treat AD. PMID- 19806444 TI - Dimensionally-reduced visual cortical network model predicts network response and connects system- and cellular-level descriptions. AB - Systems-level neurophysiological data reveal coherent activity that is distributed across large regions of cortex. This activity is often thought of as an emergent property of recurrently connected networks. The fact that this activity is coherent means that populations of neurons may be thought of as the carriers of information, not individual neurons. Therefore, systems-level descriptions of functional activity in the network often find their simplest form as combinations of the underlying neuronal variables. In this paper, we provide a general framework for constructing low-dimensional dynamical systems that capture the essential systems-level information contained in large-scale networks of neurons. We demonstrate that these dimensionally-reduced models are capable of predicting the response to previously un-encountered input and that the coupling between systems-level variables can be used to reconstruct cellular-level functional connectivities. Furthermore, we show that these models may be constructed even in the absence of complete information about the underlying network. PMID- 19806445 TI - Psychotropic medication use among children with autism spectrum disorders enrolled in a national registry, 2007-2008. AB - Patterns of current psychotropic medication use among 5,181 children with autism spectrum disorders (ASD) enrolled in a Web-based registry were examined. Overall, 35% used at least one psychotropic medication, most commonly stimulants, neuroleptics, and/or antidepressants. Those who were uninsured or exclusively privately insured were less likely to use >or=3 medications than were those insured by Medicaid. Psychiatrists and neurologists prescribed the majority of psychotropic medications. In multivariate analysis, older age, presence of intellectual disability or psychiatric comorbidity, and residing in a poorer county or in the South or Midwest regions of the United States increased the odds of psychotropic medication use. Factors external to clinical presentation likely affect odds of psychotropic medication use among children with ASD. PMID- 19806446 TI - Few Italian drug users undergo HIV testing. AB - We estimated the proportion of drug users at treatment centres in Italy who had undergone HIV testing and the correlates of testing. Of the 1,917 drug injectors, 37.4% had been tested in the current year; of the 665 non-injectors, 28% had been tested. Among injectors, testing was associated with: being older than 35, foreign nationality, residing in central Italy, drug use for over 2 years, and having undergone both pharmacological and psychological treatment. Among non injectors, an association was found for foreign nationality and not having been treated at other facilities. The results stress the need to facilitate access to testing. PMID- 19806447 TI - LMTK2 and PARP-2 gene polymorphism and azoospermia secondary to meiotic arrest. AB - PURPOSE: To investigate whether the human LMTK2 and PARP-2 gene defects are associated with azoospermia by meiotic arrest, mutational analysis was performed on Japanese men with azoospermia. METHODS: Via direct sequencing, mutational screening was carried out on the exon region of the genes, using genomic DNAs from 18 Japanese men. Statistical analysis was done on the detected single nucleotide polymorphisms (SNPs) in the patients and normal controls. RESULTS: Nine SNPs were detected in LMTK2 and five SNPs were detected in PARP-2. There were no significant differences in the genotype distribution and allele frequencies between the two groups in LMTK2. However, the genotype frequency of heterozygotes in SNP1 of PARP-2 was higher in the patient group. The haplotype analysis revealed that SNP1-SNP4 (T-A) of PARP-2 was significantly more frequent in the patient group. CONCLUSION: The PARP-2 gene might be associated with azoospermia by meiotic arrest in humans. PMID- 19806448 TI - Intranasal delivery of human beta-amyloid peptide in rats: effective brain targeting. AB - (1) Intranasal administration is a non-invasive and effective way for the delivery of drugs to brain that circumvents the blood-brain barrier. The aims of the study were to test a nasal delivery system for human beta-amyloid (A beta) peptides, to measure the delivery of the peptides to brain regions, and to test their biological activity in rats. (2) A beta(1-42), in the form of a mixture of oligomers, protofibrils, and fibrils was dissolved in a nasal formulation containing hydrophobic, hydrophylic, and mucoadhesive components. The peptide solution was administered intranasally to rats as a single dose or in repeated doses. (3) Nasally injected A beta labeled with the blue fluorescent dye amino methyl coumarinyl acetic acid (AMCA) could be detected by fluorescent microscopy in the olfactory bulb and frontal cortex. The concentration of the peptide was quantified by fluorescent spectroscopy, and a significant amount of AMCA-A beta peptide could be detected in the olfactory bulb. Unlabeled A beta also reached the olfactory bulb and frontal cortex of rats as evidenced by intense immunostaining. (4) In behavioral experiments, nasal A beta treatment did not affect anxiety levels (open-field test) and short-term memory (Y-maze test), but significantly impaired long-term spatial memory in the Morris water maze. The treatments did not result in A beta immunization. (5) The tested intranasal delivery system could successfully target a bioactive peptide into the central nervous system and provides a basis for developing a non-invasive and cost effective, new model to study amyloid-induced dysfunctions in the brain. PMID- 19806449 TI - Reliable data on 5- and 10-year survival provide accurate estimates of 15-year survival in estrogen receptor-positive early-stage breast cancer. AB - There are few studies of model-based survival projections using early empirical results for estimating long-term survival. Utilizing Early Breast Cancer Trialists' Collaborative Group (EBCTCG) data, a Markov model was generated to compare empirical results with those modeled beyond the empirical result time horizon in estrogen receptor (ER)-positive early-stage breast cancer (ESBC). Modeling 15-year survival based on 5- and 10-year EBCTCG data resulted in an average error estimate in breast cancer mortality of 0.75% [range -0.83 to 2.19%]. Although modeling life expectancy differences ranged from an underestimate of -7.93% to an overestimate of 12.64%, over the span of 15 years this corresponded to a loss of 18 days or a gain of 40 days of life. Reliable early survival data may be used to generate models that accurately estimate 15 year survival in ER-positive ESBC. Whether early survival data can be employed over the lifetime horizon remains to be demonstrated. PMID- 19806450 TI - Overexpression of p53 is correlated with poor outcome in premenopausal women with breast cancer treated with tamoxifen after chemotherapy. AB - The aim of this study was to evaluate the difference in outcomes based on p53 overexpression of patients with breast cancer who received adjuvant therapy following local treatment for invasive ductal carcinoma, not otherwise specified. We analyzed data from 4,683 patients with cancer enrolled in two institutions between 1997 and 2006. We analyzed the correlation between p53 overexpression and relapse, response to adjuvant therapy, breast cancer-specific survival (BCSS), and relapse-free survival (RFS) in patients with primary breast cancer. Overexpression of p53 was noted in 1,091 patients (23.3%). A significant correlation existed between p53 overexpression and poor prognostic factors, an increased frequency of regional recurrence, visceral metastasis, and worse BCSS and RFS. Based upon subgroup analyses, combined age (<35, 35-50, and >50 years) and adjuvant therapy (hormone therapy only, chemotherapy only, and hormone therapy following chemotherapy), the greatest reduction of survival based on p53 overexpression was noted in patients 35-50 years of age who received hormone therapy following chemotherapy (P < 0.05). Multivariate analysis showed that p53 overexpression is an independent prognostic factor in patients treated with hormone therapy and chemotherapy (relative risk for BCSS, 2.003; 95% CI, 1.105 3.631; P = 0.022). The p53-overexpressing patients with breast cancer between 35 and 50 years of age who received tamoxifen following chemotherapy had the greatest adverse effect on outcome. Overexpression of p53 is significantly associated with tamoxifen resistance in premenopausal women with breast cancer. PMID- 19806451 TI - Analysis of the structure and neuritogenic activity of chondroitin sulfate/dermatan sulfate hybrid chains from porcine fetal membranes. AB - The amniotic membrane (AM) is the innermost layer of fetal membranes and possesses various biological activities. Although the mechanism underlying these biological activities remains unclear, unique components seem to be involved. AM contains various extracellular matrix components such as type I collagen, laminin, fibronectin, hyaluronan, and proteoglycans bearing chondroitin sulfate/dermatan sulfate (CS/DS) glycosaminoglycan side chains. Since CS/DS have been implicated in various biological processes, we hypothesized that CS/DS in AM may play a major role in the biological activities of AM. Therefore, the structure and bioactivity of the CS/DS chains from porcine fetal membranes (FM CS/DS) were investigated. A compositional analysis using various chondroitinases revealed that the characteristic DS domain comprised of iduronic acid-containing disaccharide units is embedded in FM-CS/DS, along with predominant disaccharide units, GlcA-GalNAc, GlcA-GalNAc(4-O-sulfate), and GlcA-GalNAc(6-O-sulfate), where GlcA and GalNAc represent D-glucuronic acid and N-acetyl-D-galactosamine, respectively. The average molecular mass of FM-CS/DS chains was unusually large and estimated to be 250 - 300 kDa. The FM-CS/DS chains showed neurite outgrowth promoting activity, which was eliminated by digestion with chondroitinase ABC of the CS/DS chains. This activity was suppressed by antibodies against growth factors including pleiotrophin, midkine, and fibroblast growth factor-2, suggesting the involvement of these growth factors in the neurite outgrowth promoting activity. The binding of these growth factors to FM-CS/DS was also demonstrated by surface plasmon resonance spectroscopy. PMID- 19806453 TI - Do genetically modified plants impact arbuscular mycorrhizal fungi? AB - The development and use of genetically modified plants (GMPs), as well as their ecological risks have been a topic of considerable public debate since they were first released in 1996. To date, no consistent conclusions have been drawn dealing with ecological risks on soil microorganisms of GMPs for the present incompatible empirical data. Arbuscular mycorrhizal fungi (AMF), important in regulating aboveground and underground processes in ecosystems, are the most crucial soil microbial community worthy of being monitored in ecological risks assessment of GMPs for their sensitivity to environmental alterations (plant, soil, climatic factor etc.). Based on current data, we suggest that there is a temporal-spatial relevance between expression and rhizosphere secretion of anti disease and insecticidal proteins (e.g., Bt-Bacillus thuringiensis toxins) in and outer roots, and AMF intraradical and extraradical growth and development. Therefore, taking Bt transgenic plants (BTPs) for example, Bt insecticidal proteins constitutive expression and rhizosphere release during cultivation of BTPs may damage some critical steps of the AMF symbiotic development. More important, these processes of BTPs coincide with the entire life cycle of AMF annually, which may impact the diversity of AMF after long-term cultivation period. It is proposed that interactions between GMPs and AMF should be preferentially studied as an indicator for ecological impacts of GMPs on soil microbial communities. In this review, advances in impacts of GMPs on AMF and the effect mechanisms were summarized, highlighting the possible ecological implications of interactions between GMPs and AMF in soil ecosystems. PMID- 19806452 TI - The effects of contaminants in European eel: a review. AB - European eel (Anguilla anguilla (L.)) stocks are in decline in most of their geographical distribution and their status is considered below safe biological limits. Recently, there is an increasing awareness that spawner quality might be an essential element in the decline of the species since pollution by bioaccumulating chemical substances may have a large impact on the reproduction success of the eel. This review gives an overview of the literature on the effects of contaminants on the European eel and on the consequences on the biology and fitness of the eel in order to document the role of pollution in its decline. A variety of contaminants have been found to affect the eel. These contaminants may cause disturbance of the immune system, the reproduction system, the nervous system and the endocrine system and effects were reported on several levels of biological organization, from subcellular, organ, individual up to even population level. More extensive research is needed in order to evaluate how pollutants are detrimental to eel populations. Getting a comprehensive overview of the quality (including contamination levels, biomarker responses, lipid content and condition) of the silver eel population all over Europe seems to be an essential and urgent objective for the European eel management. PMID- 19806454 TI - Avian liver organochlorine and PCB from South coast of the Caspian Sea, Iran. AB - Liver samples (n = 43) of 9 avian species representing the families Phalacrocoracidae, Podicipedidae, Laridae, and Anatidae, were collected from the Iranian coast of the Caspian Sea. Samples were analyzed for organochlorine pesticides (OCPs), such as dichlorodiphenyltrichloroethane (DDT) and its metabolites, hexachlorobenzene (HCB), hexachlorocyclohexane isomers (HCHs), and seven PCB congeners. p,p'-DDE was predominantly found in all species, at concentrations ranging from the limit of quantification (LOQ) to 340 ng/g ww. Most frequently encountered PCB congeners, in all samples, were 118, 153 and 138; and birds in Phalacrocoracidae had the highest liver PCB (mean 90 +/- 32; ranging from or =70 years of age. These studies demonstrate improved survival among optimally debulked patients. There are limited data specifically addressing the advanced elderly, age > or =80; however, selected patients appear to be appropriate candidates. Further dedicated study addressing the patient selection for debulking surgery and treatment outcomes among elderly ovarian cancer patients is needed. In this article, outcome studies addressing the application of major gynecologic surgery, including cytoreductive surgery, in the elderly population are reviewed, and current areas of clinical investigation are highlighted. PMID- 19806461 TI - Bioequivalence; its history, practice, and future. PMID- 19806462 TI - The influence of iron stores on cadmium body burden in a Thai population. AB - Cadmium is a toxin of increasing public health concern due to its presence in most human foodstuffs and in cigarette smoke. Exposure to cadmium leads to tissue bioaccumulation and, in particular, has nephrotoxic effects. The aim of the present study was to investigate the association between cadmium body burden and iron stores in a Thai population. A total of 182 healthy adult Thai subjects of both genders (89 males, 93 females) aged between 18 and 57 years and weighing 40 95 kg were included in this study. The total amounts of cadmium excreted in urine over 2 h (microg/g creatinine) were used as an index of long-term cadmium exposure. Quantitation of cadmium was performed using electrothermal (graphite furnace) atomic absorption spectrometry. The urinary cadmium excreted displayed a normal frequency distribution. The average urinary cadmium level did not exceed the WHO maximum tolerable internal dose for the non-exposed population (2 microg/g creatinine). Body iron stores reflected by serum ferritin levels did not show any correlation with cadmium burden in both males and females, although a relatively stronger influence of body iron store status on cadmium burden was shown in females. When the levels of serum ferritin were stratified into five levels (<20, 20-100, 101-200, 201-300, and >300 microg/l), a significant difference in total cadmium body burden was observed between females and males only in the group with a low level of serum ferritin of <20 microg/l. The cadmium body burden in females was about twice that in males in this group. PMID- 19806463 TI - Anthropogenic influence on surface water quality of the Nhue and Day sub-river systems in Vietnam. AB - In order to investigate the temporal and spatial variations of 14 physical and chemical surface water parameters in the Nhue and Day sub-river systems of Vietnam, surface water samples were taken from 43 sampling sites during the dry and rainy seasons in 2007. The results were statistically examined by Mann Whitney U-test and hierarchical cluster analysis. The results show that water quality of the Day River was significantly improved during the rainy season while this was not the case of the Nhue River. However, the river water did not meet the Vietnamese surface water quality standards for dissolved oxygen (DO), biological oxygen demand (BOD(5)), chemical oxygen demand (COD), nutrients, total coliform, and fecal coliform. This implies that the health of local communities using untreated river water for drinking purposes as well as irrigation of vegetables may be at risk. Forty-three sampling sites were grouped into four main clusters on the basis of water quality characteristics with particular reference to geographic location and land use and revealed the contamination levels from anthropogenic sources. PMID- 19806464 TI - Metastasis-induction and apoptosis-protection by TWIST in gastric cancer cells. AB - TWIST, a basic helix-loop-helix transcription factor, has been recently reported to play an important role in tumorigenesis of human cancer through converting the early stage tumors into invasive malignancies. Upregulation of TWIST is often found in cancer patients, especially those with shorter survival period and poor response to chemotherapy. Here we studied the functions of TWIST on regulating migration rate, apoptosis, and gene expression in gastric cancer cells. TWIST expression is elevated in MGC-803 and HGC-27 cells that exhibit high invasive potential; whereas it is reduced in BGC-823 and SGC-7901 cells that possess relatively low invasive content. To evaluate functional consequences of TWIST induction, we examined the effect of TWIST on cell migration and apoptosis. Overexpression of TWIST in BGC-823 cells resulted in increased migration content and decreased sensitivity to the arsenic oxide-induced cell death. Moreover, small interference RNA-mediated TWIST ablation in MGC-803 and HGC-27 cells showed suppressed migration ability, increased induction of apoptosis in response to arsenic oxide, and elevated cell cycle arrest. Furthermore, we found a negative correlation between the TWIST level and p53 level, probably due to transcriptional regulation. Our results have identified TWIST as a critical regulator of gastric cancer cell proliferation and migration, suggesting a potential therapeutic approach to inhibit the growth and metastasis of gastric cancer through inactivation of TWIST. PMID- 19806465 TI - Association between plasma total cholesterol concentration and incident prostate cancer in the CLUE II cohort. AB - Statin drugs appear to protect against advanced and possibly high-grade prostate cancer, perhaps through cholesterol-lowering. Thus, we evaluated the association between plasma cholesterol and prostate cancer. We conducted a prospective study in the CLUE II cohort of Washington County, MD. Included were 6,816 male county residents aged 35+ years old who did not have a cancer diagnosis at baseline in 1989. Plasma cholesterol, measured enzymatically at baseline, was categorized by clinical cutpoints. Cox proportional hazards regression was used to estimate multivariable-adjusted hazard ratios (HR) and 95% confidence intervals (CI) for total (n = 438) and high-grade (Gleason sum > or =7, n = 137) prostate cancer. Compared to men with high cholesterol (> or =240 mg/dl), men with desirable (<200 mg/dl) or borderline (200 to <240 mg/dl) levels were less likely to develop high grade prostate cancer, particularly when restricting to organ-confined cases (HR: 0.68, 95% CI 0.40-1.18; P trend = 0.12) and among men with higher BMI (HR: 0.36, 95% CI 0.16-0.79; P trend = 0.02). Results were unchanged after excluding cholesterol-lowering drug users. Cholesterol was not associated with total prostate cancer. Our study supports two prior ones suggesting that cholesterol influences risk of high-grade prostate cancer, and indirectly supports the hypothesis that cholesterol-lowering is a mechanism by which statins are protective. PMID- 19806466 TI - Clinical pharmacy services in an Iranian teaching hospital: a descriptive study. AB - OBJECTIVE: To understand the types of services provided by Iranian clinical pharmacists in nephrology and infectious disease wards, the acceptance rate of clinical pharmacy services in these wards by physicians and the clinical significance of these services in the main teaching hospital in Iran. SETTING: Nephrology and infectious disease departments of a university hospital in Iran. METHODS: During a 12-month prospective data gathering phase, details of all clinical pharmacy services in the nephrology and infectious disease wards of a large university hospital were recorded in the pharmacotherapy monitoring forms. Significance impact of clinical pharmacists' services was assessed according to the guidelines of The Society of Hospital Pharmacists of Australia. MAIN OUTCOME MEASURE: Number and type of services provided. RESULTS: During 1 year, clinical pharmacists contributed to 1,386 services for 1,105 patients who were admitted in these two wards; of these services, about 95% were accepted by the physicians and about half of them were of moderate-to-life saving clinical significance. Also at least 32% of services were considered to reduce the cost of drug therapy. CONCLUSIONS: These results support the importance of clinical pharmacists' participation in health care team rounds to improve the overall quality of medication therapy, enhance patient care and outcome and reduce drug costs to patients and society. PMID- 19806467 TI - Nucleotide sequence and expression of the 14-3-3 from the halotolerant alga Dunaliella salina. AB - Previously we reported the nucleotide sequence of a 14-3-3 cDNA cloned from the unicellular green alga Dunaliella salina, however, the nucleotide sequence of this gene have not been reported so far. In the present study, the cloning and characterization of the nucleotide sequence, the gene copy and expression were undertaken. The coding sequence of the gene was found to be interrupted by five introns of 132, 266, 153, 152 and 625 bp, respectively. Introns 3-5 were found in conserved positions as compared to the Chlamydomonas reinhardtii 14-3-3 gene. D. salina 14-3-3 cDNA was inserted into the prokaryotic expression plasmid pET-28 and transformed into E. coli BL21, and the recombinant expressed 14-3-3 protein was purified from E. coli and immunized the rabbit. Indirect ELISA coated with 14 3-3 illustrated that the rabbit antisera titration was 1:1.00E + 06. Western blotting assays confirmed that prepared rabbit antibodies could recognize the recombinant 14-3-3 protein. Southern blotting results showed that there was only one copy of the 14-3-3 present in the genome of D. salina and 14-3-3 expression did not change throughout the Dnualiella cell cycle. PMID- 19806468 TI - Age trajectories of quality of life among older adults: results from the English Longitudinal Study of Ageing. AB - PURPOSE: To explore age-trajectories of quality of life (QoL) and influences on them in a 4-year period among older adults living in England. METHODS: Data come from three waves (2002-2003 and 2006-2007) of the English Longitudinal Study of Ageing, a large panel study of 11,392 individuals aged 50 and over. We used Latent Growth Curve models and ageing-vector graphs to describe both individual differences and average population age-trajectories in QoL (measured by the CASP19 questionnaire). RESULTS: QoL at baseline was poorer for older than younger respondents, with the differences widening with age. QoL also declined more rapidly for older individuals. Gender, education, depression, limiting long standing illness, difficulty with ADL-s, lack of wealth, non-employment, decreased number of friends and low positive support had a negative impact on QoL. Living with a partner had a positive effect on the QoL of men but not of women. The ageing-vector graphs revealed a clear gradient in age-trajectories of QoL for those in the best to the worst psychosocial, socioeconomic and health conditions. CONCLUSIONS: Younger old adults can be prepared for further ageing by increasing their network of friends and engaging with the wider community while they are able. PMID- 19806470 TI - Editor's overview: a special issue on "risk and responsibility". PMID- 19806469 TI - A comparison of two microbial detection methods used in aseptic processing of musculoskeletal allograft tissues. AB - Tissues from 78 musculoskeletal donors were concurrently tested for microorganisms using both a swab and liquid culture method. An aggregate total of 20 organisms were detected by both methods. The swab detected 4/20 organisms while the liquid culture detected 18/20 organisms. The swab method yielded sensitivity and negative predictive values of 20 and 92.3%, respectively. Comparatively, the liquid culture displayed a sensitivity of 90% and a negative predictive value of 99%. These results clearly demonstrate that the liquid culture method is superior to swab cultures in microbial detection. Additional studies are necessary to determine the optimal culture conditions for different types of tissues when utilizing the liquid culture method. PMID- 19806471 TI - Characterization of the shsp genes in Drosophila buzzatii and association between the frequency of Valine mutations in hsp23 and climatic variables along a longitudinal gradient in Australia. AB - The small heat shock gene (shsp) cluster of Drosophila buzzatii was sequenced and the gene order and DNA sequence were compared with those of the shsps in Drosophila melanogaster. The D. buzzatii shsp cluster contains an inversion and a duplication of hsp26. A phylogenetic tree was constructed based on hsp26 genes from several Drosophila species of the Sophophora and Drosophila subgenera. The tree shows first a separation of the Sophophora and the Drosophila subgenera and then the Drosophila subgenus is divided into the Hawaiian Drosophila and the repleta/virilis groups. Only the latter contain a duplicated hsp26. Comparing the gene organisation of the shsp cluster shows that all the Drosophila subgenus species contain the inversion. Putative heat shock elements (HSE) were found in the promoters of all the shsp and putative regulator elements for tissue specific expression were found in the promoter of hsp23, hsp27 and one of the hsp26 genes. hsp23 was found to be polymorphic for four non-synonymous changes that all lead to exchange of a Valine. The duplicated hsp26 gene in D. buzzatii (phsp26) was polymorphic for two non-synonymous changes. The allele frequencies of these variants were determined in nine D. buzzatii populations covering most of its distribution in Australia using high-resolution melting curves. The allele frequencies of one of the hsp23 variants showed a significant linear regression with longitude and the pooled frequency of the four Valine changes of hsp23 in the nine populations showed a significant linear regression with longitude and with a composite measure of climatic variables. PMID- 19806472 TI - High frequency of nonrecurrent MECP2 duplications among Brazilian males with mental retardation. AB - Structural variations that affect the copy number of the MECP2 gene were shown to cause mental retardation in males by driving the overexpression of this gene. To access the impact of these rearrangements in males with unexplained mental retardation, we have performed a quantitative real-time polymerase chain reaction assay using SYBR Green I chemistry to quantify MECP2 gene copy number in 145 Brazilian males with mental retardation of unknown cause. Three patients carrying MECP2 duplications (approximately 2%) were identified. The analysis of additional markers flanking the MECP2 region showed that the duplications observed are nonrecurrent. Expression studies in two of these patients revealed the overexpression of the MECP2 gene compared to the expression level observed in controls. These findings corroborate other recent reports in the literature and highlight that the overexpression of MECP2 caused by duplications involving this gene is a relatively frequent genetic cause of mental retardation in males, highlighting the importance of MECP2 gene dosage for diagnostic purposes in such cases. PMID- 19806474 TI - Ventilator-associated pneumonia in a neurologic intensive care unit does not lead to increased mortality. AB - BACKGROUND: Ventilator-associated pneumonia (VAP) is the most common nosocomial infection among medical intensive care unit (ICU) patients and is associated with increased mortality and length of stay (LOS). Neurologic disease is a risk factor for VAP development, but the relationship between VAP and outcomes in patients admitted to the ICU for neurologic reasons remains largely unknown. METHODS: All mechanically ventilated patients over a 2-year period with neurovascular disease were included in a retrospective study. Data collected included patient demographics, dates of admission and discharge, LOS, and ventilator hours. Comparisons between neurologic patients who did and did not develop VAP were made using univariate and multivariate analysis. RESULTS: Of 585 intubated neurovascular patients, 24 (4.1%) developed VAP. Compared with those who did not develop VAP, those with VAP were younger (51.8 +/- 13.9 years vs. 58.8 +/- 15.9 years, P = 0.03), had increased LOS (32.6 +/- 29.2 days vs. 14.5 +/- 7.8 days, P < 0.001), and more ventilator hours (272 +/- 257 h vs. 85.9 +/- 140 h, P < 0.001). There was no difference in mortality between patients with and without VAP (25.0% vs. 28.3%, P = 0.72). VAP was not an independent predictor of mortality in a multivariate model (OR 1.11, 95% CI 0.37-3.30, P = 0.855). CONCLUSIONS: VAP in neurocritical care patients is associated with increased LOS and ventilator hours, but is not associated with increased mortality, contrary to prior studies in medical ICU patients. PMID- 19806473 TI - Cerebral autoregulation is influenced by carbon dioxide levels in patients with septic shock. AB - BACKGROUND: Altered brain perfusion may play an important role in the development of sepsis-associated encephalopathy. However, whether or not cerebral autoregulation (CA) is preserved in such condition has been debated. CA is dependent on cerebral vascular tone, the main determinant of which is the concentration of carbon dioxide (CO2). The purpose of this study was to evaluate the influence of PaCO2 on the cerebral autoregulatory capacity in patients with septic shock. METHODS: Using transcranial Doppler sonography recordings from the middle cerebral artery (MCA), we evaluated the static cerebral autoregulatory responses within the first 3 days of septic shock. Changes in cerebrovascular resistance (CVR) were calculated from the changes in the mean velocity in the MCA (VMCA, cm/s), in response to an increase in mean arterial pressure (MAP, mmHg) induced by vasopressors. The cerebral autoregulation index (CAI) was calculated as the ratio of the relative changes in CVR and MAP (CAI = DeltaMAP%/DeltaCVR%), with normal values ranging between 0 and 2. RESULTS: We studied 21 mechanically ventilated patients, with a baseline MAP of 65 +/- 6 mmHg, a mean VMCA of 60 +/- 20 cm/s and a median PaCO(2) of 35 [28-49] mmHg. Fourteen of the 21 patients had impaired CA, including 7 of the 14 patients with a PaCO2 <40 mmHg and all 7 patients with a PaCO2 >40 mmHg (Fisher's exact test, P = 0.046). CONCLUSIONS: According to these data, CA is impaired in the majority of patients with septic shock, especially in the presence of hypercapnia. PMID- 19806475 TI - Acute coagulopathy in isolated blunt traumatic brain injury. AB - BACKGROUND: The role of acute coagulopathy after traumatic brain injury (TBI) on outcome has gained increasing appreciation over the recent years. This study was conducted to assess the frequency, outcome, and risk factors associated with this complication. PATIENTS AND METHODS: Using the large, multi-center population based Trauma Registry of the German Society for Trauma Surgery (TR-DGU), we retrospectively analyzed adult patients with isolated blunt TBI (intracranial AIS(HEAD) >or= 3 and extracranial AIS scores <3) for the presence of acute post traumatic coagulopathy upon emergency room (ER) arrival. Coagulopathy was defined as prothrombin time test (Quick's value) <70% and/or platelets < 100,000/microl. RESULTS: From a total of 3,114 eligible patients with isolated TBI, 706 (22.7%) presented with coagulopathy upon ER arrival. Coagulopathy was associated with higher rates of craniotomies (P = 0.02), of single and multiple organ failure and with less intubation-free days. In surviving patients, ICU length of stay and hospital length of stay were significantly longer, if coagulopathy had been present at admission. The overall hospital mortality was 50.4% (n = 356) in patients with coagulopathy vs. 17.3% (n = 417) in non-coagulopathic patients (all P < 0.001). Multivariate analysis identified AIS(HEAD) severity grade, GCS or=2,000 ml and age >or=75 years as independent risk factors for coagulopathy after TBI. Acute coagulopathy in TBI had an adjusted odds ratio for hospital mortality of 2.97 (CI(95): 2.30-3.85; P < 0.001). CONCLUSION: Coagulopathy upon ER admission is frequent after isolated blunt TBI and represents a powerful, independent predictor related to prognosis. Future research should aim to determine the beneficial effects of early treatment of TBI associated coagulopathy. PMID- 19806476 TI - Arginine-vasopressin V1 but not V2 receptor antagonism modulates infarct volume, brain water content, and aquaporin-4 expression following experimental stroke. AB - BACKGROUND: Aquaporin-4 (AQP4) plays an important role in the evolution of ischemia-evoked cerebral edema. Experimental studies have also demonstrated anti edema effects of arginine-vasopressin (AVP) antagonists. In a well-characterized murine model of ischemic stroke, we tested the hypotheses that treatment with selective AVP V(1) but not V(2) receptor antagonist (1) attenuates injury volume and ischemia-evoked cerebral edema; and (2) modulates ischemia-evoked AQP4 expression. METHODS: Isoflurane-anesthetized adult male C57bl/6 mice were subjected to 60 min of middle cerebral artery occlusion (MCAO) by the intraluminal suture technique. Adequacy of MCAO and reperfusion was monitored with laser-Doppler flowmetry over the ipsilateral parietal cortex. Mice were treated with intracerebroventricular injection of selective AVP V(1) and V(2) receptor antagonist or control vehicle (0.9% saline). Infarct volume (tetrazolium staining), cerebral edema (wet-to-dry ratios) and AQP4 protein expression (immunoblotting) were determined in different treatment groups in separate sets of experiments at 24 h of reperfusion. RESULTS: Infarct volume (percentage of contralateral structure; mean +/- SEM) was significantly attenuated in mice treated with 500 ng V(1) receptor antagonist as well as at a dose of 1000 ng compared to controls. However, there was no difference in infarct volume following treatment with 1000 ng V(2) antagonist as compared to controls. Water content in the ischemic hemisphere was significantly attenuated with V(1) receptor antagonist (1000 ng) but not with V(2) receptor antagonist as compared to controls. Treatment with AVP V(1) receptor antagonist (1000 ng) but not V(2) receptor antagonist, significantly upregulated AQP4 protein expression (% beta actin) compared to saline-treated mice in ipsilateral (ischemic) cerebral cortex. CONCLUSIONS: These data demonstrate that following experimental stroke AVP V(1) receptor antagonism: (1) attenuates injury volume and ischemia-evoked cerebral edema; (2) modulates AQP4 expression; and (3) may serve as an important therapeutic target for neuroprotection and ischemia-evoked cerebral edema. PMID- 19806477 TI - The role of integrated (18)F-FDG PET/CT in identification of ectopic ACTH secretion tumors. AB - The role of (18)F-Flurodeoxyglucose positron emission tomography ((18)F-FDG PET) scan in localization of ectopic Cushing's syndrome (EAS) tumor is still controversial. Here, we report on the use of integrated (18)F-FDG PET and computed tomography ((18)F-FDG PET/CT) in localization of EAS tumors in patients with ectopic Cushing's syndrome. Five patients, three men and two women, were reported, whose endocrine investigations and negative pituitary imaging were suggestive of ectopic ACTH secretion. (18)F-FDG PET/CT was performed to identify the source of ACTH secretion. Then the patients were suggested to perform pathologic examination. It turned out that all of these five patients have abnormal markedly intense FDG uptake lesions on (18)F-FDG PET/CT images. Four of them underwent lesion resection, whose plasma ACTH and serum cortisol levels returned to normal after the surgery. Also, they were at last remission from all the symptoms. Pathologic results showed one thymic carcinoid, one pulmonary carcinoid, one thymoma, and one pulmonary carcinoid with upper mediastinum carcinoid. Unfortunately, one patient died due to severe infection and electrolyte disorders. (18)F-FDG PET/CT technology integrates PET and CT imaging in one device so as to increase the accuracy of tumor localization and further improve the prognosis of the patients by curative resection. PMID- 19806478 TI - Stimulation of 11beta-HSD1 expression by IL-1beta via a C/EBP binding site in human fetal lung fibroblasts. AB - Proinflammatory cytokines, just like glucocorticoids (GCs), have been reported to upregulate 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) expression in many cell types. This concerted regulation of 11beta-HSD1 by interleukin-1beta (IL-1beta) and GCs is in marked contrast to their antagonistic effects on inflammation. Further, the molecular mechanisms underlying the induction of 11beta-HSD1 by IL-1beta are not very well understood. In this study, we demonstrated that IL-1beta dramatically stimulated 11beta-HSD1 expression and enzyme activity as well as promoter activity including the -64 bp fragment upstream to the transcription start site in human fetal lung fibroblasts (HFL-1). Nucleotide mutations of the proximal CCAAT box within this region abolished the induction of 11beta-HSD1 promoter activity by IL-1beta. Western blotting analysis demonstrated that IL-1beta induced the expression of C/EBPbeta dramatically while C/EBPalpha was barely detectable in HFL-1 cells. Global inhibition of CCAAT/enhancer-binding proteins (C/EBPs) with transfection of C/EBP-specific dominant-negative expression plasmid (CMV500-A-C/EBP) significantly attenuated the induction of 11beta-HSD1 by IL-1beta, whereas over-expression of C/EBPbeta enhanced the expression of 11beta-HSD1. Chromatin immunoprecipitation assay revealed the recruitment of C/EBPbeta to the promoter region containing the C/EBP binding site. In conclusion, IL-1beta induces the expression of 11beta-HSD1 mRNA in the fetal lung tissue through mechanisms that involve C/EBPbeta binding to the promoter. This impact of IL-1beta on the expression of 11beta-HSD1 in human fetal lung cells may explain the alternate mechanism for the lung maturation that appears to occur when there is a risk of premature delivery of the fetus due to the presence of infection. PMID- 19806479 TI - Serum heart type fatty acid binding protein levels in metabolic syndrome. AB - Heart type fatty acid binding protein (H-FABP) is a major cytoplasmic low molecular weight protein and released into the circulation when the myocardium is injured. Previous studies have demonstrated that H-FABP is closely associated with acute coronary syndrome, hypertrophic and dilated cardiomyopathy, heart failure, stroke, obstructive sleep apnea syndrome, pulmonary embolism. The aim of this study was to investigate serum H-FABP value in the patients with metabolic syndrome (MetS). We measured serum H-FABP levels in 55 consecutive patients with MetS, and 73 age-matched control subjects by using a sandwich enzyme-linked immunosorbent assay. Serum H-FABP levels were significantly higher in patients with MetS than in control subjects 18.37 +/- 13.0 and 7.9 +/- 6.5 ng/ml, respectively, (P < 0.001). Serum H-FABP levels were significantly higher in patients with diabetic MetS than in without diabetic MetS, 24.0 +/- 10.2 and 13.9 +/- 12.6 ng/ml, respectively, (P: 0,003). There were statistically significant differences between patients without diabetic MetS and control subjects, 13.8 +/- 12.6 and 7.9 +/- 6.5 ng/ml, respectively, (P = 0.023). Patients with MetS have an increased risk of death from cardiovascular diseases. H-FABP seems to be a marker that will enable the detection of cardiac injury in the early asymptomatic period in patients with MetS. PMID- 19806480 TI - High-resolution melting analysis of ADAMTS18 methylation levels in gastric, colorectal and pancreatic cancers. AB - A disintegrin and metalloprotease with trombospondin motifs (ADAMTS) is a family of proteins characterized by the presence of a metalloproteinase domain linked to a variety of specialized ancillary domains. ADAMTS18 is a putative tumor suppressive gene related to nasopharyngeal carcinoma. We used high-resolution melting (HRM) analysis to detect the methylation levels of ADAMTS18 gene in 100 gastric cancers, 100 colorectal cancers, 70 pancreatic cancers, and equal number of adjacent normal tissues. The frequency of ADAMTS18 methylation in all three types of cancers was significantly higher than that in normal tissues. Expression levels of ADAMTS18 were inversely correlated with methylation levels. No significant association was found between ADAMTS18 methylation status and TNM staging in the cancers. In summary, epigenetic regulation of ADAMTS18 was associated with carcinogenesis. The application of HRM analysis is a fast and high-throughput way to investigate the epigenetic status of ADAMTS18. PMID- 19806482 TI - Thermal injuries caused by hair straightening devices in children: a significant, but preventable problem. AB - The incidence of contact burns from heated hair straighteners is increasing. The aim of this paper is to assess the safety features of hair straighteners, the mechanism of contact burns and the resultant morbidity in children. This study involves retrospective analyses of patients' case notes and examination of hair straighteners during heating and cooling. Grabbing and stepping onto heated devices were the common mechanisms of injury involving mainly hands and feet. Healing time ranged from 2 to 79 days. Nineteen percent required extensive therapy to maintain good function. Most devices had locking mechanisms to keep the internal surfaces closed; however the external surfaces reached 72.8 degrees C and took 8.6 min to cool below 50 degrees C. Contact with heated hair straighteners causes significant morbidity. The devices, including the external surfaces remain dangerously hot for a prolonged period after being switched off. These findings call for appropriate safety measures to be redressed by manufacturers and awareness amongst users to avoid these preventable injuries. PMID- 19806481 TI - A novel role of cyclinD1 and p16 in clinical pathology and prognosis of childhood medulloblastoma. AB - Medulloblastoma (MB) is the most common malignant brain tumor in children. CyclinD1 is strongly implicated in the control of G1 progression and the phosphorylation of the retinoblastoma protein; p16 could specifically interact with cyclinD1 to inhibit the activity of cyclin-dependent kinase 4. The purpose of this study was to investigate whether quantitative assessment of cyclinD1 and p16 may predict the clinical prognosis in MB. We analyzed the expression of cyclinD1 and p16 antigens in a series of 42 MB with various grades and pathological types by immunohistochemical analysis on paraffin-embedded sections. Then, the correlation of cyclinD1 and p16 expression patterns with clinical pathological features of patients and their prognostic relevance were determined. Compared with the normal cerebellum, expression of cyclinD1 in MB was significantly higher [69.05% (29/42) vs. 9.5% (4/42), P = 0.002], expression of p16 was significantly lower [42.9% (18/42) vs. 85.7% (36/42), P = 0.01]. CyclinD1 expression in MB was up-regulated in metastatic stage (P = 0.01), pathological type (P = 0.02), necrosis (P = 0.009), differentiation level (P = 0.01) as well as with differentiation direction (P = 0.03); p16 expression in MB was down regulated with metastatic stage (P = 0.01), pathology type (P = 0.02), necrosis (P = 0.009) as well as with differentiation level (P = 0.01); negative significant correlation between p16 immunostaining and cyclinD1 was observed in MB (rs = -0.36, P = 0.02). We also found a statistically significant relationship between the percent of cyclinD1-positive cells and p16-positive cells in the tumors and clinical outcome, with higher levels of cyclinD1 and lower levels of p16 correlating with a worse prognosis. CyclinD1 and p16 may be independent biomarkers for clinical prognosis in MB. A combined detection of cyclinD1+/p16- expression may benefit us in prediction of a poor survival of MB. PMID- 19806483 TI - Phonological neighbourhood effects in French spoken-word recognition. AB - According to activation-based models of spoken-word recognition, words with many and high-frequency phonological neighbours are processed more slowly than words with few and low-frequency phonological neighbours. Although considerable empirical support for inhibitory neighbourhood density effects has accumulated, especially in English, little or nothing is known about the effects of neighbourhood frequency and its interaction with neighbourhood density. In this study we examine both effects first separately and then simultaneously in French lexical decision experiments. As in English, we found that words in dense neighbourhoods are recognized more slowly than words in sparse neighbourhoods. Moreover, we showed that words with higher frequency neighbours are processed more slowly than words with no higher frequency neighbours, but only for words occurring in sparse neighbourhoods. Implications of these results for spoken-word recognition models are discussed. PMID- 19806484 TI - Disassortative sexual mixing among migrant populations in The Netherlands: a potential for HIV/STI transmission? AB - To gain insight into the transmission of HIV and sexually transmitted infection (STI) among large migrant groups in The Netherlands, we studied the associations between their demographic and sexual characteristics, in particular condom use, and their sexual mixing patterns with other ethnic groups. In 2002-2005, cross sectional surveys were conducted among migrants from Surinam (Afro- and Hindo-), the Netherlands Antilles, Cape Verde, and Ghana at social venues in three large cities. A questionnaire was administrated and a saliva sample was collected for HIV antibody testing. Of 2105 migrants recruited, 1680 reported sexual contacts, of whom 41% mixed sexually with other ethnicities, including the indigenous Dutch population. Such disassortative mixing was associated with being second generation migrant, having several sexual partners, and having a steady and concurrent casual partner. Less disassortative mixing occurred in participants reporting visiting the country of origin. The association between condom use and sexual mixing differed by gender, with men using condoms inconsistently being most likely to be mixing with the Dutch indigenous population. HIV infection and recent STI treatment were not associated with disassortative mixing. This study shows substantial sexual mixing among migrant groups. Since disassortative mixing is more prevalent in second-generation migrants, it might increase in the upcoming years. The mixing patterns in relation to concurrency and the reported condom use in this study suggest a possibly increased level of HIV/STI transmission not only within migrant groups but also between migrant groups, especially via men who mix with the indigenous population and via migrant women who mix with non-Dutch casual partners. Although the observed HIV prevalence in migrants (0.6%) is probably too low to lead to much HIV transmission between ethnicity groups, targeted prevention measures are needed to prevent transmission of other STI. PMID- 19806485 TI - Associations between substance use, sexual risk taking and HIV treatment adherence among homeless people living with HIV. AB - Prior research suggests that the interconnections between substance use, HIV risk and lack of adherence to HIV medications are especially strong among homeless individuals. Thus, study of these interconnections warrants public health attention. The objectives of this paper are to describe patterns of alcohol and drug use, associations between substance use and participation in high-risk sex, and associations between substance use and adherence to HIV treatment regimens among a sample of 602 homeless or unstably housed HIV-seropositive individuals who are part of a housing-based intervention--the Housing and Health Study. Participants experienced high levels of substance use. Significant associations were found between substance use and adherence to HIV treatment medications, and between substance use and high-risk sexual practices within the entire group. Group analyses by sexual orientation/gender show that the association between substance use and treatment adherence is found primarily among heterosexual males whereas the relationship between several drugs and high-risk sexual practices is strongest among gay and bisexual men. Health professionals working with HIV seropositive individuals should routinely ascertain housing status and screen for substance use and risky sex. PMID- 19806486 TI - Prevalence and predictors of HIV infection amongst Malian students. AB - The objectives of this cross-sectional study were to assess the prevalence and identify predictors of HIV-1 and HIV-2 infection among students in three cities of Mali. Between January and June 2005, we assessed HIV knowledge, attitudes, associated sexual behaviors and tested HIV serostatus among 950 high school and university students in Sikasso, Bamako, and Koulikoro cities, using a combination of enzyme-linked immunosorbent assay (ELISA) and Western blot testing. Univariate and logistic regression analyses were used to determine predictors of infection among students. Mean HIV prevalence was 3.1%, ranging from 1.8% in Sikasso to 3.6% in Bamako. The results showed the presence of all three HIV subtypes in Bamako, though HIV-1 predominated in all cities. Infection rates were slightly higher among males (3.6%) than among females (2.8%), but the difference was not significant. The single significant predictor of HIV infection was knowledge of HIV routes of transmission (p=0.01). HIV prevalence rates observed in this population were higher than general adult prevalence rates previously reported for Mali. HIV/AIDS education and prevention campaigns should be targeted to students at both the secondary and university levels. Students may represent an informative HIV sentinel population for Mali. PMID- 19806487 TI - The impact of incarceration upon adherence to HIV treatment among HIV-positive injection drug users: a qualitative study. AB - INTRODUCTION: HIV-positive injection drug users (IDU) often do not derive the full benefits of highly active antiretroviral therapy (HAART). Among IDU, recent incarceration has been associated with discontinuation of HAART for non-clinical reasons. We sought to qualitatively evaluate experiences with HAART among HIV positive IDU who had been recently incarcerated within provincial prisons in British Columbia in order to identify factors influencing adherence to treatment. METHODS: Twelve in-depth qualitative interviews were conducted with males recruited from a cohort study (ACCESS) involving over 450 HIV-positive IDU. All participants had been incarcerated after initiating HAART. Audio-recorded interviews were conducted to examine experiences of taking HAART in prison, with particular attention to adherence and experiences of treatment discontinuation. FINDINGS: Participant accounts described situations where adherence to HIV treatment was compromised in custody. A small number of participants reported treatment interruptions that lasted over a week when they were unable to obtain HIV medications through institutional healthcare. Short-term interruptions in treatment were said to be common during intake into the correctional system and at the point of release from custody. High levels of HIV discrimination motivate prisoners to hide the fact that they are HIV-positive by making efforts to take medications discreetly, which may result in missed doses. CONCLUSIONS: The current study identified contextual factors within correctional environments that hinder individuals' ability to adhere to HAART. These findings also indicate that improved health services and coordination with community care providers are needed to enhance the quality of HIV treatment within correctional environments. PMID- 19806488 TI - Preventive service needs of young people perinatally infected with HIV in Uganda. AB - The sexual and reproductive health needs of young people perinatally infected with HIV in Uganda remain largely unaddressed by existing HIV/AIDS programs mostly because, such programs encourage young HIV-positive clients to refrain from or postpone sexual activity. This study examines the sexual expressions and experiences as well as the preventive practices of 732 adolescent boys and girls aged 15-19 years who were born with HIV with a view to identifying the preventive service needs of these young people and the implications of these needs for HIV/AIDS programs. The data come from a project on the sexuality of young people perinatally infected with HIV conducted in 2007 in four districts of Uganda, that is, Kampala, Wakiso, Masaka, and Jinja. The analysis involves both quantitative and qualitative approaches: the quantitative approach entails cross-tabulations with chi-square tests as well as significance tests of proportions while the qualitative approach involves an analysis of individual case stories, in-depth probes and focus group discussions for content. The findings show disconnect between: (1) the information the service providers give to the young people and their actual needs and desires; (2) the fears of the adolescents and their actual preventive practices; and (3) the high level of reported condom use and the frequency of use. Programs will therefore need to recognize that young people perinatally infected with HIV are sexually active or anticipate being so in future. Thus, both sexually active and non-sexually active young people require information and services on prevention of unwanted pregnancies as well as avoiding infecting their sexual partners with HIV and re-infecting themselves. Programs will need to devise ways of responding to these needs which should include emphasizing the disclosure of HIV status to the partner as well as the need to accompany such disclosure with consistent condom use. PMID- 19806489 TI - Poverty and psychological health among AIDS-orphaned children in Cape Town, South Africa. AB - This study examined associations between AIDS-orphanhood status, poverty indicators, and psychological problems (depression, anxiety, post-traumatic stress, peer problems, delinquency, conduct problems) among children and adolescents in townships surrounding Cape Town, South Africa. One thousand and twenty-five children and adolescents completed standardized and culturally sensitive cross-sectional surveys. Children orphaned by AIDS had more psychological problems including depression, peer problems, post-traumatic stress, and conduct problems. Specific poverty indicators including food security, access to social welfare grants, employment in the household and access to school were associated with better psychological health. Poverty indicators mediated associations of AIDS-orphanhood with psychological problems. Food security showed the most consistent association with reduced psychological problems. Poverty alleviation measures have the potential to improve psychological health for AIDS-orphaned children in South African townships. PMID- 19806490 TI - Meta-analysis of health and demographic correlates of stigma towards people living with HIV. AB - HIV-related stigma may negatively impact the health, quality of life, social support and well-being of people living with HIV (PLHIV). Previous studies have used diverse samples and a multitude of measurement instruments to examine demographic and health correlates of HIV-related stigma, highlighting the importance of synthesizing findings across different studies to gain a better understanding of these associations. This study examined the relationships between HIV-related stigma and a range of demographic, social, physical and health characteristics. A meta-analysis was conducted to assess the overall strength and direction of these relationships. Twenty-four studies of PLHIV, conducted in North America and published in peer-reviewed journals between January of 2000 and November of 2007, were examined and their findings integrated. The heterogeneity of reported results was also assessed and examined. Our review revealed substantial variability in the ways researchers measure participants' HIV-related stigma as well as their physical, emotional and mental health. In spite of this variability, high stigma level was consistently and significantly associated with low social support (r = -0.369, p<0.0005), poor physical health (r = -0.324, p<0.0005), poor mental health (r = -0.402, p<0.0005), age (-0.066, p<0.05) and income (-0.172, p<0.005). These correlations were of a medium size, which would be recognized by the individual in daily life. Health and mental health professionals working with individuals and families impacted by HIV could benefit from an enhanced understanding of correlates of HIV related stigma, which will inform assessments, interventions and treatment plans. The association between HIV-related stigma and physical health has potential implications for treatment, care and support for people at different stages of HIV infection. AIDS Service Organizations are also encouraged to integrate findings into HIV stigma interventions and social support programs. Additionally, HIV-related stigma scales should be developed and validated, so that future studies using them are able to report findings that are operationally and conceptually consistent. PMID- 19806491 TI - Delayed application of condoms with safer and unsafe sex: factors associated with HIV risk in a community sample of gay and bisexual men. AB - While condom use remains one of the most effective measures to prevent the sexual transmission of HIV, decreasing attention appears to be given to its importance and techniques of effective use relative to potential biomedical technologies. This paper focuses on delayed condom application (DCA), one practice which has been implicated in HIV transmission among gay and bisexual men. It examines the prevalence of the practice within a gay community and explores factors associated with condom use among those who practice only safer sex and those who report at least some unprotected anal sex. Data were taken from an anonymous, cross sectional study of a self-identified sample of gay and bisexual men (N=5080). Among 2614 men who responded to relevant questions, multivariate polytomous logistic regressions were used to identify variables associated with DCA. Nearly, half of the men reported delayed condom application for insertive anal intercourse in the previous 12 months. While the majority of this group also reported episodes of unprotected anal sex, more than 25% of those who reported delayed application only reported safer sexual practices. Most socio-demographic variables found to be associated with unsafe sex in other studies were not associated with DCA. Negative condom use experiences such as tearing, splitting and slippage were associated with delayed application among the two groups. DCA, which may be considered by men as an effective harm reduction strategy requires attention. Interventions to address this behavior need to consider the physical issues of condom use along with the complex array of social, structural, psychological, and interpersonal issues. PMID- 19806492 TI - Intentionality of medication non-adherence among individuals living with HIV/AIDS in Hong Kong. AB - Adherence to antiretroviral therapy is essential to treatment success for individuals living with HIV/AIDS. Despite the wealth of studies in examining antiretroviral non-adherence, few have distinguished between intentional and unintentional non-adherence. The present study attempted to identify factors associated with adherence, intentional non-adherence, and unintentional non adherence among HIV+ individuals using a longitudinal design. Dietary instructions and medication schedule were also included to measure the subtleties of antiretroviral adherence. One hundred and two HIV+ patients who were under antiretroviral therapy were recruited in an outpatient clinic in Hong Kong at baseline with six months follow-up. Using the conventional adherence rate, only 12 (11.8%) of participants reported having missed/ altered medication in the past four days. However, using a more comprehensive assessment, only 27 (26.5%) participants were classified as adherers. Results showed that, adherers were significantly older and had higher adherence self-efficacy than those who were unintentional or intentional non-adherers. Participants classified as unintentional non-adherers had longer length of diagnosis and started medication longer than adherers and intentional non-adherers. Participants classified as intentional non-adherers had worse mental health, higher level of self-stigma, and reported higher score in avoidant coping than adherers and unintentional non adherers. They also scored higher in physical symptoms than adherers. Findings highlight the importance of a reliable, comprehensive measurement for adherence and extend on previous adherence literature that intentional and unintentional non-adherence are separate entities and are associated with different factors. Future research should understand the intentions behind non-adherence and this would serve as an important guide in the development of interventions aimed at improving antiretroviral adherence for HIV+ patients. PMID- 19806493 TI - Pregnancy care in two adolescents perinatally infected with HIV. AB - We describe the main issues encountered in pregnancy care in two perinatally infected adolescents with HIV. Despite the young maternal age, both mothers complied well with visits and treatment during pregnancy and delivered at week 38 through elective caesarean section. Both, however, missed the regular gynaecological and the routine HIV visits scheduled after pregnancy. Both infants following HIV exposure were confirmed HIV-negative at the end of tests performed in the first year of life. A growing number of similar cases is expected as perinatally infected children enter adolescence and become sexually active. These two cases indicate the feasibility of an adequate pregnancy care in very young HIV-positive women, but suggest that potential difficulties may be encountered in this population in maintaining optimal care after delivery. Such objectives might be better obtained with a timely transition of adolescents with HIV from paediatric clinics to a multiservice care setting which includes infectious diseases clinics, obstetric and gynaecologic departments and, particularly, counselling and educational services. PMID- 19806495 TI - Challenges in the evaluation, consent, ethics and history of early clinical trials - Implications of the Tuskegee 'trial' for safer and more ethical clinical trials. PMID- 19806496 TI - Clinical trials for glioblastoma multiforme using adenoviral vectors. AB - Gliomas are among the most lethal malignancies, and constitute more than 70% of all brain tumors. Standard therapy includes surgical resection followed by adjuvant radiotherapy and/or chemotherapy. Malignant gliomas are considered to be non-curable, and the overall prognosis of treatment success is poor with a mean survival of 14.6 months after diagnosis and a 5-year survival rate of 9.8%. The diffusely infiltrating property of the tumors makes total surgical excision often impossible, leading to eventual tumor recurrence. The maximum radiation dose that can be administered to the brain is limited to approximately 60 Gy, which is usually not sufficient to completely eradicate the tumor given that some brain tumors are resistant to radiotherapy. The limitations and short-comings of the available treatment options have provided the impetus to test novel therapy modalities to improve quality of life and increase survival of patients with gliomas. PMID- 19806498 TI - Gene therapy in transplantation: Toward clinical trials. AB - The genetic modification of organs or cells is an attractive approach to protect allogeneic transplants from acute rejection and other complications. The transplant setting offers a unique opportunity to utilize ex vivo gene therapy for the modification of allogeneic organs and tissues prior to implantation. However, significant challenges exist in the application of this concept to human organ transplantation, including the large number of potential molecular targets, the diversity and safety profile of available vector delivery systems and the merging of gene-based therapies with existing immunosuppressive regimens. Accordingly, many different therapeutic concepts and vector systems have been investigated in preclinical studies with the aim of prolonging allograft survival. However, the translation of promising gene therapy strategies to transplant clinical trials has lagged behind the progress made in other medical fields. This review describes the recent preclinical applications of gene transfer to transplantation, and critically evaluates the degree to which gene therapy has been tested clinically in organ transplant recipients. PMID- 19806497 TI - Evading the immune response upon in vivo gene therapy with viral vectors. AB - Gene therapy has the potential to provide minimally invasive and long-term treatment for many inherited disorders that otherwise have poor prognoses and limited treatment options. The sustained therapeutic correction of genetic disease by viral gene transfer has been accomplished in patients with severe immune deficiencies, or by the transduction of an immune privileged site for the treatment of ocular disease. For other diseases and target tissues, immune responses to vectors or transgene products often present major obstacles for therapy. Innate and adaptive immunity, sometimes including pre-existing or memory responses, may contribute by varying degrees to immune-mediated rejection and immunotoxicity. This review provides an overview of the immune responses to in vivo gene transfer with the most commonly used viral gene therapy vectors, and discusses strategies and protocols employed in evading the immune system in order to provide optimal gene therapy. PMID- 19806499 TI - Anatomical and physical barriers to tumor targeting with oncolytic adenoviruses in vivo. AB - Despite promising results in preclinical studies, the therapeutic efficacy of oncolytic adenoviruses (Ads) has not been reproduced in clinical trials in patients with cancer. Recent studies have identified several obstacles that limit the therapeutic efficacy of oncolytic Ads. These obstacles include the lack of a primary Ad attachment receptor on tumor cells in situ, as well as innate host defense mechanisms that recognize Ads as pathogens and act to facilitate their elimination. This review focuses on anatomical barriers inside solid tumors that limit Ad transduction, and specifically barriers associated with several factors: (i) the mechanisms of tumor vascularization and the metabolic microenvironment; (ii) the tumor stroma, which tightly surrounds nests of malignant cells; and (iii) the epithelial phenotype of cancer cells, particularly the trapping of viral receptors in tight and adherens junctions. Potential strategies to overcome these obstacles are also presented. Because published data on the physical barriers to Ad transduction of tumors in situ are limited, studies with other macromolecules used in tumor therapy are also discussed. PMID- 19806500 TI - Adenovirus de-targeting from the liver. AB - Adenovirus (Ad) vectors have substantial potential as biological therapeutics for the treatment of human diseases. Evidence from preclinical studies and clinical trials indicated that several acquired and inherited diseases could be corrected or ameliorated with cell type-specific Ad targeting. One of the major barriers for in vivo Ad targeting is the sequestration of the blood-borne virus in the liver. Significant recent advances have been made in understanding the molecular mechanisms involved in mediating Ad sequestration in the liver. Recognizing the redundancy and synergism between the mechanisms that mediate Ad liver cell transduction and those that mediate the sequestration of blood-borne Ads in the liver creates an opportunity for the development of safe and targeted Ads for gene therapy applications. PMID- 19806501 TI - Reovirus: Rationale and clinical trial update. AB - Early clinical trials of reovirus administered via the intratumoral or intravenous routes in patients with advanced cancers established the safety profile and MTD of these agents. Reovirus is an orphan virus and is the cause of a well-documented subclinical syndrome. Preclinical evidence of the novel mechanisms of anticancer activity of reovirus provided the rationale for advancing this agent into clinical trials. Preclinical studies have also defined the specificity of the antitumor activity of reovirus, and also its efficacy in combination with cytotoxic chemotherapies and immunosuppressants. Early clinical trials of combinations of the injectable oncolytic reovirus therapy Reolysin (Oncolytics Biotech Inc) plus cytotoxic chemotherapies are ongoing, as are phase II trials of Reolysin as a single agent for the treatment of specific tumor types. PMID- 19806502 TI - An analysis of retinal gene therapy clinical trials. AB - In 2008, the initial results from the first three gene therapy trials to use adeno-associated viral vectors to treat an inherited retinal degeneration were published. These trials demonstrated no significant vector-related side effects and provided evidence of successful gene transfer with improved vision in several patients. The success of these trials heralds the beginning of a new era in the treatment of retinal diseases. Much can be learnt by comparing the results of the individual studies, as each has subtle differences, both in surgical technique and vector design. In contrast to laboratory models, humans generally have missense rather than null mutations and are treated later in the disease process than experimental models, when recipient cells are compromised. Intracellular stress responses, such as those regulated by endoplasmic reticulum protein kinase (PERK) and the mTOR pathways, are likely to inhibit the translation of transgenic mRNA by mechanisms that are not evident in null laboratory models treated early in the disease process. Understanding methods to overcome stress responses is likely to be a critical step in translating the applications of gene therapy from animal models to other human retinal diseases. PMID- 19806503 TI - An update on gene therapy in China. AB - Gene therapy has made significant progress in the past decade, and has strong promise for the treatment of chronic diseases. China has emerged as a pioneer in some fields of gene therapy and, in 2003, became the first country to commercialize a gene therapy product, Gendicine. H-101 (Ocorine), the second gene therapy-based medicine, was approved in China in 2005. Commercialization of these two gene therapy products provides China with worldwide recognition in the field of gene therapy. The progress of gene therapy in China has resulted in the establishment of several companies pursuing gene therapy technology for commercial purposes. This review summarizes the recent advances in gene therapy in China, including preclinical studies, clinical trials and product commercialization. PMID- 19806504 TI - Lentiviral vectors in clinical trials: Current status. AB - Lentiviral vectors (LVs) are the most recently developed viral-derived vectors for gene therapy applications, and have demonstrated much promise. The ability to transduce dividing and non-dividing cells, and sustain long-term transgene expression makes LVs uniquely desirable as gene therapy vectors. With advances in vector design and large-scale production, LVs have become safer and more effective gene delivery systems. Since the first clinical trial was approved in 2002, several trials to treat patients with both infectious and genetic diseases have been approved. This review focuses on ongoing and planned trials of LV-based gene therapy. PMID- 19806505 TI - M-01ZH09, an oral live attenuated Salmonella enterica serovar Typhi vaccine for the prevention of typhoid fever. AB - Typhoid fever remains a major health problem globally, particularly in the developing world. The increased emergence of several multidrug-resistant strains of Salmonella enterica serovar Typhi has made the management of the disease increasingly difficult. Although vaccines against typhoid fever are available, improvements are desired in dosage, immunogenicity and tolerability. Emergent BioSolutions Inc is developing M-01ZH09, a single-dose oral vaccine against typhoid fever based on an attenuated strain of S enterica serovar Typhi. Several clinical trials have been completed for the vaccine, including large phase II trials in the US and Vietnam. Additionally, a phase IIc clinical trial was ongoing in India at the time of publication. The available data suggest that M 01ZH09 is well tolerated in clinical trials, and is highly immunogenic, provoking broad immune responses. Because M-01ZH09 involves administration as a single oral dose, the vaccine has the potential to be used in a mass immunization program. PMID- 19806506 TI - VCL-CB01, an injectable bivalent plasmid DNA vaccine for potential protection against CMV disease and infection. AB - Vaccines for the prevention of human CMV (hCMV) infection and disease are a major public health priority. Immunization with DNA vaccines encoding key proteins involved in the immune response to hCMV has emerged as a major focus of hcmv vaccine research. Validation of the protective effect of DNA vaccination in animal models has provided support for clinical trials. VCL-CB01, under development by Vical Inc for the prevention of hCMV infection and disease, is a poloxamer-formulated, bivalent DNA vaccine that contains plasmids encoding hCMV tegument phosphoprotein 65 and the major hCMV surface glycoprotein B. In a phase I trial in healthy adults, VCL-CB01 was well tolerated. In interim results from a phase II trial in hCMV-seropositive hematopoietic cell transplant recipients, VCL CB01 increased T-cell responses compared with placebo. The final results from the phase II trial will be of value for developing strategies to prevent hCMV disease in hCMV-seropositive transplant recipients, and may lead to other trials of VCL CB01 or related vaccines for the prevention of congenital hCMV infection. PMID- 19806507 TI - Albiglutide, an albumin-based fusion of glucagon-like peptide 1 for the potential treatment of type 2 diabetes. AB - Albiglutide, under development by GlaxoSmithKline plc for the treatment of type 2 diabetes mellitus (T2DM), is an albumin-fusion peptide. The compound is a mimetic of glucagon-like peptide 1 (GLP-1), a hormone that decreases glucose levels, but has a short half-life because of degradation by dipeptidyl peptidase (DPP)-4. Albiglutide has a longer half-life as a result of its fusion with albumin and its resistance to degradation by DPP-4, caused by an amino acid substitution (Ala to Glu) at the DPP-4-sensitive hydrolysis site. Data from phase II clinical trials in patients with T2DM revealed that albiglutide was well tolerated and that the drug significantly reduced HbA1c levels compared with placebo. At the time of publication, phase III trials assessing albiglutide alone and in combination with other antidiabetic drugs were recruiting patients with T2DM. Albiglutide represents a promising new drug for the treatment of patients with T2DM; the results of long-term trials are awaited with interest. PMID- 19806509 TI - Fertility preservation: a review of the reproductive strategies. PMID- 19806511 TI - Directional freezing: a solution to the methodological challenges to preserve large organs. AB - Although 60 years have passed since the first successful freezing of cells, whole organ freezing is in its initial phase. Heat and mass transfer has limited the success of large tissue and organ cryopreservation either by slow freezing or vitrification. In this article we discuss the limiting factors for the successful freezing of whole organs, such as homogeneous cooling rate, supercooling, latent heat, and recrystallization. We show how the use of directional freezing technology provides solutions to these problems. Whole ovary cryopreservation and transplantation has been proposed as a new method for preserving long-term ovarian function as opposed to ovarian cortical slices. Fertility preservation will benefit from the success of whole ovary freezing and transplantation. PMID- 19806510 TI - Fertility preservation: definition, history, and prospect. AB - Exposure to gonadotoxic agents for treating cancer and other diseases or postponement of maternity jeopardize the prospects of genetic parenthood and are major factors driving the development of fertility preservation technologies. Although its mainstays, cryopreservation and vitrification, have been adopted from infertility medicine for fertility preservation purposes, there is a need for additional specialized procedures, protocols, and biomedical devices. As its multidisciplinary character and aims diverge compared with other branches of reproductive medicine, we are witnessing the emergence of a field in its own right. PMID- 19806512 TI - The efficacy and safety of human oocyte cryopreservation by slow cooling. AB - Preservation at low temperatures is a practice that has found several applications in medicine and biotechnology. Several types of cells and tissues can be stored virtually indefinitely in liquid nitrogen (-196 degrees C) and returned to physiological temperatures. However, phenomena occurring during cooling and warming may affect the structure and function of the stored material. To some extent, the use of substances with special protective properties, referred to as cryoprotective agents, may avoid this effect, but normally a fraction of the cryopreserved material suffers a partial or total loss of viability. In the context of human in vitro fertilization (IVF), for decades cryopreservation has been used to store different materials of reproductive function, mainly embryos at various developmental stages and spermatozoa. In recent times, the realm of cryopreservation has been expanded to also include fully grown unfertilized oocytes. The storage of oocytes offers obvious advantages, eluding legal and ethical problems that afflict, and sometimes make inapplicable, embryo cryopreservation. Until recently, technical and applicative advances of oocyte cryopreservation were inhibited by the preconception that the unique nature of this cell was an insurmountable obstacle to safe and efficient preservation at low-temperature storage. In fact, through the two alternative methodological approaches of controlled rate slow cooling and vitrification, oocyte cryopreservation has been developed both technically and clinically to a stage that is beginning to challenge the supremacy of embryo freezing as the preferred form of fertility preservation for the treatment of infertile couples. Although several questions remain to be answered as to whether oocyte cryopreservation can ensure adequate standards of efficacy and safety, there appears to be little doubt that in the near future oocyte cryopreservation will be recognized as an established form of IVF treatment. PMID- 19806513 TI - The efficacy and safety of human oocyte vitrification. AB - Vitrification is now a widely applied and highly successful approach for cryopreservation in reproductive biology. Rapidly increasing data prove that it is also a highly efficient technique for low-temperature storage of human oocytes. The latest approaches with appropriately selected cryoprotectants, tools and techniques, and properly adjusted parameters allow close to 100% morphological survival rates, and in vitro embryo development, as well pregnancy and implantation rates, comparable with those achieved with fresh oocytes. With standardization of the technique and elimination of biosafety problems by preserving all the positive features, vitrification may become a common part of the everyday routine in a human embryo laboratory, and it may offer a solution for various medical and social situations as well as for simple logistic problems commonly occurring in assisted reproduction. PMID- 19806514 TI - Fertility preservation with immature and in vitro matured oocytes. AB - The development of an effective oocyte-freezing program will have a major impact on clinical practice in reproductive medicine and will serve as a powerful tool to preserve fertility for teenage girls and young women without male partners or for those individuals who are affected by malignancies. It will also be beneficial to infertile couples who have moral or religious objections about embryo cryopreservation. In addition, a successful oocyte cryopreservation program will eliminate the need for donor and recipient menstrual cycle synchronization and will enable the establishment of oocyte banks, which would facilitate the logistics of coordinating egg donors with recipients. Recent advances in vitrification technology have markedly improved the oocyte survival rate after thawing, and the pregnancy rate is comparable with that achieved with fresh oocytes. However, most studies were performed using in vivo matured oocytes for vitrification. The objective of this article was to review whether immature and in vitro matured human oocytes can be vitrified successfully. The results indicated that although healthy live births can be achieved from the combination of in vitro maturation (IVM) oocytes and vitrification, vitrification of in vitro matured oocytes is less effective than vitrification of in vivo matured oocytes. The results suggest that oocytes should be vitrified at the mature metaphase II stage following IVM rather than at the immature germinal vesicle (GV) stage because the potential of oocyte maturation is reduced by the vitrification of immature oocytes at the GV stage. PMID- 19806515 TI - Fertility preservation: the rationale for cryopreservation of the whole ovary. AB - Recently, much progress has been made in the area of cryopreservation of ovarian tissue, one of the only options for fertility preservation available to women who require immediate gonadotoxic chemotherapy. Human ovarian cortical tissue strips have been cryopreserved, thawed, and autotransplanted with successful reproductive function. Cryopreservation of ovarian cortical strips, however, is limited by the ischemia that occurs at the time of retransplantation. Thus for patients that desire long-term resumption of endocrine function, cryopreservation of the whole ovary with an intact pedicle and vascular supply may be a better option. This article describes recent advances in whole ovary cryopreservation in both animal and human models, with a focus on surgical technique for removal, choice of cryoprotectants, freezing and thawing protocols, and preliminary results with organ retransplantation. Although no human cases of whole ovary retransplantation after cryopreservation have been performed to date, these preliminary studies have been encouraging, and it is likely that this option for fertility preservation will be a viable treatment option in the future. PMID- 19806517 TI - Fresh ovarian tissue and whole ovary transplantation. AB - A series of monozygotic (MZ) twin pairs discordant for premature ovarian failure presented an unusual opportunity to study ovarian transplantation. Ten MZ twin pairs requested ovarian transplantation and nine have undergone transplantation with cryopreservation of spare tissue. Eight had a fresh cortical tissue transplant, one of whom received a second frozen-thawed transplant after the first ceased functioning at 3 years. One had a fresh microvascular transplant. All recipients reinitiated ovulatory menstrual cycles and normal day 3 serum follicle-stimulating hormone levels by 77 to 142 days. Seven have already conceived naturally (three twice). Currently, seven healthy infants have been delivered out of 10 pregnancies. The oldest transplant ceased functioning by 3 years, but then she conceived again after a frozen-thawed secondary transplant. There was no apparent difference in return of ovarian function between the nine fresh ovarian grafts and the one frozen graft. Ovarian transplantation appears to restore ovulatory function robustly. Ten pregnancies, leading to seven healthy infants, including one after cryopreservation, bode well for application to fertility preservation. PMID- 19806516 TI - Frozen-thawed ovarian tissue retransplants. AB - Advances in the diagnosis and treatment of childhood, adolescent, and adult cancer have greatly increased the life expectancy of premenopausal women with cancer. The ovaries are very sensitive to cytotoxic treatment, especially to alkylating agents. The only established method of fertility preservation is embryo cryopreservation according to the Ethics Committee of the American Society for Reproductive Medicine (2005), but this option requires the patient to be of pubertal age, have a partner or use donor sperm, and be able to undergo a cycle of ovarian stimulation, which is not possible when the chemotherapy has to be initiated immediately or when stimulation is contraindicated according to the type of cancer. For patients who need immediate chemotherapy, cryopreservation of ovarian tissue is the only possible alternative. This article reports the techniques and results of orthotopic transplantation of cryopreserved ovarian tissue. Among almost 30 cases reported in the literature, 6 live births have been achieved to date. PMID- 19806518 TI - Fertility preservation for breast cancer patients. AB - Breast cancer is the most common neoplasm in women and accounts for 26% (182,460) of all new cancer cases among women. With the use of screening mammography and advancement in other diagnostic modalities, many cases of breast cancer now can be diagnosed and treated at early stages of the disease. Unfortunately, adjuvant chemotherapy regimens commonly used in the treatment of breast cancer may cause premature ovarian failure due to their cytotoxic effects on the germ cells in the ovary. Therefore preservation of fertility in breast cancer survivors at reproductive age has become an important quality of life issue. Fertility preservation is a recently emerged field of reproductive medicine that may help protect the reproductive capability of the cancer survivors and allow them to have children in the future. Embryo freezing is the most established fertility preservation strategy. But conventional ovarian stimulation protocols are contraindicated in breast cancer patients because of the rise of estrogen and its metabolites to supraphysiological levels. Recently developed ovarian stimulation protocols with aromatase inhibitor letrozole and tamoxifen appear to provide a safe stimulation with endogenous estrogen levels comparable with those achieved in the natural cycle. Oocyte freezing can be considered in single women and in those who do not wish donor sperm. Ovarian tissue freezing could also be an option in breast cancer patients who do not wish or have a time for an in vitro fertilization cycle, which requires 10 to 14 days of ovarian stimulation. PMID- 19806519 TI - [Research in orthopedics and trauma surgery: from where to where?]. PMID- 19806520 TI - [Current application of osteoporosis guidelines--expert recommendations of the German Society of Orthopaedic Surgery and Traumatology]. AB - AIM: As an increasing number of patients suffer from osteoporosis-related disorders worldwide, the medical as well as the socioeconomic impact of this problem is significant. Although evidence-based guidelines for diagnosis and treatment are available, their application in daily practice is insufficient. The aim of our initiative was to develop a strategy for supporting this transfer. METHODS: An expert group of the German Society of Orthopaedics and Traumatology (DGOU) has analysed the current scientific as well as health-care data bases regarding diagnosis and treatment of osteoporosis. Then a set of recommendations has been developed in order to improve this situation. RESULTS: The identified support strategy will focus on better identification of patients with osteoporosis and frailty, enhanced interdisciplinary approaches and increased activity to disseminate available guidelines. Additionally, more research activities are necessary in order to highlight the socioeconomic burden of the disease and to continuously improve surgical treatment strategies in the future. CONCLUSION: To ensure a successful application of the recommendations, continuous support of involved health professionals as well as political institutions, national health insurance systems, scientific societies and patient organisations is necessary. PMID- 19806521 TI - [Clinical and radiological results after distal radius fracture: intramedullary locking nail versus volar locking plate osteosynthesis]. AB - AIM: Treatment of distal radius fractures with the volar locking plate provides a stable fixation and early postoperative function. However, after intramedullary osteosynthesis systems for long bones had been established an intramedullary locking nail (Targon DR, Aesculap) for treatment of distal radius fractures was developed. Thus, the aim of this study was to compare the outcomes after volar locking plate osteosynthesis (Aptus, Medartis) and intramedullary osteosynthesis (Targon DR nail) for the distal radius. METHOD: Between 01/07 and 11/07 13 patients (average age: 60 [30-83] years) with distal radius fractures treated with the Targon DR nail were compared to 12 patients (average age: 57 [20-78] years) treated with a volar locking plate (Aptus, Medartis). Follow-up of the Targon DR group was 5.0 (1.9-8.9) months and 8.5 (2.1-11.6) months for the Aptus group. Parameters were: "Disabilities of the Arm, Shoulder and Hand" (DASH), Gartland-Werley and Martini scores, range of motion (ROM) and radiological parameters. RESULTS: The Targon DR group achieved "good" results in the Gartland Werley score with 6.9 (0-13) points and in the Martini score with 29.9 (20-38) points. A "good" result in the Gartland-Werley score (6.5 [3-11] points) and a "satisfactory" outcome with 27.9 (19-33) points in the Martini score were recorded in the Aptus group. The DASH score showed a higher subjective satisfaction after treatment with the Targon DR nail with 14 (0-39) compared to 23 (4-73) points after treatment with volar plating. ROM in both groups was not significantly different. The Targon DR group achieved an extension/flexion of 99 degrees, an ulnar/radial duction of 50 degrees and a supination/pronation of 139 degrees. After treatment with the volar locking plate the patients showed an extension/flexion of 103 degrees, an ulnar/radial duction of 57 degrees and a supination/pronation of 145 degrees. Postoperative radiographs revealed a better palmar inclination in the Aptus group (-2.5 degrees vs. 5.5 degrees) whereas reconstruction of the radial length was more successful in the Targon DR group (0.8 vs. -0.3 mm). CONCLUSION: The compared osteosynthesis systems achieved equally good functional outcomes after distal radius fractures. However, patients treated with the Targon DR nail showed a superior subjective satisfaction, probably because of the less invasive surgical approach. PMID- 19806522 TI - [Distal intraarticular humerus fracture in the elderly: prosthesis or osteosynthesis?]. AB - AIM: The distal intraarticular fracture of the humerus, even in elderly patients, was treated so far with internal osteosynthesis. Due to the poor bone stock, in association with a complex fracture site, the achieved results may be disappointing. The use of an elbow joint prosthesis may be a solution for these specific problems in elderly patients as long as one takes the features of the prosthesis into account. METHOD: Eleven patients with a mean age of 77 years were followed up for 12 months after implantation of an elbow joint replacement. In the other group we examined 15 patients (average age 73 years) after internal fixation for 20 months. Apart from radiological inspection, we applied the Mayo Elbow Score and documented all complications. RESULTS: There were only type B or C fracture sites in this study. The applied osteosynthesis ranged from the classical bilateral plating with osteotomy of the olecranon to minimal invasive screwing or K-wire pinning with additional postoperative immobilisation. The averaged range of motion amounted to 57 degrees in the osteosynthesis group, compared with 89 degrees in the prosthesis group. In 8 cases we used the semiconstrained Coonrad-Morrey system, and 3 times a hemiprosthetic replacement of the fractured condyles by the Latitude prosthesis. The Mayo score of the group after prosthetic replacement reached 91 compared to merely 77 points in the group after osteosynthesis. After osteosynthesis we saw several major complications, including in 4 cases a partial implant failure with consecutive loss of reposition, 1 case of heterotopic ossification and 1 incomplete sensitive N. ulnaris disorder. CONCLUSION: We recommend osteosynthetic management of type B fractures. The appropriate treatment of C-type fractures remains demanding and leads in cases of reduction malalignment with supportive immobilisation to poor results. Here the primarily implanted elbow prosthesis provides a safe solution for a painfree, stable and mobile joint. PMID- 19806523 TI - [The reversed locked internal plate fixator as an alternative internal fixation of problematic proximal femur fractures]. AB - AIM: The aim of the study was to evaluate the application of a locked internal fixator in complex fractures of the proximal femur, in which the internal fixation with standard implants was not possible due to poor quality of bone or already failed internal fixation in the past. METHOD: Ten patients suffering from a pertrochanteric (n = 5), periprosthetic (n = 1) or subtrochanteric (n = 4) femural fracture between 2003 and 2008 were prospectively registered, underwent open reduction and internal fixation with an "upside-down" femur LISS (less invasive stabilisation system) and were followed up. In all these patients a primary internal fixation had failed or the local bone situation and circulation were poor. The mean follow-up was 14 +/- 25 months. X-ray images and a clinical examination were performed at each appointment. RESULTS: All fractures reached a primarily stable fixation with the locked internal plate fixator. Seven patients showed a complete bone healing after 3 months of follow-up and could bear full body weight afterwards. Three patients with preoperatively existing vascular disease or chronic osteomyelitis showed a deep wound infection postoperatively, which led to the explantation of the implant. CONCLUSIONS: The "reversed" locked internal fixator could be a successful alternative implant for stabilisation of proximal femur fractures which could not be fixated by standard implants due to poor bone quality and circulation. It can also be used as a salvage procedure after internal failed fixation in proximal femur fractures. PMID- 19806524 TI - [Bilateral femoral neck fracture after seizure]. AB - Femoral neck fractures after seizure are rare. This injury can easily be underdiagnosed due to generalised, musculoskeletal pain after seizure. In case of persisting groin pain and limited range of motion X-rays are indicated. Within the first 6 hours after the trauma a joint-preserving therapy is possible. After a delayed diagnosis total hip arthroplasty is necessary. As the result of prolonged intra-articular haematoma the incidence of a femoral head necrosis increases. When choosing the implant, an elevated risk of joint dislocation should be considered. PMID- 19806525 TI - [Single bundle anterior cruciate ligament reconstruction using quadriceps tendon autografts]. AB - AIM: The purpose of this study was to evaluate postoperative outcome and functional scores after single bundle anterior cruciate ligament (ACL) reconstruction with the use of quadriceps tendon autografts after a 16 (12-24) month follow-up. METHOD: 54 patients (9 women, 45 men, body mass index [BMI] 25.3 [18.1-36.3 kg/m (2)]) were included in this prospective series, treated between January 2004 and December 2005. Ligament stability was assessed with the Lachmann and pivot-shift tests and a KT 1000 arthrometer. Tegner index, Lysholm-Gilquist score and the International Knee Documentation Committee (IKDC) were employed to evaluate the functional outcomes. In addition, the single leg hop test was performed. RESULTS: Ligament arthrometry using the KT 1000 arthrometer demonstrated a mean side-to-side difference of 1.53 mm (0.2-4.1 mm). Regarding the Lachmann test, 17 patients (32%) showed 1+ laxity and no patient had an abnormal pivot-shift. The mean Tegner activity score was 4.22 (1-10), the mean Lysholm score was 80.8 (20-100) and the mean IKDC score 68.1 (29-87). Results of the single leg hop test revealed a mean decline of the treated leg to 87.7% (70 100%) of the contralateral side. The mean extension was reduced by 1.7 degrees (0 10 degrees) on the treated knee and the flexion by 2.1 degrees (0-10 degrees), compared to the contralateral knee. CONCLUSION: Single bundle anterior cruciate ligament reconstruction using quadriceps tendon demonstrates highly satisfactory results. Due to a moderate donor site morbidity and preservation of the medial stabilizing structures of the knee a wider use in primary cruciate ligament reconstruction may arise in the future. PMID- 19806526 TI - [Sports after surgical treatment of a herniated lumbar disc: a prospective observational study]. AB - AIM: Lumbar discectomy is the most frequent spinal surgical procedure in Germany. Surgeons vary widely in their preferences with regard to patients' return to sports after spinal surgery. The self-reported sporting activity before and after microdiscectomy was evaluated in a prospective observational study with 5 year follow-up. METHOD: One hundred and five patients (mean age 45.4 years) with self reported sporting activity before lumbar single-level discectomy were re evaluated after 5 years. The clinical outcome was graded using a modified Prolo scale (pain and medication). RESULTS: Based on the Prolo scale, an excellent (n = 80) or good (n = 17) outcome was achieved in 92.4% of patients. Altogether, 91.4% of patients (n = 96) resumed sporting activities: 87 patients returned to their previous sport. Only 8.6% of the patients (n = 9) were unable to return to sports, for various reasons (e.g., poor general health, lumbar pain, or lack of time). The mean time until return to sports after operation was 5.8 months (range 6 weeks to 24 months).The recurrence rate was 5.7% (n = 6), with 3 of the herniations occurring before the return to sporting activities (after 1, 2 and 7 months). CONCLUSIONS: The success rate of single-level microdiscectomy in athletes was frequently good or excellent, with over 90% of patients able to return to sports. The reherniation rate after postoperative athletic activity was comparable with results reported in the literature. PMID- 19806527 TI - [Translaminar fenestration for caudally herniated lumbar discs--a technical note]. AB - Translaminar approaches have been described for lumbar disc herniations which are displaced cranially or laterally into the neuroforamen. This technique provides the advantages of a minimally invasive approach with regard to postoperative instability or partial facet joint resection and avoids a medial arthrectomy. We describe this technique for the first time as an option for a caudally sequestrated disc herniation via a transaxillar sequesterotomy. PMID- 19806529 TI - [Distal radius fractures]. PMID- 19806528 TI - [First description of an intradural disc herniation at the C7-T1 level]. AB - 41 cases of cervical intradural disc herniation have been reported in the literature, none at the C7-T1 level. We describe a case of myelopathy after a fall. The female patient suffered an ataxia of the legs in combination with unilateral C8 cervicocephalgia and hypaesthesia. Magnetic resonance imaging revealed a space-occupying lesion at C7-T1, so we suspected a tumour. After a dorsolateral approach, an intradural disc herniation was found. Six months after the operation, the patient is free of complaints. PMID- 19806530 TI - Risk factors for early and late onset of respiratory symptoms in babies born through meconium. AB - We sought (1) to identify risk factors for meconium aspiration syndrome (MAS) among infants born to women whose labors were complicated by thick-consistency meconium-stained amniotic fluid (MSAF), and (2) to determine whether risk factors and infant prognosis differ according to the time of onset of respiratory distress. We performed a secondary analysis of a multicenter randomized trial of amnioinfusion (AI) for the prevention of MAS among women with thick-consistency MSAF. MAS was defined as onset of respiratory distress requiring oxygen supplementation within the first 4 hours of life. Patients with respiratory symptoms with onset at >or=4 hours were treated as a separate outcome category, "late-onset respiratory distress." We developed peripartum and intrapartum regression models to identify the risk of MAS and its subgroups. A tracing with marked abnormalities was a significant risk factor for moderate/severe MAS, but not for mild MAS or for late-onset respiratory distress. Meconium below the vocal cords and need for resuscitation immediately after birth were risk factors for mild and moderate/severe MAS as well as for late-onset respiratory distress. The risk of nonrespiratory comorbidities varied directly according to the severity of the respiratory distress. Late-onset respiratory distress shares several risk factors with MAS, as defined by Rossi et al. The two conditions may represent different manifestations of the same disease process. PMID- 19806531 TI - Fetal placental thrombosis and neonatal implications. AB - We present the neonatal complications of two premature newborn infants whose placentas demonstrated placental thrombosis in the fetal circulation. Both mothers presented with a 3-day history of decreased fetal movements before delivery. The first infant presented with thrombocytopenia and disseminated intravascular coagulation. The second infant had extended bilateral extended hemorrhagic venous infarctions. Severe fetal placental vascular lesions seem to be a predisposing factor for some adverse neonatal outcomes. We present these two cases with a brief review of the literature. PMID- 19806532 TI - Prediction of diabetes recurrence in women with class A1 (diet-treated) gestational diabetes. AB - We sought to evaluate the likelihood of recurrent diabetes in women with a prior history of diet-treated (class A(1)) gestational diabetes mellitus (GDM). In a retrospective cohort analysis, nulliparous women diagnosed based upon National Diabetes Data Group criteria with diet-treated GDM who had recurrent diabetes in a subsequent pregnancy were compared with those who did not have recurrent diabetes. The probability of recurrent diabetes was calculated using maternal age at first pregnancy, interpregnancy interval, and body mass index (BMI) during the subsequent pregnancy. Three hundred forty-four nulliparous women with diet treated GDM had a subsequent delivery in our database. One hundred thirty-seven (40%) had recurrent diabetes. Women with a history of GDM were more likely to have recurrent diabetes if they were heavier (193 versus 173 lbs; P < 0.001; BMI 35.7 versus 32.2; P < 0.001) and waited longer between pregnancies (2.9 versus 2.4 years, P = 0.02). Age, interpregnancy interval, and BMI can be used to predict diabetes recurrence in pregnant women with a history of GDM. PMID- 19806533 TI - Cutaneous muscle flap: a case report. PMID- 19806534 TI - [Methods for organisational health services research]. AB - On 1 July 2009, the German Network for Health Services Research [Deutsches Netzwerk Versorgungsforschung e. V. (DNVF e. V.)] approved the Memorandum III "Methods for Health Services Research", supported by the member societies mentioned below and published in this journal (Gesundheitswesen 2009; 71: 505 510). The focus of this part of the Memorandum III "Methods for health services research" is on the questions and methods of organisational health services research. In a first step, we describe the central questions which are at the core of organisational health services research. In a second step, we describe the methodological standards and requirements with regard to a) sampling, b) measurement and c) research design. We present a phase model for complex intervention trials. This model allows to conduct high quality organisational health services research, to integrate different methods of social research and to show in which phase they are of special importance. PMID- 19806535 TI - [Assessment of quality of life in health services research - conceptual, methodological and structural prerequisites]. AB - On July 1, 2009, the German Network for Health Services Research [Deutsches Netzwerk Versorgungsforschung e. V. (DNVF e. V.)] approved the Memorandum III "Methods for Health Services Research", supported by the member societies mentioned as authors and published in this Journal [Gesundheitswesen 2009; 71: 505-510]. This is an in-depth publication on "quality-of-life assessment in health services research". Within the context of the health sciences, quality of life (QL) encompasses the subjective well-being and functioning in the physical, psychological and social domains. QL informs about the aspects of health care that "actually get to the patient". QL is what patients primarily experience, what they talk about and what to a large degree affects the acceptance of health care services and processes in the society. Therefore, QL can be considered as a highly important endpoint within health services research. The importance of the construct quality of life is also emphasised in German treaties on social law and utility analyses. This paper is the first account on the relations between health services research and the concept and assessment of QL. Our working group has specified key criteria for QL assessment within studies on health services research. (1) Assessment instruments need to comply with standard quality criteria (reliability, validity, sensitivity, interpretability) and the decision for a particular instrument has to be reasonably justified. (2) Study design and study population have to match with the scientific research question and the sample size has to be biometrically sound. (3) QL assessment including time points over the course of the study has to follow a standardized protocol. (4) Criteria for analysis and interpretation have to be prospectively specified. (5) Studies focusing on diagnostic/therapeutic issues need to specify standards for diagnostic criteria and related therapeutic interventions. PMID- 19806536 TI - [Epidemiological methods for health services research]. AB - On July 1, 2009, the German Network for Health Services Research [Deutsches Netzwerk Versorgungsforschung e. V. (DNVF e. V.)] approved the Memorandum III "Methods for Health Services Research", supported by the member societies mentioned as authors and published in this Journal (Gesundheitswesen 2009; 71: 505-510). This is an in-depth publication on the "epidemiological methods for health services research". Legal, political and economic steps of intervention in the medical care system modify the health services structures and processes but the impact of such interventions on the medical care users has, so far and in general, not been examined scientifically. Due to this lack of evaluation, there is, also with regard to the economic situation within the health system, no transparency of potentially severe effects on healthy and, particularly, on ill people. For this very reason, the main questions and focuses of medical care research deal with prevalence, causes and effects of over, under and inappropriate supply of health services, the interaction between diagnostics and therapy, the processes across different sectors and the complex interdependences of health services. This part of the Memorandum of Deutsches Netzwerk fur Versorgungsforschung e. V. (DNVF e. V., German Network for Health Services Research) will enumerate the methods and instruments that will be used for planned studies and that have been applied for finished studies of health services research and for the evaluation of its quality and value. Health services research takes advantage of the theories and the methods of the disciplines that are involved in its studies. It does not need a specific research methodology; its methods are adapted to the specific research question. It is rather to be expected that certain issues of this research branch and its access to data will lead to the development of new methods. PMID- 19806537 TI - Humanitarian Action Summit: exploring the edges of humanitarian health. PMID- 19806538 TI - Human Resources in Humanitarian Health Working Group Report. AB - Humanitarian responses to conflict and disasters due to natural hazards usually operate in contexts of resource scarcity and unmet demands for healthcare workers. Task shifting is one avenue for delivering needed health care in resource poor settings, and on-the-ground reports indicate that task shifting may be applicable in humanitarian contexts. However, a variety of obstacles currently restrict the ability to employ task shifting in these situations, including issues of regulation, accreditation, funding, and a lack of commonly agreed-upon core competencies for different categories of humanitarian health workers. The Human Resources in Humanitarian Health (HRHH) Working Group during the 2009 Humanitarian Action Summit evaluated the potential strengths and weaknesses of task shifting in humanitarian relief efforts, and proposed a range of strategies to constructively integrate task shifting into humanitarian response. PMID- 19806539 TI - Frequently asked questions of prospective humanitarian health professionals. PMID- 19806540 TI - Civilian protection and humanitarian assistance-report of the 2009 Civilian Protection Working Group. AB - The concept of protecting civilians in armed conflict is enshrined in international humanitarian law and widely acknowledged in humanitarian norms. Making this concept operational in humanitarian service delivery is a challenge. Yet, there are many ways in which humanitarian workers could learn from local people about underlying tensions in their community and with these new insights adjust service delivery accordingly. The 2009 WG developed a qualitative verbal tool that could be used by humanitarian field staff to assist them in understanding issues of civilian protection from the local perspective. The attributes and uses of this tool are described in detail in this WG report. The fundamental aim of this tool is to enhance the capacity of humanitarian workers in their daily work to observe markers of activity and behavior and to inquire and listen from local people about what these markers might or might not mean. The WG emphasized the needs for confidentiality, focus on service delivery (not legal or advocacy work), and iterative routine integration of information gained into field team deliberations. The proposed tool is offered as a possible pilot step towards improving humanitarian understanding and response to local civilian protection needs. PMID- 19806541 TI - Surveillance beyond camp settings in humanitarian emergencies: findings from the Humanitarian Health Information Management Working Group. AB - INTRODUCTION: Surveillance is an essential component of health and nutrition information management during humanitarian situations. Changes in the nature and scope of humanitarian assistance activities have created new challenges in health surveillance, particularly outside of camp-based settings. OBJECTIVES: The primary aim of the Humanitarian Health Information Management Working Group was to identify challenges and areas that need further elucidation in a range of non camp settings, including urban and rural as well as low- and middle-income countries. RESULTS: Three major themes emerged: (1) standardization of measures and methodologies; (2) context in data collection and management; and (3) hidden populations and the purpose of surveillance in urban settings. Innovative examples of data collection and management in community-based surveillance were discussed, including task-shifting, health worker to community member ratio, and literacy needs. CONCLUSIONS: Surveillance in non-camp settings can be informed by surveillance activities in camp-based settings, but requires additional consideration of new methods and population needs to achieve its objectives. PMID- 19806542 TI - Applied technologies in humanitarian assistance: report of the 2009 Applied Technology Working Group. AB - Information and communication technologies, especially in the forms of mobile telecommunications, satellite imaging, and geographical information systems, promise to significantly improve the practice of humanitarian relief. A working group convened at the Humanitarian Action Summit 2009, has begun investigating the challenges to implementing these technologies in field operations, keeping in mind the ethical considerations of linking people to place, and pledging to build a community of practice among academics, practitioners, and developers. PMID- 19806543 TI - The professionalization of humanitarian health assistance: report of a survey on what humanitarian health workers tell us. AB - INTRODUCTION: While the number of humanitarian health workers has grown considerably along with the emphasis on evidence-based humanitarian practice over the last 15 years, no organization exists to ensure ongoing professionalization of this area of expertise. HYPOTHESIS/PROBLEM: To determine whether and to what degree the community of humanitarian health workers self-identify as a professional group; whether a need for a professional society exists to support such a group; and if so, what fundamental elements and activities should it encompass and provide. METHODS: A humanitarian, listserv-based survey was undertaken to evaluate humanitarian professional self-identification, needs for and interest in professional support functions, and priorities toward developing a professional organization to provide needed services. RESULTS: The resulting respondent population represented a broad distribution of age and experience with education and experience being equally important factors in defining humanitarian health professionals. Respondents viewed themselves as humanitarian professionals nearly to the extent they viewed themselves as health-specific technical experts who happen to work in humanitarian assistance; they expressed a strong desire to establish a professional society reflecting that self-identification; and that body should focus on activities of education and training, networking and dialogue, and developing and refining core competencies to support best practices. CONCLUSIONS: Humanitarian health workers self-identify as professionals in humanitarian assistance and as technical experts. A professional organization with specific support functions would be of interest to many humanitarian health professionals. PMID- 19806544 TI - Mental health and psychosocial support in crisis and conflict: report of the Mental Health Working Group. AB - INTRODUCTION: The Working Group on Mental Health and Psychosocial Support was convened as part of the 2009 Harvard Humanitarian Action Summit. The Working Group chose to focus on ethical issues in mental health and psychosocial research and programming in humanitarian settings. The Working Group built on previous work and recommendations, such as the Inter-Agency Standing Committee's Guidelines on Mental Health and Psychosocial Support in Emergency Settings. OBJECTIVES: The objective of this working group was to address one of the factors contributing to the deficiency of research and the need to develop the evidence base on mental health and psychosocial support interventions during complex emergencies by proposing ethical research guidelines. Outcomes research is vital for effective program development in emergency settings, but to date, no comprehensive ethical guidelines exist for guiding such research efforts. METHODS: Working Group members conducted literature reviews which included peer reviewed publications, agency reports, and relevant guidelines on the following topics: general ethical principles in research, cross-cultural issues, research in resource-poor countries, and specific populations such as trauma and torture survivors, refugees, minorities, children and youth, and the mentally ill. Working Group members also shared key points regarding ethical issues encountered in their own research and fieldwork. RESULTS: The group adapted a broad definition of the term "research", which encompasses needs assessments and data gathering, as well as monitoring and evaluation. The guidelines are conceptualized as applying to formal and informal processes of assessment and evaluation in which researchers as well as most service providers engage. The group reached consensus that it would be unethical not to conduct research and evaluate outcomes of mental health and psychosocial interventions in emergency settings, given that there currently is very little good evidence base for such interventions. Overarching themes and issues generated by the group for further study and articulation included: purpose and benefits of research, issues of validity, neutrality, risk, subject selection and participation, confidentiality, consent, and dissemination of results. CONCLUSIONS: The group outlined several key topics and recommendations that address ethical issues in conducting mental health and psychosocial research in humanitarian settings. The group views this set of recommendations as a living document to be further developed and refined based on input from colleagues representing different regions of the globe with an emphasis on input from colleagues from low-resource countries. PMID- 19806546 TI - Health and emergency response: emerging humanitarian challenges. PMID- 19806545 TI - Burden of surgical disease: strategies to manage an existing public health emergency. AB - The World Health Organization estimates that the burden of surgical disease due to war, self-inflicted injuries, and road traffic incidents will rise dramatically by 2020. During the 2009 Harvard Humanitarian Initiative's Humanitarian Action Summit (HHI/HAS),members of the Burden of Surgical Disease Working Group met to review the state of surgical epidemiology, the unmet global surgical need, and the role international organizations play in filling the surgical gap during humanitarian crises, conflict, and war. An outline of the group's findings and recommendations is provided. PMID- 19806547 TI - Impact of the Department of Defense initiatives on humanitarian assistance. PMID- 19806548 TI - How to think about the future: history, climate change, and conflict. PMID- 19806550 TI - The summit. PMID- 19806549 TI - Challenges to the humanitarian community: the role of academia in advancing best practices and policy promotion. PMID- 19806551 TI - Disaster diplomacy: current controversies and future prospects. PMID- 19806552 TI - Health and disaster diplomacy in North Korea: ensuring access and accountability in complex political environment. PMID- 19806553 TI - Evidence-based decision-making (part 1): Origins and evolution in the health sciences. AB - Evidence is defined as data on which a judgment or conclusion may be based. In the early 1990s, medical clinicians pioneered evidence-based decision-making. The discipline emerged as the use of current best evidence in making decisions about the care of individual patients. The practice of evidence-based medicine required the integration of individual clinical expertise with the best available, external clinical evidence from systematic research and the patient's unique values and circumstances. In this context, evidence acquired a hierarchy of strength based upon the method of data acquisition. Subsequently, evidence-based decision-making expanded throughout the allied health field. In public health, and particularly for populations in crisis, three major data-gathering tools now dominate: (1) rapid health assessments; (2) population based surveys; and (3) disease surveillance. Unfortunately, the strength of evidence obtained by these tools is not easily measured by the grading scales of evidence based medicine. This is complicated by the many purposes for which evidence can be applied in public health: strategic decision-making, program implementation, monitoring, and evaluation. Different applications have different requirements for strength of evidence as well as different time frames for decision-making. Given the challenges of integrating data from multiple sources that are collected by different methods, public health experts have defined best available evidence as the use of all available sources used to provide relevant inputs for decision making. PMID- 19806554 TI - Three decades of disasters: a review of disaster-specific literature from 1977 2009. AB - INTRODUCTION: The potential for disasters exists in all communities. To mitigate the potential catastrophes that confront humanity in the new millennium, an evidence-based approach to disaster management is required urgently. This study moves toward such an evidence-based approach by identifying peer-reviewed publications following a range of disasters and events over the past three decades. METHODS: Peer-reviewed, event-specific literature was identified using a comprehensive search of the electronically indexed database, MEDLINE (1956 January 2009). An extended comprehensive search was conducted for one event to compare the event-specific literature indexed in MEDLINE to other electronic databases (EMBASE, CINAHL, AMED, CENTRAL, Psych Info, Maternity and Infant Care, EBM Reviews). RESULTS: Following 25 individual disasters or overwhelming crises, a total of 2,098 peer-reviewed, event-specific publications were published in 789 journals (652 publications following disasters/events caused by natural hazards, 966 following human-made/technological disasters/events, and 480 following conflict/complex humanitarian events).The event with the greatest number of peer reviewed, event-specific publications was the 11 September 2001 terrorist attacks (686 publications). Prehospital and Disaster Medicine published the greatest number of peer-reviewed, event-specific publications (54), followed by Journal of Traumatic Stress (42), Military Medicine (40), and Psychiatric Services (40). The primary topics of event-specific publications were mental health, medical health, and response. When an extended, comprehensive search was conducted for one event, 75% of all peer-reviewed, event-specific publications were indexed in MEDLINE. CONCLUSIONS: A broad range of multi-disciplinary journals publish peer reviewed, event-specific publications. While the majority of peer-reviewed, event-specific literature is indexed in MEDLINE, comprehensive search strategies should include EMBASE to increase yield. PMID- 19806555 TI - Psychosocial health in displaced Iraqi care-seekers in non-governmental organization clinics in Amman, Jordan: an unmet need. AB - INTRODUCTION: Populations displaced by conflict face numerous threats to their psychological well-being; consequently, the prevalence of mental health problems, including anxiety, depression, and post-traumatic stress disorder can be elevated as compared to populations who have not experienced forced displacement. PROBLEM: Little is known about the mental health needs of displaced Iraqis. The factors associated with a need for psychological services among patients at seven clinics served by two NGOs that are known sources of care for the displaced Iraqi population in Amman, Jordan were explored. METHODS: The survey was conducted in January and February 2008 and included a random sample of care seekers from seven clinics selected using interval sampling. Interviews on the health needs of displaced Iraqis and their access to services, including mental health services lasting approximately 20 minutes were conducted. RESULTS: Of the 664 survey participants, 49% (95% CI = 45-53%) of respondents reported needing mental health services and 5% (95% CI = 3-8%) of those in need had access to services. The length of time spent in Jordan (adjusted OR = 1.08; 95% CI = 1.00-1.11) was associated with the need for mental health services and the adjusted odds of requiring psychological services was 39% less for individuals from outside of Baghdad as compared to Baghdad residents (OR = 0.61; 95% CI = 0.38-0.98). Responders citing violence as a factor were twice as likely to be from Baghdad (OR = 2.28; 95% CI = 1.03-6.91), while interviewees reporting displacement as a cause for needing mental health services were twice as likely to be female (OR = 2.14; 95% CI = 1.12-4.18). In individuals 35-44 years of age (OR = 0.36; 95% CI = 0.14-0.87) the need for mental health services due to displacement decreased by 64%, while being a part of a female-headed household decreased the need by 81% (OR = 0.19; 95% CI = 0.06-0.57%). CONCLUSIONS: More attention should be given to expanding the local Jordanian health system capacity for the provision of mental service. Targeted social and psychiatric interventions that are culturally sensitive and aligned with Inter-Agency Standing Committee recommendations should be developed to compliment and expand the existing mental health service capacity in Jordan. PMID- 19806556 TI - If schools are closed, who will watch our kids? Family caregiving and other sources of role conflict among nurses during large-scale outbreaks. AB - OBJECTIVES: The global impact of severe acute respiratory syndrome (SARS) brought attention to the role of healthcare professionals as "first receivers" during infectious disease outbreaks, a collateral aspect to their role as responders. This article records and reports concerns expressed by Canadian emergency and critical care nurses in terms of organizational and social supports required during infectious disease outbreaks. The nature of work-family and family-work conflict perceived and experienced by nurses during infectious disease outbreaks, as well as the supports needed to enable them to balance their social roles during this type of heightened stress, are explored. METHODS: Five focus groups consisting of 100 nurses were conducted using a Structured Interview Matrix facilitation technique. RESULTS: Four emergent themes included: (1) substantial personal/professional dilemmas; (2) assistance with child, elder, and/or pet care; (3) adequate resources and vaccinations to protect families; and (4) appropriate mechanisms to enable two-way communication between employees and their families under conditions of quarantine or long work hours. CONCLUSIONS: Social and organizational supports are critical to help buffer the effects of stress for nurses and assist them in managing difficult role conflicts during infectious disease outbreaks. These supports are necessary to improve response capacity for bio-disasters. PMID- 19806557 TI - Impact of the 2007 Ica earthquake on health facilities and health service provision in southern Peru. AB - INTRODUCTION: Few studies have examined post-disaster coping abilities of health facilities and hospitals. On 15 August 2007, a 7.9 (Richter) earthquake struck off the western coast of Peru near the Department of Ica that devastated the healthcare infrastructure. The impact of the earthquake on the health facilities in the four most affected provinces (Canete, Chincha, Ica, and Pisco), the risk factors associated with the damages incurred, ability to provide services, and humanitarian assistance received were assessed. METHODS: A stratified, systematic sampling design was used to interview a sample of 40 health facilities. The most senior healthcare provider at each facility was interviewed about the facility's experience with service provision following the earthquake. RESULTS: Sixty percent of the facilities reported some damage due to the earthquake; four (10%) were completely destroyed. A total of 78% of the facilities reported providing medical care within the first 48 hours after the earthquake and public facilities were more likely to remain open than were private facilities (p = 0.030). Facilities with an emergency response plan were more likely to provide services than were those without a plan (p = 0.043). Six months after the earthquake, similar numbers of HIV and tuberculosis (TB) patients were receiving treatment at the sampled facilities as prior to the earthquake, and no difficulties were reported in obtaining medications. Some form of assistance, most commonly medications, was received by 60% of the facilities; receipt of assistance was not associated with the facility type, location, damage incurred, or post-disaster service provision. CONCLUSIONS: Response plans were an important factor in the ability to provide services immediately following an earthquake. While a facility's ability to provide services can be determined by structural impacts of the precipitating event, response plans help the staff to continue to provide services and care for affected populations, and appropriately arrange referrals when care cannot be provided at the facility. The findings suggest that health facilities in disasterprone areas should invest in developing and implementing disaster response plans in order to improve health service provision capacity during disasters. PMID- 19806558 TI - Impact of facility damages on hospital capacities for decision support in disaster response planning for an earthquake. AB - Hospitals provide life-saving functions and emergency assistance to communities when disaster strikes. Any damage to hospitals from a disaster, either structural and non-structural, can impair these capabilities. In addition, an inaccurate estimation of the treatment capacities available at hospitals in a disaster affected region can severely affect the success of emergency relief plans. In this paper, the impact of facility damage on hospital operations is estimated using a generic simulation model. From the simulation results, parametric models are developed for estimating hospitals' capacities and patient waiting times that could be used by emergency response teams in making casualty dispatching/routing decisions. PMID- 19806559 TI - Inadequate mass-casualty knowledge base adversely affects treatment decisions by trauma care providers: Survey on hospital response following a terrorist bombing. AB - Healthcare professionals require a unique knowledge base to function effectively during a hospital's response to a mass-casualty incident (MCI). A survey of 128 physicians, nurses, and emergency medical technicians involved in trauma care was conducted to assess their knowledge base and how it affected their decision making in response to a MCI following a terrorist bombing. Three-quarters of the study group responded that = or >20% of the surviving victims were critically injured. Only half of the responders indicated that the main objective of medical management is identifying and treating patients with critical injuries. Forty percent of responders indicated that they would not triage a critically injured victim to immediate care. This survey indicates that further education in the principles of MCI management should be based on critical evaluation of the literature. PMID- 19806560 TI - Management of traumatic liver injuries without a valid trauma system. AB - INTRODUCTION: Despite a global increase in conservative treatment of blunt liver injuries, the number of surgically treated traumas in one major trauma center in Iran has increased. The aim of this study was to unveil the reasons behind this increase in operative management by studying 228 consecutive patients at this regional center. HYPOTHESIS: The increased number of liver injuries operated upon is due to the lack of a solid, well-defined trauma system. METHODS: A retrospective review of all patients admitted for liver trauma at Bahonar Hospital, Kerman, Iran, from March 2001 until March 2006 was conducted. Patient data were collected, studied and statistically processed with regard to demographics, clinical and laboratory findings, surgical procedures, complications, and mortality. RESULTS: All patients who were admitted between 30 360 minutes after injury were included. Twelve hemodynamically stable patients were treated conservatively in an ordinary surgical ward. The remaining 216 patients, 153 of whom had blunt injuries, were hemodynamically unstable. A total of 70 patients were in hemorrhagic shock at the time of arrival. Hemodynamically unstable patients with either the suspicion of associated injuries and/or who displayed clinical deterioration and could not be observed in an ordinary surgical ward were treated surgically. The majority of patients who were operated upon in this series had a grade-II liver injury. The total mortality rate in surgically treated patients was 18.1%. None of the patients treated conservatively died. CONCLUSIONS: Despite the low grade of their liver injuries, the high number of surgically treated patients in this series was due to the absence of a valid trauma system. This result should encourage the authorities to review current trauma systems and trauma surgical guidelines. PMID- 19806561 TI - Driver-related risk factors in commercial motorcycle (okada) crashes in Benin City, Nigeria. AB - INTRODUCTION: There has been global concern regarding road traffic injuries. Motorcyclists constitute a high proportion of fatalities in road traffic crashes. Commercial motorcyclists (Okadas) constitute a unique group in this regard. The purpose of this study was to evaluate driver-related risk factors in Okada accidents in Benin City, Nigeria. METHODS: This was a prospective study. Interviewers administered questionnaires which were used to assess Okada drivers during a two-month period (November-December 2006). RESULTS: A total of 996 Okada drivers were interviewed, 995 males and one female. Their ages ranged from 16-80 years with a mean age of 36.4 +/-2.4 years. In the majority of cases, the maximum educational level achieved was primary or secondary. The majority of Okada drivers (82.8%) took to the Okada business as a last resort. Driver's licenses for Okada operation were possessed by 73.5% of drivers, but only 27.2% had taken a road test before being given a license. No form of training on the use of Okadas was received by 45% of drivers before they commenced operations. Crash helmets were owned by 56.4%, but they did not use them on a regular basis. Inconvenience was the reason provided for poor compliance by 52.7% of drivers. Regular intake of alcohol was present in 39.8% of drivers. CONCLUSIONS: Okada drivers are mainly young males with a low level of education who are ill-prepared and ill-equipped for the road. This is a recipe for traffic crash-related injuries and fatal motorcycle crashes. There is an urgent need for job creation, better licensing procedures, road safety education, national legislation, and enforcement of crash helmet laws as well as alcohol breath tests for Okada drivers in Nigeria. PMID- 19806562 TI - Editorial comments-driver-related risk factors in commercial motorcycle crashes in Benin City, Nigeria. PMID- 19806563 TI - Importance and type of earthquakes and zones. PMID- 19806564 TI - [Analysis of the MUT gene mutations in patients with methylmalonic acidemia]. AB - OBJECTIVE: To investigate the MUT gene mutations in patients with methylmalonic acidemia (MMA), and analyze the genotype-phenotype correlation in patients with methylmalonyl-CoA mutase deficiency. METHODS: The diagnosis of the disease mainly depends on the measurement of C3 (acylcarnitine), C3/C0 (free carnitine) and C3/C2 (acetylcarnitine) in the blood by tandem mass spectrometry, the detection of methylmalonic acid in the urine by gas-chromatography mass spectrometry, the determination of total homocysteine in the serum, and the loading test of vitamin B(12). The entire coding region of the MUT gene was screened by PCR combined with direct DNA sequencing in 21 isolated MMA patients. Novel mutations were identified by restriction fragment length polymorphism (RFLP) and sequence analysis in 100 controls. RESULTS: Seventeen MUT gene mutations were detected in 14 of the 21 patients, among them 8 mutations were novel, and R108H, D244LfsX39 and G544X were more frequent, with the frequencies of 9.5%, 7.1% and 9.5%, respectively. Most mutations were missense mutations (64.7%), and majority of them were in exons 2 and 3 (55.6%). Ten out of the 14 patients with MUT gene mutations had early-onset disease, while one case had late-onset disease, and the remaining 3 cases were detected by newborn screening. In addition, 11 of these 14 patients did not respond to vitamin B(12). CONCLUSION: This study revealed partial MUT gene mutation spectrum in Chinese patients with isolated MMA. The patients carrying MUT mutations often had early-onset disease, and most of them were VitB(12)- non-responsive. PMID- 19806565 TI - [Brain structure abnormality as genetic endophenotype of schizophrenia]. AB - OBJECTIVE: To explore the role of genetic factors in the brain structural variation by using magnetic resonance imaging scan in schizophrenic patients and their unaffected siblings, and to provide experimental evidence for identifying endophenotype of schizophrenia. METHODS: The optimized voxel-based morphometry (OVBM) was used to process the brain magnetic resonance images in 15 first episode drug-naive schizophrenic patients, 19 unaffected siblings of the patients and 38 normal control subjects. The data were analyzed by using general linear model. RESULTS: Compared to the normal control subjects, significant decreases of gray matter was observed in first episode drug-naive schizophrenia in bilateral temporal lobe, bilateral occipital lobe, left insula, left frontal lobe superior frontal gyrus and right lentiform nucleus medial globus pallidus. Significant increases of gray matter in bilateral parietal lobe, bilateral limbic lobe cingulate gyrus in patients group while compared to controls were also found. In unaffected siblings, significant decreases of gray matter was observed in the right temporal lobe, bilateral occipital lobe, left insula, and left frontal lobe precentral gyrus, and significant increases of gray matter were found in left parietal lobe and bilateral cerebellum posterior lobe. Increased gray matter in left parietal lobe precuneus was found in first episode drug-naive schizophrenia when compared with their unaffected siblings. CONCLUSION: There were similar brain structure abnormalities between the first episode drug-naive schizophrenia and their unaffected siblings. Genetic factor may play important role in brain structural abnormality in schizophrenia, which suggested that the brain structural change might be a genetic endophenotype of schizophrenia. PMID- 19806566 TI - [A study on genetic diagnosis for Angelman syndrome]. AB - OBJECTIVE: To study the genetic diagnosis of Angelman syndrome(AS), and provide information for clinic diagnosis and counseling to AS families. METHODS: Methylation specific-PCR (MS-PCR) was used for primary diagnosis of 16 clinically suspected AS cases, and linkage analysis by short tandem repeat (STR) was applied to detect the molecular genetic defect in the nuclear families. RESULTS: In this study, 10 AS patients were identified by MS-PCR, and 9 of them with maternal deletion in chromosome 15q11-q13, 1 with imprinting defect in chromosome 15q11 q13 were confirmed by STR linkage analysis. CONCLUSION: Most of the AS patients could be confirmed by MS-PCR. And STR linkage analysis can detect the molecular defect of AS. It is very important for disease diagnosis, genetic counseling and prenatal diagnosis to perform the related genetic diagnosis. PMID- 19806567 TI - [Linkage analysis of a family with familial hypertriglyceridemia]. AB - OBJECTIVE: To perform linkage analysis and mutation screening in a Chinese family with familial hpertriglyceridemia (FHTG). METHODS: Thirty-two family members including 12 hypertriglyceridemia patients participated in the study. Genotyping and haplotype analysis for 22 subjects were performed using short tandem repeat (STR) microsatellite polymorphism markers on 16 candidate genes and/or loci related to lipid metabolism. Two of the sixteen known candidate genes, APOA2 and USF1 were screened for mutation by direct DNA sequencing. RESULTS: No linkage was found between the candidate genes/loci of APOA5, LIPI, RP1, APOC2, ABC1, LMF1, APOA1-APOC3-APOA4, LPL, APOB, CETP, LCAT, LDLR, APOE and the phenotype in this family. The two-point Lod scores (theta =0) were all less than-1.0 for all the markers tested. Linkage analysis suggested linkage to chromosome 1q23.3-24.2 between the disease phenotype and STR marker D1S194 with a two-point maximum Lod score of 2.44 at theta =0. Fine mapping indicated that the disease gene was localized to a 5.87 cM interval between D1S104 and D1S196. No disease-causing mutation was detected in the APOA2 and USF1 genes. CONCLUSION: The above mentioned candidate genes were excluded as the disease causing genes for this family. The results implied that there might be a novel gene/locus for FHTG on chromosome 1q23.3-1q24.2. PMID- 19806568 TI - [Gene mutation analyses in Chinese children with multiple carboxylase deficiency]. AB - OBJECTIVE: To confirm the diagnosis of multiple carboxylase deficiency (MCD) on the gene level and explore the mutations in Chinese children with MCD. METHODS: Biotinidase (BT) and holocarboxylase synthetase (HLCS) genes were analyzed by PCR and direct sequencing for the 4 BT deficiency patients and 8 HLCS deficiency patients, respectively. The identified mutations in the parents of the patients and 50 normal controls were screened by PCR-restriction fragment length polymorphism and direct DNA sequencing. RESULTS: Total detection rate of gene mutation is 100% in the 12 children with MCD. Six mutations were detected in the 4 children with BT deficiency, they were c. 98-104del7ins3, c. 1369G>A (V457M), c. 1157G>A(W386X), c. 1284C>A(Y428X), c. 1384delA and c. 1493_1494insT. The last four were novel mutations. Four mutations were found in the 8 children with HLCS deficiency. They were c. 126G>T (E42D), c. 1994G>C (R665P), c. 1088T>A (V363D) and c. 1522C>T (R508W). The last two were hot-spot mutations [75%(12/16)], and c. 1994G>C (R665P) was a novel mutation. CONCLUSION: This study confirmed the diagnosis of 12 patients with MCD on the gene level. Six mutations were found in the BT gene and 4 in the HLCS gene, including 5 novel mutations. Two mutations of the HLCS gene are probably hot-spot mutations in Chinese children with HLCS deficiency. PMID- 19806569 TI - [Genetic diagnosis and prenatal diagnosis of Angelman syndrome]. AB - OBJECTIVE: To evaluate the conventional cytogenetic methods in genetic diagnosis and prenatal diagnosis in the family with a proband of Angelman syndrome (AS). METHODS: High-resolution G-banding karyotyping and fluorescence in situ hybridization (FISH) on metaphase chromosomes were performed. RESULTS: Two AS patients and 1 normal fetus in the family were successfully detected by FISH. CONCLUSION: Our result demonstrated that patient with type I AS could be detected by combining the techniques of high-resolution G-banding and FISH with clinical observation, which would offer accurate genetic counseling information to the geneticists and provide the prenatal diagnosis for the AS family. PMID- 19806570 TI - [Two mutations of the KRT6A gene in Chinese patients with pachyonychia congenita type I]. AB - OBJECTIVE: To investigate the gene mutation in a Chinese pedigree and one sporadic case with pachyonychia congenita type I(PC-1), as well as to explore the relationship between the genotype and phenotype. METHODS: The whole coding region of the KRT16 and KRT6A genes were amplified by long-range polymerase chain reaction (PCR). Six patients with PC-1 were studied, five of them were from a pedigree and the other one was sporadic. One unaffected member in the pedigree and 100 unrelated healthy individuals were also studied in order to exclude polymorphism. PCR products were directly sequenced to detect the mutation. RESULTS: No mutations in the KRT16 gene were observed. All patients harbored a mutation in the KRT6A gene. All five patients in the pedigree had a mutation at codon 465 (TAC to CAC) which substitutes tyrosine (Y) by histidine (H). In the sporadic patient, codon 171 (AAC) was mutated to GAC, which changes the asparagines (N) to aspartic acid (D). No such mutations were found in the unaffected member of the pedigree and the 100 unrelated controls. The mutation of Y465H is located at the end of 2B and N171D at the beginning of 1A domain of KRT6A, both are hotspots for pathogenic keratin mutations. CONCLUSION: The mutations Y465H and N171D of the KRT16A gene were detected in the pedigree and the sporadic case respectively. The Y465H mutation was a novel mutation, and the N171D mutation was reported recently. PMID- 19806571 TI - [Rapid detection of the hot spot gene mutations in Chinese patients with nonsyndromic hearing loss by polymerase chain reaction-restrictive fragment length polymorphism]. AB - OBJECTIVE: To develop a rapid genetic diagnosis technique for the patients with hereditary hearing loss by screening hot spots of mutations, namely 235delC of the GJB2 gene, IVS7-2A>G of the SLC26A4 gene, and 1555A>G of mitochondrial 12S rRNA. METHODS: Multiple PCR amplification of the three fragments covering the expected mutations in GJB2, SLC26A4 and 12S were carried out and the amplified products were analyzed by restriction fragment length polymorphism (RFLP). RESULTS: Eighteen homozygous and 18 heterozygous 235delC, 2 homozygous and 13 heterozygous IVS7-2A>G, and 8 homogeneous 1555A>G were detected in the 200 patients with hearing loss. All the results were confirmed by sequencing. The detection rate of the three mutant alleles was 21.7% (71/400 + 8/200 = 0.217) and the genetic diagnosis rate was 14% [(18+2+8)/200 = 0.14]. CONCLUSION: It is a convenient, efficient and economical method to screen the hot spots of mutation in the patient with hereditary hearing loss by using PCR-RFLP. PMID- 19806572 TI - [The effect of silencing Dicer by small interference RNA on the biological characteristics of human glioma cells]. AB - OBJECTIVE: To study the effect of silencing Dicer by small interference RNA (siRNA) to suppress the global microRNA (miRNAs) expression on the biological characteristics of TJ905 glioblastoma cells. METHODS: The silencing effect of RNA interference on Dicer expression was evaluated by reverse transcription polymerase chain reaction (RT-PCR), Western blot analysis and immunofluorescence staining. The cell proliferation rate and cell cycle kinetics were detected by MTT assay and flow cytometry respectively, and the cell invasive ability was evaluated by transwell assay. RESULTS: The siRNA targeting Dicer suppressed the expression of Dicer in TJ905 cells. Meanwhile, the proliferation activity and invasive ability were significantly enhanced in cells transfected with Dicer siRNA compared to those cells transfected with scrambled siRNA and the control cells. CONCLUSION: Suppression of Dicer expression renders the glioma cells harboring more aggressive phenotype. This preliminary finding suggests that global lower expression of miRNAs may play an oncogenic role. PMID- 19806573 TI - [Effect of osteopontin silencing by lentivirus-mediated delivery of siRNA on glioma cell invasion and apoptosis]. AB - OBJECTIVE: To investigate the effect of osteopontin silencing on the invasion and apoptosis of U251 cells. METHODS: The invasion, apoptosis and levels of uPA, MMP 2 and MMP-9 were determined by invasion assay, flow cytometry, Western blot and real-time fluorescence quantitative PCR respectively. RESULTS: Osteopontin small interference RNA (siRNA) inhibited osteopontin expression and cell invasion, promoted apoptosis in U251 cells. In addition, the expression of Bcl-2, uPA, MMP 2 and MMP-9 was decreased, while Bax level was elevated. CONCLUSION: Osteopontin siRNA can inhibit U251 cells invasion via the down-regulation of uPA, MMP-2 and MMP-9 levels, and promote apoptosis through induction of Bax expression and inhibition of Bcl 2 level. It suggests that osteopontin plays an important role in human glioma progression. PMID- 19806574 TI - [Application of fluorescence in situ hybridization in prenatal diagnosis of complex chromosomal abnormalities]. AB - OBJECTIVE: To investigate the application of fluorescence in situ hybridization (FISH) technique in prenatal diagnosis of complex chromosomal abnormalities. METHODS: Eleven prenatal diagnosis cases (8 from amniocentesis and 3 from cord blood) with complex chromosomal abnormalities detected by routine G-banding, were further analyzed by FISH. RESULTS: The FISH technique confirmed the results of balanced chromosome rearrangements detected by G-banding, and clarified the structure of the derivative chromosomes in the 3 amniocentesis samples and the origin of the mark chromosomes in the 2 cord blood samples. CONCLUSION: FISH can be used to diagnose the complex chromosomal abnormalities accurately in prenatal diagnosis, and can provide very useful genetic information for clinical diagnosis and treatment. PMID- 19806575 TI - [Advances in the molecular genetic epidemiology research of age-related macular degeneration]. AB - Age-related macular degeneration (AMD) is the leading cause of blindness in industrialized countries. It is a genetically heterogeneous disorder. With the development of molecular biology and molecular genetics, multiple potentially causative genes have been identified. Current studies of susceptibility genes and genetic epidemiology of AMD are reviewed. PMID- 19806576 TI - [Mutation analysis of a Chinese family with genetic dentinogenesis imperfecta]. AB - OBJECTIVE: To study the genetic etiology of an autosomal dominant dentinogenesis imperfecta in a Chinese family. METHODS: The molecular change of the disease in the family was analyzed through the clinical examination, linkage analysis, mutational screening of the DSPP gene and restriction fragment length polymorphism analysis. RESULTS: The disease related gene was completely linked with microsatellite marker D4S1534. We found a novel mutation in the first exon of the DSPP gene (c.49C>T, p.Pro17Ser). All patients in the family had the mutation, while this mutation was not observed in the normal individuals of this family and 100 unrelated controls. CONCLUSION: The p.Pro17Ser identified in the family was a new pathogenic mutation. Our finding provided further understanding of the molecular mechanism of dentinogenesis imperfecta. PMID- 19806577 TI - [Large germline deletion of the VHL gene in Chinese families with von Hippel Lindau syndrome]. AB - OBJECTIVE: To investigate the large germline deletion of the VHL gene in Chinese families with von Hippel-Lindau disease (VHL). METHODS: The large deletion of the VHL gene in 20 unrelated Chinese VHL families was analyzed by using universal primer quantitative fluorescent multiplex polymerase chain reaction (UPQFM-PCR) and GeneScan analysis. RESULTS: Partial and complete VHL gene deletions were detected in 6 probands, including 3 exon 1 deletions, 1 exon 3 and 2 complete deletions. Of the 2 families with the complete deletions, patients developed multi-centric hemangioblastoma in the retina and central nervous system (CNS), and none developed renal cell carcinoma (RCC). CONCLUSION: Partial and complete VHL gene deletions could be detected in Chinese kindreds with von Hippel-Lindau disease and the test for large deletion of the VHL gene should be implemented in routine DNA diagnosis for VHL disease. Further investigations are required to confirm that entire VHL deletions may be associated with a high risk of hemangioblastomas in the retina and central nervous system. PMID- 19806578 TI - [Mutation analysis of the PAX6 gene in a family with congenital aniridia and cataract]. AB - OBJECTIVE: To identify the mutation in the PAX6 gene in a family with congenital aniridia and cataract. METHODS: Total genomic DNA was extracted from peripheral blood leukocytes of 12 family members including three living affected members and 96 unrelated healthy controls. The coding exons 4-13 of the PAX6 gene with intronic flanking sequences were amplified by polymerase chain reaction (PCR). By comparing sequences of the affected members with that of normal individuals, the disease-causing mutation was detected by direct DNA sequencing. RESULTS: A PAX6 mutation was identified in the 3 patients, which did not exist in the unaffected members and unrelated healthy individuals. The nonsense mutation of C to T was detected at the nucleotide 1143, which converted the Arg codon (CGA) to a stop codon(TGA) (R261X) in exon 10. CONCLUSION: The mutation (R261X) detected in the present study is considered to result in the occurrence of congenital aniridia and cataract in the Chinese family. PMID- 19806579 TI - [R240X mutation of the PAX6 gene in a Chinese family with congenital aniridia]. AB - OBJECTIVE: To study the PAX6 gene mutation in a Chinese pedigree with congenital aniridia. METHODS: Linkage analysis was performed to the Chinese family with congenital aniridia using two microsatellite markers D11S904 and D11S935. Analysis of the PAX6 gene mutation was done by direct sequencing of the whole coding region and exon-intron boundaries of the PAX6 gene in all affected and unaffected individuals in the family. RESULTS: The significant Lod Score of 3.01 was acquired at D11S935. Direct DNA sequence analysis identified a 1080C to T change in exon 9 of the patients, resulting in an Arginine substitution by a stop codon at codon 240 of the PAX6 gene, which was absent in the unaffected individuals in the family and 100 normal controls. CONCLUSION: Our results indicate that mutation p.Arg240Ter of the PAX6 is the genetic basis of the Chinese family with congenital aniridia. PMID- 19806580 TI - [Mitochondrial DNA A1555G mutation of seven families with nonsyndromic hearing loss]. AB - OBJECTIVE: To study mitochondrial DNA (mtDNA) A1555G mutation in seven families with nonsyndromic hearing loss (NSHL). METHODS: Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and real time-amplification refractory mutation system-quantitative PCR (ARMS-qPCR) were applied to detect mtDNA A1555G mutation in seven NSHL families. Related clinical data were also collected and analyzed. RESULTS: The mtDNA A1555G mutation was detected in members from the maternal side, including heteroplasmy and homozygosis, others were negative for this mutation. The copy number of homoplasmic or heteroplasmic mutations of mtDNA A1555G correlated well with the degree of deafness (R = 0.341, P = 0.022 and R = 0.85, P = 0.015, respectively). CONCLUSION: The mutation rate of the mtDNA A1555G is high in the NSHL patients, the mutation type include heteroplasmy and homozygosis. There is significant correlation between the mtDNA A1555G copy number and the severity of hearing loss. PMID- 19806581 TI - [Analysis of -3826A/G polymorphism in the promoter of the uncoupling protein-1 gene in Chinese non-obese and obese populations]. AB - OBJECTIVE: To investigate the -3826A/G polymorphism in the promoter of the uncoupling protein-1 (UCP1) gene and its relations to obesity in Chinese population. METHODS: Three hundred and eighty-four subjects (257 non-obese and 127 obese individuals) from a population of Chinese Han nationality in Chengdu area were studied using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs). Serum lipids were measured by enzymatic kits and apolipoproteins A I, A II, B100, C II, C III and E were measured by the RID kits. RESULTS: The frequencies of A and G alleles at -3826A/G site in obese and non obese groups were 0.508 and 0.492, and 0.467 and 0.533, respectively. It showed no significant difference in allele frequencies between non-obese and obese groups (P > 0.05). In the obese group, subjects with genotype GG had higher serum apo B100 concentrations, and those with genotype AG had higher apo C II and apo C III levels, than those with genotype AA, respectively (P < 0.05). In non-obese male subgroup, subjects with genotype GG had lower serum HDL-C and apo A I levels than those with genotype AA, respectively (P < 0.05), whereas those with genotype AG had lower apo A II levels than those with genotype AA. In addition, in obese males with genotype GG had elevated apo B100 levels compared with those with genotype AA, whereas in obese females with genotype GG had decreased apo AI levels and genotype AG had increased apo C II and apo C III levels compared with those with genotype AG and AA, respectively (P < 0.05). CONCLUSION: -3826A/G polymorphism in the promoter of the uncoupling protein-1 gene was not associated with obesity in Chinese Han population of Chengdu area. It may be associated with serum HDL-C, apo A I and apo B100 levels in non-obese and/or obese subjects of certain genders. PMID- 19806582 TI - [An analysis of the reason for HLA-C allele dropout in five samples by sequence based typing]. AB - OBJECTIVE: To analyze the possible reason for HLA-C allele dropout in routine sequence-based typing (SBT) and improve the accuracy of HLA-C SBT test. METHODS: A total of 620 randomly selected samples from healthy voluntary blood donors in Shenzhen were typed at HLA-C locus by sequence-based typing using the AlleleSEQR HLA-C plus sequence-based typing kit. Samples with no full match result were subjected to cloning and haplotype sequencing of the full-length HLA-C gene. If no novel mutations were found, samples were then retyped, using our self-designed PCR primer pair and PCR conditions replacing the AlleleSEQR HLA-C PCR reagents in the PCR set-up procedure so as to analyze the potential reasons for causing abnormal SBT result. RESULTS: In the 620 samples typed at HLA-C locus using the AlleleSEQR HLA-C SBT commercial kit, 5 samples with no full match result were identified. The closest genotype showed one nucleotide mismatch with many different allele groups at different nucleotide position. Based on the PCR-SBT nucleotide sequence, heterozygous nucleotides were determined only in exon 4, whereas the nucleotides in exon 2 and 3 were all homozygotes. The results showed that HLA-Cw*0706 allele dropout existed in all the 5 samples with abnormal SBT results initially identified by AlleleSEQR HLA-C SBT kit, no novel mutation was found. CONCLUSION: The results indicate that the PCR primer pair incompatible with DNA template may result in allele dropout in HLA-C SBT test. Based on the characterization of HLA-C full-length, it is essential to develop HLA-C SBT kit suitable for Chinese population in the future. PMID- 19806583 TI - [Mutation analysis of ATP13A2 gene in Chinese patients with familial autosomal recessive early-onset parkinsonism]. AB - OBJECTIVE: To investigate the mutation characteristics of ATP13A2 gene in Chinese patients with familial autosomal recessive early-onset parkinsonism (AREP). METHODS: Mutations of ATP13A2 gene were screened by polymerase chain reaction combined with DNA direct sequencing in patients with familial AREP. RESULTS: No pathogenic mutations in ATP13A2 gene were detected in this group. Six reported polymorphisms were identified. They were IVS6+70A>G, IVS12+66A>G, m.1849C>T, IVS20-56 G>A, m2671C>T and m2824G>A. CONCLUSION: ATP13A2 gene mutations may be rare in Chinese patients with familial autosomal recessive early-onset parkinsonism. PMID- 19806584 TI - [Association between the -1031T/C polymorphism in tumor necrosis factor-alpha gene and unstable angina]. AB - OBJECTIVE: To investigate the association between the -1031T/C polymorphism in the promoter of tumor necrosis factor-alpha (TNF- alpha) gene and unstable angina in Chinese Han population. METHODS: The genotype of -1031T/C locus was analyzed by MALDI-TOF in 299 patients with unstable angina and 202 healthy controls. The serum TNF-alpha level was measured by enzyme-linked immunosorbent assay(ELISA). RESULTS: There was no significant difference in the genotype distribution and allele frequencies of the -1031T/C locus between the two groups (P > 0.05). However, stratification by gender showed that the genotype distribution of this locus was obviously different between the two groups in men (P was 0.032). The risk of developing unstable angina in men carrying the CC+TC genotypes were 1.66 fold higher than that in men of TT genotype (95% CI: 1.040 to 2.659). There was no significant difference in the frequencies of the C and T alleles between the two subgroups (P > 0.05). Furthermore, serum TNF-alpha levels of the patients were significantly higher than those of controls (P = 0.028, P = 0.013 in men), but there was no significant difference in the TNF-alpha level among different genotypes. CONCLUSION: The -1031T/C polymorphism of the TNF-alpha gene might be associated with unstable angina in male Han population, especially the C allele carriers might be more likely to be affected by unstable angina than the rest of the population. PMID- 19806586 TI - Retraction: Clinical features study of a brachydactyly type A1 family and the disease-causing gene location. PMID- 19806585 TI - [Association of 5-hydroxytryptamine transporter gene polymorphism with asthma and comorbid depression]. AB - OBJECTIVE: To explore the molecular genetic mechanism of complicating depression in asthma by detecting two gene polymorphisms of 5-hydroxytryptamine transporter (5-HTTLPR/Stin2) gene. METHODS: One hundred fifty-six adults with asthma were collected, and divided into group of asthma with depression (HAMD score > or = 8) and group of asthma without depression or single asthma (HAMD score <8) according to the score of Hamilton depression scale (HAMD). A total of 508 adults with depression alone and 433 healthy individuals were enrolled as controls. The target gene fragments containing the polymorphic regions of 5-HTTLPR and Stin2 were amplified by polymerase chain reaction (PCR). The amplified fragments were then analyzed using agarose gel electrophoresis (AGE) and motored molecular imaging system. RESULTS: The frequencies of genotype and allele distribution of the Stin2 polymorphism showed that males with genotype Stin2.12/Stin2.10 and allele Stin2.10 had higher risk for asthma than the others (Stin2.12/Stin2.10: OR = 2.291, 95% CI: 1.195 and 4.390; Stin2.10: OR = 1.942, 95% CI: 1.069-3.527). No significant difference was found in the frequencies of genotype and allele distribution of the 5-HTTLPR locus between the asthma and healthy control groups and the two stratified by gender. CONCLUSION: The Stin2 polymorphism may play a role in the onset of male asthma. There might be association between the genetic pathogenesis of asthma and depression. PMID- 19806587 TI - [Genetic polymorphisms of 9 non-combined of DNA index system short tandem repeat loci in Guangdong Han population]. AB - OBJECTIVE: To investigate the genetic polymorphisms and their forensic application of 9 non-combined of DNA index system (CODIS) short tandem repeat(STR) loci in Guangdong Han population. METHODS: DNA samples from 500 unrelated individuals were extracted and amplified with fluorescence labeled multiplex PCR system. PCR products were separated and genotyped with capillary electrophoresis. RESULTS: One hundred and fifteen alleles and 160 genotypes were observed in the 9 STR loci, respectively. The heterozygosity was 0.824-0.884, the discrimination power (DP) was 0.925-0.969 and the polymorphism information content (PIC) was 0.77-0.86, respectively. The distribution met the Hardy Weinberg equilibrium (P > 0.05). The total discrimination power was 1.00 x 10( 13), the combined probability of exclusion for trio-paternity testing was 0.999989488. The combined probability of exclusion for duo-paternity testing was 0.873436. CONCLUSION: The 9 STR loci are powerful and reliable for personal identification and paternity testing. They can be used as supplementary loci in fatherless (motherless) testing or cases with mutation events. PMID- 19806588 TI - [Genetic polymorphisms of six X-short tandem repeats loci in Chinese Korean]. AB - OBJECTIVE: To investigate the genetic polymorphisms of DXS7132, DXS6854, DXS6797, DXS9898, DXS8378 and GATA31E08 short tandem repeats (STRs) loci in Chinese Korean ethnic group of Yanbian, Jilin, and to construct a preliminary database. METHODS: The allele frequencies of the six STRs loci in Chinese Korean ethnic individuals were analyzed by multiplex polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis (PAGE). RESULTS: Total of 8, 6, 8, 8, 5 and 10 alleles were observed in each locus respectively. All loci (in female) met Hardy-Weinberg equilibrium (P > 0.05). The statistical analysis of the 6 STR loci showed the heterozygosities were more than 0.4660, the polymorphic information contents (PIC) were more than 0.5293, the haplotype diversity were more than 0.9993, power of discrimination (PD) in females and males were more than 0.7737 and 0.6107, respectively. CONCLUSION: The results showed that, all the 6 STR loci in this study were found to have high heterozygosity and polymorphic information content, so they could provide useful markers for genetic purposes. These results could serve as valuable data to enrich the Chinese Korean ethnic group genetic database and play an important role in genetic study of Chinese population. PMID- 19806589 TI - Histochemical methods for the diagnosis of mitochondrial diseases. AB - Through the process of oxidative phosphorylation (OXPHOS), mitochondria provide cells with required energy in the form of ATP. The organelle possesses its own genome (mtDNA), which encodes for part of the components needed (37 genes encoding either OXPHOS structural subunits or tRNAs and rRNAs). Nonetheless, the majority of structural OXPHOS components (as well as accessory proteins and proteins required for maintenance, replication, and expression of the mtDNA) are encoded by nuclear genes. Due to the dual genetic control and the large number of proteins involved, biogenesis and assembly of the OXPHOS system is complicated, and identifying a specific gene defect can be a difficult and time consuming task. This unit describes procedures for obtaining tissue sections and cell material suitable for histological evaluation of OXPHOS activity and integrity and immunodetection of the complexes in tissue from patients suspected of mitochondrial disease. Emphasis lies on the diagnostic potential of these techniques to differentiate mtDNA from nuclear mutations. PMID- 19806590 TI - Evaluation of the mitochondrial respiratory chain and oxidative phosphorylation system using polarography and spectrophotometric enzyme assays. AB - The oxidative phosphorylation (OXPHOS) system consists of five multimeric complexes embedded in the mitochondrial inner membrane. They work in concert to drive the aerobic synthesis of ATP. Mitochondrial and nuclear DNA mutations affecting the accumulation and function of these enzymes are the most common cause of mitochondrial diseases and have also been associated with neurodegeneration and aging. For this reason, several approaches for the assessment of the OXPHOS system enzymes have been developed. Based on the methods described elsewhere, the assays describe methods that form a biochemical characterization of the OXPHOS system in cells and mitochondria isolated from cultured cells or tissues. PMID- 19806591 TI - Evaluation of the mitochondrial respiratory chain and oxidative phosphorylation system using blue native gel electrophoresis. AB - The oxidative phosphorylation (OXPHOS) system consists of five multimeric complexes embedded in the mitochondrial inner membrane. They work in concert to drive the aerobic synthesis of ATP. Mitochondrial and nuclear DNA mutations affecting the accumulation and function of these enzymes are the most common cause of mitochondrial diseases and have also been associated with neurodegeneration and aging. For this reason, several approaches for the assessment of the OXPHOS system enzymes have been progressively developed. Based on methods described elsewhere, the use of blue native gel electrophoresis (BNGE) techniques to routinely assess the OXPHOS system and screen for enzymatic defects in homogenates or mitochondrial preparations from tissues or cultured cells is described here. PMID- 19806592 TI - Evaluation of the mitochondrial respiratory chain and oxidative phosphorylation system using yeast models of OXPHOS deficiencies. AB - The oxidative phosphorylation (OXPHOS) system consists of five multimeric complexes embedded in the mitochondrial inner membrane. They work in concert to drive the aerobic synthesis of ATP. Mitochondrial and nuclear DNA mutations affecting the accumulation and function of these enzymes are the most common cause of mitochondrial diseases and have also been associated with neurodegeneration and aging. Several approaches for the assessment of the OXPHOS system enzymes have been developed. Based on the methods described elsewhere, this unit describes the creation and study of yeast models of mitochondrial OXPHOS deficiencies. PMID- 19806594 TI - New physician practice certification program. PMID- 19806593 TI - Molecular analysis of Fragile X syndrome. AB - The gene responsible for Fragile X syndrome, fragile X mental retardation-1 (FMR1), contains an unstable sequence of CGG trinucleotide repeats in its promoter region. Expansions of >200 trinucleotide repeats are considered full mutations and typically lead to abnormal methylation of the region resulting in loss of FMR1 expression. Males with loss of FMR1 protein are expected to be affected by Fragile X syndrome while females may or may not clinically manifest features of the condition. The protocols in this unit outline the complementary use of polymerase chain reaction (PCR) and methylation-sensitive Southern blot hybridization to accurately measure trinucleotide repeat size and methylation status. These protocols are also used to evaluate CGG repeat size in two adult onset conditions known for their association with FMR1 premutation alleles, Fragile X Tremor/Ataxia (FXTAS) syndrome and Premature Ovarian Failure (POF). PMID- 19806595 TI - Many unknowns in low-risk prostate cancer treatment. Ongoing studies and biomarker research may shed light on best approach. PMID- 19806597 TI - New technology may determine whether chemotherapy is working after a single cycle. PMID- 19806596 TI - Clinical trials of note. Sorafenib as adjuvant treatment in the prevention of disease recurrence in patients with hepatocellular carcinoma (HCC) (STORM). PMID- 19806598 TI - Body dissatisfaction across cultures: findings and research problems. AB - This review had two aims. The first aim was to describe the existing literature regarding body dissatisfaction across cultures. Previous studies were organized and presented according to three general patterns found in the existing literature, patterns that were to some extent based on the differing degrees of affluence and type of lifestyles in the cultures examined. The review's second aim was to illuminate some of the specific problems that studies of body dissatisfaction across cultures deal with and to provide suggestions for improvement. The suggestions concern how future research may achieve a more inclusive and detailed picture of body dissatisfaction across cultures, for instance by using diversified measurements, providing detailed information about the cultural groups that are examined, investigating how different risk factors may influence the level of body dissatisfaction in different cultures and widening the research geographically. PMID- 19806599 TI - Effect of the adapted Virtual Reality cognitive training program among Chinese older adults with chronic schizophrenia: a pilot study. AB - OBJECTIVE: To examine the effect of the adapted virtual reality cognitive training program in older adults with chronic schizophrenia. METHODS: Older adults with chronic schizophrenia were recruited from a long-stay care setting and were randomly assigned into intervention (n = 12) and control group (n = 15). The intervention group received 10-session of VR program that consisted of 2 VR activities using IREX. The control group attended the usual programs in the setting. RESULTS: After the 10-session intervention, older adults with chronic schizophrenia preformed significantly better than control in overall cognitive function (p .000), and in two cognitive subscales: repetition (p .001) and memory (p .040). These participants engaged in the VR activities volitionally. No problem of cybersickness was observed. CONCLUSIONS: The results of the current study indicate that engaging in the adapted virtual reality cognitive training program offers the potential for significant gains in cognitive function of the older adults with chronic schizophrenia. PMID- 19806600 TI - Cognitive and daily functioning in older adults with vegetative symptoms of depression. AB - OBJECTIVES: In primary care 50-95% of patients with depression present with vegetative symptoms (VS). Based on the extant literature, older adults showing VS (but no dysphoria) may show functional impairment but this hypothesis has not been empirically tested. The goal of this study was to examine neurocognitive and daily functioning of elderly patients showing exclusively VS in comparison with patients presenting with VS and dysphoria. METHODS: Seven hundred and eighty seven primary care patients received measures of neurocognition and daily functioning. Neurocognition was measured with the repeatable battery for the assessment of neuropsychological status (RBANS). Three groups were compared: (1) patients with two or more VS of depression without dysphoria (VS - D), (2) patients with at least one VS and dysphoria (VS + D), and (3) comparison patients without multiple VS or dysphoria. RESULTS: Nearly one third of the sample (31%) fell into the VS - D group, whereas 15% fell into the VS + D group. Both VS groups showed poorer neurocognition and activities of daily living than comparisons. Only one subtest of the RBANS (i.e., picture naming) showed a significant difference between VS + D and VS - D, and there was no significant difference on daily functioning. VS - D patients reported less frequent past history of depression and endorsed less anxiety compared to VS + D. CONCLUSIONS: Elderly patients presenting with clusters of VS with or without dysphoria show poorer neurocognitive and functional performance. Relative poorer cognition and daily functioning in VS - D are potential harbingers of further decline and consistent with under-reporting of sadness in older age. PMID- 19806601 TI - Depression, cognitive reserve and memory performance in older adults. AB - OBJECTIVES: The purpose of this research study was to examine the relationship between education and leisure, as markers of cognitive reserve, depressive symptoms and memory performance in a sample of cognitively normal Irish older adults. METHODS: A cross-sectional survey style design was employed to gather data. A sample of 121 older adults in the Cork area was recruited through publicly advertising for volunteers. Only those volunteers who obtained a score of greater than 23 on the MMSE, and were not taking antidepressant or anxiolytic medications, were included. Data from 99 participants were included in the analysis. RESULTS: Controlling for age and gender, depressive symptoms were found to be associated with poorer immediate recall performance, while greater than 12 years of education was positively associated with delayed recall and savings. Leisure did not emerge as being associated with any of the dimensions of memory assessed. CONCLUSIONS: Depressive symptoms emerged as associated with immediate recall, even though few of the participants met the cut-off for caseness. This may indicate a need for intervention in cases of subclinical depression with associated memory complaints. The association between education level and both delayed recall and savings provides support for the cognitive reserve hypothesis, and may suggest useful non-pharmacological approaches to memory deficits in later life. PMID- 19806602 TI - Robotic-assisted single-incision right colectomy: early experience. AB - BACKGROUND: Application of laparoendoscopic single-site surgery (LESS) is increasing across surgical disciplines. In addition to the possibility of decreased postoperative pain, LESS offers better cosmesis with virtually 'scarless' surgeries, while avoiding the increased costs and complexity of natural orifice surgery. Instrument conflict minimization often requires the crossing of articulating instruments, which we believe can be more intuitively facilitated using the daVinci-S((R)) robotic system. We describe our early experience with three robotic single-incision right hemicolectomies. METHODS: Three robotic single-incision right hemicolectomies were performed using the daVinci-S robotic system, utilizing a single 4 cm incision through or around the umbilicus. The procedure was performed using three robotic arms, a 12 mm camera and two 8 mm robotic ports. A medial to lateral approach was used and an extracorporeal resection and anastomosis was performed after undocking the robot. RESULTS: There were no intraoperative or postoperative complications. Average operative time was 152 min. The first case was converted to non-robotic single incision right hemicolectomy during mobilization of the ascending colon, due to uncontrollable air leakage around the ports. The second and third cases were successfully completed without air loss by purse-stringing sutures around each individual port and the use of the SILS() port, respectively. CONCLUSIONS: Robotic-assisted single-incision right hemicolectomy can be successfully and safely performed using the daVinci-S robotic system. Several techniques may be employed to prevent the loss of pneumoperitoneum. We believe right hemicolectomy lends itself to single-site surgery because specimen extraction requires a 4 cm incision and may confer patient benefit, with decreased postoperative pain and improved cosmesis. By crossing the robotic instruments and reassigning control of the arms, the robot represents a means to help perform these procedures safely by allowing them to be performed in a more intuitive fashion. PMID- 19806603 TI - Depression in Dutch homes for the elderly: under-diagnosis in demented residents? AB - OBJECTIVE: Although community-based studies reported an increased incidence of depression among demented persons compared with non-demented persons, it is not clear whether this relationship also exists among institutionalised elderly persons. The aim of this study was to compare the prevalence of diagnosed depressive disorders and mood symptoms between demented and non-demented residents living in Dutch homes for the elderly. METHODS: Cross-sectional analysis in 16 homes for the elderly of routine outcome measurements by trained nurse assistants using the Resident Assessment Instrument (RAI) between January 2007 and April 2008. Nurse assistants recorded all known medical diagnoses including dementia and depression, as well as a structured observation of the presence or absence of 11 mood symptoms over the last 3 days. RESULTS: 313 demented and 463 non-demented residents with complete data were included (99% of all residents, mean age 84 years). 24.6% of participants were diagnosed with a depressive disorder, with no statistically significant difference between demented and non-demented persons (p = 0.237). Mood symptoms were more prevalent in demented residents (p < 0.001, OR 2.14, 95%CI 1.56-2.93). Among residents with mood symptoms, demented residents were less likely to be diagnosed with a depressive disorder than non-demented residents (p = 0.039, OR 0.61, 95%CI 0.38 0.98). CONCLUSIONS: The prevalence of diagnosed depressive disorders was comparable between demented and non-demented residents. However, demented residents suffered more from mood symptoms and may be at risk of under-diagnosis of depression. PMID- 19806604 TI - Diabetes incidence associated with depression and antidepressants in the Melbourne Longitudinal Studies on Healthy Ageing (MELSHA). AB - OBJECTIVE: Diabetes may be associated with depression and antidepressant medication (ADM) use, but published findings remain equivocal. The authors' aimed to determine the risk of diabetes incidence associated with baseline depression exposures (symptoms and/or ADM use). METHODS: A prospective cohort study was conducted in a regionally representative sample of non-institutionalised older Australian people (N = 1000, aged 65 + year), who were followed up biennially between 1994 and 2004 (attrition was approximately 24%). Analyses excluded participants for prevalent diabetes at baseline, determined by self-report or specific medications. Diabetes incidence was ascertained by first self-report at any follow-up wave. Depression exposures (baseline predictors) were defined by the Psychogeriatric Assessment Scales (PAS) depression scale and ADM use, and classified as: (1) 'symptomatic' (PAS score 5+); (2) 'ADM use'; (3) 'symptomatic or ADM use'; (4) 'symptomatic and no ADM use'; (5) 'asymptomatic (PAS score <5) and ADM use' and (6) 'symptomatic and ADM use'. Covariates were demographic, lifestyle, functional health and chronic disease factors. Cox regressions were used to determined hazard ratios with 95% confidence intervals (HR [95% CI]) for diabetes incidence according to depression exposures, adjusted for significant covariates. RESULTS: Baseline response rate was 70.3%. Depression predictors of diabetes incidence were 'symptomatic' (2.29 [1.28,4.10]), 'symptomatic or ADM use' (2.13 [1.32,3.44]) and 'symptomatic and no ADM use' (2.38 [1.28,4.45]), after adjustment for significant covariates. Being asymptomatic was not a protective factor among those prescribed antidepressants. CONCLUSIONS: Older people with depressive symptoms are at least twice more likely to develop diabetes than those without depressive symptoms, regardless of antidepressants. PMID- 19806605 TI - Expressed emotion in relatives of patients with eating disorders following skills training program. AB - OBJECTIVE: The first aim of this work was to examine whether high expressed emotion (EE) would be associated with psychological distress and negative appraisal of caregiving. The second aim was to examine whether could replicate the findings of (Uehara et al., Compr Psychiatr, 42, 132-138, 2001) regarding a reduction in EE in association with a psycho-educational intervention. METHOD: A prospective exploratory follow-up assessment of 47 caregivers in which EE was measured by Five Minutes Speech Sample, Experience of Caregiving Inventory and General Health Questionnaire (GHQ-12). RESULTS: High EE ratings were found in 26/47 (55%) carers at baseline. The high EE group had a higher negative aspects of caregiving (high EE = 109.8 vs. low EE = 93.7, p = 0.04). The number of carers with high-EE fell from 55% to 24% after intervention, and remained below at follow-up. The levels of distress and caregiving burden of the caregivers were also reduced. DISCUSSION: Fifty-five percent of the caregivers have high EE. High EE relatives noted a higher caregiving burden than low-EE relatives particularly on the 'dimension of loss'. The level of EE was reduced following a skills training program. PMID- 19806606 TI - Dysregulated eating behaviors in borderline personality disorder: are rejection sensitivity and emotion dysregulation linking mechanisms? AB - OBJECTIVE: Individuals with borderline personality disorder (BPD) often engage in dysregulated eating behaviors, such as binge-eating and purging. Rejection sensitivity, or the tendency to worry about and expect rejection in most situations, may be involved in this relationship by increasing the intensity and frequency of emotion dysregulation. METHOD: Using a sample which included individuals diagnosed with BPD, a structural equation model was constructed using BPD symptoms and measures of rejection sensitivity, emotion dysregulation, and dysregulated eating behaviors. RESULTS: The hypothesized model was supported in which BPD symptoms predicted high levels of rejection sensitivity, which then led to increased problems with emotion dysregulation and subsequent dysregulated eating behaviors. A significant indirect effect for rejection sensitivity on dysregulated eating behaviors, through emotion dysregulation, was found. This model also provided better fit than alternative models. DISCUSSION: The results of this study indicate that those with BPD may be more sensitive to rejection, and these fears of rejection may result in increased emotion dysregulation and subsequent dysregulated eating behaviors. Appearance-relevant rejection sensitivity may be an important factor to explore in future research. PMID- 19806607 TI - A qualitative study of perceived social barriers to care for eating disorders: perspectives from ethnically diverse health care consumers. AB - OBJECTIVE: The study aim was to identify and describe health consumer perspectives on social barriers to care for eating disorders in an ethnically diverse sample. METHOD: We conducted an exploratory secondary analysis of qualitative data comprising transcripts from semi-structured interviews with past and prospective consumers of eating disorder treatment (n = 32). Transcripts were inputted into NVivo 8 for coding, sorting, and quantifying thematic content of interest within strata defined by ethnic minority and non-minority participants. We then examined the influence of key social barriers-including stigma and social stereotypes-on perceived impact on care. RESULTS: The majority of respondents (78%) endorsed at least one social barrier to care for an eating or weight concern. Perceived stigma (or shame) and social stereotyping-identified both within social networks and among clinicians-had adversely impacted care for 59% and 19% of respondents, respectively. DISCUSSION: Social barriers to care for eating and weight related concerns may be prevalent in the U.S. and impact both ethnic minority and non-minority health care consumers. PMID- 19806608 TI - Behavioral assessment of the reinforcing effect of exercise in women with anorexia nervosa: further paradigm development and data. AB - OBJECTIVE: To use a Progressive Ratio (PR) computerized "work" paradigm to measure course and correlates of exercise motivation in in-patients with Anorexia Nervosa (AN). METHOD: Sixteen inpatients with AN participated in a PR task assessing the relative reinforcing effect of two different increments of cash versus the opportunity to exercise for upto 30 min, twice; at low weight, and, for n = 10 participants, after weight restoration. RESULTS: There was a trend toward a higher work for exercise with 2 versus 5 increments of cash as the alternative reinforcer. Exercise breakpoint did not differ between low and normal weight states. Exercise breakpoint at each time point was correlated with prehospitalization exercise "commitment" (Commitment to Exercise Scale, r = .613, p = .012 at T1; r = .634, p = .049 at T2). DISCUSSION: Patients with AN will work at a PR task for access to even a small amount of exercise. Exercise motivation during hospitalization is correlated with prehospital exercise commitment and does not appear to change consistently with weight restoration. PMID- 19806609 TI - Cost-utility of treatment of bulimia nervosa. AB - OBJECTIVE: The costs of treating eating disorders are high. Our objective was to perform a cost-utility analysis of treatment of bulimia nervosa. METHOD: 72 patients entering treatment of bulimia nervosa (ICD-10 diagnosis) completed the 15D health-related quality of life (HRQoL) questionnaire and the Eating Disorder Inventory (EDI) before and 6 months after the start of treatment. Quality adjusted life years (QALYs) gained were calculated and cost-utility was assessed within the time horizon of 10 years. RESULTS: Baseline HRQoL was severely impaired in the patients. As a consequence of treatment, mean HRQoL improved clinically and statistically significantly. The cost per QALY gained varied from ?1,455 to ?16,481 (from ?4,428 to ?19,663 discounted at 5%) depending on the assumptions used in the analysis. DISCUSSION: HRQoL of bulimia nervosa patients is severely impaired, but treatment has a clear positive effect on HRQoL. The cost per QALY gained is comparable to many other treatments. PMID- 19806610 TI - Childhood risk factors in Korean women with anorexia nervosa: two sets of case control studies with retrospective comparisons. AB - OBJECTIVE: The aim of this study was to investigate the characteristics of the risk factors for anorexia nervosa (AN) in Korean women. METHOD: Two sets of case control comparisons were conducted, in which 52 women with lifetime AN from Seoul, S. Korea, were compared with 108 Korean healthy controls and also with 42 women with lifetime AN from the UK in terms of their childhood risk factors. A questionnaire designed to conduct a retrospective assessment of the childhood risk factors was administered to all participants. RESULTS: The Korean AN women were more likely to report premorbid anxiety, perfectionism, and emotional undereating and were less likely to report having supportive figures in their childhood than the Korean healthy controls. There were no overall differences in the childhood risk factors between the Korean and British women with AN. DISCUSSION: Premorbid anxiety, perfectionism, less social support, and emotional undereating merit attention as risk factors in Korean AN. The current results are informative, but an epidemiologically robust prospective case-control study would be needed to validate these findings. PMID- 19806611 TI - Virtobot--a multi-functional robotic system for 3D surface scanning and automatic post mortem biopsy. AB - BACKGROUND: The Virtopsy project, a multi-disciplinary project that involves forensic science, diagnostic imaging, computer science, automation technology, telematics and biomechanics, aims to develop new techniques to improve the outcome of forensic investigations. This paper presents a new approach in the field of minimally invasive virtual autopsy for a versatile robotic system that is able to perform three-dimensional (3D) surface scans as well as post mortem image-guided soft tissue biopsies. METHODS: The system consists of an industrial six-axis robot with additional extensions (i.e. a linear axis to increase working space, a tool-changing system and a dedicated safety system), a multi-slice CT scanner with equipment for angiography, a digital photogrammetry and 3D optical surface-scanning system, a 3D tracking system, and a biopsy end effector for automatic needle placement. A wax phantom was developed for biopsy accuracy tests. RESULTS: Surface scanning times were significantly reduced (scanning times cut in half, calibration three times faster). The biopsy module worked with an accuracy of 3.2 mm. DISCUSSION: Using the Virtobot, the surface-scanning procedure could be standardized and accelerated. The biopsy module is accurate enough for use in biopsies in a forensic setting. CONCLUSION: The Virtobot can be utilized for several independent tasks in the field of forensic medicine, and is sufficiently versatile to be adapted to different tasks in the future. PMID- 19806612 TI - PAX5 activates the transcription of the human telomerase reverse transcriptase gene in B cells. AB - Telomerase is an RNA-dependent DNA polymerase that synthesizes telomeric DNA. Its activity is not detectable in most somatic cells but it is reactivated during tumorigenesis. In most cancers, the combination of hTERT hypermethylation and hypomethylation of a short promoter region is permissive for low-level hTERT transcription. Activated and malignant lymphocytes express high telomerase activity, through a mechanism that seems methylation-independent. The aim of this study was to determine which mechanism is involved in the enhanced expression of hTERT in lymphoid cells. Our data confirm that in B cells, some T cell lymphomas and non-neoplastic lymph nodes, the hTERT promoter is unmethylated. Binding sites for the B cell-specific transcription factor PAX5 were identified downstream of the ATG translational start site through EMSA and ChIP experiments. ChIP assays indicated that the transcriptional activation of hTERT by PAX5 does not involve repression of CTCF binding. In a B cell lymphoma cell line, siRNA-induced knockdown of PAX5 expression repressed hTERT transcription. Moreover, ectopic expression of PAX5 in a telomerase-negative normal fibroblast cell line was found to be sufficient to activate hTERT expression. These data show that activation of hTERT in telomerase-positive B cells is due to a methylation-independent mechanism in which PAX5 plays an important role. PMID- 19806614 TI - Oxidation of end-capped pentathienoacenes and characterization of their radical cations. AB - A detailed investigation of the optical and electrochemical properties of two pentathienoacene derivatives, 2,6-bis(trimethylsilyl)-alpha-pentathienoacene (TMS T5-TMS) and 2,6-bis(triisopropylsilyl)-alpha-pentathienoacene (TIPS-T5-TIPS), as the neutral and oxidized species was performed in the temperature range of 80-300 K. The experimental solution UV/Vis and solid-state Raman spectra were interpreted by using time-dependent DFT and DFT quantum chemical calculations at the B3LYP/6-31G** level. Bond lengths, HOMO-LUMO positions, and charge distribution were also predicted by computational methods for both the neutral and oxidized states of each thienoacene. As evidenced by ESR and spectroelectrochemical data, upon oxidation the pentathienoacene derivative with the less sterically hindering trimethylsilyl solubilizing groups, TMS-T5-TMS, undergoes pi dimerization to form [TMS-T5-TMS](2) (2+). In contrast, TIPS-T5 TIPS, with the more bulky triisopropylsilyl solubilizing groups, was oxidized to the radical cation but dimerization was prevented due to steric interactions. These experimental observations are supported by DFT calculations, which were used to investigate [TMS-T5-TMS](2) (2+) and [TIPS-T5-TIPS](2) (2+) pi dimers in the solid state and in solution. The redox potentials and absorption peak locations corresponding to the radical cations and pi dimer of TMS-T5-TMS were identified experimentally. PMID- 19806613 TI - Association study of SNAP25 and schizophrenia in Irish family and case-control samples. AB - SNAP25 occurs on chromosome 20p12.2, which has been linked to schizophrenia in some samples, and recently linked to latent classes of psychotic illness in our sample. SNAP25 is crucial to synaptic functioning, may be involved in axonal growth and dendritic sprouting, and its expression may be decreased in schizophrenia. We genotyped 18 haplotype-tagging SNPs in SNAP25 in a sample of 270 Irish high-density families. Single marker and haplotype analyses were performed in FBAT and PDT. We adjusted for multiple testing by computing q values. Association was followed up in an independent sample of 657 cases and 411 controls. We tested for allelic effects on the clinical phenotype by using the method of sequential addition and 5 factor-derived scores of the OPCRIT. Nine of 18 SNPs had P values <0.05 in either FBAT or PDT for one or more definitions of illness. Several two-marker haplotypes were also associated. Subjects inheriting the risk alleles of the most significantly associated two-marker haplotype were likely to have higher levels of hallucinations and delusions. The most significantly associated marker, rs6039820, was observed to perturb 12 transcription-factor binding sites in in silico analyses. An attempt to replicate association findings in the case-control sample resulted in no SNPs being significantly associated. We observed robust association in both single marker and haplotype-based analyses between SNAP25 and schizophrenia in an Irish family sample. Although we failed to replicate this in an independent sample, this gene should be further tested in other samples. PMID- 19806615 TI - Photocontrolled ring-opening polymerization of strained dicarba[2]ferrocenophanes: a route to well-defined polyferrocenylethylene homopolymers and block copolymers. AB - The ring-opening polymerization (ROP) behavior of a variety of substituted 1,1' ethylenylferrocenes, or dicarba[2]ferrocenophanes, is reported. The electronic absorption spectra and tilted solid-state structures of the monomers rac [Fe(eta(5)-C(5)H(4))(2)(CHiPr)(2)] (7), [Fe(eta(5)-C(5)H(4))(2)(C(H)MeCH(2))] (8), and rac-[Fe(eta(5)-C(5)H(4))(2)(CHPh)(2)] (9) are consistent with the presence of substantial ring strain, which was exploited to synthesize soluble, well-defined polyferrocenylethylenes (PFEs) [Fe(eta(5) C(5)H(4))(2)(C(H)MeCH(2))](n) (12) and [Fe(eta(5)-C(5)H(4))(2)(CHPh)(2)](n) (13) through photocontrolled ROP. Polymer chain lengths could be controlled by the monomer-to-initiator ratio up to about 50 repeat units and, consistent with the "living" nature of the polymerizations, sequential block copolymerization with a sila[1]ferrocenophane led to polyferrocenylethylene-polyferrocenylsilane (PFE-b PFS) block copolymers (14 and 15). PFE polymers 12 and 13 showed two reversible oxidation waves, indicative of appreciable FeFe interactions along the polymer backbone. The diblock copolymers were characterized by NMR spectroscopy, GPC analysis, and cyclic voltammetry. PMID- 19806616 TI - Size selectivity of a copper metal-organic framework and origin of catalytic activity in epoxide alcoholysis. AB - {Cu(bpy)(H(2)O)(2)(BF(4))(2)(bpy)} (Cu-MOF; MOF=metal-organic framework; bpy=4,4' bipyridine), with a 3D-interpenetrated structure and saturated Cu coordination sites in the framework, possesses unexpectedly high activity in the ring-opening reaction of epoxides with MeOH, although the reaction rate drops remarkably with more bulky alcohols. This (apparent) size selection and the single Cu(2+) sites in an identical environment of the crystalline matrix resemble zeolites. The real nature of active sites was investigated by attenuated total reflection infrared (ATR-IR), Raman, EPR, and UV/Vis spectroscopies. Cu-MOF has highly dynamic structural properties that respond to MeOH; its framework dimensions change from 3D to 2D by restructuring to a symmetric coordination of four bpy units to Cu. This interaction is accompanied by the partial dissolution of Cu-MOF as multi-Cu clusters, in which Cu(2+) ions are connected with bpy ligands. Although both molecular and surface catalysis contribute to the high rate of alcoholysis, the soluble oligomeric species (Cu(m)bpy(n)) are far more active. Finally, addition of diethyl ether to the reaction mixture induces the reconstruction of dissolved and solid Cu-MOF to the original framework structure, thereby allowing excellent recyclability of Cu-MOF as an apparent heterogeneous catalyst. In contrast, the original Cu-MOF structure is maintained upon contact with larger alcohols, such as iPrOH and tBuOH, thus leading to poor activity in epoxide ring opening. PMID- 19806617 TI - Chiral encapsulation by directional interactions. AB - The complexation of chiral guests in the cavity of dimeric self-assembled chiral capsule 1(2) was studied by using NMR spectroscopy and X-ray crystallography. Capsule 1(2) has walls composed of amino acid backbones forming numerous directional binding sites that are arranged in a chiral manner. The polar character of the interior dictates the encapsulation preferences towards hydrophilic guests and the ability of the capsule to extract guests from water into an organic phase. Chiral discrimination towards hydroxy acids was evaluated by using association constants and competition experiments, and moderate de values were observed (up to 59 %). Complexes with one or two guest molecules in the cavity were formed. For 1:1 complexes, solvent molecules are coencapsulated; this influences guest dynamics and makes the chiral recognition solvent dependent. Reversal of the preferences can be induced by coencapsulation of a nonchiral solvent in the chiral internal environment. For complexes with two guests, filling of the capsule's internal space can be very effective and packing coefficients of up to 70 % can be reached. The X-ray crystal structure of complex 1(2) superset((S)-6)(2) with well-resolved guest molecules reveals a recognition motif that is based on an extensive system of hydrogen bonds. The optimal arrangement of interactions with the alternating positively and negatively charged groups of the capsule's walls is fulfilled by the guest carboxylic groups acting simultaneously as hydrogen-bond donors and acceptors. An additional guest molecule interacting externally with the capsule reveals a possible entrance mechanism involving a polar gate. In solution, the structural features and dynamic behavior of the D(4)-symmetric homochiral capsule were analyzed by variable-temperature NMR spectroscopy and the results were compared with those for the S(8)-symmetric heterochiral capsule. PMID- 19806618 TI - Spectroscopic and mechanistic studies on oxidation reactions catalyzed by the functional Model SR complex for cytochrome P450: influence of oxidant, substrate, and solvent. AB - Kinetic and mechanistic studies on the formation of an oxoiron(IV) porphyrin cation radical bearing a thiolate group as proximal ligand are reported. The SR complex, a functional enzyme mimic of P450, was oxidized in peroxo-shunt reactions under different experimental conditions with variation of solvent, temperature, and identity and excess of oxidant in the presence of different organic substrates. Through the application of a low-temperature rapid-scan stopped-flow technique, the reactive intermediates in the SR catalytic cycle, such as the initially formed SR acylperoxoiron(III) complex and the SR high valent iron(IV) porphyrin cation radical complex [(SR(*+))Fe(IV)=O], were successfully identified and kinetically characterized. The oxidation of the SR complex under catalytic conditions provided direct spectroscopic information on the reactivity of [(SR(.+))Fe(IV)=O] towards the oxidation of selected organic substrates. Because the catalytically active species is a synthetic oxoiron(IV) porphyrin cation radical bearing a thiolate proximal group, the effect of the strong electron donor ligand on the formation and reactivity/stability of the SR high-valent iron species is addressed and discussed in the light of the reactivity pattern observed in substrate oxygenation reactions catalyzed by native P450 enzyme systems. PMID- 19806619 TI - Gold(I)-catalyzed synthesis of highly substituted 2-cyclopentenones from 5 siloxypent-3-en-1-ynes. PMID- 19806620 TI - Synthesis of multiantennary complex type N-glycans by use of modular building blocks. AB - A modular set of oligosaccharide building blocks was developed for the synthesis of multiantennary N-glycans of the complex type, which are commonly found on glycoproteins. The donor building blocks were laid out for the elongation of a core trisaccharide acceptor (beta-mannosyl chitobiose) conveniently protected with a single benzylidene moiety at the beta-mannoside. Through two consecutive regio- and stereoselective couplings the donors gave N-glycans with three to five antennae in high yields. Due to the consistent protection group pattern of the donors the deprotection of the final products can be performed by using a general reaction sequence. PMID- 19806621 TI - Facile purification of porous metal terephthalates with ultrasonic treatment in the presence of amides. AB - A facile purification method for metal-organic frameworks (MOFs), especially the ones containing insoluble 1,4-benzenedicarboxylic acid (terephthalic acid) in the pore, has been suggested. The purification method consists of the treatment of the MOF with amides such as N,N-dimethylformamide, especially under ultrasound treatment. The purification is completed within 1 h at 70 degrees C as has been confirmed by XRD, nitrogen adsorption, FTIR and TGA measurements. The purification method proved to be simple, one-step, fast and energy-efficient. The MOFs purified by the proposed method show high surface area and micropore volume, confirming the efficiency of the method. The proposed method will lead to a new access to activate (for example, to remove carboxylic acids) MOFs that are unstable above around 100 degrees C. Additionally, the method may be used to transform a non-porous MOF-type material into a porous MOF structure. However, adequate solvents will be necessary for the facile purification of MOFs. PMID- 19806622 TI - Enantio- and diastereoselective iodocyclopropanation of allylic alcohols by using a substituted zinc carbenoid. PMID- 19806623 TI - First heterotrimetallic {3 d-4 d-4 f} single chain magnet, constructed from anisotropic high-spin heterometallic nodes and paramagnetic spacers. PMID- 19806625 TI - Foregoing rigidity to achieve greater intimacy. PMID- 19806626 TI - Artificial protein block copolymers blocks comprising two distinct self assembling domains. PMID- 19806627 TI - Exciton migration in conjugated dendrimers: a joint experimental and theoretical study. AB - We report a joint experimental and theoretical investigation of exciton diffusion in phenyl-cored thiophene dendrimers. Experimental exciton diffusion lengths of the dendrimers vary between 8 and 17 nm, increasing with the size of the dendrimer. A theoretical methodology is developed to estimate exciton diffusion lengths for conjugated small molecules in a simulated amorphous film. The theoretical approach exploits Fermi's Golden Rule to estimate the energy transfer rates for a large ensemble of bimolecular complexes in random relative orientations. Utilization of Poisson's equation in the evaluation of the Coulomb integral leads to very efficient calculation of excitonic couplings between the donor and the acceptor chromophores. Electronic coupling calculations with delocalized transition densities revealed efficient coupling pathways in the bulk of the material, but do not result in strong couplings between the chromophores which are calculated for more localized transition densities. The molecular structures of dendrimers seem to be playing a significant role in the magnitude of electronic coupling between chromophores. Simulated diffusion lengths correlate well with the experimental data. The chemical structure of the chromophore, the shape of the transition densities and the exciton lifetime are found to be the most important factors in determining the size of the exciton diffusion length in amorphous films of conjugated materials. PMID- 19806628 TI - A new perspective on the binding power of an electron. PMID- 19806629 TI - Intrinsic neutral and anionic structures of glutathione. AB - Gas-phase intrinsic structures of intact neutral and anionic glutathione (GSH) have been determined by means of a combination of negative ion photo-electron spectroscopy and quantum chemistry calculations. The inferred structures of the neutral parents of those peptide anions are canonical (non-zwitterionic). These intrinsic structures are compared to those already known in aqueous solution or determined by crystallography in binding sites of enzymes. PMID- 19806630 TI - Carbonates: ecofriendly solvents for palladium-catalyzed direct 2-arylation of oxazole derivatives. PMID- 19806631 TI - Modulation of substrate-membrane interactions by linear poly(2-methyl-2 oxazoline) spacers revealed by X-ray reflectivity and ellipsometry. AB - Hydrated polymer interlayers between planar lipid membranes and solid substrates provide a water reservoir and thus maintain a finite membrane-substrate distance. Linear polymer spacers attached to lipid head groups (lipopolymer tethers) can be used as a defined model of oligo- and polysaccharides covalently anchored on cell surfaces (glycocalyx). They can offer a unique advantage over membranes physisorbed on polymer films (called polymer-cushioned membranes), owing to their ability to control both the length and density of polymer chains. In this study, a lipopolymer tether composed of a stable ether lipid moiety and a hydrophilic poly(2-methyl-2-oxazoline) spacer with a length of 60 monomer units is used to fabricate supported membranes by the successive deposition of proximal (lower) and distal (upper) leaflets. Using specular X-ray reflectivity and ellipsometry, we systematically investigate how the lateral density of polymer chains influences the membrane-substrate interactions. The combination of two types of reflectivity techniques under various conditions enables the calculation of quantitative force-distance relationships. Such artificial membrane systems can be considered as a half-model of cell-cell contacts mediated via the glycocalyx, which reveals the influence of polymer chain density on the interplay of interfacial forces at biological interfaces. PMID- 19806632 TI - Virtual screening against p50 NF-kappaB transcription factor for the identification of inhibitors of the NF-kappaB-DNA interaction and expression of NF-kappaB upregulated genes. AB - Virtual screening against NF-kappaB p50 using docking simulations was applied by starting from a three-dimensional (3D) database containing more than 4.6 million commercially available structures. This database was filtered by specifying a subset of commercially available compounds sharing a (2E,Z)-3-(2-hydroxyphenyl)-2 propenoate substructure and relevant druglike properties. Docking to p50 NF kappaB was performed with a test set of six known inhibitors of NF-kappaB-DNA interactions. In agreement with docking results, the highest-scored compound displayed a high level of inhibitory activity in electrophoretic mobility shift assay (EMSA) experiments (inhibition of NF-kappaB-DNA interactions) and on biological functions dependent on NF-kappaB activity (inhibition of IL-8 gene expression in cystic fibrosis IB3-1 cells). We found that this in silico screening approach is suitable for the identification of low-molecular-weight compounds that inhibit NF-kappaB-DNA interactions and NF-kappaB-dependent functions. Information deduced from the discovery of the new lead compound and its binding mode could result in further lead optimization resulting in more potent NF-kappaB inhibitors. PMID- 19806633 TI - Photochemical key steps in the synthesis of surfactants from furfural-derived intermediates. AB - Furfural is oxidized to 2[5H]-furanone by using hydrogen peroxide or to 5-hydroxy 2[5H]-furanone by using photo-oxygenation. An amine function is introduced by photochemically induced radical addition of tertiairy amines, some of which carry an n-alkyl side chain as hydrophobic moiety. These amines are produced from fatty aldehydes and cyclic secondary amines. The resulting adducts are transformed into amphoteric surfactants possessing an ammonium and a carboxylate function. Amphoteric (pK(N) and isoelectric point) and surfactant properties such as the critical micelle concentration and the adsorption efficiency are determined. PMID- 19806634 TI - Kinetic simulation of a centrifugal bioreactor for high population density hybridoma culture. AB - Demand for increasingly complex post-translationally modified proteins, such as monoclonal antibodies (mAbs), necessitates the use of mammalian hosts for production. The focus of this article is a continuous centrifugal bioreactor (CCBR) capable of increasing volumetric productivity for mAb production through high density hybridoma culture, exceeding 10(8) cells/mL. At these extreme densities, environmental conditions such as substrate and inhibitor concentrations rapidly change dramatically affecting the growth rate. The development of a kinetic model predicting glucose, mAb, lactate, and ammonium concentrations based on dilution rate and cell density is shown in this article. Additionally, it is found that pH affects both growth rate and viability, and a range of 6.9-7.4 is needed to maintain growth rate above 90% of the maximum. Modeling shows that operating an 11.4 mL CCBR inoculated with 2.0 x 10(7) cells/mL at a dilution rate of 1.3 h(-1), results in a predicted growth rate 82% of the maximum value. At the same dilution rate increasing density to 6.0 x 10(7) cells/mL decreases the predicted growth rate to 60% of the maximum; however, by increasing dilution rate to 6.1 h(-1) the growth rate can be increased to 86% of the maximum. Using the kinetic model developed in this research, the concentration of glucose, mAb, lactate, and ammonium are all predicted within 13% of experimental results. This model and an understanding of how RPM impacts cell retention serve as valuable tools for maintaining high density CCBR cultures, ensuring maximum growth associated mAb production rates. PMID- 19806636 TI - Impact of the prehospital ECG on door-to-balloon time in ST elevation myocardial infarction. AB - BACKGROUND: National guidelines have been set to achieve door-to-balloon time (D2B) is less than 90 minutes to improve outcomes for patients with STEMI. The purpose of this study is to see if a more aggressive approach utilizing prehospital ECGs could improve reperfusion times. METHODS: The EMS personnel obtained a 12-lead ECG during initial assessment in the field from patients with chest pain. The ECG was immediately transmitted to the ER physician by cellular link to a computer receiving station. The ER physician reviewed the ECG digital tracing. The cardiac catheterization laboratory (CCL) team was activated from the patients' homes. Patients were transported from the field directly to the CCL by EMS bypassing the emergency department. RESULTS: The mean D2B for patients with initial ECG in hospital in all three hospitals combined was 90.5 minutes, compared to 60.2 minutes in patients with prehospital ECG. (P < 0.0001). When analyzing the mean D2B in regards to times of presentation, we found a significant reduction in mean D2B in patients presenting during working hours (75 minutes) compared with those presenting during off hours (98 minutes) in the control group. However, with the use of prehospital ECGs, there was a significant reduction in D2B regardless of what time the patient arrived in the ER. CONCLUSIONS: Utilizing the prehospital ECG as a tool to bypass ER triage significantly decreases D2B times in patients with STEMI. This technology has the potential to substantially expedite reperfusion therapy in patients with STEMI. PMID- 19806637 TI - Regarding the study In vitro and in vivo studies on thermistor-based intracoronary temperature measurements: effect of pressure and flow. PMID- 19806638 TI - Side-branch wire entrapment during bifurcation PCI: avoidance and management. AB - An LAD/D1 bifurcation intervention was complicated by side-branch wire entrapment and unravelling requiring goose-neck snare removal. Residual microfilaments were retrieved from the main branch after further balloon inflations with a satisfactory final angiographic result and one-year follow-up. Various methods are available to avoid and deal with this complication. PMID- 19806639 TI - A prospective feasibility trial to determine the significance of the sentinel node gradient in breast cancer: a predictor of nodal metastasis location. PMID- 19806640 TI - Long-term surgical outcomes of temporal lobe epilepsy associated with low-grade brain tumors. AB - BACKGROUND: Tumor-related temporal lobe epilepsy (TLE) has a high likelihood of medical intractability and requires surgical treatment. The aims of this study were to analyze the long-term surgical outcomes of and to present appropriate surgical strategies for tumor-related TLE. METHODS: The clinical data of 87 consecutive patients diagnosed with tumor-related TLE were analyzed. The median age at surgery was 22 years. Sixteen patients had a tumor confined to the amygdala or the parahippocampal gyrus, and 10 of them received a tailored lesionectomy without hippocampectomy. The surgical outcome was evaluated based on 3 aspects: seizure control, tumor control, and discontinuation of antiepileptic drugs (AEDs). RESULTS: The actuarial seizure and tumor control rates at the fifth year postoperatively were 79% and 90%, respectively. Seizure control was highly correlated with tumor control. The following factors were found to be significantly associated with poor seizure control: duration of epilepsy>10 years, presence of a remote focus on surface electroencephalography, and incomplete tumor removal. The actuarial AED maintenance rates were 47% at the second year and 11% at the fifth year. The median time to AED discontinuation was 22 months. A younger age at surgery was found to be significantly associated with an increased chance of AED discontinuation. Tailored resection focusing on the tumor resulted in a favorable outcome, even for tumors confined to the amygdala or the parahippocampal gyrus. CONCLUSIONS: Surgical treatment of tumor-related TLE resulted in long-term seizure control in the majority of patients. Maximal tumor removal can be recommended for tumor-related TLE. PMID- 19806641 TI - Leukocytosis at diagnosis and the risk of subsequent thrombosis in patients with low-risk essential thrombocythemia and polycythemia vera. AB - BACKGROUND: Advanced age and a history of thrombosis were well-established risk factors for thrombosis in essential thrombocythemia (ET) and polycythemia vera (PV); cytoreductive therapy was indicated in their presence. Recent studies have suggested leukocytosis as an additional risk factor; however, such an association would be treatment-relevant in the context of low-risk disease. METHODS: In this retrospective study, a Cox proportional hazards model was used to determine the impact of various clinical and laboratory variables, including leukocytosis, on thrombosis-free survival (TFS). Arterial-specific or venous-specific TFS curves for different leukocyte count-defined risk groups were constructed by the Kaplan Meier method and compared using the log-rank test. RESULTS: A total of 407 low risk patients (254 with ET and 153 with PV) were considered. After a respective median follow-up of 104 months and 130 months, respectively, 47 (19%) patients with ET and 41 (27%) with PV experienced a total of 55 (41 arterial and 14 venous) and 46 (22 arterial and 24 venous) thrombotic events, respectively. Leukocytosis at the time of diagnosis, defined by a cutoff level of either 15 or 9.4x10(9)/L, did not appear to be predictive of either arterial or venous thrombosis during follow-up; similar results were obtained when analysis was restricted to patients with platelet counts of <1000x10(9)/L. Instead, advanced age was found to be significantly associated with arterial thrombosis in patients with PV and higher hemoglobin level with venous thrombosis in patients with ET. CONCLUSIONS: In the current retrospective study, leukocytosis at diagnosis did not appear to influence the risk of thrombosis in either ET or PV. However, a prospective study is required before leukocytosis is taken into account during treatment decisions in these disorders. PMID- 19806642 TI - Membranous labyrinth volumes in normal ears and Meniere disease: a three dimensional reconstruction study. AB - OBJECTIVES: The purpose of this study was to determine the normal volume ranges of cochlear duct, saccule, and utricle, and to assess endolymphatic hydrops in Meniere disease. STUDY DESIGN: Retrospective temporal bone study. METHODS: Three dimensional (3-D) images of membranous labyrinth were reconstructed from 31 normal temporal bones, six temporal bones from three patients with bilateral Meniere disease, and 16 temporal bones from eight patients with unilateral Meniere disease. Volumes of each part of membranous labyrinth were measured in each temporal bone group after 3-D reconstruction. RESULTS: The mean volumes and upper normal volume limits (over the 95% confidence interval) of the cochlear duct, saccule, and utricle were 7.67 and 9.77 mm(3), 2.42 and 3.68 mm(3), and 10.65 and 16.45 mm(3), respectively. All three patients with bilateral Meniere disease showed endolymphatic hydrops (excess of volume over normal limits) in both ears. Of eight patients with unilateral Meniere disease, five had no symptom in the contralateral ear, whereas three patients had histories of progression from unilateral to bilateral Meniere disease 13-21 years after the initial onset. All of the diseased and three of eight contralateral ears showed endolymphatic hydrops. In contrast, no hydrops was observed in any part of the membranous labyrinth in asymptomatic ears. CONCLUSIONS: Our findings suggest that cochleosaccular hydrops is a sensitive finding in Meniere disease. In addition, the volume data obtained from this study could be useful as a standard value for the assessment of hydrops in diagnostic imaging of the inner ear in Meniere disease. PMID- 19806643 TI - Clinical utility of postchemoradiation endoscopic brush cytology and biopsy in predicting residual esophageal adenocarcinoma. AB - BACKGROUND: Esophageal adenocarcinoma generally carries a poor prognosis. Treatment with combination chemoradiation (CRT) followed by esophagectomy is becoming common. A pathologic complete response is uncommon but predicts improved survival. Identifying the subset of patients with residual carcinoma has potential management implications. Post-CRT endoscopic brush cytology and biopsy may detect residual tumor; however, the accuracy and clinical value of these methods remain unclear. METHODS: Sixty-seven patients with esophageal adenocarcinoma who underwent preoperative CRT and post-CRT endoscopic brush cytology and biopsy followed by esophagectomy were identified. By using esophagectomy histology as the gold standard, the performance of cytology and biopsy was evaluated in diagnosing residual carcinoma. Two pathologists independently reviewed all false-negative and false-positive cases and resolved disagreements by consensus. RESULTS: The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of cytology for diagnosing residual carcinoma were 26%, 95%, 92%, 35%, and 45%, respectively. For biopsy, these rates were 13%, 90%, 75%, 31%, and 36%, respectively. Sampling error accounted for false-negative diagnoses in approximately 66% of cytology analyses and 98% of biopsy analyses. Approximately 33% of false-negative cytology analyses and 1 false-negative biopsy analysis were caused by the under recognition of tumor cells. Major diagnostic pitfalls included obscuring acute inflammation, necrosis, tumor cells that mimicked benign cells with radiation/reactive atypia, and the under recognition of mucin-containing adenocarcinoma cells. CONCLUSIONS: Brush cytology and biopsy were specific but not sensitive methods for predicting residual cancer after CRT. However, cytology was superior. The current results indicated that brush cytology can be used alone to diagnose residual esophageal carcinoma, and awareness of specific diagnostic pitfalls will help pathologists improve its accuracy. PMID- 19806645 TI - Cancer size, histotype, and cellular grade may limit the success of fine-needle aspiration cytology for screen-detected breast carcinoma. AB - BACKGROUND: Fine-needle aspiration cytology (FNAC) was adopted as the first-line method to assess breast lesions in the Verona Breast Cancer Screening Program. The radiological and pathological factors relating to the success of FNAC in breast cancer series were evaluated. METHODS: Between July 1999 and June 2004, 418 breast cancers were submitted to FNAC in the Verona Breast Cancer Screening Program. The results of FNAC diagnoses were compared with final histology. The FNAC sensitivity rate, underestimation of malignancy rate, and inadequacy rate were correlated with histotype, size, grading, and radiologic imaging. RESULTS: Of the 418 cancers, 95 were in situ, and 323 were invasive. The sensitivity rate was higher in invasive cancers (P < .001), and the underestimation of malignancy rate was greater in in situ cancers (P = .002). Lobular type cancers had a lower sensitivity rate in invasive and in situ cancers. The sensitivity rate was 100% in medullary, mucinous, and papillary cancers, and no case had inadequate sampling. The underestimation of malignancy rate was higher in tubular carcinoma (18.2%); lobular carcinoma showed a higher inadequacy rate (10.4%). The sensitivity rate was lower and the underestimation of malignancy rate was higher in low-grade carcinomas and in lesions <1 cm (P < .001). The performance of FNAC was not significantly influenced by mammographic imaging of lesions. CONCLUSIONS: Low-grade cancer histotype, cancer size <1 cm, and lobular and tubular histotypes limit the possibility of obtaining positive results by FNAC. Operator experience and multidisciplinary consultation may help in overcoming these limitations. Pathologists must be aware of the limits of FNAC; results must be critically evaluated in light of the triple assessment. PMID- 19806646 TI - Sudden bilateral sensorineural hearing loss after intravenous cocaine injection: a case report and review of the literature. AB - Little is known about the effects of intravenous abuse of cocaine, especially on the inner ear. We report on a 26-year-old man who presented to our outpatient department with a sudden severe hearing loss after intravenous injection of cocaine. The audiogram on admission showed symmetric air conduction levels up to 80 dB at 4 kHz. After treatment with intravenous sodium chloride, prednisolone, and pentoxifylline, the audiogram 2 days later showed a bilateral normacusis. A review of the literature on the topic is given and possible reasons for inner ear damages caused by cocaine are discussed. PMID- 19806647 TI - CK19 and CD10 expression in pancreatic neuroendocrine tumors diagnosed by endoscopic ultrasound-guided fine-needle aspiration cytology. AB - BACKGROUND: CK19 and CD10 are useful markers in the differential diagnosis of pancreatic tumors. The authors evaluated CK19 and CD10 expression in pancreatic neuroendocrine tumors (NETs) obtained by endoscopic ultrasound (EUS)-guided fine needle aspiration (FNA). METHODS: Twenty-eight patients diagnosed with pancreatic NETs based on EUS-FNA cytology were studied retrospectively (2004-2007) for immunohistochemical expression of CK19 and CD10. Immunohistochemistry was performed on cell blocks for each case. The pattern of expression for CD10 (cytoplasmic or membranous) and its intensity (0-2) were noted. The staining of the stromal elements for CD10 was recorded as negative. Cytoplasmic staining in tumor cells and percentage distribution (1+ to 4+) for CK19 were regarded as positive. RESULTS: Twenty-three of 28 (82.14%) NETs showed positive cytoplasmic and/or membranous staining for CD10, and 25 of 28 (89.29%) cases were positive for CK19. CONCLUSIONS: The findings demonstrate the high expression of CD10 and CK19 in pancreatic NETs. This indicates that CD10 and CK19 cannot reliably differentiate NETs from other tumors with similar cytomorphologic features (solid pseudopapillary tumors, which frequently stain with CD10, and pancreatic adenocarcinoma, which stains with CK19). PMID- 19806648 TI - Segmental hemangiomas of the upper airway. AB - OBJECTIVES/HYPOTHESIS: To characterize the anatomic distribution of segmental hemangiomas of the larynx and to describe indications for treatment modalities. STUDY DESIGN: Retrospective chart review. METHODS: We performed a retrospective chart review of patients with cutaneous hemangiomas at a tertiary care center over a 4-year period. Only patients with upper airway hemangiomas were studied. We reviewed the anatomic distribution of hemangiomas within the upper airway and the treatment course of each patient. RESULTS: Of 1,226 patients with cutaneous hemangiomas, 108 (9%) were segmental in distribution. There were 56 patients (52%) who had a V3 distribution pattern, and 16 patients (29%) with upper airway involvement. All of these patients had associated V3 or mandibular segmental hemangiomas. As with the cutaneous manifestation, the distribution of hemangioma within the upper airway was segmental. This included the following anatomic sites: oral cavity, oropharynx, hypopharynx, intrinsic structures of the larynx, and subglottis. Many of these patients had diffuse mucosal involvement, including tracheal involvement. A total of 13 out of 16 patients underwent medical intervention, and seven also required surgical intervention. Medical management included systemic (12 patients) and intralesional (two patients) steroids. One patient received chemotherapy prior to referral. Surgical treatment included tracheostomy (four patients prior to referral) and laser ablation of subglottic involvement (total of four patients). CONCLUSIONS: A high percentage of patients with V3 cutaneous hemangiomas (29%) will manifest with upper airway involvement, the distribution of which is segmental. Treatment should take this diffuse pattern of involvement into consideration. PMID- 19806649 TI - A contemporary review of balance dysfunction following vestibular schwannoma surgery. AB - OBJECTIVES/HYPOTHESIS: This review aims to evaluate the literature pertaining to subjective balance dysfunction following vestibular schwannoma surgery; the effect of postoperative imbalance on disability, handicap and quality of life; and to determine factors that influence vestibular compensation. METHODS: Ovid MEDLINE, Cochrane databases, and relevant contemporary texts were searched for papers relating to subjective balance dysfunction following vestibular schwannoma surgery. The quality of this clinical evidence was evaluated. RESULTS: The search yielded 26 studies assessing subjective balance dysfunction following vestibular schwannoma surgery. Analysis revealed that the majority of patients complain of balance dysfunction following surgery; however, a small number report disability or handicap. A few studies have demonstrated a decreased quality of life due to balance dysfunction. Factors have been identified that may contribute to a poor recovery. CONCLUSIONS: Further study is needed of the factors that influence vestibular compensation following vestibular schwannoma surgery. This will help to counsel patients prior to surgery and develop strategies for rehabilitation. PMID- 19806650 TI - Tissue-engineered trachea for airway reconstruction. AB - OBJECTIVES/HYPOTHESIS: Scaffold-free cartilage has been used to engineer biocompatible and mechanically stable neotracheas in vivo. The purpose of this animal study was to determine if neotracheal constructs, implanted paratracheally, could successfully be used for segmental tracheal reconstruction. STUDY DESIGN: Animal study. METHODS: Culture-expanded auricular rabbit chondrocytes were used to engineer scaffold-free cartilage sheets. Cartilage and a strap muscle flap were wrapped around a tube and implanted paratracheally. At 12 to 14 weeks postimplantation neotracheas were used to reconstruct 20 mm tracheal defects. Surgical technique was modified several times in an attempt to decrease the amount of neotracheal obstruction and fibrosis. In one of the six rabbits, neotrachea with its intact strap muscle flap was dropped into the defect followed by an end-to-end anastomosis; in two animals the muscle flap was partially, and in one rabbit completely removed. In two animals the muscle flap was partially removed, the tube reinserted, and the construct reimplanted for 5 weeks to allow formation of a fibrous lining over the exposed cartilage followed by tracheal reconstruction. RESULTS: All implants developed into vascularized and mechanically sound neotracheas. Following reconstruction, none of the animals showed immediate signs of respiratory distress; however, one died after 24 hours due to extensive endotracheal muscle flap edema, whereas rabbits who had undergone partial or complete muscle flap removal survived up to 39 days before developing cicatricial stenosis. CONCLUSIONS: Tissue-engineered neotracheas proved to have excellent biocompatibility and stability to function under physiologic conditions, but lacked adequate endotracheal lining resulting in neotracheal stenosis. PMID- 19806651 TI - The mandibulotomy: friend or foe? Safety outcomes and literature review. AB - OBJECTIVE/HYPOTHESIS: To determine the safety outcomes of a unique mandibulotomy technique and to compare results to the world literature. STUDY DESIGN: Retrospective review of a tertiary care head and neck cancer practice. METHODS: A total of 220 consecutive lip-splitting mandibulotomy access cases from 1998 to 2006 were identified in the University of Alberta's prospective head and neck surgery database and reviewed with follow-up to June 2009. Uniform surgical technique consisting of a lower lip-splitting incision, incisor extraction, a paramedian stair-step osteotomy, and combination fixation with direct interosseous wires and a compression miniplate was utilized for all cases. Variations from traditional methods include adapting the compression miniplate to the reapproximated, rather than precut, mandible and utilizing a mentalis-wire tacking stitch. The main outcome was the complication rate. Complications were recorded and separated into categories consisting of 1) fixation failure: malunion, nonunion, mandibular fracture, plate failure, wire protrusion; and 2) poor wound healing: hardware exposure, orocutaneous fistulae, osteomyelitis, and osteoradionecrosis. RESULTS: Twenty-three (10.5%) mandibulotomy-related complications occurred in 22 (10.0%) patients. Six (2.7%) cases of fixation failure and 17 (7.7%) cases of poor wound healing were identified. The most common complication was hardware exposure. Uni- and multivariate regression analysis failed to show that any patient, tumor, or perioperative variables were statistically significant predictors of complications. Kaplan-Meier analysis showed complications rates of 5.1% at 6 months, 7.0% at 12 months, and 10.2% at 24 months. CONCLUSIONS: The lip-splitting mandibulotomy technique employed provides a safe and effective means of accessing difficult to reach anatomy of the upper aerodigestive tract. PMID- 19806652 TI - A contemporary review of sublingual immunotherapy. AB - The sublingual route has become an interesting and novel therapeutic option for the immunotherapeutic management of patients with allergies. Immunotherapy modifies the immune response by decreasing the specific IgE levels and Th2-type inflammation in the mucosa when allergen exposure occurs, shifting this toward a Th1-type response. Credible evidence exists of both effectiveness and safety of sublingual immunotherapy (SLIT) from several placebo-controlled double-blind studies. SLIT has been shown to be an effective treatment of allergic rhinitis and asthma in both children and adults. The therapy is well tolerated with mainly minor gastrointestinal side effects that subside in few weeks. The ideal treatment length and dosage still require further verification. Additional studies evaluating long-term efficacy and the immune response of SLIT still need to be performed, and additional standardized antigens still need to be developed. PMID- 19806653 TI - Effects of smoking on quality of life following sinus surgery: 4-year follow-up. AB - OBJECTIVES/HYPOTHESIS: The purpose of this study is to report on long-term outcomes in a prospective study comparing the quality-of-life outcomes in smokers and nonsmokers undergoing functional endoscopic sinus surgery. STUDY DESIGN: : Prospective clinical trial. METHODS: A total of 235 patients were prospectively enrolled at a single tertiary academic center. Preoperative Sino-Nasal Outcomes Test (SNOT)-20 scores and comprehensive demographic and smoking data were obtained. Preoperative SNOT-20 scores were compared to postoperative SNOT-20 scores. RESULTS: Long-term follow-up results were available in 111/185 nonsmokers (60%) and 26/50 smokers (52%). Nonsmokers had a mean preoperative SNOT-20 score of 26.2, a short-term follow-up score of 10.1 (mean, 3.1 months) and a long-term follow-up score of 11.4 (mean, 44.2 months). Smokers had a mean preoperative SNOT 20 score of 27.8, a short-term follow-up score of 5.7 (mean, 3.1 months) and a long-term follow-up score of 8.3 (mean, 40.3 months). The mean improvement in SNOT-20 scores for smokers was significantly greater than nonsmokers in short term follow-up (P = .044), however, no significant difference in improvement was seen between smokers and nonsmokers at long-term follow-up. CONCLUSIONS: Both smokers and nonsmokers continue to maintain a highly significant improvement in SNOT-20 scores following endoscopic sinus surgery at long-term follow-up. Although smoking remains a well-documented cause of medical morbidity, smokers maintained an improvement in quality of life after long-term follow-up from endoscopic sinus surgery in this prospective study. PMID- 19806654 TI - Solitary fibrous tumor of the thyroid gland. AB - We report a case of solitary fibrous tumor of the thyroid gland in a 51-year-old man with pre-existing right recurrent laryngeal nerve paralysis from a congenital tracheoesophageal fistula repair as an infant. The left thyroid lobe was enlarged and soft. Fine needle aspiration biopsy via ultrasound guidance demonstrated a hypercellular aspirate composed of spindle cells with bland nuclear morphology. Given this histomorphology and immunohistochemical profile, the diagnosis of solitary fibrous tumor was considered and confirmed following left thyroid lobectomy. PMID- 19806655 TI - Morphology of the quadrate in the Eocene anseriform Presbyornis and extant galloanserine birds. AB - Despite the notoriety, phylogenetic significance, and large number of available specimens of Presbyornis, its cranial anatomy has never been studied in detail, and its quadrate has been partly misinterpreted. We studied five quadrates of Presbyornis that reveal features hitherto unknown in the anseriforms but otherwise present in galliforms. As a result, we analyzed the variable quadrate characters among all extant galloanserine families and identified synapomorphies and other morphological variation among the major galloanserine clades. In terms of quadrate morphology, Presbyornis is more plesiomorphic than any extant anseriform (including the Anhimidae) and shares ancestral galloanserine characters with the Megapodiidae, the earliest branch of extant galliforms. The quadrate's morphology is inconsistent with the currently accepted anseriform phylogeny that nests Presbyornis within the crown-group as a close relative of the Anatidae. The presbyornithid quadrates exhibit an unusual variation in the presence of a caudomedial pneumatic foramen, which we interpret as a result of a discontinuous change in the growth path of the pneumatic diverticulum. Another episode of morphogenetic imbalance in the growth path of the pneumatic diverticulum may have accompanied the disappearance of the basiorbital pneumatic foramen (along with the pneumatization of the pterygoid) at the origin of the crown-group anseriforms. This episode is marked by the striking individual variation in the presence and location of pneumatic foramina in the mandibular part of the quadrate in the Anhimidae. PMID- 19806656 TI - The constrictor dorsalis musculature and basipterygoid articulation in Sphenodon. AB - The constrictor internus dorsalis (CID) trigeminal muscles in vertebrates lie between the braincase and the palatoquadrate bar, and in reptiles they are believed to function in the relative movements between braincase and maxillary segment known as kinesis. In amniote taxa, the presence of a synovial basipterygoid articulation (BPA) correlates with presence of the CID. Previous descriptions of the CID in the tuatara Sphenodon (Rhynchocephalia) are inconsistent regarding presence, size and direction, particularly of the m. protractor pterygoidei. The Sphenodon skull is reported to be akinetic. The CID was investigated in Sphenodon by examination of sectioned embryonic and hatchling material, and by dissection of fixed specimens. Osteological correlates of CID muscles and morphology of the BPA were examined on disarticulated skulls and on CT scan images. The vectors of action of these muscles in relation to the BPA were projected onto CT images. Mm. levator bulbi and levator pterygoidei are found to be similar to most previous descriptions, but m. protractor pterygoidei was found in a different position, lying entirely medial to the palatoquadrate bar. The insertions of mm. levator pterygoidei and protractor pterygoidei are visible on the disarticulated pterygoid bone. The BPA is mobile only by rotation around the horizontal axis of the joints themselves; metakinesis is not possible in the Sphenodon skull. M. protractor pterygoidei appears to either resist or recognize lateral displacement of the BPA. M. levator ptergyoidei is placed to resist dorsal displacement of the braincase at the BPA, or torsion of the braincase around its longitudinal axis. The BPA appears to be a means to direct compressive stress via the base of the braincase and occipital condyle to the cervical spine in Sphenodon, and probably in its direct ancestors. Metakinesis may never have been a feature of the lepidosaur skull. PMID- 19806657 TI - PKMzeta maintains 1-day- and 6-day-old long-term object location but not object identity memory in dorsal hippocampus. AB - Continuous activity of the atypical protein kinase C isoform M zeta (PKMzeta) is necessary for maintaining long-term memory acquired in aversively or appetitively motivated associative learning tasks, such as active avoidance, aversive taste conditioning, auditory and contextual fear conditioning, radial arm maze, and watermaze. Whether unreinforced, nonassociative memory will also require PKMzeta for long-term maintenance is not known. Using recognition memory for object location and object identity, we found that inactivating PKMzeta in dorsal hippocampus abolishes 1-day and 6-day-old long-term recognition memory for object location, while recognition memory for object identity was not affected by this treatment. Memory for object location persisted for no more than 35 days after training. These results suggest that the dorsal hippocampus mediates long-term memory for where, but not what things have been encountered, and that PKMzeta maintains this type of spatial knowledge as long as the memory exists. PMID- 19806659 TI - The entorhinal cortex, but not the dorsal hippocampus, is necessary for single cue latent learning. AB - Two experiments were conducted to examine the roles of the entorhinal cortex (EC), dorsal hippocampus (DH), and ventral hippocampus (VH) in a modified Latent Cue Preference (LCP) task. The modified LCP task utilized one visual cue in each compartment, compared to several multimodal cues used in a previous version. In the single-cue LCP task, water-replete rats drink water in one compartment of the LCP box on 1 day, and then have no water in a second compartment of the LCP box the following day (one training trial), for a total of three training trials. Rats are then water-deprived prior to a preference test, in which they are allowed to move freely between the two compartments with the water removed. Latent learning is demonstrated when water-deprived rats spend more time in the compartment that previously contained the water. Experiment 1 demonstrated that the single-cue LCP task results in the same irrelevant-incentive latent learning as the multicue LCP task. In addition, Experiment 1 replicated the finding that a compartment preference based on this latent learning requires a deprivation state during the preference test, while a compartment preference based on conditioning does not. Experiment 2 examined the effects of pretraining neurotoxin lesions of the EC, DH, and VH on this single-cue LCP task. Results showed that lesions of the EC and VH disrupted the irrelevant-incentive latent learning, while lesions of the DH did not. These results indicate that a latent learning task that involves one discrete compartment cue, rather than several compartmental cues, does not require the DH. Therefore, the EC appears to play a central role in single-cue latent learning in the LCP task. PMID- 19806658 TI - Hippocampal NMDA receptor subunits differentially regulate fear memory formation and neuronal signal propagation. AB - Activation of NMDA receptors (NMDAR) in the hippocampus is essential for the formation of contextual and trace memory. However, the role of individual NMDAR subunits in the molecular mechanisms contributing to these memory processes is not known. Here we demonstrate, using intrahippocampal injection of subunit selective compounds, that the NR2A-preferring antagonist impaired contextual and trace fear conditioning as well as learning-induced increase of the nuclear protein c-Fos. The NR2B-specific antagonist, on the other hand, selectively blocked trace fear conditioning without affecting c-Fos levels. Studies with cultured primary hippocampal neurons, further showed that synaptic and extrasynaptic NR2A and NR2B differentially regulate the extracellular signal regulated kinase 1 and 2/mitogen- and stress-activated protein kinase 1 (ERK1/2/MSK1)/c-Fos pathway. Activation of the synaptic population of NMDAR induced cytosolic, cytoskeletal, and perinuclear phosphorylation of ERK1/2 (pERK1/2). The nuclear propagation of pERK1/2 signals, revealed by upregulation of the downstream nuclear targets pMSK1 and c-Fos, was blocked by a preferential NR2A but not by a specific NR2B antagonist. Conversely, activation of total (synaptic and extrasynaptic) NMDAR engaged receptors with NR2B subunits, and resulted in membrane retention of pERK1/2 without inducing pMSK1 and c-Fos. Stimulation of extrasynaptic NMDAR alone was consistently ineffective at activating ERK signaling. The discrete contribution of synaptic and total NR2A- and NR2B-containing NMDAR to nuclear transmission vs. membrane retention of ERK signaling may underlie their specific roles in the formation of contextual and trace fear memory. PMID- 19806660 TI - Enhancement of ethanol and biogas production from high-crystalline cellulose by different modes of NMO pretreatment. AB - Pretreatment of high-crystalline cellulose with N-methyl-morpholine-N-oxide (NMO or NMMO) to improve bioethanol and biogas production was investigated. The pretreatments were performed at 90 and 120 degrees C for 0.5-15 h in three different modes, including dissolution (85% NMO), ballooning (79% NMO), and swelling (73% NMO). The pretreated materials were then enzymatically hydrolyzed and fermented to ethanol or anaerobically digested to biogas (methane). The pretreatment at 85% NMO, 120 degrees C and 2.5 h resulted in 100% yield in the subsequent enzymatic hydrolysis and around 150% improvement in the yield of ethanol compared to the untreated and water-treated material. However, the best results of biogas production were obtained when the cellulose was treated with swelling and ballooning mode, which gave almost complete digestion in 15 days. Thus, the pretreatment resulted in 460 g ethanol or 415 L methane from each kg of cellulose. Analysis of the structure of treated and untreated celluloses showed that the dissolution mode can efficiently convert the crystalline cellulose I to cellulose II. However, it decreases the water swelling capacity of the cellulose. On the other hand, swelling and ballooning modes in NMO treatment were less efficient in both water swelling capacity and cellulose crystallinity. No cellulose loss, ambient pressure, relatively moderate conditions, and high efficiency make the NMO a good alternative for pretreatment of high-crystalline cellulosic materials. PMID- 19806661 TI - Secondary clonal cytogenetic abnormalities following successful treatment of acute promyelocytic leukemia. AB - To identify patients who developed secondary clonal cytogenetic aberrations (CCA) following therapy for acute promyelocytic leukemia (APL), we retrospectively analyzed cytogenetic results from 123 patients diagnosed with APL between 1995 and 2007, who had ongoing cytogenetic analysis undertaken in our laboratory. During follow-up for APL we identified 12 patients (9.8%) who developed CCA, not detected at diagnosis of APL and unrelated to their original APL karyotype. All patients had received all-trans retinoic acid (ATRA) and chemotherapy and were in complete remission for APL when secondary CCA were identified. The median latency period between diagnosis of APL and emergence of secondary CCA was 27.5 months (range: 2-54 months). To date, four patients with CCA have been diagnosed with therapy-related myelodysplastic syndrome (t-MDS)/acute myeloid leukemia (t-AML), giving a median t-MDS/AML free survival of 78 months, with follow-up ranging between 20 and 136 months from APL diagnosis. Three patients have died: two patients died of t-AML and another developed relapsed APL with persistence of his secondary clone but no diagnosis of t-MDS/AML and died from transplant-related complications. Two patients are alive with t-MDS. Seven patients with CCA are alive with no morphological evidence of MDS at the time of their last known follow-up; thus median survival has not been reached. The appearance of these abnormalities in the absence of morphological evidence of MDS in the majority of patients is unusual, and highlights the importance of continued cytogenetic follow-up in these patients. Am. J. Hematol., 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19806662 TI - Acute monoblastic leukemia with abnormal granules and disseminated intravascular coagulation: diagnostic pitfalls. PMID- 19806663 TI - Venous thrombosis associated with gene deletion of tissue factor pathway inhibitor. PMID- 19806664 TI - A pediatric case series of acute hemolysis after administration of intravenous immunoglobulin. PMID- 19806665 TI - Salvage therapy for acute myeloid leukemia with fludarabine, cytarabine, and idarubicin with or without gemtuzumab ozogamicin and with concurrent or sequential G-CSF. AB - The current salvage therapies for relapsed/refractory acute myeloid leukemia (AML) are unsatisfactory. Over the past 7 years, we have used two salvage regimens: fludarabine, cytarabine, and idarubicin with (FLAG-IM) or without gemtuzumab ozogamicin (GO) (9 mg/m(2) on Day 8) (FLAG-I) in relapsed/refractory AML. Three-quarters of patients also received concurrent G-CSF. Seventy-one patients were treated, 23 with FLAG-I and 48 with FLAG-IM. The median duration of follow-up was 30.6 months. The treatment groups were well balanced with median ages of 48 years (range 18-70) and 47 years (range 20-68), unfavorable cytogenetics in 57% and 35%, prior allogeneic stem cell transplant in 43% and 42%, and CR1 duration <1 year in 60% and 67%, respectively, for FLAG-I and FLAG IM. The complete remission (CR) rate in the FLAG-I group was 39% with an additional 13% achieving a CRp [overall response rate (ORR) 52%]; the CR rate in the FLAG-IM group was 29% with an additional 27% achieving a CRp (ORR 56%). The median duration of response (DOR; 16.8 vs. 8.3 months), event-free survival (EFS; 7.4 vs. 4.1 months), and overall survival (OS; 8.8 vs. 5.0 months) trended to favor FLAG-I over FLAG-IM. The patients who received G-CSF concurrent with chemotherapy had superior overall response rate (ORR; 62% vs. 29%, P = 0.026), median EFS (6.2 vs. 3.4 months, P = 0.010), and OS (8.8 vs. 3.9 months, P = 0.004) when compared with those who sequentially received G-CSF and chemotherapy, regardless of chemotherapy regimen. The addition of GO, at this dose and schedule, to FLAG-I failed to improve the outcomes in patients with relapsed/refractory AML. The patients who received G-CSF concurrently with chemotherapy had improved outcomes. Am. J. Hematol., 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19806667 TI - CAZIP, a novel protein expressed in the developing heart and nervous system. AB - Recently, we have performed a whole genome micro-array analysis on human embryonic stem cells differentiating toward cardiomyocytes, which resulted in the identification of novel genes that were highly up-regulated during differentiation. Here, we describe one of these novel genes annotated as KIAA0774. The predicted protein contains a leucine-zipper domain at the C terminus and has at least two isoforms (358 and 1354 amino acids). Whole-mount in situ hybridization confirmed that the mRNA of both the mouse and chicken orthologs of KIAA0774 is expressed during early cardiac development. Hence, we named this protein CAZIP (cardiac zipper protein). Later during embryonic development, Cazip was also expressed in parts of the nervous system. Northern blot and real-time polymerase chain reaction analysis showed that Cazip is expressed in heart and brain in adult mice. These results suggest a role for CAZIP in development and function of the heart and nervous system in vertebrates. PMID- 19806666 TI - Different timings of Dicer deletion affect neurogenesis and gliogenesis in the developing mouse central nervous system. AB - MicroRNAs, processed by the RNAase III enzyme Dicer, are approximately 22 nucleotide endogenous noncoding small RNAs. The function of Dicer in the mouse central nervous system (CNS) development is not well understood. Here, we show that specifically deleting Dicer expression in the CNS and in the cerebral cortex using two Cre lines results in reduced progenitor numbers, abnormal neuronal differentiation, and thinner cortical wall. Incomplete Dicer deletion during early embryonic stages contributes to normal development of early-born neurons in the cortex and motor neurons in the spinal cord. However, at late embryonic stages when Dicer is completely ablated in the CNS, the migration of late-born neurons in the cortex and oligodendrocyte precursor expansion and differentiation in the spinal cord are greatly affected. Our studies of different timings of Dicer deletion demonstrate the importance of the Dicer-mediated microRNA pathway in regulating distinct phases of neurogenesis and gliogenesis during the CNS development. PMID- 19806668 TI - Patterns of Wnt pathway activity in the mouse incisor indicate absence of Wnt/beta-catenin signaling in the epithelial stem cells. AB - The Wnt pathway is crucial for tooth development as shown by dental defects caused by impaired Wnt signaling in mouse and human. We investigated Wnt signaling in continuously growing mouse incisors focusing on epithelial stem cells. Ten Wnt ligands were expressed both in the dental epithelium and mesenchyme, and were associated mainly with odontoblast and ameloblast differentiation. Wnt/beta-catenin activity was detected in mesenchyme in BATgal and TOPgal reporter mice while Axin2, also a reporter of Wnt/beta-catenin signaling, was expressed additionally in the epithelium. Axin2 was, however, excluded from the epithelial stem cells in the cervical loop. Interestingly, these cells expressed specifically Lgr5, a Wnt target gene and stem cell marker in the intestine, suggesting that Lgr5 is a marker of incisor stem cells but is not regulated by Wnt signaling in the incisor. We conclude that epithelial stem cells in the mouse incisors are not regulated directly by Wnt/beta-catenin signaling. PMID- 19806669 TI - Anatomy of the floor of the third ventricle in relation to endoscopic ventriculostomy. AB - Neuroanatomical structures that form the floor of the third ventricle (FTV) and neighboring tissues are important in the context of third ventriculostomy (TV) procedures. Thorough knowledge of the anatomical and histological organization of the region would be useful in understanding and preventing surgical complications. Taking the third ventricle region as a model, we aimed to simulate TV and make measurements of 23 cadaver brains, as well as perform histological examinations of the third ventricular floor on five cadaver brains. During the endoscopic TV, we examined the degree to which the structures surrounding the FTV were affected by surgical simulation. To make a clinical comparison, the distance between the center of the FTV and the basilar apex was measured on cranial magnetic resonance images (MRIs) of 15 subjects with normal ventricular systems and 15 subjects with moderately enlarged ventricles. Histological examination revealed that the ependymal cells and arachnoid membrane formed the inner and outer surfaces of the third ventricle floor, respectively, whereas the stroma was made up of glial cells exclusively. This region was gliotic and avascular. When cadaver brains with normal and hydrocephalic ventricles were compared, there were significant differences in the distance between the center of the floor and the basilar apex (P < 0.001). On the basis of our study, the optimal site for TV fenestration and balloon inflation is just anterior to the mamillary bodies to avoid injury to neighboring structures. PMID- 19806670 TI - Anatomical features of the opening of the nasolacrimal duct and the lacrimal fold (Hasner's valve) for intranasal surgery: a cadaveric study. AB - The location and size of the opening of the nasolacrimal duct and the lacrimal fold (Hasner's valve) are variable. These features have clinical importance in nasal surgery because of the difficulty in determining their location. The aim of this study was to demonstrate the anatomical features of the opening of the nasolacrimal duct and the lacrimal fold and to discuss the importance of such knowledge in minimizing the risk of injury at surgery. Twenty sagittal head sections from formalin-fixed cadavers were examined. The sections showed no evidence of pathology or trauma. The type of opening of the nasolacrimal duct was a vertical sulcus in 14 of 20 (70%), an oblique sulcus in 2 of 20 (10%), an oblique fissure in 2 of 20 (10%), a vertical fissure in 1 of 20 (5%), and an anteroposterior fissure in 1 of 20 specimens (5%). The lacrimal fold was present in 16 of 20 specimens (80%). Five different forms of this fold were observed. Some morphological features were evaluated quantitatively. We believe that detailed anatomical knowledge of the opening of nasolacrimal duct will be useful in surgical approaches to this area. PMID- 19806671 TI - An anatomical study of the muscles that attach to the articular disc of the temporomandibular joint. AB - The masticatory muscles are generally described as the muscles that originate from the cranium and insert on the mandible. Some of the masticatory muscles also insert into the articular disc of the temporomandibular joint. Although there are numerous reports of studies on the attachment of the fibers to the disc, most reports discuss only one muscle. We have shown that the masticatory muscles are not simply a group of clearly independent muscles, but that these muscles contain various transitional muscle bundles among the major muscles. From this point of view, we carried out minute dissection of the collective muscles and muscle bundles surrounding the temporomandibular joint. We dissected 40 head halves of 20 Japanese cadavers (10 males, 10 females: average 79.6 yr). After complete removal of the bony elements, the structures surrounding the temporomandibular joint were investigated en-block. In all specimens, the superior surface of the upper head of lateral pterygoid and the midmedial muscle bundle were attached to the disc. In some specimens, the discotemporal bundle, zygomaticomandibularis, and masseter were attached to the anterior surface of the disc. The total vector of these muscles pulls the disc anteriorly. In contrast, the vector of the muscles to the condylar processes of the mandible pulls the mandible medially. From these observations, it seems that the fibers, which attach to the disc act to steady the disc against the masticatory movement. PMID- 19806672 TI - A case of double Gantzer's muscle and its possible role in nerve entrapment. PMID- 19806673 TI - Unusual combination of musculotendinous variations in the plantar musculature. PMID- 19806674 TI - Editorial. PMID- 19806675 TI - Global metabolic profiling of plant cell wall polysaccharide degradation by Saccharophagus degradans. AB - Plant cell wall polysaccharides can be used as the main feedstock for the production of biofuels. Saccharophagus degradans 2-40 is considered to be a potent system for the production of sugars from plant biomass due to its high capability to degrade many complex polysaccharides. To understand the degradation metabolism of plant cell wall polysaccharides by S. degradans, the cell growth, enzyme activity profiles, and the metabolite profiles were analyzed by gas chromatography-time of flight mass spectrometry using different carbon sources including cellulose, xylan, glucose, and xylose. The specific activity of cellulase was only found to be significantly higher when cellulose was used as the sole carbon source, but the xylanase activity increased when xylan, xylose, or cellulose was used as the carbon source. In addition, principal component analysis of 98 identified metabolites in S. degradans revealed four distinct groups that differed based on the carbon source used. Furthermore, metabolite profiling showed that the use of cellulose or xylan as polysaccharides led to increased abundances of fatty acids, nucleotides and glucuronic acid compared to the use of glucose or xylose. Finally, intermediates in the pentose phosphate pathway seemed to be up-regulated on xylose or xylan when compared to those on glucose or cellulose. Such metabolic responses of S. degradans under plant cell wall polysaccharides imply that its metabolic system is transformed to more efficiently degrade polysaccharides and conserve energy. This study demonstrates that the gas chromatography-time of flight mass spectrometry-based global metabolomics are useful for understanding microbial metabolism and evaluating its fermentation characteristics. PMID- 19806676 TI - Theoretical investigation of the sequential reductive dechlorination pathways of chlorobenzenes and chloroanilines. AB - The distribution of product isomers during the sequential reductive dechlorination of pentachloroaniline (PCA) and pentachlorobenzene (PeCB) was examined based on calculated thermodynamic, chromatographic, and electronic properties and then compared to the product distribution achieved by enrichment cultures. The dechlorination pathway analysis based on free energy considerations matched 78% and 67% of the experimental results for the sequential reductive dechlorination of chlorobenzenes (CBs) and chloroanilines (CAs), respectively. Chromatographic properties of CBs and CAs were able to explain some but not all of the reactions in the observed dechlorination pathways. Correlations between the observed dechlorination pattern and electronic properties of the parent compounds were able to explain most of the formation of the observed products. Experimentally observed sequential reductive dechlorination of CBs and CAs were similar to predicted dechlorination pathways based on the charge differential values calculated for the carbon-chloride bonds. Chlorine atoms were removed from the carbon atom that has the highest charge differential or the second highest charge differential. However, although thermodynamic, electronic as well as chromatographic properties of the CBs and CAs are certainly important factors, they may not be sufficient to completely describe the sequential microbial reductive dechlorination. Enzymatic specificity, as well as other factors (i.e., culture acclimation, environmental factors) should be considered for the interpretation of observed sequential reductive dehalogenation pathways of haloorganic compounds. This work provides the most comprehensive analysis to date of theoretical factors that control the sequential reductive chlorination of two homologous series of single-ring chloroaromatic species. PMID- 19806677 TI - Investigating the role of native propionyl-CoA and methylmalonyl-CoA metabolism on heterologous polyketide production in Escherichia coli. AB - 6-Deoxyerythronolide B (6dEB) is the macrocyclic aglycone precursor of the antibiotic natural product erythromycin. Heterologous production of 6dEB in Escherichia coli was accomplished, in part, by designed over-expression of a native prpE gene (encoding a propionyl-CoA synthetase) and heterologous pcc genes (encoding a propionyl-CoA carboxylase) to supply the needed propionyl-CoA and (2S)-methylmalonyl-CoA biosynthetic substrates. Separate E. coli metabolism includes three enzymes, Sbm (a methylmalonyl-CoA mutase), YgfG (a methylmalonyl CoA decarboxylase), and YgfH (a propionyl-CoA:succinate CoA transferase), also involved in propionyl-CoA and methylmalonyl-CoA metabolism. In this study, the sbm, ygfG, and ygfH genes were individually deleted and over-expressed to investigate their effect on heterologous 6dEB production. Our results indicate that the deletion and over-expression of sbm did not influence 6dEB production; ygfG over-expression reduced 6dEB production by fourfold while ygfH deletion increased 6dEB titers from 65 to 129 mg/L in shake flask experiments. It was also found that native E. coli metabolism could support 6dEB biosynthesis in the absence of exogenous propionate and the substrate provision pcc genes. Lastly, the effect of the ygfH deletion was tested in batch bioreactor cultures in which 6dEB titers improved from 206 to 527 mg/L. PMID- 19806678 TI - Isomerization in the CDR2 of a monoclonal antibody: Binding analysis and factors that influence the isomerization rate. AB - Isomerization of a monoclonal antibody is one of the common routes of protein degradation. An isomerization in the complementarity-determining region (CDR) was found previously and is investigated in depth in this work. Affinity analysis proves that the antibody with one isomerized heavy chain has lower binding. Binding constants were determined, and exhibited a slower on-rate in conjunction with a faster off-rate for this isomerization. To determine the role of the buffer on the rate of isomerization, this antibody was incubated in various matrices and the amount of isomerized antibody was determined by hydrophobic interaction chromatography (HIC). The rate was found to be dependent on the pH as well as the net negative charge of the buffer components that can act as proton acceptors. An Arrhenius plot was performed to predict the levels of isomerization and a comparison of real samples proved the model was correct. This work affirms that isomerization in the CDR of a therapeutic antibody is important to monitor and the formulation buffer plays a significant role in the rate of the isomerization. PMID- 19806679 TI - Abstracts of the annual meeting of the American Academy of Otolaryngology--Head and Neck Surgery Foundation, October 4-7, 2009. Philadelphia, Pennsylvania, USA. PMID- 19806680 TI - [Effect of donor liver extract on liver regeneratrion: the experimental results]. PMID- 19806681 TI - [Diaphragm ruptures]. PMID- 19806682 TI - [Gastrointestinal stromal tumors]. PMID- 19806683 TI - [Profuse gastric bleeding in HIV-infected patient]. PMID- 19806684 TI - [Plenary session of problem committees emergency surgery and surgical infection of interdepartmental scientific surgical board of Russian Academy of Medical Sciences and Ministry of Health and Social Development]. PMID- 19806685 TI - Defatting the fatty liver with normothermic perfusion of the liver allograft. PMID- 19806686 TI - Is survival after liver transplantation for hepatocellular carcinoma influenced by the etiology of the liver disease? PMID- 19806687 TI - Should intraportal insulin administration become routine in clinical transplant care? PMID- 19806688 TI - What did you learn at the CAEP conference in Calgary? PMID- 19806689 TI - Nuclear binding of exogenous histones by L cells at low pH. PMID- 19806690 TI - Localization of acid phosphatase in lipofuscin granules and possible autophagic vacuoles in interstitial cells of the guinea pig testis. PMID- 19806691 TI - The effects of osmotic lysis on the oxidative phosphorylation and compartmentation of rat liver mitochondria. PMID- 19806692 TI - Isolation of ribosomes from cysts of Entamoeba invadens. PMID- 19806693 TI - Proteins in nucleocytoplasmic interactions. II. Turnover and changes in nuclear protein distribution with time and growth. PMID- 19806694 TI - III. Electron microscopic radioautographic study of the rat heart perfused with tritiated oleic acid. PMID- 19806695 TI - Some properties of the protein forming the outer fibers of cilia. PMID- 19806696 TI - The effect of puromycin on intranuclear steps in ribosome biosynthesis. PMID- 19806697 TI - The mechanism of mitochondrial extrusion from phenylhydrazine-induced reticulocytes in the circulating blood. PMID- 19806698 TI - The effect of mitotic inhibitors in myogenesis in vitro. PMID- 19806699 TI - Ultrastructural studies on the lymphatic anchoring filaments. PMID- 19806700 TI - The fine structure of the lateral vestibular nucleus in the rat. I. Neurons and neuroglial cells. PMID- 19806701 TI - Effects of actinomycin D and puromycin on the ACTH-induced ultrastructural transformation of mitochondria of cortical cells of rat adrenals in tissue culture. PMID- 19806702 TI - The penetration of reovirus RNA and initiation of its genetic function in L strain fibroblasts. PMID- 19806703 TI - The use of millipore filters in ultrastructural studies of cell cultures and viruses. PMID- 19806705 TI - Ductal excretion of neutral red lysosomes in the mouse pancreas. PMID- 19806704 TI - Obliquely striated muscle. IV. Sarcoplasmic reticulum, contractile apparatus, and endomysium of the body muscle of a polychaete, Glycera, in relation to its speed. PMID- 19806706 TI - Ultrastructure of chloroplasts isolated by nonaqueous extraction. PMID- 19806707 TI - Intercellular bridges and synchronization of germ cell differentiation during oogenesis in the rabbit. PMID- 19806708 TI - The thymidine pool in grasshopper neuroblasts during mitosis. PMID- 19806709 TI - Hospital length of stay: social work services as an important factor. AB - In earlier studies it was found that the severity of patients' psychosocial problems was a significant predictor of length of stay (LOS). This current study compared predictors of LOS for samples of patients referred to social services in three large urban hospitals in June-October 2002 (n = 176) and 2006 (n = 147), and examined changes in patient characteristics and the nature of social work practice. A significant relationship between psychosocial severity and LOS was again found, confirming the important role that social services can potentially play in controlling hospital costs. Some significant changes were also found in the pattern of social work practice; this was generally in the direction of more community consultation and collaboration, suggesting a greater emphasis on multidisciplinary teamwork. PMID- 19806710 TI - The use of public policy analysis to enhance the Nursing Home Reform Act of 1987. AB - Given the prevalence of abuse and neglect in nursing home care delivery vis-a-vis elderly and frail residents, and despite the advent and implementation of the Nursing Home Reform Act of 1987 (a policy that sought to diminish such conduct in such institutional settings), deficiency citations in nursing home care and services remain both problematic and common. By employing public policy analysis, and by analyzing various social science theories applicable to the improvement of care delivery and quality, this article seeks to develop methods to enhance compliance with the Nursing Home Reform Act and reduce care deficiencies in nursing homes. PMID- 19806711 TI - Family predictors of psychosocial outcomes among Hong Kong Chinese cancer patients in palliative care: living and dying with the "support paradox". AB - A good death is universally desired. For Chinese patients, the family is believed to play a key role in making this possible. This study aims at exploring the relationship of family-related factors and psychosocial outcomes among Hong Kong Chinese cancer patients in palliative care. Clinical data mining was adopted as the research method. Nurses collected data from clinical interviews with incoming palliative care patients. A total of 935 patients from three years of deceased patient records was included. Stepwise regression analysis demonstrated that "caregivers" support and acceptance" predicted fewer psychosocial symptoms of patients, whereas "depressed family response to patient's illness" and "family anxiety" predicted a greater number of psychosocial symptoms of patients upon their admission to palliative care. The findings suggested two possible pathways toward enhancing the psychosocial experience of dying patients, that is, the 'family support pathway" and the "familial-altruistic pathway." Further reflections on the findings may suggest that these patients may be situated in a "support paradox, "in which they desire family support but also worry about the burden that support places on family members. Implications for practice were discussed in the cultural context. PMID- 19806712 TI - Disability grant for people living with HIV/AIDS in the Eastern Cape of South Africa. AB - A cross-sectional explorative descriptive qualitative/quantitative study was conducted in the Eastern Cape of South Africa. The qualitative component involved 38 HIV/AIDS stakeholders who were interviewed telephonically using a semi structured interview schedule. The quantitative component comprised 607 People Living With HIV/AIDS (PLWHA) who were interviewed using a structured questionnaire. The majority of PLWHA were female, never married, unemployed, aged between 26 to 45 years, Black African, and had more than grade 7 education. All stakeholders supported the disability grant (DG) because it improved the lifestyle of PLWHA. The CD4 count was cited as the main criterion for putting PLWHA on the DG. The conditions and characteristics of the DG were not clearly explained to PLWHA. The DG application process was viewed to be too long. Access to service points was perceived as a challenge for some PLWHA. The DG was used to meet basic household and health care needs. Not being on a DG was associated with lower CD4 counts, often without enough food, and less often without needed medicines in the past 12 months. Having the DG stopped was associated with often not having enough medicines that were needed in the past 12 months. We conclude that the DG is a lifeline for most HIV/AIDS-affected families. We recommend that DG should not use CD4 cell counts as criterion for DG eligibility; the conditions and characteristics of the DG should be fully explained to recipients; the DG application process should be completed within one day; PLWHA who no longer qualify for the DG yet do not have adequate financial means to meet basic necessities should be put on a nutritional support program; and access to the location of the grants by the poor and vulnerable should be improved. PMID- 19806713 TI - Using microbes and wastewater to desalinate water. PMID- 19806714 TI - UV-stabilizing chemicals contaminating Japan's marine environment. PMID- 19806715 TI - Methyl iodide, a fumigant under fire. PMID- 19806716 TI - Landmark global fisheries assessment finds good and bad news. PMID- 19806717 TI - E-waste reuse may be more pervasive than previously thought. PMID- 19806718 TI - Beyond regulation: risk pricing and responsible innovation. PMID- 19806719 TI - Assessment of technologies to meet a low carbon fuel standard. AB - California's low carbon fuel standard (LCFS) was designed to incentivize a diverse array of available strategies for reducing transportation greenhouse gas (GHG) emissions. It provides strong incentives for fuels with lower GHG emissions, while explicitly requiring a 10% reduction in California's transportation fuel GHG intensity by 2020. This paper investigates the potential for cost-effective GHG reductions from electrification and expanded use of biofuels. The analysis indicates that fuel providers could meetthe standard using a portfolio approach that employs both biofuels and electricity, which would reduce the risks and uncertainties associated with the progress of cellulosic and battery technologies, feedstock prices, land availability, and the sustainability of the various compliance approaches. Our analysis is based on the details of California's development of an LCFS; however, this research approach could be generalizable to a national U.S. standard and to similar programs in Europe and Canada. PMID- 19806720 TI - Stable carbon isotope fractionation of 1,2-dichloropropane during dichloroelimination by Dehalococcoides populations. AB - The isotope fractionation of 1,2-dichloropropane (1,2-D) during dichloroelimination to propene by Dehalococcoides populations was explored in laboratory experiments in order to provide data for the characterization of the fate of 1,2-D in heterogeneous subsurface systems. Compound specific stable carbon isotope analysis (CSIA) was used to determine the bulk enrichment factors (epsilonbulk), reactive position specific enrichment factors (epsilonreactive), and apparent kinetic isotope effect (AKIE) values for 1,2-D dichloroelimination in two distinct Dehalococcoides-containing cultures. The epsilonbulk factors calculated in the two cultures were statistically identical, -10.8 +/- 0.9 and 11.3 +/- 0.8 per thousand, even though the cultures were derived from geographically distinct locations. AKIE values for 1,2-D dichloroelimination assuming stepwise and concerted reaction mechanisms were approximately 1.033 and 1.017, respectively. These values are within the range of previously reported values for dichloroelimination reactions and were equivalent to values reported for biotic 1,2-dichloroethane and abiotic 1,1,2,2,-tetrachloroethane and pentachloroethane dichloroelimination reactions. PMID- 19806721 TI - Occurrence and concentrations of benzotriazole UV stabilizers in marine organisms and sediments from the Ariake Sea, Japan. AB - The benzotriazole UV stabilizers, which are used in a variety of plastic products, were analyzed in marine organisms and sediments collected from the Ariake Sea, Japan. The UV stabilizers, such as UV-320, UV-326, UV-327, and UV-328 were detected in all of the samples analyzed, suggesting the production and use of these compounds in Japan. High concentrations of UV stabilizers were found in clams, oysters, and gastropods collected from the tidal flat at concentrations on the order of several hundreds of ng/g on a lipid weight (wt.) basis. The higher trophic species, such as hammerhead sharks and coastal birds, accumulated UV stabilizers, with mean concentrations of 190 ng/g and 74 ng/g (lipid wt.), respectively. These results indicate that benzotriazole UV stabilizers are persistent and bioaccumulative in the marine food-chains. The benzotriazole UV stabilizers were also detected in coastal and river sediments around the Ariake Sea, at concentrations in the range of 7.9-720 ng/g (dry weight basis). Significant correlations were found between concentrations of UV stabilizers and organic carbon content in sediments, implying adsorption of these compounds to organic matter. To our knowledge, this is the first report of ubiquitous contamination and bioaccumulation of benzotriazole UV stabilizers in the marine environment. PMID- 19806722 TI - Photo-Fenton reaction at near neutral pH. AB - The photo-Fenton reaction, oxidation of photoproduced ferrous iron by hydrogen peroxide, produces reactive oxidants that may be important to degradation of biologically and chemically recalcitrant organic compounds in surface waters at circum-neutral pH. Sufficient Fe(II) for the photo-Fenton reaction was produced in situ at two field sites (pH 6.1-7.1) and during irradiations of model systems at pH 7.0 with Suwannee River fulvic acid (SRFA). Amorphous iron oxyhydroxide preparations were much more easily photoreduced than ferrihydrite (Ferr-90). The rate of the photo-Fenton reaction was measured as the difference in the H2O2 accumulation rate in irradiations without and with iron in model systems. Use of benzene as a probe for hydroxyl radical (OH) and nitrate photolysis as the OH source indicated that the yield of phenol from the reaction of benzene with OH is reduced in the presence of iron and SRFA. Even when this reduction in yield was accounted for, the rate of OH production from the Fenton reaction in the model systems was much smaller than the rate of the photo-Fenton reaction. These results indicate that OH is not the only oxidant produced by the photo-Fenton reaction in circum-neutral natural waters; however the photo-Fenton reaction could still be significant for contaminant degradation. PMID- 19806723 TI - Chlorine-36 as a tracer of perchlorate origin. AB - Perchlorate (ClO4(-)) is ubiquitous in the environment. It is produced naturally by atmospheric photochemical reactions, and also is synthesized in large quantities for military, aerospace, and industrial applications. Nitrate-enriched salt deposits of the Atacama Desert (Chile) contain high concentrations of natural ClO4(-), and have been exported worldwide since the mid-1800s for use in agriculture. The widespread introduction of synthetic and agricultural ClO4(-) into the environment has contaminated numerous municipal water supplies. Stable isotope ratio measurements of Cl and O have been applied for discrimination of different ClO4(-) sources in the environment. This study explores the potential of 36Cl measurements for further improving the discrimination of ClO4(-) sources. Groundwater and desert soil samples from the southwestern United States (U.S.) contain ClO4(-) having high 36Cl abundances (36Cl/Cl = 3100 x 10(-15) to 28,800 x 10(-15)), compared with those from the Atacama Desert (36Cl/Cl = 0.9 x 10(-15) to 590 x 10(-15)) and synthetic ClO4(-) reagents and products (36Cl/Cl = 0.0 x 10( 15) to 40 x 10(-15)). In conjunction with stable Cl and O isotope ratios, 36Cl data provide a clear distinction among three principal ClO4(-) source types in the environment of the southwestern U.S. PMID- 19806724 TI - Leachable characteristics of arsenical borogypsum wastes and their potential use in cement production. AB - In this study; the potential use of arsenical borogypsum wastes (ABW) as a set retarder in cement industry was investigated. The comparative performances of arsenical borogypsum wastes (ABW) and natural gypsum samples (NG1 and NG2) at different proportions in the range of 3-8 wt % were tested based on compressive strength over 1, 2, 7, and 28 days and setting times. The use of ABW was observed to lead to a somewhat slower rate of development of strength of the mortar samples than those of NG1 and NG2 during the curing period of 7 days. This is the indication of the effectiveness of ABW as a set retarder. The 28-day compressive strength of mortars tended to decrease with the addition or increasing the proportion of ABW, beyond 5 wt % in particular. The data for setting times of the cement products confirmed set retarding characteristics of ABW with an initial setting time of 90-120 min at 3-5 wt % dosage, which conforms to the desired setting time of > or = 60 min for CEM I (42.5 N) type cement (TS EN 197-1). Leachability tests (TCLP and SPLP) have also shown that ABW can be classified as a nonhazardous waste; but it can readily release metals such as As and Mn, in particular, whereas the mortar samples containing ABW-cement clinker present no environmental concern with its remarkably reduced leachability. PMID- 19806725 TI - Temporal trends of hexabromocyclododecanes (HBCDs) and polybrominated diphenyl ethers (PBDEs) and detection of two novel flame retardants in marine mammals from Hong Kong, South China. AB - Concentrations of hexabromocyclododecanes (HBCDs), polybrominated diphenyl ethers (PBDEs), and three novel flame retardants, namely2-ethylhexyl 2,3,4,5 tetrabromobenzoate (TBB), bis-(2-ethylhexyl)-tetrabromophthalate (TBPH), and hexachlorocyclopentadienyldibromocyclooctane (HCDBCO), were determined in blubber samples of Indo-Pacific humpback dolphins (Sousa chinensis) and finless porpoises (Neophocaena phocaenoides). The levels of HBCDs and PBDEs in cetacean samples ranged from 4.1 to 519 and 103 to 51,100 ng/g lw, respectively. A significant increasing trend of SigmaHBCDs was observed in dolphin samples from 1997 to 2007 with an estimated annual rate of 5%, whereas no significant temporal trends of SigmaPBDEs appeared over the sampling period. This pattern may be attributed to the increasing usage of HBCDs following the restriction/voluntary withdrawal of the production and use of PBDE commercial mixtures in several countries. HCDBCO was not found in the blubber samples. This is the first report of the presence of TBB and TBPH, two new flame retardants that have previously been identified in house dust from the U.S., in marine mammals; concentrations of these compounds in dolphins and porpoises ranged from the instrumental detection limit (IDL) (<0.04) to 70 and IDL (<0.04) to 3859 ng/g lw, respectively. Levels of TBPH were comparable to SigmaHBCDs in porpoise samples. The presence of these novel flame retardants in top-trophic-level marine organisms raises concern about their release into the environment and indicates the need for further monitoring of these compounds in other environmental matrices. PMID- 19806726 TI - Composition of dissolved organic nitrogen in continental precipitation investigated by ultra-high resolution FT-ICR mass spectrometry. AB - The atmospheric transport of fixed nitrogen (N) is a critical component of the global N cycle that has been heavily impacted by human activities. It has been shown that organic N is an important contributor to atmospheric N, but its sources and composition are largely unknown. Rainwater samples collected in New Jersey were analyzed by negative and positive ion ultrahigh-resolution electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Elemental compositions of 402 N-containing compounds were determined and five main groups of compound classes were identified: compounds containing carbon, hydrogen, oxygen, and N detected as positive ions (CHON+), compounds containing CHON detected as negative ions (CHON-), compounds containing CHN detected as positive ions (CHN+), and CHON compounds that contain sulfur (S) detected as both positive and negative ions (CHONS+, CHONS-, respectively). The CHON+ compound class has the largest number of compounds detected (i.e., 281), with the majority, i.e., 207, containing only one N atom. The elemental ratios of these compounds and their detection in the positive ion mode suggest that they are compounds with reduced N functionality. Known contributors to secondary organic aerosol with anthropogenic sources were also identified including organonitrate compounds and nitrooxy organosulfates. PMID- 19806727 TI - Brominated flame retardants, polychlorinated biphenyls, and organochlorine pesticides in bird eggs from the Yellow River Delta, North China. AB - Concentrations of several persistent organohalogen compounds such as dichlorodiphenyltrichloroethane and its metabolites (DDTs), hexachlorocyclohexanes (HCHs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), decabromodiphenylethane (DBDPE), and polybrominated biphenyl 153 (PBB 153) were measured in eggs of six species of wild aquatic birds, one species of wild terrestrial bird, and two species of captive birds from North China. Among the contaminants measured, DDTs were the dominant compounds, HCHs and PCBs were in nearly the same concentration range, and PBDEs exhibited lower concentrations than other compound groups. The median concentrations of DDTs, HCHs, PCBs, and PBDEs in all avian species ranged from 21 to 11034, 5.5 to 623, 1.0 to 613, and 4.6 to 146 ng/g lipid wt, respectively. Median concentrations of DBDPE and PBB 153 in all avian species were in the range of not detectable (ND)-1.7 and ND-0.7 ng/g lipid wt, respectively. Significant differences among species in contaminant profiles and contaminant levels were found depending on their feeding habits, habitat, and migration. The captive birds had the lowest contaminant levels and entirely different congener profiles in PCBs and PBDEs from those of wild birds, which can be attributed to differences in dietary compositions and reproduction rates. Octa- to deca-BDEs contributed more to the total PBDEs in wild terrestrial and captive birds than in wild aquatic birds, except for one insectivorous species, possibly due to greater exposure to terrestrial food sources. Preliminary risk assessment suggests that there is no risk of a reduction in offspring survival in avian species from North China due to organohalogen compounds, except for dichlorodiphenyldichloroethylene (DDE), which would be expected to affect some proportion of the populations of several species of birds studied. PMID- 19806728 TI - Brominated flame retardants in serum from the general population in northern China. AB - In 2006, 128 serum samples were collected from three populations in Tianjin, China: office cleaners, university students, and policemen. These samples were all analyzed for polybrominated diphenyl ethers (PBDEs) and for other brominated flame retardants (BFRs). The median concentration of total PBDEs (sum of 41 congeners) was 7.1 ng/g lipid, ranging from 0.48 to 1980 ng/g lipid. Among these PBDE congeners, the median sum of the tri- to hepta-PBDE (SigmaPBDE3-7) congener concentrations was 2.9 ng/g lipid, ranging from 0.48 to 20 ng/g lipid. The most common tri- to heptabrominated congeners were BDE-47 (30% of total), BDE-99 (24%), BDE-183 (15%), BDE-153 (12%), BDE-28 (9.5%), and BDE-100 (6.2%). These levels of SigmaPBDE3-7 were similar to those observed in Europe and Asia but were much lower than those observed in North America. Highly brominated BDE congeners were detected in some serum samples. In particular, BDE-209 was detected in 28 samples; the median BDE-209 concentration in these samples was 42 ng/g lipid, ranging from ND to 1770 ng/g lipid. The total PBDE levels in office cleaners were significantly higher than in university students and policemen. In addition, we also measured several other BFRs. Hexabromobenzene (HBB) was identified in 26 samples with a median concentration of 0.27 ng/g lipid, ranging from 0.11 to 1.50 ng/g lipid. Pentabromoethylbenzene (PBEB), hexabromocyclododecane (HBCD), 1,2 bis(2,4,6-tribromophenoxy)ethane (BTBPE), and decabromodiphenylethane (DBDPE) were not detected in any of these samples. PMID- 19806729 TI - Partitioning behavior of per- and polyfluoroalkyl compounds between pore water and sediment in two sediment cores from Tokyo Bay, Japan. AB - The partitioning behavior of per- and polyfluoroalkyl compounds (PFCs) between pore water and sediment in two sediment cores collected from Tokyo Bay, Japan, was investigated. In addition, the fluxes and temporal trends in one dated sediment core were studied. Short-chain perfluoroalkyl carboxylic acids (PFCAs) (C < or = 7) were found exclusively in pore water, while long-chain PFCAs (C > or = 11) were found only in sediment The perfluoroalkyl sulfonates (PFSAs), n ethylperfluoro-1-octanesulfonamidoacetic acid (N-EtFOSAA), and perfluorooctane sulfonamide (PFOSA) seemed to bind more strongly to sediment than PFCAs. The enrichment of PFCs on sediment increased with increasing organic matter and decreasing pH. The perfluorocarbon chain length and functional group were identified as the dominating parameters that had an influence on the partitioning behavior of the PFCs in sediment The maximum SigmaPFC contamination in sediment was observed in 2001-2002 to be a flux of 197 pg cm(-2) yr(-1). Statistically significant increased concentrations in Tokyo Bay were found for perfluorooctanesulfonate (PFOS) (1956-2008), perfluorononanoic acid (PFNA) (1990 2008), and perfluoroundecanoic acid (PFUnDA) (1990-2008). Concentrations of PFOSA and N-EtFOSAA increased between 1985 and 2001, but after 2001, the concentration decreased significantly, which corresponded with the phase out of perfluorooctyl sulfonyl fluoride-based compounds by the 3M Company in 2000. PMID- 19806730 TI - Terpenylic acid and related compounds from the oxidation of alpha-pinene: implications for new particle formation and growth above forests. AB - Novel secondary organic aerosol (SOA) products from the monoterpene alpha-pinene with unique dimer-forming properties have been identified as lactone-containing terpenoic acids, i.e., terpenylic and 2-hydroxyterpenylic acid, and diaterpenylic acid acetate. The structural characterizations were based on the synthesis of reference compounds and detailed interpretation of mass spectral data. Terpenylic acid and diaterpenylic acid acetate are early oxidation products generated upon both photooxidation and ozonolysis, while 2-hydroxyterpenylic acid is an abundant SOA tracer in ambient fine aerosol that can be explained by further oxidation of terpenylic acid. Quantum chemical calculations support that noncovalent dimer formation involving double hydrogen bonding interactions between carboxyl groups of the monomers is energetically favorable. The molecular properties allow us to explain initial particle formation in laboratory chamber experiments and are suggested to play a role in new particle formation and growth above forests, a natural phenomenon that has fascinated scientists for more than a century. PMID- 19806731 TI - Graphite- and soot-mediated reduction of 2,4-dinitrotoluene and hexahydro-1,3,5 trinitro-1,3,5-triazine. AB - Black carbon (BC) is an important class of geosorbents that influence the fate and transport of organic pollutants. It is commonly assumed that molecules sorbed to BC are chemically inert. Here we show that this is not true for redox sensitive sorbates such as nitro-aromatic compounds. In the presence of graphite or n-hexane soot as a BC material, the reduction of 2,4-dinitrotoluene (DNT) to 2,4-diaminotoluene by dithiothreitol was greatly accelerated. The para and ortho nitro groups of graphite or soot-sorbed DNT had an approximately equal probability of being reduced. This (1:1) regio-selectivity is different from that when DNT is reduced in homogeneous solution. That is, sorption to BC altered both the kinetics and pathway of DNT reduction. Transformation of hexahydro-1,3,5 trinitro-1,3,5-triazine, a nonaromatic nitro compound, by dithiothreitol was also enhanced by graphite, with concurrent formation of formaldehyde. We propose that BC can catalyze the reduction of nitro compounds because it contains microscopic graphitic (graphene) domains, which are both sorption sites and electron conductors. The environmental significance and potential applications of these findings are discussed. PMID- 19806732 TI - Natural attenuation potential of phenylarsenicals in anoxic groundwaters. AB - The extensive production of chemical warfare agents in the 20th century has led to serious contamination of soil and groundwater with phenyl arsenicals at former ammunition depots or warfare agent production sites worldwide. Most phenyl arsenicals are highly toxic for humans. The microbial degradation of phenylarsonic acid (PAA) and diphenylarsinic acid (DPAA) was investigated in microcosms made of anoxic groundwater/sediment mixtures taken from different depths of an anoxic, phenyl arsenical contaminated aquifer in Central Germany. DPAA was not transformed within 91 days incubation time in any of the microcosms. The removal of PAA can be described by a first order kinetics without a lag-phase (rate: 0.037 d(-1)). In sterilized microcosms, PAA concentrations always remained stable, demonstrating that PAA transformation was a biologically mediated process. PAA transformation occurred under sulfate-reducing conditions due to sulfate consumption and production of sulfide. The addition of lactate (1 mM), a typical substrate of sulfate-reducing bacteria, increased the transformation rate of PAA significantly up to 0.134 d(-1). The content of total arsenic was considerably reduced (> 75%). Intermediates of PAA transformation were detected by high performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS). Experiments with a pure strain and sterile controls of Desulfovibrio gigas spiked with PAA showed that the elimination process is linked to the presence of sulfide formed through bacterial activity. Phenyl arsenicals were likely immobilized in the sedimentthrough sulfur substitution and a subsequent sulfur bond under the prevailing sulfate reducing condition. The results of this study indicate that PAA can undergo microbiologically mediated transformation in anoxic aquifers, leading to reduced concentrations in groundwater, which indicate a (enhancend) natural attenuation potential. PMID- 19806733 TI - Coupled factors influencing concentration-dependent colloid transport and retention in saturated porous media. AB - The coupled influence of input suspension concentration (Ci), ionic strength (IS), and hydrodynamics on the transport and retention of 1.1 microm carboxyl modified latex colloids in saturated quartz sand (150 microm) under unfavorable attachment conditions (pH 10) was investigated. The percentage of retained colloids in column experiments decreased with Ci at intermediate IS conditions (31 or 56 mM) when colloids were weakly associated with the solid phase by a shallow secondary energy minima. In contrast, the effects of Ci on colloid retention were absent when IS was too low (6 mM) or too high (106 mM). The concentration effects under intermediate IS conditions were dependent on the system hydrodynamics, magnitude of Ci, and injection order of Ci, but they were largely independent of the input colloid mass. These observations were explained in part by time- and concentration-dependent filling of retention sites. Only a small fraction of the solid surface area was found to contribute to retention when IS was 31 mM, and micromodel observations indicated that colloid retention was enhanced in lower velocity regions of the pore space that occurred near grain grain contacts. Consequently, retention profiles for IS = 31 mM conditions were increasingly nonexponential at lower values of Ci (during filling), whereas the observed concentration effect was largely eliminated as retention locations became filled. In addition, micromodel observations indicated that liquid and solid phase mass transfer of colloids to retention locations was influenced by Ci under intermediate IS conditions. Higher values of Ci are expected to produce less relative mass transfer to retention locations due to increased numbers of collisions that knock weakly associated colloids off the solid phase. Hence, the concentration effects were found to be largely independent of input colloid mass during filling of retention sites. PMID- 19806734 TI - Formation of iodinated organic compounds by oxidation of iodide-containing waters with manganese dioxide. AB - This study shows that iodinated organic compounds can be produced when iodide containing waters are in contact with manganese oxide birnessite (delta-MnO2) in the pH range of 5-7. In the absence of natural organic matter (NOM), iodide is oxidized to iodate that is also adsorbed onto delta-MnO2. In the presence of iodide and NOM, adsordable organic iodine compounds (AOI) are formed at pH < 7 because of the oxidation of iodide to iodine by delta-MnO2 and the reactions of iodine with NOM. In addition, iodoacetic acid and iodoform have been identified as specific iodinated byproducts. Formation of iodoform is not observed for high NOM/delta-MnO2 ratios due to inhibition of the catalytic effect of delta-MnO2 by NOM poisoning. Experiments with model compounds such as resorcinol and 3,5 heptanedione confirmed that the delta-MnO2/l(-) system is very effective for the formation of iodinated organic compounds. These results suggest that birnessite acts as a catalyst through the oxidation of iodide to iodine and the polarization of the iodine molecule, which then reacts with NOM moieties. Furthermore, our results indicate that during water treatment in the presence of manganese oxide, iodinated organic compounds may be formed, which may lead to taste and odor or toxicological problems. PMID- 19806735 TI - Binding of Hg2+ with phytochelatins: study by differential pulse voltammetry on rotating Au-disk electrode, electrospray ionization mass-spectrometry, and isothermal titration calorimetry. AB - The binding of Hg2+ with synthetic phytochelatins ((gamma-Glu-Cys)n-Gly, PCn, n = 2, 3, 4) was investigated by a recently proposed electroanalytical method, using differential pulse voltammetry on the rotating Au-disk electrode, Electrospray ionization mass-spectrometry (ESI-MS) and isothermaltitration calorimetry (ITC). ESI-MS experiments provided the exact stoichiometries of the complexes formed at different PCn/Hg2+ ratios. Voltammetry provided more detailed information on the complexation processes through the use of multivariate curve resolution by alternating least squares of the data matrix obtained from titrations withfine increments of metal or ligand. The system Hg2+-GSH-PC2 was investigated by voltammetry in order to obtain an estimation of the Hg2+ behavior in the presence of two related ligands. The additional assessment of the stability of Hg2+-PCn complexes was achieved through ITC by using the therapeutic chelator sodium 2,3 dimercaptopropanesulfate (DMPS) over Hg2+-PCn systems. The stability of various Hg2+-PCn complexes and the ability of DMPS to replace PCn from these complexes were examined. PMID- 19806736 TI - Mercury bioaccumulation in a stream network. AB - Mercury (Hg) contamination is common in stream and river ecosystems, but factors mediating Hg cycling in the flowing waters are much less understood than inthe lakes and wetlands. In this study, we examined the spatial patterns of methylmercury (MeHg) concentrations in the dominant groups of aquatic insect larvae across a network of streams (drainage area ranging from 0.5 to 150 km2) in northern California during summer baseflow conditions. We found that, with the exception of water striders, all invertebrate groups showed significant (p < 0.05) increases in MeHg concentrations with drainage area. The largest stream in our study watershed, the South Fork Eel River, had the highest aqueous MeHg concentration (unfiltered: 0.13-0.17 ng L(-1)) while most of the upstream tributaries had aqueous MeHg concentrations close to or below the established detection limits (0.02 ng L(-1)). A filamentous alga abundant in South Fork Eel River (Cladophora glomerata) had an exceptionally high fraction of total-Hg as MeHg (i.e., %MeHg from 50-100%). Since other potential hotspots of in-stream Hg methylation (e.g., surface sediment and deep pools) had %MeHg lower than or similar to surface water (approximately 14%), we hypothesize that Cladophora and possibly other autotrophs may serve as hotspots of in-stream MeHg production in this bedrock-dominated stream. Recent studies in other regions concluded that wetland abundance in the watershed is the predominant factor in governing Hg concentrations of stream biota. However, our results show that in the absence of wetlands, substantial spatial variation of Hg bioaccumulation can arise in stream networks due to the influence of in-stream processes. PMID- 19806737 TI - Ensemble-trained PM2.5 source apportionment approach for health studies. AB - An ensemble-trained chemical mass balance (CMB) approach is developed for particulate matter (PM) source apportionment (SA), particularly for use in health studies. The approach uses results from a short-term emission-based chemical transport model (CTM) and multiple receptor-based approaches. Ensemble results have less day-to-day variation in source impacts and fewer biases between observed and estimated PM2.5 mass compared to the original receptor model results. Ensemble results show increases in road dust, biomass burning, and coal impacts, but secondary organic carbon (SOC) impacts decrease. These results, along with observations, are then used to obtain new source profiles. Two sets of new source profiles based on ensemble results in summer (July 2001 and winter (January 2002) were developed, and used in separate CMB applications for a 12 month data set of daily PM2.5 measurements at the Atlanta, GA, Jefferson Street site. Results show that ensemble-trained CMB approaches, using both summer profiles and winter profiles, effectively reduce day-to-day variability of source impact estimates by reducing fewer days of zero impact from sources known to be present as compared to traditional receptor modeling, suggesting improved results. PMID- 19806738 TI - Multivariate relationships between molecular descriptors and isomer distribution patterns of PCDD/Fs formed during MSW combustion. AB - The isomer distribution patterns of mono- to hepta-chlorinated dibenzo-p-dioxins (PC1-7DD) and dibenzofurans (PC1-7DF) in postcombustion zone flue gas during incineration of an artificial municipal solid waste in a laboratory-scale fluidized-bed reactor were evaluated. Bidirectional orthogonal projections to latent structures (O2PLS) was used to correlate a set of physicochemical properties and chlorine substitution descriptors with the objective to identify parameters correlated with postcombustion zone PCDD and PCDF formation. The most influential variable for the distribution of PCDD congeners was chlorine substitution in positions 1 and 3 (Cl1+3), and overall the chlorine substitution descriptors exerted a larger impact on PCDDs than on PCDFs. For the PCDF, chlorination of the 9-position was the most influential X-variable. Distinct clustering was observed and was most pronounced for PCDFs, dividing mostof the homologues into two or three subgroups of congeners. These subgroups seemed to correspond to the probability of formation by chlorophenol condensation. The sterically crowded dibenzofuran bay-sites (1- and 9-positions) were found to negatively influence PCDF formation, with chlorination of the 9-position having the greatest impact. Since PCDD/F toxicity is related to the lateral positions, elucidating the factors governing chlorination may be of great importance for detoxification of incineration byproducts. PMID- 19806739 TI - Identification of transformation products of organic contaminants in natural waters by computer-aided prediction and high-resolution mass spectrometry. AB - Transformation products (TPs) of organic contaminants in aquatic environments are still rarely considered in water quality and chemical risk assessment, although they have been found in concentrations that are of concern. Since many different TPs can potentially be formed in the environment and analytical standards are typically lacking for these compounds, knowledge on the prevalence of TPs in aquatic environments is fragmentary. In this study, an efficient procedure was therefore developed to comprehensively screen for large numbers of potential TPs in environmental samples. It is based on a target list of plausible TPs that has been assembled using the University of Minnesota Pathway Prediction System (UM PPS) for the computer-aided prediction of products of microbial metabolism and an extensive search for TPs reported in the scientific literature. The analytical procedure for screening of the compounds on the target list has been developed to allow for the detection of a broad range of compounds in complex environmental samples in the absence of commercially available reference standards. It includes solid phase extraction with broad enrichment efficiency, followed by liquid chromatography and tandem mass spectrometry with high mass resolution and accuracy. The identification of target TPs consisted of extracting the exact mass from the chromatogram, selecting peaks of sufficient intensity, checking the plausibility of the retention time, and interpreting mass spectra. The procedure was used to screen for TPs of 52 pesticides, biocides, and pharmaceuticals in seven representative surface water samples from different regions in Switzerland. Altogether, 19 TPs were identified, including both some well-known and commonly detected TPs, and some rarely reported ones (e.g., biotransformation products of the pharmaceuticals venlafaxine and verapamil, or of the pesticide azoxystrobin). Overall, the rather low number of TPs detected suggests that TPs may not pose a problem of unexpected magnitude for aquatic resources. PMID- 19806740 TI - Polymer-water partition coefficients of hydrophobic compounds for passive sampling: application of cosolvent models for validation. AB - Polymer-water partition coefficients (Kpw) of hexachlorobenzene, 41 polychlorinated biphenyls (PCBs), and 26 polyaromatic hydrocarbons (PAHs) were determined for low-density polyethylene (LDPE) and five different silicone rubbers. Partition coefficients were determined in ultra pure water and in a range of methanol-water mixtures. Different cosolvent models for the effect of methanol concentration on the polymer-mixture partition coefficient (Kpm) were used to validate the Kpw in pure water. Linear regression of logKpm against the mole fraction (x) methanol over range 0 < x < 0.3 yielded the best results. The obtained logKpws were best described by a correlation with molecular weight, for PCBs in combination with the fraction of chlorine atoms in the meta and para positions (standard deviations of approximately 0.08 log units). Correlations with logKow were less good (standard deviations of approximately 0.21 log units), partly as a result of uncertainties in the logKow estimates that were used. Similar Kpws were found for different batches of silicone rubber from the same supplier. Differences in logKpws for silicone rubbers obtained from different suppliers ranged from 0.16-0.58. PMID- 19806741 TI - Comprehensive non-targeted analysis of contaminated groundwater of a former ammunition destruction site using 1H-NMR and HPLC-SPE-NMR/TOF-MS. AB - The aim of the present study was to explore the capabilities of the combination of 1H NMR (proton nuclear magnetic resonance) mixture analysis and HPLC-SPE NMR/TOF-MS (high-performance liquid chromatography coupled to solid-phase extraction and nuclear magnetic resonance and time-of-flight mass spectrometry) for the characterization of xenobiotic contaminants in groundwater samples. As an example, solid-phase extracts of two groundwater samples taken from a former ammunition destruction site in Switzerland were investigated. 1H NMR spectra of postcolumn SPE enriched compounds, together with accurate mass measurements, allowed the structural elucidation of unknowns. This untargeted approach allowed us to identify expected residues of explosives such as 2,4,6-trinitrotoluene (2,4,6-TNT), Hexogen (RDX) and Octogen (HMX), degradation products of TNT (1,3,5 trinitrobenzene (1,3,5-TNB), 2-amino-4,6-dinitrotoluene (2-A-4,6-DNT), 3,5 dinitrophenol (3,5-DNP), 3,5-dinitroaniline (3,5-DNA), 2,6-dinitroanthranite, and 2-Hydroxy-4,6-dinitrobenzonitrile), benzoic acid, Bisphenol A (a known endocrine disruptor compound), and some toxicologically relevant additives for propelling charges: Centralite I (1,3-diethyl-1,3-diphenylurea), DPU (N,N-diphenylurethane), N,N-diphenylcarbamate (Acardite II), and N-methyl-N-phenylurethane. To our knowledge, this is the first report of the presence of these additives in environmental samples. Extraction recoveries for Centralite I and DPU have been determined. Contaminants identified by our techniques were quantified based on HPLC-UV (HPLC-ultraviolet detection) and 1H NMR mixture analysis. The concentrations of the contaminants ranged between 0.1 and 48 microg/L assuming 100% recovery for the SPE step. PMID- 19806742 TI - Removal of acetaminophen using enzyme-mediated oxidative coupling processes: I. Reaction rates and pathways. AB - We found that acetaminophen could be effectively transformed and removed from water by laccase-mediated oxidative coupling processes. The removal of acetaminophen followed second-order kinetics with first-order to the concentrations of both the substrate and the enzyme. Mass spectrum analysis demonstrated that polymerization through radical-radical coupling mechanism was the pathway leading to acetaminophen transformation. Coupling products thus formed are believed to be biologically inactive and more readily removable from water. Secondary mass spectra of the dimers in combination with molecular modeling analysis further elucidated that the coupling proceeded via covalent bonding between two molecules at their unsubstituted carbons in benzene rings. These findings demonstrated that laccase-mediated oxidative coupling can potentially serve as an alternative strategy to control certain micropollutants in water/wastewater treatment and reuse. PMID- 19806743 TI - Removal of acetaminophen using enzyme-mediated oxidative coupling processes: II. Cross-coupling with natural organic matter. AB - The influence of natural organic matter (NOM) on the transformation of acetaminophen in laccase-mediated oxidative coupling systems was investigated in this study. It was found that the removal of acetaminophen was enhanced while the self-coupling of acetaminophen was suppressed in the presence of dissolved NOM, likely resulting from cross-coupling between dissolved NOM and acetaminophen. In additionto cross-coupling with acetaminophen, NOM moieties could couple to each other upon reaction with laccase. This was evidenced by the development of a characteristic absorbance band centered at 472 nm. According to the rate of the absorbance change at 472 nm, the NOM coupling reactions in four different NOM solutions were evaluated. Apparently, the tendency of NOM coupling reactions correlates with the tendency of acetaminophen cross coupling with NOM in these solutions. Possible reaction pathways of cross-coupling were explored using guaiacol as a model NOM proxy, and the products were extracted and analyzed with mass spectrometry (MS). The results suggested that acetaminophen and guaiacol molecules were cross-coupled via the formation of C-O-C bonds. PMID- 19806744 TI - Separating and recycling metals from mixed metallic particles of crushed electronic wastes by vacuum metallurgy. AB - During the treatment of electronic wastes, a crushing process is usually used to strip metals from various base plates. Several methods have been applied to separate metals from nonmetals. However, mixed metallic particles obtained from these processes are still a mixture of various metals, including some toxic heavy metals such as lead and cadmium. With emphasis on recovering copper and other precious metals, there have hitherto been no satisfactory methods to recover these toxic metals. In this paper, the criterion of separating metals from mixed metallic particles by vacuum metallurgy is built. The results show that the metals with high vapor pressure have been almost recovered completely, leading to a considerable reduction of environmental pollution. In addition, the purity of copper in mixed particles has been improved from about 80 wt % to over 98 wt %. PMID- 19806745 TI - A new visible-light photocatalyst: CdS quantum dots embedded mesoporous TiO2. AB - Cadmium sulfide quantum dots (QDs) sensitized mesoporous TiO2 photocatalysts were prepared by preplanting cadmium oxide as crystal seeds into the framework of ordered mesoporous titanium dioxide and then converting CdO to CdS QDs through ion-exchange. The presence of CdS QDs in the TiO2 framework extended its photoresponse to the visible-light region by accelerating the photogenerated electron transfer from the inorganic sensitizer to TiO2. The new photocatalyst showed excellent photocatalytic efficiency for both oxidation of NO gas in air and degradation of organic compounds in aqueous solution under visible light irradiation. The photocatalysts were characterized byX RD, N2 adsorption desorption, TEM, XPS, UV/vis, and PL spectroscopy. The relationship between the physicochemical properties and the photocatalytic performance of the sample is discussed. PMID- 19806746 TI - Managing pore-water quality in mine tailings by inducing microbial sulfate reduction. AB - A field-scale experiment was conducted to evaluate the potential for inducing microbial sulfate reduction as a passive in situ technique for managing water quality in mine tailings deposits. Sulfide- and carbonate-rich minetailings, characterized by near-neutral pH pore water, were amended with < 1 dry wt. % organic carbon. The geochemical evolution of pore water was monitored for four years. The results demonstrate that organic carbon supported dissimilatory sulfate reduction (DSR) in the vadose zone. Decreases in dissolved SO4 and S2O3 were accompanied by H2S production, increased populations of sulfate-reducing bacteria (SRB), 34S-SO4 enrichment, and undersaturation of pore water with respect to gypsum [CaSO4 x 2H2O]. The mass of dissolved S decreased by > 45% during the monitoring period, which coincided with the removal of Zn, Sb, and Tl. Mobilization of Fe and As occurred initially; however, subsequent decreases in aqueous concentrations were observed. Mineralogical investigation confirmed the presence of secondary Fe-S and Zn-Fe-S phases. Amendment of tailings with a small and dispersed mass of organic carbon resulted in a general decrease in mass transport of sulfide oxidation products. PMID- 19806747 TI - Emissions of inorganic and organic arsenic compounds via the leachate pathway from pretreated municipal waste materials: a landfill reactor study. AB - The emission of arsenic (As) with leachate from mechanically biologically pretreated municipal solid waste (MBP-MSW) was quantified over one year using landfill simulation reactors. Arsenic mobilization and transformation processes were studied by simulating different environmental conditions (anoxic conditions with underlying soil or oxic/anoxic conditions). Amounts of mono-, di-, and trimethylated As in MBP-MSW prior to simulation were < 48 microg As kg(-1) and were magnified to 300-390 microg As kg(-1) under anoxic conditions, whereas methylated As was undetectable in the oxic setup. The highest leachate concentrations (up to 84 microg L(-1)) occurred during the first four weeks of manipulation. The annual Astotal release with leachates averaged 19.6, 7.6, and 4.5 microg kg(-1) under an anoxic environment with underlying soil, oxic conditions, and anoxic conditions, respectively, with 15-50% occurring as organic As. The annually released As represented 0.2-0.8% of the Astotal pool, suggesting that As mobilization from waste is a slow process. The anoxia diminished As release rates, whereas anoxic conditions with underlying soil material elevated the As mobilization, probably due to reductive dissolution of soil-derived Fe and Mn (hydr)oxides. The mass balance of methylated As in MBP-MSW and leachates before and after the treatments highlights As methylation under anoxic conditions and demethylation under oxic landfill conditions. PMID- 19806748 TI - Application of electrolysis to stimulate microbial reductive PCE dechlorination and oxidative VC biodegradation. AB - A novel approach was applied to stimulate biodegradation of chloroethenes bya coupled bioelectro-process. In a flow-through column system, microbial dechlorination of tetrachloroethene to cis-dichloroethene, vinyl chloride, and ethene was stimulated by hydrogen produced by water electrolysis. Dechlorinating bacteria (Dehalococcoides spp. and Desulfitobacterium spp.) and also methanogens and homoacetogens were detected in the anaerobic column. Simultaneously, oxidative biodegradation of lower chlorinated metabolites (vinyl chloride) was stimulated by electrolytic oxygen formation in the corresponding aerobic column. The process was stable for more than 100 days and an average removal of approximately 23 micromol/d PCE and 72 micromo/d vinyl chloride was obtained with a current density of 0.05 mA/cm2. Abiotic electrochemical degradation of the contaminants was not observed. Microbial dechlorination correlated with the current densities in the applied range of 0.01-0.05 mA/cm2. The results are promising for environmental applications, since with electrolysis hydrogen and oxygen can be supplied continuously to chloroethene degrading microorganisms, and the supply rates can be easily controlled by adjusting the electric current. PMID- 19806749 TI - Biodegradation of 4-nitrophenol in a two-phase system operating with polymers as the partitioning phase. AB - The present study has demonstrated the enhanced performance of a two-phase bioreactor, operating with polymers as a partitioning phase, as an alternative to both single phase biotreatment and to the use of an immiscible organic solvent partitioning phase, to deliver a toxic substrate (4-nitrophenol, or 4NP) to a microbial consortium in batch and repeated batch mode. Three commercial polymers were tested, Hytrel, Tone, and Elvax, and were shown to have superior properties related to the use of a consortium, including complete biocompatibility with the biomass and nonbiodegradability. Repeated kinetic tests performed with short reaction times demonstrated the accumulation of 4NP within the polymers in the range of 6-8 mg/g polymer, which reduced polymer performance in subsequent batch operations. Hytrel gave the best performance with residuals of up to 4 mg/g polymer showing no reduction in subsequent use, while for the other polymers a 4NP value lower than 2 mg/g polymer was required to have acceptable performance during repeated polymer use. Polymer reuse without affecting the process efficiency was confirmed with regeneration tests. A conventional methanol extraction method, as well as biological regeneration of the polymers by prolonged contact with the biomass, were assessed for their ability to remove the residual 4NP. Parallel kinetic tests performed with newand regenerated polymers showed a complete overlap of the 4NP concentration profiles indicating that a simple biological regeneration method provides a means of completely restoring polymer performance for repeated batch operation. PMID- 19806750 TI - Estrogen biodegradation kinetics and estrogenic activity reduction for two biological wastewater treatment methods. AB - Estrogens from anthropogenic and livestock sources are a serious concern for aquatic ecosystems at concentrations less than 1 ng/L Fundamental process parameters to reduce estrogenic activity were investigated for two biotreatment methods: heterotrophic bacterial degradation in municipal activated sludge (AS) and a nitration process that is applicable to high NH4-N wastewaters. Batch tests with estrogen and nitro-estrogen compounds were conducted at nanogram per liter concentrations with mixed liquor from an AS wastewater treatment facility (WWTF) operating at a 3 day solids retention time (SRT) and a membrane bioreactor (MBR) WWTF operating at a 30-40 day SRT. The estrogenic activities of estrone (E1), 17beta-estradiol (E2), and 17alpha-ethinylestradiol (EE2) were reduced 80-97% following nitration. First-order biological degradation rate coefficients (kb) of the nitrated estrogens were 10-50% lower than the parent estrogen compounds. The kb values for EE2 in MBR and AS mixed liquors were similar, 1.67 and 1.63 L/gVSS day respectively, indicating that the bacteria responsible for EE2 degradation were present at long and short SRTs. The kb values for E1 and E2 were 2 orders of magnitude greater than for EE2. EE2 degradation was 7.5 times faster in the presence of E1 and E2, and no effect was observed with other estrogen mixtures. PMID- 19806751 TI - CaO-based pellets supported by calcium aluminate cements for high-temperature CO2 capture. AB - The development of highly efficient CaO-based pellet sorbents, using inexpensive raw materials (limestones) or the spent sorbent from CO2 capture cycles, and commercially available calcium aluminate cements (CA-14, CA-25, Secar 51, and Secar 80), is described here. The pellets were prepared using untreated powdered limestones or their corresponding hydrated limes and were tested for their CO2 capture carrying capacities for 30 carbonation/calcination cycles in a thermogravimetric analyzer (TGA). Their morphology was also investigated by scanning electron microscopy (SEM) and their compositions before and after carbonation/calcination cycleswere determined by X-ray diffraction (XRD). Pellets prepared in this manner showed superior behavior during CO2 capture cycles compared to natural sorbents, with the highest conversions being > 50% after 30 cycles. This improved performance was attributed to the resulting substructure of the sorbent particles, i.e., a porous structure with nanoparticles incorporated. During carbonation/calcination cycles mayenite (Ca12Al14O33) was formed, which is believed to be responsible for the favorable performance of synthetic CaO-based sorbents doped with alumina compounds. An added advantage of the pellets produced here is their superior strength, offering the possibility of using them in fluidized bed combustion (FBC) systems with minimal sorbent loss due to attrition. PMID- 19806752 TI - Structural concretes with waste-based lightweight aggregates: from landfill to engineered materials. AB - This research provides possible opportunities in the reuse of waste and particularly muds, coming from both ornamental stone (granite sludges from sawing and polishing operations) and ceramic production (porcelain stoneware tile polishing sludge), for the manufacture of lightweight aggregates. Lab simulation of the manufacturing cycle was performed by pelletizing and firing the waste mixes in a rotative furnace up to 1300 degrees C, and determining composition and physicomechanical properties of lightweight aggregates. The best formulation was used to produce and test lightweight structural concretes according to standard procedures. Both granite and porcelain stoneware polishing sludges exhibit a suitable firing behavior due to the occurrence of SiC (an abrasive component) which, by decomposing at high temperature with gas release, acts as a bloating promoter, resulting in aggregates with particle density < 1 Mg/m3. However, slight variations of mixture composition produce aggregates with rather different properties, going from values close to those of typical commercial expanded clays (particle density 0.68 Mg/m3; strength of particle 1.2 MPa) to products with high mechanical features (particle density 1.25 Mg/m3; strength of particle 6.9 MPa). The best formulation (50 wt.% porcelain stoneware polishing sludge +50 wt.% granite sawing sludge) was used to successfully manufacture lightweight structural concretes with suitable properties (compressive strength 28 days > 20 MPa, bulk density 1.4-2.0 Mg/m3). PMID- 19806753 TI - Zero-valent aluminum for oxidative degradation of aqueous organic pollutants. AB - Oxidative degradation of aquatic organic contaminants using zero-valent aluminum (ZVAl) in the presence of dissolved oxygen (O2) was investigated. The metal corrosion process in acidic conditions (pH < 4) was accompanied by electron transfer from ZVAl to O2, which led to the simultaneous generation of Al3+ and hydrogen peroxide (H2O2). The oxidation of 4-chlorophenol (4-CP), a model substrate, was initiated by the generation of hydroxyl radicals (HO*) via electron transfer from Al0 to H2O2. Degradation was initiated after an induction period of about 2 h, during which the native oxide layer was dissoluted. The HO*( ) mediated oxidation reaction was completely quenched by adding methanol as a radical scavenger. Systematic studies on the effects of ZVAl loading, pH, and surface oxide content revealed that the oxide layer dissolution controlled the Al0-mediated oxidation of 4-CP. The proposed process is similarly compared with the zero-valent iron (ZVI) system, but the ZVAl/O2 system showed a higher oxidation capacity compared with ZVI/O2 because of the stability of aquo complexed Al3+ ions over a wider pH range. The degradation of phenol, nitrobenzene, and dichloroacetate was also successfully achieved with ZVAl. The present study proposes the ZVAl/O2 process as a viable method of oxidative water treatment. PMID- 19806754 TI - Fullerene exposures with oysters: embryonic, adult, and cellular responses. AB - Oysters are an ecologically important group of filter-feeders, and a valuable toxicology model for characterizing the potential impacts of nanoparticles to marine organisms. Fullerene (C60) exposure studies with oysters, Crassostrea virginica, were conducted with a variety of biological levels, e.g., developmental studies with embryos, whole organism exposures with adults, and isolated hepatopancreas cells. Significant effects on embryonic development and lysosomal destabilization were observed at concentrations as low as 10 ppb. Moreover, based on our extensive experience with the lysosomal assay, the lysosomal destabilization rates at fullerene concentrations > or = 100 ppb were regarded as biologically significant as they are associated with reproductive failure. Interestingly, there was no significant increase in lipid peroxidation levels in hepatopancreas tissues. Oyster hepatopancreas tissues are composed of lysosomal rich cells, and confocal microscopy studies indicated thatthe fullerene particles readily accumulated inside hepatopancreas cells within 4 h. Fullerene aggregates tended to be localized and concentrated into lysosomes. The microscopic work in conjunction with the lysosomal function assays supports the premise that endocytotic and lysosomal pathways may be major targets of fullerenes and other nanoparticles. Nanoparticles that affect normal lysosomal and autophagic processes may contribute to long-term, chronic problems for individual health as well as ecosystem health. PMID- 19806755 TI - Accounting for time-dependent effects in biofuel life cycle greenhouse gas emissions calculations. AB - This paper proposes a time correction factor (TCF) to properly account for the timing of land use change-derived greenhouse gas emissions in the biofuels life cycle. Land use change emissions occur at the outset of biofuel feedstock production, and are typically amortized over an assumed time horizon to assign the burdens of land use change to multiple generations of feedstock crops. Greenhouse gas intensity calculations amortize emissions by dividing them equally over a time horizon, overlooking the fact that the effect of a greenhouse gas increases with the time it remains in the atmosphere. The TCF is calculated based on the relative climate change effect of an emission occurring at the outset of biofuel feedstock cultivation versus one amortized over a time horizon. For time horizons between 10 and 50 years, the TCF varies between 1.7 and 1.8 for carbon dioxide emissions, indicating that the actual climate change effect of an emission is 70-80% higher than the effect of its amortized values. The TCF has broad relevance for correcting the treatment of emissions timing in other life cycle assessment applications, such as emissions from capital investments for production systems or manufacturing emissions for renewable energy technologies. PMID- 19806756 TI - A new method for water desalination using microbial desalination cells. AB - Current water desalination techniques are energy intensive and some use membranes operated at high pressures. It is shown here that water desalination can be accomplished without electrical energy input or high water pressure by using a source of organic matter as the fuel to desalinate water. A microbial fuel cell was modified by placing two membranes between the anode and cathode, creating a middle chamber for water desalination between the membranes. An anion exchange membrane was placed adjacent to the anode, and a cation exchange membrane was positioned next to the cathode. When current was produced by bacteria on the anode, ionic species in the middle chamber were transferred into the two electrode chambers, desalinating the water in the middle chamber. Proof-of concept experiments for this approach, using what we call a microbial desalination cell (MDC), was demonstrated using water at different initial salt concentrations (5, 20, and 35 g/L) with acetate used as the substrate for the bacteria. The MDC produced a maximum of 2 W/m2 (31 W/m3) while at the same time removing about 90% of the salt in a single desalination cycle. As the salt was removed from the middle chamber the ohmic resistance of the MDC (measured using electrochemical impedance spectroscopy) increased from 25 Omega to 970 Omega at the end of the cycle. This increased resistance was reflected by a continuous decrease in the voltage produced over the cycle. These results demonstrate for the first time the possibility for a new method for water desalination and power production that uses only a source of biodegradable organic matter and bacteria. PMID- 19806758 TI - Teach your patients well. PMID- 19806757 TI - Comment on "Comparative assessment of the global fate and transport pathways of long-chain perfluorocarboxylic acids (PFCAs) and perfluorocarboxylates (PFCs) emitted from direct sources". PMID- 19806759 TI - Every physician should have a migraine. PMID- 19806760 TI - A complex constellation of interrelated conditions. PMID- 19806761 TI - Stark law covers a narrow corridor of actions. PMID- 19806762 TI - Normative slogging. PMID- 19806763 TI - The American right to health. PMID- 19806764 TI - And she's not only merely dead, she's really most sincerely dead. PMID- 19806765 TI - And she's not only merely dead, she's really most sincerely dead. PMID- 19806766 TI - And she's not only merely dead, she's really most sincerely dead. PMID- 19806767 TI - Rethinking the ethics of vital organ donations. PMID- 19806768 TI - Quiet moments. PMID- 19806769 TI - Swine flu vaccine: what is fair? PMID- 19806770 TI - Slipping through the net: social vulnerability in pandemic planning. PMID- 19806771 TI - Case study. Dirty blood. PMID- 19806772 TI - Case study. Dirty blood. Commentary. PMID- 19806773 TI - Case study. Dirty blood. Commentary. PMID- 19806774 TI - Toward a meaningful alternative medicine. PMID- 19806775 TI - Misgivings. PMID- 19806776 TI - Government-scripted consent: when medical ethics and law collide. PMID- 19806778 TI - Bioethics & human rights: access to health-related goods. AB - There are many good reasons for a merger between bioethics and human rights. First, though, significant philosophical groundwork must be done to clarify what a human right to health would be and--if we accept that it exists--exactly how it might influence the practical decisions we face about who gets what in very different contexts. PMID- 19806777 TI - Saturday morning in the clinic. PMID- 19806779 TI - Autonomy, beneficence, and gezelligheid: lessons in moral theory from the Dutch. AB - American bioethicists lack the theoretical resources to work in cross-cultural settings. All we have are two approaches to ethics--principles vs. narratives- that are mostly at odds, and neither of which is up to the job. If moral principles are too abstract to be useful, and if stories cannot provide moral authority, then where do we find our moral norms? PMID- 19806780 TI - When it comes to HIV infection, some are more equal than others. PMID- 19806781 TI - Chemopreventive effect of nonsteroidal anti-inflammatory drugs on 9,10 dimethylbenz[a]anthracene-induced lung carcinogenesis in mice. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin, celecoxib, and etoricoxib were studied as chemopreventive agents in lung cancer in mice induced by 9,10-dimethylbenz[a]anthracene (DMBA). The animals were subjected to a single intratracheal instillation of DMBA by surgical intervention, while they were treated with oral NSAIDs daily at their following anti-inflammatory dose: aspirin 25 mg/kg, celicoxib 6 mg/kg, and etoricoxib 0.6 mg/kg body weight, respectively. The animals were sacrificed after 18 weeks of treatment. Results showed a significant incidence of pulmonary tumors, dysplastic changes in histopathology, and signs of inflammatory occurrence in the DMBA-treated animals, which were grossly reversed by the NSAIDs. A greater number of macrophages, neutrophils, and lymphocytes were seen in the bronchoalveolar lavage (BAL) smear while the inflammatory cell counts decreased in DMBA + NSAIDs groups. A significant increase in the drug-metabolizing enzymes viz. cytochrome p450, cytochrome b5, and glutathione-S-transferase was noted in the DMBA group, which was reverted back in the NSAID-treated mice. Similarly, the subcelluler enzymes were elevated in DMBA, but significantly fell in the NSAID groups. DMBA also caused a higher level of lipid peroxidation as well as the different antioxidant enzyme activity, which were corrected by the NSAIDs. A marked elevation was noticed in the total lipid composition and its individual constituents in the DMBA group, which was reverted back appreciably by the NSAIDs. The results suggest that the DMBA induced lung tumor development in balb/c mice could be a reliable model to test the chemopreventive potential of the NSAIDs. PMID- 19806782 TI - Angiogenic pathway inhibition of Corydalis yanhusuo and berberine in human umbilical vein endothelial cells. AB - Corydalis yanhusuo, a well-known traditional Chinese medicine, is widely used in China as an analgesic for patients with terminal cancer. In this study, we want to expose the antiangiogenic effects and the underlying mechanisms of C. yanhusuo and the alkaloids obtained from this plant. The constituents of C. yanhusuo were first investigated for their inhibitory effects on angiogenesis, using several bioassays, including vascular endothelial growth factor (VEGF)-induced human umbilical vein endothelial cell (HUVEC) proliferation, migration, invasion, and tube formation. To determine the active antiangiogenic compounds in C. yanhusuo, we studied the antiproliferative activities of several main constituents of C. yanhusuo, which belong to a group of protoberberine alkaloids, on HUVECs and identified berberine as a powerful angiogenesis inhibitor in C. yanhusuo. Both C. yanhusuo extract and its active compound berberine significantly suppressed the VEGF-induced upregulation of matrix metalloproteinase 2 (MMP2) at both mRNA and protein levels. Their functional effects, including the inhibition of MMP2, were shown to be involved VEGF-triggered ERK1/2 pathways. Our findings provide novel insights into the antiangiogenic effects of C. yanhusuo and berberine, and offer scientific evidence for their traditional clinical application as a cancer treatment. PMID- 19806783 TI - Effects of alpha-adrenoceptor antagonist doxazosin on MDR1-mediated multidrug resistance and transcellular transport. AB - The purpose of this study is to examine the effects of doxazosin, an alpha adrenoceptor antagonist, on P-glycoprotein/MDR1-mediated multidrug resistance (MDR) and the transport of anticancer drugs. The effects of doxazosin, prazosin, and terazosin on MDR1-mediated MDR were assessed in human cervical carcinoma HeLa cells and the MDR1-overexpressing derivative Hvrl00-6, established by stepwise increases of the vinblastine concentration in the culture medium. The effects of doxazosin on the transcellular transport and intracellular accumulation of [3H]vinblastine, [3H]daunorubicin, and [3H]digoxin, all MDR1 substrates, were evaluated using LLC-GA5-COL150 cell monolayers, established by transfection of human MDR1 cDNA into porcine kidney epithelial LLC-PK1 cells. The sensitivity to vinblastine and paclitaxel of Hvrl00-6 cells was increased at 3.4- and 17.5-fold, respectively, by the addition of 1 microM doxazosin, whereas prazosin and terazosin had weaker or no such effects. Prazosin at 1 microM had a reversal effect on the sensitivity to vinblastine, whereas terazosin had no effect. In transport experiments, doxazosin concentration dependently increased the apical to-basal transport of radiolabeled drugs in LLC-GA5-COL150 cells, but did not show remarkable effects on the basal-to-apical transport. In addition, doxazosin restored the intracellular accumulation in a concentration-dependent manner in LLC-GA5-COL150 cells. Doxazosin may partly reverse MDR by inhibiting MDR1 mediated transport, making it a candidate lead compound in the development of a reversing agent for MDR. PMID- 19806784 TI - Hemin counteracts the repression of Bcl-2 and NrF2 genes and the cell killing induced by imatinib in human Bcr-Abl(+) CML cells. AB - Imatinib is a targeted selective inhibitor of chimaeric Bcr-Abl tyrosine kinase developed for effective therapy of chronic myelogenous leukemia (CML) and acute lymphocytic leukemia (ALL) patients. Unfortunately, evidence now exists to indicate that a portion of such patients treated with imatinib acquire resistance and subsequently relapse. To understand the heterogeneous basis of imatinib resistance, we have investigated the possible mechanism(s) via which hemin, a key regulator of hematopoiesis that is converted to heme intracellularly, renders CML cells less susceptible to imatinib. Hemin at 30-90 aM protected a substantial proportion (>40%) of human Bcr-Abl(+) CML cells (K-562 and KU-812) from imatinib induced cell killing by increasing the imatinib IC50 value, reducing DNA damage, and promoting erythroid differentiation. RT-PCR assessment of RNA transcripts encoded by human GAPDH, Ggamma-globin, Bcr-Abl, HO-2, Hpr-6, CEBPa, Bcl-2a, Bcl 2b, and Nrf2 genes revealed that hemin selectively counteracted the repression of antiapoptotic Bcl-2a, Bcl-2b, and Nrf2 genes in imatinib-treated cells. These genes are markedly repressed by imatinib alone in human K-562 CML cells. Hemin, however, had no detectable effect on the expression of the Bcr-Abl gene. Moreover, inhibition of de novo heme biosynthesis by succinyl-acetone enhanced the killing effect of imatinib. These data clearly indicate that: (a) cellular heme resulted from de novo biosynthesis and hemin uptake alters the developmental stage of human Bcr-Abl(+) CML cells and their susceptibility to imatinib; (b) cellular heme counteracts the ability of imatinib to repress Bcl-2 and Nrf2 gene expression; and (c) inhibitors of de novo biosynthesis can be developed and combined with imatinib to enhance its antileukemic activity. PMID- 19806785 TI - Evaluation of global genome methylation in gastritis lesion and its correlation with clinicopatological findings. AB - Global genome hypomethylation as an epigenetic phenomenon may induce (pre)neoplastic transformation through inducing chromosomal and genomic instability and activating oncogenes. Global genome hypomethylation has a fundamental role in early stages of tumorigenesis but little is known about this epigenetic event in gastric precancerous lesions such as gastritis. Therefore, we decided to evaluate this issue in gastritis lesion for obtaining new insight toward molecular biology of gastric cancer. Here we used a technique composed of restriction enzyme digestion and pyrosequencing known as luminometric methylation assay to evaluate this issue. DNA obtained from normal and gastritis lesions was digested with HpaII (sensitive to methylation in its cut site) and MspI (insensitive). Overhangs resulting from these enzymes then fill in by polymerase extension assay using pyrosequencing instrument. Nucleotide incorporation during polymerase extension generates light, which expresses as pick in the pyrogram. By comparing the height of picks obtained form both enzymes it can be possible to evaluate and compare global genome methylation level of gastritis and normal tissues. If the target site is fully methylated, the HpaII/MspI (their pick height) will approach zero. If not, this ratio will be around 1. In the other conditions this ratio varies between 0 and 1. Comparing the ratio of normal and gastritis sample, it can be inferred whether or not gastritis is hypomethylated. This study was performed on 83 gastritis and normal adjacent tissues. The patients included 34 male and 49 female and were 15 to 83 years old. According to our study, gastritis tissue was hypomethylated more than the normal tissue (p = 0.028). Global genome methylation has no significant correlation with MSI, pathological findings, age, and gender. We conclude that global genome hypomethylation occurs in the gastritis level. This reduction probably continues in the next steps toward gastric cancer and may induce other epigenetic and/or genetic changes (such as MSI) that promote carcinogenesis. PMID- 19806786 TI - Gemcitabine plus irinotecan as first-line weekly therapy in locally advanced and/or metastatic pancreatic cancer. AB - Single-agent gemcitabine has been established as standard treatment for advanced pancreatic cancer since clinical studies have shown an improvement in overall survival and significant clinical benefit when compared to the best supportive care despite low overall objective response. Several phase II studies have tested other single agents and different gemcitabine-based regimens in pancreatic cancer, but both response and survival rates have remained low. Irinotecan, a topoisomerase I inhibitor currently approved for the treatment of metastatic colon cancer, has also demonstrated improved response rate in patients with pancreatic cancer. Our purpose was to determine the activity and toxicity of this regimen in patients with unresectable or metastatic pancreatic cancer. Patients with histologically confirmed pancreatic adenocarcinoma received gemcitabine 1000 mg/m2 plus irinotecan 100 mg/m2 IV on days 1, 8, and 15 of a 28-day cycle for 6-8 months. From February 2004 to April 2006, 33 patients were entered into this study, 32 of whom were evaluable for treatment response, toxicity, median time to progression, and median survival. Characteristics included a median age of 63 years (range 41-79), 21 males (64%), and 12 females (36%). One patient discontinued treatment due to adverse effects. The total number of cycles administered was 188 and the median number of cycles for patients was 5.6 (range 2-7). Thirty-two patients were assessable for toxicity and response. Grade 3 hematological toxicity occurred in 9% of patients and was primarily neutropenia. No grade >2 gastrointestinal toxicities or death due to treatment were observed. The most frequent nonhematological adverse event was fatigue. Ten patients responded to treatment with two complete responses (6.3%) and eight partial responses (25.0%), for an overall response rate of 31.3%; 11 patients achieved stable disease (34.3%). The median time to tumor progression and the median survival were 9.2 (95% CI: 6.0-12.4) and 11.8 (95% CI: 7.7-15.9) months, respectively, with a 2-year survival of 22%. On the basis of this trial, the combination of gemcitabine plus irinotecan, administered in a weekly schedule and at this dose, is well tolerated and offers encouraging activity in the treatment of advanced and/or metastatic pancreatic cancer. PMID- 19806787 TI - Weekly administration of docetaxel and epirubicin as first-line treatment for hormone-refractory prostate carcinoma. AB - Androgen-independent prostate carcinoma (AICP) is one of the tumors that continue to respond poorly to chemotherapy. Recently, protocols based on the use of docetaxel have significantly improved survival for patients in this disease. In other types of neoplastic disease, combined therapy with taxanes and anthracycline derivatives has been shown to produce additive effects in terms of growth inhibition, and superior tolerability when associated with weekly administration schedules. These findings prompted us to examine the tolerability and efficacy of weekly treatment of AICP with docetaxel (DOX) plus epirubicin (EPI). We enrolled 35 chemotherapy-naive men with AICP (mean age 72 years, range 68-77) and normal hepatic, renal, and cardiac function. The chemotherapy protocol provided for the IV administration of DOX (30 mg/m2) and EPI (30 mg/m2) on days 1, 8, and 15 every 28 days. Treatment was continued for 6 months or until disease progression and/or unacceptable toxicity was observed. Serum levels of prostate specific antigen (PSA) were monitored in all patients, and reductions from baseline values of >50% were considered indicative of positive responses to treatment. Thirty-four patients were included in the analysis of toxicity, and objective responses to treatment were assessed in the 28 patients with measurable lesions. Nineteen patients (56%) experienced PSA reductions of >50% that persisted for more than 4 weeks. The response to therapy was classified as complete in 1 of the 28 patients (4%) with measurable disease (at the lymph node level). Thirteen others (13/28, 46%) had partial responses, in nine (32%) the disease remained unchanged, and progression was observed in the remaining five (18%); overall response rate was 50% (CR + PR). Of the 27 patients with pain at the time of enrollment, 16 (59%) experienced pain reduction during treatment. The median time to disease progression was 11.7 months (95% CI: 7.7-15.7) while the median survival time was 18.7 months (95% CI: 12.3-25.1). During the study, four patients developed grade 3 anemia and leukopenia, which was reversible in all cases. Lower grades of asthenia, nausea, vomiting, diarrhea, and peripheral edema were also observed. There were no cases of cardiotoxic effects. Alopecia was frequent but reversible in all cases. The results of this preliminary study indicate that the combined administration of DOX and EPI for treatment of AIPC is effective and well tolerated. The weekly administration of the drug combination appears to be a promising approach to the treatment of these tumors. PMID- 19806788 TI - Expression and mutation analysis of TIG1 (tazarotene-induced gene 1) in human gastric cancer. AB - Tazarotene-induced gene 1 (TIG1) has been known to function as a cell adhesion molecule, which leads to better cell to cell contact and reduced proliferation. We investigated expression and mutation status of TIG1 in primary gastric tumors and cell lines to explore the candidacy of the gene as a tumor suppressor. A total of 172 gastric tissue specimes, including 80 primary adenocarcinomas, 12 benign tumors, and 80 adjacent normal mucosa, and 15 gastric cancer cell lines were used. TIG1 expression was analyzed by semiquantitative RT-PCR and immunoblot analysis. To screen for the presence of somatic mutations, RT-PCR-SSCP analysis was carried out. The effect of 5-aza-2'-deoxycytidine treatment was examined to elicit whether TIG1 reduction is associated with abnormal DNA hypermethylation. Compared to noncancerous tissues, a substantial reduction of TIG1 expression was observed in 73.3% (11115) cancer cell lines, and seven of these exhibited nearly undetectable levels of expression. Decreased expression of TIG1 was also found in 62 (77.5%) primary carcinoma tissues compared to adjacent noncancerous tissues, indicating a tumor-specific reduction of TIG1. Expression levels of TIG1 were significantly low in primary carcinomas and cancer cell lines compared to those of normal tissues. Moreover, loss or reduction of TIG1 was significantly high in advanced tumors compared to early tumors and more frequent in poorly differentiated tumors than well or moderately differentiated tumors. TIG1 expression was reactivated or its level was elevated following 5-aza-2' deoxycytidine treatment, indicating that TIG1 expression is transcriptionally silenced in these cancer cells by abnormal DNA hypermethylation. These data indicate that TIG1 undergoes frequent epigenetic inactivation due to aberrant DNA hypermethylation in gastric cancers, and its altered expression is associated with the malignant progression of tumors. PMID- 19806789 TI - A bone metastasis model with osteolytic and osteoblastic properties of human lung cancer ACC-LC-319/bone2 in natural killer cell-depleted severe combined immunodeficient mice. AB - Lung cancer is commonly associated with multiple-organ metastasis, and bone is a frequent metastatic site for lung cancer. Lung cancer frequently develops osteolytic, and less frequently osteoblastic, metastasis to bone. Osteolytic metastasis models of lung cancer have been reported, but no osteoblastic metastasis model is available for lung cancer. In the present study, we established a reproducible model of human lung cancer with both osteolytic and osteoblastic changes in natural killer cell-depleted severe combined immunodeficient mice. Intravenous inoculation of ACC-LC-319/bone2 cells resulted in the development of metastatic colonies in the lung, liver, and bone of the mice. As assessed sequentially by X-ray photographs, osteolytic bone lesions were observed by day 28, and then osteoblastic lesions were detected by day 35. Histological examination revealed the presence of bony spurs, a hallmark of osteoblastic bone metastasis, where osteoclasts were hardly observed. Treatment with an anti-human vascular endothelial growth factor antibody, bevacizumab, as well as zoledronate, inhibited the number of experimental bone metastases, including osteoblastic changes produced by ACC-LC-319/bone2 cells. These results indicate that our bone metastasis model by ACC-LC319/bone2 might be useful to understand the molecular pathogenesis of osteolytic and osteoblastic metastasis, and to identify molecular targets to control bone metastasis of lung cancer. PMID- 19806790 TI - Cloning of a novel splicing variant of RIN1 and its expression in gastric and colon cancer. AB - The regular RIN1 gene is a molecule located on chromosome 1lq13.2, and contains a coding region of 2352 bp with a 3' domain that binds to H-Ras protein, suggesting that it is an important molecule in the intracellular signaling pathway. In this study, we confirmed the existence of a novel form of the RIN1 gene with a different splicing pattern, successfully cloned it, and examined its expression in gastric and colon cancer cell lines. A 612-bp band (the RIN1 variant mRNA) was identified in the RT-PCR product from the colon cancer cell line Colo320D. (A 2352-bp band representing the regular RIN1 gene in HT29 cell line.) The 612-bp band was sequenced and compared with that of the regular RIN1 gene. As a result, the 612-bp product was found to contain a tyrosine phosphorylation site on the 5' side and Ras and 14-3-3 binding domains on the 3' side, indicating that it is a product with a different splicing pattern. The expression of the RIN1 variant mRNA was observed in two of six gastric cancer cell lines and four of five colon cancer cell lines. We identified a novel RIN1 gene with a splicing pattern different from that of the regular RIN1 gene. Comparison of both genes revealed that the novel RIN1 products had a structure conserving the Ras and 14-3-3 binding domains, but lacking two tyrosine phosphorylation sites. Novel RIN1 variant protein was expressed primarily in the cytoplasm and no expression in the cell membrane, and RIN1 variant protein was bound to 14-3-3 protein. In addition, the novel RIN1 mRNA was found to be expressed in gastric and colon cancer cell lines, suggesting that it is an important gene for the function of cancer cells. PMID- 19806791 TI - Comparative proteomic analysis of mouse melanoma cell line B16, a metastatic descendant B16F10, and B16 overexpressing the metastasis-associated tyrosine phosphatase PRL-3. AB - Metastasis is a complex, multistep process by which a cancer cell leaves the primary tumor, travels to a distant site via the circulatory system, and establishes a secondary cancer. A deeper understanding of the molecular events underlying metastasis will provide information that will be useful for the development of new diagnostic and therapeutic strategies. The B16 and B16F10 mouse melanoma cell lines are widely used as model system for studying many aspects of cancer biology including metastasis. Compared with B16, which has a low metastatic potential, the highly metastatic cell line B16F10 displayed a higher metastatic ability along with higher expression levels of the metastasis associated phosphatase of regenerating liver-3 (PRL-3). B16 cells transfected with PRL-3 cDNA (B16-PRL3) had metastatic abilities comparable to those of Bl16F10 cells. To study the molecular mechanisms that underlie metastasis, the proteomes of the B16, B16F10, and B16-PRL3 cell lines were compared using two dimensional differential in-gel electrophoresis. Proteins that varied significantly in levels between these cell lines were selected and identified using mass spectrometry. Interestingly, many proteins, especially those present in membrane fractions, were similarly up- or downregulated in both the Bl16F10 and B16-PRL3 cells lines compared to B16 cell lines. The list of similarly regulated proteins included heat shock protein 70, fascin-1, septin-6, ATP synthase beta subunit, and bone morphogenic protein receptor type IB. These proteins may play a causal role in PRL-3-mediated metastasis. These investigations open an avenue for the further characterization of the molecular mechanisms that underlie metastasis. PMID- 19806792 TI - Knockdown of Bmi-1 impairs growth and invasiveness of human gastric carcinoma cells. AB - The effect of B cell-specific MLV integration site-1 (Bmi-1) RNA interference (RNAi)-mediated inhibition of Bmi-1 expression on the proliferation, apoptosis, and invasiveness of human gastric carcinoma AGS cells was investigated. RT-PCR and Western blot analyses demonstrated that Bmi-1 expression and the Bmi-1 protein level were significantly decreased in Bmi-1 shRNA transfected AGS cells compared to untransfected and nonspecific shRNA transfected AGS cells. Bmi-1 RNAi mediated inhibition of Bmi-1 expression significantly affected cell growth and invasiveness, and resulted in increased AGS cell apoptosis. This was not observed in untransfected and nonspecific shRNA transfected AGS cells. Inhibition of Bmi-1 expression in human gastric carcinoma cells affects cell proliferation and invasiveness. PMID- 19806793 TI - Antitumor effects of targeting hTERT lentivirus-mediated RNA interference against KB cell lines. AB - Squamous cell carcinomas are the leading frequent malignant tumors in the oral and maxillofacial region. Currently available treatment options are of limited efficacy, and there is an urgent need for development of alternative therapies. RNA interference (RNAi) is a sequence-specific RNA degradation process. In this study, we screened and identified an in vitro-transcribed 21-bp shRNA targeting human telomerase reverse transcriptase (hTERT) from three candidates and generated a lentivirus vector. Subsequent experiments indicated that this lentiviral transgenic system could effectively transfer into target KB cells, above 80% gene transfer efficiency at MOI of 2.5, and significantly and specifically inhibited hTERT expression both in mRNA (73.42%) and protein (74.67 82.91%) levels. To further evaluate the role of hTERT-targeted RNAi, we found that hTERT inhibition consequently induced suppression of cyclin D1 (54.67%), upregulation of caspase-3 (100.10%), and caspase-9 (42.67%) of KB cells. Therefore, the apoptosis rates of KB cells were increased by 206.33%. In conclusion, these data indicated the potential of lentivirus vectors in cancer gene therapy, especially after development of more efficient vector production methods, and higher virus titers demonstrated that targeting hTERT RNAi may result in telomere uncapping, which triggers cell cycle arrest and apoptosis signal and leads to tumor suppression. PMID- 19806794 TI - Journey from technician to professional. PMID- 19806795 TI - A conversation with the Richmonds on their 30 years of service with the American Board of Cardiovascular Perfusion. Interview by David A. Palmer and Linda B. Mongero. AB - Beth A. Richmond, PhD, and Mark G. Richmond, EdD, Co-Executive Directors serving for the ABCP, have assumed this role for the past 30 years. Their experience working with a variety of perfusionists in the field influenced the profession and some of the professionals we view as perfusion leaders. Anyone with time working as a clinical perfusionist acknowledges the role they have had establishing the certification process and influencing perfusion education. The goal of this article is simply to highlight the Board's history through the words of both Co-Directors. Our profession, young in years, has a unique history. The names have not been changed and many of the stories have yet to be told. During the winter of 2009, we sat down with Beth and Mark Richmond to talk about their experience working with different Boards over the past 30 years. The following article is their story in their words. PMID- 19806796 TI - Perfusion preservation of the donor heart: basic science to pre-clinical. AB - As a consequence of technology improvements and refinement, perfusion of the donor heart has moved from the research laboratory to clinical studies. Multiple investigators are currently leading pre-clinical trials of devices using perfusion preservation, and one device is now in European clinical trials. One major problem with the donor heart is the high metabolism relative to other organs, and depletion of ATP leads rapidly to acidosis and necrosis of the myocardium. Two techniques in development to address the issue are normothermic and hypothermic perfusion. This review examines the current issues regarding donor heart preservation and techniques of preclinical evaluation necessary for regulatory approval. PMID- 19806797 TI - Clinical efficacy of two-phase leukocyte filtration in high-risk patients undergoing coronary revascularization with cardiopulmonary bypass. AB - The purpose of this study was to investigate the clinical outcome, inflammatory response and myocardial function in high-risk patients undergoing three different leukocyte depletion strategies. Over a four-month period, forty patients (EuroSCORE 6+) undergoing coronary revascularization were prospectively randomized to one of the four perfusion protocols: Group 1 (N=10): Conventional circuits (ECC) + two leukocyte filters (LG6B, Pall, USA) with the method of two phase (continuous + strategic) leukofiltration; Group 2 (N=10): ECC + single leukocyte filter with the method of continuous leukofiltration; Group 3 (N=10): ECC + single leukocyte filter with the method of strategic leukofiltration; Group 4 (N=10) CONTROL: ECC without leukocyte filtration. Blood samples were collected at T1: Baseline, T2: On CPB, T3: X-Clamp, T4: Off CPB, T5: ICU24 and T6: ICU48. Perioperative follow-up was thoroughly monitored. Leukocyte counts in double filter and strategic filtration groups demonstrated significant differences at T4 (p < .05 vs. control). TNFalpha levels were significantly lower in Group 1 at T4 and procalcitonin levels at T5 and T6 (p < .05 vs. control). CKMB levels demonstrated well preserved myocardium in double filter group (p < .05 vs. control). Brain natriuretic peptide levels in double filter group were significantly lower at T5 and T6 with respect to Group 2 (p <.05) and control (p <.001). Matrixmetallopeptidase 9 and D-Dimer levels in double filter group were significantly lower at T5 and T6 (p <.05 vs. control).Two-phase leukofiltration is associated with some compound benefit over continuous deployment in high-risk patients. A larger more powerful study than this pilot one is warranted for further evaluation. PMID- 19806798 TI - Soluble fibrin monomer complex and cardiopulmonary bypass. AB - Soluble fibrin monomer complexes (SFMCs) are precursors of fibrin polymer formation. Laboratory tests can be used to detect SFMCs in plasma. The purpose of this study was to determine whether a positive SFMC test is associated with pre operative, intra-operative, and post-operative variables for patients that have undergone cardiopulmonary bypass (CPB). Pre-operative, operative, post-operative, and laboratory data from 120 consecutive adults patients (July 3, 2006 to June 29, 2007) that had undergone cardiac surgery with the use of CPB were obtained from a prospective quality control database. Two groups were created. Group 1 was all negative (NEG). This group had no SFMC test with a positive result (n=60) and no positive SFMCs (POS, n=60). Group 2 was any positive (POS). This group had at least one positive SFMC test (n=60). The POS group had more patients with endocarditis (11.7% vs. 3.3%, p < .001), chronic obstructive pulmonary disease (COPD) (18% vs. 8.3%, p = .005), longer CPB time (172 +/- 64 vs. 151 +/- 53 minutes, p = .047), and fewer minimally invasive procedures (31.7% vs. 51.7%, p = .002). The POS group required intraoperative (70.0% vs. 53.3%, p = .010) and post operative (75.5% vs. 45.0%, p < .001) transfusions more frequently than the NEG group, despite similar amounts of blood loss. SFMC tests in CPB may be associated with patient pre-operative status and an increase in transfusion requirements. PMID- 19806799 TI - An in vitro study of the effectiveness of carbon dioxide flushing of arterial line filters. AB - Gaseous microemboli (GMEs) have been connected to neurologic impairment and other ischemic complications after surgery. The components of the extracorporeal circuit (ECC) have a large influence on GME production. This in vitro study investigates the use of carbon dioxide flushing of the 38-microm Medtronic Affinity CB351 and 38-microm Medtronic Affinity 351 arterial line filters (ALFs) to decrease GMEs and time for air to clear the ALE An adult circuit was implemented with a silicone oxygenator for vacuum-assisted gas removal and to reduce air before ALE The 48 filters were separated into four equal groups: flushed coated and non-coated and non-flushed coated and non-coated. Carbon dioxide flushing was performed at 6 L/min for 3 minutes. ALFs were retrograde primed at 200 mL/min. An Emboli Detection and Classification Quantifier (EDAC) was used to gather data. The average total emboli and time to clear (seconds) for flush coated were 20.25 +/- 16.78 and 142.17 +/- 174.80 seconds, respectively, flushed non-coated were 30.5 +/- 34.65 and 124.17 +/- 131.40 seconds, non-flushed coated were 162.08 +/- 79.90 and 390.42 +/- 84.36 seconds, and non-flushed non coated were 163.67 +/- 212.67 and 305.92 +/- 179.36 seconds. Flushed filters had an average total emboli count of 25.375 +/- 27.14 and an average time to clear of 13.167 +/- 151.51 seconds. Non-flushed filters had an average total emboli count of 162.875 +/- 157.11 and an average time to clear of 348.167 +/- 143.70 seconds. Coated and non-coated filters for total emboli and time to clear had p values of .86 and .24, respectively. Flushed and non-flushed filters had total emboli and time to clear p values of < .001 and < .001, respectively. No significant difference was found between coated and non-coated filters involving total embolic count and time to clear. A significant difference was found in total embolic count and time to clear between flushed and non-flushed filters. This study shows that fewer emboli and faster embolic clearance time correlate with carbon dioxide flushing of the ALE. PMID- 19806800 TI - Gaseous microemboli in a pediatric bypass circuit with an unprimed venous line: an in vitro study. AB - Miniaturizing cardiopulmonary bypass (CPB) circuits to reduce hemodilution and allogenic blood product administration is common in cardiac surgery. One major concern associated with smaller CPB circuits is a possible increase in gaseous microemboli (GME) sent to the cerebral vasculature, which is exacerbated by vacuum-assisted venous drainage (VAVD). The use of VAVD has increased with smaller venous line diameter and venous cannulae. This study examines the effects of CPB initiation with an unprimed venous line and VAVD in a pediatric circuit. A CPB circuit was set up with reservoir, oxygenator, and arterial filter with a bag reservoir to simulate the patient. All trials were done in vitro, and GME were measured using the EDAC Quantifier by Luna Innovations. EDAC sensors were placed proximal and distal to the oxygenator and distal to the arterial filter. Group 1 was the control group with no VAVD and a primed venous line. Groups 2, 3, and 4 used an unprimed venous line and VAVD of -40, -20, and -10 mmHg, respectively. Total microemboli counts and total embolic load in micrometers were measured at each sensor. Groups 2 (12,379.00 +/- 3180.37) and 3 (8296.67 +/- 2818.76) had significantly more microemboli than group 1 (923.33 +/- 796.08, p < .05) at the pre-oxygenator sensor. Group 2 (57.33 +/- 25.01, p < .05) had significantly more microemboli than group 1 (5.33 +/- 3.21) at the post-oxygenator sensor. No other findings were statistically significant. The results suggest that, if an oxygenator and arterial filter with sufficient air handling capabilities are used, this method to reduce prime volume may not increase GME in the arterial line distal to the arterial filter. PMID- 19806801 TI - A 2007 survey of extracorporeal life support members: personnel and equipment. AB - Extracorporeal membrane oxygenation (ECMO) is used to support patients with cardiopulmonary failure in the intensive care unit. The purpose of this study is to determine what professional qualifications, equipment, and tests are used by established ECMO programs registered with the Extracorporeal Life Support Organization (ELSO). A survey link (Survey-Monkey) was e-mailed to the 110 registered ELSO program coordinators. Forty-nine responses were received. A test of binomial portions showed that nurses were more likely to be ECMO providers than respiratory therapists or perfusionists (p < .05). A chi2 test identified a difference in the type of pump (roller or centrifugal) based on patient age (p < .005). The most common monitoring/safety devices were battery back-up (84%), pre- and post-oxygenator pressure (82%), mixed venous oxygen saturation (80%),venous line pressure (76%), blood flowmeter (63%),bubble detector (61%), point-of-care blood gases (59%), and in-line blood gas monitoring (47%). Laboratory tests available included d-dimer (65%), plasma-free hemoglobin (63%), anti-Xa plasma heparin concentration (43%), thromboelastograph (37%), and heparin concentration using protamine titration (35%). This survey of ELSO-registered centers represents an overview of current ECMO practices. PMID- 19806802 TI - Concomitant repair of mitral valve papillary muscle rupture and tricuspid valve avulsion in a pediatric patient after a motor vehicle collision. AB - We report the case of an 8-year-old patient with severe acute mitral regurgitation and tricuspid regurgitation after a motor vehicle collision caused by traumatic injuries of the supporting structures of both valves. An echocardiogram showed avulsion of the posteromedial papillary muscle of the mitral valve and a severely prolapsed anterior tricuspid valve leaflet. In addition, there was a near-complete defect in the left ventricular side of the interventricular septum, with diffuse hematoma of the akinetic septal muscle. After emergency mitral and tricuspid valve repairs, consisting of re-implantation of the posteromedial papillary muscle and anterior tricuspid leaflet chordal support, the patient completely recovered with no residual valvular pathology. PMID- 19806803 TI - Anesthetic vaporizer mount malfunction resulting in oxygenation failure after initiating cardiopulmonary bypass: specific recommendations for the pre-bypass checklist. AB - Modern technologic advances in medicine have allowed commonly used machines to perform safely with very low risk and a high degree of success. To detect or prevent potential malfunctions, professionals routinely perform pre-use checks for equipment such as anesthesia machines and cardiopulmonary bypass (CPB) machines. These machine checklists are not only critical for a safe operation but also have large impacts on outcomes. For example, when malfunctions are encountered that could have potential negative ramifications or adverse outcomes, multi-approach strategies should be used to identify rectifiable causes and find solutions that are practical. This information can be used to promulgate safe practice guidelines. This case report identifies a machine-based contributing factor to precipitous hypoxia on initiation of bypass in one of our patients. After a detailed approach to identify preventable root causes, we made simple additions to our pre-bypass checklist and recommend these changes to other institutions. PMID- 19806805 TI - Analysis of morphological trait and yield components of mutant wheat under different levels of nitrogen. AB - In order to evaluate the reaction of yield and yield components of wheat's mutant lines to different levels of nitrogen in the crop year of 2006-2007, an experiment was conducted as factorial based on completely randomized block design in 4 replicates in the Research Farm of Agricultural, Medical and Industrial Research School in Zaferaniyeh-Karaj-Iran the first factor included 5 genotypes of wheat (Tabasi-the maternal entity, T-65-7-1, T-65-5-1, T-6-67-60, T-65-58-8) and the second factor consisted of 4 levels of nitrogen (100, 140, 180 and 220 kg ha(-1) urea 46%). The result showed that grain's yield harvest index, number of grain/spike in mutant lines and their parent had significant difference (p < 0.01) in mutant genotypes, meanwhile, the biological yield and the number of spike m2 were not significant difference (p < 0.05). The harvest index, number of grains per spike, grain's yield, biologic yield and number of spikes m(-2), weight of 1000 grains didn't affected by applying different levels of nitrogen fertilizer. PMID- 19806804 TI - Effect of dietary and topical Celecoxib on expression of bcl-2, bax, c-erb-B2 and Ki67 in carcinogen-induced tongue carcinoma in rat. AB - The aim of the study is to determine the effect of Celecoxib administration, dietary or topical, on expression of Ki-67, c-erb-B2, bcl-2 and bax genes in rat tongue by the immunohistochemistry methods and also tdt-mediated dupt-biotin nick end labeling assay in order to explore their role in malignant transformation and the proliferation rate, apoptosis rate in tongue squamous cell carcinoma. Effects of celecoxib on tongue carcinogenesis were investigated in 40 adult male Sprague Dawley 3-3.5 months rats initiated with 30 ppm 4-nitroquinoline-1-oxide. The immunohistochemical expression of Ki-67, bcl2, bax and c-erb-B2 were also examined for analysis of the effects of Celecoxib on tongue carcinogenesis. Differences among groups were statistically analyzed with one-way analysis of variance (SPSS-13, p < 0.05). At week 8, the incidence of tongue precancer lesions was reduced by Celecoxib and there were significant differences in the average expression of Ki-67 (p = 0.00), c-erb-B2 (p = 0.01), bax (p = 0.02), bcl2 (p = 0.02) and also in TUNEL assay (p = 0.00). The results suggest probably that the level of c-erb-B2, bcl-2 and bax expression could show behavior of squamous cell carcinoma in initiation phase of developing carcinoma. PMID- 19806806 TI - The protective effects of garlic extract against acetaminophen-induced oxidative stress and glutathione depletion. AB - Acetaminophen, the most commonly sold over-the-counter antipyretic analgesic, is capable of causing severe and sometimes fatal hepatic damage in humans and experimental animals. The incidence of liver injury due to acetaminophen overdose, either with suicidal intent or by accident, is increasing. Garlic is among those medicinal plants famous for its different health protective effects. In this study, the protective effects of garlic extract on acute acetaminophen induced liver injury were investigated using freshly isolated rat hepatocytes. The hepatocytes were isolated from Sprague-Dawley male rats by a two step collagenase model. Formation of Reactive Oxygen Species (ROS) and Glutathione (GSH) depletion were studied after addition of acetaminophen to cell suspensions. The effects of garlic extract on prevention of ROS formation as well as GSH depletion was investigated and compared with the effects of N-Acetyl Cysteine (NAC) as the standard treatment. Reactive oxygen species formation was assessed by a spectrofluorometry method and garlic extract was shown to be as effective as NAC in decreasing ROS formation induced by acetaminophen. Glutathione (GSH) levels of hepatocytes were determined using HPLC. Garlic extract was effective in preventing GSH depletion significantly (p < 0.05). It is concluded that garlic extract has an antioxidant effect and can protect hepatocytes from GSH depletion following NAPQI production. PMID- 19806807 TI - Comparison of bed planting-furrow irrigation with conventional planting-flood irrigation in durum wheat (T. durum Desf) in southeastern Turkey. AB - There is no clear consensus regarding the advantages of bed planting with furrow irrigation over conventionally irrigated cropping. This 3-year study from Southeastern Turkey aimed to assess the limits to some input savings in bed planting-furrow irrigation in terms of yields and profitability of durum wheat. Field trials were carried out using a randomized complete block design with six treatments and tree replications: T1: Conventional Planting-Flood Irrigation (CP FI) with recommended practices for seed rate, chemical fertilizers and chemical weed control; T2: Bed Planting and Furrow Irrigation (BP-FI) with recommended input rates as in T2; T3: BP-FI with 10% input reduction; T4: BP-FI with 20% input reduction; T5: BP-FI with 30% input reduction; T6: BP-FI with 40% input reduction. The trial had four replications at each location over three cropping seasons, i.e., Akcakale (2004-05, 2005-06) and Koruklu (2006-2007). Individual and combined analysis of variance were performed for grain yields, market prices based on quality assessment, protein content and both 1000-kernel and hectoliter weights. Profitability was assessed with partial budget analysis. Except for yields, there was little effect of treatments on the other variables. Based on yields and economic analysis, the conventional system with flood irrigation was superior to the bed and furrow system, even when the inputs were reduced in such a system. The work demonstrates the site-specific nature of any new technology as there are several local biological and economical factors to be considered. PMID- 19806808 TI - Phytosociological studies of Citrullus colocyanthis L., growing in different altitudinal sites in Saudi Arabia. AB - The aim of the present study was to survey and record the plant species associated with Citrullus colocynthis in different altitudinal localities in the West of Saudi Arabia. Depending on the presence of Citrullus colocynthis L. species, seven stands on the West of Saudi Arabia; expending from 25 m up to 2220 m a.s.l. height and 330 km long were selected for this study. Soil samples were collected from the studied localities and the soil properties were investigated. Also, plant species associated with C. colocynthis were collected, recorded and prepared as herbarium specimens. The studied localities were represented by different ecological, geographical and edaphic sites. A list of 127 species belonging to 41 families present in all locations was recorded. Calotropis procera was the representative species with C. colocynthis in all localities. The percentage of presence of the associated species were different between species in each location and also from location to other. Three different ecological areas could be distinguished in the study area. The area near the red sea characterized by salty sandy soil and low vegetation represented the first area. The second one was the coastal plain and West slope which characterized by low rainfall and xerophytic plants. The third one was the mountainous area which characterized by high altitude, more rainfall and high density of vegetation. PMID- 19806809 TI - Scoliosis curve: before and after surgical correction. AB - The main purpose of this study was to report the surgical experiences and to evaluate the effectiveness of Two-stage operation (including 1-anterior release and fusion with hylofemoral traction and 2-posterior correction and fusion) in scoliosis. The prospective study was performed on 45 scoliotic patients presenting to Tabriz Shohada Orthopedics Teaching Referral Hospital between January 2002 and July 2007. Two-stage operation (anterior release and fusion with hylofemoral traction followed by posterior correction and fusion) was performed for cases in which the scoliosis angle was > 70 degrees and the correction level on bending films was less than 50%. Forty five patients, 32 (71.1%) female and 13 (28.9%) male, with mean age of 15.13 +/- 0.34 years (range of 10 to 18 years), were studied. According to the consecutive sampling, the relation of scoliosis with patients sex was significant (p < 0.005). The mean scoliosis angle was 73.31 +/- 4.21 before operation and 36.11 +/- 2.56 after operation (p < 0.005). The mean preoperative kyphosis angle was 29.95 +/- 3.9 and the mean postoperative kyphosis angle was 24.6 +/- 2.2 (p < 0.005). Scoliosis was significantly more severe in males than females. PMID- 19806810 TI - Effects of reduced calcium and phosphorous diets supplemented with phytase on laying performance of hens. AB - The aim of this experiment was to examine the potential for reduced environmental impact by reducing dietary calcium and phosphorus content and phytase addition of laying hen diets. A randomized complete block design with a 2x2x2 factorial arrangement of 8 dietary treatments: 2 levels ofphytase (0 and 300 FTU kg(-1)) and 2 mineral levels (Ca: 34/18 and NPP: 3.2/2.2 g kg(-1), respectively). A total of 240 White Leghorn (WL) layers, 25 weeks of age were used. Considering birds in 12 cages as a replicate, 5 such replicates were randomly allotted to each dietary treatment. Individual body weight of the bird was recorded at the beginning and end of the experiment. Egg production on an individual basis was recorded daily and percentage hen day egg production was calculated. The cleaned eggshells were dried for 24 h, weighed and expressed as percentage of whole egg. One bird from each experimental unit were selected at random and killed by cervical dislocation at the end of the experiment and the left tibia was removed. Dried bone samples were ashed at 680 degrees C for 12 h for estimation of bone ash. The results of this experiment showed that reducing Ca and NPP (Non-Phytate Phosphorus) without phytase decreased BWG (Body Weight Gain), feed intake, FCE (Feed Conversion Efficiency), egg production, egg shell weight and tibia ash. However, phytase addition to low mineral diets completely corrected the adverse effects associated with low dietary Ca and NPP. It can therefore be concluded that reducing levels of Ca and NPP below current standards and phytase supplementation can reduce pollution potential from laying hen production without adversely affecting bird performance or welfare. PMID- 19806811 TI - Development of morphine induced tolerance and withdrawal symptoms is attenuated by lamotrigine and magnesium sulfate in mice. AB - The goal of this study was to evaluate the effects of lamotrigine and magnesium sulfate on morphine induced tolerance and withdrawal symptoms in mice. Different groups of mice were received morphine (30 mg kg(-1), s.c.) or morphine (30 mg kg( 1), s.c.)+lamotrigine (10, 20, 30 or 40 mg kg(-1), i.p.) or morphine (30 mg kg( 1), s.c.) + magnesium sulfate (20, 40 or 60 mg kg(-1), i.p.) or morphine (30 mg kg(-1), s.c.) + [lamotrigine (10 mg kg(-1), i.p.) + magnesium sulfate (20mg kg( 1), i.p.)] daily for 4 days. Tolerance was assessed using hot plate after administration of a test dose of morphine (9 mg kg(-1), i.p.) on fifth day. Withdrawal zsymptoms (Jumping and Rearing) were assessed by administration of naloxone (5 mg kg(-1), i.p.) 2 h after the last dose of morphine in fourth day. It was found that administration of lamotrigine or magnesium sulfate or their combination decreased the morphine induced tolerance and withdrawal symptoms. From these results it is concluded that lamotrigine and magnesium sulfate alone or in combination could prevent the development of morphine tolerance and withdrawal symptoms. Glutamate release inhibitory effect of lamotrigine and its possible mechanism and property of magnesium, blocking the N-Methyl-D-Aspartate (NMDA) receptor calcium channel, is probably its mechanism on preventing morphine induced tolerance and dependence. PMID- 19806812 TI - Hepatitis C infection and apoptosis in hepatocellular carcinoma. AB - This study was suggested to show apoptotic markers level in Hepatitis C Carcinoma (HCC) patients related to Hepatitis C Virus (HCV) infection and evaluate the Fas and Fas-L levels in each of non-cancerous individuals (controls), HCC and non-HCC patients to show the apoptosis levels which reflects the progression of inflammation and fibrosis in HCV patients. The collected samples were tested by kinetic, ELISA, RT-PCR and other molecular techniques. The obtained results showed that the Fas-upregulations don't change in case of positive viral C infection related to HCC and also in non-HCC. But in negative HCV-RNA, Fas expression in HCC patients is significant. So, the conclusion is neither Fas nor Fas-L may be affected by viral C infection. PMID- 19806813 TI - Oral and intravenous eradication of Helicobacter pylori. AB - The aim of this study was to determine the efficacy of oral regimens in patients without active gastric bleeding and those of intravenous regimens in a group with active gastric bleeding. This study was performed as an interventional investigation in Tehran between April 2004 and March 2008. The patients who had active peptic ulcer were included and divided into two groups. One group included 26 subjects with active gastric bleeding and the other included 29 patients without active gastric bleeding. The intravenous treatment included metronidazole, ampicillin, ranitidine and orally administered bismuth for five days. The oral regimen included metronidazole, amoxicillin, ranitidine and bismuth sub-citrate for 14 days. In intravenous group 24 patients (92.3%) and in oral group 24 subjects (82.8%) had no drug adverse events. The successful treatment was seen in 61.5% (16/26) and 55.2% (16/29) in intravenous and oral methods, respectively. Finally, it may be concluded that both five-day intravenous therapeutic regimen in patients with active gastric bleeding and 14 day oral regimen in patients without active gastric bleeding would have good efficacy and low rate of drug adverse events. PMID- 19806814 TI - Effect of topical aprotinin on early postoperative bleeding and ICU stay after coronary artery bypass graft surgeries. AB - We evaluate the effect of topical application of aprotinin to the heart, pericardia and mediastinum before sternal closure, on early post operative bleeding, blood transfusion requirement and ICU staying time after coronary artery bypass graft surgeries. In a randomized double blinded clinical trial, 128 patients who were scheduled for elective coronary artery bypass graft were allocated into two groups. In group A (aprotinin), patients received 500,000KIU (50 mL) aprotinin and in group S (saline group) the same volume of normal saline was applied. The amount of blood loss collected in chest bottle, the number of pack cells requirement during first 24 h after operation and duration of ICU staying time were recorded. The amount of blood loss in group A (aprotinin) was 451 +/- 218 mL compared with 707 +/- 269 mL in group S (saline) (p = 0.003). The number of pack cells consumption was 0.5 +/- 0.7 units in group A (aprotinin) compared with 1.7 +/- 1 units in saline group (p = 0.002). Intensive Care Unit (ICU) staying time was 48.8 +/- 13.6 h in group A (aprotinin) and 69.4 +/- 16.6 h in saline group (p = 0.001). This study showed that topical application of aprotinin at the end of coronary artery surgeries, significantly reduce postoperative bleeding and blood transfusion requirement during first 24 h after operation and also ICU staying time. PMID- 19806816 TI - I was a teenage death panelist. PMID- 19806815 TI - Feeding value and in vitro digestibility of date-palm leaves supplemented with different supplementary energy. AB - The aim of this study was to evaluate the feeding value and also the effect of different supplementary energy on increasing in vitro DM digestibility of date palms leaves obtained in pruning. A randomized complete design with different energy supplementary with 3 replicates was conducted. Harvested date-palm leaves were air-dried, ground through a 5 mm screen, then palm leaves (L) ensiled with different levels of urea (U) and other supplementary energy. Combination, mixture was packed tightly in 21 (10 L) plastic containers. The containers were opened after a fermentation of 60 days. Treatments consisting: L (Palm leaves)+5% U (urea) (T1), L+5% U+5% BP (Beet pulp) (T2), L+5% U+10% BP (T3), L+5% U+5% M (Molasses) (T4), L+5% U+10% M (T5), L+5% U+5% CP (Citrus pulp) (T6) and L+%5 U+10% CP (T7). The results of this experiment showed that supplementation of date palm leaves with energy supplements significantly (p < 0.05) increased IVDMD and T5 had higher digestibility than those of the other treatments. ADF content of silage was not affected by dietary treatments. However, NDF content was significantly (p < 0.05) affected by treatments and T2 and T6 had higher NDF content compared with the other treatments. Supplementation of palm leaves with energy supplementary had no significant effect (p > 0.05) on Crude Protein (CP) content of silages. However, these supplements had a significant effect (p < 0.05) on calcium and phosphorus levels of silage materials and T7 had higher Ca and P content compared with the other treatments. In conclusion, supplementation of date-palm leaves with energy supplements can improve IVDMD and feeding value of this feed source. PMID- 19806817 TI - The case for killing granny. Rethinking end-of-life care. PMID- 19806818 TI - No country for sick men. To judge the content of a nation's character, look no further than its health-care system. PMID- 19806819 TI - Born to be big. Early exposure to common chemicals may be programming kids to be fat. PMID- 19806820 TI - Vitamin A supplementation programme--policy, challenges and concerns. PMID- 19806821 TI - Correlates of inpatient healthcare seeking behavior in India. AB - OBJECTIVES: The present study was attempted to study the in-patient health care behaviour, particularly choice of health care providers and their determinants. METHODS: An analysis for treatment seeking behavior of 5989 inpatients which implied to 6726 hospitalized cases due to five ailments namely, heart, diarrhea, tuberculosis, urinary and gynecology diseases in India during the last 365 days prior to survey date has been revealed in this paper using data from the 60th round of National Sample Survey, 2004. Multivariate logistic regression model has been adopted to understand the correlates of being inpatients and of opting private hospitals. RESULTS: It is evident that crowded household, being female, having higher educational level, living in lower income households, and living in urban residents have lower chances of being hospitalized compared to their respective counterparts. Maximum people opted for expensive private health sector which accounts for around 59 per cent. This study also reveals that many people had some form of treatment before being an inpatient where 59 per cent had past treatment. CONCLUSIONS: Patients from the highest quintile of MPCE have higher chances of being hospitalized and also of choosing costly private health care sector. Significant differences were found within age, sex, education, residence and monthly per capita consumption expenditure for hospitalization. Older age groups, highly educated and urban patients have higher instances of choosing a private sector. PMID- 19806822 TI - Determination of sex-ratio by birth order in an urban community in Manipur. AB - OBJECTIVE: To determine the sex ratio by birth order and to assess the sex preference of the couples in an urban community. METHODS: A cross sectional study, in an urban community in Manipur, was conducted among the currently married couples. Data on background characteristics of the couple, family pedigree chart (of the offspring) including history of abortion, stillbirth, death of child of the couple, sex preference and Pre-natal Diagnostic Techniques (Regulation and Prevention of Misuse) Act [PNDT Act] were collected through a structured interview. Data were analyzed using descriptive and chi-square statistics. RESULTS: There were a total of 1777 births to the 855 couples interviewed. There were 900 females per 1000 males for the 1st birth order but the sex ratio was favorable towards females in the 2nd, 3rd and 4th birth orders. Among both the husbands and wives, being more educated was significantly associated (p<0.05) with preferring lesser number of children, using new technology for sex selection and having heard of the PNDT Act. Majority of those who wanted to use new technology for sex selection (128, 56.6%) preferred to have male child. CONCLUSION: Sex ratio in this community was favorable towards females, though it was less among the first born babies. PMID- 19806823 TI - Prevalence of psychosocial problems among adolescents in district Dehradun, Uttarakhand. AB - OBJECTIVE: To find out the prevalence of psychosocial problems among adolescents. METHODS: A cross sectional study was conducted at Doiwala block, Dehradun during 2006-2007 among 840 adolescents, selected by multistage random sampling. Data was collected on a structured and pre-tested questionnaire by interviewing the adolescents and conducting their thorough clinical examination. Epi Info statistical software package version 3.4.3 was used for analysis. RESULTS: The overall prevalence of psychosocial problems among the adolescents was found to be 31.2%. The psychosocial problems were more in males (34.77%) as compared to females (27.6 %). CONCLUSIONS: There is need of strengthening the existing "package" of services for adolescents in various initiatives and programmes. PMID- 19806824 TI - Universalizing access to primary health care in India. AB - Life expectancy is improving steadily but India needs rapid gains in health. Communicable diseases, maternal, perinatal and nutritional deficiencies continue to be important causes of deaths; non-communicable diseases and injuries are also taking a substantial toll of human lives. A basic health care package based on a core set of cost-effective health interventions can address existing disease burden. Its implementation requires increased investment and enhanced health care delivery capacity. Large diversity in India calls for local adaptation of the basic health care package and its delivery mechanism. To encourage accountability, access should be monitored at district level by an independent agency. PMID- 19806825 TI - Profile of behavioural risk factors of non-communicable diseases in an urban setting in New Delhi. AB - The present study was conducted to assess the prevalence of behavioural risk factors of non communicable diseases among urban adult population. The study participants included 531 adults residing in Mata Sundari road, which is an urban colony in New Delhi. They were interviewed using the WHO STEPS 1 questionnaire on lifestyle factors. Smoking prevalence was found to be 18.4% out of which over three fourth smoked more than one packet of cigarettes per day. Almost one third of known hypertensive patients were not on any treatment regimen. 80.6% did not undertake any kind of physical activity. 43% consumed only one serving of green vegetable in a day while 58% included fruits as a part of diet only once or twice in a week. There is a need to develop strong community based lifestyle behavioural intervention programs. PMID- 19806826 TI - Death certification in a teaching hospital--a one year review. AB - Death certification is believed to be very poor and inaccurate in most of the health institutions in India. A hospital based study on assessing the accuracy in completing the medical certificates of cause of death was conducted in Regional Institute of Medical Sciences (RIMS), Imphal during September to December 2007. Medical Certification of Cause of Death (MCCD) from the Medical Record Section for the period from 1st January up to 31st December, 2006 were audited for errors committed in completing the certificates. Data extracted using a proforma--age, sex, date of death, causes of death as recorded, department. Major error was observed in 38.3% and minor error was observed in 77.6% of the MCCD. Mechanism of death was listed as the cause of death in 21.9% of the certificates. The most prevalent type of error was the absence of time intervals (65.3%). PMID- 19806827 TI - Iodine status of women in reproductive age group in urban slums of Cuttack City, Orissa. AB - A community based cross-sectional study involving 168 women of reproductive age was carried out in urban slums of Cuttack city in 2005 to assess the iodine status. Pre-designed, pre-tested schedule was used to collect relevant information & urine samples collected from the study subjects were analyzed by ammonium persulfate digestion method to estimate the iodine level. The median urinary iodine concentration of the study subjects was 64.5 microg/L, 62.5% of study population had iodine deficiency i.e. < 100 microg/L & among them, 74.3% had moderate to severe iodine deficiency i.e. <50 microg/L. Iodine deficiency was significantly higher among women of Muslim religion & Hindu Scheduled Caste & those coming from joint families. PMID- 19806828 TI - Prevalence of hypertension and correlates among adults of 45-60 years in a rural area of Tamil Nadu. AB - Hypertension is one of the major causes of cardiovascular morbidity and mortality. Community based studies in the rural areas of Tamil Nadu on the prevalence of hypertension and its associated risk factors are scarce. A cross sectional study was undertaken among a sample of 406 individuals (45-60 years) selected by the standard 30 cluster systematic random sampling technique to find out prevalence of hypertension and its associated risk factors in a rural area of Tamil Nadu. Chi-square test and multiple logistic regression were employed using SPSS package. The overall prevalence of hypertension was 33% and higher among sedentary type (41%). In bivariate analysis many of the independent variables correlated with hypertension, but in multivariate analysis, only body-mass index, family history and age remained significant. PMID- 19806829 TI - Health metrics improve childhood immunisation coverage in a rural population of Andhra Pradesh. AB - Infant and child mortality, including deaths due to vaccine-preventable diseases, remains high in the rural areas of India. In Andhra Pradesh, the immunisation coverage of children in the 12-23 month age-group has progressively declined to 43%, indicating the immediate need for taking corrective steps. The Rural Effective Affordable Comprehensive Healthcare (REACH) project, based on the health metrics strategy was initiated to improve childhood immunisation coverage. Information on the immunisation status of children was recorded and each child was tracked with the help of a computerised database to provide timely immunisation. The health metrics strategy resulted in complete immunisation (96%) of all the 698 children in the 12-23 month age-group residing in the villages of the Medchal Mandal in the year 2007, as compared to 43% reported by the National Family Health Survey-3. Action oriented health metrics coupled with information technology can thereby improve childhood immunisation significantly. PMID- 19806830 TI - Socio-demographic profile of 15-24 years old male narcotic substance users in a resettlement colony of Delhi. AB - A community based cross-sectional study was conducted during April 2004-March 2005 in an urban resettlement colony in East Delhi to study the narcotic substance use and the socio-demographic characteristics of users. 208 male narcotic substance users aged 15-24 years were studied. 59.1% of the narcotic substance users were between 21-24 years of age. 78.8% were using ganja and 39.9% each heroin & bhang. 37% were abusers; 36% were dependent users. 58.7% were single at the time of interview. 48% were either illiterate or just literate; 64% belonged to middle socio-economic status. 68% had initiated narcotic substance use out of curiosity. PMID- 19806831 TI - Trend and causes of maternal mortality among women delivering in S. N. Medical College Hospital, Agra. AB - A retrospective data analysis from records of patients from medical record section of department of gynecology and obstetric, S. N. Medical College and Hospital, Agra was done to find out the trend and causes of maternal mortality occurred during 1999-2007. The maternal deaths in the context of different causes were analyzed. A total of 192 maternal deaths occurred on 6386 live-births during last 9 years which gives anoverall hospitalized Maternal Mortality Ratio (MMR) as 30.07 per 1000 live births during the period. Out of these total deaths more than half (51.04%) were due to indirect causes. Anaemia (47, 24.48%), hemorrhage (35,18.23%), toxemia (35,18.23%), septicemia (18, 19.23%) were the main causes. PMID- 19806832 TI - Satisfaction of in-patients concerning patient care in a tertiary care hospital in Punjab. AB - Patient satisfaction is defined as a patient's personal evaluation of health care services and providers and thus a vital component of health care outcome and remains an area of interest and need in health care. Hence to assess in-patient satisfaction a study was conducted in a private tertiary care hospital on a stratified random sample of 100 in-patients drawn from four different general wards using a structured questionnaire. Overall the patients were found to be generally satisfied with care and services provided with a score of 87.12%. PMID- 19806833 TI - Prevalence of coronary risk factors in a rural community of Andhra Pradesh. AB - A community based cross sectional survey was carried out to study the prevalence of the risk factors for CHD and identify the high-risk groups in the rural community. 924 subjects of 30 years and above were investigated using structured, pre-tested schedule. Coronary risk factors were assessed. Statistical analysis was done by using Chi square test for prevalence of coronary risk factors in the age and sex groups. The prevalence of coronary risk factors ranged from 5.4 to 41.3%. Tobacco use, alcohol consumption, truncal obesity and sedentary life style were significantly prevalent among males. Whereas truncal obesity and hypertension among females. Behaviour change communication strategies targeting these high risk factors need to be emphasized to lower CHD related morbidity burden in the community. PMID- 19806834 TI - An investigation into a mass psychogenic illness at Burdwan, West Bengal. AB - This investigation of the outbreak of mass psychogenic illness during July 2007 at three schools in Burdwan was done to observe the description of the epidemic, identify its characteristics and to study any particular factor associated with the spread of illness. Relevant data were obtained from medical and school records. All concurrent factors were considered. All the incidents happened prior to the examinations. There have been differences in presentations and final diagnosis among the schools. Intense media coverage influenced the manifestations. Poor school performance was associated with recurrence of symptoms. So understanding the academic anxiety is important to prevent such outbreaks. PMID- 19806835 TI - Characteristics of malaria cases attending OPD of a tertiary level hospital in tribal area of Jharkhand. PMID- 19806836 TI - How pot became legal. PMID- 19806837 TI - Malpractice reform. PMID- 19806838 TI - Who's afraid of the flu? PMID- 19806839 TI - The vaccination war. PMID- 19806840 TI - Pipeline programs in the health professions, part 1: preserving diversity and reducing health disparities. AB - BACKGROUND: Racial and ethnic minorities are underrepresented in the health professions. Affirmative action and educational pipeline programs play a vital role in increasing the diversity of health professions, addressing educational opportunity gaps, and reducing health disparities. Part 1 of this 2-part series discusses the need for educational pipeline programs to assist underrepresented minorities (URMs) in entering the health professions and the importance of these programs in developing a cadre of diverse providers to reduce health care inequality. METHODS: Part 1 presents an overview of diversity in the medical and health care workforce, educational enrichment programs, key components of successful pipeline programs, and notable pipeline examples for underrepresented students at the University of Nebraska Medical Center. Recommendations for improving and developing pipeline programs are also included. Part 2 reviews affirmative action case law and legislation along with recommendations for maintaining and reviewing diversity pipeline programs in light of recent anti affirmative action challenges. CONCLUSION: Pipeline programs are an important strategy for addressing the shortage of URMs in the health professions. Anti affirmative action initiatives threaten the existence of these student preparation programs and the ability of our nation to produce physicians of color and other health care providers who are more likely to serve in underrepresented communities and work to reduce related health disparities. Programs at universities and academic medical centers must develop innovative partnerships with underserved communities, adopt strategies that demonstrate a strong commitment to increasing racial and ethnic minorities in the health professions, and develop viable funding mechanisms to support diversity enrichment programs. PMID- 19806841 TI - Pipeline programs in the health professions, part 2: the impact of recent legal challenges to affirmative action. AB - BACKGROUND: Despite recent challenges to educational pipeline programs, these academic enrichment programs are still an integral component in diversifying the health professions and reducing health disparities. This is part 2 of a 2-part series on the role of pipeline programs in increasing the number of racial and ethnic minorities in the health professions and addressing related health disparities. Part 1 of this series looked at the role of pipeline programs in achieving a diverse health professional workforce and provided strategies to expand pipeline programs. METHODS: This paper presents an historical overview of affirmative action case law, anti-affirmative action legislation, and race conscious and race-neutral admission programs in education. Additionally, part 2 reviews current legal theory and related law that impact the diversity and cultural competence pipeline programming at higher-education institutions. Finally, based on recommendations from a review of legal and other literature, the authors offer recommendations for reviewing and preserving diverse pipeline programs for health professional schools. CONCLUSION: Affirmative action is an essential legal means to ensure the diversity-related educational programs in the health profession educational programs. Anti-affirmative action legislation and state-sponsored antiaffirmative voter initiatives have the potential to limit the number of underrepresented minorities in the health professions and create even greater opportunity gaps and educational disparities. Therefore, we must shift the paradigm and reframe the dialogue involving affirmative action and move from debate to a collaborative discussion in order to address the historical and contemporary disparities that make affirmative action necessary today. PMID- 19806843 TI - The brief CV review session: one component of a mosaic of mentorship for women in academic medicine. AB - BACKGROUND: In the current climate of increasing demands on a disproportionately small number of senior female faculty, we implemented a brief curriculum vitae (CV) review session as an opportunity to expand the professional network of junior women faculty and provide them with additional formal career advice. METHODS: For 3 years, junior (mentees) and senior (mentors) faculty from different departments were paired in half-hour CV review sessions, as part of an annual conference focused on professional development for faculty women. Participating faculty received questionnaires to assess their experience with the sessions, and their feedback was combined over all 3 years and compared using chi2 and Fisher's tests. RESULTS: During the 3 years, there were 93 CV review sessions. Although 84% of the mentees reported having a mentor, only 62% of mentees reported that any previous mentoring experience was helpful. Most (90%) participated in the CV review to determine if their career was "on track." The mentees reported that the CV review session was helpful (93%), provided new information (87%), and identified that they were "on track" for promotion (75%). The mentors felt that their mentees were progressing appropriately in their career (78%) and provided specific recommendations for the mentees (100%). The majority (78%) of mentors felt comfortable mentoring junior faculty outside their department. CONCLUSIONS: Brief interventions, such as a CV review session, can provide additional counsel to junior faculty, helping them assess their career progress as part of a mosaic of mentorship. PMID- 19806842 TI - Historically black medical schools: addressing the minority health professional pipeline and the public mission of care for vulnerable populations. AB - Substantial changes in not only access to care, cost, and quality of care, but also health professions education are needed to ensure effective national healthcare reform. Since the actionable determinants of health such as personal beliefs and behaviors, socioeconomic factors, and the environment disproportionately affect the poor (and often racial/ethnic minorities), many have suggested that focusing efforts on this population will both directly and indirectly improve the overall health of the nation. Key to the success of such strategies are the ongoing efforts by historically black medical schools (HBMSs) as well as other minority serving medical and health professional schools, who produce a disproportionate percentage of the high-quality and diverse health professionals that are dedicated to maintaining the health of an increasingly diverse nation. Despite their public mission, HBMSs receive limited public support threatening their ability to not only meet the increasing minority health workforce needs but to even sustain their existing contributions. Substantial changes in health education policy and funding are needed to ensure HBMSs as well as other minority-serving medical and health professional schools can continue to produce the diverse, high-quality health professional workforce necessary to maintain the health of an increasingly diverse nation. We explore several model initiatives including focused partnerships with legislative and business leaders that are urgently needed to ensure the ability of HBMSs to maintain their legacy of providing compassionate, quality care to the communities in greatest need. PMID- 19806844 TI - Increasing the capacity of health sciences to address health disparities. AB - In order to create a cohort of investigators who are engaged in health disparities research, scholarship, and practice, and to increase the amount of funding in the university that is invested in research focused on reducing health disparities, the San Diego EXPORT Center implemented 2 major initiatives: (1) the support of underrepresented minority (URM) junior faculty development and (2) the funding for pilot research grants in health disparities. This paper describes the activities employed by the center and summarizes the outcomes of these two initiatives. Ninety-five percent (18 of 19) URM junior faculty completed the faculty development program, and 83.3% (15 of 18) of the completers are advancing in their academic careers at University of California San Diego (UCSD) and are teaching, working with populations at risk and/or conducting research in health disparities. EXPORT awarded 7 investigators a total of $429186 to conduct pilot research, and 71.4% (5/7) have now obtained $4.7 million in independent extramural funding. The San Diego EXPORT Center has increased the research capacity, strengthened the infrastructure for health disparities research, and created a cohort of successful URM junior faculty who are advancing in their academic careers. These investigators are already changing the climate at UCSD by their leadership activities, research focus, peer-networking, and mentoring of students. PMID- 19806845 TI - Cultural competency in health care: evaluating the outcomes of a cultural competency training among health care professionals. AB - PURPOSE: The purpose of this research was to examine the effectiveness of a cultural competency training program designed to improve the knowledge and skills of health care providers and administrators engaging in cross-cultural, clinical encounters. METHODS: We conducted an evaluation study among 43 health care professionals (health care providers and health administrators) who attended a 4 hour cultural competency workshop. We used a post-then-pre method of self reported evaluation to answer a key question: Does cultural competency training produce a measurable change in knowledge and skills relating to the care of patients from diverse cultural and ethnic backgrounds? RESULTS: The study findings suggest that there are statistically significant change in participants' self-report of knowledge and skills related to cultural competency. CONCLUSIONS: Our study shows that a cultural competency training program that integrates key topics as recommended by the Institute of Medicine and includes Culturally and Linguistically Appropriate Services in Health Care standards improves the knowledge and skills of health care providers and administrators. Following the training, the participants self-reported not only an enhanced understanding of the health care experiences of patients with diverse backgrounds, but also an improvement in their skills to effectively work in cross-cultural situations. In addition, our study offers a reliable, innovative, and time-efficient strategy- post-then-pre method-for evaluating the outcomes of 4-hour cultural competency training programs. PMID- 19806846 TI - Physicians' cultural competency as perceived by African American patients. AB - The purpose of the study was to determine the association between African American patients' perceptions of physician cultural competency and patient satisfaction with the visit, independent of other factors, including physician and patient race concordance. African American participants were surveyed at urban clinics. Cultural competency (Perceived Cultural Competency scale) was based on the 3-factor model that includes patients' perception of (1) physicians' cultural knowledge, (2) physicians' cultural awareness, and (3) physicians' cultural skill. The results confirmed that patients' perceptions of physician cultural competency are independently associated with satisfaction with the visit. These results further validate use of the Perceived Cultural Competency scale as a tool to measure patients' perceptions of physicians' cultural competency. PMID- 19806847 TI - Perceptions of pediatric clinical research among African American and Caucasian parents. AB - Difficulty in recruiting African American adults for clinical trials is well documented, but there is no consensus on African American children. Responses of a survey completed by 90 African American and Caucasian parents from December 2004 to April 2005 were analyzed to determine if racial disparities exist in research participation interest in an academic pediatric dermatology clinic. The majority of questions (32 of 38) were answered similarly by subjects of both races. However, when compared to African Americans, Caucasians were slightly more trusting (84% vs 65%) in regard to either total or moderate trust (p = .03). African Americans were 3 times as likely to feel that their child might be "treated like a guinea pig" if the child was a research subject (p = .03). Nearly a third more Caucasians than African Americans would be more inclined to enroll their healthy child in a research study if they had an established relationship with the health care provider informing them of the study (p = .0001). Caucasians had more exposure to research (p = .03). Nevertheless, there was no racial difference in the willingness to theoretically allow their child to participate in research studies. Accordingly, the possible lack of trust should not be used as the only reason racially representative recruitment goals are not accomplished. PMID- 19806848 TI - Addressing the challenges of Latino health research: participatory approaches in an emergent urban community. AB - BACKGROUND: Challenges to recruitment of Latinos in health research may include language, cultural and communication barriers, trust issues, heterogeneity of legal status, and a high percent of uninsured when compared to the US population. This paper highlights the community-based participatory research (CBPR) process and expands on the applicability of these principles to Latino communities. METHODS: We review steps taken and describe lessons learned in using a participatory approach to broadly assess and address the health of urban-dwelling Latinos in Baltimore, Maryland, through the adaptation of CBPR principles. FINDINGS: We identified health priorities, access barriers, and community resources (eg, Latin American trained nurses who were not currently working in the health field, immigrant networks) using a participatory approach. Suggestions for improving trust, research participation, and access to care ranged from not collecting data on legal status, and regular attendance and presentations of ongoing research at community provider meetings, to referral to free or low-cost health care services at screening events. CONCLUSION: Despite growing interest in CBPR, limited guidance exists on how to apply CBPR principles to conduct health research among Latinos. Incorporating a participatory process can help address Latino community concerns, enrich quality and relevance of research, and empower community members. PMID- 19806849 TI - Racial/ethnic differences in parental concern about their child's drug use in a nationally representative sample in the United States. AB - INTRODUCTION: Parental concern and negative attitudes toward drug use may prevent youth from being involved in drug use. However, few studies have addressed parental concern about children's drug use and its possible variation by race/ethnicity. In this study, we explored the potential racial/ethnic differences in parental concern about their children's drug use with a nationally representative sample. METHODS: The data were from the 2003 National Survey of Children's Health, a random household telephone survey of parents of children up to age 17 (n = 102353). The analytic sample was restricted to parents of children aged 6 to 17 years (n = 61046). Multivariate logistic regression models, controlling for children's age, gender, family structure, and family poverty level, were fitted, simultaneously accommodating the complex survey design. RESULTS: Parents of African American and Hispanic children expressed more concern than parents of white children, even after controlling for potential confounders (adjusted odds ratio (AOR), 1.9; 95% CI, 1.8-2.1 and AOR, 1.9; 95% Cl, 1.7-2.1, respectively). CONCLUSIONS: The level of parental concern about adolescent drug use was different across race/ethnicity groups. The results may have implications for parental participation in school-based adolescent prevention programs. PMID- 19806850 TI - Review of the literature on cultural competence and end-of-life treatment decisions: the role of the hospitalist. AB - OBJECTIVE: To determine whether any associations exist between cultural (racial/ethnic, spiritual/religious) competence and end-of-life treatment decisions in hospitalized patients and the potential impact of those associations on hospitalists' provision of care. DATA SOURCES AND METHODS: MEDLINE, PubMed, Embase, Psychlnfo, and CINAHL databases were searched using the following search terms: cultural competence, race, ethnicity, minority, African American, Hispanic, end of life, palliative care, advanced care planning, inpatient, religion, spirituality, faith, hospitalist, and hospice. We identified studies in which spirituality/religion or race/ethnicity was used as a variable to study their potential impact on end-of-life treatment decisions in hospitalized patients. RESULTS: In only 13 studies was spirituality/religion or race/ ethnicity used to study its effect on end-of-life decisions in hospitalized patients. African American patients tended to prefer the use of life-sustaining treatments at the end of life, and race/ethnicity did not appear to affect decisions to withhold or withdraw certain types of life-sustaining technology. Specific spiritual needs were identified both within and outside organized religions when members of those religions were hospitalized at the end of life. CONCLUSIONS: End-of-life care may present unique challenges and opportunities in culturally discordant hospitalist-patient relationships. Culturally competent health care in an increasingly diverse population requires awareness of the importance of culture, particularly spirituality/religion and race/ethnicity, in the care of hospitalized patients at the end of life. PMID- 19806851 TI - Culturally specific interventions for African American smokers: an efficacy experiment. AB - This pilot study sought to dismantle the efficacy of culturally specific print materials for smoking cessation. Two-hundred sixty-one African American smokers were randomized into 1 of 2 conditions: standard booklet or culturally specific booklet. The content and length of the interventions were identical yet varied in their degree of cultural specificity. Three-month follow-up assessments were completed by 70% (N = 183) of participants. Dependent variables included content evaluation, readiness to quit smoking, and actual behavior change. Evidence suggested that the culturally specific material was more effective at capturing attention, providing encouragement and gaining interest compared to standard materials; however, greater credibility was found for standard materials. In addition, greater readiness to quit and more 24-hour quit attempts were found in the standard condition. No differences were found in abstinence rates. In conclusion, culturally specific interventions may be preferred over standard approaches among African American smokers. Culturally specific approaches, however, may not result in greater behavior change. Implications for written interventions and cultural specificity are discussed. PMID- 19806852 TI - Prostate cancer treatment patterns among racial/ethnic groups in Florida. AB - BACKGROUND: Prostate cancer is the second leading cause of cancer death among men in the United States. Blacks have the highest incidence and mortality rates. Treatment differences have been observed between black and white men. Brachy monotherapy (BMT) has become popular for localized prostate cancer because of its convenience, being the least invasive, and resulting in better quality of life during and after treatment. No studies have specifically examined BMT in treating localized prostate cancer by race/ethnicity. OBJECTIVES: We sought to (1) describe treatment patterns among men with localized prostate cancer, (2) identify factors affecting the use of BMT, and (3) examine if there was any difference in BMT use by race and ethnicity. METHODS: Florida cancer incidence data of 1994-2003 were used to extract information on men diagnosed with localized prostate cancer along with their demographics, primary payer at diagnosis, tumor stage and treatments. Logistic regression was performed to assess the likelihood of receiving BMT. RESULTS: The study found that surgery and radiation were the 2 major single treatments for localized prostate cancer. The percent of patients receiving BMT treatment increased from 1994 through 2003. Men with the following characteristics were more likely to receive BMT than their counterparts: Non-Hispanic white, older, married, Medicare beneficiaries and military personnel, with well-differentiated tumor, and receiving treatment in facilities with high practice volume and/or located in urban counties. CONCLUSION: There were racial/ethnic differences in localized prostate cancer treatment. Possible reasons for the differences require further research. PMID- 19806853 TI - Predictors, barriers, and facilitators of lipid-lowering medication use among African Americans in a primary care clinic. AB - INTRODUCTION: Hypercholesterolemic African Americans are less likely than white Americans to be taking lipid-lowering medications, yet they suffer disproportionately from coronary heart disease (CHD). METHODS: Through medical record abstraction and focus groups with patients and physicians, we sought a better understanding of the predictors, barriers, and facilitators to lipid lowering medication use in a Detroit primary care clinic. Stepwise regression analysis included 634 African American patients with abnormal cholesterol values (n = 575) or currently prescribed a lipid-lowering medication (n = 59). Focus group transcripts were analyzed with a framework approach. RESULTS: Overall 174 (30.3%) of hypercholesterolemic African Americans were prescribed a lipid lowering medication. Patients with hypertension or CHD were significantly more likely to have a lipid-lowering medication prescription than those without, adjusted prevalence ratio 2.56 (95% CI, 1.76-3.74) for hypertension and 1.70 (95% CI, 1.45-2.00) for CHD. Focus groups revealed 2 barriers to lipid-lowering medication use named by both physicians (n = 12) and patients (n = 23): cost and forgetting to take medication, often because of lack of symptoms. CONCLUSIONS: Physicians and patients suggested better education by physicians and at the community level to improve lipid-lowering medication use. Simple and direct patient-physician discussions emphasizing long-term benefits are recommended. PMID- 19806854 TI - Avoidance of invasive diagnostic and harmful therapeutic measures in a patient with malignant fibrous histiocytoma and extensive clinically occult metastases. AB - Malignant fibrous histiocytomas are rare soft-tissue sarcomas of uncertain histogenesis but represent the most common soft-tissue sarcomas in adults. In this report of a 70-year-old male patient suffering from this rare malignant disease, we describe the diagnostic benefits of combined functional and morphological imaging as realized in positron emission tomography and computed tomography, helping to avoid invasive diagnostic methods and, most importantly, harmful and quality-of-life-reducing, oncologically ineffective therapies. PMID- 19806855 TI - Venous thrombosis and pulmonary embolism in a child with pneumonia due to Mycoplasma pneumoniae. AB - A 13-year-old boy with Mycoplasma pneumoniae pulmonary infection developed deep vein thrombosis and pulmonary embolism. He was found to have protein S deficiency and transient antiphospholipid antibodies. Though uncommon, it is important to consider venous thromboembolic disease in children whose clinical course is atypically severe. PMID- 19806856 TI - Rocking the match II: more lessons on applying and getting into residency. AB - Everyone can think of someone who was just plain smart and who did really well in school without even studying. On the other hand, we all know that most good students work at it, even if they are very bright. In the same vein, there are some students who don't have to prepare for the application process. They may be good applicants for residency training based solely on their academic record, the support they will receive from faculty, and on interpersonal skills that will allow them to shine during an interview. However, there aren't a lot of people like that. The major message to medical students reading this article is that becoming a good applicant for residency training, and indeed for other things in life, is a learnable skill. The responses of the medical students to the 10 trigger questions in this article demonstrate that. We all hope that the tips we provide here will help many students achieve the level of success they deserve. PMID- 19806857 TI - Health/health care disparities: are predominantly African American institutions making a difference? PMID- 19806858 TI - The big keep getting bigger. IPO viewed as positive in long-term-care arena. PMID- 19806859 TI - Bottom line blues. Crisis took big bite out of not-for-profit portfolios. PMID- 19806860 TI - Triple play by the bay. Parties debate San Leandro's future, ER closure. PMID- 19806861 TI - Just like home. Long-term-care facilities try new approaches in their operations to shed that institutional image. PMID- 19806862 TI - The fix isn't in. Likelihood for doc-payment solution remains low. PMID- 19806863 TI - These are volatile times and uncertainty is its hallmark. Tolerating it is crucial. PMID- 19806864 TI - Prescription opioids and physician responsibility. PMID- 19806865 TI - Chronic kidney disease: the new epidemic and its impact on West Virginia. AB - The prevalence and incidence of chronic kidney disease (CKD) is growing at an alarming rate. Estimates suggest that CKD affects an estimated 13 percent of Americans, and West Virginia leads the way, with the highest per capita rate in the country of patients with kidney failure starting dialysis. There is a great lack of awareness about the risks of CKD among the general population, many of whom are unaware of their risk status or even the presence of CKD. The increasingly older, diabetic and obese populations likely account for the high prevalence of advanced CKD in West Virginia, as well as the fact that a large percentage of the state's population lives 2-3 hours' distance from specialized care. Additionally, there are relatively few physicians in West Virginia specifically trained to treat the growing numbers of patients with kidney disease, which is usually silent until well past the time when medical intervention can be successful in reversing or slowing the rate of progression to kidney failure. Worse, even in its early stages, kidney disease poses significant cardiovascular risk; indeed, individuals with advanced CKD are more likely to die of cardiovascular disease than live long enough to need kidney replacement therapy. PMID- 19806866 TI - Lower eyelid reconstruction following Mohs surgery. AB - Lower eyelid defects resulting from Mohs micrographic surgery can be challenging to repair. These repairs are fraught with potential complication due to the lower eyelid's complex anatomy and defect variability. A single "cookie-cutter" treatment regimen does not exist because patients and defects vary. Surgical closure techniques include primary closure, eyelid advancement, rotational flaps, full thickness skin grafts, and/or allografts. We present a discussion of lower eyelid reconstruction including relevant anatomy, physical signs, and treatment options with examples. PMID- 19806868 TI - Caudal epidural blood patch. AB - This report describes the use of a single shot, through the needle caudal approach to epidural blood patch (EBP) in a patient with persistent leakage of cerebrospinal fluid following lumbosacral laminectomy. A previous report of caudal EBP in an adult patient with an epidural catheter suggested that the success of the procedure could be comparable without the use of a catheter. This case report documents the success of through the needle caudal EBP in an adult patient. PMID- 19806867 TI - Leuconostoc spp sepsis in an extremely low birth weight infant: a case report and review of the literature. AB - A three week old extremely low birth weight (ELBW) infant infected by vancomycin resistant Leuconostoc spp is presented. Treatment with appropriate antibiotics was successful after the percutaneous inserted central catheter (PICC) was removed. The infection with Leuconostoc spp is rare but should be suspected when vancomycin-resistant organisms resembling streptococci are isolated. Previous pediatric case reports are also summarized and reviewed. PMID- 19806869 TI - Parathyroid FNA and hormone assay. AB - Primary hyperparathyroidism is a relatively common problem encountered in clinical endocrine practice. In most cases the diagnosis is relatively straightforward, however, when imaging studies fail to localize the parathyroid adenoma or hyperplasia, management can be challenging. We describe here such a case where the diagnosis was made by a novel method of analysis of parathyroid hormone levels in the needle wash obtained during fine-needle aspiration of a suspected parathyroid adenoma. A 60 year old white male was first seen in the endocrinology clinic for evaluation of osteoporosis. He had history of multiple compression vertebral fractures involving thoracic and lumbar vertebrae and fracture of right femoral neck following minimal trauma. He had high normal serum calcium and elevated urinary calcium levels. His parathyroid hormone level was within normal limits. Work-up for secondary causes of osteoporosis was unremarkable. He was started on hydrochlorthiazide therapy for a presumptive diagnosis of idiopathic hypercalciuria. Subsequently his serum calcium level became elevated and he continued to have significant hypercalciuria. The elevation in serum calcium persisted despite cessation of hydrochlorthiazide therapy. Parathyroid hormone level remained in mid-normal range. A diagnosis of primary hyperparathyroidism was considered at this stage and imaging studies were carried out to localize the parathyroid pathology. Parathyroid-sestamibi scan did not reveal any abnormality. Ultrasound examination of the neck showed a hypoechoic nodule posterior to right thyroid lobe. A fine needle aspiration of the nodule was carried out with estimation of parathyroid hormone level in the needle wash to indicate the presence of parathyroid adenoma. This was surgically removed later successfully with subsequent normalization of serum and urinary calcium levels. The current management of hyperparathyroidism is primarily surgical. Minimally invasive parathyroid surgery is the treatment of choice but it requires the clear localization of a parathyroid lesion for successful removal. In cases where preoperative localization is evasive, novel techniques, such as the one described above, can provide useful diagnostic information which can aid in the successful management of hyperparathyroidism. Further studies are needed before this technique can be applied on a more widespread basis. PMID- 19806870 TI - Impairment by psychiatric disorders, including alcoholism and drug dependence. PMID- 19806871 TI - Revised federal guidance for nursing facility medical directors. PMID- 19806872 TI - Know the facts before you choose a DI policy. PMID- 19806873 TI - Health reform could get you hired. PMID- 19806874 TI - 10 tips for living to 100. PMID- 19806875 TI - Your guide to maximizing Medicare. PMID- 19806876 TI - Workout wisdom for those over 40. PMID- 19806877 TI - How to feel good in bad times. PMID- 19806878 TI - Building in China. Nashville-based firm backed by Frist strikes deal with Chinese government. PMID- 19806879 TI - Rapier capitalizes. WellMed doc receives exec award. PMID- 19806880 TI - By the numbers. 25 largest post-acute-care companies. Ranked by 2008 net revenue, based on Modern Healthcare's 2009 Post-Acute-Care Survey. PMID- 19806881 TI - The secrets inside your dog's mind. PMID- 19806882 TI - [Recent advances in study of antinociceptive conotoxins]. AB - The Conus venom is secreted by the duct and theca of venom. Most of conotoxins are composed of 10-40 amino acid residues with several disulfide bridges. They can specifically target neurotransmitter receptors including nAChRs, calcium ion channels, sodium ion channels and potassium ion channels, etc. Some conotoxins, such as that target N-Ca2+ channels, nAChR alpha9alpha10 subtype, TTX-R Na+ channels or NMDA receptors, have potent antinociceptive activities, omega-MVIIA, an Ca2+ channels blocker was approved by FDA in December, 2004 for marketing. Because of lower molecular weight and high specificity, conotoxins are the powerful pharmacology tools and potent analgesics without addiction. This review briefly summarizes the research progress of antinociceptive conotoxins and addresses on their targets and structure-activity relationships. PMID- 19806883 TI - [Research progress of virtual screening aided drug discovery]. AB - In the process of new drug discovery, the application of virtual screening can enrich active compounds, reduce the cost of drug screening, and increase the feasibility of drug screening. Therefore virtual screening technology has become an important approach for new drug discovery. As virtual screening and bioactivity screening possess different advantages, their combination can effectively promote new drug discovery. In the present paper, the application and the trend of removal of non-drug compounds, removal of false positive compounds, pharmacophore searching, molecular docking, and molecular similarity in the process of drug discovery are introduced in order to obtain more benefit from virtual screening strategy for new drug discovery. PMID- 19806884 TI - [A novel pulmonary delivery system--dry powder inhalers]. AB - Dry powder inhalers (DPIs) have received considerable attention because of their propellant-free composition and stability. DPIs include the DPI devices and inhalation powders. The purpose of this review is to address the development of the DPIs, including the mechanisms of absorption, the products, the devices, the preparation technology, and the characteristics. PMID- 19806885 TI - [Establishment of a diabetic-hypertensive rat model]. AB - This study is to establish a diabetic-hypertensive model in rats. After the induction of diabetes by streptozocin (STZ), rats were maintained with free access to rat chow and 1% NaCl drinking water. Blood pressure was monitored at conscious state by tail-cuff weekly till it was 50 mmHg higher than normal animal steadily. Finally, blood pressure was measured by catheterization of the right carotid artery and plasma ET-1 and Ang II, kidney Ang II and angiotensinogen or preproendothelin gene expression in liver or aorta were assayed separately. STZ diabetic rats that maintained with 1% NaCl drinking water exhibited obviously increasing blood pressure since the third week. Then the pressure reached 150 mmHg at the 6th week and was maintained until the 11th week. Till the 12th week, the blood pressure reached to higher than 160 mmHg. In addition, these high blood pressure rats were accompanied with increased blood plasma ET-1 and Ang II and augmented gene expression levels of angiotensinogen in kidneys and preproendothelin in aorta tissues. Loading sodium chloride chronically to STZ diabetic rats could prepare a diabetic-hypertensive rat model. PMID- 19806886 TI - [Effect of spinal glutamate transporter 1 on chronic constriction injury of sciatic nerve and morphine tolerance of rats]. AB - In order to investigate the role of spinal glutamate transporter 1 (GLT-1) in the neuropathic pain and morphine tolerance, rat chronic constriction injury (CCI) of sciatic nerve was performed, and the mechanical allodynia was evaluated by mechanical withdrawal threshold (MWT), the expression of GLT-1 was measured by real-time PCR and Western blotting analysis. The results showed that compared to sham group, the MWT of CCI group had decreased approximately 80%. Administration of morphine alone could develop tolerance rapidly in initial two days, and then had no significant difference with CCI group, the expression of GLT-1 was down regulated. Ceftriaxone sodium alone could improve mechanical allodynia. Co administration of ceftriaxone sodium with morphine attenuated morphine tolerance and up-regulated GLT-1 expression, and the MWT remained at high level after 6 days. In conclusion, change of spinal GLT-1 expression and function has close correlation with the development of neuropathic pain and morphine tolerance. PMID- 19806887 TI - [Protective effects of rhu TNFR: Fc against the lipopolysaccharide induced intestinal damage of rats and its underlying mechanism]. AB - To investigate the protective effects of recombinant human tumor necrosis factor receptor II: IgG Fc fusion protein (rhu TNFR: Fc) against the lipopolysaccharide (LPS) induced intestinal damage of rats and its underlying mechanism. SD rats were randomly divided into four groups: control group, rhuTNFR: Fc group, LPS group and rhu TNFR: Fc + LPS group. Mean arterial pressure (MAP) was continuously monitored and the mortality rates were assessed. The levels of TNF-alpha and its bioactivity in the serum were assessed by ELISA and flow cytometry respectively. Pathologic changes of intestinal tissue were observed by HE staining. The rats of control and rhu TNFR: Fc group all survived with stable MAP, and the low level and bioactivity of TNF-alpha in the serum were maintained. While 83% of the rats in LPS group died by 6 h with the levels and bioactivity of TNF-alpha increasing significantly. In rhu TNFR: Fc + LPS group, the mortality rate of rats dropped to 33%. The TNF-alpha level increased compared with control group but its bioactivity decreased significantly compared with LPS group. The MPO activity and content of MDA decreased significantly. The status of pathological manifestation in the intestine was also ameliorated. These data suggest that rhu TNFR: Fc could protect rats from the acute intestine injury induced by LPS through ablating the rise in serum TNF-alpha level and bioactivity as well as anti-oxidation. PMID- 19806888 TI - [Anti-proliferation action of taurine on rat cardiac fibroblast through inhibiting protein kinase Calpha expression]. AB - This project aimed to investigate the effect of taurine on cell cycle regulatory protein p27, Cyclin D1 and nuclear factor-kappa B (NF-kappaB) p65 in the proliferation of cultured neonatal rat cardiac fibroblast (CFb) induced by angiotensin II (Ang II), and to explore the effect of taurine on the signal transduction pathway in CFb proliferation. The cultured neonatal rat CFbs were isolated by trypsin digestion method. The proliferation of CFb was induced by Ang II and detected with thiazole blue (MTT) colorimetric assay. The protein expression of p-PKCalpha in cells was determined with Western blotting technology. The expression of p27 was analyzed by flow cytometry. The expression of Cyclin D1 was determined with the combination of immunocytochemical staining and image analysis software. The nuclear translocation of NF-kappaB p65 was determined with immunofluorescence staining. Among the concentrations ranged from 40 to 160 mmol L(-1), taurine significantly inhibited p-PKCalpha expression. Taurine increased p27 expression and inhibited the nuclear translocation of NF kappaB p65 in CFb (P < 0.05, P < 0.01, respectively) by inhibition of p-PKCalpha expression. And PKC inhibitor (Che) could improve the inhibitory action of taurine on CFb proliferation. The effects of taurine on CFb proliferation might be due to inhibition of p-PKCalpha expression and p27 expression increase and the nuclear translocation of NF-kappaB p65 inhibition followed. PMID- 19806889 TI - [Construction and screening of phage antibody libraries against epidermal growth factor receptor and soluble expression of single chain Fv]. AB - Recent studies have shown that epidermal growth factor receptor (EGFR) is an important target for cancer therapy. The present study prepared single chain Fv (scFv) directed against EGFR. Balb/c mice were immunized by human carcinoma A431 cells, and total RNA of the splenic cells was extracted. VH and VL gene fragments were amplified by RT-PCR and further joined into scFv gene with a linker, then scFv gene fragments were ligated into the phagemid vector pCANTAB 5E. The phagemid containing scFv were transformed into electro-competent E. coli TG1 cells. The recombinant phage antibody library was constructed through rescuing the transformed cells with help phage M13K07. The specified recombinant phages were enriched through 5 rounds of affinity panning and the anti-EGFR phage scFv clones were screened and identified with ELISA. A total of 48 clones from the library were selected randomly and 45 clones were identified positive. After infecting E. coli HB2151 cells with one positive clone, soluble recombinant antibodies about 27 kD were produced and located in the periplasm and the supernatant. The result of sequencing showed that the scFv gene was 768 bp, which encoded 256 amino acid residues. VH and VL including 3 CDRs and 4 FRs, respectively, were all homologous to mouse Ig. The soluble scFv showed the specific binding activity to purified EGFR and EGFR located in carcinoma cell membrane. The successful preparation of anti-EGFR scFv will provide an EGFR targeted molecule for the development of antibody-based drugs and biological therapy of cancer. PMID- 19806890 TI - [Synthesis and anti-tumor activities of N-substituted benzamide derivatives]. AB - To explore novel histone deacetylase (HDACs) inhibitors with anti-tumor activity, MS-275, a HDACs inhibitor, was prepared and used as a lead compound to design new N-substituted benzamide derivatives. MS-275 and eleven target compounds were obtained, and their structures were confirmed by 1H NMR and HR-MS individually. The results showed that the activity of compound 9d was equal to MS-275 in HDACs inhibition tests in vitro and worthy of further investigation. Compound 5c, 5d and 9c displayed obvious dose-effect relationship, which possessed moderate HDACs inhibitory activities. Ten compounds except 9e had selective inhibitory activities on Hut78. PMID- 19806891 TI - [The condensation mechanism of sodium new houttuyfonate and determination of the chemical structure of condensation products]. AB - To study the condensation mechanism of sodium new houttuyfonate, and determinate the chemical structure of condensation products, dimer was prepared, and LC-DAD MS/MS multiple techniques were employed to investigate the ultraviolet absorption feature and mass spectrum of transformation solution of dimer, and the transformation kinetics and half-life were studied by ultraviolet spectrophotometry. The pure substance of stable condensation product was obtained by extracting with organic solvent and purifying with column chromatography, the chemical structure of this substance was identified by assaying of IR, HR-ESI-MS and NMR, and the data of LC-MS/MS were compared with that of transformation products of dimer. The results indicated that the dimer is unstable, it will be rapidly dissociated in aqueous solution to form free new houttuyfonate and then cycloaddition reaction will occur and followed by an in situ dehydration to generate 1, 3, 5-tri (dodecanoyl) benzene (trimer) with a six-ring which is stable in aqueous solution. The transformation process may fit second-order kinetics, and the half-times were found to be 3.17 hours at 25 degrees C (298 K) and 6.39 min at 100 degrees C (373 K), separately. It suggests that dimer is an intermediate in condensation reaction, and the end condensation product of sodium new houttuyfonate injection may exist as trimer. PMID- 19806892 TI - [Anti-complement activity of polysaccharide B3-PS2 purified from Herba Scutellariae Barbatae]. AB - The polysaccharide B3-PS2 was extracted and purified from Herba Scutellariae Barbatae through chromatography of DEAE-cellulose and Sephacryl S-300 column. Average molecular weight of B3-PS2 was about 1,100 kD. It was composed of Glc, Gal and Ara in the ratio of 2.7:2.7:1.0, along with trace of Man, Rha, Fuc and Xyl. B3-PS2 inhibited complement activation on the classic pathways with CH50 value of (0.23 +/- 0.03) mg mL(-1). The targets of B3-PS2 upon the complement system were C1r, C1s, C3 and C4. These results suggested that anti-complementary activity of B3-PS2 was closed to its positive control heparin. It strongly suggested that the polysaccharide B3-PS2 from Herba Scutellariae Barbatae could be a potential candidate in treating those complement-associated diseases. PMID- 19806893 TI - Antifungal active triterpene glycosides from sea cucumber Holothuria scabra. AB - To study the new antifungal active triterpene glycosides of sea cucumber Holothuria scabra. Triterpene glycosides from Holothuria scabra were separated and purified by silica gel chromatography, reversed-phase silica gel chromatography and RP-HPLC. Their structures were elucidated on the basis of spectral data and chemical evidence. Three triterpene glycosides were identified as scabraside A (1), echinoidea A (2) and holothurin A1 (3). Scabraside A (1) is a new triterpene glycoside, and compounds 2 and 3 were isolated from Holothuria scabra for the first time. They showed antifungal activities (1 < or = MIC80 < or = 16 microg mL(-1)). PMID- 19806894 TI - [Triterpenes from fruits of Paliurus ramosissimus]. AB - To study the chemical constituents of the fruits of Paliurus ramosissimus, various chromatographic techniques were used to separate and purify the chemical constituents. Three triterpenes have been isolated and purified by using various column chromatography. Their structures were elucidated by their physico-chemical properties and spectroscopic data. These compounds were determined as: 22S, 23R epoxy-tirucalla-7-ene-3alpha,24, 25-triol (1), 21S, 23R-epoxy-21, 24S, 25 trihydroxy-apotirucalla-7-ene-3-one (2), 21R, 23R-epoxy-21-ethoxy-24S, 25 dihydroxy-apotirucalla-7-ene-3-one (3), separately. Compound 1 is a new compound, and the others were obtained from this genus for the first time. PMID- 19806895 TI - [Determination of lipase activity by gas chromatography]. AB - A rapid gas chromatography method was developed for determination of lipase activity using tributyrin as substrate. The standard curves of butyric acid hydrolyzed from tributyrin were linear in the range of 0.11-11.35 mmol L(-1). The recoveries of low, moderate and high concentrations of tributyrin were 90.3%, 104.6%, 89.4% with RSD of 3.01%, 4.50%, 6.64%, respectively. The incubation time was only 5 minutes which was less than with the half time of the conventional titrimetry and spectrophotometry. The optimum pH value was 7.5 and the optimum temperature was 32 degrees C. Based on the Lineweaver-Burk plots, the Michaelis Menten constant was 0.25 mmol mL(-1). The effect of orlistat on the enzyme inhibiting activity was studied to prove the accuracy of this method. It was found that the half-inhibition concentration (IC50) of orlistat was 0.0485 mg mL( 1). The small total reaction volume, the simple treating procedures, the high accuracy and precision present the advantages of the new method. PMID- 19806896 TI - [Effect of cyclosporine A on the pharmacokinetics of ginkgolide B in rats]. AB - The paper is aimed to investigate the effect of cyclosporine A (CyA) on the pharmacokinetics of ginkgolide B (GB) in rats, and to look for the mechanism of the changes in pharmacokinetic behaviors of GB. GB concentration in plasma, brain homogenate and urine samples of rats was determined by LC-MS. Effects of CyA on plasma levels, brain distributions as well as urinary excretions after intravenous administration of GB were evaluated. CyA co administrated intravenously at 10 mg kg(-1) or 20 mg kg(-1) significantly increased AUC(0-360 min) (P < 0.01) and decreased total CL of GB in rats. While co administrated CYP3A inhibitor itraconazole (ICZ) has no appreciable effect on the pharmacokinetic behavior of GB. CyA increased the brain uptake of GB in a dose dependent manner. The brain distribution of GB was significantly increased at 5 min by different doses of CyA (P < 0.001), while at 20 and 60 min only high dose of CyA could significantly increase the levels of GB in the brain (P < 0.01 and P < 0.001). Different P-gp inhibitors CyA or verapamil (VER) or digoxin (DGX) decreased the urinary GB excretion, the urinary excretion of GB in 0-8 h were about 34.8% (P < 0.001), 59.4% (P < 0.001) and 79.7% (P < 0.05) of the control, separately. No appreciable effect of ICZ was observed on urinary excretion of GB. Coadministration of P-gp inhibitors CyA could significantly increase the plasma level, accelerate the brain distribution and decrease the urinary excretion of GB. PMID- 19806897 TI - [Investigation of metabolic action of Cordyceps sinensis and its cultured mycelia on Escherichia coli by microcalorimetry]. AB - This study is to investigate the effect of Cordyceps sinensis and its cultured mycelia on growth and metabolism of Escherichia coli, and microcalorimetric method was carried out to evaluate its biological activity. The study will provide the basis for the quality control of Cordyceps sinensis. Experimental result will show the effect of natural Cordyceps sinensis and its cultured mycelia on growth and metabolism of Escherichia coli, with index of P(1max) and effective rate (E) by microcalorimetry, the data of experiment were studied by cluster analysis. The results showed that Cordyceps sinensis and its cultured mycelia not only can promote growth and metabolism of Escherichia coli but also can regulate the balance of intestinal microecology efficiently. When the concentrations of samples > 6.0 mg mL(-1), natural Cordyceps sinensis can promote the growth and metabolism of Escherichia coli efficiently (P < 0.05) compared with the control group, and have better dose-effect relationship with concentration (r > 0.9), its cultured mycelia does not show conspicuous auxoaction (P > 0.05) and have not dose-effect relationship with concentration (r < 0.6); when the concentration of samples < 6.0 mg mL(-1), all samples does not show conspicuous auxoaction (P > 0.05). Natural Cordyceps sinensis and its cultured mycelia can be distinguished by cluster analysis. Microcalorimetry has a good prospect on the quality evaluation of the traditional Chinese medicine. PMID- 19806898 TI - [Influence of combination of extractum Angelicae Dahuricae Siccum and total alkaloids of Rhizoma Corydalis on pharmacokinetics of tetrahydropalmatine in rats]. AB - This paper is aimed to develop a rapid and sensitive HPLC-fluorescence detection (FLD) method for the determination of tetrahydropalmatine (TET) in rats' plasma. The influence of combinations of Extractum Angelicae Dahuricae Siccum (coumarin and volatile oil) and total alkaloids (TA) from Rhizoma Corydalis (TA) on pharmacokinetics of TET in rats was studied. Plasma samples were treated with hexane-isopropanol (95:5) to precipitate the protein, and were determined by HPLC fluorescence detection. The calibration curve was linear in the range of 2.096 167.68 microg L(-1). The limit of quantification was 2.096 microg L(-1). The method recovery of TET was 94.0%-100.0%. The extract recovery was 72.0%-81.5%. RSDs ofintra- and inter-day precisions were all less than 7.0%. Pharmacokinetics of TET in rats was fitted to two compartments open model after oral administration of TA, TA-volatile oil (VO), TA-coumarin (Cou) and TA-VO-Cou. Compared with TA, AUC(0-t), AUC(0-infinity), MRT(0-t), and MRT(0-infinity) of TET had significant deviation when combined with VO and/or Cou. The determination method is sensitive, specific, accurate, and appropriate for determination of TET in vivo. Coumarin and/or VO combined with TA can prolong the resistance time of TET significantly, delay elimination and enhance bioavailability of tetrahydropalmatine. PMID- 19806899 TI - [Preparation of silymarin-loaded amphiphilic chitosan micelle and its in situ absorption in rat intestine]. AB - To improve the oral bioavailability of silymarin, the silymarin-loaded amphiphilic chitosan micelles (SM-OGC) were prepared. The absorption of SM-OGC in rat intestine was investigated. SM-OGC was prepared by dialysis method. The size and zeta potential of SM-OGC were investigated. Compared to silymarin suspension, the absorption of SM-OGC was investigated using in situ single pass perfusion model. The diameters and zeta potential SM-OGC were (162.4 +/- 3.0) nm and (+32.6 +/- 0.98) mV, respectively. The encapsulation efficiency was (39.17 +/- 0.98)% and the drug loading of SM-OGC was (28.15 +/- 0.43)%. The absorption of SM-OGC at different segments of intestine was significantly higher than that of silymarin suspension (P < 0.05). The apparent absorption rate (K(a)) and effective permeation coefficient (P(eff)) at the duodenum were the largest. K(a) and P(eff) had no significant difference between jejunum, ileum and colon. OGC micelles might significantly promote the absorption of silymarin in the intestine tract. PMID- 19806900 TI - Comparison of self-microemulsifying drug delivery system versus solid dispersion technology used in the improvement of dissolution rate and bioavailability of vinpocetine. AB - The objective of this study is to compare the differences between self microemulsifying drug delivery system (SMEDDS) and solid dispersion (SD) technology used to improve the dissolution rate and bioavailability of vinpocetine (VIP). The formulation of VIP-SMEDDS was composed of Labrafac, oleic acid, Cremophor EL, Transcutol P, and gum acacia which was used as solid absorbent. VIP-SD was prepared using poloxamer F68 as the carrier. In the solubility test, the solubility of VIP in SMEDDS was 17.3 times as much as that in SD. In the dissolution test, SMEDDS had shown better enhancement and stability in dissolving VIP than SD. When compared to VIP crude powder, the bioavailability of VIP in SMEDDS (VIP-SMEDDS) was 1.89-fold higher, and was less affected by food intake. However, the bioavailability of VIP in SD (VIP-SD) was bioequivalent to that of VIP crude powder. The tissue uptake of VIP-SMEDDS in Peyer's patches, intestine and liver after administration for 2 hours was more favorable than that of VIP-SD, which was 3.7 times higher in Peyer's patches, 2.2 times higher in intestine and 1.5 times higher in liver. In Caco-2 tests, the apparent permeability (P(app)) of VIP-SMEDDS was 2.65 times of that of VIP-SD. The width of the cell tight junctions of Caco-2 cell monolayer treated with VIP-SMEDDS were 9.6-fold wider, but there was no significant change after treatment with VIP-SD, when compared to the blank control. In conclusion, SMEDDS was more efficient than the traditional SD technology in increasing solubility, dissolution, intestinal permeability, lymphatic absorption and bioavailability of the insoluble drugs such as VIP, which is less affected by food intake. PMID- 19806901 TI - [Formation and aggregation behavior of polyethyleneimine-DNA complexes]. AB - In vitro gene delivery, polyethyleneimine (PEI) has been described as one of the most efficient nonviral vector. Herein the formation mechanism of PEI/DNA complexes is elucidated. The transition phase of "bead-on-string" structure in the formation of complexes was supposed to exist through spectroscopy, electrophoresis and transmission electron microscopy (TEM) technology. The construction of PEI/DNA complexes is related closely to the characteristics of PEI and DNA plasmid. As well as the dominant electrostatic effects, the nonelectrostatic interactions were thought to be partially responsible for the presence of PEI/DNA complexes even in the high ionic strength. The surface charge of complexes particles increased with the N/P ratio, but the absolute value of zeta potential was lower at the N/P ratio of 8 and 12, perhaps attributed to the use of larger DNA plasmid. As a result, the repulsion between particles was decreased and prone to aggregate to the structure like a clustered grape-string in the solution. Interestingly, contrast to the formation behavior of complexes, the PEI/DNA complexes aggregated primarily due to hydrophobic interactions while electrostatic attractions play a little role in the complexes particles aggregation in different concentrations of salt solutions. Comparable transfection efficiency in HepG2 cells was observed for the Lipofectamine 2000 and PEI/DNA complexes at the N/P ratio of 12, and showed that larger or aggregable complexes could transfect the cells in some different mechanisms. PMID- 19806902 TI - Pharmacokinetic modelling of microencapsulated metronidazole. AB - The aim of present study is to develop a pharmacokinetic model for microencapsulated metronidazole to predict drug absorption pattern in healthy human and validate this model internally. Metronidazole was microencapsulated into ethylcellulose shells followed by the conversion of these microcapsules into tablets. Dissolution study of tablets was conducted in 450 mL double distilled water, 0.1 mol L(-1) HCl and phosphate buffer (pH 6.8) maintained at (37+/-0.5) degrees C using USP apparatus II at 50, 100 and 150 r min(-1). Three metronidazole tablets (T1: fast release, T2: moderate release, T3: slow release and reference) were administered to twenty four healthy human volunteers and serial blood samples were collected for 12 hours followed by their analysis using RP-HPLC. Drug release data were analyzed by various model dependent and independent approaches. Drug absorbed (%) was determined by Wagner-Nelson method from plasma concentration profile. Internal predictability was checked from Cmax and AUC. Optimum dissolution profile was observed in double distilled water and 50 r min(-1). A good level A correlation was observed between drug dissolution and absorption profiles (correlation coefficient, R2 = 0.9009, 0.9426, 0.9015 and 0.932 for T1, T2, T3 and reference, respectively). Internal predictability was found less than 10%. Good correlation coefficients and low prediction errors elaborate the validity of this mathematical in-vitro in-vivo correlation model as a predictive tool for the determination of pharmacokinetics from dissolution data. PMID- 19806903 TI - [Genetic relationships of Atractylodes plants]. AB - The phylogenetic relationships of the genus Atractylodes DC. was analyzed using the nuclear ribosomal ITS and three chloroplast fragments, including atpB-rbcL, psbB-psbF and trnL-trnF intergenic spacer (IGS) sequences. Phylogenetic analyses revealed that A. lancea subsp. luotianensis and A. lancea, A. chinensis var. liaotungensis and A. coreana form monophyletic terminal clade, separately. The trees, within each the pairwise genetic distances, did not support Hu's classification. Authors suggested that A. lancea Subsp. luotianensis should be included in A. lancea rather than be treated as a separate subspecies. A. carlinoides was placed in the basal position of Atractylodes, which had a distant relationship with the others of the genus. The results lead us to suggest that A. chinensis var. liaotungensis be put into A. coreana, A. chinensis as a subspecies of A. lancea. PMID- 19806904 TI - [Caenorhabditis elegans: a powerful tool for drug discovery]. AB - A simple model organism Caenorhabditis elegans has contributed substantially to the fundamental researches in biology. In an era of functional genomics, nematode worm has been developed into a multi-purpose tool that can be exploited to identify disease-causing or disease-associated genes, validate potential drug targets. This, coupled with its genetic amenability, low cost experimental manipulation and compatibility with high throughput screening in an intact physiological condition, makes the model organism into an effective toolbox for drug discovery. This review shows the unique features of C. elegans, how it can play a valuable role in our understanding of the molecular mechanism of human diseases and finding drug leads in drug development process. PMID- 19806905 TI - [Recent advances in the study of bioreductive drugs targeted tumor hypoxia]. AB - Tumor hypoxia is the necessary process in the development of solid tumors, which is the key factor for drug resistance, recurrence, attack and shift of tumor. Hypoxic tumor cells have a certain extent of tolerance to radiation and chemotherapy. Tumor hypoxia is an important target for medication therapy. In the recent years, the bioreductive drugs targeted tumor hypoxia has made great process in the treatment of tumors. The latest advances of bioreductive drugs targeted hypoxia were reviewed in this paper. PMID- 19806906 TI - [Oral recombinant adeno-associated virus gene medicine]. AB - The efficacy of recombinant adeno-associated virus (rAAV) vector-mediated gene delivery to the gastrointestinal tract has been paid a considerable attention over the last 10 years, since our first report on the oral gene pill strategy in Nature Medicine, even though there are still several potential obstacles for this route to overcome in order to obtain efficient gene delivery. The preclinical results of oral rAAV gene medicine are summarized in this review, and special attention is paid on its pharmacokinetic and pharmacodynamic aspects with an emphasis on drug delivery, absorption, distribution and transduction. The rAAV based vectors have been shown promising results in human clinical trials with fewer safety concerns over other gene medicines. However, the underlying mechanisms and biopharmaceutical features of oral rAAV gene medicine remain to be explored extensively and intensively to develop this novel technology as a treatment for a wider range of diseases. PMID- 19806907 TI - [Progress in the study of micelle delivery system reversing multidrug resistance]. AB - Multidrug resistance (MDR) of cancer cells to anti-tumor drugs remains a major impediment to successful chemotherapy. There has been an increasing interest in the studies of the mechanism and reverse of the MDR. Being a reliable and safe way to reverse MDR, drug delivery systems (DDS) such as micelle, liposome and nanoparticle, represent a promising prospect both in research and application in recent years. On the basis of recent studies, the effect and mechanism of micelles on reversing MDR are reviewed. And it is anticipated that DDS could contribute greatly to reversing MDR in the future. PMID- 19806908 TI - [Neonatal fluoxetine exposure induced depression-like behaviors in adult Kunming mice and the antidepressant-like effect of agmatine]. AB - This study is to explore a behavioral and pathological model for depression in mice, and evaluate the anti-depressant-like effect of agmatine. Neonatal Kunming mice were treated with fluoxetine (10 mg x kg(-1), ip, qd) for 17 d (between day 4 and 21 after birth), and then the mice were normally housed till being adult (about 10 weeks after birth). The behaviors of the mice were measured by using open-field test, novelty suppressed feeding test and tail-suspension test. Hippocampal adenylate cyclase (AC) activity was measured by radioimmunoassay. Neonatal exposure to fluoxetine induced a "depression-like" behaviors in the adult mice, shown as the decreased locomotor activity, increased feeding latency and immobility time in the open-field test, novelty suppressed feeding test, and tail-suspension test, respectively. Chronic agmatine treatment (10 mg x kg(-1), ig, bid) for 3 weeks significantly increased the locomotor activity, and decreased the feeding latency in the neonatal fluoxetine exposed mice. Furthermore, single treatment with agmatine (40 mg x kg(-1), ig) also decreased the immobility time in the tail-suspension test, and increased the hippocampal AC activity in the mice. These results indicate that neonatal exposure to fluoxetine induces depressive-like behaviors in the adult mice. Agmatine reverses these behaviors, which may be closely related to the enhancement of the hippocampal AC activity. PMID- 19806909 TI - [Pharmacokinetics of 6beta-naltrexol after single and multiple intramuscular injections in Beagle dogs]. AB - The pharmacokinetics of 6beta-naltrexol (6beta-NOL) following single intramuscular administration and multiple intramuscular injection once per day for seven days was studied in 4 Beagle dogs. Plasma concentration of 6beta-NOL in dogs was analyzed by a combination of high performance liquid chromatography (HPLC) and electrochemical detection with naloxone (NLX) as internal standard. After single intramuscular injection of 0.2 mg x kg(-1) 6beta-NOL, the plasma concentration-time curve of the drug was found to fit to a two compartment model with first-order absorption. The main parameters of single dosing were as follows: t1/2alpha was (0.26 +/- 0.23) h, t1/2beta was (4.77 +/- 1.65) h, C(max) was (81.65 +/- 5.61) ng x mL(-1), t(peak) was (0.27 +/- 0.07) h, CL(s) was (1.20 +/- 0.06) L x kg(-1) x h(-1), V/F(c) was (1.94 +/- 0.15) L x kg(-1), and AUC(0-t) was (166.82 +/- 7.68) ng x h x mL(-1), separately. After multiple intramuscular injection of 0.2 mg x kg(-1) 6beta-NOL once per day for seven days, the plasma concentration-time curve of the drug fitted to a two compartment model with first order absorption too. The main parameters of the last dosing were as follows: t1/2alpha was (0.19 +/- 0.18) h, t1/2beta was (5.79 +/- 1.50) h, C(max) was (79.82 +/- 10.5) ng x mL(-1), t(peak) was (0.18 +/- 0.08) h, CL(s) was (1.12 +/- 0.07) L x kg(-1) x h(-1), V/F(c) was (2.10 +/- 0.27) L x kg(-1), and AUC(0-t) was (173.23 +/- 9.49) ng x h x mL(-1), separately. The difference of the parameters between the first and the last dosing was not significant, showing that the plasma kinetics of 6beta-naltrexol was not changed after multiple administrations. In the course of multiple administration, the peak and valley concentration of plasma 6beta-naltrexol were (79.03 +/- 10.3) and (1.50 +/- 0.93) ng x mL(-1), respectively. No clear adverse events were noted during this study. These results showed that plasma 6beta-naltrexol fits to a two compartment model with first-order absorption in dog after intramuscular administration and their pharmacokinetic parameters were reported. There was no remarkable change on plasma pharmacokinetics of 6beta-naltrexol after multiple intramuscular administrations. PMID- 19806910 TI - [Structure-activity relationships analysis of thienorphine and its derivatives]. AB - Thienorphine is a chemically-new opioid developed in Beijing Institute of Pharmacology and Toxicology. To elucidate the chemical basis for the unique pharmacological effects of thienorphine, 15 derivatives were synthesized according to combinatorial chemistry and the structure-activity relationships of these compounds were studied. It is demonstrated that thienorphine is a potent long-acting partial agonist. N-Cyclopropylmethyl is responsible for the antagonist effect of thienorphine. More importantly, thiophene at the end of side chain is most likely the pharmacophore accounts for the long-lasting effect of thienorphine. Change of the connection of thiophene and the side chain does not result in changes in the antinociceptive activity. PMID- 19806911 TI - [Regulation of Astragalus heteropolysaccharides on synoviocytes apoptosis and proinflammatory cytokine secretion of rats with adjuvant arthritis]. AB - This study is to observe anti-inflammation mechanism of Astragalus heteropolysaccharides (AHPS) on rats with adjuvant arthritis (AA). Rats were treated with AHPS (1 000, 500, and 250 mg x kg(-10, ig) and Tripterygium wilfordii polyglycolide (TWP, 60 mg x kg(-1), ig), separately. TNF-alpha and IL 1beta contents in serum were determined with radioimmunoassay, pathomorphologic changes of synovium of knee joint were observed by histological section with HE staining, synoviocyte apoptosis of knee joint of rats was analyzed by Tunel detection, and Bax and Bcl-2 positive expression were detected by immunohistochemical method. The results were as follows: (1) both AHPS and TWP could improve significantly primary and secondary clinical symptoms of rats with AA and inflammatory response in articular synovium; (2) the contents of TNF-alpha and IL-1beta in serum of rats with AA increased significantly composed with those in groups treated with AHPS (1 000 and 500 mg x kg(-1)), and the amount of synoviocyte apoptosis decreased significantly (P < 0.01 or P < 0.05); (3) the positive expression of Bax in synovium of rats with AA was a little bit higher than that in normal control (P > 0.05), but the positive expression of Bcl-2 significantly increased (P < 0.01). AHPS (1 000 and 500 mg x kg(-1)) could up regulate positive expression of Bax and down-regulate the positive expression of Bcl-2 significantly (P < 0.05 or P < 0.01). The results show that AHPS can evidently decrease TNF-alpha and IL-1beta level in serum of rats with AA, which is one of molecular mechanisms that AHPS has anti-inflammatory properties. AHPS can induce synoviocyte apoptosis of rats with AA, which is achieved by the regulating effect of AHPS on the positive expression of Bax and Bcl-2. PMID- 19806912 TI - [Effect of ibudilast on apoptosis of airway eosinophil in asthmatic guinea pigs]. AB - This study is to investigate the effect of ibudilast on apoptosis of airway eosinophil in asthmatic guinea pigs and its mechanism. Experimental asthma model of guinea pigs was induced with ovalbumin (OVA). Differential count in BALF was examined. The apoptosis of eosinophils (EOS) was labeled with TdT-mediated dUTP nick end labeling (TUNEL) technique. Fas mRNA expression of EOS was detected by reverse transcription-polymerase chain reaction (RT-PCR). The quantification of GM-CSF and IL-5 in BALF was conducted with ELISA. After treatment of ibudilast, the number of EOS and the quantification of GM-CSF and IL-5 decreased significantly. The number of apoptotic cells as well as Fas mRNA expression of EOS obviously increased. The results indicated that anti-asthma mechanisms of ibudilast can antagonize asthma through decreasing the number of EOS, inducing apoptosis of EOS, enhancing Fas mRNA expression of EOS and reducing the content of GM-CSF and IL-5. PMID- 19806913 TI - [Inhibitory effect of multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes on human colon cancer cells proliferation]. AB - In order to maximize the efficiency and versatility of the vector-based siRNA approach, we have developed a novel siRNA expression vector containing multiple tandem siRNA cassettes to investigate the synergistic inhibitory effect of it on human colon cancer cells proliferation. Multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes was constructed and transfected into human colon cancer cell. The effect of multiple siRNA recombinant expression vector was detected by RT-PCR, Western blotting and flow cytometry. It was confirmed by restriction endonuclease and sequence analysis that multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes was constructed successfully. Livin and Survivin genes inhibition ratio of Livin and Survivin siRNA at mRNA levels were 27.90% and 32.24%, at protein levels were 22.28% and 40.86%, the apoptotic ratio was (11.69 +/- 1.37) %, but the synergistic effect was weaker than Livin and Survivin RNA interference, respectively. The multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes has been constructed successfully. It can inhibit the expression of Livin and Survivin genes in human colon cancer cells, but the synergistic effect was weaker than Livin and Survivin RNA interferences, respectively. PMID- 19806914 TI - [Design and synthesis of 3'-methyl-furanonucleosides and their anti-tumor activities]. AB - Taking 3'-Me-Ado (3'-methyladenosine) and Cladribine as the leading compounds, seventeen 3'-C-methyl-furanonucleosides were designed and synthesized. All the structures were confirmed by 1H NMR and MS. The target compounds were tested in vitro against human pulmonary carcinoma A549, human colon carcinoma LOVO and human leukemia CEM by MTT assay. The results showed that these compounds possessed moderate cytotoxicities. PMID- 19806915 TI - [Synthesis and in vitro cytotoxicity of naphthalimide polyamine conjugates as antitumor agents]. AB - Six naphthalimide polyamine conjugates were synthesized and their structures were confirmed by elemental analysis, 1H NMR, 13C NMR and MS. Antitumor activities were evaluated in vitro using MTT assay on Leukemia cells (K562), human breast cancer cells (MB-231) and prostate cancer cells (Ln cap cell). The results showed that most of the six compounds were superior to the control (amonafide), 6d, 6e, and 6f exhibited nice selectivity in a screen of hepatoma cells (BEL-7402) and human normal hepatocytes (QSG-7701). PMID- 19806916 TI - [The feasibility of application of reverse docking method to the selectivity studies of protein kinase inhibitors]. AB - This investigation is to explore the feasibility of applying reverse docking method to the selectivity studies of protein kinase inhibitors. Firstly, a database that consists of 422 protein kinase structures was established through collecting the reported crystal structures or homology modeling. Then a reverse docking based method of protein kinase target screening was established, followed by the optimization of related parameters and scoring functions. Finally, seven typical selective kinase inhibitors were used to test the established method. The results show that the selective targets of these inhibitors have relatively high scoring function values (ranking in the first 35% of the tested kinase targets according to the scoring function values). This implies that the reverse docking method can be applied to the virtual screening of kinase targets and further to the selectivity studies of protein kinase inhibitors. PMID- 19806917 TI - [New homoisoflavanones from Polygonatum odoratum (Mill.) Druce]. AB - To study chemical constituents of Polygonatum odoratum (Mill.) Druce, the compounds were separated with column chromatography and HPLC. On the basis of physicochemical properties and spectral data, their structures were confirmed. Nine compounds were isolated and identified as 5,7-dihydroxy-6-methoxyl-8-methyl 3-(2',4'-dihydroxybenzyl)chroman-4-one (1), 5,7-dihydroxy-6-methyl-3-(2',4' dihydroxybenzyl)chroman-4-one (2), 5,7-dihydroxy-6-methoxyl-8-methyl-3-(4' methoxybenzyl)chroman-4-one (3), disporopsin (4), chrysoeriol (5), 5,4'-dihydroxy 7-methoxy-6-methylflavone (6), N-trans-feruloyltyramine (7), N-trans feruloyloctopamine (8), and (+)-syringaresinol (9). Compounds 1-3 are new homoisoflavanones. Compounds 4-9 are isolated from this plant for the first time. PMID- 19806918 TI - A new triterpene from the fruiting bodies of Ganoderma lucidum. AB - A new lanostanoid triterpene, named ganoderitriol M (1), together with a known triterpene ganoderic acid epsilon (2), were isolated from the fruiting bodies of G lucidum. Compound 1 was deduced as (24S)-lanosta-7-oxo-8-en-3beta, 24, 25-triol on the basis of spectral analysis (UV, IR, MS, 1H NMR, 13C NMR and 2D NMR). PMID- 19806919 TI - Establishment of HPLC-ESI-MS method for the determination of eplerenone in human plasma and its pharmacokinetics. AB - A sensitive high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method was established for the determination of eplerenone (EP) in human plasma. The plasma samples of EP were extracted with ethyl acetate and separated by HPLC on a reversed phase C18 column with a mobile phase of 10 mmol x L(-1) ammonium acetate water solution-methanol (30 : 70, v/v). EP was determined with electrospray ionization-mass spectrometry (ESI-MS) in the selected ion monitoring (SIM) mode. The calibration curves were linear over the range of 2-4 000 ng x mL(-1) for EP. The lower limit of quantification was 2 ng x mL(-1). The method has been successfully applied in the pharmacokinetic study of the EP tablets. The main pharmacokinetic parameters of EP after oral administration of 25 mg, 50 mg, 100 mg were as follows, t1/2: (4.9 +/- 2.1), (4.7 +/- 1.5), (5.9 +/- 1.2) h; AUC(0-infinity): (4 402 +/- 1 735), (8 150 +/- 2 509), (13 783 +/- 4 102) microg x h x L(-1); and MRT: (6.2 +/- 2.1), (6.6 +/- 1.3), and (7.2 +/- 1.6) h. Parameters of EP after oral administration of multiple doses of 50 mg were as follows, t1/2: (6.1 +/- 1.7) h; AUC(ss): (10 071 +/- 4220) microg x h x L(-1); MRT: (8.1 +/- 2.3) h; and DF: (3.2 +/- 1.0). PMID- 19806920 TI - [Cloning, expression and the application of human, rat alcohol dehydrogenase and aldo-keto reductase]. AB - This study is aimed to clone and express human, rat alcohol dehydrogenase (ADH) and aldo-keto reductase. Then the enantioselective metabolism of mandelic acid (MA) was studied. Human alcohol dehydrogenase 2, rat alcohol dehydrogenase 1, human and rat aldo-keto reductase 1A1 were amplified using RT-PCR from human and rat liver samples. Then subcloned into pET-28a (+) and expressed in E. coli BL21 (DE3) stably. The protein was induced with IPTG and purified by affinity chromatography. Then the enzyme activities were measured. MA enantiomers were incubated with rat, human ADH and phenylglyoxylic acid (PGA) with AKR1A1, respectively. The metabolism was analyzed with HPLC. The proper genes were cloned and purified and proteins were obtained. All of the proteins obtained showed good activity. Stereoselective-metabolism of MA was observed in human ADH2, which favors for S-MA metabolism. The expression plasmids are constructed and the recombinant proteins are expressed successfully. The recombinant alcohol dehydrogenase and aldo-keto reductase have been employed to study MA metabolism. PMID- 19806922 TI - [Preparation of adriamycin-loaded temperature/pH sensitive self-assembly block copolymer micelles]. AB - The dialysis method was employed to load adriamycin into the micelles formed by temperature and pH sensitive polyhistidine-co-DL-lactide-co-glycolide polyethylene glycol poly DL-lactide-co-glycolide-co-histidine (OLH-b-PLGA-b-PEG-b PLGA-b-OLH). The critical micelle concentration (CMC) of the copolymer was measured with pyrene fluorescent probe method under different temperatures. The entrapment rate and drug-loading rate were determined with dialysis method. The diameter, morphology and surface potential of the copolymer micelles were investigated by corresponding instruments, respectively. The release behavior of adriamycin from copolymer micelles and the pH sensitivity were studied. The CMC of the copolymers ranged from 0.022 4 to 0.001 7 microg x mL(-1). The entrapment rate and drug-loading rate were 92.8% and 15.7%, respectively. The micelles have a mean diameter of (61.7 +/- 13.4) nm, and zeta potential was -9.88 mV. The in vitro adriamycin release rate increased with the pH dropping from 7.4 to 5.0. The results indicated that the CMC of the copolymers decreased as the raising of temperature, drug release behavior from the micelles possessed clearly pH sensitivity, and the copolymers may have a potential in targeted delivery system for anticancer drugs. PMID- 19806921 TI - [Population pharmacokinetics research of clozapine in Chinese schizophrenic patients]. AB - The goal of this study is to investigate the population pharmacokinetics of oral given clozapine in Chinese schizophrenic patients and to identify possible relationships between population parameters and covariates including demography factors and CYP1A2 genetic polymorphism, so as to create the population pharmacokinetics model to guide individual clinical delivery. Details of drug dosage history, sampling time and concentration of 626 data points from 183 patients were collected retrospectively. The 183 patients were randomly allocated either to the index group (n = 168) or to the validation group (n = 15). Population pharmacokinetic data analysis was performed using the nonlinear mixed effects model (NONMEM) program on the index group. The values of apparent clearance (CL/F), apparent volume of distribution (V/F) and the constant of absorption rate were estimated. A number of covariates including demographic index, coadministration of other drugs and CYP1A2 genotypes were evaluated statistically for their influence on these parameters. The final population model related clearance with day-dose/BSA (DBSA) and smoke habit (SMOK). Predictive performance of the final model evaluated with the validation group showed insignificant bias between observed and model predicted concentrations. Typical value of CL/F (non-smoking group), V/F and the constant of absorption rate were 28.5 L x h(-1) (5.05%), 1 290 L (16.7%) and 2.26 h(-1) (fixed), inter-patient variability (CV) in CL/F and V/F was) 42.2% and 10.0%, respectively. It was observed that the values of CL/F in the two smoking groups were higher than that in the non-smoking group. The residual variability (SD) between observed and model-predicted concentrations was 45.8 microg x L(-1). PMID- 19806923 TI - [The pathway of absorption and conveying of puerarin microemulsion-in-oil]. AB - The best absorption location of puerarin microemulsion-in-oil in intestine parva of rat and pharmacokinetic characteristics, and the pathway of absorption and conveying of puerarin microemulsion were studied. In situ rat perfusion method was used to investigate the intestinal absorption of puerarin. Through the changes of drug concentration in blocked and unblocked lymphs, to determine the pathway of absorption and conveying. Puerarin microemulsion-in-oil can be absorbed in any part of intestine, and the K(a), P(app) of every part is ileum > duodenum > jejunum > colon, and the K(a), P(app) of ileum is significantly larger than that of others. The absorption rate of different concentrations is not significantly different (P > 0.05). The puerarin transited by gastrointestinal tract, about 36.8% is absorbed by the lymphatic channels to enter the systemic circulation and 63.2% is absorbed by the non-lymphatic channels. The best part of intestine to absorb puerarin microemulsion is ileum, and it is passive transport. The pathway of conveying is lymphoid and non-lymphoid transit. PMID- 19806924 TI - [Preparation of the oral self-microemulsifying drug delivery system of GBE50]. AB - To prepare the oral self-microemulsifying drug delivery system (SMEDDS) of GBE50, balance solubility method was used to screen emulsifier and assistant emulsifier; a pseudo-tamary phase diagram was used to prepare microemulsion; and orthogonal design was used to optimize formulation. Self-microemulsifying efficiency, dissolution, stability and pharmacokinetics of the preparation were studied. As a result, GBE50-SMEDDS of IPM, Cremophor EL, 1,2-propanediol and GBE50 could be self emulsified to form stable microemulsion with particle diameter between 20 and 50 nm when emulsifying with water. Its self-microemulsifying efficiency and dissolution are quick with good stability and it has a higher bioavailability than market existing agents Xingling particles. GBE50-SMEDDS is stable and effective. PMID- 19806925 TI - Molecular cloning and characterization of a novel isoflavone reductase-like gene (FcIRL) from high flavonoids-producing callus of Fagopyrum cymosum. AB - Lignans are important defensive compounds in plants and have good biological activities protecting human health. In order to study the medicinal secondary metabolism of Fagopyrum cymosum (Trev.) Meisn, a traditional Chinese medicine with anti-tumor effect, a novel isoflavone reductase-like gene, FcIRL, was cloned using RACE strategy from a cDNA library of high flavonoids-producing callus. The full-length cDNA of the FcIRL was 1 217 bp (accession no. EU116032), which contained a 942 bp open reading frame (ORF) encoding a 313 amino acid protein. Two stop codons (TAG) and a putative polyadenylation signal ATAAA at 24 bp upstream from the polyadenylation site was found in 5' and 3' UTR, separately. And no intron was found in the genomic sequence yet. FcIRL contained a predicted N-terminal acetylation site (M1-K5) and a NADPH-binding motif (G10-G-T-G13-Y-I G16) in the N-terminal region, a conserved NmrA (nitrogen metabolite repression regulator) domain (V6-N244), multi-phosphorylation sites and one conserved N glycosylation site (N214). Sequence homology comparison, phylogenetic analysis and advanced structures prediction all suggested that FcIRL belonged to the class of pinoresinol-lariciresinol reductase (PLR), which is a key enzyme in synthetic pathway of 8-8'-linked lignans, with function in catalyzing reduction of pinoresinol and lariciresinol into secoisolariciresinol, and medicinal secondary metabolism and resistance in F. cymosum. PMID- 19806926 TI - [Determination methods for erythropoietin receptor activator in human urine]. AB - In the present study, isoelectronic focusing with different pH gradients (pH 3-5, 2-6) or migrating distances (8.5, 12 and 17 cm) and SDS-PAGE was used to separate continuous erythropoietin receptor activator (CERA), recombinant human erythropoietin (rhEPO), darbepoetin and endogenous EPO spiked in human urine with 37 degrees C overnight incubation. Double blotting and chemiluminescent visualization were used to detect the IEF and SDS-PAGE profiles. The bands of CERA profile were detected and well separated from the endogenous EPO and the other two EPO preparations with both SDS-PAGE and the IEF method using a gradient pH 3-5 and a migrating distance of 17 cm, and a significant particular band of CERA profile was found in the IEF result. These preliminary results indicated that the methods were reliable and reproducible for detecting CERA, and could be used as a routine procedure for anti-doping analysis. PMID- 19806927 TI - Hard knocks. How to keep high school kids with concussions on the bench. PMID- 19806928 TI - Monitoring quality of care in residential care for the elderly: the information challenge. AB - This study assesses the administrative data compiled on residential care facilities for the elderly (RCFEs) by the state of California and considers the feasibility of their adaptation into a comprehensive information system. Required state RCFE reporting forms were reviewed for potential data elements. Recording and reporting variation was evaluated using a stratified probability sample of 340 facilities licensed in Northern and Central California. Stratification was by facility size and state district office. Data collection included a 5-year retrospective review of forms and documents in each facility's public file. Little of the information required from RCFEs is computerized. Most of it is maintained at the individual facility and not included in public files. Basic information, while included in the public file, is commonly either not available or not current. Resident characteristics and outcomes are not compiled, except indirectly in citations. The information required from RCFEs, if appropriately compiled and maintained, would produce a comprehensive quality assurance system and more effectively support consumer information and policy needs. PMID- 19806930 TI - We've looked at care from both sides now: the effects of alternative evaluation strategies on study conclusions. AB - This study uses two studies about the role of managed-care programs in serving Medicaid long-term care clients in Florida to illustrate how different research designs can reach divergent conclusions. Two reports from different groups using essentially the same database to assess the impact of managed care on a group of older Medicaid clients served by a Nursing Home Diversion Program reached different conclusions. The report from Florida's Office of Program Policy Analysis and Government Accountability concluded that the Diversion program saved money, whereas the report from the Florida Policy Exchange Center on Aging at the University of South Florida reached basically the opposite conclusion. Both agreed that the capitation rate was too high. How the policy questions are framed and analyzed can affect the conclusions reached. A variety of factors can influence the apparent effects of programmatic interventions. Evaluations must take relevant confounding variables into account. PMID- 19806929 TI - Commentary: is assisted living in the United States well served by regulations requiring the reporting of detailed operational data that are then posted on the Internet? PMID- 19806932 TI - The power of silver: age and identity politics in the 21st century. AB - Pensioner political movements emerged in the interwar years in America and Europe. Documentary and empirical analyses confirm the influential role such movements played in helping shape the postwar social security systems of Western societies. Pensioner movements, qua pensioner movements, have failed to retain their influence, despite that "old age" and its demographic significance have become more salient. We propose three explanations for this: the first concerns the failure of old age to connect with the generational ethos of identity politics; the second reflects the nature of the actors now involved in the governance of old age; and the third concerns the individualization of retirement as a phase of life. PMID- 19806931 TI - What happens to defined contribution accounts when labor markets and financial markets move together? AB - The relationship among earnings, savings, and retirement is well known; however, the linkage between labor market outcomes and financial market performance is generally unacknowledged. We examine the implications of the link between labor markets and financial markets for workers who save money in individual retirement accounts. Specifically, differences in labor market outcomes across groups may imply differences in the timing of investments, which may reduce savings over time for these groups compared to their counterparts. Using monthly data from the Current Population Survey (1979-2002), we generate hypothetical investment portfolios using stock and bond indices. We exploit differences across demographic groups in unemployment and wage growth and use these differences to examine each group's investment outcomes. We then disaggregate the total effects into short-term and long-term components. We find some evidence of short-term market timing effects on investment, but we find much larger long-term effects for some groups. Our findings suggest that, for many people, the retirement savings losses associated with the timing of markets are similar to the costs of annuitizing savings upon retirement. The differences are especially pronounced by education and gender. PMID- 19806933 TI - [Reflection of the evidence-based medicine's principles in the Russian consensus documents on angiology and vascular surgery]. AB - Analysed herein are the consensus documents concerning angiology and vascular surgery worked out and adopted in Russia. This is followed by a detailed description of the methodology used while their development, underlying their significance for medical practice and pointing out their strict compliance with the principles of evidence-based medicine. Emphasis is laid on the thesis that the consensus guidelines worked out by the leading specialists, like any rules and norms, contribute favourably to making appropriate decisions by the practicing clinical physicians and may serve as a basis for creation of the branch standards. Also shown are advantages and shortcomings of the adopted documents in angiology, including phlebology, and finally announced is a forthcoming meeting of the Russian experts, devoted to the development of the consensus document entitled "Russian Clinical Guidelines on Diagnosis and Treatment of Chronic Venous Diseases". PMID- 19806934 TI - [Haemorheological problems in lower-limb chronic arterial insufficiency]. AB - Chair of Neurology of the Advanced Medical Training Department of the Federal Educational Facility of the Higher Vocational Education Russian State Medical University under the Russian Public Health Ministry, Moscow, Russia. The article deals with the findings obtained in examining and treating a total of eighty seven patients suffering from atherosclerosis obliterans of the lower-limb arteries with stage IIB and III chronic arterial insufficiency. Studied were impairments of the theological properties of the blood and plasma, erythrocytic deformability and the concentration of malonic dialdehyde in blood at admission to the hospital, upon completion of the course of conservative treatment, and after the reconstructive vascular intervention performed. It was determined that the leading mechanism disturbing oxygen utilization by tissues is a decrease in erythrocytic deformability, which promotes shunting of the arterial blood flow and persists after the surgical vascular reconstruction preformed. PMID- 19806935 TI - [Haemodynamic role of blood-plasma circulating cell-free DNA and contained therein high-molecular-weight CpG-rich fraction in pathogenesis of arterial hypertension and atherosclerosis obliterans of carotid arteries]. AB - The hydrodynamic resistance (HR) of blood is one of the components of the total peripheral resistance. High-molecular-weight DNA appears to decrease the HR in accordance with the Toms's effect. The present study was undertaken to investigate the HR and properties of cell-free DNA circulating in the blood plasma (hereinafter referred to as pDNA) of the control donors, patients suffering from either arterial hypertension (AH) alone or that combined with atherosclerotic lesions of the carotid arteries (CAs). Within the normal concentrations of pDNA, we revealed an inverse dependence of the HR thereupon and upon the content in pDNA of the high-molecular-weight CpG-rich fraction (CpG DNA), i. e., a transcribed region of the ribosomal repeat (rDNA). A decrease or an increase in the pDNA concentration in all the patients examined was accompanied by an elevation of the rDNA concentration in the blood plasma. Exceeding a certain level thereof appeared to give rise to an increase in both the HR and arterial pressure (AP). Patients presenting with degree I essential AH were found to have a decreased endonuclease activity of the blood plasma, with the pDNA concentration being more than two-fold higher with no change in the rDNA content. Their HR appeared to be increased (p<0.01). Patients diagnosed as having degree II AH were characterized by a normal or decreased level of pDNA and an elevated content of pDNA, with the HR being slightly lowered. In patients presenting with atherosclerosis obliterans of the ACs, the initial manifestations of the lesions of the carotid arteries were typically revealed on the background of a lowered HR (p<0.05). All patients suffering from atherosclerotic lesions of the ACs could be subdivided into two groups, which in our opinion is probably associated with different various mechanisms of destructive damage to the arterial intima. In some of them, the pDNA concentration does not differ from the normal values, but in its composition, there is an increased content of rDNA, elevating as obliteration of the vessels' lumen increases, with the HR being decreased. The majority of them have degree II AH. In others, the pDNA concentration is by an order of magnitude higher than the normal values, while the rDNA content in pDNA is decreased, with the HR being elevated. Most of them have degree III AH. Pronounced and rough stenoses take an asymptomatic course in patients with decreased values of the HR and a slightly elevated level of pDNA and/or rDNA in the blood plasma. A higher level thereof leads to a rise in the HR and to the appearance of neurological symptomatology. Hence, CpG-DNA circulating in the composition of pDNA is a constantly acting endogenous blood factor decreasing the HR (the Toms's effect) and normalizing AP under physiological conditions, being however a cause of their increase and impairment of blood circulation in the pathogenesis of AH and atherosclerosis obliterans of the CAs. PMID- 19806936 TI - [Mediator interactions in vasculogenesis and angiogenesis]. AB - The article contains a review of the literature dedicated to the problems concerning mediator interactions in vasculogenesis and angiogenesis. Also presented herein is the role of the key angiogenic mediators and growth factors in the processes concerned, as well as certain variants of vasculo- and angiogenesis impairments associated with mediators dysfunction discussed. PMID- 19806937 TI - [Study of the microbial spectrum of ulcerative-and-necrotic lesions in gerontological patients with lower limb chronic ischaemia]. AB - The present work deals with a comprehensive study of the microbial spectrum of an ischaemic trophic ulcer in gerontological patients with degree IV lower limb chronic ischaemia in order to improve therapeutic outcomes by means of selecting individual antibacterial therapy for a possible prognosis of the results of treatment for an ulcerative-and-necrotic lesion of distal portions of the lower extremities depending on the spectrum of the microflora vegetating in the wound. Based on examining a total of 130 patients aged 70 years and older suffering from lower limb chronic critical ischaemia, we revealed that the pathogen of secondary wound infection isolated in trophic ulcers of the lower extremities influences the total outcomes of surgical management. A conclusion was made on the necessity of a compulsory dynamic scrutiny of the microbial flora of the lower-limb atherosclerotic ulcer, which makes it possible to carefully select individual antibacterial bactericidal therapy with due regard for microbial sensitivity to bactericidal agents. PMID- 19806938 TI - [Assessment of the level of homocysteine and the state of haemostasis in patients with and without type 2 diabetes mellitus in remote terms after reconstructive operations on the aorto-iliac segment]. AB - The authors carried out a comparative analysis of the level of homocysteine and the state of haemostasis in patients with and without type 2 diabetes mellitus in the remote terms after endured reconstructive operations on the aorto-iliac segment. They examined a total of eighty-eight patients who had endured reconstructive operations on the aorto-iliac segment at various terms. Of these, forty-two patients were found to have a severe course of type 2 diabetes mellitus (59.9% with decompensation) and forty-six subjects without diabetes constituted the group of comparison. The average age of the patients amounted to 61.9 +/- 1.25 years, with all being smokers. The following parameters were assessed: patency of the bypasses and major arteries of the lower limbs (LL), homocysteine (Hey), fibrinolytic activity, fibrinogen, activated partial thromboplastin time (aPTT), factor XIII, thrombin time, prothrombin index, activity of antithrombin III (AIII), platelet aggregation with ADP, and glycosylated haemoglobin (Hb Aic). PMID- 19806939 TI - [Methods of correcting hepatic arterial blood supply for subsequent implantation of the "port-catheter" infusion system]. AB - The study was undertaken to investigate the feasibility of using surgical and roentgenoendovascular redistribution of the hepatic arterial blood flow for adequate implantation of the "port-catheter" infusion system. Between December 2001 and March 2008, we performed surgical (n = 25) or transcutaneous (n = 22) implantations of the infusion systems for carrying out regional chemotherapy in a total of forty-seven patients presenting with hepatic metastases of colorectal carcinoma. Anatomical variants of the hepatic arteries were observed in eleven cases. The blood stream was corrected by means of either transcatheter embolization (n = 7), ligation (n = 2), or transposition (n = 2) of the aberrant arteries. No complications were encountered. In all the cases, the "port catheter" system was implanted successfully, with adequate hepatic perfusion achieved. Both transcatheter embolization and surgical reconstruction turned out to be efficient methods in correction of the blood flow in various anatomical structures of the hepatic arteries, thus favourably contributing to increased efficacy of regional chemotherapy. PMID- 19806940 TI - [Portacaval anastomoses in the basin of the left internal spermatic vein in patients with varicocele]. AB - The article deals with the findings obtained in a roentgenological examination of the left renal vein and left internal spermatic vein in a total of one hundred and eighty-three 16-to-46-year-old patients presenting with varicocele. Analysing the roentgenograms revealed that four patients had portacaval anastomoses in the basin of the left internal spermatic vein (ISV), with the blood flow being directed from the system of the portal vein into the system of the venae cavae. Retrograde contrasting of the inferior mesenteric veins showed a sharp deceleration of the contrast medium to the portal vein with an apparent obstacle to the blood flow in the projection of the intersection of the inferior mesenteric vein, lower contour of the pancreas and the aorta. In two cases, the "mesentericospermatic" venous shunt into the system of the left ISV was the only visible cause of varicocele. PMID- 19806941 TI - [Minimally invasive interventions in surgical management of varicose diseases and its relapses]. AB - Over the past several decades, many surgical methods of treatment for varicose disease have been implemented into clinical practice. Nonetheless, an intrinsically too traumatic nature thereof unsatisfactory cosmetic outcomes obtained, long-term postoperative rehabilitation required, and high recurrence rates reported send us in search of minimally invasive but at the same time highly efficient methods of treatment for varicose disease. The present article deals with the experience gained by Russian and foreign authors in and suggesting efficiency of novel minimally invasive technologies used for treatment of both newly diagnosed varicose disease and its relapsing forms. This is followed by analysing different variants of the procedural techniques involved. PMID- 19806942 TI - [Radiofrequency ablation of the great saphenous vein in lower-limb varicose disease]. AB - The authors herein share their experience gained in comprehensive treatment of patients diagnosed with lower-limb varicose disease treated by means of one the most state-of-the-art technologies, i. e., radiofrequency ablation. The work deals with the outcomes of managing a total of seventy-eight patients suffering from lower-limb varicose diseases with valvular insufficiency of the great saphenous vein. All the patients were subjected to radiofrequency ablation of the compromised great saphenous vein by means of the portable radiofrequency device "Surgitron" manufactured by the Company "Ellman International" (USA). Examination included ultrasonographic duplex and triplex scanning of the veins of the lower extremities. Intraoperative radiofrequency ablation of the great saphenous vein was performed by means of an original appliance consisting of a metallic probe whose length corresponded to that of the lower extremity concerned, with a figure of-eight olive at the distal end and external plastic tube catheter. The construction of the probe made it possible to maximally free the great saphenous vein from blood through the lateral ports of the catheter, to reduce the risk of complications, to diminish the duration of the intervention, and to increase its effectiveness. Efficacy of radiofrequency ablation of the great saphenous vein amounted to 93.6%, with recanalization of the greater saphenous vein being established in five (6.4%) patients. Good functional and cosmetically pleasing therapeutic outcomes were observed in the majority of the patients (87.2%). PMID- 19806943 TI - [Sequence of venous blood flow alterations in patients after recently endured acute thrombosis of lower-limb deep veins based on the findings of ultrasonographic duplex scanning]. AB - This study was undertaken to investigate the sequence of alterations in the venous blood flow to have occurred within the time frame of one year after sustained acute thrombosis of the lower-limb deep veins, which was carried out using the standard technique of ultrasonographic duplex scanning. A total of thirty-two 24-to-62-year-old patients presenting with newly onset acute phlebothrombosis were followed up. All the patients were sequentially examined at 2 days, 3 weeks, 3 months, 6 months and 12 months after the manifestation of the initial clinical signs of the disease. Amongst the parameters to determine were the patency of the deep veins and the condition of the valvular apparatus of the deep, superficial and communicant veins. According to the obtained findings, it was as early as at the first stage of the phlebohaemodynamic alterations after the endured thrombosis, i. e., during the acute period of the disease, that seven (21.9%) patients were found to have developed valvular insufficiency of the communicant veins of the cms, manifesting itself in the formation of a horizontal veno-venous reflux, and 6 months later, these events were observed to have occurred in all the patients examined (100%). Afterwards, the second stage of the phlebohaemodynamic alterations was, simultaneously with the process of recanalization of the thrombotic masses in the deep veins, specifically characterized by the formation of valvular insufficiency of the latter, manifesting itself in the form of the development of a deep vertical veno-venous reflux, which was revealed at month six after the onset of the disease in 56.3% of the examined subjects, to be then observed after 12 months in 93.8% of the patients involved. Recanalization of thrombotic masses was noted to commence 3 months after the onset of thrombosis in twelve (37.5%) patients, and after 12 months it was seen to ensue in all the patients (100%), eventually ending in complete restoration of the patency of the affected veins (to have occurred in 25% of the cases). Of special interest was the finding that insufficiency of the ostial valve of the great saphenous vein, manifesting itself by a superficial vertical veno-venous reflux, was revealed only in two (6.25%) patients examined 12 months after the onset of the disease, which may be regarded as the third stage of the phlebohaemodynamic alterations. That low prevalence and no evidence of varicose transformation of superficial veins appear to suggest an important part they play in compensation of the venous outflow from the extremity affected. PMID- 19806944 TI - [Efficacy of endovasal laser coagulation depending on the dose of laser energy]. AB - Presented herein is the authors' experience with surgical treatment of 85 lower extremities by means of endovasal laser coagulation. Shown are the energy-related characteristics of coagulation, followed by analysing the causes of unsatisfactory poor outcomes therapeutic as exemplified on their own experience. Proposed is an adequate selection of energy-related parameters of laser radiation in order to obtain stable occlusion. Given are the literature data on using the method concerned, with a detailed discussion of various points of view on the problem concerning selection of an efficient dose of the laser energy in order to attain adequate occlusion. PMID- 19806945 TI - [Surgical treatment of the patients with fibromuscular dysplasia of internal carotid arteries]. AB - The results of surgical treatment of 78 patients with fibromuscular dysplasia (FMD) of internal carotid artery (ICA) were described in the article. FMD ICA was combined with atherosclerosis of carotid artery at 68 (87,2%) patients and with pathological kinking of ICA at 64 (82%) patients. We provide information about frequency of systemic inflammation response syndrome at FMD ICA, and necessity of its treatment for improvement of surgical outcomes in these patients. The access to extracranial portion of ICA was shown which is necessary for an adequate estimation of changes artery at fibromuscular dysplasia. The role of spastic component in development of characteristic changes at FMD ICA was proved. We propose the specific approach for choosing the method of reconstruction of ICA and definition of the indications to prosthetision at FMD. PMID- 19806946 TI - [Indications for surgical management of patients with haemodynamically insignificant stenoses of carotid arteries]. AB - We examined a total of 56 patients presenting with haemodynamically insignificant stenoses of carotid arteries, with a narrowing of the internal carotid artery by an atherosclerotic plaque varying from 30% to 55%, to have also analysed the relationship between the sonographic and morphological peculiarities of atherosclerotic plaques of the carotid arteries and severity of cerebrovascular insufficiency. Of these, 27 patients were subjected to carotid endarterectomy (CEE). The long-term outcomes were studied during the follow-up period lasting up to 5 years. The incidence rate of neurological deficits turned out to be lower in the postoperative-group patients as compared with those from the control group. A conclusion was made that both the carotid artery stenosis degree and the structure of an atherosclerotic plaque appear to be equally important criteria in assessing the indications for surgical management. PMID- 19806947 TI - [Mini-approach in surgery of carotid arteries]. AB - The purpose of this study was to assess the outcomes of using a mini-approach in surgery of carotid arteries. From September 2006 to December 2007, one hundred and twenty-nine patients (with a total of 139 operations sustained) underwent interventions on the carotid bifurcation for atherosclerotic lesions and pathological tortuosity of the carotid arteries. The patients were subdivided into two groups. Group A consisted of those operated on from the standard access to the carotid arteries, with Group B comprising the patients undergoing interventions on the carotid bifurcation from a mini-approach (a cutaneous incision less than 5 cm long, averaging 4.7 +/- 0.3 cm). The control Group C was composed of forty-two patients having endured operations on the carotid arteries from the conventional access performed at other specialised departments. In the patient groups referred to above, we compared the incidence rate of perioperative complications (stroke/death), lesions of the cranial and cervical nerves, as well as wound complications. In Group A patients, there were two patients with stroke, one of whom had eventually died. Neuropathies of the cranial and cervical nerves in this group of patients were observed to have developed in eight (6.2%) and ninety-two (71.3%) cases, respectively, and in Group C patients--in eleven (28.0%) cases. Cervical haematoma was observed in fourteen (10.9%) subjects. The average duration of the postoperative hospital stay amounted to 6.8 +/- 1.1 days. Amongst the Group B patients, cervical haematoma was noted in two (3.2%, P<0.0001) cases only. A lesion of the cranial nerves (that of the superior laryngeal nerve) took place in two patients (P<0.0001), and that of the cervical nerves--in four patients (P<0.0001). No cheloid cicatrices were observed. The patients reported excellent and aesthetically pleasing surgical outcomes. The average duration of the postoperative hospital stay amounted to 2.8 +/- 0.69 days (P = 0.002). Hence, operations on the carotid bifurcation may be successfully performed from a mini-approach, with thus discomfort kept at a minimum and rapid rehabilitation of patients. PMID- 19806949 TI - [Nonreconstructive revascularizing operations in limb ischaemia]. AB - Presented herein are the findings of experimental studies substantiating interrelationship between osteogenesis and blood circulation, as well as effects of surgical interventions based on excitation of reparative regeneration of the bone and aimed at improvement of blood circulation in the extremities. Reflected also are the main indications for the use of each operative intervention concerned. Based on the clinical findings obtained by follow up of 1,477 patients operated on for limb ischaemia secondary to obliterating arterial lesions efficacy of the methods of treatment devised is demonstrated. PMID- 19806948 TI - [Experience with vascular biografts "KemAngioprotez" in reconstructive surgery of lower-limb major arteries]. AB - Presented herein is the experience with the Russian-made vascular biografts "KemAngioprotez" in 149 patients suffering from atherosclerotic occlusions of the major arteries of the lower extremities and in one female patient with an injury to the superficial femoral artery. A total of 150 reconstructive operations were performed on all segments of the major arterial bed of the lower limbs. The early postoperative period was accompanied by 14 thromboses of the vascular biograft after femoropopliteal reconstruction. It was determined that the likelihood of thrombosis of the vascular biograft depended upon the type of the distal anastomosis, increasing in establishing an "end-to-side" distal anastomosis with the popliteal artery as compared with the "end-to-end" anastomosis technique. No cases of either aneurysmatic deformity, rupture of the vascular graft nor its infection in the early postoperative period were observed. The vascular biograft proved convenient and may therefore be successfully used to replace synthetic vascular grafts and autovenous transplants in reconstruction of the external iliac, femoral and popliteal arteries. PMID- 19806950 TI - [Surgical management of occlusion of the celiac trunk and an aneurysm of the inferior pancreatoduodenal artery]. AB - Presented in the article is a clinical case report of successful management of an aneurysm of the inferior pancreatoduodenal artery, having resulted from occlusion of the celiac trunk and a compensatory increase in the blood flow along it from the superior mesenteric artery into the basin of the celiac trunk. In the case report described, the authors used the technique of open surgical revascularization of the celiac trunk and autovenous prosthetic repair of the aneurysmatically altered inferior pancreatoduodenal artery. The surgical decision making was determined by the fact that the commonly accepted endovascular by-pass of the aneurysm would have resulted in the development of ischaemia in the basin of the celiac trunk and unpredictable alterations in the organs supplied thereby. PMID- 19806951 TI - [A case report of successful surgical management of lower-limb critical ischaemia on the background of femoropopliteal atherosclerotic aneurysms]. AB - Presented herein is a case report of successful surgical management of a male patient presenting with lower-limb critical ischaemia on the background of thrombosis of femoropopliteal atherosclerotic aneurysms and the presence of a necrobiotic process on the stump of the foot. The reconstructive intervention performed resulted in salvation of the extremity, followed by uneventful wound healing on his left foot with complete medical and social rehabilitation of the patient. PMID- 19806952 TI - [Statins in practice of the vascular surgeon and cardiologist]. AB - The article deals with a review of the present-day literature data regarding administration of lipid-decreasing agents--statins in clinical practice of cardiologists and heart surgeons. In the context of the constantly accumulating evidence-based findings concerning efficacy of administration of statins in patients suffering from cardiovascular diseases, the authors describe the importance of therapy with statins in order to prevent complications after atherothrombosis. PMID- 19806953 TI - [Present-day possibilities of correcting impaired autoregulation of the cerebral circulation]. AB - Reviewed herein is the present-day literature data concerning the causes leading to the development of ischemic stroke. Special attention is drawn to the haemodynamic mechanism, of cerebral circulation impairment. Also discussed is the fed of the onset of such disorders in patients suffering from peripheral atherosclerosis during reconstructive surgical vascular interventions, in relation with a multifocal nature of the lesion. Presented are modern findings on diagnostic methods making it possible to assess autoregulatory disorders of cerebral circulation, the main causes of these impairments, as well as potential principles of correction thereof. PMID- 19806954 TI - [Surgical management of a patient presenting with stage IV lower limb ischaemia and being on programmed haemodialysis]. AB - Presented herein is a clinical case report of a male patient suffering from ischaemic gangrene of the foot on the background of an atherosclerotic-genesis multifocal occlusive-and-stenotic lesion of the major arteries of the lower extremities and undergoing programmed haemodialysis for terminal-stage chronic renal insufficiency. The patient was successively subjected to three-stage surgical management, with the first stage consisting of revascularization of the extremity involved by means of "in-situ" femoropopliteal auotovenous bypass grafting, the second comprised resection of the calcaneum with the removal of the osteomyelitis focus, and the third one was composed of dermatoautoplasty of the defect of the foot with a split cutaneous flap. PMID- 19806955 TI - Perceptual high-lights. PMID- 19806956 TI - Single units and sensation: a neuron doctrine for perceptual psychology? PMID- 19806957 TI - From maverick to mainstream. PMID- 19806958 TI - Single units and sensation: still just as relevant today. PMID- 19806959 TI - Translational and reflectional priming invariance: a retrospective. AB - Biederman and Cooper (1991a) showed that the presentation of a briefly presented image of an object at one position in the visual field facilitated its identification, as assessed by naming speed and accuracy, several minutes later. The facilitation was unaffected by a translation or a reflection of the stimulus. A component of this priming was visual rather than basic-level conceptual or lexical in that there was less facilitation for an object with the same name (and basic-level class) but a different shape. The invariance of priming to view variables has stood up well over the years and appears to be a general phenomenon -as long as the original structural description can be readily resolved--in that it has also been observed for variations in size and orientation in depth. Although priming was unaffected by a change in position, we documented that there was explicit memory for the position (and orientation and size) of the stimulus. The existence of two forms of representation from the identical stimulus presentation--one invariant and the other dependent on view variables--poses a challenge as to what can be concluded about view invariance from single-unit activity. PMID- 19806960 TI - Challenges in object recognition: selectivity vs invariance. PMID- 19806961 TI - Challenges in object recognition: selectivity vs invariance. PMID- 19806962 TI - Invariant parts of a citation classic. PMID- 19806963 TI - Vision with isoluminant colour contrast: 1. A projection technique and observations. PMID- 19806964 TI - All's jazzy and unstable on the colour front: the impact of Gregory's pioneering paper on vision at equiluminance. PMID- 19806965 TI - Unlocked edges or contradictory edges? Is there a master signal for edge locking? PMID- 19806966 TI - Colour function. PMID- 19806967 TI - General Tau Theory: evolution to date. PMID- 19806968 TI - The (current) future is here! PMID- 19806969 TI - Lee's tau operator. PMID- 19806970 TI - Lee's tau operator. PMID- 19806971 TI - Lee's tau operator. PMID- 19806972 TI - Surfaces revisited. PMID- 19806973 TI - Revisiting the relationship between transparency, subjective contours, luminance, and colour spreading. PMID- 19806974 TI - Clear waters, murky waters: why transparency perception is good for you and underconstrained. PMID- 19806975 TI - From neon colour spreading to stratification and mechanisms. PMID- 19806976 TI - Looking back on PMF. PMID- 19806977 TI - PMF: climbing the disparity gradient for twenty years. PMID- 19806978 TI - PMF and its influence on computational stereo. PMID- 19806979 TI - Computing stereoscopic depth in natural images. PMID- 19806980 TI - "Those were the days, my friend, we thought they'd never end...". PMID- 19806981 TI - Eye movements and landing positions in reading: a retrospective. PMID- 19806982 TI - Rayner's 1979 paper: a brief summary and evaluation. PMID- 19806983 TI - Preferred viewing locations: a validation and an extension. PMID- 19806984 TI - Fixation locations within words. PMID- 19806985 TI - Motion parallax as an independent cue for depth perception: a retrospective. PMID- 19806986 TI - Motion parallax with and without active head movements. PMID- 19806987 TI - Motion parallax with and without active head movements. PMID- 19806988 TI - What Rogers and Graham showed us about how vision works. PMID- 19806989 TI - The paradoxes of parallaxes. PMID- 19806990 TI - The Thatcher illusion 28 years on.... PMID- 19806991 TI - Mrs Thatcher and the bikini illusion. AB - In 1980, Pete Thompson published his Mrs Thatcher illusion, one of the most astonishing and unexpected illusions of the century. It has been deservedly cited hundreds of times. The Mrs Thatcher illusion, together with upside-down smiles and upside-down bodies, can reveal selective breakdowns of configural versus featural processes in face perception. PMID- 19806992 TI - The Thatcher illusion: now you see it, now you don't. PMID- 19806993 TI - The enduring nature of the Thatcher illusion. PMID- 19806994 TI - 'Two categorical stages of object recognition': a retrospective. PMID- 19806995 TI - Multiple representations for perceptual categorisation. PMID- 19806996 TI - Fractionating object recognition. PMID- 19806997 TI - Mapping a model of object recognition. PMID- 19806998 TI - Long QT and Brugada syndromes: a new insight into arrhythmogenesis and sudden death. PMID- 19806999 TI - Remembering 'EO' and the 5 Es. PMID- 19807000 TI - Measuring the quality of patient care goes beyond just numbers. PMID- 19807001 TI - Health care debate 101. PMID- 19807002 TI - Outside the box: Home dialysis can be a problem-solver for common in-center woes. PMID- 19807003 TI - Coalition releases position statement on protecting home dialysis in bundled ESRD payment. PMID- 19807004 TI - How to make the new Conditions for Coverage work in your dialysis clinic. PMID- 19807006 TI - Polishing up professionalism: the patient-centered professional. PMID- 19807007 TI - The transition of pediatric to adult-centered health care. PMID- 19807008 TI - Stroke: recognising the signs. PMID- 19807010 TI - Trauma centres: a capital idea. PMID- 19807011 TI - Road Safety Week. PMID- 19807012 TI - Emergency care for people with dementia. AB - As part of its commitment to improving dementia services, the Scottish Government has published six recommendations for developing care in emergency departments. This article looks at how each of these recommendations can be implemented by emergency nurses to make departments safer places for older people with confusion. PMID- 19807013 TI - Pandemic flu: lessons from the Toronto SARS outbreak. AB - A review of the severe acute respiratory syndrome outbreak in Ontario in 2003 found that there had been several faults in healthcare services and that these had contributed to the spread of infection and the deaths of two nurses. This article argues that, six years on, these faults have not been rectified and that Ontario healthcare services may be unprepared for an H1N1 outbreak. The assistant deputy public health minister of Toronto offers her response. PMID- 19807014 TI - Casting: below-elbow back slabs. AB - This article continues a short series of articles on how to apply casts. It examines how below-elbow back slabs should be applied following fracture of the distal forearm and wrist. PMID- 19807015 TI - Accurate diagnosis of right ventricular infarction. AB - Effective treatment for acute myocardial infarction (AMI) is dependent on accurate' diagnoses of the site of infarction. Left ventricular myocardial infarction (LVMI) is the most common form of infarction, of which inferior is the most common site. Patients who have experienced inferior infarctions have a better survival rate than those who have experienced AMIs at other sites. Around one quarter of AMIs affect the right ventricle, and the risk of morbidity for patients who have experienced right ventricular myocardial infarctions is greater than that for patients who have experienced LVMIs. Some authors suggest this is because right-sided heart dysfunction is rarely assessed in standard electrocardiography (ECG), and so opportunities for accurate diagnoses are missed. This article describes how non-standard precordial ECG lead placement can increase the likelihood of diagnosis and therefore improve patient outcomes. PMID- 19807016 TI - Acute stroke management in emergency departments. AB - The introduction of thrombolysis and dedicated stroke units have greatly improved the management of people who have had a stroke. However, for the benefits of these advances to be fully realised, emergency department nurses must be able to recognise the signs of stroke and act appropriately and quickly. With thrombolysis, timing is all-important, so practitioners need the knowledge and skills to assess individuals' suitability for this treatment, request immediate brain scans and refer patients directly to stroke units. PMID- 19807017 TI - New horizons. PMID- 19807018 TI - [The concept of transmembrane asymmetry of lateral domains in biomemranes and influenza virus envelope fine structure]. AB - The molecular architectures of enveloped viruses are one demonstrative example of perfectly arranged macromolecular complex that is achieved through the structural specificity of virus assembly. Virus morphogenesis is a multi-step process that depends on the concerted actions of many viral and cellular components as well as fitted organization of main viral constituents. Viral envelope was shown to be composed of the mixture of lipid raft and non-raft domains. The domains are recruited from the host-cell membrane as discrete well-ordered lipid-protein units in the process of virus assembly. Raft-like nature of influenza virus A envelope was visualized using a novel approach of cold solubilization of the detergent-resistant membranes from intact influenza virus A virions with the mixture of two non-ionic detergents drastically differing in their raft solubilizing activities, NP40 and octyl-glucopyranoside. In the view of this methodological approach, the virus envelope is apparently an ensemble of platforms which are flexibly joint in the viral envelope, and composed of surface glycoproteins (hemagglutinin and neuraminidase), matrix M1 protein and lipids. The modern concept of transmembrane asymmetry of lateral domains in biological membranes was involved to explain the solubilization mechanism revealed. Using principles of this concept we suggest matrix M1 protein shell as a structure forming base to support asymmetrical rafts in the virus envelope. PMID- 19807019 TI - [Detection of LY6G6D gene exon-intron structure,]. AB - Earlier we have shown high frequency of loss of heterozygosity of microsatellite marker D6S273 within HLA III class region in DNA samples from cervical intraepithelial neoplasias and cervical cancer. According to publications three genes were identified in this region. For detection of D6S273 position we used in silico analysis of mRNA sequences deposited in GenBank (NCBI) and investigated LY6G6D gene expression in tumor cell lines. LY6G6D gene exon borders were analyzed with 5'- or 3'-rapid amplification of cDNA ends. We have found that LY6G6D gene consists of 9 exons and includes two earlier identified genes G6D and G6F. Microsatellite D6S273 is located in the last 8 intron of LY6G6D gene. The third gene LY6G6E consisting of four exons is located in 6 intron of LY6G6D gene in the opposite orientation. We suggest that LY6G6D gene is coding three main mRNA transcripts in the same open reading frame but differ in exon composition: MEGT1 consists of 1-4, 8, 9 exons, G6F consists of 1-6 exons and G6D consists of 7-9 exons of LY6G6D gene. High homology with immunoglobulin superfamily within 20 120 aminoacids of MEGT1 and G6F proteins is shown by in silico translation of their mRNAs. PMID- 19807020 TI - [Analysis of 5'-leader regions in protozoa type Apicomplexa and red algae plastids]. AB - Apicomplexan parasites contain the apicoplasts, a chloroplast-like organelle. By means of the multiple alignments of 5'-leader regions for the plastid-encoded genes, we found some conserved non-coding regions in the parasites as well as in red algae. We suppose these regions are the sites for interaction between the RNA and regulatory proteins. Conservative sites was found upstream gene ycf24, required for [Fe-S]-clasters development, and upstream some other genes. In particular, in Toxoplasma gondii a simultaneous regulation is predicted for three genes ycf24, rps4 and rpoB. Our prediction corresponds to the known data that T. gondii apicoplasts in its entirety are required for a short time only, but ones procure pathogenesis. Another site is predicted upstream gene rpoB, that encodes beta-subunit of the RNA polymerase, in red algae Porphyra spp. as well in Eimeria tenella and Theileria parva. PMID- 19807021 TI - [Novel 35-bp insertion in visfatin gene in Chinese cattle]. AB - Visfatin is a protein that is preferentially produced in visceral adipose tissue. Both its tissue expression and secreted plasma levels increase in parallel with obesity. Therefore, the present study is focused on detecting insertion in bovine visfatin gene and analyzing its effect on growth traits in four Chinese cattle breeds (Nangyang, Qinchuan, Jiaxian, and Chinese Holstein) using PCR-SSCP, DNA sequencing and agarose electrophoresis methods. For the first time, a 35-bp insertion was described in the gene and two alleles were revealed: Wand I. The chi2-test analysis demonstrated that all breeds were in agreement with Hardy Weinberg equilibrium (P > 0.05). The associations of the novel 35bp-insertion of visfatin gene with growth traits of Nanyang cattle at 6, 12, 18, and 24 months old were analyzed: 6-month-old cows with genotype WW had greater birth weight than cows with genotype II and WI (P < 0.01 or P < 0.05). These results suggest that the presence of 35bp-insertion in visfatin gene may influence the birth weight and bodyweight in 6-month-old cows. PMID- 19807022 TI - [Proteomic expression analysis of human colorectal cancer: of soluble overexpressed proteins]. AB - Colon cancer is one of the leading causes of cancer deaths in developed countries due to the absence of tumor specific markers for early diagnosis of the disease, providing adequate sensitivity. Search for diagnostic markers of various types of cancer by proteomic approaches has been limited by large differences in protein centration. We used preliminary extraction of major cellular proteins by 0.2 M sodium chloride in presence of nonionic detergent NP-40 in order to raise the sensitivity of the 2D PAGE detection of low-abundant soluble proteins, some of which may penetrate in blood circulation during carcinogenesis. Application of this procedure prior to 2D comparative analysis of proteomes of normal tissues and matched colon cancer specimens led to selection of ten proteins, which are frequently overexpressed in colon adenocarcinomas. Mass-spectrometric identification of selected proteins led to discovery of two novel protein markers of colon tumors--TAF9 and CISH. Low level of CISH expression in various tissues suggests that it is a novel prospective marker for diagnosis of colon cancer. PMID- 19807023 TI - [Orthopoxvirus genes for kelch-like proteins. III. Construction of mousepox (ectromelia) virus variants with targeted gene deletions]. AB - Mousepox (ectromelia) virus genome contains four genes encoding for kelch-like proteins EVM018, EVM027, EVM150 and EVM167. A complete set of insertion plasmids was constructed to allow the production of recombinant ectromelia viruses with targeted deletions of one to four genes of kelch family both individually (single mutants) and in different combinations (double, triple and quadruple mutants). It was shown that deletion of any of the three genes EVMO18, EVM027 or EVM167 resulted in reduction of 50% lethal dose (LD50) by five and more orders in outbred white mice infected intraperitoneally. Deletion of mousepox kelch-gene EVM150 did not influence the virus virulence. Two or more kelch-genes deletion also resulted in high level of attenuation, which could evidently be due to the lack of three genes EVM167, EVM018 and/or EVM027 identified as virulence factors. The local inflammatory process on the model of intradermal injection of mouse ear pinnae (vasodilatation level, hyperemia, cutaneous edema, arterial thrombosis) was significantly more intensive for wild type virus and virulent mutant deltaEVM150 in comparison with avirulent mutant AEVM167. PMID- 19807024 TI - [Different instability of microsatellite CAG repeat in two groups of haplotypes of the human mitochondrial gamma-DNA-polymerase gene]. AB - Polymorphism of two single nucleotide loci rs758130 (T/C) and rs2238296 (T/C) of mitochondrial DNA-polymerase gamma (POLG1) gene has been studied in individuals of different ethnoracial ancestry in Russians and Buryats) with already known genotypes of microsatellite CAG-repeat of this gene. It was found that alleles with the number of repeats unequal to 10 are more often detected on the background of haplotype TT than CC. Results of phylogenetic analysis of the POLG1 gene intron 2 nucleotide sequences in humans and chimpanzee have shown that haplotype TT heterogeneous in relation to CAG-repeat polymorphism appears to be evolutionary younger than haplotype CC. The data obtained can be used for further studies of relationships between CAG-repeat polymorphisms and male infertility. PMID- 19807025 TI - [Isolation and functional analysis of cotton universal stress protein promoter in response to phytohormones and abiotic stresses]. AB - The 949 bp promoter fragment upstream from the translation initiation site of the GUSP gene encoding a universal stress protein was isolated from the genomic DNA of Gossypium arboream. Some putative cis-acting elements involved in stress responses including E-box, ABRE, DPBF-box, and MYB-core elements were found in the promoter region. In an Agrobacterium-mediated transient expression assay, strong activation of the GUSP full promoter region occurred in tobacco leaves following dehydration, abscisic acid, salt, heavy metal, gibberellic acid and dark treatments. Deletion analysis of the promoter revealed that the dehydration, abscisic acid and salt responses were affected by the deletion between -208 and 949 bp and showed 2-4-fold induction. However, in response to dark, gibberellic acid and heavy metals the induction was only 2-fold. This is an important study as no report of this universal stress protein promoter is available in literature. PMID- 19807026 TI - [Small interfering RNAs targeting human myostatin]. AB - Ten human myostatin small interfering RNAs which sequence was found by two different software products were synthesized and tested for activity. It was found that three of them have pronounced biological activity and decrease myostatin mRNA level to 22-27% of control value. These small interfering RNAs stimulate human myoblast proliferation and decrease their differentiation reliably. The obtained small interfering RNAs can be used for development of new approaches for the treatment of sarcopenia and different myodystrophies. PMID- 19807027 TI - [Adaptation of mycoplasmas to stressful factors: nucleotide sequences nonregistered in vegetative forms of M. galisepticum S6 cells are revealed in nonculturable forms of the mycoplasma]. AB - Adaptation of M. gallisepticum S6 to unfavorable environmental conditions is connected with transformation of the vegetative forms of the mycoplasma cells to the viable but non-culturable (VBNC) forms. The vegetative forms and the VBNC forms differ in the spectrum of the PCR-products that was forming due to amplification of the nucleotide sequence of the pvpA-gene coding the able cytoadhesion protein. As to vegetative forms of the mycoplasma the only amplicon, containing one open reading frame (1086 b.p.) with a high homology (97%) to the pvpA-gene of M. gallisepticum R and Pendik is detected. In the case of VBNC forms of M. gallisepticum S6, the additional amplicons besides those indicating the pvpA-gene of the mycoplasma are observed. In the nucleotide sequences of the additional amplicons, the open reading frames are detected that are not registered in the database of the complete sequence of the mycoplasma genome. A high homology (54-55%) of the nucleotide sequences of the pvpA-gene and the additional pvpA-amplicons allows to suggest that thepvpA-gene sequence seems to be a basis for forming new regions within the mycoplasma genome during adaptation of the bacterium to unfavorable environmental conditions. PMID- 19807028 TI - [Preparation and characterization of mouse embryonic fibroblasts with K72W mutation in somatic cytochrome C gene]. AB - Mouse embryonic fibroblasts (MEF) with point mutation in somatic cytochrome C gene were generated and characterized. It was shown that substitution of lysine for tryptophan in position 72 (K72W) decreased the proapoptotic functions of cytochrome C in response to staurosporin treatment without disrupting its respiratory functions. The presence of this mutation did not affect the pattern of cytochrome C gene expression or its localization inside the cell. These cell cultures therefore represent an interesting model for study apoptotic signaling and physiological functions of cytochrome C. PMID- 19807029 TI - [New putative control elements in the promoter OF CXCL13 chemokine gene, a target of alternative NF-kappaB pathway]. AB - We searched the proximal promoter region of CXCL13/BLC chemokine gene for new putative control elements, including potential NF-kappaB binding sites. Using electrophoretic mobility shift assay and reporter gene analysis we identified two new promoter elements. The first element contains Rel/NF-kappaB binding site and seems to participate in inducible gene expression, while another site binds transcription factor Sp1 and is critical for basic transcription. It is the first indication that alternative NF-kappaB pathway target genes are probably cooperatively controlled by factors Rel/NF-kappaB and Sp1. Identification of a functional Sp1 site in the promoter of a target gene of alternative NF-kappaB pathway will be useful for investigation of molecular mechanisms and signal pathwaysinvolved. PMID- 19807030 TI - [Conserved structural features of ETS domain--DNA complexes]. AB - ETS proteins comprise a family of widespread transcription factors regulating expression of many animal genes. Structurally ETS proteins are characterized by conserved DNA-binding ETS domain, recognizing DNA sequences with trinucleotide GGA. Comparative analysis of structural features of ETS domain-DNA complexes was carried out and conserved contacts, important in terms of interaction stability and specificity were identified. The analysis revealed 9 conserved hydrogen bonds with DNA backbone phosphates and 2 conserved bidentate hydrogen bonds with DNA major groove atoms, one conserved hydrophobic cluster on protein-DNA interface, important for binding site recognition, and 12 conserved water molecules, possibly mediating ETS domain-DNA interaction. The results are represented in specialized data bank of protein-DNA complexes NPIDB. PMID- 19807031 TI - [The interaction of dimeric form of RNA retroviruses with paromomycin and magnesium: using the fluorescence of 2-aminopurine]. AB - The studying of the dimeric RNA structural organization is a step in the understanding of retroviruses genomic RNA dimerization. The RNA kissing loop dimer rearrangement into the extended dimer occurs during the maturation of virus particle. The inhibition of the extended dimerformation can be caused by ligands interaction with RNA kissing loop dimer. Here, we study the interaction of the dimeric RNA with paromomycin and magnesium ions. RNA dimers were formed from the different hairpin RNA having the complementary sequences in the loop. To detect the structural features of RNA dimers and influence of the ligands we used the 2 aminopurine fluorescence (2-AP) incorporated in one of two RNA hairpin sequences. It was observed that the 2-AP fluorescence increases under dimer RNA interaction with paromomycin. The growing of 2-AP fluorescence can be explaining by fluorescent base flipping out from the RNA structure. The binding affinity and stoichiometry to the kissing loop a nd extended dimers were found by2-AP fluorescence alteration. It turned out, that one RNA dimer binds with two paromomycin molecules; the binding constants for both dimers type were approximately the same and about 3 x 10(5) M(-1). The competition of antibiotic and Mg2+ ions binding were revealed. It was found that one paromomycin molecules displaced by one Mg2+ ion. PMID- 19807032 TI - [Selection of DNA aptamers, specifically interacting with fibrillar form of the yeast Sup35 protein]. AB - Single-stranded DNA aptamers interacting with fibers formed by the Sup35 protein of Saccharomyces cerevisiae were obtained by the SELEX procedure. Specificity of interaction with Sup35p is investigated for 10 from total of 40 selected aptamers. It is shown that 9 aptamers bind to fibrillar Sup35p and not to monomeric form of the protein. The rate of dissociation constant of aptamer-fiber complex varies from 0.1 to 1 microM for different aptamers. Selected aptamers can be used to study prionization of Sup35. PMID- 19807033 TI - [X-ray interferometry of the axial movement of myosin heads during muscle force generation initiated by T-jump]. AB - The interference fine structure of the M3 reflection on the low-angle x-ray diffraction patterns of muscle fibres is used for the measurements of axial movements of myosin heads with a precision of 0.1-0.2 nm. We have measured changes in the M3 interference profile during tension rise induced by a 5 to 30 degrees C temperature jump in thin bundles of contracting fibers from rabbit skeletal muscle. Interpreting the data with a point diffractor model gives an estimate for the axial movement of the myosin heads during force rise of less than 0.6 nm. Modifications of the point diffractor model are discussed. We show that our experimental data can be explained by a model where myosin heads bind actin in a number of structurally different states. PMID- 19807034 TI - [Mutual effect of human ribosomal proteins S5 and S16 on their binding with 18S rRNA fragment 1203-1236/1521-1698]. AB - Human ribosomal proteins S5 and S16 are homologues of prokaryotic ribosomal proteins S7p and S9p, respectively, that according to X-ray crystallography data on the Thermus thermophilus 30S ribosomal subunit contact the 3'-terminal 16S rRNA region formed by helices H28-H30 and H38-H43. In the present work we report studying mutual effect of human ribosomal proteins S5 and S16 on their binding with RNA transcript corresponding to the region 1203-1236/1521-1698 of the 18S rRNA (helices H28-30 and H41-43), which is homologous to thel6S rRNA region known to contain binding site of S7p and part of binding site of S9p. It was shown that simultaneous binding of ribosomal proteins S5 and S16 with this RNA transcript causes conformational changes in it stabilizing the complex by involvement of new parts of the RNA that interact with neither S5 nor S16 in the respective binary complexes. PMID- 19807035 TI - [New program PSI protein classifier automatizes the PSI-BLAST results analysis]. AB - PSI Protein Classifier is a new program that allows to summarize the consecutive and independent PSI-BLAST iteration results. The technical opportunities of the program are explained. Two examples of the PSI Protein Classifier application are given. Iterative screening of the protein database allowed to reveal feasible evolutionary relationship among GH5, GH13, GH27, GH31, GH36, GH66, GH101, and GH114 families of glycoside hydrolases. Family GH31 is divided into 38 subfamilies on the basis of statistically significant sequence similarity analysis (E-value analysis). PMID- 19807036 TI - [Protein structure prediction from analogy. I. New database of spatially similar and dissimilar structures of protein domains for testing and optimization of prediction methods]. AB - The paper describes creation and analysis of a database 3Dfold_test. This database consists of a large set of pairs of spatially-similar structures of protein domains and of an accompanying, much larger control set of "decoys", i.e., spatially-dissimilar protein structures, having approximately the same size and compactness as each member of a pair of spatially-similar proteins. The database 3Dfold_test can be found at the site http://phys.protres.ru/resources/prediction analogy/3Dfold/. PMID- 19807037 TI - [Protein structure prediction from analogy. II. Testing of substitution matrices and pseudo-potentials used to align protein sequences with folds]. AB - The paper describes results of testing of "recognition ability" of different substitution matrices and various pseudo-potentials (taken from literature or designed by us) to recognize protein structures and produce the sequence-to structure alignments. Numerical estimates of "identifying power" of various matrices and pseudo-potentials are obtained for different levels of similarity between amino acid sequences of spatially-similar proteins. It is shown that substitution matrices work much better than pseudo-potentials at a high level of similarity of sequences of spatially-similar proteins, but some pseudo-potentials overcome substitution matrices at a low level of sequence similarity. PMID- 19807038 TI - [Cowpea mosaic virus chimeric particles bearing ectodomain of matrix protein 2 (M2E) of influenza A virus: production and characteristics]. AB - The epitope presentation system for ectodomain of M2-protein of influenza A virus (M2e) based on Cowpea Mosaic Virus (CPMV) was constructed for expression in plants Vigna unguiculata. CPMV is widely used as a vector for production of immunogenic chimeric virus particles (CVPs) bearing epitopes of different infectious human and animal pathogens. To produce chimeric CPMV virus particles in plants, two binary vectors were constructed bearing modified gene coding for S coat protein of CPMV with insertions of M2e epitopes of human influenza and bird influenza viruses. Antigenic and immunogenic properties of CVPs obtained were investigated in mice immunization experiments and it was shown that they can induce anti-M2e IgG production and partial protection mice against challenge with low doses of flu virus. However, low infectivity and immunogenicity of CPMV chimeric particles indicate the need for further optimization of plant virus based systems for M2e-epitopes presentation to use plants as a possible source of flu vaccines. PMID- 19807039 TI - [Difficilina cerebratuli gen. et sp. n. (Eugregarinida: Lecudinidae)--a new gregarine species from the nemertean Cerebratulus barentsi (Nemertini: Cerebratulidae)]. AB - A new species of aseptate gregarine, Difficilina cerebratuli gen. et sp. n. (order Eugregarinida Leger, 1900; suborder Aseptata Chakravarty, 1960; family Lecudinidae Kamm, 1922) from the gut of the White Sea nemertean Cerebratulus barentsi Burger, 1895, has been described. The electron and light microscopic data on trophozoites are presented. Their general morphology resembles the representatives of the genus Lecudina, but the features of the epicyte ultrastructure are different from Lecudina and similar to those of the Lankesteria spp. Taxonomy of the described species is discussed. Certain ultrastructural characters are included in its generic and specific diagnoses. Genus Difficilina gen. n. Type species: Difficilina cerebratuli sp. n. DIAGNOSIS: Characters of the family. Free trophozoites elongated, anterior end rounded, without hooks or exfoliations, not separated from the rest of the body, with well developed terminal smooth area. The epicytic folds undulating vertically, in cross sections--monomorphic, finger-shaped, with strongly developed cell-coat, with additional electron-dense axial structure ("middle axis") at the tops; number of rippled dense structurtes and apical filaments 3, the furthers are thick and slightly flattened in diameter. Other stages unknown. In testinal parasites of nemerteans. DIFFERENTIAL DIAGNOSIS: The new genus differs from Lecudina by presence of smooth area at the apical pole of the body and the epicyte structure: vertically undulating monomorphic finger-shaped (in cross section) epicytic folds, oligomerization of the rippled dense structures and apical filaments, and development of the "middle axis". It also differs from Lankesteria by the shape of the body, vertical undulation of the folds, and non tunicate host. Difficilina cerebratuli sp. n. DIAGNOSIS: Characters of the genus. Free trophozoites slightly bent, up to 250 x 70 microm. Anterior end with less granular cytoplasm; with feebly marked apical papilla encircled by the smooth area. Posterior end pointed. The average height of the epicytic folds is 280 nm, thickness--160 nm, average distance between folds is 130 nm. Endoplasm contains a number of accumulations of the fibrillar matter. Nucleus spherical 14-26 microm in diameter, localized in the middle or in the fore third of the body, carries 1 2 karyosomes of various size. Other stages unknown. TYPE SERIES: Microscopical slide with 7 trophozoites (Bouin, Delafield's haematoxylin) is kept in the Zoological museum of the Moscow State University (collection number: Z-2). In addition, two syntypes are observed with TEM and two other syntypes--with SEM. TYPE HOST: Cerebratulus barentsi Burger, 1895 (Nemertini: Cerebratulidae). LOCATION: Intestine (?). TYPE LOCALITY: White Sea Biological Station of the Moscow State University, Yeremeyevsky Rapid, Velikaya Salma Strait, Kandalaksha Bay, White Sea. PMID- 19807040 TI - [Description of the maritae and determination of the species status of Microphallus pseudopygmaeus sp. nov. (Trematoda: Microphallidae)]. AB - Microphallus pseudopygmaeus belongs to the "pygmaeus" microphallids, a group of closely related species with homotypic two-host life cycle. This cycle involves one intermediate host (littoral or sublittoral gastropods, mostly of the genus Littorina), and bird-invading metacercariae develop within daughter sporocysts. In spite of the fact that the name Microphallus pseudopygmaeus is widespread in special literature, the original description of this species (Galaktionov, 1980) was performed in contravention of the International Code of Zoological Nomenclature. This article is aimed to correct the above defect. The maritae of Microphallus pseudopygmaeus were obtained from the common eider ducklings which were infected experimentally with metacercariae of Microphallus sp. I Galaktionov, 1980. These metacercariae were extracted from the naturally infected mollusks Littorina saxatilis collected at the Barents Sea coast. The valid description of the species M. pseudopygmaeus, its comparison with other representatives of the "pygmaeus" microphallids and synonymy are provided. Detailed analysis of digeneans from marine and coastal birds preserved in the collections of the Zoological Institute RAS (St. Petersburg, Russia) and the Natural History Museum (London, UK) showed that in some cases M. pseudopygmaeus was misidentified as M. pygmaeus. It has been shown that M. pseudopygmaeus has an amphiboreal-arctic distribution and the spectrum of its first intermediate hosts is unequally wide for digeneans. It includes 15 species of north-Atlantic and north-Pacific mollusks belonging to different families and even orders of Prosobranchia. At the same time, the range of final hosts of M. pseudopygmaeus is limited by benthophagous marine ducks, first of all the common eider. PMID- 19807041 TI - [Severianoia blapticola sp. n. (Oxyurida: Thelastomatidae): a new nematode species from blaberid cockroaches]. AB - A new nematode species of the genus Severianoia (Schwenk, 1926) Travassos, 1929, S. blapticola sp. n., is described from the hind gut of cockroach Blaptica dubia (Serville, 1839) from laboratory culture. The main morphological feature of S. blapticola sp. n. is the presence of columnar cuticular structures of different height on the ventral side of male posterior end. In the gut of B. dubia these nematodes coexist with another thelastomatid genus, Cranifera Kloss, 1960 [C. cranifera (Chitwood, 1932) Kloss, 1960]. Males of C. cranifera are characterized by the presence of cloacal projection and columnar structures on both dorsal and ventral sides of male posterior end. PMID- 19807042 TI - [Polymorphism and intraspecific variability in the cestode Proteocephalus tetrastomus (Rudolphi, 1810) (Cestoda: Proteocephalidae), a specific parasite of smelt (Osmeridae)]. AB - Morphological variability in the cestode Proteocephalus tetrastomus parasitizing smelt in Lake Ladoga (Baltic Sea basin) and Lake Goluboye (Pechora River basin) was studied. Two forms of P. tetrastomus differing in the proglottid structure, and two variations of the ovary were discriminated. Slightly craspedote individuals with a dumbbell-like ovary were included in the first group. The second group includes distinctly craspedote individuals. It is heterogeneous by the ovary shape and comprises cestodes with two variants of the latter, dumbbell like and triangular. Both forms are present in the Lake Ladoga. The slightly craspedote form dominates, whereas the distinctly craspedote form is rare. In Lake Goluboye, P. tetrastomus is represented by the only form, distinctly craspedote with a triangular ovary. Differences in the morphometric parameters of the forms, as well as geographic and habitat distinctions, were determined. As compared to other studied cestode species of the genus Proteocephalus (P. torulosus, P. longicollis, P. percae), the smelt parasite P. tetrastomus has fewer polymorphic traits and their variations, and a much more restricted range of variability in the measurable, counting and relative characters. Analysis of literature (Willemse, 1969; Scholz and Hanzelova, 1998; Scholz et al., 2004) and original data suggest that the species is host-specific and relatively stable. It can be concluded, that P. tetrastomus is a polymorphic, complex-structured species. PMID- 19807043 TI - [Fauna of myxosporidians (Myxozoa: Myxosporea) in the scorpionfish genus sebastes from North Atlantic]. AB - Data on species composition, hosts, localities, and occurrence of myxosporidean in four host species of the scorpionfish genus Sebastes from different areas of North Atlantic and Barents Sea are presented. Thirteen myxosporidean species are recorded. PMID- 19807044 TI - [Phenomenon of Yersinia pestis biofilm formation in flea organism]. AB - For the first time substantiated was the role of the phenomenon of Yersinia pestis biofilm--extracellular matrix envelope (EME)--formation as the basis that determines the nature of plague agent interaction with flea organism. Implication of vector's proventriculus in the process of biofilm formation was demonstrated. Ultrastucture of plague microbe conglomerates in flea proventriculus and midgut was analysed and uniform mechanism of their formation was elucidated. The role of Yersinia pestis biofilm in plague agent preservation in the gut of ectoparasites and soil of rodents burrows was discussed. PCR analysis confirmed the presence of plague microbe in plague infected corpses and fleas excrements stored at +8-10 degreesC for 7 years 9 months. PMID- 19807045 TI - [Abundance of larvae and nymphs of the taiga tick Ixodes persuicatus (Acari: Ixodidae) on small mammals in the cut-over lands of the middle taiga subzone of Karelia]. AB - Data of long-term investigations (1998-2004) on the abundance of the taiga tick larvae and nymphs in the cut-over lands of different age in the middle taiga subzone of Karelia (62 degrees 04'S; 33 degrees 55'W) are presented. The investigation was carried out on three model cut-over lands of different age: 1) "young" cut-over land; age of cut-over in the beginning of investigation is 7 years; Betula-Deschampsia cespitosa-Agrostis tenuis; 2) "middle" cut-over land; age of cut-over is 12 years; Salix-Deschampsia cespitosa-Agrostis tenuis; 3) "old" cut-over land; age of cut-over 25 years; Alnus incana-Rubus idaeus-grass. The number of ticks was estimated by using common parasitological indices: prevalence, abundance, and index of feeding intensity (the tick abundance multiply by the number of small mammals per hundred traps-nights). In the beginning of investigation the "young" cut-over land was a typical meadow association. The lowest tick abundance was recorded here. That was a result of unfavorable abiotic conditions and low number of small mammals in the beginning of summer. "Middle" cut-over land is characterized by the highest number of the tick larvae, which is the evidence for high number of the hosts of tick imago. "Old" cut-over land has the optimum conditions for development of taiga ticks. High abundance of the ticks (larvae and nymphs) was recorded during the whole period of investigations. The number of preimaginal ticks is shown to be much higher in cut-over lands as compared with that in mixed and coniferous forests, due to the higher number of small mammals. PMID- 19807046 TI - [Hypertension 2009: what's new?]. PMID- 19807047 TI - [Is preeclampsia a new cardiovascular and end-stage renal diseases risk marker?]. AB - Is preeclampsia a new cardiovascular and end-stage renal diseases risk marker? Preeclampsia is a major obstetrical problem leading to high maternal and perinatal morbidity and mortality worldwide. It is characterized by hypertension and proteinuria occurring after 20 weeks of gestation. Increasing evidence indicates that preeclampsia is not just a pregnancy disease that resolves at the time of delivery, but represent a risk marker of cardiovascular and end-stage renal diseases later in life. The underlying link between preeclampsia and cardiovascular disease is unclear. Women with early onset/severe preeclampsia, recurrent preeclampsia, or preeclampsia with onset as a multipara appear to be at highest risk of cardiovascular disease later in life, including during the premenopausal period. PMID- 19807048 TI - [Hypertension in pregnancy]. AB - Hypertension in pregnancy Hypertension in pregnancy, whether chronic or recently diagnosed, is always a matter of concern for the general practitioner or the obstetrician. Even if this situation often evolves favorably, and although a "wait and see" attitude may be preferred to an aggressive one in such cases, one should also be aware of how dramatic the outcome may also be. As a matter of fact, what is considered as one of the most frequent complications of pregnancy can run out of control, a possibility which shouldn't be dismissed. In this article, we shall discuss the various strategies for managing this disorder. PMID- 19807049 TI - [Hypertension genetics: what have we learned so far?]. AB - Hypertension is a common, modifiable and heritable cardiovascular risk factor. Some rare monogenic forms of hypertension have been described, but the majority of patients suffer from "essential" hypertension, for whom the underlying pathophysiological mechanism is not clear. Essential hypertension is a complex trait, involving multiple genes and environmental factors. Recently, progress in the identification of common genetic variants associated with blood pressure and hypertension has been made thanks to large-scale international collaborative projects involving geneticists, epidemiologists, statisticians and clinicians. In this article, we review some basic genetic concepts and the main research methods used to study the genetics of hypertension, as well as selected recent findings in this field. PMID- 19807050 TI - [Hypertension in solid organ transplants]. AB - Hypertension is highly prevalent in transplantation and affects all type of organs. With the introduction of calcineurin inhibitors as immunosuppressive drugs, acute allograft rejection episodes have been significantly reduced and hence patient and allograft survival rates have dramatically improved. However, cardiovascular complications have become an important cause of morbidity and mortality. Treating cardiovascular risk factors such as diabetes, dyslipidemia and hypertension seems obvious, however in this population, there is little evidence for specific blood pressure targets, or for the best strategy to achieve blood pressure control. The aim of this article is to review the epidemiology and physiopathology of hypertension in transplant recipients as well as its clinical management. PMID- 19807051 TI - [Arterial hypertension: macrocirculation and microcirculation]. AB - The aim of the present report is to outline, in concise from, the changes in vascular structure which accompany hypertension. Consideration will be given to their potential contribution to hypertensive end organ damage. In so doing, it is important to consider both the macrovascular and microvascular levels, because interactions between them are presently believed to be critically important. The links between hypertension and the pathogenesis of arteriosclerosis fall outside the scope of this short review. PMID- 19807052 TI - [Depression in the elderly]. AB - Often under diagnosed and inappropriately treated, depression in the elderly is a major public health problem. Semiology is often atypical with especially frequent physical and hypochondriac complaints. Concomitant somatic affections, social and surrounding particularities of elderly persons make the diagnosis of depression particularity difficult to make. lts treatment calls for a careful pharmacological care and adaptation of the psychotherapeutic manageme PMID- 19807053 TI - [Everything is similar, certainly. Is everything equal?]. PMID- 19807054 TI - [Mysteries not resolved by A(H1N1)]. PMID- 19807055 TI - [Mr and/or Mrs Caster Semenya (2)]. PMID- 19807056 TI - [The "death tribunal" of Sarah Palin]. PMID- 19807057 TI - [Dialogues about the revolution]. PMID- 19807058 TI - Reversible dissolution of glutathione-mediated HgSe(x)S(1-x) nanoparticles and possible significance in Hg-Se antagonism. AB - A new pathway is proposed for the in vivo biomineralization of HgSe(x)S(1-x)(s) (0 < x < or = 1), which is thought to be the ultimate metabolic product responsible for the Hg-Se antagonism in biological systems. The pathway involves the reaction of inorganic Hg(II) with selenite in the presence of glutathione (GSH). The resulting GSH-mediated HgSe(x)S(1-x) nanoclusters are reversibly soluble depending on the pH, which might have played a role in the distribution pattern of HgSe(x)S(1-x)(s) in different tissues. The HgSe species involved and the nature of bonding were studied by UV-visible spectroscopy, infrared spectroscopy, microprobe scanning electron microscopy, transmission electron microscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. PMID- 19807059 TI - Synthesis and characterization of hydroxyl-functionalized caprolactone copolymers and their effect on adhesion, proliferation, and differentiation of human mesenchymal stem cells. AB - The aim of this study was to develop new hydrophilic polyesters for tissue engineering applications. In our approach, poly(benzyloxymethyl glycolide-co epsilon-caprolactone)s (pBHMG-CLs) were synthesized through melt copolymerization of epsilon-caprolactone (CL) and benzyl-protected hydroxymethyl glycolide (BHMG). Deprotection of the polymers yielded copolymers with pendant hydroxyl groups, poly(hydroxymethylglycolide-co-epsilon-caprolactone) (pHMG-CL). The synthesized polymers were characterized by GPC, NMR, and DSC techniques. The resulting copolymers consisting of up to 10% of HMG monomer were semicrystalline with a melting temperature above body temperature. Water contact angle measurements of polymeric films showed that increasing HMG content resulted in higher surface hydrophilicity, as evidenced from a decrease in receding contact angle from 68 degrees for PCL to 40 degrees for 10% HMG-CL. Human mesenchymal stem cells showed good adherence onto pHMG-CL films as compared to the more hydrophobic PCL surfaces. The cells survived and were able to differentiate toward osteogenic lineage on pHMG-CL surfaces. This study shows that the aforementioned hydrophilic polymers are attractive candidates for the design of scaffolds for tissue engineering applications. PMID- 19807060 TI - Structure of the amyloid-beta (1-42) monomer absorbed to model phospholipid bilayers: a molecular dynamics study. AB - The amyloid-beta (Abeta) peptide, the 39 to 43 amino acid peptide that plays a substantial role in Alzheimer's disease, has been shown to interact strongly with lipids both in vitro and in vivo. Abeta-lipid interactions have been proposed as a considerable factor in accelerating Abeta aggregation through the templating role of membranes in aggregation disorders. Previous work has shown that anionic lipids are able to significantly increase Abeta aggregation rate and induce a structural conversion in Abeta from a random coil to a beta-structure that is similar to the monomer structure observed in mature fibrils. However, it is unclear if this structural change occurs with the Abeta monomer because of direct interactions with the lipids or if the structural change results from protein protein interactions during oligomerization. We use extensive replica exchange molecular dynamics simulations of an Abeta monomer bound to a homogeneous model zwitterionic or anionic lipid bilayer. From these simulations, we do not observe any significant beta-structure formation except for a small, unstable beta hairpin formed on the anionic dioleylphosphatidylserine bilayer. Further, we see that the Asp23-Lys28 salt bridge that plays a role in beta-hairpin formation is not substantially formed on the bilayer surface and that Lys28 preferentially interacts with lipids when bound to the bilayer. These results suggest that the structural conversion seen in experiments are not due to the ordering of monomeric Abeta on the bilayer surface but are a result of protein-protein interactions enhanced by Abeta binding to the cell membrane. PMID- 19807061 TI - Accurate calculations of intermolecular interaction energies using explicitly correlated coupled cluster wave functions and a dispersion-weighted MP2 method. AB - Explicitly correlated coupled-cluster calculations of intermolecular interaction energies for the S22 benchmark set of Jurecka, Sponer, Cerny, and Hobza (Chem. Phys. Phys. Chem. 2006, 8, 1985) are presented. Results obtained with the recently proposed CCSD(T)-F12a method and augmented double-zeta basis sets are found to be in very close agreement with basis set extrapolated conventional CCSD(T) results. Furthermore, we propose a dispersion-weighted MP2 (DW-MP2) approximation that combines the good accuracy of MP2 for complexes with predominately electrostatic bonding and SCS-MP2 for dispersion-dominated ones. The MP2-F12 and SCS-MP2-F12 correlation energies are weighted by a switching function that depends on the relative HF and correlation contributions to the interaction energy. For the S22 set, this yields a mean absolute deviation of 0.2 kcal/mol from the CCSD(T)-F12a results. The method, which allows obtaining accurate results at low cost, is also tested for a number of dimers that are not in the training set. PMID- 19807062 TI - The ATP-dependent amide ligases DdaG and DdaF assemble the fumaramoyl-dipeptide scaffold of the dapdiamide antibiotics. AB - The enzymes DdaG and DdaF, encoded in the Pantoea agglomerans dapdiamide antibiotic biosynthetic gene cluster, when expressed in Escherichia coli, form the tandem amide bonds of the dapdiamide scaffold at the expense of ATP cleavage. DdaG uses fumarate, 2,3-diaminopropionate (DAP), and ATP to make fumaroyl-AMP transiently on the way to the N(beta)-fumaroyl-DAP regioisomer. Then DdaF acts as a second ATP-dependent amide ligase, but this enzyme cleaves ATP to ADP and P(i) during amide bond formation. However, DdaF will not accept N(beta)-fumaroyl-DAP; the enzyme requires the fumaroyl moiety to be first converted to the fumaramoyl half-amide in N(beta)-fumaramoyl-DAP. DdaF adds Val, Ile, or Leu to the carboxylate of fumaramoyl-DAP to make dapdiamide A, B, or C, respectively. Thus, to build the dapdiamide antibiotic scaffold, amidation must occur on the fumaroyl DAP scaffold, after DdaG action but before DdaF catalysis. This is an unusual instance of two ligases acting sequentially in untemplated amide bond formations using attack of substrate carboxylates at P(alpha) (AMP-forming) and then at P(gamma) (ADP-forming) of ATP cosubstrates. PMID- 19807063 TI - Microwave-assisted fluorous synthesis of a 1,4-benzodiazepine-2,5-dione library. AB - Fluorous displaceable linker-facilitated synthesis of 1,4-benzodiazepine-2,5 dione library has been developed. Perfluorooctanesulfonyl protected 4-hydroxy benzaldehydes were used as the limiting agent for Ugi four-component reactions to form condensed products. Postcondensation reactions of the Ugi products generated 1,4-benzodiazepine-2,5-dione ring skeleton. Microwave-assisted Suzuki coupling reactions removed the fluorous tag and introduced biaryl functionality to the benzodiazepine ring. The library scaffold has four points of substitution diversities. The fluorous tag facilitated the intermediate purifications using fluorous solid-phase extraction (F-SPE) and had no negative impact on the reactivity of the Ugi reactions and postcondensation reactions. PMID- 19807064 TI - Combinatorial materials research applied to the development of new surface coatings XIII: an investigation of polysiloxane antimicrobial coatings containing tethered quaternary ammonium salt groups. AB - High-throughput biological assays were used to develop structure - antimicrobial relationships for polysiloxane coatings containing chemically bound (tethered) quaternary ammonium salt (QAS) moieties. The QAS-functional polysiloxanes were derived from solution blends of a silanol-terminated polydimethylsiloxane, a trimethoxysilane-functional QAS (QAS-TMS), and methylacetoxysilane. Since the QAS moieties provide antimicrobial activity through interaction with the microorganism cell wall, most of the compositional variables that were investigated were associated with the chemical structure of the QAS-TMS. Twenty different QAS-TMS were synthesized for the study and the antimicrobial activity of sixty unique polysiloxane coatings derived from these QAS-TMS determined toward Escherichia coli , Staphylococcus aureus , and Candida albicans . The results of the study showed that essentially all of the compositional variables significantly influenced antimicrobial activity. Surface characterization of these moisture-cured coatings using atomic force microscopy as well as water contact angle and water contact angle hysteresis measurements indicated that the compositional variables significantly affected coating surface morphology and surface chemistry. Overall, compositional variables that produced heterogeneous surface morphologies provided the highest antimicrobial activity suggesting that the antimicrobial activity was primarily derived from the relationship between coating chemical composition and self-assembly of QAS moieties at the coating/air interface. Using data modeling software, a narrow region of the compositional space was identified that provided broad-spectrum antimicrobial activity. PMID- 19807065 TI - An optical conveyor for molecules. AB - Trapping single ions under vacuum allows for precise spectroscopy in atomic physics. The confinement of biological molecules in bulk water is hindered by the lack of comparably strong forces. Molecules have been immobilized to surfaces, however often with detrimental effects on their function. Here, we optically trap molecules by creating the microscale analogue of a conveyor belt: a bidirectional flow is combined with a perpendicular thermophoretic molecule drift. Arranged in a toroidal geometry, the conveyor accumulates a hundredfold excess of 5-base DNA within seconds. The concentrations of the trapped DNA scale exponentially with length, reaching trapping potential depths of 14 kT for 50 bases. The mechanism does not require microfluidics, electrodes, or surface modifications. As a result, the trap can be dynamically relocated. The optical conveyor can be used to enhance diffusion-limited surface reactions, redirect cellular signaling, observe individual biomolecules over a prolonged time, or approach single molecule chemistry in bulk water. PMID- 19807066 TI - Localization of Na+-K+ ATPases in quasi-native cell membranes. AB - Na(+)-K(+) ATPases have been observed and located by in situ AFM and single molecule recognition technique, topography and recognition imaging (TREC) that is a unique technique to specifically identify single protein in complex during AFM imaging. Na(+)-K(+) ATPases were well distributed in the inner leaflet of cell membranes with about 10% aggregations in total recognized proteins. The height of Na(+)-K(+) ATPases measured by AFM is in the range of 12-14 nm, which is very consistent with the cryoelectron microscopy result. The unbinding force between Na(+)-K(+) ATPases in the membrane and anti-ATPases on the AFM tip is about 80 pN with the apparent loading rate at 40 nN/s. Our results show the first visualization of an essential membrane protein, Na(+)-K(+) ATPase, in quasi native cell membranes and may be significant to reveal the interactions between Na(+)-K(+) ATPases and other membrane proteins at the molecular level. PMID- 19807067 TI - Nanoshells for surface-enhanced Raman spectroscopy in eukaryotic cells: cellular response and sensor development. AB - The application of gold nanoshells (NS) as a surface-enhanced Raman (SER) platform for intracellular sensing in NIH-3T3 fibroblast cells was studied by using a near-infrared Raman system. To show the feasibility of using these 151 +/ 5 nm sized solution-stable nanoparticles inside living cells, we investigated the uptake, cellular response, and the health of the cell population. We show that NS are taken up voluntarily and can be found in the cytosol by transmission electron microscopy (TEM), which also provides detailed information about location and immediate surrounding of the NS. The internalization into cells has been found to be independent of active cellular mechanisms, such as endocytosis, and can be suggested to be of passive nature. Uptake of NS into cells can be controlled, and cells show no increase in necrosis or apoptosis as a result; we show that NS-based intracytosolic SER spectra can be measured on biological samples using short acquisition times and low laser powers. We demonstrate its application using 4-mercaptobenzoic acid (4-MBA)-functionalized nanoshells as a pH sensor. PMID- 19807068 TI - Temperature-controlled syntheses of substituted 1,2,4-triazolelead(II) complexes: active lone pair and N-H...X (X = Cl, Br, I) hydrogen bonds. AB - Three isomorphous lead(II) complexes with a new {N(3)SX}-coordinated (X = Cl, Br, I) mode have been synthesized and characterized by X-ray diffraction. The structural analyses reveal that the coordination environment around the center metal is distinctly influenced by the stereochemical 6s(2) lone pair and N-H...X hydrogen bonds. Additionally, doubly deprotonated trzS(2-) (HtrzSH = 1,2,4 triazole-3-thiol) generated in situ via deamination and dehydrazination of ahtrzS(-) (ahtrzSH = 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole) was first observed in a semiconducting three-dimensional coordination network at higher reaction temperature. PMID- 19807069 TI - Platinum nanoparticle decorated silicon nanowires for efficient solar energy conversion. AB - High-density aligned n-type silicon nanowire (SiNW) arrays decorated with discrete 5-10 nm platinum nanoparticles (PtNPs) have been fabricated by aqueous electroless Si etching followed by an electroless platinum deposition process. Coating of PtNPs on SiNW sidewalls yielded a substantial enhancement in photoconversion efficiency and an apparent energy conversion efficiency of up to 8.14% for the PtNP-decorated SiNW-based photoelectrochemical solar cell using a liquid electrolyte containing Br(-)/Br(2) redox couple. The results demonstrate PtNP-decorated SiNWs to be a promising hybrid system for solar energy conversion. PMID- 19807070 TI - Design, synthesis, and testing of difluoroboron-derivatized curcumins as near infrared probes for in vivo detection of amyloid-beta deposits. AB - Amyloid-beta (Abeta) deposits have been identified as key players in the progression of Alzheimer's disease (AD). Recent evidence indicates that the deposits probably precede and induce the neuronal atrophy. Therefore, methods that enable monitoring the pathology before clinical symptoms are observed would be beneficial for early AD detection. Here, we report the design, synthesis, and testing of a curcumin-derivatized near-infrared (NIR) probe, CRANAD-2. Upon interacting with Abeta aggregates, CRANAD-2 undergoes a range of changes, which include a 70-fold fluorescence intensity increase, a 90 nm blue shift (from 805 to 715 nm), and a large increase in quantum yield. Moreover, this probe also shows a high affinity for Abeta aggregates (K(d) = 38.0 nM), a reasonable log P value (log P = 3), considerable stability in serum, and a weak interaction with albumin. After intravenous injection of this probe, 19-month-old Tg2576 mice exhibited significantly higher relative signal than that of the control mice over the same period of time. In summary, CRANAD-2 meets all the requirements for a NIR contrast agent for the detection of Abeta plaques both in vitro and in vivo. Our data point toward the feasibility of monitoring the progress of the disease by NIR imaging with CRANAD-2. In addition, we believe that our probe could be potentially used as a tool for drug screening. PMID- 19807071 TI - Surface and bulk aspects of mixed oxide catalytic nanoparticles: oxidation and dehydration of CH(3)OH by polyoxometallates. AB - The molecular structures and surface chemistry of mixed metal oxide heteropolyoxo vanadium tungstate (H(3+x)PW(12-x)V(x)O(40) with x = 0, 1, 2, and 3) Keggin nanoparticles (NPs), where vanadium is incorporated into the primary Keggin structure, and supported VO(x) on tungstophosphoric acid (TPA, H(3)PW(12)O(40)), where vanadium is present on the surface of the Keggin unit, were investigated with solid-state magic angle spinning (51)V NMR, FT-IR, in situ Raman, in situ UV vis, CH(3)OH temperature-programmed surface reaction (TPSR), and steady-state methanol oxidation. The incorporated VO(x) unit possesses one terminal V horizontal lineO bond, four bridging V-O-W/V bonds, and one long V-O-P bond in the primary Keggin structure, and the supported VO(x) unit possesses a similar coordination in the secondary structure under ambient conditions. The specific redox reaction rate for VO(x) in the Keggin primary structure is comparable to that of bulk V(2)O(5) and the more active supported vanadium oxide catalysts. The specific acidic reaction rate for the WO(x) in the TPA Keggin, however, is orders of magnitude greater than found for bulk WO(3), supported tungsten oxide catalysts, and even the highly acidic WO(3)-ZrO(2) catalyst synthesized by coprecipitation of ammonium metatungstate and ZrO(OH)(2). From CH(3)OH-TPSR and in situ Raman spectroscopy it was found that incorporation of vanadium oxide into the primary Keggin structure is also accompanied by the formation of surface VO(x) species at secondary sites on the Keggin outer surface. Both CH(3)OH-TPSR and steady-state methanol oxidation studies demonstrated that the surface VO(x) species on the Keggin outer surface are significantly less active than the VO(x) species incorporated into the primary Keggin structure. The presence of the less active surface VO(x) sites in the Keggins, thus, decreases the specific reaction rates for both methanol oxidation and methanol dehydration. During methanol oxidation/dehydration (O(2)/CH(3)OH = 2.17, T = 225 degrees C), in situ UV-vis diffuse reflectance spectroscopy revealed that vanadium oxide is primarily present as the V(+5) cation, which reflects the Mars-van Krevelen redox mechanism and rapid reoxidation by molecular O(2). The bulk TPA Keggin structure becomes more disordered and less thermally stable as the vanadium content increases. Although surface polyaromatic carbon forms during methanol oxidation on the Keggin surfaces, its influence on the reaction kinetics seems minimal as the carbon content diminishes as the vanadium oxide content increases and the reaction temperature is raised. No relationships were found between the electronic structure (UV-vis E(g) values) and TOF(redox) and TOF(acid) (TOF = turnover frequency) kinetics, which reflect the complexity of H(3+x)PW(12 x)V(x)O(40) Keggins. The overall catalytic performance of the H(3+x)PW(12 x)V(x)O(40) Keggin materials results from a complex interplay among the presence of redox vanadium (as secondary surface VO(x) species and substituted VO(x) sites in the primary Keggin NP structure), structural disorder of the Keggin NPs, exposed surface acid and redox sites, and coke deposition. These new insights reveal that the Keggin heteropolyoxometallates are much more complex than originally thought and that care must be taken in using Keggins as model mixed metal oxide NPs in catalytic kinetic and theoretical studies because their surface and bulk structures are dynamic under the reaction conditions. PMID- 19807072 TI - Molecular characterization of flubendiamide sensitivity in the lepidopterous ryanodine receptor Ca(2+) release channel. AB - Flubendiamide is a benzenedicarboxamide derivative that shows selective insecticidal activity against lepidopterous insects. The specific modulatory effects of flubendiamide on ryanodine binding in insect muscle microsomal membranes suggest that the ryanodine receptor (RyR) Ca(2+) release channel is a primary target of flubendiamide. However, the molecular mechanisms underlying the species-specific action of flubendiamide are unclear. We have cloned cDNA encoding a novel RyR from the lepidopterous silkworm RyR (sRyR) and tested the sensitivity to flubendiamide of the recombinant sRyR in HEK293 cells. Confocal localization studies and Ca(2+) imaging techniques revealed that sRyRs form Ca(2+) release channels in the endoplasmic reticulum. Importantly, flubendiamide induced release of Ca(2+) through the sRyR, but not through the rabbit RyR isoforms. Photoaffinity labeling of sRyR deletion mutants using a photoreactive derivative revealed that flubendiamide is mainly incorporated into the transmembrane domain (amino acids 4111-5084) of the sRyR. The rabbit cardiac muscle isoform RyR2 (rRyR2) and the RyR mutant carrying a replacement of the transmembrane domain (residues 4084-5084) with its counterpart sequence from rRyR2 (residues 3936-4968) were not labeled by the photoreactive compound. This replacement in the sRyR significantly impaired the responses to flubendiamide but only marginally reduced the sensitivity to caffeine, a general RyR activator. Furthermore, deletion of the N-terminal sequence (residues 183-290) abolished the responses of the sRyR to flubendiamide but not the sensitivity to caffeine. Our results suggest that the transmembrane domain plays an important role in the formation of an action site for flubendiamide, while the N-terminus is a structural requirement for flubendiamide-induced activation of the sRyR. PMID- 19807073 TI - Rh(II)-Catalyzed skeletal reorganization of 1,6- and 1,7-enynes through electrophilic activation of alkynes. AB - The skeletal reorganization of 1,6- and 1,7-enynes leading to 1-vinylcycloalkenes using Rh(II) as a catalyst is reported. Two possible isomers of 1 vinylcycloalkenes, type I and type II, can be obtained, the ratio of which are highly dependent on the substitution pattern of the enynes used. Formation of type I compounds involves a single cleavage of a C-C double bond, the product of which is identical to that of enyne metathesis. In contrast, the formation of type II compounds involves the double cleavage of both the C-C double and triple bonds, which has an anomalous bond connection. The presence of both a phenyl group at the alkyne carbon and a methyl group at the internal alkene carbon facilitates the formation of type II compounds. The electronic and steric nature of the substituents on the aromatic ring also affects the ratio of type I and type II. The nature of a tether also has a significant effect on the course of the reaction. Experimental evidence for the intermediacy of a cyclopropyl rhodium carbenoid, a key intermediate in the skeletal reorganization of enynes, is also reported. In addition to the skeletal reorganization of enynes, Rh(II) complexes were found to have a high catalytic activity for some cycloisomerization reactions of alkyne derivatives, including the bicyclization of enynes to bicyclo[4.1.0]heptene derivatives and the cyclization of alkynylfurans to phenol derivatives. PMID- 19807075 TI - Lanthanide-containing polymer microspheres by multiple-stage dispersion polymerization for highly multiplexed bioassays. AB - We describe the synthesis and characterization of metal-encoded polystyrene microspheres by multiple-stage dispersion polymerization with diameters on the order of 2 mum and a very narrow size distribution. Different lanthanides were loaded into these microspheres through the addition of a mixture of lanthanide salts (LnCl(3)) and excess acrylic acid (AA) or acetoacetylethyl methacrylate (AAEM) dissolved in ethanol to the reaction after about 10% conversion of styrene, that is, well after the particle nucleation stage was complete. Individual microspheres contain ca. 10(6)-10(8) chelated lanthanide ions, of either a single element or a mixture of elements. These microspheres were characterized one-by-one utilizing a novel mass cytometer with an inductively coupled plasma (ICP) ionization source and time-of-flight (TOF) mass spectrometry detection. Microspheres containing a range of different metals at different levels of concentration were synthesized to meet the requirements of binary encoding and enumeration encoding protocols. With four different metals at five levels of concentration, we could achieve a variability of 624, and the strategy we report should allow one to obtain much larger variability. To demonstrate the usefulness of element-encoded beads for highly multiplexed immunoassays, we carried out a proof-of-principle model bioassay involving conjugation of mouse IgG to the surface of La and Tm containing particles and its detection by an antimouse IgG bearing a metal-chelating polymer with Pr. PMID- 19807076 TI - New syntheses of E7389 C14-C35 and halichondrin C14-C38 building blocks: double inversion approach. AB - With sequential use of catalytic asymmetric Cr-mediated coupling reactions, E7389 C14-C35 and halichondrin C14-C38 building blocks have been stereoselectively synthesized. The C19-C20 bond is first formed via the catalytic asymmetric Ni/Cr mediated coupling, i.e., 8 + 9 --> 10 (90%; dr = 22:1), in which vinyl iodide 8 is used as the limiting substrate. The C23-C24 bond is then formed via the catalytic asymmetric Co/Cr-mediated coupling, i.e., 13 + 14 --> 4 (82%; dr = 22:1), in which the alkyl-iodide bond in 14 is selectively activated over the vinyl-iodide bond. The catalytic asymmetric Ni/Cr-mediated reaction is employed to couple C14-C26 segment 19 with E7389 C27-C35 segment 20 (91%; dr = >55:1). In this synthesis, the C23-O bond is stereoselectively constructed via a double inversion process, i.e., 21 --> 22, to furnish E7389 C14-C35 building block 22 in 84% yield. The same synthetic sequence has been employed to synthesize halichondrin C14-C38 building block 18b, i.e., 16a + 19 --> 18b. PMID- 19807077 TI - New syntheses of E7389 C14-C35 and halichondrin C14-C38 building blocks: reductive cyclization and oxy-Michael cyclization approaches. AB - Cr-mediated coupling reactions are usually achieved with a slight excess of a given nucleophile. To develop a cost-effective use of this process, two different approaches have been studied. The first approach depends on two consecutive catalytic asymmetric Cr-mediated couplings, with use of coupling partners purposely being of unbalanced molecular size and complexity. The second approach rests on the success in identifying the nucleophile, which allows us to achieve the coupling satisfactorily with a 1:1 molar ratio of the coupling partners. The C23-O bond is stereospecifically constructed via reductive cyclization of the oxonium ion, or oxy-Michael cyclization. Both syntheses have a high overall efficiency: E7389 C14-C35 and halichondrin C14-C38 building blocks have been synthesized from the corresponding C27-C35 and C27-C38 aldehydes, respectively, in high overall yields with an excellent stereoselectivity. Because of operational simplicity, the synthesis outlined herein appears to be well suited for scaling. PMID- 19807079 TI - Cage hydrocarbons derived from dibenzosuberenone. AB - The reinvestigation of the synthesis of the fascinating cage compound 2 reveals an incorrect structure of photodimer 2, as well as of the putatively isolated intermediate 9. These products have to be reassigned as monomeric pyramidalized alkene 3 and spirocyclic dichloride 10, respectively. PMID- 19807078 TI - An electrochemical sensor for single nucleotide polymorphism detection in serum based on a triple-stem DNA probe. AB - We report here an electrochemical approach that offers, for the first time, single-step, room-temperature single nucleotide polymorphism (SNP) detection directly in complex samples (such as blood serum) without the need for target modification, postwashing, or the addition of exogenous reagents. This sensor, which is sensitive, stable, and reusable, is comprised of a single, self complementary, methylene blue-labeled DNA probe possessing a triple-stem structure. This probe takes advantage of the large thermodynamic changes in enthalpy and entropy that result from major conformational rearrangements that occur upon binding a perfectly matched target, resulting in a large-scale change in the faradaic current. As a result, the discrimination capabilities of this sensor greatly exceed those of earlier single- and double-stem electrochemical sensors and support rapid (minutes), single-step, reagentless, room-temperature detection of single nucleotide substitutions. To elucidate the theoretical basis of the sensor's selectivity, we present a comparative thermodynamic analysis among single-, double-, and triple-stem probes. PMID- 19807080 TI - Cs(2)K(UO)(2)Si(4)O(12): a mixed-valence uranium(IV,V) silicate. AB - The first mixed-valence uranium(IV,V) silicate is synthesized under high temperature, high-pressure hydrothermal conditions. The structure contains chains of corner-sharing U(IV,V)O(6) octahedra which are interconnected by Si(4)O(12) four-membered rings to form a 3-D framework. XPS and XANES spectra were measured to identify the valence state of uranium. PMID- 19807081 TI - Fine tuning reactivity: synthesis and isolation of 1,2,3,12b tetrahydroimidazo[1,2-f]phenanthridines. AB - A facile route for the synthesis and isolation of 1,2,3,12b-tetrahydroimidazo[1,2 f]phenanthridines (TIPs) has been developed. The heterocycle is a reactive intermediate in the three-step cascade synthesis of 2,3-dihydro-1H-imidazo[1,2 f]phenanthridinium cations (DIPs), a biologically active DNA intercalating framework; however, the intermediate has previously only been characterized in situ. Derivatization of the structure at the imidazo-N position controls the reactivity of the intermediate with respect to electronic potential and pK(a) allowing isolation of a selection of TIP structures. Correlations between these parameters and reaction outcome have been made, and other influences such as steric and solvent effects have also been investigated. PMID- 19807082 TI - Divalent metal ion triggered activity of a synthetic antimicrobial in cardiolipin membranes. AB - One member of a prototypical class of antimicrobial oligomers was used to study pore formation in cardiolipin-rich membranes. Both vesicle dye-leakage assays and small-angle X-ray scattering were used to study bilayer remodeling. The results indicate that the presence of negative intrinsic curvature lipids is essential for pore formation by this class of molecules: In Gram-positive bacteria, cardiolipin and divalent metal cations like Ca(2+) and Mg(2+) are needed. This is consistent with the role of phosphatidylethanolamine (PE) lipid in Gram-negative bacteria, where antimicrobial activity is dependent on the negative intrinsic curvature of PE rather than a specific interaction with PE. PMID- 19807083 TI - Active metal template synthesis of [2]catenanes. AB - The synthesis of [2]catenanes by single macrocyclization and double macrocyclization strategies using Cu(I) ions to catalyze covalent bond formation while simultaneously acting as the template for the mechanically interlocked structure is reported. These "active metal template" strategies employ appropriately functionalized pyridine ether or bipyridine ligands and either the CuAAC "click" reaction of azides with terminal alkynes or the Cu(I)-mediated Cadiot-Chodkiewicz heterocoupling of an alkyne halide with a terminal alkyne. Using one macrocyclic and one acyclic building block, heterocircuit (the rings are constitutionally different) [2]catenanes are produced via the single macrocyclization route in up to 53% yield by optimizing the reaction conditions and relative stoichiometry of the starting materials. Alternatively, with the active template CuAAC reaction, a single acyclic unit can be used to form a homocircuit (two identical rings) [2]catenane in 46% yield through a one-pot, double macrocyclization, procedure. Remarkably, <7% of the corresponding noninterlocked macrocycle is isolated from this reaction, indicating the efficacy of Cu(I) as both a template for the catenane and a catalyst for covalent bond formation in the reaction. PMID- 19807084 TI - Memory effects of diblock copolymer brushes and mixed brushes. AB - Memory effects of microphase segregation in diblock copolymer brushes and binary mixed homopolymer brushes exposed to solvents of different quality and selectivity are studied using Single-Chain-in-Mean-Field (SCMF) simulations. We gauge these memory effects by a fluctuation memory measure, reflecting the correlation between the quenched fluctuations of grafting points and the microphase-separated morphology, and a domain memory measure, quantifying the correlation between surface morphologies during cyclic exposure to different solvents. The fluctuation and domain memory measures are closely correlated, and both of them have their root in the broken translational symmetry of the distribution of grafting points. They become stronger upon increasing the fluctuations of the grafting points. The effects of solvent quality and selectivity, grafting density, and composition of brushes on the memory measures are discussed. PMID- 19807085 TI - Left- and right-handed alpha-helical turns in homo- and hetero-chiral helical scaffolds. AB - Proteins typically consist of right-handed alpha helices, whereas left-handed alpha helices are rare in nature. Peptides of 20 amino acids or less corresponding to protein helices do not form thermodynamically stable alpha helices in water away from protein environments. The smallest known water-stable right- (alpha(R)) and left- (alpha(L)) handed alpha helices are reported, each stabilized in cyclic pentapeptide units containing all L- or all D-amino acids. Homochiral decapeptides comprising two identical cyclic pentapeptides (alpha(R)alpha(R) or alpha(L)alpha(L)) are continuous alpha-helical structures that are extremely stable to denaturants, degradative proteases, serum, and additives like TFE, acid, and base. Heterochiral decapeptides comprising two different cyclic pentapeptides (alpha(L)alpha(R) or alpha(R)alpha(L)) maintain the respective helical handedness of each monocyclic helical turn component but adopt extended or bent helical structures depending on the solvent environment. Adding TFE to their aqueous solutions caused a change to bent helical structures with slightly distorted N-terminal alpha(R) or alpha(L)-helical turns terminated by a Schellman-like motif adjacent to the C-terminal alpha(L) or alpha(R)-turn. This hinge-like switching between structures in response to an external cue suggests possible uses in larger structures to generate smart materials. The library of left- and right-handed 1-3 turn alpha-helical compounds reported herein project their amino acid side chains into very different regions of 3D space, constituting a unique and potentially valuable class of novel scaffolds. PMID- 19807086 TI - Laboratory evolution of robust and enantioselective Baeyer-Villiger monooxygenases for asymmetric catalysis. AB - The Baeyer-Villiger Monooxygenase, Phenylacetone Monooxygenase (PAMO), recently discovered by Fraaije, Janssen, and co-workers, is unusually thermostable, which makes it a promising candidate for catalyzing enantioselective Baeyer-Villiger reactions in organic chemistry. Unfortunately, however, its substrate scope is very limited, reasonable reaction rates being observed essentially only with phenylacetone and similar linear phenyl-substituted analogs. Previous protein engineering attempts to broaden the range of substrate acceptance and to control enantioselectivity have been met with limited success, including rational design and directed evolution based on saturation mutagenesis with formation of focused mutant libraries, which may have to do with complex domain movements. In the present study, a new approach to laboratory evolution is described which has led to mutants showing unusually high activity and enantioselectivity in the oxidative kinetic resolution of a variety of 2-aryl and 2-alkylcyclohexanones which are not accepted by the wild-type (WT) PAMO and of a structurally very different bicyclic ketone. The new strategy exploits bioinformatics data derived from sequence alignment of eight different Baeyer-Villiger Monooxygenases, which in conjunction with the known X-ray structure of PAMO and induced fit docking suggests potential randomization sites, different from all previous approaches to focused library generation. Sites harboring highly conserved proline in a loop of the WT are targeted. The most active and enantioselective mutants retain the high thermostability of the parent WT PAMO. The success of the "proline" hypothesis in the present system calls for further testing in future laboratory evolution studies. PMID- 19807087 TI - Ferromagnetic CoPt3 nanowires: structural evolution from fcc to ordered L1(2). AB - This investigation demonstrates the magnetic properties and nanostructures of CoPt(3) wire arrays that were fabricated by electrodeposition using a porous alumina template. The X-ray absorption analysis clearly verified the occurrence of a phase transition in CoPt(3) nanowires. This phase transition significantly influences the magnetic properties and enhances coercivity and squareness. The phase transition of CoPt(3) nanowires was from a random alloy distribution to anisotropically ordered CoPt(3) (L1(2)). The thermally induced phase transition of CoPt(3) nanowires to ordered L1(2) CoPt(3) through a "cluster-in-cluster" intermediate state via interdiffusion processes is revealed. The mechanism exhibited by these CoPt(3) nanowires is proposed to explain the strong correlation between their magnetic character and their atomic distribution. PMID- 19807089 TI - Impacts of the location and number of [Cu(bpy)(2)](2+) cross-links on the emission photodynamics of [Ru(bpy)(3)](2+) with pendant oligo(aminoethylglycine) chains. AB - Multifunctional aminoethylglycine (aeg) derivatized [Ru(bpy)(3)](2+) complexes with pendant bipyridine (bpy) ligands coordinate Cu(2+) to form coordinative chain cross-links in a "hairpin loop" motif. In this paper, we report the synthesis and characterization of a series of Ru aeg hairpins in which the relative aeg chain length and number of pendant bpy ligands is varied. Reaction of each of these with Cu(2+) is monitored using spectrophotometric emission titrations to determine the binding stoichiometry. Coordination of Cu(2+) causes quenching of the emissive excited state Ru species; the degree of quenching efficiency depends on the location and number of coordinated Cu ions. The heterometallic structures are fully characterized, and using the quantum yields and time-resolved emission following excitation of the Ru complexes in deoxygenated solutions, the radiative (k(r)) and nonradiative (k(nr)) relaxation rates are compared. These data reveal only a shallow decrease in k(nr) with increasing distance between the Ru and Cu complexes. Activation energies, determined from temperature dependent studies of the time-resolved emission, also increase as the Ru-Cu separation increases, resulting in the smaller nonradiative rates. Together, these data are suggestive of excited state electron transfer as the quenching mechanism and demonstrate that metal coordination self-assembles structures made from modular artificial amino acids can provide controlled arrangements of chromophores, electron donors, and electron acceptors to shuttle electrons in a new approach for mimicking photosynthesis. PMID- 19807088 TI - Multilayer DNA origami packed on a square lattice. AB - Molecular self-assembly using DNA as a structural building block has proven to be an efficient route to the construction of nanoscale objects and arrays of increasing complexity. Using the remarkable "scaffolded DNA origami" strategy, Rothemund demonstrated that a long single-stranded DNA from a viral genome (M13) can be folded into a variety of custom two-dimensional (2D) shapes using hundreds of short synthetic DNA molecules as staple strands. More recently, we generalized a strategy to build custom-shaped, three-dimensional (3D) objects formed as pleated layers of helices constrained to a honeycomb lattice, with precisely controlled dimensions ranging from 10 to 100 nm. Here we describe a more compact design for 3D origami, with layers of helices packed on a square lattice, that can be folded successfully into structures of designed dimensions in a one-step annealing process, despite the increased density of DNA helices. A square lattice provides a more natural framework for designing rectangular structures, the option for a more densely packed architecture, and the ability to create surfaces that are more flat than is possible with the honeycomb lattice. Thus enabling the design and construction of custom 3D shapes from helices packed on a square lattice provides a general foundational advance for increasing the versatility and scope of DNA nanotechnology. PMID- 19807090 TI - Conformational analysis of macrocycles: finding what common search methods miss. AB - As computational drug design becomes increasingly reliant on virtual screening and on high-throughput 3D modeling, the need for fast, robust, and reliable methods for sampling molecular conformations has become greater than ever. Furthermore, chemical novelty is at a premium, forcing medicinal chemists to explore more complex structural motifs and unusual topologies. This necessitates the use of conformational sampling techniques that work well in all cases. Here, we compare the performance of several popular conformational search algorithms on three broad classes of macrocyclic molecules. These methods include Catalyst, CAESAR, MacroModel, MOE, Omega, Rubicon and two newer self-organizing algorithms known as stochastic proximity embedding (SPE) and self-organizing superimposition (SOS) that have been developed at Johnson & Johnson. Our results show a compelling advantage for the three distance geometry methods (SOS, SPE, and Rubicon) followed to a lesser extent by MacroModel. The remaining techniques, particularly those based on systematic search, often failed to identify any of the lowest energy conformations and are unsuitable for this class of structures. Taken together with our previous study on drug-like molecules (Agrafiotis, D. K.; Gibbs, A.; Zhu, F.; Izrailev, S.; Martin, E. Conformational Sampling of Bioactive Molecules: A Comparative Study. J. Chem. Inf. Model., 2007, 47, 1067-1086), these results suggest that SPE and SOS are two of the most robust and universally applicable conformational search methods, with the latter being preferred because of its superior speed. PMID- 19807091 TI - Assessment of the multiphase interaction between a membrane disrupting peptide and a lipid membrane. AB - Although modeling and experimental approaches to probe antimicrobial peptides lipid membranes interaction have already been reported, quantitative evaluation of the whole process, including full dissolution of the lipid, is still missing. We report on the real-time assessment of the entire set of stages of melittin membrane interaction, based on surface plasmon resonance (SPR) measurements, using supported lipid matrices on L1 sensors and long peptide injections. We advance a mathematical model which comprises a set of coupled kinetic equations and relates via the transfer matrix the evolution of lipid and peptide concentrations with the SPR sensorgram. Upon fitting the sensorgrams of melittin injections on POPC lipid matrices, in agreement with literature data, the model provides: association and dissociation rates, concentration thresholds, and evolution within each interacting layer of lipid and peptide concentrations as well as of peptide to lipid ratios. The proposed model combined with appropriate experimental protocols adds new depths to SPR investigation of peptide-lipid interaction offering a quantitative platform for research and controlled design of improved antimicrobial peptides. A wider applicability for quantitative assessment of other pore forming compounds on different lipid matrices is suggested. PMID- 19807092 TI - Substituent effects on the thermodynamic stability of imines formed from glycine and aromatic aldehydes: implications for the catalytic activity of pyridoxal-5' phosphate. AB - Equilibrium constants for addition of glycine to substituted benzaldehydes to form the corresponding imines and pK(a)'s for ionization of the iminium ions were determined by (1)H NMR analysis in D(2)O. The introduction of a phenoxide anion substituent into the aromatic ring of benzaldehyde leads to a substantial increase in the pK(a) of the iminium ion from 6.3 to 10.2 for p hydroxybenzaldehyde and to 12.1 for salicylaldehyde. An analysis of the differential effect of ortho- versus para-substitution shows that the iminium ion to salicylaldehyde is stabilized by an intramolecular hydrogen bond in aqueous solution, with an estimated energy ca. 3 kcal/mol larger than can be accounted for by a simple electrostatic interaction. A comparison of the o-O(-) substituent effect on the acidity of the iminium ions of glycine to benzaldehyde and 4 pyridine-carboxaldehyde provides evidence for the existence of an internal hydrogen bond of similar strength in pyridoxal 5'-phosphate (PLP) iminium ions in water. The effects of other ring substituents on the stability of PLP iminium ions are discussed. PMID- 19807093 TI - Characterization of nonlinear background components in voltammetry by use of large amplitude periodic perturbations and fourier transform analysis. AB - Under most experimental conditions, a distinctly nonlinear background current is encountered in all forms of voltammetry which arises from the potential dependence of the capacitance. The nonlinear background current has been successfully modeled under large amplitude sinusoidal ac voltammetric conditions with a fourth order polynomial. The model was applied to a dummy cell containing a nonideal ceramic capacitor and commonly used electrodes. The nonlinearity in behavior of the background capacitance is particularly significant when considering the discrimination between the Faradaic and background contributions in the higher order harmonics resolved in ac voltammetry by Fourier transform inverse Fourier transform approaches and in the simulation of the background current and hence double-layer capacitance as a function of potential. Typically, measurable background current under large amplitude conditions is detectable in the dc and fundamental to fourth harmonic components in large amplitude ac voltammetry. For analytical purposes, this background current can be corrected on a per harmonic basis without the need for any model. Background correction has been successfully applied to the first four harmonics for the oxidation of ferrocenemonocarboxylic acid over the concentration range of 5-500 microM in aqueous 0.5 M NaCl solution. PMID- 19807094 TI - The effects of C-terminal modifications on the opioid activity of [N benzylTyr(1)]dynorphin A-(1-11) analogues. AB - Structural modifications affecting the efficacy of analogues of the endogenous opioid peptide dynorphin (Dyn) A have focused on the N-terminal "message" sequence based on the "message-address" concept. To test the hypothesis that changes in the C-terminal "address" domain could affect efficacy, modified amino acids and cyclic constraints were incorporated into this region of the partial agonist [N-benzylTyr(1)]Dyn A-(1-11). Modifications in the C-terminal domain of [N-benzylTyr(1)]Dyn A-(1-11)NH(2) resulted in increased kappa opioid receptor (KOR) affinity for all of the linear analogues but did not affect the efficacy of these peptides at KOR. Cyclization between positions 5 and 8 yielded [N benzylTyr(1),cyclo(d-Asp(5),Dap(8))]Dyn A-(1-11)NH(2) (zyklophin, 13) ( J. Med. Chem. 2005 , 48 , 4500 - 4503 ) with high selectivity for KOR. In contrast to the linear peptides, this peptide exhibits negligible efficacy in the adenylyl cyclase (AC) assay and is a KOR antagonist. These data are consistent with our hypothesis that appropriate modifications in the "address" domain of Dyn A analogues may affect efficacy. PMID- 19807095 TI - Sustained activation of glial cell epidermal growth factor receptor by bis(thiosemicarbazonato) metal complexes is associated with inhibition of protein tyrosine phosphatase activity. AB - Bis(thiosemicarbazonato) metal complexes (M(II)(btsc)) have demonstrated potential neuroprotective activity in cell and animal models of Alzheimer's disease (AD). Metal complexes can activate the epidermal growth factor receptor (EGFR), leading to inhibition of amyloid peptide accumulation in neuronal cells. As glial cells also have an important role in modulating neuronal health and survival in AD, we examined the effect of M(II)(btsc) on activity of EGFR in an astroglial cell line. Our findings reveal potent activation of glial EGFR by glyoxalbis(N(4)-methylthiosemicarbazonato)Cu(II)] (Cu(II)(gtsm)). Activation of EGFR by Cu(II)(gtsm) involved phosphorylation of multiple tyrosine residues and was mediated by a cognate ligand-independent process involving M(II)(btsc) inhibition of protein tyrosine phosphatase (PTP) activity. EGFR activation resulted in release of growth factors and cytokines with potential modulatory effects on neuronal function. These studies provide an important insight into the mechanism of action of a neuroprotective M(II)(btsc) and provide a basis for future studies into this novel approach to AD therapy. PMID- 19807096 TI - Effects of dietary selenomethionine supplementation on growth performance, meat quality and antioxidant property in yellow broilers. AB - This study was conducted to investigate the effects of dietary selenomethionine (Se-Met) supplementation on growth performance, meat quality and antioxidant property in male broilers. A total of 800 43-day-old Lingnan yellow male broilers were randomly allotted to 5 dietary treatments with four replicates of 40 birds for a period of 3 weeks ad libitum. Final BW and weight gain of birds significantly increased by Se-Met supplementation at the 0.225 mg/kg level (P < 0.05). The addition of Se-Met significantly decreased drip loss, lightness value, and elevated pH value of meat (p < 0.05). Adding sodium selenite (SS) only increased pH value of meat (p < 0.05). In plasma, supplemental Se-Met at 0.225 mg/kg level increased total antioxidant capability (T-AOC), glutathione peroxidase (GPX), total superoxide dismutase (T-SOD), catalase (CAT) activities, glutathione (GSH) concentration (p < 0.05), and decreased malondialdehyde production (p < 0.05), compared with the control and broilers fed SS diet. In breast muscle, the addition of Se-Met significantly elevated T-AOC, GPX, T-SOD, CAT activities, contents of metallothionein and GSH (p < 0.05), and reduced carbonyl protein content (p < 0.05). While compared with broilers fed SS diet, supplemental 0.225 mg/kg Se-Met increased T-AOC, GPX, CAT activities, and GSH content (p < 0.05). Therefore, dietary Se-Met supplementation could improve growth performance and meat quality by enhancing antioxidative capacity in broilers compared with SS. PMID- 19807097 TI - Survey of 3-alkyl-2-methoxypyrazine content of South African Sauvignon blanc wines using a novel LC-APCI-MS/MS method. AB - An LC-MS/MS method for the trace-level determination of 3-alkyl-2 methoxypyrazines in Sauvignon blanc wines is described. 3-Isobutyl-2 methoxypyrazine (IBMP), 3-isopropyl-2-methoxypyrazine (IPMP) and 3-sec-butyl-2 methoxypyrazine (SBMP) were analyzed by reversed phase liquid chromatography coupled to atmospheric pressure chemical ionization, as electrospray ionization was found to suffer from matrix quenching effects. A sample preparation method involving distillation of wine followed by solvent extraction and sufficient preconcentration was developed. Limits of detection and quantification for all three analytes were 0.03 ng/L and 0.10 ng/L, respectively, making the method more sensitive than gas chromatographic methods. IBMP was found to be the most abundant congener in South African Sauvignon blanc wines, with concentrations varying between 0.40 and 44 ng/L in 575 samples. IPMP and SBMP levels varied from <0.03 to 3.9 and <0.03 to 3.2 ng/L, respectively. Statistical investigation indicated no clear correlation between methoxypyrazine content and either geographical origin or vintage. The method was also successfully applied for the quantitation of IBMP in five additional South African wine varieties, including three red wine cultivars. The developed method represents a powerful new tool for the in-depth investigation of these important wine aroma constituents. PMID- 19807098 TI - Nondigestible saccharides suppress the bacterial degradation of quercetin aglycone in the large intestine and enhance the bioavailability of quercetin glucoside in rats. AB - Contribution of intestinal bacterial degradation of quercetin aglycone to the promotive effects of fructooligosaccharides and di-D-fructose anhydride III (DFAIII) on quercetin-3-O-beta-glucoside (Q3G) bioavailability was examined. Male Sprague-Dawley rats were fed 0.68% Q3G diets with or without 1.5% or 3% oligosaccharides for 2 weeks. Blood levels and urinary excretion of quercetin and methylquercetin conjugates, measured by methanol extraction and LC-MS analyses, were dose-dependently and adaptively increased by the oligosaccharide supplementation with increasing cecal fermentation (Experiment 1). Degradation of Q3G and quercetin aglycone by cecal bacteria in oligosaccharide-fed rats was much lower than that in the control rats using an anaerobic culture system (Experiment 2). Using the ligated intestinal sacs of anesthetized rats, we found that the cecum possessed high absorptive capacity for quercetin derivatives (Experiment 3). These results demonstrate that feeding of the oligosaccharides strongly suppresses the bacterial degradation of quercetin aglycone in the cecum, thus largely contributing to the increased bioavailability of Q3G. PMID- 19807099 TI - Photodegradation of flumorph in aqueous solutions and natural water under abiotic conditions. AB - Flumorph is an oomycete fungicide that is now used extensively in China (Hu, J. Y.; Liu, C.; Yan, H. Degradation of flumorph in soils, aqueous buffer solutions, and natural waters. J. Agric. Food Chem. 2008, 56, 8574-8579). The photodegradation of flumorph in aqueous solutions and natural water have been assessed under natural and controlled conditions in this work. The kinetics of photodecomposition of flumorph was determined using the high-performance liquid chromatography (HPLC)-diode array detector (DAD), and the identification of photoproducts was carried out with HPLC-mass spectrometry (MS) [electrospray ionization (ESI) positive mode]. The rate of photodecomposition of flumorph in aqueous solutions and natural water followed first-order kinetics in both UV radiation and natural sunlight, and the Z isomer of flumorph could convert to the E isomer. The degradation rates were faster under UV light than sunlight, with the half-lives (t(1/2) = ln 2/k) of 36.5-64.2 min and 36.3-73.1 days, respectively. One major photoproduct was detected in UV light and tentatively identified according to HPLC-MS spectral information as (E or Z)-3-(3, 4 dimethoxyphenyl)-3-(4-fluorophenyl)-2-acrylamide. Photosensitizers, such as H(2)O(2) and riboflavin, could enhance photolysis of flumorph in natural sunlight. The results obtained indicated that photoreaction was an important dissipation pathway of flumorph in natural water systems. PMID- 19807100 TI - Rapid evaluation of oxidized fatty acid concentration in virgin olive oils using metal oxide semiconductor sensors and multiple linear regression. AB - This works aims to set up a rapid and nondestructive method to evaluate the advanced oxidation of virgin olive oils (VOOs). An electronic nose based on an array of six metal oxide semiconductor sensors was used, jointly with multiple linear regression (MLR), to predict the oxidized fatty acid (OFA) concentration in VOO samples characterized by different oxidative status. An MLR model constructed using five predictors was able to predict OFA concentration with an average validation error of 9%. PMID- 19807101 TI - Study of the migration of photoinitiators used in printed food-packaging materials into food simulants. AB - Photoinitiators (PIs) are used as catalyzers for inks and lacquers that are cured with ultraviolet (UV) light, and they can contaminate foodstuffs by mass transference.The aim of the present paper is to study the migration of six photoinitiators (Irgacure 184, benzophenone, Irgacure 651, Irgacure 907, ITX, and EHA) into food simulants [distilled water, 3% acetic acid (w/v) in aqueous solution, 10, 20, 30, 60 and 95% ethanol (v/v) in aqueous solution]. Migration levels of the six PIs into different food simulants were compared after a 30 day contact period with the additivated plastic. A relationship between R (ratio between log K(o/w) and MW) and total migration was found for PIs with log K(o/w) < 5. Key parameters of migration processes were calculated according to a mathematical model based on Fick's second law. Diffusion (D) and partition coefficients were estimated and compared among different simulants, temperatures, and PIs to understand better the mechanisms of the migration process and the physicochemical properties that most influence this phenomenon. For instance, at 5 degrees C, in ethanol 95% (v/v), D ranged between 4.2 x 10(-11) cm/s for Irgacure 907 and 3.0 x 10(-9) cm/s for benzophenone. PMID- 19807103 TI - 1,1'-Disubstituted ferrocenes as molecular hinges in mono- and bivalent dopamine receptor ligands. AB - On the basis of previous work on dopaminergic partial agonists of type 1 and 2, disubstituted ferrocenes are presented as valuable arene bioisosteres. Because substituents at the distal cyclopentadienyl ring are able to adjust the relative disposition that is required for ligand binding, disubstituted ferrocenes can act as molecular hinges. Taking advantage of click chemistry, the regioselective construction and functionalization of the target molecules is reported. Thus triazole derived appendages were used for the fine-tuning of biological activity and for the attachment of linker units generating bivalent GPCR ligands. Receptor binding was evaluated by radioligand displacement experiments, revealing superaffinity with sub- to single-digit nanomolar K(i) values for particular test compounds. As a neutral antagonist at the dopamine receptors D3 and D4 and a potent partial agonist at the D2 subtype (intrinsic activity = 57%, EC(50) = 2.5 nM), the bifunctional ferrocene 10b revealed a novel and unique activity profile. PMID- 19807104 TI - Cadmium and nickel uptake are differentially modulated by salicylic acid in Matricaria chamomilla plants. AB - Chamomile (Matricaria chamomilla) is a widely used medicinal plant which also accumulates heavy metals in its above-ground organs. We investigated the effect of the important plant signaling molecule, salicylic acid (SA), on the accumulation of Ni or Cd, by exposing plants over 7 days to 60 microM solutions of individual heavy metals with or without 50 microM SA. Special emphasis was focused on phenolic metabolism-related parameters, not only because of their importance for growth and stress tolerance but also because phenolics are potent antioxidants in human diet. In combined treatments, SA stimulated an increase in soluble proteins of roots and reduced their water content. SA reduced total Cd in the shoot and increased Ni. Total and "intraroot" Ni decreased in Ni + SA treatment, while in the case of Cd, only "intraroot" content decreased in Cd + SA treatment, being correlated with cell wall-bound phenolic acids and lignin. SA was strongly accumulated in roots from the Ni + SA treatment, being correlated with an increase in hydrogen peroxide. In both Cd + SA and Ni + SA treatments, SA enhanced phenylalanine ammonia-lyase activity and accumulation of total soluble phenols, particularly in the roots. Here, we report for the first time that soluble phenols may be involved in Cd shoot-to-root translocation. In the case of Ni, it seems that phenols serve as a root barrier in order to prevent Ni from reaching the above-ground organs. The effects of SA on phenolic metabolism, and the signaling role of ROS in the accumulation of phenols, are discussed. PMID- 19807105 TI - Detection and quantitation of 2,5-diketopiperazines in wheat sourdough and bread. AB - Liquid chromatography mass spectrometry (LC-MS) was used to quantify the levels of the 2,5-diketopiperazines cis-cyclo(L-Leu-L-Pro) and cis-cyclo(L-Phe-L-Pro) in acidified dough and bread. Dough acidification led to a significant increase in the level of cis-cyclo(L-Leu-L-Pro) and cis-cyclo(L-Phe-L-Pro) over 48 h compared to a nonacidified dough. However, no differences were found between chemically (mix of lactic and acetic acid in the presence of antibiotics) and biologically acidified doughs. On examination of the levels of cis-cyclo(L-Leu-L-Pro) and cis cyclo(L-Phe-L-Pro) in bread crumb and crust, it was found that temperature is the main causative agent of 2,5-diketopiperazine formation during the baking process. Bread crumb and crust contained almost 100 and 2000 times respectively the levels found in dough prior to baking. cis-Cyclo(L-Leu-L-Pro) and cis-cyclo(L-Phe-L-Pro) were also found to be at sensorally active levels in bread crust, however both 2,5-diketopiperazines were found to be below the minimum inhibitory concentration for antifungal activity in bread. PMID- 19807106 TI - 7-Hydroxy-benzopyran-4-one derivatives: a novel pharmacophore of peroxisome proliferator-activated receptor alpha and -gamma (PPARalpha and gamma) dual agonists. AB - Design, synthesis, and in vitro bioevaluation of a new class of potential dual PPARalpha and gamma agonists discovered through a structure-driven design paradigm are described. The 7-hydroxy-benzopyran-4-one moiety (includes flavones, flavanones, and isoflavones) is the key pharmacophore of these novel molecules, exhibiting similarity to the core structure of both fibrates and thiazolidinediones. New lead PPAR ligands were identified from "natraceuticals" and synthetic analogues. In total, 77 molecules, including chalcones, flavones, flavanones, isoflavones, and pyrazole derivatives, were screened and structure activity relationship studies of the dual agonists undertaken. Compounds 68, 70, 72, and 76 were identified as novel and potent dual PPARalpha and gamma agonists. These novel molecules may have the potential to be the future leads in PPAR related disorders, including type II diabetes mellitus and metabolic syndrome. PMID- 19807107 TI - Characterization of IgG N-glycans employing a microfluidic chip that integrates glycan cleavage, sample purification, LC separation, and MS detection. AB - A novel polymeric microfluidic device with an on-chip enzyme reactor has been developed for the characterization of recombinant glycoproteins. The enzyme reactor chip packed with PNGase F-modified solid support material was combined with a microfluidic glycan cleanup chip and a commercially available HPLC-chip to perform glycoprotein deglycosylation, protein removal, glycan capture, glycan LC separation, and nanoelectrospray into a time-of-flight mass spectrometry (TOF-MS) system. With this integrated chip, the combined sample preparation and sample analysis time was reduced from multiple hours to less than 10 min. A once tedious and time-consuming glycan analysis workflow is now integrated into an HPLC-chip device. Glycan profiling analysis has been achieved with as little as 100 ng of monoclonal antibody. Furthermore, a single chip was shown to retain activity and perform equivalently for over 250 replicate glycan profiles from a recombinant antibody. PMID- 19807108 TI - Shape-controlled synthesis of highly crystalline titania nanocrystals. AB - A versatile synthetic method based on solvothermal technique has been developed for the fabrication of TiO(2) nanocrystals with different shapes such as rhombic, truncated rhombic, spherical, dog-bone, truncated and elongated rhombic, and bar. The central features of our approach are the use of water vapor as hydrolysis agent to accelerate the reaction and the use of both oleic acid and oleylamine as two distinct capping surfactants which have different binding strengths to control the growth of the TiO(2) nanoparticles. We also show that the presence of an appropriate amount of water vapor along with the desired oleic acid/oleylamine molar ratio plays a crucial role in controlling size and shape of TiO(2) nanocrystals. PMID- 19807109 TI - Compound-specific delta34S analysis of volatile organics by coupled GC/multicollector-ICPMS. AB - We have developed a highly sensitive and robust method for the analysis of delta(34)S in individual organic compounds by coupled gas chromatography (GC) and multicollector inductively coupled plasma mass spectrometry (MC-ICPMS). The system requires minimal alteration of commercial hardware and is amenable to virtually all sample introduction methods. Isobaric interference from O(2)(+) is minimized by employing dry plasma conditions and is cleanly resolved at all masses using medium resolution on the Thermo Neptune MC-ICPMS. Correction for mass bias is accomplished using standard-sample bracketing with peaks of SF(6) reference gas. The precision of measured delta(34)S values approaches 0.1 per thousand for analytes containing >40 pmol S and is better than 0.5 per thousand for those containing as little as 6 pmol S. This is within a factor of 2 of theoretical shot-noise limits. External accuracy is better than 0.3 per thousand. Integrating only the center of chromatographic peaks, rather than the entire peak, offers significant gain in precision and chromatographic resolution with minimal effect on accuracy but requires further study for verification as a routine method. Coelution of organic compounds that do not contain S can cause degraded analytical precision. Analyses of crude oil samples show wide variability in delta(34)S and demonstrate the robustness and precision of the method in complex environmental samples. PMID- 19807111 TI - Combination atmospheric pressure solids analysis probe and desorption electrospray ionization mass spectrometry ion source. AB - The atmospheric solids probe analysis (ASAP) probe was investigated as a means for conducting desorption electrospray ionization (DESI) using a commercially available ion source. Solid and liquid samples as well as a raw urine sample were placed on individual melting point tubes and were inserted into either a heated gas stream for ASAP analysis or a charged solvent spray for DESI-like experiments. Samples ranged in polarity, volatility, mass, (100-17,000 Da), and concentration (neat to submicromolar). Mass spectra of multiple samples were obtained within a single acquisition on a short time scale (<30 s per sample). The configuration of the ion source also allowed rapid switching (approximately 1 min) between ASAP, electrospray ionization (ESI), and DESI. PMID- 19807110 TI - Abasic site-containing DNAzyme and aptamer for label-free fluorescent detection of Pb(2+) and adenosine with high sensitivity, selectivity, and tunable dynamic range. AB - An abasic site called dSpacer has been introduced into duplex regions of the 8-17 DNAzyme and adenosine aptamer for label-free fluorescent detection of Pb(2+) and adenosine, respectively. The dSpacer can bind an extrinsic fluorescent compound, 2-amino-5,6,7-trimethyl-1,8-naphthyridine (ATMND), and quench its fluorescence. Addition of Pb(2+) enables the DNAzyme to cleave its substrate and release ATMND from DNA duplex, recovering the fluorescence of ATMND. Similarly, the presence of adenosine induces structural switching of the aptamer, resulting in the release of ATMND from the DNA duplex and a subsequent fluorescence enhancement. Under optimized conditions, this label-free method exhibits detection limits of 4 nM for Pb(2+) and 3.4 muM for adenosine, which are even lower than those of the corresponding labeled-DNAzyme and aptamer sensors. These low detection limits have been obtained without compromising any of the selectivity of the sensors. Finally, the dynamic range of the adenosine sensor has been tuned by varying the number of hybridized base-pairs in the aptamer duplex. The method demonstrated here can be applied for label-free detection and quantification of a broad range of analytes using other DNAzymes and aptamers. PMID- 19807112 TI - Determination of the macromolecular dimensions of hydrophobically modified polymers by micellar size exclusion chromatography coupled with multiangle light scattering. AB - The present work demonstrates that the use of a nonionic surfactant in the mobile phase together with light scattering coupled to size exclusion chromatography (SEC) provides an accurate determination of macromolecular dimensions of hydrophobically modified water-soluble polymer and polyelectrolyte, i.e., weight average molar mass M(w) and polydispersity I(p). This method, called micellar SEC, is based on the dissociation of the aggregates in aqueous solution and the formation of mixed micelles between the surfactant and the polymer hydrophobic groups. The methodology and its application are presented for synthetic sulfonated polyacrylamides (5 and 20 mol %) modified with three hydrophobic alkyl side groups (C8, C12, and C18) and with Triton X-100 as a nonionic surfactant and are discussed according to the associativity of polymers. The results are compared to those obtained by classical SEC in 0.1 M NaNO(3) and by static light scattering in formamide solution. PMID- 19807113 TI - Immunoassay of goat antihuman immunoglobulin G antibody based on luminescence resonance energy transfer between near-infrared responsive NaYF4:Yb, Er upconversion fluorescent nanoparticles and gold nanoparticles. AB - Near-infrared (NIR) light can penetrate biological samples and even tissues without causing sample damage and avoid autofluorescence from biological samples in fluorescence detection. Thus, a luminescence resonance energy transfer (LRET) based immunoassay that can be excited by NIR irradiation is a promising approach to the analysis of biological samples. Here we demonstrate the use of NIR-to visible upconversion nanoparticles (UCNPs) as an energy donor, which can emit a visible light upon the NIR irradiation, and gold nanoparticles (Au NPs) as an energy acceptor, which can absorb the visible light emitted from the donor, to develop a sandwich-type LRET-based immunoassay for the detection of goat antihuman immunoglobulin G (IgG). Amino-functionalized NaYF(4):Yb, Er UCNPs and Au NPs were first prepared and then conjugated with the human IgG and rabbit antigoat IgG, respectively. The NIR-excited fluorescence emission band of human IgG-conjugated NaYF(4):Yb, Er UCNPs (lambda(max) = 542 nm) partially overlaps with the visible absorption band of the rabbit antigoat IgG-conjugated colloidal Au NPs (lambda(max) = 530 nm), satisfying the requirement of spectral overlap between donors and acceptors for LRET. A LRET system was then formed when goat antihuman IgG was added to a mixture of human IgG-modified NaYF(4):Yb, Er UCNPs (donor) and rabbit antigoat IgG-modified Au NPs (acceptor). The sandwich-type immunoreactions between the added goat antihuman IgG (primary antibody) and the two different proteins (antigen and secondary antibody on the surface of the donors and acceptors, respectively) cross-bridge the donors and acceptors and thus shorten their spacing, leading to the occurrence of LRET from UCNPs to Au NPs upon NIR irradiation. As a result, the quenching of the NIR-excited fluorescence of the UCNPs is linearly correlated to the concentration of the goat antihuman IgG (in the range of 3-67 microg x mL(-1)) present in the system, enabling the detection and quantification of the antibody. Such sandwich-type LRET-based approach can reach a very low detection limit of goat antihuman IgG (0.88 microg x mL(-1)), indicating that this method is applicable for the trace protein detection. This approach is expected to be extended to the detection of other biological molecules once the donor and acceptor nanoparticles are modified by proper molecules that can recognize the target biomolecules. PMID- 19807114 TI - Effect of Costaria costata fucoidan on expression of matrix metalloproteinase-1 promoter, mRNA, and protein. AB - Fucoidans are sulfated fucosylated polymers from brown algae cell walls. We assessed the inhibitory effects of Costaria costata fucoidan on UVB-induced MMP-1 promoter, mRNA, and protein expression in vitro using the immortalized human keratinocyte (HaCaT) cell line. Pretreatment with fucoidan significantly inhibited MMP-1 protein expression compared to UVB irradiation alone. Fucoidan significantly reduced MMP-1 mRNA expression and inhibited UVB-induced MMP-1 promoter activity by 37.3%, 53.3%, and 58.5% at 0.01, 0.1, and 1 microg/mL, respectively, compared to UVB irradiation alone. C. costata fucoidan may be a potential therapeutic agent to prevent and treat skin photoaging. PMID- 19807115 TI - Fluorous aryldiazirine photoaffinity labeling reagents. AB - Two fluorous versions of trifluoromethyldiazirine derivatives have been designed and synthesized. The new photoaffinity labeling reagents have reactivity similar to that of their aryltrifluoromethyldiazirine parent when activated in MeOH, while the reaction products can be efficiently separated over fluorous silica gel. The alcohol group in the two reagents is further converted to activated carboxylic acid and amine, which enable coupling both reagents with small molecules and macromolecules under mild conditions. PMID- 19807116 TI - Traceless synthesis of hydantoin fused tetrahydro-beta-carboline on ionic liquid support in green media. AB - A novel ionic liquid (IL) supported, green synthetic protocol has been developed toward the synthesis of oxo and thio hydantoin analogues tethered with tetrahydro beta-carboline by the use of focused microwave irradiation. IL-bound tryptophan underwent a Pictet-Spengler reaction with various carbonyl compounds to generate the IL-immobilized tetrahydro-beta-carbolines in aqueous isopropanol media. Subsequent reaction of substituted tetrahydro-beta-carboline derivatives with various isocyanates and isothiocyanate provided a three-dimensional combinatorial library in a traceless fashion. PMID- 19807117 TI - Dinuclear gold-silver resting states may explain silver effects in gold(I) catalysis. AB - The resting state of the gold(I)-catalyzed hydroarylation of 1 changes in the presence of Ag(+), with silver free catalysts resting at the dinuclear gold structure 5 and Ag(+) containing solutions resting at a heteronuclear species like 6. Adventitious Ag(+) (typically from LAuCl activation) can therefore intercept key organogold intermediates and effect the catalysis even when it does not effect the reaction in Au free control experiments. PMID- 19807118 TI - Simple view on fingering instability of debonding soft elastic adhesives. AB - We study the crack-front fingering instability of an elastic adhesive tape that is peeled off a solid substrate. Our analysis is based on an energy approach using fracture mechanics and scaling laws and provides simple physical explanations for (i) the fact that the wavelength depends only on the thickness of the adhesive film and (ii) the threshold of the instability, and (iii) additionally estimates the characteristic size of the fingers. The scaling laws for these three observables are in agreement with existing experimental data. PMID- 19807119 TI - Can Coulomb Sturmians be used as a basis for N-electron molecular calculations? AB - A method is proposed for using isoenergetic configurations formed from many center Coulomb Sturmians as a basis for calculations on N-electron molecules. Such configurations are solutions to an approximate N-electron Schrodinger equation with a weighted potential, and they are thus closely analogous to the Goscinskian configurations that we have used previously to study atomic spectra. We show that when the method is applied to diatomic molecules, all of the relevant integrals are pure functions of the parameter s = kR, and therefore they can be evaluated once and for all and stored. PMID- 19807120 TI - Ferrocenyl alkanethiols-thio beta-cyclodextrin mixed self-assembled monolayers: evidence of ferrocene electron shuttling through the beta-cyclodextrin cavity. AB - This paper reports the preparation and characterization of an Au electrode modified with self-assembled alkane ferrocenes, in the absence and in the presence of beta-cyclodextrins (betaCD). Electrode modification with ferrocene derivatives was achieved through a self-assembled monolayer (SAM) approach, using ferrocenyl hexane thiol (FcC6) and ferrocenyl undecane thiol (FcC11); the same was also done using per-6-thio-beta-cyclodextrin. The different SAMs prepared were characterized by both cyclic voltammetry and electrochemical surface plasmon resonance (EC-SPR). The behavior of both single and binary monolayers including their interfacial reorganization was investigated and critically discussed, according to the nature of the SAM used. Cyclic voltammetry combined with SPR measurements revealed the reorientation of the SAM concomitant with the oxidation of ferrocene moieties. In particular, the electron shuttling of FcC11 through the betaCD cavity (mixed SAM) was also evidenced by both SPR and the electrocatalytic oxidation of ferro(II)cyanide. PMID- 19807121 TI - Structural and electronic properties of amino acid based ionic liquids: a theoretical study. AB - The gas-phase ion pairs of the ionic liquids containing 1-ethyl-3 methylimidazolium ([emim](+)) and 20 natural amino acids ([AA](-)) are studied at the B3LYP/6-311+G (d,p) level. The optimized structures, energies, and natural population analysis are presented and analyzed in terms of their possible correlation with the interaction energies and the H-bond separations. It is found that all the ion pairs of [emim][AA] can form strong H-bond interactions, which are dominated by the side-chain structure and the functional group of amino acid anions. The calculations indicate that an increase of the alkyl side-chain length coincides with a gradual decrease of H-bond energy, while the functional groups lead to the different localized charges on the anions, consequently affecting the electrostatic force. In addition, the intramolecular H bond in [AA](-) can weaken the interaction, due to the decrease of the proton-accepting ability of the carbonyl O atoms. The H-bond chemical nature of [emim][AA] is investigated by atoms in molecules and natural bond orbital analyses. The preliminary analysis of 20 kinds of [emim][AA] ion pairs provides some initial hints as to the relationship between the interaction energy and the experimental glass transition temperature. PMID- 19807122 TI - DNA interstrand cross-link formation by the 1,4-dioxobutane abasic lesion. AB - The oxidized abasic lesion 5'-(2-phosphoryl-1,4-dioxobutane) (DOB) is produced concomitantly with a single-strand break by a variety of DNA-damaging agents that abstract a hydrogen atom from the C5'-position. Independent generation of the DOB lesion in DNA reveals that it reversibly forms interstrand cross-links (ICLs) selectively with a dA opposite the 3'-adjacent nucleotide. Product studies and the use of monoaldehyde models suggest that ICL formation involves condensation of the dialdehyde with the exocyclic amine. Mechanistic studies and inspection of molecular models indicate that the local DNA environment and proximity of the exocyclic amine determine the selectivity for reaction with dA. Proximity control of the electrophile's reactivity is distinct from that of structurally similar freely diffusing molecules. ICL formation by a DOB lesion that is adjacent to a single-strand break is potentially significant because the product constitutes a "clustered" or "complex" lesion. Clustered lesions can lead to highly deleterious double-strand breaks upon nucleotide excision repair. PMID- 19807123 TI - Synthesis of ZnO hexagonal single-crystal slices with predominant (0001) and (0001) facets by poly(ethylene glycol)-assisted chemical bath deposition. AB - ZnO hexagonal single-crystal slices with predominant (0001) and (0001) facets have been fabricated by poly(ethylene glycol)-assisted chemical bath deposition. PMID- 19807124 TI - Chemical library screens targeting an HIV-1 accessory factor/host cell kinase complex identify novel antiretroviral compounds. AB - Nef is an HIV-1 accessory protein essential for AIDS progression and an attractive target for drug discovery. Lack of a catalytic function makes Nef difficult to assay in chemical library screens. We developed a high-throughput screening assay for inhibitors of Nef function by coupling it to one of its host cell binding partners, the Src-family kinase Hck. Hck activation is dependent upon Nef in this assay, providing a direct readout of Nef activity in vitro. Using this screen, a unique diphenylfuropyrimidine was identified as a strong inhibitor of Nef-dependent Hck activation. This compound also exhibited remarkable antiretroviral effects, blocking Nef-dependent HIV replication in cell culture. Structurally related analogs were synthesized and shown to exhibit similar Nef-dependent antiviral activity, identifying the diphenylfuropyrimidine substructure as a new lead for antiretroviral drug development. This study demonstrates that coupling noncatalytic HIV accessory factors with host cell target proteins addressable by high-throughput assays may afford new avenues for the discovery of anti-HIV agents. PMID- 19807125 TI - In vitro bioaccessibility of beta-carotene from heat-processed orange-fleshed sweet potato. AB - Orange-fleshed sweet potato (OFSP) is currently promoted in parts of sub-Saharan Africa as a biofortified staple food with large potential to provide considerable amounts of provitamin A carotenoids. However, the bioaccessibility of provitamin A carotenoids from OFSP has not been widely investigated, especially not as an effect of different preparation methods. In this study, we used an in vitro digestion model to assess the bioaccessibility of beta-carotene from differently heat-processed OFSP. The fraction of carotenoids transferred from the food matrix to a micellar phase obtained after microfiltration and to a supernatant obtained after low-speed centrifugation was investigated. The percentage of accessible all trans-beta-carotene in the micellar phase varied between 0.5 and 1.1% in the heat processed OFSP without fat and between 11 and 22% with the addition of 2.5% (w/w) cooking oil. In comparison with the micellar phase, the percentage of accessible all-trans-beta-carotene in the supernatant phase was significantly higher (P < 0.001), between 24 and 41% without fat and between 28 and 46% with fat. These results support the importance of fat for an improved micellarization of beta carotene. Overall, the high in vitro bioaccessibility of beta-carotene from heat processed OFSP indicates that sweet potato might be a promising dietary approach to combat vitamin A deficiency. PMID- 19807126 TI - Activation and inhibition of nuclear factor kappa B activity by cereal extracts: role of dietary phenolic acids. AB - The transcription factor nuclear factor kappa B (NF-kappaB) plays a critical role in stress, immune, and inflammatory responses, and the modulation of its activity can be a potentially effective preventive strategy for controlling certain diseases. Cereal grains contain phenolic compounds in concentrations comparable to those in fruits and vegetables, well-known for their beneficial effect on human health. In this study we aimed to examine the effect of different phenolic extracts from barley, oat, wheat, and buckwheat on the modulation of basal and lipopolysaccharide (LPS)-induced NF-kappaB activity and elucidate the role of phenolic acids in this modulation. Three extracts were prepared: extracts of free phenolic compounds (M1), extracts of free phenolic acids (M2), and extracts of bound phenolic acids (HY). Generally, extracts M2 showed the highest effect on modulation of NF-kappaB activity with strong inhibition of LPS-induced NF-kappaB activity at all concentrations and of the basal NF-kappaB activity at concentrations equal to or lower than 3 mg/mL. Most of extracts M1 and HY slightly increased both the basal and the LPS-induced NF-kappaB activation. However, at the highest concentrations (3 or 15 mg/mL) extracts HY inhibited LPS induced NF-kappaB activation. Similar experiments with standard solutions of phenolic acids indicated their ability to modulate the NF-kappaB activity. PMID- 19807127 TI - Effect of lipophilization of hydroxytyrosol on its antioxidant activity in fish oils and fish oil-in-water emulsions. AB - The effect of lipophilization of the antioxidant efficiency of hydroxytyrosol on fish oil enriched systems was studied. Hydroxytyrosol fatty acid esters with increasing size of the alkyl chain and different lipophilicity were tested in bulk fish oils and fish oil-in-water emulsions. Results showed a significant antioxidant activity of hydroxytyrosol esters in both systems especially in emulsions. The introduction of a lipophilic chain decreased the antioxidant effectiveness of hydroxytyrosol in homogeneous systems as fish oils. In emulsion systems, the presence of a short-medium lipophilic chain (acetate, butyrate or octanoate) improved the antioxidant efficiency of hydroxytyrosol favoring the physical location of the antioxidant in the interface, but longer alkyl chain (laurate) maintained or even decreased their antioxidant activity. A maximum of antioxidant efficiency seems to appear when the chain length of the hydroxytyrosol derivative is that of eight carbons which is probably associated with a preferential location of the diorthophenolic moiety in the right geometry. These results are of high importance for the optimum design of effective antioxidants for omega 3 enriched foods, which are very susceptible to suffer oxidation and, then, rancidity. PMID- 19807128 TI - Mapping the phosphorylation sites of Ulk1. AB - Ulk1 is a serine/threonine kinase that controls macroautophagy, an essential homeostatic recycling pathway that degrades bulk cytoplasmic material and directs the turnover of organelles such as peroxisomes and mitochondria. Further, macroautophagy is potently induced by signals that trigger metabolic stress, such as hypoxia and amino acid starvation, where its recycling functions provide macromolecules necessary to maintain catabolic metabolism and cell survival. Substrates for Ulk1 have not been identified, and little is known regarding post translational control of Ulk1 kinase activity and function. To gain insights into the regulatory mechanisms of Ulk1, we developed a robust purification protocol for Ulk1 and demonstrated that Ulk1 is highly phosphorylated and requires autophosphorylation for stability. Importantly, high-resolution, tandem mass spectrometry identified multiple sites of phosphorylation on Ulk1, including several within domains known to regulate macroautophagy. Differential phosphorylation analyses also identified sites of phosphorylation in the C terminal domain that depend upon or require Ulk1 autophosphorylation. PMID- 19807129 TI - Single nanorod devices for battery diagnostics: a case study on LiMn2O4. AB - This paper presents single nanostructure devices as a powerful new diagnostic tool for batteries with LiMn(2)O(4) nanorod materials as an example. LiMn(2)O(4) and Al-doped LiMn(2)O(4) nanorods were synthesized by a two-step method that combines hydrothermal synthesis of beta-MnO(2) nanorods and a solid state reaction to convert them to LiMn(2)O(4) nanorods. lambda-MnO(2) nanorods were also prepared by acid treatment of LiMn(2)O(4) nanorods. The effect of electrolyte etching on these LiMn(2)O(4)-related nanorods is investigated by both SEM and single-nanorod transport measurement, and this is the first time that the transport properties of this material have been studied at the level of an individual single-crystalline particle. Experiments show that Al dopants reduce the dissolution of Mn(3+) ions significantly and make the LiAl(0.1)Mn(1.9)O(4) nanorods much more stable than LiMn(2)O(4) against electrolyte etching, which is reflected by the magnification of both size shrinkage and conductance decrease. These results correlate well with the better cycling performance of Al-doped LiMn(2)O(4) in our Li-ion battery tests: LiAl(0.1)Mn(1.9)O(4) nanorods achieve 96% capacity retention after 100 cycles at 1C rate at room temperature, and 80% at 60 degrees C, whereas LiMn(2)O(4) shows worse retention of 91% at room temperature, and 69% at 60 degrees C. Moreover, temperature-dependent I-V measurements indicate that the sharp electronic resistance increase due to charge ordering transition at 290 K does not appear in our LiMn(2)O(4) nanorod samples, suggesting good battery performance at low temperature. PMID- 19807130 TI - Electrically conductive and optically active porous silicon nanowires. AB - We report the synthesis of vertical silicon nanowire array through a two-step metal-assisted chemical etching of highly doped n-type silicon (100) wafers in a solution of hydrofluoric acid and hydrogen peroxide. The morphology of the as grown silicon nanowires is tunable from solid nonporous nanowires, nonporous/nanoporous core/shell nanowires, to entirely nanoporous nanowires by controlling the hydrogen peroxide concentration in the etching solution. The porous silicon nanowires retain the single crystalline structure and crystallographic orientation of the starting silicon wafer and are electrically conductive and optically active with visible photoluminescence. The combination of electronic and optical properties in the porous silicon nanowires may provide a platform for novel optoelectronic devices for energy harvesting, conversion, and biosensing. PMID- 19807131 TI - Raman spectroscopy of ripple formation in suspended graphene. AB - Using Raman spectroscopy, we measure the optical phonon energies of suspended graphene before, during, and after thermal cycling between 300 and 700 K. After cycling, we observe large upshifts ( approximately 25 cm(-1)) of the G band frequency in the graphene on the substrate region due to compression induced by the thermal contraction of the underlying substrate, while the G band in the suspended region remains unchanged. From these large upshifts, we estimate the compression in the substrate region to be approximately 0.4%. The large mismatch in compression between the substrate and suspended regions causes a rippling of the suspended graphene, which compensates for the change in lattice constant due to the compression. The amplitude (A) and wavelength (lambda) of the ripples, as measured by atomic force microscopy, correspond to an effective change in length Deltal/l that is consistent with the compression values determined from the Raman data. PMID- 19807132 TI - H2-driven deoxygenation of epoxides and diols to alkenes catalyzed by methyltrioxorhenium. AB - Catalytic deoxygenation of epoxides and diols is underdeveloped. This reaction is appealing in the context of making value-added organics from biomass. Methyltrioxorhenium (MTO) catalyzes the conversion of epoxides and vicinal diols to olefins with dihydrogen (H(2)) as the reductant under reasonably mild conditions (150 degrees C and 80-300 psi). The only reaction byproduct is water. The reaction is selective for cis cyclic diols, signaling a mechanism of alkene extrusion from a coordinated epoxide via a metallaoxetane intermediate. PMID- 19807133 TI - Coordination-insertion copolymerization of fundamental polar monomers. PMID- 19807134 TI - Secbase: database module to retrieve secondary structure elements with ligand binding motifs. AB - Secbase is presented as a novel extension module of Relibase. It integrates the information about secondary structure elements into the retrieval facilities of Relibase. The data are accessible via the extended Relibase user interface, and integrated retrieval queries can be addressed using an extended version of Reliscript. The primary information about alpha-helices and beta-sheets is used as provided by the PDB. Furthermore, a uniform classification of all turn families, based on recent clustering methods, and a new helix assignment that is based on this turn classification has been included. Algorithms to analyze the geometric features of helices and beta-strands were also implemented. To demonstrate the performance of the Secbase implementation, some application examples are given. They provide new insights into the involvement of secondary structure elements in ligand binding. A survey of water molecules detected next to the N-terminus of helices is analyzed to show their involvement in ligand binding. Additionally, the parallel oriented NH groups at the alpha-helix N termini provide special binding motifs to bind particular ligand functional groups with two adjacent oxygen atoms, e.g., as found in negatively charged carboxylate or phosphate groups, respectively. The present study also shows that the specific structure of the first turn of alpha-helices provides a suitable explanation for stabilizing charged structures. The magnitude of the overall helix macrodipole seems to have no or only a minor influence on binding. Furthermore, an overview of the involvement of secondary structure elements with the recognition of some important endogenous ligands such as cofactors shows some distinct preference for particular binding motifs and amino acids. PMID- 19807135 TI - Nonracemic bicyclic lactam lactones via regio- and cis-diastereocontrolled C-H insertion. Asymmetric synthesis of (8S,8aS)-octahydroindolizidin-8-ol and (1S,8aS)-octahydroindolizidin-1-ol. AB - The Rh(2)(MPPIM)(4)-catalyzed intramolecular C-H insertion reaction of (S)- and (R)-1-benzyl-5-(alpha-diazoacetoxy)piperidin-2-one and (S)-1-benzyl-4-(alpha diazoacetoxy)pyrrolidin-2-one proceeds with high regioselectivity and cis diastereoselectivity to give good yields of chiral nonracemic bicyclic lactam lactones (BLLs). For (S)- and (R)-1-benzyl-5-(alpha-diazoacetoxy)piperidin-2-one, the regioselectivity of the C-H insertion is catalyst-dependent; for example, (S) 1-benzyl-5-(alpha-diazoacetoxy)piperidin-2-one undergoes C-H insertion at C-6 preferentially when Rh(2)(5S-MPPIM)(4) is used, but with Rh(2)(5R-MPPIM)(4), C-H insertion occurs preferentially at C-4. This effect is not observed in the reaction of (S)-1-benzyl-4-(alpha-diazoacetoxy)pyrrolidin-2-one. The utility of the BLLs as chiral building blocks in alkaloid synthesis is exemplified by the synthesis of (8S,8aS)-octahydroindolizidin-8-ol and (1S,8aS)-octahydroindolizidin 1-ol. PMID- 19807136 TI - Equilibrium acidities and homolytic bond dissociation enthalpies of the acidic C H bonds in phosphonates and related phosphorus containing compounds. AB - The homolytic bond dissociation enthalpies (BDEs) of the acidic C-H bonds in a series of phosphonates and related phosphorus-containing compounds have been determined by a combination (eq 3) of their equilibrium acidities (pK(HA)'s) and the oxidation potentials [E(ox)(A(-))'s] of their conjugate anions, which were measured in DMSO solution. The equilibrium acidity increases by 15.5, 14.9, and 10.9 pK units for the introduction of an alpha-P(O)(OEt)(2) group into toluene, acetonitrile, and ethyl acetate, respectively. Comparison with the published equilibrium acidity increases by 25.6, 24.4, and 21.0 pK units for the introduction of an alpha-(+)PPh(3) group into the same series of substrates indicates that the phosphoryl carbanions (Horner-Wadsworth-Emmons reagents) are about 10 pK units more reactive than the corresponding triphenylphosphonium ylides (Wittig reagents). However, both alpha-P(O)(OEt)(2) and alpha-(+)PPh(3) groups have negligible effects on the adjacent C-H BDEs, indicating that there is no resonance delocalization into the 3d vacant orbitals of phosphorus and that their acidifying effects are exclusively associated with the field/inductive (electrostatic) and polarizability effects. The acidifying effect of an alpha CO(2)Et group is shown to be ca. 3.5 pK units stronger than that of an alpha P(O)(OEt)(2) group in the same substrates, suggesting that the larger acidifying effect of the alpha-CO(2)Et group is predominantly associated with the resonance delocalization rather than with the field/inductive (electrostatic) and polarizability effects. PMID- 19807137 TI - Intensive chiroptical properties of chiral polyfluorenes associated with fibril formation. AB - Thin films of chiral poly{9,9-bis[(3S)-3,7-dimethyloctyl]-2,7-fluorene} (1) were studied using circular dichroism (CD) spectroscopy. Films spin coated from chloroform solution, show CD with a degree of polarization g(abs) (= +4 x 10(-4) at 400 nm) that is independent of film thickness (50-290 nm). This implies that g(abs) is an intensive property of the material and related to the chiral organization of the molecules on a length scale less than 50 nm. Atomic force microscopy (AFM) on the films reveals fibrils. Addition of nonsolvent methanol to a solution of 1 in chloroform leads to fibril formation in solution and results in CD similar in band shape to that of the pristine spin coated films from chloroform solution and a g(abs) comparable in magnitude. Thus the chiral molecular arrangement leading to circular dichroism is part of the internal structure of these fibrils. PMID- 19807138 TI - Microstructure, luminescence, and stability of a europium complex covalently bonded to an attapulgite clay. AB - Attapulgite clay, natural silicate nanorods, can form nanocomposites with extreme chemical stability and remarkable exposure durability. Combining luminescent lanthanide complexes with attapulgite to improve their stability and even luminescent properties is fascinating, promising but challenging in the lanthanide composite field. A europium complex Eu(tta)(3)(H(2)O)(2) (Htta = 2 thenoyltrifluoroacetone) was covalently coupled on attapulgite (and MCM-41 or ZSM 5 for comparison) via ligand exchange reaction, generating the first example of attapulgite-based ternary europium complexes. The composites were characterized by (29)Si magic-angle spinning (MAS) NMR, CHN elemental analysis, inductively coupled plasma-atomic emission spectroscopy (ICP) for Eu(3+) contents, X-ray diffraction (XRD), and UV-vis absorption spectra. The results indicate that Eu(3+) complexes bond covalently to the outer surfaces of attapulgite, permeate the channels, and are stuck with the complexes bonded to the inner walls of the pores in MCM-41, or invade into the channels of ZSM-5 after decomposition. These structures were further evidenced by luminescence efficiency and coordinated waters of the complexes linked to matrixes. The composites display more efficient emission, enhanced thermal stability, and improved exposure durability in comparison with the isolated complexes, due to interactions of the complexes with the matrixes. The most efficient emission of attapulgite-based complexes among the composites results from the uniformly structured ternary europium complexes. PMID- 19807140 TI - Structural dependence on excitation energy migration processes in artificial light harvesting cyclic zinc(II) porphyrin arrays. AB - A series of covalently linked cyclic porphyrin arrays CNZ that consist of N/2 of meso-meso directly linked zinc(II) porphyrin dimer subunits Z2 bridged by 1,3 phenylene spacers have been prepared by Ag(I)-promoted oxidative coupling reaction. We have investigated the excitation energy migration processes of CNZ in toluene by using femtosecond transient absorption anisotropy decay measurements by taking 2Z2 composed of two Z2 units linked by 1,3-phenylene as a reference molecule. On the basis of the excitation energy transfer rate determined for 2Z2, we have revealed the excitation energy hopping rates in the cyclic arrays CNZ by using a regular polygon model. The number of excitation energy hopping sites N(flat) calculated by using a regular polygon model is close to the observed N(expt) value obtained from the transient absorption anisotropy decays for C12Z-C18Z with circular and well-ordered structures. On the other hand, a large discrepancy between N(flat) and N(expt) was found for smaller or larger arrays (C10Z, C24Z, and C32Z). In the case of C10Z, m-phenylene linked 2Z2 motif with the interchromophoric angle of 120 degrees is not well suited to make a cyclic pentagonal array C10Z based on planar pentagonal structure. This geometrical factor inevitably causes a structural distortion in C10Z, leading to a discrepancy between N(expt) and N(flat) values. On the contrary, the presence of highly distorted conformers such as figure-eight structures reduces the number of effective hopping sites N(expt) in large cyclic arrays C24Z and C32Z. Thus, our study demonstrates that not only the large number of porphyrin chromophores in the cyclic arrays CNZ but the overall rigidity and three-dimensional orientation in molecular architectures is a key factor to be considered in the preparation of artificial light harvesting porphyrin arrays. PMID- 19807141 TI - Mass-dependent and mass-independent isotope effects of zinc in a redox reaction. AB - We report the isotope fractionation of zinc (Zn) associated with a redox reaction between Zn(0) and Zn(II). Zn isotopes were found fractionated in pyrometallurgical biphase extraction between liquid zinc and molten chloride. The isotopic composition of Zn in the molten chloride phase was analyzed by multiple collector inductively coupled plasma mass spectrometry and reported as (m)Zn/64Zn (m = 66, 67, and 68) ratios. The observed isotope fractionation consists of the mass-dependent and mass-independent isotope effects. The contributions of the nuclear mass and the nuclear volume to the overall isotope effect were evaluated by employing first-principles quantum calculations and using reported isotope shifts in atomic spectra. The magnitude of the mass-dependent isotope effect was explained by the sum of the isotope effect via intramolecular vibrations and the correction to the Born-Oppenheimer electronic energy. The mass-independent isotope effect was correlated with the Gibbs free energy change in the redox reaction. PMID- 19807142 TI - Dynamics of the O + CN reaction and N + CO scattering on two coupled surfaces. AB - Spin-orbit coupling between the two collinear (2)Pi and (4)Sigma(-) potential energy surfaces for the NCO system are calculated using the RASSI method with CASSCF wave functions as basis set. The GDVR method has been used to interpolate a spin-orbit coupling surface. Wave packet and quasi-classical trajectory surface hopping calculations have been performed and compared for both the O((3)P) + CN(X(2)Sigma(+)) --> N((4)S) + CO(X(1)Sigma(+)) reaction and for electronically inelastic scattering in the N + CO channels. The O + CN nonadiabatic reaction probabilities are small. The wavepacket study gives a resonance structure. Also for the N + CO electronically inelastic scattering the wave packet calculations give a distinct resonance structure with peak transition probabilities up to around 10%, which is somewhat lower than the trajectory surface hopping results. PMID- 19807143 TI - Beta-amino alcohol derived beta-hydroxy- and beta-(o diphenylphosphino)benzoyloxy(o-diphenylphosphino)benzamides: an ester-amide ligand structural model for the palladium-catalyzed allylic alkylation reaction. AB - A commercially available collection of beta-amino alcohols have been converted to their corresponding beta-hydroxy- and beta-(o-diphenylphosphino)benzoyloxy(o diphenylphosphino)benzamides 11a-f and 12a-f and have been employed in the Tsuji Trost asymmetric alkylation reaction with 1,3-diphenylpropenyl acetate. With the exception of ligands 11b and 11f, the beta-hydroxybenzoyloxy(o diphenylphosphino)benzamide ligands 11a-f primarily afforded the (R)-enantiomer of the product. In contrast, the bis(phosphine) ligands 12a-f consistently afforded the (S)-enantiomer. The best ligand (12c) was derived from cis-(1R,2S)-2 amino-1,2-diphenyl-1-ethanol, and when applied in the asymmetric allylic alkylation reaction, it yielded the product in an enantiomeric ratio of 97.8.22 favoring the (S)-enantiomer. A computational study was conducted on the conformation that this ligand might adopt in the palladium-catalyzed alkylation reaction as compared to that of the Trost ligand 1a. PMID- 19807144 TI - Conformational and structural determination of F(2)NC(O)F and F(2)NC(O)NCO. A joint experimental and theoretical study. AB - The vibrational properties of two difluoroaminocarbonyl species were analyzed by recording the FTIR spectra of the vapor for difluoroaminocarbonyl fluoride, F(2)NC(O)F, and difluoroaminocarbonyl isocyanate, F(2)NC(O)NCO. Moreover, the Raman spectrum of liquid F(2)NC(O)NCO was obtained. Vibrational assignments were made on the basis of a normal coordinate analysis and the evaluation of the band contours appearing in the FTIR spectrum of the vapor. The conformational space of both difluoroaminocarbonyl derivatives were studied by using the B3LYP and MP2 level of theory with extended basis sets [6-311+G(3df) and aug-cc-pVTZ]. Only one conformation belonging to the C(1) symmetry point group is expected for F(2)NC(O)F, whereas the overall evaluation of experimental and theoretical results suggests the existence of a mixture of two conformers for F(2)NC(O)NCO at room temperature. Its relative abundance of the most stable syn form (C=O double bond syn with respect to the N=C=O group) was estimated to be 56(5) %. PMID- 19807145 TI - Enzymatic one-pot route to telechelic polypentadecalactone epoxide: synthesis, UV curing, and characterization. AB - In an enzymatic one-pot procedure immobilized lipase B from Candida antarctica was used to synthesize semicrystalline diepoxy functional macromonomers based on glycidol, pentadecalactone, and adipic acid. By changing the stoichiometry of the building blocks, macromonomers of controlled molecular weight from 1400 to 2700 g mol(-1) could be afforded. The enzyme-catalyzed reaction went to completion (conversion >or=95%) within 24 h at 60 degrees C. After removal of the enzyme, the produced macromonomers were used for photopolymerization without any purification. The macromonomers readily copolymerized cationically with a cycloaliphatic diepoxide (Cyracure UVR-6110; CA-dE) to high conversion. The cross linked copolymers formed a durable film with a degree of crystallinity depending on the macromonomer size and amount of CA-dE used, without CA-dE the macromonomers homopolymerized only to a low degree. Combined with CA-dE conversions of 85-90% were determined by FT-Raman spectroscopy. The films became more durable once reinforced with CA-dE, increasing the cross-link density and reducing the crystallinity of the PDL segments in the films. PMID- 19807146 TI - Dynamics of formation of products D2CN+, DCN+, and CD3+ in the reaction of N+ with CD4: a crossed-beam and theoretical study. AB - The formation of D(2)CN(+) in the reaction of N(+) ((3)P) with CD(4) was studied using the crossed beam technique at collision energies of 3.66 and 4.86 eV. The experiments were complemented by calculations of stationary points on the triplet hypersurface of the system. The scattering data showed that the reaction proceeds by the formation of two intermediate complexes having different lifetimes: a long lived statistical intermediate and a short-lived complex (mean lifetime about one period of an average rotation) with more energy in translation than the statistical complex. Comparison with theoretical calculations suggests that the long-lived complex leads the CDND(+) isomer of the product ion, whereas the short lived complex leads prevailingly to the CD(2)N(+) isomer. The product DCN(+) results from further decomposition of the primary product D(2)CN(+), whereas CD(3)(+) is formed both by a hydride-ion transfer and a long-lived complex decomposition. PMID- 19807147 TI - Gas-phase Bronsted superacidity of some derivatives of monocarba-closo-borates: a computational study. AB - The structures and gas-phase acidities (GA) of several CB(11)H(12)H-based carborane acid derivatives (HA) have been calculated with DFT B3LYP method using 6-311+G**, 6-311++G** basis sets. In order to verify the obtained GA values, several systems were also studied at G3(MP2) level of theory. Inserted substituents (CF(3), F, Cl, Br, I, CN, CH(3), etc.) followed the "belts" of the monocarborane cage starting from the boron antipodal to the carbon. In general, the predicted intrinsic gas-phase acidities of the systems varied according to the substituents in the following order of decreasing strength: CF(3) > F > Cl > Br > I > CN > CH(3). Nevertheless, some inconsistencies occurred. F and CN derivatives with lower degree of substitution had weaker intrinsic acidities than the respective Cl derivatives, but the situation was reversed in the case of a larger number of substituents. To obtain better understanding how the substituents influence the basicity of the carborane anion, three hypothetical reaction series were investigated, in which the protonation center was fixed on the boron atom (B(12)), antipodal to the carbon (C(1)), and a single substituent replaced the hydrogens at the vertexes of the three remaining positions (C(1), B(2), and B(7)). The intrinsic gas-phase acidities in these series of neutral carborane-based acids CB(11)X(1)H(11)H are found to clearly depend on the field inductive and resonance effects of the substituent X. Some influence of the polarizability of X on the reaction center (B(12)) could be detected only in the alpha position (B(7)). PMID- 19807148 TI - Theory of proton-coupled electron transfer in energy conversion processes. AB - Proton-coupled electron transfer (PCET) reactions play an essential role in a broad range of energy conversion processes, including photosynthesis and respiration. These reactions also form the basis of many types of solar fuel cells and electrochemical devices. Recent advances in the theory of PCET enable the prediction of the impact of system properties on the reaction rates. These predictions may guide the design of more efficient catalysts for energy production, including those based on artificial photosynthesis and solar energy conversion. This Account summarizes the theoretically predicted dependence of PCET rates on system properties and illustrates potential approaches for tuning the reaction rates in chemical systems. A general theoretical formulation for PCET reactions has been developed over the past decade. In this theory, PCET reactions are described in terms of nonadiabatic transitions between the reactant and product electron-proton vibronic states. A series of nonadiabatic rate constant expressions for both homogeneous and electrochemical PCET reactions have been derived in various well-defined limits. Recently this theory has been extended to include the effects of solvent dynamics and to describe ultrafast interfacial PCET. Analysis of the rate constant expressions provides insight into the underlying physical principles of PCET and enables the prediction of the dependence of the rates on the physical properties of the system. Moreover, the kinetic isotope effect, which is the ratio of the rates for hydrogen and deuterium, provides a useful mechanistic probe. Typically the PCET rate will increase as the electronic coupling and temperature increase and as the total reorganization energy and equilibrium proton donor-acceptor distance decrease. The rate constant is predicted to increase as the driving force becomes more negative, rather than exhibit turnover behavior in the inverted region, because excited vibronic product states associated with low free energy barriers and relatively large vibronic couplings become accessible. The physical basis for the experimentally observed pH dependence of PCET reactions has been debated in the literature. When the proton acceptor is a buffer species, the pH dependence may arise from the protonation equilibrium of the buffer. It could also arise from kinetic complexity of competing concerted and sequential PCET reaction pathways. In electrochemical PCET, the heterogeneous rate constants and current densities depend strongly on the overpotential. The change in equilibrium proton donor acceptor distance upon electron transfer may lead to asymmetries in the Tafel plots and deviations of the transfer coefficient from the standard value of one half at zero overpotential. Applications of this theory to experimentally studied systems illustrate approaches that can be utilized to tune the PCET rate. For example, the rate can be tuned by changing the pH or using different buffer species as proton acceptors. The rate can also be tuned with site-specific mutagenesis in biological systems or chemical modifications that vary the substituents on the redox species in chemical systems. Understanding the impact of these changes on the PCET rate may assist experimental efforts to enhance energy conversion processes. PMID- 19807149 TI - Effect of different alkyl groups at the N-position on the luminescence of carbazole-based beta-diketonate europium(III) complexes. AB - A series of carbazole-based beta-diketone derivatives and their europium(III) ternary complexes Eu(N-Cx)(3)phen were designed and synthesized, where N-Cx denotes carbazole-based beta-diketonates with different alkyl substituents at N position of the carbazole ring and phen is 1,10-phenethroline. Thermogravimetric analysis (TGA) shows that the decomposition temperature of the complexes is over 360 degrees C. UV-visible absorption spectroscopy, photoluminescence (PL), and the luminescence quantum yield of the Eu(III) complexes were measured and compared with each other, and the effect of different substituted-alkyls at N position in the carbazole ring on the photoluminescence was discussed in details, indicating there exists a competition between the absorption capacity and the energy transfer efficiency for the complexes when the structure of the substituted-alkyls changes. The triplet state energy levels of the beta diketonate ligands in the complexes are higher than that of the lowest excited level of Eu(3+) ion, (5)D(0), so the photoluminescence mechanism of the Eu(III) complexes was proposed as a ligand-sensitized luminescence process. Red LEDs were fabricated by precoating the complexes onto 395 nm emitting InGaN chips. All the results show that this series of Eu(III) complexes is a promising candidate as a red component in fabrication of NUV-based white LEDs. PMID- 19807150 TI - Biologically active sequences in the mouse laminin alpha3 chain G domain. AB - The laminin alpha3 chain is mainly expressed at the skin, and its C-terminal G domain has a critical role in multiple biological functions. We screened for biologically active sites on the mouse laminin alpha3 chain G domain using 107 synthetic peptides on coated plates and conjugated to Sepharose beads with HT1080 human fibrosarcoma cells, HaCaT human skin keratinocyte cells, and human dermal fibroblasts (HDFs). Eleven peptides exhibited cell attachment activity with respect to the peptide-coated plates and/or peptide-Sepharose beads. MA3G28 (WTIQTTVDRGLL) strongly binds to HaCaT cells. Four peptides promoted PC12 cell neurite outgrowth. Heparin inhibited attachment of HDFs to eight peptides on the coated plates. In contrast, EDTA significantly inhibited attachment of HDFs to MA3G27 (NAPFPKLSWTIQ) and MA3G28 but had no effect on the attachment of the other peptides. HDF cells formed well-organized actin stress fibers and focal contacts with vinculin accumulation on MA3G27. Additionally, attachment of HDFs to MA3G27 was inhibited by anti-alpha6 and anti-beta1 integrin antibodies, suggesting that MA3G27 promotes alpha6beta1 integrin-mediated cell adhesion. MA3G57 (NQRLASFSNAQQS) exhibited cell attachment activity only in the peptide bead assay. MA3G57 conjugated to a chitosan membrane promoted HDF attachment and spreading with well-organized actin stress fibers. The anti-beta1 integrin antibody partially inhibited attachment of HDFs to the MA3G57-chitosan membrane, suggesting that the MA3G57 site is involved in beta1 integrin-mediated cell attachment. These active sites are likely important in the biological activities of the laminin alpha3 chain G domain and would be useful for the study of molecular mechanisms of laminin-receptor interactions. PMID- 19807151 TI - Determination of four carbamate pesticides in corn by cloud point extraction and high-performance liquid chromatography in the visible region based on their derivatization reaction. AB - A highly sensitive method for the determination of arprocarb (AC), carbofuran (CF), isoprocarb (IC), and fenobucarb (FC) is proposed. The method is based on alkaline hydrolysis of the four carbamate pesticides, and the resultant hydrolysis products are reacted with 4-aminoantipyrene (AP) to give four red color products. The colored compounds are enriched and separated by cloud point extraction (CPE) method, and the coacervate phase containing the compounds is determined with a high-performance liquid chromatography (HPLC) system in the visible region. AC, CF, IC, and FC were determined on the basis of a linear correlation between the signals of the colored compounds and the concentrations of the pesticides. The method is applied to determine the four pesticides in corn samples; the limits of detection are 2.0 x 10(-4) mg L(-1) for AC, CF, and IC and 5.0 x 10(-4) mg L(-1) for FC, with recoveries ranging between 84.8 and 93.0%, at spiking levels of 5 x 10(-3), 2 x 10(-2), and 0.2 mg kg(-1), respectively. PMID- 19807152 TI - Bioavailability and antioxidant effects of a xanthone-rich Mangosteen (Garcinia mangostana) product in humans. AB - Oxidative damage is involved in many chronic diseases including those cited as the major causes of death in Western societies such as cardiovascular disorders and cancer. Antioxidants may prevent these degenerative processes by various mechanisms including the scavenging of free radicals. Intake of antioxidant supplements is associated with preventing oxidative damages. This study investigated the absorption and antioxidant effects of a xanthone-rich mangosteen liquid in healthy human volunteers after the acute consumption of 59 mL of the supplement. The liquid contained mangosteen, aloe vera, green tea, and multivitamins. Results indicated that alpha-mangostin and vitamins B(2) and B(5) were bioavailable, with observed C(max) at t(max) of around 1 h. The antioxidant capacity measured with the oxygen radical absorbance capacity (ORAC) assay was increased with a maximum effect of 18% after 2 h, and the increased antioxidant level lasted at least 4 h. Overall, this study demonstrated the bioavailability of antioxidants from a xanthone-rich mangosteen product and its in vivo antioxidant effects. PMID- 19807153 TI - Detection of cyanidin in different-colored peanut testae and identification of peanut cyanidin 3-sambubioside. AB - Peanut testae are potent sources of polyphenols. When the water extracts and acid hydrolysates of five different-colored testae were analyzed by HPLC, chromatograms monitored at 280 nm varied remarkably, whereas two major peaks in the chromatograms monitored at 530 nm were detected only in kernels having completely or partially black color. After acid hydrolysis of the extracts, cyanidin was detected in each of the hydrolysates. By respectively subjecting the black testae of raw and roasted (175 degrees C for 20 min) kernels of a black colored cultivar to water extraction and HPLC analysis, a prominent peak was detected in both extracts. The structure of the substance under those peaks was identified by mass and NMR spectrometry as cyanidin 3-sambubioside in peanut testae for the first time. Subjection of cyanidin 3-sambubioside to antioxidation and anti-inflammation assessments revealed that it was a potent antioxidant and inhibitor of nitric oxide production. PMID- 19807154 TI - Endogenous levels of Echinacea alkylamides and ketones are important contributors to the inhibition of prostaglandin E2 and nitric oxide production in cultured macrophages. AB - Because of the popularity of Echinacea as a dietary supplement, researchers have been actively investigating which Echinacea constituent or groups of constituents are necessary for immune-modulating bioactivities. Our prior studies indicate that alkylamides may play an important role in the inhibition of prostaglandin E2 (PGE(2)) production. High-performance liquid chromatography fractionation, employed to elucidate interacting anti-inflammatory constituents from ethanol extracts of Echinacea purpurea, Echinacea angustifolia, Echinacea pallida, and Echinacea tennesseensis, identified fractions containing alkylamides and ketones as key anti-inflammatory contributors using lipopolysaccharide-induced PGE(2) production in RAW264.7 mouse macrophage cells. Nitric oxide (NO) production and parallel cytotoxicity screens were also employed to substantiate an anti inflammatory response. E. pallida showed significant inhibition of PGE(2) with a first round fraction, containing gas chromatography-mass spectrometry (GC-MS) peaks for Bauer ketones 20, 21, 22, 23, and 24, with 23 and 24 identified as significant contributors to this PGE(2) inhibition. Chemically synthesized Bauer ketones 21 and 23 at 1 microM each significantly inhibited both PGE(2) and NO production. Three rounds of fractionation were produced from an E. angustifolia extract. GC-MS analysis identified the presence of Bauer ketone 23 in third round fraction 3D32 and Bauer alkylamide 11 making up 96% of third round fraction 3E40. Synthetic Bauer ketone 23 inhibited PGE(2) production to 83% of control, and synthetic Bauer alkylamide 11 significantly inhibited PGE(2) and NO production at the endogenous concentrations determined to be present in their respective fraction; thus, each constituent partially explained the in vitro anti inflammatory activity of their respective fraction. From this study, two key contributors to the anti-inflammatory properties of E. angustifolia were identified as Bauer alkylamide 11 and Bauer ketone 23. PMID- 19807155 TI - Acteoside and 6-O-acetylacteoside downregulate cell adhesion molecules induced by IL-1beta through inhibition of ERK and JNK in human vascular endothelial cells. AB - Acteoside, an active phenylethanoid glycoside of many medicinal plants and bitter tea, displays anti-inflammatory properties in vitro. However, it is unclear whether acteoside and similar compounds may inhibit the expression of cell adhesion molecules (CAMs), which plays a role in the pathogenesis of atherosclerosis and inflammation. Here, we found that acteoside, isoacteoside, and 6-O-acetylacteoside inhibited IL-1beta-activated expression of intercellular CAM-1 (ICAM-1) and vascular CAM-1 (VCAM-1) in human umbilical vein endothelial cells (HUVECs); the inhibitory potency was as follows: 6-O-acetylacteoside > acteoside > isoacteoside. Acteoside and 6-O-acetylacteoside also dose-dependently inhibited VCAM-1 gene promoter activity in IL-1beta-activated HUVECs. The inhibition of acteoside and 6-O-acetylacteoside on IL-1beta-activated expression of CAMs was manifested by decreased phosphorylation of extracellular signal regulated kinase (ERK) and c-Jun N-terminal kinase (JNK). These results indicate that acteoside and 6-O-acetylacteoside may exert anti-inflammatory activities in vascular endothelium by inhibiting the expression of CAMs, primarily through decreased phosphorylation of ERK and JNK. PMID- 19807156 TI - Isolation and characterization of N-feruloyltyramine as the P-selectin expression suppressor from garlic (Allium sativum). AB - Garlic (Allium sativum) is a medicinal and culinary plant reported to have several positive health effects on cardiovascular diseases, particularly via suppressing platelet activation. Therefore, active compounds inhibiting platelet activation were isolated from garlic extract using a P-selectin expression suppressing activity-guided fractionation technique. Garlic cloves were extracted with methanol, sequentially partitioned using ethyl acetate, and n-butanol. The ethyl acetate portion was fractionated using silica gel chromatography. The fraction with highest P-selectin expression suppressing activity was further purified using HPLC, and the compounds in the fraction were analyzed using MS, MS/MS, and NMR spectroscopic methods. Using NMR spectroscopy, the compound with highest suppressing activity was confirmed as N-feruloyltyramine. At the concentration of 0.05 microM, N-feruloyltyramine was able to suppress P-selectin expression on platelets by 31% (P < 0.016). Since COX enzymes are deeply involved in the regulation of P-selectin expression on platelets, potential effects of N feruloyltyramine on COX enzymes were investigated. As expected at the concentration of 0.05 microM, N-feruloyltyramine was found to be a very potent compound able to inhibit COX-I and -II enzymes by 43% (P < 0.012) and 33% (P < 0.014), respectively. N-Feruloyltyramine is likely to inhibit COX enzymes, thereby suppressing P-selectin expression on platelets. PMID- 19807157 TI - Saponins in yerba mate tea ( Ilex paraguariensis A. St.-Hil) and quercetin synergistically inhibit iNOS and COX-2 in lipopolysaccharide-induced macrophages through NFkappaB pathways. AB - Yerba mate tea ( Ilex paraguariensis ) is growing in popularity around the world. The objective of this study was to investigate the potential anti-inflammatory effect of yerba mate tea (MT) extracts as well as some of its phytochemicals and their interactions. MT and decaffeinated MT extracts [1-300 microM chlorogenic acid (CHA) equiv]; CHA, caffeine from MT (matein), and mate saponins (1-300 microM); quercetin (1-200 microM); and ursolic and oleanolic acids (1-100 microM) were tested by measuring their ability to inhibit COX-2/PGE(2) and iNOS/NO pathways in LPS-induced RAW 264.7 macrophages. Mate saponins (IC(50) = 20 microM) and oleanolic acid (IC(50) = 80 microM) significantly inhibited iNOS/NO pathways, whereas ursolic acid showed low or no inhibition at 100 microM. Quercetin was the most potent inhibitor of pro-inflammatory responses at a concentration 10 times lower than the concentrations used of other compounds (IC(50) = 11.6 microM for NO, 7.9 microM for iNOS, and 6.5 microM for PGE(2)). Combination of quercetin/mate saponins (0.001:0.004, molar ratio) resulted in synergistic interaction inhibiting both NO and PGE(2) production. It also suppressed IL-6 and IL-1beta production and resulted in reduction of LPS-induced nuclear translocation of nuclear factor-kappaB subunits. MT extract did not have a potent anti-inflammatory effect perhaps due to the antagonistic effect of some of its compounds. However, whole MT consumption still has a promising anti-inflammatory outcome mainly through the PGE(2)/COX-2 pathway. To the authors' knowledge, this is the first study demonstrating the efficacy, interactions, and mechanisms of some MT phytochemicals in inhibiting pro-inflammatory responses. PMID- 19807158 TI - Collaborative trial validation studies of real-time PCR-based GMO screening methods for detection of the bar gene and the ctp2-cp4epsps construct. AB - Polymerase Chain Reaction (PCR)-based screening methods targeting genetic elements commonly used in genetically modified (GM) plants are important tools for the detection of GM materials in food, feed, and seed samples. To expand and harmonize the screening capability of enforcement laboratories, the German Federal Office of Consumer Protection and Food Safety conducted collaborative trials for interlaboratory validation of real-time PCR methods for detection of the phosphinothricin acetyltransferase (bar) gene from Streptomyces hygroscopicus and a construct containing the 5-enolpyruvylshikimate-3-phosphate synthase gene from Agrobacterium tumefaciens sp. strain CP4 (ctp2-cp4epsps), respectively. To assess the limit of detection, precision, and accuracy of the methods, laboratories had to analyze two sets of 18 coded genomic DNA samples of events LLRice62 and MS8 with the bar method and NK603 and GT73 with the ctp2-cp4epsps method at analyte levels of 0, 0.02, and 0.1% GM content, respectively. In addition, standard DNAs were provided to the laboratories to generate calibration curves for copy number quantification of the bar and ctp2-cp4epsps target sequences present in the test samples. The study design and the results obtained are discussed with respect to the difficult issue of developing general guidelines and concepts for the collaborative trial validation of qualitative PCR screening methods. PMID- 19807159 TI - Changes of isoflavone profile in the hypocotyls and cotyledons of soybeans during dry heating and germination. AB - A gradient reversed-phase high-performance liquid chromatography (HPLC) method has been developed to be suitable for the separation and determination of 12 isoflavones in soybeans. Profiles of daidzein, genistein, glycitein, and their malonyl-, acetyl-, and nonconjugated beta-glycosides were determined in cotyledons and hypocotyls of soybeans as affected by dry heating and germination. The results showed that the compositions and concentrations of isoflavones were remarkably different in the two parts of soybeans, and hypocotyls contained a much higher content of isoflavones than cotyledons (e.g., 7.8-fold higher). In hypocotyls, daidzein and its glycoside conjugates (59.6%) were the most abundant isoflavones, being followed by glycitein (26.6%) and genistein series (13.8%). In cotyledons, genistein and its glycoside conjugates (61.9%) were the main isoflavones, being followed by daidzein series (38.1%), and no glycitein series was found. Both hypocotyls and cotyledons contained remarkably high amounts of malonylglycosides (69.1 and 69.4%, respectively) and beta-glycosides (27.1 and 25.4%), and only a very small quantity of aglycones (3.8 and 5.2%) and no acetylglycosides were detected. Acetylglycosides and beta-glycosides were the thermal decarboxylation and deesterification products, respectively, of malonylglycosides, which were thermally unstable. The relative rates of decarboxylation and deesterification reactions were different in cotyledons and hypocotyls at different temperatures. During the process of germination, beta glycosides decreased, and malonylglycosides and aglycones increased, and then, malonylglycosides were the major fractions in germinating soybeans. Interestingly, the present study occasionally found a significant circadian change between malonylglycosides and aglycones with a nocturnal increase of aglycones and decrease of malonylglycosides during germination, and even aglycones became the most abundant forms at night. However, this mechanism is yet to be investigated. PMID- 19807160 TI - Development of a factorial design to study the effect of the major hemicellulosic sugars on the production of surface-active compounds by L. pentosus. AB - Nowadays, there are no studies about the role of the major hemicellulosic sugars on the production of surface-active compounds by Lactobacillus pentosus, although it was demonstrated that the activity of these compounds can be related to the agricultural residue from which they come, as the sugar solutions obtained from different agricultural residues contain different types and ratios of hemicellulosic sugars. Therefore, in this work, an incomplete factorial design was employed to test the relationship between the type and the ratio of hemicellulosic sugars present in hydrolysates from agricultural residues and the activity of surface-active compounds (cell-bond biosurfactants and extracellular bioemulsifiers) produced by L. pentosus. This design allowed us to establish models (that include linear, interaction, and quadratic terms) between dependent and independent variables. The independent variables used and their variation limits were as follows: glucose concentration (0-10 g L(-1)), xylose concentration (5-15 g L(-1)), and arabinose concentration (0-10 g L(-1)), whereas the 13 dependent variables studied were based on the measurement of surface tension and emulsifying capability. After the study, it was found that the emulsifier capacity of extracellular bioemulsifiers produced by L. pentosus increases at high glucose and xylose concentrations, with glucose concentration as the most influential variable in the range studied. However, the increase of glucose in the absence of xylose produced biosurfactants with low surface activity, with, in this case, the xylose concentration as the most influential variable. Taking into account the xylose/glucose ratio, the best results were obtained with xylose/glucose ratios around 1.5-3.5, which can be found in hemicellulosic hydrolysates from trimming vine shoots or grape marc hydrolysates. PMID- 19807161 TI - Characterization of the key aroma compounds in beef and pork vegetable gravies a la chef by application of the aroma extract dilution analysis. AB - By application of the aroma extract dilution analysis (AEDA) on an aroma distillate isolated from a freshly prepared, stewed beef/vegetable gravy, 52 odor active compounds were detected in the flavor dilution (FD) factor range of 4 4096. On the basis of high FD factors in combination with the results of the identification experiments, 3-(methylthio)propanal (cooked potato), 3-mercapto-2 methylpentan-1-ol (gravy-like), (E,E)-2,4-decadienal (deep-fried, fatty), 3 hydroxy-4,5-dimethyl-2(5H)-furanone (lovage-like), vanillin (vanilla-like), (E,E) 2,4-nonadienal (deep-fried), and (E)-2-undecenal (metallic) are suggested as key contributors to the aroma of the gravy. To get an insight into the role of the vegetables as sources of gravy odorants, a beef gravy was prepared without vegetables. The AEDA results revealed that, in particular, onions and leek are important sources of gravy aroma compounds, adding particularly the very potent, gravy-like smelling 3-mercapto-2-methylpentan-1-ol to the overall aroma profile. Further compounds that were clearly derived from the vegetables and, thus, are important modifiers of the overall aroma were 4-vinyl-2-methoxyphenol, (E)-beta damascenone, beta-ionone, 2-isopropyl-3-methoxypyrazine, and 2-(sec-butyl)-3 methoxypyrazine. Interestingly, none of the key odorants detected in the gravy can be assumed to be formed from a reaction between beef and vegetable constituents. A comparison of the odorants in the beef/vegetable gravy with a gravy prepared according to the same procedure, but substituting beef by pork meat, indicated that most of the aroma compounds were identical-although different in FD factors-but the tallowy smelling 12-methyltridecanal was detected as key odorant only in the beef/vegetable gravy. PMID- 19807162 TI - Effect of genotype and environment on citrus juice carotenoid content. AB - A selection of orange and mandarin varieties belonging to the same Citrus accession and cultivated in Mediterranean (Corsica), subtropical (New Caledonia), and tropical areas (principally Tahiti) were studied to assess the effect of genotype and environmental conditions on citrus juice carotenoid content. Juices from three sweet orange cultivars, that is, Pera, Sanguinelli, and Valencia ( Citrus sinensis (L.) Osbeck), and two mandarin species ( Citrus deliciosa Ten and Citrus clementina Hort. ex Tan), were analyzed by HPLC using a C(30) column. Annual carotenoid content variations in Corsican fruits were evaluated. They were found to be very limited compared to variations due to varietal influences. The statistical analysis (PCA, dissimilarity tree) results based on the different carotenoid compounds showed that citrus juice from Corsica had a higher carotenoid content than citrus juices from tropical origins. The tropical citrus juices were clearly differentiated from citrus juices from Corsica, and close correlations were obtained between beta-cryptoxanthin and phytoene (r = 0.931) and beta-carotene and phytoene (r = 0.918). More broadly, Mediterranean conditions amplified interspecific differentiation, especially by increasing the beta-cryptoxanthin and cis-violaxanthin content in oranges and beta-carotene and phytoene-phytofluene content in mandarins. Thus, at a quantitative level, environmental conditions also had a major role in determining the levels of carotenoids of nutritional interest, such as the main provitamin A carotenoids in citrus juice (beta-cryptoxanthin and beta-carotene). PMID- 19807163 TI - Microemulsions as nanoreactors to produce whey protein nanoparticles with enhanced heat stability by sequential enzymatic cross-linking and thermal pretreatments. AB - Sequential enzymatic cross-linking and heat pretreatments were used in this work to enhance the heat stability of whey protein isolate (WPI). In the first route, WPI was cross-linked by transglutaminase before incorporation in microemulsions for heat pretreatment at 90 degrees C for 20 min. In the second route, WPI was cross-linked by transglutaminase within microemulsions before thermal pretreatment. Particles produced from the two routes were different in dimension and heat stability and were also affected by the ratio of WPI and enzyme and cross-linking duration. At appropriate conditions, for example, 10 h of cross linking by transglutaminase equivalent to 5% mass of WPI using the first route, a 5% dispersion (pH 6.8 and 100 mM NaCl) of the produced nanoparticles remained clear after heating at 90 degrees C for 20 min. In comparison, nanoparticles produced by thermal pretreatment only in a microemulsion corresponded to a translucent, flowable dispersion, whereas native WPI formed a gel. This novel approach can be used to manufacture heat-stable whey protein ingredients for clear beverage applications. PMID- 19807164 TI - Kinetic analysis and mechanism on the inhibition of chlorogenic acid and its components against porcine pancreas alpha-amylase isozymes I and II. AB - Chlorogenic acid (5-caffeoylquinic acid, 5-CQA) is a kind of polyphenol and is richly included in green coffee beans. The inhibitory effects of 5-CQA and its components, caffeic acid (CA) and quinic acid (QA), on the two porcine pancreas alpha-amylase (PPA) isozymes, PPA-I and PPA-II, were investigated using p nitrophenyl-alpha-D-maltoside as substrate at pH 6.9 and 30 degrees C. The inhibition potencies of the respective inhibitors against both PPA isozymes were almost the same and in the order of 5-CQA > CA >> QA. Their IC(50) values were 0.07-0.08 mM, 0.37-0.40 mM, and 25.3-26.5 mM, respectively. The inhibition mechanisms of 5-CQA and CA were investigated by kinetic analyses, and the inhibitor constants K(i) and K(i)' (for the free enzyme and enzyme-substrate complex, respectively) were determined. It was indicated that 5-CQA and CA showed mixed-type inhibition with K(i) > K(i)' against both PPA-I and PPA-II. The binding of PPA-I or PPA-II with 5-CQA or CA was all exothermic and enthalpy driven. QA is a poor inhibitor, and its inhibitory mode was unique and hardly analyzed by a simple Michaelis-Menten-type interaction between the enzyme and inhibitor. However, it was shown that the inhibitory activity of CA was enhanced 5 times by ester-bond formation with QA in the form of 5-CQA. These results provide us with significant hints for the development of alpha-amylase inhibitors useful for the prevention of diabetes and obesity. PMID- 19807165 TI - Antifungal mechanism of a novel antifungal protein from pumpkin rinds against various fungal pathogens. AB - A novel antifungal protein (Pr-2) was identified from pumpkin rinds using water soluble extraction, ultrafiltration, cation exchange chromatography, and reverse phase high-performance liquid chromatography. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry indicated that the protein had a molecular mass of 14865.57 Da. Automated Edman degradation showed that the N-terminal sequence of Pr-2 was QGIGVGDNDGKRGKR-. The Pr-2 protein strongly inhibited in vitro growth of Botrytis cinerea, Colletotrichum coccodes, Fusarium solani, Fusarium oxysporum, and Trichoderma harzianum at 10-20 microM. The results of confocal laser scanning microscopy and SYTOX Green uptake demonstrated that its effective region was the membrane of the fungal cell surface. In addition, this protein was found to be noncytotoxic and heat-stable. Taken together, the results of this study indicate that Pr-2 is a good candidate for use as a natural antifungal agent. PMID- 19807166 TI - Dietary supplementation with alpha-amylase inhibitor wheat albumin to high-fat diet-induced insulin-resistant rats is associated with increased expression of genes related to fatty acid synthesis in adipose tissue. AB - It is well-known that insulin resistance induces lipid abnormalities by decreasing insulin actions in adipose tissue. This study examined the effects of inhibiting postprandial hyperglycemia/hyperinsulinemia, using the alpha-amylase inhibitor wheat albumin (WA), on the expression of genes related to fatty acid metabolism in the adipose tissue of high-fat diet-induced insulin-resistant rats. Postprandial glucose and insulin levels were significantly lower after oral starch loading with WA than with inactivated WA in insulin-resistant rats. In addition, the increases in the plasma triacylglycerol and insulin levels by feeding insulin-resistant rats a control diet were inhibited by WA supplementation. Supplementation with WA increased the mRNA levels of not only fatty acid synthase (FAS) and acyl-CoA carboxylase (ACC) but also their transcriptional factors such as carbohydrate response element-binding protein (ChREBP) and sterol regulatory element binding protein (SREBP)1 in the mesenteric adipose tissue of the insulin-resistant rats. In addition, supplementation with WA tended to increase the protein expression levels of FAS and ACCs. These results suggest that reductions in the plasma triacylglycerol and insulin levels by inhibiting hyperglycemia/hyperinsulinemia with the alpha-amylase inhibitor WA in high-fat diet-induced insulin-resistant rats are associated with increased expression of genes related to fatty acid synthesis and their transcriptional factors in adipose tissue. PMID- 19807167 TI - Retraction. Metabolic profiling of root exudates of Arabidopsis thaliana. PMID- 19807168 TI - A crystalline germanate with mesoporous 30-ring channels. AB - A novel germanate |C(6)N(2)H(18)|(30)[Ge(9)O(18)X(4)](6)[Ge(7)O(14)X(3)](4)[Ge(7)O(14.42)X(2.58)](8 [GeX(2)](1.73) (X = OH, F) with 30-ring channels built from Ge(7)O(14)X(3) (Ge(7)) and Ge(9)O(18)X(4) (Ge(9)) clusters was obtained under solvothermal conditions. It has a mesoporous pore of 13.0 x 21.4 A(2), which represents the largest pore ring found in crystalline open-framework materials. The framework can be described as a 3-D net structure and viewed as introducing Ge(7) and Ge(9) clusters into the 4,8-heterocoordinated csq net. PMID- 19807169 TI - Raman spectroscopy and cage occupancy of hydrogen clathrate hydrate from first principle calculations. AB - Statistically averaged Raman spectra of H(2) in hydrogen clathrate are calculated by quantum-mechanical calculations and molecular dynamics simulations. The result shows that the H(2) molecules in the large cages and singly occupied small cages are loosely encaged and the vibrational modes are softened and uncoupled, while those in the doubly occupied small cages are tightly confined, the vibrational modes are strongly coupled, and the frequencies are blue-shifted relative to the free gas. This finding provides important new insights on characterizing the cage occupancy and could inspire innovative experiments for synthesizing the clathrate as a hydrogen storage medium. PMID- 19807170 TI - Water-gas shift reaction catalyzed by redox enzymes on conducting graphite platelets. AB - The water-gas shift (WGS) reaction (CO + H(2)O <==> CO(2) + H(2)) is of major industrial significance in the production of H(2) from hydrocarbon sources. High temperatures are required, typically in excess of 200 degrees C, using d-metal catalysts on oxide supports. In our study the WGS process is separated into two half-cell electrochemical reactions (H(+) reduction and CO oxidation), catalyzed by enzymes attached to a conducting particle. The H(+) reduction reaction is catalyzed by a hydrogenase, Hyd-2, from Escherichia coli, and CO oxidation is catalyzed by a carbon monoxide dehydrogenase (CODH I) from Carboxydothermus hydrogenoformans. This results in a highly efficient heterogeneous catalyst with a turnover frequency, at 30 degrees C, of at least 2.5 s(-1) per minimum functional unit (a CODH/Hyd-2 pair) which is comparable to conventional high temperature catalysts. PMID- 19807171 TI - Electrogenerated chemiluminescence of soliton waves in conjugated polymers. AB - We report the discovery of soliton-like electrogenerated chemiluminescence (ECL) waves in the electrochemical oxidation of thin films (15-250 nm) of the semiconducting conjugated polymer poly(9,9-dioctylfluorene-co-benzothiadiazole) (F8BT). The ECL "wave fronts" were visualized and imaged in space and time by optical emission microscopy and observed to freely propagate parallel to the plane of the electrode. The data reveal that the EC solitons are triggered by the flux of electrolytes into the polymer film at nanoscale "leaks" (or defects) and propagate by a highly nonlinear, positive-feedback kinetic mechanism involving EC oxidation, mass transport of electrolyte anions, and oxidation-induced swelling, all taking place within the conjugated polymer film. Several model systems with spatially distributed reaction "triggers" were studied to elucidate the mechanistic details of this complex oxidation process in polymer films. PMID- 19807172 TI - Site preferences of fluoride guest ions in the calcium silicate phases of Portland cement from 29Si{19F} CP-REDOR NMR spectroscopy. AB - A reduction in CO(2) emission from Portland cement production can be achieved by energy savings associated with a lowering of the temperature at which the high temperature alite (Ca(3)SiO(5)) and belite (Ca(2)SiO(4)) silicates form. This can be accomplished by fluoride mineralization where a small amount of fluorine (e.g., CaF(2)) is added to the raw mix of starting materials. This work provides the mechanism for incorporation of fluoride ions in the calcium silicate phases of Portland cements which is important in the optimization of the fluoride mineralization. It is demonstrated by double-resonance (29)Si{(19)F} CP/MAS NMR experiments that the fluoride ions are exclusively incorporated into the alite phase of the two calcium silicates. The fluoride ions substitute for oxygen by a coupled mechanism that also involves replacement of Si(4+) by Al(3+) to achieve charge balance. Most importantly, (29)Si{(19)F} REDOR NMR experiments reveal that the fluoride ions are incorporated in alite with a site preference for the "interstitial" oxygen sites and thus not the covalently bonded oxygens of the SiO(4) units. This implies that only one-fifth of the oxygen sites in alite are available for substitution by fluoride ions which limits the gain in entropy of mixing that is a key factor for the reduction in upper temperature of the cement kiln. PMID- 19807173 TI - Total syntheses of isodomoic acids G and H. AB - The total syntheses of marine natural products belonging to the kainoid family, isodomoic acids G and H, are described. The strategic connection involves a sequential silylcarbocyclization/silicon-based cross-coupling process. These total syntheses were achieved efficiently via a 12- and a 13-step, longest-linear sequence, respectively. The key transformations include a diastereoselective rhodium-catalyzed carbonylative silylcarbocyclization reaction of an (l) vinylglycine-derived 1,6-enyne, a desilylative iodination reaction, as well as an alkenyl-alkenyl silicon-based cross-coupling reaction. The mechanistic insight garnered during the investigation of the iododesilylation reaction enabled stereocontrolled introduction of the iodine with either inversion or retention of double bond configuration. The invertive desilylative iodination leads to the total synthesis of isodomoic acid H, while its congener, isodomoic acid G, was obtained via a retentive iododesilylation. PMID- 19807174 TI - N-heterocyclic carbene-catalyzed hydroacylation of unactivated double bonds. AB - An intramolecular N-heterocyclic carbene (NHC)-catalyzed hydroacylation of unactivated double bonds is reported. Systematic variation of the catalyst structure revealed an N-mesitylthiazolylidene annulated with a seven-membered ring to be especially reactive. This NHC enables a unique C-C bond-forming reaction to afford substituted chroman-4-ones in moderate to excellent yields, even ones containing all-carbon quaternary centers. PMID- 19807175 TI - Development of a sila-Friedel-Crafts reaction and its application to the synthesis of dibenzosilole derivatives. AB - An intramolecular sila-Friedel-Crafts reaction was developed and applied to the synthesis of dibenzosilole derivatives. This reaction proceeds under mild conditions to afford the target in relatively high yield, indicating its availability as a versatile synthetic method. The synthesis of trisilasumanene, a silicon analogue of sumanene, was achieved using the present reaction. PMID- 19807176 TI - Crystal structures of cisplatin bound to a human copper chaperone. AB - Copper trafficking proteins, including the chaperone Atox1 and the P(1B)-type ATPase ATP7B, have been implicated in cellular resistance to the anticancer drug cisplatin. We have determined two crystal structures of cisplatin-Atox1 adducts that reveal platinum coordination by the conserved CXXC copper-binding motif. Direct interaction of cisplatin with this functionally relevant site has significant implications for understanding the molecular basis for resistance mediated by copper transport pathways. PMID- 19807177 TI - Hole-transporting periodic mesostructured organosilica. AB - Hole-transporting framework is formed by surfactant-templated sol-gel polycondensation of an electroactive phenylenevinylene-based organosilane precursor. Molecular geometry of the three-armed precursor contributes to both formation of periodic mesostructures and introduction of hole conductivity in the organosilica hybrids. Electroactive organosilicas with mesopores and large surface areas have great potentials for novel photovoltaic and photocatalytic systems. PMID- 19807178 TI - Double metallocene nanowires. AB - The structural, electronic, and magnetic properties of double metallocene nanowires PnM(2) (Pn = C(8)H(6), M = V, Cr, Mn, Co, and Ni) are investigated within the framework of gradient-corrected density functional theory. The PnMn(2) nanowire is predicted to be ferromagnetic while other nanowires are either antiferromagnitic or nonmagnetic. In addition, the PnMn(2) nanowire exhibits zigzag-like structural distortion that might result from the Peierls instability. The PnV(2) nanowire has the largest cohesive energy and shortest metal-metal distance and is likely the most stable structure among the 1D double metallocene nanowires. PMID- 19807179 TI - Postsynthetic modifications of iron-carboxylate nanoscale metal-organic frameworks for imaging and drug delivery. AB - Fe(III)-carboxylate nanoscale metal-organic frameworks (NMOFs) with the MIL-101 structure were synthesized using a solvothermal technique with microwave heating. The approximately 200 nm particles were characterized using a variety of methods, including SEM, PXRD, nitrogen adsorption measurements, TGA, and EDX. By replacing a percentage of the bridging ligand (terephthalic acid) with 2-amino terephthalic acid, amine groups were incorporated into the framework to provide sites for covalent attachment of biologically relevant cargoes while still maintaining the MIL-101 structure. In proof-of-concept experiments, an optical contrast agent (a BODIPY dye) and an ethoxysuccinato-cisplatin anticancer prodrug were successfully incorporated into the Fe(III)-carboxylate NMOFs via postsynthetic modifications of the as-synthesized particles. These cargoes are released upon the degradation of the NMOF frameworks, and the rate of cargo release was controlled by coating the NMOF particles with a silica shell. Potential utility of the new NMOF-based nanodelivery vehicles for optical imaging and anticancer therapy was demonstrated in vitro using HT-29 human colon adenocarcinoma cells. PMID- 19807180 TI - Size, shape, and internal atomic ordering of nanocrystals by atomic pair distribution functions: a comparative study of gamma-Fe2O3 nanosized spheres and tetrapods. AB - Due to their limited length of structural coherence nanocrystalline materials show very diffuse powder X-ray diffraction patterns that are difficult to interpret unambiguously. We demonstrate that a combination of high-energy X-ray powder diffraction and atomic pair distribution function analysis can be used to both assess the geometry (i.e., size and shape) and determine the internal atomic ordering of nanocrystalline materials in a straightforward way. As an example we consider cubic gamma-Fe(2)O(3) nanosized crystals shaped as spheres and tetrapods. PMID- 19807181 TI - Crystal structures of penicillin-binding protein 6 from Escherichia coli. AB - Penicillin-binding protein 6 (PBP6) is one of the two main DD-carboxypeptidases in Escherichia coli, which are implicated in maturation of bacterial cell wall and formation of cell shape. Here, we report the first X-ray crystal structures of PBP6, capturing its apo state (2.1 A), an acyl-enzyme intermediate with the antibiotic ampicillin (1.8 A), and for the first time for a PBP, a preacylation complex (a "Michaelis complex", determined at 1.8 A) with a peptidoglycan substrate fragment containing the full pentapeptide, NAM-(L-Ala-D-isoGlu-L-Lys-D Ala-D-Ala). These structures illuminate the molecular interactions essential for ligand recognition and catalysis by DD-carboxypeptidases, and suggest a coupling of conformational flexibility of active site loops to the reaction coordinate. The substrate fragment complex structure, in particular, provides templates for models of cell wall recognition by PBPs, as well as substantiating evidence for the molecular mimicry by beta-lactam antibiotics of the peptidoglycan acyl-D-Ala D-Ala moiety. PMID- 19807182 TI - Supramolecular gel based on a perylene diimide dye: multiple stimuli responsiveness, robustness, and photofunction. AB - Design of an extensive supramolecular three-dimensional network that is both robust and adaptive represents a significant challenge. The molecular system PP2b based on a perylene diimide chromophore (PDI) decorated with polyethylene glycol groups self-assembles in aqueous media into extended supramolecular fibers that form a robust three-dimensional network resulting in gelation. The self-assembled systems were characterized by cryo-TEM, cryo-SEM, and rheological measurements. The gel possesses exceptional robustness and multiple stimuli-responsiveness. Reversible charging of PP2b allows for switching between the gel state and fluid solution that is accompanied by switching on and off the material's birefringence. Temperature triggered deswelling of the gel leads to the (reversible) expulsion of a large fraction of the aqueous solvent. The dual sensibility toward chemical reduction and temperature with a distinct and interrelated response to each of these stimuli is pertinent to applications in the area of adaptive functional materials. The gel also shows strong absorption of visible light and good exciton mobility (elucidated using femtosecond transient absorption), representing an advantageous light harvesting system. PMID- 19807183 TI - Unusual electronic structure of first row transition metal complexes featuring redox-active dipyrromethane ligands. AB - Transition metal complexes (Mn --> Zn) of the dipyrromethane ligand, 1,9 dimesityl-5,5-dimethyldipyrromethane (dpm), have been prepared. Arylation of the dpm ligand alpha to the pyrrolic nitrogen donors limits the accessibility of the pyrrole pi-electrons for transition metal coordination, instead forcing eta(1),eta(1) coordination to the divalent metal series as revealed by X-ray diffraction studies. Structural and magnetic characterization (SQUID, EPR) of the bis-pyridine adducts of (dpm)Mn(II)(py)(2), (dpm)Fe(II)(py)(2), and (dpm)Co(II)(py)(2) reveal each divalent ion to be high-spin and pseudotetrahedral in the solid state, whereas the (dpm)Ni(II)(py)(2) is low-spin and adopts a square-planar geometry. Differential pulse voltammetry on the (dpm)M(II)(py)(2) series reveals a common two-electron oxidation pathway that is entirely ligand based, invariant to the divalent metal-bound, its geometry or spin state within the dpm framework. This latter observation indicates that fully populated ligand based orbitals from the dpm construct lie above partially filled metal 3d orbitals without intramolecular redox chemistry or spin-state tautomerism occurring. DFT analysis on this family of complexes corroborates this electronic structure assignment, revealing that the highest lying molecular orbitals are completely ligand-based. Chemical oxidation of the deprotonated dpm framework results in the four-electron oxidation of the dipyrrolide framework, although this oxidation product was not observed either in the electrochemical or chemical oxidation of the (dpm)M(II)(py)(2) complexes. PMID- 19807184 TI - Raman spectroelectrochemistry of molecules within individual electromagnetic hot spots. AB - The role of chemical enhancement in surface-enhanced Raman scattering (SERS) remains a contested subject. We study SERS spectra of 4-mercaptopyridine molecules excited far from the molecular resonance, which are collected from individual electromagnetic hot spots at concentrations close to the single molecule limit. The hot spots are created by depositing Tollen's silver island films on a transparent electrode incorporated within an electrochemical cell. Analysis of the intensity of the spectra relative to those obtained from individual rhodamine 6G molecules on the same surface provides a lower limit of approximately 3 orders of magnitude for the chemical enhancement. This large enhancement is likely to be due to a charge transfer resonance involving the transfer of an electron from the metal to an adsorbed molecule. Excitation at three different wavelengths, as well as variation of electrode potential from 0 to -1.2 V, lead to significant changes in the relative intensities of bands in the spectrum. It is suggested that while the bulk of the enhancement is due to an Albrecht A-term resonance Raman effect (involving the charge transfer transition), vibronic coupling provides additional enhancement which is sensitive to electrode potential. The measurement of potential-dependent SERS spectra from individual hot spots opens the way to a thorough characterization of chemical enhancement, as well to studies of redox phenomena at the single-molecule level. PMID- 19807185 TI - Size-scaling of proton conductivity in amorphous aluminosilicate acid thin films. AB - Amorphous aluminosilicate nanofilms, a-Al(0.1)Si(0.9)O(x), exhibit unique size enhancement of the proton conductivity along the thickness direction because of the presence of the zeolite-like, acid site network with the mesoscopically sized dimension inside glass matrix. The dense films with the thickness of 22-1400 nm were uniformly formed over the electrode substrate in nanometer thickness precision by multiple spin-coating with a mixed precursor sol. XANES measurements indicated that the basic framework of a-Al(0.1)Si(0.9)O(x) films was similar to the zeolitic one, consisting of the corner-linkage of SiO(4) and AlO(4) tetrahedral units. These films revealed the complex temperature- and humidity dependency of proton conductivity by the existence of two kinds of protonic carriers: Bronsted acidic protons and Lewis acidic protons. The Bronsted acidic protons could be persistent in amorphous films at around 500 degrees C, as checked by thermal desorption spectroscopy, so that the film exhibited the humidity-independent proton conductivity at temperatures above 300 degrees C. Furthermore, the conductivity across the film sigma increased in a power low by reduction of the film thickness d to less than 120 nm as sigma proportional, variant d(-tau), and it was saturated when the thickness become less than 40 nm. The observed scaling index tau was 2.2 in agreement with the value of the theoretical index (2.3) of cluster size scaling in a three-dimensional percolation system. This conduction behavior is explicable by finite size-scaling of the highly conductive pathway based on the interconnected Bronsted acid centers in the range of a few tens to hundreds of nanometers. PMID- 19807186 TI - Dynamic asymmetric multicomponent resolution: lipase-mediated amidation of a double dynamic covalent system. AB - The Strecker reaction is one of the most important multicomponent reactions developed, leading to alpha-aminonitriles that are versatile substrates for many synthetic applications. In the present study, this reaction type has been applied to a double dynamic covalent resolution protocol, leading to efficient C-C- and C N-bond generation as well as chiral discrimination. The combination of transimination with imine-cyanation enabled the dynamic exchange in more than one direction around a single stereogenic center of restricted structure. This multiple exchange process could generate a vast range of compounds from a low number of starting materials in very short time. The resulting double dynamic covalent systems, created under thermodynamic control, were subsequently coupled in a one-pot process with kinetically controlled lipase-mediated transacylation. This resulted in complete resolution of the dynamic systems, yielding the optimal N-acyl-alpha-aminonitriles for the enzyme, where the individual chemoenzymatic reactions could produce enantiomerically pure acylated N-substituted alpha aminonitriles in good yields. PMID- 19807187 TI - First-principles study of non-heme Fe(II) halogenase SyrB2 reactivity. AB - We present here a computational study of reactions at a model complex of the SyrB2 enzyme active site. SyrB2, which chlorinates L-threonine in the syringomycin biosynthetic pathway, belongs to a recently discovered class of alpha-ketoglutarate (alphaKG), non-heme Fe(II)-dependent halogenases that share many structural and chemical similarities with hydroxylases. Namely, halogenases and hydroxylases alike decarboxylate the alphaKG co-substrate, facilitating formation of a high-energy ferryl-oxo intermediate that abstracts a hydrogen from the reactant complex. The reaction mechanisms differ at this point, and mutation of active site residues (Asp for the hydroxylase to Ala or Ala to Asp/Glu for halogenase) fails to reproduce hydroxylating activity in SyrB2 or halogenating activity in similar hydroxylases. Using a density functional theory approach with a recently implemented Hubbard U correction for accurate treatment of transition metal chemistry, we explore probable reaction pathways and mechanisms via a model complex consisting of only the iron center and its direct ligands. We show that the first step, alphaKG decarboxylation, is barrierless and exothermic, but the subsequent hydrogen abstraction step has an energetic barrier consistent with that accessible under biological conditions. In the model complex we use, radical chlorination is barrierless and exothermic, whereas the analogous hydroxylation is found to have a small energetic barrier. The hydrogen abstraction and radical chlorination steps are strongly coupled: the barrier for the hydrogen abstraction step is reduced when carried out concomitantly with the exothermic chlorination step. Our work suggests that the lack of chlorination in mutant hydroxylases is most likely due to poor binding of chlorine in the active site, whereas mutant halogenases do not hydroxylate for energetic reasons. Although secondary shell residues undoubtedly modulate the overall reactivity and binding of relevant substrates, we show that a small model compound consisting exclusively of the direct ligands to the metal can help explain reactivity heretofore not yet understood in the halogenase SyrB2. PMID- 19807188 TI - Single-molecule surface-enhanced Raman spectroscopy of nonresonant molecules. AB - Single-molecule surface-enhanced Raman scattering (SERS) detection of nonresonant molecules is demonstrated experimentally using the bianalyte SERS method. To this end, bianalyte SERS is performed at 633 nm excitation using the nonresonant molecule 1,2-di-(4-pyridyl)-ethylene (BPE) in combination with a benzotriazole derivative as a partner. The results are then extended to the even more challenging case of a small nonresonant molecule, adenine, using an isotopically substituted adenine as bianalyte SERS partners. In addition, SERS cross sections of single-molecule events are quantified, thus providing estimates of the enhancement factors needed to see them. It turns out that an enhancement factor on the order of approximately 5 x 10(9) was sufficient for single-molecule detection of BPE, while maximum enhancement factors of approximately 5 x 10(10) were observed in extreme cases. In the case of adenine, single-molecule detection was only possible in the rare cases with enhancement factors of approximately 10(11). This study constitutes a quantitative fundamental test into the lowest detection limits (in terms of differential cross sections) for single-molecule SERS. PMID- 19807189 TI - Defining the mode of action of tetramic acid antibacterials derived from Pseudomonas aeruginosa quorum sensing signals. AB - In nature, bacteria rarely exist as single, isolated entities, but rather as communities comprised of many other species including higher host organisms. To survive in these competitive environments, microorganisms have developed elaborate tactics such as the formation of biofilms and the production of antimicrobial toxins. Recently, it was discovered that the gram-negative bacterium Pseudomonas aeruginosa , an opportunistic human pathogen, produces an antibiotic, 3-(1-hydroxydecylidene)-5-(2-hydroxyethyl)pyrrolidine-2,4-dione (C(12)-TA), derived from one of its quorum sensing molecules. Here, we present a comprehensive study of the expanded spectrum of C(12)-TA antibacterial activity against microbial competitors encountered by P. aeruginosa in nature as well as significant human pathogens. The mechanism of action of C(12)-TA was also elucidated, and C(12)-TA was found to dissipate both the membrane potential and the pH gradient of Gram-positive bacteria, correlating well with cell death. Notably, in stark contrast to its parent molecule 3-oxo-dodecanoyl homoserine lactone (3-oxo-C(12)-HSL), neither activation of cellular stress pathways nor cytotoxicity was observed in human cells treated with C(12)-TA. Our results suggest that the QS machinery of P. aeruginosa has evolved for a dual-function, both to signal others of the same species and also to defend against host immunity and competing bacteria. Because of the broad-spectrum antibacterial activity, established mode of action, lack of rapid resistance development, and tolerance by human cells, the C(12)-TA scaffold may also serve as a new lead compound for the development of antimicrobial therapeutics. PMID- 19807190 TI - Aza-beta3-cyclopeptides: a new way of controlling nitrogen chirality. AB - Sixteen and 24 membered aza-beta(3)-peptidic macrocycles containing a alpha hydrazinoacid or a beta(3)-aminoacid were synthesized. The conformation of these pseudopeptides was determined by using NH chemical shift analysis, NH extinction, VT-NMR experiments, and X-ray diffraction. The study shows that a stable conformation is retained between 223 and 413 K. The latter is characterized by an uninterrupted internal H-bond network and a syndiotactic arrangement of the asymmetric centers. It means that the presence of the optically pure residue acts as a conformational lock to select a single enantiomer through the cyclization by controlling the absolute configuration of all the nitrogen atoms. To our knowledge, this represents the first example of a dynamic enantioselection process involving several centers prone to pyramidal inversion. These results give a new impulsion to the control of nitrogen chirality, which remained limited to small cycles for 60 years. PMID- 19807191 TI - Sublimation of GeTe nanowires and evidence of its size effect studied by in situ TEM. AB - We report sublimation of crystalline GeTe nanowires at elevated temperatures in vacuum imaged by in situ transmission electron microscopy. The GeTe nanowires exhibit significant melting point suppression in the presence of Au contamination. A nanosized effusion cell is formed by coating the GeTe core with a SiO(2) shell, where the core can be evaporated or sublimated from the open end of the shell at high temperatures. By measuring the speed of the moving interface between the condensed and vapor phases, we determined the vaporization coefficient of these nanowires to be greater than or equal to approximately 10( 3) over a wide range of temperatures. At the final stage of the nanowire vaporization, the material loss occurs at a higher rate, which is evidence of a higher vaporization coefficient for nanosized GeTe. This in situ technique offers a quantitative method of investigating phase transition dynamics and kinetics of nanomaterials, an important topic for designing nanoscale devices to be operated at high temperatures such as phase change memory. PMID- 19807192 TI - Unveiling the chemical and morphological features of Sb-SnO2 nanocrystals by the combined use of high-resolution transmission electron microscopy and ab initio surface energy calculations. AB - Modeling of nanocrystals supported by advanced morphological and chemical characterization is a unique tool for the development of reliable nanostructured devices, which depends on the ability to synthesize and characterize materials on the atomic scale. Among the most significant challenges in nanostructural characterization is the evaluation of crystal growth mechanisms and their dependence on the shape of nanoparticles and the distribution of doping elements. This paper presents a new strategy to characterize nanocrystals, applied here to antimony-doped tin oxide (Sb-SnO(2)) (ATO) by the combined use of experimental and simulated high-resolution transmission electron microscopy (HRTEM) images and surface energy ab initio calculations. The results show that the Wulff construction can not only describe the shape of nanocrystals as a function of surface energy distribution but also retrieve quantitative information on dopant distribution by the dimensional analysis of nanoparticle shapes. In addition, a novel three-dimensional evaluation of an oriented attachment growth mechanism is provided in the proposed methodology. This procedure is a useful approach for faceted nanocrystal shape modeling and indirect quantitative evaluation of dopant spatial distribution, which are difficult to evaluate by other techniques. PMID- 19807194 TI - A model-based ensembling approach for developing QSARs. AB - Ensemble methods have become popular for QSAR modeling, but most studies have assumed balanced data, consisting of approximately equal numbers of active and inactive compounds. Cheminformatics data are often far from being balanced. We extend the application of ensemble methods to include cases of imbalance of class membership and to more adequately assess model output. Based on the extension, we propose an ensemble method called MBEnsemble that automatically determines the appropriate tuning parameters to provide reliable predictions and maximize the F measure. Results from multiple data sets demonstrate that the proposed ensemble technique works well on imbalanced data. PMID- 19807195 TI - Protein-ligand binding free energy calculation by the Smooth Reaction Path Generation (SRPG) Method. AB - We developed a new molecular dynamics simulation method for protein-ligand binding free energy calculation in an explicit water model. This method consists of three steps: (1) generation of a compound dissociation path starting from a stable protein-compound complex structure, (2) calculation of the free energy surface along the dissociation path, and (3) calculation of the free energy surface of a small area around the protein-compound complex structure, which is a free energy minimum. The protein-compound binding free energy is estimated from the information obtained by the above three steps. This method was applied to a small system, a 18-crown-6 ether with its ligand ion, and a realistic system consisting of a target protein with its inhibitor. This approximation worked well; the protein-inhibitor dissociation was successfully performed, and the binding free energies were calculated. PMID- 19807196 TI - Intestinal infections in humans in the Rocky Mountain region, United States. AB - To evaluate the seasonal prevalence of human intestinal parasites in the western states of Colorado, Utah, New Mexico, and Montana, fecal samples were examined as part of routine diagnostic testing from patients experiencing gastrointestinal discomfort in August (summer) 2006, January (winter), and April (spring) 2007. Parasite identification in positive samples was confirmed using light microscopy after wet mount and trichrome staining techniques. Seventy-eight of the 1,083 patients surveyed (7.2%) in August tested positive for at least 1 species of intestinal parasite. Forty-eight of 726 (6.6%) patients and 51 of 795 (6.4%) patients tested positive for at least 1 species in January and April, respectively. Blastocystis sp. was the most prevalent, followed by Giardia lamblia. Approximately 25% of the parasite occurrences were multiple infections involving fecal-oral transmitted species. Co-infections with Entamoeba spp. and Blastocystis sp. were common, suggesting a possible fecal-oral transmission for the latter parasite. Entamoeba spp. were more likely to co-occur than independently. Other species detected included Endolimax nana, Diphyllobothrium latum, Hymenolepis nana, Dientamoeba fragilis, and Iodamoeba butschlii. PMID- 19807197 TI - Drug discovery and development tomorrow -- changing the mindset. AB - Today's drug discovery and development paradigm is not working, and something needs to be done about it. There is good reason to believe that a move away from reliance on animal surrogates for human subjects in the Pharma Industry's R&D programmes could provide an important step forward. However, no serious move will be made in that direction until there is some hard evidence that it will be rewarded with improved productivity outcomes. The Safer Medicines Trust are proposing that a study be undertaken, involving a range of drugs that have been approved for human use, but have subsequently proved to have limitations in terms of safety and/or efficacy. The aim is to determine the efficiency of a battery of human-based test methods to identify a compound's safety and efficacy profiles, and to compare this with that of the more traditional, largely animal-based methods that were employed in their original development. Should such an approach prove more reliable, the authorities will be faced with important decisions relating to the role of human biological test data in regulatory submissions, while the Pharma Industry will be faced with the key logistical issue of how to acquire the human biomaterials necessary to make possible the routine application of such test methods. PMID- 19807198 TI - Could fresh human tissues play a key role in drug development? AB - Biopta was founded in 2002, to provide human tissue-based drug development and testing services to the pharmaceutical industry. Although animal tissues are readily available and are relatively inexpensive, they frequently fail to faithfully predict the results seen in the clinic. Human tissues can provide integrated responses to test drugs in a manner more representative than individual cell types or cell lines alone, and more-directly relevant to the species of interest -- Homo sapiens. In order to expand the use of human tissues, however, an improved infrastructure for the collection and distribution of fresh, functional tissues is highly desirable. Moreover, where there is the potential to obtain tissue from various locations, it becomes possible to test tissue that is specific to the site of drug activity. This is important, as differences may occur between the same tissue types in different locations in the body. The detection of adverse effects is greatly helped by knowledge of how existing drugs behave in the human body. These drugs can act as reference compounds, so that new compounds can then be compared, by using standard concentration-response type studies, in a huge variety of tissues, and their effects extrapolated from what is known of the reference compounds. PMID- 19807199 TI - Hurel -- an in vivo-surrogate assay platform for cell-based studies. AB - Accurate prediction of the human response to potential pharmaceuticals is difficult, often unreliable, and invariably expensive. Traditional in vitro cell culture assays are of limited value, because they do not accurately mimic the complex environment to which a drug candidate is subjected within the human body. While in vivo animal studies can account for the complex inter-cellular and inter tissue effects not observable from in vitro assays, animal studies are expensive, labour intensive, time consuming, and unpopular. In addition, there is considerable concern as to whether animal studies can predict human risk sufficiently precisely, because, first, there is no known mechanistic basis for extrapolation from high to low doses, and second, cross-species extrapolation has frequently been found to be problematic with respect to toxicity and pharmacokinetic characteristics. To address these limitations, an interactive, cell-based microfluidic biochip called a Hurel was developed. The Hurel system consists of living cells segregated into interconnected "tissue" or "organ" compartments. The organ compartments are connected by a re-circulating culture medium that acts as a "blood surrogate". The fluidics are designed so that the primary elements of the circulatory system, and more importantly, the interactions of the organ systems, are accurately mimicked. Drug candidates are exposed to a more-realistic animal or human physiological environment, thus providing a higher and more accurate informational content than can the traditional in vitro assays. By affording dynamic assessment of potential toxicity, metabolism, and bioavailability, the device's capabilities hold the potential to markedly improve the prioritisation of drug leads prior to animal studies. PMID- 19807200 TI - An immunologic model for rapid vaccine assessment -- a clinical trial in a test tube. AB - While the duration and size of human clinical trials may be difficult to reduce, there are several parameters in pre-clinical vaccine development that may be possible to further optimise. By increasing the accuracy of the models used for pre-clinical vaccine testing, it should be possible to increase the probability that any particular vaccine candidate will be successful in human trials. In addition, an improved model will allow the collection of increasingly more informative data in pre-clinical tests, thus aiding the rational design and formulation of candidates entered into clinical evaluation. An acceleration and increase in sophistication of pre-clinical vaccine development will thus require the advent of more physiologically-accurate models of the human immune system, coupled with substantial advances in the mechanistic understanding of vaccine efficacy, achieved by using this model. We believe the best viable option available is to use human cells and/or tissues in a functional in vitro model of human physiology. Not only will this more accurately model human diseases, it will also eliminate any ethical, moral and scientific issues involved with use of live humans and animals. An in vitro model, termed "MIMIC" (Modular IMmune In vitro Construct), was designed and developed to reflect the human immune system in a well-based format. The MIMIC System is a laboratory-based methodology that replicates the human immune system response. It is highly automated, and can be used to simulate a clinical trial for a diverse population, without putting human subjects at risk. The MIMIC System uses the circulating immune cells of individual donors to recapitulate each individual human immune response by maintaining the autonomy of the donor. Thus, an in vitro test system has been created that is functionally equivalent to the donor's own immune system and is designed to respond in a similar manner to the in vivo response. PMID- 19807201 TI - The use and value of in vitro technologies in metabolism studies. AB - The detailed investigation of the metabolism of drugs is one of the key issues in drug development. Several in vitro metabolism assays have been developed over the last two decades, to replace time-consuming and expensive animal studies. These have the potential to speed up drug development and increase drug safety, as they can be used to improve the prediction of the effects of drugs on humans. The key factors to be identified in metabolism are: a) the enzymes involved, and b) the metabolites produced by these enzymes. Cytochromes P450 (CYP-450s) are the key enzymes in drug metabolism. Cloning the genes encoding the CYP-450s, and the genetic engineering of suitable cells for heterologous expression, have provided new cell lines for studies on drug metabolism in vitro, under highly defined conditions. The V79 cell line, derived from Chinese hamster lung fibroblasts, was found to be suitable for heterologous expression, as these cells themselves do not express CYP-450s, thus providing a clean background for genetically engineering for the stable expression of any cloned CYP-450. In this way, V79 cell lines were created which specifically express CYP-450s from human, mouse, rat, and fish. These recombinant V79 cells have been applied in several drug metabolism and toxicity studies. PMID- 19807202 TI - Toxicogenomics in drug discovery and development -- making an impact. AB - As a branch of pharmacogenomics aimed at predicting drug safety concerns, toxicogenomics drew much excitement with the emergence of technologies such as gene expression microarrays. A few years down the line, the evidence is scant that current approaches to toxicogenomics are really making an impact in areas such as preclinical toxicology. It has been argued that there needs to be a re focus of application toward high-throughput approaches which combine the best of tissue and genomic modelling. This commentary gives a brief introduction to in vitro toxicogenomics, drawn from the perspectives of the specialist toxicogenomics company, SimuGen. PMID- 19807203 TI - Use of virtual patient populations for rescuing discontinued drug candidates and for reducing the number of patients in clinical trials. AB - The decreasing cost-efficiency of drug development threatens to result in a severe shortage of innovative drugs, which may seriously compromise patient healthcare. This risk underlines the urgency to change the paradigm in clinical research. Here, we examine a novel concept of conducting virtual clinical trials for efficiently screening drug candidates, and for evaluating their prospects of being brought to the market successfully. The virtual clinical trials are carried out by using virtual patients (denoted Optimata Virtual Patients -- OVPs). The OVP, a set of mathematical algorithms that describe the main pathological and physiological dynamic processes affected by the administered drug, has been shown to accurately predict docetaxel efficacy and safety in individual breast cancer patients. We report a test case in which virtual clinical trials have been conducted by using OVP populations for rescuing a discontinued oncology compound, ISIS-5132 (ISIS Pharmaceuticals Inc.). Our in silico study suggested that ISIS 5132 may be efficacious in combination with another drug, sunitinib malate (Sutent, Pfizer Inc.), for the treatment of prostate cancer. The recommended combined treatment is predicted to result in a higher five-year Progression-Free Survival than monotherapy with either drug alone. PMID- 19807204 TI - Monitoring tissue drug levels by clinical microdialysis. AB - The in vivo assessment of drug distribution has long been treated as a "forgotten relative" of pharmacokinetics, mainly due to a lack of appropriate methodology. Research was long restricted to the measurement of drug concentrations from biological specimens that are relatively easy to obtain, or to indirect modelling. However, data obtained by these approaches have resulted in considerable confusion about drug distribution and target site delivery, as their interpretation was flawed by several misconceptions, such as the lack of physiological input to pharmacokinetic models, the erroneous view that a tissue is a uniform matrix, and the notion that the entire drug fraction present in various tissue spaces exerts pharmacological activity. Today, drug distribution to the well defined tissue compartment -- "interstitial space fluid", the biophase for many drugs -- can be measured relatively cheaply, minimally invasively, and reproducibly, via microdialysis. PMID- 19807205 TI - The best model for humans is human -- how to accelerate early drug development safely. AB - Traditionally, the choice of which candidate compounds to take forward into development has been based on pre-clinical data. However, lack of predictivity of the human clinical situation in the models used has led to poor decision-making, and the later in the development process that such mistakes are realised, the more costly and time-consuming it is to correct them. Furthermore, compounds that may have made perfectly good drugs, have been dropped due to poor pharmacokinetics in animal models. Accelerator mass spectrometry (AMS) is an ultra-sensitive detection technique that can be used to quantify carbon-14. By administering very small amounts of (14)C-labelled compounds, AMS can be used to obtain human clinical data very early in the drug development process. Such studies: a) can be helpful in understanding human pharmacokinetics using microdosing; b) can provide early human metabolism information, to validate the choice of animal species used in pre-clinical safety testing and identify unique or disproportionate human metabolites during Phase 1; and c) can provide fundamental human pharmacokinetic data, including absolute bioavailability, by facilitating a scientifically optimal and cost-effective study design. The provision of these clinical insights at the earliest possible opportunity can lead to improved decision-making, and therefore can reduce the time and cost involved in the drug development process. PMID- 19807206 TI - De-risking drug discovery with ADDME -- avoiding drug development mistakes early. AB - The advent of early Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) screening has increased the elimination rate of weak drug candidates early in the drug-discovery process, and decreased the proportion of compounds failing in clinical trials for ADMET reasons. This paper reviews the history of ADMET screening and why it has become so important in drug discovery and development. Assays that have been developed in response to specific needs, and improvements in technology that result in higher throughput and greater accuracy of prediction of human mechanisms of toxicity, are discussed. The paper concludes with the authors' forecast of new models that will better predict human efficacy and toxicity. PMID- 19807207 TI - 21st century drug development: advances, opportunities and challenges. PMID- 19807208 TI - Can drug safety be predicted and animal experiments reduced by using isolated mitochondrial fractions? AB - Mitochondrial toxicity has resulted in the withdrawal of several drugs from the market. One particular example is nefazodone, an anti-depressant withdrawn in the USA due to hepatoxicity caused by drug-induced mitochondrial dysfunction. Drug development and safety testing can involve the use of large numbers of laboratory animals, which, without a decisive pre-screening for mitochondrial toxicity, are often unable to pre-empt higher mortality rates in some patient groups. The use of isolated mitochondria as a screening tool for drug safety can decrease the number of laboratory animals used in pre-clinical studies, thus improving animal welfare and healthcare outcomes and costs. Novel techniques involving high throughput methods can be used to investigate whether a molecule is a mitochondrial toxicant. Moreover, these screens are mechanistically-based, since the effects of the drug on oxidative phosphorylation, calcium homeostasis and mitochondrial genetics can be assessed. This review is intended to demonstrate that isolated mitochondrial fractions are suitable for predicting drug and general chemical safety in toxicological screenings, thus contributing to the refinement and reduction of animal use in laboratory research. PMID- 19807209 TI - Twenty-first century challenges for in vitro neurotoxicity. AB - During the last 40 years, studies incorporating in vitro methodologies have greatly advanced our understanding of human nerve cell biology. Attempts have been made to apply these to investigations of neurotoxicity. Due to the complexity of the nervous system, underpinned by an array of integrated interactions between a host of cell types, it is concluded that, at present, alternative neural models are most successful in determining the underlying mechanisms which can cause perturbation of normal functioning of the nervous system, both in adults and during the embryonic period. The use of tiered batteries of test models has been proposed in screening programmes for neurotoxicity, with the generation of much encouraging data in laboratories across the globe. This review aims to discuss the development of neural alternatives, considers the various model systems available, and highlights specific neuronal endpoints which can be tested, in addition to the cytotoxic evaluation of neuronal viability. Developments in molecular and stem cell biology, which are appropriate to neural tissue, and which offer the prospect of exciting advances for the next decade, are cited. PMID- 19807210 TI - The use of the Integrated Discrete Multiple Organ Co-culture (IdMOC) system for the evaluation of multiple organ toxicity. AB - The application of the Integrated Discrete Multiple Organ Co-culture (IdMOC) system in the evaluation of organ-specific toxicity is reviewed. In vitro approaches to predict in vivo toxicity have met with limited success, mainly because of the complexity of in vivo toxic responses. In vivo properties that are not well-represented in vitro include organ-specific responses, multiple organ metabolism, and multiple organ interactions. The IdMOC system has been developed to address these deficiencies. The system uses a 'wells-within-a-well' concept for the co-culturing of cells or tissue slices from different organs as physically separated (discrete) entities in the small inner wells. These inner wells are nevertheless interconnected (integrated) by overlying culture medium in the large outer containing well. The IdMOC system thereby models the in vivo situation, in which multiple organs are physically separated but interconnected by the systemic circulation, permitting multiple organ interactions. The IdMOC system, with either cells or tissue slices from multiple organs, can be used to evaluate cell type-specific or organ-specific toxicity. PMID- 19807211 TI - Immunotoxicology: opportunities for non-animal test development. AB - At present, several animal-based assays are used to assess immunotoxic effects such as immunosuppression and sensitisation. The use of whole animals, however, presents several secondary issues, including expense, ethical concerns and relevance to human risk assessment. There is a growing belief that non-animal approaches can eliminate these issues without impairing human safety, provided that biological markers are available to identify the immunotoxic potentials of new chemicals to which humans may be exposed. Driven by the 7th Amendment to the EU Cosmetics Directive, the new EU policy on chemicals (the REACH system), proposals to update the European legislation on the protection of animals used in research, and emerging visions and strategies for predicting toxicity, such in vitro methods are likely to play a major role in the near future. The realisation that the immune system can be the target of many chemicals, resulting in a range of adverse effects on the host's health, has raised serious concerns from the public and within the regulatory agencies. Hypersensitivity and immunosuppression are considered the primary focus for developing in vitro methods in immunotoxicology. However, in vitro assays to detect immunostimulation and autoimmunity are also needed. This review of the state-of-the-art in the field of in vitro immunotoxicity, reveals a lack of cell-based immunotoxicity assays for predicting the toxicity of xenobiotics toward the immune system in a simple, fast, economical and reliable way. PMID- 19807212 TI - An examination of chimpanzee use in human cancer research. AB - Advocates of chimpanzee research claim the genetic similarity of humans and chimpanzees make them an indispensable research tool to combat human diseases. Given that cancer is a leading cause of human death worldwide, one might expect that if chimpanzees were needed for, or were productive in, cancer research, then they would have been widely used. This comprehensive literature analysis reveals that chimpanzees have scarcely been used in any form of cancer research, and that chimpanzee tumours are extremely rare and biologically different from human cancers. Often, chimpanzee citations described peripheral use of chimpanzee cells and genetic material in predominantly human genomic studies. Papers describing potential new cancer therapies noted significant concerns regarding the chimpanzee model. Other studies described interventions that have not been pursued clinically. Finally, available evidence indicates that chimpanzees are not essential in the development of therapeutic monoclonal antibodies. It would therefore be unscientific to claim that chimpanzees are vital to cancer research. On the contrary, it is reasonable to conclude that cancer research would not suffer, if the use of chimpanzees for this purpose were prohibited in the US. Genetic differences between humans and chimpanzees, make them an unsuitable model for cancer, as well as other human diseases. PMID- 19807213 TI - Refinement and optimisation of the rat CFU-GM assay to incorporate the use of cryopreserved bone-marrow cells for in vitro toxicology applications. AB - The colony-forming unit-granulocyte-macrophage (CFU-GM) assay has been validated for testing drug haematotoxicity (with both mouse bone-marrow and human cord blood cells) and for predicting in vivo human Maximal Tolerated Dose (MTD) values by extrapolating in vivo data on mouse toxicity. The rat CFU-GM assay is widely used for its capability to evaluate in vitro haematotoxicity, but no standardised procedure suitable for data comparison has been developed. A validated rat CFU-GM assay is needed for many reasons - not least because the rat is the most commonly used species for the in vivo testing of toxicants. This report describes the refinement and optimisation of a standardised protocol for entering into the prevalidation phase of test development. The sensitivity of rat progenitors to granulocyte-macrophage-colony stimulating factor (GM-CSF), the correlation between the number of cells seeded and the number of colonies obtained, the role of mesenchymal cells on CFU-GM proliferation and the performance of the assay, and the effects of using different types of plastic dishes and sources of cytokines, are described. A standard operating procedure (SOP) based on the use of cryopreserved progenitors has been generated, to be applied to the in vitro toxicity testing of compounds. This SOP dramatically reduces the number of rats used and increases the homogeneity of the data obtained. PMID- 19807214 TI - The SCHER report on non-human primate research - biased and deeply flawed. AB - The European Commission's Scientific Committee on Health and Environmental Risks (SCHER) recently issued an Opinion on the need for non-human primate (NHP) use in biomedical research, and the possibilities of replacing NHP use with alternatives, as part of the Directive 86/609/EEC revision process. Here, we summarise our recent complaint to the European Ombudsman about SCHER's Opinion and the entire consultation process. It is our opinion that the Working Group almost entirely failed to address its remit, and that the Group was unbalanced and contained insufficient expertise. The Opinion presumed the validity of NHP research with inadequate supporting evidence, and ignored substantial evidence against the need for NHP research and examples of valid alternatives that could replace the use of NHPs. Because the European Commission and others might base their approach to NHP research directly on the inquiry's findings during the revision of Directive 86/609/EEC, the implications of a flawed analysis of the efficacy of NHP research are extremely serious, both for animal welfare and for human health and safety. The conduct of the SCHER inquiry, and its published Opinion, should therefore be of major and widespread concern, and should not be given any political, scientific or legislative credibility. PMID- 19807215 TI - Overcoming barriers to validation of non-animal partial replacement methods/Integrated Testing Strategies: the report of an EPAA-ECVAM workshop. AB - The use of Integrated Testing Strategies (ITS) in toxicological hazard identification and characterisation is becoming increasingly common as a method for enabling the integration of diverse types of toxicology data. At present, there are no existing procedures and guidelines for the construction and validation of ITS, so a joint EPAA WG5-ECVAM workshop was held with the following objectives: a) to investigate the role of ITS and the need for validation of ITS in the different industry sectors (pharmaceuticals, cosmetics, chemicals); b) to formulate a common definition of ITS applicable across different sectors; c) to explore how and when Three Rs methods are used within ITS; and d) to propose a validation rationale for ITS and for alternative methods that are foreseen to be used within ITS. The EPAA provided a platform for comparing experiences with ITS across different industry sectors. It became clear that every ITS has to be adapted to the product type, R&D stage, and regulatory context. However, common features of ITS were also identified, and this permitted the formulation of a general definition of ITS in a regulatory context. The definition served as a basis for discussing the needs, rationale and process of formal ITS validation. One of the main conclusions was that a formal validation should not be required, unless the strategy will serve as full replacement of an in vivo study used for regulatory purposes. Finally, several challenges and bottlenecks to the ITS validation were identified, and it was agreed that a roadmap on how to address these barriers would be established by the EPAA partners. PMID- 19807217 TI - Assessment of personal exposure to inhalable indoor and outdoor particulate matter for student residents of an academic campus (IIT-Kanpur). AB - Human exposure to particulate matter can have significant harmful effects on the respiratory and cardiovascular system. These effects vary with number, size, and chemical composition of particulate matter, which vary significantly with space and time. The Indian Institute of Technology-Kanpur (IITK), Kanpur, India, is a relatively clean academic campus in the northwest of a heavily polluted city, Kanpur. The major objectives of the study were to evaluate total exposure of fine and coarse fractions of PM(10) to a typical IITK student resident in different indoor microenvironments within the campus; to evaluate personal exposure to student residents during outdoor trips; and to evaluate personal exposure to a typical student resident carrying out routine activities. In order to account for all the sources of particulate matter exposure, measurements on several different days during the pre-monsoon season were carried out in the most common indoor microenvironments in the campus and during outdoor trips outside the campus. A 15 channel optical particle counter (model 1.108, GRIMM) was used to measure continuous real-time particle size distribution from 0.3 to 20 microm diameter. Using this instrument, exposure for 1 h at different indoor microenvironments was determined. Both the effects of location and activity, which, in turn, account for specific indoor sources and number of occupants, respectively, were carefully evaluated. Re-suspension of particles due to movement of people was found to be a major source of coarse particulate matter exposure. On the other hand, combustion sources led to elevated fine particulate levels. Chalk dust was found to be the major source of fine particulate matter in classrooms. Similar results on other sources of particulate matter are discussed in the paper. To assess the personal average size resolved particulate exposure on a student making a day trip outside the campus, study trips to most common public places in the city in a commonly preferred vehicle were made. Striking correlations between sources/activities and increase in fine and/or coarse particle concentration were clearly visible. To investigate the daily personal exposure and its relation to the activities of a typical student residing in the campus, a 24-h exposure study was done on a student who maintained a time-activity diary. The results provide insight into possible sources and their interaction with human activities in modifying the human exposure levels. A comparison between different microenvironments has been attempted for the first time in an Indian scenario using a real-time aerosol measuring instrument. PMID- 19807216 TI - Intranasal delivery of neurotrophic factors BDNF, CNTF, EPO, and NT-4 to the CNS. AB - Injury to the central nervous system (CNS) generally results in significant neuronal death and functional loss. In vitro experiments have demonstrated that neurotrophic factors such as brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), and neurotrophin-4/5 (NT-4/5) can promote neuronal survival. However, delivery to the injured CNS is difficult as these large protein molecules do not efficiently cross the blood-brain barrier. Intranasal delivery of 70 microg [(125)I]-radiolabeled BDNF, CNTF, NT-4, or erythropoietin (EPO) resulted in 0.1-1.0 nM neurotrophin concentrations within 25 min in brain parenchyma. In addition, not only did these neurotrophic factors reach the CNS, they were present in sufficient concentrations to activate the prosurvival PI3Kinase/Akt pathway, even where lower levels of neurotrophic factors were measured. Currently traumatic, ischemic and compressive injuries to the CNS have no effective treatment. There is potential clinical relevancy of this method for rescuing injured CNS tissues in order to maintain CNS function in affected patients. The intranasal delivery method has great clinical potential due to (1) simplicity of administration, (2) noninvasive drug administration, (3) relatively rapid CNS delivery, (4) ability to repeat dosing easily, (5) no requirement for drug modification, and (6) minimal systemic exposure. PMID- 19807218 TI - Antifibrotic activity of triterpenoids from the aerial parts of Euscaphis japonica on hepatic stellate cells. AB - In the course of screening antifibrotic activity of natural products employing HSC-T6, a rat hepatic stellate cell line, as an in vitro assay system, the methanolic extract of aerial parts of Euscaphis japonica (Tunb.) Kantiz (Staphyleaceae) showed significant inhibitory activity on HSC proliferation. Activity-guided fractionation led to the isolation of four triterpenoids, friedeline (1), glut-5-en-ol (2), pomolic acid (3), and methylrotundate (4). Among the triterpenoids isolated, pomolic acid (3) significantly inhibited the proliferation of HSCs at concentrations 10 and 100 microM. PMID- 19807219 TI - Insights on membrane topology and structure/function of UDP glucuronosyltransferases. AB - The main characteristic of uridine diphosphate (UDP)-glucuronosyltransferases is their potency to glucuronidate a large array of structurally unrelated substances with various nucleophilic groups. The activity of these enzymes strongly depends on their tight association to the membrane of the endoplasmic reticulum. In light of recent data, this review is focused on the membrane-assembly process, which is a prerequisite for activity, and on the amino acids that govern substrate recognition and catalysis at the active site. The major implication of the highly variable N-terminal domain of UDP-glucuronosyltransferases in the substrate specificity of these enzymes is highlighted. In the absence of crystal data of the N-terminal domain, multidisciplinary approaches of genetic-/protein engineering techniques, homology modeling with glycosyltransferases, and quantitative structure-activity relationships allowed us to point out crucial amino acids. On the basis of these results, possible reaction mechanisms for the glucuronidation of xenobiotics, involving histidine and aspartic acid residues, have been built and are discussed. PMID- 19807221 TI - The struggle to establish a research culture in the psychotherapy hospital: reflections from the Cassel Hospital experience. AB - The institutional impact and defensive reactions to the results of a longitudinal controlled outcome trial carried out within a psychotherapy hospital specializing in the treatment of personality disorder are described and discussed. The authors argue that integration of research findings with clinical practice, the development of a research culture, and building bridges with other disciplines such as psychology and neuroscience will enhance the prospect of long-term survival of psychoanalytically oriented institutions. PMID- 19807222 TI - Complicated grief and posttraumatic stress disorder in humans' response to the death of pets/animals. AB - The present exploratory project represents a cross-sectional study designed to determine the percentage of people reporting significant symptoms of complicated grief (CG) and/or posttraumatic stress disorder (PTSD) in response to the death of companion pets/animals. Human participants (N = 106) were sampled from a veterinary clinic. Fifty-two percent of participants had lost one to three pets from natural causes, 60% had never lost a pet to euthanasia, and 37% had lost one to three pets to euthanasia. The study suggests that many people experience significant attachment to their pets/animals and experience significant features of grief reactions (about 20%) after the death of a pet/animal. However, the percentage of people experiencing major pathological disruption is relatively low (<5%-12%). Thus, subclinical levels of grief and sadness are relatively common human responses to the death of companion pets/animals and last 6 months or more for about 30% of those sampled. Severe pathological reactions do occur but are quite rare among human survivors. Implications for mental health clinicians working with affected populations are discussed. PMID- 19807223 TI - An integrative approach to assessment supervision. AB - Psychological assessment supervision requires the application of technical, conceptual, and interpersonal interventions that address the supervisee's skills, knowledge, and ethical competencies. This article discusses assessment domains that are fundamental to helping trainees refine technical and reasoning skills, and utilizes Jacob, David, and Meyer's (1995) application of Johnson-Laird's (1988) typology of thought as a supervisory technique that encourages the supervisee's inductive, associative, creative, and self-reflective thinking in response to clinical situations. A clinical illustration is presented. PMID- 19807224 TI - Self-mutilation, severity of borderline psychopathology, and the Rorschach. AB - The authors explore borderline pathology on a continuum of functioning. Rorschach variables relating to (1) aggression, (2) dependency, (3) object relations, (4) defenses, and (5) boundary disturbance were measured across a nonclinical (NC) and two clinical (borderline patients without self-mutilative behavior = N-BPD, and borderline patients with self-mutilative behaviors = SM-BPD) groups. Results demonstrated good discriminate ability (87%) between clinical and nonclinical protocols. Comparisons between N-BPD and SM-BPD groups revealed overall greater pathological scores for the SM-BPD group, specifically in dependency scores. Convergence with other studies and implications for future clinical and empirical work are discussed. PMID- 19807225 TI - The reliability of the Developmental Profile based on video ratings. AB - Although psychodynamic interviews often provide a broad spectrum of interesting and useful information for the therapist, most of these interviews lack sufficient psychometric quality. So far, the Developmental Profile interview seemed an exception. However, only first-level reliability ratings based on a written account of an interview had been conducted. This study presents the results of a second-level reliability study of the Developmental Profile, based on ratings of videotapes of the Profile interview. A total of 32 videotaped interviews with psychotherapeutic inpatients were rated by three independent raters in compliance with the new version of the Developmental Profile rating manual. Contrary to earlier findings, the authors found insufficient reliability. Results are discussed. PMID- 19807229 TI - Moral distress in palliative care. PMID- 19807230 TI - Emergency department research in palliative care: challenges in recruitment. PMID- 19807231 TI - Humeral metastasis in patients with stage IV non-small-cell lung cancer portends a short life expectancy. PMID- 19807232 TI - Billing for palliative consultations a prerequisite for the field's maturation. PMID- 19807233 TI - The cautious use of cyclizine in a patient with myasthenia gravis. AB - This brief report describes the cautious but successful use of cyclizine, an anticholinergic agent, for the relief of intractable nausea in a patient with myasthenia gravis, followed by a review of the available literature. PMID- 19807234 TI - Patients who are near death are frequently unable to self-report dyspnea. AB - BACKGROUND: Standard measures of dyspnea rely on the patient's self-report. Declining consciousness and/or cognitive function and nearness to death may interfere with dyspnea reporting making the patient vulnerable to undertreatment or overtreatment. METHODS: An observational design was used with 89 consecutive patients referred for inpatient palliative care consultation. Patients were included if they were at risk for dyspnea because of one or more of the following: lung cancer, chronic obstructive pulmonary disease (COPD), heart failure, or pneumonia. Patients were asked "Are you short of breath?" and asked to quantify any distress by pointing to a visual analogue scale (VAS). Other measures included: consciousness, cognitive state, terminal illness severity, and patient demographics. RESULTS: More than half of the patients (54%) were unable to provide a yes or no response. Only 20 of 41 (49%) able to respond with yes or no were able to quantify any distress with the VAS. Ability to self-report was positively associated with consciousness (p < 0.01), cognitive state (p < 0.01), and terminal illness severity (p < 0.01). A significant inverse relationship was found between consciousness and terminal illness severity (p < 0.01). Declines in consciousness and cognitive state were strongly correlated with nearness to death (p < 0.01). CONCLUSIONS: Declining consciousness and/or cognitive state are expected when patients are near death. The ability to give even the simplest self report (yes or no) about dyspnea is lost in the near-death phase of terminal illness, yet the ability to experience distress may persist and may be overlooked and undertreated or overtreated. Other methods for symptom assessment are needed in this context. PMID- 19807235 TI - Improving the quality of spiritual care as a dimension of palliative care: the report of the Consensus Conference. AB - A Consensus Conference sponsored by the Archstone Foundation of Long Beach, California, was held February 17-18, 2009, in Pasadena, California. The Conference was based on the belief that spiritual care is a fundamental component of quality palliative care. This document and the conference recommendations it includes builds upon prior literature, the National Consensus Project Guidelines, and the National Quality Forum Preferred Practices and Conference proceedings. PMID- 19807236 TI - "South Dakota's Dying to Know": personal experiences with end-of-life care. AB - OBJECTIVE: The aim of this study was to explore end-of-life (EOL) experiences of South Dakotans who had experienced the death of a loved one in the last 5 years. METHODS: A mail questionnaire generated a return of 893 people interested in sharing their experiences about the death of a loved one in the last 5 years. From those respondents, 35 people were randomly selected for this qualitative study. Unstructured interviews were conducted, tape-recorded, and transcribed. Data were entered into the computer for analysis, and qualitative analysis was used to extract themes from the narratives. RESULTS: Participants spoke of 42 deaths, with equal thirds being positive, negative, or neutral experiences. Participants primarily discussed deaths of loved ones who were older, most typically their father or mother. Of 10 emergent themes, the most frequently cited was the importance of communication among the dying person, his or her loved ones, and health care providers. Themes revealed components of the dying experience that contributed to quality of life at end of life as well as areas for improvement. DISCUSSION: Participant reflections illuminated characteristics of a "good" death, how the health care system is understood in the face of dying, people's supportive roles in relation to the dying person, and the impact death has on survivors' own actions and preferences for EOL care. Recommendations for future research and application are discussed. PMID- 19807237 TI - Racial differences in self-reported exposure to information about hospice care. AB - BACKGROUND: Previous research suggests that lack of knowledge of hospice is a barrier to the use of hospice care by African Americans. However, there is little data examining racial differences in exposure to hospice information. OBJECTIVES: Examine racial differences in self-reported exposure to hospice information and determine how this exposure impacts beliefs about hospice care. METHODS: We surveyed 200 community-dwelling older adults (65 or older). We used Spearman's correlations to examine the relationship between responses to individual items on the Hospice Beliefs and Attitudes Scale (HBAS) and self-reported exposure to hospice information (never heard of hospice, heard a little, or heard a lot). We used multivariate analyses to examine predictors of exposure to hospice information and beliefs about hospice care (total score on HBAS). RESULTS: Compared to whites (n = 95), African Americans (n = 105) reported significantly less exposure to hospice information (p = 0.0004). Nineteen percent of African Americans and 4% of whites had never heard of hospice; 47.6% of African Americans and 71.6% of whites had heard a lot about hospice. In multivariate analysis controlling for demographics and health status, African Americans had a two times higher odds of reporting that they had never heard of hospice or heard only a little about hospice versus heard a lot about hospice (odds ratio [OR] = 2.24 [1.17, 4.27]. Greater exposure to hospice information was associated with more favorable beliefs about hospice care (outcome: total score on HBAS; parameter estimate 1.34, standard error 0.44, p = 002). CONCLUSIONS: African Americans reported less exposure to information about hospice than whites. Greater exposure to hospice information was associated with more favorable beliefs about some aspects of hospice care. Because knowledge is power, educational programs targeting older African Americans are needed to dispel myths about hospice and to provide minorities with the tools to make informed choices about end-of-life care. PMID- 19807238 TI - Medical trainee perceptions of medical school education about suffering: a pilot study. AB - BACKGROUND: The relief of suffering is a fundamental goal of medicine, but what medical students are taught about suffering has been largely unexplored. OBJECTIVE: This pilot study explored the perceptions of physicians in postgraduate training of their medical school education about suffering. DESIGN: Survey research involving physicians in postgraduate family medicine training programs. RESULTS: One hundred eighty-four of 304 surveys were returned for a response rate of 61%. Respondents perceived significant gaps in their education about the understanding and diagnosis of suffering and in their preparation to deal with the feelings engendered by caring for suffering patients. Respondents generally perceived that they were prepared to interact with suffering patients and were taught that the relief of suffering is an inherent function of being a physician, but perceived that more explicit teaching about suffering would have better prepared them for residency training. CONCLUSIONS: Perceptions of the teaching about suffering at the medical school level are quite variable with significant curricular gaps in student instruction about suffering and its relief. PMID- 19807239 TI - Systematic reviews and meta-analyses. PMID- 19807240 TI - The role of fentanyl in cancer-related pain. AB - Fentanyl is a lipophilic, short-acting, synthetic opioid with a piperidine chemical structure. Fentanyl is an effective analgesic for cancer pain, and newer formulations such as the transmucosal and buccal forms have shown efficacy for the management of cancer-related breakthrough pain. This article reviews the pharmacodynamics, pharmacology, and clinical efficacy for this new option in treating moderate to severe pain. PMID- 19807241 TI - Managing one's emotions as a clinician #203. PMID- 19807242 TI - Destination ventricular assist devices for heart failure #205. PMID- 19807244 TI - The power of presence and listening: a fellow's narrative. PMID- 19807245 TI - Three days. PMID- 19807246 TI - Treatment of malignant enterovesical fistula with octreotide. AB - Surgical treatment for internal fistula is rarely indicated for terminally ill patients with cancer because of their poor prognoses. Reports of surgical or pharmacologic treatment of vesicoenteric fistula in terminally ill patients with cancer are rare. A 73-year-old woman with rectal cancer that had directly invaded the bladder and metastasized to the liver was admitted to our hospital with high fever and severe perineal pain. Retrograde urography indicated an enterovesical fistula. Although the urinary tract infection was treatable with antibiotics, frequent episodic pain, due to urethritis secondary to the fistula, was not alleviated with opioid and topical treatment. Three days after starting octreotide 0.3 mg/d, the severe pain was alleviated, and follow-up retrograde urography revealed closure of the fistula. This suggests that treatment with octreotide may have enabled closure of the fistula. Thus, octreotide should be considered a viable therapeutic option in terminally ill patients with inoperable internal fistula. PMID- 19807254 TI - How much bone for the buck? The importance of compliance issues in economic evaluations of bisphosphonates. PMID- 19807253 TI - The effects of ERK1/2 inhibitor on the chondrogenesis of bone marrow- and adipose tissue-derived multipotent mesenchymal stromal cells. AB - This study tested the hypothesis that mitogen-activated protein kinase inhibitors suppress hypertrophy and enhance chondrogenesis during chondrogenesis of multipotent mesenchymal stromal cells (MSCs). The effects of PD98059 (an extracellular signal-regulated kinase-1/2 inhibitor) and SB203580 (a p38 inhibitor) were tested on bone marrow-derived MSCs (BMMSCs) and adipose tissue derived MSCs (ATMSCs). In vitro pellet cultures were carried out using 2.5 x 10(5) MSCs in a chondrogenic medium containing 5 ng/mL of transforming growth factor-beta(2) (TGF-beta(2)) for BMMSCs, and 5 ng/mL of TGF-beta(2) and 100 ng/mL of bone morphogenetic protein-7 (BMP-7) for ATMSCs. From the 14th day of culture, the pellets were additionally treated with PD98059 or SB203580. After 14 more days of in vitro culture, pellets were harvested for analysis. PD98059 increased DNA content and glycosaminoglycan amount in BMMSCs and ATMSCs, whereas SB203580 had little effect. Collagen type I (COL1A1) mRNA decreased to almost a quarter in BMMSCs treated with PD98059. The mRNA levels of collagen type II (COL2A1) and SRY (sex determining region Y)-box 9 (SOX-9) increased several fold in both cells after PD98059 treatment, whereas SB203580 had only a slight effect. The gene expression of collagen type X (COL10A1) and runt-related transcription factor 2 (Runx-2) decreased by half after PD98059 treatment in BMMSCs, and decreased further in ATMSCs. SB203580 elevated COL10A1 and Runx-2 gene expression in both cell types. Safranin-O staining and immunohistochemistry generally mirrored findings from real-time PCR except for diminished expression of type I collagen in ATMSCS, and more pronounced decrease in type X collagen and Runx-2 in BMMSCs after PD98059 treatment. Our study demonstrated that PD98059 suppressed hypertrophy and promoted chondrogenesis of MSCs, and provides a ground for using them in cartilage tissue engineering. PMID- 19807256 TI - Reimbursement for high-priced innovative medical devices in a DRG-based hospital care system: the German situation. AB - The implementation of high-priced innovative medical technologies in a diagnosis related group-type hospital-care reimbursement system is a challenge. If the extra costs are not reimbursed, there will be no financial incentive for a hospital to use a new technology, except for marketing reasons and research projects. A continuous in-time adaptation of the catalog of reimbursed items is necessary. PMID- 19807257 TI - Cost-effectiveness of epoetin alfa and darbepoetin alfa in patients with chemotherapy-related anemia. AB - Anemia frequently develops during cancer chemotherapy and contributes to a variety of adverse clinical outcomes. Correction of anemia has been shown to improve patient outcomes; however, the cost considerations of supportive-care interventions have also become increasingly important to payors. Chemotherapy related anemia can be managed via red blood cell transfusion; however, inherent transfusion-associated risks (e.g., infectious transmission and immunosuppression) can be particularly problematic in patients with cancer. Epoetin alfa and darbepoetin alfa, alternatives to blood transfusion in patients with chemotherapy-related anemia, are both effective at improving hemoglobin levels and transfusion requirements in patients actively receiving chemotherapy. Determining the relative cost-effectiveness of these erythropoietic therapies is an active research area. PMID- 19807258 TI - Quality of life and costs for different treatment strategies for rheumatoid arthritis. AB - Rheumatoid arthritis is a chronic, inflammatory, systemic disease. In the past, treatment (strategy) for this disease has changed dramatically, becoming more aggressive and new drugs have become available. Furthermore, closely monitoring the disease and treatment has been advocated. Rheumatoid arthritis has an extensive impact on quality of life and the cost of the disease to society is high. Since rheumatoid arthritis is a chronic disease with life-long treatment, the long-term assessment of cost-effectiveness of new treatment (strategies) frequently implies modeling. This article reviews the assessment of the diagnosis, disease process and outcome (including quality of life and costs) and methodology of cost-effectiveness (modeling) studies in rheumatoid arthritis. Furthermore, it describes the recent trends in the treatment of rheumatoid arthritis and summarizes current evidence regarding the effects on quality of life, costs and cost-effectiveness of these new treatment strategies. Since traditional disease-modifying antirheumatic drug treatment is inexpensive, early aggressive treatment with these drugs is probably cost effective since these strategies do not appear to result in elevated toxicity and are usually found to be effective. Also, closely monitoring patients, if successful, is probably cost effective. The exact place (i.e., as a first-, second- or third-line drug) of new drugs for the treatment of rheumatoid arthritis remains somewhat controversial, since cost-effectiveness analyses have varying results and methodology. Expected future developments in the field are also discussed. PMID- 19807259 TI - Adherence, asthma control, generic and disease-specific quality-of-life instruments in asthma. AB - Prior studies have investigated the association between generic health-related quality of life and medication adherence. The objective of this study was to assess the association between medication adherence and health-related quality of life using both a disease-specific and generic metric in a community setting. Overall, the authors' study findings show no association between adherence and health-related quality of life, supporting the results by Cote and colleagues that factors other than medication adherence are important in explaining health related quality of life. Furthermore, adherence was not significantly associated with asthma severity, although the strong association between asthma control and health-related quality of life, and asthma control and adherence indicates that asthma control is an important vital sign. PMID- 19807260 TI - Health status and costs of exacerbations of chronic bronchitis and COPD: how to improve antibiotic treatment. AB - Up to 10% of the adult population in most countries may be affected by chronic bronchitis or chronic obstructive pulmonary disease. The course of the disease is characterized by frequent exacerbations with increased respiratory symptoms. Exacerbations are the most important cause of morbidity and mortality, and significantly impair the health status of chronic bronchitis and chronic obstructive pulmonary disease patients. It has been observed that impairment in health status associated with exacerbations may not recover completely if recurrent exacerbations occur. Therefore, strategies to prolong the time free of exacerbations are a cornerstone of therapy. Antibiotics have demonstrated additional benefits to bronchodilators and corticosteroids in the treatment of exacerbations. Furthermore, the possible role of antibiotics in preventing exacerbations and reducing the costs of chronic bronchitis and chronic obstructive pulmonary disease patients is discussed. PMID- 19807261 TI - Antiallergic drugs and quality of life. AB - Allergic diseases represent a global health problem with a substantial impact on medical costs and socioeconomic outcomes. Furthermore, as well documented in the recent literature, these pathologies also have negative consequences on the functional, emotional and psychosocial aspects of daily life. Using validated and standardized tools, it is possible to evaluate individual outcomes. At present, the health-related quality of life parameter is used in clinical trials, population studies and public health, providing a comprehensive picture of both the disease and its treatment on the quality of life. While many studies have addressed this topic in respiratory allergy, further studies are needed to explore the use of health-related quality of life both before and after treatment in other allergic conditions such as dermatitis, eczema, urticaria and food allergy. PMID- 19807262 TI - Outcomes associated with pharmacotherapy in overactive bladder. AB - Overactive bladder is one of the most prevalent medical conditions in the USA. Prevalence rates of overactive bladder are higher in women and increase with advanced age. Although the disease is not associated with mortality, it has significant impact on health-related quality of life amongst patients. Pharmacologic, as well as behavioral treatment therapies, are available for overactive bladder treatment with pharmacologic agents being the mainstay therapy. The purpose of this review is to outline the recent developments in the treatment of overactive bladder. This is an extensive review that provides important information on the outcomes associated with different pharmacotherapeutic agents used in overactive bladder. It concludes with the authors' opinions and suggestions for the future research in this area. PMID- 19807263 TI - Economic burden of uncomplicated urinary tract infections: direct, indirect and intangible costs. AB - A better understanding of how antimicrobial therapies affect the total cost of lower uncomplicated urinary tract infection, including direct (e.g., doctor visits), indirect (e.g., productivity) and intangible (e.g., pain) costs would facilitate selection of an optimal therapeutic approach. The results of this literature review indicate that the there is a considerable societal burden from uncomplicated urinary tract infection, with approximately USD 1 billion in indirect and over USD 600 million in direct costs in 1995. However, no single identified study incorporated all three cost components, there are gaps in the knowledge concerning the current extent of these costs, and there are no comparative assessments based on total cost. Research is needed to provide current insights on the burden of uncomplicated urinary tract infection in terms of direct, indirect and intangible costs. PMID- 19807264 TI - Use of needle-free injection systems to alleviate needle phobia and pain at injection. AB - Needle phobia affects at least 10% of the general population. Subcutaneous injections are used for many reasons, including immunizations, administration of medications such as insulin and heparin, and to provide local anesthesia, both for surgery and for intravenous cannulation. Whatever the reason for its application, the injection itself may cause discomfort and/or pain. In children, in patients with needle phobia, in those who require frequent intravenous cannulations, or in those who need daily medication, the pain at injection can reach unbearable intensity that could lead to refusal of medical care. Various approaches are employed to alleviate the pain caused by intravenous cannulation. These include the use of topical analgesia [i.e., EMLA, Ametop (tetracaine], Numby Stuff and ethylchloridespray], skin infiltration with lidocaine using 25-30 gauge needles and jet injectors. This article will review the complex topic of needle phobia and needle pain, and will summarize the currently available alternatives and the new developments intended to reduce the intensity of injection pain. PMID- 19807265 TI - Applicability of discrete-choice methods to economic evaluations of complementary and alternative medicine. AB - This review discusses the potential contributions of discrete-choice methods of stated preference assessment to future economic evaluations of complementary and alternative medicine. The manner in which some complementary and alternative medicine practitioners and researchers have framed the concept of therapeutic benefits including process-related benefits is first discussed. The mechanics of a discrete-choice exercise including assessment of health state utilities and willingness to pay are then described. Potential limitations of the discrete choice method are discussed. This review concludes with a discussion of future research opportunities. The author suggests that discrete-choice methods may be integrated into economic evaluations through the benefit assessment process, particularly within the complementary and alternative medicine context. PMID- 19807266 TI - Using health-related quality of life to predict and manage pediatric healthcare. AB - Increasing healthcare costs and the prevalence of managed care make population health management an imperative. Health-related quality of life (HRQOL) is a multidimensional construct that includes both physical and psychosocial (i.e., social, emotional and role) dimensions. Early studies suggest that HRQOL can predict costs of care for pediatric populations. A key issue is how to manage the care of those identified as high need. Here again, HRQOL measurement can be useful. HRQOL measurement in the clinical setting can streamline and structure the clinical interview, potentially leading to enhanced assessment. It can also make it easier for busy pediatricians to explore and address issues of psychosocial functioning. A particularly promising area for HRQOL is in identifying, proactively, suitable candidates for case management in large enrolled populations. Further research should extend the initial studies on HRQOL predicting utilization and cost, more thoroughly specify the proportion of identified costs that are manageable and care management's effect on care for different groups of children, document the causal links between physiologic variables and HRQOL on one hand and patient functioning on the other, and understand the conditions necessary for HRQOL assessment to affect clinical practice. PMID- 19807267 TI - Prospect theory in the valuation of health. AB - Prospect theory is the prominent nonexpected utility theory in the estimation of health state preference scores for quality-adjusted life year calculation. Until recently, the theory was not considered to be developed to the point of implementation in economic analysis. This review focuses on the research and evidence that tests the implementation of prospect theory into health state valuation. The typical application of expected utility theory assumes that a decision maker has stable preferences under conditions of risk and uncertainty. Under prospect theory, preferences are dependent on whether the decision maker regards the outcome of a choice as a gain or loss, relative to a reference point. The conceptual preference for standard gamble utilities in the valuation of health states has led to the development of elicitation techniques. Empirical evidence using these techniques indicates that when individual preferences are elicited, a prospect theory consistent framework appears to be necessary for adequate representation of individual health utilities. The relevance of prospect theory to policy making and resource allocation remains to be established. Societal preferences may not need the same attitudes towards risks as individual preferences, and may remain largely risk neutral. PMID- 19807268 TI - A novel bioreactor for stimulating skeletal muscle in vitro. AB - For over 300 years, scientists have understood that stimulation, in the form of an electrical impulse, is required for normal muscle function. More recently, the role of specific parameters of the electrical impulse (i.e., the pulse amplitude, pulse width, and work-to-rest ratio) has become better appreciated. However, most existing bioreactor systems do not permit sufficient control over these parameters. Therefore, the aim of the current study was to engineer an inexpensive muscle electrical stimulation bioreactor to apply physiologically relevant electrical stimulation patterns to tissue-engineered muscles and monolayers in culture. A low-powered microcontroller and a DC-DC converter were used to power a pulse circuit that converted a 4.5 V input to outputs of up to 50 V, with pulse widths from 0.05 to 4 ms, and frequencies up to 100 Hz (with certain operational limitations). When two-dimensional cultures were stimulated at high frequencies (100 Hz), this resulted in an increase in the rate of protein synthesis (at 12 h, control [CTL] = 5.0 + or - 0.16; 10 Hz = 5.0 + or - 0.07; and 100 Hz = 5.5 + or - 0.13 fmol/min/mg) showing that this was an anabolic signal. When three-dimensional engineered muscles were stimulated at 0.1 ms and one or two times rheobase, stimulation improved force production (CTL = 0.07 + or - 0.009; 1.25 V/mm = 0.10 + or - 0.011; 2.5 V/mm = 0.14146 + or - 0.012; and 5 V/mm = 0.03756 + or - 0.008 kN/mm(2)) and excitability (CTL = 0.53 + or - 0.022; 1.25 V/mm = 0.44 + or - 0.025; 2.5 V/mm = 0.41 + or - 0.012; and 5 V/mm = 0.60 + or - 0.021 V/mm), suggesting enhanced maturation. Together, these data show that the physiology and function of muscles can be improved in vitro using a bioreactor that allows the control of pulse amplitude, pulse width, pulse frequency, and work-to-rest ratio. PMID- 19807269 TI - The stress response of the highly social African cichlid Neolamprologus pulcher. AB - In group-living species, dominant individuals are frequently aggressive toward subordinates, and such dominant aggression can lead to chronic stress, higher glucocorticoid levels, and decreased fitness for subordinates. However, in many cooperatively breeding species, it is surprisingly the dominants rather than the subordinates that exhibit higher levels of glucocorticoids, a possible consequence of the demands of maintaining high social rank and socially suppressing the reproduction of other group members. This study investigates the relationship between social status and circulating plasma cortisol in groups of the cooperatively breeding African cichlid Neolamprologus pulcher. Baseline (resting) levels of cortisol were quantified, as was the cortisol response following an acute stressor. Dominants had the higher cortisol concentrations, and these were not related to their social behavior. Cortisol concentrations correlated (positively) with social behaviors and general activity levels only in subordinate males, arguably the individuals with the least stability in the social group. No status-dependent differential responses to acute stress were detected, suggesting that the status-induced chronic stress has little effect on the capacity to mount a full stress response to large-scale, life-threatening risk. PMID- 19807270 TI - A distinct microRNA signature for definitive endoderm derived from human embryonic stem cells. AB - Human embryonic stem cells (hESCs) have the potential to differentiate into many adult cell types, and they are being explored as a resource for cell replacement therapies for multiple diseases. In order to optimize in vitro differentiation protocols, it will be necessary to elucidate regulatory mechanisms that contribute to lineage specification. MicroRNAs (miRNAs) are emerging as key regulators of hESC differentiation and embryonic development. In this study, we compare miRNA expression profiles between pluripotent hESCs and definitive endoderm (DE), an early step in the pathway toward the pancreatic lineage. Results from microarray analysis showed that DE can be distinguished by its unique miRNA profile, which consists of 37 significantly down-regulated and 17 up regulated miRNAs in 2 different cell lines and in the presence/absence of feeder layers. Comparison to other hESC-derived lineages showed that most of the highly up-regulated miRNAs are specific to endoderm in early development. Notably, miR 375, which was previously implicated in regulating development and function of later stages of pancreatic development, is highly and specifically up-regulated during DE formation, suggesting that it may have a distinct role very early in development. Examination of potential mRNA targets showed that TIMM8A is repressed by ectopic miR-375 expression in pluripotent hESCs. PMID- 19807271 TI - Strongyloidiasis in transplant patients. AB - Strongyloides stercoralis is an intestinal nematode that can persist in the human host for decades after the initial infection and can progress to fulminant hyperinfection syndrome in immunocompromised hosts. We describe a patient who died of Strongyloides hyperinfection syndrome 2 months after orthotopic heart transplantation and discuss approaches to prevention, diagnosis, and treatment. Current practice guidelines recommend screening for and treatment of Strongyloides infection before transplantation, but physicians in the United States often miss opportunities to identify patients with chronic strongyloidiasis. Screening tests have limitations, and clinical suspicion remains an important component of the evaluation before transplantation. After immunocompromised patients develop hyperinfection syndrome, diagnosis is often delayed and mortality is high, so emphasis must be placed on screening and treatment before transplantation. We review current strategies for prevention, diagnosis, and treatment of chronic intestinal strongyloidiasis in patients who will undergo transplantation and discuss the clinical features and management of Strongyloides hyperinfection syndrome in transplant recipients. PMID- 19807273 TI - The clinical spectrum of herpes simplex viremia. PMID- 19807272 TI - Clinical correlates of herpes simplex virus viremia among hospitalized adults. AB - BACKGROUND: Quantification of herpes simplex virus (HSV) DNA in the peripheral blood is often used to evaluate patients suspected of having disseminated HSV infection. Few studies have examined the clinical correlates of HSV viremia among adults. METHODS: We conducted a retrospective analysis of blood samples sent to a molecular virology reference laboratory at a university hospital for quantification of HSV DNA from October 2001 through June 2006. Medical records of patients with detectable HSV DNA were reviewed to abstract relevant clinical characteristics. RESULTS: HSV DNA was detected in 38 (4%) of 951 samples from 29 patients, 19 of whom (66%) were >16 years old. Detailed medical records were available for review from 13 (68%) of 19 adult patients. Of the 10 patients whose HSV infection was typed, 6 (60%) had HSV-2, 3 (30%) had HSV-1, and 1 (10%) had evidence of HSV-1 and HSV-2 coinfection. All patients with viremia were treated with antiviral medications. The most common clinical findings were hepatitis (62%), fever (54%), central nervous system alterations (46%), skin lesions (38%), abdominal pain (31%), and sepsis (31%). Respiratory failure (23%) was uncommon. Patients with HSV viremia were observed to have a high mortality rate (6 of 10 immunocompromised and 1 of 3 immunocompetent individuals). CONCLUSIONS: HSV viremia may be associated with a variety of signs and symptoms of morbidity in immunocompetent and immunocompromised hospitalized adults and is associated with high rates of mortality, although causality can be determined only by additional studies. PMID- 19807274 TI - Dissecting the group of common variable immunodeficiency disorders. PMID- 19807275 TI - Retention in opioid substitution treatment: a major predictor of long-term virological success for HIV-infected injection drug users receiving antiretroviral treatment. AB - BACKGROUND: The positive impact of opioid substitution treatment (OST) on opioid dependent individuals with human immunodeficiency virus (HIV) infection is well documented, especially with regard to adherence to highly active antiretroviral therapy (HAART). We used the data from a 5-year longitudinal study of the MANIF 2000 cohort of individuals infected with HIV (as a result of injection drug use) and receiving HAART to investigate the predictors of long-term virological success. Design. Data were collected every 6 months from outpatient hospital services delivering HIV care in France. We selected all patients who were receiving HAART for at least 6 months (baseline visit) and who had indications for OST (ie, still dependent on opioids). We selected a total of 113 patients, accounting for a total of 562 visits for all the analyses. METHODS: Long-term virological success was defined as an undetectable viral load after at least 6 months on HAART. Retention in OST was defined as the time interval between the last initiation or reinitiation of OST during HAART follow-up and any given visit on OST. A mixed logistic model was used to identify predictors of long-term virological success. RESULTS: At baseline, 53 patients were receiving buprenorphine, 28 patients were receiving methadone, and 32 patients were not on OST. The median duration of OST was 25 months (range, 3-42 months). In the multivariate analysis, after adjustment for significant predictors of long-term virological success such as adherence to HAART and early virological response, retention in OST was associated with long-term virological success (odds ratio, 1.20 per 6-month increase; 95% confidence interval, 1.09-1.32). CONCLUSIONS: Our study presents important evidence of the positive impact of retention in OST on HIV outcomes. Increasing access to OST based on a comprehensive model of care for HIV-infected patients who have indications for OST may foster adherence and ensure long-term response to HAART. PMID- 19807276 TI - Pharmacokinetic evaluation of rifabutin in combination with lopinavir-ritonavir in patients with HIV infection and active tuberculosis. AB - BACKGROUND: Human immunodeficiency virus (HIV)-associated tuberculosis is difficult to treat, given the propensity for drug interactions between the rifamycins and the antiretroviral drugs. We examined the pharmacokinetics of rifabutin before and after the addition of lopinavir-ritonavir. METHODS: We analyzed 10 patients with HIV infection and active tuberculosis in a state tuberculosis hospital. Plasma was collected for measurement of rifabutin, the microbiologically active 25-desacetyl-rifabutin, and lopinavir by validated high performance liquid chromatography assays. Samples were collected 2-4 weeks after starting rifabutin at 300 mg thrice weekly without lopinavir-ritonavir, 2 weeks after the addition of lopinavir-ritonavir at 400 and 100 mg, respectively, twice daily to rifabutin at 150 mg thrice weekly, and (if rifabutin plasma concentrations were below the normal range) 2 weeks after an increase in rifabutin to 300 mg thrice weekly with lopinavir-ritonavir. Noncompartmental and population pharmacokinetic analyses (2-compartment open model) were performed. RESULTS: Rifabutin at 300 mg without lopinavir-ritonavir produced a low maximum plasma concentration (C(max)) in 5 of 10 patients. After the addition of lopinavir-ritonavir to rifabutin at 150 mg, 9 of 10 had low C(max) values. Eight patients had dose increases to 300 mg of rifabutin with lopinavir-ritonavir. Most free rifabutin (unbound to plasma protein) C(max) values were below the tuberculosis minimal inhibitory concentration. For most patients, values for the area under the plasma concentration-time curve were as low or lower than those associated with treatment failure or relapse and with acquired rifamycin resistance in Tuberculosis Trials Consortium/US Public Health Service Study 23. One of the 10 patients experienced relapse with acquired rifamycin resistance. CONCLUSION: The recommended rifabutin doses for use with lopinavir-ritonavir may be inadequate in many patients. Monitoring of plasma concentrations is recommended. PMID- 19807277 TI - Late-onset combined immune deficiency: a subset of common variable immunodeficiency with severe T cell defect. AB - BACKGROUND: Common variable immunodeficiency (CVID) is a primary immune deficiency defined by defective antibody production. In most series, a small proportion of patients present with opportunistic infections (OIs). METHODS: The French DEFI study has enrolled patients with primary hypogammaglobulinemia and allows a detailed clinical and immunologic description of patients with previous OIs and/or at risk for OIs. RESULTS: Among 313 patients with CVID, 28 patients (8.9%) presented with late-onset combined immune deficiency (LOCID), defined by the occurrence of an OI and/or a CD4(+) T cell count <200 x 10(6) cells/L, and were compared with the remaining 285 patients with CVID. The patients with LOCID more frequently belonged to consanguineous families (29% vs 8%; P = .004). They differed from patients with CVID with a higher prevalence of splenomegaly (64% vs 31%), granuloma (43% vs 10%), gastrointestinal disease (75% vs 42%), and lymphoma (29% vs 4%). Even on immunoglobulin substitution, they required more frequent antibiotics administration and hospitalization. Lymphocyte counts were lower, with a marked decrease in CD4(+) T cell counts (158 x 10(6) vs 604 x 10(6) cells/L; P < .001) and a severe defect in naive CD45RA(+)CCR7(+)CD4(+) T cell counts (<20% of total CD4(+) T cells in 71% of patients with LOCID vs 37% of patients with CVID; P = .001). The CD19(+) B cell compartment was also significantly decreased (20 x 10(6) vs 102 x 10(6) cells/L; P < .001). CONCLUSIONS: LOCID differs from classic CVID in its clinical and immunologic characteristics. Systematic T cell phenotype may help to discriminate such patients from those with CVID. Identification of this phenotype should result in a more fitted diagnostic and therapeutic approach of infections and could provide insights for genetic diagnosis. PMID- 19807278 TI - Disparity in the in vitro versus in vivo regulation of fibronectin-binding proteins by 2 global regulators, saeRS and sigB, in Staphylococcus aureus. AB - Fibronectin-binding protein A plays an important role in Staphylococcus aureus endovascular infections. We characterized the sigB-saeRS-fnbA expression network with fibronectin binding in vitro and in an experimental infective endocarditis (IE) model using parental strains RN6390 and SH1000 and their respective isogenic saeRS mutants. In contrast to the in vitro data, there was no influence of saeRS on fnbA expression in the IE model, yet ex vivo fibronectin binding was reduced in saeRS mutants. Moreover, as opposed to the in vitro findings, sigB appeared to have a positive rather than a negative effect on saeRS expression within cardiac vegetations. PMID- 19807279 TI - Plasmodium falciparum multidrug resistance protein 1 and artemisinin-based combination therapy in Africa. AB - Plasmodium falciparum response mechanisms to the major artemisinin-based combination therapies (ACTs) are largely unknown. Multidrug-resistance protein (MRP)-like adenosine triphosphate (ATP)-binding cassette transporters are known to be related to multidrug resistance in many organisms. Therefore, we hypothesized that sequence variation in pfmrp1 can contribute to decreased parasite sensitivity to ACT. Through sequencing of the pfmrp1 open reading frame for 103 geographically diverse P. falciparum infections, we identified 27 single nucleotide polymorphisms (SNPs), of which 21 were nonsynonymous and 6 synonymous. Analyses of clinical efficacy trials with artesunate-amodiaquine and artemether lumefantrine detected a specific selection of the globally prevalent I876V SNP in recurrent infections after artemether-lumefantrine treatment. Additional in silico studies suggested an influence of variation in amino acid 876 on the ATP hydrolysis cycle of pfMRP1 with potential impact on protein functionality. Our data suggest for the first time, to our knowledge, the involvement of pfMRP1 in P. falciparum in vivo response to ACT. PMID- 19807282 TI - Non-invasive determination of instantaneous brachial blood flow using the oscillometric method. AB - The oscillometric method has been widely used to measure arterial systolic and diastolic blood pressures, but its potential for arterial blood flow measurements still remains to be explored. The aim of this study was to non-invasively determine arterial blood flow using an oscillometric blood flow measurement system. The system consists of a pneumatic elastic cuff, an air-pumping motor, a releaser valve, a pressure transducer, and an airflow meter. To build a non linear cuff model, we measured airflow pumped into the pneumatic cuff and cuff pressure using an airflow meter and pressure transducer during the inflation period, respectively. During the deflation period, only the pressure transducer was used to record cuff pressure. Based on the cuff model, the oscillometric blood flow waveform was obtained by integrating the oscillometric pressure waveform. We compared arterial blood flow derived from the maximum amplitude of the oscillometric blood flow waveform with Doppler-measured blood flow calculated with the diameters and blood velocities of the brachial arteries in 32 subjects who underwent diagnostic evaluations for peripheral arterial embolism. A linear correlation coefficient of r = 0.716 was found between the oscillometry- and Doppler-based blood flow measurements in the 32 subjects. These results suggest that blood flow passing through the brachial artery can be quantified non invasively using the oscillometric approach after appropriate calibration. PMID- 19807283 TI - Direct measurement of myocardial oxygen tension and high energy phosphate content under varying ventilatory conditions in rabbits. AB - Effective myocardial oxygen supply should not be compromised during cardiac surgery as it is essential to avoid circulatory and cardiac dysfunction. Local measurement of myocardial oxygen partial pressure (pO2) was therefore introduced into the operative monitoring of myocardial ischemia. The aim of the present study was to assess whether myocardial oxygen partial pressure correlates with the content of high energy phosphates (HEPs). Seven male rabbits were examined in parallel with measurement of myocardial pO2 by an implanted Clark electrode and 31phosphorus-NMR spectroscopy. The ventilatory management established hyperoxygenation followed by systemic hypoxia with hypercapnia for 20 min. Additionally, analysis of end-expiratory gas composition in combination with blood gas analysis was performed simultaneously, and hemodynamic parameter was recorded. Under hypoxic conditions the cardiovascular system was severely compromised, whereas the myocardial pO2 was only moderately impaired (pO2M 45.0+/ 16.0 mm Hg). Immediately before cardiac arrest, low values of arterial and venous pO2 were found (17.6+/-6.0 and 12.9+/-6.1 mm Hg). In contrast to near normal myocardial pO2, HEP content in the myocardium was considerably reduced and inorganic phosphorus was increased. Artificial ventilation leading to systemic hypoxia and eventually circulatory arrest resulted in almost normal myocardial pO2 but severely compromised HEP content. This somewhat unexpected finding requires further clarification, but is in accordance with findings reported previously where regulatory mechanisms have been shown to play a role in the pathophysiology of severe hypoxic conditions such as those for cellular oxygen delivery and demand, P/O coupling and finally control of HEP production facilitating the interaction between respiratory chain and myoglobin oxygen transport. PMID- 19807280 TI - Gonad morphogenesis in vertebrates: divergent means to a convergent end. AB - A critical element of successful sexual reproduction is the generation of sexually dimorphic adult reproductive organs, the testis and ovary, which produce functional gametes. Examination of different vertebrate species shows that the adult gonad is remarkably similar in its morphology across different phylogenetic classes. Surprisingly, however, the cellular and molecular programs employed to create similar organs are not evolutionarily conserved. We highlight the mechanisms used by different vertebrate model systems to generate the somatic architecture necessary to support gametogenesis. In addition, we examine the different vertebrate patterns of germ cell migration from their site of origin to colonize the gonad and highlight their roles in sex-specific morphogenesis. We also discuss the plasticity of the adult gonad and consider how different genetic and environmental conditions can induce transitions between testis and ovary morphology. PMID- 19807284 TI - [The influence of macro- and microstructure on the surface wettability and retention properties of endodontic posts in vitro]. AB - Aesthetical and biomechanical requirements on fiber-reinforced endodontic posts play an important role in clinical application. Leading to a long-term clinical success, post systems must ensure an adequate force transmission and optimal retention behavior into the root channel. The aim of this study was to determine the microstructure and morphology of commonly used fiber reinforced endodontic posts in vitro. Post-specific fiber diameter, fiber arrangement over the cross sections and length, and fiber-matrix ratio could be associated with mechanical properties, such as three-point bending strength and structure modulus. Furthermore, our investigation focused on the quantification of the wettability of the post surface. By using a modified contact-angle measurement, the meniscus on the post surface was characterized. In additional investigations, posts were inserted in artificial root channels and the extraction force was measured. The results show a strong correlation between the extraction force and the wettability of the post surface. PMID- 19807285 TI - [Drug release of coated dental implant neck region to improve tissue integration]. AB - In order to improve tissue integration, the neck region of dental implants was coated with the biodegradable polymer poly (L-lactide) incorporating tetracycline, ibuprofen and the combination of both drugs using a solvent dip coating process. Metallographic analysis, light microscopy and electron microscopy were used to detect the thickness range and the surface characteristics of the coatings. Cytotoxicity was evaluated using the tetrazolium colorimetric method with the fibroblast cell line L929. The in vitro drug release was measured in isotonic sodium chloride solution by UV spectroscopy. To explore if drug release is concentration-dependent, the total amount of drug was varied in the coating (20% wt, 30% wt and 40% wt). The results showed a continuous release of the embedded drugs in relevant dosage over a period of 6 months. In contrast to high tetracycline concentrations, high ibuprofen concentrations resulted in a decreased metabolic activity of the L929 fibroblasts. PMID- 19807286 TI - Smart life support: model-based design and control of life-supporting systems. PMID- 19807287 TI - Modeling and simulation of the cardiovascular system: a review of applications, methods, and potentials. AB - Proper function of the cardiovascular system is indispensible to human survival. However, this system is dominated by complex interactions between different physiological processes and control mechanisms. A structured analysis and a mathematical description of this system can provide more insight, and a computer based simulation of dynamic processes in the cardiovascular system could be applied in numerous tasks. This article gives a review of different approaches to cardio-circulatory modeling and discusses methodological aspects and fields of application for several classes of models. PMID- 19807288 TI - [Kidney and liver support therapies: state-of-the-art methods]. AB - Today, worldwide approximately 1.5 million uremic patients owe their lives to hemodialysis. This figure will increase further by a growth rate of 6% annually reflecting a necessary increasing high interest in innovative technologies as well as in individualized therapies in this area. The main focus of developmental efforts, today and in the future, is to adapt the therapy to the specific physiological conditions of kidney patients, e.g., by means of a precise control of body water and its removal. This can be precisely monitored by bioimpedance analyses. Liver patients also benefit from the experiences with extracorporeal blood circuits. In contrast to hemodialysis, protein-permeable membranes are applied which allow for the removal of albumin-bound toxins. Albumin is then cleansed in the secondary circuit by adsorber cartridges and then readministered to the patient leading to improved recovery. This short review summarizes the state-of-the-art of blood purification methods in hemodialysis and liver support therapies. PMID- 19807289 TI - [Assisted circulation: an overview from a clinical perspective]. AB - A higher grade cardiac failure is associated with poor prognosis. In addition to medical conservative treatment and traditional cardiac surgery, in the past years different forms of an assisted circulation evolved. Short-term devices serve to bridge an acute life-threatening situation. The chosen system is dependent on the anticipated clinical course. It is possible to fall back on slightly assisting techniques up to a complete takeover of the cardiac pump function. In the case of severe cardiac failure, the question for transplantation has to be addressed because transplantation is the treatment of choice to date. For an assisted circulation in cases of chronic congestive failure, devices of different generations are available. First generation pulsatile systems are used for assistance of the left ventricle and results have been shown to be superior to medical therapy (REMATCH). With second generation continuous-flow systems, results regarding infections, thromboembolism and also quality of life appear to be further improved. Contact-free centrifugal pumps as third generation systems are in clinical evaluation. So-called "total artificial hearts" are successfully used for bridge-to-transplantation. Taken together, a graded safe treatment of cardiac failure is available today. In the near future, it could be possible to reach results similar to those of cardiac transplantation. PMID- 19807290 TI - Methods of design, simulation, and control for the development of new VAD/TAH concepts. AB - Cardiovascular diseases are a major cause of death worldwide. If medical treatments fail to restore adequate blood flow in a patient, mechanical support is needed. To date, many different types of blood pumps have been developed, but only few are clinically available. This review article describes the challenges involved in this field of research and gives an overview of the development process. Past developments as well as current and new technologies and approaches applied are summarized. Finally, a perspective for improved devices is discussed. PMID- 19807291 TI - [SmartCare: automatizing clinical guidelines]. AB - In critical care environments, important medical and economic challenges are presented by the enhancement of therapeutic quality and the reduction of therapeutic costs. For this purpose, several clinical studies have demonstrated a positive impact of the adoption of so-called clinical guidelines. Clinical guidelines represent well documented best practices in healthcare and are fundamental aspects of evidence-based medicine. However, at the bedside, such clinical guidelines remain difficult to use by clinical staff. The knowledge based technology SmartCare allows incorporation of arbitrary computerized clinical guidelines into various medical target systems. SmartCare constitutes a clinical guideline engine because it executes one or more clinical guidelines on a specific medical device. SmartCare was initially applied for the automated control of a mechanical ventilator to assist the process of weaning from a medical device. The methodology allows further applications to be implemented effectively with other medical devices and/or with other appropriate guidelines. In this paper, we report on the methodology and the resulting versatility of such a system, as well as the clinical evaluation of SmartCare/PS and its perspectives. PMID- 19807292 TI - [Automatic control and safety concepts for extracorporeal lung support]. AB - In some cases of severe acute respiratory distress syndrome, hypoxemia occurs despite optimized conservative therapy; however, extracorporeal membrane oxygenation (ECMO) can assure sufficient gas exchange. To increase safety and reliability of devices, the oxygenator design was optimized integrating new plasma-resistant composite membranes and new blood pumps are used with longer durability and reduced blood cell damage. Another approach is the use of an arterio-venous pumpless extracorporeal lung assist (pECLA) using an oxygenator with reduced pressure drop to simplify management and to avoid pump-related complications. First attempts were made to integrate basic control and safety concepts in ECMO circuits, but this does not seem to be sufficient to overcome the specific problems of ECMO (long-term use and limited supervision of the intensive care unit). The integration of sophisticated automated control and safety concepts in combination with revised ECMO circuits could allow a more reliable application of ECMO of the intensive care unit without continuous observation by a perfusionist. Easier intra- and interhospital transfer of patients with running ECMO would be another advantage. PMID- 19807293 TI - [Blood pressure control in the area of surgical interventions]. AB - For specific surgical interventions, such as aortic stent implantation, it might be temporarily necessary to decrease mean arterial pressure to rather low levels (around 40 mm Hg). Such hypotensive pressure levels are necessary to avoid intra- and postoperative intricacies. Traditionally, the drug Nitroprussidnatrium is used for this task. To adjust the correct amount of drug to reach the target pressure as fast as possible and without overshoot, the anaesthetists typically use empirical knowledge and might need several minutes until the target point is reached. In our research group, an adaptive control system was developed for this task which is able to compute and set the transient drug release automatically. For the design and testing of the adaptive control strategy, the well known Guyton model was implemented into the MATLAB/Simulink development environment. This paper describes the implementation and adaption of the Guyton model to hypotensive pressure control and provides some algorithmic details of the adaptive control strategy for automatic drug delivery in deep hypotension. The designed control system was successfully validated in animal trials (25 trials on 7 pigs). Following this, an additional controller component for increase of blood pressure with the help of the drug Noradrenalin was implemented. It is now possible to increase blood pressure to a specific value to save defined cerebral perfusion pressure for patients with craniocerebral injury. In a second pilot trial, this controller extension was tested in 10 pigs. PMID- 19807294 TI - Health economic analysis of new interventions: are we asking the right questions? PMID- 19807296 TI - Evidence-based assessment of the efficacy of esomeprazole for the healing of erosive esophagitis. AB - Gastroesophageal reflux disease is generally a lifelong illness that affects many people. Erosive esophagitis can occur in up to 65% of patients with gastroesophageal reflux disease and more seriously, esophageal stricture, Barrett's esophagus and esophageal adenocarcinoma, can develop. The severity of symptoms is not a reliable indicator of the severity of erosive esophagitis and although endoscopy is the preferred method to diagnose and grade erosive esophagitis, its routine use is not practical. Therefore, early initiation of treatment with the most effective agent for treating this disease is a practical and logical strategy. Proton pump inhibitor therapy is preferred for the treatment of gastroesophageal reflux disease. This review provides a quantitative assessment of the efficacy of esomeprazole (Nexium, AstraZeneca), the most effective agent currently available. PMID- 19807297 TI - Cost-effectiveness of letrozole in the treatment of advanced breast cancer. AB - Aromatase inhibitors are fast becoming the treatment of choice in the first-line management of hormone-sensitive patients with metastatic breast cancer, replacing tamoxifen. Although these agents appear to have clinical superiority, they are more expensive alternatives, thus requiring economic evaluation to consider their incremental value to the payer. This article reviews all published economic evaluations that compared aromatase inhibitors in the first- and second-line treatment setting. From the literature, seven separate evaluations were identified, which used different types of modeling techniques and considered the payer's perspective in three healthcare systems. The conclusion of these analyses was that letrozole (Femara, Novartis) is the most cost-effective intervention in the first-line setting, whilst being of similar cost-effectiveness to anastrozole (Arimidex, AstraZeneca) in the second-line setting. In addition, preliminary results suggest that letrozole is a cost-effective alternative in the extended adjuvant breast cancer treatment setting. PMID- 19807298 TI - Cost-effectiveness studies of diabetes prevention in high-risk patients. AB - The incidence of Type 2 diabetes is increasing annually. Impaired glucose tolerance has been described as a prediabetic state, which confers an increased risk of developing Type 2 diabetes along with its associated costly complications. Interventions targeted at individuals with impaired glucose tolerance can delay or prevent the onset of Type 2 diabetes. Several recent studies have shown that the incidence of Type 2 diabetes is reduced by lifestyle changes or pharmacological interventions. A literature search for studies investigating the costs and cost-effectiveness of diabetes prevention was performed, and the results summarized in this review. The limited health economic evidence currently published suggests that financial concerns should not be a barrier to the implementation of diabetes prevention strategies. PMID- 19807299 TI - Use of electronic quality of life applications in cancer research and clinical practice. AB - Outcome measures of cancer treatment should include measures of quality of life, in addition to survival and objective response. Electronic administration of quality of life measures has made regular monitoring of the quality of life of individual cancer patients possible and feasible. Electronic questionnaires were well accepted by patients, had good test-retest reliability and were comparable with traditional paper-and-pen methods. Several randomized studies have shown that using quality of life screening in clinical practice can facilitate patient doctor communication, without increasing consultation time. In one study, the regular assessment of quality of life resulted in benefits for some patients, who had better well-being and emotional functioning. Research is ongoing in developing computer-adaptive questionnaires to reduce patient burden and provide a more precise measurement. Further research using randomized controlled studies is necessary before the monitoring of individual patients' quality of life is recommended for routine practice. Special attention should be paid to training physicians in using the quality of life scores. PMID- 19807300 TI - HRQOL and subjective well-being: noncomplementary forms of outcome measurement. AB - This review considers some of the broad principles that concern quality of life assessment. These are discussed in relation to health-related quality of life (HRQOL) and the measurement of subjective well-being. It is argued that there are serious logical and methodological issues concerning HRQOL measurement, to the extent that the instruments may not be regarded as valid measures of life quality as this term is generally understood. It is recommended that HRQOL measurement be abandoned in favor of three separate forms of measurement as medical symptoms, subjective well-being and specific dimensions of psychological ill-being. PMID- 19807301 TI - Effect of prescription drug coverage on the cost of care to Medicare beneficiaries with asthma. AB - The objective of this review is to investigate the impact of asthma on Part A and B expenditures for enrollees in the Medicare program, and the effect of voluntary outpatient prescription drug coverage on Medicare-covered asthma expenditures. Data were drawn from the 2001 Medicare Current Beneficiary Survey Access to Care file, a nationally representative sample of the Medicare population, and includes data on individual healthcare expenditures and characteristics, such as health status. Using regression analysis, it was found that self-reported asthma increased the probability of Part A (hospital) expenditures and the level of Part B (outpatient) expenditures. Voluntary prescription drug coverage decreased Part B spending on people with self-reported asthma by 7% but increased Part B spending by 23% for those without asthma. Prescription drug coverage had no effect on the level of Part A expenditures in either those with or without self reported asthma. The author's study suggests that increases in spending associated with the new Medicare prescription drug benefit may be mitigated by reductions in spending on chronic illnesses such as asthma. As the drug benefit is designed over the next 2 years, attention may need to be focused on the identification of medications that reduce overall Medicare spending to ensure that they are covered by the drug plans. PMID- 19807302 TI - Evaluating and improving the quality of care in rheumatic disease. AB - Defining and measuring healthcare quality and examining the factors that affect it are necessary to optimize health outcomes. Tools such as quality indicators and patient-directed evaluation instruments can serve to identify targets for improvement. Although the quality of healthcare for a variety of medical conditions has received increasing attention, few have attempted to measure and improve the quality of care for patients with rheumatic illnesses. This article reviews the efforts specific to musculoskeletal disease to standardize performance measurement tools. It also discusses recent quality improvement efforts targeting both structural and process components of healthcare delivery systems for arthritis and related conditions. PMID- 19807303 TI - Value of cyclooxygenase-2 specific inhibitors in the management of osteoarthritis. AB - Osteoarthritis is the most common form of arthritis and causes substantial morbidity and healthcare expenditure worldwide. This is partly due to the gastrointestinal side effects associated with the use of nonspecific nonsteroidal anti-inflammatory drugs. Cyclooxygenase-2 specific inhibitors offer a therapeutic alternative since they may reduce gastrointestinal-related risks with a similar clinical efficacy to nonsteroidal anti-inflammatory drugs. This article provides an overview of the clinical and economic value of cyclooxygenase-2 specific inhibitors in the management of osteoarthritis. The authors' findings suggest that cyclooxygenase-2 specific inhibitors show comparable efficacy with nonspecific nonsteroidal anti-inflammatory drugs, yet have reduced rates of gastrointestinal complications. Pharmacoeconomic studies have demonstrated the cost-effectiveness of prescribing cyclooxygenase-2 specific inhibitors in certain high-risk populations. Cyclooxygenase-2 specific inhibitors provide clinical and economic benefits in patients diagnosed with osteoarthritis. Future studies should further examine the economic implications of newer cyclooxygenase-2 inhibitors and quantify the impact of pain management on patients' quality of life. PMID- 19807304 TI - Economic implications of current and future provision of English learning disability health services. AB - People with learning disabilities represent approximately 2.3% of the population. Levels of physical disability, sensory impairment and mental health difficulties provide enormous challenges to the health and social services responsible for their care. Following political initiatives, such as Valuing People, statutory frameworks implementing care standards and changes to the Mental Health Act, as well as economic implications, are significant. Pharmacotherapy for the projected treatment of Alzheimer's dementia in Down's syndrome will require major investment. There is a disproportionate impact on rural services as an urban/rural divide grows in the residential care services. Increasing movement of specialist provision from the National Health Service to the private sector, with recruitment and retention issues pulling clinicians in the same direction, is slowly changing the economics of learning disability services as we know them. PMID- 19807305 TI - Clinical and economic evaluation of gatifloxacin and levofloxacin. AB - Antibiotics represent a relatively high drug cost component for the average institution. There are numerous antibiotic agents available for many indications so the choice of agents for which to provide coverage becomes complicated. The two fluoroquinolones levofloxacin (Levaquin, Ortho-McNeil Pharmaceuticals Inc.) and gatifloxacin (Tequin, Bristol-Myers Squibb) are similar in many respects yet have enough differences to allow for debate in preferring one over the other. However, overuse of these antibiotics is leading to a growing concern of bacterial resistance. This review discusses the factors that formulary decision makers consider when contemplating coverage of these drugs and explores where population-specific information would benefit this process. Both clinical and economic issues are raised in this review. PMID- 19807306 TI - Using pharmacoeconomics for policy making: is rational decision making enhanced by applying thresholds for cost-effectiveness? PMID- 19807308 TI - Depression and quality of life in patients with coronary artery disease. AB - Depression has been shown to be a risk factor for the development of coronary artery disease, morbidity and mortality after onset of coronary disease. This article reviews recent studies investigating these links and discusses the impact of depression on quality of life in patients with coronary artery disease. Recent trials of psycho- and pharmacotherapy in this population are discussed, along with the impact of treatment on prognosis and quality of life. PMID- 19807309 TI - Quality of life after aortic valve replacement. AB - Understanding the quality of life after aortic valve replacement has become increasingly important. As aortic valve replacement numbers increase, more patients, physicians and families are affected by the subsequent quality of life. Quality of life information can inform the decision to perform aortic valve replacement and the selection of replacement valve. When reviewing quality of life results, it is important to realize that the findings are affected by the selection and quality of instruments, as many studies have not used valid or reliable instruments. Studies have shown that aortic valve replacement appears to significantly improve the quality of life of survivors, including those older than 70 years of age and even decades after the procedure, quality of life remains high. Studies have suggested that the elderly may gain as much quality of life benefit as younger patients. No consistent differences in resulting total quality of life have been observed between mechanical and bioprosthetic valves. Only one study showed some quality of life benefits of pulmonary autograft over mechanical valves. It is unclear whether minimally invasive aortic valve replacements confer better quality of life than standard aortic valve replacements. While existing quality of life studies have provided important information, more studies are needed especially as valve technology and operative techniques continue to improve. Future studies should endeavor to use validated general and disease-specific instruments and quantify the effects of demographics, preoperative clinical conditions and intraoperative variables on quality of life outcomes. PMID- 19807310 TI - Short- and long-term HRQOL issues following brachytherapy for prostate cancer. AB - Given the lack of evidence from randomized controlled trials and large prospective studies, controversy exists regarding the optimal treatment strategy for early stage prostate cancer. The impact of various treatments on health related quality of life has become an increasingly important but controversial issue. The literature on health-related quality of life following brachytherapy, an increasingly used treatment option that can have short- and long-term side effects influencing patient health-related quality of life is reviewed. Urinary, bowel and sexual health-related quality of life issues related to this treatment are addressed. The authors findings suggest that brachytherapy with or without external beam radiotherapy and/or hormonal treatment has a considerable impact on short-term health-related quality of life but this impact diminishes in the long term. However, in certain subgroups of patients, brachytherapy has long-term effects on bowel, bladder and sexual function. Clinicians should ensure that patients are aware of these consequences on long-term health-related quality of life. PMID- 19807311 TI - Cost-effectiveness of cervical cancer screening strategies. AB - Despite its low incidence in the developed world, cervical cancer is the second most common cancer among women worldwide and the third most common cause of cancer-related deaths. Screening saves lives. When diagnosed and treated in the preinvasive stage, 5-year survival probabilities approach 100%. This review addresses the cost-effectiveness of the various screening strategies commonly used worldwide. The starting age of screening, interval between screens and age of women at cessation of screening are the most important variables determining cost-effectiveness. While the traditional Pap test is still the most cost effective screening method in most settings, human papillomavirus testing can improve outcomes at reasonable costs in certain high-risk populations. PMID- 19807312 TI - Cost-effective therapy in patients with idiopathic hirsutism. AB - Hirsutism affects 10% of women. Hirsute women with normal circulating androgen levels and normal ovarian function (i.e., regular and ovulatory menstrual cycles) are defined as having idiopathic hirsutism, which may affect more than 20% of all hirsute women. In the treatment of idiopathic hirsutism, different medical therapies, alone or in combination, have been reported. The drugs currently available are oral contraceptives, cyproterone acetate androgen receptors blockers (i.e., spironolactone and flutamide), 5alpha-reductase inhibitors (e.g., finasteride [Proscar, Aventis]) and gonadotrophin-releasing hormone analogs. After 1 year of treatment, each drug may improve hirsutism and reduce the Ferriman-Gallwey score by 35-40%. This review analyses the causes of hirsutism and provides information on each therapy and the cost-effective results in patients with idiopathic hirsutism. PMID- 19807313 TI - Cost-effectiveness analysis of biological treatments for rheumatoid arthritis. AB - This review compares the cost-effectiveness of four biologics - adalimumab (Humira, Abbott Laboratories), anakinra (Kineret, Amgen Inc.), etanercept (Enbrel, Wyeth) and infliximab (Remicade, Schering-Plough) - used in the treatment of rheumatoid arthritis. A decision analytic model was constructed to estimate the costs and effectiveness of these biologics used alone or in combination with methotrexate during 1 year, from the perspective of a managed care organization. The direct costs consisted of drugs and healthcare resources. Effectiveness was measured by quality-adjusted life years based on preference weights and health states in which patients achieved one out of four levels of response according to the American College of Rheumatology (ACR) response criteria (No ACR, ACR20, ACR50 and ACR70), and experienced one of the four levels of adverse events (e.g., no, mild, moderate and severe) due to their treatments. Results were sensitive to changes in treatment costs and probabilities of health states in directions as predicted. For monotherapy and combination therapy regimens, anakinra was the least expensive option while etanercept dominated other treatments. PMID- 19807314 TI - Different aspects of adherence to antihypertensive treatments. AB - This review collects together data concerning the measurement of patient behavior towards antihypertensives, considering all its aspects - adherence, persistence and switching. The wide heterogeneity of terms, outcome definitions and methods of assessment (the multiplicity of approaches did not allow a full meta-analysis, but a qualitative description of the results) are demonstrated. The percentages of adherence/compliance from the studies discussed in this review range from 15 to 99% across different study designs, populations and methods. Large, observational studies are suggested as a reliable method to measure patient behavior. An emerging issue is the relevance of patient motivation and education, which could provide a higher level of compliance to treatment, thus allowing achievement of better clinical outcomes and improved use of resources for the healthcare system. PMID- 19807315 TI - Effects of introducing a clinical guideline on the cost of treating dyspeptic syndrome. AB - This review highlights the effects incurred on pharmacoutilization and the total expenses for dyspeptic syndrome by the introduction of a clinical guideline. A retrospective method of reading an administrative billing database in the Ravenna (Italy) local health unit was performed for all subjects who were health-assisted by ten general practitioners. They had previously developed and agreed to a clinical guideline to manage dyspeptic syndrome patients (study group) and a group of 30 self-regulating general practitioners patients (control group). Patients were classified as dyspeptic according to the presence of at least one prescription for antidyspeptic drugs during a 365-day follow-up period. The cost for drugs, hospitalizations, emergency room accesses and gastroscopies was recorded. The results showed that a total of 51,904 subjects were enrolled: 23.1% by the study general practitioners and 76.9% by the control general practitioners. The percentage of dyspeptic patients accounted for 17.6 and 15.0% in the subjects enrolled in the study and control groups. The total average cost of the dyspeptic patients was less in the study than in the control group (euro163.41 vs. 181.39; p = 0.043). This difference is mainly due to the cost of hospitalization (euro94.55 in the study and euro110.92 in the control group; p = 0.012). The significant results of the introduction of a guideline for the treatment of dyspeptic syndrome into clinical practice were an increase in the use of pharmacological treatment and a decrease in the cost of treatment per patient. PMID- 19807316 TI - Changes in drug spending for different age groups during the 1990s? Evidence from Sweden. AB - The objective of this review is to examine how drug spending in different age groups changed during the 1990s. Time series analysis of registered data on prescription drug spending were performed, along with two decompositions, one of which was spending in three components: price, quantity (defined daily dose) and residual. The size of the residual is a measure of the impact of changes in drug treatment patterns on drug spending. The other decomposition was of the quantity component in three subcomponents: defined daily doses per person on medication, population share on medication and population size. Both decompositions are made separately for different age groups. How spending for different age groups has developed in different therapeutic areas was also studied. The main outcome measures were prescription drug spending over time within different anatomical therapeutic chemical groups and across different age groups of the population. It was found that the older the age group, the more drug spending had increased, both in absolute and in relative terms, during the 1990s. However, for some anatomical therapeutic chemical groups, younger age groups have experienced faster spending growth. The most notable example being anatomical therapeutic chemical group N CNS, where spending grew fastest by 350%, for those aged between 20 and 39 years. Furthermore, changed treatment patterns, such as a switch to more expensive drugs, is the main explanation for higher spending in all age groups. Higher spending is also due to a larger number of defined daily doses sold, which is almost totally due to the fact that each person on medication in the year 2000 utilized more defined daily doses than in 1990. Changing age structure explains a negligible share of the increase in drug spending, but elderly patients did have a key role in the spending surge since they increased their per capita spending the most. PMID- 19807317 TI - Burden of headache. AB - Although headache is the most prevalent pain disorder, the substantial societal and individual burden associated with it has been previously overlooked. The primary headaches including migraine, tension-type and cluster are the most prevalent forms but also a very high number of headaches secondary to analgesic overuse and various brain diseases are ignored without any specific acknowledgement or treatment. Migraine is listed as number 20 of all diseases with regard to years lived with disability, and headache accounts for approximately 20% of lost work days. Chronic headaches (occurring for more than half of all days per month) affects 4-5% of the general population and differs not only from the episodic forms in frequency, but also in respect to lack of effect of most treatment strategies, more medication overuse and more loss of quality of life than the episodic forms. The burden of headache disorders remains substantial and is estimated to cost a minimum of US$100 million per million inhabitants per year. As most headache disorders remain prevalent, disabling, underdiagnosed and undertreated, public health initiatives to improve prevention and treatment are urgently required. Considerable benefits for society can be gained by specific strategies leading to reductions in the amount of sickness absence and impaired working abilities. The burden on affected individuals and their families quality of life may be improved by a general acceptance of the impact of headache disorders and the development of specific treatment strategies. PMID- 19807318 TI - Health economic consequences of an aging population. PMID- 19807319 TI - Quality of life in oncology clinical trials: present and future challenges. PMID- 19807321 TI - Probabilistic sensitivity analysis for evaluating cost-utility of entacapone for Parkinson's disease. AB - The objective was to assess uncertainty in a cost-utility analysis of adjunct entacapone treatment with levodopa among Parkinson's disease patients by probabilistic sensitivity analysis using second-order simulation methods. The cost-effectiveness of two treatment alternatives of Parkinson's disease - levodopa with or without entacapone - was compared in a cost-utility analysis employing a Markov model. Monte Carlo simulation was used to quantify the uncertainty due to sampling variation. The results strengthened confidence in the conclusions that entacapone as an adjunctive treatment to levodopa is both cost saving and increases the quality of life of Parkinson's disease patients. PMID- 19807322 TI - Health utility estimation. AB - Preference measurement techniques, like the standard gamble, time trade-off, visual analog scale, EuroQol, Health Utilities Index and Quality of Well-Being, are increasingly appearing in published studies. The scores from these instruments are variously called utilities, values or preferences. What are these concepts and do they differ? What are these instruments and what do they measure? What does a reader need to know to make sense of it all? What does a user need to know to select an appropriate approach? Read on for the answers. PMID- 19807323 TI - Measuring health-related quality of life in the clinical setting. AB - Quality of life is recognized as an important health outcome and several measures of quality of life have been well validated for use in clinical trials and outcomes research. These measures could play an important role in routine clinical care, providing valuable information on physical functioning, emotional health and other outcomes of interest to patients and providers alike. The literature to date largely reports on how these measures might be used in clinical practice, with few published studies on actual experiences with their implementation. Barriers to the use of quality of life measures, such as cost and time of administration, may decrease thanks to a new generation of measurement methodologies and tools. PMID- 19807324 TI - Methodological issues affecting the value of patient-reported outcomes data. AB - The validity and value of patient-reported outcomes data are heavily dependent on the methods used to collect the data. This review examines the impact of a variety of methodological issues on the value of patient-reported outcome data. In particular, when patients are asked to self-monitor their experiences, disease episodes and healthcare utilization over time, a variety of methodological issues must be addressed if the data are to be considered a reliable and valid reflection of their daily lives. Ecological momentary assessment, a set of methods for collecting real-time data from patients in their natural environments, holds considerable promise as a way to enhance the value of certain types of patient-reported outcome data. PMID- 19807325 TI - Quality of life issues following autologous bone marrow transplantation. AB - Over the past decade, the use of autologous bone marrow or stem cell transplantation has become common in the treatment of hematological malignancies and some solid tumors. However, little was known about the effects of these procedures on QoL outcomes, such as physical, emotional, social and role functioning. Recent research allows identification of the most common QoL issues in this group of patients, both during acute treatment and through long-term follow-up. This paper systematically summarizes all published articles that have investigated QoL in autologous bone marrow or stem cell transplantation patients, including comparisons with patients undergoing allogeneic bone marrow transplant, chemotherapy alone and normal control subjects. Several consistent QoL problems are identified and suggestions for treatment and further research explored. PMID- 19807326 TI - Psychosocial factors in cardiovascular disease: implications for therapeutic outcomes. AB - The etiology of cardiovascular diseases is complex, involving the interaction of genetic, biologic, behavioral and psychosocial factors. The role played by psychosocial factors in the etiology and progression of cardiovascular diseases stems from several sources: their influence on behaviors that either promote health or increase risk, the neuroendocrine changes accompanying emotions and their interaction with genetic factors. The focus of this article is the elucidation of pathways that mediate the effects of psychosocial factors on cardiovascular health and the implications for clinical intervention. PMID- 19807327 TI - Long-term health outcomes and mechanisms associated with breastfeeding. AB - Breastfeeding is superior to formula feeding because it has factors that have long term consequences for early metabolism and disease later in life. In this paper, the scientific evidence in support of why breast milk is beneficial for infants is summarized and the mechanisms in which breastfeeding impacts on disease are explored. Human milk may show a reduced occurrence of disease because mammalian evolution promotes survival, and because of specific factors in milk that promote active stimulation of the infant's immune system and gastrointestinal mucosal maturation decrease the incidence of infection and alter the gut microflora. Bioactive factors, including: hormones, growth factors, colony-stimulating factors and specific nutrients, may have such far-reaching effects on the infant's immune response that normal development depends heavily on its provision. All mothers should be encouraged and supported to continue breastfeeding for 6 months and beyond in order to promote the good health of their infants. PMID- 19807328 TI - Decision analysis and drug development portfolio management: uncovering the real options value of your projects. AB - Project selection and portfolio management are particularly challenging in the pharmaceutical industry due to the high risk - high stake nature of the drug development process. In the recent years, scholars and industry experts have agreed that traditional Net-Present-Value evaluation of the projects fails to capture the value of managerial flexibility, and encouraged adopting a real options approach to recover the missed value. In this paper, we take a closer look at the drug development process and at the indices currently used to rank projects. We discuss the economic value of information and of real options arising in drug development and present decision analysis as an ideal framework for the implementation of real options valuation. PMID- 19807329 TI - Leisure time in economic evaluation: theoretical and practical considerations. PMID- 19807331 TI - Cost-effectiveness and HRQOL outcomes of divalproex sodium (Depakote) in bipolar disorder. AB - Bipolar disorder is a chronic and severe affective disorder which has a significant impact on patient functioning and well-being, and on total medical costs. Divalproex sodium (Depakote) has been shown to be effective in treating acute mania and for the long-term treatment of bipolar disorder. Few health related quality of life or pharmacoeconomic studies have been completed comparing divalproex sodium with other mood stabilizers. Clinical trials have demonstrated that the response to divalproex sodium therapy is associated with improvements in health-related quality of life outcomes and reduced disability days. No statistically significant differences on health-related quality of life outcomes have been observed between divalproex sodium, lithium or olanzapine (Zyprexa) treatment in bipolar disorder. Effective mood stabilizer therapy results in improvements in patient functioning and well-being, but there is little evidence differentiating the effects of divalproex sodium on these outcomes compared with other mood stabilizer treatments. The current pharmacoeconomic studies suggest that divalproex sodium is associated with lower total and inpatient medical costs compared with lithium. A small, recent clinical trial has also demonstrated that total outpatient costs are lower for divalproex sodium compared with olanzapine. Additional cost-effectiveness studies are required to examine the impact of divalproex sodium versus other mood stabilizers in the treatment of bipolar disorder. PMID- 19807332 TI - Drotrecogin alfa (activated; Xigris): an effective and cost-efficient treatment for severe sepsis. AB - Severe sepsis is a common health problem with consequences for both patients and the healthcare system. Over the past 20 years, multiple immunomodulatory agents have been investigated in an unsuccessful attempt to decrease the morbidity and mortality of severe sepsis. Drotrecogin alfa (activated; Xigris) may represent a breakthrough in the treatment of sepsis. It has been demonstrated to have beneficial effects in decreasing biological markers of the severity of sepsis in preclinical and Phase II studies. A single, large Phase III trial has demonstrated the efficacy of drotrecogin alfa (activated) in a sample of patients with severe sepsis. This sample appears to be comparable with the general population of patients with severe sepsis. Three separate economic analyses have shown drotrecogin alfa (activated) to have a cost-utility ratio similar to other therapies that are currently funded, when used for the treatment of the most severely ill group of patients. This review provides an opinion that drotrecogin alfa (activated) is a cost-efficient therapy that should be considered as part of a standard of care in healthcare systems that can provide a modern critical care service. PMID- 19807333 TI - Cost-utility analysis of the GC versus MVAC regimens for the treatment of locally advanced or metastatic bladder cancer. AB - A cost-utility analysis was conducted to compare the gemcitabine (Gemzar)/cisplatin and methotrexate/vinblastine(Velban)/doxorubicin (Doxil)/cisplatin regimens in the treatment of locally advanced or metastatic bladder cancer. The analysis combined the results of a cost analysis of a head-to head Phase III clinical trial with those from a cross-sectional utility study. The analysis was performed from the perspective of the National Health Service in England and Wales. Medical resource use was obtained from the clinical trial database, from which patient-level cost streams were estimated for each treatment group. Unit costs for each resource utilization component were sought from the latest UK cost sources available at the time of analysis (2001). The distribution of the mean total and incremental costs for each group was simulated via bias adjusted bootstrapping. Time trade-off utilities were derived from the utility study, which aimed to value the superior toxicity profile associated with gemcitabine/cisplatin, given comparable efficacy, in a discrete choice model assessing toxicity attributes and risks. The mean incremental cost of gemcitabine/cisplatin over methotrexate/vinblastine/doxorubicin/cisplatin was estimated to be approximately pound sterling 2976 per patient, based on a mean of 4.65 cycles per patient treated with gemcitabine/cisplatin compared with a mean of 3.92 cycles per methotrexate/vinblastine/doxorubicin/cisplatin patient. When combined with the utility estimates, this resulted in an incremental cost effectiveness ratio of approximately pound sterling 22,925 per quality-adjusted life year gained associated with the choice of gemcitabine/cisplatin over methotrexate/vinblastine/doxorubicin/cisplatin. A 95% confidence interval for the incremental cost-effectiveness ratio was calculated to range from pound sterling 12,911 to 33,589 per quality-adjusted life year gained. The incremental cost of gemcitabine/cisplatin is primarily due to the direct costs of chemotherapy given minimal cost offsets. However, an incremental cost per quality-adjusted life year ratio in the range of pound sterling 20,000-30,000 per quality-adjusted life year gained suggested that the regimen is reasonable value for money in England and Wales. PMID- 19807334 TI - Economic assessments of low molecular weight heparin in venous thromboembolism. AB - Heparin is the treatment of choice for managing patients in the acute phase of venous thromboembolism. While the drug cost of low molecular weight heparins may be greater than unfractionated heparin, its use makes outpatient treatment possible for most patients and allows an earlier discharge from hospital for those requiring inpatient care. Pharmacoeconomic studies and meta-analyses have compared disease management with unfractionated heparin versus the low molecular weight heparins. The results indicate that low molecular weight heparins are the dominant choice. Health outcomes are improved while the overall cost of managing venous thromboembolism is reduced. Additional research is required to examine the relative efficacy and safety of each of the low molecular weight heparin presentations, adherence in the outpatient setting and the impact of acute treatment choices on long-term outcomes such as recurrence, post-thrombotic syndrome and quality of life. PMID- 19807335 TI - Pharmacoeconomic review of recombinant human DNase in the management of cystic fibrosis. AB - For the treatment of patients with cystic fibrosis, recombinant human deoxyribonuclease I is widely used. Deoxyribonuclease I has a positive effect on lung function and the number of hospitalizations. Deoxyribonuclease I is currently administered by nebulization, which is an inefficient administration method. For expensive drugs, such as deoxyribonuclease I, dry powder inhalation would be advantageous due to increased deposition efficiency, patient mobility and compliance. Furthermore, a significant cost reduction may be obtained. The current status of deoxyribonuclease I in the management of cystic fibrosis was investigated and special attention given to the developments in delivery systems, such as dry powder inhalation. It is estimated that if dry powder inhalation of deoxyribonuclease I could be used, a reduction in the cost-effectiveness ratio of approximately 40% can be obtained as compared with nebulization. PMID- 19807336 TI - Economics of cancer screening programs. AB - Cancer is a common cause of mortality in the industrialized world. Several forms of the disease are preventable by screening and mass population screening programs have been implemented in many countries. Given the current concern over value for money in healthcare, cancer screening has increasingly attracted the attention of economists. In this review, it is argued that obtaining evidence to demonstrate cost-effectiveness with respect to a novel screening hypothesis is expensive, difficult and time-consuming, with the result that, in the past, screening programs have been implemented on the basis of inadequate evaluation evidence. Over-enthusiastic implementation has led to the institutionalization of inefficient programs, although the tendency to implement too rapidly can be explained by a simple economic model of incentives. Unfortunately, once programs have become established, the economic justification for screening becomes even more difficult to prove. PMID- 19807337 TI - Self-management interventions to improve outcomes in patients suffering from COPD. AB - New research and evidence-based self-management programs are now emerging in chronic obstructive pulmonary disease. Patients with moderate-to-severe chronic obstructive pulmonary disease who receive a self-management education with supervision and support of a case manager, have better outcomes than patients receiving standard care. The benefits are on patients' health status and healthcare utilization with markedly decreased hospitalizations, emergency department and unscheduled physician visits. These benefits are worth considering since self-management could also be cost saving. There is no reason to question the value of self-management in chronic obstructive pulmonary disease, especially for those patients who have a decreased health-related quality of life and high exacerbation rate. Self-management should be an integral part of the long-term care of chronic obstructive pulmonary disease patients. Rather than telling the patient to learn to live with their disease, they should be taught to self manage. PMID- 19807338 TI - Cost-effectiveness of managing HIV infection. AB - The worldwide spread of HIV infection continues, particularly in economically vulnerable, developing and emerging countries. There is a lack of resources to supply effective prevention measures and apply highly active antiretroviral therapies. In the regions concerned, economical effects of morbidity- and mortality-associated productivity losses in those of a working age remain serious. Thus, the United Nations Program on HIV/AIDS and the World Health Organization have predicted a dramatic decrease of South Africa's and other African states' domestic product in the coming years. Therefore, with the world economical consequences from the HIV pandemic, a large political challenge in the next 10 years will arise. PMID- 19807339 TI - Update on statistical power and sample size assessments for cost-effectiveness studies. AB - Traditionally, competing healthcare interventions have been compared on their efficacy or effectiveness through clinical trials or epidemiological studies. Computation of sample size and/or statistical power for a proposed study to assess the relative effectiveness of two treatments is central to planning a good study. Economic evaluation studies focus on an additional dimension by comparing these interventions with respect to their cost. With rising healthcare costs and constrained budgets, these studies are increasingly being performed to ascertain which interventions can deliver additional health benefits at a reasonable cost. The design of a cost-effectiveness study for two competing treatments will require assessments of statistical power and sample size in demonstrating both effectiveness and/or cost-effectiveness. If the level of effectiveness of one treatment is known from a clinical trial, the next step is to assess its cost effectiveness. In other circumstances, an investigator may wish to design a study that simultaneously assesses both effectiveness and cost-effectiveness. Statistical methodologies that have been proposed to address these issues are reviewed. PMID- 19807340 TI - Nursing home costs and quality of care: is there a tradeoff? AB - Nursing homes are facing increased environmental pressures to improve quality of care. However, this increased emphasis on quality comes at a time when nursing homes are facing revenue constraints as a result of the repeal of the Boren amendment, increased reimbursement based on prospective payment systems and widespread cuts to state Medicaid funding. This represents a strong financial incentive to control costs and there are concerns that these cost-containment measures may lead to a lower quality of care in nursing homes. In this article, the scientific literature on the cost-quality relationship in the nursing home industry is reviewed. PMID- 19807342 TI - Pharmacoeconomic studies and the pharmaceutical industry: bias in which direction? PMID- 19807341 TI - Thriving after hematopoietic stem cell transplant: a focus on positive changes in quality of life. AB - Studies describing the impact of hematopoietic stem cell transplants on patient quality of life have burgeoned in the last several years. The picture of the negative impact of a hematopoietic stem cell transplant on both the short- and long-term quality of life and the extent to which patients return to baseline quality of life functioning is almost complete. However, studies of positive changes in the form of gains or growth are lacking. Reports of psychological growth or benefit-finding have been reported in the context of other significant stressors including cancer. Yet, few studies have examined similar gains experienced by patients and families after hematopoietic stem cell transplant. These studies are reviewed and the authors findings are described, making note of conceptual and methodological challenges in this area. Understanding positive changes and gains in posthematopoietic stem cell transplant provides critical knowledge to inform practice guidelines and interventions designed to both minimize negative and maximize positive outcomes. PMID- 19807344 TI - Pharmacoeconomic aspects of memantine (Ebixa) in the treatment of Alzheimer's disease. AB - Severe dementia is a devastating condition causing great impairment in cognition and functional capacity. The costs associated with severe dementia are very high and the disease can have a major impact on the next of kin. Thus, any treatment that can influence the course of severe dementia would be of great value. Memantine (1amino3,5dimethyladamantane hydrochloride, CAS 4100521, Ebixa) is a noncompetitive N-methyl-D-aspartate antagonist. It has been suggested that by inhibiting excessive stimulation of NMDA receptors, memantine may have the potential to provide both symptomatic improvement and neuroprotective effects in Alzheimer's disease. In clinical trials on patients with moderate-to-severe Alzheimer's disease, memantine has shown significant effects on clinical global impression of change, behavior and activities in daily life. In this survey, pharmacoeconomic results of memantine treatment from a prospective randomized 6 month US clinical trial are reviewed. Caregiver time was significantly lower in the memantine group and the resulting costs were also lower in the memantine group as compared with placebo. Combined with efficacy results, this may indicate cost-effectiveness. However, such a conclusion should be judged with some care since the long-term effects on resource utilization, costs and survival are not known. PMID- 19807345 TI - Self-management programs in chronic obstructive pulmonary disease: are they worthy? AB - Chronic obstructive pulmonary disease is a smoking-related progressive disease with an increasing prevalence and socioeconomic burden worldwide. Educational programs are usually components of rehabilitative strategies. Current intervention is through educational programs designed for chronic obstructive pulmonary disease patients. Among these programs, patient education on sustained smoking cessation was shown to be the only approach able to interfere with the natural history of this disease. The self-management model approach in chronic obstructive pulmonary disease is similar to the interventions initially applied in asthma. Previous data suggested that educational intervention did not influence health-related quality of life but enabled patients to learn when to adequately self-administer oral steroid and antibiotic treatment. The evaluated study is aimed at assessing the impact of a self-management strategy on health related quality of life and health resource utilization. PMID- 19807346 TI - Usefulness of resource utilization estimates from piggyback studies in rheumatoid arthritis. AB - Several methods to derive estimates of resource utilization for pharmacoeconomic evaluations exist. A common method is to collect this information alongside a clinical trial. A common criticism of this data collection strategy is that it may yield inaccurate results as the clinical trial protocol places many restrictions on the study design, patient enrolment and frequency of data collection, which may lead to an artificial picture of resource consumption and costs. In this special report, this problem is examined in rheumatoid arthritis patients. Data were collected from two sources: a Phase III clinical trial with piggyback resource utilization and a cross-sectional database utilizing retrospective recall of resource use. The patients from each database were matched in terms of age, gender and Health Assessment Questionnaire Disability Index and major measures of resource use were compared. The results of this study suggest that there were limitations in the use of patient-reported data to estimate resource utilization in this clinical trial. PMID- 19807347 TI - Impact of socioeconomic status on cardiovascular outcomes in Canada. AB - Socioeconomic status is an important predictor of cardiovascular health and outcomes both in Canada and worldwide. Social epidemiologists have struggled to identify the causal pathways that link poverty with death. While some have hypothesized that socioeconomic-outcome disparities are merely an epiphenomenon of analytic pitfalls, others perceive that wealth-health gradients are attributable to variations in patient biology, disparities in care or both. In this review, the background literature surrounding socioeconomic-outcome gradients in cardiovascular disease are briefly summarized. The Socioeconomic and Acute Myocardial Infarction study, has been designed to further understand the intermediary pathways which link socioeconomic status to outcomes following acute myocardial infarction. PMID- 19807348 TI - Measurement strategies for indirect costs in economic evaluations. AB - Considerable research has been undertaken on the different methods for valuing work absences and productivity losses (e.g., human capital vs. friction costing). However, investigation related to the best way to capture this information directly from trial participants or through surveys has been inadequate. The collection of information that may be used in providing estimates of indirect costs is problematic for a number of reasons such as the presence of poor questionnaire design, a lack of psychometric testing and accessible validating data and the potential for recall errors. In this review, the measurement methods of lost time for work activities are examined. Issues in the validity of existing instruments and recall periods are explored. Based on a review of the literature, the relevance of generic and specific methods of measuring work losses are assessed. PMID- 19807349 TI - Methodological issues in the monetary valuation of benefits in healthcare. AB - Economic evaluation is concerned with comparing the costs and benefits of competing healthcare interventions in order to efficiently allocate resources. Whilst valuing costs is important, the valuation of benefits represents one of the greatest challenges facing health economists. This review considers the different techniques available for the valuation of healthcare benefits in monetary terms, and more specifically the contingent valuation method and discrete choice experiments. This review identifies some of the key issues and debates that have been published in the literature concerning the application of the techniques to healthcare and highlights important areas for future research. PMID- 19807350 TI - Economic considerations and future directions for lung volume reduction surgery. AB - Chronic obstructive pulmonary disease is a major cause of morbidity and mortality throughout the world. Standard management includes avoidance of causative factors such as smoking, and pharmacological therapy with combinations of inhaled and/or oral bronchodilators and corticosteroids. Lung transplantation is an established but infrequent option due to donor shortages. The nonevidence-based application of lung volume reduction surgery as palliation for patients with severe emphysema culminated in the US federally approved multicenter National Emphysema Treatment Trial published in 2003. This article reviews recent guidelines concerning chronic obstructive pulmonary disease management as outlined in the 2002 Global Initiative for Obstructive Lung Disease Workshop Report, focuses on published data regarding the cost-effectiveness of lung volume reduction surgery and outlines some less invasive techniques that are on the horizon. PMID- 19807351 TI - Cost-effectiveness of Advair: fluticasone propionate and salmeterol combination. AB - The National Heart, Lung and Blood Institute guidelines for the treatment of asthma suggest that inhaled corticosteroids, with the addition of a long-acting bronchodilator, may be the most effective long-term control medication for asthma. Five inhaled corticosteroids are approved for use in the USA, including beclomethasone dipropionate, budesonide, flunisolide, triamcinolone acetonide and fluticasone propionate. Fluticasone propionate (Flixotide) and the long-acting beta2 agonist salmeterol (Serevent), are now available in the USA together in an easy to use dry powder inhaler Advair. The cost-effectiveness of this combination in the treatment of persistent asthma is reviewed. This review evaluates all the cost-effectiveness studies comparing fluticasone propionate and salmeterol from two separate inhalers or one single inhaler available in the literature. Cost effectiveness was compared with inhaled corticosteroids alone, leukotriene receptor antagonists and other combination therapies in patients with persistent asthma. PMID- 19807352 TI - Cost-effectiveness of community-acquired pneumonia therapy. AB - Community-acquired pneumonia is a common disease in adults and substantially contributes to morbidity and mortality in the USA and worldwide. Due to the significant costs associated with this disease, there is increasing pressure to evaluate the variation in practices among healthcare providers. The processes of care related to the diagnosis, management and prevention of community-acquired pneumonia are reviewed. Furthermore, the cost-effective strategies for community acquired pneumonia and the medical evidence that support their usage are outlined. PMID- 19807353 TI - Potential cost-effectiveness of colorectal cancer chemoprevention with nonsteroidal anti-inflammatory drugs. AB - Colorectal cancer is the second leading cause of cancer-related death in the Western world. Screening for colorectal neoplasia, including the removal of adenomas, is highly effective and cost-effective in reducing colorectal cancer incidence and mortality. However, only a minority of the population is currently screened. Based on data from animal models, observational studies and randomized trials in humans, nonsteroidal anti-inflammatory drugs appear to have great promise as chemopreventive agents against colorectal cancer. The critical factors that will determine the roles of aspirin, other nonselective nonsteroidal anti inflammatory drugs and selective cyclooxygenase-2 inhibitors in colorectal cancer chemoprevention include the magnitude of their protective effect, their risks, their costs, the treated population's characteristics, treatment adherence rates and how chemoprevention compares with established screening strategies. PMID- 19807354 TI - Economic evaluation of aromatase inhibitors for the treatment of breast cancer. AB - Breast cancer affects 1 million women a year. Hormonal therapy has been a mainstay of treatment for women with hormone-sensitive breast cancer. The newer class of aromatase inhibitors has challenged the use of the previous standard agent, tamoxifen, in the treatment and prevention of breast cancer. This article will provide a thorough review of the literature on the newer generation aromatase inhibitors and examine their role in metastatic, adjuvant, neoadjuvant and prevention settings. An in-depth review of the cost-efficiency analyses is also performed. Recent studies on the newer, third-generation aromatase inhibitors have challenged the previous gold standard (tamoxifen) in the prevention and management of hormone-sensitive breast cancer in postmenopausal women. These studies have supported the use of aromatase inhibitors as first-line therapy in both early (adjuvant) and advanced breast cancer management. Whether aromatase inhibitors should replace tamoxifen in these settings is still a matter of debate. Augmenting these studies and providing additional information to the debate are pharmacoeconomic appraisals, which help to place a societal value on newer therapies as compared with the previously established therapy. This review discusses the pharmacology and efficacy of the newer aromatase inhibitors as it applies to their role in the metastatic, adjuvant, neoadjuvant and prevention settings. The pharmacoeconomic studies performed on these agents are also discussed. PMID- 19807355 TI - Item response theory and health-related quality of life in cancer. AB - Identifying health-related quality of life concerns is a priority when caring for people with cancer. Specific problem areas such as pain, fatigue, emotional distress, disease- and treatment-related symptoms, as well as physical functioning can be routinely assessed using applications that draw upon item response theory. Item response theory measurement models can improve on the classical approach to health-related quality of life assessment with advantages that include comparison of patients across diverse instruments, flexibility in degree of precision desired, availability of multiple short forms, interval measurement and capability for individual assessment (real-time clinical monitoring) using computerized adaptive testing. This review describes a model of health-related quality of life in oncology and the contribution of item response theory to assessment using that model. PMID- 19807356 TI - Societal impact of cocaine-associated chest pain: when truth confronts the'great white lie'. PMID- 19807358 TI - Economic analysis of a self-management asthma plan: is it really effective if further assessed? AB - Bronchial asthma is a disease with an increasing socioeconomic burden worldwide. New strategies are currently trying to reduce the burden by the means of implementing cost-effective management guidelines. The most commonly used is the so-called guided self management plan, so that patients are taught to adjust their treatment according to changes in the severity of their disease and the existing therapeutic guidelines. Such plans proved their effectiveness in reducing asthma-related morbidity, but little is known about the real costs of implementing them. The study authored by Schermer TR and colleagues comparatively assessed the cost-effectiveness of a self-management asthma plan versus usual care applied at primary care for a period of 2 years and found that the former added more successfully treated weeks and quality-adjusted life years than the latter. PMID- 19807359 TI - Salmeterol/fluticasone combination in COPD: is together better? AB - Chronic obstructive pulmonary disease represents one of the chronic conditions for which many therapies have become available during the last few decades. They are aimed at reducing symptoms such as dyspnea, cough and sputum and many of them are given on inhalatory route. Among them, bronchodilators, such as; salmeterol or formoterol and inhaled steroids, such as budesonide or fluticasone have each proven their effectiveness in improving many clinical outcomes such as dyspnea, exacerbations number, lung function decline or health status. Currently combinations of these two classes are available in an attempt to make the inhalatory therapy more effective. These studies analyse the effectiveness of salmeterol/fluticasone combination from a clinical point of view and for a short period of time. PMID- 19807360 TI - Incorporation of environmental factors into outcomes research. AB - In health and disability arenas, it is increasingly being recognized that removing or modifying environmental factors can have a greater influence over outcomes than many individually focused interventions. In 2001, the World Health Organization endorsed a major revision of its framework for assessing and classifying health, disability and handicap, conceptualizing intervention and assessing outcome. This framework, the International Classification of Functioning, Disability and Health (ICF), is now defined by its recognition of the impact of environmental and personal factors on body function and structure, activities and participation in disablement. The ICF offers the potential to advance the understanding and integration of environmental dimensions into outcome research and measurement in health and disability. This paper proposes that a key future challenge for outcomes research is to understand and document environmental dimensions of health and disability using the precedent of the ICF. Potential steps and obstacles to this development are suggested, and the direct practice and broader policy applications gained by linking an international conceptual framework with clinical outcome research and practice are discussed. PMID- 19807361 TI - Item response theory modeling in health outcomes measurement. AB - There is a great need in health outcomes research to develop instruments that accurately measure a person's health status with minimal response burden. This need for psychometrically sound and clinically meaningful measures calls for better analytical tools beyond the methods available from traditional measurement theory. Applications of item response theory (IRT) modeling have increased considerably because of its utility for instrument development and evaluation, scale scoring, assessment of cultural equivalence, instrument linking and computerized adaptive testing. IRT models the relationship between a person's response to a survey question and their standing on a health construct, such as fatigue or depression. This review will discuss the theory and basics of IRT models and applications of these models to health outcomes measurement. PMID- 19807363 TI - Pharmacoeconomics of folic acid supplementation for cardiovascular disease prevention. AB - Epidemiological evidence, linking moderate elevations in homocysteine to cardiovascular events, is compelling but inconclusive. Folic acid and vitamin B12 have been used to reduce homocysteine levels and treat hyperhomocysteinemia. The clinical benefits will be established over the next 5 years as data from randomized trials becomes available. In this paper, two cost-effectiveness analyses examining 'treating all' versus 'screen and treat' are reviewed. Some experts are endorsing screening high-risk patients for elevated homocysteine but most professional societies recommend waiting until more evidence is available from peer-reviewed publications of the results of clinical trials before endorsing population-wide screening or treatment. PMID- 19807362 TI - Economic evaluations of neuraminidase inhibitors to control influenza. AB - Up to 10% of individuals present influenza-like illness each year. Neuraminidase inhibitors significantly reduce the median duration of influenza symptoms by 1.38 days and median time to return to normal activities by 0.9 days in adults. This review presents the economic evaluations of neuraminidase inhibitors in adults. The choice of key parameter estimates in cost-effectiveness and cost-benefit analysis is sensitive to the perspective of analysis: healthcare payer or societal, including productivity gains. This review discusses, among other key parameters, the proportion of influenza-like illness due to the influenza virus (targeted by neuraminidase inhibitors and influenza vaccine), and the measure of health benefits by either quality-adjusted life years gained or willingness-to pay for a day of symptoms averted. Overall, neuraminidase inhibitors are worth their costs and do not challenge annual influenza vaccination but should be seen as a complementary option to reduce the burden of influenza. PMID- 19807364 TI - Impact of the fourth hurdle on the international pharmaceutical industry. AB - Traditionally, pharmaceutical companies were required to provide evidence to demonstrate their product's safety, efficacy and quality for purposes of registration and reimbursement. Increasingly, a fourth hurdle has been added which requires that companies demonstrate the economic value of a product to be reimbursed. This paper provides empirical observations and anecdotes, as well as economic theory to explain the impact of the fourth hurdle on the pharmaceutical industry. Specifically, the paper discusses the impact on drug development, clinical trials, sales, launches, corporate value, human resources and access to new medicines by patients. Using game theory, the potential for suboptimal investment in pharmacoeconomics by the pharmaceutical industry will be explored through a theoretical application of the prisoner's dilemma model. The paper concludes with a hypothetical example for calculating the monetary impact of the fourth hurdle on society when making formulary decisions related to three drugs within a therapeutic class. PMID- 19807365 TI - Measuring quality of life and utilities in esophageal cancer. AB - For many years indicators such as mortality rates, levels of morbidity and cure measured the success of treatments for esophageal cancer. However, it is now recognised that quality of life (QOL) is an important measure of outcome. This is especially so for esophageal cancer, where the therapeutic options include curative treatments or palliative care. A number of measures have been developed but few QOL studies have been conducted in esophageal cancer. A health economics approach to outcome measurement, that seeks to quantify individual preferences (or utilities), offers a number of advantages. However, it is important to recognize that these methods are still under development. Nevertheless, by seeking to measure the strength of individual preferences, utilities provide additional information for decisions regarding which treatments provide the most optimal outcomes. PMID- 19807366 TI - Quality of life outcomes in mental illness: schizophrenia, mood and anxiety disorders. AB - Although health-related quality of life is an amorphous and heterogeneous concept, it has become an important outcome measure in the treatment of psychiatric disorders. This review illustrates the negative effects of distress/clinical factors (psychological distress, anxiety, depressive and negative symptoms) on quality of life of psychiatric patients. Contradictory findings have been reported regarding the relationship of positive symptoms, cognitive deficits and insight with quality of life scores among schizophrenia patients. Increasing evidence suggests that psychosocial or stress process related rather than clinical factors more accurately predict quality of life. The authors suggest that the impact of distress/clinical factors on subjective quality of life of mentally disordered patients is experienced through psychosocial factors. Limitations in current knowledge in this area are identified and suggestions for future research are provided. PMID- 19807367 TI - Costs associated with attention deficit hyperactivity disorder: overview and future projections. AB - The costs associated with attention deficit hyperactivity disorder are considerable. These costs are incurred for the provision of health services, out of-pocket family expenditures, educational services and juvenile justice services. In this review, we present the components of these costs and discuss factors likely to make a significant impact. It is shown that pharmaceuticals and ambulatory care services, in particular outpatient mental healthcare, account for the majority of healthcare costs. Finally, data are presented that suggest attention deficit hyperactivity disorder is associated with increased educational costs, while increased costs associated with juvenile justice may be due to mental health comorbidity. We speculate on future costs for attention deficit hyperactivity disorder given the introduction of new pharmaceutical agents and discuss projected increases in the prevalence of the disorder in under represented populations. PMID- 19807368 TI - Use of HRQOL questionnaires to facilitate patient-physician communication. PMID- 19807370 TI - The START study: when to start to treat with inhaled steroids in asthma? AB - Asthma is an inflammatory disorder with increasing prevalence and significant morbidity worldwide. Inhaled steroids improve asthma symptoms, reduce exacerbations number and severity, and the need for oral steroids. Due to their potent anti-inflammatory effect, inhaled steroids represent the most effective maintenance therapy available and therefore are recommended by all existing asthma management guidelines as the controller medication at all age groups. However, little is known about the effectiveness of the early use of inhaled steroids in mild asthma. The study by Pauwels and colleagues is aimed at assessing the impact of early use of budesonide in patients (both adults and children) newly diagnosed with mild persistent asthma. PMID- 19807371 TI - Bayesian cost-effectiveness analysis based on the persistence with antihypertensive treatment. AB - An observational study was performed to compare five compounds as initial therapy to evaluate the persistence with antihypertensive treatment and drug cost. Over a 1-year follow-up period on the entire population (approximately 360,000 residents) of the Ravenna Local Health Unit. The presence of five major antihypertensive agents between January 1st, and December 31st, 1997 was investigated. There were 4614 patients enrolled. The annual average cost of treatment ranged between euro 44.27 (95% confidence interval, 38.18-52.06) for patients started on atenolol to euro175.65 (150.37-205.10) for those started on losartan. Patients who began losartan showed a higher posterior probability of an effective antihypertensive treatment. On average, these patients turned to produce cost-effective treatment with an average probability of 0.70. PMID- 19807372 TI - Bayesian approach in pharmacoeconomics: relevance to decision-makers. AB - The Bayesian approach is becoming increasingly visible in pharmacoeconomics. This approach has been primarily applied to two decision problems commonly encountered in pharmacoeconomics: adoption and allocation. Acceptability curves generated from Bayesian cost-effectiveness analyses can be interpreted as the probability that the new treatment is cost-effective at a given level of willingness-to-pay. This probabilistic interpretation of study findings provides information that is more relevant and easier to understand for those who make adoption decisions. The Bayesian value of information analysis offers a decision-analytic framework to explore values of additional research and to set research priorities. It is a useful analytical framework for decision-makers who wish to achieve allocation efficacy. PMID- 19807373 TI - Epidemiology, treatment and costs of depression in adults with Type 2 diabetes. AB - Worldwide, diabetes and depression are highly prevalent conditions that are independently-associated with increased healthcare costs, disability, lost productivity, and premature mortality. Approximately 10% of individuals with diabetes have major depression and 30% have both major and minor depression. Individuals with diabetes have two-fold increased odds of having depression compared with individuals without diabetes. Although, there are effective treatments for depression, studies have shown that only 50% of adults with depression are recognized in primary care and that treatment for depression is less than ideal in this setting. The epidemiology, treatment and costs of depression in adults with Type 2 diabetes are reviewed and an expert opinion on the best treatments for depression, and future clinical and research directions in this field of research are offered. PMID- 19807374 TI - Cost-effectiveness of antiemetics use during cancer chemotherapy. AB - Emesis is a major obstacle to cancer chemotherapy. Patients indicate that nausea and vomiting are a substantial concern when receiving chemotherapy. In recent years, newer antiemetics, serotonin antagonists, have been commercialized. Other agents, particularly the neurokin-1 antagonists, are in clinical development and should be available in the near future. Treatment of chemotherapy-induced nausea and vomiting has recently been reviewed by Gralla and Hesketh. The serotonin antagonists currently available are dolasetron (Anzemet), Ben Venue Laboratories, OH, USA), granisetron, ondansetron and tropisetron. They are characterized by an improved efficacy when compared with traditional metoclopramide-based antiemetic regimens. They are particularly effective for the control of acute emesis (within the first 24-h following chemotherapy). Their efficacy for the control of delayed emesis is less impressive and has been shown to be comparable with conventional antiemetics, such as dexamethasone. The serotonin antagonists are also associated with a higher acquisition cost than traditional antiemetics. This has aroused an interest in studying the economic impact of emesis and its treatment. Many economic evaluations have been undertaken since the introduction of the serotonin antagonists. PMID- 19807375 TI - Costs of dyslipidemia. AB - Dyslipidemia has been recognized as an important risk-factor for the development of cardiovascular disease. The current, available therapies of dyslipidemia, their effectiveness, costs, cost-effectiveness and healthcare implications are discussed. At the present time, the lipid-lowering therapies are dominated by statins. Despite a variety of assumptions regarding modeling cardiovascular disease risks and costs, statin therapy is generally cost-effective for secondary prevention and for primary prevention in individuals with additional risk factors. The costs of drug therapy and the absolute risk of developing future cardiovasular events are the dominant factors determining the cost-effectiveness. When developing clinical guidelines, the cost-effectiveness and proportion of the population to be treated must be considered as well as the total population costs of treatment. PMID- 19807376 TI - Impact of transaction costs on healthcare outcomes. AB - This article reviews transaction cost economics to frame a discussion of how inefficiencies in healthcare delivery processes affect clinical outcomes and differentiate between inefficiencies that are tractable from those that are transitional or intractable. Recognizing and quantifying these effects improves the ability of organizations to calculate returns on investment in quality improvement, research and development and related value enhancing, but it is subject to high-risk undertakings. PMID- 19807377 TI - Use of visual analog scales in economic evaluation. AB - Visual analog scales or rating scales are commonly used in economic evaluation to elicit preferences in order to estimate quality-adjusted life years. Values obtained from visual analog scales have been used on their own or via a transformation to map them onto one of the choice-based methods for elicting preferences, namely standard gamble or time trade-off. The arguments against using visual analog scales in economic evaluation directly or indirectly via a transformation are reviewed. It is concluded that it canonly ever provide a second best solution compared with the direct use of a choice-based technique. PMID- 19807378 TI - Fixed and flexible formularies as cost-control mechanisms. AB - The purpose of this review is to consider the prevalent types of fixed and flexible formularies, the general economic principles on which they are based and the evidence for their effectiveness in controlling rising drug expenditures. The principal-agent relationship and economic model underlying the various types of formularies are described. The principal-agent model describes a relationship where there is an asymmetry of information between two parties involved in a particular task. As a result of this asymmetry of information, the party with less information (the principal) allows the party with more information (the agent) to make decisions about that task or activity for them. In the case of formularies and cost-control, the principal is the payer. Depending on the incentives offered by the formulary, the agent can alternately be the prescriber, dispenser or patient. The success of a formulary type to control costs is dependent on two main factors. First, the payer (the principal) must identify the agent for whom it is reasonable to create incentives that incorporate the financial risks associated with use of the drugs. Second, the payer must develop a structure that best aligns the principal and agent objectives. The principal agent framework serves as the vehicle through which the authors examine five major types of formularies (i.e., closed, best available price, reference-based pricing, tiered and open formularies) and their inherent incentives and limitations. The evidence for their effectiveness as cost-control mechanisms is reviewed and the system factors that can affect formulary success will be discussed. Finally, the authors' observations are summarized and interpreted, and suggested implications for future use of formularies in controlling the costs of pharmaceutical use are offered. PMID- 19807379 TI - Behavioral pain assessment and the Face, Legs, Activity, Cry and Consolability instrument. AB - Difficulties assessing pain in individuals who cannot use self-report scales has led to their exclusion from clinical trials and rendered them vulnerable to undertreatment of pain. Although several observational pain scales are available for use in these populations, many lack the characteristics necessary for routine implementation into practice or research. The Face, Legs, Activity, Cry And Consolability pain scale was developed as a simple measure of pain intensity in young children. It has been validated in the postoperative setting in children 2 7 years of age and children aged 4-18 years with varying degrees of cognitive impairment. With minor revisions, the Face, Legs, Activity, Cry and Consolability pain scale may be useful to assess acute pain across populations of children and adults who are unable to self-report their pain. PMID- 19807380 TI - Non-communicating children's pain checklist: better pain assessment for severely disabled children. AB - Advances in the treatment of pain for children with severe disabilities have lagged behind that for other children. This is due, in part, to a lack of valid assessment tools for their pain. The non-communicating children's pain checklists are observational pain tools that were developed specifically for children with severe disabilities who are unable to communicate verbally. The non-communicating children's pain checklist-revised has been validated for use with a wide range of pain types in the home. The non-communicating children's pain checklist postoperative version is used for pain following surgery in the hospital setting. Scores for determining the presence of pain have also been developed. They are appropriate for children with varying degrees of physical, cognitive and communicative impairments. PMID- 19807381 TI - Update on health-related quality of life in patients with gastroesophageal reflux disease. AB - Gastroesophageal reflux disease is highly prevalent in westernized countries. It damages quality of life but its effect is frequently underestimated, often leading to inadequate management of the disease and its symptoms. Health-related quality of life questionnaires are now becoming more widely recognized as an effective means of demonstrating the true impact of gastroesophageal reflux disease, both on individuals and on society at large. Health-related quality of life questionnaires can also help to underscore the importance of effective acid suppression therapy, with proton pump inhibitors, such as omeprazole and esomeprazole, for this common and highly debilitating condition. This review summarizes recent information on the impact of gastroesophageal reflux disease on a range of aspects of health-related quality of life, including patient well being. PMID- 19807382 TI - Measuring health-related quality of life in women on hormone replacement therapy. AB - Hormone replacement therapy has been shown to effectively relieve postmenopausal symptoms. However, its impact on health-related quality of life is not well documented. There is no gold standard instrument available for measuring the effects of hormone replacement therapy on health-related quality of life of postmenopausal women. Furthermore, no systematic comparison of health-related quality of life measures has been undertaken within the field. An overview of the health-related quality of life measures and scales used in publications on hormone replacement therapy are reviewed. In addition, an analysis to compare the performance of two generic health-related quality of life measures, the Nottingham Health Profile and the 15D are conducted. Both measures performed well in postmenopausal women. However, only preference-based measures, such as 15D, can be used in cost-utility analyses. PMID- 19807383 TI - The siren call of new drugs. PMID- 19807385 TI - Intravenous montelukast in asthma exacerbations: could it help? AB - Asthma is a chronic inflammatory disease leading to recurrent episodes of airflow limitation. It has a significant impact both on individuals and society. Pharmacological agents prescribed for asthma management belong to various therapeutic drug classes and are classically administered via the inhalatory route. Recently, leukotriene receptor antagonists have become more widely available and have been shown to produce a variable degree of bronchodilation. Oral montelukast increases the control over the disease in both children and adults and can reduce the dose of inhalatory steroids required in patients with moderate-to-severe and exercise-induced asthma. Recently, intravenous montelukast was shown to improve the airflow in patients with stable asthma more rapidly. However, the role of this formulation in the treatment of acute asthma is unknown. The current study examines the effect of intravenous montelukast in asthma exacerbations. PMID- 19807386 TI - Quality of life in dementia: an ethical and philosophical perspective. AB - Quality of life is crucial when thinking of interventions in dementia. Over the last 10 years huge efforts have been made to develop methods of measuring quality of life. The development of such measures faces significant conceptual and methodological difficulties. Despite the advances made in this field, conceptual questions remain. Indeed, the conceptual limits to the measurement of quality of life lead the author to conclude that quality of life, as such, is not measurable. Nevertheless, specific aspects of quality of life can be measured but as the constructs broaden, the need for a phenomenological approach increases. PMID- 19807387 TI - The cost-effectiveness of short-term dynamic psychotherapy. AB - Short-term dynamic psychotherapy is a group of well-researched brief treatments with over 50 published controlled trials testing its effectiveness against a range of treatment and nontreatment controls. Studied samples are often high users of medical services, hospital services, mental health services and disability insurance. If short-term dynamic psychotherapy is effective, it should translate into demonstrable reductions in medical and social system costs. This review examines whether or not short-term dynamic psychotherapy is a cost effective treatment. PMID- 19807388 TI - Risk-adjusted capitation payment systems for health insurance plans in a competitive market. AB - Adequate risk-adjustment is critical to the success of market-oriented healthcare reforms in many countries. Regulated competition among insurers and providers is often a crucial element in these reforms. Consumers may then choose among competing health insurance plans, which are largely financed through premium replacing risk-adjusted capitation payments. Too crude risk-adjustment, which is common in 2003, provides health insurance plans with incentives for risk selection, which may threaten access to (good quality) healthcare, efficiency and consumer satisfaction. In this review the background and rationale for the implementation of risk-adjusted capitation payments is described. The authors are optimistic about the possibilities for improving the present, crude risk-adjusted capitation payment systems in order to reduce risk selection. PMID- 19807389 TI - Measuring patients' assessments of primary care quality: the use of self-report questionnaires. AB - The views of patients are seen as an increasingly important complement to other measures of quality of care, such as clinical indicators. This review summarizes previous research and current issues relating to the use of patient assessments of primary care quality. Patient assessments can be used to measure a number of different domains of primary care quality. Significant advances have been made in terms of the production of comprehensive, reliable and valid patient assessments, which can be used in both research and quality improvement activities. However, the effectiveness of the use of patient assessments as a technology for quality improvement remains unclear. PMID- 19807390 TI - Assessing the equality of means of healthcare costs. AB - The main analytic problems for assessing the equality of means of healthcare costs are the skewed and heteroscedastic nature of the distribution of healthcare costs and the occurrence of persons with zero cost values. Without taking these special features into consideration, any statistical method may lead to the wrong conclusion. The available analytic methods that are appropriate for handling these features of healthcare costs are reviewed. PMID- 19807391 TI - Caution in the use of the apolipoprotein E e4 allele as a predictor of healthcare costs. AB - Healthcare payers of all types are interested in accurately predicting future costs. The e4 allele of the gene coding for apolipoprotein E on chromosome 19 (e4 allele) is a potentially attractive genetic marker. This could be used to set insurance premiums or determine whether an insurance policy is actually underwritten because it is a susceptibility marker for Alzheimer's disease, heart disease and stroke. There is some evidence that the apolipoprotein E e4 allele is associated with healthcare costs. However, caution is warranted given the limited number of studies that associate the e4 allele and cost, and concerns regarding selection bias in studies that show an association between the e4 allele and occurrence of costly diseases. Until more is known, the e4 allele should not be used to approve or deny issuance of insurance or to set premium rates for either acute or long-term care insurance. PMID- 19807392 TI - Pharmacoeconomic considerations of tropisetron for prophylaxis and treatment of CINV and PONV. AB - Postoperative nausea and vomiting has been described as the big little problem in anesthesia, with the overall incidence after anesthesia using volatile anesthetics remaining between 20 and 30%. In patients who receive ambulatory surgery, postoperative nausea and vomiting may lead to delayed discharge or unplanned overnight admission to hospital which is counterproductive to the primary goal of saving costs in healthcare by undergoing day-surgery. The same economic factors apply to patients who experience nausea and vomiting after chemotherapy. Drugs today are judged by their ability to cure a condition with as few adverse reactions as possible. There remains the question of whether a certain medication meets expectations from a pharmacoeconomic point of view. 5 HT3 antagonists are a comparatively new class of drugs that seem to perform satisfactory in the treatment of different forms of nausea and vomiting with few adverse reactions. However, these drugs are comparatively expensive. Studies on the efficacy and pharmacoeconomic comparisons have been conducted. Tropisetron is one of the newest 5-HT3 antagonists. While its efficacy has been shown in several studies, there are few studies on its pharmacoeconomic benefits. There are several antiemetic measures, such as total intravenous anesthesia, using antiemetic prophylaxis or omitting the nitrous oxide with proven effectiveness. Most are roughly equivalent but differ with respect to costs and side effects Routine antiemetic prophylaxis is not indicated due to economic and medical reasons (potential side effects of antiemetics). Patients at high risk can be identified using validated risk scores and should receive antiemetic drugs. PMID- 19807393 TI - Management of Crohn's disease from efficacy, quality of life and health economic perspectives. AB - The expectations of the physician and patient in the modern management of Crohn's disease have moved on from alleviation of symptoms whenever relapses occur, to sustained maintenance of remission and mucosal healing. The majority of treatment costs are as a consequence of hospitalizations and surgery. These heightened therapeutic goals are leading to expectations that management costs will decrease. Health-related quality of life can be measured by validated disease specific tools in Crohn's disease, and such measurements are now forming common outcome measures in clinical trials of new therapies in Crohn's disease. This review gives an overview of patient-focused management of Crohn's disease and the rapid evolution in therapy, with pharmacoeconomic analysis where available. PMID- 19807394 TI - Update on the use of patient reported functional status and HRQOL measures in low back pain. AB - Disease-specific functional status and generic health-related quality of life instruments used in low back pain studies are reviewed. A variety of instruments are available and many have been demonstrated to be valid and reliable. The Roland-Morris Disability Index and Oswestry Disability Index are the most commonly used low back pain-specific measures, while the Short Form-36 is the most commonly used generic quality of life tool. The Short Form-36 is valid and reliable in back pain but not necessarily intended to replace disease-specific measures. Studies have been inconsistent in their efforts to demonstrate a relationship between self-reported functioning and task performance. PMID- 19807395 TI - Health-related quality of life with chronic leg ulceration. AB - Chronic leg ulceration is a major cause of morbidity in the elderly. The existing evidence on the evaluation of health-related quality of life for patients suffering from this condition is reviewed. Research evidence was categorized into qualitative research; development and validation of disease-specific tools; cross sectional quantitative methods; and longitudinal and outcomes studies. Major themes associated with leg ulceration include ulcer pain, poor sleep quality, mobility deficit and the resulting consequences on the patients' social lives. Patients frequently adapt their lifestyles to avoid situations that may lead to further ulceration. While elderly women experience the poorest quality of life, it appears that leg ulceration has the greatest impact on young men. Greatest improvements over time are achieved for patients whose ulcers heal completely. Leg ulceration has a major impact on patients' health-related quality of life. Studies must consider this as an important outcome, particularly when complete healing is unlikely. PMID- 19807396 TI - Quality of life assessment in osteoarthritis. AB - Optimal prevention and therapy management depends upon the quality of health assessment in the field of increasingly frequent degenerative health disorders. Quality of life has increased in importance as a result of World Health Organizations' new concept of health and disease. This review addresses strategies and measurement in the quality of life assessment in osteoarthritis over the past 15 years. The authors produce an overview of valid quality of life instruments and their application on patients with hip or knee osteoarthritis, focused on the responsiveness of the tools and on the effect of different interventions. Continuous improvement of therapy and result quality must remain adjusted to the patient and must involve cost carriers in each individual situation to attain the best quality of life under the given socioeconomic conditions. PMID- 19807397 TI - Quality of life assessment for chronic stable angina. AB - In this article, the reasons for quality of life assessment in people with chronic stable angina are reviewed. A table of instruments most commonly used to assess quality of life for people with ischemic coronary disease is provided. Studies that examine quality of life for people with several types of ischemic coronary disease, including chronic stable angina and myocardial infarction, are discussed. Studies that examine instruments used to evaluate quality of life for people undergoing revascularization for ischemia and those enrolled in cardiac rehabilitation are also addressed. Finally, studies that focus on quality of life in women with coronary disease are commented on. PMID- 19807398 TI - Quality of life issues associated with antipsychotic-induced weight gain. AB - For patients with mental illness, the purpose of antipsychotic medication is to improve their quality of life, and for many, quality of life improves dramatically with treatment. However, weight gain in the range of 7 to 10% is commonly associated with the use of antipsychotic medications and can have a negative impact on patient quality of life. Being overweight or obese can lead to serious medical conditions, including insulin resistance, diabetes mellitus, hypertension, dyslipidemia, arthritis and coronary heart disease. The physical and psychological consequences of weight gain are an added burden for patients with schizophrenia or other mental disorders. Successful strategies to prevent and manage weight gain include diet and exercise interventions and, in some cases, switching to a different antipsychotic agent. PMID- 19807399 TI - Quality of life and rhinitis: nothing to be sneezed at! PMID- 19807401 TI - Decision-makers' use of pharmacoeconomics: what does the research tell us? AB - Governments, organizations and industry routinely make decisions regarding the value of pharmaceuticals. The perspectives, techniques, decision constraints and available information differ across these decision-makers and sometimes within categories. Although pharmacoeconomics is consistent with the international trend toward evidence-based decisions in medicine and could benefit all of these decision-makers, the potential impact of these studies has been largely unrealized. Why is it that a field to which so much research funding has been devoted, has produced so little of clear use to major decision-makers? Would inflexible organizational barriers surrounding narrowly defined performance incentives and line item budget accountability limit the usefulness of even impeccably performed pharmacoeconomic studies? Or, are methodological and logistical considerations impeding the usefulness of pharmacoeconomics? PMID- 19807402 TI - Cost-effectiveness of screening for genital Chlamydia trachomatis. AB - Chlamydia trachomatis is the most common, curable, sexually transmitted disease in many Western countries, leading to severe sequelae, such as infertility and ectopic pregnancy. As most chlamydial infections are asymptomatic, screening programs seem to be an attractive public health measure. MEDLINE, EMBASE, the Health Economic Evaluation Database and the National Health Service Economic Evaluation Database were searched for economic evaluations of Chlamydia trachomatis screening programs. Key factors influencing the cost-effectiveness are identified and assessed, such as screening strategy, test system and treatment regimen. Standard and new methodological approaches for assessing the cost-effectiveness are presented and future developments in the field are predicted. The most cost-effective screening approaches are compared with the recommendations of international guidelines. PMID- 19807403 TI - The cost-effectiveness of screening for colorectal cancer. AB - Screening policies for colorectal cancer are costly, as they are to be applied to a large potential population. Cost-effectiveness analysis of potential screening policies is therefore warranted and depends on local circumstances and healthcare systems. Most studies have used modeling approaches, with a few exceptions on the use of fecal occult blood tests. Current conclusions of economic studies tend to favor either double barium contrast enema or sigmoidoscopy as a mass screening tool, although colonoscopy might prove cost-effective in some circumstances. Further research is needed to assess the cost-benefit of mixed strategies in large populations. PMID- 19807404 TI - Assessment of the economic value of epoetin use in anemic cancer patients. AB - Anemia is a common occurrence in cancer patients, as a consequence of the disease process and treatment. Until recently, the only treatment available for anemia was red blood cell transfusions, but this was withheld until the development of severe anemia because of the risks associated with transfusion. Since the mid 1990s, recombinant human erythropoietin (rHuEPO) has been available as an alternative treatment option, which could be used to treat mild-to-moderate anemia and to prevent severe anemia. Many clinicians currently believe that anemia should be treated vigorously, because this is thought to improve patient quality of life, partly because of new research indicating that anemia may be an independent risk-factor for survival. Possible mechanisms behind such an association are not yet completely elucidated, but some clinicians argue that rHuEPO is an ideal drug for the treatment of mild and moderate anemia, as it is perceived to be without serious negative side effects. However, rHuEPO is an expensive drug and the evidence on how to use it optimally, is still highly incomplete. PMID- 19807405 TI - Economic burden of asthma: implications for outcomes and cost-effectiveness analyses. AB - Asthma is a disease of chronic airway inflammation. It is of importance to clinicians and health systems because the hospitalization and death rate due to asthma have increased since 1980. Cost of illness studies have estimated that the total cost of asthma (direct and indirect costs) exceed USD 10 billion annually, in the USA. Since 1985, the proportion of asthma costs in hospitals have decreased and the proportion of costs due to asthma medications have increased. However, approximately half of direct medical costs of asthma are due to hospitalizations. The mean direct cost of asthma per year per patient has been estimated to be approximately USD 1,100. As the implementation of national and international guidelines continues, future costs for asthma will likely come from the treatment and management of the disease. Adequate assessments of treatment and cost-effectiveness analysis are important. Recommendations promoting the use of cost-effective anti-inflammatory medications are crucial to efficiently managing asthma. PMID- 19807406 TI - Syncope and falls in older people: defining the size of the problem. AB - The proportion of the population over the age of 65 is increasing. Reducing diseases that are common in this age group and that are associated with morbidity and mortality is a priority, if healthcare budgets are to be used cost effectively. Syncope and falls are common in older people and are a major cause of healthcare expenditure. Clinical interventions are available, but the cost effectiveness of those interventions is unclear. PMID- 19807407 TI - New aspects in health economic studies of prevention and treatment of osteoporosis. AB - This review updates the current knowledge with regard to important new clinical data in the field of osteoporosis interventions and discusses the implications for future health economics studies in this field. PMID- 19807408 TI - Contemporary measures for the assessment of health-related quality of life in prostate cancer. AB - Over the past decade, increasing attention has been paid to the proper assessment and evaluation of health-related quality of life in men with prostate cancer. A major motivation for this work is the lack of strong evidence that survival end points differ among the various therapies in the setting of documented differences in morbidities. This review summarizes the current literature regarding the evaluation of health-related quality of life for prostate cancer. The use of validated measures and other issues relevant for prostate cancer specific assessments will be discussed. PMID- 19807409 TI - Measuring the value of program outcomes: a review of multiattribute utility measures. AB - Selection of a multiattribute utility instrument for economic evaluation is complex due to competition between developers and inflated claims for instrument properties. This review presents multiattribute utility theoretical requirements and assesses leading multiattribute utility instruments against these. Instruments reviewed are: the QWB, HUI3, 15D, EQ5D, AQoL and SF6D. The key finding is that no current instrument satisfies all the requirements for multiattribute utility measurement. We recommend that users should choose instruments most relevant to their circumstances and that studies should include two instruments. Rigorous sensitivity analyses should be conducted and both results reported. Subject to these caveats, preference should be given to instruments best meeting multiattribute utility theoretical requirements, viz., the AQoL or HUI3. However, we recognise that other instruments may perform as well, or even better, under certain circumstances. PMID- 19807410 TI - Some reflections on cost-effectiveness analysis and budget allocation in medicine. PMID- 19807412 TI - Determinants of costs in blood services: blood transfusion from an economic perspective. AB - Over the past decade, technology-driven inflation of blood transfusion costs has largely exceeded the medical component of the consumer price index. This tendency may speed up in the near future, despite poor cost-effectiveness projections and the low capacity of new technologies to produce clinically appreciable quality improvements or an increased productivity. Although labor is the costlier input, there has been little research and development aimed at substituting capital for labor in blood services. Substantial advances in efficiency can come only from a whole process re-engineering and adoption of a societal perspective in decision making. There are, however, many barriers to such changes. Notably, the monopolistic nature of blood services, the perverse incentives derived from their current structure, with an increasing gap between the interests of producers and users, pressures by industry, cultural perceptions about blood and the political and judicial consequences of the transfusion AIDS epidemic. PMID- 19807413 TI - Economic evaluation of noncontact normothermic wound therapy for treatment of pressure ulcers. AB - New adjunctive treatments for pressure ulcers have become available to complement standard care. The economic benefits of new advanced wound care treatments like noncontact normothermic wound therapy are related to: the costs of adequately providing standard care treatment, the baseline probability of healing a pressure ulcer to closure with standard care, the relative improvement in healing rates with the advanced wound care treatment and the acquisition cost of the advanced treatment. Healing data from preliminary clinical trials suggest that pressure ulcer healing in long-term care patients is accelerated two-fold with noncontact normothermic wound therapy. At this healing rate, noncontact normothermic wound therapy for stage III and IV pressure ulcer is an economically attractive intervention. Additional well-controlled clinical trials are necessary. PMID- 19807414 TI - Cost-effectiveness of alternative strategies to prevent trachomatous blindness. AB - The Alliance for the Global Elimination of Blinding Trachoma was formed in the mid 1990s. The Alliance of country representatives and experts on trachoma suggested a four-pronged approach to eliminate incident trachomatous blindness: surgery for trichiasis, antibiotics to treat the disease, facial cleanliness and environmental improvements to limit disease transmission. The efficacy and effectiveness of the components of this approach have been evaluated, but the strategy as a whole has not been compared with different combinations of its components. The relevant cost-outcome and pharmacoeconomics literature is limited. This article reviews the cost-effectiveness of the approach's components. Furthermore, the article mentions ongoing research that will address the cost-effectiveness of the entire strategy for eliminating a major cause of preventable blindness in the developing world. PMID- 19807415 TI - Cost-effectiveness of replacing NSAIDs with coxibs: diclofenac and celecoxib in rheumatoid arthritis. AB - Cyclooxygenase-2 inhibitors exhibit the same effectiveness on rheumatic symptoms as nonsteroidal anti-inflammatory drugs, have fewer gastrointestinal side effects, but a higher price. Based on cost and healthcare utilization in Norway, this paper explores what factors are most important for the cost-effectiveness of cyclooxygenase-2 inhibitors and points to patient groups that may be most relevant for prescribing coxibs instead of nonsteroidal anti-inflammatory drugs. PMID- 19807416 TI - Societal implications of the pharmacoeconomics of alpha1-antitrypsin deficiency. AB - The use of intermittent augmentation therapy for patients with alpha(1) antitrypsin deficiency assists in slowing disease progression and may, as a result, reduce the likelihood of hospitalization and significant clinical events. There are significant cost and access issues related to current therapy management, since currently marked augmentation therapy is expensive and there have been periodic shortages. The high cost, access and reimbursement concerns have prompted questions about the potential pharmacoeconomic advantages of future treatments. In this review, we provide an overview of published research findings concerning the effectiveness of current therapy, reimbursement and cost issues and potential future advances in treatment for alpha(1)-antitrypsin deficiency. PMID- 19807417 TI - Pharmacoeconomics in oncology. AB - Healthcare costs in the USA continue to rise faster than the consumer price index. Nothing demonstrates this more vividly than the double-digit increases posted for the cost of the drug treatment of the oncology patient. A factor that will compound this cost is the expansion in the oncology patient population that will occur as the population ages. Pharmacoeconomics is a discipline that evaluates the relationship between clinical, economic and humanistic outcomes to determine the products and services that maximize the value for each dollar spent. Research in this area is evolving to meet the needs of the individual patient and decision-makers within a payer group, healthcare system, or society. Healthcare interests in countries in Europe, Canada and Australia have already adopted analytical tools and incorporated them into guidelines for drug use. The USA is also moving in this direction now that the Food and Drug Administration is considering requiring studies in pharmacoeconomics in addition to the standard studies of the safety and efficacy of drugs. The importance of this approach to oncology will be seen as policy-makers apply research findings to practice decisions. PMID- 19807418 TI - Cultural issues in assessing quality of life in cancer clinical trials. AB - Over the past decade, there has been significant progress made into the field of quality of life in cancer clinical trials. Many cancer clinical trials now include quality of life as a secondary end-point, but only a very small number of trials include quality of life as a primary end-point. This article reviews and discusses issues that arise when developing a quality of life instrument for use in international cancer clinical trials. We describe problems that arise when trying to establish item equivalence across cultures that the instrument development team should be aware of. We also outline approaches in the literature to establish item equivalence across different cultural groups both in a linguistic and conceptual manner. Finally, statistical and psychometric approaches to assess cultural differences and cultural equivalency that have appeared recently in the literature are described and reviewed. PMID- 19807419 TI - Integrated health outcomes research strategies in drug or medical device development, pre- and postmarketing: time for change. AB - The implementation of health outcomes research as a healthcare decision-making tool has expanded rapidly in the last decade. Drugs and medical devices are increasingly being required to demonstrate not only their efficacy and safety characteristics, but also their performance in at least three core dimensions of health outcomes research: clinical effectiveness, patient-reported outcomes and economic outcomes. However, the current integration of health outcomes research lacks coordination and communication and as a result, money and time is being spent on the generation of health outcomes research data which can be both insufficient and fail to satisfy the information demands of all the relevant stakeholders. In response to this, a new paradigm is evolving which involves the implementation of health outcomes research strategies that encompass the development, pre- and postmarketing stages of a drug or medical device. PMID- 19807420 TI - Future of healthcare reform in the USA: lessons from abroad. AB - Healthcare reform in any nation is an evolving process. Brought about by demographic, technological, social, cultural, economic and political factors, all healthcare systems are continually confronting issues related to cost, access and quality. This paper examines other countries' approaches to healthcare in the ongoing efforts at healthcare reform in the USA. While recognizing the uniqueness of the healthcare system in each nation, it appears there are valuable lessons from other nations to be considered by policy makers in the USA as healthcare reform continues to evolve. This paper synthesizes several lessons for the USA that may be applicable by looking beyond its borders. In so doing, it reveals differences that may be insightful in considering future healthcare paradigms influencing healthcare reform efforts in the USA. PMID- 19807422 TI - Cost-utility analysis: the nirvana to health economists and an enigma to decision makers? PMID- 19807421 TI - Closing the credibility gap in patient-reported outcomes. PMID- 19807423 TI - Confusion about European Organisation for Research and Treatment of Cancer Quality of Life Prostate module (EORTC QLQ-PR25): a response. PMID- 19807425 TI - Pharmacoeconomics of antipsychotic medications. AB - Antipsychotic medications play a central role in the treatment of schizophrenia and other psychotic disorders. In the last decade, second-generation antipsychotic agents have been introduced that are more expensive on a cost per daily dose basis than older agents. We review empirical data showing trends in prescribing patterns of first- and second-generation antipsychotic agents in the USA and the cost implications of these trends. We show how drug acquisition costs fit into the matrix of overall treatment costs, including pharmacy costs, in and out patient treatment and laboratory costs. We examine specific incentives that influence payers in these various domains and consider how incentives and policies might be constructed so that they serve to encourage effective treatment. PMID- 19807426 TI - Economics of family planning and birth control. AB - The 50th anniversary of the Pill proves a useful setting against which to consider developments in contraception and birth control. However, differentials between the demands placed on health services and the resources available to meet such needs continue to be major headaches for those involved in policy making and the planning, provision and delivery of services. This review considers the cost effectiveness of contraception and in particular, long-term reversible devices, which are effective in preventing unintended pregnancies and avoid the need for compliance. The paper discusses the cost-effectiveness of contraceptives and the economic benefits generated when unintended pregnancies are avoided. It also highlights the major inequities that exist in access to contraceptives on a global basis and highlights the need to increase provision in developing countries. PMID- 19807427 TI - Microcosting versus DRGs in the provision of cost estimates for use in pharmacoeconomic evaluation. AB - Investigating the pharmacoeconomic impact of any diagnostic or therapeutic intervention in the Irish healthcare setting is currently compromised by the lack of detailed cost data. Consequently, we conducted a number of microcosting studies in the areas of acute myocardial infarction, cardiac failure and HIV, from the hospital perspective. The results of these microcosting studies were compared with the costing estimates assigned to hospital admissions, based on the diagnosis-related group system. Differences ranged from 9 to 66%. It was concluded that the diagnosis related group system is a useful estimate of costs for patient admissions in the absence of detailed cost of illness data. However, supplementary costing studies should be performed for certain therapeutic areas, particularly those where investigation and/or treatment costs are high. PMID- 19807428 TI - Efficiency-based pharmacotherapy: a new approach for decision-making in medical practice. AB - For many years, efficacy and safety have been key parameters for decision-making in healthcare policy. However, the current situation is rather different as resources are scarce, so it is necessary to take efficiency into account when allocating limited resources. Efficiency-based pharmacotherapy is a new discipline that combines the principles of cost-effectiveness analysis with the criteria of evidence-based medicine to obtain the efficiency of therapeutic options to treat important diseases. A league table for each important pathology will be created by ranking all available drugs according to their incremental cost-effectiveness ratio. In this way, clinicians and other decision-makers would be able to advice the use and implantation of drug therapy options with better value for money in routine medical practice. PMID- 19807429 TI - Pharmaceutical pricing in the developing world: issues of access to medicines. AB - Events during 2001 raised greater awareness in the global community to the problems facing less developed countries in gaining access to essential medicines because they are not affordable. The difference between the price of medicines and income in many developing countries is significant and consequently, price stands as an important barrier to access. Steps taken this year to clarify the flexibility clauses of the Trade-Related Aspects of Intellectual Property Rights Agreement - international discussions on the feasibility of applying a differential pricing mechanism and the Global Fund to Fight AIDS, Tuberculosis and Malaria - have been important advances in the process that seeks to ensure access to affordably priced medicines. This paper reviews recent developments and proposals concerning issues of drug pricing in developing countries and discusses what still needs to be done in order to improve access. PMID- 19807430 TI - Toward new legislation on drugs in Europe. AB - Everybody acknowledges the merits of the European Medicines Evaluation Agency and the progress the European system has permitted in evaluating and approving medicinal products. Nevertheless, the system established in 1995 is ripe for updating and improvement in the light of experience. Some limitations of the procedures suggested by the european Community regulations seem to hamper achievement of the real aims. The scientific community needs to identify the problems and propose and debate solutions with the politicians concerned, so that the european regulatory authority can fulfil its commitment to the defense of public health. The European Commission has made its proposal for improving the system. Here are our first comments. PMID- 19807431 TI - Quality of life in breast cancer patients: measurement issues in cancer clinical trials. AB - Quality of life is becoming a significant issue to measure in breast cancer patients. Increasingly, in breast cancer clinical trials we see quality of life as a secondary end-point and we are understanding more about the needs of these women. However, as yet, there is no gold standard regarding which tool is the most appropriate to use. This article reviews quality of life measures that have been used in cancer clinical trials with breast cancer patients and evaluates their reliability, validity and cultural relevance. We propose suggestions for selecting future measures for use in a breast cancer population of patients. PMID- 19807433 TI - Use of economic models in formulary decisions. PMID- 19807432 TI - Fatigue assessment in cancer clinical trials. AB - Studies repeatedly show that fatigue correlates directly with overall quality of life, greater fatigue leading to poorer outcomes. A considerable number of studies reported in the last decade stress the complex problem faced by cancer patients experiencing fatigue, when undergoing treatments and/or following a course of treatment. Fatigue can continue for many years after cure and for those patients with an active or progressive disease, fatigue can lead to a loss of overall quality of life that is limiting in the extreme. While researchers continue to develop new treatments and approaches that can have benefits in terms of survival, it is important to understand how fatigue affects cancer patients and how it can be addressed in cancer clinical trials. PMID- 19807435 TI - Method without madness. PMID- 19807436 TI - Change and stability in the Canadian healthcare system. PMID- 19807437 TI - Indirect medical costs in the first 3 years of rheumatoid arthritis: comparison of current methodological approaches. AB - Indirect cost of chronic diseases have received increasing attention since recent diagnostic and therapeutic improvements have markedly raised costs of care. Relevant cost savings can be expected through the reduction of indirect costs. However, the assessment methods of indirect cost components still differ widely, leading to heterogeneous data that do not allow for a direct comparison. This prospective study of gainfully employed patients suffering from rheumatoid arthritis aims to compare indirect costs assessed by different current methodological approaches (human capital and friction cost method valued by either population or individual data), and compare the methods employed in order to provide a proposal for the standardization of indirect cost assessment. The finding of the present investigation underline the importance to define standardized assessment methods for a comparison of the results of cost evaluations comprising indirect costs. A recommendation for the assessment of productivity losses and their valuation is given. In a given specific economic investigation, the optimal assessment method might differ from these general statements. These recommendations are proposed to give an orientation in a field of heterogeneous strategies in order to improve comparability of indirect cost assessment. PMID- 19807438 TI - Discrete choice experiments to measure consumer preferences for health and healthcare. AB - To investigate the impact of health policies on individual well-being, estimate the value to society of new interventions or policies, or predict demand for healthcare, we need information about individuals' preferences. Economists usually use market-based data to analyze preferences, but such data are limited in the healthcare context. Discrete choice experiments are a potentially valuable tool for elicitation and analysis of preferences and thus, for economic analysis of health and health programs. This paper reviews the use of discrete choice experiments to measure consumers' preferences for health and healthcare. The paper provides an overview of the approach and discusses issues that arise when using discrete choice experiments to assess individuals' preferences for health and healthcare. PMID- 19807439 TI - Economic impact of poor compliance with pharmaceuticals. AB - Poor compliance with prescribed drug regimens is a widespread phenomenon which results in decreased efficacy and is often associated with increased medical expenditures due to treatment failure and/or adverse events. This review examines the issues relating to the measurement of compliance and the clinical and economic consequences of noncompliant drug taking behavior. The results of a literature review, identifying pharmacoeconomic evaluations which measured compliance, are presented. It concludes by highlighting the importance of accounting for poor compliance in pharmacoeconomic evaluations and suggests ways in which compliance may be incorporated in economic models. PMID- 19807440 TI - Update on the use of osteoarthritis-specific outcome measures. AB - Osteoarthritis is the single most important cause of disability and limitation of activity of elderly people in the UK. Although joint replacement is now commonplace for hips and increasingly so for knees, pharmacological management continues to be important. In this context, it is necessary to identify valid and acceptable outcome measures to enable progress in treating osteoarthritis to be evaluated. Such measures should benefit not only clinicians managing osteoarthritis and purchasers of healthcare for this condition, but, ultimately also patients through improved forms of treatment. This paper briefly reviews and provides an update on the use of osteoarthritis-specific health and functional status and generic health status outcome measures in health services research and technology assessment. Other equally important clinical outcome measures, such as biochemical markers, radiographic assessment and joint imaging, are not discussed. PMID- 19807441 TI - Impact of antipsychotic agents and their side effects on the quality of life in schizophrenia. AB - Schizophrenia is the most chronic and disabling of mental illnesses and it imposes a disproportionately large economic burden on society. In addition to symptom reduction and cost-effectiveness, quality of life is becoming a critical outcome measure for the efficacy of antipsychotic agents in the treatment of schizophrenia patients. This review concentrates on the influences of antipsychotic agents and their side effects on quality of life of schizophrenia patients. Of the 182 papers elicited in a comprehensive Medline search from 1996 2002, we found only 21 significant papers (11.5%) reporting findings concerning the effect of antipsychotic agents on quality of life of schizophrenia patients. Very few studies directly examined the impact of side effects on quality of life ratings of schizophrenia patients. To date, there is no clear evidence that in the long-term, atypical antipsychotics are more effective or are better tolerated than typical antipsychotics. This review suggests that side effects of antipsychotic agents influence subjective quality of life of schizophrenia patients significantly less than clinical and psychosocial factors. The patient's subjective response to side effects of medication is more predictive of quality of life than the number of those effects. We discuss both patients' and clinicians' perceptions of side effects and quality of life, which should be considered in the evaluation of the efficacy of antipsychotic agents. PMID- 19807442 TI - Health-related quality of life issues in urinary urge incontinence. AB - Urinary incontinence is a highly prevalent condition which affects the physical, social, psychological and economic well-being of individuals and their families. The assessment of health-related quality of life has been increasingly applied to urinary incontinence. Of the three major types of urinary incontinence (stress, urge or mixed), urge and mixed urinary incontinence have been proven to have the worst effect on health-related quality of life. This paper reviews the health related quality of life questionnaires designed specifically for urge urinary incontinence patients, describing their main characteristics and limitations. Four additional health-related quality of life questionnaires for urinary incontinence patients are also described, either because of their widespread use or because of their proven utility in the evaluation of urge urinary incontinence. PMID- 19807443 TI - Meta-analysis of rare and adverse event data. AB - Potentially beneficial drugs may have side effects. Such adverse events may be serious but rare, making their effect difficult to quantify, or even their identification problematic within randomized controlled trials. Meta-analysis provides a means of combining such outcome data across a series of studies and provides a more powerful analysis than is possible from a single study. However, meta-analysis of adverse event data presents some unique methodological challenges. Procedural issues pertaining to study selection and the inclusion of both randomized and observational data are discussed. The statistical problem of combining rare outcomes is also considered at length. Model choice, continuity corrections, exact statistics, Bayesian methods and sensitivity analysis are all covered. The application of results from adverse event meta-analyses to clinical and economic decision models is also considered. A number of examples are presented to illustrate the key points. PMID- 19807444 TI - Evaluation criteria for pharmacoeconomic and health economic Internet resources. AB - The purpose of this paper is to present criteria for evaluating Internet sites and supply a selective sample of sites that may be useful in improving the transfer of information and diffusion of innovation in pharmacoeconomics and health economics. The nine criteria deal with the following: 1) Expertise - Qualified individuals should generate the information. 2) Confidentiality - All patient data should be confidential and protected. 3) Referencing - Information should be supported with proper citations. 4) Justification - Rigorous evidence should support the claims relating to benefits, risks and costs of any intervention or service. 5) Authorship - provided in the clearest manner with contact addresses. 6) Sponsorship - Support for sites should be clearly identified. 7) Advertising & Editorial Policy - Advertising should be distinguishable from other informational content. 8) Study Evaluation - should comply with the standard economic evaluation checklists 9) Generalizability - Sites should define their target countries and audiences and discuss potential generalizability. PMID- 19807446 TI - Cost-of-illness analyses for policy making: a cautionary tale of use and misuse. PMID- 19807445 TI - Pharmacy benefit design options available to employers. AB - I explore all the pharmacy benefit options currently available to employers. First, I discuss the design of the benefit structure, which can involve up to five tiers of drug options and three tiers of pharmacy options. Next, I discuss how the employer can overlay a cost-sharing structure on top of that benefit structure, in order to provide each employee with the right balance of choice and incentives. The cost-sharing structure can entail up to seven main types of cost sharing: copayments, coinsurance, incentive pricing, benefit caps, quantity caps, deductibles and defined contributions. PMID- 19807448 TI - Quality of life following lung cancer surgery: what about before? AB - Lung cancer is the major malignancy worldwide with cigarette smoking being the main risk-factor. In lung cancer patients, quality of life is considered the strongest prognostic factor for survival irrespective of initial performance status, weight loss, stage of disease, number of metastatic sites, and type of treatment. The study evaluates the health-related quality of life of patients after surgical intervention using a generic questionnaire, the Medical Outcome Study Questionnaire Short Form, and compares this outcome with a normal healthy population, lung cancer patients, and coronary artery bypass grafting patients. Compared with healthy controls and coronary bypass patients, lung cancer patients had significantly higher impaired physical functioning scores, comparable with mean scores for anxiety and depression. However, health-related quality of life in lung cancer remains an issue which needs further assessment. PMID- 19807449 TI - Neck Pain and Disability Scale: a critical evaluation. AB - This report provides a review of the current procedures for measuring cervical pain and dysfunction and discusses some of the methodological issues involved in the measurement of chronic pain. In particular, the Neck Pain and Disability Scale is compared with other measures of pain and disability. Suggestions are made for further documentation of clinical and research usefulness. PMID- 19807450 TI - Global cost modeling analysis of HIV-1 and HCV viral load assays. AB - This review addresses hidden costs associated with the Bayer VERSANT assay, Roche AMPLICOR MONITOR test and COBAS AMPLICOR MONITOR test and how these influence the final per reportable cost to a testing laboratory in resource-rich and -poor countries. An in-depth evaluation and recommendation of the most cost-effective approach for these tests is presented. The analyses demonstrate the need for manufacturers to consider labor and supply costs when marketing a kit in resource poor countries, noting that marketing strategies need to change. In the absence of any proven monitoring alternative, emphasis is placed on increasing market share to promote significant reduction in kit prices to suit the demands of markets in resource-poor countries. Finally, recommendations are made to improve the overall cost structure of viral load testing. This review is intended as a tool to optimize assay usage in attaining the lowest performance costs by assay and is not to endorse any test, as will become apparent. PMID- 19807451 TI - Modeling cost-effectiveness of HIV prevention programs. AB - There have been important advances in the literature regarding modeling the cost effectiveness of HIV prevention programs. Frequently, costs and/or effectiveness are modeled with a combination of parameters since they cannot be measured directly. In most HIV prevention interventions, it is not possible to directly measure effectiveness due to changes in HIV incidence. Instead, effectiveness in preventing HIV is usually assessed using mathematical models that translate measurements of behavior change into estimates of the number of HIV infections averted. This measure of HIV prevention effectiveness is dependent upon the risk levels in the communities (e.g., prevalence of HIV, sexual or drug-injection behaviors), and on the behavioral effectiveness of the intervention in changing participants' risk behaviors. The number of HIV cases averted can be calculated using probabilistic models of HIV transmission, which compare the sexual risk behaviors of individuals before and after the intervention. Moreover, cost data are not typically collected alongside HIV prevention programs. An overview of published research on modeling cost-effectiveness of HIV prevention programs is provided, a list of the important outcomes is presented and methodological issues associated with modeling such programs are discussed in this review. PMID- 19807452 TI - Cost-effectiveness of left ventricular assist devices. AB - Implantable left ventricular assist devices are expensive both in terms of the actual device cost and postoperative care. These devices have been successfully used in numerous life saving indications but their cost and cost efficacy remains unclear. This report reviews these issues focusing on device, in-hospital, and posthospitalization costs. PMID- 19807453 TI - Cost-effectiveness of emergency contraceptive pills in the public sector in the USA. AB - Emergency contraception, which prevents pregnancy after unprotected sexual intercourse, has the potential to significantly reduce the incidence of unintended pregnancy and the consequent need for abortion. The cost-effectiveness of emergency contraceptive pills in the public sector in the USA is determined. The cost-savings when a single emergency contraceptive treatment is provided following unprotected intercourse are modeled along with when women are provided emergency contraceptive pills in advance. This resulted in a saving of USD 1.94 4.47 for each dollar spent on a single treatment of emergency contraceptive pills depending on the emergency contraceptive pill regimen and assumptions about savings from costs avoided by preventing mistimed births. The dedicated products Preven and Plan B are cost-effective even under the least favorable assumption that mistimed births, when prevented today, occur 2 years later. Each dollar spent on advance provision of Preven saves from USD 2.06 to 20.05, and each dollar spent on Plan B saves from USD 1.51 to 9.30, depending on the regular ongoing method of contraception, how consistently emergency contraceptive pills are used when needed, and whether mistimed births are averted forever or simply delayed. Emergency contraception is cost-effective whether provided when the emergency arises or in advance to be used as needed. More extensive use of emergency contraception could save considerable medical and social costs by reducing unintended pregnancies, which are expensive. PMID- 19807454 TI - Linking pharmacoeconomic analyses to results of systematic review and meta analysis. AB - Pharmacoeconomic analysis applies quantitative modeling to the assessment of the clinical and economic impact of new drugs. Users of pharmacoeconomic analysis include government agencies, government payers and policy makers, private payers (including managed care organizations) and pharmaceutical companies. Pharmacoeconomic analyses can aid policy decisions, provide support for better allocation of scarce resources and assist clinical decisions. Since pharmacoeconomic analyses can have a wide impact, it is important that they are based upon reliable data. Well-conducted systematic reviews and meta-analyses can provide high quality data to pharmacoeconomic analyses, with considerable synergy achieved by combining these two powerful methodologies. An overview of systematic review and meta-analysis are presented and some examples of their use in pharmacoeconomics are described. PMID- 19807455 TI - Compliance with inhalatory therapy: an increasingly recognized clinical outcome. AB - Compliance with medications is one of the major issues encountered in medical practice in the treatment of many chronic diseases, as it represents the interface between the effectiveness of the drug regimen and management of the disease. Noncompliance with inhalatory therapy could be the leading cause of poor disease control and increased morbidity in conditions, such as asthma or chronic obstructive pulmonary disease. The strongest predictors of compliance are represented by the marital status, age and education of the patient, however, more research is needed in this field. Psychological factors are also known to play a role, as does the medication regimen. Assessment methods most commonly used for inhalatory therapy are self-reporting and canister weight but electronic diaries and electronic monitoring devices are more promising and reliable. Educational interventions and disease self-management plans are known to be effective in improving compliance but more specific strategies must be established. Special attention is required in children and the elderly since the compliance-related issues in these populations are more complex. PMID- 19807456 TI - Tobacco control policies and reductions in smoking rates and smoking-related deaths. AB - Tobacco control policies are examined through a simulation model projecting smoking prevalence and associated premature mortality over a 40-year period beginning in the year 2000. The basic model and policy inputs are described. Results of each of six types of policies (tax, clean air, mass media, advertising, cessation and youth access) are examined independently and as a package. These results are presented in comparison with a business as usual scenario, which assumes no significant policy developments as of the year 2000. Results suggest that the largest potential gains come from implementing a comprehensive tobacco control policy package but that significant inroads to reducing smoking prevalence and premature mortality may be achieved through tax increases and clean air laws. PMID- 19807457 TI - Cost per quality-adjusted life year and disability-adjusted life years: the need for a new paradigm. AB - Two different paradigms have been proposed for setting priorities for access to healthcare: cost per quality-adjusted life year based on interventions, and disability-adjusted life years based on the burden of disease in a population. These formal paradigms make explicit the assumptions made implicitly every day in delivering and hence rationing access to healthcare. This paper outlines each paradigm's methodological problems and argues that each paradigm is incomplete in terms of providing the information necessary for making budgetary decisions on healthcare. It argues that a scientific revolution is required to create a new paradigm by combining the strengths of each. PMID- 19807458 TI - Quality of life assessment in cancer clinical research: current status and a look to the future. AB - Quality of life (QOL) is a term that has been used to denote outcomes as experienced by the patient. Although definitions of QOL vary, virtually all investigators agree that QOL in clinically-ill individuals encompasses multiple domains or areas of well-being (including, at a minimum, physical, psychological and social functioning, as well as symptoms) and that the perspective of the patient is critical in any measurement of QOL. While patient perspectives are important in any health condition, they become particularly so in diseases which are chronic or incurable and for which treatments have toxic or long-lasting consequences. Cancer fits these criteria. Cancer is the second leading cause of death in the USA, with one in every four deaths due to the disease. Approximately 38% of cancer patients die within 5 years of diagnosis. Cancer treatments are powerful and toxic. For example, surgery removes tumors and in many cases, adjacent body parts. Radiation therapy kills cancer cells by directing high energy x-rays, electron beams or radioactive isotopes at tumors. Chemotherapy causes the death of cancer cells through the administration of agents that disrupt a cancer cell's growth cycle. All of these treatments affect normal tissue, as well as cancer cells, and generally result in multiple side effects, such as loss of hair, nausea and frequently vomiting which are associated with chemotherapy. Treatment-related effects may be short-term and time-limited, chronic and persistent or late that only emerge after therapy has been completed, sometimes many years later. Cancer therapy involves continual efforts to minimize side effects while at the same increasing the chances of surviving the disease. As such, patient perspectives become critical in evaluating whether or not therapies and their toxicities are tolerable and acceptable. PMID- 19807459 TI - External beam radiotherapy in prostate cancer patients: short- and long-term health-related quality of life issues. AB - Health-related quality of life is an increasingly important issue in the treatment of prostate cancer. External beam radiotherapy is a frequently-used treatment option which can have short- and long-term side effects influencing the patients' quality of life. In this review the urinary, intestinal and sexual health-related quality of life issues related to treatment with external beam radiotherapy are addressed. Recent research reports allow us to provide an overview on both the short- and long-term of a patient's cancer experience. External beam radiotherapy has different effects on bowel, bladder and sexual function. This review suggests that external beam radiotherapy has a considerable short-term effect on bowel and sexual functioning, and that urinary function is rarely affected. In the long-term there is still a small, but non-negligible, effect on bowel functioning. Sexual function is greatly affected by external beam radiotherapy in the long-term. Recent radiotherapy treatment techniques, such as conformal external beam radiotherapy, tend to give better health-related quality of life outcomes. However, this needs confirmation by more studies in the short- and long-term including health-related quality of life measurements. PMID- 19807460 TI - Radiotherapy cost-calculation and its impact on capacity planning. AB - The rapid rise in health care expenses has resulted in an increased interest in the cost of treatments from a cost-effectiveness point of view for management purposes and in a reimbursement setting. The economics of radiotherapy within the global context of health care, and more specifically of cancer therapy, are discussed in this review. Furthermore, the calculation of radiotherapy costs from an institutional perspective using activity-based costing and on capacity planning in radiotherapy - at the departmental as well as at the national level - by integrating cost, epidemiological and scientifico-technological data are focused on. PMID- 19807461 TI - Meeting the US FDA's evidence standard with health-related quality of life claims. PMID- 19807463 TI - First international conference on economics and human biology. PMID- 19807464 TI - Fluticasone propionate in COPD airway inflammation: there is a partial effect. AB - Chronic obstructive pulmonary disease is a mainly tobacco-related disease currently representing the fourth highest cause of mortality worldwide. It is characterized by progressive chronic airway obstruction, the underlying pathogenic mechanism being inflammatory. Currently, apart from neutrophils and macrophages, there is an evolving role of T-lymphocytes, especially in smokers who develop chronic obstructive pulmonary disease. Oral steroids produce a clinical improvement in approximately 10% of chronic obstructive pulmonary disease patients. Inhaled steroids, such as fluticasone propionate, failed to stop lung function deterioration, but reduced the health status decline and number of exacerbations. Its effect on the inflammatory process was analysed in the study authored by Hattotuwa et al. The primary outcome was the effect of fluticasone propionate on the inflammatory cells present in the bronchial biopsies. Secondary outcomes were the symptoms, lung function decline and number of exacerbations. Fluticasone propionate had a partial effect on inflammation did not influence the progressive airway obstruction, partially improved the symptoms and reduced the number of exacerbations. PMID- 19807465 TI - Cost allocation in antihypertensive drug therapies. AB - The objective of this paper is to evaluate persistence with treatment and resources allocation in antihypertensive pharmacotherapy in a 'real world' population. An administrative database listing all purchased drugs was used to perform a longitudinal analysis. The study included all new users over 20 years of age receiving a first prescription for amlodipine, atenolol, fosinopril, indapamide, or losartan, in an enrolment period of 12 months. The follow-up period lasted 12 months. According to prescriptions dynamics, subjects were classified as same therapy, combination, switching, interruption and occasional utilization. The 34.9% study cohort, persisted with treatment (21.1% on same therapy, 4.7% on combination and 9.1% on switching), while 65.1% did not persist (10.9% on interruption and 54.2% on occasional use). The overall drug cost accounted for persistent (69.0%) and nonpersistent subjects (31.0%). The annual average cost ranged from euro32.80 for occasional users to euro274.69 for those in combination. In clinical practice, a high percentage of patients do not receive adequate antihypertensive therapy, since the 65.1% of subjects did not persist with treatment. This results in a level of pharmaceutical expenditure that cannot be considered appropriately allocated. PMID- 19807466 TI - Outcomes of dalteparin use following total knee arthroplasty: a retrospective analysis. AB - In this study, we assessed the clinical and economic consequences of using dalteparin as first-line prophylaxis for deep-vein thrombosis and pulmonary embolism in patients undergoing inpatient rehabilitation following total knee arthroplasty. The objective of the study was to demonstrate that dalteparin can be used effectively and safely in this indication. Using data abstracted from medical records of patients admitted to an Oklahoma rehabilitation center following total knee arthroplasty, we found the occurrence of thromboembolic and bleeding events in patients receiving dalteparin to be low. Results of this study also suggest that use of dalteparin in this indication could offer rehabilitation hospitals a valuable source of cost savings, without compromising patient care. PMID- 19807467 TI - Incidence versus prevalence modeling in pharmacoeconomics. AB - Increasing standardization in pharmacoeconomics calls for systematic use of epidemiological measures, such as incidence and prevalence estimates. The implications are relevant to predicting illness events and costs and to formulary decision-making. The choice of method is determined by the purpose of the study and the nature of the disease. Prevalence-based costs are assigned to the years in which they occur - results are relevant for budget impact, cost control and in chronic conditions. In contrast, the incidence-based approach assigns the stream of costs to the year it starts. It requires knowledge of the disease course, survival rates and the impact of the illness on lifetime earnings. It is relevant for making decisions among alternative treatments,short-term cost-effectiveness and acute diseases. PMID- 19807468 TI - The use of modeling in the economic evaluation of vaccines. AB - As a consequence of the increased role of pharmacoeconomics in policy-making, economic evaluations are performed at more and more early stages in the development of a therapeutic. This implies the development of models to assess the future impact of an intervention and to account for the level of uncertainty in the associated parameters. This also applies for economic evaluations of vaccines, where not only progression of disease and associated costs are important, but the transmission of the causing agent in the target population also has to be modelled. In this review, we provide an overview of the models that have been used in recent publications on the pharmacoeconomics of vaccines and go deeper into some of the methodological issues associated with the use of models in the economic evaluation of vaccines. PMID- 19807469 TI - Cost-effectiveness of cognitive-behavioral therapy for depression: current evidence and future research priorities. AB - Depression is a common psychiatric disorder. Although a variety of antidepressant medications have been shown to be effective, psychological therapies are highly valued by patients and some (such as cognitive-behavioral therapy) may be as clinically effective as antidepressants. Given similar clinical effectiveness, issues of differences in costs between cognitive-behavioral therapy, medication treatments and other approaches to mental healthcare are important. The current article discusses issues relating to the evaluation of the cost-effectiveness of cognitive-behavioral therapy for depression, reviews current empirical evidence concerning cost-effectiveness and highlights key issues for future research. PMID- 19807470 TI - Measuring the costs and benefits of pharmaceutical expenditures. AB - In response to concerns about the cost burden of medical care in general and rising pharmaceutical costs in particular, a number of recent studies have examined trends in pharmaceutical expenditures. These studies have reached apparently disparate conclusions about factors responsible for rising pharmaceutical costs and the role that price and volume increases have played. In this paper, we examine methodological approaches that have been used to describe and analyze trends in medical and pharmaceutical expenditures using recent studies as examples. We present in detail one methodology for decomposing the factors of pharmaceutical spending and describe how variation in method can affect study findings. Finally, we consider some research extensions that we feel could considerably improve our understanding of the costs and benefits associated with pharmaceutical expenditures. PMID- 19807471 TI - Pooling multisite administrative data for economic analysis. AB - Economic evaluations, such as cost-offset analyses, are receiving increased attention by US health insurers and payers. Administrative data collected by health maintenance organizations (HMOs) are considered an efficient source of utilization and cost measures for multisite economic analyses that increase external validity. However, pooling administrative data are problematic because HMO data sources reflect differences in systems of care, costing and coding. This paper describes issues arising from the pooling of HMO administrative cost data for use in multisite economic evaluations. We describe the attributes of administrative data that are relevant to costing and discuss their implications. We then briefly describe our experience with pooling these data, discuss lessons learned and offer suggestions for researchers working with such data. PMID- 19807472 TI - Publicly available US national surveys that capture lost productivity. AB - With the limited resources available, healthcare purchasers need to be able to determine the value of the pharmaceuticals or healthcare interventions that they purchase and their beneficiaries receive. From a purchaser's perspective, at least part of the benefit resulting from interventions may be the associated decreased lost productivity among their beneficiaries. However, the ability to value lost productivity at a population level is dependent on the availability of productivity data. This review provides a taxonomy of productivity measures within the currently available, national databases and surveys within the USA that capture lost productivity data. As the field of productivity measurement matures, with more precise data and methods to estimate patients' lost productivity in national survey data, healthcare employers and policy-makers will be able to make better-informed decisions concerning the medications that have an impact on the effects of diseases at a population level. PMID- 19807473 TI - Impact of psychological distress on outcomes in cancer patients. AB - Cancer is a major health problem and is the second leading cause of death in the USA. A diagnosis of cancer and cancer treatments are stressful events that can cause moderate-to-severe levels of emotional distress among some patients. This article provides an overview of the effects of psychological distress on various facets of patients' lives, including quality of life, health behaviors and biological health outcomes. Studies investigating the effectiveness of psychosocial interventions designed to reduce distress and improve psychological adjustment are also reviewed. We discuss methodological and conceptual weaknesses in study designs that may limit interpretation of study findings. Finally, we highlight key questions that remain to be addressed and propose several directions for future research efforts. PMID- 19807474 TI - Comparative policy analysis and health-system reform. PMID- 19807476 TI - Hospitalizations and mortality in the Lung Health Study. AB - The Lung Health Study previously reported that an aggressive smoking cessation intervention significantly reduced the decline of the forced expiratory volume in 1 sec in middle-aged patients without clinical symptoms, but with mild airway obstruction. This effect was the most evident during the first year after quitting smoking and moreover, persisted over the study period (5 years). In the current report, the impact of smoking cessation on mortality and morbidity was assessed in the same population. Being male, age, higher blood diastolic blood pressure were the main mortality risk-factors, whereas being female and alcohol consumption were risk-factors for subsequent respiratory disease. PMID- 19807477 TI - Pharmacoeconomics of antiviral therapies for Herpes zoster infections. AB - Herpes zoster is a common problem, particularly in the elderly, that can lead to postherpetic neuralgia - a significant source of suffering and diminished quality of life. Antiviral medications diminish acute and chronic symptoms of zoster, but whether the relatively high cost of these medications is worth their beneficial effects is controversial. This review updates our prior analysis, synthesizing evidence on the clinical features and costs of zoster and its sequellae, as well as the effects of antiviral therapy, using decision analysis techniques. We find antiviral therapy economically reasonable in immunocompetent adults for whom treatment is clinically recommended: severely symptomatic acute zoster in any adult and milder zoster in adults of 50-60 years or older. PMID- 19807478 TI - Caring for relatives with dementia: willingness-to-pay for a reduction in caregiver's burden. AB - Informal care constitutes a large percentage of total care for patients with dementia. Providing care for cognitively impaired individuals is stressful and demanding. This study applies the contingent valuation method to estimate the willingness-to-pay of informal caregivers for a reduction of their burden. In 109 face-to-face interviews, caregivers with demented relatives stated their willingness-to-pay for three hypothetical treatments of the patient. Results suggest that caregiver's disutility from burden is considerable. On average, caregivers are willing to pay 2200 Swiss Francs (US$1500) per year for a reduction of their burden level from moderate to low. Therefore, interventions which combat the negative consequences of caregiving have great potential to yield a social net benefit. PMID- 19807479 TI - Cost-effectiveness of olopatadine in seasonal allergic conjunctivitis treatment. AB - Cost-effectiveness of olopatadine was assessed through a literature search and available information. Seasonal allergic conjunctivitis is a nonlife threatening disease associated with increased health expenditures, decrease in quality of life and a large number of days off work and school. Few studies have been performed on the cost-effectiveness of compounds aimed at treating seasonal allergic conjunctivitis. Olopatadine is a new, dual-action compound indicated in the treatment of seasonal allergic conjunctivitis which has been found in clinical trials to be more potent and efficacious than currently available treatments. The rare cost-effectiveness studies performed on olopatadine in seasonal allergic conjunctivitis showed that given its higher efficacy, it could save around euro10 of relapse direct costs in a range of European settings. Further research is required in order to estimate precisely the cost effectiveness of this compound, particularly including indirect costs and in other countries. PMID- 19807480 TI - Adenosine and dipyridamole in nuclear imaging: pharmacoeconomic considerations. AB - Adenosine and dipyridamole are coronary vasodilators used in nuclear imaging. The acquisition cost of adenosine is four to ten times higher than dipyridamole. Data suggest comparable sensitivity, although specificity may be lower with dipyridamole. While side effect profiles are similar, fewer patients who receive adenosine require medical intervention to manage their side effects. Only three pharmacoeconomic evaluations comparing adenosine and dipyridamole are available - the data are limited by the methodology and study design. Two studies found that total costs with adenosine were lower compared with dipyridamole. In contrast, the interim results of an evaluation conducted with generic dipyridamole found that total costs were higher with adenosine than with dipyridamole - the final results are awaited. In addition, evaluations of downstream costs are needed to clarify the economic implications of a lower specificity with dipyridamole. PMID- 19807481 TI - Quality assessment of economic analyses of pharmacological and nutritional therapy for hyperlipidemia. AB - Our objective was to evaluate the quality of published pharmacoeconomic studies of lipid-lowering therapies. A 13-item evaluation checklist was used to assess the quality of articles. Two reviewers assessed each article. Discrepancies in ratings were resolved using a third reviewer. Potential scores on each item ranged from 0 to 4 and 50 articles met our inclusion criteria. Mean quality scores ranged from 1.73 to 3.89. Treatment and policy implications are: secondary prevention is more cost-effective than primary prevention, the cost-effectiveness of lipid-lowering treatments correlates with risk-factors and statin drugs are more cost-effective than cholesterol-sequestering agents. This review provides a summarization of the literature regarding the cost-effectiveness of lipid lowering therapies. Results from lower-quality studies may be less accurate because the most common criteria assessed as questionable or incorrect were perspective, measurement and analysis. PMID- 19807482 TI - Strategies for assessing health economic and quality of life outcomes in multiple sclerosis. AB - Multiple sclerosis is a disabling disease that, until recently, had few effective treatment options. With the advent of new medical technologies to treat multiple sclerosis has come the need to assess the long-term clinical, economic and quality of life trade-offs associated with these treatments to inform medical decision-making, as well as equitable resource allocation. Assessing these trade offs in multiple sclerosis is particularly challenging. Therefore, considering an appropriate evaluation strategy prior to initiating a health outcomes study in multiple sclerosis is essential. This review presents an overview of multiple sclerosis, the challenges facing researchers and offers strategies and a framework for assessing economic and quality of life outcomes in patients with multiple sclerosis. PMID- 19807483 TI - Infant, child and adolescent quality of life: surveys performed in a European context. AB - The authors have validated three quality of life questionnaires designed to assess mostly subjective aspects of quality of life, in infants (QUALIN questionnaire, a parents' report), children (AUQUEI questionnaire, a pictured self-report for children aged 3-11 years old) and in adolescents (OK.ado questionnaire). Psychometric properties being satisfactory, these questionnaires were completed by different samples, in different health states. We were thus able to detect the influence of several factors on children or adolescents' quality of life: illness tends to alter quality of life, but this is also the case for several psychosociological conditions, especially when combined with the child's age. In adolescents, one has to be careful in interpretation of results, since the healthy adolescent tends to complain to adults. PMID- 19807484 TI - Treatment satisfaction as an outcome measure in cancer clinical treatment trials. AB - For many years, medical research has focused on 'hard' end-points, such as survival or time to progression, when evaluating research in cancer clinical trials. However, over recent years more recognition has been given to 'softer' end-points, such as quality of life and treatment satisfaction. In many respects, research into treatment satisfaction is in its early stages of development, but we are already beginning to understand that patients' treatment satisfaction can provide great insights into the management of cancer patients. In our review, we discuss the specific conceptual and methodological issues we face when assessing treatment satisfaction in cancer clinical trials and critically review earlier studies of cancer treatment satisfaction. PMID- 19807485 TI - Preference assessment in patients with asthma. AB - Preference measurement is an important concept in health economics. The theory provides a common outcome measure that can be compared across diseases. Instruments used to measure preference, such as the standard gamble, time trade off, visual analog scale and multiattribute health state classification systems, have been used in asthma studies. However, preference measurement has not been routine in asthma studies, partially because of concerns about the sensitivity of the instruments in detecting important changes in asthma. The development of disease-specific, preference-based instruments may change that trend. However, such instruments move away from the goal of providing a single summary measure comparable across diseases. The instruments used to measure preference and preference results in asthma studies are reviewed in this paper. PMID- 19807486 TI - Therapeutic drug monitoring: is it cost-effective? AB - During the last 30 years, therapeutic drug monitoring has evolved from an abstract consideration to a routine intervention. In this increasingly cost conscious environment, questions are raised about the cost-effectiveness of therapeutic drug monitoring. This review presents the state of therapeutic drug monitoring today, predictions for its near future and the status of economic analyses of the intervention. PMID- 19807488 TI - Tailor-made or off-the-rack? The problem of transferability of health economic data. PMID- 19807487 TI - Update on effective interventions to increase tobacco cessation. AB - Tobacco use is the leading preventable cause of death in the USA and will soon be the leading cause of preventable death worldwide. The only way to decrease tobacco-related morbidity and mortality in the short term is to help current smokers quit. Fortunately, effective clinical- and population-based interventions that increase tobacco cessation exist. However, these interventions are not being implemented, leaving most smokers to use the least effective approach to cessation (an unassisted quit attempt). This review summarizes the evidence for the effectiveness of clinical- and population-based interventions and recommendations from various organizations regarding tobacco use treatment. It also outlines proposed strategies for improving clinical and public health practice to increase tobacco cessation. PMID- 19807490 TI - Methods and models in health-related quality of life (HRQOL) research: a state-of the-art review. PMID- 19807491 TI - Cost of diabetes-related complications: current issues, future directions. AB - The costs of diabetes are significant, approaching US$100 billion. Medical costs for patients with diabetes are between two- and four-times higher per patient per year when compared to persons without diabetes. Long-term complications of diabetes are likely to account for much of this excess. However, the total economic burden related to diabetes complications is not fully understood due to several factors. These include diagnostic, comorbidity and coding issues, as well as limitations in longitudinal data and the lack of information on the impact to work productivity. Future diabetes care and cost estimates will benefit from the use of electronic medical records. PMID- 19807492 TI - Managing diabetic foot ulcers: economic consequences in the USA. AB - In the USA, 11 million people are currently diagnosed with diabetes - poor glycemic control increases their risk of developing diabetes-related complications. Foot ulcers are one of the more common complications and patients developing foot ulcers are at a higher risk of having a lower extremity amputation. The majority of foot ulcers are managed in an outpatient setting, but hospitalization accounts for approximately three quarters of the total expenditures on medical care related to diabetic foot ulcers. A series of initiatives from the US government and healthcare providers aim to improve the management of diabetes, but more research is needed to establish which reduce the prevalence of foot ulcers and to establish the most cost-effective approach. PMID- 19807493 TI - Long-term social costs of hypertension. AB - According to published studies, much of the cost of hypertension is due to antihypertensive drug treatment. However, the cost of hypertension also includes the cost of an increased frequency of cardiovascular events when hypertension is not controlled. Although conceptually accepted by the scientific community, the achievement of appropriate blood pressure levels is less feasible than expected and studies demonstrate that only 13-27% of hypertensive patients are adequately informed, treated and controlled for their hypertension. This puts a tremendous burden on the healthcare system and society, since uncontrolled hypertension leads to higher rates of cardiovascular events and ultimately death. This paper demonstrates the paucity of reliable cost-of-illness estimates for the long-term consequences of uncontrolled hypertension and suggests that it is understandable that public and private payers focus on the immediate short-term costs of treating hypertension, paying less attention to potential cost savings of fewer cardiovascular events, as these costs are far less well defined. This paper also suggests that hypertension as a disease is an ideal candidate for disease management strategies and programs, as prevention of its long-term consequences should be the focus of medical treatment and could be better achieved through an integrated multispecialist and multisetting approach. PMID- 19807494 TI - Cost-effectiveness of nonemergency use of phenytoin. AB - Parenteral phenytoin is an effective agent used to manage seizures, but it is associated with adverse effects and must be given intravenously. Fosphenytoin is higher in drug cost, but is more soluble, better tolerated and can be infused at rates three times that of phenytoin. When infusion rate is not an issue the adverse effect risk becomes a focus of concern, as well as cost. The determination of the point at which the treatment costs of infusion-related adverse effects of phenytoin outweigh the drug costs of fosphenytoin has been attempted through various clinical economic analyses. Strategies developed to promote the safe use of intravenous phenytoin are based on patient selection, recommended administration methods and patient monitoring. When rapid attainment of serum phenytoin levels is required or in patients at high risk of adverse effects secondary to parenteral phenytoin, fosphenytoin is clearly preferred. This review will focus on the implications of the use of parenteral phenytoin products in the nonemergent setting where potential adverse effects of parenteral phenytoin may be avoided with use of established criteria for patient selection and administration. PMID- 19807495 TI - Cost-effectiveness of hepatitis C screening of blood donations. AB - Some screening programs have very high costs per life-year saved and require large financial support. Public policies decision-makers need criteria to appreciate the costs and benefices of such programs. The safety of blood transfusions is a specific example of such a situation, in which the decision to reduce residual risks could be a highly debatable use of resources when the additional effectiveness is small. This article analyzes various economic evaluations of hepatitis C virus screening strategies and discusses the results. The screening of blood donations for hepatitis C virus antibodies is cost effective and enzyme-linked immunoblot assay testing appears to be the most efficient and reliable test for daily processing of a large number of samples in a large blood bank. In contrast, new tests proposed to increase safety in blood donations, such as the adjunction of polymerase chain reaction testing, do not compare favorably with socially-acceptable threshold in health economics. PMID- 19807496 TI - Health status in COPD: current data and future trends. AB - In chronic obstructive pulmonary disease, health status is an important outcome for clinicians, the pharmaceutical industry and policy-makers. It has been assessed using both generic and disease-specific questionnaires. The generic questionnaire most frequently used in the context of chronic obstructive pulmonary disease is the Medical Outcome Study Questionnaire Short Form-36, whereas as the disease-specific Saint George's Respiratory Questionnaire has been used extensively to assess the effectiveness of pharmacological interventions, such as b2-agonists, theophylline, inhaled steroids and anticholinergics and the Chronic Respiratory Questionnaire has been proved to be a reliable instrument for the assessment of pulmonary rehabilitation programs. An interesting upcoming research direction would be the further involvement of health status in clinical decision-making processes. PMID- 19807497 TI - Measuring quality of life in rectal cancer patients. AB - Quality of life assessment in rectal cancer patients is necessary to provide a more comprehensive understanding of the outcome of surgery and other forms of treatment. Quality of life aspects must be placed in broader concept and should provide complementary information to healthcare professionals, together with classical outcomes, such as mortality, morbidity and long-term survival. There is a general lack of well-designed quality of life trials in rectal cancer patients. Ideally, individual patients could benefit from the data analysis, insofar as deficits should be disclosed and appropriately addressed. Finally, quality of life studies may also be used to monitor and evaluate the quality of care which patient receive, potentially leading to improved management. PMID- 19807498 TI - Quality of life in irritable bowel syndrome: measurement techniques and relevance of current knowledge. AB - Irritable bowel syndrome is characterized by abdominal pain and/or discomfort related to an abnormal bowel habit. Irritable bowel syndrome is very common in society and treatment alternatives are scarce. This disorder may be viewed as a benign disorder from a strictly medical point of view, but in many patients there is a chronic, relapsing course, which severely affects the activities of daily living. In recent years, interest in quality of life in irritable bowel syndrome has increased dramatically, both regarding comparisons with the normal population and other disorders and detecting changes due to different treatment regimes. This review presents the concept of quality of life and instruments used to assess quality of life in irritable bowel syndrome. It also reviews the current knowledge of quality of life in irritable bowel syndrome, based on existing studies. PMID- 19807499 TI - Health-related quality of life assessment in chronic kidney disease. AB - Chronic kidney disease affects over ten million Americans. Most severely affected patients undergo dialysis or transplantation at high economic and human cost. Each year, 70,000 Americans die from causes related to kidney failure. Health related quality of life has been an important area of investigation. Recent advances include the development of new instruments that better distinguish among treatment modalities and differing patient populations. This article provides a brief state-of-the-art review of health-related quality of life instruments in chronic kidney disease, describes issues affecting patients, factors associated with decreased health-related quality of life and summarizes studies of health related quality of life in this population. It concludes with our recommendations for further work and a look at what the future might hold. PMID- 19807500 TI - Why study pharmacoeconomics? PMID- 19807502 TI - Using economic evidence in reimbursement decisions for health technologies: experience of 4 countries. PMID- 19807503 TI - Secondary prevention fallacy: pitfalls in comparing with primary. AB - A basic tenet of our culture is the idea that preventing ills is better than curing them. This principle is entrenched in many proverbs and popular admonitions: 'A stitch in time saves nine', 'An ounce of prevention is worth a pound of cure'. These well-worn sayings crisply convey our sense that it is preferable to intervene before a bad event occurs than to wait and try to mitigate it afterwards. Yet, economic analyses have demonstrated that in healthcare this does not hold: 'primary prevention' does not provide as good value-for-money as 'secondary prevention'. Is this reversal of folk wisdom correct? Or, instead, is it a result of faults in the application of the relatively new methodology of cost-effectiveness analyses? PMID- 19807504 TI - Pharmacoeconomics of Helicobacter pylori: eradication versus maintenance therapy in controlling peptic ulcer disease. AB - We performed an electronic search in Medline and EMBASE for papers comparing Helicobacter pylori eradication and H(2)RA-maintenance therapy in peptic ulcer. Treatment to eradicate H. pylori in patients with a proven ulcer has a favorable cost-effectiveness compared with maintenance therapy, with benefits for the patient and society. In most studies, it was assumed that no more gastrointestinal drugs were needed after eradication. Additional research is needed to provide empirical evidence on gastrointestinal drug utilization after eradication of H. pylori. The development of a vaccine against H. pylori remains however the ultimate goal in the fight against peptic ulcer disease. PMID- 19807505 TI - Quantifying stochastic uncertainty and presenting results of cost-effectiveness analyses. AB - In the last decade, major advances have been made in the statistical methods for quantifying uncertainty in stochastic cost-effectiveness studies. In this paper, we provide a guide to the literature in which we highlight the preferred methods for confidence interval estimation, new developments in the formulation of the cost-effectiveness problem, suggested ways for presenting results and the areas in which future research may develop. The overall approach taken is nontechnical, with an emphasis on graphical rather than algebraic presentation of methods. PMID- 19807506 TI - Incremental cost-effectiveness ratio and incremental net-health benefit: two sides of the same coin. AB - In recent years, an alternative framework for cost-effectiveness analyses has been growing in popularity. Instead of the incremental cost-effectiveness ratio for which statistical inference is often difficult, the incremental net-health benefit (INHB), a linear transformation of incremental costs and effectiveness, has been utilized. The linear structure of this statistic allows easy computation and interpretation of confidence intervals, hypothesis tests and acceptability curves. It is often difficult, however, to switch decision-making procedures without first verifying the appropriateness of the new methods. In this paper, we demonstrate the decision-making similarities between the INHB and the incremental cost-effectiveness ratio and describe how the INHB can be used to clarify inference of the incremental cost-effectiveness ratio. We also describe the two statistics in terms of the DeltaE-DeltaC plane, thus allowing both a mathematical and graphical comparison of these similarities. We conclude with a general discussion of cost-effectiveness analyses and advocate Bayesian, rather than frequentist inference as the more intuitive and powerful decision-making framework. PMID- 19807507 TI - Economic evaluations of adult pneumococcal vaccination strategies. AB - We performed an extensive search for full economic evaluations on pneumococcal vaccination in adults. We included 11 North American and 9 European studies for review. There was a clear preference for cost-effectiveness and cost-utility versus cost-benefit analysis. In general, pneumococcal vaccination of the elderly <75 years and of particular risk groups is found to be relatively cost-effective and potentially cost-saving to the health care sector and to society. However, there is substantial uncertainty about the effectiveness of the intervention in these target groups. From 1997 on most analysts have taken this partly into account by distinguishing between invasive and non-invasive pneumococcal disease. In order to be relevant, future analyses should use quality adjusted life-years, adapt life-expectancy for people at risk of infection, narrow the targeted age groups, include realistic programme costs and discuss the potential influence of unrelated future costs. Finally, estimating the impact of antimicrobial resistance (not included in any of the analyses) seems particularly challenging for the future. PMID- 19807508 TI - Economics of fall prevention programs: evidence and research priorities. AB - Injurious falls, particularly among the elderly, are a leading cause of morbidity, mortality and health and social care expenditure. Non injurious falls may result in significant quality of life losses through generating a fear of subsequent falls. Falls may be due to intrinsic (age-related physiological changes, diseases or polypharmacy) or extrinsic (environmental hazards and behavior) factors. Although interventions target both of these, much of the focus is upon extrinsic factors, through changes to behavior and the environment. This paper reviews the cost-effectiveness of such interventions in reducing the risk of new falls, or modifying the harm caused in the event of a fall, for the general unselected population of the elderly living independently in the community. PMID- 19807509 TI - Cost of depression: current assessment and future directions. AB - Depression is a common psychiatric disorder affecting approximately 17 million Americans each year and resulting in a significant economic burden, estimated at $43-$53 billion in 1990. The cost burden of depression extends beyond the direct cost of treatment, to include the costs of lost productivity, both while at work and days absent from work, as well as lost earnings due to increased mortality and the impacts experienced by a patient's caregivers. This range of costs is discussed, in the context of the prevalence and impacts of depression and detailing the cost components of depression. We highlight that existing estimates of the cost of depression are underestimates and we conclude with areas for future research. PMID- 19807510 TI - Pharmacoeconomic evaluation of intensified antiretroviral treatment strategies in HIV/AIDS. AB - There have been great technological advances in the use of antiretroviral therapies to slow down disease progression in HIV/AIDS. Combinations of therapeutics and the use of several diagnostic methods have resulted in both declines in mortality and the occurrence of opportunistic infections. The higher costs of these therapeutics have prompted questions about the economic aspects of treatment with antiretrovirals. In this review, we provide an overview of the research that has been published on this topic and list the important outcomes and methodological issues associated with the different therapies. PMID- 19807511 TI - Severity of illness versus expected benefit in societal evaluation of healthcare interventions. AB - Society's valuation of a healthcare outcome depends not only on the size of the gain in well-being (utility), but also on the severity of the initial condition. This seems to be a major problem with the conventional utility-based QALY approach to outcome evaluation. In particular, QALY calculations based on utilities from multiattribute utility instruments assign too high value to interventions for people with mild and moderate health problems compared with interventions for people with severe and life threatening diseases. Analysts should somehow make corrections for this bias in economic evaluations of healthcare programs. PMID- 19807512 TI - Quality of life issues in gynecological cancer. AB - Understanding quality of life has become an increasingly important issue in the treatment and clinical care of patients with gynecological cancer. The short- and long-term side-effects of treatment may also impact on a woman's self worth and sexuality. In our review we address the key issues related to the treatment and management of patients with gynecological cancer. We focus on the methodology issues and limitations in the design of studies and propose our view of how to approach and enhance better quality trials in the future. PMID- 19807513 TI - Health-related quality of life in Parkinson's disease: outcomes of the therapeutic interventions. AB - Main reasons for measuring health-related quality of life (QoL) are: 1) the unique information supplied by QoL measures, 2) the relationship between perceived health and use of resources and 3) the usefulness for evaluation of outcome and decision making. As Parkinson's disease impacts strongly on the life of patients, it is important to assess the effect of the interventions not only on clinical aspects, but also from the point of view of the perceived health state. The QoL conceptual framework, the type of tools for measuring QoL and the trials that applied QoL measures as outcome variables in PD are reviewed in this article. PMID- 19807514 TI - Medical reimbursement for diagnostic tests. PMID- 19807516 TI - Economic benefits of pioglitazone for treating patients with Type 2 diabetes. AB - Diabetes remains a significant economic burden to national healthcare systems. The traditional oral agents used to treat Type 2 diabetes do not address the underlying insulin resistance responsible for the development of diabetes. Newer medications, such as the thiazolidinediones, have been shown to reverse some of the metabolic processes believed to be responsible for the development of insulin resistance and ultimately, Type 2 diabetes. A comprehensive economic evaluation of pioglitazone using a modelling approach indicates that pioglitazone is a cost effective therapy for patients with Type 2 diabetes when used in combination with either a sulfonylurea or metformin. This drug profile analyzes the clinical data on the use of piogltiazone for the treatment of Type 2 diabetes and the various economic evaluations of pioglitazone in the literature. PMID- 19807517 TI - Levetiracetam: an innovative and cost-effective add-on drug for refractory partial epilepsy. AB - Poorly-controlled epilepsy can have a significant negative impact on quality of life, clinically important changes are seen in patients emotional well-being, cognitive functioning, social functioning and energy/fatigue levels. Poorly controlled epilepsy places an undue economic burden on the patient and community. Increased costs are seen in both direct and indirect healthcare costs (e.g., inpatient care and loss of earnings associated with time lost from work). Therefore, long-term efficacy and tolerability are key considerations when designing the patient's treatment regimen. Therapy can be individualized using both classical drugs and newer antiepileptics such as levetiracetam (Keppra, UCB Pharma Inc.), which is currently recommended as add-on therapy for partial-onset seizures. Studies have revealed characteristics that suggest levetiracetam is the first of a new class of antiepileptic drugs, differentiated by its innovative mechanism of action. Its efficacy and tolerability have enabled many patients who were refractory to treatment with other antiepileptic drugs to achieve long-term seizure freedom. Levetiracetam has a high long-term retention rate, a powerful measure of adverse events and efficacy over time. Another equally important benefit is ease of use, levetiracetam is administered twice-daily and has a simple titration regimen. Pharmacoeconomic data show that the incremental cost of treating patients with levetiracetam is low when compared with the benefits of seizure freedom. Ongoing studies suggest that this antiepileptic drug has potential as first-line treatment for many types of epilepsy and in many different patient populations. PMID- 19807518 TI - Methylnaltrexone and alvimopan: economic management of opioid-induced bowel dysfunction. AB - The occurrence of opioid-induced bowel dysfunction is common in the postoperative period and in chronic pain conditions. This debilitating adverse effect involves the activation of peripheral opioid receptors causing delayed recovery of normal colonic motility, prolonged postoperative ileus and increased morbidity. This translates into increased lengths of hospitalization and significant added attendant healthcare costs. Methylnaltrexone and alvimopan (Entereg, Eli Lily & Co.) are the first selective antagonists of gastrointestinal opioid receptors. Recent studies have shown their ability to reverse opioid-induced bowel dysfunction without reversing analgesia or precipitating CNS withdrawal signs in nonsurgical chronic pain patients receiving opioids. In addition, clinical studies with alvimopan suggest that it may normalize bowel function without blocking opioid analgesia in abdominal laparotomy patients with opioid-related postoperative ileus. These drugs may have clinical utility with an economic impact in managing opioid-induced bowel dysfunction. PMID- 19807519 TI - Patient recall and recall bias of health state and health status. AB - The reliability of recall patient reported outcomes, such as health-related quality of life, satisfaction and symptoms, varies substantially. The objectives of this special report are to identify key issues to consider in study design and provide suggestions for minimizing bias in studies including patient reported outcomes. A MEDLINE search identified several areas in which patient recall is subject to bias. Concordance between patient recall and baseline assessments (e.g., prior to an event or medical intervention) for these patient reported outcomes varies depending on the event being recalled, time since the event, and patient clinical and demographic characteristics. Symptom recall tends to be better than recall of health-related quality of life or pain intensity. Specific questionnaire techniques may help minimize the impact of recall bias. Further research is required to determine what factors or patient characteristics predict improved recall and what techniques minimize recall bias. PMID- 19807520 TI - Threshold denial rates in prior authorization prescription programs. AB - In this special report, the economics of prior authorization is examined with a focus on the break-even rate that requests are denied. Using a simple theoretical model, comparative static results are derived and used to consider the cost effectiveness implications of future changes in prior authorization policies. PMID- 19807521 TI - Memorial symptom assessment scale. AB - Patients with advanced illnesses often have multiple symptoms. As interest in palliative care and interventions for symptom control increase, the ability to assess multiple symptoms has become more important. A number of instruments have been developed to meet this need in cancer patients. This article reviews the development and applications of a multidimensional instrument, the Memorial Symptom Assessment Scale. The Memorial Symptom Assessment Scale has 32 symptoms and three dimensions of frequency, severity, and distress. Shorter versions - The Memorial Symptom Assessment Scale Short Form (32 symptoms with one dimension) and the Condensed Memorial Symptom Assessment Scale (14 symptoms with one dimension), and a version for children aged 7-12 years, have also been developed. A distinctive feature is the summary subscales for physical distress, psychological distress, and The Global Distress Index. The Memorial Symptom Assessment Scale has proven useful in description of symptom epidemiology, the role of symptoms in pain, fatigue, and spirituality; as a predictor of survival, and in proxy assessments of pain. The Memorial Symptom Assessment Scale has been used in studies of cancer and AIDS patients, and patients with advanced medical illnesses. Possible future roles of instruments such as the Memorial Symptom Assessment Scale include use in clinical trials, for pharmacoeconomic analyses, definition of symptom clusters and symptom burden, the development of symptom outcome measures, symptom monitoring, and improving care for patients. Continued research is needed for the versions of the Memorial Symptom Assessment Scale and other symptom instruments in different populations and applications. PMID- 19807522 TI - Evaluating the true cost of hypertension management: evidence from actual practice. AB - Hypertension is a prevalent and costly disease and drug cost is the most significant driver of total expense. Therefore, medications such as diuretics, with lower acquisition costs, and equal or better trial-based efficacy versus other classes of antihypertensives, are currently recommended as first-line therapy. However, observational data from actual practice suggest that antihypertensive drug acquisition costs alone are a poor predictor of total treatment cost. This review explores other important determinants of cost which must be considered, such as therapeutic turbulence and persistence on therapy, which cannot be measured with validity within a clinical trial environment. Actual practice data reveal that greater turbulence and poorer persistence is associated with older agents such as diuretics, versus newer, more tolerable medications. On the basis of observational evidence, the gap in total treatment cost associated with older versus newer antihypertensives is significantly less than that which is commonly reported and used as an argument for first-line treatment with diuretics. Continued constructive debate over the implications of observational data for the selection of a first-line antihypertensive therapy is warranted. PMID- 19807523 TI - Principles of good modeling practice in healthcare cost-effectiveness studies. AB - Decision analytic models are increasingly being used to present information on the costs and effects of both new and existing healthcare technologies. However, despite this increase, the literature on what constitutes 'quality' or 'good practice' in modeling is sparse, confusing and often conflicting. As a result there is a need to summarize these quality assurance and good practice principles into a framework that is useful for modelers and users of models alike. This review has attempted to summarize these principles into five broad categories that will assist in assessing whether a model should be considered 'SAVED' (has structural integrity, uses appropriate input data and calculation methods, validates the model output, has extensive use and reporting of sensitivity analysis, and if there is detailed and unbiased reporting and interpretation of study findings). These principles span every aspect of the cost-effectiveness analysis from model conception, development and calculation, to presentation and interpretation of the results. Modelers are strongly encouraged to actively consider these principles throughout the entire process of model development, analysis and write up. Users of modeling studies should be familiar with these principles in order to correctly appraise studies for their applicability, validity and interpretability. PMID- 19807524 TI - Socioeconomic and health benefits of lactoceuticals. AB - The rapid increase of healthcare and medication costs is a growing problem for industrialized countries. Several factors contribute to this problem and solutions are imperative to help the situation. In this review, the concept of lactoceuticals is introduced and why this class of product could have an impact in reducing healthcare and medication costs is explained. The term 'lactoceuticals' is defined and the key players already involved in this field are presented. This review will also summarize and list lactoceuticals that have reached a certain market maturity, and innovative lactoceuticals that will soon appear on the market following the correct clinical investigations and regulatory homologations. PMID- 19807525 TI - Health-related quality of life in prenatal diagnosis. AB - The objectives of this review include the conceptualization of the health-related quality of life effects of prenatal diagnosis and a brief summary of evidence on the short- and long-term effects of prenatal diagnosis on the health-related quality of life effects associated with chorionic villi sampling and genetic amniocentesis and the identification of important unresolved issues. Although this is not a systematic review, it is an update of published research on the utility approach to assessing the health-related quality of life in prenatal diagnosis. It is based on a search of publications by investigators known to be active in the area and a hand search of selected specialized journals. Important health states associated with prenatal diagnosis include both process (undergoing testing) and outcome. Empirical studies providing preference scores for health states associated with prenatal diagnosis highlight the importance of long-term outcomes relative to process. On average with respect to process, chorionic villi sampling is less burdensome than genetic amniocentesis. On average with respect to infrequent but potentially important outcomes as health states associated with diagnostic inaccuracy, genetic amniocentesis is less burdensome than chorionic villi sampling. Almost all of the existing evidence on the health-related quality of life effects of prenatal diagnosis reports on the experience of the women undergoing prenatal diagnosis. The preferences of partners have not been assessed. Furthermore, few studies have investigated subsequent reproductive behavior. Finally, there is considerable scope for the use of preference elicitation techniques in helping couples to decide on whether or not to undergo prenatal diagnosis and if they do, help them to choose the modality that best suites their preferences. PMID- 19807526 TI - Quality of life issues in non-small cell lung cancer. AB - For most patients with non-small cell lung cancer, maintaining or improving the quality of their remaining life may be as (or more) important than a small extension of survival. Thus, the assessment of quality of life should be a key component of treatment trials to inform future clinical decision-making. However, only a small percentage of randomized trials in non-small cell lung cancer have included quality of life and many of these may be criticized for poor design, inadequate sample size and/or complex analyses. A radical rethink over the way in which quality of life is included in trials is required. Much larger sample sizes, predefined quality of life hypotheses, innovative methods to improve compliance rates and standard methods of analysis are required. Until trialists begin to take note of these messages, information on quality of life from trials will continue to be largely distrusted and disregarded by doctors and patients. PMID- 19807527 TI - Use of the Greek McGill Pain Questionnaire in cancer patients. AB - Cancer directly causes pain and raises a significant clinical problem. Pain is the most dominant cause of suffering in cancer patients. The accurate assessment of pain is a prerequisite for its effective relief, and therefore, a universal methodology for the assessment of the pain status of a patient is required. There are many different methods and various instruments for the measurement of pain. The McGill Pain Questionnaire is a multidimensional instrument for the assessment of pain intensity and the multiple dimensions of the pain experience. Several translations and version of the McGill Pain Questionnaire exist. The Greek version was developed in Athens and has been tested for its use in cancer patients receiving palliative care. The Greek McGill Pain Questionnaire validation study provided preliminary evidence of the validity and reliability of the questionnaire. Future studies are needed to confirm the validity of the instrument in order to render it fully utilizable for the assessment of pain caused by the diverse conditions experienced by Greek patients. PMID- 19807528 TI - Radiotherapeutic treatment of Peyronie s disease. AB - Peyronie's disease is a benign condition, affecting middle-aged men of between 40 and 60 years of age. The symptoms are penile pain in the early inflammatory phase, curvature during erection and erectile dysfunction. The origin of Peyronie's disease is not fully understood. A trauma of the penis with subsequent deposition of fibrin and formation of a fibrotic plaque in the tunica albuginea is the most likely pathophysiological mechanism. Empirical treatments have been attempted with inconclusive results since most of the studies were noncontrolled. In this review, the most important data on the epidemiology, etiology, diagnostic approaches and treatment modalities of Peyronie's disease are addressed. Low-dose (12-15 Gy) radiotherapy has been reported as a successful treatment modality to reduce penile pain and curvature in the early stages of the disease, in noncontrolled studies. Currently, no side effects have been reported. This review critically addresses Peyronie's disease, its treatments, corresponding outcomes and future treatment possibilities. The problems of obtaining good-quality outcomes data for radiotherapeutic approaches are highlighted. PMID- 19807529 TI - Cost studies in clinical trials. PMID- 19807530 TI - Formulary decision methodology in the context of health system costs and insurance trends. PMID- 19807531 TI - Four quadrants of the cost-effectiveness plane: some considerations on the south west quadrant. PMID- 19807533 TI - 5th European conference on health economics. PMID- 19807534 TI - 10th Annual Australian Health Outcomes Conference 2004: 10 years on and going strong. PMID- 19807535 TI - Cost effectiveness of leflunomide in the treatment of rheumatoid arthritis in Japan. AB - Macro-level economic evaluations of drugs are expected to provide useful information for making the decision of whether to approve a newly developed drug as a first-line treatment. Leflunomide has been hoped to be an alternative to existing disease-modifying antirheumatic drugs, but has also raised questions about its cost effectiveness due to the high drug cost and high percentage of withdrawals due to adverse events. A method of cost-effective analysis using disability-adjusted life years, which makes it possible to assess the cost effectiveness of a drug at the macro level, is proposed. Using the proposed method, the cost effectiveness of leflunomide in the treatment of rheumatoid arthritis in Japan is determined. PMID- 19807536 TI - Cost effectiveness of methylphenidate versus AMP/DEX mixed salts for the first line treatment of ADHD. AB - Although it is widely accepted that stimulants are used as the treatment of choice in attention deficit hyperactivity disorder, many decision makers do not have appropriate information regarding an optimal first-line agent in treating this patient population. The cost effectiveness in choosing methylphenidate (Ritalin) or amphetamine/ dextroamphetamine mixed salts (Adderall) as a first line agent in the treatment of attention deficit hyperactivity disorder is evaluated. This report uses decision-tree analysis to evaluate the cost effectiveness of basing decisions on three treatment arms: initiation with methylphenidate, initiation with amphetamine/dextroamphetamine or no treatment. Data inputs such as efficacy rates, side effects, compliance rates and school administration rates were extracted from a literature review. A societal perspective was used to estimate outcomes in terms of incremental cost and incremental utilities over the time horizon of 1 year. PMID- 19807537 TI - Estimating the cost effectiveness of alternative drug treatments for postmenopausal osteoporosis. AB - The burden of postmenopausal osteoporosis is substantial and is likely to grow as the baby-boom generation ages. A greater understanding of the costs and clinical benefits of alternative therapies for postmenopausal osteoporosis is needed to develop informed health policies to reduce the future burden of osteoporosis. However, much of the data needed to evaluate the cost effectiveness of alternative therapies for postmenopausal osteoporosis are not available from clinical trials. Thus, cost-effectiveness models must cope with a high degree of uncertainty. This article provides a brief summary of the methodologic issues in cost-effectiveness analyses of current osteoporosis therapies, a review of the findings in published cost-effectiveness studies, and a discussion of challenges likely to be encountered in future economic evaluations of emerging osteoporosis therapies. PMID- 19807538 TI - Outcomes associated with inhaled corticosteroid use in asthma and COPD: an update. AB - The aim of this review is to report the updates in the medical literature on the outcomes associated with inhaled corticosteroid use. This update finds that inhaled corticosteroids remain a frontline choice as controller therapy in the treatment of asthma. Adherence to medications, especially inhaled corticosteroids used as controller medications, is responsible for a decrease in hospital, emergency room and physician visits. All of these translate to reduced asthma related healthcare costs and an increase in health-related quality of life. There is a need for further research on the role of inhaled corticosteroids in chronic obstructive pulmonary disease outcomes. PMID- 19807539 TI - Utility values for obesity and preliminary analysis of the Health Outcomes Data Repository. AB - Obesity is a global health problem, and comprehensive economic evaluations of treatments for obesity are needed to assess how it can be best addressed. Understanding the relationship between quality of life and obesity is important if the effects of treatment are to be fully captured by economic evaluations. All published estimates of utility related to obesity are reviewed. It was evident that there was widespread variability in the methods and findings. The authors' analysis of data on 13,000 patients within the Health Outcomes Data Repository shows a clearer picture. Obesity has a direct impact on health-related quality of life, therefore, economic evaluations need to incorporate these effects. PMID- 19807540 TI - Interferon-alpha in high-risk melanoma patients. AB - The incidence of melanoma has been climbing steadily since the early 1970s. Although melanoma can be successfully cured by surgical excision in its early stages, it is the most common fatal form of skin cancer. The most critical factors in determining the prognosis for patients with melanoma are primary tumor thickness, ulceration and the status of regional lymph nodes. Surgical treatment alone is inadequate in patients with thick, ulcerated tumors and those with nodal disease, as shown by poor 5-year disease-specific survival rates. Despite the fact that high-dose interferon-alpha has been approved for the treatment of high risk melanoma by the US Food and Drug Administration and regulatory agencies worldwide, current treatment recommendations vary widely due to the conflicting trial data and significant toxicity and cost associated with high-dose interferon alpha. PMID- 19807541 TI - Pharmacoeconomic analysis of therapies for pediatric patients. PMID- 19807543 TI - Cost-effectiveness of pegylated IFN-alpha2b and -2a and ribavirin for chronic hepatitis C treatment. AB - Pegylated interferon-alpha(2a) (Pegasys, Roche) and pegylated interferon alpha(2b) (Peg-Intron, Schering-Plough) with ribavirin (Copegus, Roche; Rebetol, Schering-Plough) is now acknowledged as the therapy of choice for chronic hepatitis C. This article reviews the existing evidence concerning the cost effectiveness of pegylated interferon and ribavirin versus nonpegylated interferon and ribavirin for the treatment of chronic hepatitis C and highlights key issues for future research. Recently published cost-effectiveness analyses suggest that use of pegylated interferon and ribavirin should reduce the cumulative incidence of liver complications, increase life expectancy, improve quality of life, and be cost-effective in comparison with nonpegylated interferon and ribavirin. The cost and benefits of the treatment depend on factors such as patient age, sex, viral load, histological severity and viral genotype. Studies of the natural history of chronic hepatitis C and randomized clinical trials of pegylated interferon and ribavirin versus other therapies need to be performed in special populations of patients to provide valuable data that could be incorporated into future cost-effectiveness analyses. The direct comparison of two marketed pegylated interferons in terms of cost-effectiveness is needed. The cost-effectiveness analyses of the two pegylated combination products should be preceded by a head-to-head clinical trial that would provide evidence of comparative efficacy. PMID- 19807544 TI - Idiopathic- and scleroderma-related pulmonary arterial hypertension: outcomes and QOL on bosentan. AB - Idiopathic pulmonary arterial hypertension (previously known as primary pulmonary hypertension) is a devastating disease of insidious onset, late diagnosis, progressive functional disability and poor prognosis. However, treatment with pulmonary arterial hypertension-specific agents has already changed the outlook wherever these agents are accessible. The process of economic evaluation of treatment for idiopathic pulmonary arterial hypertension is challenging due to limitations common to clinical studies in rare diseases, such as small patient numbers limiting survival information, lack of directly comparable trials for the efficacy of different drugs and a paucity of quality of life measurements. Bosentan (Tracleer), Actelion), has proven effective in clinical trials, and is the most frequently used pulmonary arterial hypertension-specific agent worldwide. It has additionally undergone a comprehensive quality of life trial and economic evaluation as therapy for idiopathic pulmonary arterial hypertension and pulmonary arterial hypertension associated with scleroderma. This review explores the role of bosentan in providing cost-effective therapy compared with alternative treatments, and addresses the future considerations required to ensure accessible patient care for idiopathic pulmonary arterial hypertension and pulmonary arterial hypertension associated with scleroderma. PMID- 19807545 TI - Half standard deviation estimate of the minimally important difference in HRQOL scores? AB - In addition to statistical significance, it is important to evaluate the magnitude of differences in health-related quality of life over time. Interest in establishing the minimal difference that is clinically important or the minimally important difference has burgeoned over the last few years. This review summarizes some of the leading approaches to estimating the minimally important difference, offers caveats on the minimally important difference estimation based on existing literature and provides recommendations for future work. The authors recommend using multiple anchors to estimate the minimally important difference, using only anchors that correspond to minimal change in health-related quality of life, reporting information about the variation around the estimates, and providing bounded estimates to reflect the uncertainty. PMID- 19807546 TI - Prostate cancer and QOL: impact of treatment, disease burden and psychosocial interventions. AB - Prostate cancer is highly prevalent and the second leading cause of cancer related deaths in American men. Treatment of localized prostate cancer is highly effective but can result in urinary, bowel and sexual dysfunction, undermining quality of life. Within 5 years of treatment, approximately a third of men will have a rising prostate-specific antigen level necessitating hormonal therapy which has a broad range of detrimental side effects. Moreover, the impact of hormonal therapy may overlay existing issues (e.g., comorbid illness) common to older men. Men with advanced disease experience significant disease- (e.g., metastasis) and treatment-related impairments, which may further compromise quality of life. A substantial amount of literature indicates that cancer patients benefit significantly from psychosocial interventions, and psychosocial factors may serve to buffer prostate cancer patients from some disease- and treatment-related decrements in quality of life. While very few randomized clinical trials have been conducted with prostate cancer patients, recent results suggest that such interventions improve coping skills, increase prostate cancer knowledge, reduce treatment-related disruption of daily activities, diminish bother associated with sexual problems and enhance global quality of life. In order to better understand who benefits most from psychosocial interventions, additional studies are needed to more fully assess the potential mediators and moderators of these intervention findings. PMID- 19807547 TI - Cost-effectiveness lessons from disease-modifying drugs in the treatment of multiple sclerosis. AB - In this article, an overview of lessons from cost-effectiveness analyses of disease-modifying drugs for the treatment of multiple sclerosis is provided. Special attention is paid to the uncertainty in reaching a conclusion from the economic evaluations. If recent debates over multiple sclerosis drugs are any indication, the public's understanding of why one conducts an economic evaluation may be at least as important as the cost-effectiveness estimate from the analysis. PMID- 19807548 TI - Cost-utility of inhaled corticosteroids in patients with moderate-to-severe asthma. AB - Different inhaled corticosteroids can be used to treat asthma but their relative efficacy on quality of life and relative economic impact are mostly unknown. A decision model compared the cost-utility of beclomethasone, beclomethasone extrafine, fluticasone and budesonide in adult patients with either moderate or severe persistent asthma. The patients' health state was described by the Asthma Symptom Utility Index. Patients' consumption of healthcare resources, according to the health state, was elicited by a Delphi Panel. Within 2 months, beclomethasone-extrafine prolonged quality-adjusted life by 0.5-2.3 days, as compared with the other inhaled corticosteroids, and reduced asthma-related per patient costs by euro12-67. PMID- 19807549 TI - Costs of multiple myeloma and associated skeletal-related events in The Netherlands. AB - Multiple myeloma causes significant skeletal morbidity (e.g., pain and pathologic fractures) resulting from osteolytic lesions, and the associated medical costs can be considerable. Therefore, the authors of this review conducted a retrospective cost analysis to quantify the medical costs associated with skeletal complications in patients treated for multiple myeloma in The Netherlands. In total, 20 patient records were investigated, and costs are given in euros. This analysis demonstrated that skeletal complications related to multiple myeloma contribute significantly to the cost of care, comprising approximately 17% of the total cost of treatment. This study provides a model for further investigation of the health-economic impact of skeletal complications in patients with malignant bone disease. PMID- 19807550 TI - Management and costs of severe psoriasis: the role of new biologics. AB - It is well accepted that psoriasis has a profound impact on the life of the patient. Moderate-to-severe psoriasis frequently requires phototherapy or aggressive systemic treatment; unfortunately these treatments have major limitations. New biologic treatments are now available that offer distinct safety advantages and possible efficacy advantages over traditional systemic treatments. These biologic drugs are also considerably more expensive than traditional treatments (up to approximately US$30,000 per patient per year). The treatment of psoriasis and other inflammatory skin diseases with biologics could place a significant burden on the healthcare delivery system. In addition to judicious use of biologics, it is the task of the dermatology community to encourage healthcare delivery systems to eliminate the impediments to safe, effective and lower cost treatments such as phototherapy. Appropriate patient selection for biologic therapies is paramount. The goal of therapy for patients with psoriasis is to alleviate suffering in a cost-effective manner for the patient and the healthcare system as a whole. Much has been written on the use of biologic therapies for psoriasis and in this review the authors offer their perspective on the place of biologics in the psoriasis armamentarium. PMID- 19807551 TI - The truly remarkable universality of half a standard deviation: confirmation through another look. AB - In this issue of Expert Review of Pharmacoeconomics and Outcomes Research, Farivar, Liu, and Hays present their findings in 'Another look at the half standard deviation estimate of the minimally important difference in health related quality of life scores (hereafter referred to as 'Another look') . These researchers have re-examined the May 2003 Medical Care article 'Interpretation of changes in health-related quality of life: the remarkable universality of half a standard deviation' (hereafter referred to as 'Remarkable') in the hope of supporting their hypothesis that the minimally important difference in health related quality of life measures is undoubtedly closer to 0.3 standard deviations than 0.5. Nonetheless, despite their extensive wranglings with the exclusion of many articles that we included in our review; the inclusion of articles that we did not include in our review; and the recalculation of effect sizes using the absolute value of the mean differences, in our opinion, the results of the 'Another look' article confirm the same findings in the 'Remarkable' paper. PMID- 19807552 TI - Bridging the gap between health economic analysis and the real world. PMID- 19807554 TI - Harmonizing international health-related quality of life research: ISOQOL's 11th Annual Scientific Meeting. PMID- 19807555 TI - 6th Australian conference on quality of life: understanding subjective well being. PMID- 19807556 TI - Nesiritide for heart failure: impact on costs and complications. AB - Decompensated heart failure accounts for approximately 1 million hospitalizations in the USA each year with an estimated cost of US$11,000 per hospitalization. Despite this prevalence and cost burden, relatively few therapies for decompensated heart failure have been developed over the past 30 years. Although once the mainstay of treatment of decompensated heart failure, the use of positive inotropic agents has fallen into disfavor. Although these agents improve hemodynamics and ejection fraction, there is evidence that the positive inotropes increase the risk of adverse clinical outcomes and mortality. Nesiritide is a naturetic peptide that produces balance vasodilation, inhibits sympathetic nervous system activity, and promotes diuresis and naturesis. At the time the drug received Food and Drug Administration approval for marketing in the USA, it had been shown to produce hemodynamic improvements to an extent greater than placebo or nitroglycerin. However, evidence of benefit in terms of clinical improvement and other outcomes was lacking. Recent trials have found that nesiritide reduces hospital length of stay (although not statistically significant in all trials) and healthcare resource utilization in patients admitted to hospital with decompensated heart failure. In a randomized, controlled trial, nesiritide given in the emergency room reduced hospital admissions for heart failure compared with placebo/usual care. Preliminary data from an outpatient intermittent infusion trial of nesiritide found that patients receiving nesiritide had fewer hospital admissions than patients randomized to standard care. There is currently little objective evidence that therapies used routinely in the management of patients with decompensated heart failure are associated with improved outcomes. Data with positive inotropic agents suggest that they do more harm than good. There is a growing body of evidence that nesiritide is associated with improvements in clinical outcomes in decompensated heart failure including fewer complications, less healthcare resource utilization, and lower costs when compared with standard therapy. Despite this evidence, larger, prospective trials demonstrating the impact of nesiritide on the costs and complications in decompensated heart failure are needed. PMID- 19807557 TI - PRODISQ: a modular questionnaire on productivity and disease for economic evaluation studies. AB - Productivity costs are often an important cost component in economic evaluations of healthcare programs. However, there still remains a low degree of consensus on the exact measurement and valuation of these productivity costs. The development and testing of valid measurement instruments in patient and worker populations may give researchers better tools to perform economic evaluations. This review briefly reports on the development, testing and validity of the modular PROductivity and DISease Questionnaire (PRODISQ) for the measurement (and valuation) of productivity costs. It was tested in several Dutch samples of patients and workers. The modular questionnaire covers all relevant aspects of the relationship between health and productivity including absence from work, compensation mechanisms that may reduce productivity loss, reduced productivity at work (efficiency losses) and productivity costs at the level of organizations. Using this questionnaire may facilitate the validity of estimates of productivity costs in economic evaluations of healthcare programs. PMID- 19807558 TI - Cost minimization of on-demand maintenance therapy with proton pump inhibitors in nonerosive gastroesophageal reflux disease. AB - On-demand proton pump inhibitor maintenance therapy is one of the suggested strategies for patients with nonerosive gastroesophageal reflux disease who achieve symptom remission after 4 weeks of continuous treatment. This review evaluates the difference in cost and quality of life of six on-demand proton pump inhibitor maintenance strategies in patients with nonerosive gastroesophageal reflux disease. This was performed by decision analysis and Markov modeling of costs (from the perspective of the healthcare service and society) and utility up to 12 months. Median utility scores associated with each proton pump inhibitor ranged from 0.731 to 0.745 quality-adjusted life years and were not statistically different. Annual expected cost, however, was statistically different among the different drugs and the following cost-minimization ranking was obtained for median costs to the healthcare service and society, respectively: rabeprazole (euro 181, euro 295), pantoprazole (euro 223, euro 341), lansoprazole (euro 249, euro 370), omeprazole 10 mg (euro 297, euro 412), esomeprazole (euro 295, euro 419) and omeprazole 20 mg (euro 405, euro 528). Unit cost of proton pump inhibitors was the major determinant of cost to the healthcare service, while the number of productivity days lost due to symptoms was the major determinant of cost to society. In Italy, on-demand use of rabeprazole for maintenance therapy of nonerosive gastroesophageal reflux disease incurs the least cost in comparison with the other proton pump inhibitors evaluated. PMID- 19807559 TI - Impact of antipsychotic medication on the cost of schizophrenia. AB - A systematic literature search regarding the factors influencing the costs of schizophrenia and of comparative studies on the impact of antipsychotic treatment on the costs of schizophrenia between 1990 and 2004 was conducted. Altogether, 11 studies on factors influencing the costs of schizophrenia and 39 pharmacoeconomic studies on the comparative effects of antipsychotic medications on costs of schizophrenia were identified. Considering the factors influencing the costs of schizophrenia, the results of the review revealed only little systematic knowledge due to a great variation of methodologic standards and variables included. With regard to the effects of antipsychotic treatment, most studies compared atypical with conventional antipsychotic compounds. Results of these studies are inconsistent, and due to differences in methodologic standards, it is not possible to draw an evidence-based conclusion on the economic effects of a particular type of antipsychotic treatment. Consensus on the main outcome indicators of antipsychotic treatment and common standards of outcome measurement as well as economic analysis methods are necessary to overcome the current lack of empirical evidence. PMID- 19807560 TI - Towards a healthier discount procedure. AB - Most national guidelines for pharmacoeconomic research prescribe discounting, mostly of money and health against the same rate. There is much debate on whether this is adequate. Two theoretical arguments, the consistency argument of Weinstein and Stason, and the paralyzing paradox of Keeler and Cretin, are mostly responsible for the current standards. However, more recently, several authors have indicated that the basis to claim the necessity of using similar discount rates is rather weak, both practically and theoretically. In terms of finding a new theoretical basis on which to base discount rates for money and, in particular, health, Van Hout has made an important suggestion arguing that the discount rate for health could be based on the expected growth in life expectancy and the diminishing marginal utility related to such additional health. Similarly, Gravelle and Smith argue that if the value of health grows over time, discount rates that are used for costs cannot directly be applied to effects, but should be adjusted downwards. PMID- 19807561 TI - Budget impact analysis: review of the state of the art. AB - A budget impact analysis for a new pharmaceutical product provides estimates of the likely impact of the new drug on a healthcare decision maker's short- and longer-term annual budgets. Budget impact analysis are an essential part of a comprehensive economic assessment of a new pharmaceutical product and are increasingly required, along with cost-effectiveness analyses, before national or local formulary approval or reimbursement. Standards for the development, content and presentation of the results of a budget impact analysis have recently been proposed but detailed guidance on alternative methods for estimating budget impacts are not yet available. Methods for performing these analyses require either static or dynamic analysis techniques depending on the type of health condition and health impact of the new drug. Several factors, that are not generally needed for cost-effectiveness analysis, should be part of a comprehensive budget impact analysis including the size of the treated population, second-order costs, market diffusion rates for the new drug, and off label use. A review of the recent literature indicates that there is only a limited number of published budget impact analyses and these vary greatly in the methods used. Instead of publication in peer-reviewed journals, budget impact analyses are more frequently presented directly to decision makers as interactive computer programs designed to compute the budget impact for specific health plans using plan-specific inputs. It is recommend that a comprehensive approach to budget impact estimation be adopted, with the results being presented from both a societal perspective as well as from more limited perspectives depending on the needs of the decision maker. PMID- 19807562 TI - Provider incentives and prescribing behavior in Europe. AB - European policy makers have increasingly moved from primarily focusing on supply side regulation of pharmaceutical costs to offering incentives for providers' prescribing behavior. The nature and implementation of these provider-focused policies are as diverse as the health systems in which they are enacted. Some policies have undergone significant alteration since their initial implementation as health system design has changed and experience has yielded greater knowledge of the types of policy that achieve results in practice. Evaluation of prescribing incentive policies in several European countries is constrained by a lack of comprehensive information systems tracking prescriptions. Analysis of policy experience suggests that policy ought to be formulated with clarity and transparency of objectives, also keeping in mind the ethical concerns associated with rewards and fines for prescribing levels when the professional aspirations of doctors include maintenance of quality care and some degree of autonomous decision making. PMID- 19807563 TI - Quality of life of children with attention deficit hyperactivity disorder. AB - To review the results of studies of quality of life in children with attention deficit hyperactivity disorder, three databases were searched and quality of life findings were reviewed. Comparisons were made with population norms for four studies that used the 50-item parent-reported Child Health Questionnaire (CHQ PF50). Effect sizes were computed to estimate the clinical importance of differences in quality of life. In total, ten publications were identified representing 1382 children with attention deficit hyperactivity disorder. Five different quality of life measures were used and compared with norms for the CHQ PF50. Children with attention deficit hyperactivity disorder had more problems in all psychosocial domains and family activities. Effect sizes for aspects of physical health domains were small in size. Pooled effect sizes for the CHQ-PF50 psychosocial domains and family activities were as follows: mental health = 0.55; self-esteem = -0.75; parental impact - time = -0.85; role emotional/behavioral = -1.22; behavior = -1.44; parental impact - emotions = 1.45 and family activities = -1.67. Children with attention deficit hyperactivity disorder have a poorer quality of life than children in the general population. Quality of life is an important outcome that is starting to receive attention in studies of children with attention deficit hyperactivity disorder. The use of tools such as the CHQ-PF50 shows how additional and useful information can be obtained that is relevant to a child with attention deficit hyperactivity disorder and their family. PMID- 19807564 TI - Costs of physician services in pharmacoeconomic analyses. PMID- 19807566 TI - Use of fluvastatin following percutaneous coronary intervention. AB - The use of percutaneous coronary intervention to remove occlusions from coronary arteries has increased substantially over recent years. Concurrent with the use of percutaneous coronary intervention, the use of lipid-lowering medications, such as statins, has increased. The Lescol Intervention and Prevention Study showed significant reductions with statins in postpercutaneous coronary intervention cardiac events. The cost- effectiveness of initiating statin use for all percutaneous coronary intervention patients is favorable, with significant health benefits (including improved survival) for relatively low additional costs (e.g., pound 3207 per quality-adjusted life year in the UK). Fluvastatin is the lowest cost statin currently available (less than the price of generic statins). Key economic issues include estimates of the current use of statins at the time of percutaneous coronary intervention and the likelihood of switching from more expensive statins to fluvastatin. PMID- 19807567 TI - In the long run, healthcare costs appear to be related to overweight and obesity at younger ages. AB - Overweight and obesity are associated with a higher risk of cardiovascular disease and noncardiovascular disease morbidity and mortality, and shorter life expectancy. Despite declines in prevalence of other key major cardiovascular disease risk factors, the prevalence of overweight and obesity has reached epidemic proportions and it continues to rise. This has implications for future Medicare expenditures. Recent findings indicate that irrespective of interim changes in weight, being overweight or obese in young adulthood and middle age is significantly and positively associated with higher average annual and cumulative cardiovascular disease-related and total Medicare charges in older age. However, the prevention and control of overweight and obesity has proven difficult due to overconsumption of calorie-dense foods, sedentary lifestyles, and insufficient and ineffective treatment options. Clinicians should emphasize weight management strategies for all patients and public health authorities should implement policies that promote maintenance and achievement of healthy weight through systemic environmental changes. Emphasis on the primary prevention of weight gain from early life onwards by adoption of healthy lifestyles, and prevention and control of excess weight at all ages are urgently required to contain and end the obesity epidemic and to reduce healthcare costs incurred by older people. PMID- 19807568 TI - Feasibility and linguistic validity of the Swedish version of the PDQ-39. AB - The Parkinson's Disease Questionnaire - 39 is the most widely used disease specific health status questionnaire for Parkinson's disease. Evaluations of various language versions have focused on psychometric properties, whereas documented evaluations regarding linguistic validity, respondent burden and questionnaire content are largely lacking. Work with the Swedish Parkinson's Disease Questionnaire - 39 has taken these aspects into account, which has resulted in a linguistically revised version which is currently being evaluated. Initial observations indicate that the revision rectified linguistic but not nonlinguistic problems, and that its feasibility in terms of respondent burden in more advanced disease can be challenged. There are also indications for the need for content improvements. These experiences are discussed along with their implications. PMID- 19807569 TI - Cost-effectiveness of intensity-modulated radiation therapy. AB - Technical advances have given medicine the opportunity to refine current treatment techniques to improve outcomes. Computed tomography, magnetic resonance imaging and high energy linear accelerators are but a few examples of technology translating into clinical practice. Intensity-modulated radiation therapy is a form of 3D conformal radiation that is being increasingly incorporated into the management of patients with prostate cancer. As with any new technology, the cost of intensity-modulated radiation therapy is considerably greater than standard therapy. Economic models can be useful to compare treatments when this comparison cannot be performed in a clinical trial. A Markov Model was used to compare the use of intensity-modulated radiation with 3D conformal radiation therapy in the treatment of a 70 year old man with a good- and intermediate-risk prostate cancer. Cost data for men with Medicare insurance and prostate cancer treated with intensity-modulated radiation therapy and 3D conformal radiation therapy was obtained from the billing department at the Fox Chase Cancer Center (PA, USA). Utilities were collected from men undergoing intensity-modulated radiation therapy and 3D conformal radiation therapy for prostate cancer. Intensity modulated radiation therapy was found to be cost effective in the treatment of a 70 year old man with prostate cancer with a incremental cost-effectiveness ratio of USD 16,182/quality-adjusted life year for men with intermediate-risk prostate cancer and USD 17,448/ quality-adjusted life year for men with good-risk prostate cancer. Sensitivity analysis found that a longer time horizon of the analysis and younger age at treatment favorably impact the cost-effectiveness ratio. PMID- 19807570 TI - Cost-effective approaches to influenza prevention and treatment. AB - Influenza, the seventh leading cause of death in the USA, accounts for 35,000 deaths and over 200,000 hospitalizations annually in that country alone. Recent advances in influenza vaccines, diagnosis and treatment have created numerous options for practicing clinicians, as well as economic opportunities for the makers of vaccines, rapid diagnostic tests and antiviral drugs. Since influenza like illness affects up to half of the population each year, selective use of expensive tests and treatments is essential to the practice of cost-effective medicine. Over the past 5 years, dozens of economic evaluations of influenza vaccination, rapid testing and antiviral therapy have been published, many of which are free of commercial bias. The existing literature, drawing practical lessons for clinical practice is reviewed, and new developments on the horizon are explored including vaccines, surveillance methods, antiviral agents and pandemic preparedness. PMID- 19807571 TI - Practical approach for using Medicare data to estimate costs for cost effectiveness analysis. AB - Many methods have been used to measure costs for cost-effectiveness analysis in healthcare. A central challenge in cost estimation is determining the direct cost of medical goods and services from a societal perspective. This review applies the methodology for calculating Medicare reimbursements for physician services, hospital services and medications as a means of estimating healthcare costs from a US societal perspective. This review provides the tools and information needed to calculate direct medical costs related to in- and outpatient services provided by physicians and hospitals, as well as drug costs using Medicare reimbursement data. The data used in calculating Medicare reimbursements was obtained from the Centers for Medicare and Medicaid Services website. Methods for estimating costs for a particular service in a specific location using Medicare and Medicaid Services are described and demonstrated. A method based on Medicare data that uses the unadjusted geographic practice cost index and standard hospital base rate to estimate healthcare costs that can be generalized to the US population is described and demonstrated. This review provides cost-effectiveness analysts with the tools needed to calculate healthcare service costs for economic research. It contains links to all websites needed for obtaining Medicare and Medicaid Services data and provides a step-by-step analysis of the methodology involved in calculating costs. A practical guide for applying the methodology used by Medicare and Medicaid Services to calculate direct medical costs in order to estimate US societal costs in cost-effectiveness analysis is provided. PMID- 19807572 TI - Review of the costs of illness of ankylosing spondylitis and methodologic notes. AB - This article reviews the cost of illness of ankylosing spondylitis in the literature and identifies limitations of comparability of cost of illness studies. The literature was searched semisystematically for studies that aimed to assess the cost of illness of ankylosing spondylitis from the societal perspective. Studies were appraised for methods and results following a self composed checklist. To compare the aggregated costs between the studies, adjustments for differential timing and purchasing power parities between the countries were applied. In total, 53 titles were retrieved by a MEDLINE search. Five articles reported on the costs of illness in four patient populations, one from the USA and three from Europe. All studies were prevalence studies with a bottom-up approach and reported direct and productivity costs. Patient characteristics differed with respect to sampling source, age, disease duration, presence of spondylitis-related comorbidity and employment status. Categories of resource use and costs were especially difficult to compare with regard to visits to types of healthcare providers and use of formal and informal help in relation to inability to perform unpaid work. In addition, not all studies reported the productivity costs based on human capital as well as the friction cost method. The cost per unit of resource use was only provided explicitly in one publication. Sensitivity analyses were usually not performed. Total 2002 costs based on the human capital approach varied between USD 7243 and 11,840, and productivity costs accounted for 53-73% of the total costs. Total 2002 costs based on the friction costs varied between USD 3353 and 3903, and productivity costs accounted for 15-26% of the total costs. Cost drivers of the direct costs varied among the studies. Physical functioning and/or disease activity were consistent determinants of total costs. In conclusion, between four bottom-up prevalence studies in ankylosing spondylitis from different countries, there were differences in characteristics of patients studied, inclusion of unpaid help and unpaid production loss in the costs and choice of human capital compared with the friction cost method to calculate the productivity costs. The absence of information of the unit-cost per resource hampers comparability. Overall, the ankylosing spondylitis-related costs of illness are substantial and the high costs of formal or informal help and work disability reflect the impact of the disease on physical functioning. PMID- 19807573 TI - Cost implications of new evidence on prevention of cardiovascular disease. AB - Due to the high morbidity, mortality and medical costs associated with cardiovascular diseases, the efficiency of western healthcare economies as a whole depends largely on the cost effectiveness of cardiovascular disease preventive interventions. In this review, the cost implications of new evidence relating to the treatment of cardiovascular disease risk factors are examined. In patients with hypertension, low-dose diuretics have been proven to be both the most effective and least costly first-line therapy. Recent data support more aggressive use of antihypertensive and lipid-lowering therapies in patients with myocardial infarction and other high-risk individuals such as diabetics. The use of behavioral and pharmacologic smoking cessation interventions and emerging screening or diagnostic approaches such as C-reactive protein testing and coronary calcium scanning are also discussed. PMID- 19807574 TI - Cost offset of lipid-lowering drugs for primary and secondary prevention of cardiovascular disease. AB - This review aims to perform an adherence-based pharmacoeconomic evaluation of patients receiving statin therapy. A population-based study was performed. All patients receiving statins between January 1996 and December 1999 were included in the study. Both prescriptions for statins and cardiovascular events recorded from enrolment to December 2000 were considered. A total of 18,596 patients were included in the study. The positive cost offset of treatment was demonstrated (euro 30.70 for cardiovascular events avoided per euro 100 allocated for statin therapy). The increasing of the percent cost offset was found to be related to older age, male gender and secondary prevention. This study underlined low rates of adherence to statin therapy, thus many patients may receive no or limited benefits from pharmacologic treatment. PMID- 19807575 TI - Socioeconomic disparities in health: the impact of pharmacotherapy. AB - Along with age and gender, socioeconomic status is a fundamental driver of health. This review discusses the meaning of socioeconomic status and how it influences health across a person's life. Particular attention is paid to the role of psychosocial stress and self-determination. Through various pathways, socioeconomic status may affect biologic aging. This review also discusses the implications of the socioeconomic status-health relationship for understanding international differences in population health such as differences in life expectancy. The review concludes with a discussion of the implications of these findings for national policies and the potential impact of pharmacotherapy. PMID- 19807577 TI - Measuring health-related quality of life in children: difficulties and challenges. PMID- 19807576 TI - Health insurance, access to prescription medicines and health outcomes in children. AB - Ensuring optimal access to medications has received increasing attention as healthcare systems struggle with increasing costs. Although this has been studied extensively in adults, there has been little investigation in pediatric populations, which have different healthcare needs. A literature review was conducted to examine the evidence regarding the relationship between insurance mediated access to prescription medicines and outcomes in children. In total, 12 studies were classified according to uninsured versus insured, type of insurance provider and impact of family income. The studies demonstrated that insurance coverage and low-cost sharing are both essential to facilitate access to medications. Increased access was consistently observed for insured compared with uninsured children. Access to prescription drugs frequently differed by type of health provider organization. Adequate family income was an important determinant of access to and receipt of prescriptions. Moreover, income-indexed insurance coverage may increase unmet need. Compared with the literature on access to prescription medicines and health outcomes in adults, there have been few studies in children. Further research relating pharmaceutical policies to pediatric health outcomes is needed to strengthen the quality of policy decision making regarding access to prescription medicines for children. PMID- 19807578 TI - Economic evaluation tailored to promote vaccine uptake: how third world consumers can respond. PMID- 19807580 TI - A pharmacoeconomic review of adalimumab in the treatment of rheumatoid arthritis. AB - The past 10 years has witnessed a major transformation in the treatment of rheumatoid arthritis, a chronic condition that leads to significant morbidity, impairment in quality of life and mortality. Adalimumab joins a class of biologic response modifiers that prevent joint destruction and maintain functional status. For expensive interventions such as biologic response modifiers to be a valuable use of healthcare resources, they must lower healthcare costs by reducing the prevalence of hospitalizations, assist people with rheumatoid arthritis in maintaining employment and improve patient quality of life. The rheumatoid arthritis market is competitive and growing quickly. Policy makers are faced with decisions surrounding the value of biologic response modifiers over conventional therapies, and whether one biologic response modifier has an advantage over another. PMID- 19807581 TI - Self-perceived health status of schizophrenic patients in Spain: analysis of geographic differences. AB - This report explores the use of regression models for estimating health status of schizophrenic patients from a Bayesian perspective. The aims are: to obtain a set of values of health states of the EQ-5D based on self-assessed health from a sample of schizophrenic patients; and to analyze the differences in the health status and in patients' perceptions of their health status between four mental health districts in Spain. The authors develop two linear models with dummy variables. The first model seeks to obtain an index of the health status of the patients using a visual analog scale as a dependent variable and the different dimensions of EQ-5D as regressors. The second model enables analysis of the differences between the self-assessed health status in the different geographic areas and also the differences between the patients' self-assessed health states, irrespective of their actual health state, in the different geographic areas. The analysis is done using a Bayesian approach with Gibbs sampling (computer program WinBUGS 1.4). Data concerning self-assessed EQ-5D with visual analog scale from four geographic areas of schizophrenic patients were obtained for the purposes of this analysis. The health status index for this sample was obtained and the differences for this index between the four geographic areas were analyzed. The study reveals variables that explain the differences in patients' health status and health state assessment. Four possible scenarios are considered. PMID- 19807582 TI - Pneumococcal polysaccharide vaccine: cost-effectiveness recommendations in adults. AB - Pneumococcal vaccination is an effective intervention to prevent pneumococcal infections. Earlier randomized clinical trials and most case-control studies have shown a significant protection against invasive pneumococcal diseases and pneumococcal pneumonia in the elderly and chronic patients with a higher risk of pneumococcal infections. However, more recent randomized clinical trials have only detected a significant protection in immunocompetent individuals. A review of the studies assessing the cost-effectiveness of the 23-valent polysaccharide vaccine has shown that pneumococcal vaccination should be recommended in all individuals aged over 64 years, and in chronic patients with a higher risk of pneumocccal infections. In these population groups, vaccination costs are lower than healthcare costs reduced with the intervention. In individuals aged over 64 years, cost savings could be obtained when the vaccine effectiveness is higher than 30%. Recent cost-effectiveness studies have shown that pneumococcal vaccination could also be recommended in individuals aged between 45 and 64 years without risk factors, since the cost-effectiveness ratio is lower than euro 30,000 per life-year gained. In individuals aged between 45 and 64 years, cost savings could be obtained when the vaccine effectiveness is higher than 90%, and when the percentage of pneumonias caused by Streptococcus pneumoniae are 40%. PMID- 19807583 TI - Economic and patient-reported outcomes of oral triptans in the treatment of migraine. AB - Migraine places a tremendous burden on patients, employers, the healthcare system and society. Triptans often effectively reduce pain and suffering caused by migraine and return patients to their typical routine. Seven triptans are marketed in the USA, and prescribers and payers need information to compare their effectiveness and value. However, studies that compare the treatment and economic impact between the seven triptans are limited. Meta-analyses are a useful source of information to make comparisons between the triptans and provide essential information for clinical and formulary decision makers. Future research should assess and expand upon the use of composite outcome measures that capture the desired response to treatment, consider the placebo effect in migraine, and make comparisons among the available triptans. PMID- 19807584 TI - Comparing generic preference-based health-related quality-of-life measures: advancing the research agenda. AB - Interest in generic preference-based health-related quality-of-life measures has grown considerably in recent years. Given the availability of several different measures, there is a question over the extent to which different measures produce different results. To determine the interchangeability between measures, a number of head-to-head comparisons have been undertaken to assess levels of agreement or association. However, the assessment of interchangeability may be addressed using a number of different methods. This paper reviews the methods that have been employed to examine the degree of interchangeability between the Assessment of Quality of Life, EuroQol-5D, Health Utilities Index Mark III, Short-Form-6D, Quality of Wellbeing and 15-dimension measures. It suggests a need to develop alternative econometric strategies and to explore, more fully, economic concepts of validity. PMID- 19807585 TI - Buprenorphine, buprenorphine/naloxone and methadone maintenance: a cost effectiveness analysis. AB - The use of heroin in Australia results in disproportionate harm. Although the evidence suggests that a relatively low proportion of the population aged 14 years and over have ever used (1.4%) heroin or have used it in the past 12 months (0.2%), heroin use remains a significant cause of death, injury, illness and social harm. Research demonstrates that being in treatment leads to less heroin use, lowered mortality rates and reduced crime. Pharmacotherapy treatment in Australia involves methadone and buprenorphine. Trial data used by Doran and colleagues are used in the current analysis to extend the original analysis of methadone versus low-dose buprenorphine to include high-dose buprenorphine and the buprenorphine-naloxone combination in the maintenance of heroin dependence. Adopting a provider perspective suggests that the observed difference between the cost-effectiveness of methadone and the other treatments was not statistically significant, indicating that high-dose buprenorphine and the buprenorphine/naloxone combination can provide a viable alternative to methadone in the treatment of heroin dependence. Wider treatment choices provide greater potential to recruit a larger proportion of regular dependent users and retain them in treatment for longer. The forthcoming introduction of buprenorphine/naloxone to Australia provides an exciting opportunity to enhance the treatment of heroin dependence in this country. PMID- 19807586 TI - Cost-effectiveness of rotavirus vaccines. AB - Rotaviruses are the commonest cause of severe watery diarrheal disease in infants and young children, causing an estimated 352,000-592,000 deaths per year, mostly in developing countries. In 1999, the first rotavirus vaccine to be licensed in the USA (RotaShield) was withdrawn . However, before use of the vaccine was suspended, several economic evaluations were performed. Recently, two rotavirus vaccines, Rotarix and RotaTeq, have completed Phase III clinical trials. The first economic evaluations of these new rotavirus vaccines are now appearing. The purpose of this paper is to review the existing cost-effectiveness evidence-base, and to provide methodologic suggestions for future analyses. PMID- 19807587 TI - Cost-effectiveness of HMG-CoA reductase inhibitors in the treatment of dyslipidemia and prevention of CHD. AB - The recent completion of several clinical trials of lipid-lowering therapy with 3 hydroxy-3-methylglutaryl coenzyme A inhibitors, or statins, was followed by an update in 2004 concerning the Adult Treatment Panel III guidelines issued by the National Cholesterol Education Program. Within this update, individuals considered at very high risk for coronary heart disease-related events were addressed, wherein the role of an aggressive, intensive lowering of lipid levels to new goals was presented. In achieving target cholesterol levels, the statins collectively represent first-line pharmacotherapeutic strategies, although each of the currently marketed agents differ in relative potency, dose, side effects and cost. As such, research concerning the pharmacoeconomics of lipid-lowering therapy may serve to augment clinical, evidence-based approaches to care. Furthermore, identifying and rectifying suboptimal care within healthcare systems may afford optimal outcomes amongst patients for resources consumed. PMID- 19807588 TI - Modeling risk of gastrointestinal events among Medicaid NSAID users using propensity scores. AB - The purpose of the study is to evaluate the gastrointestinal risk of nonsteroidal anti-inflammatory drugs compared with cyclooxygenase-2 inhibitors in a Medicaid managed care population. Medical and prescription claims were analyzed for all Medicaid-enrollees aged 18 years and older, who received a cyclooxygenase-2 or other prescription nonsteroidal anti-inflammatory drug between January 6, 2000 and January 6, 2002, and who did not use these drugs for at least 6 months prior. A logistic model was developed of the propensity for either treatment and stratified patients by quintiles of their propensity score, adjusting for demographics, indications for nonsteroidal anti-inflammatory drugs and gastrointestinal risks. The rates of gastrointestinal event (ICD-9 codes 531 534,578) among nonsteroidal anti-inflammatory drug and cyclooxygenase-2 users were compared. The model was adjusted for drug exposure which was calculated using the medication possession ratio. Of the total sample, 73% were female, 43% were Caucasian and 29% were older than 50 years. Both the direct and the propensity adjusted model, controlling for gastroprotective agents, and medication possession ratio showed significantly different rates of gastrointestinal events in nonsteroidal anti-inflammatory drug users as compared with cyclooxygenase-2 users (odds ratio = 1.874, 95% confidence interval 1.056, 3.326) and (odds ratio = 2.088, 95% confidence interval 1.061, 4.110) respectively. A significant difference in gastrointestinal event rates was found among patients in this Medicaid population on nonsteroidal anti-inflammatory drug versus cyclooxygenase-2 inhibitors, when the gastroprotective agent use and the medication possession ratio were controlled for. Misclassification bias was accounted for by adjusting for length of drug exposure. PMID- 19807589 TI - Quality of life as an endpoint in Phase I oncology clinical trials of novel chemotherapy drugs. AB - This paper examines the rationale, utility and feasibility of including quality of life as an outcome measure in Phase I trials of new applications of chemotherapy drugs. Typically, Phase I trials in oncology are designed to assess safety and maximal tolerated dose; however, it is argued that when subjectively assessed, self perceived quality of life is as important as physical toxicity. The outcomes of studies that have applied quality-of-life assessment in Phase I trials are reviewed, and recommendations are made for future research based on both methodologic and practical considerations. PMID- 19807590 TI - Depression and health-related quality of life in patients with rheumatoid arthritis. AB - Rheumatoid arthritis is a chronic inflammatory musculoskeletal disease with a prevalence rate of 1-3% in Western countries. In addition to limited physical function, rheumatoid arthritis patients also suffer from psychologic comorbidities. Depression in rheumatoid arthritis patients has been linked to disease outcomes such as increased healthcare service utilization and poor adherence to medication. This review focuses on the impact of depression on rheumatoid arthritis patient's quality of life and how quality of life can be improved through the management of depression. Both generic and disease-specific instruments have been used to assess health-related quality of life in rheumatoid arthritis research. Commonly used instruments include the generic Short Form-36, the Nottingham Health Profile, the disease-specific Arthritis Impact Measurement Scale and the Rheumatoid Arthritis Quality of Life. Studies have shown that depression in rheumatoid arthritis patients may exert an important impact on multiple domains of health-related quality of life. Currently, depression in rheumatoid arthritis has been managed by both psychoeducation and antidepressant treatment. They are generally effective in improving clinical outcomes. However, future studies are needed to clarify whether they can improve patient reported quality of life. As depression is a prevalent comorbidity in rheumatoid arthritis, clinicians should pay more attention to the rheumatoid arthritis patient's psychologic wellbeing. Screening for depression and other psychologic distress should be recognized as an important process in the routine clinical care of patients with rheumatoid arthritis. PMID- 19807591 TI - Developing benchmarks for adherence studies. PMID- 19807593 TI - Paclitaxel: cost-effectiveness in ovarian cancer. AB - Ovarian cancer accounts for a significant burden of healthcare costs worldwide. Therapy of this disease consists of a combined surgical and chemotherapeutic approach. Remarkable advances in chemotherapy have been made with the introduction of new agents such as paclitaxel. Based on positive clinical data from randomized trials, numerous cost studies have been undertaken to analyze the cost-effectiveness of paclitaxel. Reviewing all the available cost studies, the authors conclude that paclitaxel plus cisplatin treatment is cost effective. Paclitaxel demonstrated survival and utility gains in combination with cisplatin as first-line treatment in patients with Stage II-IV ovarian cancer compared with cyclophosphamide and cisplatin. Incremental costs of USD 6600-22,000 per life year gained are within an accepted range for new treatments. PMID- 19807594 TI - Healthy year equivalents versus quality-adjusted life years: the debate continues. AB - The quality-adjusted life year makes a number of restrictive assumptions about the nature of one's preference over health. The healthy year equivalent was originally proposed over 10 years ago as a major theoretic advance on the quality adjusted life year. This article reviews the theoretic, empiric and normative bases for the healthy year equivalent and quality-adjusted life year approaches to valuing health profiles that vary over time. Although critics have argued that the two-stage procedure proposed by the developers of healthy year equivalent is theoretically equivalent to a generalized time trade-off question, the notion of directly valuing a series of health states in a health profile has been an important contribution to the literature. Since then, new quality-adjusted life year approaches have been developed to combine health states that vary over time. However, these approaches have yet to be tested empirically. The empiric evidence comparing quality-adjusted life years with healthy year equivalents is equivocal, and the size and direction of difference is context specific. In some studies, no differences have been found. At the same time, where differences are observed, it is difficult to interpret the healthy year equivalent values as the gold standard since there is evidence of cognitive confusion in the answers being given to the valuations of complex health profiles. Based on the evidence to date, it is not clear that healthy year equivalents provide a better measure of preferences than quality-adjusted life years. PMID- 19807595 TI - Development of a questionnaire to assess the quality of care in Dutch dialysis centers from the patient's perspective. AB - This study aimed to develop and test a questionnaire in order to assess the quality of care in Dutch dialysis centers from the patient's perspective. The questionnaire is referred to as the Quality of Care in Dialysis centers Questionnaire. Focus group sessions were organized and the results were transformed into a 68-item test version of the Quality of Care in Dialysis centers Questionnaire. Factor analyses and item reduction were performed to construct the Quality of Care in Dialysis centers Questionnaire. The questionnaire has four dimensions: doctors, nurses, other staff members and facilities; with eight descriptive items plus one item to measure satisfaction per dimension. A visual analog scale was added to determine overall satisfaction. The Quality of Care in Dialysis centers Questionnaire is used in Dutch dialysis centers. Further research should be conducted to establish preference weights per dimension on the basis of the visual analog scale scores. PMID- 19807597 TI - Disease-related conditions in elderly patients with multiple sclerosis. AB - Improved therapies have increased life expectancy in multiple sclerosis patients, resulting in a group of patients aged 65 years and older. Nowadays, health professionals are caring for more and more elderly multiple sclerosis patients, but little is known about the specific problems of these individuals. A review including relevant papers on problems reported by elderly multiple sclerosis patients revealed that the number of studies addressing this topic is still very limited. Most of these studies included only small patient sample sizes (27-169 patients), and were not originally designed to investigate elderly multiple sclerosis patients. Information on the clinical course of multiple sclerosis in elderly patients revealed proportions of 62-90% with relapsing-remitting courses. Main problems reported included disabilities in mobility and activities of daily living. The findings on depressive symptoms and suicidal tendencies were inconsistent. Specific recommendations for treatment of multiple sclerosis in elderly patients were not found. The studies highlight the problem that health professionals should be aware of an increasing group of elderly multiple sclerosis patients with many specific needs, avoiding nursing home admissions being a particular challenge. Furthermore, studies on the problems of elderly multiple sclerosis patients are required. PMID- 19807596 TI - Health-related quality-of-life tools in heart failure. AB - Heart failure is one of the most debilitating chronic conditions that is likely to make ever-increasing demands on doctors' time over the next 10 years. Both the incidence and prevalence of the disease are predicted to rise substantially, defying the general downward trend reported in other cardiovascular disorders. Although momentous advances have been achieved in controlling the disease, patient-reported outcomes including health-related quality of life require more attention in patient-centered healthcare and clinical trials. There are many studies that use generic and condition-specific tools in heart failure patients. This special report provides a concise digest of condition-specific health related quality-of-life tools in heart failure from a clinical perspective. PMID- 19807598 TI - Socioeconomic relevance of selected treatment strategies in patients with chronic heart failure. AB - More than 2.8% of the population in the USA suffer from chronic heart failure, a condition that primarily afflicts people above the age of 60 years, and results in major expenses for social and health care of affected patients, their caregivers and families. The mainstay of treatment is drug therapy, complemented by comprehensive rehabilitation, invasive procedures, palliative treatment and monitoring. While sufficient health economic evidence exists on the cost effectiveness of certain standard drugs, such as angiotensin converting enzyme inhibitors or beta-blockers, little or no information exists on the cost effectivty of diuretics or aldosterone antagonists. Recently, introduced treatment strategies such as ventricular assistance devices or telemonitoring have yet to be evaluated for their cost-effectiveness in terms of risk of hospitalization, life expectancy and quality of life. With respect to the aging population and the drastically increasing costs of healthcare for heart failure patients, the goal is a more conscious and cost-effective resource allocation. This can help reduce the incidence of over and under treatment, diminish side effects and consequently improve symptoms, quality of life and life expectancy in the affected population. Further clinical trials and health economic analyses are necessary to optimize recommendations for healthcare of patients with chronic heart failure. PMID- 19807599 TI - Teachable moments: improving pediatric asthma outcomes through physician education. AB - In light of concerns about adherence to the 1997 National Heart Lung and Blood Institute guidelines as well as the 2002 update regarding treatment of asthma, the authors evaluate educational interventions designed to have a positive impact on pediatric patient outcomes. Social cognitive theory can provide the foundation of interventions for physicians that are intended to promote self efficacy in treating asthma patients. Emphasizing the need for physician education based on principles of adult learning, the authors call for more research in this area and encourage the incorporation of distance learning techniques into educational endeavors. The purpose of this article is to review research on the impact of healthcare provider education designed to increase use of practice guidelines on the quality of pediatric asthma care as reflected in patient outcomes and, ultimately, in cost of treatment. PMID- 19807600 TI - National trends in out-of-pocket prescription drug spending among elderly medicare beneficiaries. AB - This review examines the national trends in out-of-pocket and high economic burden of prescription drug expenditures by elderly people aged 65 years and older. From 1992 to 2000, prescription out-of-pocket drug expenditures increased at an annual rate of 7%. During this period the proportion of elderly without prescription drug coverage steadily decreased from 42% in 1992 to 21% in 2000. The proportion of elderly bearing high burden declined from 1992 to 1996 and steadily increased after 1997, despite the growth in prescription drug coverage. Taken together, these findings suggest that coverage expansion alone may not be sufficient to reduce the out-of-pocket prescription expenditures burden on the elderly. Some subgroups of the elderly, such as women and the chronically ill, were vulnerable to a high prescription expenditures burden throughout the 1990s. PMID- 19807601 TI - Improvement in health-related quality of life in patients with SLE following sustained reductions in anti-dsDNA antibodies. AB - Patients with systemic lupus erythematosus experience a decreased health-related quality of life due to disease activity, multisystem organ involvement and frequent hospitalization. High levels of anti-double-stranded DNA antibodies are associated with renal disease, a primary cause of morbidity and mortality in systemic lupus erythematosus, and progressive cognitive dysfunction. Post hoc analyses of two clinical trials of abetimus sodium identified responders with sustained reductions in anti-double-stranded DNA antibodies. At 6 and 12 months, responders reported improvement in health-related quality of life, as measured by the Medical Outcomes Survey Short Form 36, compared with no change or deterioration in nonresponders. Sustained reductions in anti-double-stranded DNA antibodies, regardless of treatment group, led to clinically meaningful improvements in patient-reported health-related quality of life. PMID- 19807602 TI - Economic aspects of epilepsy and antiepileptic treatment: a review of the literature. AB - The recent literature on the economic aspects of epilepsy and antiepileptic treatment are systematically reviewed. Studies for this literature review were selected by conducting a Medline literature search from January 1998 to October 2004. Studies reviewed had to follow one of the standard methods of health economics evaluation (cost of illness, cost-minimization analysis, cost effectiveness analysis, cost-benefit analysis and cost-utility analysis). A total of 31 epilepsy cost studies were reviewed. Cost-of-illness studies showed a marked difference in cost between countries and healthcare systems. Cost minimization analysis evaluations of four drugs with equivalent clinical efficacy found lamotrigine to be the most costly and carbamazepine the most economic. Cost effectiveness analysis studies found topiramate to be more cost effective than lamotrigine, and surgical lobectomy to be a very cost-effective treatment in the long term. Cost-benefit analysis studies generally focused on vagal nerve stimulation and epilepsy surgery, and found both treatment modalities to be significantly cost beneficial. Only two cost-utility analysis studies were performed and found long-term lamotrigine treatment to be less economically effective than most other pharmacologic treatments of serious disorders. Vagal nerve stimulation was found to be of questionable economic value and further research is necessary for clarification. The methodologic heterogeneity observed in the studies reviewed makes comparisons between them difficult. Nevertheless, many interesting interpretations arise from the results. Cost-effectiveness analysis studies were found to be much more credible than cost-minimization or cost-benefit analysis evaluations since they avoid efficacy of drugs being reduced to clinical efficacy parameters alone. Cost-utility analysis studies were found to be the most promising type of economic analysis since they are the only type of analysis that incorporates the patients' point of view. In conclusion, comparison across studies can only be achieved if future studies follow a common set of methods and similar economic-evaluation models. A collaborative effort of all experts involved is necessary if this is to be achieved. PMID- 19807603 TI - Review of cost-effectiveness studies of pegylated therapies for hepatitis C. AB - Hepatitis C virus is a worldwide public health problem and carries an increased economic burden that affects individuals, employers, insurers and health systems. Estimates suggest that by the year 2008, the yearly costs of hepatitis C virus will exceed US$1 billion. The authors conducted a literature search to identify studies on pharmacoeconomics and outcomes research for hepatitis C virus. According to the literature, the new standard for disease management is interferon-alpha, chemically linked with polyethylene glycol (pegylated interferon-alpha) as a monotherapy, or in combination with ribavirin. The authors reviewed a number of articles providing original research on outcomes related to pegylated therapies. Clinical outcomes and end points included sustained and early virologic response, disease progression, liver complications and mortality; economic outcomes included direct medical costs related to in- and outpatient costs, screening and diagnostics. Studies that consider health-related quality of life and fatigue were also identified. While most of the studies adopted Markov modeling to predict the likely outcomes and the effects of therapy, it is also important to consider indirect costs, such as productivity loss related to absenteeism. Future research on the use of pegylated interferons, weight-adjusted dosing of ribavirin, and the treatment of relapsers and nonresponders should provide valuable data that can be incorporated into cost-effectiveness studies. PMID- 19807604 TI - KIDSCREEN-52 quality-of-life measure for children and adolescents. AB - This study describes the development and reports the first psychometric results of the European KIDSCREEN-52 generic health-related quality-of-life questionnaire for children and adolescents. The KIDSCREEN-52, including ten dimensions, was applied in a European survey involving 12 countries (i.e., Austria, Switzerland, Czech Republic, Germany, Greece, Spain, France, Hungary, The Netherlands, Poland, Sweden and the UK) and 22,110 children and adolescents aged between 8 and 18 years of age. Questionnaire development included a literature search, expert consultation, and focus group discussions with children and adolescents. After definition of dimensions and collection of items, a translation process following international translation guidelines, cognitive interviews and a pilot test were performed. Analysis regarding psychometric properties showed Cronbach-alpha ranged from 0.77 to 0.89. Correlation coefficients between KINDL(R) and KIDSCREEN 52 dimensions were high for those assessing similar constructs (r = 0.51-0.68). All KIDSCREEN-52 dimensions showed a gradient according to socioeconomic status and most dimensions showed a gradient according to psychosomatic health complaints. The first results demonstrate that the KIDSCREEN-52 questionnaire is a promising cross-cultural measure of health-related quality-of-life assessment for children and adolescents in Europe. PMID- 19807605 TI - Are USA payers moving beyond pharmaceutical budget impact models and embracing cost-effectiveness analysis? PMID- 19807607 TI - Valuing a QALY: review of current controversies. PMID- 19807606 TI - Ethical economics and cost-effectiveness analysis: is it ethical to ignore opportunity costs? PMID- 19807609 TI - Risk 2005 conference. PMID- 19807610 TI - Person tradeoffs and the problem of risk. AB - Recently, the person tradeoff method has received much attention because it directs one to value health improvements of those other than one's self. As a result, several researchers have suggested that the person tradeoff measures preferences in pharmaceutical and health economic analysis more accurately than the standard gamble, which many consider the gold standard approach. However, the person tradeoff exercise as it is currently practiced assumes a riskless world where policy decisions lead to certain societal outcomes. In reality, health policy decisions are made under risk or uncertainty. Evidence suggests riskless and risky values differ for health improvements over persons. A problem with using the riskless person tradeoff method in economic evaluation of health is that the method conflates a change in the objects of measurement (i.e., a change from health gains with respect to self to gains with respect to others) with a change in the measurement context (i.e., a change from a risk context to a risk free one). Therefore, it is wise to examine what is sacrificed by ignoring risk and uncertainty inherent in health policy decision making. Given that the choices we make in the context of risk are often different than those we make under certainty, the authors argue that if one assumes context is important, the measurement of health improvements over persons should then be carried out in the context of risk. The authors propose the application of standard methods from utility theory to carry out elicitations for valuing health improvements of others within the context of risky choices. In addition, an alternative perspective of utility is discussed that in comparison puts forth a context free notion of utility. PMID- 19807611 TI - Medicaid service use and program costs for pregnant teens. AB - Teen pregnancy is an important public health issue for all teens, but particularly for low-income teens who rely on the public health safety net for services. Medicaid pays for more than two-thirds of deliveries among teenagers in the USA. To discern how this public program serves pregnant teens (aged 11-19 years), the authors used Medicaid enrollment and claims data for Florida, Georgia and New Jersey in 1995 to examine teens' enrollment duration, service use and average payments relative to 20-24-year-olds on Medicaid. Teens were more likely than the older women to have been enrolled in Medicaid before pregnancy and to have maintained coverage through the third month following delivery. If not enrolled prepregnancy, teens were more likely than older women to enroll later in pregnancy. Teens were less likely to receive early prenatal care and more likely to be hospitalized during pregnancy, usually for preterm labor. While total Medicaid payments for routine prenatal and delivery-related care were equivalent between teens and older women, payments for nonroutine care during pregnancy were modestly higher for teens in Florida and Georgia. Thus, only modest cost savings can accrue from lower average costs per pregnancy and delivery among teens who delay pregnancy. Additional and larger cost savings to the Medicaid program from preventing teen pregnancy would accrue from the expected lower enrollment in Medicaid among the teens as they age. PMID- 19807612 TI - Incorporating pharmacoeconomic and health outcomes into randomized clinical trials. AB - Clinical trials represent the most scientifically valid study design for evaluating the safety and efficacy of new medical treatments. However, for pharmacoeconomic outcomes, the usual efficacy clinical trial may not always provide the appropriate vehicle for generating information on the cost effectiveness of medical treatments. Pragmatic, or naturalistic, clinical trials may be more suited to understanding cost-effectiveness. There are several issues associated with designing clinical trials that include patient-reported outcomes or pharmacoeconomic outcomes. For patient-reported outcomes, it is important to match the domains and instruments with the objectives and characteristics of the target patient population, examine psychometric characteristics (i.e., reliability, validity and responsiveness) when selecting measures, and also consider practical issues related to data collection. Plans need to be put into place to minimize missing patient-reported outcome item level and form data. For pharmacoeconomic outcomes, it is necessary to collect healthcare resource data and to value these resources in some way. This may be challenging when multinational clinical trials are designed and conducted. There are challenges associated with the analysis of cost data and in conducting cost-effectiveness analyses. Design and reporting of prospective, pharmacoeconomic trials will help inform evidence-based medicine and hopefully improve the delivery and outcomes of medical care. PMID- 19807613 TI - The PedsQL as a pediatric patient-reported outcome: reliability and validity of the PedsQL Measurement Model in 25,000 children. AB - The PedsQL Measurement Model was designed as a modular approach to measuring pediatric health-related quality of life, and developed to integrate the relative merits of generic and disease-specific approaches. The PedsQL 4.0 Generic Core Scales have been translated into over 60 languages, with published data on over 25,000 children and adolescents in more than 75 peer-reviewed journals since 2001 for healthy children and numerous pediatric chronic health conditions. The PedsQL Disease and Condition-Specific Modules were designed to measure health-related quality-of-life dimensions specifically tailored for pediatric chronic health conditions, and include the PedsQL Asthma, Arthritis/Rheumatology, Cancer, Diabetes, Cardiac and Cerebral Palsy Modules, as well as the generic PedsQL Multidimensional Fatigue Scale, Pediatric Pain Questionnaire, Family Impact Module and Healthcare Satisfaction Module. The PedsQL has demonstrated reliability, validity, sensitivity and responsiveness for child self report for ages 5-18 years and parent proxy report for ages 2-18 years. The PedsQL has been shown to be related to other key constructs in pediatric healthcare such as access to needed care, healthcare barriers and quality of primary care. Future advances in the PedsQL() Measurement Model include web-based electronic administration (ePedsQL), integration into the electronic medical record, further efficacy and effectiveness outcome trials, including PedsQL ResourceConnect(SM) and PedsQL TIPS(SM), the development of the generic PedsQL Infant Scales for ages birth to 24 months and disease and condition-specific modules for other pediatric chronic health conditions. PMID- 19807614 TI - Economic evaluation databases as an aid to healthcare decision makers and researchers. AB - Economic evaluation databases have been developed to assist in setting priorities and facilitating research within the healthcare sector. This paper presents an overview of the major databases of economic evaluations currently available (HEED, NHS EED, the CEA Registry, CODECS, PEDE, EURONHEED and JEED). It describes the key features of each database and the main user groups. It also presents evidence of the value of access to economic evaluation databases, particularly for the researchers and decision makers who form their main target audience. The research available shows that both decision makers and researchers find economic evaluation databases helpful as a source of information. However, database producers may also need to better understand the requirements of their users and consider adaptations to their products. PMID- 19807615 TI - Treatment of insomnia: a review and update. AB - The purpose of this review is to provide an update on the treatment options available and their usage and outcomes in the treatment of insomnia in the USA. Both pharmacotherapy and behavioral therapy are recommended in the physician guidelines for insomnia management. Although pharmacotherapy can produce a rapid pharmacologic effect, for long-term effectiveness, behavioral therapy can be considered for chronic primary insomniac patients. The cost of behavioral therapy is a notable barrier to its prescription for patients with sleep difficulties. Increased utilization of both behavioral- and pharmacotherapy may reduce insomnia related healthcare costs and increase health-related quality of life. Further research should focus on the role of these therapies in outcomes of insomnia management. PMID- 19807616 TI - Towards optimal surgical outcomes. AB - Until relatively recently, quality in healthcare was difficult both to define and measure. Now that this is possible, healthcare providers must quickly adopt information technology to facilitate both the assessment of performance and improvement. Such improvements require recognition of the role of systems of care and the need to change these systems in order to improve performance. In the coming years, the tension between the pressure for quality improvement and the pressure for cost-containment is likely to increase. PMID- 19807617 TI - Adding clopidogrel to aspirin improves outcome in ST-elevation myocardial infarction patients receiving fibrinolytic therapy. AB - Acute coronary syndromes result from the rupture of an atherosclerotic plaque with superimposed thrombosis. In an ST-elevation myocardial infarction, the thrombus occludes the coronary vessel, leading to an abrupt decrease in myocardial perfusion. The focus of initial management is the timely restoration of flow in the infarct-related artery via fibrinolytic therapy or percutaneous coronary intervention. Adjunctive therapy aimed at inhibition of platelets and the coagulation cascade is critical to establish and maintain vessel patency. Clopidogrel, an oral antiplatelet agent, has recently been shown to offer significant clinical benefit in STEMI (ST-elevation myocardial infarction) and is a welcome addition to standard fibrinolytic therapy. PMID- 19807618 TI - Health-related quality of life and symptom research in glioblastoma multiforme patients. AB - Health-related quality of life is increasingly becoming an important end point in cancer research. Until recently, relatively little was known about the health related quality of life of glioblastoma multiforme patients. This is due to the relative scarcity of this disease compared with other cancers and its dismal prognosis. Now, there is progress in the treatment of this tumor: new (combination) treatments will be studied in randomized trials and health-related quality will be incorporated as a (secondary) end point next to progression-free survival. Glioblastoma multiforme patient health-related quality of life may also have prognostic significance and may, therefore, be used with the individual patient to tailor treatment. This review aims to chart the history of glioblastoma multiforme and treatment, reporting on the latest health-related quality of life tools and research findings. PMID- 19807619 TI - The market for antituberculosis drugs and vaccines: incentives for investment in new products. AB - The market for antituberculosis drugs is reviewed briefly and an estimate is formed of the potential US market for a new and improved tuberculosis vaccine. By 2010, global sales of all antituberculosis drugs are expected to reach USD 612 670 million annually. An urgent need for better antituberculosis drugs exists that could permit a shorter course and less frequent dosing so that tuberculosis treatment regimens could scale up more rapidly. Should a more effective tuberculosis vaccine become available, there are approximately 18 million high risk individuals in the USA alone for whom the vaccine would be demonstrably cost beneficial. The high-risk groups include healthcare workers, military personnel, HIV-infected individuals, migrant farm workers and prisoners. PMID- 19807620 TI - New management of the pregnant patient to reduce the risk of transmitting herpes to the newborn. AB - Growing understanding of the mechanisms and epidemiology of neonatal herpes simplex virus acquisition allows for the development of a new management of the pregnant patient, one that will reduce the risk of transmitting herpes to the newborn. Maternal herpes simplex virus infection, especially acquired during pregnancy, can result in transmission to the newborn. Infection of the CNS of the newborn results in death or significant neurodevelopmental impairment in most of these infected babies. Herpes simplex virus infection acquired prior to or during pregnancy has a physical and psychologic impact on the health of the mother. Newer, more sensitive and specific serology testing for herpes simplex virus in combination with antiviral pharmacotherapy is now recognized as an important option for reducing the risk of herpes simplex virus transmission from mother to newborn. This review discusses new information concerning the prevention, diagnosis and treatment options for the management of herpes simplex virus infection during pregnancy with a focus on the evolving role of prevention. PMID- 19807621 TI - Optimizing treatment policies and improving care: impact on outcome in patients with cystic fibrosis. AB - There is no cure for cystic fibrosis but recent advances in care have increased the average life expectancy to over 30 years. However, patients may find themselves prescribed over seven to eight different medications a day, some of which are laborious and time consuming to administer. The physician should balance potential benefits from treatments against quality-of-life requirements. In this review, the authors examine the place in the overall treatment regimen of recently available, and often expensive, drugs. The review concentrates on eradication regimens for early or recurrent Pseudomonas aeruginosa infection; inhaled tobramycin therapy; regular versus on-demand intravenous antibiotics; treatment of respiratory methicillin-resistant Staphylococcus aureus infection; the role of macrolide antibiotics and the role of inhaled dornase alfa in early treatment. PMID- 19807622 TI - Staphylococcus aureus bacteraemias: time to act. AB - Mandatory reporting and public sharing of information would likely lead to improved health care practices and save lives. PMID- 19807623 TI - Systemic sclerosis: new hope for an unyielding disease. AB - There is still no cure, but what advances have been made in managing this disabling condition? PMID- 19807624 TI - Reforming Australian health care. AB - Time to debate the recommendations of the National Health and Hospitals Reform Commission is slipping away. PMID- 19807626 TI - Bringing patients' own medications into an emergency department by ambulance: effect on prescribing accuracy when these patients are admitted to hospital. AB - OBJECTIVE: To determine whether the availability of patients' own medications (POM) in emergency departments (EDs) results in decreased prescribing errors of patients' usual medications on admission. DESIGN, PARTICIPANTS AND SETTING: Observational study of patients presenting by ambulance to the ED of Austin Hospital, a Melbourne metropolitan teaching hospital, between 13 and 31 March 2006. Patients were enrolled if they were brought to the ED by ambulance, aged 18 years or older, taking four or more regular medications, admitted to hospital, and not referred to a pharmacist before the admission medication chart was written. ED pharmacists determined patients' regular medications and details of medications brought in by ambulance. Admission medication charts were assessed and discrepancies were recorded as prescribing errors if a change was made after a pharmacist discussed the discrepancy with the prescriber. MAIN OUTCOME MEASURES: Percentage of medications correctly prescribed when POM were brought in to the ED compared with when they were not; the nature and frequency of prescribing errors on admission. RESULTS: 100 patients were enrolled; they were taking 4-17 regular medications (mean, 8.0; SD, 3.7). Among the 428 POM that were brought to the ED, 56 errors occurred (13.1%); and among the 372 regular medications taken by patients for whom POM were not brought in, 95 errors occurred (25.5%) (difference in percentages, 12.4%; 95% CI, 6.7%-18.0%; P<0.001). The most prevalent prescribing errors were omissions (40.4%), and most errors (72.8%) were classified as of "moderate" clinical significance. CONCLUSIONS: When POM were brought to the ED by paramedics, significantly fewer errors occurred on admission medication charts. An intervention program to encourage paramedics to bring POM to the ED is indicated. PMID- 19807627 TI - Initial impact of Australia's National Bowel Cancer Screening Program. AB - OBJECTIVE: To examine the initial impact of the National Bowel Cancer Screening Program (NBCSP), which was launched in May 2006 and offers faecal occult blood testing to Australians aged 55 or 65 years. DESIGN AND SETTING: Review of data on colorectal cancer (CRC) cases diagnosed between May 2006 and June 2008 from a prospective database used at 19 Australian hospitals, linked and analysed by BioGrid Australia. MAIN OUTCOME MEASURES: Number of CRC cases detected through the NBCSP or symptomatic presentation, and differences by sex, stage at diagnosis, tumour location and level of socioeconomic disadvantage. RESULTS: 1628 cases of CRC were identified; 1268 had information on the patients' test status as part of the NBCSP, and 40 of these (3.2%) were recorded as being detected by the NBCSP. Of 75 CRC cases in patients aged 55 or 65 at diagnosis, 22 were NBCSP detected. Overall, there was no difference in NBCSP-detected cases by sex. The distribution of tumour locations was similar between NBCSP-detected cases and symptomatic cases, but NBCSP-detected cancers were diagnosed at an earlier stage than symptomatic cancers (stage I, 40% v 14%; stage IV, 3% v 15%, respectively). Of patients diagnosed through the NBCSP, 63% were from areas of least socioeconomic disadvantage (deciles 8-10) and 18% were from the most disadvantaged areas (deciles 1-4) (P=0.0375). CONCLUSION: Initiation of the Australian NBCSP has had a measurable impact on CRC stage at diagnosis, and an improvement in survival would be anticipated. The lower uptake among people from disadvantaged areas is of concern. PMID- 19807628 TI - National health reform: it's time for a decision. AB - A national system financed and governed by the Australian Government would open up access to the widest range of health services. PMID- 19807625 TI - Staphylococcus aureus bacteraemia: a major cause of mortality in Australia and New Zealand. AB - OBJECTIVE: To document the types of, and mortality from, Staphylococcus aureus bacteraemia in Australia and New Zealand, and determine factors associated with mortality. DESIGN AND SETTING: Prospective observational study in 27 independent or hospital pathology laboratories in Australia (24) and New Zealand (3), employing a web-based database to prospectively record demographic features, selected risk factors, principal antibiotic treatment and mortality data on all patients with positive blood cultures for S. aureus from June 2007 to May 2008. MAIN OUTCOME MEASURE: 30-day all-cause mortality. RESULTS: 1994 episodes of S. aureus bacteraemia were identified, and complete 30-day follow-up data were available for 1865. Most episodes had their onset in the community (60.8%; 95% CI, 58.7%-63.0%). Methicillin-resistant S. aureus (MRSA) caused 450 episodes (24.1%; 95% CI, 22.2%-25.9%), and 123 of these (27.3%) had a susceptibility profile consistent with community-associated MRSA. All-cause mortality at 30 days was 20.6% (95% CI, 18.8%-22.5%). On univariate analysis, increased mortality was significantly associated with older age, European ethnicity, MRSA infection, infections not originating from a medical device, sepsis syndrome, pneumonia/empyema, and treatment with a glycopeptide or other non-beta-lactam antibiotic. On multivariable analysis, independent predictors of mortality were age, sepsis syndrome, pneumonia/empyema, device-associated infection with a secondary focus, left-sided endocarditis, and treatment with a glycopeptide such as vancomycin, but not MRSA infection. CONCLUSIONS: S. aureus bacteraemia is a common infection in both the community and hospitals in Australia and New Zealand, and is associated with appreciable mortality. Invasive MRSA infection may be more life-threatening, partly because of the inferior efficacy of the standard treatment, vancomycin. National web-based surveillance of S. aureus bacteraemia and its outcomes is not only important but also easily achievable. PMID- 19807629 TI - A healthier future for all Australians: an overview of the final report of the National Health and Hospitals Reform Commission. AB - After extensive community and health industry consultation, the final report of the National Health and Hospitals Reform Commission, A healthier future for all Australians, was presented to the Australian Government on 30 June 2009. The reform agenda aims to tackle major access and equity issues that affect health outcomes for people now; redesign our health system so that it is better positioned to respond to emerging challenges; and create an agile, responsive and self-improving health system for long-term sustainability. The 123 recommendations are grouped in four themes: Taking responsibility: supporting greater individual and collective action to build good health and wellbeing. Connecting care: delivering comprehensive care for people over their lifetime, by strengthening primary health care, reshaping hospitals, improving subacute care, and opening up greater consumer choice and competition in aged care services. Facing inequities: taking action to tackle the causes and impact of health inequities, focusing on Aboriginal and Torres Strait Islander people, people in rural and remote areas, and access to mental health and dental services. Driving quality performance: having leadership and systems to achieve the best use of people, resources and knowledge, including "one health system" with national leadership and local delivery, revised funding arrangements, and changes to health workforce education, training and practice. PMID- 19807630 TI - Final report of the National Health and Hospitals Reform Commission: will we get the health care governance reform we need? AB - The National Health and Hospitals Reform Commission (NHHRC) has recommended that Australia develop a "single health system", governed by the federal government. Steps to achieving this include: a "Healthy Australia Accord" to agree on the reform framework; the progressive takeover of funding of public hospitals by the federal government; and the possible implementation of a consumer-choice health funding model, called "Medicare Select". These proposals face significant implementation issues, and the final solution needs to deal with both financial and political sustainability. If the federal and state governments cannot agree on a reform plan, the Prime Minister may need to go to the electorate for a mandate, which may be shaped by other economic issues such as tax reform and intergenerational challenges. PMID- 19807631 TI - Staphylococcus aureus bacteraemia as a quality indicator for hospital infection control. AB - OBJECTIVE: To evaluate the practicality and effectiveness of a new program that made health care-associated Staphylococcus aureus bacteraemia (SAB) a quality indicator at Austin Health. DESIGN AND SETTING: Roll-out of the program over 9 months and review over 27 months from January 2006. Every episode of SAB at Austin Health was promptly reviewed, and classified as community- or health care associated and as inpatient- or non-inpatient-related. Feedback was provided to treating clinicians for every SAB episode considered potentially preventable, and education-based interventions were introduced where appropriate. MAIN OUTCOME MEASURE: Episodes of SAB associated with health care at Austin Health per 1000 separations (hospital discharges) per month. RESULTS: We identified 131 episodes of health care-associated SAB, of which 90 (68.7%) were caused by methicillin susceptible S. aureus, 96 (73.3%) occurred in inpatients, and 65 (49.6%) were associated with a vascular access device. The health care-associated SAB rate was 1.1 per 1000 separations in the first 9 months, and fell by 55% to 0.51 per 1000 separations in the subsequent 18 months. We estimated that there were 80 fewer SAB episodes (95% CI, 20-140) than expected had the initial rate remained unchanged, a national saving of $1.75 million to Austin Health over 27 months. About 16 hours per month of clinical nurse consultant time was required to maintain the program, representing a 0.1 equivalent full-time position, or a cost of $7000-$9000 per year. CONCLUSION: Introducing a structured program to investigate all health care-associated SABs, rather than only infections with methicillin-resistant S. aureus, revealed a large under-recognised burden of potentially preventable infections. The program was simple and low-cost, and the rate of health care-associated SAB has fallen significantly since its introduction. PMID- 19807632 TI - General Practice Super Clinics--how will they meet their educational objectives? AB - The Australian Government will provide $275 million over 4 years to general practice infrastructure across Australia with the rollout of 31 General Practice Super Clinics. One of the core objectives of these Super Clinics is to support medical education. Several studies have demonstrated that the major barriers to teaching in general practice are time, space and money. We argue that General Practice Super Clinics can provide a responsive, flexible work culture; and improved payment and targeted resources to support the need for increased teaching capacity, and to attract and retain workforce for general practice and primary care. PMID- 19807633 TI - An unusual case of recurrent fever, jaundice and right upper quadrant pain. PMID- 19807634 TI - A prolonged mumps outbreak among highly vaccinated Aboriginal people in the Kimberley region of Western Australia. AB - OBJECTIVE: To describe a prolonged outbreak of mumps in the Kimberley region of Western Australia in 2007-2008. DESIGN: Descriptive analysis of all mumps cases notified to the WA Notifiable Infectious Diseases Database for the period 1 July 2007 to 30 June 2008. MAIN OUTCOME MEASURES: Notified cases of mumps by patients' place of residence, age, Indigenous or non-Indigenous ethnicity, vaccination status and method of diagnosis. RESULTS: 84% (153/183) of mumps notifications in WA over the study period occurred in the Kimberley region or were directly linked to Kimberley cases. Median age of patients was 18 years (range, 2-63 years), and 54% of patients were aged less than 20 years. Almost all (92%) were Australian Aboriginal people; 67% (102/153) had received at least one dose of mumps vaccine, and 52% had received two doses. The highest notification rate (1816 cases per 100,000 population) was in the Aboriginal 15-19-years age group, and 92% of these patients had received at least one dose of mumps vaccine. Almost all outbreak cases (94%) were laboratory confirmed. Genotyping was performed on 20 mumps virus isolates: all were genotype J. CONCLUSION: A prolonged outbreak of mumps occurred in a well defined, highly vaccinated, predominantly young Aboriginal population in the remote Kimberley region of WA. This outbreak raises questions about the effectiveness and scheduling of the current vaccine (which is genotype A derived), especially for Aboriginal people. Surveillance of circulating mumps virus genotypes and neutralisation studies will help in evaluating the protection provided by the current vaccine against genotypically different strains. PMID- 19807635 TI - My story: balancing family, work and community. PMID- 19807636 TI - Inadequate data collection prevents health planning for released prisoners. PMID- 19807637 TI - Is viral nucleic acid testing of eye donors cost-effective? PMID- 19807638 TI - Cruelty towards the family pet: a survey of women experiencing domestic violence on the Central Coast, New South Wales. PMID- 19807639 TI - Effect of swimming pools on antibiotic use and clinic attendance for infections in two Aboriginal communities in Western Australia. PMID- 19807640 TI - The private hospital: a potential surgical training ground. PMID- 19807641 TI - Avoidable hospitalisation in Aboriginal and non-Aboriginal people in the Northern Territory. PMID- 19807642 TI - Are we ready for the next big thing? PMID- 19807643 TI - Prolonged varicella viraemia and streptococcal toxic shock syndrome following varicella vaccination of a health care worker. PMID- 19807644 TI - Making cars and making health care: a critical review. PMID- 19807645 TI - Non-invasive prenatal diagnosis--toward a new horizon. PMID- 19807646 TI - Smoking and The Simpsons. PMID- 19807647 TI - Chromobacterium violaceum endocarditis and hepatic abscesses treated successfully with meropenem and ciprofloxacin. PMID- 19807649 TI - Progress and prospects of chitosan and its derivatives as non-viral gene vectors in gene therapy. AB - Although cell transfection by viral vectors is highly efficient, undesirable side effects including immunogenicity, toxicity and carcinogenesis have to be taken into consideration before their clinical applications. In contrast, most non viral vectors, such as chitosan, are advantageous due to their biocompatibility, biodegradability, low toxicity and immunogenicity. However, the tranfection efficiency of chitosan as gene vector is rather low because of its low stability and low buffering capacity. Recent technological progress in chemical modification of chitosan has led to improvements of its transfection efficiency without disturbing its biocompatibility and biodegradability. These advances have led to a better understanding of the relationship between the physicochemical characteristics of a non-viral vector and its transfection efficiency. In this review, we summarize the obstacles encountered during the transfection process of chitosan and its derivatives, and then focus on strategies to overcome these obstacles. An accurate method for determining the rate-limiting step and intracellular unpacking kinetics of chitosan and its derivatives is also presented. Lastly, gene-silencing chitosan/small interfering RNA (chitosan/siRNA) complexes and prospects of feasible methods for enhancing the transfection efficiency of chitosan and its derivatives are discussed. PMID- 19807648 TI - Immunomodulatory gene therapy in lysosomal storage disorders. AB - Significant advances in therapy for lysosomal storage disorders have occurred with an accelerating pace over the past decade. Although enzyme replacement therapy has improved the outcome of lysosomal storage disorders, antibody responses have occurred and sometimes prevented efficacy, especially in cross reacting immune material negative patients with Pompe disease. Preclinical gene therapy experiments have revealed the relevance of immune responses to long-term efficacy. The choice of regulatory cassette played a critical role in evading humoral and cellular immune responses to gene therapy in knockout mouse models, at least in adult animals. Liver-specific regulatory cassettes prevented antibody formation and enhanced the efficacy of gene therapy. Regulatory T cells prevented transgene directed immune responses, as shown by adoptive transfer of antigen specific immune tolerance to enzyme therapy. Immunomodulatory gene therapy with a very low vector dose could enhance the efficacy of enzyme therapy in Pompe disease and other lysosomal storage disorders. PMID- 19807650 TI - Current advances in gene therapy for the treatment of genodermatoses. AB - Gene therapy provides the possibility of long term treatment for the severest of congenital disorders. In this review we will examine the recent advances in gene therapy for genodermatoses. Congenital diseases of the skin exhibit a wide range of severity and underlying causes and there are many possible therapeutic avenues. Gene therapy approaches can follow three paths-in vivo, ex vivo and fetal gene therapy, though the later is currently theoretical only it can provide potential results for even the most severe congenital diseases. All approaches utilize the many different vector systems available, including viral and the emerging use of non- viral integrating vectors. In addition, the use of RNAi based techniques to prevent dominant mutant protein expression has been explored as a therapy for specific dominant disorders such as keratin mutation disorders. Progress has been rapid in the past few years with some initial successful clinical trials reported. However, there are still some issues surrounding long term expression, transgene sustainability and safety issues that need to be addressed to further shift from experimental to clinically therapeutic applications. With the continuing development, merger and refinement of existing techniques there is an ever increasing likelihood of gene therapies becoming available for the more severe genodermatoses within the next decade or shortly thereafter. PMID- 19807651 TI - Cortical and putamen age-related changes in the microvessel density and astrocyte deficiency in spontaneously hypertensive and stroke-prone spontaneously hypertensive rats. AB - Cerebral small vessel disease (SVD) is a major contributor to dementia in the elderly, and hypertension represents a major cause for developing the disease. However, little is known about its development and progression. Modifications of large cerebral arteries due hypertension are thought to participate to the development of small ischemic infarcts, but the status of the small vessels before the establishment of hypertension is not well defined. Using spontaneously hypertensive rats (SHR) and stroke-prone SHR (SP-SHR) as a models for SVD, we analysed the effect of hypertension on the microvasculature in the cortex and putamen, and on its relationship with astrocytes in animals aged 2 to 9 months. Compared with the normotensive Wistar-Kyoto rats (WKY), the densities of the collagen type IV-positive capillaries were significantly higher in both brain areas of young SHR and SP-SHR. In contrast, the expression of the astrocytic marker GFAP was significantly lower in these animals, whereas astrogliosis was observed after 6 months in their cortex only. To investigate if chronic hypoxia occurs due to the lower number of astrocytes in young SHR and SP-SHR, we evaluated the levels of HIF-1alpha in both brain regions. The accumulation of HIF 1alpha was not observed at the youngest ages, but was apparent in neurons of 9 month-old SHR and SP-SHR. Our results indicate that the brains of young SHR and SP-SHR rats show evidence of cellular imbalance between microvessels and astrocytes at the neurovascular unit that may lead to their higher vulnerability to hypoxic events at older ages. PMID- 19807653 TI - Chronic methylphenidate-effects over circadian cycle of young and adult rats submitted to open-field and object recognition tests. AB - In this study age-, circadian rhythm- and methylphenidate administration- effect on open field habituation and object recognition were analyzed. Young and adult male Wistar rats were treated with saline or methylphenidate 2.0 mg/kg for 28 days. Experiments were performed during the light and the dark cycle. Locomotor activity was significantly altered by circadian cycle and methylphenidate treatment during the training session and by drug treatment during the testing session. Exploratory activity was significantly modulated by age during the training session and by age and drug treatment during the testing session. Object recognition memory was altered by cycle at the training session; by age 1.5 h later and by cycle and age 24 h after the training session. These results show that methylphenidate treatment was the major modulator factor on open-field test while cycle and age had an important effect on object recognition experiment. PMID- 19807652 TI - Preservation of cellular glutathione status and mitochondrial membrane potential by N-acetylcysteine and insulin sensitizers prevent carbonyl stress-induced human brain endothelial cell apoptosis. AB - Oxidative stress-induced cerebral endothelial cell dysfunction is associated with cerebral microvascular complication of primary diabetic encephaolopathy, a neurodegenerative disorder of long-standing diabetes, but the injury mechanisms are poorly understood. This study sought to determine the contribution of carbonyl (methylglyoxal, MG) stress to human brain endothelial cell (IHEC) apoptosis, the relationship to cellular redox status and mitochondrial membrane potential, and the protection by thiol antioxidant and insulin sensitizers. MG exposure induced IHEC apoptosis in association with perturbed cellular glutathione (GSH) redox status, decreased mitochondrial membrane potential (Deltapsi(m)), activation of caspase-9 and -3, and cleavage of polyADP-ribose polymerase. Insulin sensitizers such as biguanides or AMP-activated protein kinase activator, but not glitazones, afforded cytoprotection through preventing (Deltapsi(m) collapse and activation of caspase-9 that was independent of cellular GSH. Similarly, cyclosporine A prevented Deltapsi(m) collapse, while N acetylcysteine (NAC) mediated the recovery of cellular GSH redox balance that secondarily preserved Deltapsi(m). Collectively, these results provide mechanistic insights into the role of GSH redox status and mitochondrial potential in carbonyl stress-induced apoptosis of brain endothelial cells, with implications for cerebral microvascular complications associated with primary diabetic encephalopathy. The findings that thiol antioxidant and insulin sensitizers afforded cytoprotection suggest potential therapeutic approaches. PMID- 19807654 TI - The immunosuppressive agent FK506 prevents subperineurial degeneration and demyelination on ultrastructural and functional analysis. AB - Several kinds of injury models, such as crush, transection and graft repair have been well studied in terms of neuroprotective effect of FK506. However, definitive experimental studies are lacking on focal degeneration or ischemia. In the present study, our goal was to investigate the effect of FK506 on functional recovery of the sciatic nerve after focal ischemia, produced by stripping of the epineurial vessels. A total number of 48 Wistar rats were used for this purpose and divided into four groups (control, sham-operated, FK506-treated, and Vehicle treated). Sciatic nerves were approached by femoral and gluteal muscle splitting. Then, epineurial vessels around the sciatic nerve were stripped in the FK506 treated and Vehicle-treated groups. After operation, 5mg/kg/day FK506 administration was initiated by subcutaneous injection until animal sacrifice. The same volume of saline was administrated to the vehicle-treated group. The functional and sensory recoveries were tested by walking pattern analysis and pinch test in every postoperative week. The animals were sacrificed in the end of the fourth postoperative week and sciatic nerve samples were harvested and processed for electron microscopic evaluation. Our data revealed that FK506 administration showed beneficial effect on subperineurial degeneration/demyelinization from functional, sensorial, and ultrastructural points of view. The sciatic nerve samples in the FK506-treated group had several remyelinated fibers compared to the vehicle-treated group. Our literature searches revealed that FK506 administration has not, to our knowledge, been studied in focal ischemic degeneration produced by stripping of the epineurial vessels. PMID- 19807655 TI - Role of endogenous granulocyte-macrophage colony stimulating factor following stroke and relationship to neurological outcome. AB - Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) is a proinflammatory cytokine with neuroprotective and angiogenic properties demonstrated in animal models of cerebral ischemia but their role in human ischemic stroke is still unknown. Thus, our aim is to determine human GM-CSF plasma level in control subjects and stroke patients and its relationship to clinical outcome. Forty three patients with middle cerebral artery occlusion who received thrombolytic therapy within the first three hours of stroke onset and nineteen healthy controls were included. Blood samples were drawn before tissue plasminogen activator (t-PA) treatment. In a group of thirteen strokes blood samples were also obtained one hour after t-PA treatment, at 24 hours of symptoms onset, at discharge and at three months. GM-CSF levels were determined by enzyme-linked immunosorbent assay (ELISA). Stroke severity and neurological outcome was assessed by National Institutes of Health Stroke Scale (NIHSS) and functional outcome were scored by modified Rankin Scale (mRS) at 3 months. Baseline GM-CSF level was significantly higher in stroke patients than in healthy controls (17.8 pg/ml vs 12.8 pg/ml); p<0.0001 and was positively correlated with NIHSS score at 12 hours (R=0.3, p=0.03). No association was detected with functional status at three months measured by mRS. Temporary profile of GM-CSF level in stroke patients gradually decreases from admission to three months. Higher plasma endogenous GM-CSF level is found in stroke patients compared to controls. However, no relation was found with a better outcome. Further research is necessary for elucidating the role of GM-CSF in ischemic stroke. PMID- 19807656 TI - Spatial correlations between the vacuolation, prion protein (PrPsc) deposits and the cerebral blood vessels in sporadic Creutzfeldt-Jakob disease. AB - In the variant form of Creutzfeldt-Jakob disease (vCJD), 'florid' deposits of the protease resistant form of prion protein (PrP(sc)) were aggregated around the cerebral blood vessels suggesting the possibility that prions may spread into the brain via the cerebral microcirculation. The objective of the present study was to determine whether the pathology was spatially related to blood vessels in cases of sporadic CJD (sCJD), a disease without an iatrogenic etiology, and therefore, less likely to be caused by hematogenous spread. Hence, the spatial correlations between the vacuolation ('spongiform change'), PrP(sc) deposits, and the blood vessels were studied in immunolabelled sections of the cerebral cortex and cerebellum in eleven cases of the common M/M1 subtype of sCJD. Both the vacuolation and the PrP(sc) deposits were spatially correlated with the blood vessels; the PrP(sc) deposits being more focally distributed around the vessels than the vacuoles. The frequency of positive spatial correlations was similar in the different gyri of the cerebral cortex, in the upper and lower cortical laminae, and in the molecular layer of the cerebellum. It is hypothesized that the spatial correlation is attributable to factors associated with the blood vessels which promote the aggregation of PrP(sc) to form deposits rather than representing the hematogenous spread of the disease. The aggregated form of PrP(sc) then enhances cell death and may encourages the development of vacuolation in the vicinity of the blood vessels. PMID- 19807658 TI - Oxidative stress-induced necrotic cell death via mitochondira-dependent burst of reactive oxygen species. AB - Oxidative stress is deeply involved in various brain diseases, including neurodegenerative diseases, stroke, and ischemia/reperfusion injury. Mitochondria are thought to be the target and source of oxidative stress. We investigated the role of mitochondria in oxidative stress-induced necrotic neuronal cell death in a neuroblastoma cell line and a mouse model of middle cerebral artery occlusion. The exogenous administration of hydrogen peroxide was used to study the role of oxidative stress on neuronal cell survival and mitochondrial function in vitro. Hydrogen peroxide induced non-apoptotic neuronal cell death in a c-Jun N-terminal kinase- and poly(ADP-ribosyl) polymerase-dependent manner. Unexpectedly, hydrogen peroxide treatment induced transient hyperpolarization of the mitochondrial membrane potential and a subsequent delayed burst of endogenous reactive oxygen species (ROS). The inhibition of mitochondrial hyperpolarization by diphenylene iodonium or rotenone, potent inhibitors of mitochondrial respiratory chain complex I, resulted in reduced ROS production and subsequent neuronal cell death in vitro and in vivo. The inhibition of mitochondrial hyperpolarization can protect neuronal cells from oxidative stress-induced necrotic cell death, suggesting a novel method of therapeutic intervention in oxidative stress-induced neurological disease. PMID- 19807659 TI - Repair of bone defect using bone marrow cells and demineralized bone matrix supplemented with polymeric materials. AB - We present a novel, reverse thermo-responsive (RTR) polymeric osteogenic composite comprising demineralized bone matrix (DBM) and unmanipulated bone marrow cells (BMC) for repair of bone defects. The polymers investigated were low viscosity aqueous solutions at ambient temperature, which gel once they heat up and reach body temperature. Our goal to supplement DBM-BMC composite with RTR polymers displaying superior rheological properties, was to improve graft integrity and stability, during tissue regeneration. The osteogenic composite when implanted under kidney capsule of mice, proved to be biocompatible and biodegradable, with no residual polymer detected in the newly formed osteohematopoietic site. Implantation of the osteogenic composite into a large area of missing area of parietal bone of the skull of rats, resulted in an extensive remodeling of DBM particles, fully reconstituted hematopoietic microenvironment and well integrated normal flat bone within thirty days. The quality and shape of the newly created bone were comparable to the original bone and neither local or systemic inflammatory reactions nor fibrosis at the junction of the new and old calvarium could be documented. Furthermore, combined laser capture microdissection (LCM) technique and PCR analysis of male BMC in female rats confirmed the presence of male derived cells captured from the repaired/ regenerated flat bone defect. The use of active self sufficient osteogenic DBM BMC composite supported by a viscous polymeric scaffold for purposive local hard tissue formation, may have a significant potential in enhancement of bone regeneration and repair following trauma, degenerative or inflamatory lesion, iatrogenic interventions and cosmetic indications. PMID- 19807657 TI - FoxO3a governs early microglial proliferation and employs mitochondrial depolarization with caspase 3, 8, and 9 cleavage during oxidant induced apoptosis. AB - Microglia of the central nervous system have a dual role in the ability to influence the survival of neighboring cells. During inflammatory cell activation, microglia can lead to the disposal of toxic cellular products and permit tissue regeneration, but microglia also may lead to cellular destruction with phagocytic removal. For these reasons, it is essential to elucidate not only the underlying pathways that control microglial activation and proliferation, but also the factors that determine microglial survival. In this regard, we investigated in the EOC 2 microglial cell line with an oxygen-glucose deprivation (OGD) injury model of oxidative stress the role of the "O" class forkhead transcription factor FoxO3a that in some scenarios is closely linked to immune system function. We demonstrate that FoxO3a is a necessary element in the control of early and late apoptotic injury programs that involve membrane phosphatidylserine externalization and nuclear DNA degradation, since transient knockdown of FoxO3a in microglia preserves cellular survival 24 hours following OGD exposure. However, prior to the onset of apoptotic injury, FoxO3a facilitates the activation and proliferation of microglia as early as 3 hours following OGD exposure that occurs in conjunction with the trafficking of the unphosphorylated and active post-translational form of FoxO3a from the cytoplasm to the cell nucleus. FoxO3a also can modulate apoptotic mitochondrial signal transduction pathways in microglia, since transient knockdown of FoxO3a prevents mitochondrial membrane depolarization as well as the release of cytochrome c during OGD. Control of this apoptotic cascade also extends to progressive caspase activation as early as 1 hour following OGD exposure. The presence of FoxO3a is necessary for the expression of cleaved (active) caspase 3, 8, and 9, since loss of FoxO3a abrogates the induction of caspase activity. Interestingly, elimination of FoxO3a reduced caspase 9 activity to a lesser extent than that noted with caspase 3 and 8 activities, suggesting that FoxO3a in relation to caspase 9 may be more reliant upon other signal transduction pathways potentially independent from caspase 3 and 8. PMID- 19807661 TI - Applications of human umbilical cord blood cells in central nervous system regeneration. AB - In recent decades, there has been considerable amount of information about embryonic stem cells (ES). The dilemma facing scientists interested in the development and use of human stem cells in replacement therapies is the source of these cells, i.e. the human embryo. There are many ethical and moral problems related to the use of these cells. Hematopoietic stem cells from umbilical cord blood have been proposed as an alternative source of embryonic stem cells. After exposure to different agents, these cells are able to express antigens of diverse cellular lineages, including the neural type. The In vitro manipulation of human umbilical cord blood (hUCB) cells has shown their stem capacity and plasticity. These cells are easily accessible, In vitro amplifiable, well tolerated by the host, and with more primitive molecular characteristics that give them great flexibility. Overall, these properties open a promising future for the use of hUCB in regenerative therapies for the Central Nervous System (CNS). This review will focus on the available literature concerning umbilical cord blood cells as a therapeutic tool for the treatment of neurodegenerative diseases. PMID- 19807663 TI - A case study from the chemistry core of the Pittsburgh Molecular Library Screening Center: the Polo-like kinase polo-box domain (Plk1-PBD). AB - The Polo-like kinase (Plk) family comprises four cell cycle serine/threonine kinases, Plk1-4. Among these, Plk1 has been most thoroughly characterized; it contains a conserved kinase domain and a C-terminal docking site for S/T phosphorylated proteins (polo-box domain, PBD). Polo-like kinases are deregulated in oncogenesis and therefore constitute a therapeutic target for cancer. A high throughput screening campaign was carried out by the Pittsburgh Molecular Library Screening Center (PMLSC), using a fluorescence polarization assay with recombinant Plk1-PBD to monitor the inhibition of binding of an optimal phosphopeptide substrate motif with recombinant Plk1-PBD. Screening of 97,090 small molecule library samples provided by the NIH Small Molecule Repository distributed by DPI Galapagos led to 11 confirmed hits. The Pittsburgh MLSCN Chemistry Core selected one of the structurally most tractable hits, SID 861574, for chemical hit-to-probe development. A broad chemistry program was initiated that developed new strategies for 6-amino- and 6-hydroxy uracil synthesis as well as acylanilides, and generated a total of 70 analogs. Out of 46 analogues tested, none, nor the resynthesized hit, showed affinity to Plk1-PBD in the follow up assays. In contrast, re-assays of the original screening materials displayed activities similar to the original HTS assay. We ultimately concluded that an impurity in the commercial material led to the positive screening artifact. This case study highlights our development of a synthesis of 6-position functionalized uracil analogs, but also illustrates the importance of careful quality and compound stability monitoring of screening collections. PMID- 19807662 TI - Discovery of selective probes and antagonists for G-protein-coupled receptors FPR/FPRL1 and GPR30. AB - Recent technological advances in flow cytometry provide a versatile platform for high throughput screening of compound libraries coupled with high-content biological testing and drug discovery. The G protein-coupled receptors (GPCRs) constitute the largest class of signaling molecules in the human genome with frequent roles in disease pathogenesis, yet many examples of orphan receptors with unknown ligands remain. The complex biology and potential for drug discovery within this class provide strong incentives for chemical biology approaches seeking to develop small molecule probes to facilitate elucidation of mechanistic pathways and enable specific manipulation of the activity of individual receptors. We have initiated small molecule probe development projects targeting two distinct families of GPCRs: the formylpeptide receptors (FPR/FPRL1) and G protein-coupled estrogen receptor (GPR30). In each case the assay for compound screening involved the development of an appropriate small molecule fluorescent probe, and the flow cytometry platform provided inherently biological rich assays that enhanced the process of identification and optimization of novel antagonists. The contributions of cheminformatics analysis tools, virtual screening, and synthetic chemistry in synergy with the biomolecular screening program have yielded valuable new chemical probes with high binding affinity, selectivity for the targeted receptor, and potent antagonist activity. This review describes the discovery of novel small molecule antagonists of FPR and FPRL1, and GPR30, and the associated characterization process involving secondary assays, cell based and in vivo studies to define the selectivity and activity of the resulting chemical probes. PMID- 19807660 TI - Epigenetic remodeling of chromatin architecture: exploring tumor differentiation therapies in mesenchymal stem cells and sarcomas. AB - Sarcomas are the mesenchymal-derived malignant tumors of connective tissues (e.g., fat, bone, and cartilage) presumed to arise from aberrant development or differentiation of mesenchymal stem cells (MSCs). Appropriate control of stem cell maintenance versus differentiation allows for normal connective tissue development. Current theories suggest that loss of this control--through accumulation of genetic lesions in MSCs at various points in the differentiation process--leads to development of sarcomas, including undifferentiated, high grade sarcoma tumors. The initiation of stem cell differentiation is highly associated with alteration of gene expression, which depends on chromatin remodeling. Epigenetic chromatin modifying agents have been shown to induce cancer cell differentiation and are currently being used clinically to treat cancer. This review will focus on the importance of epigenetic chromatin remodeling in the context of mesenchymal stem cells, sarcoma tumorigenesis and differentiation therapy. PMID- 19807664 TI - The pilot phase of the NIH Chemical Genomics Center. AB - The NIH Chemical Genomics Center (NCGC) was the inaugural center of the Molecular Libraries and Screening Center Network (MLSCN). Along with the nine other research centers of the MLSCN, the NCGC was established with a primary goal of bringing industrial technology and experience to empower the scientific community with small molecule compounds for use in their research. We intend this review to serve as 1) an introduction to the NCGC standard operating procedures, 2) an overview of several of the lessons learned during the pilot phase and 3) a review of several of the innovative discoveries reported during the pilot phase of the MLSCN. PMID- 19807665 TI - Small-molecule modulators of the NF-kappaB pathway newly identified by a translocation-based cellular assay. AB - Nuclear factor kappa B (NF-kappaB) is an important transcription factor. Aberrant regulation of the NF-kappaB pathway is frequently observed in a number of major ailments such as cancer and inflammatory diseases. Hence NF-kappaB modulators have been intensely pursued for their potential therapeutic applications. Numerous reviews have described recent progress in the development of these agents. More recently, a variety of structurally and functionally novel small molecules, identified through high-throughput screens conducted within the Molecular Libraries Screening Center Network (MLSCN) of the NIH Roadmap for Medical Research, have been added to the current list of NF-kappaB regulators. This review will discuss the inhibitors and activators newly discovered by Columbia's Molecular Libraries Screening Center (MLSC) using a well-designed and stable cellular assay. PMID- 19807666 TI - The identification, characterization and optimization of small molecule probes of cysteine proteases: experiences of the Penn Center for Molecular Discovery with cathepsin B and cathepsin L. AB - During the pilot phase of the NIH Molecular Library Screening Network, the Penn Center for Molecular Discovery focused on a series of projects aimed at high throughput screening and the development of probes of a variety of protease targets. This review provides our medicinal chemistry experience with two such targets--cathepsin B and cathepsin L. We describe our approach for hit validation, characterization and triage that led to a critical understanding of the nature of hits from the cathepsin B project. In addition, we detail our experience at hit identification and optimization that led to the development of a novel thiocarbazate probe of cathepsin L. PMID- 19807667 TI - Discovery and development of a potent and highly selective small molecule muscarinic acetylcholine receptor subtype I (mAChR 1 or M1) antagonist in vitro and in vivo probe. AB - This article describes the discovery and development of the first highly selective, small molecule antagonist of the muscarinic acetylcholine receptor subtype I (mAChR1 or M(1)). An M(1) functional, cell-based calcium-mobilization assay identified three distinct chemical series with initial selectivity for M(1) versus M(4). An iterative parallel synthesis approach was employed to optimize all three series in parallel, which led to the development of novel microwave assisted chemistry and provided important take home lessons for probe development projects. Ultimately, this effort produced VU0255035, a potent (IC(50) = 130 nM) and selective (>75-fold vs. M(2)-M(5) and > 10 microM vs. a panel of 75 GPCRs, ion channels and transporters) small molecule M(1) antagonist. Further profiling demonstrated that VU0255035 was centrally penetrant (Brain(AUC)/Plasma(AUC) of 0.48) and active in vivo, rendering it acceptable as both an in vitro and in vivo MLSCN/ MLPCN probe molecule for studying and dissecting M(1) function. PMID- 19807669 TI - Pleiotropic effects of cathepsin D. AB - Over the past decades, the paradigm that lysosomal enzymes participate only in non-specific protein degradation during cell death has changed. Studies conducted both in cell cultures and in animals defined the role of these enzymes that includes cathepsin D (CD). Knockout mice revealed the role of CD in postnatal tissue homeostasis and remodeling. Mutations that abolish the CD enzymatic activity have been implicated in neural ceroid lipofuscinosis. Recent studies suggested a differential role of CD in regulation of apoptosis. The zymogen of CD, procathepsin D (pCD), is secreted by various cancer cells. Extensive studies showed that it acts as a mitogen on both cancer and stromal cells by stimulating their invasive and metastatic properties. Additional studies suggested that procathepsin D/CD is an independent prognostic factor in various cancers, leading to the investigations of pCD/CD as a potential target for designing anti-cancer therapy. In this review, we described the various forms of CD and their implications in numerous physiological as well as pathological conditions. PMID- 19807670 TI - MHC class I TCR engineered anti-tumor CD4 T cells: implications for cancer immunotherapy. AB - T cell immunity is critical for a protective immune response against cancers. Traditionally, this function has been ascribed to CD8 T lymphocytes with cytotoxic activity, which are restricted by MHC class I molecules. The lack of direct cytolytic effector function on part of CD4 T cells, which are MHC class II restricted, coupled with the MHC class II negative nature of most human cancers have been the main reasons for CD8 centered cancer immunotherapy approaches, so far. However, recent findings showing that CD4 T cells play an essential role towards the generation of a productive CD8 response and that the CD4 T cells can also play a direct role in anti-tumor immunity have resulted in growing enthusiasm towards engaging CD4 T cells in cancer immunotherapy. We here discuss the current approaches used for immune based cancer therapy, role of natural MHC class II-restricted CD4 T cells in tumor immunity, factors limiting the engagement of natural CD4 T cells in cancer immunotherapy protocols alongside CD8 T cells, and recent advances in TCR engineering approach to address these limitations. We will also discuss the significance of the MHC class I directed anti-tumor CD4 T cells in tumor immunity. PMID- 19807671 TI - A cytokine-inducing hemagglutinin from small taros. AB - A 22.4-kDa dimeric hemagglutinin was isolated from tubers of Colocasia esculenta cv. 'Small Taro' by employing a purification protocol that involved ion exchange chromatography on Q-Sepharose, fast protein liquid chromatography (FPLC)-ion exchange chromatography on Mono Q, and FPLC-gel filtration on Superdex 75. The hemagglutinin was isolated from the fraction of the taro extract adsorbed on Q Sepharose and subsequently adsorbed on Mono Q. The major absorbance peak from the Superdex 75 column constituted purified hemagglutinin. Its hemagglutinating activity could not be inhibited by simple sugars, and was stable after exposure for 30 minutes to temperatures up to 40 degrees C and to ambient pH in the range of pH 2 to pH 13. The activity decreased progressively when the ambient temperature was raised from 40 degrees C to 100 degrees C. Negligible activity was detected at 100 degrees C. The activity plummeted, with about 40% and 10% remaining, 4 minutes and 20 minutes after exposure to 100 degrees C, respectively. About half of the activity remained at pH 0 and pH 1 whereas the activity was completely abolished at pH 14. The hemagglutinin exhibited slight anti-tumor activity toward hepatoma HepG2 cells, and weak mitogenic activity toward murine splenocytes. It induced expression of the cytokines interleukin-1 beta, interleukin-2, interferon-gamma and tumor necrosis factor-alpha. However, it was devoid of anti-fungal activity toward a number of fungal species. PMID- 19807668 TI - Cancer and virus leads by HTS, chemical design and SEA data mining. AB - A variety of medicinal chemistry approaches can be used for the identification of hits, generation of leads and to accelerate the development of drug candidates. The Emory Chemical and Biology Discovery Center (ECBDC) has been an active participant in the NIH's high-throughput screening (HTS) endeavor to identify potent small molecule probes for poorly studied proteins. Several of Emory's projects relate to cancer or virus infection. We have chosen three successful examples including discovery of potent measles virus RNA-dependent RNA polymerase inhibitors, development of Heat Shock Protein 90 (Hsp90) blockers and identification of angiogenesis inhibitors using transgenic Zebrafish as a HTS model. In parallel with HTS, a unique component of the Emory virtual screening (VS) effort, namely, substructure enrichment analysis (SEA) program has been utilized in several cases. PMID- 19807672 TI - Expression, purification, crystallization and preliminary X-ray crystallographic analysis of the resuscitation promoting factor interacting protein RipA from M. tuberculosis. AB - RipA is an important growth factor of M. tuberculosis. Its depletion produces decreasing bacterial growth and an abnormal phenotype. RipA C-terminal fragment (263-472), containing its predicted catalytic domain has been successfully crystallized using vapor-diffusion methods. The structure has been solved by Multiwavelength Anomalous Dispersion and atomic resolution refinement is in progress. PMID- 19807673 TI - Insight into the stereospecificity of short-chain thermus thermophilus alcohol dehydrogenase showing pro-S hydride transfer and prelog enantioselectivity. AB - The stereochemistry of the hydride transfer in reactions catalyzed by NAD(H) dependent alcohol dehydrogenase from Thermus thermophilus HB27 was determined by means of (1)H-NMR spectroscopy. The enzyme transfers the pro-S hydrogen of [4R (2)H]NADH and exhibits Prelog specificity. Enzyme-substrate docking calculations provided structural details about the enantioselectivity of this thermophilic enzyme. These results give additional insights into the diverse active site architectures of the largely versatile short-chain dehydrogenase superfamily enzymes. A feasible protocol for the synthesis of [4R-(2)H]NADH with high yield was also set up by enzymatic oxidation of 2-propanol-d(8) catalyzed by Bacillus stearothermophilus alcohol dehydrogenase. PMID- 19807674 TI - Purification and characterization of a Laccase with inhibitory activity toward HIV-1 reverse transcriptase and tumor cells from an edible mushroom (Pleurotus cornucopiae). AB - A 66-kDa laccase, with an N-terminal sequence different from those of other mushroom laccases, was purified from fresh fruiting bodies of the edible mushroom Pleurotus cornucopiae by using affinity chromatography on Affi-gel blue gel, ion exchange chromatography on Mono Q and gel filtration on Superdex 75. The procedure resulted in a 16-fold purification and a specific enzyme activity of 17.3 U mg(-1). The optimum pH and temperature for the purified laccase were pH 4 and 40 degrees C, respectively. This laccase inhibited proliferation of murine leukemia cell line L1210 and human hepatoma cell line HepG2, and reduced the activity of HIV-1 reverse transcriptase with an IC50 of 22 microM. There was neither mitogenic activity toward mouse splenocytes, nor hemagglutinating/hemolytic activity toward rabbit erythrocytes. This study yielded information about the potentially exploitable activities of P. cornucopiae laccase. PMID- 19807675 TI - Effect of protein concentration and pH on the chitinase activity of Tapes japonica lysozyme. AB - Tapes japonica lysozyme (TJL), which belongs to the invertebrate-type lysozyme family, has a unique dimer formation. The residues, which include catalytic residues (glutamate 18 and aspartate 30), at the dimer interface form electrostatic interactions. Our previous study suggested that increasing the NaCl concentration switched TJL from a dimer to monomer structure, which increased TJL activity. Therefore, conversion from the dimeric to the monomeric structure is crucial for the TJL activity. In the present study, to further understand the effect of NaCl on TJL dimer formation, we examined the protein concentration and pH dependence of TJL activity in the presence or absence of 500 mM NaCl. TJL activity was suppressed at the high protein concentration. And the optimum pH of TJL activity was decreased in the absence of NaCl. These dependencies confirm the presence of electrostatic interactions between molecules of TJL in the dimeric form in an aqueous solution. PMID- 19807676 TI - Anti-biofilm strategies and the need for innovations in wound care. AB - With an aging and obese population, chronic wounds such as diabetic ulcers, pressure ulcers, and venous leg ulcers are of an increasingly relevant medical concern in the developed world. Identification of bacterial biofilm contamination as a major contributor to non-healing wounds demands biofilm-targeted strategies to treat chronic wounds. While the current standard of care has proven marginally effective, there are components of standard care that should remain part of the wound treatment regime including systemic and topical antibiotics, antiseptics, and physical debridement of biofilm and devitalized tissue. Emerging anti-biofilm strategies include novel, non-invasive means of physical debridement, chemical agent strategies, and biological agent strategies. While aging and obesity will continue to be major burdens to wound care, the emergence of wounds associated with war require investigation and biotechnology development to address biofilm strategies that manage multi-drug resistant bacteria contaminating the chronic wound. The article presents some of the recent patents related to anti-biofilm strategy in wound care. PMID- 19807677 TI - Recent patents in antiviral siRNAs. AB - The advent of gene silencing siRNA technology has created opportunities to develop therapeutics based on targeting the genomics of the disease state. Amongst the first applications of siRNA technology, antiviral applications have been quickly and extensively exploited allowing emergence of a range of antiviral therapeutic strategies. Patent activity has encompassed a range of the components required to utilize this technology ranging from the identification of susceptible genomic targets through to the development of vector systems to express the siRNA endogenously or the synthesis of stable RNA oligonucleotides for in vivo therapeutics. Indeed the primary focus of research effort in this area has been to overcome the challenge common to all of gene therapeutics - delivery of the oligonucleotide - to the diseased tissues and organs, sites of infection and/or sites of drug action. Here we survey the development of siRNA therapeutics both in terms of the range of virus species targeted and the strategic approaches employed. Our study illustrates features commonly observed in the field of nucleic acid drug development. While in vitro studies provide a broad range of molecules and molecular targets for potential therapeutics, the field is however severely limited in terms of safe, effective means to deliver the potential siRNA therapeutics in vivo, to the intracellular site of action. PMID- 19807678 TI - PRO 140--a novel CCR5 co-receptor inhibitor. AB - Despite an increase in the variety of anti-retroviral agents in the market, there remains a need for novel agents to treat HIV 1 infected individuals, in order to overcome existing problems with adherence, toxicities, drug interactions and viral resistance. In this article, we will describe Pro 140, one of the recently developed class of anti-retroviral agent, the CCR5 co-receptor inhibitor. We will also describe several preclinical and clinical studies that have evaluated the efficacy, tolerability and toxicity profiles of Pro-140. We will also look at how its mechanism of action and mode of delivery may change the way patients take highly active anti-retroviral therapy. There are some promising patents discussed in this short review for the use of PRO 140 as CCR5 co-receptor Inhibitor. PMID- 19807679 TI - Recent advances in developing ophthalmic formulations: a patent review. AB - In an effort to improve the drug solubility, stability and/or ocular bioavailability of ophthalmic formulations,various approaches have been explored in the recent past. Additionally, different formulations have been investigated in order to seek those preservative systems that are more tolerable to the ocular tissue. Over the past ten years, inventions in ophthalmic formulations directed toward front-of-eye instillations have concentrated in the areas of new excipients' applications, novel and combined use of conventional excipients, and developments of novel dosage forms. Among these areas, applications of polymeric excipients, cyclodextrins and stabilized chloride dioxide (SCD) have been the most actively studied fields. In addition, oil-in-water (O/W) emulsions have been becoming more popular as an ophthalmic dosage form due to the potentials in increasing drug solubility, stabilizing active pharmaceutical ingredients (APIs), improving ocular tolerance, and providing palliative effects. Some of these innovations from the past decade have the capability of leading to new commercial products. This patent review has a useful knowledge in the advancement for treating various ophthalmic diseases. PMID- 19807680 TI - E-tongue: a tool for taste evaluation. AB - Taste has an important role in the development of oral pharmaceuticals. With respect to patient acceptability and compliance, taste is one of the prime factors determining the market penetration and commercial success of oral formulations, especially in pediatric medicine. Taste assessment is one important quality-control parameter for evaluating taste-masked formulations. Hence, pharmaceutical industries invest time, money and resources into developing palatable and pleasant-tasting products. The primary method for the taste measurement of a drug substance or a formulation is by human sensory evaluation, in which tasting a sample is relayed to inspectors. However, this method is impractical for early stage drug development because the test in humans is expensive and the taste of a drug candidate may not be important to the final product. Therefore, taste-sensing analytical devices, which can detect tastes, have been replacing the taste panelists. In the present review we are presenting different aspect of electronic tongue. The review article also discussed some useful patents and instrument with respect to E-tongue. PMID- 19807681 TI - Microemulsions as carriers for therapeutic molecules. AB - The thrust for finding newer drug delivery systems for exiting therapeutic molecules has opened a wide window for colloidal systems. Due to the presence of different domains of variable polarity in the microemulsion systems, they show a huge potential to be used as drug delivery vehicles for a variety of drugs. The use of microemulsion as drug delivery vehicles through a number of routes has engaged a large number of research groups in this area. Microemulsion media finds several applications ranging from drug delivery to drug nanoparticle templating due to its ability to enhance solubility, stability and bioavailability. This review on patent articles recounts the patent literature dealing with different kind of microemulsion carriers used via different routes, solubility and permeability enhancement and its use as a template for nanoparticle synthesis. PMID- 19807682 TI - Microporation techniques for enhanced delivery of therapeutic agents. AB - Perhaps the greatest barrier to development of the field of transmembrane drug delivery is that only a limited number of drugs are amenable to administration by this route. The highly lipophilic nature and barrier function of the uppermost layer of the skin, the stratum corneum, for example, restricts the permeation of hydrophilic, high molecular weight and charged compounds into the systemic circulation. Other membranes in the human body can also present significant barriers to drug permeation. In order to successfully deliver hydrophilic drugs, and macromolecular agents of interest, including peptides, DNA and small interfering RNA, many research groups and pharmaceutical companies Worldwide are focusing on the use of microporation methods and devices. Whilst there are a variety of microporation techniques, including the use of laser, thermal ablation, electroporation, radiofrequency, ultrasound, high pressure jets, and microneedle technology, they share the common goal of enhancing the permeability of a biological membrane through the creation of transient aqueous transport pathways of micron dimensions across that membrane. Once created, these micropores are orders of magnitude larger than molecular dimensions and, therefore, should readily permit the transport of hydrophilic macromolecules. Additionally, microporation devices also enable minimally-invasive sampling and monitoring of biological fluids. This review deals with the innovations relating to microporation-based methods and devices for drug delivery and minimally invasive monitoring, as disclosed in recent patent literature. PMID- 19807683 TI - Drug delivery systems for local anesthetics. AB - Although technological innovations in the area of drug delivery claim for varied benefits, increasing the drug therapeutic index for human clinical application is the main goal pursued. Drug delivery systems for local anesthetics (LA) have attracted researchers due to many biomedical advantages associated to their application. Formulation approaches to systemically deliver LA include the encapsulation in liposomes, complexation in cyclodextrins, association with biopolymers and others carrier systems. Topical delivery systems for LA are characteristically composed by a diversity of adjuvants (viscosity inducing agents, preservatives, permeation enhancers, emollients,) and presentations such as semisolid (gel, creams, ointments), liquid (o/w and w/o emulsions, dispersions) and solid (patches) pharmaceutical forms. The proposed formulations aims to reduce the LA concentration used, increase its permeability and absorption, keep the LA at the target site for longer periods prolonging the anesthetic or analgesic effect and, finally, to decrease the clearance, local and systemic toxicity. This review deals with the innovations pertaining to formulations and techniques for drug-delivery of topical and injectable local anesthetics, as described in recent patents. PMID- 19807684 TI - Barminomycin, a model for the development of new anthracyclines. AB - Barminomycin is a member of the anthracycline class of anticancer agents and was originally discovered as a pink/red complex with DNA and RNA and named SN-07. The chromophore was subsequently separated from the nucleic acids by nuclease digestion and contained the four-membered anthraquinone ring system characteristic of anthracyclines, but with an unusual eight membered ring that contained a carbinolamine which readily interconverted to an imine. The imine form is analogous to the formaldehyde-activated form of other anthracyclines such as doxorubicin. The imine form confers exceptional activity to barminomycin which is 1,000-fold more cytotoxic than doxorubicin. Barminomycin rapidly forms adducts with DNA, reacting with the exocyclic amino group of guanine residues and with high selectivity for 5'-GC-3' sequences. The coupling to DNA appears to be identical to the N-C-N aminal linkage formed between doxorubicin and DNA where the carbon derives from formaldehyde for doxorubicin-DNA adducts, whereas this "activated carbon" is an inherent component of the imine group in the eight membered ring of barminomycin. Although the linkage of both drugs to DNA appears to be identical, barminomycin-DNA complexes are essentially irreversible compared to the labile doxorubicin-DNA adducts which have an in vitro (purified DNA) half life of 25 h at 37 degrees C. A 3D model of the barminomycin-DNA complex has been defined from 307 NOE distance constraints. The enhanced stability of barminomycin DNA adducts appears to be due primarily to protection of the aminal linkage from hydrolysis and this has provided insight into the design of new anthracycline derivatives with enhanced stability and activity. Strategies for harnessing the extreme reactivity and activity of barminomycin are also presented. PMID- 19807685 TI - Mitochondrially targeted antioxidants for the treatment of cardiovascular diseases. AB - Oxidative stress resulting from imbalance between reactive oxygen species (ROS) generation and antioxidant mechanisms is important in the pathogenesis of cardiovascular diseases such as atherosclerosis, ischemic heart disease, heart failure, stroke, hypertension and diabetes. Paradoxically, antioxidant therapy such as vitamin E has not been shown on large randomized clinical trials to favorably affect clinical outcomes. Since mitochondria are involved not only with bioenergetics but also with oxidative damage through ROS generation and cell signaling leading to apoptosis, antioxidants targeted at the mitochondria are appealing novel agents to attenuate oxidative stress. In particular, antioxidants conjugated with triphenylphosphonium cation such as mitoquinone, mitovitamin E and mitophenyltertbutyline achieve concentrations in the mitochondrial matrix several-fold greater than those achieved in the cytosol because of the high negative membrane potential of the inner mitochondrial membrane. We review preliminary experiments and also some patents on cell and animal models of cardiovascular diseases where mitochondrially targeted antioxidants have been used and were shown to reduce ROS production and the effects of oxidative stress due to ROS, apoptosis and improve cardiac function. Although ongoing human clinical studies involve only non-cardiovascular applications at this time, preclinical studies show promise for eventual human trials for cardiovascular diseases. PMID- 19807686 TI - Major distractions; innervational infarction disruption of binocular vision; spectacle correction of hyperopia in esotropia; non ocular dominance; semantics euphemisms! economize = compete = true "reform" versus healthcare warfare? to our end? by our self annointed royalty (a veritable tribe of kings). PMID- 19807687 TI - Cycloplegic refractions in children who never wore and who always wore prescribed spectacles for refractive accommodative esotropia: exploring the natural history of this form of strabismus and the effect of treatment on their hyperopia. AB - PURPOSE: To compare cycloplegic refractions in accommodative esotropes who never and who always wore prescribed spectacles. METHODS: An institutional retrospective medical record review was performed for children with fully or partially refractive accommodative esotropia without neurological or other ocular disease. Only children with a least 3 years (y) follow-up, at least 2 separate cycloplegic refractions at least 3y apart, and clear documentation of full compliance and conplete non-complicance with prescribed spectacles were studied. RESULTS: For the right eye, average youngest (less than 2y) and oldest (8-10Y) spherical equivalents were significantly lower in non-compliant children (3.3D [n=25] and 2.49D [n=13]) than in compliant children (5.5D [n=8] and 4.69D [n=25]). The differences in mean hyperopia between less than 2y and 8-10y for the non-compliant (-0.81D) and compliant (-0.84D) children were similar as were levels of mean cylinder. CONCLUSIONS: The differences in average spherical equivalent and average degree of astigmatism between less than 2y and 8-10 y (slight decrease and slight increase, respectively were similar for children who never and children who always wore prescribed glasses for refractive accommodatiave esotropia. Although full-time glasses wear did not appear to affect refractive shifts, limitations of this retrospective study include a lack of statistical power to detect differences less than 2D. PMID- 19807688 TI - Effects of innervational strabismus surgery on a patient with hypotropic strabismus and pseudoblepharoptosis. AB - PURPOSE: We seek to analyze the results of the innervational surgery in a case of hypotropia with peudoptosis with vidiooculography (3D-VOG). CASE REPORT: We present the clinical case of a 35 year old male suffereing from acquired ptosis of the left eyelid, present since the age of 10 and who was interested in cosmetic surgery. The preoperative 3D-VOG shows hypotropia of the left eye (22.5 degrees), minimum exotropia and intorsion together with limitation in the elevation of the left eye with positive passive duction test and pseudoptosis. RESULTS: Surgery was carried out in three stages: 1. Recession of the superior rectus of the right eye (RE) resulting in an improvement in the elevation and the pseudoptosis; 2. Recession of the inferior rectus of the left eye (LE), with improvement in the elevation of the LE but deterioration of the pseudoptosis; 3. Resection of the inferior rectus of the RE which improves both the elevation as well as the pseudoptosis of the LE. CONCLUSIONS: We consider the usefulness of the innervational techniques in the incomplete third nerve paralysis with important affectation in the elevation, making it necessary to check and in this case eliminate the restrictive effects although this may influence the effect on the pseudoptosis. PMID- 19807689 TI - Partial third nerve palsy involving the superior rectus and levator palpebrae muscles and distruption of central binocular vision fusion from brain stem infarction: a case report. AB - BACKGROUND AND PURPOSE: To add to the literature of a case of isolated third nerve paresis involving the nerve fascicles subserving the superior rectus and the levator palpebrae muslces from brain stem infarction and presenting the characteristics of central disruption of binocular vision fusional amplitudes. CASE REPORT: One patient with an old intracranial aneurysm and with old and recent brain stem infarcts and no other neurological manifestations, demonstrating findings characteristic of isolated paresis of the superior rectus and levator palpebrae muscles is reported. CONCLUSION: This dual involvement of the superior rectus and levator palpebrae muscles supports the anatomical arrangement of the ocular motor nucleus fascicles in the midbrain, clarified by experimental studies on animals and clinical data in humans and emphasizes the juxtaposition of the superior rectus and levator palpebrae fascicles and placing the levator palpebrae mucle fascicle lateral to the medial rectus fascicle in the midbrain. The comitant vertical deviation and the negative Bielschowsky head tilt test support the vertical rectus muscle involvement. The constant diplopia with only 4 prism diopters of hypotropia and with the absence of fusional amplitudes evokes disruption of central binocular fusion. PMID- 19807690 TI - A precise and rapid mapping protocol for correlative light and electron microscopy of small invertebrate organisms. AB - BACKGROUND INFORMATION: CLEM (correlative live cell and electron microscopy) seeks to bridge the data acquired with different imaging strategies, typically between light microscopy and electron microscopy. It has been successfully applied in cell cultures, although its use in multicellular systems is hampered by difficulties in locating the ROI (region of interest). RESULTS: We developed a CLEM technique that enables easy processing of small model animals and is adequate both for morphology and immunoelectron-microscopic specimen preparations. While this method has been initially developed for Caenorhabditis elegans samples, we found that it works equally well for Drosophila samples. It enables handling and observation of single animals of any complex genotype in real time, fixation by high-pressure freezing and flat embedding. Our major improvement has been the development of a precise mapping system that considerably simplifies and speeds up the retrospective location of the ROI within 1 mum distance. This method can be successfully used when correlative microscopy is required, as well as to facilitate the treatment of non-correlative TEM procedures. Our improvements open the possibility to treat statistically significant numbers of animals processed by electron microscopy and considerably simplifies electron-microscopic protocols, making them more accessible to a wider range of researchers. CONCLUSIONS: We believe that this technique will contribute to correlative studies in multicellular models and will facilitate the time demanding procedure of specimen preparation for any kind of TEM. PMID- 19807692 TI - Regulatory factors controlling transcription of Saccharomyces cerevisiae IXR1 by oxygen levels: a model of transcriptional adaptation from aerobiosis to hypoxia implicating ROX1 and IXR1 cross-regulation. AB - Ixr1p from Saccharomyces cerevisiae has been previously studied because it binds to DNA containing intrastrand cross-links formed by the anticancer drug cisplatin. Ixr1p is also a transcriptional regulator of anaerobic/hypoxic genes, such as SRP1/TIR1, which encodes a stress-response cell wall manoprotein, and COX5B, which encodes the Vb subunit of the mitochondrial complex cytochrome c oxidase. However, factors controlling IXR1 expression remained unexplored. In the present study we show that IXR1 mRNA levels are controlled by oxygen availability and increase during hypoxia. In aerobiosis, low levels of IXR1 expression are maintained by Rox1p repression through the general co-repressor complex Tup1 Ssn6. Ixr1p itself is necessary for full IXR1 expression under hypoxic conditions. Deletion analyses have identified the region in the IXR1 promoter responsible for this positive auto-control (nucleotides -557 to -376). EMSA (electrophoretic mobility-shift assay) and ChIP (chromatin immunoprecipitation) assays show that Ixr1p binds to the IXR1 promoter both in vitro and in vivo. Ixr1p is also required for hypoxic repression of ROX1 and binds to its promoter. UPC2 deletion has opposite effects on IXR1 and ROX1 transcription during hypoxia. Ixr1p is also necessary for resistance to oxidative stress generated by H2O2. IXR1 expression is moderately activated by H2O2 and this induction is Yap1p dependent. A model of IXR1 regulation as a relay for sensing different signals related to change in oxygen availability is proposed. In this model, transcriptional adaptation from aerobiosis to hypoxia depends on ROX1 and IXR1 cross-regulation. PMID- 19807691 TI - Molecular identification of aspartate N-acetyltransferase and its mutation in hypoacetylaspartia. AB - The brain-specific compound NAA (N-acetylaspartate) occurs almost exclusively in neurons, where its concentration reaches approx. 20 mM. Its abundance is determined in patients by MRS (magnetic resonance spectroscopy) to assess neuronal density and health. The molecular identity of the NAT (N acetyltransferase) that catalyses NAA synthesis has remained unknown, because the enzyme is membrane-bound and difficult to purify. Database searches indicated that among putative NATs (i.e. proteins homologous with known NATs, but with uncharacterized catalytic activity) encoded by the human and mouse genomes two were almost exclusively expressed in brain, NAT8L and NAT14. Transfection studies in HEK-293T [human embryonic kidney-293 cells expressing the large T-antigen of SV40 (simian virus 40)] indicated that NAT8L, but not NAT14, catalysed the synthesis of NAA from L-aspartate and acetyl-CoA. The specificity of NAT8L, its Km for aspartate and its sensitivity to detergents are similar to those described for brain Asp-NAT. Confocal microscopy analysis of CHO (Chinese-hamster ovary) cells and neurons expressing recombinant NAT8L indicates that it is associated with the ER (endoplasmic reticulum), but not with mitochondria. A mutation search in the NAT8L gene of the only patient known to be deficient in NAA disclosed the presence of a homozygous 19 bp deletion, resulting in a change in reading frame and the absence of production of a functional protein. We conclude that NAT8L, a neuron-specific protein, is responsible for NAA synthesis and is mutated in primary NAA deficiency (hypoacetylaspartia). The molecular identification of this enzyme will lead to new perspectives in the clarification of the function of this most abundant amino acid derivative in neurons and for the diagnosis of hypoacetylaspartia in other patients. PMID- 19807693 TI - Coactosin-like protein functions as a stabilizing chaperone for 5-lipoxygenase: role of tryptophan 102. AB - The activity of 5-LO (5-lipoxygenase), which catalyses two initial steps in the biosynthesis of pro-inflammatory LTs (leukotrienes), is strictly regulated. One recently discovered factor, CLP (coactosin-like protein), binds 5-LO and promotes LT formation. In the present paper we report that CLP also stabilizes 5-LO and prevents non-turnover inactivation of the enzyme in vitro. Mutagenesis of tryptophan residues in the 5-LO beta-sandwich showed that 5-LO-Trp102 is essential for binding to CLP, and for CLP to support 5-LO activity. In addition, the stabilizing effect also depended on binding between CLP and 5-LO. After mutations which prevent interaction (5-LO-W102A or CLP-K131A), the protective effect of CLP was absent. A calculated 5-LO-CLP docking model indicates that CLP may bind to additional residues in both domains of 5-LO, thus possibly stabilizing the 5-LO structure. To obtain further support for binding between CLP and 5-LO in a living cell, subcellular localization of CLP and 5-LO in the monocytic cell line Mono Mac 6 was determined. In these cells, 5-LO associates with a nuclear fraction only when differentiated cells are primed with phorbol ester and stimulated with ionophore. The same pattern of redistribution was found for CLP, indicating that the two proteins associate with the nucleus in a co ordinated fashion. The results of the present study support a role for CLP as a chaperoning scaffold factor, influencing both the stability and the activity of 5 LO. PMID- 19807694 TI - Targeted ablation and reorganization of the principal preplate neurons and their neuroblasts identified by golli promoter transgene expression in the neocortex of mice. AB - The present study delineates the cellular responses of dorsal pallium to targeted genetic ablation of the principal preplate neurons of the neocortex. Ganciclovir treatment during prenatal development (E11-E13; where E is embryonic day) of mice selectively killed cells with shared S-phase vulnerability and targeted expression of a GPT [golli promoter transgene, linked to HSV-TK (herpes simplex virus-thymidine kinase), tau-eGFP (tau-enhanced green fluorescent protein) and lacZ (lacZ galactosidase) reporters] localized in preplate neurons. Morphogenetic fates of attacked neurons and neuroblasts, and their successors, were assessed by multiple labelling in time-series comparisons between ablated (HSV-TK+/0) and control (HSV-TK0/0) littermates. During ablation generation, neocortical growth was suppressed, and compensatory reorganization of non-GPT ventricular zone progenitors of dorsal pallium produced replacements for killed GPT neuroblasts. Replacement and surviving GPT neuroblasts then produced replacements for killed GPT neurons. Near-normal restoration of their complement delayed the settlement of GPT neurons into the reconstituted preplate, which curtailed the outgrowth of pioneer corticofugal axons. Based on this evidence, we conclude that specific cell killing in ablated mice can eliminate a major fraction of GPT neurons, with insignificant bystander killing. Also, replacement GPT neurons in ablated mice originate exclusively by proliferation from intermediate progenitor GPT neuroblasts, whose complement is maintained by non-GPT progenitors for inductive regulation of the total complement of GPT neurons. Finally, GPT neurons in both normal and ablated mice meet all morphogenetic criteria, including the 'outside in' vertical gradient of settlement, presently used to identify principal preplate neurons. In ablated mice, delayed organization of these neurons desynchronizes and isolates developing neocortex from the rest of the brain, and permanently impairs its connectivity. PMID- 19807695 TI - Are stem cells a cure for diabetes? AB - With the already heightened demand placed on organ donation, stem cell therapy has become a tantalizing idea to provide glucose-responsive insulin-producing cells to Type 1 diabetic patients as an alternative to islet transplantation. Multiple groups have developed varied approaches to create a population of cells with the appropriate characteristics. Both adult and embryonic stem cells have received an enormous amount of attention as possible sources of insulin-producing cells. Although adult stem cells lack the pluripotent nature of their embryonic counterparts, they appear to avoid the ethical debate that has centred around the latter. This may limit the eventual application of embryonic stem cells, which have already shown promise in early mouse models. One must also consider the potential of stem cells to form teratomas, a complication which would prove devastating in an immunologically compromised transplant recipient. The present review looks at the progress to date in both the adult and embryonic stem cells fields as potential treatments for diabetes. We also consider some of the limitations of stem cell therapy and the potential complications that may develop with their use. PMID- 19807696 TI - Complement activation and disease: protective effects of hyperbilirubinaemia. AB - Complement, an important effector mechanism of the immune system, is an enzymatic cascade of approx. 30 serum proteins leading to the amplification of a specific humoral response. It can be activated through the classical or alternative pathways, or through the mannose-binding lectin pathway. The activation of the classical pathway is initiated by the binding of the C1 component to antigen bound antibodies, known as immunocomplexes. C1 is a complex of one molecule of C1q, two molecules of C1r and two molecules of C1s. C1q contains three copies of a Y-shaped fundamental unit with globular heads included in its structure, which play a major role in the interaction with the Fc portion of immunoglobulins. Deficient or exacerbated activation of the complement system leads to diseases of variable severity, and pharmacological inhibition of the complement system is considered as a therapeutic strategy to ameliorate the inflammatory effects of exacerbated complement activation. Bilirubin is a product of haem degradation by the concerted action of haem oxygenase, which converts haem into biliverdin, and biliverdin reductase, which reduces biliverdin to UCB (unconjugated bilirubin). UCB exerts both cytoprotective and cytotoxic effects in a variety of tissues and cells, acting either as an antioxidant at low concentrations or as an oxidant at high concentrations. In the present review, we describe in detail the anti complement properties of bilirubin, occurring at levels above the UCB concentrations found in normal human serum, as a beneficial effect of potential clinical relevance. We provide evidence that UCB interferes with the interaction between C1q and immunoglobulins, thus inhibiting the initial step in the activation of complement through the classical pathway. A molecular model is proposed for the interaction between UCB and C1q. PMID- 19807697 TI - Think the impossible: beta-blockers for treating asthma. AB - Asthma was originally thought to be associated with an intrinsic defect in beta2ADR (beta2-adrenoceptor) function, tipping the balance towards parasympathetic bronchoconstriction. Hence beta-blocking drugs (such as beta2ADR antagonists and inverse agonists) may cause acute bronchoconstriction which, in turn, may be attenuated by anti-cholinergic agents. Although beta2-agonists are highly effective for the acute relief of bronchoconstriction, their chronic use is accompanied by an adaptive reduction in beta2ADR numbers and associated desensitization of response, resulting in increased exacerbations and rare cases of death. The hypothesis examined in the present article is that, while single dosing with a beta-blocker may cause acute bronchoconstriction, chronic dosing may afford putative beneficial effects including attenuated airway hyperresponsiveness. PMID- 19807698 TI - The hepatitis C virus and its hepatic environment: a toxic but finely tuned partnership. AB - Twenty years after its discovery, HCV (hepatitis C virus) still infects 170 million people worldwide and cannot be properly treated due to the lack of efficient medication. Its life cycle must be better understood to develop targeted pharmacological arsenals. HCV is an enveloped virus bearing two surface glycoproteins, E1 and E2. It only infects humans through blood transmission, and hepatocytes are its only target cells. Hepatic trabeculae are formed by hepatocyte rows surrounded by sinusoid capillaries, irrigating hepatic cells. Hepatocytes are polarized and have basolateral and apical poles, separated by tight junctions in contact with blood and bile respectively. In blood, HCV remains in contact with lipoproteins. It then navigates through hepatic microenvironment and extracellular matrix, composed of glycosaminoglycans and proteins. HCV then encounters the hepatocyte basolateral membrane, where it interacts with its entry factors: the low-density lipoprotein receptor, CD81 tetraspanin, and the high-density lipoprotein (scavenger) receptor SR-BI (scavenger receptor BI). How these molecules interact with HCV remains unclear; however, a tentative sequence of events has been proposed. Two essential factors of HCV entry are the tight junction proteins claudin-1 and occludin. Cell polarity therefore seems to be a key for HCV entry. This raises several exciting questions on the HCV internalization pathway. Clathrin-dependent endocytosis is probably the route of HCV transport to intracellular compartments, and the ultimate step of its entry is fusion, which probably takes place within endosomes. The mechanisms of HCV membrane fusion are still unclear, notably the nature of the fusion proteins is unknown and the contribution of HCV-associated lipoproteins to this event is currently under investigation. PMID- 19807700 TI - Early iron supplementation for very low birth weight preterm newborns: statistical vs. clinical significance!! PMID- 19807701 TI - Vesicular eruption located on sunburned areas in an 8-year-old girl. PMID- 19807702 TI - Social impairment due to extreme photophobia. PMID- 19807703 TI - Lung liquid transport components in human perinatal respiratory distress. PMID- 19807704 TI - SIDS: past, present and future. AB - Despite the large reduction in SIDS mortality, which occurred in the early 1990s following the 'Back to Sleep' campaigns, SIDS remains the leading cause of death in the postneonatal age group. This paper describes the position in the 1980s, the contribution of the New Zealand Cot Death Study, what should be recommended and the current research priorities. CONCLUSION: SIDS is preventable. Application of what we currently know could eliminate SIDS. The challenge is to find ways of implementing our knowledge. PMID- 19807705 TI - Overuse injuries in the young athlete. PMID- 19807707 TI - Age of sitting unsupported and independent walking in very low birth weight preterm infants with normal motor development at 2 years. AB - AIMS: The aims of this study is to (i) determine the age of sitting unsupported and independent walking in preterm infants with birth weight under 1500 g (very low birth weight, VLBW); (ii) estimate differences between VLBW children and a reference population and (iii) estimate the association between clinical characteristics and late age at sitting and walking. METHODS: A longitudinal study was conducted of a cohort of 876 children with VLBW. The World Health Organization (WHO) motor development study population was used as a reference. Ages for both skills were established by interview with parents. Means were compared with t-test, ANOVA and Bonferroni adjustment where appropriate. RESULTS: The inclusion criteria were complied with 694 patients; 50% of VLBW sat at 7 m corrected age (CA) and walked at 13 m CA. Both motor skills were acquired later (7.3 +/- 1.5 and 13.6 +/- 2.8 m) compared with the control group (6 +/- 1.1 and 12.1 +/- 1.8 m). Weight or head circumference at birth below the 10th percentile or the presence of bronchopulmonary dysplasia were associated with delayed acquisition of both skills. CONCLUSION: Very low birth weight infants typically sit unsupported and walk later than term infants. Tables describing reference values for milestones acquisition for different categories of infants (gestational age, birth weight and other determinants) may contribute to inform the decision making process on access to available resources. PMID- 19807706 TI - Antioxidant capacity of human milk and its association with vitamins A and E and fatty acid composition. AB - AIM: The antioxidant capacity of human milk reflects the presence and activity of multiple components, which prevent oxidative rancidity. The aim of this study was to use the Oxygen Radical Absorbance Capacity assay to assess human milk antioxidant capacity and find correlations with milk components. METHODS: Milk samples collected from 60 breastfeeding women at 1 month postpartum were assayed for antioxidant capacity, vitamins E and A, and fatty acids. Potential statistical relationships of concentrations of vitamins A and E and polyunsaturated fatty acids on the antioxidant capacity of human milk were determined. RESULTS: Human milk antioxidant capacity was positively attributed to alpha-tocopherol concentration (rho < 0.05). The vitamin A concentration did not significantly contribute to milk antioxidant capacity, but was correlated to milk alpha-tocopherol concentration (r = 0.587; rho < 0.001). There was no evidence of an inverse relationship between polyunsaturated fatty acids concentration and the antioxidant capacity value of milk. CONCLUSION: This study shows that alpha tocopherol is an important contributor to the oxidative stability of human milk. Moreover, there was no evidence obtained to show that women who have high levels of milk polyunsaturated fatty acids are predisposed to lower milk antioxidant capacity. PMID- 19807711 TI - Is the hope for a cellulosic biofuel a lot of rot? PMID- 19807712 TI - Degradation of alkanes by bacteria. AB - Pollution of soil and water environments by crude oil has been, and is still today, an important problem. Crude oil is a complex mixture of thousands of compounds. Among them, alkanes constitute the major fraction. Alkanes are saturated hydrocarbons of different sizes and structures. Although they are chemically very inert, most of them can be efficiently degraded by several microorganisms. This review summarizes current knowledge on how microorganisms degrade alkanes, focusing on the biochemical pathways used and on how the expression of pathway genes is regulated and integrated within cell physiology. PMID- 19807714 TI - Stromgren remembered. PMID- 19807715 TI - Schizophrenia; from structure to function with special focus on the mediodorsal thalamic prefrontal loop. AB - OBJECTIVE: To describe structural and biochemical evidence from postmortem brains that implicates the reciprocal connections between the mediodorsal thalamic nucleus and the prefrontal cortex in cognitive symptoms of schizophrenia. METHOD: The estimation of the regional volumes and cell numbers was obtained using stereological methods. The biochemical analyses of molecular expression in postmortem brain involve quantitative measurement of transcripts and proteins by in-situ (RNA) or Western blot/autoradiography in brains from patients with schizophrenia and comparison subjects. RESULTS: Stereological studies in postmortem brain from patients with schizophrenia have reported divergent and often opposing findings in the total number of neurons and volume of the mediodorsal (MD) thalamic nucleus, and to a lesser degree in its reciprocally associated areas of the prefrontal cortex. Similarly, quantitative molecular postmortem studies have found large inter-subject and between-study variance at both the transcript and protein levels for receptors and their interacting molecules of several neurotransmitter systems in these interconnected anatomical regions. Combined, large variation in stereological and molecular studies indicates a complex and heterogeneous involvement of the MD thalamic-prefrontal loop in schizophrenia. CONCLUSION: Based on a considerable heterogeneity in patients suffering from schizophrenia, large variation in postmortem studies, including stereological and molecular postmortem studies of the MD thalamus and frontal cortex, might be expected and may in fact partly help to explain the variable endophenotypic traits associated with this severe psychiatric illness. PMID- 19807716 TI - From age correction to genome-wide association. AB - OBJECTIVE: Eric Stromgren was one of the pioneers of psychiatric genetics and family studies. There has now been an explosion of interest in this field and research progress, including linkage and association studies, whole genome genotyping, copy number variants and epigenetics is reviewed here. METHOD: An overview of this area of psychiatric research is presented and discussed based on the relevant literature aiming at giving a recent status of the progress. RESULTS: Broadly speaking linkage and association are complementary approaches used to locate genes contributing to the genetic aetiology of psychopathology. Linkage can be detected over comparatively large distances, however power is problematic when searching for quantitative trait loci with small effect sizes. In contrast, association studies can detect small effects but only over very small distances. Therefore, while several genome-wide linkage studies in psychiatric disorders have been performed, the majority of association studies have investigated specific functional candidate genes. CONCLUSION: Due to very recent technological advancements, genome-wide association studies have now become possible and have identified some completely novel susceptibility loci. Other recent advances include the discovery of epigenetic phenomena and copy number variants. PMID- 19807717 TI - 'Salience syndrome' replaces 'schizophrenia' in DSM-V and ICD-11: psychiatry's evidence-based entry into the 21st century? AB - OBJECTIVE: Japan was the first country to abandon the 19th century term of 'mind splitting disease' (schizophrenia). Revisions of DSM and ICD are forthcoming. Should the rest of the world follow Japan's example? METHOD: A comprehensive literature search was carried out in order to review the scientific evidence for the validity, usefulness and acceptability of current concepts of psychotic disorder. RESULTS: The discussion about re-classifying and renaming schizophrenia and other psychotic disorders is clouded by conceptual confusion. First, it is often misunderstood as a misguided attempt to change societal stigma instead of an attempt to change iatrogenic stigma occasioned by the use of misleading and mystifying terminology. Second, the debate is misunderstood as purely semantic, whereas in actual fact it is about the core concepts underlying psychiatric nosology. Third, it has been suggested that the debate is political. However, solid scientific evidence pointing to the absence of nosological validity of diagnostic categories lies at the heart of the argument. Fourth, there is confusion about what constitutes a syndrome (a group of symptom dimensions that cluster in different combinations in different people and for which one or more underlying diseases may or may not be found) and a disease (a nosologically valid entity with specific causes, symptoms, treatment and course). CONCLUSION: Scientific evidence favours a syndromal system of classification combining categorical and dimensional representations of psychosis. The concept of 'salience' has the potential to make the public recognize psychosis as relating to an aspect of human mentation and experience that is universal. It is proposed to introduce, analogous to the functional-descriptive term 'Metabolic syndrome', the diagnosis of 'Salience syndrome' to replace all current diagnostic categories of psychotic disorders. Within Salience syndrome, three subcategories may be identified, based on scientific evidence of relatively valid and specific contrasts, named Salience syndrome with affective expression, Salience syndrome with developmental expression and Salience syndrome not otherwise specified. PMID- 19807718 TI - Psychotherapy of borderline personality disorder. AB - OBJECTIVE: Psychotherapy is considered the primary treatment for borderline personality disorder (BPD). Currently, there are four comprehensive psychosocial treatments for BPD. Two of these treatments are considered psychodynamic in nature: mentalization-based treatment and transference-focused psychotherapy. The other two are considered to be cognitive-behavioral in nature: dialectical behavioral therapy and schema-focused therapy. METHOD: A review of the relevant literature was conducted. RESULTS: Each of these lengthy and complex psychotherapies significantly reduces the severity of borderline psychopathology or at least some aspects of it, particularly physically self-destructive acts. CONCLUSION: Comprehensive, long-term psychotherapy can be a useful form of treatment for those with BPD. However, less intensive and less costly forms of treatment need to be developed. PMID- 19807719 TI - Lithium response across generations. AB - OBJECTIVE: To describe and integrate observations from bipolar patients responsive to lithium stabilization and their children. METHOD: Selected findings are described from the clinical and biological investigations of adults meeting research criteria for bipolar disorder and for responsiveness to lithium stabilization; and from prospective studies of the children of lithium responders and non-responders. RESULTS: Response to prophylactic lithium identifies a valid subtype of bipolar disorder, however the search for biological markers of lithium response, while promising, has so far remained inconclusive. Adult responders to lithium stabilization exhibit definable clinical features which are also observable in their affected children. In prospective studies the children of bipolar parents develop symptoms earlier than reported previously, with marked differences between the offspring of lithium responders and non-responders. The illness evolves in predictable clinical stages, first from non-specific sleep and anxiety disorders to mood symptoms and then, often starting in adolescence, major depressive and later activated episodes. CONCLUSION: Investigating and comparing unequivocal responders and non-responders to long-term lithium treatment and their offspring is a fertile research strategy for addressing a multitude of clinical and research questions. PMID- 19807720 TI - The use of population based registers in psychiatric research. AB - OBJECTIVE: Much of the knowledge we now take for granted regarding major mental disorders such as schizophrenia, suicide and other disorders, would not exist without the use of population based registers. The use of population based registers in psychiatric epidemiology have enabled analyses of associations that otherwise would not have been possible to address. METHOD: The use of registers in psychiatric research is described, exemplified, and discussed. RESULTS: Methodological and validity aspects depend to a large part on the type of register being considered. A classification is proposed of different types of registers, each one implying specific methodological issues. These can be addressed according to the dimensions coverage, attrition, representativity and validity. Specific methodological consideration has still to be taken in relation to each specific research question. Thus, special validity studies usually need to be performed when embarking on studies using population based registers. CONCLUSION: With increasing burden of disease due to mental disorders worldwide, knowledge of the epidemiology of these disorders are of increasing interest. The Nordic countries have a strong history in this field of research, of great interest to the rest of the world. Universities and research funding agencies should recognize this valuable source of research capacity, and support fruitful continuation of a strong tradition. PMID- 19807721 TI - The heterogeneity of causes and courses of attention-deficit/hyperactivity disorder. AB - OBJECTIVE: Attention-deficit / Hyperactivity Disorder (ADHD) is a frequent mental disorder with onset in childhood and persistence into adulthood in a sizeable number of people. Despite a rather simple clinical definition, ADHD has many facets because of frequent co-morbid disorders and varying impact on psychosocial functioning. Thus, there is considerable heterogeneity in various domains. METHOD: A review of recent research findings in: i) selected domains of aetiology reflecting the role of genes, brain structures and functioning and the interplay of causal factors and ii) clinical heterogeneity in terms of co-morbidities, gender effects, courses and outcomes. RESULTS: Molecular genetic studies have identified a number of candidate genes which have a small effect on behavioural variation in ADHD. In the most recent Genome Scan Meta Analysis of seven ADHD linkage studies, genome-wide significant linkage was identified on chromosome 16. The volume of both the total brain and various regions including the prefrontal cortex, the caudate nucleus and the vermis of the cerebellum is smaller in ADHD. Functional MRI has documented a specific deficit of frontostriatal networks in ADHD. Integrative aetiological models have to take the interaction of gene and environment on various dysfunctions into account. Clinical heterogeneity results from frequent associations with various co-morbidities, the impact of the disorder on psychosocial functioning, and gender effects. Partly, these effects are evident also in the course and outcome of ADHD. CONCLUSION: ADHD is a chronic mental disorder with a complex aetiology. So far, various neurobiological factors have been identified that need to be studied further to better understand their interaction with environmental factors. The clinical presentation and the long term course of ADHD are manifold. PMID- 19807722 TI - Applied psychometrics in clinical psychiatry: the pharmacopsychometric triangle. AB - OBJECTIVE: To consider applied psychometrics in psychiatry as a discipline focusing on pharmacopsychology rather than psychopharmacology as illustrated by the pharmacopsychometric triangle. METHOD: The pharmacopsychological dimensions of clinically valid effects of drugs (antianxiety, antidepressive, antimanic, and antipsychotic), of clinically unwanted effects of these drugs, and the patients' own subjective perception of the balance between wanted and unwanted effects are analysed using rating scales assessed by modern psychometric tests (item response theory models) RESULTS: Symptom rating scales fulfilling the item response theory models have been shown to be psychometrically valid outcome scales as their total scores are sufficient statistics for demonstrating dose-response relationship within the various classes of antianxiety, antidepressive, antimanic or antipsychotic drugs. The total scores of side-effect rating scales are, however, not sufficient statistics, implying that each symptom has to be analysed individually. Self-rating scales with very few items appear to be sufficient statistics when measuring the patients' own perception of quality of life. CONCLUSION: Applied psychometrics in psychiatry have been found to cover a pharmacopsychometric triangle illustrating the measurements of wanted and unwanted effects of pharmacotherapeutic drugs as well as health-related quality of life. PMID- 19807723 TI - Effects of aerobic fitness on hypohydration-induced physiological strain and exercise impairment. AB - AIM: Hypohydration exacerbates cardiovascular and thermal strain and can impair exercise capacity in temperate and warm conditions. Yet, athletes often dehydrate in exercise, are hypervolaemic and have less cardiovascular sensitivity to acute hypervolaemia. We tested the hypothesis that trained individuals have less cardiovascular, thermoregulatory and performance affect of hypohydration during exercise. METHODS: After familiarization, six trained [VO(2 peak) = 64 (SD 8) mL kg(-1) min(-1)] and six untrained [O(2 peak) = 45 (4) mL kg(-1) min(-1)] males cycled 40 min at 70%O(2 peak) while euhydrated or hypohydrated by 1.5-2.0% body mass (crossover design), before a 40-min work trial with euhydration or ad libitum drinking (in Hypohydration trial), in temperate conditions (24.3 degrees C, RH 50%, v(a) = 4.5 m s(-1)). Baseline hydration was by complete or partial rehydration from exercise+heat stress the previous evening. RESULTS: During constant workload, heart rate and its drift were increased in Hypohydration compared with Euhydration for Untrained [drift: 33 (11) vs. 24 beats min(-1) h( 1) (10), 95% CI 5-11] but not Trained [14 (3) vs. 13 beats min(-1) h(-1) (3), CI 2 to 3; P = 0.01 vs. Untrained]. Similarly, rectal temperature drift was faster in Hypohydration for Untrained only [by 0.57 degrees C h(-1) (0.25); P = 0.03 vs. Trained], concomitant with their reduced sweat rate (P = 0.05) and its relation to plasma osmolality (P = 0.03). Performance power tended to be reduced for Untrained (-13%, CI -35 to 2) and Trained (-7%, CI: -16 to 1), without an effect of fitness (P = 0.38). CONCLUSION: Mild hypohydration exacerbated cardiovascular and thermoregulatory strain and tended to impair endurance performance, but aerobic fitness attenuated the physiological effects. PMID- 19807724 TI - Inaccurate endoscopy: a better explanation for placebo-associated endoscopic ulcers. PMID- 19807726 TI - Saccharomyces boulardii for treating acute gastroenteritis in children: updated meta-analysis of randomized controlled trials. PMID- 19807727 TI - Azathioprine plus ribavirin treatment and pancytopenia. PMID- 19807728 TI - Does acetyl salicylic acid (ASA) have a role in the prevention of venous thromboembolism? PMID- 19807729 TI - Generation and characterisation of Rhd and Rhag null mice. AB - Mouse Rhd* and Rhag* genes were targeted using insertional vectors; the resulting knockout mice, and double-knockout descendants, were analysed. Rhag glycoprotein deficiency entailed defective assembly of the erythroid Rh complex with complete loss of Rh and intercellular adhesion molecule 4 (ICAM-4), but not CD47, expression. Absence of the Rh protein induced a loss of ICAM-4, and only a moderate reduction of Rhag expression. Double knockout phenotype was similar to that of Rhag targeted mice. Rhd and Rhag deficient mice exhibited neither the equivalent of human Rh(null) haemolytic anaemia nor any clinical or cellular abnormalities. Rhd-/- and Rhag-/- erythrocytes showed decreased basal adhesion to an endothelial cell line resulting from defective ICAM-4 membrane expression. There was no difference in recovery from phenylhydrazine-induced haematopoietic stress for double knockout mice as compared to controls, suggesting that ICAM-4 might be dispensable during stress erythropoiesis. Ammonia and methylammonia transport in erythrocytes was severely impaired in Rhag-/- but only slightly in Rhd-/- animals that significantly expressed Rhag, supporting the view that RhAG and Rhag, but not Rh, may act as ammonium transporters in human and mouse erythrocytes. These knockout mice should prove useful for further dissecting the physiological roles of Rh and Rhag proteins in the red cell membrane. PMID- 19807730 TI - Multiplex ligation-dependent probe amplification screening of isolated increased HbF levels revealed three cases of novel rearrangements/deletions in the beta globin gene cluster. AB - Investigations of naturally occurring mutations, such as the deletional thalassaemias and hereditary persistence of fetal haemoglobins (HPFHs), have brought many insights into human globin switching, but limited data have been reported so far. We selected 15 individuals with elevated fetal haemoglobin (HbF) levels (>5%) from a previous screening of 27 006 Korean individuals and analysed dosage changes of the globin gene cluster using multiplex ligation-dependent probe amplification (MLPA). Dosage changes detected by the MLPA probes were followed up with gap-polymerase chain reaction and sequence analysis. Three subjects were found to have deletions in the globin gene cluster, including a beta-thalassaemia due to deletion of HBB (beta-globin gene), an HPFH due to deletions of HBD (delta-globin gene) and HBB, and an HPFH due to a novel HBG2 HBG1 fusion gene consisting of exons 1 and 2 of HBG2 ((G)gamma-globin gene) and exon 3 of HBG1 ((A)gamma-globin gene). The case with the HBG2-HBG1 fusion suggested the existence of another mechanism for the reactivation of HBG2 and HBG1. The IVS2 of HBG2 and HBG1might play a role in HbF regulation, and combinations of specific polymorphisms could influence the reactivation of these genes in adults. PMID- 19807731 TI - Gene silencing of MIR22 in acute lymphoblastic leukaemia involves histone modifications independent of promoter DNA methylation. AB - Aberrant epigenetic regulation has recently been implicated in the downregulation of tumour suppressor microRNAs (miRNAs). Histone modification and DNA methylation can have different roles in gene silencing in cancer. To investigate whether histone modifications would contribute to the dysregulation of miRNAs in acute lymphoblastic leukaemia (ALL), the effect of a histone deacetylase inhibitor, trichostatin A (TSA), on miRNA expression profile was analysed by microarray assay in a precursor B-cell ALL cell line NALM-6. A total of 10 miRNAs were downregulated and 31 were upregulated significantly following TSA treatment. Among TSA-upregulated miRNAs, MIR22 is an extronic miRNA and resides in the second exon of the non-coding transcript MGC14376. Upregulation of MIR22 transcription was found in both NALM-6 cells and primary human ALL malignant cells treated with TSA. Whereas a CpG island was identified within the promoter element of MIR22, no promoter DNA methylation was detected in these cells. In contrast, accumulation of the repressive histone marker H3K27 trimethylation (H3K27triM) was identified around the transcriptional start point of the gene, which was reduced by TSA treatment. Thus, accumulation of H3K27triM independent of promoter DNA methylation may be a novel epigenetic mechanism for MIR22 silencing in ALL. PMID- 19807732 TI - Thrombolysis in children. AB - Thrombosis in children may result in vascular occlusion and a potential loss of viability in skin, limbs or organs, or lead to the development of post thrombotic syndrome. Thrombolysis may prevent such complications, and the use of thrombolytics in children has increased in the last three decades. Despite this, optimum paediatric treatment regimens for these drugs remain uncertain, and accurate efficacy and safety profiles for thrombolytics are limited by the lack of controlled, prospective studies in well-defined patient populations. Restoration of vessel patency has often been achieved but the risk of intracranial and other major haemorrhage secondary to thrombolysis continues to be a significant concern for the clinician deciding on such treatment. This review summarises the use of thrombolytic treatment in children and describes outcome data and adverse events following treatment, as well as the limitations of published studies on paediatric thrombolysis. PMID- 19807733 TI - Active site ring-opening of a thiirane moiety and picomolar inhibition of gelatinases. AB - (+/-)-2-[(4-Phenoxyphenylsulfonyl)methyl]thiirane 1 is a potent and selective mechanism-based inhibitor of the gelatinase sub-class of the zinc-dependent matrix metalloproteinase family. Inhibitor 1 has excellent activity in in vivo models of gelatinase-dependent disease. We demonstrate that the mechanism of inhibition is a rate-limiting gelatinase-catalyzed thiolate generation via deprotonation adjacent to the thiirane, with concomitant thiirane opening. A corollary to this mechanism is the prediction that thiol-containing structures, related to thiirane-opened 1, will possess potent matrix metalloproteinase inhibitory activity. This prediction was validated by the synthesis of the product of this enzyme-catalyzed reaction on 1, which exhibited a remarkable K(i) of 530 pm against matrix metalloproteinase-2. Thiirane 1 acts as a caged thiol, unmasked selectively in the active sites of gelatinases. This mechanism is unprecedented in the substantial literature on inhibition of zinc-dependent hydrolases. PMID- 19807734 TI - Laser microdissection-based analysis of cytokine balance in the kidneys of patients with lupus nephritis. AB - To determine the cytokine balance in patients with lupus nephritis (LN), we analysed kidney-infiltrating T cells. Renal biopsy samples from 15 systemic lupus erythematosus (SLE) patients were used. In accordance with the classification of International Society of Nephrology/Renal Pathology Society, they were categorized into Class III, Class III+V (Class III-predominant group, n = 4), Class IV, Class IV+V (Class IV-predominant group, n = 7) and Class V (n = 4) groups. The single-cell samples of both the glomelular and interstitial infiltrating cells were captured by laser-microdissection. The glomerular and interstitial infiltrating T cells produced interleukin (IL)-2, IL-4, IL-10, IL-13 and IL-17 cytokines in the Class III-predominant, Class IV-predominant and Class V groups. Interferon-gamma was detected only in the glomeruli of the Class III predominant and Class V group samples. The expression level of IL-17 was correlated closely with clinical parameters such as haematuria, blood urea nitrogen level, SLE Disease Activity Index scores in both glomeruli and interstitium, urine protein level in glomeruli and serum creatinine and creatinine clearance levels in interstitium. This suggests that the glomerular infiltrating T cells might act as T helper type 1 (Th1), Th2 and Th17 cells while the interstitial infiltrating T cells, act as Th2 and Th17 cells in the Class III predominant and Class V groups. In contrast, both the glomerular and interstitial infiltrating T cells might act as Th2 and Th17 cells in the Class IV-predominant group. The cytokine balances may be dependent upon the classification of renal pathology, and IL-17 might play a critical role in SLE development. PMID- 19807735 TI - Identification and functional analysis of novel mutations of the CLCNKB gene in Chinese patients with classic Bartter syndrome. AB - Mutations in the gene CLCNKB encoding the ClC-Kb chloride channel causes classic Bartter syndrome, which is characterized by hypokalaemic metabolic alkalosis, renal salt loss, hyper-reninaemic hyperaldosteronism and normal blood pressure. We aimed to investigate the underlying mutations in CLCNKB in two Chinese patients with classic Bartter syndrome and then test the effect of the mutations on ClC-Kb chloride channel activity. Mutation analysis of CLCNKB was performed by polymerase chain reaction (PCR) direct sequencing. Expression of the wild-type and mutant ClC-Kb was heterologous in Xenopus laevis oocytes. We identified three novel CLCNKB gene mutations, including one homozygous missense mutation (R351W) in one patient and two compound heterozygous mutations (R30X and A210V) in the other. As determined by two-electrode voltage-clamp analysis of ClC-Kb channel activity, R30X abolished the current amplitude; A210V and R351W significantly reduced the current amplitude. A210V was almost as sensitive as the wild type to extracellular pH and calcium, whereas R351W removed extracellular calcium activation and markedly reduced alkaline pH activation of ClC-Kb. The three novel CLCNKB mutations we identified in two Chinese patients with classic Bartter syndrome have a role in altering the functional properties of ClC-Kb channels. PMID- 19807736 TI - CDKL5 truncation due to a t(X;2)(p22.1;p25.3) in a girl with X-linked infantile spasm syndrome. PMID- 19807737 TI - Novel CLN8 mutations confirm the clinical and ethnic diversity of late infantile neuronal ceroid lipofuscinosis. AB - The neuronal ceroid lipofuscinoses (NCLs) are a group of inherited lysosomal storage diseases and the prototype of childhood onset neurodegenerative disorders. To date, 10 NCL entities (CLN1-CLN10) are known and characterized by accumulation of autofluorescent storage material, age of onset and clinical symptoms. CLN8 was first identified as the causative gene for a late-onset form with progressive epilepsy and mental retardation in Finnish patients. In addition, CLN8 phenotypes were described in Turkish, Israeli and Italian patients with a more rapid progression of visual loss, epilepsy, ataxia and mental decline. Here, we report the first mutations in German (c.611G>T) and Pakistani (c.709G>A) patients. Our findings confirm previous assumptions that the CLN8 variant can occur in many ethnic groups. So far, large CLN gene deletions are only known for the CLN3 gene. Here, we also describe a novel, large CLN8 gene deletion c.544-2566_590del2613 in a Turkish family with a slightly more severe phenotype. Our data indicate that patients with clinical signs of late infantile NCL and characteristic ultrastructural inclusions should also be screened for CLN8 mutations independent of their ethnic origin. PMID- 19807738 TI - Genetic mutation in pontocerebellar hypoplasia. PMID- 19807739 TI - Two novel AIRE mutations in autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) among Indians. AB - Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a rare recessive disorder resulting from mutations in the autoimmune regulator (AIRE) gene. There is no information on AIRE mutations in Indians. In a cross-sectional study, nine patients (eight families), from four referral hospitals in India, were studied for AIRE mutations by direct sequencing. We screened for new mutations in 150 controls by allele-specific PCR. The patients had 1-7 known components of APECED. Three patients had unusual manifestations: presentation with type 1 diabetes; chronic sinusitis and otitis media; and facial dysmorphism. All patients carried homozygous, probably recessive, AIRE mutations. Two unrelated patients from a small in-bred community (Vanika Vaisya) in south India carried an unreported missense mutation, p.V80G, in the N-terminal caspase recruitment domain. Another unique mutation, p.C302X, resulting in a truncated protein with deletion of both zinc-finger domains, was detected in a patient from Gujarat. Neither mutation was detected in controls. Other mutations, previously described in Caucasians, were: 13 base pair deletion (p.C322fsX372) in 4 (38%), and Finn-major (p.R257X) and p.R139X (Sardinian) mutation in one subject each. In conclusion, in this first series of APECED in Indians, we detected AIRE mutations previously reported in Caucasians, as well as unique mutations. Of these, p.V80G is possibly an ancestral mutation in an in-bred community. PMID- 19807741 TI - Bilateral frontoparietal polymicrogyria (BFPP) syndrome secondary to a 16q12.1 q21 chromosome deletion involving GPR56 gene. PMID- 19807740 TI - Chromosome imbalances in syndromic hearing loss. AB - The cause of hearing impairment has not been elucidated in a large proportion of patients. We screened by 1-Mb array-based comparative genomic hybridization (aCGH) 29 individuals with syndromic hearing impairment whose clinical features were not typical of known disorders. Rare chromosomal copy number changes were detected in eight patients, four de novo imbalances and four inherited from a normal parent. The de novo alterations define candidate chromosome segments likely to harbor dosage-sensitive genes related to hearing impairment, namely 1q23.3-q25.2, 2q22q23, 6p25.3 and 11q13.2-q13.4. The rare imbalances also present in normal parents might be casually associated with hearing impairment, but its role as a predisposition gene remains a possibility. Our results show that syndromic deafness is frequently associated with chromosome microimbalances (14 27%), and the use of aCGH for defining disease etiology is recommended. PMID- 19807742 TI - Five novel germline function-impairing mutations of CYLD in Italian patients with multiple cylindromas. PMID- 19807743 TI - The ryanodine receptor type 1 gene variants in African American men with exertional rhabdomyolysis and malignant hyperthermia susceptibility. AB - It has been suggested that exertional rhabdomyolysis (ER) and malignant hyperthermia (MH) are related syndromes. We hypothesize that patients with unexplained ER harbor mutations in the ryanodine receptor gene type 1 (RYR1), a primary gene implicated in MH, and therefore ER patients are at increased risk for MH. Although there are reported cases of MH in individuals of African descent, there are no data available on molecular characterization of these patients. We analyzed RYR1 in six, unrelated African American men with unexplained ER, who were subsequently diagnosed as MH susceptible (MHS) by the Caffeine Halothane Contracture Test. Three novel and two variants, previously reported in Caucasian MHS subjects, were found in five studied patients. The novel variants were highly conserved amino acids and were absent among 230 control subjects of various ethnic backgrounds. These results emphasize the importance of performing muscle contracture testing and RYR1 mutation screening in patients with unexplained ER. The MHS-associated variant Ala1352Gly was identified as a polymorphism predominant in individuals of African descent. Our data underscore the need for investigating RYR1 across different ethnic groups and will contribute to interpretation of genetic screening results of individuals at risk for MH. PMID- 19807744 TI - Primary congenital glaucoma caused by the homozygous F261L CYP1B1 mutation and paternal isodisomy of chromosome 2. AB - Primary congenital glaucoma (PCG), a rare, severe and blinding disease, usually results from mutations in the CYP1B1 gene located in chromosome 2p22.2. Uniparental isodisomy (UPID) is also a rare condition in which a diploid offspring carries two identical copies of a single parental chromosome. By DNA sequence analysis, we found that a proband (female newborn) affected by PCG was homozygous for the null-allele F261L of the CYP1B1 gene. Her father was a heterozygous carrier for this mutation, and unexpectedly her mother carried only the G168D mutation in the heterozygous state. Segregation analysis of eight microsatellite markers which spanned the two arms of chromosome 2 was consistent with paternal isodisomy for this chromosome in the proband. To the best of our knowledge, this is the first reported case of UPID resulting in PCG and the fifth reported case of paternal UPID for chromosome 2. In addition, the absence of a clinical phenotype other than PCG confirms previous observations of there being no paternally imprinted genes in chromosome 2 that have major phenotypic effects. These results, along with previous reports, also suggest that UPID may play a relevant role in recessive diseases linked to chromosome 2. PMID- 19807745 TI - Flowmetry-based portal inflow manipulation for a small-for-size liver graft in a recipient with spontaneous splenorenal shunt. AB - We report a case of living donor liver transplantation using a small-for-size graft (SFSG) with graft to estimated standard liver volume of only 28% in a recipient with spontaneous splenorenal shunt and demonstrate the value of intraoperative ultrasonic flowmetry. Despite an SFSG, the graft was underperfused. This was recognized by flowmetry and was rectified by ligation of the splenorenal shunt. PMID- 19807746 TI - Selecting an optimal cutoff value for creatinine in the model for end-stage liver disease equation. AB - BACKGROUND: The model for end-stage liver disease (MELD) is used for organ allocation in liver transplantation. The maximal serum creatinine (Cr) level for MELD is set at 4.0 mg/dL; however, there was no outcome data to justify this strategy. METHODS: Ninety-two patients with cirrhosis with Cr level >4 mg/dL were selected from 1438 patients and compared with MELD score-matched controls for three-month and six-month mortality. RESULTS: At three months, patients with Cr level >4 mg/dL had a significantly higher mortality rate than the 184 controls with a lower Cr level (44.6% vs. 29.3%, p = 0.015). This trend was still significant at six months: the mortality rate was 62% in the index group vs. 45.1% in the control group (p = 0.011). The difference between the index and control groups was the smallest (2.5% at three months and 3.4% at six months) when Cr was up-scaled to 5.5 mg/dL. The predictive accuracy of the MELD was estimated by using area under receiver-operating characteristic (AUC) curve. Only the cutoff of 5.5 mg/dL at six months displayed a higher AUC (0.753). CONCLUSIONS: A cutoff at 5.5 mg/dL may be more appropriate for the MELD. The MELD for patients with cirrhosis with advanced renal insufficiency deserves re evaluation. PMID- 19807748 TI - The association between hand eczema and nickel allergy has weakened among young women in the general population following the Danish nickel regulation: results from two cross-sectional studies. AB - BACKGROUND: An association between nickel contact allergy and hand eczema has previously been demonstrated. In 1990, Denmark regulated the extent of nickel release in the ear-piercing process as well as nickel release from consumer products. OBJECTIVES: This study aimed to evaluate the effect of the Danish nickel regulation by comparing the prevalence of concomitant nickel allergy and hand eczema observed in two repeated cross-sectional studies performed in the same general population in Copenhagen. MATERIALS: In 1990 and 2006, 3881 18-69 year olds completed a postal questionnaire and were patch tested with nickel. Data were analysed by logistic regression analyses and associations were expressed as odds ratios (ORs) with 95% confidence intervals (CIs). RESULTS: The prevalence of concomitant nickel contact allergy and a history of hand eczema decreased among 18-35-year-old women from 9.0% in 1990 to 2.1% in 2006 (P < 0.01). The association between nickel contact allergy and a history of hand eczema decreased in this age group between 1990 (OR = 3.63; CI = 1.33-9.96) and 2006 (OR = 0.65; CI = 0.29-1.46). Among older women, no significant changes were observed in the association between nickel contact allergy and hand eczema. CONCLUSIONS: Regulatory control of nickel exposure may have reduced the effect of nickel on hand eczema in the young female population. PMID- 19807749 TI - The possible relevance of sex hormones on irritant and allergic responses: their importance for skin testing. AB - Dermatological responses are affected by the menstrual cycle phase in female patients, an unsurprising observation as oestrogen and progesterone affect the skin and immunological function, with oestrogen suppression of cellular immunity in particular. Exacerbation of dermatological symptoms is typically observed in either the latter phase of the menstrual cycle or during menstruation. The allergic response is diminished in the ovulatory phase and heightened in the progestinic phase. Definitive conclusions with regard to the effect of reproductive hormones on skin disorders have been somewhat hampered by a body of research that has employed diverse research parameters, such as dosage, testing sites, concentration, vehicle of irritant delivery, and method of assessment, however, individual patient sensitivity varies widely. Standardization of measurement techniques is necessary to provide reproducible results as much as individual patient variation and technique will allow. PMID- 19807750 TI - Reactions to resin-based dental materials in patients--type, time to onset, duration, and consequence of the reaction. AB - OBJECTIVE: The aim of the present study was to determine the types of side effects occurring and for how long they lasted in a group of patients with side effects assessed to be caused by resin-based materials. METHODS: A total of 618 reports were received by the Swedish National Register of Side-Effects to Dental Materials, among which 36 were on patients with reactions assessed to be caused by resin-based restorative materials. The group examined consisted of 25 women and 11 men, with a mean age of 47.8 +/- 15.6 years. A follow-up was done through a structured telephone interview. RESULTS: The majority of symptoms were intra oral or a combination of intra-oral and extra-oral symptoms that appeared within the first 24 hr after treatment. The most common adverse effects reported were skin problems, oral ulcers, and burning mouth. Within less than a week, the reactions had disappeared in 50% of the patients. CONCLUSION: Immediate reactions to resin-based materials were more prevalent than delayed allergic reactions, and the mechanism of the immediate reactions is probably non-allergic in most cases. There is a need for developing provocation tests to verify the association between the reaction and the material, and also to identify the offending component. PMID- 19807751 TI - Does allergic contact dermatitis from formaldehyde in clothes treated with durable-press chemical finishes exist in the USA? AB - Recent US studies have presented case series of patient with allergic contact dermatitis (ACD) allegedly caused by formaldehyde in clothes treated with durable press chemical finishes (DPCF), which are known formaldehyde releasers. However, the amounts of formaldehyde released by modern DPCF are thought to be well below the levels previously estimated to be able to elicit ACD. The objectives of this review are (i) to investigate whether clothes sold in the USA may contain enough free formaldehyde to elicit ACD in previously sensitized individuals and (ii) to assess the validity of US reports on ACD from formaldehyde in DPCF treated clothes. Literature was examined using various resources. The threshold level for formaldehyde in clothes that may cause ACD in sensitized individuals is unknown; we present data suggesting that levels < 200 ppm will be safe for most patients and that textiles will rarely contain higher amounts. All US studies presenting patients with ACD from formaldehyde in clothes had some weaknesses and in no report was the diagnosis proven beyond doubt. Currently, there is no definite proof that textile ACD from formaldehyde in DPCF in the USA exists. Future research should be directed at establishing the elicitation threshold and the amounts of formaldehyde present in textiles. PMID- 19807752 TI - A special issue devoted to filler complications. PMID- 19807753 TI - The nature of long-term fillers and the risk of complications. PMID- 19807754 TI - Lexicon for soft tissue implants. PMID- 19807756 TI - Complications after treatment with polyalkylimide. AB - BACKGROUND: Polyalkylimide is a nonresorbable, biocompatible polymeric filler that has been used for several years to treat soft tissue deficits. The literature has shown a minor complication rate. We noticed that complications typically appear several years after injection. OBJECTIVE: To evaluate the complications reported after treatment with polyalkylimide. METHODS AND MATERIALS: We describe a retrospective evaluation, reported by members and candidate members of the Dutch Society of Cosmetic Medicine, of complications after use of polyalkylimide. RESULTS: In total, 3,196 patients were treated, and 4,738 treatments were performed, from which 154 complications (patient complication rate 4.8%, treatment complication rate 3.3%) were reported. The most common complication was inflammation; other complications were hardening, migration, and accumulation of the product. In some patients, skin biopsy followed by histologic examination was performed. CONCLUSION: Treatments with polyalkylimide have been reported to give rise to complications years after treatment. Even though the study described is a retrospective evaluation, we consider an overall complication rate of 4.8%, the severity of the complications, and the difficulty in treating them too high a risk for a cosmetic treatment. The Dutch Society of Cosmetic Medicine advises against the use of polyalkylimide. PMID- 19807755 TI - Host tissue interaction, fate, and risks of degradable and nondegradable gel fillers. AB - BACKGROUND: A constantly increasing number of gel fillers for aesthetic and reconstructive purposes have been introduced during the last 20 years. Most of the new ones are modified versions of the original collagen and hyaluronic acid gels. They have been reconstructed, often by adding cross-bindings to the polymer in order to obtain a more dense molecular structure, which will prolong degradation and filling effect of the gel. Other gel fillers contain particles of organic (poly-lactic acid) or inorganic (calcium hydroxylapatite) material, which have been used in human tissue for other purposes (degradable suture material and bone cement, respectively). The permanent fillers (silicone oil and polyacrylamide gel) have been used for many years, silicone mainly in the US and polyacrylamide gel in most countries outside the US and Canada. OBJECTIVE: Complications occur, and they appear to be more frequent with particulated fillers, polyacrylamide gel and silicone oil. However, these complications differ in nature and depend on the filler type used. METHODS AND MATERIALS: This overview presents the different gel filler types, how they interact with host tissue, and what can go wrong. The results and conclusion are based on experimental and clinical observations coupled with a search of the literature. RESULTS AND CONCLUSION: Complications following homogenous hydrogels are caused by infection with bacteria, which have been inserted into the gel during injection. If not treated with relevant antibiotics (but instead steroids or large doses of NSAIDs) the bacteria form a biofilm, which gives rise to a low grade chronic infection that is resistant to antibiotics. Complications following particulated gels and silicone oil are not known, but bacteria in a biofilm and/or endotoxins released by these is a possibility which deserves further investigations, primarily by using the fluorescence in situ hybridization (FISH) technique. PMID- 19807757 TI - The spectrum of adverse reactions after treatment with injectable fillers in the glabellar region: results from the Injectable Filler Safety Study. AB - BACKGROUND: For the glabellar region, severe partly vascular adverse events have been reported after treatment with injectable fillers. METHODS AND MATERIALS: For this study, data from the Injectable Filler Safety Study, a German-based registry for those reactions, was analyzed to characterize adverse events seen in the glabellar region. Patients were analyzed descriptively. RESULTS: Forty of 139 registered patients reported adverse events in the glabellar region. All patients were female, with an average age of 52.3. Nineteen patients with adverse reactions to hydroxyethylmethacrylate (HEMA) and ethylmethacrylate (EMA) in a fixed combination with hyaluronic acid (HA) and 10 patients with adverse reactions to different hyaluronic acid products were reported; five patients reacted to poly-L-lactic acid (PLA). The most common adverse reactions to HEMA/EMA in HA and PLA were nodules and hardening. In HA-treated patients, erythema and inflammation, swelling, and pain were most frequent. The adverse reactions to HEMA/EMA in HA were severe in 50% of the patients. Severe adverse reactions were found to a lesser extent in patients treated with HA and PLA. Potential vascular complications were documented in only two patients. CONCLUSION: Adverse reactions seen in the glabella are overwhelmingly product associated and to a lesser extent location associated. Vascular complications with necrosis and ulceration were rare. PMID- 19807758 TI - The risk of alar necrosis associated with dermal filler injection. AB - BACKGROUND: Injection of dermal fillers is one of the most commonly performed cosmetic procedures. Serious complications from fillers are rare but potentially devastating to patients and physicians. Skin necrosis, such as nasal alar necrosis, is one of the most feared serious complications of dermal fillers, but there is a paucity of literature on the incidence of such events, as well as potential treatment options. METHODS: We present a review of the literature and three cases of nasal alar necrosis after dermal filler injection. CONCLUSION: Nasal alar necrosis associated with dermal filler injection is a rare event. Proper technique and recognition of risk factors may reduce the incidence of this complication. Physicians should be aware of early intervention and treatment options should impending necrosis become apparent. PMID- 19807759 TI - Six-year experience using 1,000-centistoke silicone oil in 916 patients for soft tissue augmentation in a private practice setting. AB - BACKGROUND: There are no previously published reports focusing exclusively on the use of 1,000-centistoke purified polydimethylsiloxane (PDMS-1000) for cosmetic soft-tissue augmentation. OBJECTIVE: To provide clinical experience with its cosmetic use, solely and in conjunction with other nonpermanent fillers, in a private practice setting. METHODS AND MATERIALS: A retrospective chart review was conducted for patients treated by the author over 6 years, beginning in 2003. Treatments were tabulated according to facial region and arbitrarily designated as rhytides, acne scars, lips, infraorbital, nasolabial, and general contour. Therefore, up to six treatments were possible with any visit. Concomitant treatment with nonpermanent fillers, as well as any significant adverse events, was noted as well. RESULTS: Nine hundred sixteen patients were treated (816 (89%) female, 100 (11%) male). There were 5,246 treatments over 3,307 visits, with an average of 3.5 visits per patient and 1.6 treatments per visit. Adverse events were limited to overcorrection in 11 patients (1%). Of the 916 patients, 257 (28%) were also treated with other (nonpermanent) fillers without incident. CONCLUSION: Over the 6-year period, PDMS-1000 was found to be effective and safe in the cosmetic practice setting. Other (nonpermanent) fillers were also used without incident. PMID- 19807760 TI - Safety of nonanimal stabilized hyaluronic acid dermal fillers in patients with skin of color: a randomized, evaluator-blinded comparative trial. AB - BACKGROUND: Nonanimal stabilized hyaluronic acid (NASHA) dermal fillers have been used in the United States since 2003 for the treatment of moderate to severe facial wrinkles and folds, such as nasolabial folds. Previous trials with NASHA dermal fillers have included small numbers of patients with pigmented skin. OBJECTIVES: This randomized, evaluator-blinded, split-face trial compared the safety and efficacy of two variable-particle NASHA fillers in the correction of nasolabial folds in patients with Fitzpatrick skin types IV, V, and VI. METHODS AND MATERIALS: One hundred fifty patients (predominantly African American) were enrolled at 10 sites. Patients received one treatment (with an optional touch-up treatment at week 2) with small- and large-particle NASHA gel randomized to the left or right side of the face. Safety was evaluated through patient diaries for the first 2 weeks and physician assessments at 3 days and 2, 6, 12, and 24 weeks after treatment for adverse events (AEs), skin pigmentation changes, and keloid formation. RESULTS: All related AEs were mild or moderate and of limited duration. They included bruising, tenderness, edema, redness, itching, pain, and changes in pigmentation. The incidence of AEs was not different between the 2 preparations, and no patient developed a keloid. There were 3 reported mass formations, 2 of which were infectious in nature. CONCLUSION: In patients with skin of color, NASHA dermal fillers are safe for the correction of moderate to severe facial folds and wrinkles, with no immunogenicity or keloid formation and only mild to moderate AEs occurring around the injection site. PMID- 19807761 TI - Clinical experience with filler complications. PMID- 19807762 TI - Recommendations and treatment options for nodules and other filler complications. PMID- 19807763 TI - Treatment of injectable soft tissue filler complications. AB - BACKGROUND: Increasing use of dermal fillers, especially with the U.S. introduction of long-term and permanent fillers, is expected to increase the number of complications seen after treatment with these materials. OBJECTIVES: To provide physicians with an overview for treatment of dermal filler complications. RESULTS: Complications of dermal filler use can be treated successfully based on a firm understanding of the mechanisms and limitations of filler action and wound healing. Complications can be classified as immediate, early, or delayed onset; available treatment options are presented. CONCLUSION: Short- and long-term dermal fillers are increasingly becoming popular treatment options for signs of facial aging. Complications, although not common, can be treated effectively and excellent outcomes achieved. PMID- 19807764 TI - Electron microscopic documentation of late changes in permanent fillers and clinical management of granulomas in affected patients. AB - BACKGROUND: The manufacturers of permanent injectable fillers claim that their products are widely inert, biocompatible, atoxic, and nonimmunogenic. There are polymer gels without microparticles on the market and combination products that use collagen suspension or a hyaluronic acid gel as a vector to which polymer microspheres or polygonal particles are added. The filling effect of the polymer gels is based on the volume injected and, for the combination gels, partly on the volume injected and partly on the intended host foreign-body reaction to the microparticles. Foreign body reactions that are seen as inflammatory, sometimes disfiguring, nodules may develop years later at the injection sites. OBJECTIVES: Permanent fillers differ with respect to composition and chemical and biological characteristics. There have been reports that intend to explain how host tissue reacts with different permanent fillers and how adverse reactions differ depending on the filler used. The changes that some of the permanent fillers undergo during years of residence in human tissue have not been included in this discussion. These structural changes may be one of the reasons why adverse reactions to permanent fillers occur clinically with a delay of several years. METHODS: In a series of 10 patients who had been injected with a permanent filler of hydroxymethylmethacrylate and ethylmethacrylate (40%) in hyaluronic acid gel (60%) and had developed adverse reactions with inflammatory nodules after variable time elapsed, biopsies could be obtained for histologic and electron microscopic examinations. RESULTS: After 2 years in all specimens, changes of degradation of the filler material could be detected. Bacteria were not found in any of the specimen. In 40% of the particles, the size of the particles did not correspond to the size declared by the manufacturer (45-65 microm) and was smaller, thus being more susceptible to phagocytosis. CONCLUSIONS: Inflammatory nodules due to adverse reactions to permanent fillers containing microparticles with a hydrophobic surface were treated with good results with a regimen of allopurinol and intralesional injections with a mixture of fluorouracil and low dose triamcinolon. PMID- 19807765 TI - Advanced laser techniques for filler-induced complications. AB - BACKGROUND: The increasing use of injectable fillers has been increasing the occurrence of disfiguring anaerobic infection or granulomas. This study presents two types of laser-assisted evacuation of filler material and inflammatory and necrotic tissue that were used to treat disfiguring facial nodules after different types of gel fillers. MATERIALS AND METHODS: Infectious lesions after hydrogels were drained using a lithium triborate laser at 532 nm, with subsequent removal of infected gel and pus (laser assisted evacuation). Granuloma after gels containing microparticles were treated using an 808-nm diode laser using intralesional laser technique. The latter melted and liquefied the organic and synthetic components of the granulomas, facilitating subsequent evacuation. Both lasers had an easily controllable thin laser beam, which enabled the physician to control tissue damage and minimize discomfort and pain. RESULTS: All 20 patients experienced reduction or complete resolution, the latter increasing with repeated treatments. CONCLUSION: Laser-assisted treatment offers a successful solution for patients who have been suffering from disfiguring nodules from injected fillers often for many years. The procedure broadens the range of treatment options in cases of untoward reactions to fillers, in line with surgical removal but with lower morbidity and less cosmetic disfigurement. PMID- 19807766 TI - Granulomatous foreign body reaction to hyaluronic acid: report of a case after melolabial fold augmentation and review of management. PMID- 19807767 TI - Foreign body reaction to hyaluronic acid filler injection: in search of an etiology. PMID- 19807768 TI - Hand function in relation to brain lesions and corticomotor-projection pattern in children with unilateral cerebral palsy. AB - AIM: To investigate relationships between hand function, brain lesions, and corticomotor projections in children with unilateral cerebral palsy (CP). METHOD: The study included 17 children (nine males, eight females; mean age 11.4 [SD 2.4] range 7-16 y), with unilateral CP at Gross Motor Function Classification System level I and Manual Ability Classification System level I or II. Hand function was assessed with the Box and Blocks test and Assisting Hand Assessment (AHA). Conventional structural magnetic resonance images were assessed visually for type, location, and extent of brain lesions. Single-pulse transcranial magnetic stimulation (TMS) provided information on organization of corticomotor projections. RESULTS: The most favourable hand function was seen in children who had white-matter damage of immaturity with mild white-matter loss and contralateral motor projections. Children with ipsilateral projections had the most impaired function. Nevertheless, in this subgroup a range of ability was seen (AHA 29-59%). Motor-projection patterns appeared to be influenced by lesion extent and location, but not by lesion type. INTERPRETATION: Combining information from structural magnetic resonance images and TMS can improve prediction of hand function. A wide variation in hand function was seen within all motor-projection patterns. Although the most impaired hand function was seen in the ipsilateral motor-projection group, some children in this group had fairly good ability. Such information is important for treatment planning. PMID- 19807769 TI - Genetic basis for acute necrotizing encephalopathy of childhood. PMID- 19807770 TI - Memory impairment in children with language impairment. AB - AIM: The aim of this study was to assess whether any memory impairment co occurring with language impairment is global, affecting both verbal and visual domains, or domain specific. METHOD: Visual and verbal memory, learning, and processing speed were assessed in children aged 6 years to 16 years 11 months (mean 9 y 9 m, SD 2 y 6 mo) with current, resolved, and no language impairment using the Wide Range Assessment of Memory and Learning (WRAML), a standardized memory and learning test for children, and the Children's Test of Non-Word Repetition (CNRep), a test of phonological short-term memory. Fifty-one children (38 males, 13 females) with current speech and language impairment from 49 families were compared with 13 siblings (11 males, 2 females) with a past history of language impairment and 26 (15 males, 11 females) who had never had language impairment. RESULTS: Children with current language impairment showed impairment in all verbal memory measures compared with children who had never had language impairment, and these impairments were still evident in children with a past history of learning impairment. Visual memory and learning were not impaired compared with children who had never had language impairment. The severity of verbal memory impairment correlated with the degree of language impairment. INTERPRETATION: We concluded that in language impairment there is domain specificity of memory impairment affecting verbal processing. PMID- 19807771 TI - Current and future uses of the Gross Motor Function Classification System: the need to take account of other factors to explain functional outcomes. PMID- 19807773 TI - Consumer-resource theory predicts dynamic transitions between outcomes of interspecific interactions. AB - Interactions between two populations are often defined by their interaction outcomes; that is, the positive, neutral, or negative effects of species on one another. Yet, signs of outcomes are not absolute, but vary with the biotic and abiotic contexts of interactions. Here, we develop a general theory for transitions between outcomes based on consumer-resource (C-R) interactions in which one or both species exploit the other as a resource. Simple models of C-R interactions revealed multiple equilibria, including one for species coexistence and others for extinction of one or both species, indicating that species' densities alone could determine the fate of interactions. All possible outcomes [(+ +), (+ -), (--), (+ 0), (- 0), (0 0)] of species coexistence emerged merely through changes in parameter values of C-R interactions, indicating that variation in C-R interactions resulting from biotic and abiotic conditions could determine shifts in outcomes. These results suggest that C-R interactions can provide a broad mechanism for understanding context- and density-dependent transitions between interaction outcomes. PMID- 19807774 TI - Linking dispersal, immigration and scale in the neutral theory of biodiversity. AB - In the classic spatially implicit formulation of Hubbell's neutral theory of biodiversity a local community receives immigrants from a metacommunity operating on a relatively slow timescale, and dispersal into the local community is governed by an immigration parameter m. A current problem with neutral theory is that m lacks a clear biological interpretation. Here, we derive analytical expressions that relate the immigration parameter m to the geometry of the plot defining the local community and the parameters of a dispersal kernel. Our results facilitate more rigorous and extensive tests of the neutral theory: we conduct a test of neutral theory by comparing estimates of m derived from fits to empirical species abundance distributions to those derived from dispersal kernels and find acceptable correspondence; and we generate a new prediction of neutral theory by investigating how the shapes of species abundance distributions change theoretically as the spatial scale of observation changes. We also discuss how our main analytical results can be used to assess the error in the mean-field approximations associated with spatially implicit formulations of neutral theory. PMID- 19807775 TI - Dynamic marine protected areas can improve the resilience of coral reef systems. AB - Marine Protected Areas are usually static, permanently closed areas. There are, however, both social and ecological reasons to adopt dynamic closures, where reserves move through time. Using a general theoretical framework, we investigate whether dynamic closures can improve the mean biomass of herbivorous fishes on reef systems, thereby enhancing resilience to undesirable phase-shifts. At current levels of reservation (10-30%), moving protection between all reefs in a system is unlikely to improve herbivore biomass, but can lead to a more even distribution of biomass. However, if protected areas are rotated among an appropriate subset of the entire reef system (e.g. rotating 10 protected areas between only 20 reefs in a 100 reef system), dynamic closures always lead to increased mean herbivore biomass. The management strategy that will achieve the highest mean herbivore biomass depends on both the trajectories and rates of population recovery and decline. Given the current large-scale threats to coral reefs, the ability of dynamic marine protected areas to achieve conservation goals deserves more attention. PMID- 19807776 TI - Parental involvement in paediatric cancer treatment decisions. AB - This study investigated parents' information needs and involvement in decision making processes affecting the care of children diagnosed with cancer. Interviews and questionnaires were used to assess parental satisfaction in 50 mothers and 16 fathers responsible for 58 children in an English Paediatric Oncology Unit. Parents reported that doctors contributed almost twice as much to the decision making process as they did, but parental satisfaction was positively correlated with the amount of information provided when giving informed consent. Satisfaction about their involvement in this process relied heavily upon the level of support received from others. Parents consenting to their child's involvement in non-randomised trials perceived themselves to be under greater pressure from others during the decision-making process while those whose children were further along the treatment trajectory were more uncertain about decisions previously made. Findings indicate that the accessibility, support, information and degree of control afforded to parents by healthcare professionals impacts upon their satisfaction with both the decision-making process and their confidence in the decisions thus made. Information and support tailored to parents' specific needs may therefore enhance satisfaction with clinical decision making and reassure parents about decisions made in the long-term interest of their child's health. PMID- 19807777 TI - Molecular dissection of bacterial acrylate catabolism--unexpected links with dimethylsulfoniopropionate catabolism and dimethyl sulfide production. AB - A bacterium in the genus Halomonas that grew on dimethylsulfoniopropionate (DMSP) or acrylate as sole carbon sources and that liberated the climate-changing gas dimethyl sulfide in media containing DMSP was obtained from the phylloplane of the macroalga Ulva. We identified a cluster that contains genes specifically involved in DMSP catabolism (dddD, dddT) or in degrading acrylate (acuN, acuK) or that are required to break down both substrates (dddC, dddA). Using NMR and HPLC analyses to trace 13C- or 14C-labelled acrylate and DMSP in strains of Escherichia coli with various combinations of cloned ddd and/or acu genes, we deduced that DMSP is imported by the BCCT-type transporter DddT, then converted by DddD to 3-OH-propionate (3HP), liberating dimethyl sulfide in the process. As DddD is a predicted acyl CoA transferase, there may be an earlier, unidentified catabolite of DMSP. Acrylate is also converted to 3HP, via a CoA transferase (AcuN) and a hydratase (AcuK). The 3HP is predicted to be catabolized by an alcohol dehydrogenase, DddA, to malonate semialdehyde, thence by an aldehyde dehydrogenase, DddC, to acyl CoA plus CO2. The regulation of the ddd and acu genes is unusual, as a catabolite, 3HP, was a co-inducer of their transcription. This first description of genes involved in acrylate catabolism in any organism shows that the relationship between the catabolic pathways of acrylate and DMSP differs from that which had been suggested in other bacteria. PMID- 19807778 TI - Shifts between Nitrospira- and Nitrobacter-like nitrite oxidizers underlie the response of soil potential nitrite oxidation to changes in tillage practices. AB - Despite their role in soil functioning, the ecology of nitrite-oxidizing bacteria, NOB, and their response to disturbances such as those generated by agricultural practices are scarcely known. Over the course of 17 months, we surveyed the potential nitrite oxidation, PNO, the abundance of the Nitrobacter- and Nitrospira-like NOB (by quantitative PCR) and the community structure of the Nitrobacter-like NOB (by PCR-DGGE and cloning-sequencing targeting the nxrA gene) in soils for four treatments: after establishment of tillage on a previously no tillage system, after cessation of tillage on a previously tillage system, and on control tillage and no-tillage systems. Key soil variables (moisture, organic carbon content and gross mineralization--i.e. ammonification--measured by the 15N dilution technique) were also surveyed. PNO was always higher for the no-tillage than tillage treatments. Establishment of tillage led to a strong and rapid decrease in PNO whereas cessation of tillage did not change PNO even after 17 months. PNO was strongly and positively correlated to the abundance of Nitrobacter-like NOB and was also strongly related to gross mineralization, a proxy of N-availability; in contrast, PNO was weakly and negatively correlated to the abundance of Nitrospira-like NOB. Selection of a dominant population was observed under no-tillage, and PNO was loosely correlated to the community structure of Nitrobacter-like NOB. Our results demonstrate that Nitrobacter-like NOB are the key functional players within the NOB community in soils with high N availability and high activity level, and that changes in PNO are due to shifts between Nitrospira-like and Nitrobacter-like NOB and to a weaker extent by shifts of populations within Nitrobacter-like NOB. PMID- 19807780 TI - Diversity of human colonic butyrate-producing bacteria revealed by analysis of the butyryl-CoA:acetate CoA-transferase gene. AB - Butyrate-producing bacteria play an important role in the human colon, supplying energy to the gut epithelium and regulating host cell responses. In order to explore the diversity and culturability of this functional group, we designed degenerate primers to amplify butyryl-CoA:acetate CoA-transferase sequences from faecal samples provided by 10 healthy volunteers. Eighty-eight per cent of amplified sequences showed >98% DNA sequence identity to CoA-transferases from cultured butyrate-producing bacteria, and these fell into 12 operational taxonomic units (OTUs). The four most prevalent OTUs corresponded to Eubacterium rectale, Roseburia faecis, Eubacterium hallii and an unnamed cultured species SS2/1. The remaining 12% of sequences, however, belonged to 20 OTUs that are assumed to come from uncultured butyrate-producing strains. Samples taken after ingestion of inulin showed significant (P=0.019) increases in Faecalibacterium prausnitzii. Because several of the dominant butyrate producers differ in their DNA % G+C content, analysis of thermal melt curves obtained for PCR amplicons of the butyryl-CoA:acetate CoA-transferase gene provides a convenient and rapid qualitative assessment of the major butyrate producing groups present in a given sample. This type of analysis therefore provides an excellent source of information on functionally important groups within the colonic microbial community. PMID- 19807779 TI - Characterization of marine isoprene-degrading communities. AB - Isoprene is a volatile and climate-altering hydrocarbon with an atmospheric concentration similar to that of methane. It is well established that marine algae produce isoprene; however, until now there was no specific information about marine isoprene sinks. Here we demonstrate isoprene consumption in samples from temperate and tropical marine and coastal environments, and furthermore show that the most rapid degradation of isoprene coincides with the highest rates of isoprene production in estuarine sediments. Isoprene-degrading enrichment cultures, analysed by denaturing gradient gel electrophoresis and 454 pyrosequencing of the 16S rRNA gene and by culturing, were generally dominated by Actinobacteria, but included other groups such as Alphaproteobacteria and Bacteroidetes, previously not known to degrade isoprene. In contrast to specialist methane-oxidizing bacteria, cultivated isoprene degraders were nutritionally versatile, and nearly all of them were able to use n-alkanes as a source of carbon and energy. We therefore tested and showed that the ubiquitous marine hydrocarbon-degrader, Alcanivorax borkumensis, could also degrade isoprene. A mixture of the isolates consumed isoprene emitted from algal cultures, confirming that isoprene can be metabolized at low, environmentally relevant concentrations, and suggesting that, in the absence of spilled petroleum hydrocarbons, algal production of isoprene could maintain viable populations of hydrocarbon-degrading microbes. This discovery of a missing marine sink for isoprene is the first step in obtaining more robust predictions of its flux, and suggests that algal-derived isoprene provides an additional source of carbon for diverse microbes in the oceans. PMID- 19807781 TI - Improvement of a dendritic cell-based tumour vaccine by an influenza virus. AB - BACKGROUND: Induction of cytotoxic T cells by dendritic cells (DCs) is a promising approach to tumour-immunotherapy. A standardized effective preparation of DCs remains a challenge for clinical application. MATERIAL AND METHODS: We assessed whether influenza A partial NS1 deletion (NS1-124) - or complete NS1 deletion (delNS1) vaccine viruses can be employed to enhance monocyte-derived dendritic cell (MODC)-based T-cell stimulation directed against malignant cells in vitro. RESULTS: Infection of cultures containing human MODCs and CD3(+) T cells with NS1 deletion viruses led to an increased induction of type I interferons and IL-6 compared with infection with wild-type virus. This correlated with the fact that infection of MODCs with NS1 deletion viruses but not with wild type virus led to stimulation of a cytotoxic T-cell (CTL) response against the Panc-1 cells, which were used as cell lysate to prime the MODCs. Moreover, stimulation of MODCs with Panc-1 tumour cell lysate obtained via lysis with the complete deletion virus delNS1, but not with the partial NS1 deletion virus also enhanced the CTL response against the tumour cells. Induction of function CTL response in those assays correlated with an increased proliferation of CD8(+) T cells. CONCLUSIONS: The pro-inflammatory capacity of influenza NS1 deletion vaccine viruses could serve as an adjuvant-like agent to improve preparations of MODC-based anti-cancer vaccines. The complete NS1 deletion virus appears to be more potent as adjuvant when used for production of tumour lysates. PMID- 19807782 TI - Age determines vascular reactivity as measured by optical rhinometry. AB - BACKGROUND: Endonasal blood volume depends on autonomous nervous system activation and ultimately vascular reactivity. It can be measured by optical rhinometry, a simple- to-use, subject-compliance independent method. We investigated endonasal perfusion following postural change to determine predictors of vascular response. MATERIALS AND METHODS: Optical rhinometry was performed in 52 healthy subjects (25 women) with an age range of 21-80 years. Subjects were placed in supine position after 5 min of standing. Optical density (OD) as marker of endonasal blood volume was continuously measured for 30 min after postural change. In addition to age, data on other risk factors for impaired vascular reactivity, such as gender, smoking, level of physical activity, resting heart rate and blood pressure were obtained. To evaluate raw data curves, integrals over defined time-periods were calculated. RESULTS: The OD initially increased following the intervention (t(max) at 207 +/- 12 s, mean +/- SEM). Thereafter, a gradual return to baseline levels was observed in young but not in elderly subjects. In multivariate models, age was the most influential parameter for vascular response followed by body mass index. The defined integrals as indicators of curve structure were all significantly influenced by age (P < 0.01 for each interval). Other measured risk factors had no effect on curve characteristics. CONCLUSIONS: Age is an important predictor of endonasal perfusion. Evaluation of optical rhinometry needs to take age-dependency of results into account. Studies in patients with cardiovascular disease are needed to show if optical rhinometry can provide information about vascular responsiveness as a predictor of cardiovascular morbidity. PMID- 19807783 TI - Spontaneous and bleomycin-induced chromosome damage in non cancer thyroid patients. AB - BACKGROUND: Presence of chromosome damage in lymphocytes of patients affected by several diseases, including cancer, was detected by the micronucleus (MN) assay. Individual susceptibility to DNA damage, considered as a risk factor for cancer, can be also evaluated using the bleomycin (BLM) sensitivity test. MATERIALS AND METHODS: We aimed to evaluate spontaneous or BLM-induced MN frequencies in autoimmune (AI, n = 19) and non autoimmune (NAI, n = 11) thyroid patients, not receiving (131)I radiometabolic therapy with respect to a control group of 18 healthy subjects. According to thyroid function, patients were also divided into hypothyroid (n = 10), euthyroid (n = 13) or hyperthyroid (n = 7) subjects. RESULTS: Spontaneous MN frequencies of AI and NAI patients did not differ from those of controls. Hypothyroid patients had more elevated MN basal levels (9.00 + or - 1.71 per thousand) than hyperthyroid (3.75 + or - 1.17 per thousand, P < 0.05) and euthyroid (5.38 + or - 0.97 per thousand, P < 0.01) patients or healthy subjects (4.17 + or - 0.63 per thousand, P < 0.01). In particular, the hypothyroid AI group showed the highest value (9.79 + or - 2.26 per thousand, P < 0.01). All thyroid patients responded differently to BLM than controls (39.90 + or - 2.48 per thousand vs. 31.08 + or - 2.51 per thousand, P = 0.0377). The NAI group had BLM-induced MN levels (45.00 + or - 2.56 per thousand) significantly higher (P = 0.0215) than AI patients (36.95 + or - 3.49 per thousand) or healthy subjects (31.08 + or - 2.51 per thousand). No significant difference was seen when patients were stratified according to autoimmunity. CONCLUSIONS: We report that hypothyroid patients exhibit a moderate increase in the level of spontaneous genome damage, and that AI thyroid patients resulted to be less sensitive than NAI patients to the mutagen sensitivity test. In prospective, it may be of interest to reinvestigate hypothyroid patients when correction of their dysfunction is achieved. PMID- 19807784 TI - Fat distribution influences the cardio-metabolic profile in a clinically healthy European population. AB - BACKGROUND AND AIMS: Waist and hip circumferences are largely influenced by Fat Mass and several other determinants. To evaluate the specific effects of a preferential fat distribution, we corrected the waist and hip circumferences for all their determinants. We then examined the association between fat distribution and several cardio-metabolic parameters in a clinically healthy population. SUBJECTS AND METHODS: In a subgroup of 625 females (F) and 490 males (M) from the RISC (Relationship between Insulin Sensitivity and Cardiovascular Disease) study, we evaluated insulin sensitivity by hyperinsulinaemic euglycaemic clamp and intima-media thickness (IMT) of the common (CCA) and internal (ICA) carotid artery by ultrasound imaging. Waist and hip circumferences were adjusted for age, height, fat and fat-free mass; in males, waist was also adjusted by hip and vice versa. RESULTS: Both F and M with enlarged waist showed significantly increased plasma insulin, C-peptide, total cholesterol, non-high density lipoprotein cholesterol, low density lipoprotein cholesterol and triglycerides, when compared with subjects with a smaller waist circumference. Males also showed lower glucose uptake and higher heart rate and ICA-IMT. A larger hip in both females and males was linked to a significantly greater inhibition of free fatty acids during the clamp test. CONCLUSION: Adjustment of waist circumference for its determinants permits the detection of early impairment of cardiovascular function and of glucose and lipid metabolism in a clinically healthy population, in particular in normal body weight subjects. Enlarged hip adjusted values are associated with greater insulin sensitivity. PMID- 19807785 TI - Antegrade flow and peripheral resistance determine the level of endogenous arteriogenesis in patients with superficial femoral artery occlusion. AB - OBJECTIVES: Occlusion in a limb artery leads to impaired blood supply and ischaemia. Collateral artery growth (arteriogenesis) is one of the most effective natural response mechanisms to compensate this pathologic situation. However, it is unknown if clinically important features, like poor run-off, have an impact on compensatory vessel growth. METHODS: Study population of this retrospective study consisted of 70 patients who suffered from lower limb ischaemia and underwent bypass surgery because of an occlusion of the superficial femoral artery. Clinical data were collected and pre- and postoperative angiograms were reviewed. Number of collateral vessels bypassing the occluded segment was counted. Features of inflow and outflow vessels were recorded. RESULTS: The mean number of collaterals was 13 + or - 0.5 per patient. In univariate analysis, short daily walking distance, chronic critical leg ischaemia, low ankle brachial index, low number of patent calf arteries and stenosed inflow arteries predicted low number of collateral arteries. In the multivariate analysis, only the quality of inflow and the number of patent calf vessels demonstrated an independent association (P < 0.05) with the number of collaterals. CONCLUSIONS: Ankle-brachial index, grade of symptoms and daily walking capacity could be used to predict collateral density. Importantly, a good antegrade flow and peripheral runoff seem to have a significant effect on collateral density, implying an impact on the activation of arteriogenesis. PMID- 19807787 TI - Deciphering the Leishmania exoproteome: what we know and what we can learn. AB - Parasitic protozoa of the genus Leishmania are the causative agents of leishmaniasis. Survival and transmission of these parasites in their different hosts require membrane-bound or extracellular factors to interact with and modify their host environments. Over the last decade, several approaches have been applied to study all the extracellular proteins exported by an organism at a particular time or stage in its life cycle and under defined conditions, collectively termed the secretome or the exoproteome. In this review, we focus on emerging data shedding light on the secretion mechanisms involved in the production of the Leishmania exoproteome. We also describe other methodologies currently available that could be used to analyse the Leishmania exoproteome. Understanding the complexity of the Leishmania exoproteome is a key component to elucidating the mechanisms used by these parasites for exporting proteins to the extracellular space during its life cycle. Given the importance of extracellular factors, a detailed knowledge of the Leishmania exoproteome may provide novel targets for rational drug design and/or a source of antigens for vaccine development. PMID- 19807786 TI - Oral administration of Lactococcus lactis expressing Helicobacter pylori Cag7 ct383 protein induces systemic anti-Cag7 immune response in mice. AB - To express the 3'-region (1152 bp) of the cag7 gene of Helicobacter pylori 51 strain, encoding the C-terminal 383 amino acid (ct383 aa) region of Cag7 protein that is known to cover the needle region of T4SS, in a live delivery vehicle Lactococcus lactis, the cag7-ct383 gene was amplified by PCR. DNA sequence analysis revealed that the amino acid sequence of Cag7-ct383 of H. pylori 51 shared 98.4% and 97.4% identity with H. pylori 26695 and J99, respectively. Intramuscular injection of the GST-Cag7-ct383 fusion protein into a rat could raise the anti-Cag7 antibody, indicating the immunogenicity of the Cag7-ct383 protein. When the cag7-ct383 gene was cloned in Escherichia coli-L. lactis shuttle vector (pMG36e) and transformed into L. lactis, the transformant could produce the Cag7-ct383 protein, as evidenced by Western blot analysis. The Cag7 ct383 protein level in the L. lactis transformant reached a maximum at the early stationary phase without extracellular secretion. The oral administration of the L. lactis transformant into mice generated anti-Cag7 antibody in serum in five of five mice. These results suggest that L. lactis transformant expressing Cag7 ct383 protein may be applicable as an oral vaccine to induce mucosal and systemic immunity to H. pylori. PMID- 19807788 TI - Complementary retrieval of 16S rRNA gene sequences using broad-range primers with inosine at the 3'-terminus: implications for the study of microbial diversity. AB - We evaluated the impact of the base analogue inosine substituted at the 3' terminus of broad-range 16S rRNA gene primers on the recovery of microbial diversity using terminal restriction fragment length polymorphism and clonal analysis. Oral plaque biofilms from 10 individuals were tested with modified and unmodified primer pairs. Besides a core overlap of shared terminal restriction fragments (T-RFs), each primer system provided unique information on the occurrence of T-RFs, with a higher number generally displayed with inosine primers. All clones sequenced were at least 99% identical to publicly available full-length sequences. Analysis of the corresponding primer-binding sites showed that most sequence types were 100% complementary to the unmodified primers so that the characteristic of inosine to bind with all four nucleotides was not crucial for the observed increase in microbial richness. Instead, differences in community compositions were correlated with the identity of the nearest-neighbor 3' of the primer-targeting region. By influencing the thermal stability of primer hybridization, this position may play a previously unrecognized role in biased amplification of 16S rRNA gene sequences. In conclusion, the combined use of inosine and unmodified primers enables the complementary retrieval of 16S rRNA gene types, thereby expanding the observed diversity of complex microbial communities. PMID- 19807789 TI - Controlled mixed culture fermentation: a new perspective on the use of non Saccharomyces yeasts in winemaking. AB - Mixed fermentations using controlled inoculation of Saccharomyces cerevisiae starter cultures and non-Saccharomyces yeasts represent a feasible way towards improving the complexity and enhancing the particular and specific characteristics of wines. The profusion of selected starter cultures has allowed the more widespread use of inoculated fermentations, with consequent improvements to the control of the fermentation process, and the use of new biotechnological processes in winemaking. Over the last few years, as a consequence of the re evaluation of the role of non-Saccharomyces yeasts in winemaking, there have been several studies that have evaluated the use of controlled mixed fermentations using Saccharomyces and different non-Saccharomyces yeast species from the wine environment. The combined use of different species often results in unpredictable compounds and/or different levels of fermentation products being produced, which can affect both the chemical and the aromatic composition of wines. Moreover, possible synergistic interactions between different yeasts might provide a tool for the implementation of new fermentation technologies. Thus, knowledge of the Saccharomyces and non-Saccharomyces wine yeast interactions during wine fermentation needs to be improved. To reach this goal, further investigations into the genetic and physiological background of such non-Saccharomyces wine yeasts are needed, so as to apply '-omics' approaches to mixed culture fermentations. PMID- 19807790 TI - Human peripheral gammadelta T cells possess regulatory potential. AB - Deficiency in gammadelta T cells aggravates colitis in animal models suggesting that gammadelta T cells have regulatory properties. Therefore, proliferation, suppression and cytokine secretion of human gammadelta T cells were determined in vitro. Human peripheral gammadelta T cells were isolated from the whole blood of healthy donors by magnetic antibody cell sorting technology. The proliferation after CD3/CD28 stimulation was measured by (3)[H]thymidine incorporation. Interferon-gamma (IFN-gamma), interleukin-2 (IL-2), transforming growth factor beta (TGF-beta) and IL-10 concentrations were measured by enzyme-linked immunosorbent assay; TGF-beta messenger RNA was also measured by reverse transcription-polymerase chain reaction. The expression of latency associated peptide (LAP), a TGF-beta complex component, intracellular cytokine content and T helper cell proliferation were measured by flow cytometry. Human gammadelta T cells showed poor proliferation upon CD3/CD28 stimulation and suppressed T helper cell growth stronger than CD4(+) CD25(+) T cells, although gammadelta T cells were FOXP3 negative. They secreted little IL-2 but high concentrations of IFN gamma, IL-10 and TGF-beta. When looking at LAP expression the Vdelta1 subset was found to be the main TGF-beta producer compared to Vdelta2 T cells. Taken together, peripheral gammadelta T cells have in vitro a more potent regulatory potential than CD4(+) CD25(+) cells regarding T helper cell suppression. This is most likely the result of strong TGF-beta secretion, particularly by the Vdelta1 subset. PMID- 19807791 TI - Mediterranean diet and high dietary acid load associated with mixed nuts: effect on bone metabolism in elderly subjects. AB - OBJECTIVES: To analyze the effect of differing diet on the acid load content on bone metabolism. DESIGN: Multicentric, randomized, single-blind, parallel-group clinical trial. SETTING: Outpatient clinics. PARTICIPANTS: Two hundred thirty eight elderly men and women aged 60 to 80 at high risk for cardiovascular disease were randomly assigned to three interventional groups: a recommended low-fat diet (control diet group), a Mediterranean diet supplemented with virgin olive oil, or a Mediterranean diet supplemented with mixed nuts. MEASUREMENTS: Main outcomes were 12-month changes from baseline in bone formation and resorption markers and bone mass measured according to quantitative ultrasound scanning. RESULTS: The baseline data on the anthropometric, bone densitometry, and biochemical variables did not differ between the three groups. Dietary potential renal acid load (PRAL) and daily net endogenous acid production (NEAP) at baseline did not differ between groups. After intervention, subjects allocated to the Mediterranean diet with mixed nuts had a significant increase of PRAL and NEAP. In comparison, subjects in the Mediterranean diet with nuts group had higher parathyroid hormone (PTH) levels (2.63, 95% confidence interval (CI)=-1.01-6.35, P=.02) and a nonsignificantly higher (0.31, 95% CI=-0.13-0.74, P=.14) urine free deoxypyridoxine:creatinine ratio, a marker of bone resorption, than the control group and the Mediterranean diet with virgin olive oil group. CONCLUSION: A Mediterranean dietary pattern associated with a high dietary acid load derived from consumption of mixed nuts does not seem to have a much greater effect on bone metabolism biomarkers, with the exception of PTH levels, than a Mediterranean diet without mixed nuts or a control diet in elderly subjects. PMID- 19807792 TI - Multisite geriatrics clerkship for fourth-year medical students: a successful model for teaching the Association of American Medical Colleges' core competencies. AB - As the population ages, it is important that graduating medical students be properly prepared to treat older adults, regardless of their chosen specialty. To this end, the Association of American Medical Colleges (AAMC) and the John A. Hartford Foundation convened a consensus conference to establish core competencies in geriatrics for all graduating medical students. An ambulatory geriatric clerkship for fourth-year medical students that successfully teaches 24 of the 26 AAMC core competencies using an interdisciplinary, team-based approach is reported here. Graduating students (N=158) reported that the clerkship was successful at teaching the core competencies, as evidenced by positive responses on the AAMC Graduation Questionnaire (GQ). More than three-quarters (80-93%) of students agreed or strongly agreed that they learned the seven geriatrics concepts asked about on the GQ, which cover 14 of the 26 core competencies. This successful model for a geriatrics clerkship can be used in many institutions to teach the core competencies and in any constellation of geriatric ambulatory care sites that are already available to the faculty. PMID- 19807793 TI - Case report: combined use of donepezil and galantamine in mixed dementia. PMID- 19807794 TI - Event-free 6 months of an octogenarian with an undeployed stent in the left main coronary trunk: efficacy of dual antiplatelet therapy. PMID- 19807795 TI - Chronic kidney disease is a potential risk factor for the development of purple urine bag syndrome. PMID- 19807796 TI - Chair rise ability and length of stay in hospitalized older adults. PMID- 19807797 TI - A rare coronary artery anomaly in a patient with isolated congenitally corrected transposition of great arteries. PMID- 19807798 TI - Premature condemnation of clinical decision support as a useful tool for patient safety in computerized provider order entry. PMID- 19807799 TI - Statins reduce cardiovascular events in primary and secondary prevention trials without causing an increase in carcinoma. PMID- 19807801 TI - Whole-diet approach: working on a criterion validity for age-related cognitive decline and mild cognitive impairment. PMID- 19807802 TI - Clinical and therapeutic aspects in elderly patients with Merkel cell carcinoma: special focus on radiotherapy. PMID- 19807804 TI - If shrimps had lips, they could whistle ... PMID- 19807805 TI - Early relapse of transient left ventricular apical ballooning syndrome in an 84 year-old woman. PMID- 19807806 TI - Primary care providers' assessment of older Latino patients' instrumental activities of daily living ability: implications for improving quality of care. PMID- 19807807 TI - Type IV dual left anterior descending artery in a septagenarian. PMID- 19807808 TI - A short nocturnal activity scale used by nurses to evaluate hospitalized patients with dementia. PMID- 19807809 TI - Survival and mortality in older adults living at high altitude in Bolivia: a preliminary report. PMID- 19807810 TI - Delirium in nursing home residents across care transitions: a preliminary report. PMID- 19807811 TI - Patients with solitary infarcts may be more responsive to vascular care. PMID- 19807812 TI - Improvement of swallowing reflex after electrical stimulation to lower leg acupoints in patients after stroke. PMID- 19807813 TI - What academic internal medicine subspecialists identify as important geriatric learning needs for residents: a pilot study. PMID- 19807814 TI - Zolpidem dependence in a geriatric patient: a case report. PMID- 19807815 TI - Walker use affects Timed Up and Go and gait speed measures. PMID- 19807816 TI - Assessment of gastric motility using magnetic resonance imaging in patients receiving enteral nutrition. PMID- 19807820 TI - Patterns of bone loss around teeth restored with endodontic posts. AB - OBJECTIVES: This retrospective study described the pattern of bone loss around teeth with endodontic posts in periodontitis patients, and compared it with contra-lateral teeth without posts. MATERIAL AND METHODS: From full-mouth radiographic surveys of 146 periodontitis patients (> or =35 years), 194 roots with endodontic posts and contra-laterals without posts were selected. Upper molars, pre-molars with two posts and roots of lower molars with two posts were excluded. Technical parameters of the post space preparation, endodontic and restorative status were evaluated. The level of alveolar bone measured in millimetre from the cemento-enamel junction (CEJ)/restoration margin and the pattern of bone loss (angular/horizontal) were evaluated on both mesial and distal aspects of roots with posts and contra-laterals, but not on the furcal areas of lower molars. RESULTS: The distance from the bone level to the CEJ/restoration margin was similar for teeth with posts and contra-laterals. However, teeth with posts had more angular defects mesially (18.8%versus 7.3%) as compared with their contra-laterals without posts. The defects around teeth with posts appeared to be typical in the sense that their apical level approximated the tip of the endodontic post. CONCLUSION: In periodontitis patients, teeth restored by an endodontic post had angular bony defects on the mesial aspect more frequently in comparison with their contra-laterals. PMID- 19807821 TI - Open flap debridement and guided tissue regeneration after 10 years in infrabony defects. AB - OBJECTIVE: Evaluation of the 10-year results after open flap debridement (OFD) and guided tissue regeneration (GTR) therapy of infrabony defects in a randomized controlled clinical trial. MATERIALS AND METHODS: In 16 periodontitis patients OFD or polylactide acetyltributyl citrate barriers (GTR; n=23) were assigned randomly to 44 infrabony defects. In a subgroup of 10 patients exhibiting 2 contra-lateral defects each OFD and GTR was assigned to either side (split mouth). At baseline, 12, and 120 +/- 12 months after surgery clinical parameters were obtained. RESULTS: Fifteen patients (41 defects) were available at 120 months. Twelve and 120 +/- 12 months after therapy both groups showed statistically significant (p<0.01) attachment gain (split-mouth: OFD: 12 months: 3.60 +/- 2.67 mm; 120 months: 3.65 +/- 3.36 mm; GTR: 12 months: 3.50 +/- 1.90 mm; 120 months: 2.85 +/- 2.24 mm; parallel: OFD: 12 months: 3.47 +/- 2.80 mm; 120 months: 3.41 +/- 2.75 mm; GTR: 12 months: 3.67 +/- 2.11 mm; 120 months: 2.89 +/- 2.12 mm). From 12 to 120 months both groups experienced insignificant attachment changes, however, six teeth (two OFD, four GTR) were lost (all for prosthodontic reasons). The study failed to show statistically significant attachment gain differences between both groups after 120 months. CONCLUSIONS: Ten years after OFD and GTR in infrabony defects 35 of 41 teeth were still in place. PMID- 19807822 TI - Histopathology and immunohistochemistry of depigmented lesions in lupus erythematosus. AB - BACKGROUND: Depigmented lesions (DL) have been described in areas previously damaged by inflammation in lupus erythematosus (LE). In the absence of typical lesions, distinction with other achromic diseases might be challenging. We studied the histological features and the behavior of melanocytes in these lesions. METHODS: Tissue sections of 12 patients with lupus and DL were stained with hematoxylin-eosin, periodic acid-Schiff and Fontana-Masson. Melanocytes were counted by immunohistochemistry methods using Melan A and HMB-45. Ten biopsies of normal skin were used as controls. RESULTS: The most common histological findings were: cellular infiltration (75%); hyperkeratosis (66.7%); thickening of basement membrane (66.7%); thinning and flattening of the epidermis (58.3%) and degenerative changes in collagen fibers (50%). Epidermal melanin and melanocytes were found in 41.7%. The melanocyte counts by HMB-45 and Melan A were significantly lower than in normal skin. CONCLUSIONS: The DL still fulfill histological criteria for lupus. In the absence of a precise histological diagnosis, thickening of basement membrane, hyperkeratosis, cellular infiltration, epidermal atrophy and elastosis are the most common features. Loss of melanocytes and the dermal fibrosis suggests that DL in cutaneous LE behave as post-inflammatory scars. PMID- 19807823 TI - Morphological and immunohistochemical characteristics of atypical fibroxanthoma with a special emphasis on potential diagnostic pitfalls: a review. AB - The present manuscript gives emphasis on recognizing different morphological variants of atypical fibroxanthoma (AFX), on validation of immunohistochemical markers and on discussing potential diagnostic pitfalls. MATERIAL AND METHODS: Histological features analyzed in 66 AFXs were: ulceration, morphological variants, growth pattern, location in the skin and vascular/perineural invasion. The antibodies used were CK-MNF116, CK-AE1/AE3, S100, smooth muscle actin, desmin, CD31 and EMA. RESULTS: The study included 59 males, 7 females, aged 55-95 years, mean 77 years. All developed on sun damaged skin. Ulceration was present in 50%. Morphological patterns were pleomorphic spindle and epithelioid cells (60.6%), predominantly spindle cells (19.7%), purely spindle-cells (13.6%), and predominantly epithelioid cells (6.1%). Most were localized in the dermis (57.6%). An expansile (36.4%) rather than infiltrative (6.1%) growth into superficial subcutis was also noted. No vascular/perineural invasion was seen. Additional changes were hemorrhagic and pseudoangiomatous areas (24.2%), granular cell change (22.7%), keloid-like areas (9.1%), myxoid change (7.6%), osteoclast like giant cells (6.1%) and clear cell change (4.6%). AFXs were consistently negative for S100, CK-MNF116, CK-AE1/AE3 and desmin. Focal positivity for SMA (45.2%), EMA (24.4%) and CD 31 (9.5%) was seen. CONCLUSIONS: A diagnosis of AFX is still made by exclusion of other malignant neoplasms with similar morphology. Immunohistochemistry plays a crucial role in this distinction, but can also be misleading. This study expands the spectrum of non-vascular CD31 positive tumors. PMID- 19807824 TI - Erythema gyratum repens, not always a bad omen for patients. PMID- 19807825 TI - Treatment of nail psoriasis with adalimumab: an open label unblinded study. AB - BACKGROUND: Despite numerous advances in the therapeutic management of cutaneous psoriasis, there is a lack of standardized therapeutic regimens for psoriatic nail disease. OBJECTIVE: An open, non-randomized, unblinded study was designed to evaluate the efficacy and safety of adalimumab in the treatment of nail psoriasis. PATIENTS/METHODS: Seven patients suffering from severe plaque-type psoriasis and 14 with psoriatic arthritis and cutaneous psoriasis with concomitant nail involvement were enrolled into the study. The applied dose regimen of adalimumab was the same as the one recommended for cutaneous psoriasis. Outcome measures were assessed at baseline and at weeks 12 and 24 using the Nail Psoriasis Severity Index (NAPSI). Patients also filled in a Greek translation of the international onychomycosis-specific questionnaire to assess the impact of the nail improvement on their quality of life. RESULTS: All 21 patients completed the study and were eligible for statistical analysis. Significant improvement was recorded after the eighth injection. Mean NAPSI (NAPSIm) at baseline was 10.57 +/- 1.21 for the fingernails and 14.57 +/- 2.50 for the toenails in patients with just cutaneous psoriasis and 23.86 +/- 2.00 for the fingernails and 29.29 +/- 2.87 for the toenails in patients with psoriatic arthritis. NAPSIm at week 12 was 5.57 +/- 0.78 for the fingernails and 9.57 +/- 2.17 for the toenails in patients with just cutaneous psoriasis and 12.86 +/- 1.05 for the fingernails and 19.21 +/- 2.07 for the toenails in patients with psoriatic arthritis. NAPSIm after 24 weeks of treatment was 1.57 +/- 0.20 for the fingernails and 4.14 +/- 1.58 for the toenails in patients with cutaneous psoriasis and 3.23 +/- 0.32 for the figernails and 10.00 +/- 1.40 for the toenails in patients with psoriatic arthritis. Treatment was well tolerated with minimal and temporary side-effects limited to the site of injection. All patients were satisfied, while marked improvement in their quality of life was recorded based on the reduction of the scores obtained from the international quality of life questionnaire. CONCLUSIONS: Despite the lack of a control group, our results demonstrate a beneficial effect of adalimumab on psoriatic nail disease. PMID- 19807826 TI - Quality of life and psychological adaptation of Korean adolescents with vitiligo. AB - AIM: The aim of the study was to evaluate the impact of vitiligo on the quality of life and psychological adaptation in a Korean adolescent population. METHODS: Fifty-seven adolescents aged 12 to 18 years with vitiligo were evaluated using self-report scales, namely the Skindex-29, Piers-Harris self-concept, Center for Epidemiologic Studies Depression Scale (CES-D), and Revised Children's Manifest Anxiety Scale (RCMAS). RESULTS: Mean Skindex-29 subscales were as follow; 21.8 (global), 16.3 (symptom), 18.6 (function) and 29.5 (emotion). Several clinical variables, such as duration of vitiligo, facial involvement, history of previous treatment, and patient-assessed severity, affected the Skindex-29 subscales in various ways. However, differences in Skindex-29 scores according to the type of vitiligo, extent of involvement, and family history were not observed. The Piers Harris self-concept scores showed a negative correlation with Skindex-29 scores, while other psychological measures (CES-D and RCMAS) were positively correlated. CONCLUSION: The quality of life of adolescents with vitiligo is closely related to the patients' apprehensions about their disease, psychosocial adjustment, and psychiatric morbidity, rather than the clinical severity of the condition itself. Clinicians should recognize and deal with psychological adaptation along with medical intervention when treating adolescent patients with vitiligo. PMID- 19807827 TI - Photoepilation with variable pulsed light in non-facial body areas: evaluation of efficacy and safety. AB - BACKGROUND: Variable Pulsed Light (VPL) is a new intensity pulse light (IPL) system. We evaluated VPL for safety and long-term efficacy in non-facial hair removal. STUDY DESIGN/MATERIAL AND METHODS: The Variable Pulsed Light [VPL System (Energist Limited, UK), 610-950 nm] system was used for the removal of unwanted non-facial dark pigmented body hair in 42 volunteers (40 females, two males, age: 18-43 years, mean: 30.4 +/- 7.4 years) with Fitzpatrick skin phototype (SPT) II (33.3%), III (38.1%) and IV (28.6%), and brown (43.8%) or black (56.2%) fine (19.0%), medium (47.6%) or coarse (9.4%) body hair. Minimum follow-up period was 12 months (mean: 13.14 +/- 2.38). Treatment parameters were as follows: number x duration of micropulses: 15 x 5-15 x 7 ms, pulse delay: 1-8 ms, fluence: 30-45 J/cm(2). RESULTS: Very good (> or =76% hair reduction) and good treatment efficacies (hair reduction: 51-75%) were observed in the majority of treated patients (n = 18; 42.9%; n = 14; 33.3% respectively), four patients (9.5%) had only moderate (hair reduction: 26-50%) and six (14.3%) patients had no (hair reduction < or =25%) treatment response. Side-effects (reversible erythema, leukotrichia) were sparse. More favourable long-term therapy outcomes were observed for higher SPT (III and IV), medium or coarse black hair and younger patients. CONCLUSION: The VPL System is a relatively efficient and safe treatment for long-term hair removal. Higher SPT (III and IV) with black and medium to coarse hair, and younger age of the patients appear to be factors of significant importance predicting long-term efficacy. PMID- 19807828 TI - Guidelines for practical use of MAL-PDT in non-melanoma skin cancer. AB - Methyl aminolaevulinate photodynamic therapy is increasingly practiced in the treatment of actinic keratoses, Bowen's disease and basal cell carcinomas. This method is particularly suitable for treating multiple lesions, field cancerization and lesions in areas where a good cosmetic outcome is of importance. Good treatment routines will contribute to a favourable result. The Norwegian photodynamic therapy (PDT) group consists of medical specialists with long and extensive PDT experience. With support in the literature, this group presents guidelines for the practical use of topical PDT in non-melanoma skin cancer. PMID- 19807829 TI - Natural and sexual selection against immigrants maintains differentiation among micro-allopatric populations. AB - Local adaptation to divergent environmental conditions can promote population genetic differentiation even in the absence of geographic barriers and hence lead to speciation. But what mechanisms contribute to reproductive isolation among diverging populations? We tested for natural and sexual selection against immigrants in a fish species inhabiting (and adapting to) nonsulphidic surface habitats, sulphidic surface habitats and a sulphidic cave. Gene flow is strong among sample sites situated within the same habitat type, but low among divergent habitat types. Our results indicate that females of both sulphidic populations discriminate against immigrant males during mate choice. Furthermore, using reciprocal translocation experiments, we document natural selection against migrants between nonsulphidic and sulphidic habitats, whereas migrants between sulphidic cave and surface habitats did not exhibit increased mortality within the same time period. Consequently, both natural and sexual selection may contribute to isolation among parapatric populations, and selection against immigrants may be a powerful mechanism facilitating speciation among locally adapted populations even over very small spatial distances. PMID- 19807830 TI - Reproductive barriers between two sympatric beetle species specialized on different host plants. AB - Knowledge on interspecific pre- and post-zygotic isolation mechanisms provides insights into speciation patterns. Using crosses (F(1) and backcrosses) of two closely related flea beetles species, Altica fragariae and A. viridicyanea, specialized on different hosts in sympatry, we measured: (a) the type of reproductive isolation and (b) the inheritance mode of preference and host specific performance, using a joint-scaling test. Each species preferred almost exclusively its host plant, creating strong prezygotic isolation between them, and suggesting that speciation may occur at least partly in sympatry. Reproductive isolation was intrinsic between females of A. fragariae and either A. viridicyanea or F(1) males, whereas the other crosses showed ecologically dependent reproductive isolation, suggesting ecological speciation. The genetic basis of preference and performance was at least partially independent, and several loci coded for preference, which limits the possibility of sympatric speciation. Hence, both ecological and intrinsic factors may contribute to speciation between these species. PMID- 19807831 TI - Abstracts of the Australia & New Zealand Medical & Surgical Gastrointestinal Week 2009. October 21-24, 2009. Sydney, NSW, Australia. PMID- 19807844 TI - Toward an implantable wireless cardiac monitoring platform integrated with an FDA approved cardiovascular stent. AB - Continuous monitoring of blood pressure from a minimally invasive device in the pulmonary artery can serve as a diagnostic and early warning system for cardiac health. The foremost challenge in such a device is the wireless transfer of data and power from within the blood vessel to an external device while maintaining unrestricted flow through the artery. We present a miniaturized system, which is attached to the outer surface of a regular or drug-eluting FDA-approved stent. When expanded, the stent maintains a patent vessel lumen while allowing contact between the electronic sensors and the blood supply. The stent-based antenna can be used for both wireless telemetry and power transfer for the implanted electronics. Using the stent platform as both a radiating antenna and a structural support allows us to take advantage of an FDA-approved device whose safety and surgical procedure are well established. The electronics package has been reduced to an area of less than 1 mm(2), with a thickness under 300 mum. A minimally invasive implantation procedure allows the delivery of the stent-based implant in nearly any major vessel of the body. This article describes an initial prototype with two stents configured as a single dipole, a 2.4-GHz transmitter microchip, and a battery, and validates transcutaneous transmission through ex vivo and in vivo porcine studies. The results demonstrate the feasibility of a stent-based wireless platform for continuous monitoring of blood pressure. PMID- 19807845 TI - Nitric oxide and cyclic nucleotide signal transduction modulates synaptic vesicle turnover in human model neurons. AB - The human Ntera2 (NT2) teratocarcinoma cell line can be induced to differentiate into post-mitotic neurons. Here, we report that the human NT2 neurons generated by a spherical aggregate cell culture method express increasing levels of typical pre-synaptic proteins (synapsin and synaptotagmin I) along the neurite depending on the length of in vitro culture. By employing an antibody directed against the luminal domain of synaptotagmin I and the fluorescent dye N-(3 triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide, we show that depolarized NT2 neurons display calcium-dependent exo-endocytotic synaptic vesicle recycling. NT2 neurons express the neuronal isoform of neuronal nitric oxide synthase and soluble guanylyl cyclase (sGC), the major receptor for nitric oxide (NO). We tested whether NO signal transduction modulates synaptic vesicle turnover in human NT2 neurons. NO donors and cylic guanosine-monophosphate analogs enhanced synaptic vesicle recycling while a sGC inhibitor blocked the effect of NO donors. Two NO donors, sodium nitroprusside, and and N-Ethyl-2-(1 ethyl-2-hydroxy-2-nitrosohydrazino) ethanamine evoked vesicle exocytosis which was partially blocked by the sGC inhibitor. The activator of adenylyl cyclase, forskolin, and a cAMP analog induced synaptic vesicle recycling and exocytosis via a parallel acting protein kinase A pathway. Our data from NT2 neurons suggest that NO/cyclic nucleotide signaling pathways may facilitate neurotransmitter release in human brain cells. PMID- 19807846 TI - An interaction of oxytocin receptors with metabotropic glutamate receptors in hypothalamic astrocytes. AB - Hypothalamic astrocytes play a critical role in the regulation and support of many different neuroendocrine events, and are affected by oestradiol. Both nuclear and membrane oestrogen receptors (ERs) are expressed in astrocytes. Upon oestradiol activation, membrane-associated ER signals through the type 1a metabotropic glutamate receptor (mGluR1a) to induce an increase of free cytoplasmic calcium concentration ([Ca(2+)](i)). Because the expression of oxytocin receptors (OTRs) is modulated by oestradiol, we tested whether oestradiol also influences oxytocin signalling. Oxytocin at 1, 10, and 100 nm induced a [Ca(2+)](i) flux measured as a change in relative fluorescence [DeltaF Ca(2+) = 330 +/- 17 relative fluorescent units (RFU), DeltaF Ca(2+) = 331 +/- 22 RFU, and DeltaF Ca(2+) = 347 +/- 13 RFU, respectively] in primary cultures of female post-pubertal hypothalamic astrocytes. Interestingly, OTRs interacted with mGluRs. The mGluR1a antagonist, LY 367385 (20 nm), blocked the oxytocin (1 nm) induced [Ca(2+)](i) flux (DeltaF Ca(2+) = 344 +/- 19 versus 127 +/- 11 RFU, P < 0.001). Conversely, the mGluR1a receptor agonist, (RS)-3,5-dihydroxyphenyl glycine (100 nm), increased the oxytocin (1 nm)-induced [Ca(2+)](i) response (DeltaF Ca(2+) = 670 +/- 31 RFU) compared to either compound alone (P < 0.001). Because both oxytocin and oestradiol rapidly signal through the mGluR1a, we treated hypothalamic astrocytes sequentially with oxytocin and oestradiol to determine whether stimulation with one hormone affected the subsequent [Ca(2+)](i) response to the second hormone. Oestradiol treatment did not change the subsequent [Ca(2+)](i) flux to oxytocin (P > 0.05) and previous oxytocin exposure did not affect the [Ca(2+)](i) response to oestradiol (P > 0.05). Furthermore, simultaneous oestradiol and oxytocin stimulation failed to yield a synergistic [Ca(2+)](i) response. These results suggest that the OTR signals through the mGluR1a to release Ca(2+) from intracellular stores and rapid, nongenomic oestradiol stimulation does not influence OTR signalling in astrocytes. PMID- 19807847 TI - Palatable high-energy diet decreases the expression of cannabinoid type 1 receptor messenger RNA in specific brain regions in the rat. AB - In laboratory rodents, a palatable high-energy diet (PHED) is usually consumed in a higher quantity than a standard laboratory diet, leading to the development of an obese phenotype. The central effects of PHED are not fully understood. Nonetheless, the long-term consumption of PHED can decrease cannabinoid type 1 receptor (CB1R) protein density in particular brain regions. However, little is known about the diet-dependent regulation of the brain expression of CB1R mRNA. The present study aimed to investigate the effects of the long-term consumption of PHED and short-term (12 h) food deprivation on the brain expression of CB1R mRNA. For 13 weeks, rats were fed a standard laboratory chow or PHED presented as a free choice of chow, shortcake biscuits and pork spread. In total, the food intake of PHED rats was higher than that of chow-fed animals. Expectedly, PHED rats demonstrated higher body weight than chow-fed animals. The difference in body weight between PHED- and chow-fed rats was as result of the fat but not the lean mass. PHED-fed rats had significantly higher plasma levels of leptin and insulin and significantly higher levels of expression of suppressor of cytokine signalling 3 (SOCS-3) in the arcuate hypothalamic nucleus. The long-term consumption of PHED significantly decreased the levels of CB1R mRNA expression in the cingulate (Cg) cortex, ventromedial hypothalamic nucleus and the descending/autonomic divisions of the parvocellular hypothalamic nucleus (PVH), the ventrolateral parvocellular PVH and, to a lesser extent, the dorsomedial parvocellular PVH. Acute food deprivation decreased the levels of CB1R transcript in the Cg and ventrolateral parvocellular PVH. Altogether, the present results demonstrate that long-term PHED leads to an increase in the hypothalamic expression of SOCS-3 mRNA and a decrease in expression of CB1R mRNA in the Cg cortex and specific hypothalamic regions. PMID- 19807849 TI - Glucokinase and glucokinase regulatory proteins are functionally coexpressed before birth in the rat brain. AB - Our previous description of functional glucokinase (GK) isoforms and their interactions with glucokinase regulatory protein (GKRP) in adult rat and human brains suggested that both participate in glucose sensing in the central nervous system. To determine whether both proteins are coexpressed and active before birth or during early post-natal life, we characterised these molecules in the brains of foetal and post-natal pup rats. We found GK and GKRP mRNAs that were similar to those previously reported in the adult rat brain. Likewise, GK and GKRP gene expression gave rise to proteins of 52 and 69 kDa, respectively. Immunohistochemistry experiments showed the colocalisation of both GK and GKRP proteins in the same brain cells of 21-day-old rat foetuses. Furthermore, coprecipitation of GK and GKRP in the presence of fructose 6-phosphate suggests interactions between both proteins. The presence of GK phosphorylating activity was detected in different brain areas of 21-day-old foetuses with a contribution to the total glucose-phosphorylating activity of between 17.2 +/- 1.7% and 12.4 +/- 3.7%, with the hypothalamus being the region of maximum activity. The hypothalamic GK activity in 21-day-old foetuses has a high apparent K(m) for glucose and no product inhibition by glucose 6-phosphate. Our findings indicate that both proteins may be functionally active before birth and that they can act within a glucose sensor system involved in controlling food intake. PMID- 19807848 TI - Gene expression profiles of intracellular and membrane progesterone receptor isoforms in the mediobasal hypothalamus during pro-oestrus. AB - Progesterone action is mediated by its binding to specific receptors. Two progesterone receptor (PR) isoforms (PRA and PRB), three membrane progesterone receptor (mPR) subtypes (mPRalpha, mPRbeta and mPRgamma) and at least one progesterone membrane-binding protein [PR membrane component 1 (PRmc1)] have been identified in reproductive tissues and brain of various species. In the present study, we examined gene expression patterns for PR isoforms, mPR subtypes and PRmc1 in the rat mediobasal hypothalamus (MBH) during pro-oestrus. The mRNA level for each receptor subtype was quantified by a real-time reverse transcriptase polymerase chain reaction (RT-PCR) at the time points: 13.00 h on dioestrous day 2; 09.00, 13.00, 17.00 and 22.00 h on pro-oestrus; and 13.00 h on oestrus. For PR, one primer set amplified PRA+PRB, whereas a second primer set amplified PRB. As expected, PRA+PRB mRNA expression was greater than PRB in MBH tissue. PRB mRNA levels increased throughout the day on pro-oestrus, with the highest levels being observed at 17.00 h. PRB mRNA levels in the MBH were increased by 2.4- and 3.0 fold at 13.00 and 17.00 h, respectively, on pro-oestrus compared to 13.00 h on dioestrous day 2. There were differential mRNA expression levels for mPRs and PRmc1 in the MBH, with the highest expression for PRmc1 and the lowest for mPRgamma. The mPRalpha mRNA contents at 13.00 and 17.00 h on pro-oestrus were increased by 1.5-fold compared to that at 13.00 h on dioestrous day 2. The mPRbeta mRNA levels at 13.00 and 17.00 h on pro-oestrus were 2.5- and 2.4-fold higher compared to that at 13.00 h on dioestrous day 2, respectively. PRA+PRB, mPRgamma and PRmc1 mRNA levels did not vary on pro-oestrus. These findings suggest that the higher expression of PRB, mPRalpha and mPRbeta in the MBH on pro oestrous afternoon may influence both genomic and nongenomic mechanisms of progesterone action during the critical pre-ovulatory period. PMID- 19807850 TI - Consensus conference on intracranial atherosclerotic disease: rationale, methodology, and results. AB - The consensus conference on intracranial atherosclerotic disease (ICAD) identifies principles of management, and research priorities in various aspects upon which leading experts can agree (using "Delphi" method). ICAD is more prevalent in Asian, Hispanic, and African-American populations. Patients who have had a stroke or transient ischemic attack (TIA) attributed to stenosis (50-99%) of a major intracranial artery face a 12-14% risk of subsequent stroke during the 2-year period after the initial ischemic event, despite treatment with antithrombotic medications. The annual risk of subsequent stroke may exceed 20% in high-risk groups. The medical treatment of patients with symptomatic ICAD is directed toward: 1. Prevention of intraluminal thrombo-embolism, 2. plaque stabilization and regression, and 3. management of atherogenic risk factors. In patients with ICAD, short-term and long-term anticoagulation (compared with aspirin) have not shown to be beneficial. The current guidelines recommend that aspirin monotherapy, the combination of aspirin and extended release dipyridamole, and clopidogrel monotherapy (rather than oral anticoagulants) are all acceptable options in patients with non-cardioembolic ischemic stroke and TIA. Overall, the subgroup analysis from randomized trials provides evidence about benefit of aggressive atherogenic risk factor management among patients with ICAD. Intracranial angioplasty with or without stent placement has evolved as a therapeutic option for patients with symptomatic ICAD, particularly those with high-grade stenosis with recurrent ischemic symptoms and/or medication failure. A matched comparison between medical-treated patients in the Warfarin Aspirin Symptomatic Intracranial Disease (WASID) study and stent-treated patients in the National Institutes of Health intracranial stent registry concluded that stent placement may offer benefit in patients with 70-99% stenosis. The 5-year, multicenter, prospective, randomized Stenting and Aggressive Medical Management for Preventing Recurrent stroke in Intracranial Stenosis study supported by the National Institutes of Health is currently comparing stent placement with intense medical management with intense medical management alone in patients with high grade symptomatic intracranial stenosis. The proceedings of the consensus conference provide a template for standardizing management of patients with ICAD and determining research priorities. PMID- 19807851 TI - Epidemiology of intracranial stenosis. AB - Intracranial stenosis is a common etiology for ischemic stroke. Due to limitations of imaging studies, there are limited data on the prevalence of symptomatic and asymptomatic intracranial stenosis. Intracranial stenosis is more prevalent in Asian, Hispanic, and African-American populations. The reported proportion of patients with symptomatic intracranial stenosis among those hospitalized for ischemic cerebral events varies from 1% in non-Hispanic whites to as high as 50% in Asian populations. In population-based studies, the estimated prevalence of symptomatic intracranial disease varies from 1 in 100,000 for whites to 15 in 100,000 in African Americans. A Chinese population-based study reported intracranial stenosis in 7% of the population aged more than 40 years. Autopsy studies have noted intracranial atherosclerotic disease in about 23% of population in the 6th decade and 80% of population in the 9th decade of life. Angiotensin-converting enzyme polymorphisms, plasma endostatin/vascular endothelial growth factor ratio, glutathione S-transferase omega-1 gene polymorphism, and plasma homocysteine levels are non-modifiable risk factors noted to be associated with intracranial stenosis. Hypertension and serum lipid profile are major modifiable risk factors, whereas sickle cell disease is an uncommon risk factor that can be managed to reduce risk. Associations of intracranial atherosclerosis with diabetes mellitus, metabolic syndrome, Alzheimer's disease, aortic plaques, radiotherapy, and meningitis are less well documented. PMID- 19807852 TI - Natural history of asymptomatic intracranial arterial stenosis. AB - The prevalence and natural history of asymptomatic intracranial arterial stenosis are not very well characterized. Existing data suggest that incidentally discovered asymptomatic intracranial stenosis presents a fairly low risk of stroke, though substantial uncertainty remains. Patients may be at greater risk if there are tandem stenoses. Methods to stratify the risk of stroke with asymptomatic intracranial atherosclerotic lesions have yet to be established and validated. In general, aggressive intervention for an asymptomatic intracranial stenosis is not currently recommended. PMID- 19807853 TI - Natural history of symptomatic intracranial arterial stenosis. AB - Patients who have had a stroke or transitory ischemic attack (TIA) attributed to stenosis (50-99%) of a major intracranial artery face a 12-14% risk of subsequent stroke during the 2-year period after the initial ischemic event, despite treatment with antithrombotic medications. Most of this risk accrues during the first year. Some patients are at substantially greater risk, particularly those with a severe (70-99%) stenosis, those who have recently had an ischemic event, and women. Patients may also be at high risk if they had an initial stroke rather than TIA or if they have symptoms precipitated by hemodynamic maneuvers. The annual risk of subsequent stroke may exceed 20% in these high-risk groups. PMID- 19807854 TI - Diagnosis and quantitation of intracranial stenosis. AB - Noninvasive tests, transcranial Doppler, magnetic resonance angiography and computed tomography angiography, and invasive catheter angiography are available to diagnose the presence and quantify the severity of intracranial atherosclerotic stenosis. Each test has its advantages and disadvantages, including the gold standard of catheter angiography. Careful use and measurements with any test are crucial to optimal research and treatment of patients suspected of harboring intracranial atherosclerotic stenosis. Rigorous trials assessing the performance of these tests are few in number. The design and results of the SONIA trial are reviewed. The clinical utility of the SONIA approach and data set, in both clinical and research settings, is reviewed. PMID- 19807855 TI - Medical treatment of patients with intracranial atherosclerotic disease. AB - The medical treatment of patients with symptomatic intracranial atherosclerotic disease (ICAD) is directed toward reducing the risk of new ischemic events. The overall strategy is divided into: (1) prevention of occurrence of intraluminal thrombus, with or without embolism; (2) plaque stabilization and regression; and (3) management of atherogenic risk factors. In patients with ICAD, short-term and long-term anticoagulation (compared with aspirin) has not shown to be beneficial. The current guidelines recommend that aspirin monotherapy, the combination of aspirin and extended release dipyridamole, and clopidogrel monotherapy (rather than oral anticoagulants) are all acceptable options in patients with noncardioembolic ischemic stroke and transient ischemic attack. The findings of another pilot trial suggest that symptomatic ICAD is a dynamic lesion and cilostazol may prevent its progression. Overall, the subgroup analysis from randomized trials, provide evidence about benefit of aggressive atherogenic risk factor management among patients with ICAD. Current guidelines recommend statin therapy with intensive lipid-lowering effects for patients with atherosclerotic ischemic stroke or transient ischemic attack with or without known coronary artery disease to reduce the risk of stroke and cardiovascular events. PMID- 19807856 TI - The evolution and role of endovascular therapy for the treatment of intracranial atherosclerotic disease. AB - Intracranial atherosclerotic disease (ICAD) is being seen as an increasingly important cause of transient ischemic attack and stroke. Attention on this disease entity has been focused as a result of two recent events: the publication of the WASID trial and the recent Food and Drug Administration approval of the Wingspan-Gateway stent system. This manuscript will review the evolution of the role of endovascular therapy in the management of ICAD. In addition, the available devices currently available for endovascular therapy will be reviewed with an eye toward peri-procedural and follow-up complication rates. PMID- 19807857 TI - Periprocedural management of patients with endovascular treatment of intracranial atherosclerotic disease. AB - Clinical outcome in endovascular therapy of intracranial atherosclerosis with stenting and angioplasty or angioplasty alone is dependent on multiple other factors beside the procedure itself. Preprocedure combined antiplatelet administration of aspirin and clopidogrel and its duration is critical. Preferably, this should be administered 5-7 days prior to the intended procedure and 90 days after the stent placement is preferred. Anticoagulation during the procedure is implemented routinely in the neurointerventional labs during intracranial intervention, with a goal of activated clotting time between 250 to 300 seconds. The preferred agent is unfractionated heparin as an intravenous bolus of 70-80 units/kg, without postprocedure reversal in most cases. Control of blood pressure intraprocedure and in the neurointensive care unit to avoid intracranial hemorrhage and hyperperfusion injury is of paramount significance. The preferred blood pressure is not well known, but lowering the blood pressure below the baseline after luminal gain with stenting would be recommended to prevent secondary injury. The use of general anesthesia versus local and monitored awake anesthesia is controversial and there are no data to support one method over another in cases of intracranial atherosclerosis interventional and endovascular therapy. If the patient is cooperative and would be able to tolerate the procedure under awake anesthesia, the latter would provide an immediate assessment of the neurological outcome and feedback from the patient. However, general anesthesia would provide the interventionalist with less movement artifact and less road mapping need and immediate and accurate assessment of the location of the microwire, catheter, and stent. PMID- 19807858 TI - Research priorities for intracranial atherosclerotic diseases. AB - The current review summarizes the characteristics of existing experimental models for intracranial atherosclerosis in rabbits, pigs, and dogs with potential implications for research. New methodologies for understanding plaque morphology, and plaque quantitation and its prognostic implications are important for risk stratification in regards to ischemic events and lesion progression. A potential treatment strategy for intracranial atherosclerotic disease may be aimed at medical therapies that induce plaque regression. The treatment with statins to stabilize and/or promote plaque regression of intracranial atherosclerotic lesions is largely inferred from data in the coronary literature. In patients with multisegmented intracranial atherosclerotic diseases with no other therapeutic option, angiogenic growth factors may represent a new venue. PMID- 19807859 TI - Resilience: one part recovery, two parts sustainability. PMID- 19807860 TI - Increasing psychological well-being and resilience by psychotherapeutic methods. AB - A specific psychotherapeutic strategy for increasing psychological well-being and resilience, well-being therapy, has been developed and validated in a number of randomized controlled trials. The findings indicate that flourishing and resilience can be promoted by specific interventions leading to a positive evaluation of one's self, a sense of continued growth and development, the belief that life is purposeful and meaningful, the possession of quality relations with others, the capacity to manage effectively one's life, and a sense of self determination. A decreased vulnerability to depression and anxiety has been demonstrated after well-being therapy in high-risk populations. There are important implications for the state/trait dichotomy in psychological well-being and for the concept of recovery in mood and anxiety disorders. PMID- 19807861 TI - Multidimensional treatment foster care as a preventive intervention to promote resiliency among youth in the child welfare system. AB - Demographic trends indicate that a growing segment of families is exposed to adversity such as poverty, drug use problems, caregiver transitions, and domestic violence. Although these risk processes and the accompanying poor outcomes for children have been well studied, little is known about why some children develop resilience in the face of such adversity, particularly when it is severe enough to invoke child welfare involvement. This paper describes a program of research involving families in the child welfare system. Using a resiliency framework, evidence from 4 randomized clinical trials that included components of the Multidimensional Treatment Foster Care program is presented. Future directions and next steps are proposed. PMID- 19807862 TI - Promoting resilience in youth from divorced families: lessons learned from experimental trials of the New Beginnings Program. AB - This article focuses on the contributions that the program of research on the New Beginnings Program (NBP) has made to understanding pathways to resilience in youth who experience parental divorce. First, the research demonstrating that divorce increases risk for mental health, physical health, and social adaptation problems is reviewed. Next, theory and research linking social environmental level and youth-level modifiable risk factors and resilience resources to youth's postdivorce adjustment are presented. The conceptual framework underlying the NBP and the risk factors and resilience resources targeted in this program are described next. The short-term and long-term results of two experimental, randomized efficacy trials of the NBP and moderators and mediators of its effects are then presented. Analyses that examine whether youth self-systems beliefs account for the links between program-induced changes in family-level resilience resources and positive long-term program on adaptation outcomes are presented and how experimental trials can be used to further theories of resilience for youth facing adversities is discussed. The final section describes directions for future research on the NBP. PMID- 19807863 TI - Predictors and parameters of resilience to loss: toward an individual differences model. AB - Although there is marked variation in how people cope with interpersonal loss, there is growing recognition that most people manage this extremely stressful experience with minimal to no impact on their daily functioning (G. A. Bonanno, 2004). What gives rise to this resilient capacity? In this paper, we provide an operational definition of resilience as a specific trajectory of psychological outcome and describe how the resilient trajectory differs from other trajectories of response to loss. We review recent data on individual differences in resilience to loss, including self-enhancing biases, repressive coping, a priori beliefs, identity continuity and complexity, dismissive attachment, positive emotions, and comfort from positive memories. We integrate these individual differences in a hypothesized model of resilience, focusing on their role in appraisal processes and the use of social resources. We conclude by considering potential cultural constraints on resilience and future research directions. PMID- 19807866 TI - Quality of reporting of clinical trials of dogs and cats and associations with treatment effects. AB - BACKGROUND: To address concerns about the quality of reporting of randomized controlled trials, and the potential for biased treatment effects in poorly reported trials, medical journals have adopted a common set of reporting guidelines, the Consolidated Standards of Reporting Trials (CONSORT) statement, to improve the reporting of randomized controlled trials. HYPOTHESIS: The reporting of clinical trials involving dogs and cats might not be ideal, and this might be associated with biased treatment effects. ANIMALS: Dogs and cats used in 100 randomly selected reports of clinical trials. METHODS: Data related to methodological quality and completeness of reporting were extracted from each trial. Associations between reporting of trial features and the proportion of positive treatment effects within trials were evaluated by generalized linear models. RESULTS: There were substantive deficiencies in reporting of key trial features. An increased proportion of positive treatment effects within a trial was associated with not reporting: the method used to generate the random allocation sequence (P < .001), the use of double blinding (P < .001), the inclusion criteria for study subjects (P = .003), baseline differences between treatment groups (P = .006), the measurement used for all outcomes (P = .002), and possible study limitations (P = .03). CONCLUSIONS AND CLINICAL IMPORTANCE: Many clinical trials involving dogs and cats in the literature do not report details related to methodological quality and aspects necessary to evaluate external validity. There is some evidence that these deficiencies are associated with treatment effects. There is a need to improve reporting of clinical trials, and guidelines, such as the CONSORT statement, can provide a valuable tool for meeting this need. PMID- 19807867 TI - Unravelling Escherichia coli dynamics close to the maximum growth temperature through heterogeneous modelling. AB - AIMS: Previous work showed that the exponential phase of Escherichia coli K12 MG1655, grown in Brain Heart Infusion broth at temperatures close to its maximum growth temperature, is disturbed. Based on plate count data, microscopic images and literature, the existence of a heat-resistant subpopulation was hypothesized. Here, this hypothesis is mathematically explored via a heterogeneous model. METHODS AND RESULTS: A heat-sensitive and a heat-resistant subpopulation are considered. A large fraction of the population is inactivated, while the remaining smaller fraction is able to resist (or adapt to) the inimical temperature and grows. A heterogeneous model that encloses a growth model (resistant population) and an inactivation model (sensitive population) is used to describe the global population dynamics. Most experimental data can be predicted when taking parameter uncertainty via Monte Carlo simulation into account. CONCLUSIONS: The heterogeneous model accurately describes disturbed growth curves at superoptimal temperatures, except for high initial cell densities. SIGNIFICANCE AND IMPACT OF THE STUDY: This study strengthens the hypothesis of the existence of a (small) heat-resistant subpopulation in typical inoculum cultures of E. coli K12 MG1655. PMID- 19807864 TI - Resilience comes of age: defining features in later adulthood. AB - Historically, resilience research has been largely the purview of developmental investigators dealing with early childhood and adolescence. This research primarily focused on at-risk children who were exposed to significant and severe life adversities (e.g., extreme poverty, parental mental illness, community violence). The study of resilience in adulthood and later life, by comparison, remains largely understudied. In this article, we describe a program of research on adulthood resilience. We begin with a selective review of the broad literature on resilience, giving emphasis to the major approaches, empirical findings, and guiding principles that characterize prior studies. We then summarize our own approach to the phenomenon of resilience and illustrate select parts of our previous and ongoing studies of older adults. Findings from this research add to the growing body of empirical evidence suggesting that resilience is a common phenomenon that emerges from the coordinated orchestration of basic human adaptive processes. PMID- 19807868 TI - The beta-lactam-sensitive D,D-carboxypeptidase activity of Pbp4 controls the L,D and D,D transpeptidation pathways in Corynebacterium jeikeium. AB - Corynebacterium jeikeium is an emerging nosocomial pathogen responsible for vascular catheters infections, prosthetic endocarditis and septicemia. The treatment of C. jeikeium infections is complicated by the multiresistance of clinical isolates to antibiotics, in particular to beta-lactams, the most broadly used class of antibiotics. To gain insight into the mechanism of beta-lactam resistance, we have determined the structure of the peptidoglycan and shown that C. jeikeium has the dual capacity to catalyse formation of cross-links generated by transpeptidases of the d,d and l,d specificities. Two ampicillin-insensitive cross-linking enzymes were identified, Ldt(Cjk1), a member of the active site cysteine l,d-transpeptidase family, and Pbp2c, a low-affinity class B penicillin binding protein (PBP). In the absence of beta-lactam, the PBPs and the l,d transpeptidase contributed to the formation of 62% and 38% of the cross-links respectively. Although Ldt(Cjk1) and Pbp2C were not inhibited by ampicillin, the participation of the l,d-transpeptidase to peptidoglycan cross-linking decreased in the presence of the drug. The specificity of Ldt(Cjk1) for acyl donors containing a tetrapeptide stem accounts for this effect of ampicillin since the essential substrate of Ldt(Cjk1) was produced by an ampicillin-sensitive d,d carboxypeptidase (Pbp4(Cjk)). Acquisition and mutational alterations of pbp2C accounted for high-level beta-lactam resistance in C. jeikeium. PMID- 19807869 TI - Differential stress-induced alterations of colonic corticotropin-releasing factor receptors in the Wistar Kyoto rat. AB - BACKGROUND A growing body of data implicates increased life stresses with the initiation, persistence and severity of symptoms associated with functional gut disorders such as irritable bowel syndrome (IBS). Activation of central and peripheral corticotropin-releasing factor (CRF) receptors is key to stress induced changes in gastrointestinal (GI) function. METHODS This study utilised immunofluorescent and Western blotting techniques to investigate colonic expression of CRF receptors in stress-sensitive Wistar Kyoto (WKY) and control Sprague Dawley (SD) rats. KEY RESULTS No intra-strain differences were observed in the numbers of colonic CRFR1 and CRFR2 positive cells. Protein expression of functional CRFR1 was found to be comparable in control proximal and distal colon samples. Sham levels of CRFR1 were also similar in the proximal colon but significantly higher in WKY distal colons (SD: 0.38 +/- 0.14, WKY: 2.06 +/- 0.52, P < 0.01). Control levels of functional CRFR2 were similar between strains but sham WKYs samples had increased CRFR2 in both the proximal (SD: 0.88 +/- 0.21, WKY: 1.8 +/- 0.18, P < 0.001) and distal (SD: 0.18 +/- 0.08, WKY: 0.94 +/- 0.32, P < 0.05) regions. Exposure to open field (OF) and colorectal distension (CRD) stressors induced decreased protein expression of CRFR1 in SD proximal colons, an effect that was blunted in WKYs. CRD stimulated decreased expression of CRFR2 in WKY rats alone. Distally, CRFR1 is decreased in WKY rats following CRD but not OF stress without any apparent changes in SD rats. CONCLUSIONS & INFERENCES This study demonstrates that psychological and physical stressors alter colonic CRF receptor expression and further support a role for local colonic CRF signalling in stress-induced changes in GI function. PMID- 19807870 TI - Cellular and molecular characterization of Phytophthora parasitica appressorium mediated penetration. AB - Data on plant pathogenic oomycetes are scarce and little is known about the early events leading to the onset of infection. The aim of this work was to analyze the penetration process of the soil-borne plant pathogen Phytophthora parasitica, which has a wide host range. Here, we performed a cytological analysis of the colonization of the first plant cell and developed an inoculation assay for characterizing the entire penetration process through cellular and molecular analyses. We showed that P. parasitica infects roots by producing a specialized structure, the appressorium. We produced the first cDNA library for the penetrating stage of a Phytophthora species and showed it to be highly enriched in pathogenicity-related sequences. These included coding sequences for many cell degrading enzymes, effectors such as RXLR-containing proteins and proteins involved in protection against plant defense responses. Characterization of the appressorium cDNA library and identification of genes overrepresented early in P. parasitica infection provided us with an unprecedented opportunity to decipher the molecular mechanisms involved in penetration of the plant cells during the initiation of infection by a soil-borne oomycete. PMID- 19807871 TI - Learned and naive natural enemy responses and the interpretation of volatile organic compounds as cues or signals. AB - In response to arthropod herbivory, plants release volatile organic compounds (VOCs), which are attractive to natural enemies. Consequently, VOCs have been interpreted as co-evolved plant-natural enemy signals. This review argues that, while these data are necessary, they are not sufficient to demonstrate a VOC plant-natural enemy signaling function. We propose that evidence that (1) plant fitness is increased as a consequence of natural enemy recruitment, and either (2A) natural enemies preferentially learn prey-induced VOCs or (2B) natural enemies respond innately to the VOCs of the prey-host plant complex, is also required. Whereas there are too few studies to rigorously test hypotheses 1 and 2A, numerous studies are available to test hypothesis 2B. Of 293 tests of natural enemy responses to VOCs, we identified only 74 that were unambiguous tests of naive natural enemies; in the remainder of the tests either natural enemies were experienced with their host in the presence of VOCs, or experience could not be ruled out. Of those 74 tests with naive natural enemies, attraction was observed in 41 and not in 33. This review demonstrates that empirical support for the hypothesized VOC plant-natural enemy signaling function is not universal and presents alternative hypotheses for VOC production. PMID- 19807872 TI - Correlated evolution of mating system and floral display traits in flowering plants and its implications for the distribution of mating system variation. AB - Reduced allocation to structures for pollinator attraction is predicted in selfing species. We explored the association between outcrossing and floral display in a broad sample of angiosperms. We used the demonstrated relationship to test for bias against selfing species in the outcrossing rate distribution, the shape of which has relevance for the stability of mixed mating. Relationships between outcrossing rate, flower size, flower number and floral display, measured as the product of flower size and number, were examined using phylogenetically independent contrasts. The distribution of floral displays among species in the outcrossing rate database was compared with that of a random sample of the same flora. The outcrossing rate was positively associated with the product of flower size and number; individually, components of display were less strongly related to outcrossing. Compared with a random sample, species in the outcrossing rate database showed a deficit of small floral display sizes. We found broad support for reduced allocation to attraction in selfing species. We suggest that covariation between mating systems and total allocation to attraction can explain the deviation from expected trade-offs between flower size and number. Our results suggest a bias against estimating outcrossing rates in the lower half of the distribution, but not specifically against highly selfing species. PMID- 19807873 TI - Action and reaction of host and pathogen during Fusarium head blight disease. AB - The Fusarium species Fusarium graminearum and Fusarium culmorum, which are responsible for Fusarium head blight (FHB) disease, reduce world-wide cereal crop yield and, as a consequence of their mycotoxin production in cereal grain, impact on both human and animal health. Their study is greatly promoted by the availability of the genomic sequence of F. graminearum and transcriptomic resources for both F. graminearum and its cereal hosts. Functional genomic, proteomic and metabolomic studies, in combination with targeted mutagenesis or transgenic studies, are unravelling the complex mechanisms involved in Fusarium infection, penetration and colonization of host tissues, and host avoidance thereof. This review illuminates and integrates emerging knowledge regarding the molecular crosstalk between Fusarium and its small-grain cereal hosts. An understanding of the complexity of the host-pathogen interactions will be instrumental in designing new efficient strategies for the control of FHB disease. PMID- 19807874 TI - 4. Cervical radicular pain. AB - Cervical radicular pain is defined as pain perceived as arising in the arm caused by irritation of a cervical spinal nerve or its roots. Approximately 1 person in 1,000 suffers from cervical radicular pain. In the absence of a gold standard, the diagnosis is based on a combination of history, clinical examination, and (potentially) complementary examination. Medical imaging may show abnormalities, but those findings may not correlate with the patient's pain. Electrophysiologic testing may be requested when nerve damage is suspected but will not provide quantitative/qualitative information about the pain. The presumed causative level may be confirmed by means of selective diagnostic blocks. Conservative treatment typically consists of medication and physical therapy. There are no studies assessing the effectiveness of different types of medication specifically in patients suffering cervical radicular pain. Cochrane reviews did not find sufficient proof of efficacy for either education or cervical traction. When conservative treatment fails, interventional treatment may be considered. For subacute cervical radicular pain, the available evidence on efficacy and safety supports a recommendation (2B+) of interlaminar cervical epidural corticosteroid administration. A recent negative randomized controlled trial of transforaminal cervical epidural corticosteroid administration, coupled with an increasing number of reports of serious adverse events, warrants a negative recommendation (2B-). Pulsed radiofrequency treatment adjacent to the cervical dorsal root ganglion is a recommended treatment for chronic cervical radicular pain (1B+). When its effect is insufficient or of short duration, conventional radiofrequency treatment is recommended (2B+). In selected patients with cervical radicular pain, refractory to other treatment options, spinal cord stimulation may be considered. This treatment should be performed in specialized centers, preferentially study related. PMID- 19807875 TI - Study on the reference values of serum lipids in children aged 3-18 years old in Beijing, China. AB - BACKGROUND: The present study was designed to explore the reference values of serum lipids in children in Beijing. METHODS: A total of 1071 healthy children from Beijing between 3 and 18 years of age were enrolled and divided into four groups: A, 3-5-year-old group; B, 6-9-year-old group; C, 10-14-year-old group; and D, 15-18-year-old group. Blood samples were obtained from the cubital veins of the participants under a fasting condition. A Hitachi 7600 Automatic Biochemistry Analyzer was used to determine the serum levels. The cut-off points of reference values were the 75th and 95th percentiles for total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) while the 25th and 5th percentiles were used for high-density lipoprotein cholesterol (HDL-C) concentrations. RESULTS: The highest 95th percentiles of TC among the four groups were 5.34 mmol/L for boys and 5.11 mmol/L for girls. Compared with those of boys, female TG levels were obviously higher in the 10-14-year-old group (P < 0.05) and HDL-C levels were significantly higher in the 15-18-year-old group (P < 0.05). The serum TC, HDL-C, and LDL-C all peaked during the 6-9-year-old period and then declined during the adolescent stage. Also, the serum TG continuously increased with a relative plateau during the 6-14-year-old period. Obese and overweight children had higher TG and LDL-C levels but lower HDL-C levels. CONCLUSION: This study showed a picture of the serum lipids distribution in Chinese children and found that their TC levels were lower than those of children in Japan, Iran, and France, but were higher than those of Italian children. PMID- 19807876 TI - Neurodevelopment in 1-year-old Japanese infants after congenital heart surgery. AB - BACKGROUND: The purpose of this study was to determine the prevalence of persistent neurodevelopmental sequelae in 1-year-old infants after open heart surgery for congenital heart disease, using the Bayley Scales of Infant Development second edition (BSID-II). A secondary objective was to confirm the applicability and usefulness of the BSID-II in Japanese infants. METHODS: Thirty nine infants who underwent repair of a ventricular septal defect before 6 months of age and 108 normal Japanese infants at 1 year of age were assessed using the BSID-II. RESULTS: In normal infants, scores on the Mental Development Index and the Psychomotor Development Index components of the BSID-II ranged from borderline retardation to very superior following a normal distribution similar to those obtained for US controls. No problems were encountered, either in translation or in following the instructions when the BSID-II was used to evaluate the 1-year-old Japanese infants. On the other hand, the mean scores on the Mental Development Index and the Psychomotor Development Index were significantly lower in Japanese patients than in normal Japanese infants, particularly for gross motor development (P < 0.001). Linear regression analysis showed that a longer intensive care unit stay was associated with impaired cognitive development at 1 year of age (P= 0.03). CONCLUSIONS: Neurodevelopmental functions in 1-year-old infants with congenital heart disease were lower than those in normal infants, especially for gross motor function using the BSID-II. One risk factor that correlated with neurodevelopmental sequelae was the length of intensive care unit stay. PMID- 19807877 TI - Anthropometric indices as predictors of the metabolic syndrome and its components in adolescents. AB - BACKGROUND: Overweight and related health problems are becoming increasingly recognized, especially in children and adolescents. For early screening, different anthropometrical measurements of obesity have been proposed to identify individuals at risk. We compared body mass index (BMI), BMI standard deviation score, waist circumference, waist-to-hip ratio (WHR), and waist/height ratio with respect to their power to predict the metabolic syndrome, its components and low grade inflammation. METHODS: A total of 79 male Caucasian German adolescents (13 17 years) were studied. All anthropometrical measurements of obesity were recorded and blood samples drawn. Predictive power was estimated using receiver operating characteristic curves, by comparing the area under the curve (AUC). RESULTS: Except for WHR, all tested anthropometrical measurements of obesity showed comparably good AUC values for correct prediction, with the highest AUC for BMI (P < 0.001, AUC = 0.885 +/- 0.039). Superior prediction power was not observed for BMI standard deviation score, waist circumference, WHR or waist/height ratio. Furthermore, BMI was the best predictor of elevated C reactive protein levels as a marker for low-grade inflammation (P < 0.001, AUC = 0.786 +/- 0.064). CONCLUSIONS: In this cross-sectional study the well-established parameter BMI was shown to have the best predictive power to identify metabolic syndrome, its components and markers for low-grade inflammation. Newly developed parameters did not provide superior values. Future longitudinal studies are needed to compare these anthropometrical markers in larger cohorts, incorporating different age groups and ethnic backgrounds. PMID- 19807878 TI - Weight disorders and associated morbidity among young adults in Israel 1990-2003. AB - BACKGROUND: The aim of this study was to evaluate the change in prevalence of weight disorders and associated morbidity among young adults. METHODS: Records of 1,021,211 subjects aged 17, who underwent a medical examination, were included. RESULTS: Among female subjects the prevalence of underweight increased steadily from 13.2% to 15.6%. No statistically significant trend was found among male subjects. Among male subjects a twofold increase in the prevalence of obesity was observed, while among female subjects there was an increase of 57%. In obese versus normal-weight male subjects the OR, adjusted for origin, education and year of recruitment, of having high blood pressure, diabetes type 2 and type 1 and hyperlipidemia were 4.1 (95%CI 3.89-4.34), 5.56 (5.09-6.07), 1.00 (0.30-3.27) and 16.07 (8.29-31.15), respectively. For obese female nominees the OR were 5.56 (95%CI 5.09-6.07), 4.42 (3.90-5.00), 3.59 (0.77-16.87) and 9.00 (4.36-18.60), respectively. CONCLUSION: A significant increase in prevalence of obesity was observed with an increased risk of having diabetes type 2, hyperlipidemia and high blood pressure. An increase in the prevalence of being underweight was observed in female subjects. Policy makers have to plan appropriate effective interventions that will promote a healthy lifestyle in adolescents and reduce obesity on the one hand, and avoid the fashion for being underweight, with its possible development to anorexia, on the other. PMID- 19807879 TI - Can pediatricians accurately identify maternal depression at well-child visits? AB - BACKGROUND: The feasibility of a two-item screening tool for maternal depression in a pediatric setting was recently reported. We assessed whether the accuracy of pediatrician recognition of maternal depression during the one-month well-child visit could be improved by an educational intervention using the two-item screening tool. METHODS: We conducted an educational intervention for pediatric residents in a suburban hospital in Tokyo, Japan, with outcome measurement before and after. Resident education included knowledge about postpartum depression and its impact on children, use of the two-item screening tool and available management strategies. Sixteen pediatric residents examined 267 mother-infant dyads during well-child visits. Residents documented the presence or absence of postpartum depressive symptoms on medical records. Depressive symptoms were also determined using the Edinburgh Postnatal Depression Scale (EPDS) survey; residents were not aware of the results. Using the EPDS as a "gold standard," improvement in sensitivity and specificity of resident recognition of maternal depressive symptoms was determined. RESULTS: The overall prevalence of postpartum depressive symptoms based on the EPDS was 15.4%. The sensitivity of resident recognition was 8% and specificity 98% before intervention, and 12% and 96% afterwards, respectively. The difference was not statistically significant. Residents indicated fear of maternal stigmatization and mothers' receptiveness to discussing depressive symptoms, as well as lack of time and skills, as major barriers to the identification of maternal depression. CONCLUSIONS: A simple educational intervention using a two-item screening tool did not improve the pediatrician's accuracy in detecting depressive symptoms in mothers. Additional strategies to address perceived barriers may be needed. PMID- 19807880 TI - C acid decarboxylases required for C photosynthesis are active in the mid-vein of the C species Arabidopsis thaliana, and are important in sugar and amino acid metabolism. AB - Cells associated with veins of petioles of C(3) tobacco possess high activities of the decarboxylase enzymes required in C(4) photosynthesis. It is not clear whether this is the case in other C(3) species, nor whether these enzymes provide precursors for specific biosynthetic pathways. Here, we investigate the activity of C(4) acid decarboxylases in the mid-vein of Arabidopsis, identify regulatory regions sufficient for this activity, and determine the impact of removing individual isoforms of each protein on mid-vein metabolite profiles. This showed that radiolabelled malate and bicarbonate fed to the xylem stream were incorporated into soluble and insoluble material in the mid-vein of Arabidopsis leaves. Compared with the leaf lamina, mid-veins possessed high activities of NADP-dependent malic enzyme (NADP-ME), NAD-dependent malic enzyme (NAD-ME) and phosphoenolpyruvate carboxykinase (PEPCK). Transcripts derived from both NAD-ME, one PCK and two of the four NADP-ME genes were detectable in these veinal cells. The promoters of each decarboxylase gene were sufficient for expression in mid veins. Analysis of insertional mutants revealed that cytosolic NADP-ME2 is responsible for 80% of NADP-ME activity in mid-veins. Removing individual decarboxylases affected the abundance of amino acids derived from pyruvate and phosphoenolpyruvate. Reducing cytosolic NADP-ME activity preferentially affected the sugar content, whereas abolishing NAD-ME affected both the amino acid and the glucosamine content of mid-veins. PMID- 19807882 TI - Characterization of SUN-domain proteins at the higher plant nuclear envelope. AB - Sad1/UNC-84 (SUN)-domain proteins are inner nuclear membrane (INM) proteins that are part of bridging complexes linking cytoskeletal elements with the nucleoskeleton, and have been shown to be conserved in non-plant systems. In this paper, we report the presence of members of this family in the plant kingdom, and investigate the two Arabidopsis SUN-domain proteins, AtSUN1 and AtSUN2. Our results indicate they contain the highly conserved C-terminal SUN domain, and share similar structural features with animal and fungal SUN-domain proteins including a functional coiled-coil domain and nuclear localization signal. Both are expressed in various tissues with AtSUN2 expression levels relatively low but upregulated in proliferating tissues. Further, we found AtSUN1 and AtSUN2 expressed as fluorescent protein fusions, to localize to and show low mobility in the nuclear envelope (NE), particularly in the INM. Deletion of various functional domains including the N terminus and coiled-coil domain affect the localization and increase the mobility of AtSUN1 and AtSUN2. Finally, we present evidence that AtSUN1 and AtSUN2 are present as homomers and heteromers in vivo, and that the coiled-coil domains are required for this. The study provides evidence suggesting the existence of cytoskeletal-nucleoskeletal bridging complexes at the plant NE. PMID- 19807881 TI - Inactivation of the CTD phosphatase-like gene OsCPL1 enhances the development of the abscission layer and seed shattering in rice. AB - Although susceptibility to seed shattering causes severe yield loss during cereal crop harvest, it is an adaptive trait for seed dispersal in wild plants. We previously identified a recessive shattering locus, sh-h, from the rice shattering mutant line Hsh that carries an enhanced abscission layer. Here, we further mapped sh-h to a 34-kb region on chromosome 7 by analyzing 240 F(2) plants and five F(3) lines from the cross between Hsh and Blue&Gundil. Hsh had a point mutation at the 3' splice site of the seventh intron within LOC_Os07g10690, causing a 15-bp deletion of its mRNA as a result of altered splicing. Two transferred DNA (T-DNA) insertion mutants and one point mutant exhibited the enhanced shattering phenotype, confirming that LOC_Os07g10690 is indeed the sh-h gene. RNA interference (RNAi) transgenic lines with suppressed expression of this gene exhibited greater shattering. This gene, which encodes a protein containing a conserved carboxy-terminal domain (CTD) phosphatase domain, was named Oryza sativa CTD phosphatase-like 1 (OsCPL1). Subcellular localization and biochemical analysis revealed that the OsCPL1 protein is a nuclear phosphatase, a common characteristic of metazoan CTD phosphatases involved in cell differentiation. These results demonstrate that OsCPL1 represses differentiation of the abscission layer during panicle development. PMID- 19807883 TI - Callers' perceptions of receiving advice via a medical care help line. AB - BACKGROUND: Telephone consultations with specially trained nurses are becoming an increasingly common form of care. AIM: To describe patients' perceptions of receiving advice via a medical care help line. RESULTS: The patients perceived the help line as a professional, reliable and easily accessible asset in everyday life, that self-care is promoted through personal advice and that the help line is a partner with whom one can discuss reflections and feelings. It is also a kind of 'back up'. The advice service is perceived as satisfactory when the nurse is calm, friendly, confirming and shows respect. Compliance and acceptance are enhanced when patients feel involved in the decision-making process. The fact that the service is easily accessible is perceived as simple and time saving. CONCLUSIONS: Seen from the patient's perspective, the telephone contact with the help line is a simple, easily accessible and secure alternative that is appreciated and used. Caring encounters gave rise to feelings, influenced by the agreement between one's own needs and expectations, the encounter between human beings and the care provided. More in-depth studies is needed focusing on the patient's perspective and characterizes of the caring encounter over the telephone. RELEVANCE TO CLINICAL NURSING: Improved understanding of the patient's perspective on the care provided leads to increased staff satisfaction and motivation. Care encounters over the phone are common today for nurses and their well-being at work is beneficial for both themselves and the care they provide and thereby for the patients. PMID- 19807884 TI - Spinal muscular atrophy: the challenges of 'doing the right thing'. PMID- 19807885 TI - Impact of normal saline infusion on postoperative metabolic acidosis. AB - BACKGROUND: Standard base excess (SBE) is an important parameter for guiding fluid management in postoperative metabolic acidosis. However, individual SBE components, notably the chloride effect (Cl(eff)), provide valuable additional information. Cl(eff) is the deviation of the strong ion difference (SID) from normal caused by chloride loss or increase and represents the effect on SBE of an abnormal chloride-sodium ratio. Many centers use normal saline (NS) for intravascular volume therapy. In this study, we examined the impact of NS infusion on SBE and its chloride-driven component (Cl(eff)) in postoperative children. METHODS: The study was conducted in 119 children who underwent post heart surgery in a Swiss pediatric intensive care unit. The 72-h postoperative course was divided into six observation periods, during which NS input and its impact on SBE and Cl(eff) were measured per period in each patient, and the results compared between patients infused and not infused with NS during each period. RESULTS: Normal saline was infused in 168/625 observation periods if indicated by volume deficit. Postoperative metabolic acidosis and the acidifying Cl(eff) were aggravated in the first 12 postoperative hours. Over the 72 h, NS infusion simultaneously lowered SBE by -0.06 mm x ml(-1) x kg(-1) body weight infused and Cl(eff) by -0.07 mm. CONCLUSIONS: Implementing serial Cl(eff) assessment could improve postoperative management by disclosing or excluding hyperchloremia as a cause of acidosis undetectable from SBE alone. Calculating the chloride-driven acidifying side effect of NS infusion using Cl(eff) improves the interpretation of SBE values and can optimize fluid management in postoperative metabolic acidosis. PMID- 19807886 TI - Parent-assisted or nurse-assisted epidural analgesia: is this feasible in pediatric patients? AB - AIM: The aim of this study was to assess the feasibility of parent-assisted or nurse-assisted epidural analgesia (PNEA) for control of postoperative pain in a pediatric surgical population. METHODS: After the institutional review board (IRB) approval was obtained, an analysis of our pain treatment services database of pediatric surgical patients with epidural catheters in whom the parent and/or nurse were empowered to activate the epidural demand-dose button was evaluated. RESULTS: Over a 10 -year period between 1999 and 2008, 128 procedures in 126 patients were provided parent or nurse assistance of the epidural demand dose. Satisfactory analgesia was obtained in 86% of patients with no or minor adjustments in PNEA parameters. Fourteen percent of patients were converted to intravenous patient-controlled analgesia (PCA) for inadequate analgesia (7%) or side effects (7%). None of the patients in this cohort required treatment for respiratory depression or excessive sedation. CONCLUSIONS: Parent-assisted or nurse-assisted epidural analgesia can be safely administered to children undergoing surgery who are physically or cognitively unable or unwilling to self activate a demand dose. Additional studies are needed to compare the efficacy of PNEA with other modalities for postoperative pain control in children. PMID- 19807887 TI - Epinephrine test dose in children: is it interpretable on ECG monitor? AB - BACKGROUND: Epidural and other regional blocks are performed in children under general anesthesia; the response to a 'test dose' may be altered during administration of general anesthetics. Limited data is available describing changes in electrocardiogram, blood pressure and heart rate (HR) following unintentional intravascular injection of a lidocaine-epinephrine-containing test dose, under sevoflurane anesthesia in children. METHODS: Sixty-eight children undergoing elective surgeries under sevoflurane anesthesia were administered 0.1 ml x kg(-1) of 1% lidocaine with epinephrine 0.5 microg x kg(-1) or normal saline intravenously, to simulate an accidental intravascular test dose. T-wave changes in lead II on the anesthesia monitor and on a printed ECG were noted over the initial 1 min as well as changes in HR and systolic blood pressure (SBP) over an initial 3 min period. RESULTS: Following injection of lidocaine-epinephrine, a significant increase in T-wave amplitude in lead II was noted in 91% of children on the ECG monitor and in 94% of children on the ECG printout of the same lead. In 64% of children, an increase in HR of > or =10 b x min(-1) and in 76% of children an increase in SBP of > or =15 mmHg was noted. CONCLUSION: An increase in T-wave amplitude can easily be detected by carefully observing the ECG monitor or an ECG printout within a minute following the accidental i.v. administration of 0.1 ml x kg(-1) of 1% lidocaine-epinephrine (0.5 microg x kg(-1)) regional anesthetic test dose in children under sevoflurane anesthesia. PMID- 19807888 TI - Anesthesia in Brown-Vialetto-Van Laere syndrome. PMID- 19807889 TI - Fibreoptic intubation through the LMA in children. PMID- 19807890 TI - Laryngeal mask airway-aided fiberoptic tracheal intubation in infant--a modified technique. PMID- 19807891 TI - Use of an adult fiberoptic bronchoscope to facilitate endotracheal intubation in children with difficult airways. PMID- 19807892 TI - Use of the Glidescope video laryngoscope. PMID- 19807893 TI - Use of an extraoral method to fix endotracheal tube during craniomaxillofacial surgery. PMID- 19807894 TI - Epidural catheter occlusion: disease-related or technique-related? PMID- 19807895 TI - Ganglion impar block for management of chronic coccydynia in an adolescent. PMID- 19807896 TI - Is there an association between ACTN3 R577X polymorphism and muscle power phenotypes in young, non-athletic adults? AB - We investigated the association between ACTN3 R577X polymorphism and jumping (vertical squat and counter-movement jump tests) and sprint ability (30 m dash) in non-athletic, healthy young adults [N=284 (217 male), mean (SD) age: 21 (2) years]. We analyzed the differences in the study phenotypes among ACTN3 R577X genotypes by one-way analysis of covariance before and after adjusting for sex, age, weight and height (confounders). We also compared the genotype and allele frequencies between those with the best and worst results in the aforementioned tests (>=90th vs <90th of the sex-specific percentile, respectively). We used logistic regression to calculate the odds ratio (OR) for having the best performance. We did not observe a significant association between ACTN3 R577X genotypes and the study phenotypes before and after adjusting for potential confounders, nor after analyzing males and females separately. We did not observe significant differences in genotype frequencies between those with the best or the worst performance. The OR for an individual with the RR genotype to be in the top 10 percentile was <1.00 for jump tests and <1.015 for sprint tests (all P>0.05). In summary, alpha-actinin-3 deficiency does not negatively influence the ability to generate explosive leg muscle power in a young non-athletic population. PMID- 19807898 TI - Hyperbaric oxygen therapy in liver transplantation; is its use limited to the management of hepatic artery thrombosis? PMID- 19807899 TI - Shifting priorities in vector biology to improve control of vector-borne disease. AB - Vector control remains the primary measure available to prevent pathogen transmission for the most devastating vector-borne diseases (VBDs): malaria, dengue, trypanosomiasis, filariasis, leishmaniasis, and Chagas disease. Current control strategies, however, are proving insufficient and the remarkable advances in the molecular biology of disease vectors over the last two decades have yet to result in tangible tools that effectively reduce VBD incidence. Here we argue that vector biologists must fundamentally shift their approach to VBD research. We propose an agenda highlighting the most critical avenues to improve the effectiveness of vector control. Research priorities must be diversified to support simultaneous development of multiple, alternative control strategies. Knowledge across relevant diseases and disciplines should be better integrated and disease prevention efforts extended beyond the academic sector to involve private industry, ministries of health, and local communities. To obtain information of more immediate significance to public health, the research focus must shift from laboratory models to natural pathogen-transmission systems. Identification and characterization of heterogeneities inherent to VBD systems should be prioritized to allow development of local, adaptive control strategies that efficiently make use of limited resources. Importantly, increased involvement of disease-endemic country (DEC) scientists, institutes, and communities will be key to enhance and sustain the fight against VBD. PMID- 19807897 TI - Vitamin D and human skeletal muscle. AB - Vitamin D deficiency is an increasingly described phenomenon worldwide, with well known impacts on calcium metabolism and bone health. Vitamin D has also been associated with chronic health problems such as bowel and colonic cancer, arthritis, diabetes and cardiovascular disease. In recent decades, there has been increased awareness of the impact of vitamin D on muscle morphology and function, but this is not well recognized in the Sports Medicine literature. In the early 20th century, athletes and coaches felt that ultraviolet rays had a positive impact on athletic performance, and increasingly, evidence is accumulating to support this view. Both cross-sectional and longitudinal studies allude to a functional role for vitamin D in muscle and more recently the discovery of the vitamin D receptor in muscle tissue provides a mechanistic understanding of the function of vitamin D within muscle. The identification of broad genomic and non genomic roles for vitamin D within skeletal muscle has highlighted the potential impact vitamin D deficiency may have on both under-performance and the risk of injury in athletes. This review describes the current understanding of the role vitamin D plays within skeletal muscle tissue. PMID- 19807900 TI - Cerebrospinal fluid B lymphocyte identification for diagnosis and follow-up in human African trypanosomiasis in the field. AB - OBJECTIVES: In human African trypanosomiasis (HAT, sleeping sickness), staging of disease and treatment follow-up relies on white cell count in the cerebrospinal fluid (CSF). As B lymphocytes (CD19 positive cells) are not found in the CSF of healthy individuals but occur in neurological disorders such as multiple sclerosis, B lymphocyte count may be useful for field diagnosis/staging and therapeutic follow-up in HAT. METHODS: Seventy-one HAT patients were diagnosed and 50 were followed-up 6-24 months after treatment. White cell counts were used for conventional staging (stage 1, < or =5 cells/microl CSF, n = 42; stage 2, > or =20 cells/microl, n = 16) and intermediate stage (6-19 cells/microl, n = 13). Slides containing 1 microl of CSF mixed with Dynabeads CD19 pan B were examined microscopically to detect B cell rosettes (bound to at least four beads). RESULTS: Stage 1 patients exhibited zero (n = 37) or one CSF rosette/microl (n = 5), contrary to most stage 2 patients (14/16: > or =2 rosettes/microl). Intermediate stage patients expressed 0 (n = 9), 1 (n = 3) or 2 (n = 1) rosettes/microl of CSF. During follow-up, rosette counts correlated with white cell count staging but were much easier to read. CONCLUSION: B cell rosettes being easily detected in the CSF in field conditions may be proposed to replace white cell count for defining HAT stages 1 and 2 and limit uncertainty in treatment decision in patients with intermediate stage. PMID- 19807901 TI - Community-based validation of assessment of newborn illnesses by trained community health workers in Sylhet district of Bangladesh. AB - OBJECTIVES: To validate trained community health workers' recognition of signs and symptoms of newborn illnesses and classification of illnesses using a clinical algorithm during routine home visits in rural Bangladesh. METHODS: Between August 2005 and May 2006, 288 newborns were assessed independently by a community health worker and a study physician. Based on a 20-sign algorithm, sick neonates were classified as having very severe disease, possible very severe disease or no disease. The physician's assessment was considered as the gold standard. RESULTS: Community health workers correctly classified very severe disease in newborns with a sensitivity of 91%, specificity of 95% and kappa value of 0.85 (P < 0.001). Community health workers' recognition showed a sensitivity of more than 60% and a specificity of 97-100% for almost all signs and symptoms. CONCLUSION: Community health workers with minimal training can use a diagnostic algorithm to identify severely ill newborns with high validity. PMID- 19807902 TI - Paradigm shift for tuberculosis control in high prevalence countries. PMID- 19807903 TI - Good research practices for measuring drug costs in cost-effectiveness analyses: Medicare, Medicaid and other US government payers perspectives: the ISPOR Drug Cost Task Force report--Part IV. AB - OBJECTIVES: Public programs finance a large share of the US pharmaceutical expenditures. To date, there are not guidelines for estimating the cost of drugs financed by US public programs. The objective of this study was to provide standards for estimating the cost of drugs financed by US public programs for utilization in pharmacoeconomic evaluations. METHODS: This report was prepared by the ISPOR Task Force on Good Research Practices-Use of Drug Costs for Cost Effectiveness Analysis Medicare, Medicaid, and other US Government Payers Subgroup. The Subgroup was convened to assess the methodological and practical issues confronted by researchers when estimating the cost of drugs financed by US public programs, and to propose standards for more transparent, accurate and consistent costing methods. RESULTS: The Subgroup proposed these recommendations: 1) researchers must consider regulation requirements that affect the drug cost paid by public programs; 2) drug cost must represent the actual acquisition cost, incorporating any rebates or discounts; 3) transparency with respect to cost inputs must be ensured; 4) inclusion of the public program's perspective is recommended; 5) high cost drugs require special attention, particularly when drugs represent a significant proportion of health-care expenditures for a specific disease; and 6) because of variations across public programs, sensitivity analyses for actual acquisition cost, real-world adherence, and generics availability are warranted. Specific recommendations also were proposed for the Medicare and Medicaid programs. CONCLUSIONS: As pharmacoeconomic evaluations for coverage decisions made by US public programs grows, the need for precise and consistent estimation of drug costs is warranted. Application of the proposed recommendations will allow researchers to include accurate and unbiased cost estimates in pharmacoeconomic evaluations. PMID- 19807904 TI - Gender differences in presentation rates, deferrals and return behaviour among Norwegian blood donors. AB - BACKGROUND AND OBJECTIVES: Women are under-represented among long-term blood donors. Reasons for this were sought in the donor pool of the Blood Bank of Oslo, Norway, which comprises only voluntary, non-remunerated donors and has a high degree of stability. METHODS: Three sources of data were analyzed: (1) the subsequent six-year donation patterns of 17 812 donors who donated at least once in 1999; (2) reasons for pre-donation deferral of 484 prospect donors in 2004; (3) reasons for deferrals and absence during a 6.5-year period, retrieved from a follow-up study of 1029 donors who took part in a questionnaire study on motivation for blood donation in 2000. RESULTS: Women were over-represented among first-time donors and under-represented among regular donors. Women below the age of 45 years in 1999 were less likely than men to donate regularly throughout the 6-year study period, whereas the donation behaviour of women and men above 45 years of age was similar. Young (18-29 years) female prospect donors were more frequently deferred at first-time donation than males. In the 6.5-year follow-up study, pregnancy was the most frequently reported cause of absence from or termination of donation, and was reported by 32% of the female respondents that were 45 years or younger. Among the donors that reported having been pregnant, 42% stated to have resumed donation and < 4% stated that they no longer were blood donors. Reported termination of donation by female donors was associated with reported practical obstacles and discomfort related to donation, but not with loss of motivation. CONCLUSION: Most of the gender differences in donation patterns could be ascribed to absence because of pregnancy and lactation. Practical problems and discomfort during donation were important reasons why women reported to have stopped donation. Current deferral criteria pose problems for the recruitment and retention especially of young women. PMID- 19807905 TI - A theoretical framework to describe communication processes during medical disability assessment interviews. AB - BACKGROUND: Research in different fields of medicine suggests that communication is important in physician-patient encounters and influences satisfaction with these encounters. It is argued that this also applies to the non-curative tasks that physicians perform, such as sickness certification and medical disability assessments. However, there is no conceptualised theoretical framework that can be used to describe intentions with regard to communication behaviour, communication behaviour itself, and satisfaction with communication behaviour in a medical disability assessment context. OBJECTIVE: The objective of this paper is to describe the conceptualization of a model for the communication behaviour of physicians performing medical disability assessments in a social insurance context and of their claimants, in face-to-face encounters during medical disability assessment interviews and the preparation thereof. CONCEPTUALIzATION: The behavioural model, based on the Theory of Planned Behaviour (TPB), is conceptualised for the communication behaviour of social insurance physicians and claimants separately, but also combined during the assessment interview. Other important concepts in the model are the evaluation of communication behaviour (satisfaction), intentions, attitudes, skills, and barriers for communication. CONCLUSION: The conceptualization of the TPB-based behavioural model will help to provide insight into the communication behaviour of social insurance physicians and claimants during disability assessment interviews. After empirical testing of the relationships in the model, it can be used in other studies to obtain more insight into communication behaviour in non-curative medicine, and it could help social insurance physicians to adapt their communication behaviour to their task when performing disability assessments. PMID- 19807906 TI - Detection of multiple quantitative trait loci and their pleiotropic effects in outbred pig populations. AB - BACKGROUND: Simultaneous detection of multiple QTLs (quantitative trait loci) may allow more accurate estimation of genetic effects. We have analyzed outbred commercial pig populations with different single and multiple models to clarify their genetic properties and in addition, we have investigated pleiotropy among growth and obesity traits based on allelic correlation within a gamete. METHODS: Three closed populations, (A) 427 individuals from a Yorkshire and Large White synthetic breed, (B) 547 Large White individuals and (C) 531 Large White individuals, were analyzed using a variance component method with one-QTL and two QTL models. Six markers on chromosome 4 and five to seven markers on chromosome 7 were used. RESULTS: Population A displayed a high test statistic for the fat trait when applying the two-QTL model with two positions on two chromosomes. The estimated heritabilities for polygenic effects and for the first and second QTL were 19%, 17% and 21%, respectively. The high correlation of the estimated allelic effect on the same gamete and QTL test statistics suggested that the two separate QTL which were detected on different chromosomes both have pleiotropic effects on the two fat traits. Analysis of population B using the one-QTL model for three fat traits found a similar peak position on chromosome 7. Allelic effects of three fat traits from the same gamete were highly correlated suggesting the presence of a pleiotropic QTL. In population C, three growth traits also displayed similar peak positions on chromosome 7 and allelic effects from the same gamete were correlated. CONCLUSION: Detection of the second QTL in a model reduced the polygenic heritability and should improve accuracy of estimated heritabilities for both QTLs. PMID- 19807908 TI - Initial psychological responses to Influenza A, H1N1 ("Swine flu"). AB - BACKGROUND: The outbreak of the pandemic flu, Influenza A H1N1 (Swine Flu) in early 2009, provided a major challenge to health services around the world. Previous pandemics have led to stockpiling of goods, the victimisation of particular population groups, and the cancellation of travel and the boycotting of particular foods (e.g. pork). We examined initial behavioural and attitudinal responses towards Influenza A, H1N1 ("Swine flu") in the six days following the WHO pandemic alert level 5, and regional differences in these responses. METHODS: 328 respondents completed a cross-sectional Internet or paper-based questionnaire study in Malaysia (N = 180) or Europe (N = 148). Measures assessed changes in transport usage, purchase of preparatory goods for a pandemic, perceived risk groups, indicators of anxiety, assessed estimated mortality rates for seasonal flu, effectiveness of seasonal flu vaccination, and changes in pork consumption RESULTS: 26% of the respondents were 'very concerned' about being a flu victim (42% Malaysians, 5% Europeans, p < .001). 36% reported reduced public transport use (48% Malaysia, 22% Europe, p < .001), 39% flight cancellations (56% Malaysia, 17% Europe, p < .001). 8% had purchased preparatory materials (e.g. face masks: 8% Malaysia, 7% Europe), 41% Malaysia (15% Europe) intended to do so (p < .001). 63% of Europeans, 19% of Malaysians had discussed the pandemic with friends (p < .001). Groups seen as at 'high risk' of infection included the immune compromised (mentioned by 87% respondents), pig farmers (70%), elderly (57%), prostitutes/highly sexually active (53%), and the homeless (53%). In data collected only in Europe, 64% greatly underestimated the mortality rates of seasonal flu, 26% believed seasonal flu vaccination gave protection against swine flu. 7% had reduced/stopped eating pork. 3% had purchased anti-viral drugs for use at home, while 32% intended to do so if the pandemic worsened. CONCLUSION: Initial responses to Influenza A show large regional differences in anxiety, with Malaysians more anxious and more likely to reduce travel and to buy masks and food. Discussions with family and friends may reinforce existing anxiety levels. Particular groups (homosexuals, prostitutes, the homeless) are perceived as at greater risk, potentially leading to increased prejudice during a pandemic. Europeans underestimated mortality of seasonal flu, and require more information about the protection given by seasonal flu inoculation. PMID- 19807907 TI - Therapeutic targets and limits of minocycline neuroprotection in experimental ischemic stroke. AB - BACKGROUND: Minocycline, a second-generation tetracycline with anti-inflammatory and anti-apoptotic properties, has been shown to promote therapeutic benefits in experimental stroke. However, equally compelling evidence demonstrates that the drug exerts variable and even detrimental effects in many neurological disease models. Assessment of the mechanism underlying minocycline neuroprotection should clarify the drug's clinical value in acute stroke setting. RESULTS: Here, we demonstrate that minocycline attenuates both in vitro (oxygen glucose deprivation) and in vivo (middle cerebral artery occlusion) experimentally induced ischemic deficits by direct inhibition of apoptotic-like neuronal cell death involving the anti-apoptotic Bcl-2/cytochrome c pathway. Such anti apoptotic effect of minocycline is seen in neurons, but not apparent in astrocytes. Our data further indicate that the neuroprotection is dose-dependent, in that only low dose minocycline inhibits neuronal cell death cascades at the acute stroke phase, whereas the high dose exacerbates the ischemic injury. CONCLUSION: The present study advises our community to proceed with caution to use the minimally invasive intravenous delivery of low dose minocycline in order to afford neuroprotection that is safe for stroke. PMID- 19807909 TI - Fluid-structure interaction of a patient-specific abdominal aortic aneurysm treated with an endovascular stent-graft. AB - BACKGROUND: Abdominal aortic aneurysms (AAA) are local dilatations of the infrarenal aorta. If left untreated they may rupture and lead to death. One form of treatment is the minimally invasive insertion of a stent-graft into the aneurysm. Despite this effective treatment aneurysms may occasionally continue to expand and this may eventually result in post-operative rupture of the aneurysm. Fluid-structure interaction (FSI) is a particularly useful tool for investigating aneurysm biomechanics as both the wall stresses and fluid forces can be examined. METHODS: Pre-op, Post-op and Follow-up models were reconstructed from CT scans of a single patient and FSI simulations were performed on each model. The FSI approach involved coupling Abaqus and Fluent via a third-party software - MpCCI. Aneurysm wall stress and compliance were investigated as well as the drag force acting on the stent-graft. RESULTS: Aneurysm wall stress was reduced from 0.38 MPa before surgery to a value of 0.03 MPa after insertion of the stent-graft. Higher stresses were seen in the aneurysm neck and iliac legs post-operatively. The compliance of the aneurysm was also reduced post-operatively. The peak Post op axial drag force was found to be 4.85 N. This increased to 6.37 N in the Follow-up model. CONCLUSION: In a patient-specific case peak aneurysm wall stress was reduced by 92%. Such a reduction in aneurysm wall stress may lead to shrinkage of the aneurysm over time. Hence, post-operative stress patterns may help in determining the likelihood of aneurysm shrinkage post EVAR. Post operative remodelling of the aneurysm may lead to increased drag forces. PMID- 19807910 TI - Strong association between pseudogenization mechanisms and gene sequence length. AB - Pseudogenes arise from the decay of gene copies following either RNA-mediated duplication (processed pseudogenes) or DNA-mediated duplication (nonprocessed pseudogenes). Here, we show that long protein-coding genes tend to produce more nonprocessed pseudogenes than short genes, whereas the opposite is true for processed pseudogenes. Protein-coding genes longer than 3000 bp are 6 times more likely to produce nonprocessed pseudogenes than processed ones. REVIEWERS: This article was reviewed by Dr. Dan Graur and Dr. Craig Nelson (nominated by Dr. J Peter Gogarten). PMID- 19807911 TI - Draft genome sequence of the Daphnia pathogen Octosporea bayeri: insights into the gene content of a large microsporidian genome and a model for host-parasite interactions. AB - BACKGROUND: The highly compacted 2.9-Mb genome of Encephalitozoon cuniculi placed the microsporidia in the spotlight, encoding a mere 2,000 proteins and a highly reduced suite of biochemical pathways. This extreme level of reduction is not universal across the microsporidia, with genomes known to vary up to sixfold in size, suggesting that some genomes may harbor a gene content that is not as reduced as that of Enc. cuniculi. In this study, we present an in-depth survey of the large genome of Octosporea bayeri, a pathogen of Daphnia magna, with an estimated genome size of 24 Mb, in order to shed light on the organization and content of a large microsporidian genome. RESULTS: Using Illumina sequencing, 898 Mb of O. bayeri genome sequence was generated, resulting in 13.3 Mb of unique sequence. We annotated a total of 2,174 genes, of which 893 encodes proteins with assigned function. The gene density of the O. bayeri genome is very low on average, but also highly uneven, so gene-dense regions also occur. The data presented here suggest that the O. bayeri proteome is well represented in this analysis and is more complex that that of Enc. cuniculi. Functional annotation of O. bayeri proteins suggests that this species might be less biochemically dependent on its host for its metabolism than its more reduced relatives. CONCLUSIONS: The combination of the data presented here, together with the imminent annotated genome of Daphnia magna, will provide a wealth of genetic and genomic tools to study host-parasite interactions in an interesting model for pathogenesis. PMID- 19807912 TI - Clostridium septicum sepsis and colorectal cancer - a reminder. AB - BACKGROUND: Spontaneous clostridium septicum infections are rare and are associated with a high mortality. Association of clostridium infection with colorectal malignancies have been previously reported and most cases are described in tumours of the ascending colon. We report our experience of clostridium septicum infection in the presence of tumour perforation in a series of two patients as a reminder of its association with sepsis in the presence of colorectal malignancy. CASE PRESENTATION: We isolated clostridium septicum infection in a series of two patients admitted as emergencies. One patient was found to have a perforated caecal tumour intraoperatively whilst the other had a perforated rectal tumour. The clinical outcome and management of each case are reported and underlying reasons for variations in outcome are discussed. CONCLUSION: Although uncommon, the possibility of clostridium septicum sepsis should be borne in mind in patients who present with underlying malignancy and have sepsis. The cumulative effect of sepsis and malignant perforation is associated with a high morbidity and mortality. Awareness and early diagnosis of clostridium septicum may improve the prognosis of what is usually regarded as a fatal infection. PMID- 19807913 TI - Defining the chromatin signature of inducible genes in T cells. AB - BACKGROUND: Specific chromatin characteristics, especially the modification status of the core histone proteins, are associated with active and inactive genes. There is growing evidence that genes that respond to environmental or developmental signals may possess distinct chromatin marks. Using a T cell model and both genome-wide and gene-focused approaches, we examined the chromatin characteristics of genes that respond to T cell activation. RESULTS: To facilitate comparison of genes with similar basal expression levels, we used expression-profiling data to bin genes according to their basal expression levels. We found that inducible genes in the lower basal expression bins, especially rapidly induced primary response genes, were more likely than their non-responsive counterparts to display the histone modifications of active genes, have RNA polymerase II (Pol II) at their promoters and show evidence of ongoing basal elongation. There was little or no evidence for the presence of active chromatin marks in the absence of promoter Pol II on these inducible genes. In addition, we identified a subgroup of genes with active promoter chromatin marks and promoter Pol II but no evidence of elongation. Following T cell activation, we find little evidence for a major shift in the active chromatin signature around inducible gene promoters but many genes recruit more Pol II and show increased evidence of elongation. CONCLUSIONS: These results suggest that the majority of inducible genes are primed for activation by having an active chromatin signature and promoter Pol II with or without ongoing elongation. PMID- 19807914 TI - miRNAs in lung cancer - studying complex fingerprints in patient's blood cells by microarray experiments. AB - BACKGROUND: Deregulated miRNAs are found in cancer cells and recently in blood cells of cancer patients. Due to their inherent stability miRNAs may offer themselves for blood based tumor diagnosis. Here we addressed the question whether there is a sufficient number of miRNAs deregulated in blood cells of cancer patients to be able to distinguish between cancer patients and controls. METHODS: We synthesized 866 human miRNAs and miRNA star sequences as annotated in the Sanger miRBase onto a microarray designed by febit biomed gmbh. Using the fully automated Geniom Real Time Analyzer platform, we analyzed the miRNA expression in 17 blood cell samples of patients with non-small cell lung carcinomas (NSCLC) and in 19 blood samples of healthy controls. RESULTS: Using t test, we detected 27 miRNAs significantly deregulated in blood cells of lung cancer patients as compared to the controls. Some of these miRNAs were validated using qRT-PCR. To estimate the value of each deregulated miRNA, we grouped all miRNAs according to their diagnostic information that was measured by Mutual Information. Using a subset of 24 miRNAs, a radial basis function Support Vector Machine allowed for discriminating between blood cell samples of tumor patients and controls with an accuracy of 95.4% [94.9%-95.9%], a specificity of 98.1% [97.3%-98.8%], and a sensitivity of 92.5% [91.8%-92.5%]. CONCLUSION: Our findings support the idea that neoplasia may lead to a deregulation of miRNA expression in blood cells of cancer patients compared to blood cells of healthy individuals. Furthermore, we provide evidence that miRNA patterns can be used to detect human cancers from blood cells. PMID- 19807915 TI - Recurrence in oral and pharyngeal cancer is associated with quantitative MGMT promoter methylation. AB - BACKGROUND: Biomarkers that predict clinical response, tumor recurrence or patient survival are severely lacking for most cancers, particularly for oral and pharyngeal cancer. This study examines whether gene-promoter methylation of tumor DNA correlates with survival and recurrence rates in a population of patients with oral or pharyngeal cancer. METHODS: The promoter methylation status of the DNA repair gene MGMT and the tumor suppressor genes CDKN2A and RASSF1 were evaluated by methylation-specific PCR in 88 primary oral and pharyngeal tumors and correlated with survival and tumor recurrence. Quantitative MGMT methylation was also assessed. RESULTS: 29.6% of the tumors presented with MGMT methylation, 11.5% with CDKN2A methylation and 12.1% with RASSF1 methylation. MGMT promoter methylation was significantly associated with poorer overall and disease-free survival. No differences in methylation status of MGMT and RASSF1 with HPV infection, smoking or drinking habits were observed. A significant inverse trend with the amount of MGMT methylation and overall and disease-free survival was observed (ptrend = 0.002 and 0.001 respectively). CONCLUSION: These results implicate MGMT promoter methylation as a possible biomarker for oral and pharyngeal cancer prognosis. The critical role of MGMT in DNA repair suggests that defective DNA repair may be correlative in the observed association between MGMT promoter methylation and tumor recurrence. Follow-up studies should include further quantitative MSP-PCR measurement, global methylation profiling and detailed analysis of downstream DNA repair genes regulated by promoter methylation. PMID- 19807916 TI - Molecular cloning and characterization of the porcine prostaglandin transporter (SLCO2A1): evaluation of its role in F4 mediated neonatal diarrhoea. AB - BACKGROUND: Because prostaglandins are involved in many (patho)physiological processes, SLCO2A1 was already characterized in several species in an attempt to unravel specific processes/deficiencies. Here, we describe the molecular cloning and characterization of the porcine ortholog in order to evaluate its possible involvement in F4 enterotoxigenic E. coli mediated neonatal diarrhoea, based on a positional candidate gene approach study. RESULTS: Porcine SLCO2A1 is organized in 14 exons, containing an open reading frame of 1935 bp, encoding a 12 transmembrane organic anion cell surface transporter of 644 aa. The -388 to -5 upstream region comprises a (CpG)48 island containing a number of conserved promoter elements, including a TATA box. A potential alternative promoter region was found in the conserved -973 to -700 upstream region. No consensus polyadenylation signal was discovered in the 3' UTR. Repeat sequences were found in 15% of all the non coding sequences.As expected for a multifunctional protein, a wide tissue distribution was observed. mRNA expression was found in the adrenal gland, bladder, caecum, colon (centripetal coil/centrifugal coil), diaphragm, duodenum, gallbladder, heart, ileum, jejunum, kidney, liver, longissimus dorsi muscle, lung, lymph node, mesenterium, rectum, spleen, stomach, tongue and ureter, but not in the aorta, oesophagus and pancreas.The promoter region and the exons (including the splice sites) of SLCO2A1 were resequenced in 5 F4ab/ac receptor positive and 5 F4ab/ac receptor negative pigs. Two silent and 2 missense (both S --> L at position 360 and 633) mutations were found, but none was associated with the F4ab/ac receptor phenotype. In addition, no phenotype associated differential mRNA expression or alternative/abberant splicing/polyadenylation was found in the jejunum. CONCLUSION: The molecular cloning and characterization of porcine SLCO2A1 not only contributes to the already existing knowledge about the transporter in general, but enables studies on porcine prostaglandin related processes/deficiencies as patient and/or model. Here we examined its possible involvement as receptor in F4 enterotoxigenic E. coli mediated neonatal diarrhoea. Because no phenotype associated differences could be found in the gene sequence nor in its jejunal transcription profile of F4ab/ac receptor positive/negative pigs, SLCO2A1 can most likely be excluded as receptor for F4 bacteria. PMID- 19807917 TI - FRAGMATIC: a randomised phase III clinical trial investigating the effect of fragmin added to standard therapy in patients with lung cancer. AB - BACKGROUND: Venous thromboembolism (VTE) occurs when blood clots in the leg, pelvic or other deep vein (deep vein thrombosis) with or without transport of the thrombus into the pulmonary arterial circulation (pulmonary embolus). VTE is common in patients with cancer and is increased by surgery, chemotherapy, radiotherapy and disease progression. Low molecular weight heparin (LMWH) is routinely used to treat VTE and some evidence suggests that LMWH may also have an anticancer effect, by reduction in the incidence of metastases. The FRAGMATIC trial will assess the effect of adding dalteparin (FRAGMIN), a type of LMWH, to standard treatment for patients with lung cancer. METHODS/DESIGN: The study design is a randomised multicentre phase III trial comparing standard treatment and standard treatment plus daily LMWH for 24 weeks in patients with lung cancer. Patients eligible for this study must have histopathological or cytological diagnosis of primary bronchial carcinoma (small cell or non-small cell) within 6 weeks of randomisation, be 18 or older, and must be willing and able to self administer 5000 IU dalteparin by daily subcutaneous injection or have it administered to themselves or by a carer for 24 weeks. A total of 2200 patients will be recruited from all over the UK over a 3 year period and followed up for a minimum of 1 year after randomisation. Patients will be randomised to one of the two treatment groups in a 1:1 ratio, standard treatment or standard treatment plus dalteparin. The primary outcome measure of the trial is overall survival. The secondary outcome measures include venous thrombotic event (VTE) free survival, serious adverse events (SAEs), metastasis-free survival, toxicity, quality of life (QoL), levels of breathlessness, anxiety and depression, cost effectiveness and cost utility. TRIAL REGISTRATION: Current Controlled Trials ISRCTN80812769. PMID- 19807918 TI - Two successful pregnancies in a woman with chronic myeloid leukemia exposed to nilotinib during the first trimester of her second pregnancy: case study. AB - The occurrence of chronic myeloid leukemia in pregnancy is rare and its management poses a clinical challenge for physicians treating these patients. We report a 30-year-old woman with chronic myeloid leukemia who became pregnant twice successfully. Philadelphia-positive CML in its chronic phase was diagnosed at 16 weeks of her first gestation. At that time, she received no treatment throughout her pregnancy. At 38 weeks of gestation, a normal infant was delivered by cesarean section. At six weeks postpartum, the patient underwent imatinib mesylate therapy but she could not tolerate the treatment. The treatment was then changed to nilotinib at 400 mg orally b.i.d. Two years later, she became pregnant again while she was on nilotinib 200 mg b.i.d. The unplanned pregnancy was identified during her 7.4 weeks of gestation. Because the patient elected to continue her pregnancy, nilotinib was stopped immediately, and no further treatment was given until delivery. Neither obstetrical complications nor structural malformations in neonates in both pregnancies were observed. Both babies' growth and development have been normal. Although this experience is limited to a single patient, the success of this patient demonstrates that the management of chronic myeloid leukemia in pregnant women may be individualized based on the relative risks and benefits of the patient and fetus. PMID- 19807919 TI - SigWinR; the SigWin-detector updated and ported to R. AB - BACKGROUND: Our SigWin-detector discovers significantly enriched windows of (genomic) elements in any sequence of values (genes or other genomic elements in a DNA sequence) in a fast and reproducible way. However, since it is grid based, only (life) scientists with access to the grid can use this tool. Therefore and on request, we have developed the SigWinR package which makes the SigWin-detector available to a much wider audience. At the same time, we have introduced several improvements to its algorithm as well as its functionality, based on the feedback of SigWin-detector end users. FINDINGS: To allow usage of the SigWin-detector on a desktop computer, we have rewritten it as a package for R: SigWinR. R is a free and widely used multi platform software environment for statistical computing and graphics. The package can be installed and used on all platforms for which R is available. The improvements involve: a visualization of the input-sequence values supporting the interpretation of Ridgeograms; a visualization allowing for an easy interpretation of enriched or depleted regions in the sequence using windows of pre-defined size; an option that allows the analysis of circular sequences, which results in rectangular Ridgeograms; an application to identify regions of co-altered gene expression (ROCAGEs) with a real-life biological use-case; adaptation of the algorithm to allow analysis of non-regularly sampled data using a constant window size in physical space without resampling the data. To achieve this, support for analysis of windows with an even number of elements was added. CONCLUSION: By porting the SigWin-detector as an R package, SigWinR, improving its algorithm and functionality combined with adequate performance, we have made SigWin-detector more useful as well as more easily accessible to scientists without a grid infrastructure. PMID- 19807920 TI - Electroencephalographic correlates of Chronic Fatigue Syndrome. AB - BACKGROUND: Unremitting fatigue and unrefreshing sleep, hallmark traits of Chronic Fatigue Syndrome (CFS), are also pathognomonic of sleep disorders. Yet, no reproducible perturbations of sleep architecture, multiple sleep latency times or Epworth Sleepiness Scores are found to be associated consistently with CFS. This led us to hypothesize that sleep homeostasis, rather than sleep architecture, may be perturbed in CFS. To probe this hypothesis, we measured and compared EEG frequencies associated with restorative sleep between persons with CFS and matched controls, both derived from a population-based sample. METHODS: We evaluated overnight polysomnography (PSG) in 35 CFS and 40 control subjects. PSG records were manually scored and epochs containing artifact removed. Fast Fourier Transformation was utilized to deconstruct individual EEG signals into primary frequency bands of alpha, delta, theta, sigma, and beta frequency domains. The spectral power of each frequency domain for each sleep state was compared between persons with CFS and matched controls. RESULTS: In persons with CFS, delta power was diminished during slow wave sleep, but elevated during both stage 1 and REM. Alpha power was reduced during stage 2, slow wave, and REM sleep. Those with CFS also had significantly lower theta, sigma, and beta spectral power during stage 2, Slow Wave Sleep, and REM. DISCUSSION: Employing quantitative EEG analysis we demonstrate reduced spectral power of cortical delta activity during SWS. We also establish reduced spectral power of cortical alpha activity, with the greatest reduction occurring during REM sleep. Reductions in theta, beta, and sigma spectral power were also apparent. CONCLUSION: Unremitting fatigue and unrefreshing sleep, the waking manifestations of CFS, may be the consequence of impaired sleep homeostasis rather than a primary sleep disorder. PMID- 19807922 TI - Intragenic tandem repeats in Daphnia magna: structure, function and distribution. AB - BACKGROUND: Expressed sequence tag (EST) databases provide a valuable source of genetic data in organisms whose genome sequence information is not yet compiled. We used a published EST database for the waterflea Daphnia magna (Crustacea:Cladocera) to isolate variable number of tandem repeat (VNTR) markers for linkage mapping, Quantitative Trait Loci (QTL), and functional studies. FINDINGS: Seventy-four polymorphic markers were isolated and characterised. Analyses of repeat structure, putative gene function and polymorphism indicated that intragenic tandem repeats are not distributed randomly in the mRNA sequences; instead, dinucleotides are more frequent in non-coding regions, whereas trinucleotides (and longer motifs involving multiple-of-three nucleotide repeats) are preferentially situated in coding regions. We also observed differential distribution of repeat motifs across putative genetic functions. This indicates differential selective constraints and possible functional significance of VNTR polymorphism in at least some genes. CONCLUSION: Databases of VNTR markers situated in genes whose putative function can be inferred from homology searches will be a valuable resource for the genetic study of functional variation and selection. PMID- 19807921 TI - The genome sequence of the protostome Daphnia pulex encodes respective orthologues of a neurotrophin, a Trk and a p75NTR: evolution of neurotrophin signaling components and related proteins in the bilateria. AB - BACKGROUND: Neurotrophins and their Trk and p75NTR receptors play an important role in the nervous system. To date, neurotrophins, Trk and p75NTR have only been found concomitantly in deuterostomes. In protostomes, homologues to either neurotrophin, Trk or p75NTR are reported but their phylogenetic relationship to deuterostome neurotrophin signaling components is unclear. Drosophila has neurotrophin homologues called Spatzles (Spz), some of which were recently renamed neurotrophins, but direct proof that these are deuterostome neurotrophin orthologues is lacking. Trks belong to the receptor tyrosine kinase (RTK) family and among RTKs, Trks and RORs are closest related. Flies lack Trks but have ROR and ROR-related proteins called NRKs playing a neurotrophic role. Mollusks have so far the most similar proteins to Trks (Lymnaea Trk and Aplysia Trkl) but the exact phylogenetic relationship of mollusk Trks to each other and to vertebrate Trks is unknown. p75NTR belongs to the tumor necrosis factor receptor (TNFR) superfamily. The divergence of the TNFR families in vertebrates has been suggested to parallel the emergence of the adaptive immune system. Only one TNFR representative, the Drosophila Wengen, has been found in protostomes. To clarify the evolution of neurotrophin signaling components in bilateria, this work analyzes the genome of the crustacean Daphnia pulex as well as new genetic data from protostomes. RESULTS: The Daphnia genome encodes a neurotrophin, p75NTR and Trk orthologue together with Trkl, ROR, and NRK-RTKs. Drosophila Spz1, 2, 3, 5, 6 orthologues as well as two new groups of Spz proteins (Spz7 and 8) are also found in the Daphnia genome. Searching genbank and the genomes of Capitella, Helobdella and Lottia reveals neurotrophin signaling components in other protostomes. CONCLUSION: It appears that a neurotrophin, Trk and p75NTR existed at the protostome/deuterostome split. In protostomes, a "neurotrophin superfamily" includes Spzs and neurotrophins which respectively form two paralogous families. Trks and Trkl proteins also form closely related paralogous families within the protostomian RTKs, whereby Trkls are absent in deuterostomes. The finding of p75NTR in several protostomes suggests that death domain TNFR superfamily proteins appeared early in evolution. PMID- 19807923 TI - Rethinking the conceptual terrain of AIDS scholarship: lessons from comparing 27 years of AIDS and climate change research. AB - BACKGROUND: While there has recently been significant medical advance in understanding and treating HIV, limitations in understanding the complex social dimensions of HIV/AIDS epidemics continue to restrict a host of prevention and development efforts from community through to international levels. These gaps are rooted as much in limited conceptual development as they are in a lack of empirical research. METHODS: In this conceptual article, the authors compare and contrast the evolution of climate change and AIDS research. They demonstrate how scholarship and response in these two seemingly disparate areas share certain important similarities, such as the "globalization" of discourses and associated masking of uneven vulnerabilities, the tendency toward techno-fixes, and the polarization of debates within these fields. They also examine key divergences, noting in particular that climate change research has tended to be more forward looking and longer-term in focus than AIDS scholarship. CONCLUSION: Suggesting that AIDS scholars can learn from these key parallels and divergences, the paper offers four directions for advancing AIDS research: (1) focusing more on the differentiation of risk and responsibility within and among AIDS epidemics; (2) taking (back) on board social justice approaches; (3) moving beyond polarized debates; and (4) shifting focus from reactive to forward-looking and proactive approaches. PMID- 19807924 TI - Identification of SH3 domain interaction partners of human FasL (CD178) by phage display screening. AB - BACKGROUND: Fas ligand is a cytotoxic effector molecule of T and NK cells which is characterized by an intracellular N-terminal polyproline region that serves as a docking site for SH3 and WW domain proteins. Several previously described Fas ligand-interacting SH3 domain proteins turned out to be crucial for the regulation of storage, expression and function of the death factor. Recent observations, however, indicate that Fas ligand is also subject to posttranslational modifications including shedding and intramembrane proteolysis. This results in the generation of short intracellular fragments that might either be degraded or translocate to the nucleus to influence transcription. So far, protein-protein interactions that specifically regulate the fate of the intracellular fragments have not been identified. RESULTS: In order to further define the SH3 domain interactome of the intracellular region of Fas ligand, we now screened a human SH3 domain phage display library. In addition to known SH3 domains mediating binding to the Fas ligand proline-rich domain, we were able to identify a number of additional SH3 domains that might also associate with FasL. Potential functional implications of the new binding proteins for the death factor's biology are discussed. For Tec kinases and sorting nexins, the observed interactions were verified in cellular systems by pulldown experiments. CONCLUSION: We provide an extended list of putative Fas ligand interaction partners, confirming previously identified interactions, but also introducing several novel SH3 domain proteins that might be important regulators of Fas ligand function. PMID- 19807926 TI - Antioxidant activity of noni juice in heavy smokers. AB - BACKGROUND: Noni (Morinda citrifolia) juice has demonstrated antioxidant activity in vitro and in vivo. To evaluate this activity in humans, noni juice from Tahiti (TNJ) was evaluated in a 30 day, double-blind, and placebo controlled clinical trial with 285 current heavy smokers. Research participants were randomly assigned to three daily treatment groups: 118 mL placebo, 29.5 mL TNJ, and 118 mL TNJ. Plasma superoxide anion radicals (SAR) and lipid hydroperoxide (LOOH) levels were measured pre and post-intervention. RESULTS: After 30 days, mean SAR decreased from 0.26 +/- 0.14 to 0.19 +/- 0.10 micromol/mL in the 29.5 mL dose group (P < 0.01) and from 0.26 +/- 0.22 to 0.18 +/- 0.11 micromol/mL in the 118 mL dose group (P < 0.001). LOOH levels decreased from 0.53 +/- 0.19 to 0.40 +/- 0.10 micromol/mL in the 29.5 mL dose group (P < 0.001) and from 0.55 +/- 0.21 to 0.40 +/- 0.14 micromol/mL in the 118 mL dose group (P < 0.001). No significant reductions in SAR or LOOH levels were observed in the placebo group. CONCLUSION: The results suggest an antioxidant activity from noni juice in humans exposed to tobacco smoke, thereby replicating the results found previous chemical and in vivo tests. PMID- 19807925 TI - Implementation of outpatient schema therapy for borderline personality disorder: study design. AB - BACKGROUND: Schema Therapy (ST) is an integrative psychotherapy based upon a cognitive schema model which aims at identifying and changing dysfunctional schemas and modes through cognitive, experiential and behavioral pathways. It is specifically developed for patients with personality disorders. Its effectiveness and efficiency have been demonstrated in a few randomized controlled trials, but ST has not been evaluated in regular mental healthcare settings. This paper describes the study protocol of a multisite randomized 2-group design, aimed at evaluating the implementation of outpatient schema therapy for patients with borderline personality disorder (BPD) in regular mental healthcare and at determining the added value of therapist telephone availability outside office hours in case of crisis. METHODS/DESIGN: Patient outcome measures will be assessed with a semi-structured interview and self-report measures on BPD, therapeutic alliance, quality of life, costs and general psychopathology at baseline, 6, 12, 18 and 36 months. Intention-to-treat analyses will be executed with survival analysis for dichotomous variables, and one-sample t-tests and ANCOVAs for continuous variables with baseline as covariate and condition as between group factor. All tests will be two-tailed with a significance level of 5%. DISCUSSION: The study will provide an answer to the question whether ST can be effectively implemented and whether phone support by the therapist has an additional value. TRIAL REGISTRATION: The Dutch Cochrane Center, NTR (TC = 1781). PMID- 19807927 TI - The Asia 2 specific signal peptide region and other domains in fusion protein genes characterized Asia 1 and Asia 2 canine distemper viruses. AB - BACKGROUND: Although the presence of Asia 2 group of canine distemper virus (CDV) was known by the sequencing and phylogenetic analysis of hemagglutinin (H) gene, the fusion (F) protein gene sequence of Asia 2 group had not been identified. So, the sequence analysis of F gene was carried out to elucidate the genotypic varaitons among Asian isolates. RESULTS: The phylogenetic analysis of F and H gene sequences from fourteen CDV isolates obtained from diseased dogs in Japan and Thailand indicated that the F genes had a new initiation codon and extra 27 nucleotides upstream of the usual open reading frame (ORF) and the F proteins had extra 9 amino acids at the N-terminal position only in Asia 2 isolates. On the contrary, the Asia 1 isolates had three extra putative N-glycosylation sites (two sites in the signal peptide region and one site in the F1 region) except for two strains of Th12 and Ac96I (two sites in signal peptide region) adding to four putative N-glycosylation sites that were conserved among all Asian isolates and Onderstepoort strain. In addition to this difference in N-glycosylation sites, the signal peptide region had a great diversity between Asia 1 and Asia 2 isolates. Also, characteristic amino acids were detected for some strains. CONCLUSION: Asia 2 isolates were distinguished from other CDV lineages by the extra 27 nucleotide sequence. The signal peptide region of F gene gives a remarkable differentiation between Asia 1 and Asia 2 isolates. Strains Th12 and Ac96I were differentiated from other Asia 1 strains by the F protein glycosylation sites. PMID- 19807928 TI - Paramedic assessment of pain in the cognitively impaired adult patient. AB - BACKGROUND: Paramedics are often a first point of contact for people experiencing pain in the community. Wherever possible the patient's self report of pain should be sought to guide the assessment and management of this complaint. Communication difficulty or disability such as cognitive impairment associated with dementia may limit the patient's ability to report their pain experience, and this has the potential to affect the quality of care. The primary objective of this study was to systematically locate evidence relating to the use of pain assessment tools that have been validated for use with cognitively impaired adults and to identify those that have been recommended for use by paramedics. METHODS: A systematic search of health databases for evidence relating to the use of pain assessment tools that have been validated for use with cognitively impaired adults was undertaken using specific search criteria. An extended search included position statements and clinical practice guidelines developed by health agencies to identify evidence-based recommendations regarding pain assessment in older adults. RESULTS: Two systematic reviews met study inclusion criteria. Weaknesses in tools evaluated by these studies limited their application in assessing pain in the population of interest. Only one tool was designed to assess pain in acute care settings. No tools were located that are designed for paramedic use. CONCLUSION: The reviews of pain assessment tools found that the majority were developed to assess chronic pain in aged care, hospital or hospice settings. An analysis of the characteristics of these pain assessment tools identified attributes that may limit their use in paramedic practice. One tool--the Abbey Pain Scale--may have application in paramedic assessment of pain, but clinical evaluation is required to validate this tool in the paramedic practice setting. Further research is recommended to evaluate the Abbey Pain Scale and to evaluate the effectiveness of paramedic pain management practice in older adults to ensure that the care of all patients is unaffected by age or disability. PMID- 19807929 TI - The influence of P-glycoprotein expression and its inhibitors on the distribution of doxorubicin in breast tumors. AB - BACKGROUND: Anti-cancer drugs access solid tumors via blood vessels, and must penetrate tumor tissue to reach all cancer cells. Previous studies have demonstrated steep gradients of decreasing doxorubicin fluorescence with increasing distance from blood vessels, such that many tumor cells are not exposed to drug. Studies using multilayered cell cultures show that increased P glycoprotein (PgP) is associated with better penetration of doxorubicin, while PgP inhibitors decrease drug penetration in tumor tissue. Here we evaluate the effect of PgP expression on doxorubicin distribution in vivo. METHODS: Mice bearing tumor sublines with either high or low expression of PgP were treated with doxorubicin, with or without pre-treatment with the PgP inhibitors verapamil or PSC 833. The distribution of doxorubicin in relation to tumor blood vessels was quantified using immunofluorescence. RESULTS: Our results indicate greater uptake of doxorubicin by cells near blood vessels in wild type as compared to PgP overexpressing tumors, and pre-treatment with verapamil or PSC 833 increased uptake in PgP-overexpressing tumors. However, there were steeper gradients of decreasing doxorubicin fluorescence in wild-type tumors compared to PgP overexpressing tumors, and treatment of PgP overexpressing tumors with PgP inhibitors led to steeper gradients and greater heterogeneity in the distribution of doxorubicin. CONCLUSION: PgP inhibitors increase uptake of doxorubicin in cells close to blood vessels, have little effect on drug uptake into cells at intermediate distances, and might have a paradoxical effect to decrease doxorubicin uptake into distal cells. This effect probably contributes to the limited success of PgP inhibitors in clinical trials. PMID- 19807930 TI - A cost-based equity weight for use in the economic evaluation of primary health care interventions: case study of the Australian Indigenous population. AB - BACKGROUND: Efficiency and equity are both important policy objectives in resource allocation. The discipline of health economics has traditionally focused on maximising efficiency, however addressing inequities in health also requires consideration. Methods to incorporate equity within economic evaluation techniques range from qualitative judgements to quantitative outcomes-based equity weights. Yet, due to definitional uncertainties and other inherent limitations, no method has been universally adopted to date. This paper proposes an alternative cost-based equity weight for use in the economic evaluation of interventions delivered from primary health care services. METHODS: Equity is defined in terms of 'access' to health services, with the vertical equity objective to achieve 'equitable access for unequal need'. Using the Australian Indigenous population as an illustrative case study, the magnitude of the equity weight is constructed using the ratio of the costs of providing specific interventions via Indigenous primary health care services compared with the costs of the same interventions delivered via mainstream services. Applying this weight to the costs of subsequent interventions deflates the costs of provision via Indigenous health services, and thus makes comparisons with mainstream more equitable when applied during economic evaluation. RESULTS: Based on achieving 'equitable access', existing measures of health inequity are suitable for establishing 'need', however the magnitude of health inequity is not necessarily proportional to the magnitude of resources required to redress it. Rather, equitable access may be better measured using appropriate methods of health service delivery for the target group. 'Equity of access' also suggests a focus on the processes of providing equitable health care rather than on outcomes, and therefore supports application of equity weights to the cost side rather than the outcomes side of the economic equation. CONCLUSION: Cost-based weights have the potential to provide a pragmatic method of equity weight construction which is both understandable to policy makers and sensitive to the needs of target groups. It could improve the evidence base for resource allocation decisions, and be generalised to other disadvantaged groups who share similar concepts of equity. Development of this decision-making tool represents a potentially important avenue for further health economics research. PMID- 19807932 TI - A practical and transferable new protocol for treadmill testing of children and adults? PMID- 19807931 TI - The absorption and uptake of recombinant human follicle-stimulating hormone through vaginal subcutaneous injections--a pharmacokinetic study. AB - BACKGROUND: Follicle stimulating hormone (FSH) has been routinely used for ovulation induction. Because of rapid clearance of the hormone, FSH is commonly administered by daily intramuscular or subcutaneous injections in in-vitro fertilization (IVF). To reduce the number of visits to the clinic, an intermittent vaginal injection of rhFSH every 3 days employing the concepts of mesotherapy and uterine first-pass effect was invented and has successfully been applied in women receiving IVF treatment. This study was designed to monitor the pharmacokinetic pattern of rhFSH administered vaginally. METHODS: Twelve healthy women with regular ovulatory cycles were recruited. All volunteers received gonadotrophin-releasing hormone agonist to suppress pituitary function and were assigned to receive single dose recombinant human FSH (rhFSH, Puregon 300) either using conventional abdominal subcutaneous injection or vaginal subcutaneous injection in a randomized cross-over study. Serum samples were collected at pre- scheduled time intervals after injections of rhFSH to determine immunoreactive FSH levels. Pharmacokinetic parameters characterizing rate [maximal plasma concentrations (Cmax) and time of maximal plasma concentrations (tmax)] and extent [area under the plasma concentration-time curve (AUC) and clearance] of absorption of rhFSH were compared. RESULTS: Vaginal injection of rhFSH was well tolerated and no drug-related adverse reaction was noted. Our analysis revealed that tmax was significantly earlier (mean 6.67 versus 13.33 hours) and Cmax was significantly higher (mean 17.77 versus 13.96 IU/L) in vaginal versus abdominal injections. The AUC(0-infinity) was 1640 versus 1134 IU hour/L in vaginal and abdominal injections, respectively. Smaller plasma elimination rate constant (0.011 versus 0.016 hour-1), longer mean residence time (106.58 versus 70.47 hours), and slower total body clearance (292.2 versus 400.1 mL/hour) were also found in vaginal injection. CONCLUSION: The vaginal injection mode elicited a rapid and highly extended absorption of rhFSH injected compared to conventional abdominal injection. These data indicate that the rate and extent of FSH absorption from the injection site can vary depending on the route of the FSH administration. PMID- 19807933 TI - Fruit and vegetable intake in Austrian adults: intake frequency, serving sizes, reasons for and barriers to consumption, and potential for increasing consumption. AB - OBJECTIVE: To assess the intake frequency of fruit and vegetables, serving sizes, reasons for and barriers to consumption, and the potential for increasing fruit and vegetable intake. DESIGN: A nationwide postal questionnaire survey was conducted in 2006 over all four seasons. The participants were stratified according to occupation and sex. The response rate for 5130 questionnaires sent out was 52.7 %. SETTING: Austria. SUBJECTS: Austrian adults, aged 19-64 years. RESULTS: Daily fruit consumption was reported by 57.1 % of the participants and daily vegetable consumption by 36.2 %. On average, 2.1 (SD 1.9) servings (250 (SD 225) g) of fruit and 1.7 (SD 1.3) servings (198 (SD 159) g) of vegetables were consumed daily. Women ate fruit and vegetables both more frequently and in greater quantities than men. Both intake frequency and the number of fruit and vegetable servings were largely independent of seasonal fluctuations. The primary reason for the consumption of both fruit and vegetables was taste. The greatest barrier to higher intake was the perception that current individual consumption was already sufficient. Price did not constitute a relevant barrier in Austria. At present, the potential for increasing fruit and vegetable intake can be estimated at two servings. CONCLUSIONS: Austrian adults still consume less fruit and vegetables than recommended. Strategies to increase intake should pay more attention to the taste instead of the various health aspects. PMID- 19807934 TI - Dietary carbohydrates and change in physical performance of elderly Europeans: Survey in Europe on Nutrition and the Elderly, a Concerted Action (SENECA) 1993 and 1999. AB - OBJECTIVE: To study dietary glycaemic index (GI) and glycaemic load (GL) in association with physical performance in elderly Europeans. DESIGN: Cross sectional and prospective study. Physical performance was measured using the Physical Performance Test (PPT) score on a scale from 0 to 27, where high scores indicate a better physical performance. Habitual diets were measured using diet history interviews and dietary GI and GL were estimated from table values. SETTING: Eight towns/centres from the Survey in Europe on Nutrition and the Elderly, a Concerted Action (SENECA) in 1993 and 1999. SUBJECTS: Seven hundred and sixty-five men and women, 75-80 years old, were examined in 1993; of these, 357 (47 %) were followed up in 1999, at age 80-85 years. RESULTS: At baseline, both dietary GI and GL were significantly inversely associated with PPT scores (P = 0.03 and P = 0.05, respectively). When adjusted for age, BMI, physical activity, self-perceived health, chronic diseases and town/centre, the strength of the associations was attenuated and became non-significant (GI, P = 0.08; GL, P = 0.92). Dietary GI/GL were not associated with PPT scores 6 years later. CONCLUSIONS: Among elderly Europeans, a high glycaemic diet was associated with a low physical performance at baseline but not 6 years later. Cross-sectional associations may in part be caused by variations in age, BMI, physical activity, self-perceived health, chronic diseases and geographic location. PMID- 19807935 TI - Malnutrition in free-living elderly in rural south India: prevalence and risk factors. AB - OBJECTIVE: To estimate the prevalence of malnutrition among free-living elderly in a rural population of south India. DESIGN: Cross-sectional study. Nutritional status was assessed using the Mini Nutritional Assessment (MNA) questionnaire, which is an eighteen-item nutritional screening instrument used in the elderly. SETTING: Kaniyambadi block, a rural development block in the state of Tamil Nadu, south India. SUBJECTS: Community-dwelling elderly (aged 60 years and above). RESULTS: As evaluated by the MNA, 14 % of the 227 subjects were malnourished and 49 % were at risk of malnourishment. No significant difference was found between men and women. The majority of the elderly were living with their children, had no income and consumed three meals per day. Older age (P < 0.001), decreased food intake (P < 0.001) and consuming fewer meals (P < 0.001) were independently associated with lower MNA scores. CONCLUSIONS: More than 60 % of the subjects had low MNA scores (<23.5) indicating that deficient protein-energy intake is common among rural elderly of south India and requires more attention. PMID- 19807936 TI - Overweight and weight dissatisfaction related to socio-economic position, integration and dietary indicators among south Asian immigrants in Oslo. AB - OBJECTIVE: To investigate how socio-economic position, demographic factors, degree of integration and dietary indicators are related to BMI/waist:hip ratio (WHR) and to weight dissatisfaction and slimming among South Asians in Oslo, Norway. DESIGN: Cross-sectional study consisting of a health check including anthropometric measures and two self-administered questionnaires. SETTING: Oslo, Norway. SUBJECTS: Pakistanis and Sri Lankans (n 629), aged 30-60 years, residing in Oslo. RESULTS: BMI was positively associated with female gender (P = 0.004) and Pakistani origin (P < 0.001), and inversely associated with years of education (P = 0.011) and eating more hot meals (P = 0.016). WHR was positively associated with male gender (P < 0.001), age (P < 0.001) and a dietary pattern with high-fat foods (P = 0.005), and inversely associated with degree of integration (measured by a composite index, independent of duration of residence; P = 0.017). One-third of those with normal weight and most of those obese were dissatisfied with their weight. Among these, about 40 % had attempted to slim during the past year. Dissatisfaction with weight was positively associated with education in women (P = 0.006) and with integration in men (P = 0.026), and inversely associated with physical activity (P = 0.044) in men. Women who had made slimming attempts had breakfast and other meals less frequently than others (P < 0.05). CONCLUSIONS: Weight dissatisfaction exists among South Asian immigrants. More research is needed regarding bodily dissatisfaction and the relationship between perception of weight and weight-change attempts among immigrants in Norway, in order to prevent and treat both obesity and eating disorders. PMID- 19807937 TI - Reliability and relative validity of an FFQ for nutrients in the Tehran lipid and glucose study. AB - OBJECTIVE: To describe the relative validity and reliability of the FFQ used for assessing nutrient intakes of participants in the Tehran Lipid and Glucose Study (TLGS). DESIGN: A total of 132 subjects (sixty-one males and seventy-one females) were included in the study. Dietary data were collected monthly by means of twelve 24 h dietary recalls (24hDR). Subjects completed two, 168-item semi quantitative FFQ. Blood and urine samples were taken every season for measurement of plasma biomarkers and urinary N and K. RESULTS: Mean age and BMI of the participants were 35.5 (sd 16.8) years and 25.5 (sd 5.2) kg/m2, respectively. The mean energy-adjusted and deattenuated correlation coefficients for overall nutrient intake between the 24hDR and FFQ2 were 0.44 and 0.37 in 35-year-olds, respectively, and for individual nutrients ranged from 0.24 to 0.71 in men (mean r = 0.53) and from 0.11 to 0.60 in women (mean r = 0.39). The mean energy-adjusted reliability coefficients varied from 0.48 in 35-year-olds, and ranged from 0.41 to 0.79 in men (mean r = 0.59) and from 0.39 to 0.74 in women (mean r = 0.60). The FFQ2 and 24hDR produced exact agreement rates ranging between 39.6 % and 68.3 % in men and between 39.6 % and 54.1 % in women. The ranges of questionnaire validity coefficients, with the sample correlation between the questionnaires and biochemical marker as the lower limit and the estimate obtained by the method of triads as the upper limit, were 0.21-0.56 (protein) and 0.37-0.61 (K). CONCLUSIONS: The FFQ developed for the TLGS has reasonable relative validity and reliability for nutrient intakes in Tehranian adults. PMID- 19807938 TI - Association between breast-feeding and anthropometry and CVD risk factor status in adolescence and young adulthood: the Young Hearts Project, Northern Ireland. AB - OBJECTIVE: To examine the association between breast-feeding and blood pressure, anthropometry and plasma lipid profile in both adolescence and young adulthood. DESIGN: Longitudinal study of biological and behavioural risk factors for CVD. SETTING: The Young Hearts Project, Northern Ireland. SUBJECTS: Schoolchildren aged 12 years and 15 years who participated in a cross-sectional study of lifestyle and health, and who were followed up as young adults aged 20-25 years. RESULTS: There was no significant difference in height, weight, BMI, skinfold thickness measurements, blood pressure or plasma lipid profile in adolescents who had been breast-fed compared with those who had not been breast-fed. However, by the time these adolescents had reached adulthood, those who had been breast-fed were significantly taller than those who had not been breast-fed (standing height, P = 0.013; leg length, P = 0.035). Specifically, the breast-fed group was on average taller by 1.7 cm (95 % CI 0.4, 3.0 cm) and had longer legs by 1.0 cm (95 % CI 0.1, 1.9 cm). There was no significant difference in other anthropometric measures, blood pressure or plasma lipid profile in adults who had been breast-fed compared with those who had not been breast-fed. CONCLUSIONS: Compared with those who had not been breast-fed, individuals who had been breast fed were taller in adulthood. Given the known association of increased adult height with improved life expectancy, the results from the present study support a beneficial effect of breast-feeding. PMID- 19807939 TI - An empirical study of Taiwan's food security index. AB - OBJECTIVE: The FAO has developed an approach for estimating the prevalence of undernourishment. Based on the FAO method Taiwan has a prevalence of undernourishment of 3.98%, which is higher than that of some developing countries. As this is not a true reflection of the status of undernourishment in our nation, the purpose of the present study was to modify the FAO methodology for Taiwan. DESIGN: Two factors were considered in the modified version. As the minimum dietary energy requirement was the main factor contributing to the inflated prevalence in Taiwan, we adjusted for a lighter physical activity level, based on the average BMI of the Taiwanese population, and calculated a new minimum dietary energy requirement. We then fitted a second-order polynomial regression model for prediction of per capita dietary energy supply. RESULTS: The adjusted minimum dietary energy requirement was reduced to 7648 kJ/d or 7765 kJ/d compared with the original value of 8054 kJ/d. This resulted in a decrease of the prevalence of undernourishment in Taiwan to 2.5% or 3.0%, which is much closer to that of other countries with the same level of economic development. The second order polynomial regression model efficiently reduced the variation in dietary energy consumption and resulted in an undernourishment prevalence of less than 2.5%. CONCLUSIONS: This new adapted method is more appropriate for Taiwan. It is recommended that each country evaluates the appropriateness of the FAO approach for its population. PMID- 19807940 TI - Cognitive and affective sequelae of primary hyperparathyroidism and early response to parathyroidectomy. AB - Cognitive and affective complaints are common in patients with primary hyperparathyroidism (PHPT), but few studies have used psychometric testing to document these symptoms and their response to parathyroidectomy. The current study sought to clarify the nature of cognitive and affective impairments in PHPT and changes postparathyroidectomy. One hundred eleven patients with PHPT underwent neuropsychological evaluation prior to parathyroidectomy with 68 returning for an early postsurgical evaluation. Changes in cognition were assessed using practice effect corrected reliable change indices. Biochemical and anesthesia variables were compared between groups who improved and declined. In a subset of patients, assessment revealed a significant pattern of cognitive slowing, reductions in psychomotor speed, memory impairment, and depression prior to parathyroidectomy. Postsurgical evaluations revealed a trend for improvements on timed tests and depression but a decline in memory. Older patients responded less well to surgical intervention, as did patients who experienced more dramatic changes in biochemical status following surgery. Cognitive changes early postparathyroidectomy are characterized by improved information processing speed and decline in verbal memory, with younger patients more likely to recover during this acute phase. The need for longer-term follow-up studies and increasing utilization of neuropsychological assessments in this population are discussed. PMID- 19807941 TI - Fate determination of adult human glial progenitor cells. AB - Glial progenitor cells (GPCs) comprise the most abundant population of progenitor cells in the adult human brain. They are responsible for central nervous system (CNS) remyelination, and likely contribute to the astrogliotic response to brain injury and degeneration as well. Adult human GPCs are biased to differentiate as oligodendrocytes and elaborate new myelin, and yet they retain multilineage plasticity, and can give rise to neurons as well as astrocytes and oligodendrocytes once removed from the adult parenchymal environment. GPCs retain strong mechanisms for cell-autonomous self-renewal, and yet both their phenotype and fate may be dictated by their microenvironment. Using the transcriptional profiles of acutely isolated GPCs, we have begun to understand the operative ligand-receptor interactions involved in these processes, and have identified several key signaling pathways by which adult human GPCs may be reliably instructed to either oligodendrocytic or astrocytic fate. In addition, we have noted significant differences between the expressed genes and dominant signaling pathways of fetal and adult human GPCs, as well as between rodent and human GPCs. The latter data in particular call into question therapeutic strategies predicated solely upon data obtained using rodents, while perhaps highlighting the extent to which evolution has been attended by the phylogenetic modification of glial phenotype and function. PMID- 19807942 TI - What will the others think? In-group norms as a mediator of the effects of intergroup contact. AB - The influence of social norms in the context of intergroup relations has long been recognized by social psychologists, yet research on intergroup contact and social norms have usually remained disconnected. We explored the influence of direct and indirect friendship on attitudes towards ethnic minorities in Norway, and in particular the role of in-group norms about the social approval of intergroup contact as a mechanism that distinguishes direct from indirect contact. Using a sample of school students from 89 classrooms (N=823), we tested this hypothesis with both one level and multi-level structural equation modelling (ML-SEM), where the amount of contact of other classroom members was considered as a form of indirect contact. The results suggest that the intergroup contact of other in-group members (in-group friends or classmates) affects attitudes towards the out-group by changing the perception of in-group norms and by reducing intergroup anxiety. In contrast, direct contact (or contact at the individual level in the case of ML-SEM), improved attitudes only by reducing intergroup anxiety, and did not affect the perception of in-group norms. PMID- 19807943 TI - Individual differences and reasoning: a study on personality traits. AB - Personality can play a crucial role in how people reason and decide. Identifying individual differences related to how we actively gather information and use evidence could lead to a better comprehension and predictability of human reasoning. Recent findings have shown that some personality traits are related to similar decision-making patterns showed by people with mental disorders. We performed research with the aim to investigate delusion-proneness, obsessive-like personality, anxiety (trait and state), and reasoning styles in individuals from the general population. We introduced personality trait and state anxiety scores in a regression model to explore specific associations with: (1) amount of data gathered prior to making a decision; and (2) the use of confirmatory and disconfirmatory evidence. Results showed that all our independent variables were positively or negatively associated with the amount of data collected in order to make simple probabilistic decisions. Anxiety and obsessiveness were the only predictors of the weight attributed to evidence in favour or against a hypothesis. Findings were discussed in relation to theoretical assumptions, predictions, and clinical implications. Personality traits can predict peculiar ways to reason and decide that, in turn, could be involved to some extent in the formation and/or maintenance of psychological disorders. PMID- 19807944 TI - Teacher organizational citizenship behaviours and job efficacy: Implications for student quality of school life. AB - The present study investigated the impact of teachers' organizational citizenship behaviours (OCBs) on student quality of school life (SQSL) via the indirect effect of job efficacy. A measure of teacher OCBs was developed, tapping one dimension of individual-focused OCB (OCBI - student-directed behaviour) and two dimensions of organization-focused OCB (OCBO - civic virtue and professional development). In line with previous research suggesting that OCBs may enhance job efficacy, as well as studies demonstrating the positive effects of teacher efficacy on student outcomes, we expected an indirect relationship between teachers OCBs and SQSL via teachers' job efficacy. Hypotheses were tested in a multi-level design in which 170 teachers and their students (N=3,057) completed questionnaires. A significant proportion of variance in SQSL was attributable to classroom factors. Analyses revealed that the civic virtue and professional development behaviours of teachers were positively related to their job efficacy. The job efficacy of teachers also had a positive impact on all five indicators of SQSL. In regards to professional development, job efficacy acted as an indirect variable in the prediction of four student outcomes (i.e., general satisfaction, student-teacher relations, achievement, and opportunity) and fully mediated the direct negative effect on psychological distress. PMID- 19807945 TI - Reactive aggression in psychopathy and the role of frustration: susceptibility, experience, and control. AB - This article is a commentary on 'Psychopathy, frustration, and reactive aggression: The role of ventromedial prefrontal cortex' (Blair, 2010). PMID- 19807946 TI - Comparing the regression slopes of independent groups. AB - In this study, we explore the effects of non-normality and heteroscedasticity when testing the hypothesis that regression lines associated with two independent groups have the same slopes. Our results indicate that some recently proposed methods that allow heteroscedasticity and perform well in extant simulation studies do not perform well for the situation at hand. Two of the methods studied here are recommended for general use. PMID- 19807947 TI - Flagging fasting plasma glucose specimens: time to routinely label the context in which pathology specimens are recorded. PMID- 19807948 TI - Hospital data may be more accurate than census data in estimating the ethnic composition of general practice populations. AB - BACKGROUND: Equity of service provision by age, ethnicity and sex is a key aim of Government policy in the UK. The prevalence, natural history and management of common chronic conditions, such as diabetes and hypertension, vary between ethnic groups. Developing and monitoring responsive local services requires accurate measures of ethnicity and language needs. Hence establishing the ethnic composition of GP populations is important. OBJECTIVE: To compare three methods of estimating the ethnic composition of GP registered populations in three east London primary care trusts (PCTs). DESIGN: Self-reported ethnicity, routinely collected at practice level (and considered the 'gold standard'), was compared with two indirect methods of attributing ethnicity. The indirect method currently used in the UK assigns ethnicity to GP populations based on geographical postcode attribution from the national census. A proposed alternative indirect method uses the ethnic breakdown of hospital admission data from practice lists to attribute ethnicity to the whole practice population. Comparisons were made between practice self-report recording and these two indirect methods. Bland-Altman plots were used to assess the agreement between methods of measurement. RESULTS: Data from 103 practices, covering 70% of the GP registered population, was used. The hospital admission method showed better agreement with practice self-report data than the census attributed method. For white populations Bland-Altman plots showed a mean difference of 1.4% (95% CI-14.9 to 17.7) between hospital admission and practice data, and a mean difference of 12.5% (95% CI-6.2 to 31.1) between census attributed and practice data. Differences were also found for south Asian and black populations. CONCLUSION: Practice ethnicity measured using hospital attendance data is in closer agreement with practice recording of self-reported ethnicity than the census attribution method. Census attribution may provide misleading information on the ethnic composition of practice populations. We recommend that healthcare commissioners change to this method of measurement when practice self-report data is not available. PMID- 19807949 TI - The cognitive impact of research synopses on physicians: a prospective observational analysis of evidence-based summaries sent by email. AB - BACKGROUND: Effective information transfer in primary care is becoming more difficult as the volume of medical information expands. Emailed research synopses are expected to raise awareness and thereby permit more effective information retrieval. OBJECTIVE: To identify key factors that influence physicians' self reported cognitive impact of emailed research synopses. METHOD: In this prospective observational study, research synopses sent by email between 8 September 2006 and 30 May 2007 were analysed. Seven characteristics of synopses (number of characters, research design, study setting, number of types of patient populations studied, number of comparisons, number of outcomes, and number of results) were analysed. Each synopsis was classified as either positive or negative based on physician-reported impacts. Logistic regression analysis was used to evaluate the association between a negative impact and the synopsis' characteristics. RESULTS: A total of 1960 Canadian physicians submitted 159,442 ratings on 193 synopses. Each synopsis was assessed on average by 826.1 physicians. On average there were 28.3 negative ratings per research synopsis, 146.3 neutral, and 656.2 positive. Out of the seven characteristics analysed, only the number of comparisons (odds ratio (OR) = 0.47, 95% confidence interval (CI) = 0.23-0.93) and the number of results (OR = 0.64, 95% CI = 0.44-0.93) had a statistically significant influence on physician ratings. An increase in the number of comparisons (P = 0.03) or the number of results (P = 0.02) decreased the likelihood of a negative impact. CONCLUSIONS: Characteristics of the synopses appear to influence cognitive impact, and there might be lexical patterns specific to these factors. Further research is recommended in order to understand the mechanism for the influence of these characteristics. PMID- 19807950 TI - Leveraging time and learning style, iPod vs. realtime attendance at a series of medicine residents conferences: a randomised controlled trial. AB - OBJECTIVE: To determine whether participation in educational conferences utilising iPod technology enhances both medical knowledge and accessibility to educational content among medical residents in training. DESIGN/MEASUREMENTS: In May 2007, the authors led a randomised controlled study involving 30 internal medicine residents who volunteered either to attend five midday educational conferences or to use an iPod audio/video recording of the same conferences, each followed by a five-question competency quiz. Primary outcomes included quantitative assessment of knowledge acquisition and qualitative assessment of resident perception of ease of use. Secondary outcomes included resident perception of self-directed learning. RESULTS: At baseline, residents reported attendance at 50% of educational conferences. Of iPod participants, 46.7% previously used an iPod. During the study, 46-60% of conference attendees were paged out of each conference, of whom between 6 and 33% missed more than half of the conference. The quiz completion rate was 93%. Key findings were: 1) similar quiz scores were achieved by conference attendees, mean 60.7% (95% CI; 53.0 68.3%), compared to the iPod user group, mean 67.6% (95% CI; 61%-74.1%), and 2) the majority (10/15, 66.6%) of conference attendees stated they would probably benefit from the option to refer back to conferences for content review and educational purposes. CONCLUSIONS: Residency training programmes can optimise time management strategies with the integration of innovative learning resources into educational curricula. This study suggests that iPod capture of conferences is a reasonable resource to help meet the educational goals of residents and residency programs. PMID- 19807951 TI - Clinical informatics to improve quality of care: a population-based system for patients with diabetes mellitus. AB - BACKGROUND: The prevalence of diabetes mellitus is increasing in the USA. However, control of intermediate outcome measures remains substandard. Recently, significant emphasis has been placed on the value of electronic medical records and informatics systems to improve the delivery of health care. OBJECTIVE: To determine whether a clinical informatics system improves care of patients with diabetes mellitus. METHODS: In this quality improvement pilot initiative, we identified 48 patients with diabetes mellitus who were due for their annual haemoglobin A1c (HbA1c), low-density lipoprotein (LDL) and microalbumin tests. Through our newly developed clinical informatics initiative, patients were reminded to schedule tests and a physician appointment. Seventy-five patients without reminders served as controls. RESULTS: A significant improvement in LDL control was achieved in the intervention group (35.4% vs 13.3%; P=0.004). The intervention group had a greater percentage of patients who underwent the three tests, and members of this group also showed greater control of haemoglobin A1c, but these differences were not statistically significant. CONCLUSIONS: A clinical informatics system, used to deliver proactive, co-ordinated care to a population of patients with diabetes mellitus, can improve process and also quality outcome measures. Larger studies are needed to confirm these early findings. PMID- 19807952 TI - Record media used by primary care providers in medically underserved regions of upstate New York was not pivotal to clinical result in the Informatics for Diabetes Education and Telemedicine (IDEATel) project. AB - PURPOSE: To examine integration of electronic medical records (EMRs) by primary care providers (PCPs) in a diabetes telemedicine project (IDEATel) in medically underserved rural areas and assess if access to digital records is associated with diabetes intermediate outcomes. METHOD: PCPs (n=61) with patients in IDEATel participated in structured interviews to determine current (2006 to 2007) and projected (2007 to 2008) use of paper and/or electronic medical data. T-tests examined group differences. RESULTS: 28% (17/61) of PCPs had comprehensive EMRs, but most electronic data were non-interoperative between offices; 6% of PCPs solely used paper; 92% of PCPs used mixed paper/electronic records. Half of 61 PCPs anticipated no migration within one year to an electronic record for common patient data, while one third anticipated that function would become greatly more electronic. Among 31 PCPs interviewed in depth in person, 70% (7/10) in private practice and 69% (9/13) in networks anticipated greater electronic media migration through system change, whereas 100% of responding academic PCPs (n=6) expected only system modifications. PCPs were most interested in data exchange for chronic disease management (94%), regional benchmarking (84%) and quality improvement (87%). Patient personal electronic health records were rarely mentioned. IDEATel patients of PCPs with or without access to comprehensive EMRs achieved similar haemoglobin A1c, blood pressure, LDL-cholesterol, and body mass index, but the small number invokes cautious interpretation. CONCLUSIONS: Our findings suggest an effective and complementary element of national health information technology (HIT) strategy, telemedicine, can be implemented by PCPs with success despite the lack of a concurrent EMR for efficient data exchange. PMID- 19807953 TI - Variation in the recording of diabetes diagnostic data in primary care computer systems: implications for the quality of care. AB - BACKGROUND: Diabetes mellitus (DM) is a serious, chronic condition affecting 2.3 million people in the UK and consuming over 5% of the total National Health Service (NHS) budget. The World Health Organization (WHO) has produced a classification of diabetes which should help ensure consistent diagnosis and management of cases. However, recent quality based targets for diabetes in the UK only allow for people with Type 1 or Type 2 diabetes to be included in the disease register. OBJECTIVE: To analyse the codes offered when recording a diagnosis of diabetes in an electronic patient record (EPR) system and to assess what proportion of existing codes would map to known diagnostic categories. METHOD: Code-sets (4-byte, 5-byte, CTv3 and SNOMED-CT) were sourced using the NHS Triset Browser and the SNOMED-CT website. We analysed the variation in child codes listed under 'diabetes mellitus'. Picking lists were generated across four general practices, using eight search terms. We examined list length and the types of codes offered. An attempt was also made to map current codes to the WHO classification of diabetes, defining each as having a 'direct mapping', a 'possible mapping', or 'no clear mapping'. RESULTS: SNOMED-CT provided a more concise list of codes (115) than the more widely used 5-byte code-set (177). There was considerable variation in the codes offered in picking lists, with variation occurring between systems, rather than between individual GP practices. In considering the potential for mapping between current code-sets and the WHO classification, there was a general downward trend in the number that had 'no clear mapping' (5-byte Read codes--46.3%, SNOMED-CT--19.1%). CONCLUSION: There is considerable variation in the different diabetic coding hierarchies and in the choices offered at the point of coding in an EPR system. This is likely to lead to inconsistent data recording. Migrating GP computer systems to SNOMED-CT or to another more limited coding system which would map to international disease classifications would enable primary care EPR systems to better support improved standards of care. PMID- 19807954 TI - Using surrogate markers in primary electronic patient record systems to confirm or refute the diagnosis of diabetes. AB - BACKGROUND: UK primary care records are computerised and these records are used for both research and quality improvement. However, there is disparity in the prevalence of diabetes found in epidemiological studies compared with that reported through the UK's national quality improvement scheme. OBJECTIVE: To investigate how non-diagnostic computer data could be used to identify, confirm or refute prevalent cases of people with diabetes. METHOD: We carried out a literature review to identify the most accurate non-diagnostic markers. For each type of diabetes we focused on four broad areas; demographic details, biochemical markers, clinical features and therapeutic strategies. Sample markers were tested by calculating their positive predictive value (PPV) and sensitivity (Sn) and their ability to differentiate between types of diabetes. RESULTS: Biochemical markers were useful in identifying cases of diabetes but not in differentiating between types of diabetes as the same plasma glucose criterion is used to diagnose Type 1, Type 2, and 'other' types of diabetes; the lack of a 'fasting' qualifier blunts the use of this marker. Auto-immune markers were the most accurate in identifying Type 1 diabetes but are not recorded frequently in primary care. Clinical features of diabetes and therapeutic strategies were of some use--however, without time sequence data are difficult to interpret. Raised plasma glucose (PG), and glycated haemoglobin (HbA1c), had useful PPV but low Sn. When PG was more than 7.0 and less than 11.1 mmol/l, PPV equalled 77.8% and Sn 48%; and when PG was 11.1, PPV equalled 92% and Sn 17%. For an HbA1c of more than 6.5%, PPV was 89% and Sn 73.3%, and for an HbA1c of more than 8, PPV was 92% and Sn 26%. A person with a combination of aged under 30 years and prescribed insulin has an 84% PPV of Type 1 diabetes; if they also have a BMI <30 kg/m2 the PPV increases to 88%. A person age over 45 years and with a BMI >30 kg/m2 has a 5.3% PPV of Type 2 diabetes; if they are also hypertensive the PPV is 30%; Asian ethnicity increases PPV to 44%. CONCLUSION: Non-diagnostic data has the potential to confirm or refute the diagnosis of diabetes and identify its type. PMID- 19807955 TI - Illogical placing of codes within a clinical classification. PMID- 19807956 TI - Is it time to take primary care seriously in redressing health inequalities? PMID- 19807957 TI - Quality of care in two health systems: can the United Kingdom and Australia learn from each other? PMID- 19807958 TI - Improving care in depression: qualitative study investigating the effects of using a mental health questionnaire. AB - BACKGROUND: Depression has major health and social consequences. There is concern that general practitioners (GPs), who manage most depression in primary care, are reluctant to use assessment tools such as questionnaires that might improve diagnosis and management. A Cochrane systematic review has recommended qualitative exploratory studies to explore the impact of questionnaires on GP management. AIM: To identify effects of using mental health questionnaires on views of GPs managing depression, and how this might influence patient care. DESIGN: Qualitative, formative evaluation using principles of grounded theory in analysis of semi-structured interviews and focus groups before and after introducing the mental health questionnaire. SETTING: Four practices in South London. PARTICIPANTS: Twenty GPs, of whom four contributed to the pilot only, and 16 completed the main study. MAIN OUTCOME MEASURE: Identification of changes in GP views as a result of using the questionnaire with patients. RESULTS: Three themes emerged from analysis of GP views on managing depression: control and responsibility; the doctor-patient relationship, and support for the doctor. These were influenced by GPs' experience (years in practice), and perceived time spent dealing with depression (involvement). Use of the questionnaire enabled more experienced GPs to relinquish control, encourage patient involvement, and offer alternative sources of help. They felt less responsible for overall care. Less experienced, and less involved GPs found questionnaires supportive through increasing their confidence in asking difficult questions, and were encouraged to look for depression, which they might previously have avoided. CONCLUSIONS: Using mental health questionnaires helped GPs feel more confident in detecting and managing depression, and there was greater willingness to use questionnaires than found in previous studies. Most GPs sought reduced responsibility in ongoing care, using questionnaires to involve patients and pass responsibility to them or other agencies. PMID- 19807959 TI - The doctor-patient relationship in chronic fatigue syndrome: survey of patient perspectives. AB - BACKGROUND: When confronted with chronic fatigue syndrome (CFS), general practitioners (GPs) need to deal with diverse complaints. This may introduce a sense of powerlessness and frustration in the GP, which could possibly undermine the doctor-patient relationship. AIM: Our aim was to list the perspectives of patients with CFS regarding the medical encounter. METHOD: This was a questionnaire study of systematically selected patients presenting to a tertiary clinic specialising in CFS. A questionnaire was presented to every third patient attending the clinic. Statistical computations were performed using the SPSS statistical package. RESULTS: One hundred and seventy-seven patients completed the questionnaire. A diagnosis of CFS was made by a GP in 8% of the cases. In 31% of the cases the GP had experience with general CFS complaints, and 35% of the GPs showed experience in CFS. Only 23% reported sufficient knowledge to treat the condition. According to the patients surveyed, 35% felt that their GP had experience in dealing with CFS. CONCLUSIONS: The heterogeneity of CFS and the controversy surrounding this condition seemed to overwhelm GPs and strain the medical encounter. Patients with CFS seemed unsatisfied with the interaction with their doctor. Moreover, the results show that CFS is not addressed well by the medical community, and the failure to diagnose leads to a lack of empathetic care, with consequential loss of the capacity of the doctor to act as a healer. PMID- 19807960 TI - Lean experience in primary care. AB - The recent introduction and successful application of Lean Thinking in health is generating a belief that continuous improvement (a product of Lean) can be delivered in the context of health care, as long as key organisational principles are in place. This paper offers case studies to illustrate where Lean methods are being introduced into a primary care commissioning organisation and are delivering, as part of an integrated organisational approach that embraces Lean principles and improvement in the patient experience and eliminates the potential for error in patient care. PMID- 19807961 TI - Morton's neuroma: diagnostic accuracy, effect on treatment time and costs of direct referral to ultrasound by primary care physicians. AB - BACKGROUND: The first-line treatment for symptomatic Morton's neuroma in our hospital is a perineural ultrasound-guided injection of corticosteroid and local anaesthetic (USI). The NHS has recently implemented 18-week referral-to-treatment targets. When GPs specifically suggest a diagnosis of Morton's neuroma there are two referral pathways in our hospital: direct referral to radiology for USI (limited slots) or referral to the specialist foot and ankle clinic. Patients with less specific referral letters are also evaluated in clinic and referred for USI as appropriate. METHODS: A retrospective audit was performed reviewing referral letters from general practitioners (GPs) in 2005-2006. A comparison was made between the referral pathways for time-to-treatment (TTT), accuracy of GP diagnosis, and cost implications. RESULTS: In the directly referred group, the median TTT was 99 days, compared to 206 days for patients who went via a foot and ankle clinic (P < 0.001). Of 57 patients with a GP diagnosis of Morton's neuroma, 40 (70%) had the diagnosis confirmed on USI compared to 44 of 64 (69%) patients referred by a foot and ankle surgeon, showing no significant difference between the groups (P = 0.87). CONCLUSION: For patients with features highly suggestive of a Morton's neuroma, direct referral from primary care for USI had a similar accuracy to referral from a specialist hospital clinic and the time-to-treatment was significantly shorter. The mean waiting time of this group was within the 18 week government target without any changes to our current radiology protocols. PMID- 19807962 TI - The quality of primary care health centres in Buenos Aires, Argentina. AB - BACKGROUND: Primary healthcare strategies have been included in the health policies of many countries since the 1970s. The aim of this study was to evaluate the quality of care provided by primary care health centres (PCHCs) in Buenos Aires, Argentina. METHODS: We surveyed 20% of PCHCs in Buenos Aires State to investigate the structure, process and results of care. Variables related to 'structure' included human resources, qualifications and skills, geographical coverage and buildings. 'Process' variables were: shift handling, service organisation and demand planning. 'Outcome' or 'results' indicators included vaccination coverage, percentage of pregnant women attending before the 12th week, percentage of children consulting, and patient satisfaction. RESULTS: Our evaluation showed deficiencies in the structure and processes of care, and serious shortfalls in outcomes. Health teams in PCHCs lacked knowledge about the demography, health needs, and population statistics relating to their area of responsibility. Valid information was only available for the 13% of the population who sought help from the PCHC. As a result, there was low vaccination coverage (63.6%), low levels of child health surveillance (38.8%), and poor early detection of pregnancy (6.9%). CONCLUSION: Attention to these areas of deficiency could improve the quality of health care and outcomes for the communities of Buenos Aires. PMID- 19807963 TI - Personalisation of care: implications from two case studies for doctors and health services. PMID- 19807964 TI - Patient participation groups: the national picture. AB - This article brings together research carried out by the National Association for Patient Participation from 2005 to 2009. Its aim was to capture the views of GP surgeries that have patient participation groups (PPGs) as well as those who do not. It also studied the views of PPGs affiliated to the National Association for Patient Participation (NAPP) about their role, their organisation and the support that they require to maximise their success. With the exception of the online, UK wide members' survey, the research was carried out through postal surveys sent to more than 4000 English practices in three separate studies. The main results relate to the prevalence of PPGs, their geographical location, their activities, the attitudes of practices towards PPGs (especially those that do not have one) and the future needs of PPGs. PPGs are seen to play an important role in providing the practice with the patient perspective but they are active in several other areas, including health promotion, information provision, influencing commissioning, supporting other patients and fundraising (for their own needs as well as those of the practice). Three central challenges are highlighted, relating to (a) the most successful strategies to promote more PPGs; (b) the importance of support from primary care trusts (or their equivalent organisations); and (c) ways in which PPGs can increase their ability to represent the wider patient perspective. A future research agenda is also proposed that would evaluate the costs and benefits of PPGs across their broad range of activities. PMID- 19807965 TI - The rodent quarantine quagmire. PMID- 19807966 TI - Growth-associated changes in the periodontal bone and molar teeth of male rats. AB - Here we report quantitative data associating periodontal bone variables of young conventional rats with the growth process. The hemimandibles of male rats (IIM/Fm stock, 2 to 15 wk of age.) were excised and submitted to conventional morphologic, radiologic, and histologic evaluation. The length, area, or X-ray absorbance of various regions or structures was measured on digital images of radiographs by using an image-analysis program. The sum of periodontal bone areas undergoing resorption (interproximal + intraradicular) increased until 9 or 10 wk of age and decreased thereafter. Mineral accretion rates and mineral density asymptotes were not significantly different among molars. The mineral density of resorption areas in alveolar bone fitted sinusoidal kinetics, indicative of the 'instability' of the tissue due to its high metabolic activity. Mineral accretion rates and mineral density asymptotes were not significantly different among molars. The proportion of root length within alveolar bone exhibited a biphasic curve (minimum at 5 wk of age), due to differences in the growth rates of variables involved in its calculation (distance between the cementoenamel junction to the apex and height of the resorption areas). The distance between the cementoenamel junction and alveolar bone crest over time fitted a sigmoidal function with a point of inflection that did not differ significantly from that of body or mandible dry weight. In summary, the growth process appears to affect periodontal bone support and the distance between the cementoenamel junction and alveolar bone crest in male rats. PMID- 19807967 TI - Variation in clitoral length in rhesus macaques (Macaca mulatta). AB - Clitoromegaly in the neonatal period is an important morphologic sign that can be useful for sexual determination in aberrant cases. In rhesus monkeys, differentiation of the external genitalia occurs early during gestation (at 55 to 60 d) and is complete by approximately 80 d. Most of the critical steps in genital differentiation in primates occur prenatally. We sought to determine clitoral size in normal rhesus monkeys (Macaca mulatta) and possible effects of age and inheritance. Clitoral length was highly variable and had no relationship to fertility. Statistical evaluation revealed no association in the distribution of daughters with and without clitoris between mothers with and without clitoris. However, even when mated with several female monkeys, some male macaques produced primarily daughters without clitoris. PMID- 19807968 TI - Decreased blastocyst production in mice exposed to increased rack noise. AB - This study was conducted to investigate the possible effect of rack type on the blastocyst yield of mouse embryo donors. The first phase of the study consisted of housing some mice (group A) in a ventilated rack and others (group B) in a static rack in the same room for 3 d, followed by euthanasia for blastocyst collection and corticosterone assay. Parametric tests were used to compare groups. The number of blastocysts per donor was lower in group A (5.0 +/- 1.4 blastocysts) than group B (13.1 +/- 3.7 blastocysts). Mean noise was higher in the ventilated rack (80.4 dBC) than in the static rack (69.2 dBC). Serum corticosterone concentrations did not differ between groups. For the second phase of the study, a third group of mice (group C) was housed in a static rack without a ventilated rack in the same room. The noise level for group C was even lower (45.18 +/- 2.91 dBC), and the blastocyst count per donor (16.4 +/- 2.4) was higher than that of group B. The mean noise levels of empty ventilated and static racks differed significantly between groups for 10 different sound frequencies. Plotting mean blastocyst production against mean rack noise revealed a negative linear relationship with good strength of correlation. These results support the earlier observation that decreased blastocyst count occurs following housing of bred C57BL/6 donor mice in ventilated cages. PMID- 19807969 TI - Breeding and housing laboratory rats and mice in the same room does not affect the growth or reproduction of either species. AB - Few data exist regarding the effects of long-term housing of rats and mice in the same secondary enclosure. Historical reproductive and growth data were compared for colonies of mice and rats maintained in open-topped cages in either single species or dual-species barrier rooms. This analysis included reproductive parameters (litter size at birth, litter size at weaning, and pups missing at weaning) collected from 33 colonies of mice comprising 500 to 38,500 breeding females and 28 colonies of rats totaling 350 to 4,600 breeding females, and representative samples from 28 colonies of each species were analyzed for weight gain from weaning to adulthood. The presence or absence of the other species was not associated with statistically significant differences in weight gain or any of the reproductive parameters. These results suggest that breeding colonies of rats and mice of the same health status can be housed in the same room without a negative effect on the growth and reproduction of either species. PMID- 19807970 TI - Effect of chilling on the motility and acrosomal integrity of rat sperm in the presence of various extenders. AB - Studies were conducted to determine the effect of chilling on rat sperm and optimal components (extenders) to avoid chilling-induced injury. In the first experiment, the effects of chilling (at 4, 10, or 22 degrees C) on the motility and acrosomal integrity of epididymal sperm from 2 strains of rats (Sprague Dawley and Fischer 344, F344) were compared. In the second experiment, the motility of epididymal Sprague-Dawley rat sperm after exposure to extenders (HEPES-buffered Tyrode lactate, skim milk, lactose monohydrate, Tris-citrate, and TEST) and cooling and warming was determined. We tested the effects of supplementing base extender solutions with 20% lactose-egg yolk (LEY) alone or in combination with a commercial SDS-based paste (0.5%, v/v) in preventing chilling injury. The motility after each treatment was determined after both cooling and warming. In the third experiment, the motility of Sprague-Dawley rat sperm were compared after supplementing the base extenders with either 0.4 M permeating cryoprotective agent (CPA; glycerol, ethylene glycol, propylene glycol, or DMSO) or 0.1 M nonpermeating CPA (raffinose and sucrose) after cooling and warming. The results showed that chilling significantly reduced the motility-but not acrosomal integrity-of Sprague-Dawley and F344 sperm. Neither motility nor acrosomal integrity differed between Sprague-Dawley and F344 strains. The addition of LEY into each extender significantly prevented motility loss after chilling. These results will be useful during the preparation of optimal extenders and development of successful cryopreservation protocol for rat sperm. PMID- 19807971 TI - Euthanasia method for mice in rapid time-course pulmonary pharmacokinetic studies. AB - To develop a means of euthanasia to support rapid time-course pharmacokinetic studies in mice, we compared retroorbital and intravenous lateral tail vein injection of ketamine-xylazine with regard to preparation time, utility, tissue distribution, and time to onset of euthanasia. Tissue distribution and time to onset of euthanasia did not differ between administration methods. However, retroorbital injection could be performed more rapidly than intravenous injection and was considered to be a technically simple and superior alternative for mouse euthanasia. Retroorbital ketamine-xylazine, CO(2) gas, and intraperitoneal pentobarbital then were compared as euthanasia agents in a rapid time-point pharmacokinetic study. Retroorbital ketamine-xylazine was the most efficient and consistent of the 3 methods, with an average time to death of approximately 5 s after injection. In addition, euthanasia by retroorbital ketamine-xylazine enabled accurate sample collection at closely spaced time points and satisfied established criteria for acceptable euthanasia technique. PMID- 19807972 TI - Evaluation and refinement of euthanasia methods for Xenopus laevis. AB - The most common method of euthanasia for Xenopus species is by immersion in tricaine methane sulfonate solution (MS222). A wide range of doses of MS222 (0.5 to 5 g/L) have been recommended, but few reports describe dose-response testing, the time to loss of consciousness, or the reliability of euthanasia. The objective of this study is to evaluate the efficacy of immersing individual and groups of frogs in MS222 at concentrations ranging from 1 to 5 g/L for euthanasia and of 3 less-common methods: intracoelomic injection of MS222, intracoelomic injection of sodium pentobarbital with phenytoin, and ventral cutaneous application of benzocaine gel. Our results indicate that immersion for at least 1 h in a 5-g/L buffered solution of MS222, intracoelomic injection of 1100 mg/kg sodium pentobarbital with sodium phenytoin (equivalent to 0.3 mL solution per frog), or ventral cutaneous application of 182 mg/kg benzocaine (equivalent to a 2 cm x 1 mm of 20% benzocaine gel) is necessary to euthanize adult X. laevis and ensure complete cessation of the heartbeat without recovery. These doses are considerably higher than those previously recommended for this species. PMID- 19807973 TI - Interstrain differences in the development of pyometra after estrogen treatment of rats. AB - This case report describes the unanticipated development of pyometra in Brown Norway rats after treatment with estrogen. Sprague Dawley and Brown Norway rats were ovariectomized and randomly assigned to treatment groups (subcutaneous implantation of either a capsule containing 20 mg 17beta-estradiol or an empty capsule, as a control). After irradiation of only the right eye, the rats were followed for several months in an attempt to determine the effects of estrogen on radiation cataractogenesis and investigate potential strain differences in this phenomenon. However, all Brown Norway rats that received estradiol treatment developed pyometra, whereas none the Sprague Dawley or control Brown Norway rats did. This case demonstrates the potential adverse effects of exogenous estrogen therapy, which are strain-specific in the rat. Caution should be taken when designing estrogen-related experiments involving Brown Norway rats and other potentially sensitive strains. PMID- 19807974 TI - Enoxaparin treatment of spontaneous deep vein thrombosis in a chronically catheterized rhesus macaque (Macaca mulatta). AB - A chronically catheterized 14-y-old male rhesus macaque (Macaca mulatta) was reported for recurrent scrotal swelling. The scrotum was enlarged and warm to touch, and associated skin was noted to be lichenified on physical examination. The penis could not be extruded due to preputial swelling. Results from the following diagnostic tests were all unremarkable or within normal limits: scrotal aspirate, hematology, serum biochemistries, urinalysis, and radiography of the thorax, scrotum, and abdomen. Ultrasonography of lower extremities identified thrombi in bilateral iliac veins and left femoral vein. Collateral circulation surrounding the left femoral vein permitted some compensatory venous return. The left femoral vein of this animal had been catheterized approximately 2 mo before initial presentation. A coagulation panel revealed a positive D-dimer test, indicative of elevated levels of fibrin degradation products due to active thrombus breakdown. Enoxaparin sodium, a low-molecular-weight heparin for human use, was administered at 20 mg subcutaneously once daily for 10 d to treat occlusive venous thrombi. After enoxaparin treatment, the edema was greatly decreased. To achieve complete resolution, a second course of enoxaparin was administered 2 months after the first. Ultrasonography of the pelvic vasculature 6 mo after completion of therapy showed marked thrombus resolution, allowing for bilateral patency in the iliac and femoral veins. Follow-up evaluation revealed that D-dimer values were negative as well. This case demonstrates the novel application of the human medication enoxaparin to treat clinical signs of deep vein thrombosis in a chronically catheterized rhesus macaque. PMID- 19807975 TI - Nasal polyp cell populations and fungal-specific peripheral blood lymphocyte proliferation in allergic fungal sinusitis. AB - BACKGROUND: Allergic fungal sinusitis (AFS) is considered a different disease from other polypoid chronic rhinosinusitis diseases (CRS) with eosinophilic mucus (EM) termed eosinophilic mucus chronic rhinosinusitis (EMCRS). To substantiate this, studies on cellular responses to fungi and sinus mucosal inflammatory cell populations in AFS and other EMCRS diseases are required. This study was designed to examine polyp inflammatory cell populations and peripheral blood fungal specific T-cell responses in AFS, other EMCRS subgroups (defined later), and polypoid CRS without EM. METHODS: A prospective study was performed. Clinical characteristics, including CRS symptoms, sinus computed tomography (CT) scans, allergy status, intraoperative endoscopy, presence of EM, and fungal culture results were used to define patient groups. Polyps and peripheral blood were examined for populations of eosinophils, lymphocytes (CD4+, CD8+ T cells, natural killer cells, and B cells), and neutrophils using immunohistochemistry, cytospin preparations and flow cytometry. Fungal-specific peripheral blood lymphocyte proliferation was examined in AFS patients, other EMCRS patients, CRS patients, and controls. RESULTS: There was no significant difference in the percentage of cell populations and fungal-specific lymphocyte proliferation between AFS and other EMCRS diseases. However, AFS and other EMCRS polyps had a higher percentage of eosinophils and CD8+ T cells whereas CRS polyps had higher CD4+ T cells. Fungal-specific lymphocyte proliferation was significantly greater in AFS and other EMCRS patients regardless of fungal allergy, whereas in CRS and controls, higher proliferation was observed in fungal-allergic individuals. CONCLUSION: These findings question the basis for differentiating AFS from other EMCRS diseases based on fungal allergy and fungi in EM. Fungal-specific cellular response was present in AFS and other EMCRS diseases, different from that associated with fungal allergy, suggesting a nonallergic fungal immune response. Increased CD8+ T cells in EMCRS polyps signify a different type of inflammation to CRS that may be driven by CD8+ T cells. PMID- 19807976 TI - Detection of Staphylococcus aureus in nasal tissue with peptide nucleic acid fluorescence in situ hybridization. AB - BACKGROUND: Staphylococcus aureus (SA) in the nose can be a simple colonizer but also may create an intramucosal reservoir causing recurrent infections or can be a specific immune modulator through superantigenic mechanisms. Because the colonization rate of SA is high, but immunologic reactions causing chronic disease are less frequent, the purpose of this study was to identify the presence of intramucosal SA in healthy subjects and in patients with chronic rhinosinusitis (CRS) and to eventually relate those to the specific immunologic changes due to SA enterotoxins. METHODS: Nasal tissue was collected in 40 subjects (9 controls, 21 CRS patients with [CRSwNP], and 10 CRS patients without nasal polyps [CRSsNP]). Tissues were homogenized, and mediators and specific IgE antibodies against SA enterotoxins (SAE-IgE) were measured using the UniCAP system. The tissue was analyzed for the presence of SA by the peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) technique (AdvanDx), and a semiquantitative scoring system was applied. Mann-Whitney exact test was used for statistical analysis. RESULTS: SA in the mucosal tissue was detected in a higher quantity among CRSwNP subjects with aspirin exacerbated respiratory disease (AERD) versus controls and CRSsNP (p=0.03). Among CRSwNP patients, Th2 markers (eosinophil cationic protein, p=0.006, and total IgE, p=0.004) were increased related to the SAE-IgE status but not related to the presence of SA in the tissue. CONCLUSION: This study describes the detection of SA within nasal tissue using the PNA-FISH technique. The presence of SA in the submucosa did not correlate with the amplification of the Th2-related inflammation typically found in CRSwNP patients, but this reaction is dependent on the formation of SAE-IgE within mucosal tissue. We also show, for the first time, that submucosal SA is a prevalent finding in CRSwNP patients with AERD. PMID- 19807977 TI - Eosinophil degranulation patterns in nasal polyposis: an ultrastructural study. AB - BACKGROUND: Eosinophils are possibly the most important inflammatory cells in the pathogenesis of rhinosinusitis with nasal polyposis. Eosinophil degranulation is the mechanism by which these cells exert their inflammatory action. Knowledge of eosinophil state and degranulation mode therefore may help us to better understand this disease. A study is made of eosinophil state and degranulation mode using transmission electron microscopy (TEM), attempting to establish correlations with certain clinical variables considered to be of importance in patients with nasal polyposis. METHODS: A prospective TEM study was made to examine 582 eosinophils under 5000x magnification, classifying them according to their state and degranulation mode. The cells originated from 36 cases of nasal polyposis and were catalogued according to the clinical-radiological presentation of the disorder, the presence of asthmatic disease, or acetilsalicilic acid (ASA) triad syndrome (nasal polyposis, asthma, and nonsteroidal anti-inflammatory drug intolerance), and the degree of nasal tissue eosinophilia of the case of origin. RESULTS: A total of 30.75% of the eosinophils were inactive, 41.75% exhibited piecemeal degranulation (PMD), 27.5% exhibited cytolysis, and 0.34% were in apoptosis. The degranulation mode was significantly correlated to the clinical and histological parameters studied. Thus, cytolysis tended to be less pronounced and PMD greater, in the cells from cases with intense eosinophilia or severe polyposis (high clinical stage and ASA triad). CONCLUSION: Cytolysis and PMD are the principal degranulation modes of eosinophils in nasal polyposis-apoptosis being very infrequent. Nasal polyposis shows a correlation between eosinophil degranulation mode and the clinical and radiological stage and the degree of tissue eosinophilia of the case of origin. PMID- 19807978 TI - Dynamic nasal infrared thermography in patients with nasal septal perforations. AB - BACKGROUND: Nasal obstruction is a typical symptom in patients with nasal septal perforations. Rhinomanometry and acoustic rhinometry are not reliable in these cases because the perforations generate incorrect results. Infrared thermography camera (ITC) systems allow contact-free intranasal recordings of the nasal surface temperature and the semiquantification of nasal airflow. The aim of this study was to perform contact-free temperature measurements of the nasal vestibular surface by application of ITC systems in patients with septal perforations to investigate the disturbed intranasal heat exchange and nasal airflow. METHODS: The surface temperature profiles within the nasal vestibules of healthy volunteers (n=10) and patients with septal perforations (n=3) were recorded with an ITC during several breathing cycles. Thermal images were taken (60/s) displaying the surface temperature in degrees centigrade corresponding to a color scale. RESULTS: The temperature recordings showed a disturbed intranasal heat exchange during inspiration and expiration in patients with septal perforations in comparison with healthy subjects. A reduced and irregular inspiratory cooling of the entire surface within the nasal vestibules visualizes a reduced and disturbed airflow volume. CONCLUSION: The study was able to prove the feasibility of intranasal temperature recordings of the surface with an ITC system in patients with septal perforations. Contrary to rhinomanometry and acoustic rhinometry, thermography cameras can be applied to examine airflow in patients with septal perforations. The detected reduced cooling of the surface during inspiration might be a possible explanation for the patients' feelings of nasal obstruction. PMID- 19807979 TI - Nasal physiological reactivity of subjects with nonallergic rhinitis to cold air provocation: a pilot comparison of subgroups. AB - BACKGROUND: Noninfectious nonallergic rhinitis (NINAR) is characterized by self reported hyperreactivity to nonspecific physical or chemical stimuli. The relationship between these two classes of triggers is not well established, however. We compared NINAR subjects with predominantly physical or chemical triggers versus normal controls with respect to subjective (symptomatic) and objective (obstructive) responses to cold, dry air challenge. METHODS: We studied 14 NINAR subjects and 10 normal controls. Exposures consisted of 15 minutes of cold dry air (0 degrees C/5% RH) or warm moist air (25 degrees C/50% RH) on two separate days a week apart. Subjects rated symptoms using visual analog scales and had their nasal airway resistance measured at baseline, immediately after, and at 15-minute intervals for 1 hour postexposure. RESULTS: The majority of NINAR subjects reported physical triggers as more troublesome than chemical. Immediately postprovocation, the mean net proportional change in nasal airway resistance from baseline was +0.18 in NINAR (physical), +0.05 in NINAR (chemical), and -0.01 in control subjects (NS). However, a pooled linear regression by number of physical triggers (0-5) revealed a 7.5% increase in cold air-induced nasal airway resistance per trigger reported (p<0.05). Similarly, raising the criterion number of physical triggers from >or=1 to >or=2 also distinguished NINAR subjects from controls in a bivariate analysis. CONCLUSION: Either considering self-reported physical triggers as a continuous scale (0-5) or requiring more physical triggers (>or=2 rather than >or=1) to define NINAR successfully predicts objective nasal reactivity to cold air provocation. PMID- 19807980 TI - Cartilage outgrowth in fibrin scaffolds. AB - BACKGROUND: Fibrin glue has been a favorable hydrogel in cartilage tissue engineering, but implantation of chondrocyte-fibrin suspensions have resulted in volume loss. In this study, human septal cartilage chips were seeded onto a fibrin scaffold, and cellular proliferation and production of cartilaginous extracellular matrix (ECM) were evaluated. METHODS: Human septal cartilage was diced into cartilage chips and encased with and without fibrin glue. Four conditions were initially tested for DNA content and glycosaminoglycan (GAG) production: (1) control medium in tissue culture, (2) control medium with fibrin glue, (3) collagenase-supplemented medium in tissue culture, and (4) collagenase supplemented medium seeded in fibrin glue. Cartilage chips cultured in collagenase-treated medium were then seeded onto either cell culture plates, suspended in alginate, or mixed with fibrin. Cellular proliferation, GAG production, and histochemistry were evaluated. RESULTS: Fibrin preparations increased cellular proliferation and DNA content. GAG levels were highest in collagenase-treated samples encased in fibrin. Cartilage chips treated with collagenase showed increased cellular proliferation in the fibrin preparations compared with preparations without fibrin. GAG increased with the addition of fibrin when compared with explant. Histochemistry revealed increased GAG accumulation in the regions between the cartilage chips with the addition of fibrin. CONCLUSION: Adding fibrin glue to collagenase-treated cartilage chips results in increased proliferation and maintains ECM production and, therefore, may facilitate generation of cartilaginous tissue for use in reconstructive surgery. PMID- 19807981 TI - Effect of chronic rhinosinusitis on liver transplant patients. AB - BACKGROUND: The use of immunosuppressant after liver transplantation makes transplant recipients susceptible to infections. Most infections that can alter mortality after liver transplantation are wound infections, urinary infections, and pneumonias. There is no evidence, however, that chronic rhinosinusitis can alter mortality in patients awaiting liver transplantation. We have therefore assessed the association of rhinosinusitis with mortality and prognosis after liver transplantation. METHODS: The clinical records of 996 patients who received liver transplants between January 1995 and March 2005 were reviewed and the collected data were analyzed. RESULTS: Of the 996 patients who received liver transplants between January 1995 and March 2005, 28 (2.8%) had pretransplant rhinosinusitis. Of the latter, 5 patients were treated medically, 1 patient had endoscopic sinus surgery, and 22 patients had no treatment before liver transplantation. Untreated rhinosinusitis before liver transplantation was associated with aggravated rhinosinusitis after transplantation but did not contribute to an increase in infectious mortality or overall mortality rate. CONCLUSION: Pretransplant chronic rhinosinusitis does not contribute to mortality in patients undergoing liver transplantation. PMID- 19807982 TI - Computed tomography findings in sinonasal Wegener's granulomatosis. AB - BACKGROUND: Wegener's granulomatosis (WG), an autoimmune disease, is intimately associated with the sinonasal tract, with involvement reported in 85% of patients during the course of the disease process. The objective of this study was (1) to describe Lund-Mackay (L-M) scores, (2) to delineate patterns of neo-osteogenesis and bony erosion, and (3) to analyze the impact of surgery on the computed tomography (CT) findings of WG patients. METHODS: A retrospective review was performed on 74 patients with WG presenting to a tertiary care referral center. CT analysis was performed and graded by two independent reviewers. RESULTS: The mean age was 53 years with a male/female ratio of 0.6:1. The average L-M score was 10.0. Neo-osteogenesis was evident in 78% of the patients with overall average neo-osteogenesis score of 4.2 (range, 0-16). Bony erosion was noted on imaging in 62% of patients with overall average score for bony erosion of 2.0 (range, 0-8). Patients having undergone previous sinus surgery compared with no previous surgery had statistically significant elevation of overall L-M, bony erosion, and neo-osteogenesis scores (p=0.024, 0.0009, and 0.0015, respectively). CONCLUSION: CT imaging in WG patients shows elevated L-M scores and evidence of bony erosion and neo-osteogenesis. Furthermore, surgical manipulation in WG patients is associated with increased bony abnormalities and greater elevations of L-M scores, possibly because of worsening vasculitis and/or inflammation. The presence of concurrent neo-osteogenesis and bony destruction of the paranasal sinuses should raise clinical suspicion of WG in patients presenting with symptoms of chronic rhinosinusitis. PMID- 19807983 TI - Comparison between three-dimensional and triplanar computed tomography imaging of the frontal recess. AB - BACKGROUND: Despite advances in endoscopic surgical techniques, management of frontal sinus disease remains challenging. Much of this is related to the complex nature of frontal recess anatomy. A thorough understanding of frontal recess anatomy is paramount for the safety and success of frontal sinus surgery. Three dimensional (3D) computed tomography (CT) may allow surgeons to obtain a more complete preoperative assessment of frontal recess anatomy. The purpose of this study was to determine if reconstructed 3D CT images as an adjunct to conventional triplanar imaging provide additional information regarding the frontal recess anatomy. METHODS: A prospective study was performed. Two otolaryngologists reviewed the CT scans of 25 patients referred for routine paranasal sinus disease. The findings from review of the triplanar CT images were compared with the findings from review of the 3D reconstructions. Each study was assessed for (1) frontoethmoidal cells, (2) agger nasi cell, (3) subrabullar and frontal bullar cells, (4) intersinus septal cell, (5) superior uncinate process attachment site, and (6) and frontal sinus outflow tract. The examiners rated the usefulness of each study to identify each of the aforementioned anatomic subsites using a modified 5-point Likert scale. RESULTS: Intersinus septal cells, supraorbital ethmoid cells, and the anterior-posterior dimension of the frontal sinus outflow tract were better defined on the reconstructed 3D CT images. CONCLUSION: Three-dimensional CT is a useful adjunct to the conventional triplanar studies for the evaluation of frontal sinus and recess anatomy. This technique can define certain anatomic variants more effectively than 2D multiplanar reconstructed images. PMID- 19807984 TI - Biofilms and mucosal healing in postsurgical patients with chronic rhinosinusitis. AB - BACKGROUND: Patients with chronic rhinosinusitis (CRS) often remain symptomatic after technically proficient functional endoscopic sinus surgery. Current hypothesis indicates biofilms may contribute to the persistence of infection. However, few studies showed biofilms in postoperative patients. This study was designed to identify bacterial biofilms on postoperative mucosa, as well as to investigate the healing of sinus mucosa after surgery. METHODS: After intraoperative mucosa was obtained for assessment of biofilms, 27 patients were followed up for 6 months. Postoperative medications and symptoms were recorded. As indicated by endoscopic evaluation, biopsy specimens of postoperative edema, scar, or adhesion were obtained. Samples were prepared for scanning electron microscopy (SEM) and hematoxylin and eosin (H&E) staining. RESULTS: Fifteen postoperative samples were taken from the 20 patients with intraoperative biofilms. Under SEM, postoperative biofilms were identified in 4/6 scar samples and 5/9 edema samples. There was no significant difference in biofilm presence between samples of scar and edema. Microcolonies were also identified on postoperative scar under H&E staining. The presence of intraoperative and postoperative biofilms was correlated with the severity of preoperative Lund MacKay computed tomography score and postoperative Lund-Kennedy endoscopic score. Compared with intraoperative samples, postoperative samples from the same nine patients significantly recovered from ciliary damage, metaplasia, and basement membrane thickness. Postoperative cultures were positive in samples with and without postoperative biofilms. CONCLUSION: Biofilms persist after treatment, and may cause the unfavorable outcomes of surgery for CRS. The mucosa with biofilms can recover after surgery. Apparent bacterial plaque can be identified by H&E staining. PMID- 19807985 TI - Double-blind, placebo-controlled trial of reformulated azelastine nasal spray in patients with seasonal allergic rhinitis. AB - BACKGROUND: Azelastine nasal spray is a topical antihistamine with a distinctive taste that may be objectionable to some patients. The primary objectives of this clinical trial were (1) to determine if a reformulated azelastine nasal spray (Astepro) with sucralose as a taste-masking agent provides comparable efficacy to the original formulation (Astelin) and (2) to evaluate dose-response relationships between groups. METHODS: Eight hundred thirty-five patients with seasonal allergic rhinitis were randomized to six treatment groups: (1) original azelastine nasal spray, 1 spray/nostril b.i.d.; (2) reformulated azelastine, 1 spray/nostril b.i.d.; (3) placebo, 1 spray/ nostril b.i.d.; (4) original azelastine nasal spray, 2 sprays/nostril b.i.d., (5) reformulated, 2 sprays/nostril b.i.d.; and (6) placebo, 2 sprays/nostril b.i.d. The primary efficacy variable was the change from baseline to day 14 in total nasal symptom score (TNSS) consisting of runny nose, sneezing, itchy nose, and nasal congestion. RESULTS: Original azelastine nasal spray and the reformulated spray produced comparable improvements in the TNSS at both dosages. There was a dose related difference in TNSS comparing the 1- and 2-spray dosages. The percentage changes from baseline in the TNSS in the 2-sprays/nostril dosage groups were 27.9% (p<0.001) with the reformulated nasal spray, 23.5% (p<0.01) with the original formulation, and 15.4% with placebo. The incidence of bitter taste was 7% with the reformulated spray and 8% with the original at the 2-sprays/nostril dosage. CONCLUSION: The results of this study showed efficacy both with original azelastine nasal spray and with the reformulated nasal spray and a clear dose response difference between the 1- and 2-spray dosages. PMID- 19807986 TI - Nasoseptal flap reconstruction of high flow intraoperative cerebral spinal fluid leaks during endoscopic skull base surgery. AB - BACKGROUND: Over the past 10 years, significant anatomic, technical, and instrumentation advances have facilitated the exposure and resection of intradural lesions via a fully endoscopic expanded endonasal approach (EEA). The vascularized nasoseptal flap (based on the posterior nasoseptal artery) has become our primary endoscopic reconstructive technique. The goals of this study are to prospectively evaluate the nasoseptal flap and high-risk cerebral spinal fluid (CSF) leak variables. METHODS: Prospective evaluation was performed of EEA patients with intraoperative high-flow leaks (either a cistern or ventricle open to nasal cavity during tumor dissection) who underwent nasoseptal flap reconstruction. RESULTS: Seventy consecutive nasoseptal flaps for high-flow intraoperative leaks were evaluated prospectively by the primary author. Twelve risk factors were then graded at the time of the operations and correlated to CSF leak outcomes. The overall postoperative CSF leak rate was 5.7% (4/70). All four postoperative leaks were successfully managed with endoscopic repair and CSF diversion. A multivariate analysis of all 12 risk factors is detailed. Pediatric patients, large dural defects, and radiation therapy were noted to be factors in reconstructive failure. One flap death occurred in a patient with prior surgery and proton therapy, this leak was managed with a temporoparietal flap and endonasal repair. CONCLUSION: The nasoseptal flap is an excellent anterior skull base reconstructive technique. Patients with high-flow intraoperative CSF leaks had a 94% successful reconstruction rate. Patients with skull base proton radiation therapy are at higher risk for flap failure and preparation for nonradiated tissue reconstruction should be discussed with the patient. PMID- 19807987 TI - Inferior turbinate pedicle flap for endoscopic skull base defect repair. AB - BACKGROUND: Endoscopic skull base reconstruction (ESBR) is an important advance in the management of skull base defects. Large dural defects usually require the use of pedicled mucosal flaps for successful repair and prevention of cerebral spinal fluid leak. Planning for pedicled flaps is important because raising the flap is often required before tumor removal or initial surgical access. The potential utility of the inferior turbinate pedicled flap (ITPF) in ESBR is assessed. METHODS: The ITPF was raised in nine cadaver heads. The most anterior extent reached on the anterior cranial fossa (ACF) and inferior limit on the posterior cranial fossa (PCF) were recorded with image-guided surgery. Measurements were calculated as a percentage of ACF or PCF length from easily reproducible measurements from preoperative radiology. The width was defined as the widest measurement in the distal third. The length-to-height ratio of the skull base was also assessed as a confounding factor. RESULTS: Mean length for the ITPF was 54.0+/-4.9 mm with a width of 22.1+/-3.7 mm. The mean reach was 112+/-21% (range, 90-150%) of the PCF. The ACF was less accessible with a mean reach of 67+/-9.9% (range, 52-84%). The skull base proportions did not have a strong association on utility (p=0.74 and 0.29). CONCLUSION: ITPF presents one option for ESBR of larger skull base defects. It has several limitations with access to the anterior ACF. The ITPF may be an excellent salvage flap when previous septectomy has been performed or with septal involvement by tumor. Careful preoperative assessment of potential defect can estimate the reach of the ITPF. PMID- 19807988 TI - KTP/532 YAG laser treatment for allergic rhinitis. AB - BACKGROUND: The purpose of this study was to investigate the efficacy of the KTP/532 YAG laser to reduce nasal congestion and discharge in patients with allergic rhinitis. METHODS: Forty-eight patients with symptoms of allergic rhinitis were treated with the KTP/532 laser. All had positive skin tests for common allergens. Treatments were provided on an ambulatory basis in one to three sessions under local anesthesia with lidocaine nose spray. Outcome was determined by daily symptom reports and regular endoscopy examination and interviews for 12 months. RESULTS: Treatment was very well tolerated. There were no major side effects. At examination after 1 year, nasal obstruction was improved in 69% and nasal discharge in 40% of cases. CONCLUSION: The KTP/532 YAG laser is effective for the treatment of nasal obstruction and discharge. Comparison with other techniques showed it to be the most effective in reducing nasal discharge. It can be done as an office procedure and does not damage the nasal mucous membrane. The KTP/532 YAG laser is effective as an additional treatment for patients refractory to medication. PMID- 19807989 TI - Changes in the maxillary sinus volume and the surgical outcome after the canine fossa puncture approach in pediatric patients with an antrochoanal polyp: results of a minimum 3-year follow-up. AB - BACKGROUND: An antrochoanal polyp (ACP) originates from the maxillary antrum and simple excision results in high recurrence rates. Canine fossa puncture (CFP) has been proposed as an alternative method of accessing the entire maxillary antrum. To investigate the long-term effects of the CFP approach on changes in the maxillary sinus volume in pediatric patients with an ACP, we compared the sinus volume on the operated and normal sides, using preoperative computed tomography (CT), and postoperative CT performed at least 3 years after the procedure. The surgical outcomes were also assessed using endoscopic and CT findings. METHODS: Seven patients met the inclusion criteria and the ACP was removed via the CFP approach. After a mean follow-up period of 43.9 months, we compared the changes in the maxillary sinus volume between the operated and normal sides, using the pre- and postoperative CT data. RESULTS: No contractures or decrease in the maxillary sinus volume on the diseased side were observed in any of the patients on postoperative CT, and the average volume of both maxillary sinuses was greater on postoperative CT compared with the preoperative scans. None of the patients showed evidence of recurrence on the endoscopic and CT examinations. CONCLUSION: CFP did not affect the maxillary sinus volume in pediatric patients with an ACP and led to a successful surgical outcome in all of the patients. Based on these results, we recommend CFP as a safe, effective method for the treatment of ACP. PMID- 19807990 TI - Rhinoplasty to correct nasal deformities in postseptoplasty patients. AB - BACKGROUND: Although not so common, septoplasty carries the risk of inducing nasal deformity such as saddle nose and deviated nose. Surgical correction for those deformities is a formidable task to deal with, because the septal cartilage framework was already weakened due to previous procedure. This study was designed to examine various causes and effective techniques of rhinoplasty for correction of nasal deformities after septoplasty. METHODS: Subjects comprised 19 patients who developed nasal deformities after septoplasty. Pre- and postoperative photographs were analyzed by two rhinoplastic surgeons. Anthropometric measurements of the external nose before and after surgery were made using frontal and lateral photographs to objectively evaluate outcomes. RESULTS: Intraoperative analysis showed that 5 patients had weak L-struts and 13 patients had displaced caudal septa. Septum was reconstructed using costal cartilage in 13 cases. Spreader grafting was performed in 13 patients and caudal septal extension grafts were performed in 10 patients. All but 1 patient had successful functional and esthetic outcomes. In the saddle nose group, all parameters except nasolabial angle were significantly improved. In the deviated nose group, curved deviated angles were completely corrected, whereas the linear deviated angle was not. CONCLUSION: Loss of caudal septal support appears to be a major reason for nasal deformity after septoplasty. The best surgical approach for septal cartilage framework reconstruction appears to involve the use of bilateral spreader or septal extension grafts using costal cartilage. PMID- 19807991 TI - The effect of functional endoscopic sinus surgery on nasal spray distribution to the middle meatus in patients with rhinosinusitis. AB - BACKGROUND: Because the principle behind functional endoscopic sinus surgery (FESS) recognizes the middle meatus as essential for the control of the disease, the effect of the operation in the distribution of drugs in the same area should be of similar importance. This study was designed to clarify whether nasal drug delivery is improved in patients after FESS. The study involved a prospective assessment within subject comparison. The subjects were 20 adult (>18 years old) patients on the waiting list for FESS. It was conducted within two teaching hospitals in the East of Scotland. METHODS: A novel method was used, positioning a neurosurgical patty in the middle meatus and assessing blue dye nasal spray absorption on a 4-point scale. RESULTS: A significant improvement was observed in the distribution of the indicator in the middle meatus postoperatively. Seventy percent of the patients showed improvement in the drug distribution after FESS. The median result for absorption score was 2 preoperatively (<50%) and 3 postoperatively (>50%). The difference was statistically significant (p<0.001). CONCLUSION: The distribution of nasal drugs is improved in the middle meatus after FESS. This can have important clinical applications that can benefit patients in otolaryngology as well as other disciplines. PMID- 19807996 TI - Contact dermatitis from acrylics in a histology laboratory assistant. AB - The use of acrylics has expanded enormously, resulting in a vast range of products for both occupational and non-occupational purposes. Acrylics reported to cause allergic contact dermatitis in histology technicians are 2-hydroxyethyl methacrylate and 2-hydroxyethel acrylate. PMID- 19807992 TI - Impaired mucosal healing and infection associated with Staphylococcus aureus after endoscopic sinus surgery. AB - BACKGROUND: Differentiating normal nasal discharge and postoperative crust from infection after endoscopic sinus surgery (ESS) can be difficult. We investigated whether bacteria cultured at operation was predictive for postoperative mucosal infection-defined as frank pus, thick mucus, and/or abnormal crusting seen on endoscopic examination associated with positive microbiology. METHODS: The records of 48 patients with chronic rhinosinusitis (CRS) with infection at the time of ESS were retrospectively reviewed. Results of intraoperative cultures were compared with those taken at the time of postoperative mucosal infection. RESULTS: Fourteen of 16 patients (87.5%) with intraoperative infection with Staphylococcus aureus progressed to postoperative mucosal infection with S. aureus, whereas patients who cultured "other" bacteria intraoperatively progressed to postoperative mucosal infection in 6/19 cases (31.6%), with S. aureus cultured in only 3/19 cases (15.8%; p=0.0001). CONCLUSION: S. aureus infection at ESS predicts for abnormal, S. aureus-associated mucosal healing and infection post-ESS. Although a prospective trial is warranted, these findings suggest a future role for aggressive anti-S. aureus therapy peri- and/or postoperatively in patients who culture positive for this organism to improve postsurgical outcomes. PMID- 19807998 TI - A mechanism-based classification of dermatologic reactions to biologic agents used in the treatment of cutaneous disease: Part 2. AB - Biologic therapies are an efficacious new method of controlling a number of chronic conditions. Data regarding these medications continues to emerge, giving clinicians a greater understanding of their side effects profiles. The biologic agents used in dermatology, particularly the tumor necrosis factor-alpha inhibitors, have a number of varied dermatologic side effects. In this two-part article, we perform a review of literature regarding cutaneous side effects of infliximab, etanercept, adalimumab, rituximab, efalizumab, and alefacept. In this second part, we discuss injection site reactions, infusion reactions, vasculitis, drug-induced lupus erythematosus, psoriasiform lesions, rebound phenomenon, eczema, atopic dermatitis, and hypersensitivity reactions. PMID- 19807999 TI - Compilation of international standards for patch testing methodology and allergens. AB - BACKGROUND: Recommendations for patch testing methodology and allergens differ between contact dermatitis groups worldwide. Currently, there is no compilation of these recommendations. OBJECTIVE: This study aims to summarize the current recommendations for patch testing methodology and allergens. METHODS: Data was collected through a literature search and by contacting representatives from 24 known contact dermatitis groups. RESULTS: Thirteen groups responded to our inquiry. As expected, recommendations for allergens and methodology varied between different groups; however, most groups based their recommendations on the European baseline series. We have compiled tables of methodologies and commonly tested allergens. CONCLUSION: The differences between groups underscore the need for further international collaboration among contact dermatitis groups. PMID- 19808000 TI - A localized flare of dermatitis may render patch tests uninterpretable in some patients with recently controlled widespread dermatitis. AB - BACKGROUND: Patch testing rarely is confounded by localized dermatitis induced in the area being tested (usually the back). Its occurrence renders the interpretation of patch tests impossible. OBJECTIVE: To review our experience of the circumstances in which this phenomenon occurs during patch testing. METHODS: We retrospectively reviewed patients with this phenomenon who underwent patch testing from January 1, 2002, through June 30, 2006. RESULTS: Of the 3,569 patients tested, 12 (0.34% [9 men and 3 women]) had development of this phenomenon. All patients previously had recent widespread dermatitis that was suppressed temporarily with topical corticosteroids and wet dressings at the time of patch testing. The period between control of the dermatitis and the initiation of patch testing was less than 1 week for all patients. Three patients (25%) had recently discontinued therapy with systemic corticosteroids (less than 1 week earlier). CONCLUSION: In patients with irritable skin either immediately after widespread dermatitis is controlled or after the cessation of systemic corticosteroid treatment, a flare of dermatitis induced by patch testing may render patch tests unreadable and therefore uninterpretable. To avoid this confounding occurrence, a waiting period between control of widespread dermatitis and initiation of patch testing is advised. PMID- 19808001 TI - Patch-testing while on systemic immunosuppressants. AB - BACKGROUND: Occasionally, the need arises to patch-test patients while they are on immunomodulators. Little is known about how these systemic agents affect the results of patch testing. OBJECTIVE: To present data on 11 patients who underwent patch testing while under the effects of immunosuppressants. METHODS: Retrospective chart reviews were performed on 11 patients who underwent patch testing while they were taking various systemic immunosuppressants or within 48 hours of cessation of various systemic immunosuppressants. RESULTS: Patients had been taking prednisone (n = 6), cyclosporine (n = 2), combination cyclosporine and prednisone (n = 1), mycophenolate mofetil (n = 1), and infliximab (n = 1) up to 48 hours prior to and/or during patch testing. Seven patients showed at least one strong (++) or extreme (+++) patch-test reaction. Three patients had at least one weak (+) reaction. One patient showed only questionable reactions. The patient on mycophenolate was eventually retested while off immunosuppressants and showed strong clinically relevant patch-test reactions. Overall, 8 of the 11 patients reported some improvement in their dermatitis, including all the patients with strong or extreme reactions. CONCLUSION: While it is optimal for patch testing to be performed when patients are off immunosuppressants, immunosuppressive therapies should not be an absolute contraindication to patch testing. PMID- 19808002 TI - Methylphenidate patch-test protocol and irritancy threshold determination in healthy adult subjects. AB - BACKGROUND: The methylphenidate transdermal system (MTS) (Daytrana [Shire Pharmaceuticals Ireland Ltd., Wayne, PA]) has been shown to be well tolerated and effective; however, skin reactions, typically redness and itching, have also been noted with MTS use. OBJECTIVE: The purpose of this study was to determine the irritancy threshold of methylphenidate (MPH), the active ingredient in the MTS, in healthy adults for use in subsequent patch testing. METHODS: This was an open label single-site pilot study. Eligible subjects had MPH (six different concentrations in two vehicles) patch tests applied to the paraspinous region of the upper back. Subjects returned after 48 hours and 96 hours for investigator assessments of each reaction. RESULTS: All enrolled subjects (n = 20) completed the study. There were no definite positive reactions in any subject at any test concentration. Overall, doubtful (macular erythema) reactions were noted on 12 sites at the 48-hour reading and on 6 sites at the 96-hour reading. With both aqueous and petrolatum vehicles, more doubtful reactions were noted at the lower MPH test concentrations. CONCLUSION: When performing patch testing to confirm possible allergic contact dermatitis from topical MPH, several test concentrations in the low-to-middle range (such as 0.1%, 1%, and 10%) prepared in petrolatum are advisable. PMID- 19808003 TI - Direct observed "soak and smear" therapy for severe eczematous dermatitis in the combat setting. AB - BACKGROUND: Eczematous dermatitis is difficult to control in a combat setting and may result in a service member's early return to the United States. We report on seven patients with presumed eczematous dermatitis for whom traditional treatments had failed. OBJECTIVE: To evaluate the effectiveness of "soak and smear" in eczematous dermatitis. METHOD: The patients were treated with 3 days of direct observed soak and smear therapy with topical steroids. RESULTS: All seven patients significantly improved with treatment, showing a decrease in investigator global assessment, patient self-assessment, and pruritus scores. CONCLUSION: This study demonstrates that direct observed soak and smear therapy is an effective tool with which to rapidly improve severe eczema. Additionally, this method can be used in the combat environment. PMID- 19808004 TI - Repeated mechanical trauma to the hands: the use of anti-impaction gloves for treatment and return to work. AB - BACKGROUND: Repeated mechanical trauma to the hands can result in frictional hand dermatitis (FHD) and (in some cases) hyperkeratotic hand dermatitis (HHD), forms of irritant contact dermatitis. These conditions can be chronic and debilitating and are often refractory to many therapies. Most case reports of FHD describe avoidance of frictional trauma as a management solution. This is the first study to examine anti-impaction gloves as a treatment for FHD. OBJECTIVE: To determine whether anti-impaction gloves are effective in helping patients with FHD and HHD return to work. METHODS: We describe a small case series of a subset of patients who presented to our occupational contact dermatitis clinic between January 2004 and June 2008 with either HHD or FHD. Of the 11 patients evaluated, 9 had a diagnosis of FHD and 2 were diagnosed with HHD. They were all treated with anti impaction gloves, tazarotene cream (Allergan, Inc., Irvine, CA), and clobetasol propionate ointment (Taro Pharmaceuticals Inc., Brampton, Ontario, Canada) and were followed over the long term to determine the impact of treatment on their ability to maintain their jobs. All but one began use of anti-impaction gloves in the workplace. RESULTS: All patients with FHD were able to return to work with the use of anti-impaction gloves, topical treatments, and (in some cases) modified work duties. Neither of the two cases of HHD resolved with treatment or with discontinuation of work. CONCLUSION: Anti-impaction gloves are shown to aid in the return-to-work process for those with FHD but not for those with HHD. PMID- 19808005 TI - Questionnaire study of the prevalence of allergic contact dermatitis from cosmetics in Israel. AB - BACKGROUND: The prevalence of cosmetics-related contact dermatitis is rising, owing mainly to the wider use of cosmetics by the public and the routine diagnostic use of patch tests. OBJECTIVE: To determine the prevalence of cosmetics allergy in Israel. METHODS: A questionnaire was distributed to a random sample of 360 female customers of pharmacies and beauty salons in two areas of the country. Items included general health profile, family history of atopy, occurrence of rash due to patch-test-proven cosmetics allergy, anatomic sites of the rash, subjective aspects regarding the rash, and cosmetics consumption habits. RESULTS: Patient age ranged from 15 to 89 years. Eleven subjects (3.1%) had patch-test-proven cosmetics allergic contact dermatitis. There was a correlation between proven cosmetics allergy and subjective sensitivity to facial cream (p = .03). CONCLUSIONS: The 3.1% prevalence rate of cosmetics contact allergic dermatitis in a randomly selected population in Israel is similar to values reported in the literature (about 2%). The higher-than-expected rate of subjective sensitivity to facial cream among patients with proven cosmetics allergy may be explained by the wide use of facial cream, facial skin susceptibility to insult, and the relatively long duration of contact of facial cream (a leave-on product) with the skin. PMID- 19808006 TI - Contact sensitization by age group in adults: patch-test data from the Massachusetts General Hospital, 1996 to 2006. AB - BACKGROUND: Allergic contact sensitization may occur at any age, showing specific rates of sensitization and allergen frequency related to periods of life. OBJECTIVE: To describe sensitization rates of specific allergens with respect to age distribution in adults in a cross-sectional study. METHODS: Data of 713 patients referred for patch testing at the Massachusetts General Hospital Contact Dermatitis Clinic were analyzed. Testing was conducted with baseline (29 allergens) and customized series between January 1996 and December 2006. RESULTS: Although trends were noted, there was no statistically significant difference with respect to rate of sensitization or number of positive allergens in the various age groups. Fragrance mix and nickel sulfate were the most common allergens through all age groups. In older age groups, a statistically significant higher sensitization rate was shown for fragrance mix (p = .005) and balsam of Peru (p < .005) whereas the rate for nickel was lower (p = .002). CONCLUSIONS: For specific allergens such as nickel and fragrances, significant variations occur in different stages of life. Even with these variations, we found that the overall rates of positive patch-test reactions were similar in the three age groups evaluated. PMID- 19808007 TI - Allergic contact dermatitis from carmine. AB - A 28-year-old woman developed allergic contact dermatitis within 6 to 24 hours exclusively after using carmine-containing eyeshadows and lipsticks. She had both a positive patch test result and a positive antecubital repeated open application test result with carmine 2.5% in petrolatum. Thirty other patients had negative patch test results. Carmine is a widely used pigment derived from gravid cochineal insects. Carminic acid is the source of its color. Only two previous publications describing allergic contact dermatitis from carmine could be found. The ingredient in carmine causing these delayed hypersensitivity reactions has not been studied. In contrast, there are numerous reports of immediate hypersensitivity reactions from carmine, mostly from its use in foods and beverages but also from cosmetics and pharmaceuticals. These are immunoglobulin E mediated reactions directed against cochineal proteins. PMID- 19808008 TI - Facial flushing: an uncommon presentation of serotonin toxicity. PMID- 19808009 TI - Allergic contact dermatitis from tricresyl phosphate. PMID- 19808013 TI - Sphingosine-1-phosphate and oligodendrocytes: from cell development to the treatment of multiple sclerosis. AB - Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid that mediates a wide variety of biological effects in different cells and tissues. This review discusses the effects of S1P signaling in oligodendrocytes, the myelin making cells of the central nervous system (CNS). Results from different laboratories have uncovered direct actions of S1P at different maturational stages along the oligodendroglial lineage. There is also evidence for the existence in oligodendrocytes of interactions between S1P and signaling by factors which, like neurotrophin-3 (NT-3) and platelet-derived growth factor (PDGF), have profound effects on oligodendrocyte development and myelination. Moreover, S1P signaling in oligodendrocytes may not only play an important role during normal CNS development but also offer new therapeutic avenues to stimulate remyelination in demyelinating diseases like multiple sclerosis. PMID- 19808014 TI - Does hypothalamic inflammation cause obesity? AB - Obesity-induced inflammation causes cellular resistance to both insulin and leptin. In this issue, Bruning and colleagues (Kleinridders et al., 2009) add to growing evidence that this response occurs in the hypothalamus, as well as in peripheral tissues, which helps to explain how high-fat feeding induces a gradual increase in defended body weight. PMID- 19808012 TI - Emerging roles of PGE2 receptors in models of neurological disease. AB - This review presents an overview of the emerging field of prostaglandin signaling in neurological diseases, focusing on PGE(2) signaling through its four E prostanoid (EP) receptors. A large number of studies have demonstrated a neurotoxic function of the inducible cyclooxygenase COX-2 in a broad spectrum of neurological disease models in the central nervous system (CNS), from models of cerebral ischemia to models of neurodegeneration and inflammation. Since COX-1 and COX-2 catalyze the first committed step in prostaglandin synthesis, an effort is underway to identify the downstream prostaglandin signaling pathways that mediate the toxic effect of COX-2. Recent epidemiologic studies demonstrate that chronic COX-2 inhibition can produce adverse cerebrovascular and cardiovascular effects, indicating that some prostaglandin signaling pathways are beneficial. Consistent with this concept, recent studies demonstrate that in the CNS, specific prostaglandin receptor signaling pathways mediate toxic effects in brain but a larger number appear to mediate paradoxically protective effects. Further complexity is emerging, as exemplified by the PGE(2) EP2 receptor, where cerebroprotective or toxic effects of a particular prostaglandin signaling pathway can differ depending on the context of cerebral injury, for example, in excitotoxicity/hypoxia paradigms versus inflammatory-mediated secondary neurotoxicity. The divergent effects of prostaglandin receptor signaling will likely depend on distinct patterns and dynamics of receptor expression in neurons, endothelial cells, and glia and the specific ways in which these cell types participate in particular models of neurological injury. PMID- 19808015 TI - Leptin's RIGHT turn to the brain stem. AB - In a paper by Yadav and colleagues, a novel pathway linking the central nervous system effects of leptin on bone mass and energy expenditure to serotonin signaling in brainstem circuits is described. The data from those studies strengthen the tenet that skeletal remodeling is intimately connected to central regulation of metabolism. PMID- 19808016 TI - A new wrinkle in the fold: hepcidin links inflammation to the unfolded protein response. AB - The unfolded protein response (UPR) coordinates translational and transcriptional changes triggered by unfolded proteins within the endoplasmic reticulum. Two recent papers (Oliveira et al., 2009; Vecchi et al., 2009) show that the UPR modulates transcription of the hormone hepcidin, which controls plasma iron levels and perhaps innate immunity. PMID- 19808017 TI - Translate this ... during dietary restriction. AB - Several studies indicate that reduced TOR signaling underlies life span extension by dietary restriction. Recently, Zid et al. (Zid et al., 2009) linked the benefits of dietary restriction in flies to increased levels of the downstream TOR target 4E-BP1 and corresponding changes in the relative translation rates of classes of mRNAs. PMID- 19808018 TI - MyD88 signaling in the CNS is required for development of fatty acid-induced leptin resistance and diet-induced obesity. AB - Obesity-associated activation of inflammatory pathways represents a key step in the development of insulin resistance in peripheral organs, partially via activation of TLR4 signaling by fatty acids. Here, we demonstrate that palmitate acting in the central nervous system (CNS) inhibits leptin-induced anorexia and Stat3 activation. To determine the functional significance of TLR signaling in the CNS in the development of leptin resistance and diet-induced obesity in vivo, we have characterized mice deficient for the TLR adaptor molecule MyD88 in the CNS (MyD88(DeltaCNS)). Compared to control mice, MyD88(DeltaCNS) mice are protected from high-fat diet (HFD)-induced weight gain, from the development of HFD-induced leptin resistance, and from the induction of leptin resistance by acute central application of palmitate. Moreover, CNS-restricted MyD88 deletion protects from HFD- and icv palmitate-induced impairment of peripheral glucose metabolism. Thus, we define neuronal MyD88-dependent signaling as a key regulator of diet-induced leptin and insulin resistance in vivo. PMID- 19808019 TI - Reactive oxygen species enhance insulin sensitivity. AB - Chronic reactive oxygen species (ROS) production by mitochondria may contribute to the development of insulin resistance, a primary feature of type 2 diabetes. In recent years it has become apparent that ROS generation in response to physiological stimuli such as insulin may also facilitate signaling by reversibly oxidizing and inhibiting protein tyrosine phosphatases (PTPs). Here we report that mice lacking one of the key enzymes involved in the elimination of physiological ROS, glutathione peroxidase 1 (Gpx1), were protected from high-fat diet-induced insulin resistance. The increased insulin sensitivity in Gpx1(-/-) mice was attributed to insulin-induced phosphatidylinositol-3-kinase/Akt signaling and glucose uptake in muscle and could be reversed by the antioxidant N acetylcysteine. Increased insulin signaling correlated with enhanced oxidation of the PTP family member PTEN, which terminates signals generated by phosphatidylinositol-3-kinase. These studies provide causal evidence for the enhancement of insulin signaling by ROS in vivo. PMID- 19808020 TI - MicroRNA-210 controls mitochondrial metabolism during hypoxia by repressing the iron-sulfur cluster assembly proteins ISCU1/2. AB - Repression of mitochondrial respiration represents an evolutionarily ancient cellular adaptation to hypoxia and profoundly influences cell survival and function; however, the underlying molecular mechanisms are incompletely understood. Primarily utilizing pulmonary arterial endothelial cells as a representative hypoxic cell type, we identify the iron-sulfur cluster assembly proteins (ISCU1/2) as direct targets for repression by the hypoxia-induced microRNA-210 (miR-210). ISCU1/2 facilitate the assembly of iron-sulfur clusters, prosthetic groups that are critical for electron transport and mitochondrial oxidation-reduction reactions. Under in vivo conditions of upregulating miR-210 and repressing ISCU1/2, the integrity of iron-sulfur clusters is disrupted. In turn, by repressing ISCU1/2 during hypoxia, miR-210 decreases the activity of prototypical iron-sulfur proteins controlling mitochondrial metabolism, including Complex I and aconitase. Consequently, miR-210 represses mitochondrial respiration and associated downstream functions. These results identify important mechanistic connections among microRNA, iron-sulfur cluster biology, hypoxia, and mitochondrial function, with broad implications for cellular metabolism and adaptation to cellular stress. PMID- 19808021 TI - Loss of Lkb1 in adult beta cells increases beta cell mass and enhances glucose tolerance in mice. AB - The Lkb1 tumor suppressor exerts its biological effects through phosphorylation and consequent activation of the AMP kinase (AMPK) family. Extensive genetic and biochemical evidence supports a role for Lkb1 in cell cycle arrest, establishment of cell polarity, and cellular energy metabolism. However, the role of Lkb1 and the AMPK family in beta cell function in vivo has not been established. We generated conditional knockout mice with a deletion of the Lkb1 gene in the beta cell compartment of pancreatic islets; these mice display improved glucose tolerance and protection against diet-induced hyperglycemia. Lkb1(-/-) beta cells are hypertrophic because of elevated mTOR activity; they also proliferate more and secrete more insulin in response to glucose. These data indicate that inhibiting Lkb1 activity in beta cells may facilitate beta cell expansion and glucose tolerance in vivo. PMID- 19808022 TI - LKB1 regulates pancreatic beta cell size, polarity, and function. AB - Pancreatic beta cells, organized in the islets of Langerhans, sense glucose and secrete appropriate amounts of insulin. We have studied the roles of LKB1, a conserved kinase implicated in the control of cell polarity and energy metabolism, in adult beta cells. LKB1-deficient beta cells show a dramatic increase in insulin secretion in vivo. Histologically, LKB1-deficient beta cells have striking alterations in the localization of the nucleus and cilia relative to blood vessels, suggesting a shift from hepatocyte-like to columnar polarity. Additionally, LKB1 deficiency causes a 65% increase in beta cell volume. We show that distinct targets of LKB1 mediate these effects. LKB1 controls beta cell size, but not polarity, via the mTOR pathway. Conversely, the precise position of the beta cell nucleus, but not cell size, is controlled by the LKB1 target Par1b. Insulin secretion and content are restricted by LKB1, at least in part, via AMPK. These results expose a molecular mechanism, orchestrated by LKB1, for the coordinated maintenance of beta cell size, form, and function. PMID- 19808023 TI - Suppression of sulfonylurea- and glucose-induced insulin secretion in vitro and in vivo in mice lacking the chloride transport protein ClC-3. AB - Priming of insulin secretory granules for release requires intragranular acidification and depends on vesicular Cl(-)-fluxes, but the identity of the chloride transporter/ion channel involved is unknown. We tested the hypothesis that the chloride transport protein ClC-3 fulfills these actions in pancreatic beta cells. In ClC-3(-/-) mice, insulin secretion evoked by membrane depolarization (high extracellular K(+), sulfonylureas), or glucose was >60% reduced compared to WT animals. This effect was mirrored by a approximately 80% reduction in depolarization-evoked beta cell exocytosis (monitored as increases in cell capacitance) in single ClC-3(-/-) beta cells, as well as a 44% reduction in proton transport across the granule membrane. ClC-3 expression in the insulin granule was demonstrated by immunoblotting, immunostaining, and negative immuno EM in a high-purification fraction of large dense-core vesicles (LDCVs) obtained by phogrin-EGFP labeling. The data establish the importance of granular Cl(-) fluxes in granule priming and provide direct evidence for the involvement of ClC 3 in the process. PMID- 19808024 TI - The granular chloride channel ClC-3 is permissive for insulin secretion. AB - Insulin secretion from pancreatic beta cells is dependent on maturation and acidification of the secretory granule, processes necessary for prohormone convertase cleavage of proinsulin. Previous studies in isolated beta cells revealed that acidification may be dependent on the granule membrane chloride channel ClC-3, in a step permissive for a regulated secretory response. In this study, immuno-EM of beta cells revealed colocalization of ClC-3 and insulin on secretory granules. Clcn3(-/-) mice as well as isolated islets demonstrate impaired insulin secretion; Clcn3(-/-) beta cells are defective in regulated insulin exocytosis and granular acidification. Increased amounts of proinsulin were found in the majority of secretory granules in the Clcn3(-/-) mice, while in Clcn3(+/+) cells, proinsulin was confined to the immature secretory granules. These results demonstrate that in pancreatic beta cells, chloride channels, specifically ClC-3, are localized on insulin granules and play a role in insulin processing as well as insulin secretion through regulation of granular acidification. PMID- 19808025 TI - Proteome differences between brown and white fat mitochondria reveal specialized metabolic functions. AB - Mitochondria are functionally specialized in different tissues, and a detailed understanding of this specialization is important to elucidate mitochondrial involvement in normal physiology and disease. In adaptive thermogenesis, brown fat converts mitochondrial energy to heat, whereas tissue-specific functions of mitochondria in white fat are less characterized. Here we apply high-resolution quantitative mass spectrometry to directly and accurately compare the in vivo mouse mitochondrial proteomes of brown and white adipocytes. Their proteomes are substantially different qualitatively and quantitatively and are furthermore characterized by tissue-specific protein isoforms, which are modulated by cold exposure. At transcript and proteome levels, brown fat mitochondria are more similar to their counterparts in muscle. Conversely, white fat mitochondria not only selectively express proteins that support anabolic functions but also degrade xenobiotics, revealing a protective function of this tissue. In vivo comparison of organellar proteomes can thus directly address functional questions in metabolism. PMID- 19808026 TI - Identification of a synthetic peptide inducing cross-reactive antibodies binding to Rhipicephalus (Boophilus) decoloratus, Rhipicephalus (Boophilus) microplus, Hyalomma anatolicum anatolicum and Rhipicephalus appendiculatus BM86 homologues. AB - The BM86 antigen, originally identified in Rhipicephalus (Boophilus) microplus, is the basis of the only commercialized anti-tick vaccine. The long-term goal of our study is to improve BM86 based vaccines by induction of high levels of tick gut binding antibodies that are also cross-reactive with a range of BM86 homologues expressed in other important tick species. Here we have used a BD86 derived synthetic peptide, BD86-3, to raise a series of mouse monoclonal antibodies. One of these mAbs, named 12.1, recognized BM86 homologues in immuno histochemical analyses in four out of five tick species including R. (B.) microplus, Rhipicephalus (Boophilus) decoloratus, Hyalomma anatolicum anatolicum and Rhipicephalus appendiculatus. Our results indicate that broadly cross reactive tick gut binding antibodies can be induced after immunization with a synthetic peptide derived from the protein BD86. PMID- 19808027 TI - Small fiber neuropathy following vaccination for rabies, varicella or Lyme disease. AB - Neuropathy following vaccination has been reported; however, biopsy-confirmed small fiber neuropathy has not been described. We report five patients who developed paresthesias within one day to two months following vaccination for rabies, varicella zoster, or Lyme disease. On examination, there was mild sensory loss in distal extremities, preserved strength, normal or minimally abnormal electrodiagnostic findings, and decreased epidermal nerve fiber densities per skin biopsy. Empiric immunomodulatory therapy was tried in two patients and was ineffective. All patients' symptoms have improved, but persist. We conclude that an acute or subacute, post-vaccination small fiber neuropathy may occur and follow a chronic course. PMID- 19808028 TI - Recombinant pro-apoptotic Mycobacterium tuberculosis generates CD8+ T cell responses against human immunodeficiency virus type 1 Env and M. tuberculosis in neonatal mice. AB - Mycobacterium bovis BCG is an attractive vaccine vector against breast milk HIV transmission because it elicits Th1-type responses in newborns. However, BCG causes disease in HIV-infected infants. Genetically attenuated Mycobacterium tuberculosis (Mtb) mutants represent a safer alternative for immunocompromised populations. In the current study, we compared the immunogenicity in mice of three different recombinant attenuated Mtb strains expressing an HIV envelope (Env) antigen construct. Two of these strains (DeltalysA DeltapanCD Mtb and DeltaRD1 DeltapanCD Mtb) failed to induce significant levels of HIV Env-specific CD8(+) T cell responses. In striking contrast, an HIV-1 Env-expressing attenuated DeltalysA Mtb containing a deletion in secA2, which encodes a virulence-related secretion system involved in evading adaptive immunity, generated consistently measurable Env-specific CD8(+) T cell responses that were significantly greater than those observed after immunization with BCG expressing HIV Env. Similarly, another strain of DeltalysA DeltasecA2 Mtb expressing SIV Gag induced Gag- and Mtb-specific CD8(+) T cells producing perforin or IFNgamma, and Gag-specific CD4(+) T cells producing IFNgamma within 3 weeks after immunization in adult mice; in addition, IFNgamma-producing Gag-specific CD8(+) T cells and Mtb specific CD4(+) T cells were observed in neonatal mice within 1 week of immunization. We conclude that DeltalysA DeltasecA2 Mtb is a promising vaccine platform to construct a safe combination HIV-TB vaccine for use in neonates. PMID- 19808029 TI - Preclinical and clinical development of YFV 17D-based chimeric vaccines against dengue, West Nile and Japanese encephalitis viruses. AB - Dengue viruses (DENV), West Nile virus (WNV) and Japanese encephalitis virus (JEV) are major global health and growing medical problems. While a live attenuated vaccine exists since decades against the prototype flavivirus, yellow fever virus (YFV), there is an urgent need for vaccines against dengue or West Nile diseases, and for improved vaccines against Japanese encephalitis. Live attenuated chimeric viruses were constructed by replacing the genes coding for Premembrane (prM) and Envelope (E) proteins from YFV 17D vaccine strain with those of heterologous flaviviruses (ChimeriVax technology). This technology has been used to produce vaccine candidates for humans, for construction of a horse vaccine for West Nile fever, and as diagnostic reagents for dengue, Japanese encephalitis, West Nile and St. Louis encephalitis infections. This review focuses on human vaccines and their characterization from the early stages of research through to clinical development. Phenotypic and genetic properties and stability were examined, preclinical evaluation through in vitro or animal models, and clinical testing were carried out. Theoretical environmental concerns linked to the live and genetically modified nature of these vaccines have been carefully addressed. Results of the extensive characterizations are in accordance with the immunogenicity and excellent safety profile of the ChimeriVax-based vaccine candidates, and support their development towards large-scale efficacy trials and registration. PMID- 19808030 TI - Plasma chitotriosidase and CCL18 as surrogate markers for granulomatous macrophages in sarcoidosis. AB - BACKGROUND: Accumulation of macrophages in multiple organs is a common feature of sarcoidosis and Gaucher disease. The vast number of storage macrophages in Gaucher patients has facilitated the discovery of suitable plasma markers like chitotriosidase and CCL18. METHODS: Plasma specimens of patients with sarcoidosis were examined on chitotriosidase activity and CCL18 protein levels. RESULTS: Chitotriosidase was markedly increased, being on average 13.7-fold elevated (range: 1.1-43.3). The sensitivity of demonstrating sarcoidosis using plasma chitotriosidase values exceeded that using serum angiotensin-converting enzyme values. A 3.5-fold (range: 1-15) increase in CCL18 was also observed. The relative changes in chitotriosidase and CCL18 during the course of disease closely mimicked each other, suggesting an identical cellular source. In situ hybridization analysis confirmed massive production of chitotriosidase by sarcoid macrophages. The increase in plasma chitotriosidase correlated with the stage of disease, being highest in active sarcoidosis with extrapulmonary involvement. Therapy with steroids resulted in clear reduction of plasma chitotriosidase and CCL18 and relapse of disease activity was preceded by increases in these parameters. CONCLUSIONS: Sarcoid macrophages secrete high quantities of chitotriosidase and CCL18. Determination of plasma chitotriosidase and CCL18 may be useful to monitor changes in granulomatous macrophages during the course of sarcoidosis. PMID- 19808031 TI - ESI-MS/MS quantification of 7alpha-hydroxy-4-cholesten-3-one facilitates rapid, convenient diagnostic testing for cerebrotendinous xanthomatosis. AB - BACKGROUND: The genetic disorder cerebrotendinous xanthomatosis (CTX) frequently remains undiagnosed for many years. Left untreated CTX is associated with the development of cataracts, xanthomas and severe neurological dysfunction. The method routinely used to screen for CTX is GC-based measurement of elevated 5alpha-cholestanol from hydrolyzed plasma. A plasma test for CTX utilizing ESI MS/MS methodology would be beneficial. METHODS: Development of rapid, simple LC ESI-MS/MS methodology to test plasma for CTX is described. Two hour Girard derivatization allowed for 7alpha-hydroxy-4-cholesten-3-one quantification by isotope dilution LC-ESI-MS/MS within 12 min from un-hydrolyzed affected patient plasma (5 microl). RESULTS: Adequate sensitivity and reproducibility were achieved for quantification of 7alpha-hydroxy-4-cholesten-3-one, which demonstrated improved utility as a diagnostic marker of disease and to monitor treatment compared to 5alpha-cholestanol. The mean plasma concentration of 7alpha hydroxy-4-cholesten-3-one in untreated CTX-affected patients (n=6) was 107-fold that in unaffected subjects (n=9), with the lowest concentration in affected patients >10-fold the highest concentration in unaffected subjects. CONCLUSION: Quantification of the bile acid precursor 7alpha-hydroxy-4-cholesten-3-one with LC-ESI-MS/MS is a novel approach to improved diagnostic testing of plasma for CTX, amenable to high-throughput analysis and automated sample handling. Development of ESI-MS/MS methodology should make CTX testing more widely available and facilitate easier diagnosis of CTX. PMID- 19808032 TI - Predictive value of thrombopath determination in women with infertility and pregnancy complications. AB - BACKGROUND: A condition of maternal thrombophilia, either inherited or acquired, can compromise the success of implantation and foetal survival. METHODS: Malfunctions in protein C pathway were evaluated using a novel assay [HemosIL ThromboPath (ThP)] in a case-control study of 172 women with a history of recurrent miscarriage or infertility and 86 age-matched unrelated fertile women. RESULTS: Thrombophilia was ascertained in 13% of the cases. ThP values were lower in women either with or without thrombophilia compared to controls (both p<0.0001). Abnormal ThP values (below the mean minus 1SD of controls) were found in 62% of cases compared to 12% of controls (p<0.0001). Non-thrombophilic women who achieved spontaneous pregnancy had higher ThP values compared to those who did not (p<0.05). Elevated ThP values were an independent predictor for pregnancy (p<0.01) in women without thrombophilia. A decrease of ThP values was observed during pregnancy progression, which correlated with that of protein S (p<0.05). Miscarriage or major complications occurred in women in whom ThP percent change was approximately 2-fold higher than that observed in women who achieved successful pregnancy (p<0.05). CONCLUSIONS: ThP might represent an efficient assay for screening women with pregnancy complications and might provide prognostic information during pregnancy progression. PMID- 19808033 TI - The role of MSH5 C85T and MLH3 C2531T polymorphisms in the risk of male infertility with azoospermia or severe oligozoospermia. AB - BACKGROUND: The mismatch repair proteins MSH5 and MLH3 play a crucial role in spermatogenesis. We tested this hypothesis by examining the contribution of functional polymorphisms in MSH5 C85T and MLH3 C2531T to the risk of male infertility. METHODS: We investigated Chinese patients, including 162 infertile individuals with idiopathic azoospermia or severe oligozoospermia, and 160 fertile men as controls. RESULTS: We observed an increased risk of male infertility associated with the MSH5 (CT+TT) (OR, 2.51; 95% CI, 1.43-4.40; P<0.001) or MLH3 (CT+TT) (OR, 1.98; 95% CI, 1.23-3.17; P<0.001) genotype, compared to the MSH5 CC or MLH3 CC genotype, respectively. Interactions between these MSH5 and MLH3 polymorphisms increased the risk of male infertility in a multiplicative manner, with the OR being 6.78 (95% CI, 2.12-21.68) for subjects carrying both MSH5 (CT+TT) and MLH3 (CT+TT) genotypes. CONCLUSIONS: There is an association of polymorphism C85T in MSH5 or C2531T in MLH3 with male infertility, specifically azoospermia or severe oligozoospermia, and interaction between these MSH5 and MLH3 polymorphisms increased the risk of developing male infertility. Therefore, the MSH5 and MLH3 polymorphisms may be genetic determinants for human spermatogenesis impairment. PMID- 19808034 TI - Recombinant plant gamma carbonic anhydrase homotrimers bind inorganic carbon. AB - Gamma carbonic anhydrases (gammaCA) are widespread in Prokaryotes. In Eukaryotes, homologous genes were found only in plant genomes. In Arabidopsis and maize, the corresponding gene products are subunits of mitochondrial Complex I. At present, only gammaCA homotrimers of Methanosarcina thermophila (CAM) show reversible carbon dioxide (CO(2)) hydration activity. In the present work, it is shown that recombinant plant gammaCA2 could form homotrimers and bind H(14)CO(3)(-). However, they are unable to catalyse the reversible hydration of CO(2). These results suggest that plant gammaCAs do not act as carbonic anhydrases but with a related activity possibly contributing to recycle CO(2) in the context of photorespiration. PMID- 19808035 TI - Cx40.8, a Cx43-like protein, forms gap junction channels inefficiently and may require Cx43 for its association at the plasma membrane. AB - In addition to having a Cx43 ortholog, the zebrafish genome also contains a Cx43 like gene, Cx40.8. Here, we investigate the expression of cx40.8 in zebrafish fins and the function of Cx40.8 in HeLa cells. We find that cx40.8 is present in the same population of dividing cells as cx43. Unlike Cx43, dye coupling assays suggest that Cx40.8 only inefficiently forms functional gap junction channels. However, co-transfection reveals that Cx40.8 can co-localize with Cx43 in gap junction plaques, and that the resulting plaques contain functional gap junction channels. Together, these data suggest the possibility that Cx40.8 may functionally interact with Cx43 to regulate cell proliferation in vivo. PMID- 19808036 TI - Plant growth promotion due to rhizobacterial volatiles--an effect of CO2? AB - Serratia odorifera, an antagonistic rhizobacterium, emits a diverse and complex bouquet of volatiles. Three different in vitro experimental culture systems indicated that these volatiles promote the growth of Arabidopsis thaliana. CO(2) trapping and significant rise of CO(2) levels (390-3000 ppm CO(2) within 24 h) due to bacterial growth in sealed Petri dishes verified the enhanced effects of rhizobacterial CO(2) on A. thaliana's growth. In contrast, open cocultivations abrogated growth promotion, and inhibitory effects come to the fore at ambient CO(2) concentrations. PMID- 19808037 TI - In vitro effects on bacterial growth of phenoloxidase reaction products. AB - An active phenoloxidase preparation from the freshwater crayfish Pacifastacus leniusculus exhibited a strong antibacterial effect in vitro on the bacteria Aeromonas hydrophila, Escherichia coli, Streptococcus pneumoniae whereas a weaker but still significant effect against Bacillus cereus, Pseudomonas aeruginosa and Staphylococcus aureus. In most cases reduction of bacterial growth was stronger when dopamine was used as substrate as compared to L-dopa. The effect on bacteria was abolished if no substrate was available for the phenoloxidase or in the presence of the phenoloxidase inhibitor phenylthiourea. PMID- 19808038 TI - Costly dispersal can destabilize the homogeneous equilibrium of a metapopulation. AB - I investigate the stability of the homogeneous equilibrium of a discrete-time metapopulation assuming costly dispersal with arbitrary (but fixed) spatial pattern of connectivity between the local populations. First, I link the stability of the metapopulation to the stability of a single isolated population by proving that the homogeneous metapopulation equilibrium, provided that it exists, is stable if and only if a single population, which is subject to extra mortality matching the average dispersal-induced mortality of the metapopulation, has a stable fixed point. Second, I demonstrate that extra mortality may destabilize the fixed point of a single population. Taken together, the two results imply that costly dispersal can destabilize the homogeneous equilibrium of a metapopulation. I illustrate this by simulations and discuss why earlier work, arriving at the opposite conclusion, was flawed. PMID- 19808039 TI - Several appropriate background distributions for entropy-based protein sequence conservation measures. AB - Amino acid background distribution is an important factor for entropy-based methods which extract sequence conservation information from protein multiple sequence alignments (MSAs). However, MSAs are usually not large enough to allow a reliable observed background distribution. In this paper, we propose two new estimations of background distribution. One is an integration of the observed background distribution and the position-specific residue distribution, and the other is a normalized square root of observed background frequency. To validate these new background distributions, they are applied to the relative entropy model to find catalytic sites and ligand binding sites from protein MSAs. Experimental results show that they are superior to the observed background distribution in predicting functionally important residues. PMID- 19808040 TI - Sex-biased dispersal of adults mediates the evolution of altruism among juveniles. AB - Population viscosity has been proposed as an important mechanism for the evolution of cooperation. The idea is that if individuals do not disperse far during the course of their lives, they will tend to interact with their genealogical relatives, which may give kin-selected benefits for cooperation. However, in the simplest model of population structure, the evolution of cooperation is unaffected by the rate of dispersal, owing to dispersal also mediating competition between social partners. This surprising result has generated much research interest in recent years. Here I show that dispersal does matter if there is a sex difference in dispersal rate, even when the expression of cooperation is not conditional upon the actor's dispersal status or sex. In particular, I show that cooperation among juveniles is relatively favoured when there is a small sex bias in adult dispersal in favour of the sex with the greatest variance in reproductive success, and is relatively disfavoured when this sex bias is large or in the opposite direction. This is because dispersal by individuals of each sex can have different consequences for the genetic structure of the population. PMID- 19808041 TI - Hybrid cellular automaton modeling of nutrient modulated cell growth in tissue engineering constructs. AB - Mathematic models help interpret experimental results and accelerate tissue engineering developments. We develop in this paper a hybrid cellular automata model that combines the differential nutrient transport equation to investigate the nutrient limited cell construct development for cartilage tissue engineering. Individual cell behaviors of migration, contact inhibition and cell collision, coupled with the cell proliferation regulated by oxygen concentration were carefully studied. Simplified two-dimensional simulations were performed. Using this model, we investigated the influence of cell migration speed on the overall cell growth within in vitro cell scaffolds. It was found that intense cell motility can enhance initial cell growth rates. However, since cell growth is also significantly modulated by the nutrient contents, intense cell motility with conventional uniform cell seeding method may lead to declined cell growth in the final time because concentrated cell population has been growing around the scaffold periphery to block the nutrient transport from outside culture media. Therefore, homogeneous cell seeding may not be a good way of gaining large and uniform cell densities for the final results. We then compared cell growth in scaffolds with various seeding modes, and proposed a seeding mode with cells initially residing in the middle area of the scaffold that may efficiently reduce the nutrient blockage and result in a better cell amount and uniform cell distribution for tissue engineering construct developments. PMID- 19808042 TI - A mechanistic model of nutritional control of protein synthesis in animal tissues. AB - Regulation of mRNA translation has been held responsible for effects of diet, age, alcohol, hormones, hibernation, disease and hypoxia on protein synthesis in animal tissues. Dietary effects are due to concentrations of amino acids and insulin in circulation that affect activities of two key translational regulators, eukaryotic initiation factor 2 (F2) and eukaryotic initiation factor 4E binding protein 1 (Bp). To construct a platform for prediction of global protein synthesis to nutritional stimuli, a dynamic, mechanistic model of translational control in whole tissues was developed. The model was composed of a set of differential equations which describe the dynamics of 11 state variables: tRNA and acyl-tRNA for leucine (Leu), limiting (Laa) and other amino acids (Oaa), inactivated F2 with GDP (F2d), activated F2 with GTP (F2t), F4e, Bp and its complex with F4e (4eBp), available mRNA start codons (AUG), and active ribosomes (Arib). Material was assumed to flow from one variable to another according to mass-action kinetics or Michaelis-Menten form. Uncharged tRNA inhibit GTP exchange on eIF2, and free amino acids and insulin inhibit reversible sequestration of F4e by Bp. Initial conditions and parameters were set for a skeletal muscle fractional synthesis rate of 10%/d and ribosome transit time of 80s. Between amino acid concentrations of 500 and 4000x10(3)nM, protein synthesis increased from 0.9 to 11.7%/d at 0microU/mL insulin, and from 5.0 to 12.8%/d at 30microU/mL insulin. Predicted responses to graded levels of a deficient amino acid were asymptotic. A single parameter accomodated differences between tissues in insulin sensitivity. Seven parameters must be changed to simulate initiation and elongation rates in more active tissues such as liver, or in tissues of older mature animals. An increase in uncharged tRNA during insulin stimulation highlighted the physiological importance of coordinated regulation of amino acid supply by insulin. In conclusion, the regulation of F4e release from Bp by Ins and Leu, and of F2d recycling by uncharged tRNA can be tied together to describe a wide range of FSR values across tissues and physiological states. PMID- 19808043 TI - Aging reduces reproductive success in mussels Mytilus edulis. AB - The present study was aimed to determine whether reproductive success constantly increases with age in a relatively short-lived invertebrate with continuous growth - the bivalve mollusc Mytilus edulis or there is an age-related decline such as observed in species with finite growth (mammals, insects, nematodes, etc.). We studied the reproductive output and viability of the offspring during early embryogenesis in females of different sizes and ages, and used allometric approaches to correct for the effects of the body size and to discern pure age specific effects on these reproductive traits. We have also determined contributions of females of different age and size classes to the total larval pool of a population. Both gonadosomatic index and individual fecundity significantly decreased in the course of aging if the size of the animals was accounted for. The proportion of normally developing embryos declined from almost 100% to 60% in females of 2-10-year-old. We suggest that animals with infinite growth and "slow aging", such as molluscs, undergo senescence, the physiological manifestations of which can be masked by a more pronounced effect of continuously increasing size. PMID- 19808044 TI - Melatonin alleviates memory deficits and neuronal degeneration induced by intracerebroventricular administration of streptozotocin in rats. AB - In the present study the effect of melatonin on intracerebroventricularly administered streptozotocin (STZ)-induced neurodegeneration was investigated in rats. STZ (3mg/kg), administered twice with an interval of 48 h between the two doses, showed impairment in spatial memory tested by water maze test after 14 days of 1st dose. Administration of melatonin (2.5, 5.0 and 10mg/kg, i.p.) was started 1h prior to 1st dose of STZ and continued up to 14 days. Glutathione and malondialdehyde were used as biochemical markers of oxidative stress in different brain regions. Histopathological changes were examined by using hematoxylin and eosin stain. STZ administration caused significant decrease in glutathione and increase in malondialdehyde as compared to control and artificial Cerebrospinal Fluid treated rats indicating oxidative stress. Brain sections of STZ-treated rats showed increased vacuoles in the periventricular cortical area, damaged periventricular cells and damaged cells in the hippocampal CA4 region as compared to control and artificial Cerebrospinal Fluid treated groups. Melatonin treatment significantly attenuated the effect of STZ-induced oxidative stress and histopathological changes. The results indicate that melatonin is effective in providing protection against memory deficit, oxidative stress and neuronal damage induced by STZ. PMID- 19808045 TI - Emerging role of cyclooxygenase isoforms in the control of gastrointestinal neuromuscular functions. AB - Since the discovery of two cyclooxygenase isoforms (COX-1, COX-2), efforts have been made to characterize the roles played by these enzymes in the regulation of physiological functions, as well as to explore their involvement in the pathophysiology of inflammatory disorders. In the digestive tract, the majority of evidence has been obtained at mucosal level, where both isoforms regulate various functions, and contribute to the development of inflammatory and neoplastic disorders. The role of COX isoforms in the gut neuromuscular compartment, where their expression has been detected in different species, is still unclear. However, the characterization of actions exerted by COX-derived prostanoids on gut motility has been under investigation for many years, and it is becoming increasingly appreciated that these mediators subserve complex regulatory patterns of COX on digestive motility. More recently, several studies have strengthened the concept that both COX-1 and COX-2 are involved in the modulation of gastrointestinal neuromuscular activity under normal conditions, and that changes in their regulatory activities occur in the presence of various digestive disorders, including inflammatory bowel diseases and postoperative ileus. Despite a large body of preclinical evidence, studies aimed at translating these findings into clinically relevant applications are needed, in an attempt to identify novel therapeutic approaches for treatment of gut disorders associated with motility alterations. This review illustrates and discusses current knowledge of the roles played by COX pathways in the regulation of gastrointestinal neuromuscular functions, both under normal conditions and in the presence of gut disorders. PMID- 19808046 TI - B cell activation versus anergy; the antigen receptor as a molecular switch. AB - Anergic B cells are autoreactive and are present in the periphery in an unresponsive state. Here we will discuss the difference in B cell receptor signaling between anergic B cells, chronically stimulated by autoantigen, and naive B cell encountering antigen. PMID- 19808047 TI - Unraveling bacterial interactions with toll-like receptors. AB - Bacterial diversity, tight regulation of Toll-like receptor function, and the variation of cellular phenotypes dependent on the local microenvironment, are increasingly recognized as key factors in maintaining immune homeostasis. Here we discuss bacterial and host factors important for a balanced immune response during infection. PMID- 19808048 TI - Fc receptors: role in biology and antibody therapy. AB - Insight into the biological role of antibody (Fc) receptors on immune and endothelial cells increased profoundly over the last decades. Fc receptors have been found to be crucial for normal immunity and to play an important role in the mechanisms of action of antibody therapeutics. This knowledge is key in designing and optimizing novel therapeutic strategies for human disease. PMID- 19808049 TI - Generation of gut-homing T cells and their localization to the small intestinal mucosa. PMID- 19808050 TI - WITHDRAWN: Communication between immune cells mediated by membrane connections. AB - This article has been withdrawn at the request of the authors and Guest Editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19808051 TI - On the origin and function of immune 'self' recognition. PMID- 19808052 TI - WITHDRAWN: Genetic infectious susceptibility and TLR defects in human. AB - This article has been withdrawn at the request of the authors and the Guest Editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19808053 TI - Sensing and signaling by the immune system NVVI-Dutch Society for immunology course, Lunteren, April 2-3, 2009. PMID- 19808054 TI - Innate recognition of viruses. AB - Innate recognition of viruses is mediated by a panel of intracellular receptors that sense viral genomic nucleic acids and/or replication intermediates. Recognition of these viral signatures takes place in endosomal compartments of specialised cell types via a particular set of Toll-like receptors upon uptake of viral particles or material from infected dying cells. In addition, ubiquitously expressed nucleic acid-sensing molecules in the cytoplasm allow infected cells to detect virus replication and to alert neighbouring cells. This article gives an overview over the different viral nucleic acid sensors and their contribution to the orchestration of anti-viral immune responses. PMID- 19808055 TI - Dusting the sugar fingerprint: C-type lectin signaling in adaptive immunity. AB - Pathogen recognition by dendritic cells (DCs) is central to the induction of adaptive immunity. Pattern recognition receptors (PRRs) on DCs interact with pathogens, leading to signaling events that dictate adaptive immune responses. It is becoming clear that C-type lectins are important PRRs that recognize carbohydrate structures. Most pathogens express carbohydrate structures on their surface and these function as a so-called sugar fingerprint that are recognized by specific C-type lectins. Triggering of C-type lectin induces signaling cascades that initiate or modulate specific cytokine responses and therefore tailor T cell polarization to the pathogens. Here we will discuss our current understanding of the innate signaling pathways induced by C-type lectins DC-SIGN and dectin-1 in humans and how these pathways shape adaptive immunity. PMID- 19808056 TI - Clinical management of HIV-1 resistance. AB - Antiretroviral drug resistance is a fundamental survival strategy for the virus that stems from its vast capacity to generate diversity. With the recent availability of new ARV drugs and classes, it is now possible to prescribe fully active ART to most HIV-infected subjects and achieve viral suppression even in those with multidrug-resistant HIV. It is uncertain, however, if this scenario will endure. Given that ART must be given for life, and new compounds other than second-generation integrase inhibitors may not reach the clinic soon, all efforts must be done to avoid the development of resistance to the new agents. Here, we discuss relevant aspects for the clinical management of antiretroviral drug resistance, leaving detailed explanations of mechanisms and mutation patterns to other articles in this issue. This article forms part of a special issue of Antiviral Research marking the 25th anniversary of antiretroviral drug discovery and development, vol. 85, issue 1, 2010. PMID- 19808057 TI - HIV reservoirs, latency, and reactivation: prospects for eradication. AB - Current antiretroviral therapy effectively suppresses but does not eradicate HIV 1 infection. During therapy patients maintain a persistent low-level viremia requiring lifelong adherence to antiretroviral therapies. This viremia may arise from latently infected reservoirs such as resting memory CD4+ T-cells or sanctuary sites where drug penetration is suboptimal. Understanding the mechanisms of HIV latency will help efforts to eradicate the infection. This review examines the dynamics of persistent viremia, viral reservoirs, the mechanisms behind viral latency, and methods to purge the viral reservoirs. This article forms part of a special issue of Antiviral Research marking the 25th anniversary of antiretroviral drug discovery and development, vol. 85, issue 1, 2010. PMID- 19808058 TI - Improvement of a selective media for the isolation of B. anthracis from soils. AB - To prove linkage between an environmental sample and an anthrax case, there must be isolates obtained from both that can be compared. Although Bacillus anthracis is easily isolated from powder samples, isolating it from soil is difficult because of the high bacterial count in it. Formulations of PLET were prepared, inoculated with B. anthracis, B. cereus and B. thuringiensis and examined for growth. Two hundred eighty-three isolates including 23 B. anthracis were placed onto one formulation while MICs against trimethoprim-sulfamethoxazole were determined. The media supported B. anthracis growth at 30 degrees C and inhibited almost all other bacterial growth, including closely-related species. Sensitivity for B. anthracis and selectivity against other Bacillus and against non-Bacillus were 96.8%, 100% and 97.2% respectively. Isolates that grew had MICs >4 and >76 microg mL(-1) against trimethoprim and sulfamethoxazole, respectively. Soils spiked with 10(2)B. anthracis spores and suspended in PLET broth yielded a 6-7 log(10) increase in B. anthracis. Other growth was inhibited. PLET supplemented with sulfamethoxazole (38 microg mL(-1)), trimethoprim (2 microg mL(-1)), polymyxin B (15,000 U L(-1)), and lysozyme (150,000 U L(-1)) can successfully select for B. anthracis and will facilitate agricultural, environmental and forensic investigations of B. anthracis isolates. PMID- 19808059 TI - Relationships between affective states and decision-making. AB - The study investigated relationships between affective states and decision-making performance using the Iowa Gambling Task. Playing the task involves making decisions between four card decks. Two decks yield high gains and high losses, and if chosen continuously these decks lead to net loss. In contrast, the other two decks yield small gains and small losses, but lead to net profit if they are selected continuously. Correlation and regression analyses revealed a positive relationship between trait-anxiety and decision-making performance and a negative relationship between self-reported emotion regulation and decision-making performance. In addition, anticipatory skin conductance responses were related positively to trait-anxiety and negatively to self-reported emotion regulation. These findings support the somatic marker hypothesis suggesting that emotions facilitate decision-making in complex situations that have uncertain future outcomes. PMID- 19808060 TI - The effect of emotional and attentional load on attentional startle modulation. AB - The interactive effects of emotion and attention on attentional startle modulation were investigated in two experiments. Participants performed a discrimination and counting task with two visual stimuli during which acoustic eyeblink startle-eliciting probes were presented at long lead intervals. In Experiment 1, this task was combined with aversive Pavlovian conditioning. In Group Attend CS+, the attended stimulus was followed by an aversive unconditional stimulus (US) and the ignored stimulus was presented alone whereas the ignored stimulus was paired with the US in Group Attend CS-. In Experiment 2, a non aversive reaction time task US replaced the aversive US. Regardless of the conditioning manipulation and consistent with a modality non-specific account of attentional startle modulation, startle magnitude was larger during attended than ignored stimuli in both experiments. Blink latency shortening was differentially affected by the conditioning manipulations suggesting additive effects of conditioning and discrimination and counting task on blink startle. PMID- 19808061 TI - Anhedonia and effort mobilization in dysphoria: reduced cardiovascular response to reward and punishment. AB - Instigated by evidence for reduced responsiveness to reward in depression, the present two studies addressed the question if such anhedonic behavior would also become evident in reduced mobilization of mental effort in terms of cardiovascular reactivity. Undergraduates completed the Center for Epidemiologic Studies-Depression Scale (CES-D) and worked on mental tasks, expecting either no consequence, a performance-contingent reward, or a performance-contingent punishment. Study 1 revealed that participants with low CES-D scores showed high systolic blood pressure reactivity in the punishment condition, whereas participants with high CES-D scores showed low systolic reactivity. Study 2 corroborated this finding for reward: Nondysphoric participants expecting a reward showed higher reactivity of systolic blood pressure and pre-ejection period than participants in the neutral condition or than dysphoric participants. Together, the studies demonstrate that reward insensitivity in (subclinical) depression is also found in cardiovascular reactivity. Furthermore, dysphoric individuals do not respond to punishment either, suggesting a general insensitivity to hedonic consequences. PMID- 19808062 TI - Gene expression profiles in the common marmoset brain determined using a newly developed common marmoset-specific DNA microarray. AB - To facilitate common marmoset brain research, we produced a DNA microarray with 7557 probe sets derived from the common marmoset brain. Gene expression profiles in the frontal lobe, hippocampus, cerebellum and amygdaloid nucleus were then analyzed and the top 100 probe sets expressed in each structure were compared. The three lists for the frontal lobe, hippocampus and amygdaloid nucleus were very similar but the probe sets for the cerebellum demonstrated specific differences. Some of the genes specifically expressed in cerebellum were analyzed by real-time quantitative PCR to verify the DNA microarray results. Of examined genes, 5 showed extremely strong expression in cerebellum in comparison with the other structures. The results of real-time quantitative PCR were well consistent with the microarray findings, validating our newly developed DNA microarray as a useful tool for brain research with the common marmoset. PMID- 19808063 TI - Enhanced activation of the transient receptor potential channel TRPA1 by ajoene, an allicin derivative. AB - TRPA1 is a calcium-permeable, nonselective cation channel expressed in the dorsal root ganglion and trigeminal ganglia nociceptive neurons. It is activated by the pungent compounds in mustard oil (AITC, allyl isothiocyanate), cinnamon (cinnamaldehyde), garlic (allicin), and is believed to mediate the inflammatory actions of environmental irritants and proalgesic agents. Thiosulfinate (allicin) and isothiocyanate (AITC) compounds contain reactive electrophilic chemical groups that react with cysteine residues within the TRPA1 channel N terminus, leading to channel activation. Ajoene also contains reactive electrophilic chemical groups likely to target TRPA1 channel. Here, we have used voltage-clamp recordings to show that TRPA1-responses are enhanced by ajoene application in a Xenopus oocyte expression system. Though ajoene alone did not activate TRPA1, subsequent application of ajoene enhanced the AITC-, allicin- and depolarization induced responses of TRPA1. Moreover, when increasing concentrations of ajoene were applied along with constant concentrations of allicin or AITC, stronger responses were elicited. These findings suggest that ajoene is a novel TRPA1 channel enhancer, operating in a channel-opening-dependent manner. PMID- 19808064 TI - Role of dual leucine zipper-bearing kinase (DLK/MUK/ZPK) in axonal growth. AB - In developing cerebral cortices, post-mitotic neurons migrate toward the pial surface, elongating their axons concurrently. It has been reported that targeted deletion of the dual leucine zipper-bearing kinase (DLK)/mitogen-activated protein kinase upstream protein kinase (MUK)/leucine-zipper protein kinase (ZPK) gene, which encodes a MAP kinase kinase kinase (MAPKKK) for c-Jun N-terminal kinase (JNK), leads to a neuronal migration-defect and hypoplasia of axonal fiber tracts including those of the anterior commissure and corpus callosum. However, there is no evidence that DLK directly regulates axonal development, because another possibility, i.e. that the defective axonal development in the DLK mutant might be caused secondary to migration failure cannot be ruled out. In this study, we first examined the distributions of DLK mRNA and its protein in the developing cerebral cortex, and found that major portion of DLK proteins appear to be transported into axons. Using dissociated cortical neurons and PC12 cells, we provide direct evidence that DLK regulates axonal elongation. Furthermore, knock-down of DLK decreased the phosphorylation of JNK and its substrate, microtubule-associated protein 1B (MAP1B), which is known to be involved in axonal elongation. These results suggest that the DLK/MUK/ZPK-JNK pathway directly regulates axonal growth through phosphorylation of MAP1B. PMID- 19808065 TI - Adult rat mesenchymal stem cells differentiate into neuronal-like phenotype and express a variety of neuro-regulatory molecules in vitro. AB - Bone marrow stromal stem cells (MSCs), which normally differentiate into mesenchymal derivatives, have recently reported to trans-differentiate into neurons. However, the findings from different groups and interpretations have been challenged. The purpose of this paper is to re-evaluate the phenomenon of neuronal trans-differentiation of MSCs and compare the expression levels of neurotrophins in rMSCs and neuronal-like phenotypes derived from rMSCs. We put rMSCs in 2-mercaptoethanol and 2% dimethylsulfoxide for 5h. Then, the cells were transferred to neuronal induction media composed of DMEM+10%FBS, 10ug/L basic fibroblast growth factor, 10ug/L human epidermal growth factor, 1mmol dibutyryl cyclicn AMP and 0.5mmol isobutylmethylxanthine for 7 days and 14 days. The study demonstrated that the level of BDNF, NGF, NT3, CNTF and GDNF of rMSCs is remarkably higher in rMSCs than the neuronal-like phenotypes, especially CNTF. The expression level of these neurotrophins did not change significantly after enduring induction. We believed that rMSCs can trans-differentiate into neuronal like phenotype under certain conditions. The non-induced rMSCs has a dynamic expression profile of neurotrophins and may serves as a better tool than the trans-differentiated rMSCs for transplant therapy. PMID- 19808066 TI - Draxin, a repulsive axon guidance protein, is involved in hippocampal development. AB - The hippocampus plays an essential role in learning and memory and is one of the major sites implicated in neural diseases. The proper organization of the hippocampus during development is important for its function. We found that draxin, a repulsive axon guidance cue, was widely expressed in the developing hippocampus and draxin deficient mice possessed a smaller hippocampus, particularly in the anterior part of the structure. Quantification of this reduction revealed that the volume of the dentate gyrus of the mutant was significantly smaller compared to the normal counterpart. This size reduction seemed to be dependent on apoptosis rather than due to a decrease in the rate of cell division. PMID- 19808067 TI - Inoculum size-dependent interactive regulation of metabolism and stress response of Saccharomyces cerevisiae revealed by comparative metabolomics. AB - To investigate the metabolic regulation against inoculum density and stress response to high cell density, comparative metabolomic analysis was employed on Saccharomyces cerevisiae under fermentations with five different inoculum sizes by gas chromatography time-of-flight mass spectrometry. Samples from these fermentations were clearly distinguished by principal components analysis, indicating that inoculum size had a profound effect on the metabolism of S. cerevisiae. Potential biomarkers responsible for the discrimination were identified as glycerol, phosphoric acid, succinate, glycine, isoleucine, proline, palmitoleic acid, myo-inositol and ethanolamine. It indicated that enhanced stress protectants in glycerol biosynthesis and amino acid metabolism, depressed citric acid cycle intermediates, as well as decreased metabolites relating to membrane structure and function were involved as the inoculum size of yeast increased. Furthermore, significantly higher levels of glycerol and proline in yeast cells of higher inoculum size fermentation (40 g l(-1)) revealed that they played important roles in protecting yeast from stresses in high cell density fermentation. These findings provided new insights into characterizing the metabolic regulation and stress response depending on inoculum density during ethanol fermentation. PMID- 19808068 TI - Silk gland sericin protein membranes: fabrication and characterization for potential biotechnological applications. AB - This study describes the potential use of silk gland sericin protein as a biocompatible natural biopolymer in its native form. The membranes were fabricated using native silk sericin protein extracted from middle silk gland of Antheraea mylitta, a non-mulberry tropical tasar silkworm without using any cross linking agent. The fabricated membranes were biophysically characterized and optimized for cell culture. Silk sericin protein extracted from gland contained higher amount of beta-sheets, which increased upon treatment with ethanol as observed by FTIR and XRD. The membranes did show robustness, good mechanical strength and high temperature stability. Cytocompatibility of the membranes was evaluated by MTT assay and cell cycle analysis using feline fibroblast cells. Morphology of growing cells was assessed by confocal microscopy that indicated normal spreading and proliferation on the silk sericin membranes. The membranes showed low inflammatory response as observed assaying TNF alpha release. This study reveals the potential of native silk sericin protein from silk gland as biocompatible biopolymer for potential biomedical applications. PMID- 19808069 TI - Breakout session summary from AAPS/CRS joint workshop on critical variables in the in vitro and in vivo performance of parenteral sustained release products. AB - Parenteral drug delivery systems can be designed to provide the flexible delivery characteristics needed in an evolving therapeutic landscape. The goal of some parenteral formulations is to maintain effective drug concentrations over a period of months or years, thereby enhancing patient compliance. When functioning as intended, these formulations can be used to minimize undesirable effects that may occur in response to the fluctuating drug concentrations effected by immediate release products. In other cases, targeted parenteral delivery systems allow for the deposition of drug directly to its site of action, thereby minimizing systemic toxicity. While these novel formulations can be beneficial to both human and veterinary patients, disastrous effects can occur if there is an unanticipated change in product quality or performance. With these thoughts in mind, the Controlled Release Society (CRS) hosted a 2007 workshop entitled "In Vitro and In Vivo Considerations Associated with Parenteral Sustained Release Products". The objective of that workshop was to explore the physicochemical properties of parenteral products and the factors that could alter their in vitro and in vivo performance characteristics. The outcomes of that workshop were summarized in a Journal of Controlled Release article. In response to questions raised during that workshop, the CRS and the American Association of Pharmaceutical Scientist (AAPS) co-hosted the follow-up 2008 workshop entitled "Critical Variables in the In Vitro and In Vivo Performance of Parenteral Sustained Release Products". This 2008 workshop provided a platform for exploring the application of design space concepts to these complex pharmaceuticals, and to consider the corresponding in vitro test methods that can be used to set batch release specifications. To foster discussion, the workshop provided two afternoon breakout sessions where critical questions were explored. This manuscript captures the results of those discussions. PMID- 19808070 TI - A conceptual model of how US families with athletic adolescent daughters manage food and eating. AB - Health professionals concerned about the risks of adolescent obesity and disordered eating practices need greater understanding of how families with adolescents manage food in today's fast paced environment. This paper sought to gain conceptual understanding of the food and eating routines of families with a female adolescent athlete from the perspectives of mothers and daughters. Ten white, non-Hispanic mothers and their daughters were purposively sampled from high school track and cross country teams in Upstate New York. Informants completed in-depth, qualitative interviews. Researchers used the constant comparative method to analyze transcripts for emergent themes and to build a conceptual framework that represented the many factors and processes involved in the construction of family food routines. Families varied in forms and patterns of family eating activities with mothers playing a pivotal role in these routines. Family members' individual needs and values were negotiated in constructing these routines. In this sample the daughters' involvement in sports influenced family eating routines, but mothers' employment, ethnicity, social support, income, and areas of residence also played a role. The model describes how individual participants' food choice processes interact to produce family food routines. The conceptual model can inform research and practice related to the family environments in which adolescents experience food and eating. PMID- 19808071 TI - "Is this a meal or snack?" Situational cues that drive perceptions. AB - What determines whether a person perceives an eating occasion as a meal or snack? The answer may influence what and how much they eat on that occasion and over the remainder of the day. A survey of 122 participants indicated that they used food cues (such as the food quality, portion size, perceived healthfulness, and preparation time) as well as environmental cues (such as the presence of friends and family, whether one is seated, and the quality of napkins and plates) to determine if they were eating a meal rather than a snack. Implications for dieters and for health professionals are provided. PMID- 19808072 TI - CAPA-peptides of praying mantids (Mantodea). AB - Dictyoptera which consist of cockroaches, termites, and praying mantids are among the oldest pterygote insects known. Whereas the localization and sequences of neuropeptides from a number of cockroaches are very well known, nearly nothing is known about the neuropeptides typical of praying mantids. In this study, the neuroanatomy of the median neuroendocrine system in the abdominal ventral nerve cord and the sequences of the CAPA-peptides which are expressed in the respective neuroendocrine cells were analyzed. Altogether, 40 species belonging to different families of Mantodea were included. In contrast to cockroaches, the mantids mostly express two CAPA-periviscerokinins (PVKs), only in Mantis religiosa a third PVK was identified. These PVKs are sequence-related to the PVKs of basal cockroaches (Polyphagidae). In a group of closely related Mantodea (Paramantinae), extended forms of PVK-2 were observed. As shown, these forms are possibly the result of substitutions in the N-terminal cleavage sites of the respective PVKs. No trace of a CAPA-pyrokinin was found in any of the praying mantids. PMID- 19808073 TI - Inhalable powder formulation of a stabilized vasoactive intestinal peptide (VIP) derivative: anti-inflammatory effect in experimental asthmatic rats. AB - Vasoactive intestinal peptide (VIP) exerts immunomodulating and anti-inflammatory activities through its specific receptors, such as VPAC1 and 2 receptors. Previously, a stabilized VIP derivative, [R(15,20,21), L(17)]-VIP-GRR (IK312532), was proposed as a candidate of anti-asthma drug, and a dry powder inhaler system of IK312532 was also developed for inhalation therapy with minimal systemic side effects. In the present study, the anti-inflammatory properties of IK312532 respirable powder (RP) were characterized in an asthma/COPD-like animal model, with the use of newly developed ovalbumin (OVA)-RP for lung inflammation. Marked inflammatory events in the lung were observed after OVA-RP challenge in rats as evidenced by significant increase of inflammatory biomarkers such as eosinophil peroxidase (EPO), myeloperoxidase (MPO) and lactate dehydrogenase (LDH). However, intratracheal administration of IK312532-RP led to significant attenuation of plasma EPO, MPO and LDH activities, as well as significant reduction of recruited inflammatory cells in BALF, especially macrophages and eosinophils. In the rats pretreated with IK312532-RP, histochemical examinations revealed that the inflammatory cells infiltrating to the lung and the epithelial wall thickness decreased significantly by 85% and 58%, respectively. Thus, inhalable powder formulation of IK312532 exerts its anti-inflammatory activity by suppressing granulocyte recruitment to the lung and epithelial hyperplasia, followed by the reduction of cytotoxic peroxidases. PMID- 19808074 TI - Improved Luminex-based human leukocyte antigen-specific antibody screening using dithiothreitol-treated sera. AB - Solid-phase binding assays using purified human leukocyte antigen (HLA) allow the accurate identification and characterization of HLA-specific antibodies in organ transplant patients, but sera may contain factors that block the detection of clinically relevant alloantibodies. The effect of treating patient sera with dithiothreitol (DTT) on alloantibody detection was examined. In 49 sera submitted for routine HLA-specific antibody monitoring, DTT made little difference to immunoglobulin G HLA-specific antibody detection using LABScreen HLA class I and class II mixed beads. However, in sera submitted for antibody identification using single antigen beads (SAB), DTT markedly increased (>twofold increased median fluorescence intensity) antibody binding to HLA class I and/or class II specificities in 14 of 76 (18%) patient sera. In a cohort of highly sensitized patients, treatment of sera with DTT enhanced antibody-binding levels in 14 of 15 (93%) sera. This study highlights the need to consider routine testing of sera with and without DTT for analysis of HLA-specific antibodies by SAB. PMID- 19808075 TI - A regulatory single nucleotide polymorphism in the ubiquitin D gene associated with celiac disease. AB - An aberrant immune response triggered by dietary gluten is the main driving force underlying celiac disease (CD), but other biologic pathways that are dysregulated also participate in disease development. Genetic variation within these pathways might influence expression, contributing to susceptibility to CD. We have investigated the implication of ubiquitin D (UBD), a member of the ubiquitin proteasome system that is strongly upregulated in the intestinal mucosa of active CD. Reverse transcriptase-polymerase chain reaction analysis of intestinal biopsy sample pairs (at diagnosis vs treated) from 30 CD patients confirmed overexpression of UBD in active disease tissue (fold change = 8.3; p = 0.0022). In silico prediction tools identified rs11724 as a putative regulatory single nucleotide polymorphism and association analysis of 468 CD patients and 459 controls revealed that the minor rs11724*C allele was more frequent among patients (minor allele frequency = 0.44 vs 0.39; odds ratio [OR] = 1.23; p = 0.028) and suggested a dominant allele effect (OR = 1.49; p = 0.0045). Correlation of the rs11724 genotype and UBD mRNA levels (OR = 0.76; p = 0.0021) further supports its implication in disease development. PMID- 19808076 TI - Ingestion and excretion of arsenic compounds present in edible brown algae, Hijikia fusiforme, by mice. AB - The element arsenic is a carcinogen and toxic for humans and other living organisms. Some seaweeds contain high amounts of inorganic arsenic (iAs). In particular, Hijikia fusiforme has a high iAs content of approximately 50%. In this study, we examined the absorption, metabolism, excretion, and accumulation of arsenic compounds in mice after the administration of Hijiki. The single-dose experiment, wherein a single dose of cooked Hijiki was administered to the mice, revealed that the urinary and fecal excretion of arsenic compounds was the highest on the first day of dosing, and it became clear that 66-92% of arsenic was excreted within 3 days after administration of the first dose. The repeated dose experiment, wherein repeated doses of cooked or dried Hijiki were administered to the mice, arsenic was detected in all the tissues, but only approximately 5% of the administered dose of arsenic was detected as residual arsenic. These results suggest that the arsenic present in cooked Hijiki is accumulated in very small amounts in mice. PMID- 19808077 TI - The effects of hypervitaminosis A in sheep following intramuscular administrations of vitamin A. AB - Ten ewe lambs (median age 11 months and average weight 29.2+/-2.5 kg) were used in the present study. They were divided into two groups: test (n=5) and control (n=5). Housing and all diets were identical. In the test group vitamin A was injected into the thigh muscle at a daily dose of 5000 IU/kg body weight for 16 days. The average final body weight of sheep in the test group was significantly (P<0.05) less than the control group. All animals were slaughtered at day 17. The lambs' feet were X-rayed to evaluate any difference for radiographic signs between test and control groups. No significant differences were seen for PCV, WBC, differential leukocyte count, and total serum protein between groups. There were no significant differences for serum AST, ALT, and ALP activities and serum calcium, inorganic phosphate, and magnesium concentrations between groups. Histological examination revealed an increased number of lipid droplets in the cytoplasm of the stellate cells of the liver in the test group. The results showed that daily administrations of vitamin A approximately 150 times greater than the daily requirement were well tolerated by sheep. PMID- 19808078 TI - Effect of a single application of TiF(4) and NaF varnishes and solutions on dentin erosion in vitro. AB - OBJECTIVES: This in vitro study aimed to analyse the effect of a single application of TiF(4) and NaF varnishes and solutions to protect against dentin erosion. METHODS: Bovine root dentin samples were pre-treated with NaF-Duraphat varnish (2.26%F, pH 4.5), NaF/CaF(2)-Duofluorid varnish (5.63%F, pH 8.0), NaF experimental varnish (2.45%F, pH 4.5), TiF(4)-experimental varnish (2.45%F, pH 1.2), NaF solution (2.26%F, pH 4.5), TiF(4) solution (2.45%F, pH 1.2) and placebo varnish (pH 5.0, no-F varnish control). Controls remained untreated. Ten samples in each group were then subjected to an erosive demineralisation (Sprite Zero, 4x 90s/day) and remineralisation (artificial saliva, between the erosive cycles) cycling for 5 days. Dentin loss was measured profilometrically after pre treatment and after 1, 3 and 5 days of de-remineralisation cycling. The data were statistically analysed by two-way ANOVA and Bonferroni's post hoc test (p<0.05). RESULTS: After pre-treatment, TiF(4) solution significantly induced surface loss (1.08+/-0.53 microm). Only Duraphat reduced the dentin loss overtime, but it did not significantly differ from placebo varnish (at 3rd and 5th days) and TiF(4) varnish (at 3rd day). CONCLUSIONS: Duraphat varnish seems to be the best option to partially reduce dentin erosion. However, the maintenance of the effects of this treatment after successive erosive challenges is limited. PMID- 19808079 TI - Conformational diseases: looking into the eyes. AB - Conformational diseases, a general term comprising more than 40 disorders are caused by the accumulation of unfolded or misfolded proteins. Improper protein folding (misfolding) as well as accrual of unfolded proteins can lead to the formation of disordered (amorphous) or ordered (amyloid fibril) aggregates. The gradual accumulation of protein aggregates and the acceleration of their formation by stress explain the characteristic late or episodic onset of the diseases. The best studied in this group are neurodegenerative diseases and amyloidosis accompanied by the deposition of a specific aggregation-prone proteins or protein fragments and formation of insoluble fibrils. Amyloidogenic protein accumulation often occurs in the brain tissues, e.g. in Alzheimer's disease with the deposition of amyloid-beta and Tau, in scrapie and bovine spongiform encephalopathy with the accumulation of prion protein, in Parkinson's disease with the deposition of alpha-synuclein. Other examples of amyloid proteins are transthyretin, immunoglobulin light chain, gelsolin, etc. In addition to the brain, the accumulation of unfolded or misfolded proteins leading to pathology takes place in a wide variety of organs and tissues, including different parts of the eye. The best studied ocular conformational diseases are cataract in the lens and retinitis pigmentosa in the retina, but accumulation of misfolded proteins also occurs in other parts of the eye causing various disorders. Furthermore, ocular manifestation of systemic amyloidosis often causes the deposition of amyloidogenic proteins in different ocular tissues. Here we present the data regarding naturally unfolded and misfolded proteins in eye tissues, their structure-function relationships, and molecular mechanisms underlying their involvement in diseases. We also summarize the etiology of ocular conformational diseases and discuss approaches to their treatment. PMID- 19808080 TI - Motion sickness: a negative reinforcement model. AB - Theories pertaining to the "why" of motion sickness are in short supply relative to those detailing the "how." Considering the profoundly disturbing and dysfunctional symptoms of motion sickness, it is difficult to conceive of why this condition is so strongly biologically based in humans and most other mammalian and primate species. It is posited that motion sickness evolved as a potent negative reinforcement system designed to terminate motion involving sensory conflict or postural instability. During our evolution and that of many other species, motion of this type would have impaired evolutionary fitness via injury and/or signaling weakness and vulnerability to predators. The symptoms of motion sickness strongly motivate the individual to terminate the offending motion by early avoidance, cessation of movement, or removal of oneself from the source. The motion sickness negative reinforcement mechanism functions much like pain to strongly motivate evolutionary fitness preserving behavior. Alternative why theories focusing on the elimination of neurotoxins and the discouragement of motion programs yielding vestibular conflict suffer from several problems, foremost that neither can account for the rarity of motion sickness in infants and toddlers. The negative reinforcement model proposed here readily accounts for the absence of motion sickness in infants and toddlers, in that providing strong motivation to terminate aberrant motion does not make sense until a child is old enough to act on this motivation. PMID- 19808081 TI - A new flavonol glycoside from Millettia speciosa. AB - A new flavonol triglycoside, millettiaspecoside D, was isolated from the caulis of Millettia speciosa Champ. Its structure was elucidated on the basis of spectroscopic analysis. PMID- 19808082 TI - Angiotensin II-induced p53-dependent cardiac apoptotic cell death: its prevention by metallothionein. AB - Apoptotic cell death was found to play a critical role in the development of diabetic cardiomyopathy. As one of pathogenic components of diabetes angiotensin II (Ang II) induced cardiac cell death in vitro and in vivo through induction of reactive oxygen and nitrogen species. However, Ang II-induced cell death signaling in the heart remains unclear. The present study was to investigate whether Ang II induces p53 expression and activation and if so, whether Ang II induced cardiac cell death is p53-dependent, and whether a potent antioxidant metallothionein (MT) prevents Ang II-induced p53 expression, and associate apoptotic cell death signaling. A cardiac cell line (H9c2) was exposed to Ang II. We found that exposure of H9c2 cells to Ang II at 10, 50 and 100 nM for 24 h induced a significant apoptotic effect, measured by DNA fragmentation and cleaved caspase-3. Induction of apoptotic cell death by Ang II can be completely blocked by p53 inhibitor Pitithrin-alpha. Exposure of H9c2 cells to Ang II also significantly increased p53 phosphorylation, DNA double strand breaks and Bax/Bcl 2 ratio. All these effects were not observed in H9c2MT7 cells that forcedly overexpresses human MT-IIA gene, suggesting the preventive effect of antioxidant MT against Ang II-induced p53 activation and its apoptotic death signaling. Furthermore, the in vitro finding was validated in animal models by supplying Ang II to wild-type mice (WT) and MT-TG mice that has cardiac-specifically overexpressed MT gene. Ang II-induced significant up-regulation of p53 expression along with an increase in Bax/Bcl-2 ratio in the hearts of WT mice, but not MT-TG mice. These results suggest that Ang II-induced cardiac apoptotic cell death is mediated by p53 apoptotic signaling pathway, which is related to oxidative stress. Antioxidant MT can completely prevent Ang II-induced p53 activation and associated apoptotic effect in the heart. PMID- 19808083 TI - Pharmacological mechanisms underlying the vascular activities of Loranthus ferrugineus Roxb. in rat thoracic aorta. AB - AIM OF THE STUDY: The present study was aimed to investigate the pharmacological basis for the use of Loranthus ferrugineus in hypertension. MATERIALS AND METHODS: Loranthus ferrugineus methanol extract (LFME) was obtained using Soxhelt extractor and then successively fractionated using chloroform, ethyl acetate and n-butanol. The n-butanol fraction of LFME (NBF-LFME) was studied using isolated rat thoracic aorta. RESULTS: NBF-LFME (1.0 x 10(-5) to 3.0mg/ml) was found to be the most potent to concentration-dependently relax the endothelium-intact phenyephrine (PE, 1 microM)- and high K(+) (80 mM)-precontracted rat aortic rings. Removal of the endothelium completely abolished the vascular relaxing properties of NBF-LFME. Pretreatment with atropine (1 microM), L-NAME (10 microM), indomethacin (10 microM) and methylene blue (10 microM) significantly blocked NBF-LFME-mediated relaxation. Endothelium-dependent and -independent relaxations induced by acetylcholine (ACh) and sodium nitroprusside (SNP), respectively, were significantly enhanced in aortic rings pretreated with NBF LFME when compared to those observed in control aortic rings. On the contrary, glibenclamide (10 microM), propranolol (1 microM) and prazosin (0.01 microM) did not alter NBF-LFME-induced relaxation. CONCLUSIONS: The results suggest that NBF LFME induced vascular relaxation by stimulating muscarinic receptors, activating the endothelium-derived nitric oxide-cGMP-relaxant pathway, promoting prostacyclin release and/or possibly through its ability to lengthen the released nitric oxide half-life. The present data further supports previous in vivo findings and explain the traditional use of Loranthus ferrugineus as an anti hypertensive agent. PMID- 19808084 TI - Inhibition of angiotensin convertin enzyme (ACE) activity by the anthocyanins delphinidin- and cyanidin-3-O-sambubiosides from Hibiscus sabdariffa. AB - ETHNOPHARMACOLOGICAL RELEVANCE: The beverages of Hibiscus sabdariffa calyces are widely used in Mexico as diuretic, for treating gastrointestinal disorders, liver diseases, fever, hypercholesterolemia and hypertension. Different works have demonstrated that Hibiscus sabdariffa extracts reduce blood pressure in humans, and recently, we demonstrated that this effect is due to angiotensin converting enzyme (ACE) inhibitor activity. AIM OF THE STUDY: The aim of the current study was to isolate and characterizer the constituents responsible of the ACE activity of the aqueous extract of Hibiscus sabdariffa. MATERIALS AND METHODS: Bioassay guided fractionation of the aqueous extract of dried calyces of Hibiscus sabdariffa using preparative reversed-phase HPLC, and the in vitro ACE Inhibition assay, as biological monitor model, were used for the isolation. The isolated compounds were characterized by spectroscopic methods. RESULTS: The anthocyanins delphinidin-3-O-sambubioside (1) and cyanidin-3-O-sambubioside (2) were isolated by bioassay-guided purification. These compounds showed IC(50) values (84.5 and 68.4 microg/mL, respectively), which are similar to those obtained by related flavonoid glycosides. Kinetic determinations suggested that these compounds inhibit the enzyme activity by competing with the substrate for the active site. CONCLUSIONS: The competitive ACE inhibitor activity of the anthocyanins 1 and 2 is reported for the first time. This activity is in good agreement with the folk medicinal use of Hibiscus sabdariffa calyces as antihypertensive. PMID- 19808085 TI - Transdermal behaviour of the N-alkylamide spilanthol (affinin) from Spilanthes acmella (Compositae) extracts. AB - AIM OF THE STUDY: N-Alkylamides are a large group of bioactive molecules found in several plants from the genera Echinacea, Xanthoxylum and Spilanthes. Extracts and formulations derived from these plants are not only orally used, but also applied on the skin as well. However, there is currently no specific information available about the intrinsic local pharmacokinetics of N-alkylamides after topical application on human skin, questioning the role of this mode of administration. The present study investigates the transdermal behaviour of spilanthol, a prominent N-alkylamide. MATERIALS AND METHODS: Two pharmaceutically accepted dose solutions (ethanol and propylene glycol based aqueous donor vehicles), combined with three different receptor fluids (PBS, PBS+0.5% HPbetaCD, EtOH/H(2)O (30:70, v/v)), were applied on split-thickness human skin in a Franz diffusion cell (FDC) system. Fundamental permeation characteristics of spilanthol in a solvent-independent way (100% aqueous dose solution) were also obtained using an extrapolation approach with different organic solvent/H(2)O ratios. RESULTS AND CONCLUSIONS: We demonstrated for the first time that spilanthol permeates the skin. The following aqueous-extrapolated primary transdermal parameters were obtained (mean+/-SEM): K(p,aq)=3.31 (+/-0.29)x10(-3)cm/h, D(m,aq)=1.86 (+/-0.09)x10(-4)cm(2)/h and K(m,aq)=7.28 (+/-1.59)x10(-1). Partitioning (K(m)) was strongly dependent on the donor solution composition, while diffusion (D(m)) was mainly influenced by the receptor fluid composition. PMID- 19808086 TI - Cognitive enhancement and neuroprotective effects of Bacopa monnieri in Alzheimer's disease model. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Bacopa monnieri (L.) Wettst., a plant belonging to the family Scrophulariaceae, has been used in the traditional system of Ayurvedic medicine to improve intelligence and memory for a long time. Therefore, the potential of this plant to protect against Alzheimer's disease has been raised but less supported document is available. AIM OF THE STUDY: To determine the effect of alcoholic extract of Bacopa monnieri on cognitive function and neurodegeneration in animal model of Alzheimer's disease induced by ethylcholine aziridinium ion (AF64A). MATERIALS AND METHODS: Male Wistar rats were orally given the alcoholic extract of Bacopa monnieri at doses of 20, 40 and 80 mg/kg BW via feeding needle for a period of 2 weeks before and 1 week after the intracerebroventricular administration of AF64A bilaterally. Rats were tested for spatial memory using Morris water maze test and the density of neurons and cholinergic neurons was determined using histological techniques 7 days after AF64A administration. RESULTS: Bacopa monnieri extract improved the escape latency time (p<.01) in Morris water maze test. Moreover, the reduction of neurons and cholinergic neuron densities were also mitigated. CONCLUSION: These findings suggest that Bacopa monnieri is a potential cognitive enhancer and neuroprotectant against Alzheimer's disease. PMID- 19808087 TI - Anti-inflammatory and antinociceptive effects of Baccharis dracunculifolia DC (Asteraceae) in different experimental models. AB - AIM OF THE STUDY: The aerial parts of Baccharis dracunculifolia D.C., popularly known as "alecrim do campo", are used in folk medicine as anti-inflammatory. The aim of the present study was to evaluate the anti-inflammatory and antinociceptive activities of the crude hydroalcoholic extract obtained from leaves of Baccharis dracunculifolia (BdE), which have not been reported. MATERIALS AND METHODS: BdE was analyzed by HPLC and in vivo evaluated (doses ranging from 50 to 400mg/kg, p.o.) by using the acetic acid-induced abdominal constrictions, paw oedema induced by carrageenan or histamine, overt nociception models using capsaicin, glutamate or phorbol myristate acetate (PMA), formalin induced nociception and mechanical hypernociception induced by carrageenan or complete Freund adjuvant (CFA). As positive controls it was used paracetamol in both acetic acid and formalin tests; dipyrone in capsaicin, glutamate and PMA induced nociception; indomethacin in CFA and carrageenan-induced hypernociception models. In addition, the in vitro effects of BdE on COX-2 activity and on the activation of NF-kappaB were also evaluated. RESULTS: BdE (50-400mg/kg, p.o.) significantly diminished the abdominal constrictions induced by acetic acid, glutamate and CFA. Furthermore, BdE also inhibited the nociceptive responses in both phases of formalin-induced nociception. BdE, administered orally, also produced a long-lasting anti-hypernociceptive effect in the acute model of inflammatory pain induced by carrageenan. It was also observed the inhibition of COX-2 activity by BdE. CONCLUSION: In summary, the data reported in this work confirmed the traditional anti-inflammatory indications of Baccharis dracunculifolia leaves and provided biological evidences that Baccharis dracunculifolia, like Brazilian green propolis, possess antinociceptive and anti inflammatory activities. PMID- 19808088 TI - Decline in oxygen consumption correlates with lifespan in long-lived and short lived mutants of Caenorhabditis elegans. AB - In humans, the basal energy metabolism is thought to decline linearly with age. On the other hand, in the nematode Caenorhabditis elegans, two research groups reported independently that it declined exponentially. In this study, furthermore, we used various lifespan-mutant strains to determine whether the previous conclusion is more likely to be true. We can indirectly estimate the metabolic energy by conveniently measuring the oxygen consumption rates of C. elegans using an optical apparatus. From the profile of respiratory rates as a function of age, we can quantitatively isolate the physiological decline rate, lambda, that exponentially represents the decay rate of respiratory activity with age. In addition, quantitative analysis indicates that the respiratory activity of worms has a finite value in advanced age. We also show that the maximum and mean lifespans strongly correlate with the reciprocal of the lambda. These findings offer crucial biochemical evidence for a molecular mechanism at work in biological aging. Consequently, we here propose a mechanism based on a chemical reaction and offer a definition of the physiological decline rate and the finiteness of respiratory activity in advanced age. PMID- 19808089 TI - Effects of social housing of sexually mature male cynomolgus monkeys during general and reproductive toxicity evaluation. AB - We investigated the effects of social housing on reproductive parameters and body weight in mature cynomolgus monkeys, the predominant nonhuman primate model in (reproductive) toxicology. Group housing (n=7) delayed body weight gain compared to single housing. Testicular volumes decreased to approx. 45% of baseline within 13 weeks in low ranking animals followed by return to baseline during weeks 21 26. Interestingly, ejaculate and endocrine parameters did not exhibit corresponding changes. Following separation of this group into a high rank group (n=4) and low rank group (n=3), testicular volumes varied within background variation (+/-15%). Re-allocation of new animals with prior contact/group housing experience into groups of 3 animals also prevented body weight and testicular effects. In conclusion, group formation markedly, albeit transiently, altered body weights and testicular size, and these effects could largely be avoided by previous social interaction for at least 26 weeks. These findings should be considered during social housing of mature animals in toxicity studies. PMID- 19808090 TI - When does the sex ratio of offspring of the paternal 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) exposure decrease: in the spermatozoa stage or at fertilization? AB - Recent animal experiments confirmed that paternal 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) exposure decreases the sex ratio of offspring at birth without altering litter size. However, the timing of this decrease remained unclear. Male mice were administered TCDD at 7-12 weeks of age and mated with non-treated females. The Y-bearing/X-bearing sperm ratio was examined by real-time PCR and FISH methods, and the sex ratio of the 2-cell embryos collected from non-treated females that had been mated with TCDD-exposed males were investigated by nested PCR. The Y-bearing/X-bearing sperm ratio was not significantly decreased in the TCDD group. However, the sex ratio of the 2-cell embryos of the TCDD group was significantly lower than that of the control group. These results may have resulted from a decrease in fertility of Y-bearing sperm. Thus, the results of this study suggested that the sex ratio of the offspring was decreased at fertilization and not during the spermatozoa stage. PMID- 19808091 TI - A modular one-generation reproduction study as a flexible testing system for regulatory safety assessment. AB - The European Union's Registration, Evaluation and Authorisation of Chemicals (REACH) legislation mandates testing and evaluation of approximately 30,000 existing substances within a short period of time, beginning with the most widely used "high production volume" (HPV) chemicals. REACH testing requirements for the roughly 3000 HPV chemicals specify three separate tests for reproductive toxicity: two developmental toxicity studies on different animal species (OECD Test Guideline 414) and a two-generation reproduction toxicity study (OECD TG 416). These studies are highly costly in both economic and animal welfare terms. OECD TG 416 is a fertility study intended to evaluate reproductive performance of animals in the P and F1-generations following repeated exposure to a test substance. It can also be used to detect adverse effects on structural and functional development. Thus, it has conventionally been preferred to the one generation study (OECD TG 415). Recently, the Agricultural Chemical Safety Assessment (ACSA) Technical Committee of the ILSI Health and Environmental Sciences Institute (HESI) proposed that routine two-generation studies could in most cases be replaced with an "enhanced" one-generation study (Reuter et al. [1]). The flexible design proposed by HESI-ACSA allows for the addition of one or more specialised modules, if triggered (e.g. production of a second generation or the investigation of classical developmental toxicity or developmental neuro- or immunotoxicity). Significantly, however, the HESI-ACSA proposal was designed for use in the safety assessment of pesticidal, as opposed to industrial, chemicals. Thus for the purposes of REACH, a streamlined one-generation study that also examines structural development would be the most efficient means of addressing current information requirements for HPV chemicals. This study represents a flexible testing system that can be modified to meet regulatory needs in a variety of sectors. PMID- 19808092 TI - Organization of cholinergic, putative catecholaminergic and serotonergic nuclei in the diencephalon, midbrain and pons of sub-adult male giraffes. AB - The current study describes the nuclear organization and neuronal morphology of the cholinergic, putative catecholaminergic and serotonergic systems within the diencephalon, midbrain and pons of the giraffe using immunohistochemistry for choline acetyltransferase, tyrosine hydroxylase and serotonin. The giraffe has a unique phenotype (the long neck), a large brain (over 500 g) and is a non domesticated animal, while previous studies examining the brains of other Artiodactyls have all been undertaken on domesticated animals. The aim of the present study was to investigate possible differences in the nuclear organization and neuronal morphology of the above-mentioned systems compared to that seen in other Artiodactyls and mammals. The nuclear organization of all three systems within the giraffe brain was similar to that of other Artiodactyls. Some features of interest were noted for the giraffe and in comparison to other mammals studied. The cholinergic neuronal somata of the laterodorsal tegmental nucleus were slightly larger than those of the pedunculopontine tegmental nucleus, a feature not described in other mammals. The putative catecholaminergic system of the giraffe appeared to lack an A15 dorsal nucleus, which is commonly seen in other mammals but absent in the Artiodactyls, had a large and expanded substantia nigra pars reticulata (A9 ventral), a small diffuse portion of the locus coerueleus (A6d), an expansive subcoeruleus (A7sc and A7d), and lacked the A4 nucleus of the locus coeruleus complex. The nuclear organization of the serotonergic system of the giraffe was identical to that seen in all other eutherian mammals studied to date. These observations in the giraffe demonstrate that despite significant changes in life history, phenotype, brain size and time of divergence, species within the same order show the same nuclear organization of the systems investigated. PMID- 19808093 TI - Leberagin-C, A disintegrin-like/cysteine-rich protein from Macrovipera lebetina transmediterranea venom, inhibits alphavbeta3 integrin-mediated cell adhesion. AB - Leberagin-C, a new member of the disintegrin-like/cysteine-rich (D/C) family, was purified to homogeneity from the venom of Tunisian snake Macrovipera lebetina transmediterranea. It is a monomeric protein with a molecular mass of 25,787 Da. Its complete sequence of 205 amino acid residues was established by cDNA cloning. The leberagin-C shows many conserved sequences with other known D/C proteins, like the SECD binding sites and a pattern of 28 cysteines. It is the first purified protein from M. lebetina transmediterranea with only two disintegrin like/cysteine-rich domains. Leberagin-C is able to inhibit platelet aggregation induced by thrombin and arachidonic acid with IC(50) of 40 and 50 nM respectively. It was also able to inhibit the adhesion of melanoma tumour cells on fibrinogen and fibronectin, by interfering with the function of alphavbeta3 and, to a lesser extent, with alphavbeta6 and alpha5beta1 integrins. To our knowledge, leberagin-C is the sole described D/C protein that does not specifically interact with the alpha2beta1 integrin. Structure-activity relationship study of leberagin-C suggested that there are some important amino acid differences with jararhagin, the most studied PIII metalloprotease from Bothrops jararaca, notably around the SECD motif in its disintegrin-like domain. Other regions implicated in leberagin-C specificities could not be excluded. PMID- 19808094 TI - Effects of the special extract ERr 731 from Rheum rhaponticum on estrogen regulated targets in the uterotrophy model of ovariectomized rats. AB - A recent clinical study with a two-year application of the extract ERr 731 from Rheum rhaponticum demonstrated its efficacy and potentially suggested it safety regarding unwanted endometrial side effects. The aim of the present study is to provide experimental proof for the latter observation in a preclinical experimental animal model by assessing dose-dependent effects of ERr 731 - either alone or in combination with estradiol (E2) - on growth and proliferation in the uterus of ovariectomized (ovx) rats. ERr 731 was given in a dose corresponding to human therapeutic application and additionally in three pharmacologically relevant doses. In addition to uterine wet weight, this study examines the effects of ERr 731 on the uterine mRNA expression of the proliferation marker Ki67, proliferating cell nuclear antigen (PCNA), insulin-like growth factor-1 (IGF-1), type 1 IGF receptor (IGF-1R), the two estrogen receptor (ER) subtypes alpha and beta (ERalpha and ERbeta) and the estrogen-responsive gene complement C3 (C3). ERr 731 did neither stimulate an uterotrophic response in the uterotrophic assay with ovx rats nor stimulate or modulate the expression of genes associated with proliferation. In combination with E2, ERr 731 reduced the E2-induced uterine growth stimulation. These observations were further substantiated by the expression pattern of genes related to proliferation control, in view of the fact that the E2-induced elevation of Ki67 mRNA and PCNA protein levels in the uterus were counteracted by simultaneous treatment of the animals with ERr 731. In conclusion, the experimental findings presented here provide further evidence for the safety of ERr 731 towards unwanted uterine and endometrial proliferative events in response to ERr 731 and support observations from recent clinical trials. PMID- 19808096 TI - Evaluation of plasma and tissue estrogen suppression with third-generation aromatase inhibitors: of relevance to clinical understanding? AB - Development of aromatase inhibition and aromatase inhibitors as a therapeutic strategy was initiated through two different pathways. The one pathway went through systematic exploration of aromatase substrate analogues for enzyme inhibitions, subsequently leading to the development of steroidal agents for clinical use. The second involved clinical observation with an unsuccessful anti epileptic compound named aminoglutethimide, attempting to achieve a "medical adrenalectomy". Endocrine studies on patients treated with aminoglutethimide lead to direct assessment of in vivo aromatase inhibition in patients on treatment, thus identifying a novel therapeutic strategy. As such, both research programs represent different examples of pioneering translational work leading towards a successful therapeutic strategy. Subsequent studies with respect to total aromatase inhibition have led to successful development of more potent strategies. Most importantly, these studies have revealed a correlation between aromatase inhibition and clinical outcome. Ongoing studies exploring tissue estrogen levels as well as gene expression profiles on therapy may further improve this important therapeutic area. PMID- 19808095 TI - X-ray structure of human aromatase reveals an androgen-specific active site. AB - Aromatase is a unique cytochrome P450 that catalyzes the removal of the 19-methyl group and aromatization of the A-ring of androgens for the synthesis of estrogens. All human estrogens are synthesized via this enzymatic aromatization pathway. Aromatase inhibitors thus constitute a frontline therapy for estrogen dependent breast cancer. Despite decades of intense investigation, this enzyme of the endoplasmic reticulum membrane has eluded all structure determination efforts. We have determined the crystal structure of the highly active aromatase purified from human placenta, in complex with its natural substrate androstenedione. The structure shows the binding mode of androstenedione in the catalytically active oxidized high-spin ferric state of the enzyme. Hydrogen bond forming interactions and tight packing hydrophobic side chains that complement the puckering of the steroid backbone provide the molecular basis for the exclusive androgenic specificity of aromatase. Locations of catalytic residues and water molecules shed new light on the mechanism of the aromatization step. The structure also suggests a membrane integration model indicative of the passage of steroids through the lipid bilayer. PMID- 19808097 TI - Global diversification of mangrove fauna: a molecular phylogeny of Littoraria (Gastropoda: Littorinidae). AB - The genus Littoraria is one of very few molluscan groups that are closely associated with mangroves. We document its global evolutionary radiation and compare biogeographic patterns with those of mangrove plants, based on phylogenetic and fossil evidence. Using sequences from three genes (nuclear 28S rRNA, mitochondrial 12S rRNA and COI) we reconstruct a phylogeny of 37 of the 39 living morphospecies. Six monophyletic subgenera are defined (Bulimilittorina, Lamellilitorina, Littoraria, Palustorina, Protolittoraria, Littorinopsis) and we synonymize L. coccinea and L. glabrata. A deep division between Palustorina from the Indo-West Pacific and Littoraria from the Atlantic and Eastern Pacific is estimated by a Bayesian relaxed-clock method to be of Middle Eocene to Palaeocene age (43.2-62.7 Ma), which far predates the Early Miocene (18 Ma) closure of the Tethyan Seaway; this, as in mangrove plants, may reflect vicariance by climatic cooling, rather than tectonic processes. The age of Littoraria angulifera in the Atlantic is, however, consistent with Early Miocene vicariance of a Tethyan ancestor. We infer that speciation events are mainly of Miocene or older age, and that diversification has not been driven by depletion of mangrove habitats during recent glacial intervals. Parsimonious reconstruction of ancestral habitats suggests that the genus has inhabited mangrove or wood substrates since its origin, while the rock-dwelling habit of the four members of Protolittoraria is derived. Three species span the Eastern Pacific Barrier, and one is amphi Atlantic, consistent with a long larval phase of up to 10 weeks. Allopatric speciation is inferred, but usually with subsequent range overlap. Ovoviviparity (interpreted as an adaptation to life in mangroves) has arisen twice. PMID- 19808098 TI - Human frontal midline theta and its synchronization to gamma during a verbal delayed match to sample task. AB - The involvement of oscillatory activity, especially at theta and gamma frequency, in human working memory has been reported frequently. A salient pattern during working memory is electroencephalographic frontal midline theta activity which has been suggested to reflect monitoring functions in order to deal with a task. In general, theta activity has been credited with integrative functions of distributed activity. In the present study, we focused on electroencephalographic power analyses and cross-frequency phase synchronization in order to test whether frontal midline theta activity is linked to more locally generated gamma oscillations during the performance of a verbal delayed match to sample task. The task consisted of two different conditions where subjects either had to reorganize three consonant letters in alphabetical order (manipulation condition) or where they merely had to retain the three consonant letters (retention condition). Results revealed higher frontal midline theta activity for the manipulation of maintained stimulus material compared to pure retention of stimulus material. Interestingly, power differences between conditions were most pronounced during the second half of the delay period. Cross-frequency phase synchronization between frontal midline theta activity and distributed gamma activity, on the other hand, was predominant during the first half of the delay period and was stronger for manipulation compared to retention. We suggest that coupling of frontal midline theta to gamma activity reflects monitoring functions on the temporal segregation of memory items, whereas higher frontal midline theta power in the second half of the delay period might be associated with rehearsal processes. Rehearsal processes in the manipulation condition are likely more pronounced, because rehearsal of a new letter string in a limited time window requires higher mental effort compared to pure retention where rehearsal processes may already start at the beginning of the delay period. PMID- 19808099 TI - Molecular characterization of a phospholipid-hydroperoxide glutathione peroxidase from the bumblebee Bombus ignitus. AB - Phospholipid-hydroperoxide glutathione peroxidase (PHGPx or GPx4; EC 1.11.1.12) is an antioxidant enzyme that reduces lipid hydroperoxides in biomembranes. Here, we cloned and characterized cys-PHGPx from the bumblebee Bombus ignitus (Bi PHGPx). The Bi-PHGPx gene consists of 4 exons, encoding 168 amino acid residues with a canonical cys-codon at residue 45 and active site residues Gln(82) and Trp(134). Recombinant Bi-PHGPx, expressed as a 19 kDa protein in baculovirus infected insect cells, exhibited enzymatic activity against PLPC-OOH and H(2)O(2) using glutathione as an electron donor. Tissue distribution analyses showed the presence of Bi-PHGPx in all tissues examined. Bi-PHGPx transcripts were upregulated by stresses, such as wounding, H(2)O(2) exposure, external temperature shock, and starvation. Under H(2)O(2) overload, the RNA interference (RNAi)-induced thioredoxin peroxidase (BiTPx1)-knock-down B. ignitus worker bees showed upregulated expression of Bi-PHGPx in the fat body. These results indicate that Bi-PHGPx is a stress-inducible antioxidant enzyme that acts on phospholipid hydroperoxide and H(2)O(2). PMID- 19808100 TI - Shuttling of SOX proteins. AB - The control of access of SOX proteins to their nuclear target genes is a powerful strategy to activate or repress complex genetic programs. The sub-cellular targeting sequences of SOX proteins are concentrated within the DNA binding motif, the HMG (for high mobility group) domain. Each SOX protein displays two different nuclear localization signals located at the N-terminal and C-terminal part of their highly conserved DNA binding domain. The N-terminal nuclear localization signal binds calmodulin and is potentially regulated by intracellular calcium signalling, while the C-terminal nuclear localization signal, which binds importin-beta, responds to other signalling pathways such as cyclic AMP/protein kinase A. Mutations inducing developmental disorders like sex reversal have been reported in both NLSs of SRY, interfering with its nuclear localization and suggesting that both functional nuclear localization signal are required for its nuclear activity. A nuclear export signal is also present in the HMG box of SOX proteins. Group E SOX proteins harbour a perfect consensus nuclear export signal sequence in contrast to all other SOX proteins, which display only imperfect ones. However, observations made during mouse embryonic development suggest that non-group E SOX proteins could also be regulated by a nuclear export mechanism. The presence of nuclear localization and nuclear export signal sequences confers nucleocytoplasmic shuttling properties to SOX proteins, and suggests that cellular events regulated by SOX proteins are highly dynamic. PMID- 19808101 TI - Thyroid hormone and atherosclerosis. AB - It is generally accepted that the euthyroid state is preferred for the cardiovascular system because both hyperthyroidism and hypothyroidism cause or accelerate cardiovascular diseases. And hypothyroidism is known to be associated with atherosclerosis and ischemic heart diseases. The accelerated atherosclerosis in hypothyroid state has been traditionally ascribed to atherogenic lipid profile, diastolic hypertension and impaired endothelial function. In addition, recent studies suggest that hypothyroidism is associated with the emerging risk factors for atherosclerosis such as hyperhomocysteinemia and an increase in C reactive protein level. Thyroid hormone also has direct anti-atherosclerotic effects such as blood vessel dilatation, production of vasodilatory molecules, and inhibition of angiotensin II receptor expression and its signal transduction. These data suggest that thyroid hormone inhibits atherogenesis through direct effects on the vasculature as well as modifying risk factors for atherosclerosis. This review summarizes the basic and clinical studies on the role of thyroid hormone in atherogenesis and a possible application of thyroid hormone mimetics for the therapy of hypercholesterolemia and atherosclerosis. PMID- 19808102 TI - Mutation in the myelin proteolipid protein gene alters BK and SK channel function in the caudal medulla. AB - Proteolipid protein (Plp) gene mutation in rodents causes severe CNS dysmyelination, early death, and lethal hypoxic ventilatory depression (Miller et al., 2004). To determine if Plp mutation alters neuronal function critical for control of breathing, the nucleus tractus solitarii (nTS) of four rodent strains were studied: myelin deficient rats (MD), myelin synthesis deficient (Plp(msd)), and Plp(null) mice, as well as shiverer (Mbp(shi)) mice, a myelin basic protein mutant. Current-voltage relationships were analyzed using whole-cell patch-clamp in 300 microm brainstem slices. Voltage steps were applied, and inward and outward currents quantified. MD, Plp(msd), and Plp(null), but not Mbp(shi) neurons exhibited reduced outward current in nTS at P21. Apamin blockade of SK calcium-dependent currents and iberiotoxin blockade of BK calcium-dependent currents in the P21 MD rat demonstrated reduced outward current due to dysfunction of these channels. These results provide evidence that Plp mutation specifically alters neuronal excitability through calcium-dependent potassium channels in nTS. PMID- 19808103 TI - Oculo-dento-digital dysplasia: lack of genotype-phenotype correlation for GJA1 mutations and usefulness of neuro-imaging. AB - Oculo-dento-digital dysplasia (ODDD) is an autosomal dominant disorder with complete penetrance and high intra- and interfamilial phenotypic variability. The key features in this syndrome are microphthalmia, enamel hypoplasia and syndactyly of the 4th-5th fingers. ODDD is caused by mutations in the connexin 43 gene (GJA1). We report here four patients from three families with GJA1 mutations, one of them diagnosed prenatally. The three mutations (c.52T > C/p.Ser18Pro, c.689_690delTA/p.Tyr230CysfsX6, c.442C > G/p.Arg148Gly) have been reported once before. Two patients had white matter hypersignal anomalies, associated in one case with mental retardation, but asymptomatic in the other one, an observation that leads us to discuss systematic neuroradiological imaging for ODDD. One case has optic atrophy, another has hypospadias. The patient carrying a truncating mutation of Cx43 did not have palmoplantar keratoderma, in contradiction with the previously suggested genotype-phenotype correlation between truncating mutation and skin involvement. PMID- 19808104 TI - Maize kernel hardness classification by near infrared (NIR) hyperspectral imaging and multivariate data analysis. AB - The use of near infrared (NIR) hyperspectral imaging and hyperspectral image analysis for distinguishing between hard, intermediate and soft maize kernels from inbred lines was evaluated. NIR hyperspectral images of two sets (12 and 24 kernels) of whole maize kernels were acquired using a Spectral Dimensions MatrixNIR camera with a spectral range of 960-1662 nm and a sisuChema SWIR (short wave infrared) hyperspectral pushbroom imaging system with a spectral range of 1000-2498 nm. Exploratory principal component analysis (PCA) was used on absorbance images to remove background, bad pixels and shading. On the cleaned images, PCA could be used effectively to find histological classes including glassy (hard) and floury (soft) endosperm. PCA illustrated a distinct difference between glassy and floury endosperm along principal component (PC) three on the MatrixNIR and PC two on the sisuChema with two distinguishable clusters. Subsequently partial least squares discriminant analysis (PLS-DA) was applied to build a classification model. The PLS-DA model from the MatrixNIR image (12 kernels) resulted in root mean square error of prediction (RMSEP) value of 0.18. This was repeated on the MatrixNIR image of the 24 kernels which resulted in RMSEP of 0.18. The sisuChema image yielded RMSEP value of 0.29. The reproducible results obtained with the different data sets indicate that the method proposed in this paper has a real potential for future classification uses. PMID- 19808105 TI - Improving the detection of significant factors using ANOVA-PCA by selective reduction of residual variability. AB - Selective elimination of residual error can be used when applying Harrington's ANOVA-PCA in order to improve the capabilities of the method. ANOVA-PCA is sometimes unable to discriminate between levels of a factor when sources of high residual variability are present. In some cases this variability is not random, possesses some structure and is large enough to be responsible for the first principal components calculated by the PCA step in the ANOVA-PCA. This fact sometimes makes it impossible for the interesting variance to be in the first two PCA components. By using the proposed selective residuals elimination procedure, one may improve the ability of the method to detect significant factors as well as have an understanding of the different kinds of residual variance present in the data. Two datasets are used to show how the method is used in order to iteratively detect variance associated with the factors even when it is not initially visible. A permutation method is used to confirm that the observed significance of the factors was not accidental. PMID- 19808106 TI - Extraction of multiple pure component 1H and 13C NMR spectra from two mixtures: novel solution obtained by sparse component analysis-based blind decomposition. AB - Sparse component analysis (SCA) is demonstrated for blind extraction of three pure component spectra from only two measured mixed spectra in (13)C and (1)H nuclear magnetic resonance (NMR) spectroscopy. This appears to be the first time to report such results and that is the first novelty of the paper. Presented concept is general and directly applicable to experimental scenarios that possibly would require use of more than two mixtures. However, it is important to emphasize that number of required mixtures is always less than number of components present in these mixtures. The second novelty is formulation of blind NMR spectra decomposition exploiting sparseness of the pure components in the wavelet basis defined by either Morlet or Mexican hat wavelet. This enabled accurate estimation of the concentration matrix and number of pure components by means of data clustering algorithm and pure components spectra by means of linear programming with constraints from both (1)H and (13)C NMR experimental data. The third novelty is capability of proposed method to estimate number of pure components in demanding underdetermined blind source separation (uBSS) scenario. This is in contrast to majority of the BSS algorithms that assume this information to be known in advance. Presented results are important for the NMR spectroscopy-associated data analysis in pharmaceutical industry, medicine diagnostics and natural products research. PMID- 19808107 TI - Determination of sub-micromolar amounts of sulfide by standard free anodic stripping voltammetry and anodic stripping voltammetric titration. AB - A novel electrochemical methods namely standard free anodic stripping voltammetry and anodic stripping voltammetric titration are proposed for determination of dissolved sulfide concentration. 2Ag(+) + S(2-) --> Ag(2)S reaction is used to provide the information. The anodic stripping voltammetric response of unreacted silver-ions at the glassy carbon electrode is used as analytical signal. Results reliability and accuracy are confirmed by analysis of model solutions, spiked natural and tap waters and recovery study, with a recovery of 100 +/- 5% (n=7) obtained. The approaches show the detection limit (3sigma(blank)) of 2-5x10(-10) mol L(-1) and the relative standard deviation of 2-5% for repeated measurements. PMID- 19808108 TI - Comparative studies of praseodymium(III) selective sensors based on newly synthesized Schiff's bases. AB - Praseodymium ion selective polyvinyl chloride (PVC) membrane sensors, based on two new Schiff's bases 1,3-diphenylpropane-1,3-diylidenebis(azan-1 ylidene)diphenol (M(1)) and N,N'-bis(pyridoxylideneiminato) ethylene (M(2)) have been developed and studied. The sensor having membrane composition of PVC: o NPOE: ionophore (M(1)): NaTPB (w/w; mg) of 150: 300: 8: 5 showed best performances in comparison to M(2) based membranes. The sensor based on (M(1)) exhibits the working concentration range 1.0x10(-8) to 1.0x10(-2) M with a detection limit of 5.0x10(-9) M and a Nernstian slope 20.0+/-0.3 mV decade(-1) of activity. It exhibited a quick response time as <8 s and its potential responses were pH independent across the range of 3.5-8.5. The influence of the membrane composition and possible interfering ions have also been investigated on the response properties of the electrode. The sensor has been found to work satisfactorily in partially non-aqueous media up to 15% (v/v) content of methanol, ethanol or acetonitrile and could be used for a period of 3 months. The selectivity coefficients determined by using fixed interference method (FIM) indicate high selectivity for praseodymium(III) ions over wide variety of other cations. To asses its analytical applicability the prepared sensor was successfully applied for determination of praseodymium(III) in spiked water samples. PMID- 19808109 TI - Electrochemical immunoassay using magnetic beads for the determination of zearalenone in baby food: an anticipated analytical tool for food safety. AB - In this work, electrochemical immunoassay involving magnetic beads to determine zearalenone in selected food samples has been developed. The immunoassay scheme has been based on a direct competitive immunoassay method in which antibody coated magnetic beads were employed as the immobilisation support and horseradish peroxidase (HRP) was used as enzymatic label. Amperometric detection has been achieved through the addition of hydrogen peroxide substrate and hydroquinone as mediator. Analytical performance of the electrochemical immunoassay has been evaluated by analysis of maize certified reference material (CRM) and selected baby food samples. A detection limit (LOD) of 0.011 microg L(-1) and EC(50) 0.079 microg L(-1) were obtained allowing the assessment of the detection of zearalenone mycotoxin. In addition, an excellent accuracy with a high recovery yield ranging between 95 and 108% has been obtained. The analytical features have shown the proposed electrochemical immunoassay to be a very powerful and timely screening tool for the food safety scene. PMID- 19808110 TI - Ligandless-dispersive liquid-liquid microextraction of trace amount of copper ions. AB - In the present work, a new ligandless-dispersive liquid-liquid microextraction (LL-DLLME) method has been developed for preconcentration trace amounts of copper as a prior step to its determination by flame atomic absorption spectrometry. In the proposed approach 1,2-dicholorobenzene and ethanol were used as extraction and dispersive solvents, respectively. Some factors influencing on the extraction efficiency of copper and its subsequent determination were studied and optimized, such as the extraction and dispersive solvent type and volume, pH of sample solution, extraction time and salting out effect. Under the optimal conditions, the calibration curve was linear in the range of 1.0 ng mL(-1)-0.6 microg mL(-1) of copper with R(2)=0.9985. Detection limit was 0.5 ng mL(-1) in original solution (3S(b)/m) and the relative standard deviation for seven replicate determination of 0.2 microg mL(-1) copper was +/-1.4%. The proposed method has been applied for determination of copper in standard and water samples with satisfactory results. PMID- 19808111 TI - Extraction and mechanism investigation of trace roxithromycin in real water samples by use of ionic liquid-salt aqueous two-phase system. AB - The ionic liquid, as a green solvent, has several advantages over the organic solvents in traditional liquid-liquid extraction. Aqueous two-phase system (ATPS) consisting of a hydrophilic ionic liquid (1-butyl-3-methylimidazolium tetrafluoraborate, [Bmim]BF(4)) and Na(2)CO(3), which is a novel, simple, non toxic and effective sample pretreatment technique coupled with molecular fluorescence spectrophotometry, was developed for the simultaneous separation, enrichment and rapid analysis of roxithromycin. The extraction yield of roxithromycin in [Bmim]BF(4)-Na(2)CO(3) aqueous two-phase system is influenced by the types of salts, concentrations of Na(2)CO(3) and [Bmim]BF(4), as well as the extracting temperature. Under the optimum conditions, the average extraction efficiency is up to 90.7%. The mechanism of ionic liquid-salt ATPS formation was discussed by hydration theory, and the extraction mechanism of the [Bmim]BF(4) salt ATPS was investigated by FT-IR spectroscopy and UV-vis spectroscopy. The results demonstrate that no chemical (bonding) interactions are observed between ionic liquid and roxithromycin, while the nature properties of the roxithromycin are not altered. This method was practical when applied to the analysis of roxithromycin in real water samples with the detection limit of 0.03 microg mL( 1), relative standard deviation (RSD) of 1.9% (n=13), and linear ranges of 1.00 20.00 microg mL(-1). The proposed extraction technique will be promising in the separation of other small biomolecules. PMID- 19808112 TI - HPLC determination of ibuprofen, diclofenac and salicylic acid using hollow fiber based liquid phase microextraction (HF-LPME). AB - This paper describes an extraction method using a polypropylene membrane supporting dihexyl ether (three-phase hollow fiber-based liquid phase microextraction (HF-LPME)) for the analysis of several pharmaceuticals (salicylic acid (SAC), ibuprofen (IBU) and diclofenac (DIC)) followed by a HPLC determination using a monolithic silica type HPLC column, that allows lower retention times than the usual packed columns with adequate resolution. Detection was realized by means of a coupled in series diode array (DAD) and fluorescence (FLD) detectors. HF-LPME is a relatively new technique employed in analytical chemistry for sample pretreatment which offers more selectivity and sensitivity than any traditional extraction technique. Detection limits by DAD are 12, 53 and 40 ng mL(-1) for salicylic acid, diclofenac and ibuprofen, respectively and by FLD 7 and 2 ng mL(-1) for salicylic acid, and ibuprofen. The method has been successfully applied to their direct determination in human urine and the results obtained demonstrated that could be also applied to the determination of the corresponding metabolites. PMID- 19808113 TI - Comparison of digestion methods for ICP-MS determination of trace elements in fish tissues. AB - A comparison is presented of six methods involving nitric acid in conjunction with other reagents to digest three certified marine biological samples (DOLT-3, DORM-3, IAEA-407) and a fish bone homogenate (prepared from Merluccius australis). An inductively coupled plasma-mass spectrometer with an octopole collision cell was used to determine up to 40 elements (Li, B, Na, Mg, Al, K, Ca, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Rb, Sr, Y, Ag, Cd, Cs, Ba, La, Ce, Pr, Nd, Sm, Eu, Gd, Dy, Ho, Er, Tm, Yb, Lu, Pb, Th, U) in the digests. These results were assessed in terms of their accuracy and precision, and a flow chart was developed to aid the selection of the optimal digestion method. Although none of the methods was found to give accurate determinations for all elements in the different reference materials, a relatively simple method involving nitric acid and hydrogen peroxide heated in a domestic microwave oven gave the most acceptable results. PMID- 19808114 TI - Determination of glucose and ethanol after enzymatic hydrolysis and fermentation of biomass using Raman spectroscopy. AB - Raman spectroscopy has been used for the quantitative determination of the conversion efficiency at each step in the production of ethanol from biomass. The method requires little sample preparation; therefore, it is suitable for screening large numbers of biomass samples and reaction conditions in a complex sample matrix. Dilute acid or ammonia-pretreated corn stover was used as a model biomass for these studies. Ammonia pretreatment was suitable for subsequent measurements with Raman spectroscopy, but dilute acid-pretreated corn stover generated a large background signal that surpassed the Raman signal. The background signal is attributed to lignin, which remains in the plant tissue after dilute acid pretreatment. A commercial enzyme mixture was used for the enzymatic hydrolysis of corn stover, and glucose levels were measured with a dispersive 785 nm Raman spectrometer. The glucose detection limit in hydrolysis liquor by Raman spectroscopy was 8 g L(-1). The mean hydrolysis efficiency for three replicate measurements obtained with Raman spectroscopy (86+/-4%) was compared to the result obtained using an enzymatic reaction with UV-vis spectrophotometry detection (78+/-8%). The results indicate good accuracy, as determined using a Student's t-test, and better precision for the Raman spectroscopy measurement relative to the enzymatic detection assay. The detection of glucose in hydrolysis broth by Raman spectroscopy showed no spectral interference, provided the sample was filtered to remove insoluble cellulose prior to analysis. The hydrolysate was further subjected to fermentation to yield ethanol. The detection limit for ethanol in fermentation broth by Raman spectroscopy was found to be 6 g L(-1). Comparison of the fermentation efficiencies measured by Raman spectroscopy (80+/-10%) and gas chromatography mass spectrometry (87+/-9%) were statistically the same. The work demonstrates the utility of Raman spectroscopy for screening the entire conversion process to generate lignocellulosic ethanol. PMID- 19808115 TI - A sensitive resonance light scattering spectrometry of trace Hg2+ with sulfur ion modified gold nanoparticles. AB - A new resonance light scattering (RLS) spectrometric method for mercury ions (Hg(2+)) in aqueous solutions with sulfur ion (S(2-)) modified gold nanoparticles (Au-NPs-S) has been developed in this contribution. It was found that S(2-) at the surface of Au-NPs resulting from the surface modification can interact with Hg(2+) to form very stable S-Hg-S bonds when Hg(2+) concentration is lower than that of S(2-), resulting in the aggregation of Au-NPs-S and causing enhanced RLS signals. The enhanced RLS intensities (DeltaI(RLS)) characterized at 392 nm were found to be proportional to the concentration of Hg(2+) in the range of 0.025 0.25 micromol L(-1) with a detection limit (3sigma) of 0.013 micromol L(-1). Our results showed that this approach has excellent selectivity for Hg(2+) over other substances in aqueous solutions. PMID- 19808116 TI - Zinc oxide-potassium ferricyanide composite thin film matrix for biosensing applications. AB - Thin film of zinc oxide-potassium ferricyanide (ZnO-KFCN) composite has been deposited on indium tin oxide (ITO) coated corning glass using pulsed laser deposition (PLD). The composite thin film electrode has been exploited for amperometric biosensing in a mediator-free electrolyte. The composite matrix has the advantages of high iso-electric point of ZnO along with enhanced electron communication due to the presence of a redox species in the matrix itself. Glucose oxidase (GOx) has been chosen as the model enzyme for studying the application of the developed matrix to biosensing. The sensing response of the bio-electrode, GOx/ZnO-KFCN/ITO/glass, towards glucose was studied using cylic voltammetry (CV) and photometric assay. The bio-electrode exhibits good linearity from 2.78 mM to 11.11 mM glucose concentration. The low value of Michaelis-Menten constant (1.69 mM) indicates an enhanced affinity of the immobilized enzyme towards its substrate. A quassireversible system is obtained with the composite matrix. The results confirm promising application of the ZnO-KFCN composite matrix for amperometric biosensing applications in a mediator-less electrolyte that could lead to the realization of an integrated lab-on-chip device. PMID- 19808117 TI - Chiral salen Mn(III) complex-based enantioselective potentiometric sensor for L mandelic acid. AB - A new enantioselective potentiometric sensor containing chiral salen Mn(III) as the chiral selector was designed for the assay of L-mandelic acid (L-MA). Optimized membrane electrodes displayed linear dynamic range from 1x10(-5) to 1x10(-1) mol L(-1) with a detection limit of 7.2x10(-6) mol L(-1) and a Nernstian response of -58.1+/-0.5 mV decade(-1) towards L-MA within pH range 7.0-10.2. The potentiometric enantioselectivity coefficient (logK(L,D)(Pot)) of this sensor was -4.0, indicating that the chiral salen Mn(III) complex-based electrode exhibited fairly good discrimination toward L-MA over counter isomer D-MA. The mechanism of chiral recognition for L-MA is discussed by using HF/STO-3G calculation method simulation. PMID- 19808118 TI - Simultaneous evaluation of toxicities using a mammalian cell array chip prepared by photocatalytic lithography. AB - A prototype of a mammalian cell array chip was developed on a flat glass surface. A superhydrophilic (water contact angle=5 degrees)/highly hydrophobic (120 degrees) pattern was prepared on a fluorinated polymer-coated glass surface by means of photocatalytic lithography, and A549 (a human alveolar epithelial cell line), Hep G2 (a human hepatoma cell line) and mouse fibroblast 3T3 cells were inoculated onto the superhydrophilic regions. The cell populations were confined in the superhydrophilic regions for at least 24 h and separated from each other for at least one week. Organ-specific toxicity of aflatoxin B(1) and non-specific toxicity of adriamycin were successfully detected by using the cell array chip. PMID- 19808119 TI - Capillary electrochromatography with contactless conductivity detection for the determination of some inorganic and organic cations using monolithic octadecylsilica columns. AB - A fast separation of alkali and alkaline earth metal cations and ammonium was carried out by capillary electrochromatography on monolithic octadecylsilica columns of 15 cm length and 100 microm inner diameter using water/methanol mixtures containing acetic acid as mobile phase. On-column contactless conductivity detection was used for quantification of these non-UV-absorbing species. The method was also extended successfully to the determination of small amines as well as of amino acids, and the separation selectivity was optimized by varying the composition of the mobile phase. Detection limits of about 1 microM were possible for the inorganic cations as well as for the small amines, while the amino acids could be quantified down to about 10 microM. The separation of 12 amino acids was achieved in the relatively short time of 10 min. PMID- 19808120 TI - Rapid LC-MS detection of cyanobacterial hepatotoxins microcystins and nodularins- comparison of columns. AB - Eight reversed-phase columns intended for rapid HPLC were assessed for the separation of thirteen microcystins and nodularins, cyclic peptidic hepatotoxins. The instrumentation consisted of an Agilent Technologies 1200 Rapid Resolution high performance liquid chromatography system coupled to a mass spectrometer, Bruker Daltonics Ultra Performance High Capacity Ion Trap MS (HCT Ultra) with electrospray ionisation (RRLC-ESI-IT-MS). The columns tested were 2-2.1 mm x 50 mm in diameter and length, and contained small particles (1.8-2.7 microm), or monolithic silica supports for fast performance. The shortest total run time achieved was 3 min 15 s including equilibration and injection. Critical microcystin pairs were still resolved. Several columns showed excellent performance. PMID- 19808121 TI - Kindergarten literacy assessment of English Only and English language learner students: An examination of the predictive validity of three phonemic awareness measures. AB - The study assessed the ability of English phonemic awareness measures to predict kindergarten reading performance and determine factors that contributed to growth trajectories on those measures for English Only (EO) and English language learner (ELL) students. Using initial sound fluency (ISF), phoneme segmentation fluency (PSF), and a combined phoneme segmentation task (CPST), students' beginning of kindergarten scores were used to predict end-of-kindergarten Nonsense Word Fluency (NWF) and reading (WRMT-R/NU). Regression analyses revealed that ISF and CPST early in kindergarten predicted variance in NWF and WRMT-R/NU. PSF did not predict reading performance over ISF or CPST. While gender was a significant factor in the growth curves across the measures, results revealed no significant difference for EO and ELL students. PMID- 19808122 TI - The association between high status positions and aggressive behavior in early adolescence. AB - The present study examined the association between high status social positions in early adolescence and both overt and relational aggression. A sample of 234 sixth graders (132 girls, 102 boys) completed peer nominations assessing aggression, social centrality, and coolness. Data analyses were based on all sixth graders who received peer nominations, for a total sample size of 417 students (224 girls, 193 boys). Hierarchical linear modeling was used to examine both within-peer group differences and between-peer group differences. Results indicated relational aggression was related to higher overall social centrality and being nominated as cool even when controlling for overt aggression. Overt aggression appeared to be related to higher overall social centrality and being nominated as cool, but this relationship was much weaker when relational aggression was controlled. Although girls used relational aggression more often and boys used overt aggression more often, the use of relational aggression by either gender was related to membership in high status peer groups and high status within the peer group. Implications of the findings and suggestions for future research are discussed. PMID- 19808123 TI - Curriculum-based measurement oral reading as an indicator of reading achievement: a meta-analysis of the correlational evidence. AB - This meta-analysis summarized the correlational evidence of the association between the CBM Oral Reading measure (R-CBM) and other standardized measures of reading achievement for students in grades 1-6. Potential moderating variables were also examined (source of criterion test, administration format, grade level, length of time, and type of reading subtest score). Results indicated a significant, strong overall correlation among R-CBM and other standardized tests of reading achievement and differences in correlations as a function of source of test, administration format, and reading subtest type. No differences in the magnitude of correlations were found across grade levels. In addition, there was minimal evidence of publication bias. Results are discussed in terms of existing literature and directions for future research. PMID- 19808124 TI - Anthracycline- and/or taxane-resistant breast cancer: results of a literature review to determine the clinical challenges and current treatment trends. AB - BACKGROUND: As many as 30% of women diagnosed with early breast cancer (BC) will eventually progress to or relapse with locally advanced or meta-static BC. Resistance to the commonly used chemotherapies anthracyclines and taxanes, as well as the approval of new pharmacologic options for treating BC, present important clinical, cost-effectiveness, and societal challenges. OBJECTIVES: The aims of this work were as follows: (1) to review published evidence for potential management strategies, particularly with new therapies, for women with resistant, recurrent, or metastatic BC who have been previously treated with anthracyclines and/or taxanes; and (2) to discuss the wider burden of disease on the patient and society, and potential implications for payers and health care decision makers. METHODS: The PubMed database and relevant congress abstract databases were searched to identify clinical data with relevance to the treatment of recurrent or metastatic BC resistant to anthracyclines and/or taxanes. No date limits were applied, and the search was current as of April 17, 2009. No specific inclusion or exclusion criteria were applied; preference was given to Phase II or III clinical trials published within the past 10 years, although older studies were included if they contained data that guides current clinical practice. RESULTS: Sixteen of the most relevant Phase II or III studies were identified for the 4 agents currently approved for use in this setting, including capecitabine alone (2 studies), capecitabine plus docetaxel (2 studies), ixabepilone alone (5 studies), ixabepilone plus capecitabine (3 studies), gemcitabine plus paclitaxel (1 study), and nanoparticle albumin-bound paclitaxel (3 studies), with overall response rates (complete plus partial responses) ranging from 11.5% to 57%. Other relevant studies are discussed for liposomal doxorubicin, docetaxel, paclitaxel, larotaxel, and vinorelbine, as well as for the addition of biologic agents such as trastuzumab, lapatinib, and bevacizumab to ongoing chemotherapeutic regimens in resistant or metastatic BC. However, only 4 studies discussed the cost of care and cost-effectiveness of current treatment options. CONCLUSIONS: Previous research has reported the efficacy of capecitabine, gemcitabine, nanoparticle albumin-bound paclitaxel, and ixabepilone in the treatment of metastatic BC in patients who have already been treated with anthracyclines and/or taxanes. Such patients previously had few treatment options. Ongoing investigations into novel combination regimens with these agents, including combinations with targeted agents, may further build on this progress. Further research is needed to understand the economic implications of these regimens, including the broader societal effects and the value to patients. PMID- 19808125 TI - Risk for nocturnal hypoglycemia with biphasic insulin aspart 30 compared with biphasic human insulin 30 in adults with type 2 diabetes mellitus: a meta analysis. AB - BACKGROUND: Insulin is recommended as a second-line treatment after diet and metformin fail to reach and/or maintain glycemic targets considered to minimize the risk for long-term diabetic complications. Hypoglycemia and the fear of developing hypoglyce-mia, however, remain substantial barriers to the initiation and optimal use of insulin. OBJECTIVE: The aim of this study was to compare biphasic insulin aspart 30 (BIAsp 30) with biphasic human insulin 30 (BHI 30) with respect to glycemic control and the risk for hypoglycemia using a meta analysis of clinical trials comparing these insulins in patients with type 2 diabetes mellitus (T2DM). METHODS: We included all published and unpublished, randomized, controlled trials in adult patients with T2DM (treatment duration > or = 12 weeks) for which individual patient data were available. All clinical databases and local trial registries of Novo Nordisk A/S (Soeborg, Denmark) were searched to identify clinical trials comparing the 2 products. The predefined primary end point of the study was the overall rate of nocturnal hypoglyce-mia (major, minor, and symptoms-only hypoglycemia occurring from 12:00-6:00 AM). Hypoglycemia was analyzed using a negative binomial distribution model, accounting for exposure time. Glycemic end points were analyzed at 12 to 16 weeks of treatment using ANCOVA, adjusting for baseline. Secondary safety end points were the rates of major hypoglycemia (hypoglycemia requiring third-party assistance), minor hypoglycemia (symptoms confirmed by plasma glucose [PG] <3.1 mmol/L), daytime hypoglycemia (major, minor, and symptoms-only hypoglycemia occurring from 6:01 AM-11:59 PM), overall hypoglycemia (the sum of all major, minor, and symptoms-only episodes), and change in weight from baseline to 12 to 16 weeks of treatment. Secondary efficacy end points were changes in glycosylated hemoglobin (HbA(1c)), fasting PG (FPG), postprandial PG increment (averaged over breakfast, lunch, and dinner), and insulin dose. RESULTS: Nine randomized, parallel or crossover trials were included (N = 1674; male sex, 57%; mean [SD] age, 61.0 [10.6] years; body mass index, 26.7 [4.6] kg/m(2); HbA(1c), 8.1% [1.4%]; duration of diabetes, 10.9 [7.9] years). Rates of overall hypoglycemia were not significantly different (rate ratio [RR] = 1.08; 95% CI, 0.94-1.24; P = NS) between treatments. BIAsp 30 had a 50% lower rate of nocturnal hypoglycemia than BHI 30 (RR = 0.50; 95% CI, 0.38-0.67; P < 0.01), whereas the rate of daytime hypoglycemia was 24% lower for BHI 30 (RR = 1.24; 95% CI, 1.08-1.43; P < 0.01). The likelihood of major hypo-glycemia was significantly lower with BIAsp 30 compared with BHI 30 (odds ratio = 0.45; 95% CI, 0.22-0.93; P < 0.05). BIAsp 30 was associated with reduced PPG increment (averaged over breakfast, lunch and dinner) compared with BHI 30 (treatment difference, -0.31; 95% CI, -0.49 to 0.07; P < 0.01). There was a significantly larger reduction in FPG associated with BHI 30 (treatment difference, 0.63; 95% CI, 0.31-0.95; P < 0.01). However, no significant treatment difference was found for HbA(1c) (treatment difference, 0.04; 95% CI, -0.02 to 0.10; P = NS). CONCLUSION: This meta-analysis found BIAsp 30 to be associated with a significantly lower rate of nocturnal and major hypoglycemia, but a significantly increased risk for daytime hypoglycemia, compared with BHI 30 at a similar level of HbA(1c) in patients with T2DM. PMID- 19808126 TI - Results of a meta-analysis comparing the tolerability of lercanidipine and other dihydropyridine calcium channel blockers. AB - BACKGROUND: Results from clinical studies suggest that the dihydropyridine calcium channel blocker (CCB) lercanidipine may be associated with a lower incidence of peripheral edema than are older dihydro-pyridine CCBs. OBJECTIVE: The objective of the present study was to conduct a meta-analysis of published data from randomized controlled trials (RCTs) to assess the relative risk (RR) of dihydropyridine CCB-specific adverse events with lercanidipine versus the older dihydro-pyridine CCBs (first generation: amlodipine, felodipine, and nifedipine), and versus the other lipophilic dihy-dropyridine CCBs (second generation: lacidipine and manidipine). METHODS: A systematic literature search (all years through August 11, 2008) of MEDLINE, EMBASE, and the Cochrane Library was conducted for English-language reports of single- or double-blind RCTs of > or = 4 weeks' duration that compared the tolerability of lercanidipine with other dihydropyridine CCBs in participants with mild (140-159/90-99 mm Hg) to moderate (160-179/100-109 mm Hg) hypertension. RESULTS: Eight RCTs (6 used first generation drugs, and 4 used second-generation drugs) met the criteria for inclusion. Efficacy outcomes for lowering blood pressure did not differ statistically between lercanid-ipine and either generation of medications. Compared with the first generation, lercanidipine was associated with a reduced risk of peripheral edema (52/742 with lercanidipine vs 88/627 with first generation; RR = 0.44 [95% CI, 0.31-0.62]), but not flushing or headache. The frequency of peripheral edema, flushing, and headache did not differ statistically between lercanidi-pine and the second-generation drugs. Study participants were less likely to withdraw from the RCTs because of peripheral edema (RR = 0.24 [95% CI, 0.12-0.47]) or any adverse event (RR = 0.51 [95% CI, 0.33-0.77]) when treated with lercanidipine rather than a drug from the first generation, but not when treated with lercanidipine rather than second-generation drugs. CONCLUSION: In this meta-analysis, lercanidipine was associated with a lower risk of peripheral edema and a lower risk of treatment withdrawal because of peripheral edema than were the first-generation, but not the second generation, dihydropyridine CCBs. PMID- 19808127 TI - Levocetirizine for the treatment of allergic rhinitis and chronic idiopathic urticaria in adults and children. AB - BACKGROUND: Levocetirizine (LCZ) is a second-generation antihistamine that was approved in January 2008 for the relief of symptoms of seasonal allergic rhinitis (SAR), perennial allergic rhinitis (PAR), and chronic idiopathic urticaria (CIU) in adults and children aged > or = 6 years. OBJECTIVES: This article reviews the available literature on the pharmacokinetics and pharmacodynamics, clinical efficacy and tolerability, and effect on quality of life (QoL) of LCZ. METHODS: A search of the English-language literature was performed using the following databases: MEDLINE (1966-February 2009), International Pharmaceutical Abstracts (19 70-February 2009), Database of Abstracts of Reviews of Effectiveness, Cochrane Database of Systematic Reviews, EMBASE Drugs & Pharmacology (1991 February 2009), Blackwell Synergy, CINAHL Plus with Full Text, EBSCOhost, ScienceDirect, and Wiley Interscience. The search terms were levocetirizine, allergic rhinitis, chronic idiopathic urticaria, antihistamine, pharmacokinetics, quality of life, drug interactions, case reports, and cost. Publications describing studies of > or = 2 weeks' duration that concerned the efficacy, tolerability, pharmacoeconomics, and/or QoL effects of LCZ were included in the review. RESULTS: In 4 studies in adult patients with moderate to severe PAR, LCZ 5 mg/d was associated with significant improvements in symptom scores for sneezing, rhinorrhea, and ocular/nasal pruritus at 4 to 6 weeks compared with placebo (P < or = 0.05). In 3 studies, nasal congestion scores were significantly improved within 4 to 6 weeks compared with placebo (P < 0.001). LCZ 5 mg/d was associated with improvements compared with placebo in scores for the ability to do housework, complete work activities, and engage in outdoor activities at 6 months (P < or = 0.011). In a 6-week study in children with moderate to severe SAR, LCZ 5 mg/d was associated with significant improvements compared with placebo in sneezing, rhin-orrhea, and itchy nose (P < 0.004); significant improvements in symptoms from baseline were also seen in a 4-week study in adults with SAR (P < 0.001). One study in patients with SAR reported no significant difference between LCZ and fluticasone compared with fluticasone monotherapy in terms of improvement in QoL, nasal airflow obstruction, sneezing, or pruritus. In a 6-week study in patients with moderate to severe CIU, LCZ 5 mg/d was significantly more effective than placebo in reducing overall CIU symptoms (P < 0.05). In two 4-week studies, one comparing LCZ 5 mg/d with placebo and the other comparing it with desloratadine (DSL), LCZ was significantly more effective than either comparator in terms of improvement in scores for pruritus severity (P < or = 0.001 vs placebo; P < 0.004 vs DSL) and duration (P < or = 0.001 vs placebo; P = 0.009 vs DSL). LCZ was significantly more effective than placebo (but not DSL) in reducing the number and size of wheals (both, P = 0.001). In a 12-week, open label, crossover study, patients reported significantly longer symptom relief with cetirizine than LCZ (P < 0.005). The most commonly reported adverse events in two 6-month studies in adults with PAR treated with LCZ 5 mg/d included headache (23.8%), pharyngitis (19.4%), influenza (14.6%), fatigue (8.3%), and somnolence (8.3%). There is serious concern about the possibility of febrile seizures in infants treated with LCZ. Three pharmacoeconomic studies of LCZ 5 mg/d were identified, one comparing it with placebo in patients with PAR, one comparing it with placebo in patients with CIU, and another comparing it with second-generation antihistamines and montelukast in patients with PAR. Because of design limitations and differences in comparators in these studies, it was not possible to determine the cost-effectiveness of LCZ in the treatment of PAR or CIU. CONCLUSIONS: In the studies reviewed, LCZ 5 mg/d was effective in reducing symptoms of PAR, SAR, and CIU and improving QoL, with an acceptable tolerabili-ty profile. There is a need for studies of longer durations, head-to-head comparisons against other anti-histamines, drug-interaction studies, safety studies in infants, and cost-effectiveness analyses. PMID- 19808128 TI - Neutralization of alpha(2)-antiplasmin by microplasmin: a randomized, double blind, placebo-controlled, ascending-dose study in healthy male volunteers. AB - BACKGROUND: Microplasmin is the isolated proteinase domain of plasmin. Administration of microplasmin has been found to neutralize alpha(2)-antiplasmin (alpha(2)-AP) activity, which has been associated with reduced infarct size in various preclinical models of stroke. OBJECTIVES: The goals of this first-in-man study were to investigate the ability of microplasmin to neutralize alpha(2)-AP activity and to monitor its tolerability in healthy male volunteers. METHODS: This Phase I, double-blind, placebo-controlled, ascending-dose study included 10 groups, each containing 6 subjects who were randomized to receive microplasmin or placebo in a 2:1 ratio. The study had 3 parts. In part 1, subjects received a single intravenous bolus of microplasmin 0.1, 0.5, 1, 1.5, or 2 mg/kg or placebo over 15 minutes. In part 2, subjects received a bolus of microplasmin 1 mg/kg or placebo, followed by an infusion of 1, 2, 3, or 4 mg/kg or placebo over 60 minutes. In part 3, subjects, all of whom were aged >55 years, received a bolus of mi-croplasmin 1 mg/kg or placebo, followed by an infusion of 1 mg/kg or placebo. The primary pharmaco-dynamic end point was the change in alpha(2)-AP activity, measured at different time points up to 4 days after dosing using a chromogenic assay. All adverse events were monitored based on spontaneous reports and nondirected questioning at study visits up to the post-study visit 21 days after administration of study drug. RESULTS: The mean (SD) age of subjects in parts 1, 2, and 3 was 30 (8), 30 (8), and 64 (8) years, respectively. All groups receiving microplasmin had a dose-dependent decrease in alpha(2)-AP activity. In part 1, the mean maximal inhibition of alpha(2)-AP was 11.8% (6.0%), 27.7% (4.3%), 53.0% (4.8%), 65.3% (4.3%), and 84.0% (6.0%) after bolus administration of microplasmin 0.1, 0.5, 1, 1.5, and 2 mg/kg, respectively, and 7.4% (6.9%) after administration of placebo. In part 2, the mean maximal inhibition of alpha(2)-AP was 74.6% (11.2%), 95.5% (3.3%), 99.0% (1.0%), and 88.0% (12.5%) after bolus administration of microplasmin 1 mg/kg followed by an infusion of 1, 2, 3, and 4 mg/kg, respectively, compared with 12.9% (6.8%) after administration of placebo. In part 3, the mean maximal inhibition was 69.7% (3.4%) after bolus administration of microplasmin 1 mg/kg followed by an infusion of 1 mg/kg, compared with 8.8% (4.1%) with placebo. One subject in the highest dose group in part 1 (2 mg/kg) and 2 subjects in the highest dose group in part 2 (1 + 4 mg/kg) had an urticarial reaction. All 3 subjects also had a decrease in total hemolytic complement and an increase in complement 5a. CONCLUSIONS: Neutralization of alpha(2)-AP activity by microplasmin was feasible in these healthy male volunteers. The urticarial reactions observed in the highest dose groups were considered dose-limiting adverse events. Further trials are needed to investigate the tolerability of this therapy and whether it is neuroprotective in patients with acute ischemic stroke. PMID- 19808129 TI - Utilization characteristics and treatment persistence in patients prescribed low dose buprenorphine patches in primary care in the United Kingdom: a retrospective cohort study. AB - BACKGROUND: The 7-day, low-dose buprenorphine patch has been available in the United Kingdom since 2005 for the treatment of chronic nonmalignant pain that is unresponsive to nonopioid analgesics. Osteo-arthritis pain, a significant cause of pain and disability in the elderly, is a common reason for prescribing bu prenorphine patches. OBJECTIVES: The goals of this study were to investigate utilization and treatment persistence in patients receiving low-dose buprenorphine patches and the expected patterns of treatment 12 months after the initiation of treatment. METHODS: This was a retrospective cohort study of patients who were prescribed low-dose buprenorphine patches in general practice in the United Kingdom. Patients in this cohort were matched by age, sex, and practice with comparator cohorts prescribed oral codeine, dihydrocodeine, or tramadol. Data on baseline characteristics, utilization, and adverse events were obtained from the General Practice Research Database, which contains computerized medical records from UK general practice. Treatment persistence was determined based on repeat prescribing within 90 days after the expected end of a prescription; rates of persistence were compared between the buprenorphine and comparator cohorts. Cox proportional hazards regression models were used to compare the incidence of typical opioid adverse effects (constipation, dizziness, and nausea and/or vomiting) between cohorts. RESULTS: The study cohort included 4968 patients who were prescribed low-dose buprenorphine patches. The majority of patients (64.2%) were aged >65 years, and the most frequently recorded indication for low-dose buprenorphine patches was osteoarthritis (48.7%). Most patients (76.1%) started treatment at the lowest patch strength (5 microg/h). The mean patch strength prescribed over time stabilized at 10 to 12 microg/h. Persistence with low-dose buprenorphine patches over 6 months was significantly higher than with codeine, dihydrocodeine, and tramadol (28.9%, 22.4%, 24.4%, and 23.8%, respectively; P < 0.01). Persistence over 12 months also was significantly higher with low-dose buprenorphine patches compared with the comparators (18.5%, 16.1%, 18.0%, and 17.6%; P < 0.01). After 12 months, the difference in persistence levels between cohorts was not statistically significant. In the Cox proportional hazards regression models, patients using buprenorphine patches had an increased incidence of constipation, dizziness, and nausea and vomiting compared with those who used the comparator opioids (P < 0.05). CONCLUSIONS: Significantly more patients receiving low-dose buprenorphine patches in this study persisted with treatment at 6 and 12 months compared with those receiving other opioid analgesics. Treatment with low-dose buprenorphine patches was most frequently initiated at the lowest patch strength and stabilized at a mean of 10 to 12 microg/h. PMID- 19808130 TI - Comparison of the efficacy and safety of arformoterol 15 microg twice daily and arformoterol 30 microg once daily in COPD: a single-dose, multicenter, randomized, modified-blind, two-way crossover study. AB - OBJECTIVE: The aim of this study was to compare the efficacy and safety of nebulized arformoterol 15 microg/2 mL twice daily (ARF15 BID) and 30 microg/4 mL once daily (ARF30 QD) in subjects with moderate to severe chronic obstructive pulmonary disease (COPD). METHODS: In this single-dose, multicenter, randomized, modified-blind, 2-way crossover study, subjects aged > or =45 years with moderate to severe COPD, a forced expiratory volume in 1 second (FEV(1)) > or =0.7 L, and < or =65% predicted FEV1, and a FEV(1):forced vital capacity ratio < or =70% were randomly assigned to receive single-day treatment with ARF15 BID or ARF30 QD in random order, separated by a 5 +/- 2-day washout period. The primary efficacy end point was time-normalized AUC of FEV(1) from baseline (hour 0) to 24 hours (FEV(1)AUC(0-24)). Secondary efficacy end points were time-normalized AUC of FEV(1) from baseline to 12 hours (FEV(1)AUC(0-12)) and from 12 to 24 hours (FEV(1)AUC(12-24)), and FEV(1) at 24 hours after administration of the morning dose (trough FEV(1)). Equivalence of the 2 therapies was assessed by comparing the 90% CI value for the difference of the least squares mean (LSM) to a study specific predefined equivalence range for change in FEV(1)AUC(0-24) of -0.07 to 0.07 L. RESULTS: A total of 33 subjects were enrolled (20 men, 13 women; mean [SD] age, 64.5 [8.8] years; 15 subjects received ARF15 BID first; 18 received ARF30 QD first). ARF15 BID and ARF30 QD were associated with similar improvements from baseline in (FEV(1)AUC(0-24), LSM 0.15 and 0.16 L, respectively; Delta, 0.01 L; 90% CI, -0.02 to 0.04) and trough FEV(1) (LSM, 0.15 and 0.12 L, respectively; Delta, -0.03 L; 90% CI, -0.09 to 0.03). FEV(1)AUC(0-12) was improved more with ARF30 QD than ARF15 BID (Delta, 0.06 L; 90% CI, 0.04 to 0.09), and FEV(1)AUC(12 24) was improved more with ARF15 BID than ARF30 Qd (Delta, -0.04 L; 90% CI, -0.08 to 0.01). The 90% CI for FEV(1)AUC(0-24) for the treatment difference between ARF15 BID and ARF30 QD was within the prespecified range of -0.07 to 0.07 L, indicating that both treatments resulted in equivalent FEV(1)AUC(0-24) values. CONCLUSIONS: In these subjects with moderate to severe COPD, single-day administrations of ARF15 BID or ARF30 QD were associated with FEV(1) responses over a period of 24 hours that were considered equivalent per the protocol definition employed in the present study. FEV1 improvement over 12 hours was greater for ARF30 QD after the morning dose and for ARF15 BID after the evening dose. ClinicalTrials.gov Identifier: NCT00571428. PMID- 19808132 TI - An evidence-based assessment of the clinical significance of drug-drug interactions between disease-modifying antirheumatic drugs and non-antirheumatic drugs according to rheumatologists and pharmacists. AB - BACKGROUND: Clinically relevant drug-drug interactions (DDIs) must be recognized in a timely manner and managed appropriately to prevent adverse drug reactions or therapeutic failure. Because the evidence for most DDIs is based on case reports or poorly documented clinical information, there is a need for better assessment of their clinical relevance. OBJECTIVE: This study evaluates the interdisciplinary agreement between rheumatologists and clinical (hospital) pharmacists in assessing the clinical relevance of DDIs with disease-modifying antirheumatic drugs (DMARDs) and non-DMARD medications. METHODS: Potential DDIs were identified from the medical literature using MEDLINE and EMBASE for the years 1968-2009. The following search terms were used for the key word, title, and abstract sections of the publications: interaction(s), DMARD, disease modifying antirheumatic drug(s), antirheumatic, rheumatology, rheumatoid arthritis, and the names of the individual DMARDs of interest (abatacept, adalimumab, anakinra, auranofin, aurothioglucose, aurothiomalate, d penicillamine, etanercept, gold, [hydroxy]-chloroquine, interleukin-1 receptor antagonist, IL1-RA, infliximab, leflunomide, methotrexate, rituximab, and sulfasalazine/sulphasalazine). Reference lists of the retrieved publications were searched for further information on potential DDIs. All pharmacodynamic or pharmacokinetic DDIs between a DMARD and a non-DMARD identified were included in the study, with the exception of evidence regarding DMARD doses higher than used in the treatment of rheumatoid arthritis and interactions with phytotherapeutic or homeopathic preparations. Using a standard information set for each DDI (eg, from product labeling, textbooks, and the medical literature), a group of rheumatologists and a group of clinical pharmacists independently assessed whether the individual drug-DMARD combinations interacted and whether they required immediate intervention. Both groups consisted of 3 members (2 men and 1 woman), aged 40 to 60 years, who had >5 years of clinical experience and were currently involved in clinical practice in large, nonacademic teaching hospitals in the Netherlands. RESULTS: Forty potential DDIs with DMARDs were retrieved and assessed by the 2 groups. For 30 (75%) of these, rheumatologists and clinical pharmacists agreed about the requirement for immediate intervention. Specifically, 17 drug combinations (43%) were judged to interact and to require immediate intervention, and 13 combinations (33%) were judged either not to interact or to interact but not to require immediate intervention. For 10 combinations (25%), rheumatologists and clinical pharmacists were not in agreement. Overall, agreement between the groups was good (kappa = 0.80) for judging whether the drug combinations were interactions, and agreement was fair (kappa = 0.39) for judging whether immediate intervention was required. Prospective analysis of the data showed that rheumatologists tended to recommend immediate intervention more often when the adverse reaction to the DDI involved an increased risk of toxicity of the DMARD. In contrast, clinical pharmacists more often advocated immediate intervention when the adverse reaction involved decreased effectiveness of the DMARD. CONCLUSION: For a subset of DMARD-drug combinations, rheumatologists and clinical pharmacists differed in their assessments of clinical relevance. PMID- 19808131 TI - Long-term follow-up of the original interferon-beta1b trial in multiple sclerosis: design and lessons from a 16-year observational study. AB - OBJECTIVE: This article describes the design of and difficulties inherent in the execution of a long-term, observational trial that sought to assess the validity of short-term measures of multiple sclerosis (MS) (eg, relapse rate, inflammatory lesions) for long-term disease outcomes. METHODS: In the original double-blind, placebo-controlled interferon (IFN)-p1b study, 372 patients with relapsing remitting MS (Expanded Disability Status Scale score 0.0-5.5) were randomly assigned to IFN-beta1b 50 ug (n = 125), IFN-beta1b 250 microg (n = 124), or placebo (n = 123) for 2 years. These patients were recruited 16 years later for participation in this long-term follow-up (LTF) study, which had no exclusion criteria or drug interventions. RESULTS: The 11 centers identified 88.2% (328/372) of the original study patients at LTF; however, 10.8% (n = 40) refused to participate and 9.4% (n = 35) were deceased. Detailed evaluations were available for 260 patients, which included 7 deceased patients. No differences in demographic or baseline disease characteristics were found between individuals who did and did not participate in the LTF. More patients randomly assigned to placebo in the original trial were deceased (20/123 [16.3%]) than those assigned to IFN-beta1b 50 microg (9/125 [7.2%]; uncorrected P = 0.044) or IFN-beta1b 250 microg (6/124 [4.8%]; uncorrected P = 0.003). CONCLUSIONS: Although most patients (88.2%) were identified at LTF, ascertainment was incomplete. This was attributable to patients' refusal to participate, loss to follow-up, or death. Delays in the registration of death data and recent privacy legislation provided further barriers. Mortality was lower for patients originally randomized to receive IFN-beta1b rather than placebo. We recommend that all short-term trials on chronic diseases include provisions for LTF. PMID- 19808133 TI - Upper gastrointestinal tolerability of alendronate sodium monohydrate 10 mg once daily in postmenopausal women: a 12-week, randomized, double-blind, placebo controlled, exploratory study. AB - BACKGROUND: The dissolution profiles of generic oral bisphosphonate alendronate (ALN) sodium for the treatment of postmenopausal osteoporosis differ by formulation, suggesting potential differences in the risk for upper gastrointestinal (GI) irritation. OBJECTIVE: This study compared the tolerability profile of ALN monohydrate with that of placebo, with a focus on upper GI irritation, in postmenopausal women with osteoporosis. METHODS: This multicenter, double-blind, placebo-controlled estimation study enrolled postmenopausal women with osteoporosis. Patients were randomized in a 2:1 ratio to receive ALN monohydrate 10 mg or placebo once daily for 12 weeks. Tolerability was monitored throughout the study and up to 14 days after administration of the final dose. Primary end points were the proportions of patients with upper GI adverse events (AEs); upper GI AEs that were rated as serious or study drug related or that led to study discontinuation; and esophageal AEs. Between-treatment differences and associated 95% CIs were assessed using the Wilson score method. RESULTS: Of 438 patients who were randomized, 367 (mean age, 65.5 years; history of osteoporotic fracture, 6.8%; ALN monohydrate, 237; placebo, 130) completed the study. The proportion of patients with a history of upper GI disorders at baseline was numerically greater in the ALN monohydrate group than in the placebo group (117 [40.2%] and 45 [30.6%], respectively). The proportions of patients with active baseline upper GI disease were 83 (28.5%) and 30 (20.4%) in the ALN monohydrate and placebo groups, respectively. The proportions of patients who experienced an upper GI AE during the study period were 66 (22.7%) and 30 (20.4%) (95% CI, -6.2 to 10.0). The proportions of patients with upper GI AEs that were rated as serious or study drug related or that led to study discontinuation were 20.3% and 12.9% (95% CI, -0.3% to 14.1%). Three serious AEs in the active-treatment group (breast cancer, 2; wrist fracture, 1) were not considered related to the study drug, nor was the 1 serious AE in the placebo group (wrist fracture). One patient (ALN monohydrate) had an esophageal AE (nonserious spasm). Approximately 8% of patients who received ALN monohydrate reported dyspepsia, compared with none who received placebo. Within each treatment group, the rates of upper GI AEs were numerically higher in patients with a history of upper GI disease. CONCLUSIONS: In these postmenopausal women who received ALN monohydrate or placebo, upper GI AEs were common (20.4%-22.7%). The proportion of patients who experienced upper GI AEs considered drug related or that led to discontinuation was appar- ently greater with ALN monohydrate compared with placebo. PMID- 19808134 TI - Efficacy of roxithromycin in adult patients with rheumatoid arthritis who had not received disease-modifying antirheumatic drugs: a 3-month, randomized, double blind, placebo-controlled trial. AB - BACKGROUND: It has been reported that antibodies to oral anaerobic bacteria are elevated in the serum and synovial fluids of patients with rheumatoid arthritis. Macrolide antibiotics are active against oral anaerobic bacteria. OBJECTIVE: The aim of this work was to evaluate the clinical efficacy of roxithromycin in patients with early seropositive rheumatoid arthritis. METHODS: This was a double blind trial. We enrolled adult patients with early rheumatoid arthritis who had not previously received disease-modifying antirheumatic drugs and randomized them to receive either once-daily oral roxithromycin 300 mg or once-daily oral placebo for 3 months. The primary efficacy variable was the percentage of patients who had a 20% improvement according to the American College of Rheumatology (ACR) criteria (an ACR 20 response) at 3 months. Secondary outcome measures were 50% improvement and 70% improvement according to ACR criteria (an ACR 50 response and an ACR 70 response, respectively). The 28-joint disease activity score (DAS28) was also calculated. Clinical remission was defined as DAS28 score <2.6, and a low level of disease activity was defined as DAS28 score <3.2 but > or =2.6. Adverse event data (eg, example, type, severity, time of occurrence, time to resolution) were obtained from physical examinations and patient self-reporting. RESULTS: The roxithromycin group had 16 patients (mean [SD] age, 45 [4] years; 11 women, 5 men; all white). The placebo group had 15 patients (mean [SD] age, 42 [5] years; 10 women, 5 men; all white). A significantly greater percentage of patients treated with 300 mg of roxithromycin experienced an ACR 20 re- sponse at 3 months, compared with those who received placebo (75% [n = 12] vs 20% [n = 3]; P = 0.002). Greater percentages of patients treated with 300 mg of roxithromycin also achieved ACR 50 responses (56% [n = 9] vs 7% [n = 1]; P = 0.003) and ACR 70 responses (44% [n = 7] vs 0%; P = 0.004) compared with patients who received placebo. At month 3, DAS28 response rates were significantly greater with once daily roxithromycin 300 mg than with once-daily placebo (P < 0.001). Adverse events were reported for 11 patients (69%) in the roxithromycin group and 7 patients (47%) in the placebo group. The most common adverse events (>5%) were nausea, abdominal pain, headache, and dry mouth. There were no dose-limiting toxic effects. One participant in the roxithromycin group withdrew from the study because of severe emesis; two withdrew from the placebo group because of lack of efficacy. CONCLUSIONS: In these adult patients with rheumatoid arthritis, 3-month treatment with roxithromycin significantly improved the signs and symptoms of rheumatoid arthritis and was generally well tolerated. Future studies should investigate the relationship between disease activity and serum or joint antibodies to anaerobic bacteria. PMID- 19808135 TI - Modafinil for the treatment of hypersomnia associated with myotonic muscular dystrophy in adults: a multicenter, prospective, randomized, double-blind, placebo-controlled, 4-week trial. AB - BACKGROUND: Myotonic muscular dystrophy type 1 (MMD1) is the most common form of adult MD, with a mean prevalence of 1 in 8000. Excessive daytime sleepiness (ie, hypersomnia) is a common complication of MMD1. OBJECTIVE: The aim of this study was to evaluate the efficacy and tolerability of modafinil for the treatment of hypersomnia in adults with MMD1. METHODS: This multicenter, prospective, randomized, double-blind, placebo-controlled study consisted of a prerandomization period (90 to 2 days before randomization) and a 4-week randomization period in which patients were assigned to receive either active treatment (modafinil 300 mg/d) or placebo. The study was conducted at 3 clinics in France between February 2000 and June 2002. Adult patients aged > or =18 years, with genetically proven MMD1, an Epworth Sleepiness Scale (ESS) score >10, and a mean latency to sleep onset < or =8 minutes measured by the Multiple Sleep Latency Test (MSLT) were eligible. The primary efficacy end point was the Maintenance of Wakefulness Test (MWT) score at 4 weeks. Secondary end points included the mean MSLT score and scores from the ESS, physician's assessment of the therapeutic effect and the patient's global self-assessment via visual analog scale, the 17-item Hamilton Depression Rating Scale, and the Short Form Health Survey (SF-36) quality-of-life assessment. RESULTS: A total of 28 patients (15 men, 13 women; mean [SD] age, 40 [12.7] years [range, 18-69 years]; 100% white; modafinil group, 13; placebo group, 15) completed the study without protocol violations. Of the 28 patients with MMD1 included in the analysis, 21 had adult onset MMD1. At 4 weeks, the mean MWT score was 16.4 (3.3) minutes in the modafinil group and 15.8 (3.8) minutes in the placebo group (P = NS). At the end of the randomization period, there were no significant between-group differences in any secondary outcome. A total of 8 patients (4 in each group) reported > or =1 adverse event, including digestive, neurologic, and skin symptoms. Weight loss was reported in 1 patient (2 kg). CONCLUSION: In this small study conducted in an adult population with MMD1 and a high prevalence of hyper-somnia, modafinil had no significant effects on daytime somnolence measured using objective MWTs. PMID- 19808136 TI - Pharmacokinetics and tolerability of single escalating doses of gabapentin enacarbil: a randomized-sequence, double-blind, placebo-controlled crossover study in healthy volunteers. AB - BACKGROUND: Gabapentin enacarbil is an actively transported prodrug of gabapentin that provides predictable dose-proportional gabapentin exposure with high (> or =68%) oral bioavailability. OBJECTIVES: The aims of this study were to investigate the pharmacokinetics and tolerability of gabapentin enacarbil up to supratherapeutic doses and the effects of gabapentin enacarbil on cardiac repolarization in healthy volunteers, and to provide a dose reference for a future definitive QT/corrected QT (QTc) study. METHODS: This was a randomized sequence, double-blind, placebo-controlled, single escalating-dose, crossover study of gabapentin enacarbil 600-mg extended-release tablets administered as a single oral dose of 2400, 3600, 4800, or 6000 mg or placebo, with a 1-week washout between administrations. Blood samples were collected over a period of 36 hours after administration and were analyzed using a validated method of liquid chromatography/tandem mass spec-trometry. Blood gabapentin enacarbil and gabapentin concentrations were analyzed using noncompartmental methods. Tolerability was assessed by monitoring adverse events (AEs) (using subject interview/reporting), laboratory parameters, vital sign measurements, and 12-lead electrocardiography (ECG). Holter ECG was also performed. RESULTS: Thirty-two healthy volunteers were included in the study (18 women, 14 men; mean [SD] age, 31.2 [11.4] years; body mass index, 24.9 [3.04] kg/m(2)). Gabapentin enacarbil was converted rapidly to gaba-pentin after absorption. Gabapentin exposure in blood was proportional to gabapentin enacarbil dose over the range of 2400 to 6000 mg (1250-3125 mg-equivalent gabapentin). Blood concentrations of intact gabapen-tin enacarbil were low and transient (< or =0.5% of the released gabapentin concentration at all doses). The most commonly reported AEs were dizziness and nausea (50% and 25% of subjects, respectively). All but 4 AEs were mild to moderate in intensity. Two subjects experienced treatment-emergent AEs rated as severe: psychomotor retardation, vertigo, and sedation (4800-mg dose) and somnolence (6000 mg). All treatment-emergent AEs resolved without medical intervention. No serious AEs were reported, and none of the AEs led to study withdrawal. There were no clinically significant changes in laboratory parameters, vital sign measurements, or ECG values; QTc intervals did not exceed 480 msec or change from baseline >30 msec at any gabapentin enacarbil dose. CONCLUSIONS: Gabapentin enacarbil was associated with dose-proportional gabapentin exposure at doses up to 6000 mg and was generally well tolerated in these healthy subjects. These findings support the use of 6000-mg gabapentin enacarbil in a definitive QT/QTc study. PMID- 19808137 TI - Pharmacokinetic and bioequivalence comparison between orally disintegrating and conventional tablet formulations of flurbiprofen: a single-dose, randomized sequence, open-label, two-period crossover study in healthy Chinese male volunteers. AB - BACKGROUND: Flurbiprofen, an NSAID, is used for the treatment of inflammation and pain caused by rheumatoid arthritis and osteoarthritis as well as soft-tissue injuries. A new orally disintegrating tablet (ODT) of flurbiprofen has recently been developed; this study was conducted to provide support for this drug to obtain marketing authorization in China. OBJECTIVE: The aim of the study was to compare the pharmacokinetic properties and bioequivalence of flurbiprofen 50-mg ODT (test) with a conventional flurbiprofen 50-mg tablet (reference) under fasting conditions in healthy volunteers. METHODS: This was a single-dose, randomized-sequence, open-label, 2-period crossover study. Healthy, nonsmoking Chinese male volunteers were randomly assigned to receive 150 mg (administered as three 50-mg tablets) of either the test or reference formulation of flurbiprofen, followed by a 7-day washout period and administration of the alternate formulation. Study drugs were administered after a 12-hour overnight fast. Blood samples were collected before dosing and at 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8, 12, and 24 hours after dosing. Serum flurbiprofen concentrations were analyzed using a validated nonstereospecific liquid chromatography/tandem mass spectrometry method. Pharmacokinetic parameters, including C(max), T(max), t(1/2), AUC(0-24), and AUC(0-infinity), were calculated and analyzed statistically. C(max), AUC(0-24), and AUC(0-infinity) were used to test for bioequivalence after natural logarithm (ln)-transformation. Tolerability was evaluated throughout the study by clinical assessments, vital sign monitoring, physical examinations, 12-lead ECG, clinical laboratory tests, and questioning subjects about adverse events (AEs). RESULTS: A total of 20 Chinese males (mean [SD] age, 21.4 [2.5] years [range, 19-28 years]; height, 174.4 [4.2] cm [range, 169-183 cm]; weight, 63.2 [5.1] kg [range, 56-78 kg]; body mass index, 20.8 [1.4] kg/m(2) [range, 19-24 kg/m(2)]) completed the study. No period or sequence effect was observed. The 90% CIs for the ln-transformed ratios of Cmax, AUC(0-24), and AUC(0-infinity) were 99.9% to 115.9%, 97.8% to 107.9%, and 100.3% to 110.9%, respectively, meeting the predetermined criteria for bioequivalence. Two subjects (10.0%) experienced 1 of 2 mild AEs (increase in total bilirubin and dizziness), which were not considered to be associated with study drug administration. CONCLUSIONS: This single-dose 150-mg (three 50-mg tablets) study of each formulation of flurbiprofen found that the test and reference products met the regulatory criteria for bioequivalence in these fasting healthy Chinese male volunteers. Both formulations were generally well tolerated. State Food and Drug Administration of China study registration number: 2005L04356. PMID- 19808138 TI - Bioavailability of two oral formulations of a single dose of levofloxacin 500 mg: an open-label, randomized, two-period crossover comparison in healthy Mexican volunteers. AB - BACKGROUND: Levofloxacin is a synthetic fluoroquinolone with a broad spectrum of antibacterial activity. It is indicated for the treatment of respiratory, sinus, skin, and urinary tract infections. Although generic formulations of oral levofloxacin are marketed in Mexico, a literature search did not identify published data concerning the bioavailability of these formulations; these data would be relevant to secure marketing of a test formulation in Mexico. OBJECTIVE: The aim of this study was to compare the bioavailability and determine the bioequivalence of a test formulation (an oral tablet containing levofloxacin 500 mg) with its corresponding listed reference-drug formulation in Mexico (a list issued by Mexican Health Authorities). METHODS: A single-dose, open-label, randomized-sequence, 2-period crossover design was used in this study. Eligible participants were healthy Mexican adults of either sex, randomly assigned to receive the test formulation followed by the corresponding reference formulation, or vice versa, with a 1-week washout period between doses. After a 10-hour overnight fast, the participants received the assigned formulation. Plasma concentrations of levofloxacin were determined using high-performance thin-layer chromatography, and densitometric analysis was performed at 300 nm. For the analysis of pharmacokinetic parameters, including C(max), AUC0(-24), and AUC(0 infinity)), blood samples were drawn at baseline and at 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.5, 3, 4, 6, 8, 12, and 24 hours after administration. The test formulation was considered to meet the criteria for bioequivalence if the geometric mean ratios (test/reference) were with- in the predetermined range of 80% to 125%. Tolerability was determined by clinical assessment, vital signs, laboratory analysis, and interviews with participants about adverse events. RESULTS: A total of 26 participants were enrolled, including 14 men and 12 women with a mean (SD) age of 24 (4) years (range, 18-34 years), weight of 62.2 (10.0) kg (range, 45.5-80.0 kg), height of 163 (9) cm (range, 148-176 cm), and body mass index of 23.3 (2.4) kg/m(2) (range, 19.2-27.1 kg/m(2)). The 90% CIs for log transformed C(max), AUC(0-24), and AUC(0-infinity) were 94.48% to 106.22%, 90.01% to 116.44%, and 85.11% to 114.00%, respectively. Eleven participants reported a total of 20 adverse events during the study. None of the adverse events were considered serious. CONCLUSIONS: In this small study in healthy, fasting Mexican adults, a single 500-mg dose of the test formulation of orally administered levofloxacin met the regulatory requirements to assume bioequivalence based on the rate and extent of absorption. Both formulations were well tolerated. PMID- 19808139 TI - Assessment of the bioequivalence of brand and biogeneric formulations of abciximab for the treatment of acute coronary syndrome: a prospective, open label, randomized, controlled study in Korean patients. AB - BACKGROUND: Abciximab has been found to reduce major adverse cardiovascular events in patients with acute coronary syndrome (ACS). A previous study reported on the tolerability of biogeneric abciximab in patients with ACS. This formulation has been approved by the Korea Food and Drug Administration and is currently being marketed. Its ex vivo antiplatelet effect, however, has not been compared with that of branded abciximab. OBJECTIVE: The purpose of the present study was to compare ex vivo antiplatelet activity, angiographic outcome, and bleeding complications between biogeneric and branded abciximab. METHODS: This prospective, open-label, randomized, controlled study was conducted in Korea. Patients with ACS who underwent percutaneous coronary intervention (PCI) were randomized to receive biogeneric abciximab or branded abciximab. All patients received intracoronary unfractionated heparin 70 IU/kg and either biogeneric or branded abciximab 0.25 mg/kg IV bolus approximately 10 minutes before undergoing PCI, followed by a 0.125 microg/kg/min 12-hour infusion of the same formulation. The antiplatelet effect of both drugs was assessed at 3 time points (at baseline, and 10 minutes and 24 hours after the end of the bolus infusion) using a validated rapid platelet-function assay. RESULTS: In total, 37 patients (30 men and 7 women; 19 receiving biogeneric abciximab and 18 receiving branded abciximab) were included. Patient demographics did not differ significantly between the 2 groups (16 men [84.2%] and 3 women [15.8%] in the biogeneric group vs 14 men [77.8%] and 4 women [22.2%] in the branded group; mean [SD] age, 65 [11] vs 60 [10] years; weight, 64.6 [8.7] vs 67.9 [10.1] kg, respectively). The bolus and the continuous infusion of the biogeneric and branded formulations achieved similar levels of platelet inhibition, with a mean (SD) inhibition of platelet aggregation >90% at 10 minutes after the end of the bolus infusion (94.7% [8.2%] vs 92.6% [16.9%], respectively; P = NS) and >65% at 24 hours (68.1% [9.8%] vs 70.9% [9.7%]; P = NS) compared with baseline. One thrombolysis in myocardial infarction major bleeding complication (retro-peritoneal hemorrhage) was reported in a patient who received biogeneric abciximab. CONCLUSION: There were no statistically significant differences in the antiplatelet effects of these 2 formulations in this small, selected population of Korean patients with ACS. PMID- 19808140 TI - Comparison of temsirolimus pharmacokinetics in patients with renal cell carcinoma not receiving dialysis and those receiving hemodialysis: a case series. AB - BACKGROUND: Intravenous temsirolimus, an inhibitor of the mammalian target of rapamycin (mTOR), is approved for the treatment of advanced renal cell carcinoma (RCC). Sirolimus, the principal metabolite of temsirolimus in humans, also exhibits mTOR inhibitory activity. OBJECTIVE: The purpose of this study was to compare the pharmacokinetics of temsirolimus and its metabolite, sirolimus, among patients with RCC not receiving dialysis and those receiving hemodialysis. METHODS: This was a single-center, unblinded, single-dose study. Patients with histologically confirmed metastatic RCC were eligible. A single 25-mg dose of temsirolimus was administered as a 30-minute intravenous infusion during the first round of chemotherapy. Blood samples were drawn at 0 (predose), 0.5 (end of infusion), 1.5, 2.5, 5.5, 24, 72, and 144 hours after infusion. In patients receiving hemodialysis, an additional blood sample was drawn 1 hour after each treatment to compare pre- and postconcentration. Temsirolimus concentrations were assayed in blood using HPLC coupled to mass spectrometry. Pharmacokinetic parameters (C(max), T(max), t((1/2)), AUC(0-infinity), total body clearance, volume of distribution at steady state, AUC ratio [the ratio of sirolimus to temsirolimus AUCs], and AUC sum [the algebraic sum of temsirolimus and sirolimus AUCs]) were calculated and analyzed statistically. RESULTS: In total, 13 consecutive patients (11 men and 2 women; 11 not receiving dialysis and 2 receiving hemodialysis) were included. No patient refused to participate in the study. Of those not receiving dialysis, the median age was 54 years (range, 36-77 years), and of those receiving hemodialysis, the median age was 60.5 years (60-61 years). There were no significant between-group differences in the pharmacokinetic parameters of temsirolimus and sirolimus. Moreover, in patients receiving hemodialysis, blood drug concentrations assessed immediately before hemodialysis were similar to those assayed 1 hour after the treatment. CONCLUSION: This study found that after single-dose administration of 25 mg of temsirolimus as a 30-minute intravenous infusion, neither temsirolimus nor sirolimus concentrations were significantly affected in these patients with RCC receiving hemodi-alysis compared with those not receiving dialysis. PMID- 19808141 TI - Research on special populations: The need for and promise of pediatric, adolescent, and maternal therapeutics. PMID- 19808142 TI - Management of fever in children: summary of the Italian Pediatric Society guidelines. AB - OBJECTIVE: This article summarizes the Italian Pediatric Society guideline on the management of the signs and symptoms of fever in children, prepared as part of the National Guideline Program (NGLP). METHODS: Relevant publications in English and Italian were identified through searches of MEDLINE and the Cochrane Database of Systematic Reviews from their inception through December 31, 2007. Based on the consensus of a multidisciplinary expert panel, the strength of the recommendations was categorized into 5 grades (A-E) according to NGLP methodology. SUMMARY: In the health care setting, axillary measurement of body temperature using a digital thermometer is recommended in children aged <4 weeks; for children aged > or =4 weeks, axillary measurement using a digital thermometer or tympanic measurement using an infrared thermometer is recommended. When body temperature is measured at home by parents or care-givers, axillary measurement using a digital thermometer is recommended for all children. Children who are afebrile when seen by the clinician but are reported to have had fever by their caregivers should be considered febrile. In special circumstances, high fever may be a predictive factor for severe bacterial infection. Use of physical methods of reducing fever is discouraged, except in the case of hyperthermia. Use of antipyretics-paracetamol (acetaminophen) or ibuprofen-is recommended only when fever is associated with discomfort. Combined or alternating use of antipyretics is discouraged. The dose of antipyretic should be based on the child's weight rather than age. Whenever possible, oral administration of paracetamol is preferable to rectal administration. Use of ibuprofen is not recommended in febrile children with chickenpox or dehydration. Use of ibuprofen or paracetamol is not contraindicated in febrile children with asthma. There is insufficient evidence to form any recommendations concerning fever in children with other chronic conditions, but caution is advised in cases of severe hepatic/renal failure or severe malnutrition. Newborns with fever should always be hospitalized because of the elevated risk of severe disease; paracetamol may be used, with the dose adjusted to gestational age. Use of paracetamol or ibuprofen is not effective in preventing febrile convulsion or the adverse effects of vaccines. PMID- 19808143 TI - Long-term tolerability of the methylphenidate transdermal system in pediatric attention-deficit/hyperactivity disorder: a multicenter, prospective, 12-month, open-label, uncontrolled, phase III extension of four clinical trials. AB - BACKGROUND: Short-term treatment with the meth-ylphenidate transdermal system (MTS) has been well tolerated in several clinical trials in children with attention-deficit/hyperactivity disorder (ADHD). However, the effects of long term use have not been systematically evaluated. OBJECTIVES: The primary objective of this study was to assess the 12-month tolerability of MTS in children with ADHD. Effectiveness was a secondary objective. METHODS: This Phase III study was a multicenter, 12-month, open-label, flexible-dose extension of 4 previous trials. In those studies, children aged 6 to 12 years with a diagnosis of ADHD (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision criteria) received MTS, osmotic-release oral system methylphenidate, or placebo. At entry into the present study, the children either continued to receive their optimal dose of MTS (10, 15, 20, or 30 mg per 9-hour patch wear time) or underwent dose titration over 4 weeks to an optimal MTS dose, which was continued for the remainder of the study. Tolerability was evaluated based on adverse events (AEs), physical examinations, vital signs, electrocardiograms, laboratory tests, the Children's Sleep Habits Questionnaire, and the occurrence of application-site reactions. RESULTS: Of 327 enrolled subjects, 326 received treatment and 157 completed the study. The majority of enrolled subjects were male (64.8%) and white (73.7%), with a mean (SD) age of 9.2 (1.9) years. Two hundred sixty-five (81.3%) of the 326 subjects who received MTS reported AEs. AEs led to study discontinuation in 29 subjects (8.9%). The majority (98.3%) of treatment-emergent AEs were of mild or moderate severity. The most common AEs were decreased appetite (24.8%), headache (16.6%), upper respiratory tract infection (12.3%), cough (11.7%), pyrexia (10.1%), and decreased weight (10.1%). Of the 1118 AEs, 40.8% were considered possibly or probably related to study treatment. Three serious AEs (facial contusion, ankle fracture, and syncope) occurred and were considered unrelated to study treatment. Based on data collected across all study visits, application-site reactions generally consisted of mild erythema associated with mild discomfort at the patch site. Application-site reactions accounted for 22 (6.7%) study discontinuations. CONCLUSIONS: Slightly less than half (48.0%) of subjects completed this 12-month, open-label extension study of MTS. Most AEs were mild to moderate in severity and, with the exception of application-site reactions, were typical of those previously observed with methylphenidate. ClinicalTrials.gov identifier: NCT00151957. PMID- 19808145 TI - Prevalence of the prescribing of potentially inappropriate medications at ambulatory care visits by elderly patients covered by the Taiwanese National Health Insurance program. AB - BACKGROUND: The use of potentially inappropriate medications can have profound medical consequences for elderly patients and place a substantial burden on the health care system. OBJECTIVES: This study was conducted to determine the prevalence of potentially inappropriate medication prescribing at ambulatory care visits by patients aged > or =65 years covered by the Taiwanese National Health Insurance program, to examine the characteristics of and risk factors for such prescribing, and to investigate its influence on health care resource utilization. METHODS: Ambulatory care visits by patients aged > or =65 years in 2001-2004 were identified from the National Health Insurance claims database. The 2003 Beers criteria for drugs to be avoided in the elderly were used to identify potentially inappropriate medications prescribed at these visits. Only drugs with the potential to lead to higher-severity adverse events were included. Multivariate logistic regression was used to determine predictors of the prescribing of potentially inappropriate medications at ambulatory care visits. Independent variables in the regression model included patient characteristics (eg, sex, age), physician characteristics (sex, age, and specialty), and visit characteristics (site and prescribed drug number). The dependent variable was visits that included a prescription for a potentially inappropriate medication. RESULTS: Overall, 176,661,994 ambulatory care visits by patients aged > or =65 years were identified in 2001-2004. Of these, 19.1% involved a prescription for a potentially inappropriate medication. Although the frequency of potentially inappropriate medication prescribing declined over the study period, 62.5% of elderly patients were exposed to such medications in 2004. The only patient characteristic associated with an increased likelihood of the prescribing of potentially inappropriate medications was female sex (male sex: odds ratio [OR] = 0.982 [95% CI, 0.980-0.983], P < 0.001). Physician characteristics associated with a greater likelihood of the prescribing of potentially inappropriate medications was male sex (OR = 1.206 [95% CI, 1.202-1.210], P < 0.001); older age (43-50 years: OR = 1.021 [95% CI, 1.018-1.025], P < 0.001; >/=51 years: OR = 1.238 [95% CI, 1.235-1.242], P < 0.001); and family medicine/general practice (OR = 1.267 [95% CI, 1.265-1.269], P < 0.001). For visit characteristics, significant associations were found with visits to a primary care clinic (OR = 1.887 [95% CI, 1.881-1.892], P < 0.001) and the number of drugs prescribed (4-6 drugs: OR = 2.701 [95% CI, 2.696-2.706], P < 0.001; > or =7 drugs: OR = 4.528 [95% CI, 4.517 4.538], P < 0.001). The most commonly prescribed types of potentially inappropriate medications were antihistamines (4.8% of all prescriptions in 48.3% of elderly patients), muscle relaxants/antispasmodics (4.0% and 40.3%, respectively), and long-acting benzodiazepines (2.4% and 21.4%). In 2004, the mean number of ambulatory care visits per patient was significantly higher among those who received potentially inappropriate medications compared with those who did not (30.78 vs 16.57, respectively; P < 0.001). Patients who received potentially inappropriate medications also had significantly more emergency department visits (0.27 vs 0.15; P < 0.001) and hospital admissions (0.46 vs 0.27; P < 0.001). CONCLUSION: There was a high prevalence of the prescribing of potentially inappropriate medications at ambulatory care visits by elderly patients in Taiwan in 2001-2004. PMID- 19808146 TI - Pattern and predictors of the initiation of biologic agents for the treatment of rheumatoid arthritis in the United States: an analysis using a large observational data bank. AB - OBJECTIVE: The aim of this study was to identify factors associated with the initiation of biologic agents for the treatment of rheumatoid arthritis (RA) in a large US observational cohort. METHODS: Semiannual patient-reported data in the ARAMIS (Arthritis, Rheumatism and Aging Medical Information System) data bank from January 1998 to January 2006 were analyzed retrospectively using pooled logistic regression (with adjustment for center-level and temporal effects) to identify patient-, disease-, and treatment-related characteristics associated with the initiation of biologics for the treatment of RA. RESULTS: The analysis included 1545 patients from 7 US centers. By 2006, 41.4% of 679 patients remaining in the sample had received biologics. Initiation of biologics was significantly associated with greater disability in the previous 6-month period (per 1-unit increase in Health Assessment Questionnaire score: odds ratio [OR] = 1.45; 95% CI, 1.22-1.72; P < 0.01) and treatment in the previous period with steroids (OR = 2.24; 95% CI, 1.76-2.85; P < 0.01) or nonbiologic disease modifying antirheumatic drugs (OR = 2.43; 95% CI, 1.71-3.46; P < 0.01). Two sociodemographic factors were significant predictors of decreased use of biologics: older age (per 10 years: OR = 0.74; 95% CI, 0.660.82; P < 0.01) and lower annual income (per $10,000 reduction: OR = 0.95; 95% CI, 0.91-1.00; P = 0.04). There were no significant differences with respect to sex, race, employment status, comorbidity, previous NSAID use, or treatment center. CONCLUSIONS: Disease- and treatment-related factors were significant predictors of the initiation of biologics for RA. Independent of these factors, however, biologics were less often used in patients who were older and those with lower incomes. Use of biologics increased steadily over the period studied. PMID- 19808147 TI - The recent paper by Daley-Yates et al. PMID- 19808149 TI - Having a CMD is associated with improved nursing home quality of care. PMID- 19808150 TI - Treatment of bacteriuria without urinary signs, symptoms, or systemic infectious illness (S/S/S). PMID- 19808151 TI - The basis for improving and reforming long-term care, part 2: clinical problem solving and evidence-based care. AB - There are intense efforts to improve the quality of long-term care. However, it is unclear whether these efforts are based on understanding root causes of the deficits in quality. This article focuses on processes of clinical problem solving and decision making as a means to enable safe, effective, efficient, and person-centered care that reflects key principles discussed in the initial article in this series. The care delivery process is the means for applying these principles to deliver care. The techniques used in clinical decision-making and problem-solving activities are not unique to health care. Whether or not it is recognized, clinical problem-solving and decision-making activities are occurring continually in all long-term care facilities. But only some staff and practitioners do them well. There is much talk about applying "evidence-based care" in all settings, including the nursing home. However, the term is widely misunderstood and only sometimes applied properly. True evidence-based care requires combining scientific evidence with sufficiently detailed evidence about the individual patient. This article applies the discussion to identify criteria for "expertise" in long-term care. We may identify characteristics of "experts" in long-term care, regardless of discipline, as well as factors that distinguish levels of expertise. Experts have the skill and judgment to apply knowledge effectively to individual patient situations. Based on these criteria, only some of the claims to expertise in caring for, advising about, or overseeing long-term care residents and patients are warranted. PMID- 19808152 TI - Clinical update on nursing home medicine: 2009. AB - This represents the third clinical update based on presentations at the American Medical Directors Association (AMDA) annual meeting. Topics covered this year include hypertension, heart failure, cachexia, dehydration, falls, depression, constipation, and aspiration pneumonia. PMID- 19808153 TI - Gender differences and health status in old and very old patients. AB - OBJECTIVES: To examine gender differences according to health status, social support, and DRG reimbursement in a population of elderly patients admitted to a hospital geriatric ward in Italy and also to examine the patterns of these differences across old age strata. DESIGN: Observational study PARTICIPANTS: A total of 2171 patients, 70 years and older (females = 1088), consecutively admitted for acute care to a geriatric ward in a general hospital during a 30 month period were included. Patients were stratified into 3 age groups: 70 to 79 (n = 1038, females = 521), 80 to 89 (n = 948, females = 476), and 90+ (n = 185, females = 91). MEASUREMENTS: Demographics, main reason for hospitalization, Charlson Index, APACHE II score, APACHE II-APS subscore, serum albumin, number of currently administered drugs, cognitive status, depression, functional status, length of stay, Diagnoses Related Group (DRG) weight, in-hospital, and 3-month mortality were recorded. Differences were evaluated according to gender across old age strata. RESULTS: In the group of 70- to 79-year-old patients, significant differences were found regarding number of comorbidities, biological and clinical markers of clinical severity (ie, serum albumin, APS, delirium), and functional status on admission (ie, the greater impairment was found in male patients, with a higher in-hospital and 3-month mortality). Moreover, females had less social support and more often live alone. DRG weight parallels clinical complexity, whereas length of stay is comparable. Gender differences were less evident in the 80-89 year-old patients and almost absent in those 90+. CONCLUSIONS: Data indicate that on hospital admission gender differences are significant in young old patients, but not in old and very old, suggesting a poor survivorship of males with increasing age. PMID- 19808154 TI - Perceived barriers to effective job performance among nursing assistants in long term care. AB - OBJECTIVES: This research explored perceived barriers to job performance among a national sample of nursing assistants (NAs). Specific objectives were (1) to clarify which of the problems identified by previous research are most troublesome for NAs, (2) to develop a reliable quantitative measure of perceived barriers to job performance, and (3) to test construct validity of the measure vis-a-vis work-related psychological empowerment and job satisfaction. METHODS: Nursing assistants attending the 2006 national conference of the National Association of Health Care Assistants completed a paper-and-pencil survey including 33 barriers to job performance and standardized measures of empowerment and job satisfaction. The barriers were also rated by a small sample of NAs at a single Georgia nursing home. RESULTS: Factor analysis of barriers items yielded a 30-item Nursing Assistants Barriers Scale (NABS) comprising 6 subscales: Teamwork, Exclusion, Respect, Workload, Work Stress, and New NAs. Lack of teamwork and exclusion from communication processes were rated as most problematic by both samples. The 6 NABS subscales were significantly and independently associated with empowerment and satisfaction; different barriers predicted the 2 constructs. DISCUSSION: This study is a first step toward quantitative assessment of NAs' perceptions of barriers to doing their jobs. Primary limitations are the select sample and use of a job satisfaction measure that may have artificially inflated correlations with the NABS. Nonetheless, results confirm the validity of the new scale as an operationalization of the barriers construct. CONCLUSION: The concept of barriers to job performance is a unique construct from work empowerment and satisfaction with one's job. Nursing assistants clearly differentiate various barriers, converging on workload and lack of teamwork as most problematic. Further work is needed to substantiate validity and reliability of the NABS, particularly with respect to NAs' actual job performance, intent to stay on the job versus leave, absenteeism, and turnover. PMID- 19808155 TI - The importance of standardized observations to evaluate nutritional care quality in the survey process. AB - OBJECTIVE: Guidelines written for government surveyors who assess nursing home (NH) compliance with federal standards contain instructions to observe the quality of mealtime assistance. However, these instructions are vague and no protocol is provided for surveyors to record observational data. This study compared government survey staff observations of mealtime assistance quality to observations by research staff using a standardized protocol that met basic standards for accurate behavioral measurement. Survey staff used either the observation instructions in the standard survey process or those written for the revised Quality Improvement Survey (QIS). METHODS: Trained research staff observed mealtime care in 20 NHs in 5 states during the same time period that survey staff evaluated care in the same facilities, although it could not be determined if survey and research staff observed the same residents during the same meals. Ten NHs were evaluated by government surveyors using the QIS survey instructions and 10 NHs were evaluated by surveyors using the standard survey instructions. RESULTS: Research staff observations using a standardized observation protocol identified a higher proportion of residents receiving inadequate feeding assistance during meals relative to survey staff using either the standard or QIS survey instructions. For example, more than 50% of the residents who ate less than half of their meals based on research staff observation were not offered an alternative to the served meal, and the lack of alternatives, or meal substitutions, was common in all 20 NHs. In comparison, the QIS survey teams documented only 2 instances when meal substitutes were not offered in 10 NHs and the standard survey teams documented no instances in 10 NHs. CONCLUSIONS: Standardized mealtime observations by research staff revealed feeding assistance care quality issues in all 20 study NHs. Surveyors following the instructions in either the standard or revised QIS surveys did not detect most of these care quality issues. Survey staff instructions for observation of nutritional care are not clearly written; thus, these instructions do not permit accurate behavioral measurement. These instructions should be revised in consideration of basic principles that guide accurate behavioral measurement and shared with NH providers to enable them to effectively implement quality improvement programs. PMID- 19808156 TI - Using video images to improve the accuracy of surrogate decision-making: a randomized controlled trial. AB - INTRODUCTION: When patients are unable to make important end-of-life decisions, doctors ask surrogate decision makers to provide insight into patients' preferences. Unfortunately, multiple studies have shown that surrogates' knowledge of patient preferences is poor. We hypothesized that a video decision tool would improve concordance between patients and their surrogates for end-of life preferences. OBJECTIVE: To compare the concordance of preferences among elderly patients and their surrogates listening to only a verbal description of advanced dementia or viewing a video decision support tool of the disease after hearing the verbal description. METHODS: This was a randomized controlled trial of a convenience sample of community-dwelling elderly subjects (> or = 65 years) and their surrogates, and was conducted at 2 geriatric clinics affiliated with 2 academic medical centers in Boston. The study was conducted between September 1, 2007, and May 30, 2008. Random assignment of patient and surrogate dyads was to either a verbal narrative or a video decision support tool after the verbal narrative. End points were goals of care chosen by the patient and predicted goals of care by the surrogate. Goals of care included life-prolonging care (CPR, mechanical ventilation), limited care (hospitalization, antibiotics, but not CPR), and comfort care (only treatment to relieve symptoms). The primary outcome measure was the concordance rate of preferences between patients and their surrogates. RESULTS: A total of 14 pairs of patients and their surrogates were randomized to verbal narrative (n=6) or video after verbal narrative (n=8). Among the 6 patients receiving only the verbal narrative, 3 (50%) preferred comfort care, 1 (17%) chose limited care, and 2 (33%) desired life-prolonging care. Among the surrogates for these patients, only 2 correctly chose what their loved one would want if in a state of advanced dementia, yielding a concordance rate of 33%. Among the 8 patients receiving the video decision support tool, all 8 chose comfort care. Among the surrogates for these patients, all 8 correctly chose what their loved one would want if in a state of advanced dementia, yielding a concordance rate of 100%. CONCLUSION: Patients and surrogates viewing a video decision support tool for advanced dementia are more likely to concur about the patient's end-of-life preferences than when solely listening to a verbal description of the disease. PMID- 19808157 TI - Religiousness, religious coping, and psychological well-being in nursing home residents. AB - OBJECTIVES: To measure the importance of religion among nursing home residents, describe their use of religious coping strategies, and examine the association between religiousness, religious coping, and psychological well-being. DESIGN: Cross-sectional study. SETTING: Two nursing facilities in Boston, Massachusetts. PARTICIPANTS: One hundred forty cognitively intact to moderately impaired long stay nursing home residents. MEASUREMENTS: Subjects rated religion as either "not important," "somewhat important," or "very important." Use of religious coping strategies was measured using the 14-item Brief RCOPE. The outcome measure, psychological well-being, was measured with the Bradburn Affect Balance Scale. Covariates included demographic variables and a measure of social engagement, comorbidity, functional status, and mental status. Linear regression was used to examine the association between religious importance and psychological well-being after adjusting for covariates. RESULTS: Subjects rated the importance of religion as follows: very important (54%), somewhat important (27%), and not important (19%). The mean score on the Affect Balance Scale was 5.9 +/- 2.1 (SD) (range 2-9). After multivariate adjustment, viewing religion as somewhat or very important (versus not important, P=.0019) and absence of negative religious coping strategies (P=.0083) were associated with better psychological well-being (with higher scores on the Affect Balance Scale) (P=.007). CONCLUSION: Religion was important to most older residents living in 2 religiously affiliated long term care facilities. Residents for whom religion was somewhat or very important and who did not use negative religious coping strategies are more likely to have better psychological well-being. PMID- 19808158 TI - The physician's role in long-term care culture change. AB - This article describes an approach to nursing home physician care that allows physicians to play a more integral role in the long-term care culture change movement. Through a simple refocusing of the traditional clinical assessment and plan, the physician's role is shaped and redefined, support staff more clearly see how the physician may contribute to the effort, and residents and families are reassured that the entire care team is working together to improve the care provided. PMID- 19808159 TI - Diabetes management in long-term care facilities: a practical guide. PMID- 19808160 TI - Does prematurity affect platelet indices? AB - PURPOSE: The current study objective was to compare blood platelet indices in preterm newborns (PTN) and full term newborns (FTN). MATERIALS AND METHODS: We introduced to our study 51 PTN (25 females, 26 males) and 55 FTN (25 females, 30 males). Platelet indices were estimated in blood samples collected from the umbilical artery. RESULTS: PTN demonstrated a decreased count of blood platelets (197 x 103/microL) as compared to FTN (287 x 103/microL), p=0.0001. Platelet hematocrit (PCT) also showed substantial differences in both groups (PTN=0.16% vs. FTN=0.22%; p=0.001). Mean platelet volume (MPV) was found to be nearly the same (PTN=8.02 fl, FTN=7.79 fl). Platelet distribution width (PDW) was higher in PTN (50.64%) than in FTN (46.54%), p=0.021. Large platelet count (LPLT) was diminished in PTN (5.23%) in comparison with FTN (6.12 %). CONCLUSIONS: A decreased count of blood platelets, platelet hematocrit and increased platelet distribution width may result from a low gestational age or a dysfunction of megakaryocytes and the placenta. Blood platelet indices may be vital in the diagnosis of haemostatic disorders. PMID- 19808161 TI - Predictive value of cavernosal peak systolic velocity in the flaccid penis. AB - PURPOSE: The routine use of intracavernosal injection before Doppler investigation may cause time loss, side effects due to vasoactive agents used and incorrect evaluation in the diagnosis of arterial insufficiency because of psychological inhibition and anxiety. Doppler investigation without intracavernosal injection avoids these pronounced disadvantages. In our study we tried to evaluate whether PSV values prior to intracavernosal injection are predictive in the diagnosis of arterial insufficiency in diabetic and non diabetic cases. MATERIAL AND METHODS: 120 male diabetic (type-2) and non-diabetic patients affected by erectile dysfunction (ED) were enrolled in this study. The Doppler parameters - PSV, end diastolic velocity (EDV) of both cavernosal arteries - were recorded before intracavernosal injection (ICI) (60 mg-2 ml papaverine hydrochloride (HCl)) and 5, 10 and 30 minutes after ICI into corpus cavernosum. Based on our results we chose 2 cut-off values for the pre-ICI PSV-10 cm/sec and 15 cm/sec. A PSV value less than 25 cm/sec after ICI was used as the diagnostic criteria for arterial insufficiency. We calculated the sensitivity, specificity, positive and negative predictive value of pre-ICI PSV in predicting arteriogenic impotence as diagnosed by post-ICI Doppler sonography. Only type 2 DM patients were included in our study. RESULTS: Statistically significant differences were found in the PSV values before and after ICI, ICI response and EDV values after ICI in both cavernous arteries when diabetic and nondiabetic groups compared. CONCLUSIONS: Flow in the cavernosal arteries in the flaccid state could determine nondiabetic patients with vasculogenic impotence with a high accuracy rate. PMID- 19808162 TI - Stem cell niche in the Drosophila ovary and testis; a valuable model of the intercellular signalling relationships. AB - One of the key factors determining the function of all types of stem cells is their location in a specific microenvironment called a niche which is understood as a system of adjacent cells directly influencing their ability to carry out self-renewal divisions. The cells which compose the niche influence cytophysiological processes of stem cells both directly via the intercellular junction system and via the synthesis and release of many protein regulatory substances which are ligands of specific receptors in a particular stem cell. These proteins are often the products of distinct genes whose expression tends to be specific for niche-composing cells. The niches formed of a few cells only observed in Drosophila gonads may become a valuable functional model in the studies of mammal stem cells since their analysis proves that the preservation of the stem cells' unique features does not require a large number of cells to be present in its vicinity. PMID- 19808163 TI - Cancer of the accessory breast--a case report. AB - Breast neoplasm may develop in ectopically located glandular tissue. This paper presents an interesting and rare case of a 50-year-old female who despite regular mammography screening examination developed an invasive accessory breast cancer. Clinical examination revealed a 2 cm - tumour localized 4 cm below the left inframammary fold. The lesion was immobile, the skin and the atrophic nipple were retracted, the tumour infiltrated the thoracic wall. Oligobiopsy and additional examinations showed an invasive stage IIIB ductal breast cancer (Bloom II, G-2). The receptor status was: ER(+), PGR(+), HER2(-). The increased level of cancer antigen 15.3 was found. The patient was submitted to pre-operative chemotherapy. She also underwent surgery and subsequently post-operative chemotherapy and radiotherapy. On the basis of the presented case, it could be concluded that the accessory mammary glands are out of the image of screening breast examinations. Accessory breast cancer is usually diagnosed by clinical examination and ultrasonography. Preventive resection of accessory breast in women at high risk of developing breast cancer can be considered as the treatment of choice in most patients. PMID- 19808164 TI - Blindness following rupture of hepatic hydatid cyst: a case report. AB - A 19 year-old woman admitted to Emergency Department with hypotension, sudden loss of vision and acute abdominal pain. Ultrasound and computed tomography demonstrated an occipital infarct in brain and ruptured intraperitoneal cyst of hydatid liver disease. Urgent laparotomy was performed and it included aspiration of cyst contents, peritoneal washing and drainage. Her vision loss improved by 15 hours postoperatively but generalized seizures were started. Weakness in all extremities was present. Cranial MRI demonstrated ischemia in the areas of middle, posterior and anterior cerebral arteries. She was discharged from the hospital with severe neurological deficits (unable to walk, not able to eat herself). Neurological deficits were improved with physiotherapy after two years. There was no recurrence of hydatid cysts in the follow-up of three years. We assumed that anaphylaxis after intraperitoneal rupture of hydatid liver cyst resulted with hypotension and reduced cerebral perfusion, caused the acute vision loss and other neurological symptoms. This unusual presentation of intraperitoneal rupture should be kept in mind particularly in endemic areas of hydatid disease. PMID- 19808165 TI - Massive retroperitoneal adrenal hemorrhage secondary to lung cancer metastasis treated by adrenal artery embolization. AB - The development of adrenal hemorrhage from lung cancer metastasis is extremely rare. A 54-year-old woman with stage IV non-small-cell lung cancer and known bilateral adrenal metastasis developed severe right-sided abdominal pain while undergoing chemotherapy treatment. A computerized tomography scan of the abdomen with intravenous contrast demonstrated massive right-sided retroperitoneal hemorrhage. The bleeding vessel was identified as the right middle adrenal artery. The patient underwent successful transcatheter embolization (TCE) and occlusion of the right middle adrenal artery using metallic coils, with resolution of the abdominal pain and stabilization of her hemoglobin. TCE can be used to control hemorrhage arising from metastatic involvement of the adrenal gland. PMID- 19808188 TI - The role for surgery in stage III non-small-cell lung cancer: can we reliably select the right patients? PMID- 19808190 TI - Cooperative group research endeavors in small-cell lung cancer: current and future directions. AB - The International Lung Cancer Congress (ILCC), now in its ninth year, is a key forum for representatives of cooperative groups in North America, Europe, and Japan to discuss ongoing and planned clinical trials in lung cancer. Many of the significant strides in lung cancer treatment often originate from investigations designed within the cooperative group system and were a feature of the 2008 ILCC. Small-cell lung cancer (SCLC) represents 15% of all lung cancers diagnosed annually and is characterized by rapid growth kinetics, disseminated metastases, and development of chemotherapy resistance. Many questions remain regarding the optimal use of radiation therapy and approaches for enhancing the effects of chemotherapy to improve clinical outcomes. Herein, we explore and outline the scientific vision of each cooperative group's SCLC research portfolio, as presented at the 2008 ILCC. Highlights include an ongoing Intergroup phase III study exploring differing radiation therapy schemes for limited-stage SCLC and a Southwest Oncology Group 0124 trial establishing platinum/etoposide as the standard of care for untreated extensive-stage SCLC in North America. Continued research efforts sponsored by these groups will represent the future of SCLC diagnosis and management. PMID- 19808191 TI - Understanding prognostic gene expression signatures in lung cancer. AB - In non-small-cell lung cancer (NSCLC), molecular profiling of tumors has led to the identification of gene expression patterns that are associated with specific phenotypes and prognosis. Such correlations could identify early-stage patients who are at increased risk of disease recurrence and death after complete surgical resection and who might benefit from adjuvant therapy. Profiling may also identify aberrant molecular pathways that might lead to specific molecularly targeted therapies. The technology behind the capturing and correlating of molecular profiles with clinical and biologic endpoints have evolved rapidly since microarrays were first developed a decade ago. In this review, we discuss multiple methods that have been used to derive prognostic gene expression signatures in NSCLC. Despite the diversity in the approaches used, 3 main steps are followed. First, the expression levels of several hundred to tens of thousands of genes are quantified by microarray or quantitative polymerase chain reaction techniques; the data are then preprocessed, normalized, and possibly filtered. In the second step, expression data are combined and grouped by clustering, risk score generation, or other means, to generate a gene signature that correlates with a clinical outcome, usually survival. Finally, the signature is validated in datasets of independent cohorts. This review discusses the concepts and methodologies involved in these analytical steps, primarily to facilitate the understanding of reports on large dataset gene expression studies that focus on prognostic signatures in NSCLC. PMID- 19808192 TI - Quality indicators in cancer care: development and implementation for improved health outcomes in non-small-cell lung cancer. AB - Non-small-cell lung cancer (NSCLC) care is multidisciplinary and complex in nature. However, there are few quality indicators that are widely accepted by the physicians who treat lung cancer. Quality indicators developed by the American Society of Clinical Oncology and National Comprehensive Cancer Network exist for breast and colon cancer, but not yet for lung cancer. In this article we review the current state of quality indicators in oncology care in general and for NSCLC in particular. Proposed quality metrics focus on diagnosis and staging, timeliness of care, supportive care and patient satisfaction. PMID- 19808193 TI - Utilization of internet-based survivorship care plans by lung cancer survivors. AB - BACKGROUND: Persons living after lung cancer remain an important subset of the cancer survivor population who may be at risk for serious health consequences of lung cancer and its treatments. The Institute of Medicine recommends survivorship care plans for all cancer survivors. MATERIALS AND METHODS: A program for the creation of survivorship care plans, OncoLife, was made publicly accessible. Users responded to queries regarding demographics, diagnosis, and treatment and received comprehensive, individualized guidelines for future care addressing issues such as late toxicity, second malignancy, fertility, and tumor recurrence. Data were maintained anonymously with approval from the Institutional Review Board. RESULTS: From May 2007 to November 2008, 3343 individuals completed OncoLife surveys. Lung cancer survivors comprised 4% (n=142), with a median age at diagnosis of 57 years and median current age of 59 years. Lung cancer survivors were 50% male and 83% Caucasian. Chemotherapy was used in the treatment of 78% of patients with lung cancer, radiation was used in 58%, and surgery was used in 38%. Most lung cancer survivors (62%) reported receiving follow-up care from only an oncologist, while 27% reported being monitored by a primary care provider (PCP) and an oncologist, and 6% saw only a PCP. Only 11% reported receiving survivorship information at the conclusion of therapy. CONCLUSION: OncoLife represents the first Web-based program for the creation of survivorship care plans, and lung cancer survivors appear willing to use this type of tool. Most have received multimodality care, placing them at risk for treatment-related late effects. Most survivors do not report receiving routine care from a PCP, so attention to survivor healthcare needs by the oncology community is particularly important. PMID- 19808194 TI - Results of trimodality therapy in patients with stage IIIA (N2-bulky) and stage IIIB non-small-cell lung cancer. AB - BACKGROUND: The survival rates for stage IIIA and stage IIIB non-small-cell lung cancer (NSCLC) are extremely poor with single-treatment modalities such as radiation therapy or surgery. The purpose of this study is to assess tolerability, response, surgical resectability, and survival of chemotherapy followed by chemoradiation therapy, and then followed by surgery in patients with stage IIIA (N2-bulky) or stage IIIB NSCLC. PATIENTS AND METHODS: Forty-eight patients with stage IIIA (N2-bulky) or stage IIIB (T4 N1-2 M0) NSCLC received 2 cycles of chemotherapy with cisplatin, mitomycin, and vindesine, subsequent radiation therapy (45 Gy, twice-daily 1.5 Gy) with simultaneous low-dose cisplatin and vindesine, followed by surgery. RESULTS: Forty-five patients completed induction chemoradiation therapy. Thirty-three patients (68.8%) had clinical response to induction treatment. Thirty-nine patients underwent a thoracotomy, with a complete resection rate of 62.5% (30/48). The pathologic response rate was 60% (27/45), with complete pathologic response of 8 patients. The median survival time for the total group of 48 patients was 23 months, with 3 and 5-year survival rates of 41.7% and 31.8%, respectively. Multivariate analysis showed that complete resection and pathologic response in surgical specimens were independent predictors of survival (P=.048 and P=.022). CONCLUSION: Preoperative sequence of chemotherapy followed by concurrent chemoradiation therapy is an effective approach in patients with stage IIIA (N2 bulky) and IIIB (T4 N1-2 M0) NSCLC. The operation after induction chemoradiation therapy should be performed in carefully selected patients with surgically resectable diseases. The patients who achieved complete resection and with pathologic response of tumor can benefit from surgery following induction chemoradiation therapy. PMID- 19808195 TI - Computed tomography-guided percutaneous needle biopsy of pulmonary nodules: impact of nodule size on diagnostic accuracy. AB - PURPOSE: This study was undertaken to compare the diagnostic accuracy and complication rate of computed tomography (CT)-guided percutaneous lung biopsies of lung nodules1.5 cm in diameter. PATIENTS AND METHODS: A total of 139 patients (age range, 18-89 years; mean, 62.5 years) underwent CT guided percutaneous fine-needle aspiration biopsy or 20-gauge core biopsy using an automated biopsy gun. In 37 patients, the lung nodule measured 1.5 cm (mean, 2.8 cm). Diagnostic accuracy was determined by cytopathology results. Major and minor complications were documented. RESULTS: Overall diagnostic accuracy, pneumothorax rate, and thoracostomy tube insertion rates were 67.6%, 34.5%, and 5%, respectively. Of the 98 patients with malignancy, 77 patients (78.6%) had a definite diagnostic biopsy. Overall, nodules>1.5 cm were statistically more likely to result in a diagnostic specimen (73.5%) than nodules1.5 cm than in those1.5 cm. However, the diagnostic accuracy for malignancy is high in both groups, with a low risk of complications. PMID- 19808196 TI - Pemetrexed-induced pneumonitis: a case report. AB - Pemetrexed is a structurally novel antifolate agent approved in combination with cisplatin for the treatment of patients with malignant pleural mesothelioma who have unresectable disease and for the therapy of previously treated patients with locally advanced or metastatic non-small-cell lung cancer (NSCLC) as a single agent or in association with cisplatin as a first-line treatment in patients with nonsquamous histology. Herein, we report a case of pemetrexed-induced pneumonitis. Because pemetrexed is being prescribed with increasing frequency for NSCLC and mesothelioma, we believe that physicians should be aware of this rare but serious complication. PMID- 19808197 TI - Myasthenia gravis and autoimmune Addison disease in a patient with thymoma. AB - The association of thymoma with myasthenia gravis has been well documented. However, the relationship between these two syndromes and Addison disease are very rarely encountered in clinical practice. We report on a 32-year-old man who underwent a resection for thymoma 48 months ago. The diagnosis of Addison disease was made followed by a diagnosis of myasthenia gravis on the basis of a high titer of acetylcholine receptor levels. The treatment of oral prednisolone 7.5 mg/day and oral prostigmine 180 mg/day was initiated. His symptoms and physical signs were improved after this treatment. To our knowledge, this is the fourth reported case of thymoma synchronously associated with myasthenia gravis and Addison disease. PMID- 19808198 TI - MAGRIT: the largest-ever phase III lung cancer trial aims to establish a novel tumor-specific approach to therapy. AB - This clinical trial summary provides the background and rationale for a randomized trial, MAGRIT, to investigate the efficacy of MAGE-A3 antigen-specific cancer immunotherapeutic (ASCI) agents in preventing cancer relapse, when administered after tumor resection, in patients with MAGE-A3-positive stages IB, II, and IIIA non-small-cell lung cancer. The study will also evaluate potential side effects of MAGE-A3 ASCIs. The primary endpoint is disease-free survival. The secondary endpoint is prospective validation of the gene signature predictive of benefit from MAGE-A3 ASCI therapy. PMID- 19808201 TI - Why not support(ed) parenting? PMID- 19808202 TI - A public health approach to promoting better mental health outcomes for children of parents with a psychiatric disability. AB - TOPIC: This paper describes the development of a national initiative to promote better mental health outcomes for children of parents with a psychiatric disability. PURPOSE: The purpose of the paper is twofold. First, the paper reflects on the policy framework and factors that led to the development of the national initiative and its outcomes. Secondly, the paper is intended to provide information about strategies utilized in the initiative that may be transferable to other settings. SOURCES USED: Health and social policy literature and personal observations are the major sources used. CONCLUSIONS: National public health policy concepts of the promotion of positive health and well-being and the prevention of illness have been a valuable vehicle for increasing awareness of, and responses to, the mental health of Australian children, parents, and families where a parent has a psychiatric disability. The involvement of young people and families in the development of policies and service guidelines that impact upon their lives, in partnership with professionals, government and non-government organizations, warrants continued emphasis. Translating policy into practice requires much ongoing work that can be supported by coalition building, development of a sound research and evidence base, and effective workforce education. PMID- 19808203 TI - Adoption and safe families act state statutes regarding parents with mental illnesses: a review and targeted intervention. AB - TOPIC: This paper describes an intervention targeting states that list a parental mental illness/disability as an "aggravated circumstance" under the Adoption and Safe Families Act of 1997 (ASFA), resulting in reasonable efforts not required to reunify a family. PURPOSE: This paper delineates the results from our review of ASFA state statutes, the development of a model ASFA statute, and strategies to educate legislators and the public about the impact of discrimination that parents with mental illnesses encounter because of ASFA legislation with the intent of modifying state ASFA legislation. SOURCES USED: The following sources were used for this educational initiative: a literature review and a review of ASFA state statutes. CONCLUSIONS: Adoption of the model ASFA state statute is simply a first step in an effort to end the discrimination that parents with psychiatric disabilities face; additional efforts are also noted. PMID- 19808204 TI - Promoting collaborative practice for children of parents with mental illness and their families. AB - OBJECTIVE: This paper describes the implementation and outcomes of a trial interagency protocol to enhance the responsiveness of community services for children of parents with mental illness and their families. METHODS: Protocol implementation was supported by the allocation of designated project workers, Interagency and Advisory Committees, and cross-agency training. Survey data on staff perceptions of collaborative practice were obtained at baseline and 12 months later. Participants represented a range of child and adult-serving agencies, and included consumers and family members. RESULTS: Intra- and interagency collaboration increased, with staff reporting on barriers, e.g., work load and time pressures, and benefits, e.g., shared expertise. They indicated collaboration was of value to the children and families with whom they worked. CONCLUSIONS: Interagency efforts can enhance collaborative practice. Stakeholders continue to work together to refine their efforts and create new approaches to improving outcomes for all family members living with parental mental illness. PMID- 19808205 TI - Supporting clubhouse members in their role as parents: necessary conditions for policy and practice initiatives. AB - OBJECTIVE: Given that the majority of adults with mental illness are parents, it is likely that a substantial number of members in the Clubhouse community are parents. Supporting members in their role as parents presents meaningful, philosophical and practical challenges for both individual Clubhouses and the Clubhouse movement. Supporting parents within the Clubhouse, however, is a necessary and logical step, consistent with the Clubhouse emphasis on rehabilitation and recovery. The current study explored the conditions necessary for supporting members in the parenting role in an existing Clubhouse. METHODS: A grounded-theory, ethnographic approach was used to collect data from multiple stakeholders including Clubhouse members, staff, and Board of Directors. A total of nine focus groups with 30 participants were conducted. RESULTS: Five themes emerged reflecting the conditions necessary to support parents in the Clubhouse: 1) securing stakeholder buy-in; 2) identification of shared values and principles; 3) clarification about how supporting parents will affect current Clubhouse activities; 4) facilitation of ongoing communication about changes; and 5) exploration of opportunities to maximize resources to support all Clubhouse members. CONCLUSIONS: Findings confirm the philosophical desirability and practical feasibility of supporting members in the parenting role, and identify fundamental challenges to philosophy and practice raised by the paradigm shift from thinking about individuals to thinking about families. PMID- 19808206 TI - Family options for parents with mental illnesses: a developmental, mixed methods pilot study. AB - OBJECTIVE: The objective of this paper is to provide a description of Family Options, a rehabilitation intervention for parents with serious mental illnesses and their children focusing on recovery and resilience, and to report the findings from a pilot study at 6-months post-enrollment for participating mothers. METHODS: A developmental design, and mixed quantitative and qualitative methods facilitate an in-depth understanding of Family Options and its impact on parents early in the implementation process. RESULTS: Participating families faced significant challenges, including long-term mental health conditions in adults, and emotional and behavioral difficulties in children. Data from mothers (n = 22) demonstrate significant improvements in well-being, functioning, and supports and resources at 6 months post-enrollment in Family Options. Mothers report help from Family Options staff consistent with the intervention as conceptualized, and high levels of satisfaction with the intervention as delivered. CONCLUSIONS: Innovative study design and analytic strategies are required to build the evidence base and promote rapid dissemination of effective interventions. Findings from this study will assist purveyors in refining the intervention, and will lay the groundwork for further replication and testing to build the evidence base for parents with serious mental illnesses and their families. PMID- 19808207 TI - Improving outcomes for pregnant and parenting foster care youth with severe mental illness: an evaluation of a transitional living program. AB - OBJECTIVE: This study provides preliminary evidence for the effectiveness of a residential program with comprehensive wrap-around services for pregnant and parenting foster care youth with severe mental illness or severe emotional disturbance who are preparing to transition to independent living. METHODS: Twenty-five females ages 18 to 21 participated. Program effectiveness was determined by tracking monthly status changes, such as education, employment, and hospitalizations, as well as by comparing scores on four bi-annually administered standardized assessments of parenting competency and stress, child maltreatment risk, and mental health symptoms. RESULTS: Assessment findings indicate that program participation is associated with positive changes in participants' familial relationships, family responsibility and care, proper parenting behavior and feelings, and parental distress and competency, but no change in mental health symptoms. Positive behavior changes associated with program participation were observed in education, employment, and low numbers of suspected and substantiated child maltreatment reports. Negative behavior changes associated with the program were frequency of AWOL incidents and subsequent pregnancies. CONCLUSIONS: This is the first study to evaluate a program designed for this very unique population and has implications for program design and policy. Future research with more rigorous experimental design is necessary in order to understand the critical ingredients of programs serving this vulnerable population. PMID- 19808208 TI - A "snapshot" of Australian programs to support children and adolescents whose parents have a mental illness. AB - OBJECTIVE: The aim of this study was to provide a "snapshot" of Australian programs targeting children and adolescents whose parents have a mental illness (COPMI). Specifically we aimed to (i) identify the aims and strategies of COPMI programs, (ii) ascertain the theoretical basis of these programs, (iii) determine the level and quality of evaluation protocols implemented, and (iv) draw on the practice wisdom of program facilitators for future program development. METHODS: Eighteen program facilitators responsible for 26 programs were identified through COPMI websites, e-discussion lists and snowball recruiting and invited to participate in a one hour interview based on the above four aims. Transcripts were qualitatively analyzed. RESULTS: Many COPMI programs target children aged between 8-13 who exhibit no psychological problems and live in urban areas. Overall, program facilitators aim to create opportunities for peer interaction, enhance coping, self-esteem, understanding of mental illness and offer respite from caring. Interventions are primarily supportive and preventative. While most programs are evaluated, measures on the whole are not standardized and/or gauge children's satisfaction of the program. CONCLUSION: Program facilitators require greater support, resources and training when evaluating COPMI programs. Program goals provide an indication of the evaluation measures required. PMID- 19808209 TI - Early outcomes of a pilot psychoeducation group intervention for children of a parent with a psychiatric illness. AB - OBJECTIVE: This study reports early findings of a still-developing Youth Education and Support (YES) pilot intervention of multifamily group psychoeducation for youth with a parent with a psychiatric illness. METHODS: Hypotheses predicted YES participants would increase pre-to-post: 1) Knowledge of psychiatric illness and recovery, and 2) Coping. A purposive sample of 17 youth, ages 10-16, participated in six lively, activity-focused, two-hour sessions within four groups facilitated by the PI and professionals employed within two public mental health agencies. Data revealed a significant increase in pre-to post youth-reported knowledge (p= less than .001) and no significant change in overall coping. At post-intervention, youth reported significantly increased use of the coping skills of avoiding problems and relaxing, as indicated within these coping subscales. CONCLUSIONS: Study limitations merit interpretation caution. They are useful for future research, including development and testing of youth psychoeducation programs with longer interventions, more emphases on coping, parent-inclusion, and larger samples using randomized, experimental designs. Suggestions for research, practice, and policy are provided. PMID- 19808210 TI - Termination of parental rights proceedings: legal considerations and practical strategies for parents with psychiatric disabilities and the practitioners who serve them. AB - TOPIC: This article examines the issue of parenting with a psychiatric disability and the risk of custody loss for these parents. PURPOSE: The purpose of this article is to examine the parental rights of parents with psychiatric disabilities and the use of the Americans with Disabilities Act as a legal defense. The current status of the law on terminating parental rights for people with mental illnesses is reviewed and the use of the ADA as a defense is described. Other defense strategies for parents and practitioners are also addressed. SOURCES USED: Sources for this paper include published literature and research on issues related to parenting with mental illnesses. Pertinent American case law and federal statutes were also examined. CONCLUSION: While using the Americans with Disabilities Act as a defense has had limited success in state termination of parental rights proceedings, some courts have declared it applicable in certain instances. Legal strategies such as raising this issue early in the termination process and practical strategies such as having a child care emergency plan are crucial to parents and practitioners who are navigating the child welfare system. PMID- 19808211 TI - Second chance at motherhood. PMID- 19808212 TI - My journey as an advocate for children of parents with a mental illness. PMID- 19808213 TI - The support for evidence-based training strategies. PMID- 19808215 TI - A randomized controlled study of weekly and biweekly dosing of epoetin alfa in CKD Patients with anemia. AB - BACKGROUND AND OBJECTIVES: In clinical practice, physicians often use once-weekly (QW) and biweekly (Q2W) dosing of epoetin alfa to treat anemia in patients with chronic kidney disease (CKD). Although the literature supports this practice, previous studies were limited by short treatment duration, lack of randomization, or absence of the approved three times per week (TIW) dosing arm. This randomized trial evaluated extended dosing regimens of epoetin alfa, comparing QW and Q2W to TIW dosing in anemic CKD subjects. The primary objective was to show that treatment with epoetin alfa at QW and Q2W intervals was not inferior to TIW dosing. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: 375 subjects with stage 3 to 4 CKD were randomized equally to the three groups and treated for 44 wk; to explore the impact of changing from TIW to QW administration on hemoglobin (Hb) control and adverse events, subjects on TIW switched to QW after 22 wk. The Hb was measured weekly, and the dose of epoetin alfa was adjusted to achieve and maintain an Hb level of 11.0 to 11.9 g/dl. RESULTS: Both the QW and Q2W regimens met the primary efficacy endpoint. More subjects in the TIW group than in the QW and Q2W groups exceeded the Hb ceiling. Adverse events were similar across treatment groups and consistent with the morbidities of CKD patients. CONCLUSIONS: Administration of epoetin alfa at QW and Q2W intervals are potential alternatives to TIW dosing for the treatment of anemia in stage 3 to 4 CKD subjects. PMID- 19808216 TI - Bone disease in medullary sponge kidney and effect of potassium citrate treatment. AB - BACKGROUND AND OBJECTIVES: In medullary sponge kidney (MSK)-a common malformative renal condition in patients with calcium nephrolithiasis-hypercalciuria, incomplete distal renal tubular acidosis, and hypocitraturia are common. Clinical conditions with concomitant hypercalciuria and/or incomplete distal renal tubular acidosis are almost invariably associated with bone disease, making osteopathy highly likely in MSK, too. Patients with MSK have never been investigated for osteopathy; neither has the potential effect of potassium citrate administration (CA) on their urinary metabolic risk factors and on bone mineralization. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: These issues were retrospectively analyzed in 75 patients with MSK and primary stone risk factor (PSRF; hypercalciuria, hypocitraturia, hyperuricosuria, and/or hyperoxaluria) on an outpatient basis; 65 received CA (2.9 +/- 0.8 g/d), whereas 10 received only general "stone clinic" suggestions. The 24-h urinary excretion of calcium, phosphate, oxalate, uric acid, and citrate; morning urine pH; serum biochemistry; and bone mineral density were investigated at baseline and at the end of follow-up (78 +/- 13 and 72 +/- 15 mo in groups A and B, respectively). RESULTS: CA led to a significant rise in urinary pH and citrate and decreased urinary calcium and phosphate (all P < 0.001). Patients with MSK and PSRF had reduced bone density. Bone density improved significantly in the group that was treated with oral CA. CONCLUSIONS: Bone disease is very frequent in patients with MSK and concomitant PSRF. Long term CA improves bone density. The concurrent effects of treatment on PSRF suggest that the subtle acidosis plays a pivotal role in bone disease and hypercalciuria in patients with MSK. PMID- 19808218 TI - More mixed messages in terms of salt. PMID- 19808217 TI - Longitudinal formulas to estimate GFR in children with CKD. AB - BACKGROUND AND OBJECTIVES: Whereas current GFR estimating equations approximate direct GFR measurement at a single time point, formulas that capitalize on changes in easily measured biologic parameters could improve the accuracy and precision of GFR estimation. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: In the Chronic Kidney Disease in Children Cohort (aged 1 to 16 yr), we measured GFR by plasma disappearance of iohexol (iGFR) and biomarkers in the first two annual visits. Models took the form GFR(2) = a[GFR(1)/40](b)[X(2)/X(1)](c), where GFR(2) and GFR(1) represented the current and previous years' iGFR, 40 ml/min per 1.73 m(2) was the cohort mean, and X(2)/X(1) was the change in predictors over time. Using data from 360 participants with a median age of 12.1 yr, we evaluated the predictive performance of a past GFR measurement and 20 other variables using a two-thirds random sample of the data. A one-third sample was reserved for validation. RESULTS: Previous iGFR measurements were strongly predictive of subsequent iGFR and adding change in height/serum creatinine significantly improved the explanatory power to 78%. In the validation set, the correlation between estimated and measured GFR was 0.88, and 48 and 88% of estimated GFRs were within 10 and 30% of observed iGFRs. When the past GFR measurement was not used, addition of change in markers to a cross-sectional model did not improve prediction. CONCLUSIONS: Longitudinal formulas to estimate iGFR capitalize on the high predictive power of previous iGFR measurements and in this study yielded a parsimonious prediction model with the potential for assessing progression in the clinical setting. PMID- 19808219 TI - The changing phenotype of academic nephrology -- a future at risk? AB - BACKGROUND AND OBJECTIVES: Academic nephrology faces increasing challenges in faculty hiring and development. However, it is unknown how these pressures have affected the number and demographics of academic nephrologists. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Using the Association of American Medical Colleges Faculty Roster database, changes were analyzed in MD nephrology, as well as other internal medicine subspecialty, faculty from 1998 to 2008. RESULTS: There were 1315 full-time MD nephrology faculty in 2008; this fell by 4.9% over the past decade. There were fewer junior, and more senior, faculty over this period. This was associated with 12.4% fewer tenured, 22.3% fewer tenure track, and an 11.5% increase in nontenure track, academic nephrologists. Academic nephrologists who are U.S. medical school graduates declined by 11.9%, while those who were international medical school graduates increased by 13.2%; nephrology has a greater percentage of international medical school graduates than any other internal medicine subspecialty. Female nephrology faculty increased by 14.3%, while male faculty fell by 9.5%. Asian nephrology faculty increased by 41.3%, while Caucasians declined by 15.2%. Similar changes in all the above parameters were seen for most other internal medicine subspecialties. The nephrology research programs at the top 20 research institutions, as compared with all other nephrology programs, had a greater decline in total MD, male, tenure track, and junior faculty. CONCLUSIONS: These data suggest that the future of academic nephrology is at risk. The decline in nephrology faculty provides incentive for leaders in academic nephrology to improve recruiting and retention practices. PMID- 19808221 TI - Predictors of complication after percutaneous ultrasound-guided kidney biopsy in HIV-infected individuals: possible role of hepatitis C and HIV co-infection. AB - BACKGROUND AND OBJECTIVES: HIV-infected patients often undergo kidney biopsy. The risks of percutaneous ultrasound-guided kidney biopsy in this population are not well established. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This was a case control, single-center study of 1116 (243 with HIV infection and 873 without) consecutive ultrasound-guided biopsies from 1024 patients. The primary outcome was any major or minor complication. Major complications included biopsy associated bleeding that required transfusion, angiography, or surgery; hypotension that required intervention; and death. Minor complications included development of a hematoma or gross hematuria. The odds of complication was assessed with logistic regression. RESULTS: Overall complication rates (8.6 versus 7.2%) did not significantly differ between HIV-infected and noninfected individuals. HIV-positive status did not predict complication. In the entire cohort, hepatitis C infection was associated with a 2.08 (95% confidence interval [CI] 1.47 to 2.93) increased odds of complication, and each 10,000-cells/mm(3) decrease in prebiopsy platelet count a 1.05 (95% CI 1.02 to 1.08) increased odds of complication. In addition, prebiopsy hematocrit <30% and estimated GFR <30 ml/min per 1.73 m(2) were associated with major complication. Whereas the association of prebiopsy platelet count was not modified by HIV infection, hepatitis C/HIV co-infection was associated with a 5.71 (95% CI 1.89 to 17.2) increased odds of complication as compared with 1.27 (95% CI 0.73 to 2.19) in hepatitis C-positive/HIV-negative individuals. CONCLUSIONS: Ultrasound-guided percutaneous kidney biopsy is a relatively safe, well-tolerated procedure in the HIV-infected population. HIV-infected individuals who are co-infected with hepatitis C seem to be at greatest risk. PMID- 19808220 TI - Hemodialysis-induced repetitive myocardial injury results in global and segmental reduction in systolic cardiac function. AB - BACKGROUND AND OBJECTIVES: Hemodialysis (HD)-induced regional wall motion abnormalities (RWMAs) are common in HD patients and driven by ischemia. In nondialysis patients, repeated ischemia leads to chronic reduction in left ventricular (LV) function. HD-induced myocardial ischemia may initiate the same process. We examined the effect of HD-induced repetitive myocardial stunning on global and regional LV function. DESIGN, SETTING, PARTICIPANTS & MEASUREMENTS: We analyzed data from 30 patients, previously identified as developing HD-induced myocardial ischemia. Serial echocardiographic assessments of global and regional LV performance were performed at baseline and repeated after 12 mo. RESULTS: Several patients developed segments with a fixed reduction in systolic function of >60% after 1 yr. In this patient group, there was a significant reduction in resting LV ejection fraction (EF) from 61.5 +/- 10.1% to 52.9 +/- 8.6% (P < 0.007). Peak LV EF in response to dialysis also decreased from 59.5 +/- 10% versus 49.9 +/- 6.5% (P < 0.003), with a consequent increase in HD-induced hypotension (P < 0.0001). CONCLUSIONS: HD-induced myocardial stunning may progress over 12 mo to the development of regional fixed systolic dysfunction, consistent with underlying myocardial hibernation and fibrosis. This may be an important and potentially modifiable process in the development of heart failure in HD patients. PMID- 19808222 TI - Eosinophil count is positively correlated with albumin excretion rate in men with type 2 diabetes. AB - BACKGROUND AND OBJECTIVES: Patients with allergic disorders such as allergic rhinitis or asthma have been reported to be at increased risk for atherosclerosis. In this study, we evaluated the relationships between peripheral eosinophil count and degree of albumin excretion rate, which is a useful marker of cardiovascular mortality as well as diabetic nephropathy in patients with type 2 diabetes. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We evaluated relationships of peripheral eosinophil count to degree of albumin excretion rate as well as to major cardiovascular risk factors, including age, BP, serum lipid concentration, and glycemic control (glycosylated hemoglobin); body mass index; current treatment for diabetes; smoking status; and presence of cardiovascular disease in 783 patients (416 men and 367 women) with type 2 diabetes. RESULTS: Log(eosinophil count) was positively associated with systolic BP (r = 0.124, P = 0.0108), serum triglyceride concentration (r = 0.108, P = 0.0284), and log(albumin excretion rate) (r = 0.301, P < 0.0001) in men; however, no association was found between log(eosinophil count) and log(albumin excretion rate) (r = 0.085, P = 0.1050) in women. Multivariate linear regression analysis demonstrated that log(eosinophil count) (beta = 0.260, P < 0.0001), duration of diabetes (beta = 0.203, P = 0.0003), glycosylated hemoglobin (beta = 0.117, P = 0.0238), systolic BP (beta = 0.205, P = 0.0001), and serum triglyceride concentration (beta = 0.162, P = 0.0038) were independent determinants of log(albumin excretion rate) in men. CONCLUSIONS: Allergic disorders may be associated with microalbuminuria in men with type 2 diabetes. PMID- 19808223 TI - The journey from vitamin D-resistant rickets to the regulation of renal phosphate transport. AB - In 1937, Fuller Albright first described two rare genetic disorders: Vitamin D resistant rickets and polyostotic fibrous dysplasia, now respectively known as X linked hypophosphatemic rickets (XLH) and the McCune-Albright syndrome. Albright carefully characterized and meticulously analyzed one patient, W.M., with vitamin D-resistant rickets. Albright subsequently reported additional carefully performed balance studies on W.M. In this review, which evaluates the journey from the initial description of vitamin D-resistant rickets (XLH) to the regulation of renal phosphate transport, we (1) trace the timeline of important discoveries in unraveling the pathophysiology of XLH, (2) cite the recognized abnormalities in mineral metabolism in XLH, (3) evaluate factors that may affect parathyroid hormone values in XLH, (4) assess the potential interactions between the phosphate-regulating gene with homology to endopeptidase on the X chromosome and fibroblast growth factor 23 (FGF23) and their resultant effects on renal phosphate transport and vitamin D metabolism, (5) analyze the complex interplay between FGF23 and the factors that regulate FGF23, and (6) discuss the genetic and acquired disorders of hypophosphatemia and hyperphosphatemia in which FGF23 plays a role. Although Albright could not measure parathyroid hormone, he concluded on the basis of his studies that showed calcemic resistance to parathyroid extract in W.M. that hyperparathyroidism was present. Using a conceptual approach, we suggest that a defect in the skeletal response to parathyroid hormone contributes to hyperparathyroidism in XLH. Finally, at the end of the review, abnormalities in renal phosphate transport that are sometimes found in patients with polyostotic fibrous dysplasia are discussed. PMID- 19808224 TI - Randomized controlled trial of icodextrin versus glucose containing peritoneal dialysis fluid. AB - BACKGROUND AND OBJECTIVES: While peritoneal dialysis with icodextrin is commonly used in patients with poor peritoneal membrane characteristics, the data on the usefulness of this solution in patients with lower transport characteristics are limited. The study was designed to compare icodextrin to glucose in Chinese prevalent peritoneal dialysis patients of different peritoneal transport characteristics (PET) categories. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This was a randomized, double-blind, perspective control study. Stable prevalent continuous ambulatory peritoneal dialysis (CAPD) patients were randomized to either 7.5% icodextrin (ICO) or 2.5% glucose (GLU) solution for 4 wk. Peritoneal membrane function was measured to define PET category in baseline. Creatinine clearance (Ccr), urea nitrogen clearance (C(BUN)), ultrafiltration (UF) during the long night dwell, dialysate, and metabolic biomarkers were measured at baseline, 2, and 4 wk. UF, Ccr, and C(BUN) were compared among different PET categories. RESULTS: A total of 201 CAPD patients were enrolled in the study. There were no baseline differences between the groups. Following 2 and 4 wk of therapy, Ccr, C(BUN,) and UF were all significantly higher in the ICO versus the GLU group. Additionally, switching to ICO resulted in a significant increase in UF in high, high-average, and low-average transporters as compared with baseline. The extent of increased UF was more obvious in higher transporters. Blood cholesterol level in the ICO group decreased significantly than that in the GLU group. CONCLUSION: Compared with glucose-based solution, 7.5% icodextrin significantly improved UF and small solute clearance, even in patients with low average peritoneal transport. PMID- 19808225 TI - Interstitial cystitis: an unsolved enigma. AB - Painful bladder syndrome/interstitial cystitis (PBS/IC) is a chronic disease of unknown etiology characterized by vague bladder pain and nonspecific urinary symptoms, such as urgency and frequency. Although it was initially considered to be a rare condition, its prevalence has significantly increased, possibly because of different definitions used and greater physician awareness. Because of the multiple diagnostic criteria used, there is significant variation in its prevalence. In addition, there is often a delay in the diagnosis of PBS/IC. It affects predominantly women of middle age, and it significantly decreases quality of life. Diagnosis of PBS/IC is mainly a diagnosis of exclusion; there are no characteristic symptoms or pathognomonic findings. Therefore, it is important to rule out diseases that have a similar clinical picture (i.e., urinary infections, bladder carcinoma) but definite therapies and worse prognosis if left untreated. PBS/IC management suffers from lack of evidence; many therapies are empiric or based on small studies and case series. Treatment includes supportive therapies (psychosocial, behavioral, physical), oral treatments, and intravesical treatments, whereas other more invasive treatments such as electric neuromodulation and reconstructive surgery are reserved for refractory cases. Physicians should always keep in mind the diagnosis of PBS/IC in patients presenting with chronic urinary symptoms after excluding other more common diseases. PMID- 19808226 TI - Correlates and outcomes of fatigue among incident dialysis patients. AB - BACKGROUND & OBJECTIVES: Fatigue is a debilitating symptom experienced by patients undergoing dialysis, but there is only limited information on its prevalence and its association with patient outcomes. This study examines the correlates of self-reported fatigue at initiation of dialysis and after 1 yr and assesses the extent to which fatigue was associated with health-related quality of life and survival. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: A longitudinal cohort of 917 incident hemodialysis and peritoneal dialysis patients who completed the CHOICE Health Experience Questionnaire (CHEQ) participated in the study. Fatigue was assessed using the SF-36 vitality scale. Known predictors of fatigue including sociodemographic and psychosocial factors, dialysis-related factors, biochemical variables including inflammatory markers, comorbidities, and medications were used as covariates. RESULTS: A low vitality score was independently associated with white race, higher Index of Coexistent Disease score, higher body mass index, lack of physical exercise, antidepressant use, and higher C-reactive protein levels (CRP). A lower vitality score was strongly associated with lower SF-36 physical functioning, mental health, bodily pain scores, and decreased sleep quality (all P < 0.001) at baseline. Among surviving participants, higher serum creatinine at baseline was associated with preserved vitality at 1 yr. Patients with the highest baseline vitality scores were associated with longer survival (hazard ratio 0.75; 95% CI 0.58 to 0.96, P = 0.03). CONCLUSIONS: The findings of this study demonstrate that ESRD patients experience profound levels of fatigue and elucidate its correlates. Also, the association of fatigue with survival may have significant implications for this population. PMID- 19808227 TI - Renal function in glycogen storage disease type I, natural course, and renopreservative effects of ACE inhibition. AB - BACKGROUND AND OBJECTIVES: Renal failure is a major complication in glycogen storage disease type I (GSD I). We studied the natural course of renal function in GSD I patients. We studied differences between patients in optimal and nonoptimal metabolic control and possible renoprotective effects of angiotensin converting enzyme inhibition. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Thirty-nine GSD I patients that visited our clinic were studied. GFR and effective renal plasma flow (ERPF) were measured by means of I(125) iothalamate and I(131) hippuran clearance and corrected for body surface area. Microalbuminuria was defined as >2.5 mg albumin/mmol creatinine and proteinuria as >0.2 g protein per liter. Optimal metabolic control was present when blood glucoses were >3.5 mmol/L, urine lactate/creatinine ratios <0.06 mmol/mmol, triglycerides <6.0 mmol/L, and uric acid concentrations <450 micromol/L. RESULTS: Quadratic regression analysis showed a biphasic pattern in the course of GFR and ERPF related to age. Microalbuminuria was observed significantly less frequently in the patients with optimal metabolic control compared with the patients with nonoptimal metabolic control. A significant decrease in GFR was observed after starting ACE inhibition. CONCLUSIONS: This study describes a biphasic pattern of the natural course of GFR and ERPF in GSD I patients, followed by the development of microalbuminuria and proteinuria. Optimal metabolic control has a renoprotective effect on the development of microalbuminuria and proteinuria in GSD I patients. Treatment with ACE inhibitors significantly decreases the GFR, especially in GSD I patients with glomerular hyperfiltration. PMID- 19808228 TI - Combining near-subject absolute and relative measures of longitudinal hydration in hemodialysis. AB - BACKGROUND AND OBJECTIVES: The feasibility and additional value of combining bioimpedance analysis (BIA) with near-subject absolute measurement of total body water using deuterium dilution (TBW(D)) in determining longitudinal fluid status was investigated. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Fifty-nine hemodialysis patients (17 female; age 58.4 +/- 16.1 yr; body mass index 27.0 +/- 5.4) were enrolled into a 12-mo, two-center, prospective cohort study. Deuterium concentration was measured in breath by flowing-afterglow mass spectrometry using a validated protocol ensuring full equilibration with the TBW; BIA was measured using a multifrequency, multisegmental device. Comorbidity was quantified by the Stoke score. Clinicians were blinded to body composition data. RESULTS: At baseline and 12 mo, there was an incremental discrepancy between TBW(BIA) and TBW(D) volumes such that greater comorbidity was associated with increasing overhydration. Forty-three patients who completed the study had no longitudinal differences in the prescribed or achieved postdialysis weights. In contrast, TBW(D) increased without a change in TBW(BIA) (mean difference -0.10 L). Changes in TBW and lean body mass differed according to baseline comorbidity; without comorbidity, BIA also identified an increase in TBW and lean body mass, whereas with increasing comorbid burden, BIA failed to demonstrate increases in tissue hydration identified by TBW(D). CONCLUSIONS: Combined near-patient measurements of absolute and BIA-estimated TBW are achievable in a dialysis facility by identifying changes in body composition not fully appreciated by routine assessment. BIA underestimates tissue overhydration that is associated with comorbidity, resulting in reduced sensitivity to longitudinal increases during a 12-mo period. PMID- 19808229 TI - The alphabet soup of kidney transplantation: SCD, DCD, ECD--fundamentals for the practicing nephrologist. AB - There is significant variability in the quality of deceased-donor kidneys that are used for transplantation. The quality of the donor kidney has a direct effect on important clinical outcomes such as acute rejection, delayed graft function, and patient and allograft survival. Expanded-criteria donors (ECDs) refer to older kidney donors (> or =60 yr) or donors who are aged 50 to 59 yr and have two of the following three features: Hypertension, terminal serum creatinine >1.5 mg/dl, or death from cerebrovascular accident. By definition, ECD kidneys have a 70% greater likelihood of failure compared with one from a 35-yr-old male donor who died from a motor vehicle accident. Donation after cardiac death (DCD) is a small but rapidly growing fraction of donors. An ECD kidney transplant recipient has a projected average added-life-years of 5.1 yr compared with 10 yr for a kidney recipient from a standard-criteria donor. Kidney transplantation from DCD seems to have similar allograft and patient survival compared with kidney from donation after brain death; however DCD transplantation has a 42 to 51% risk for delayed graft function (need for at least one dialysis treatment during the first week after transplantation) compared with 24% in an standard-criteria donor kidney transplant. Familiarity with the comprehensive allocation rules governing different categories of deceased-donor kidneys by the nephrologists and dialysis team providers is essential to maximizing patient autonomy and to improve the outcomes of kidney transplantation. PMID- 19808230 TI - Dystroglycan in the diagnosis of FSGS. AB - BACKGROUND AND OBJECTIVES: alpha- and beta-dystroglycan (DG), which link the actin cytoskeleton of the podocyte to the glomerular basement membrane, are maintained in FSGS but decreased in minimal change disease (MCD). Fibrosis has been linked to increased fibroblast-specific protein-1 (FSP1) and epithelial mesenchymal transition. We studied DG, FSP1, and podocyte differentiation in FSGS variants and cases of suspected FSGS. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We studied renal biopsies with FSGS, not otherwise specified (NOS), tip lesion, or collapsing variants (COLL), versus secondary FSGS or cases without segmental sclerotic lesions where a diagnosis of MCD versus FSGS could not be established (undefined [UNDEF]) and compared the expression of DG, FSP1, and podocyte Wilms' tumor antigen (WT1). RESULTS: WT1 is markedly decreased in NOS versus normal and correlates with the extent of sclerosis. alpha- and beta-DG are maintained in most primary and secondary FSGS cases. In contrast, alpha-DG is significantly decreased in UNDEF, supporting a diagnosis of MCD. Furthermore, follow-up shows remission or decreased proteinuria in four of six of these UNDEF cases in response to therapy. Interstitial FSP1 is numerically highest in COLL but is only rarely found in tubules or podocytes in any other forms of FSGS. CONCLUSIONS: We conclude that increased FSP1 may be a marker of the aggressive course of collapsing FSGS. Furthermore, DG staining is a useful adjunct to assist in distinction of FSGS versus MCD in biopsies without defining lesions. PMID- 19808231 TI - Key comorbid conditions that are predictive of survival among hemodialysis patients. AB - BACKGROUND AND OBJECTIVES: Abstracting information about comorbid illnesses from the medical record can be time-consuming, particularly when a large number of conditions are under consideration. We sought to determine which conditions are most prognostic and whether comorbidity continues to contribute to a survival model once laboratory and clinical parameters have been accounted for. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Comorbidity data were abstracted from the medical records of Dialysis Outcomes and Practice Pattern Study (DOPPS) I, II, and III participants using a standardized questionnaire. Models that were composed of different combinations of comorbid conditions and case-mix factors were compared for explained variance (R(2)) and discrimination (c statistic). RESULTS: Seventeen comorbid conditions account for 96% of the total explained variance that would result if 45 comorbidities that were expected to be predictive of survival were added to a demographics-adjusted survival model. These conditions together had more discriminatory power (c statistic 0.67) than age alone (0.63) or serum albumin (0.60) and were equivalent to a combination of routine laboratory and clinical parameters (0.67). The strength of association of the individual comorbidities lessened when laboratory/clinical parameters were added, but all remained significant. The total R(2) of a model adjusted for demographics and laboratory/clinical parameters increased from 0.13 to 0.17 upon addition of comorbidity. CONCLUSIONS: A relatively small list of comorbid conditions provides equivalent discrimination and explained variance for survival as a more extensive characterization of comorbidity. Comorbidity adds to the survival model a modest amount of independent prognostic information that cannot be substituted by clinical/laboratory parameters. PMID- 19808232 TI - Restricting ciprofloxacin-induced resistant variant formation in biofilm of Klebsiella pneumoniae B5055 by complementary bacteriophage treatment. AB - OBJECTIVES: The in vitro efficacy of lytic bacteriophage as a therapeutic agent against Klebsiella pneumoniae biofilm, alone or in combination with ciprofloxacin, was studied. The pathogenic potential of the resistant variants formed during the treatment was evaluated. METHODS: A lytic bacteriophage in combination with ciprofloxacin was used for the treatment of K. pneumoniae B5055 biofilm. The efficacy and the frequency of resistant variant formation were estimated after respective treatments. The resistant variants were characterized for their virulence potential. RESULTS: Bacteriophage alone was able to eradicate the biofilm effectively and no significant difference was observed in its ability to eradicate biofilm in combination with ciprofloxacin. However, combination treatment using ciprofloxacin and bacteriophage significantly arrested the emergence of resistant variants. The small number of variants that developed had a lower propensity to form biofilms, produced small amounts of cell-associated capsular polysaccharide and demonstrated increased susceptibility to mouse peritoneal macrophages. Altered morphology and changed pattern of the outer membrane proteins of bacterial isolates were also observed. CONCLUSIONS: The combination treatment not only killed the bacteria, but also restricted the formation of resistant variants significantly as compared with individual treatments. Hence, a combination of bacteriophage and ciprofloxacin offers an effective strategy to combat the emergence of treatment-associated resistance. PMID- 19808233 TI - Novel genetic environment of plasmid-mediated quinolone resistance gene qnrB2 in Salmonella Bredeney from poultry. PMID- 19808234 TI - Plasmid-mediated ArmA and RmtB 16S rRNA methylases in Escherichia coli isolated from chickens. PMID- 19808235 TI - Diversity of antimicrobial resistance pheno- and genotypes of methicillin resistant Staphylococcus aureus ST398 from diseased swine. AB - OBJECTIVES: Fifty-four methicillin-resistant Staphylococcus aureus (MRSA) ST398 isolates from unrelated diseased swine collected all over Germany were comparatively investigated for their antimicrobial resistance and virulence properties, and for their genomic relatedness. METHODS: MICs of 30 antimicrobial agents were determined by broth microdilution. Resistance and virulence genes were detected via a diagnostic DNA microarray and specific PCRs. The genomic relationships were determined by ApaI-PFGE, spa typing and SCCmec typing. RESULTS: Twenty-two distinct resistance patterns were observed. All 54 isolates were tetracycline resistant, mediated by tet(M), tet(K) and/or tet(L), with 14 isolates being only resistant to beta-lactam antibiotics and tetracyclines. Trimethoprim resistance, seen in 28 isolates, was mostly due to the gene dfrK or dfrG. Among the 24 macrolide/lincosamide-resistant isolates, the genes erm(A), erm(B) and/or erm(C) were detected. The two chloramphenicol/florfenicol-resistant isolates harboured the gene fexA. The eight gentamicin-resistant isolates carried the gene aacA/aphD. Fifty-three isolates harboured SCCmec type V elements while the remaining one carried mecA and ugpQ, but no recombinase genes. All isolates were PVL negative, but one and three isolates, respectively, were positive for the enterotoxin B and enterotoxin K and Q genes. Eight different spa types were identified with t011 being the most predominant. Six ApaI-PFGE clusters with up to nine individual patterns were detected. CONCLUSIONS: MRSA ST398 isolates varied slightly in their virulence properties and spa types but differed distinctly in their antimicrobial resistance pheno- and genotypes as well as their ApaI-PFGE patterns. These data underline the ability of ST398 to acquire genetic material that might increase antimicrobial resistance and virulence. PMID- 19808236 TI - Epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) among residents of nursing homes in Belgium. AB - OBJECTIVES: A national survey was conducted to determine the prevalence, risk factors and molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) carriage among nursing home (NH) residents in Belgium. METHODS: A random stratified, cross-sectional prevalence survey was conducted in NH residents who were screened for MRSA carriage by multisite enriched culture. Characteristics of NHs and residents were collected by a questionnaire survey and analysed by two stage logistic regression modelling. MRSA isolates were genotyped by PFGE, staphylococcal cassette chromosome mec (SCCmec) typing, multilocus sequence typing (MLST) and resistance genes. RESULTS: Of 2953 residents screened in 60 NHs, 587 (19.9%) were MRSA carriers. Risk factors included hospital contact, antibiotic exposure, impaired mobility and skin lesions at the resident level, and lack of MRSA surveillance, lack of antibiotic therapeutic formulary and the combination of less-developed infection control activities and a high ratio of physicians to residents at the institution level. MRSA isolates showed eight major types, three of which were predominant: B2-ST45-SCCmec IV (49%; where ST stands for sequence type); A21-ST8-SCCmec IV (13%); and A20-ST8-SCCmec IV (10%). Each was recovered in 55, 21 and 25 NHs, respectively. The geographical distribution of NH genotypes paralleled that of acute-care hospitals. CONCLUSIONS: A high prevalence of MRSA carriage in NH residents was associated with hospital care, co-morbidities and less-developed coordination of institutional care. The predominant MRSA strains from NH residents and hospitalized patients of the same area were identical. Strengthening and coordination of MRSA surveillance and control activities are warranted within and between NHs and hospitals. PMID- 19808237 TI - Prevalence of antimicrobial-resistant bacteria isolated from older versus younger hospitalized adults: results of a two-centre study. AB - OBJECTIVES: To compare the proportion of antimicrobial-resistant strains among bacterial isolates from younger and older hospital patients and to quantify changes in the proportion of antimicrobial-resistant strains in both groups over time. PATIENTS AND METHODS: A retrospective analysis of microbiology data from two centres in Maryland and Chicago was performed. Adult hospital inpatients with positive clinical cultures for specific antimicrobial-resistant bacterial pathogens between 1999 and 2005 (55 427 isolates) were included. The proportions of isolates not susceptible to specific antimicrobial agents were compared between patients > or =65 and <65 years. Additional analyses examined temporal trends in the frequency of resistance and the frequency of resistance among the oldest patients (> or =80 years), in bacteria isolated from blood cultures and in bacteria obtained from intensive care unit patients. RESULTS: Heterogeneity was observed in the frequency of resistance among different bacteria between older and younger patients, between the two centres and over the study period. Staphylococcus aureus isolates were more likely to be resistant to methicillin when obtained from older patients at Chicago (50.9% versus 40.9%; P < 0.001). In contrast, younger patients yielded a greater proportion of enterococci resistant to vancomycin at Maryland (19.4% versus 16.5%; P = 0.009). Results were variable when resistance to fluoroquinolones, cephalosporins and imipenem were compared for Pseudomonas aeruginosa, Escherichia coli and Klebsiella spp. CONCLUSIONS: Overall, advanced patient age was not uniformly associated with a greater likelihood of antimicrobial resistance among all bacterial pathogens. Moreover, the frequency of resistance in older and younger patients varied considerably at the two sites over the study period. Variability in the frequency of resistance precludes simplistic conclusions regarding the relationship between age and resistance. PMID- 19808238 TI - Fuller albright: the consummate clinical investigator. PMID- 19808239 TI - Long-term outcome of children with steroid-sensitive idiopathic nephrotic syndrome. PMID- 19808240 TI - The need for a children's oncology group-oriented approach to advance the care of children with Idiopathic nephrotic syndrome. PMID- 19808241 TI - Weight loss interventions in chronic kidney disease: a systematic review and meta analysis. AB - BACKGROUND AND OBJECTIVES: Obesity is an independent risk factor for development and progression of chronic kidney disease (CKD). We conducted a systematic review to assess the benefits of intentional weight loss in patients with non-dialysis dependent CKD and glomerular hyperfiltration. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We searched MEDLINE, SCOPUS, and conference proceedings for randomized, controlled trials and observational studies that examined various surgical and nonsurgical interventions (diet, exercise, and/or antiobesity agents) in adult patients with CKD. Results were summarized using random-effects model. RESULTS: Thirteen studies were included. In patients with CKD, body mass index (BMI) decreased significantly (weighted mean difference [WMD] -3.67 kg/m(2); 95% confidence interval [CI] -6.56 to -0.78) at the end of the study period with nonsurgical interventions. This was associated with a significant decrease in proteinuria (WMD -1.31 g/24 h; 95% CI -2.11 to -0.51) and systolic BP with no further decrease in GFR during a mean follow-up of 7.4 mo. In morbidly obese individuals (BMI >40 kg/m(2)) with glomerular hyperfiltration (GFR >125 ml/min), surgical interventions decreased BMI, which resulted in a decrease in GFR (WMD -25.56 ml/min; 95% CI -36.23 to -14.89), albuminuria, and systolic BP. CONCLUSIONS: In smaller, short-duration studies in patients with CKD, nonsurgical weight loss interventions reduce proteinuria and BP and seem to prevent further decline in renal function. In morbidly obese individuals with glomerular hyperfiltration, surgical interventions normalize GFR and reduce BP and microalbuminuria. Larger, long-term studies to analyze renal outcomes such as development of ESRD are needed. PMID- 19808242 TI - Scaling of measured glomerular filtration rate in kidney donor candidates by anthropometric estimates of body surface area, body water, metabolic rate, or liver size. AB - BACKGROUND AND OBJECTIVES: GFR is scaled to body surface area (S), whereas hemodialysis dosage is scaled to total body water (V). Scaling to metabolic rate (M) or liver size (L) has also been proposed. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: In 1551 potential kidney donors (662 men and 889 women) for whom GFR had been estimated from (125)I-iothalamate clearance (iGFR) between the years 1973 and 2005, iGFR scaling was examined. Scaling was to estimates of S, V, M, or L. The study looked at the variation of iGFR by gender, age, S, V, M, and L within the study population. RESULTS: In multiple regression analysis, neither gender nor race was significantly associated with iGFR after controlling for height, weight, and age. Raw iGFR averaged 122 +/- 23 ml/min in men and 106 +/- 21 ml/min in women (P < 0.001). In an adjusted analysis, iGFR scaled to S or L was similar for men and women (NS), whereas iGFR scaled to either V or M was substantially different between the genders (P < 0.001). When the patients by gender were divided into five quintiles of V or S, the iGFR-V ratio varied more with body size than iGFR scaled to the other measures. CONCLUSIONS: iGFR scaled to S or L was similar in men and women. Scaling to either M or V resulted in a sizeable gender difference, whereas scaling to V led to markedly different values of iGFR across body size. PMID- 19808243 TI - Long-term outcome of biopsy-proven, frequently relapsing minimal-change nephrotic syndrome in children. AB - BACKGROUND AND OBJECTIVES: Frequently relapsing and steroid-dependent minimal change nephrotic syndrome (MCNS) that originates in childhood can persist after puberty in >20% of patients. These patients require immunosuppressive treatment during several decades of their life. We examined long-term adverse effects of persistent nephrotic syndrome and immunosuppressive medications, focusing on renal function, growth, obesity, osteoporosis, hypertension, ocular complications, and fertility in adult patients with biopsy-proven childhood-onset MCNS. Molecular analysis was performed to evaluate a possible association of a complicated course of MCNS with podocyte gene mutations. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We performed a prospective clinical examination of 15 adult patients that included serum and urine analysis; dual-energy x-ray absorptiometry; ophthalmologic examination; semen examination; and molecular analysis of NPHS1, NPHS2, CD2AP, and ACTN4 genes. RESULTS: All patients had normal GFR. Most frequent long-term complications were hypertension (in seven of 15 patients) and osteoporosis in one third of patients. Oligozoospermia was found in one patient, reduced sperm motility in four of eight patients, and teratozoospermia in six of eight patients. Ophthalmologic examination revealed myopia in 10 of 15 patients and cataract in three of 15 patients. CONCLUSIONS: Children with MCNS that persists after puberty are at risk for complications such as osteoporosis, hypertension, cataract, and sperm abnormalities. Our study underscores a need for more effective and less toxic therapies for relapsing MCNS. PMID- 19808244 TI - Is maximum conservative management an equivalent treatment option to dialysis for elderly patients with significant comorbid disease? AB - BACKGROUND AND OBJECTIVES: There is ongoing growth of elderly populations with ESRD in Western Europe and North America. In our center, we offer an alternative care pathway of 'maximum conservative management' (MCM) to patients who elect not to start dialysis, often because of a heavy burden of comorbid illness and advanced age. The objective of our study was to compare clinical outcomes for patients who had ESRD and chose either MCM or renal replacement therapy (RRT). DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This is an observational study of a single-center cohort in the United Kingdom that evaluating 202 elderly (> or =70 yr) patients who had ESRD and had chosen either MCM (n = 29) or RRT (n = 173). We report survival, hospitalization rates, and location of death for this cohort. Survival was measured from a standardized 'threshold' estimated GFR of 10.8 ml/min per 1.73 m(2). RESULTS: Median survival, including the first 90 d, was 37.8 mo (range 0 to 106 mo) for RRT patients and 13.9 mo (range 2 to 44) for MCM patients (P < 0.01). RRT patients had higher rates of hospitalization (0.069 [95% confidence interval (CI) 0.068 to 0.070]) versus 0.043 [95% CI 0.040 to 0.047] hospital days/patient-days survived) compared with MCM patients. MCM patients were significantly more likely to die at home or in a hospice (odds ratio 4.15; 95% CI 1.67 to 10.25). A survey of the literature describing elderly ESRD outcomes is also presented. CONCLUSIONS: Dialysis prolongs survival for elderly patients who have ESRD with significant comorbidity by approximately 2 yr; however, patients who choose MCM can survive a substantial length of time, achieving similar numbers of hospital-free days to patients who choose hemodialysis. PMID- 19808245 TI - Mineral metabolism and inflammation in chronic kidney disease patients: a cross sectional study. AB - BACKGROUND AND OBJECTIVES: Mineral metabolism abnormalities and inflammation are concerns in chronic kidney disease (CKD). Interrelationships among these parameters have not been analyzed. DESIGN, SETTING, PARTICIPANTS, AND MEASUREMENTS: The study included 133 patients with CKD not on dialysis and not receiving calcium (Ca) supplements, phosphate binders, or vitamin D. Estimated GFR (eGFR) was 34.1 +/- 6.8 ml/min/1.73 m(2); 107 participants had stage 3 CKD, and 26 had stage 4. RESULTS: Patients were classified by tertiles of Ca, phosphorus (P), Ca-P product (Ca x P), and parathyroid hormone (PTH). After adjustment for age, gender, and eGFR, the levels of C-reactive protein (CRP) and IL-6 (IL-6) of the third tertile of P, Ca x P, and PTH were significantly higher than those of the first and second tertiles. Serum P and Ca x P directly correlated with CRP and IL-6, whereas HDL-cholesterol and eGFR inversely correlated with the levels of the inflammatory parameters. After partial correlation analysis, the previous associations between CRP and eGFR, and serum P, as well as the relationship between IL-6 and eGFR, and serum P, remained significant. Multiple regression analysis demonstrated that eGFR and serum P were independently associated with CRP and IL-6. Finally, logistic regression analysis using the presence/absence of an inflammatory state as the dependent variable showed that eGFR was a protective factor, whereas serum P was an independent risk factor for the presence of an inflammatory state. CONCLUSIONS: Elevated serum P might play a role in the development of inflammation in CKD. PMID- 19808246 TI - Can we personalize treatment for kidney diseases? AB - The idea of individualizing therapies to obtain optimal clinical results is not new but has only recently been applied to kidney diseases. Nonetheless, kidney disorders present a variety of opportunities to personalize medicine. Here, the heterogeneity of kidney disorders is reviewed to provide a rationale for pursuing personalized medicine. Data on adjusting therapy on the basis of pharmacogenetics/genomics and pharmacodynamics are summarized to demonstrate where the field is, and biomarker studies that reflect the future of personalized medicine are discussed. The goal of this review is to demonstrate that we can personalize therapy for kidney diseases but that considerable investment in new research will be required for personalized medicine to be routinely used in nephrology clinics. PMID- 19808249 TI - Native and aspirin-triggered lipoxins control innate immunity by inducing proteasomal degradation of TRAF6. PMID- 19808248 TI - Ir-CPI, a coagulation contact phase inhibitor from the tick Ixodes ricinus, inhibits thrombus formation without impairing hemostasis. AB - Blood coagulation starts immediately after damage to the vascular endothelium. This system is essential for minimizing blood loss from an injured blood vessel but also contributes to vascular thrombosis. Although it has long been thought that the intrinsic coagulation pathway is not important for clotting in vivo, recent data obtained with genetically altered mice indicate that contact phase proteins seem to be essential for thrombus formation. We show that recombinant Ixodes ricinus contact phase inhibitor (Ir-CPI), a Kunitz-type protein expressed by the salivary glands of the tick Ixodes ricinus, specifically interacts with activated human contact phase factors (FXIIa, FXIa, and kallikrein) and prolongs the activated partial thromboplastin time (aPTT) in vitro. The effects of Ir-CPI were also examined in vivo using both venous and arterial thrombosis models. Intravenous administration of Ir-CPI in rats and mice caused a dose-dependent reduction in venous thrombus formation and revealed a defect in the formation of arterial occlusive thrombi. Moreover, mice injected with Ir-CPI are protected against collagen- and epinephrine-induced thromboembolism. Remarkably, the effective antithrombotic dose of Ir-CPI did not promote bleeding or impair blood coagulation parameters. To conclude, our results show that a contact phase inhibitor is an effective and safe antithrombotic agent in vivo. PMID- 19808250 TI - Mast cell-derived particles deliver peripheral signals to remote lymph nodes. AB - During infection, signals from the periphery are known to reach draining lymph nodes (DLNs), but how these molecules, such as inflammatory cytokines, traverse the significant distances involved without dilution or degradation remains unclear. We show that peripheral mast cells, upon activation, release stable submicrometer heparin-based particles containing tumor necrosis factor and other proteins. These complexes enter lymphatic vessels and rapidly traffic to the DLNs. This physiological drug delivery system facilitates communication between peripheral sites of inflammation and remote secondary lymphoid tissues. PMID- 19808251 TI - CD1-restricted adaptive immune responses to Mycobacteria in human group 1 CD1 transgenic mice. AB - Group 1 CD1 (CD1a, CD1b, and CD1c)-restricted T cells recognize mycobacterial lipid antigens and are found at higher frequencies in Mycobacterium tuberculosis (Mtb)-infected individuals. However, their role and dynamics during infection remain unknown because of the lack of a suitable small animal model. We have generated human group 1 CD1 transgenic (hCD1Tg) mice that express all three human group 1 CD1 isoforms and support the development of group 1 CD1-restricted T cells with diverse T cell receptor usage. Both mycobacterial infection and immunization with Mtb lipids elicit group 1 CD1-restricted Mtb lipid-specific T cell responses in hCD1Tg mice. In contrast to CD1d-restricted NKT cells, which rapidly respond to initial stimulation but exhibit anergy upon reexposure, group 1 CD1-restricted T cells exhibit delayed primary responses and more rapid secondary responses, similar to conventional T cells. Collectively, our data demonstrate that group 1 CD1-restricted T cells participate in adaptive immune responses upon mycobacterial infection and could serve as targets for the development of novel Mtb vaccines. PMID- 19808255 TI - Dendritic cells are crucial for maintenance of tertiary lymphoid structures in the lung of influenza virus-infected mice. AB - Tertiary lymphoid organs (TLOs) are organized aggregates of B and T cells formed in postembryonic life in response to chronic immune responses to infectious agents or self-antigens. Although CD11c+ dendritic cells (DCs) are consistently found in regions of TLO, their contribution to TLO organization has not been studied in detail. We found that CD11c(hi) DCs are essential for the maintenance of inducible bronchus-associated lymphoid tissue (iBALT), a form of TLO induced in the lungs after influenza virus infection. Elimination of DCs after the virus had been cleared from the lung resulted in iBALT disintegration and reduction in germinal center (GC) reactions, which led to significantly reduced numbers of class-switched plasma cells in the lung and bone marrow and reduction in protective antiviral serum immunoglobulins. Mechanistically, DCs isolated from the lungs of mice with iBALT no longer presented viral antigens to T cells but were a source of lymphotoxin (LT) beta and homeostatic chemokines (CXCL-12 and 13 and CCL-19 and -21) known to contribute to TLO organization. Like depletion of DCs, blockade of LTbeta receptor signaling after virus clearance led to disintegration of iBALT and GC reactions. Together, our data reveal a previously unappreciated function of lung DCs in iBALT homeostasis and humoral immunity to influenza virus. PMID- 19808254 TI - Transforming growth factor beta is dispensable for the molecular orchestration of Th17 cell differentiation. AB - Interleukin (IL)-17-producing T helper (Th17) cells play a critical role in the pathophysiology of several autoimmune disorders. The differentiation of Th17 cells requires the simultaneous presence of an unusual combination of cytokines: IL-6, a proinflammatory cytokine, and transforming growth factor (TGF) beta, an antiinflammatory cytokine. However, the molecular mechanisms by which TGF-beta exerts its effects on Th17 cell differentiation remain elusive. We report that TGF-beta does not directly promote Th17 cell differentiation but instead acts indirectly by blocking expression of the transcription factors signal transducer and activator of transcription (STAT) 4 and GATA-3, thus preventing Th1 and Th2 cell differentiation. In contrast, TGF-beta had no effect on the expression of retinoic acid receptor-related orphan nuclear receptor gammat, a Th17-specific transcription factor. Interestingly, in Stat-6(-/-)T-bet(-/-) mice, which are unable to generate Th1 and Th2 cells, IL-6 alone was sufficient to induce robust differentiation of Th17 cells, whereas TGF-beta had no effect, suggesting that TGF-beta is dispensable for Th17 cell development. Consequently, BALB/c Stat-6(-/ )T-bet(-/-) mice, but not wild-type BALB/c mice, were highly susceptible to the development of experimental autoimmune encephalomyelitis, which could be blocked by anti-IL-17 antibodies but not by anti-TGF-beta antibodies. Collectively, these data provide evidence that TGF-beta is not directly required for the molecular orchestration of Th17 cell differentiation. PMID- 19808256 TI - Staphylococcus aureus synthesizes adenosine to escape host immune responses. AB - Staphylococcus aureus infects hospitalized or healthy individuals and represents the most frequent cause of bacteremia, treatment of which is complicated by the emergence of methicillin-resistant S. aureus. We examined the ability of S. aureus to escape phagocytic clearance in blood and identified adenosine synthase A (AdsA), a cell wall-anchored enzyme that converts adenosine monophosphate to adenosine, as a critical virulence factor. Staphylococcal synthesis of adenosine in blood, escape from phagocytic clearance, and subsequent formation of organ abscesses were all dependent on adsA and could be rescued by an exogenous supply of adenosine. An AdsA homologue was identified in the anthrax pathogen, and adenosine synthesis also enabled escape of Bacillus anthracis from phagocytic clearance. Collectively, these results suggest that staphylococci and other bacterial pathogens exploit the immunomodulatory attributes of adenosine to escape host immune responses. PMID- 19808257 TI - Loss of matrix metalloproteinase 2 in platelets reduces arterial thrombosis in vivo. AB - Platelet activation at a site of vascular injury is essential for the arrest of bleeding; however, excessive platelet activation at a site of arterial damage can result in the unwarranted formation of arterial thrombi, precipitating acute myocardial infarction, or ischemic stroke. Activation of platelets beyond the purpose of hemostasis may occur when substances facilitating thrombus growth and stability accumulate. Human platelets contain matrix metalloproteinase 2 (MMP-2) and release it upon activation. Active MMP-2 amplifies the platelet aggregation response to several agonists by potentiating phosphatidylinositol 3-kinase activation. Using several in vivo thrombosis models, we show that the inactivation of the MMP-2 gene prevented thrombosis induced by weak, but not strong, stimuli in mice but produced only a moderate prolongation of the bleeding time. Moreover, using cross-transfusion experiments and wild-type/MMP-2(-/-) chimeric mice, we show that it is platelet-derived MMP-2 that facilitates thrombus formation. Finally, we show that platelets activated by a mild vascular damage induce thrombus formation at a downstream arterial injury site by releasing MMP-2. Thus, platelet-derived MMP-2 plays a crucial role in thrombus formation by amplifying the response of platelets to weak activating stimuli. These findings open new possibilities for the prevention of thrombosis by the development of MMP-2 inhibitors. PMID- 19808258 TI - Phosphoinositide-dependent kinase 1 controls migration and malignant transformation but not cell growth and proliferation in PTEN-null lymphocytes. AB - In normal T cell progenitors, phosphoinositide-dependent kinase l (PDK1)-mediated phosphorylation and activation of protein kinase B (PKB) is essential for the phosphorylation and inactivation of Foxo family transcription factors, and also controls T cell growth and proliferation. The current study has characterized the role of PDK1 in the pathology caused by deletion of the tumor suppressor phosphatase and tensin homologue deleted on chromosome 10 (PTEN). PDK1 is shown to be essential for lymphomagenesis caused by deletion of PTEN in T cell progenitors. However, PTEN deletion bypasses the normal PDK1-controlled signaling pathways that determine thymocyte growth and proliferation. PDK1 does have important functions in PTEN-null thymocytes, notably to control the PKB-Foxo signaling axis and to direct the repertoire of adhesion and chemokine receptors expressed by PTEN-null T cells. The results thus provide two novel insights concerning pathological signaling caused by PTEN loss in lymphocytes. First, PTEN deletion bypasses the normal PDK1-controlled metabolic checkpoints that determine cell growth and proliferation. Second, PDK1 determines the cohort of chemokine and adhesion receptors expressed by PTEN-null cells, thereby controlling their migratory capacity. PMID- 19808261 TI - Mickie Bhatia: Embryonic stem cells come of age. PMID- 19808260 TI - Subtilase cytotoxin cleaves newly synthesized BiP and blocks antibody secretion in B lymphocytes. AB - Shiga-toxigenic Escherichia coli (STEC) use subtilase cytotoxin (SubAB) to interfere with adaptive immunity. Its inhibition of immunoglobulin secretion is both rapid and profound. SubAB favors cleavage of the newly synthesized immunoglobulin heavy chain-binding protein (BiP) to yield a C-terminal fragment that contains BiP's substrate-binding domain. In the absence of its regulatory nucleotide-binding domain, the SubAB-cleaved C-terminal BiP fragment remains tightly bound to newly synthesized immunoglobulin light chains, resulting in retention of light chains in the endoplasmic reticulum (ER). Immunoglobulins are thus detained in the ER, making impossible the secretion of antibodies by SubAB treated B cells. The inhibitory effect of SubAB is highly specific for antibody secretion, because other secretory proteins such as IL-6 are released normally from SubAB-treated B cells. Although SubAB also causes BiP cleavage in HepG2 hepatoma cells, (glyco)protein secretion continues unabated in SubAB-exposed HepG2 cells. This specific block in antibody secretion is a novel means of immune evasion for STEC. The differential cleavage of newly synthesized versus "aged" BiP by SubAB in the ER provides insight into the architecture of the ER compartments involved. PMID- 19808262 TI - The inaugural issue of Circulation: Heart Failure. PMID- 19808259 TI - T-bet-dependent S1P5 expression in NK cells promotes egress from lymph nodes and bone marrow. AB - During a screen for ethylnitrosourea-induced mutations in mice affecting blood natural killer (NK) cells, we identified a strain, designated Duane, in which NK cells were reduced in blood and spleen but increased in lymph nodes (LNs) and bone marrow (BM). The accumulation of NK cells in LNs reflected a decreased ability to exit into lymph. This strain carries a point mutation within Tbx21 (T bet), which generates a defective protein. Duane NK cells have a 30-fold deficiency in sphingosine-1-phosphate receptor 5 (S1P5) transcript levels, and S1P5-deficient mice exhibit an egress defect similar to Duane. Chromatin immunoprecipitation confirms binding of T-bet to the S1pr5 locus. S1P-deficient mice exhibit a more severe NK cell egress block, and the FTY720-sensitive S1P1 also plays a role in NK cell egress from LNs. S1P5 is not inhibited by CD69, a property that may facilitate trafficking of activated NK cells to effector sites. Finally, the accumulation of NK cells within BM of S1P-deficient mice was associated with reduced numbers in BM sinusoids, suggesting a role for S1P in BM egress. In summary, these findings identify S1P5 as a T-bet-induced gene that is required for NK cell egress from LNs and BM. PMID- 19808263 TI - Blood urea nitrogen and serum creatinine: not married in heart failure. PMID- 19808264 TI - The tumultuous journey of nesiritide: past, present, and future. PMID- 19808266 TI - Effects of the oral direct renin inhibitor aliskiren in patients with symptomatic heart failure. AB - BACKGROUND: Loss of negative feedback inhibition of renin release during chronic treatment with an angiotensin-converting enzyme (ACE) inhibitor leads to a compensatory rise in renin secretion and downstream components of the renin angiotensin-aldosterone (RAAS) cascade. This may overcome ACE inhibition but should be blocked by a direct renin inhibitor. We studied the effects of adding the direct renin inhibitor aliskiren to an ACE inhibitor in patients with heart failure. METHODS AND RESULTS: Patients with New York Heart Association class II to IV heart failure, current or past history of hypertension, and plasma brain natriuretic peptide (BNP) concentration >100 pg/mL who had been treated with an ACE inhibitor (or angiotensin receptor blocker) and beta-blocker were randomized to 3 months of treatment with placebo (n=146) or aliskiren 150 mg/d (n=156). The primary efficacy outcome was the between-treatment difference in N-terminal pro BNP (NT-proBNP). Patients' mean age was 68 years, mean ejection fraction was 31%, and mean+/-SD systolic blood pressure was 129+/-17.4 mm Hg. Sixty-two percent of the patients were in New York Heart Association functional class II, and 33% were taking an aldosterone antagonist. Plasma NT-proBNP rose by 762+/-6123 pg/mL with placebo and fell by 244+/-2025 pg/mL with aliskiren (P=0.0106). BNP and urinary (but not plasma) aldosterone were also reduced by aliskiren. Clinically important differences in blood pressure and biochemistry were not seen between aliskiren and placebo. CONCLUSIONS: Addition of aliskiren to an ACE inhibitor (or angiotensin receptor blocker) and beta-blocker had favorable neurohumoral effects in heart failure and appeared to be well tolerated. PMID- 19808267 TI - Admission or changes in renal function during hospitalization for worsening heart failure predict postdischarge survival: results from the Outcomes of a Prospective Trial of Intravenous Milrinone for Exacerbations of Chronic Heart Failure (OPTIME-CHF). AB - BACKGROUND: Admission measures of renal function (blood urea nitrogen [BUN], estimated glomerular filtration rate [eGFR]) in patients hospitalized for worsening heart failure are predictors of in-hospital outcomes. Less is known about the changes and relationships among these variables and the postdischarge survival rate. METHODS AND RESULTS: In a retrospective analysis of 949 patients from the Outcomes of a Prospective Trial of Intravenous Milrinone for Exacerbations of Chronic Heart Failure, we investigated the relation between admission values and changes in BUN and eGFR and rate of death by 60 days after discharge. On admission, median eGFR was 51 mL min(-1) 1.73 m(-2) (interquartile range, 37 to 70 mL min(-1) 1.73 m(-2)), and BUN was 25 mg/dL (interquartile range, 17 to 41 mg/dL). On average, there was a 1.1-mL min(-1) 1.73 m(-2) decrease in eGFR and a 4.7-mg/dL increase in BUN from admission to discharge. By discharge, 12% of patients had a >25% decrease in eGFR, and 39% had a >25% increase in BUN. Although both lower admission eGFR and higher admission BUN were associated with higher risk of death by 60 days after discharge, multivariable adjusted Cox proportional-hazards analysis showed that BUN was a stronger predictor of death by 60 days than was eGFR (chi(2), 11.6 and 0.6 for BUN and eGFR, respectively). Independently of admission values, an increase of >or=10 mg/dL in BUN during hospitalization was associated with worse 60-day survival rate: BUN (per 5-mg/dL increase) had a hazard ratio of 1.08 (95% CI, 1.01 to 1.16). Although milrinone treatment led to a minor improvement in renal function by discharge, the 60-day death and readmission rates were similar between the milrinone and placebo groups. CONCLUSIONS: A substantial number of patients admitted with heart failure have worsening renal function during hospitalization. Higher admission BUN and increasing BUN during hospitalization, independently of admission values, are associated with a worse survival rate. Use of milrinone in these high-risk patients does not improve outcomes despite minor improvements in the renal function. PMID- 19808268 TI - Effect of baseline and changes in systolic blood pressure over time on the effectiveness of valsartan in the Valsartan Heart Failure Trial. AB - BACKGROUND: Low systolic blood pressure (SBP) is a risk factor for adverse outcomes in patients with heart failure (HF). Valsartan improved morbidity rates in the Valsartan Heart Failure Trial (Val-HeFT) despite a reduction in SBP. The aim of the present study was to investigate the relationship between the SBP lowering effects of valsartan and its cardiovascular protective effects in this population. METHODS AND RESULTS: Baseline measurements and changes in SBP at 4 months were related to mortality and morbidity rates. The effects of valsartan on these end points were compared in quartiles of baseline SBP with multivariable Cox proportional hazards regression models that included a test for interaction between the effects of valsartan treatment and baseline SBP and examined the effects of changes in SBP on the valsartan effect. The mean+/-SD baseline SBP in all patients (n=5010) was 124+/-18 mm Hg. Patients in the lowest quartile of SBP (SBP 25% DE in 10% of patients. The presence of DE was related to occurrence of heart failure symptoms (P=0.05) and left ventricular systolic dysfunction (P=0.001). DE was present in all patients with ejection fraction < or =50% but also in 53% (102/192) of patients with preserved ejection fraction (P<0.001); %DE was both inversely related to (r=-0.3; P<0.001) and an independent predictor of ejection fraction (r=-0.4; P<0.001). DE (7%+/-7% of left ventricle) was present in 54 patients who were asymptomatic (and with normal ejection fraction). Over the follow-up period, the annualized adverse cardiovascular event rate in patients with DE exceeded that in patients without DE but did not achieve statistical significance (5.5% versus 3.3%; P=0.5). CONCLUSIONS: In a large HCM cohort, DE was an independent predictor of systolic dysfunction but with only a modest relationship to heart failure symptoms. These data suggest an important role for myocardial fibrosis in the clinical course of HCM patients but are not sufficient at this time to consider DE as an independent risk factor for adverse prognosis. PMID- 19808289 TI - Titin isoforms, extracellular matrix, and global chamber remodeling in experimental dilated cardiomyopathy: functional implications and mechanistic insight. AB - BACKGROUND: Altered titin isoforms may modify cardiac function in heart failure (HF), but the nature of isoform switches and associated functional implications are not well defined. Limited studies have reported an increased compliant isoform (N2BA) expression in human systolic HF. Titin may also modulate stretch regulated responses such as myocardial natriuretic peptide production. METHODS AND RESULTS: We characterized titin isoform expression and extracellular matrix in all 4 cardiac chambers and the left ventricular (LV) epicardium and endocardium in normal dogs (n=6) and those with HF (n=6) due to tachypacing and characterized functional implications at the LV myofiber and chamber level. Recognizing the potential for uncoupling of the extracellular matrix and cardiomyocyte in tachypacing, myocardial natriuretic peptide production, a molecular marker of stretch-regulated responses, was also assessed. All chambers were dilated in HF, but the extracellular matrix was not increased. HF dogs had markedly lower N2BA in the atria and right ventricle. In failing LVs, N2BA was decreased only in the epicardium, where myofiber passive stiffness was increased. However, LV chamber mechanics were driven by the marked LV dilatation, with no increase in LV diastolic stiffness. Natriuretic peptide concentrations increased markedly in the endocardium in relation to increases in LV wall stress. CONCLUSIONS: Tachypacing HF is characterized by decreases in compliant titin isoform expression in the atria, right ventricle, and LV epicardium. However, LV chamber mechanics are principally determined by geometric and extracellular matrix changes rather than titin-based myofiber stiffness in this model. Stretch regulated myocardial responses (natriuretic peptide production) appeared intact, suggesting that the mechanotransduction role of titin was not impaired in HF. PMID- 19808290 TI - Mechanical circulatory support: registering a therapy in evolution. PMID- 19808291 TI - The role of digoxin in the treatment of heart failure. PMID- 19808292 TI - The contemporary use of digoxin for the treatment of heart failure. PMID- 19808293 TI - Phosphodiesterase type 5 inhibition: a support of the left ventricular assist device bridge to transplant. PMID- 19808294 TI - PDE5A inhibitor treatment of persistent pulmonary hypertension after mechanical circulatory support. AB - BACKGROUND: Pulmonary hypertension (PH) secondary to left heart failure portends a poor prognosis and is a relative contraindication to heart transplantation at many centers. We tested the hypothesis that when PH persists after adequate left ventricle unloading via recent left ventricular assist device (LVAD) therapy, phosphodiesterase type 5A inhibition would decrease PH in this population. METHODS AND RESULTS: We performed an open-label clinical trial using control patients not receiving therapy. Between 1999 and 2007, 138 consecutive patients undergoing cardiac transplantation evaluation with advanced left ventricular dysfunction, an elevated pulmonary capillary wedge pressure, and PH (defined by a pulmonary vascular resistance (PVR) >3 Woods Units), were treated with LVAD therapy. Fifty-eight of these patients reduced their pulmonary capillary wedge pressure to a value <15 mm Hg (11.8+/-2.0 mm Hg from baseline 23.2+/-6.2 mm Hg) 1 to 2 weeks after LVAD implantation, but despite this improvement, the PVR of these patients was only minimally affected (5.65+/-3.00 to 5.39+/-1.78 Wood Units). Twenty-six consecutive patients from this group with persistently elevated PVR were started on oral phosphodiesterase type 5A inhibition with sildenafil and titrated to an average of dose of 51.9 mg by mouth 3 times per day. The average PVR in the sildenafil-treated group fell from 5.87+/-1.93 to 2.96+/-0.92 Wood Units (P<0.001) and the mean pulmonary artery pressure fell from 36.5+/-8.6 to 24.3+/-3.6 mm Hg (P<0.0001) and was significantly lower when compared with the 32 LVAD recipients not receiving sildenafil at weeks 12 to 15 after the initial post-LVAD hemodynamic measurements (13 to 17 weeks post-LVAD implantation). In addition, hemodynamic measurements of right ventricular function in sildenafil-treated patients was also improved compared with patients not receiving sildenafil. CONCLUSIONS: In patients with persistent PH after recent LVAD placement, phosphodiesterase type 5A inhibition in this open-label trial resulted in a significant decrease in PVR when compared with control patients. PMID- 19808295 TI - Absence of exercise capacity improvement after exercise training program: a strong prognostic factor in patients with chronic heart failure. AB - BACKGROUND: Exercise training is established as adjuvant therapy for chronic heart failure, but the prognostic value of improvement in exercise capacity after exercise training has never been evaluated. METHODS AND RESULTS: In this prospective bicentric study, all chronic heart failure patients with left ventricular ejection fraction <45% who underwent an exercise training program in a cardiac rehabilitation center between January 2004 and September 2006 were consecutively included. Improvement in exercise capacity was assessed by change in peak oxygen consumption (deltaPVo(2)) and in PVo(2) expressed as a percentage of predicted PVo(2) (delta%PPVo(2)) measured before and after the training program. We included 155 patients (54+/-12 years old, male 81%, left ventricular ejection fraction=29.5+/-7.1%). Patients underwent 20 (10-30) training sessions. PVo(2) and %PPVo(2) were significantly increased after the training program (14% and 13%, respectively, P<0.001 for both). After 16+/-6 months follow-up, 27 patients had a cardiac event (death [n=12], cardiac transplantation [n=5], hospitalization for acute heart failure [n=10]). Univariate analysis revealed that among 17 significant predictors of cardiac events, the 2 more powerful ones were level of B-type natriuretic peptide at baseline (P<0.0001) and improvement in exercise capacity as assessed by deltaPVo(2) and delta%PPVo(2) (P<0.0001). Multivariate analysis revealed B-type natriuretic peptide level and delta%PPVo(2) as only independent predictive factors of outcome (P=0.01). The risk ratio of cardiac events for nonresponse versus response to the training program (defined as median delta%PPVo(2)<6%) was 8.2 (P=0.0006). CONCLUSIONS: Among patients with chronic heart failure, the lack of improvement in exercise capacity after an exercise training program has strong prognostic value for adverse cardiac events independent of classical predictive factors such as left ventricular ejection fraction, New York Heart Association class, and B-type natriuretic peptide level. PMID- 19808296 TI - Determinants of ventilatory efficiency in heart failure: the role of right ventricular performance and pulmonary vascular tone. AB - BACKGROUND: Ventilatory efficiency, right ventricular (RV) function, and secondary pulmonary hypertension are each prognostic indicators in patients with heart failure due to left ventricular systolic dysfunction, but the relationships among these variables have not been comprehensively investigated. In this study, we hypothesized that inefficient ventilation during exercise, as defined by an abnormally steep relationship between ventilation and carbon dioxide output (Ve/Vco(2) slope), may be a marker of secondary pulmonary hypertension and RV dysfunction in heart failure. METHODS AND RESULTS: A cohort of patients with systolic heart failure (mean+/-SD age, 58+/-13 years; left ventricular ejection fraction, 0.27+/-0.05; peak oxygen uptake, 11.2+/-3.2 mL kg(-1) min(-1)) underwent incremental cardiopulmonary exercise testing with simultaneous hemodynamic monitoring and first-pass radionuclide ventriculography before and after 12 weeks of treatment with sildenafil, a selective pulmonary vasodilator, or placebo. Ve/Vco(2) slope was positively related to rest and exercise pulmonary vascular resistance (R=0.39 and R=0.60, respectively) and rest pulmonary capillary wedge pressure (R=0.49, P<0.005 for all) and weakly indirectly related to peak exercise RV ejection fraction (R=-0.29, P=0.03). Over the 12-week study period, Ve/Vco(2) slope fell 8+/-3% (P=0.02) with sildenafil and was unchanged with placebo. Changes in Ve/Vco(2) slope correlated with changes in exercise pulmonary vascular resistance (R=0.69, P<0.001) and rest and exercise RV ejection fraction (R=-0.58 and -0.40, respectively, both P<0.05). CONCLUSIONS: In patients with systolic heart failure and secondary pulmonary hypertension, ventilatory efficiency is closely related to RV function and pulmonary vascular tone during exercise. PMID- 19808297 TI - Discordant short- and long-term outcomes associated with diabetes in patients with heart failure: importance of age and sex: a population study of 5.1 million people in Scotland. AB - BACKGROUND: Diabetes and heart failure frequently coexist. Our aim was to assess the association between diabetes and short- and long-term outcomes in all patients admitted to the hospital for the first time with heart failure in Scotland between 1986 and 2003. METHODS AND RESULTS: A total of 116 556 patients were studied, of whom 13% (n=15 161) had a diagnosis of diabetes. At 30 days, diabetes was associated with a lower case fatality. By 1 year, the association between diabetes and better outcome was reversed, and diabetes was a significant independent predictor of higher case fatality. The longer term risk of death associated with diabetes was greatest in younger patients. In patients aged 65 years or younger, the hazard ratio for mortality at 5 years associated with diabetes was 1.41 (95% CI, 1.31 to 1.52) for men and 1.64 (1.50 to 1.79) for women. The risk associated with diabetes was less in patients aged 75 years or older: a hazard ratio in men 1.16 (1.10 to 1.22) and in women 1.15 (1.10 to 1.20). In the younger age group the risk associated with diabetes was significantly greater in women than in men (P=0.005 for diabetes-sex interaction). Diabetes was also a significant independent predictor of heart failure readmission, and again the risk was greatest in younger women. CONCLUSIONS: Although diabetes was associated with a lower case fatality at 30 days, by 1 year it was a significant independent predictor of higher case fatality. The risk associated with diabetes was greatest in young patients, and in young patients the risk was greatest in women. PMID- 19808298 TI - Metabolic syndrome, inflammation, and incident heart failure in the elderly: the cardiovascular health study. AB - BACKGROUND: Inflammation markers and metabolic syndrome (MetS) are associated with risk of congestive heart failure (CHF). We evaluated whether combining inflammation markers and MetS provided additive information for incident CHF and if incorporating inflammation markers to the MetS definition added prognostic information. METHODS AND RESULTS: We studied 4017 men and women > or =65 years old, without baseline CHF or diabetes, participating in the Cardiovascular Health Study, an observational study with 12.2 years follow-up and 966 cases of incident CHF. Baseline "C-reactive protein (CRP)-MetS" or "interleukin (IL)-6-MetS" were defined as presence of 3 out of 6 components, with elevated CRP (> or =3 mg/L) or IL-6 (> or =2.21 pg/mL) as a sixth component added to ATPIII criteria. Cox models adjusted for CHF risk factors and incident coronary disease were used to calculate hazard ratios for CHF. MetS and elevated inflammation markers were independently associated with CHF risk (hazard ratios, 95% CI: 1.32, 1.16 to 1.51 for MetS; 1.53, 1.34 to 1.75 for CRP; 1.37, 1.19 to 1.55 for IL-6). There was a 20% relative excess risk attributed to the combination of MetS and CRP (95% CI, 44% to 88%). CRP-MetS and IL-6-MetS definitions reclassified 18% and 13%, respectively of participants as MetS. Both CRP-MetS and IL-6-MetS increased risk of CHF by 60% compared with those without MetS. CONCLUSIONS: MetS and inflammation markers provided additive information on CHF risk in this elderly cohort. Inflammation-incorporated MetS definitions identified more participants with the same risk level as ATPIII MetS. Considering inflammation markers and MetS together may be useful in clinical and research settings. PMID- 19808300 TI - Detection of endogenous B-type natriuretic peptide at very low concentrations in patients with heart failure. AB - BACKGROUND: The myocardium secretes B-type natriuretic peptide (BNP) in response to stimuli associated with heart failure (HF). However, high immunoreactive-BNP levels in patients with HF are associated with a paradoxical lack of natriuretic response. We hypothesized that commercially available assays for immunoreactive BNP do not reflect the bioactivity of the natriuretic peptide system, because they measure both unprocessed inactive pro-BNP and mature BNP 1-32. We describe an assay for the detection of bioactive BNP 1-32 and confirm very low concentrations in plasma from HF patients. METHODS AND RESULTS: We developed a quantitative mass spectrometry immunoassay to capture endogenous BNP peptides using high affinity antibodies. Bound BNP and its truncated fragments were detected by matrix assisted laser desorption ionization-time of flight mass spectrometry based on their predicted masses. Mass spectrometry immunoassay revealed rapid in vitro degradation of BNP 1-32 in plasma, which requires plasma collection in the presence of high protease inhibitor concentrations. In 11 of 12 HF patients BNP 1-32 was detectable, ranging from 25 to 43 pg/mL. Several degraded forms of BNP were also detected at similarly low levels. In contrast, parallel measurements of immunoreactive BNP using the Biosite assay ranged from 900 to 5000 pg/mL. CONCLUSIONS: Detection of endogenous BNP 1-32 requires special preservation of plasma samples. Mass spectrometry immunoassay technology demonstrates that HF patients have low levels of BNP 1-32. Commercially available immunoreactive-BNP assays overrepresent biological activity of the natriuretic peptide system because they cannot distinguish between active and inactive forms. This observation may, in part, explain the "natriuretic paradox." PMID- 19808299 TI - Matrix metalloproteinases and their tissue inhibitors in cardiac amyloidosis: relationship to structural, functional myocardial changes and to light chain amyloid deposition. AB - BACKGROUND: Cardiac amyloidosis is characterized by amyloid infiltration resulting in extracellular matrix disruption. Amyloid cardiomyopathy due to immunoglobulin light chain protein (AL-CMP) deposition has an accelerated clinical course and a worse prognosis compared with non-light chain cardiac amyloidoses (ie, forms associated with wild-type or mutated transthyretin [TTR]). We therefore tested the hypothesis that determinants of proteolytic activity of the extracellular matrix, the matrix metalloproteinases (MMPs), and their tissue inhibitors (TIMPs) would have distinct patterns and contribute to the pathogenesis of AL-CMP versus TTR-related amyloidosis. METHODS AND RESULTS: We studied 40 patients with systemic amyloidosis: 10 AL-CMP patients, 20 patients with TTR-associated forms of cardiac amyloidosis, ie, senile systemic amyloidosis (involving wild-type TTR) or mutant TTR, and 10 patients with AL amyloidosis without cardiac involvement. Serum MMP-2 and -9, TIMP-1, -2, and -4, brain natriuretic peptide values, and echocardiography were determined. AL-CMP and TTR related amyloidosis groups had similar degrees of increased left ventricular wall thickness. However, brain natriuretic peptide, MMP-9, and TIMP-1 levels were distinctly elevated accompanied by marked diastolic dysfunction in the AL-CMP group versus no or minimal increases in the TTR-related amyloidosis group. Brain natriuretic peptide, MMPs, and TIMPs were not correlated with the degree of left ventricular wall thickness but were correlated to each other and to measures of diastolic dysfunction. Immunostaining of human endomyocardial biopsies showed diffuse expression of MMP-9 and TIMP-1 in AL-CMP and limited expression in TTR related amyloidosis hearts. CONCLUSIONS: Despite comparable left ventricular wall thickness with TTR-related cardiac amyloidosis, AL-CMP patients have higher brain natriuretic peptide, MMPs, and TIMPs, which correlated with diastolic dysfunction. These findings suggest a relationship between light chains and extracellular matrix proteolytic activation that may play an important role in the functional and clinical manifestations of AL-CMP, distinct from the other non light chain cardiac amyloidoses. PMID- 19808301 TI - Elevated afterload, neuroendocrine stimulation, and human heart failure increase BNP levels and inhibit preload-dependent SERCA upregulation. AB - BACKGROUND: In heart failure, brain-type natriuretic peptide (BNP) is elevated and the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) downregulated. We previously showed that preload-induced SERCA-upregulation is suppressed by exogenous BNP. METHODS AND RESULTS: Here we tested the hypothesis that afterload and neurohumoral activation would counterregulate preload-dependent SERCA upregulation through BNP, which finally results in decreased SERCA levels. We studied the effects of 6 hours preload, afterload, and isoproterenol stimulation on BNP and SERCA mRNA expression in rabbit and human failing muscles strips. Preload resulted in a pronounced upregulation of SERCA by 149% (isotonic versus slack, P<0.01). This upregulation was largely suppressed in afterloaded muscles (isometric versus slack: +32%; P<0.05). Similarly, presence of isoproterenol prevented SERCA upregulation in isotonic muscles. Afterload and isoproterenol resulted in a pronounced increase in BNP expression compared with slack by 225% (P<0.05) and 198% (P<0.01), respectively. Isoproterenol also increased expression of phospholamban by 84% (P<0.01). SERCA upregulation in preloaded muscles is associated with frequency-dependent potentiation of contractile force, which is absent in afterloaded muscles. In failing human myocardium, BNP expression was upregulated compared with nonfailing (+631%; P<0.05). Neither unloading nor preload or afterload induced a change in SERCA or BNP expression after 6 hours. CONCLUSIONS: Afterload and neuroendocrine stimulation increase BNP expression thereby causing inhibition of preload-dependent SERCA upregulation. In failing human myocardium, high BNP expression may underlie the loss of preload-dependent upregulation of SERCA. BNP may thus contribute to adverse myocardial remodelling in heart failure. PMID- 19808302 TI - Clinical use of phosphodiesterase-5 inhibitors in chronic heart failure. PMID- 19808303 TI - Should moderate or greater mitral regurgitation be repaired in all patients with LVEF <30%? Mitral valve repair in patients with advanced heart failure and severe functional mitral insufficiency reverses left ventricular remodeling and improves symptoms. PMID- 19808304 TI - Should moderate or greater mitral regurgitation be repaired in all patients with LVEF <30%? Surgery, mitral regurgitation, and heart failure: the valves are all repairable but the patients are not. PMID- 19808305 TI - Endocardial calcification of left atrium, tracheobronchopathia osteoplastica, and calcified aortic arch in a patient with dyspnea. PMID- 19808306 TI - Three-dimensional transesophageal echocardiogram provides real-time guidance during percutaneous paravalvular mitral repair. PMID- 19808308 TI - Innovation with experience using implantable left ventricular assist devices. PMID- 19808309 TI - Impact of center volume on outcomes of left ventricular assist device implantation as destination therapy: analysis of the Thoratec HeartMate Registry, 1998 to 2005. AB - BACKGROUND: More than 400 patients with end-stage heart failure underwent left ventricular assist device (LVAD) implantation of LVAD as destination therapy (DT) after the US Food and Drug Administration approval of DT in 2002. Because most of these patients had surgeries at hospitals that were newly accredited, we sought to examine the impact of LVAD center volume on the outcomes of DT. METHODS AND RESULTS: From July 1998 through December 2005, a total of 377 patients underwent implantation of HeartMate I LVAD as DT at 68 centers in the United States. Using data from the Thoratec DT Registry, we examined the association between LVAD center volume at the time of surgery and 1-year survival with DT. Of the studied 377 DT recipients, 53% underwent device implantation at centers that performed 9th DT implant; P=0.009). However, the DT center volume was not an independent predictor of 1-year survival with DT when adjusted for the preoperative DT Risk Score, suggesting that other factors, such as improved candidate selection, may have accounted for the institutional learning curve. CONCLUSIONS: The institutional experience with DT may have a significant impact on outcomes of this therapy. Better selection of candidates, systemic approach to surgical and postoperative care, as well as the long-term medical management most likely all contribute to these improvements. PMID- 19808310 TI - Absolute and attributable risks of heart failure incidence in relation to optimal risk factors. AB - BACKGROUND: Epidemiological studies have shown that a large proportion of coronary heart disease and stroke events are explained by borderline or elevated risk factors and that adults with optimal risk factors greatly avoid these events. The degree to which this applies to heart failure incidence is not well documented. METHODS AND RESULTS: We categorized baseline (1987-1989) risk factors in the Atherosclerosis Risk in Communities Study cohort (n=13,460, aged 45 to 64 years) into optimal, borderline, and elevated groups based on national guidelines, using a 4-factor score (blood pressure, plasma cholesterol, diabetes, and smoking) and a 5-factor score (which included body mass). Incidence of hospitalized heart failure (n=1344) was identified over a 16-year period. Only 4.9% of the cohort at baseline had all optimal risk factors based on the 4-factor score and 2.6% using the 5-factor score. Compared with participants with any elevated risk factor using the 4-factor score, the age-, sex-, and race-adjusted relative hazard for heart failure events was 0.18 (95% CI, 0.10 to 0.32) for those with all optimal risk factors and 0.35 (95% CI, 0.30 to 0.41) for those with only borderline risk factors. A population-attributable fraction estimate suggested that having at least 1 of the 4 risk factors, elevated or borderline, accounted for 77.1% of heart failure events. For the 5-factor score, that percentage was 88.8%. CONCLUSIONS: Middle-aged adults with optimal (low) risk factors have low incidence rates of heart failure, which supports redoubled efforts to prevent risk factor development in the first place. PMID- 19808311 TI - Association of multiple anthropometrics of overweight and obesity with incident heart failure: the Atherosclerosis Risk in Communities study. AB - BACKGROUND: The association of central adiposity with incident heart failure (HF) has yet to be studied in a large population-based study. METHODS AND RESULTS: The Atherosclerosis Risk in Communities study is an ongoing biracial population-based cohort of those aged 45 to 64 years from 4 US communities with 16 years' median follow-up for incident, hospitalized, or fatal HF. Waist-hip ratio, waist circumference, and body mass index (BMI) were measured at baseline (1987-1989). After exclusions, the sample size was 14 641. BMI was categorized as <25, 25 to 29.9, and >or=30 kg/m(2). Waist circumference and waist-hip ratio were divided into gender-specific tertiles. A first occurrence of International Classification of Diseases, 9th Revision, Clinical Modification, codes of HF, either hospital discharge (428.0 to 428.9; n=1451) or on a death certificate (428.0 to 428.9 or I50.0 to I50.9; n=77) was considered an HF event. Cox models were adjusted for alcohol use, smoking, age, center, and educational level. The adjusted hazard ratios for the highest category (obese) compared with the lowest were well above 1.0 for all 3 anthropometric measures (hazard ratio for 3rd versus 1st tertile of waist-hip ratio: 2.27 [1.71, 3.02] for white women; 3.24 [2.25, 4.65] for black women; 2.46 [1.95, 3.09] for white men; and 2.63 [1.90, 3.65] for black men). Hazard ratios for overweight were lower in magnitude, suggesting a graded response between body size and HF. CONCLUSIONS: Obesity and overweight, as measured by 3 different anthropometrics, were associated with incident HF in the Atherosclerosis Risk in Communities cohort. The current study does not support the superiority of waist-hip ratio and waist circumference over BMI for the prediction of incident HF. PMID- 19808312 TI - Comparative study of vasodilators in an animal model of chronic volume overload caused by severe aortic regurgitation. AB - BACKGROUND: Aortic regurgitation (AR) is a disease of chronic left ventricular (LV) volume overload. Over time, AR will lead to LV dilatation, hypertrophy, and loss of function. There is currently no medical treatment proven effective to slow the evolution of this cardiomyopathy. Vasodilators were once thought to have protective effects, but recent publications have cast some doubts about their effectiveness. We hypothesized that drugs targeting the renin-angiotensin system should be more effective than those having no direct effect on the renin angiotensin system. METHODS AND RESULTS: We designed a protocol comparing the effects of 3 vasodilators in a rat AR model (n=9 to 11 animals per group). The effects of a 6-month treatment of (1) nifedipine, (2) captopril, or (3) losartan were compared in male AR rats. Sham-operated and untreated AR animals were used as controls. Nifedipine-treated animals displayed hemodynamics, LV dilatation, hypertrophy, and loss of function similar to those of the untreated group. Both captopril and losartan were effective in improving hemodynamics, slow LV dilatation, hypertrophy, and dysfunction. Gene expression analysis confirmed the lack of effects of the nifedipine treatment at the molecular level. CONCLUSIONS: Using an animal model of severe AR, we found that vasodilators targeting the renin-angiotensin system were effective to slow the development of LV remodeling and to preserve LV function. As recently shown in the most recent human clinical trial, nifedipine was totally ineffective. Targeting the renin-angiotensin system seems a promising avenue in the treatment of this disease, and clinical trials should be carefully designed to re-evaluate the effectiveness of angiotensin I converting enzyme inhibitors or angiotensin II receptor blockers in AR. PMID- 19808313 TI - Peak cardiac power output, measured noninvasively, is a powerful predictor of outcome in chronic heart failure. AB - BACKGROUND: The cardiac output (CO) response to exercise and other invasively derived hemodynamic variables has been variably described to provide better prognostication than peak V(O(2)) in patients with chronic heart failure. Using noninvasive measurements of CO during exercise, we compared the prognostic value of peak CO and cardiac power to peak V(O(2)) in chronic heart failure patients. METHOD AND RESULTS: One hundred seventy-one consecutive patients with chronic heart failure underwent symptom limited bicycle exercise with noninvasive estimation of CO using an inert gas rebreathing method. An accurate measure of peak CO was obtained in 148 patients (85% of patients; mean age, 53+/-14 years; 80% male; left ventricular ejection fraction, 24+/-12%; ischemic etiology, 34%). Peak cardiac power was derived from the product of the peak mean arterial blood pressure and CO divided by 451. End points consisted of death, urgent heart transplant, or left ventricular assist device implantation. Duration of follow-up averaged 337+/-252 days (median, 295 days). Univariate and multivariate analysis were performed. The variables analyzed included peak V(O(2)), peak CO, peak cardiac power, V(E)/V(CO(2)) slope, and V(O(2)) at anaerobic threshold. Event free survival for the entire cohort was 83% with 5 deaths, 4 left ventricular assist device implants, and 16 urgent transplants. Peak V(O(2)) was 12.9+/-4.5 mL/kg per min, and peak cardiac power was 1.7+/-0.9 W. Peak V(O(2)), peak CO, peak cardiac power, V(E)/V(CO(2)) slope, and V(O(2)) at anaerobic threshold were predictive of outcome on univariate analysis. On multivariate analysis, peak cardiac power and peak CO were predictive of outcome with peak cardiac power being the most powerful independent predictor of outcome (P=0.01). CONCLUSIONS: Peak cardiac power, measured noninvasively, is an independent predictor of outcome that can enhance the prognostic power of peak V(O(2)) in the evaluation of patients with heart failure. PMID- 19808314 TI - Direct inotropic effects of exogenous and endogenous urotensin-II: divergent actions in failing and nonfailing human myocardium. AB - BACKGROUND: Urotensin-II (U-II) is an endogenous peptide upregulated in failing hearts. To date, insights into the myocardial actions of U-II have been obscured by its potent vasoconstrictor effects and interspecies differences in physiological responses to U-II. METHODS AND RESULTS: We examined the direct effects of exogenous U-II on in vitro contractility in nonfailing and failing human myocardial trabeculae (n=47). Rapid cooling contractures (RCC) were used to examine sarcoplasmic reticulum Ca(2+) load. In nonfailing myocardium, exogenous U II increased developed force (DF), rates of force generation and decline and RCC amplitude suggesting increased sarcoplasmic reticulum Ca(2+) load. In isolated myocyte suspensions from nonfailing hearts, U-II increased phospholamban phosphorylation. In failing myocardium, exogenous U-II reduced DF and rates of force generation and decline without a significant change in RCC amplitude in trabeculae or a change in phospholamban phosphorylation in myocytes. To examine the effects of endogenous U-II, we administered the peptidic U-II receptor antagonist (UT-A) GSK248451A to isolated trabeculae. UT-A induced a decrease in DF in nonfailing myocardium and an increase in DF in failing myocardium. UT-A increased RCC amplitude slightly in both nonfailing and failing myocardium. During ongoing UT-A, exogenous U-II had little effect on DF and RCC amplitude, confirming effective receptor blockade. CONCLUSIONS: U-II modulates contractility independent of vasoconstriction with opposite effects in failing and nonfailing hearts. Positive inotropic responses to UT-A alone suggests that increased endogenous U-II constrains contractility in failing hearts via an autocrine or paracrine mechanism. These findings support a potential therapeutic role for UT-A in heart failure. PMID- 19808315 TI - Reduced stretch-induced force response in failing human myocardium caused by impaired Na(+)-contraction coupling. AB - BACKGROUND: Stretch elicits an immediate, followed by a delayed, inotropic response in various animal models and failing human myocardium. This study aimed to characterize functional differences in the stretch response between failing and nonfailing human myocardium. METHODS AND RESULTS: Experiments were performed in muscle tissue from 86 failing and 16 nonfailing human hearts. Muscles were stretched from 88% to 98% of optimal length. Resulting immediate (Frank-Starling mechanism [FSM]) and delayed (slow-force response [SFR]) increases in twitch force were assessed before and after blockade of nitric oxide synthase, phosphatidylinositol-3-kinase, or reverse-mode Na(+)/Ca(2+) exchange. Stretch induced changes in [Na(+)](i) were measured using fluorescent indicator sodium binding benzofuran isophthalate-AM. Nitric oxide synthase isoform expression was quantified by Western blot analysis. FSM was comparable between nonfailing (227+/ 8%) and failing (222+/-9%) myocardium, whereas the additional increase during SFR (approximately 5 minutes) was larger in nonfailing myocardium (to 126+/-3% versus 119+/-2% of force of FSM, respectively; P<0.05). Basal [Na(+)](i) and stretch induced increase in [Na(+)](i) were lower in nonfailing myocardium. Inhibition of the Na(+)/H(+) exchange largely reduced the increase in [Na(+)](i) and significantly blocked the SFR. In both groups, SFR was almost completely prevented by reverse-mode Na(+)/Ca(+)-exchanger inhibition. Although neuronal and inducible nitric oxide synthase expression were significantly upregulated in failing myocardium, inhibition of nitric oxide synthase and phosphatidylinositol 3-kinase had no effect on FSM or SFR. CONCLUSIONS: These data demonstrate a Na(+) independent FSM and a Na(+)-dependent SFR in both nonfailing and failing human myocardium. The larger stretch-dependent increase in [Na(+)](i) in failing myocardium was associated with a blunted functional response, indicating impaired Na(+)-contraction coupling in the failing human heart. PMID- 19808316 TI - Heart failure in children: part I: history, etiology, and pathophysiology. PMID- 19808317 TI - Managing patients with rapid atrial fibrillation and decompensated heart failure. PMID- 19808318 TI - Atrial fibrillation and acute decompensated heart failure. PMID- 19808319 TI - Acute heart failure due to midaortic occlusion as the initial manifestation of Takayasu arteritis. PMID- 19808320 TI - An atypical presentation of endomyocardial fibrosis diagnosed by cardiac MRI. PMID- 19808321 TI - Digoxin: quo vadis? PMID- 19808322 TI - Serum cystatin C, renal filtration function, and left ventricular remodeling. PMID- 19808323 TI - Digoxin therapy does not improve outcomes in patients with advanced heart failure on contemporary medical therapy. AB - BACKGROUND: The impact of digoxin on outcomes of patients with advanced heart failure (HF) receiving optimal contemporary therapy is not known. METHODS AND RESULTS: We retrospectively reviewed data of 455 advanced HF patients referred for transplant evaluation (age, 52+/-12 years; ejection fraction, 18.3+/-8%); 227 (49.9%) were on digoxin at baseline. Primary outcome was death (n=101), urgent transplantation (n=14), or ventricular assist device implantation (n=4); secondary outcomes included HF and all-cause hospitalizations. Digoxin use was evaluated (1) in the original cohort; (2) in a propensity score-matched subset (n=322); (3) as a time-dependent covariate; and (4) after adjustment for Seattle Heart Failure Score. Patients were on optimal therapy: angiotensin-II modulation, 92.5%; beta-blockers, 91.2%; aldosterone antagonists, 45.6%; and devices, 71.0%. After a median of 27 months, 83 of 277 (36.6%) patients treated with digoxin versus 36 of 228 (15.8%) patients without digoxin met primary outcome (hazard ratio [HR], 2.28; 95% CI, 1.51 to 3.43; P<0.001). This risk persisted in the matched subset (HR, 1.73; 95% CI, 1.09 to 2.75; P=0.021) and with time-varying digoxin use (HR, 2.05; 95% CI, 1.23 to 3.41; P=0.011). Digoxin was associated with higher risk among patients in sinus rhythm compared with atrial fibrillation. Digoxin was not associated with improvement in either all-cause or HF hospitalization rates. These results were similar across sex and race and when adjusted for Seattle Heart Failure Score and renal function. CONCLUSIONS: This study suggests that digoxin therapy may be of no benefit in patients with advanced HF referred for cardiac transplantation who received optimal medical therapy. Treatment with digoxin should be used cautiously in such patients because of risk for adverse outcomes. PMID- 19808324 TI - Association of cystatin C with left ventricular structure and function: the Dallas Heart Study. AB - BACKGROUND: Cystatin C, a novel marker of renal function, has been associated with heart failure and cardiovascular mortality in older individuals. We tested the hypothesis that cystatin C is associated with preclinical cardiac structural and functional abnormalities in a younger population-based sample. METHODS AND RESULTS: The study included participants in the Dallas Heart Study (ages 30 to 65 years) who had measurements of cystatin C and cardiac MRI. The associations of cystatin C with left ventricular (LV) mass, LV end-systolic and -diastolic volumes, concentricity (LV mass/LV end-diastolic volume), LV wall thickness, and LV ejection fraction were evaluated. Cystatin C levels ranged from 0.46 to 6.55 mg/L. In univariable analyses, increasing levels of cystatin C correlated with higher LV mass, concentricity, and wall thickness (P<0.001), but not with LV end systolic volume, LV end-diastolic volume, or LV ejection fraction. After adjustment with traditional covariates and estimated glomerular filtration rate by the modification of diet in renal disease formula, log-transformed cystatin C remained independently associated with LV mass (P<0.001), concentricity (P=0.027), and wall thickness (P<0.001). These associations persisted when creatinine or estimated glomerular filtration rate by the Cockcroft-Gault formula were included in the models. CONCLUSIONS: Higher levels of cystatin C were associated with increased LV mass and a concentric LV hypertrophy phenotype. These findings were independent of potential confounding variables including standard measurements of renal function, supporting the hypothesis that cystatin C may be useful to identify individuals with preclinical structural heart abnormalities. PMID- 19808325 TI - Prevalence of left ventricular diastolic dysfunction in a general population. AB - BACKGROUND: Because the process of myocardial remodelling starts before the onset of symptoms, recent heart failure (HF) guidelines place special emphasis on the detection of subclinical left ventricular (LV) systolic and diastolic dysfunction and the timely identification of risk factors for HF. Our goal was to describe the prevalence and determinants (risk factors) of LV diastolic dysfunction in a general population and to compare the amino terminal probrain natriuretic peptide level across groups with and without diastolic dysfunction. METHODS AND RESULTS: In a randomly recruited population sample (n=539; 50.5% women; mean age, 52.5 years), we measured early and late diastolic peak velocities of mitral inflow (E and A), pulmonary vein flow by pulsed-wave Doppler, and the mitral annular velocities (Ea and Aa) at 4 sites by tissue Doppler imaging. A healthy subsample of 239 subjects (mean age, 43.7 years) provided age-specific cutoff limits for normal E/A and E/Ea ratios and the differences in duration between the mitral A and the reverse pulmonary vein flows during atrial systole (DeltaAd-ARd). The number of subjects in diastolic dysfunction groups 1 (impaired relaxation), 2 (elevated LV end-diastolic filling pressure), and 3 (elevated E/Ea and abnormally low E/A) were 53 (9.8%), 76 (14.1%), and 18 (3.4%), respectively. We used Delta(Ad or =31.9, P<0.001) and added prognostic value to ventilatory efficiency (VE/Vco(2) slope), the strongest cardiopulmonary exercise testing predictor of adverse events (chi(2)=150.7, P<0.001), in a multivariate regression. The Wasserman/Hansen prediction equation provided optimal prognostic information. CONCLUSIONS: Actual peak Vo(2) and the percent-predicted models included in this analysis all were significant predictors of adverse events. It seems that the percent-predicted peak Vo(2) value derived from the Wasserman/Hansen equations may outperform other expressions of this cardiopulmonary exercise testing variable. PMID- 19808327 TI - Pheochromocytoma-induced cardiomyopathy is modulated by the synergistic effects of cell-secreted factors. AB - BACKGROUND: Pheochromocytomas are rare tumors derived from the chromaffin cells of the adrenal medulla. Although these tumors have long been postulated to induce hypertension and cardiomyopathy through the hypersecretion of catecholamines, catecholamines alone may not fully explain the profound myocardial remodeling induced by these tumors. We sought to determine whether changes in myocardial function in pheochromocytoma-induced cardiomyopathy result solely from catecholamines secretion or from multiple pheochromocytoma-derived factors. METHODS AND RESULTS: Isolated cardiomyocytes incubated with pheochromocytoma conditioned growth media contracted at a higher frequency than cardiomyocytes incubated with norepinephrine (NE) only. Sprague-Dawley rats and black-6 mice were implanted with agarose-encapsulated pheochromocytoma (PC12) cells, dihydroxyphenylalanine decarboxylase knock-out PC12 cells deficient in NE (PC12 KO), or NE-secreting pumps. PC12 cell implantation increased left ventricular dilation by 35+/-6% and 9.6+/-1.4% and reduced left ventricular fractional shortening by 20+/-3% and 28+/-4% in rats and mice compared with animals dosed only with NE, respectively. Elimination of NE secretion in PC12-KO cells induced neither cardiac dilation (3.9%+/-1.8% increase versus control) nor changes in (1.9%+/-0.4% reduction) fractional shortening compared to controls. CONCLUSIONS: Pheochromocytomas induce a greater degree of cardiomyopathy than equivalent doses of NE, suggesting pheochromocytoma-induced cardiomyopathy is not solely mediated by NE, rather pheochromocytoma secretory factors in combination with catecholamines act synergistically to induce greater cardiac damage than catecholamines alone. PMID- 19808328 TI - Inhibition of protein kinase C-beta by ruboxistaurin preserves cardiac function and reduces extracellular matrix production in diabetic cardiomyopathy. AB - BACKGROUND: Heart failure is a common cause of morbidity and mortality in diabetic patients that frequently manifests in the absence of impaired left ventricular systolic function. In contrast to the strong evidence base for the treatment of systolic heart failure, the treatment of heart failure with preserved left ventricular function is uncertain, and therapeutic targets beyond blockade of the renin-angiotensin-aldosterone and beta-adrenergic systems are being sought. One such target is the beta-isoform of protein kinase C (PKC), implicated in both the complications of diabetes and in cardiac dysfunction in the nondiabetic setting. METHODS AND RESULTS: Using a hemodynamically validated rodent model of diabetic diastolic heart failure, the (mRen-2)27 transgenic rat, we sought to determine whether selective inhibition of PKC-beta would preserve cardiac function and reduce structural injury. Diabetic rats were randomized to receive either vehicle or the PKC-beta inhibitor, ruboxistaurin (20 mg/kg per d) and followed for 6 weeks. Compared with untreated animals, ruboxistaurin-treated diabetic rats demonstrated preserved systolic and diastolic function, as measured by the slope of preload recruitable stroke work relationship (P<0.05) and the slope of the end-diastolic pressure volume relationship (P<0.01). Collagen I deposition and cardiomyocyte hypertrophy were both reduced in diabetic animals treated with ruboxistaurin (P<0.01), as was phosphorylated-Smad2, an index of transforming growth factor-beta activity (P<0.01 for all, versus untreated diabetic rats). CONCLUSIONS: PKC-ss inhibition attenuated diastolic dysfunction, myocyte hypertrophy, and collagen deposition and preserved cardiac contractility. PKC-beta inhibition may represent a novel therapeutic strategy for the prevention of diabetes-associated cardiac dysfunction. PMID- 19808329 TI - Small animal models of heart failure: development of novel therapies, past and present. PMID- 19808330 TI - The effects of vasodilators in pulmonary hypertension: pulmonary vascular or peripheral vascular? PMID- 19808331 TI - Heart failure as a multiple hormonal deficiency syndrome. PMID- 19808332 TI - Letter by Nguyen and Doenst regarding article "Chronic glucagon-like peptide-1 infusion sustains left ventricular systolic function and prolongs survival in the spontaneously hypertensive, heart-failure prone rat". PMID- 19808334 TI - Loss of intracellular and intercellular synchrony of calcium release in systolic heart failure. PMID- 19808335 TI - Socioeconomic position and graft failure in pediatric heart transplant recipients. AB - BACKGROUND: Socioeconomic (SE) position may affect availability of resources, health-related behavior, and outcomes. We assessed whether patient SE position, determined for the block group of patient residence (average population 1000, smallest census unit with SE data), is associated with graft failure in pediatric heart transplant recipients. METHODS AND RESULTS: We used the US Census 2000 database to derive a composite SE score for the block group of residence for all patients who underwent their first heart transplant at Children's Hospital Boston between 1991 and 2005 (n=135). Cox proportional hazards models were used to determine the risk of graft failure (death or retransplant) in the lowest tertile SE group (low SE group) compared with the remaining 2 of 3 patients (controls). The 2 groups were similar with respect to age, gender, diagnosis, and year of transplant. White race was less frequent in low SE group (64% versus 90%, P=0.001). Graft failure occurred in 46 transplant recipients (40 deaths, 6 retransplant). Low SE group (hazard ratio 2.4, 95% CI 1.3 to 4.3) and nonwhite race (hazard ratio 2.7, 95% CI 1.4 to 5.2) were both associated with higher risk of graft failure. In a multivariable model controlling for diagnosis and pretransplant support, race, and low SE position (hazard ratio 2.0, 95% CI 1.0 to 3.7, P=0.04) remained associated with graft failure. Low SE position group had a higher incidence rate of graft rejection and was at a higher risk of late rejection. CONCLUSIONS: Low SE position may be an independent risk factor for graft failure in pediatric heart transplant recipients. PMID- 19808336 TI - Novel use of cardiac pacemakers in heart failure to dynamically manipulate the respiratory system through algorithmic changes in cardiac output. AB - BACKGROUND: Alternation of heart rate between 2 values using a pacemaker generates oscillations in end-tidal CO(2) (et-CO(2)). This study examined (a) whether modulating atrioventricular delay can also do this, and (b) whether more gradual variation of cardiac output can achieve comparable changes in et-CO(2) with less-sudden changes in blood pressure. METHODS AND RESULTS: We applied pacemaker fluctuations by adjusting heart rate (by 30 bpm) or atrioventricular delay (between optimal and nonoptimal values) or both, with period of 60 s in 19 heart failure patients (age 73+/-11, EF 29+/-12%). The changes in cardiac output, by either heart rate or atrioventricular delay or both, were made either as a step ("square wave") or more gradually ("sine wave"). We obtained changes in cardiac output sufficient to engender comparable oscillations in et-CO(2) (P=NS) in all 19 patients either by manipulation of heart rate (14), or by atrioventricular delay (2) or both (3). The square wave produced 191% larger and 250% more sudden changes in blood pressure than the sine wave alternations (22.4+/-11.7 versus 13.6+/-4.5 mm Hg, P<0.01 and 19.8+/-10.0 versus 7.9+/-3.2 mm Hg over 5 s, P<0.01), but peak-to-trough et-CO(2) elicited was only 45% higher (0.45+/-0.18 versus 0.31+/-0.13 kPa, P=0.01). CONCLUSIONS: This study shows that cardiac output is the key to dynamically manipulating the respiratory system with pacing sequences. When manipulating respiration by this route, a sine wave pattern may be preferable to a square wave, because it minimizes sudden blood pressure fluctuations. PMID- 19808337 TI - Circulating plasma surfactant protein type B as biological marker of alveolar capillary barrier damage in chronic heart failure. AB - BACKGROUND: Surfactant protein type B (SPB) is needed for alveolar gas exchange. SPB is increased in the plasma of patients with heart failure (HF), with a concentration that is higher when HF severity is highest. The aim of this study was to evaluate the relationship between plasma SPB and both alveolar-capillary diffusion at rest and ventilation versus carbon dioxide production during exercise. METHODS AND RESULTS: Eighty patients with chronic HF and 20 healthy controls were evaluated consecutively, but the required quality for procedures was only reached by 71 patients with HF and 19 healthy controls. Each subject underwent pulmonary function measurements, including lung diffusion for carbon monoxide and membrane diffusion capacity, and maximal cardiopulmonary exercise test. Plasma SPB was measured by immunoblotting. In patients with HF, SPB values were higher (4.5 [11.1] versus 1.6 [2.9], P=0.0006, median and 25th to 75th interquartile), whereas lung diffusion for carbon monoxide (19.7+/-4.5 versus 24.6+/-6.8 mL/mm Hg per min, P<0.0001, mean+/-SD) and membrane diffusion capacity (28.9+/-7.4 versus 38.7+/-14.8, P<0.0001) were lower. Peak oxygen consumption and ventilation/carbon dioxide production slope were 16.2+/-4.3 versus 26.8+/-6.2 mL/kg per min (P<0.0001) and 29.7+/-5.9 and 24.5+/-3.2 (P<0.0001) in HF and controls, respectively. In the HF population, univariate analysis showed a significant relationship between plasma SPB and lung diffusion for carbon monoxide, membrane diffusion capacity, peak oxygen consumption, and ventilation/carbon dioxide production slope (P<0.0001 for all). On multivariable logistic regression analysis, membrane diffusion capacity (beta, -0.54; SE, 0.018; P<0.0001), peak oxygen consumption (beta, -0.53; SE, 0.036; P=0.004), and ventilation/carbon dioxide production slope (beta, 0.25; SE, 0.026; P=0.034) were independently associated with SPB. CONCLUSIONS: Circulating plasma SPB levels are related to alveolar gas diffusion, overall exercise performance, and efficiency of ventilation showing a link between alveolar-capillary barrier damage, gas exchange abnormalities, and exercise performance in HF. PMID- 19808338 TI - Reduction in mitral regurgitation during therapy guided by measured filling pressures in the ESCAPE trial. AB - BACKGROUND: Dynamic mitral regurgitation (MR) contributes to decompensation in chronic dilated heart failure. Reduction of MR was the primary physiological end point in the ESCAPE trial, which compared acute therapy guided by jugular venous pressure, edema, and weight (CLIN) with therapy guided additionally by pulmonary artery catheters (PAC) toward pulmonary wedge pressure or=16. CONCLUSIONS: Among patients with stable coronary artery disease and preserved ejection fraction, traditional and newer factors were independently associated with increased risk of HF. Trandolopril decreased the risk of HF in these patients with preserved ejection fraction. PMID- 19808343 TI - Leukocyte count and incidence of hospitalizations due to heart failure. AB - BACKGROUND: Leukocyte concentration in blood is a classical marker of systemic inflammation. Whether high leukocyte counts are associated with incidence of heart failure (HF) is unknown. This population-based study explored whether the leukocyte concentrations were associated with incidence of hospitalizations due to HF. METHODS AND RESULTS: Leukocyte counts were measured in 16 940 men from the general population (mean age 44 years) without history of myocardial infarction or stroke. Incidence of hospitalizations due to HF (primary diagnosis) was monitored over 23 years of follow-up, in relation to quartiles of leukocytes. Subjects with myocardial infarction during follow-up were censored in the main analysis. During the follow-up, 436 men were hospitalized due to HF. Incidence of HF hospitalizations was increased in men with high leukocyte counts. After adjustments for confounding factors, the adjusted hazards ratio (HR, 95% CI) for HF hospitalization was 1.00 (reference), 1.26 (0.93 to 1.7), 1.24 (0.91 to 1.7), and 1.73 (1.3 to 2.3), respectively, for men with leukocytes in the 1st, 2nd, 3rd, and 4th (highest) quartiles (trend, P<0.001). This relationship was consistent in smokers and nonsmokers and in men with and without hypertension, respectively. CONCLUSIONS: High leukocyte counts in middle-aged men were associated with increased long-term incidence of HF hospitalizations. PMID- 19808344 TI - Multiple defects in intracellular calcium cycling in whole failing rat heart. AB - BACKGROUND: A number of defects in excitation-contraction coupling have been identified in failing mammalian hearts. The goal of this study was to measure the defects in intracellular Ca(2+) cycling in left ventricular epicardial myocytes of the whole heart in an animal model of congestive heart failure (CHF). METHODS AND RESULTS: Intracellular Ca(2+) transients were measured using confocal microscopy in whole rat hearts from age-matched Wistar-Kyoto control rats and spontaneously hypertensive rats at approximately 23 months of age. Basal Ca(2+) transients in myocytes in spontaneously hypertensive rats were smaller in amplitude and longer in duration than Wistar-Kyoto control rats. There was also greater variability in transient characteristics associated with duration between myocytes of CHF than Wistar-Kyoto controls. Approximately 21% of CHF myocytes demonstrated spontaneous Ca(2+) waves compared with very little of this activity in Wistar-Kyoto control rats. A separate population of spontaneously hypertensive rat myocytes showed Ca(2+) waves that were triggered during pacing and were absent at rest (triggered waves). Rapid pacing protocols caused Ca(2+) alternans to develop at slower heart rates in CHF. CONCLUSIONS: Epicardial cells demonstrate both serious defects and greater cell-to-cell variability in Ca(2+) cycling in CHF. The defects in Ca(2+) cycling include both spontaneous and triggered waves of Ca(2+) release, which promote triggered activity. The slowing of Ca(2+) repriming in the sarcoplasmic reticulum is probably responsible for the increased vulnerability to Ca(2+) alternans in CHF. Our results suggest that defective Ca(2+) cycling could contribute both to reduced cardiac output in CHF and to the establishment of repolarization gradients, thus creating the substrate for reentrant arrhythmias. PMID- 19808345 TI - Prevention of myofilament dysfunction by beta-blocker therapy in postinfarct remodeling. AB - BACKGROUND: Myofilament contractility of individual cardiomyocytes is depressed in remote noninfarcted myocardium and contributes to global left ventricular pump dysfunction after myocardial infarction (MI). Here, we investigated whether beta blocker therapy could restore myofilament contractility. METHODS AND RESULTS: In pigs with a MI induced by ligation of the left circumflex coronary artery, beta blocker therapy (bisoprolol, MI+beta) was initiated on the first day after MI. Remote left ventricular subendocardial biopsies were taken 3 weeks after sham or MI surgery. Isometric force was measured in single permeabilized cardiomyocytes. Maximal force (F(max)) was lower, whereas Ca(2+) sensitivity was higher in untreated MI compared with sham (both P<0.05). The difference in Ca(2+) sensitivity was abolished by treatment of cells with the beta-adrenergic kinase, protein kinase A. beta-blocker therapy partially reversed F(max) and Ca(2+) sensitivity to sham values and significantly reduced passive force. Despite the lower myofilament Ca(2+) sensitivity in MI+beta compared with untreated myocardium, the protein kinase A induced reduction in Ca(2+) sensitivity was largest in cardiomyocytes from myocardium treated with beta-blockers. Phosphorylation of beta-adrenergic target proteins (myosin binding protein C and troponin I) did not differ among groups, whereas myosin light chain 2 phosphorylation was reduced in MI, which coincided with increased expression of protein phosphatase 1. beta-blockade fully restored the latter alterations and significantly reduced expression of protein phosphatase 2a. CONCLUSIONS: beta blockade reversed myofilament dysfunction and enhanced myofilament responsiveness to protein kinase A in remote myocardium after MI. These effects likely contribute to the beneficial effects of beta-blockade on global left ventricular function after MI. PMID- 19808346 TI - Physiological replacement of T3 improves left ventricular function in an animal model of myocardial infarction-induced congestive heart failure. AB - BACKGROUND: Patients with congestive heart failure (CHF) often have low serum triiodothyronine (T(3)) concentrations. In a rodent model of myocardial infarction-induced CHF and low serum T(3), we hypothesized that replacing T(3) to euthyroid levels would improve left ventricular function without producing untoward signs of thyrotoxicosis. METHODS AND RESULTS: Adult male Sprague-Dawley rats were subjected to left anterior descending coronary artery ligation (myocardial infarction). One week post-myocardial infarction, left ventricular fractional shortening was significantly reduced to 22+/-1% in CHF animals versus 38+/-1% for sham-operated controls (P<0.001). Serum T(3) concentration was also significantly reduced (80+/-3 versus 103+/-6 ng/dL; P<0.001), in CHF animals versus Shams. At 9 weeks post-myocardial infarction, systolic function (+dP/dt max) was significantly attenuated in CHF animals (4773+/-259 versus 6310+/-267 mmHg/s; P<0.001) as well as diastolic function measured by half time to relaxation (15.9+/-1.2 versus 11.1+/-0.3 ms; P<0.001). alpha-myosin heavy chain expression was also significantly reduced by 77% (P<0.001), and beta-myosin heavy chain expression was increased by 21%. Continuous T(3) replacement was initiated 1 week post-myocardial infarction with osmotic mini-pumps (6 microg/kg/d), which returned serum T(3) concentrations to levels similar to Sham controls while resting conscious heart rate, arterial blood pressure and the incidence of arrhythmias were not different. At 9 weeks, systolic function was significantly improved by T(3) replacement (6279+/-347 mmHg/s; P<0.05) and a trend toward improved diastolic function (12.3+/-0.6 ms) was noted. T(3) replacement in CHF animals also significantly increased alpha- and reduced beta-MHC expression, (P<0.05). CONCLUSIONS: These data indicate that T(3) replacement to euthyroid levels improves systolic function and tends to improve diastolic function, potentially through changes in myocardial gene expression. PMID- 19808347 TI - Progress with genetic cardiomyopathies: screening, counseling, and testing in dilated, hypertrophic, and arrhythmogenic right ventricular dysplasia/cardiomyopathy. AB - This review focuses on the genetic cardiomyopathies: principally dilated cardiomyopathy, with salient features of hypertrophic cardiomyopathy and arrhythmogenic right ventricular dysplasia/cardiomyopathy, regarding genetic etiology, genetic testing, and genetic counseling. Enormous progress has recently been made in identifying genetic causes for each cardiomyopathy, and key phenotype and genotype information is reviewed. Clinical genetic testing is rapidly emerging with a principal rationale of identifying at-risk asymptomatic or disease-free relatives. Knowledge of a disease-causing mutation can guide clinical surveillance for disease onset, thereby enhancing preventive and treatment interventions. Genetic counseling is also indicated for patients and their family members regarding the symptoms of their cardiomyopathy, its inheritance pattern, family screening recommendations, and genetic testing options and possible results. PMID- 19808349 TI - On the control of metabolic remodeling in mitochondria of the failing heart. PMID- 19808348 TI - Large animal models of heart failure: a critical link in the translation of basic science to clinical practice. AB - Congestive heart failure (HF) is a clinical syndrome, with hallmarks of fatigue and dyspnea, that continues to be highly prevalent and morbid. Because of the growing burden of HF as the population ages, the need to develop new pharmacological treatments and therapeutic interventions is of paramount importance. Common pathophysiologic features of HF include changes in left ventricle structure, function, and neurohormonal activation. The recapitulation of the HF phenotype in large animal models can allow for the translation of basic science discoveries into clinical therapies. Models of myocardial infarction/ischemia, ischemic cardiomyopathy, ventricular pressure and volume overload, and pacing-induced dilated cardiomyopathy have been created in dogs, pigs, and sheep for the investigation of HF and potential therapies. Large animal models recapitulating the clinical HF phenotype and translating basic science to clinical applications have successfully traveled the journey from bench to bedside. Undoubtedly, large animal models of HF will continue to play a crucial role in the elucidation of biological pathways involved in HF and the development and refinement of HF therapies. PMID- 19808350 TI - Value of exercise treadmill testing in the risk stratification of patients with pulmonary hypertension. AB - BACKGROUND: The ability of the Naughton-Balke exercise treadmill test, an objective indicator of exercise capacity, to predict abnormal hemodynamics and mortality in pulmonary hypertension is unknown. METHODS AND RESULTS: We performed a cohort study of 603 patients with pulmonary hypertension from 1982 to 2006, and studied the utility of exercise treadmill test as a predictor of abnormal hemodynamics and death. We used multivariable linear regression to determine whether exercise capacity, measured in metabolic equivalents, was associated with abnormal hemodynamics, and we used a Cox proportional hazards model to determine whether decreased exercise capacity predicted death. Mean age was 50+/-15 years, 76% were women, 63% had World Health Organization category I pulmonary arterial hypertension, and 23% were World Health Organization functional classes I and II. Mean exercise capacity was 3.7+/-2.2 metabolic equivalents. Decreased exercise capacity was independently associated with elevated right atrial and mean pulmonary artery pressure, decreased cardiac index, and increased pulmonary vascular resistance. During median follow-up of 4.6 years, 36% of the patients died. Decreased exercise capacity was associated with mortality (multivariable hazard ratio, 1.18; 95% CI, 1.01 to 1.37 for each 1-metabolic equivalent decrease in exercise capacity; P=0.031; P=0.052 after adjusting for invasive hemodynamic variables). Decreased exercise capacity also predicted mortality in functional classes I-II patients, 24% of whom died (hazard ratio, 1.53; 95% CI, 1.04 to 2.26 for each 1-metabolic equivalent decrease in exercise capacity; P=0.032), although this association did not persist after adjusting for invasive hemodynamic variables (P=0.63). CONCLUSIONS: Reduced exercise capacity on exercise treadmill test is associated with worse hemodynamics and is a predictor of mortality in patients with pulmonary hypertension. PMID- 19808351 TI - Rapid Emergency Department Heart Failure Outpatients Trial (REDHOT II): a randomized controlled trial of the effect of serial B-type natriuretic peptide testing on patient management. AB - BACKGROUND: B-type natriuretic peptide is useful to diagnose heart failure. We determined whether the use of serial B-type natriuretic peptide measurements to guide treatment improves the outcome in patients with acute heart failure. METHODS AND RESULTS: We conducted a randomized controlled trial of patients with acute heart failure in 10 academic and community emergency departments. The experimental group received serial B-type natriuretic peptide testing (at 3, 6, 9, and 12 hours then daily). The control group received usual care. Our outcomes were hospital length of stay, 30-day readmission rate, and all-cause mortality. There were 219 controls and 228 experimental patients. Mean age was 64 years, 49% were women, 58% were blacks, and 34% were whites. Groups were similar in baseline characteristics. Comparing the serial testing with the control group, there was no difference in length of stay (6.5 days [95% CI, 5.2 to 7.9] versus 6.5 days [95% CI, 5.6 to 7.3]; difference, 0.1 [95% CI, -1.7 to 1.5]), in-hospital mortality (2.2% [95% CI, 0.9 to 5.0] versus controls, 3.2% [95% CI, 1.6 to 6.5]; difference, 1.0% [95% CI, -2.3 to 4.5]), 30-day mortality (3.7% [95% CI, 1.8 to 7.5] versus 5.5% [95% CI, 3.0 to 9.8]; difference, 1.8% [95% CI, -2.8 to 6.5]), or hospital revisit rate (20.2% [95% CI, 15.0 to 26.6] versus 23.7% [95% CI, 18.0 to 30.6]; difference, 3.5% [95% CI, -5.1 to 12.1]). CONCLUSIONS: In this study of 447 patients hospitalized for suspected heart failure, we were unable to demonstrate a benefit of serial testing with B-type natriuretic peptide in terms of hospital length of stay, mortality, or readmission rate. PMID- 19808352 TI - Prevalence and prognostic significance of elevated gamma-glutamyltransferase in chronic heart failure. AB - BACKGROUND: Serum gamma-glutamyltransferase (GGT) is associated with incident cardiovascular diseases and is a potential risk factor for disease mortality. We investigated the relevance of circulating GGT in chronic heart failure. METHODS AND RESULTS: From 2000 to 2007 clinical and laboratory variables of 1033 consecutive outdoor patients with heart failure were evaluated. Follow-up (mean, 34.4 months) was available in 998 patients. The end point was defined as death from any cause or heart transplantation. A forward stepwise Cox proportional hazards regression model for sex-stratified data was used. Prevalence of elevated GGT was 42.9% in men (GGT >65 U/L) and 50.2% in women (GGT >38 U/L), which was higher than for sex- and age-matched healthy subjects (18.6% in men, 19.2% in women) derived from a large historical control group. GGT was associated with severity of heart failure as assessed by New York Heart Association class, left ventricular ejection fraction, and amino-terminal pro-B-type natriuretic peptide. The end point was recorded in 302 patients. Compared with the lowest GGT quintile, sex-stratified hazard ratios for patients in the highest quintile were 2.88 (1.99 to 4.17) in the univariate model and 1.87 (1.28 to 2.74) in the adjusted model (P<0.001). Corresponding 5-year cumulative event rates were 47% and 74%, respectively. Adjusted hazard ratios for elevated GGT was 2.9 (1.64 to 5.17) for patients in New York Heart Association I/II, and 1.2 (0.75 to 2.05) for patients in New York Heart Association III/IV, respectively (P=0.003, for the GGT New York Heart Association class interaction). CONCLUSIONS: Prevalence of elevated GGT is high in patients with chronic heart failure. The GGT levels are associated with disease severity. Increased GGT is an independent predictor of death or heart transplantation. GGT may provide additional prognostic information, especially in patients with mild heart failure. PMID- 19808353 TI - Association between elevated fibrosis markers and heart failure in the elderly: the cardiovascular health study. AB - BACKGROUND: Myocardial fibrosis reflects excess collagen deposition in the extracellular left ventricular matrix, which has been associated with heart failure (HF). No studies have addressed the relation between fibrosis biomarkers and HF in the elderly. METHODS AND RESULTS: Serum fibrosis markers were measured in 880 participants of the Cardiovascular Health Study (mean age 77+/-6 years, 48% women). Participants with systolic HF (n=131, left ventricular ejection fraction <55%) and those with diastolic HF (n=179, left ventricular ejection fraction > or =55%) were compared with controls (280 with cardiovascular risk factors, and 279 healthy individuals) using a nested case-control design. Fibrosis markers included carboxyl-terminal peptide of procollagen type I, carboxyl-terminal telopeptide of collagen type I, and amino-terminal peptide of procollagen type III. Echocardiography was used to document systolic and diastolic function parameters. Analysis of variance and logistic regression analysis (per tertile odds ratios [OR]), adjusted by age, gender, race, hypertension, atrial fibrillation, coronary heart disease, baseline serum glucose, serum cystatin C, serum creatinine, C-reactive protein, any angiotensin converting enzyme inhibitor, spironolactone or any diuretic, NT-proBNP, and total bone mineral density were performed. Systolic HF was associated with significantly elevated carboxyl-terminal telopeptide of collagen type I (OR=2.6; 95% CI=1.2 to 5.7) and amino-terminal peptide of procollagen type III (OR=3.3; 95% CI=1.6 to 5.8), when adjusting for covariates. Associations of diastolic HF were significant for carboxyl-terminal telopeptide of collagen type I (OR=3.9; 95% CI=1.9 to 8.3) and amino-terminal peptide of procollagen type III (OR=2.7; 95% CI=1.4 to 5.4). HF was not associated with elevated carboxyl-terminal peptide of procollagen type I (P>0.10), and fibrosis markers did not significantly differ between HF with diastolic versus those with systolic dysfunction (P>0.10) whereas NT-proBNP mean values were higher in systolic heart failure than in diastolic heart failure (P<0.0001). CONCLUSIONS: Fibrosis markers are significantly elevated in elderly individuals with diastolic or systolic HF. These associations remained significant when adjusting for covariates relevant to the aging process. PMID- 19808354 TI - Serum levels of the interleukin-1 receptor family member ST2, cardiac structure and function, and long-term mortality in patients with acute dyspnea. AB - BACKGROUND: ST2, a biomarker of cardiomyocyte stretch, powerfully predicts poor outcomes in patients with acute dyspnea, but nothing is known about associations between soluble ST2 (sST2) and cardiac structure and function, or whether sST2 retains prognostic meaning in the context of such measures. METHODS AND RESULTS: One hundred thirty-four dyspneic patients with and without decompensated heart failure had echocardiography during index admission and vital status was ascertained at 4 years. Echocardiographic and clinical correlates of sST2 as well as independent predictors of death at 4 years were identified. sST2 correlated with left ventricular end-systolic dimensions/volumes and left ventricular ejection fraction. sST2 was inversely associated with right ventricular fractional area change (rho=-0.18; P=0.046), higher right ventricular systolic pressure (rho=0.26; P=0.005), and right ventricular hypokinesis (P<0.001) and was correlated with tissue Doppler Ea wave peak velocity, but not to other indices of diastolic function. In multivariate regression, independent predictors of sST2 included right ventricular systolic pressure (t=2.29; P=0.002), left ventricular ejection fraction (t=-2.15; P=0.05) and dimensions (end systolic, t=2.57; end diastolic, t=2.98; both P<0.05), amino-terminal pro-B-type natriuretic peptide (t=3.31; P=0.009), heart rate (t=2.59; P=0.01), and presence of jugular venous distension (t=2.00; P=0.05). In a Cox proportional hazards model that included echocardiographic results and other biomarkers, sST2 independently predicted death at 4 years (hazard ratio=2.70; P=0.003). CONCLUSIONS: Among dyspneic patients with and without acute heart failure, sST2 concentrations are associated with prevalent cardiac abnormalities on echocardiography, a more decompensated hemodynamic profile and are associated with long-term mortality, independent of echocardiographic, clinical, or other biochemical markers of risk. PMID- 19808355 TI - Prognosis on chronic dobutamine or milrinone infusions for stage D heart failure. AB - BACKGROUND: There are no published clinical trials comparing dobutamine with milrinone in outpatients with stage D heart failure on continuous inotropes. METHODS AND RESULTS: In a retrospective analysis of 112 inotrope-dependent patients with stage D heart failure who were not transplant candidates at enrollment, we investigated the relationship between choice of dobutamine or milrinone and mortality. Half the patients were on dobutamine (mean dose, 5.4+/ 2.5 microg/kg per minute) and half on milrinone (mean dose, 0.4+/-0.2 microg/kg per minute). Those on dobutamine tended to be older (63 years old versus 54 years old), male (86% versus 79%), and fewer had implantable cardioverter defibrillators (57% versus 74%). During a median follow-up time of 130 days (range, 2 to 2345 days), there were 85 deaths (76% of cohort) and 55 rehospitalizations. Use of dobutamine compared with milrinone was associated with higher all-cause mortality in an unadjusted analysis (hazard ratio [HR], 1.63; 95% CI, 1.06 to 2.52; P<0.03). However, this association was not significant after adjustment for baseline characteristics in the full cohort (N=112; HR, 0.99; 95% CI 0.5 to 1.97; P=0.98) or propensity-matched cohort (N=70; HR, 0.94; 95% CI 0.48 to 1.85; P=0.86). CONCLUSIONS: In this single-center retrospective study, there were no mortality differences between chronic intravenous dobutamine or milrinone in patients with stage D heart failure being discharged from the hospital. The high mortality in this group selected for inotrope dependence warrants careful consideration of all options and priorities for further care. PMID- 19808356 TI - Expression patterns of cardiac myofilament proteins: genomic and protein analysis of surgical myectomy tissue from patients with obstructive hypertrophic cardiomyopathy. AB - BACKGROUND: Mutations in myofilament proteins, most commonly MYBPC3-encoded myosin-binding protein C and MYH7-encoded beta-myosin heavy chain, can cause hypertrophic cardiomyopathy (HCM). Despite significant advances in structure function relationships pertaining to the cardiac sarcomere, there is limited knowledge of how a mutation leads to clinical HCM. We, therefore, set out to study expression and localization of myofilament proteins in left ventricular tissue of patients with HCM. METHODS AND RESULTS: Frozen surgical myectomy specimens from 47 patients with HCM were examined and genotyped for mutations involving 8 myofilament-encoding genes. Myofilament protein levels were quantified by Western blotting with localization graded from immunohistochemical staining of tissue sections. Overall, 25 of 47 (53%) patients had myofilament HCM, including 12 with MYBPC3-HCM and 9 with MYH7-HCM. As compared with healthy heart tissue, levels of myofilament proteins were increased in patients manifesting a mutation in either gene. Patients with a frameshift mutation predicted to truncate MYBPC3 exhibited marked disturbances in protein localization as compared with missense mutations in either MYBPC3 or MYH7. CONCLUSIONS: In this first expression study in human HCM tissue, increased myofilament protein levels in patients with either MYBPC3- or MYH7-mediated HCM suggest a poison peptide mechanism. Specifically, the mechanism of dysfunction may vary according to the genetic subgroup suggested by a distinctly abnormal distribution of myofilament proteins in patients manifesting a truncation mutation in MYBPC3. PMID- 19808357 TI - Effects of cardiac myosin isoform variation on myofilament function and crossbridge kinetics in transgenic rabbits. AB - BACKGROUND: The left ventricles of both rabbits and humans express predominantly beta-myosin heavy chain (MHC). Transgenic (TG) rabbits expressing 40% alpha-MHC are protected against tachycardia-induced cardiomyopathy, but the normal amount of alpha-MHC expressed in humans is only 5% to 7% and its functional importance is questionable. This study was undertaken to identify a myofilament-based mechanism underlying tachycardia-induced cardiomyopathy protection and to extrapolate the impact of MHC isoform variation on myofilament function in human hearts. METHODS AND RESULTS: Papillary muscle strips from TG rabbits expressing 40% (TG40) and 15% alpha-MHC (TG15) and from nontransgenic (NTG) controls expressing approximately 100% beta-MHC (NTG40 and NTG15) were demembranated and calcium activated. Myofilament tension and calcium sensitivity were similar in TGs and respective NTGs. Force-clamp measurements revealed approximately 50% higher power production in TG40 versus NTG40 (P<0.001) and approximately 20% higher power in TG15 versus NTG15 (P<0.05). A characteristic of acto-myosin crossbridge kinetics, the "dip" frequency, was significantly higher in TG40 versus NTG40 (0.70+/-0.04 versus 0.39+/-0.09 Hz, P<0.01) but not in TG15 versus NTG15. The calculated crossbridge time-on was also significantly shorter in TG40 (102.3+/-14.2 ms) versus NTG40 (175.7+/-19.7 ms) but not in TG15 versus NTG15. CONCLUSIONS: The incorporation of 40% alpha-MHC leads to greater myofilament power production and more rapid crossbridge cycling, which facilitate ejection and relengthening during short cycle intervals, and thus protect against tachycardia-induced cardiomyopathy. Our results suggest, however, that, even when compared with the virtual absence of alpha-MHC in the failing heart, the 5% to 7% alpha-MHC content of the normal human heart has little if any functional significance. PMID- 19808358 TI - Control by circulating factors of mitochondrial function and transcription cascade in heart failure: a role for endothelin-1 and angiotensin II. AB - BACKGROUND: Evidence is emerging to support the concept that the failing heart is "energy depleted" and that defects in energy metabolism are important determinants in the development and the progression of the disease. We have shown previously that depressed mitochondrial function in cardiac and skeletal muscles in chronic heart failure is linked to decreased expression of the gene encoding transcriptional proliferator-activated receptor-gamma coactivator-1alpha, the inducible regulator of mitochondrial biogenesis and its transcription cascade, leading to altered expression of mitochondrial proteins. However, oxidative capacity of the myocardium of patients treated for chronic heart failure and pathophysiological mechanisms of mitochondrial dysfunction are still largely unknown. METHODS AND RESULTS: In patients with chronic heart failure treated with angiotensin-converting enzyme inhibition, cardiac oxidative capacity, measured in saponin-permeabilized fibers, was 25% lower, and proliferator-activated receptor gamma coactivator-1alpha protein content was 34% lower compared with nonfailing controls. In a rat model of myocardial infarction, angiotensin-converting enzyme inhibition therapy was only partially able to protect cardiac mitochondrial function and transcription cascade. Expression of proliferator-activated receptor gamma coactivator-1alpha and its transcription cascade were evaluated after a 48 hour exposure of cultured adult rat ventricular myocytes to endothelin-1, angiotensin II, aldosterone, phenylephrine, or isoprenaline. Endothelin-1 (-30%) and, to a lesser degree, angiotensin II (-20%) decreased proliferator-activated receptor-gamma coactivator-1alpha mRNA content, whereas other hormones had no effect (phenylephrine) or even increased it (aldosterone, isoprenaline). CONCLUSIONS: Taken together, these results show that, despite angiotensin converting enzyme inhibition treatment, oxidative capacity is reduced in human and experimental heart failure and that endothelin-1 and angiotensin II could be involved in the downregulation of the mitochondrial transcription cascade. PMID- 19808359 TI - Cardiac-restricted overexpression of membrane type-1 matrix metalloproteinase in mice: effects on myocardial remodeling with aging. AB - BACKGROUND: The direct consequences of a persistently increased myocardial expression of the unique matrix metalloproteinase (MMP) membrane type-1 (MT1-MMP) on myocardial remodeling remained unexplored. METHODS AND RESULTS: Cardiac restricted MT1-MMPexp was constructed in mice using the full-length human MT1-MMP gene ligated to the myosin heavy chain promoter, which yielded approximately a 200% increase in MT1-MMP when compared with age/strain-matched wild-type (WT) mice. Left ventricular (LV) function and geometry was assessed by echocardiography in 3-month ("young") WT (n=32) and MT1-MMPexp (n=20) mice and compared with 14-month ("middle-aged") WT (n=58) and MT1-MMPexp (n=35) mice. LV end-diastolic volume was similar between the WT and MT1-MMPexp young groups, as was LV ejection fraction. In the middle-aged WT mice, LV end-diastolic volume and ejection fraction was similar to young WT mice. However, in the MT1-MMPexp middle aged mice, LV end-diastolic volume was approximately 43% higher and LV ejection fraction 40% lower (both P<0.05). Moreover, in the middle-aged MT1-MMPexp mice, myocardial fibrillar collagen increased by nearly 2-fold and was associated with approximately 3-fold increase in the processing of the profibrotic molecule, latency-associated transforming growth factor binding protein. In a second study, 14-day survival after myocardial infarction was significantly lower in middle aged MT1-MMPexp mice. CONCLUSIONS: Persistently increased myocardial MT1-MMP expression, in and of itself, caused LV remodeling, myocardial fibrosis, dysfunction, and reduced survival after myocardial injury. These findings suggest that MT1-MMP plays a mechanistic role in adverse remodeling within the myocardium. PMID- 19808360 TI - Donor CD4 T cells contribute to cardiac allograft vasculopathy by providing help for autoantibody production. AB - BACKGROUND: The development of autoantibody after heart transplantation is increasingly associated with poor graft outcome, but what triggers its development and whether it has a direct causative role in graft rejection is not clear. Here, we study the development of antinuclear autoantibody in an established mouse model of heart allograft vasculopathy. METHODS AND RESULTS: Humoral vascular changes, including endothelial complement staining, were present in bm12 heart grafts, explanted 50 days after transplantation. Alloantibody was not detectable, but long-lasting autoantibody responses developed in C57BL/6 recipients from the third week after transplantation. No autoantibody was generated if donor CD4 T cells were depleted before heart graft retrieval or in recipients that lacked B-cell major histocompatibility complex class II expression, indicating that humoral autoimmunity is a consequence of donor CD4 T cell allorecognition of the major histocompatibility complex class II complex on recipient autoreactive B cells. An effector role for autoantibody in graft rejection was confirmed by abrogation of humoral vascular rejection, and attenuation of vasculopathy, in B-cell deficient recipients and by development of vascular obliteration and accelerated rejection in recipients primed for autoantibody before transplantation. CONCLUSIONS: Passenger CD4 T cells within heart transplants can contribute to allograft vasculopathy by providing help to recipient B cells for autoantibody generation. PMID- 19808361 TI - Sodium retention in heart failure and cirrhosis: potential role of natriuretic doses of mineralocorticoid antagonist? AB - Patients with cirrhosis and heart failure (HF) share the pathophysiology of decreased effective arterial blood volume because of splanchnic vasodilatation in cirrhosis and decreased cardiac output in HF, with resultant stimulation of the renin-angiotensin-aldosterone system. Hyperaldosteronism plays a major role in the pathogenesis of ascites and contributes to resistance to loop diuretics. Therefore, the use of high doses of aldosterone antagonist (spironolactone up to 400 mg/day) is the main therapy to produce a negative sodium balance in cirrhotic patients with ascites. Hyperaldosteronism also has increasingly been recognized as a risk factor for myocardial and vascular fibrosis. Therefore, low-dose aldosterone antagonists are being used in patients with HF for cardioprotective action. However, the doses (25 to 50 mg/day) at which they are being used in cardiac patients as reported in the Randomized Aldactone Evaluation Study are not natriuretic. It is likely, therefore, that the mortality benefit relates primarily from their effect on cardiac and vascular fibrosis. Resistance to commonly used loop diuretics is frequently present in patients with advanced HF. In patients with decompensated HF with volume overload who are loop diuretic resistant, ultrafiltration may be the only available option. This is, however, an invasive procedure. For these patients, natriuretic doses of aldosterone antagonists (spironolactone >50 mg/day) may be a potential option. The competitive natriuretic response of aldosterone antagonists is related to activity of the renin-angiotensin-aldosterone system: the higher the renin angiotensin-aldosterone system activity, the higher the dose of aldosterone antagonist required to produce natriuresis. This article will discuss the potential use of natriuretic doses of aldosterone antagonists in patients with HF, including the potential side effect of hyperkalemia. PMID- 19808362 TI - Should we use outpatient dobutamine or milrinone? PMID- 19808363 TI - Role of the brain natriuretic peptide in heart failure management. PMID- 19808364 TI - Natriuretic Peptide measurements in managing heart failure: in theory and in practice. PMID- 19808365 TI - Unusual cause of acute pulmonary edema: unruptured aneurysm of noncoronary sinus causing severe mitral obstruction. PMID- 19808366 TI - Are multiple biomarker testing strategies ready for prime time in heart failure? PMID- 19808367 TI - Sex matters, but to what clinical avail? PMID- 19808368 TI - Prognostic value of biomarkers in heart failure: application of novel methods in the community. AB - BACKGROUND: Mortality among patients with heart failure is high. Though individual biomarkers have been investigated to determine their value in mortality risk prediction, the role of a multimarker strategy requires further evaluation. METHODS AND RESULTS: Olmsted County residents presenting with heart failure from July 2004 to September 2007 were recruited to undergo biomarker measurement. We investigated whether addition of C-reactive protein, B-type natriuretic peptide, and troponin T to a model including established risk indicators improved 1-year mortality risk prediction using the c statistic, integrated discrimination improvement, and net reclassification improvement. Among 593 participants, the mean age was 76.4 years, and 48% were men. After 1 year of follow-up, 122 (20.6%) participants had died. Patients with C-reactive protein (<11.8 mg/L), B-type natriuretic peptide (<350 pg/mL), and troponin T (< or = 0.01 ng/mL) less than the median had low 1-year mortality (3.3%), whereas those with 2 or 3 biomarkers greater than the median had markedly increased mortality (30.8% and 35.5%, respectively). The addition of 2 or more biomarkers to the model offered greater improvement in 1-year mortality risk prediction than use of a single biomarker. The combination of C-reactive protein and B-type natriuretic peptide resulted in an increase in the c statistic from 0.757 to 0.810 (P<0.001), an integrated discrimination improvement gain of 7.1% (P<0.001), and a net reclassification improvement of 22.1% (P<0.001). Use of all 3 biomarkers offered no incremental gain (integrated discrimination improvement gain 0.7% versus C-reactive protein+B-type natriuretic peptide, P=0.065). CONCLUSIONS: Biomarkers improved 1-year mortality risk prediction beyond established indicators. The use of a 2-biomarker combination was superior to a single biomarker in risk prediction, though addition of a third biomarker conferred no added benefit. PMID- 19808369 TI - The impact of donor-recipient sex matching on survival after orthotopic heart transplantation: analysis of 18 000 transplants in the modern era. AB - INTRODUCTION: Single-institution series have suggested that men receiving orthotopic heart transplantation from female donors have decreased survival. No multi-institutional series has comprehensively addressed the issue of donor and recipient sex matching for both male and female orthotopic heart transplantation recipients. METHODS AND RESULTS: We used data from the multi-institutional prospectively collected United Network for Organ Sharing open transplantation cohort to review 18 240 adult patients who received orthotopic heart transplantation from 1999 to 2007. Four donor recipient strata were identified (male donor/male recipient, N=10 750; female donor/female recipient, N=2201; male donor/female recipient, N=2121; and female donor/male recipient, N=3168). The primary end point of all cause posttransplant mortality was compared among groups using a Cox proportional hazard regression model with additional propensity adjustment. Female recipients, irrespective of donor sex, had 3.6% lower overall survival at 5 years posttransplant (P=0.003). Men who received organs from male donors had the highest cumulative survival at 5 years (74.5%). Men receiving female hearts had a 15% increase in the risk of adjusted cumulative mortality (hazard ratio, 1.15; 95% CI, 1.02 to 1.30; P=0.02). No significant increase in the relative hazard for death occurred for women receiving opposite sex donor organs (1.24; 0.92 to 1.35; P=0.31). CONCLUSIONS: The United Network for Organ Sharing data set has provided a large sample examining donor recipient sex pairing in orthotopic heart transplantation. Men receiving organs for same sex donors have significantly improved short- and long-term survival. No survival advantage was seen for women with same sex donors. PMID- 19808370 TI - Endothelin-1 is a key mediator of coronary vasoconstriction in patients with transplant coronary arteriosclerosis. AB - BACKGROUND: Transplant coronary arteriosclerosis (TCA) is the principal long-term complication in cardiac transplant recipients. The mediators responsible for vascular proliferation and vasoconstriction typical of TCA remain largely unknown. We tested whether endothelin-1 (ET-1), a potent vasoconstrictor and mitogen, contributes to the pathogenesis and manifestations of TCA. METHODS AND RESULTS: BQ-123, an ET-1 receptor-A antagonist, was infused into a coronary artery (40 nmol/min for 60 minutes) of 18 subjects, 6 + or - 4 years after transplantation. Vasomotor responses were measured in the infused artery and in a noninfused control artery in patients with (n=10) and without (n=8) advanced TCA (108 total coronary segments). Changes in diameters were compared at 15-minute intervals up to 60 minutes. Contribution of ET-1 to coronary constrictor tone was assessed by comparing vasodilation from BQ-123 with that of the maximal vasodilator nitroglycerin (200-microg intracoronary bolus). BQ-123 dilated coronary arteries of transplanted patients (8.4% at 60 minutes versus -0.4% in noninfused arteries, P<0.001). Dilation was greater for arteries with advanced TCA defined as diameter stenosis > or = 15% (dilation 15.2% with versus 0.6% without advanced TCA, P=0.004). Judged against the response to nitroglycerin, ET 1 accounted for 53.2% of coronary tone in advanced TCA but only 12.9% without advanced TCA. CONCLUSIONS: This study shows for the first time in humans that ET 1 is an important mediator of coronary vasoconstriction in TCA and accounts for >50% of the increased vasomotor tone. Therapeutic targeting of ET-1 may retard the development of TCA. PMID- 19808371 TI - A pilot trial to assess potential effects of selective intracoronary bone marrow derived progenitor cell infusion in patients with nonischemic dilated cardiomyopathy: final 1-year results of the transplantation of progenitor cells and functional regeneration enhancement pilot trial in patients with nonischemic dilated cardiomyopathy. AB - BACKGROUND: Intracoronary administration of bone marrow-derived progenitor cells (BMC) was shown to improve coronary microvascular function in ischemic heart disease. Because coronary microvascular dysfunction is implicated in the pathogenesis and prognosis of nonischemic dilated cardiomyopathy (DCM), we investigated the effects of intracoronary BMC administration in patients with DCM. METHODS AND RESULTS: Intracoronary infusion of BMC was performed in 33 patients with DCM by using an over-the-wire balloon catheter. Left ventricular contractility at baseline and after 3 months was assessed by analysis of left ventricular angiograms. Coronary hemodynamics were determined by intracoronary Doppler wire measurements. After 3 months, regional wall motion of the target area (contractility from -1.08 + or - 0.39 to -0.97 + or - 0.47 SD/chord, P=0.029) and global left ventricular ejection fraction (from 30.2 + or - 10.9 to 33.4 + or - 11.5%, P<0.001) were improved. Increase of regional contractile function was directly related to the functionality of the infused cells as measured by their colony-forming capacity. Minimal vascular resistance index was significantly reduced in the BMC-treated vessel after 3 months (from 1.53 + or - 0.63 to 1.32 + or -0.61 mm Hg x s/cm; P=0.002, n=24), whereas no changes were observed in the reference vessel (from 1.60 + or - 0.45 to 1.49 + or - 0.45 mm Hg x s/cm; P=0.133, n=13). Twelve months after BMC infusion, N-terminal prohormone brain natriuretic peptide (NT-proBNP) serum levels were decreased, suggesting a beneficial effect on left ventricular remodeling processes (from 1610 + or - 993 to 1473 + or - 1147 pg/mL; P=0.038 for logNT-proBNP, n=26). CONCLUSIONS: Intracoronary administration of BMC seems to be associated with improvements in cardiac contractile and microvascular function in patients with DCM. Thus, randomized blinded studies are warranted to evaluate potential clinical benefits of intracoronary BMC administration in patients with DCM. PMID- 19808372 TI - The effectiveness of collaborative medicine reviews in delaying time to next hospitalization for patients with heart failure in the practice setting: results of a cohort study. AB - BACKGROUND: Randomized controlled trials have demonstrated that collaborative medication reviews can improve outcomes for patients with heart failure. We aimed to determine whether these results translated into Australian practice, where collaborative reviews are nationally funded. METHODS AND RESULTS: This retrospective cohort study using administrative claims data included veterans 65 years and older receiving bisoprolol, carvedilol, or metoprolol succinate for which prescribing physicians indicated treatment was for heart failure. We compared those exposed to a general practitioner-pharmacist collaborative home medication review with those who did not receive the service. The service includes physician referral, a home visit by an accredited pharmacist to identify medication-related problems, and a pharmacist report with follow-up undertaken by the physician. Kaplan-Meier analyses and Cox proportional hazards models were used to compare time until next hospitalization for heart failure between the exposed and unexposed groups. There were 273 veterans exposed to a home medicines review and 5444 unexposed patients. Average age in both groups was 81.6 years (no significant difference). The median number of comorbidities was 8 in the exposed group and 7 in the unexposed (P<0.0001). Unadjusted results showed a 37% reduction in rate of hospitalization for heart failure at any time (hazard ratio, 0.63; 95% CI, 0.44 to 0.89). Adjusted results showed a 45% reduction (hazard ratio, 0.55; 95% CI, 0.39 to 0.77) among those who had received a home medicines review compared with the unexposed patients. CONCLUSIONS: Medicines review in the practice setting is effective in delaying time to next hospitalization for heart failure in those treated with heart failure medicines. PMID- 19808373 TI - Lipoprotein-associated phospholipase A(2) and risk of congestive heart failure in older adults: the Cardiovascular Health Study. AB - BACKGROUND: Inflammation may be a causative factor in congestive heart failure (CHF). Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) is an inflammation marker associated with vascular risk. One previous study showed an association of Lp-PLA(2) activity with CHF risk, but there were only 94 CHF cases and Lp-PLA(2) antigen, which is available clinically in the United States, was not measured. METHODS AND RESULTS: We measured baseline Lp-PLA(2) antigen and activity in 3991 men and women without baseline CHF or cardiovascular disease who were participating in the Cardiovascular Health Study, a prospective observational study of adults 65 years or older. Cox proportional hazards models adjusted for age, sex, clinic site, race, low-density and high-density lipoprotein cholesterol, body mass index, systolic and diastolic blood pressure, hypertension, smoking status, pack-years, and diabetes were used to calculate hazard ratios and 95% CIs for incident CHF. Further models adjusted for coronary disease events during follow-up and C-reactive protein. Eight hundred twenty-nine participants developed CHF during 12.1 years. Adjusted hazard ratios for CHF with Lp-PLA(2) in the fourth compared with the first quartile were 1.44 (95% CI, 1.16 to 1.79) for Lp-PLA(2) antigen and 1.06 (95% CI, 0.84 to 1.32) for activity. Adjustment for incident coronary disease attenuated the hazard ratio for Lp PLA(2) antigen to 1.26 (95% CI, 1.02 to 1.57), adjustment for C-reactive protein had minimal impact. CONCLUSIONS: Lp-PLA(2) antigen was associated with risk of future CHF in older people, independent of CHF and coronary risk factors, and partly mediated by coronary disease events. Further clinical and basic research is needed to better understand the role of Lp-PLA(2) in CHF. PMID- 19808374 TI - Moderate exercise training improves survival and ventricular remodeling in an animal model of left ventricular volume overload. AB - BACKGROUND: Exercise training has beneficial effects in patients with heart failure, although there is still no clear evidence that it may impact on their survival. There are no data regarding the effects of exercise in subjects with chronic left ventricular (LV) volume overload. Using a rat model of severe aortic valve regurgitation (AR), we studied the effects of long-term exercise training on survival, development of heart failure, and LV myocardial remodeling. METHODS AND RESULTS: One hundred sixty male adult rats were divided in 3 groups: sham sedentary (n=40), AR sedentary (n=80), and AR trained (n=40). Training consisted in treadmill running for up to 30 minutes, 5 times per week for 9 months, at a maximal speed of 20 m/minute. All sham-operated animals survived the entire course of the protocol. After 9 months, 65% of trained animals were alive compared with 46% of sedentary ones (P=0.05). Ejection fractions remained in the normal range (all above 60%) and LV masses between AR groups were similar. There was significantly less LV fibrosis in the trained group and lower LV filling pressures and improved echocardiographic diastolic parameters. Heart rate variability was also improved by exercise. CONCLUSIONS: Our data show that moderate endurance training is safe, does not increase the rate of developing heart failure, and most importantly, improves survival in this animal model of chronic LV volume overload. Exercise improved LV diastolic function, heart rate variability, and reduced myocardial fibrosis. PMID- 19808375 TI - Loss of angiotensin-converting enzyme 2 accelerates maladaptive left ventricular remodeling in response to myocardial infarction. AB - BACKGROUND: Angiotensin-converting enzyme 2 (ACE2) is a monocarboxypeptidase that metabolizes Ang II into Ang 1-7, thereby functioning as a negative regulator of the renin-angiotensin system. We hypothesized that ACE2 deficiency may compromise the cardiac response to myocardial infarction (MI). METHODS AND RESULTS: In response to MI (induced by left anterior descending artery ligation), there was a persistent increase in ACE2 protein in the infarct zone in wild-type mice, whereas loss of ACE2 enhanced the susceptibility to MI, with increased mortality, infarct expansion, and adverse ventricular remodeling characterized by ventricular dilation and systolic dysfunction. In ACE2-deficient hearts, elevated myocardial levels of Ang II and decreased levels of Ang 1-7 in the infarct related zone was associated with increased production of reactive oxygen species. ACE2 deficiency leads to increased matrix metalloproteinase (MMP) 2 and MMP9 levels with MMP2 activation in the infarct and peri-infarct regions, as well as increased gelatinase activity leading to a disrupted extracellular matrix structure after MI. Loss of ACE2 also leads to increased neutrophilic infiltration in the infarct and peri-infarct regions, resulting in upregulation of inflammatory cytokines, interferon-gamma, interleukin-6, and the chemokine, monocyte chemoattractant protein-1, as well as increased phosphorylation of ERK1/2 and JNK1/2 signaling pathways. Treatment of Ace2(-)(/y)-MI mice with irbesartan, an AT1 receptor blocker, reduced nicotinamide-adenine dinucleotide phosphate oxidase activity, infarct size, MMP activation, and myocardial inflammation, ultimately resulting in improved post-MI ventricular function. CONCLUSIONS: We conclude that loss of ACE2 facilitates adverse post-MI ventricular remodeling by potentiation of Ang II effects by means of the AT1 receptors, and supplementing ACE2 can be a potential therapy for ischemic heart disease. PMID- 19808376 TI - Functional analysis of a unique troponin c mutation, GLY159ASP, that causes familial dilated cardiomyopathy, studied in explanted heart muscle. AB - BACKGROUND: Familial dilated cardiomyopathy can be caused by mutations in the proteins of the muscle thin filament. In vitro, these mutations decrease Ca(2+) sensitivity and cross-bridge turnover rate, but the mutations have not been investigated in human tissue. We studied the Ca(2+)-regulatory properties of myocytes and troponin extracted from the explanted heart of a patient with inherited dilated cardiomyopathy due to the cTnC G159D mutation. METHODS AND RESULTS: Mass spectroscopy showed that the mutant cTnC was expressed approximately equimolar with wild-type cTnC. Contraction was compared in skinned ventricular myocytes from the cTnC G159D patient and nonfailing donor heart. Maximal Ca(2+)-activated force was similar in cTnC G159D and donor myocytes, but the Ca(2+) sensitivity of cTnC G159D myocytes was higher (EC(50) G159D/donor=0.60). Thin filaments reconstituted with skeletal muscle actin and human cardiac tropomyosin and troponin were studied by in vitro motility assay. Thin filaments containing the mutation had a higher Ca(2+) sensitivity (EC(50) G159D/donor=0.55 + or - 0.13), whereas the maximally activated sliding speed was unaltered. In addition, the cTnC G159D mutation blunted the change in Ca(2+) sensitivity when TnI was dephosphorylated. With wild-type troponin, Ca(2+) sensitivity was increased (EC(50) P/unP=4.7 + or - 1.9) but not with cTnC G159D troponin (EC(50) P/unP=1.2 + or - 0.1). CONCLUSIONS: We propose that uncoupling of the relationship between phosphorylation and Ca(2+) sensitivity could be the cause of the dilated cardiomyopathy phenotype. The differences between these data and previous in vitro results show that native phosphorylation of troponin I and troponin T and other posttranslational modifications of sarcomeric proteins strongly influence the functional effects of a mutation. PMID- 19808377 TI - Plasma urocortin 1 in human heart failure. AB - BACKGROUND: The urocortins are emerging as potentially important contributors to neurohumoral regulation of the circulation with recent reports attributing a powerful array of hemodynamic, renal, and neurohumoral effects to the urocortins in cardiac failure. These peptides also seem to have cardioprotective effects in the setting of ischemia-reperfusion. Little is known concerning the plasma concentrations of the urocortins in health and disease. We have investigated plasma urocortin 1 as a potential diagnostic marker of heart failure and documented its relationships to symptoms, measures of cardiac function, and concurrent levels of other circulating neurohormones. METHODS AND RESULTS: In 299 patients with recent onset dyspnea or peripheral edema presenting to primary care, plasma urocortin 1 and other vasoactive hormones were assayed, and echocardiography was performed. Heart failure was present in 74 patients (25%) according to predefined diagnostic criteria. Urocortin 1 levels were increased in patients with heart failure and were related to functional class, clinical signs of heart failure, echocardiographic indicators of left ventricular dimensions and function, plasma creatinine, and concurrent circulating levels of plasma natriuretic peptides, adrenomedullin, and endothelin 1. CONCLUSIONS: Plasma urocortin 1 is elevated in heart failure (in proportion to the degree of cardiac dysfunction) in concert with the generalized neurohormonal activation seen in this condition. Urocortin levels predict heart failure independent of age, history of previous myocardial infarction, diabetes, hypertension, fractional shortening, and N-terminal prohormone brain natriuretic peptide levels. PMID- 19808378 TI - Impairment of diastolic function by lack of frequency-dependent myofilament desensitization rabbit right ventricular hypertrophy. AB - BACKGROUND: Ventricular hypertrophy is a physiological response to pressure overload that, if left untreated, can ultimately result in ventricular dysfunction, including diastolic dysfunction. The aim of this study was to test the hypothesis that frequency-dependent myofilament desensitization, a physiological response of healthy myocardium, is altered in hypertrophied myocardium. METHODS AND RESULTS: New Zealand white rabbits underwent a pulmonary artery banding procedure to induce pressure overload. After 10 weeks, the animals were euthanized, hearts removed, and suitable trabeculae harvested from the free wall of the right ventricle. Twitch contractions, calibrated bis-fura-2 calcium transients, and myofilament calcium sensitivity (potassium contractures) were measured at frequencies of 1, 2, 3, and 4 Hz. The force frequency response, relaxation frequency response, and calcium frequency relationships were significantly blunted, and diastolic tension significantly increased with frequency in the pulmonary artery banding rabbits compared with sham-operated animals. Myofilament calcium sensitivity was virtually identical at 1 Hz in the treatment versus sham group (pCa 6.11 + or - 0.03 versus 6.11 + or - 0.06), but the frequency-dependent desensitization that takes place in the sham group (DeltapCa 0.14 + or - 0.06, P<0.05) was not observed in the pulmonary artery banding animals (DeltapCa 0.02 + or - 0.05). Analysis of myofilament protein phosphorylation revealed that the normally observed frequency-dependent phosphorylation of troponin-I is lost in pulmonary artery banding rabbits. CONCLUSIONS: The frequency-dependent myofilament desensitization is significantly impaired in right ventricular hypertrophy and contributes to the frequency dependent elevation of diastolic tension in hypertrophy. PMID- 19808379 TI - Characterization of an extensive transverse tubular network in sheep atrial myocytes and its depletion in heart failure. AB - BACKGROUND: In ventricular myocytes, the majority of structures that couple excitation to the systolic rise of Ca(2+) are located at the transverse tubular (t-tubule) membrane. In the failing ventricle, disorganization of t-tubules disrupts excitation contraction coupling. The t-tubule membrane is virtually absent in the atria of small mammals resulting in spatiotemporally distinct profiles of intracellular Ca(2+) release on stimulation in atrial and ventricular cells. The aims of this study were to determine (i) whether atrial myocytes from a large mammal (sheep) possess t-tubules, (ii) whether these are functionally important, and (iii) whether they are disrupted in heart failure. METHODS AND RESULTS: Sheep left atrial myocytes were stained with di-4-ANEPPS. Nearly all control cells had an extensive t-tubule network resulting in each voxel in the cell being nearer to a membrane (sarcolemma or t-tubule) than would otherwise be the case. T-tubules decrease the distance of 50% of voxels from a membrane from 3.35 + or - 0.15 to 0.88 + or- 0.04 microm. During depolarization, intracellular Ca(2+) rises simultaneously at the cell periphery and center. In heart failure induced by rapid ventricular pacing, there was an almost complete loss of atrial t-tubules. The distance of 50% of voxels from a membrane increased to 2.04 + or - 0.08 microm, and there was a loss of early Ca(2+) release from the cell center. CONCLUSIONS: Sheep atrial myocytes possess a substantial t-tubule network that synchronizes the systolic Ca(2+) transient. In heart failure, this network is markedly disrupted. This may play an important role in changes of atrial function in heart failure. PMID- 19808380 TI - Heart failure in children: part II: diagnosis, treatment, and future directions. PMID- 19808381 TI - Treatment of congestion in congestive heart failure: ultrafiltration is the only rational initial treatment of volume overload in decompensated heart failure. PMID- 19808382 TI - Ultrafiltration should not replace diuretics for the initial treatment of acute decompensated heart failure. PMID- 19808383 TI - Heart failure and cardiac involvement as isolated manifestation of familial form of transthyretin amyloidosis resulting from Val30Met mutation with no clinical signs of polyneuropathy. PMID- 19808384 TI - Overdrive pacing suppresses ectopy and minimizes left ventricular assist device suction events. PMID- 19808385 TI - Videofluoroscopic modified barium swallow study for premature ventricular complexes-associated dysphagia. PMID- 19808386 TI - Arrhythmia and Electrophysiology: the eagle can land. PMID- 19808387 TI - The substrate maintaining persistent atrial fibrillation. PMID- 19808388 TI - Randomized evaluation of right atrial ablation after left atrial ablation of complex fractionated atrial electrograms for long-lasting persistent atrial fibrillation. AB - BACKGROUND: With electrogram-guided radiofrequency ablation (RFA) of long-lasting persistent atrial fibrillation (AF), the best results have been reported when complex fractionated electrograms (CFAEs) in both the left (LA) and right (RA) atria were targeted. However, many studies have reported excellent outcomes from RFA of long-lasting persistent AF with the use of other ablation strategies that were limited to the LA. The incremental value of RFA of RA CFAEs is yet to be defined. METHODS AND RESULTS: In 85 patients with long-lasting persistent AF (age=59+/-10 years), RFA was directed at CFAEs in the LA and coronary sinus until AF terminated (19) or all identified LA CFAEs were eliminated. Sixty-six patients who remained in AF were randomly assigned to cardioversion and no further RFA (n=33) or to RFA of RA CFAEs (n=33). RA sites consisted of the crista terminalis (69%), septum (38%), superior vena cava (28%), coronary sinus ostium (22%), and the base of the appendage (31%). AF terminated in 1 (3%) of 33 patients during RA RFA. At 17+/-6 months after a single ablation procedure, 74% of the patients in whom AF terminated during LA RFA were in sinus rhythm. Rates of freedom from AF were similar in the patients randomized to no RFA in the RA (24%) and those randomized to RFA of RA CFAEs (30%, P=0.8). The ablation procedure was repeated in 26 patients (31%) for AF (n=22) or atrial flutter (n=4). At 16+/-7 months after the final procedure, 89% of the patients in whom AF terminated during LA RFA were in sinus rhythm. Among the randomized patients, the proportion of patients who remained in sinus rhythm was similar in patients who did not undergo RFA of RA CFAEs (52%) and those who did (58%, P=0.6). CONCLUSIONS: After RFA of CFAEs in the LA and coronary sinus, ablation of CFAEs in the RA provides little or no increment in efficacy among patients with long-lasting persistent AF. PMID- 19808389 TI - Atrial tachycardia after ablation of persistent atrial fibrillation: identification of the critical isthmus with a combination of multielectrode activation mapping and targeted entrainment mapping. AB - BACKGROUND: Atrial tachycardia (AT) that develops after ablation of atrial fibrillation often poses a more difficult clinical situation than the index arrhythmia. This study details the use of an impedance-based electroanatomic mapping system (Ensite NavX) in concert with a specialized multielectrode mapping catheter for rapid, high-density atrial mapping. In this study, this activation mapping was combined with entrainment mapping to eliminate ATs developing late after atrial fibrillation ablation. METHODS AND RESULTS: All study patients developed AT after ablation for atrial fibrillation. The approach to AT ablation consisted of 4 steps: use of a 20-pole penta-array catheter to map the chamber rapidly during the rhythm of interest, analysis of the patterns of atrial activation to identify wave fronts of electric propagation, targeted entrainment at putative channels, and catheter ablation at these "isthmuses." All ablations were performed with irrigated radiofrequency ablation catheters. Forty-one ATs were identified in 17 patients (2.4+/-1.6 ATs per patient). Using the multielectrode catheter in conjunction with the Ensite NavX system, we created activation maps of 33 of 41 ATs (81%) (mean cycle length, 284+/-71 seconds) with a mean of 365+/-108 points per map and an average mapping time of 8+/-3 minutes. Of the 33 mapped ATs, 7 terminated either spontaneously or during entrainment maneuvers. Radiofrequency energy was used to attempt ablation of 26 ATs; 25 of 26 of the ATs (96%) were terminated successfully by ablation or catheter pressure. CONCLUSIONS: This study demonstrates a strategy for rapidly defining and eliminating the scar-related ATs typically encountered after ablation of atrial fibrillation. PMID- 19808390 TI - Ventricular tachycardia originating from the posterior papillary muscle in the left ventricle: a distinct clinical syndrome. AB - BACKGROUND: Several distinct forms of focal ventricular tachycardia (VT) from the left ventricle (LV) have been described. We report a new syndrome of VT arising from the base of the posterior papillary muscle in the LV. METHODS AND RESULTS: Among 290 consecutive patients who underwent ablation for VT or symptomatic premature ventricular complexes (PVCs) based on a focal mechanism, 7 patients were found to have an ablation site at the base of the posterior papillary muscle in the LV. All patients had normal LV systolic function and a normal baseline electrocardiogram. The electrocardiogram during VT or PVCs demonstrated a right bundle-branch block and superior-axis QRS morphology in all patients. VT was not inducible by programmed atrial or ventricular stimulation. In 2 patients with sustained VT, overdrive pacing neither terminated VT nor demonstrated any criterion for transient entrainment. Activation mapping localized the earliest site of activation to the base of the posterior papillary muscle in all patients. When Purkinje potentials were recorded at the site of successful ablation, these potentials preceded local ventricular muscle potentials during sinus rhythm. During VT or PVCs, however, the ventricular muscle potential always preceded the Purkinje potentials. After recurrence of VT or PVCs with standard radiofrequency ablation, irrigated ablation was successful in eliminating the arrhythmia in all patients. Over a mean follow-up period of 9 months, all patients have been free of PVCs and VT. CONCLUSIONS: We present a distinct syndrome of VT arising from the base of the posterior papillary muscle in the LV by a nonreentrant mechanism. Ablation can be challenging, and irrigated ablation may be necessary for long term success. PMID- 19808391 TI - Mechanisms and utility of discrete great arterial potentials in the ablation of outflow tract ventricular arrhythmias. AB - BACKGROUND: Outflow tract ventricular tachycardia originating above the semilunar valves has been reported in a small number of studies. Discrete potentials in the great arteries (above the semilunar valves) have been rarely described in patients undergoing electrophysiology evaluation and radiofrequency ablation for ventricular arrhythmias. The mechanisms of these discrete potentials in the great arteries and the utility of such potentials in guiding radiofrequency ablation are unknown. METHODS AND RESULTS: Twelve patients with outflow tract ventricular arrhythmia originating above the semilunar valves with discrete arterial potentials were studied. The clinical characteristics, properties of the arterial potentials, electrophysiological evaluation and ablation, and short- and long term outcomes were reviewed. Of the twelve patients, 8 (67%) were women. The patients' average age was 41+/-14 years. The average ejection fraction was 0.52+/ 0.16 (range: 0.16 to 0.75). Contact mapping in the great artery demonstrated discrete near-field electrograms that were separate from far-field ventricular electrograms in all patients (8 above the pulmonary valve and in 4 the aortic valve). One or more of the following electrophysiological characteristics, supportive of an arrhythmogenic substrate, were observed in 10 of 12 patients: (1) A fixed or reproducibly variable pattern of discrete potential-ventricular arrhythmia relationship was present at baseline or during pacing; (2) the discrete potential-ventricular electrogram relationship during sinus rhythm was the reverse of that during the ventricular arrhythmia; (3) during sustained ventricular tachycardia, spontaneous variation of the ventricular (V-V) cycle length was preceded by a similar variation of arterial spike potential-spike potential cycle length; and (4) ablation guided by the discrete arterial potential successfully eliminated the clinical arrhythmia. Ablation was successful in these patients. In the remaining 2 patients, the potentials were believed to be bystanders. Over 10+/-4 months (range: 5 to 32 months) of follow up, there have been no recurrences of the premature ventricular complex or ventricular arrhythmia. CONCLUSIONS: Discrete potentials are present in the great arteries of a select group of patients with outflow tract ventricular tachycardia originating above the semilunar valves. When an arrhythmogenic relationship can be demonstrated, discrete potentials are useful in guiding ablation within the great vessels, despite significant anatomic complexity. PMID- 19808392 TI - Distinguishing properties of cells from the myocardial sleeves of the pulmonary veins: a comparison of normal and abnormal pacemakers. AB - BACKGROUND: A common source of arrhythmogenic spontaneous activity instigating atrial fibrillation is the myocardial tissue, or sleeves, at the base of the pulmonary veins. This study compared the properties of cells from the myocardial sleeves of the pulmonary veins (PV(m)) with cells from the normal cardiac pacemaker (the sinoatrial node) and regions of the atria. Our objective was to identify key features of these cells that predispose them to becoming the focus of cardiac arrhythmias. METHODS AND RESULTS: Single cells were isolated from samples of rabbit PV(m), central and peripheral sinoatrial node, crista terminalis, and left and right atria. Detailed morphology of cells was assessed and intracellular calcium concentrations measured with the use of Fluo-3. Cells from the PV(m) were smaller than atrial cells and showed large elevations in diastolic calcium during activation at physiological rates, a feature the PV(m) cells shared with cells from the sinoatrial node. Unstimulated spontaneous activity was observed in a minority of cells from the PV(m), but numerous cells from this region showed spontaneous activity for a brief period immediately subsequent to stimulation at physiological rates. This was not observed in atrial cells. Assessment of calcium removal pathways showed sarcolemmal calcium extrusion in cells from the PV(m) to have a high reliance on "slow" extrusion pathways to maintain intracellular calcium homeostasis because of a low expression of sodium-calcium exchanger. CONCLUSIONS: We conclude that cells from the PV(m) share some features with cells from the sinoatrial node but also have distinctly unique features that predispose them to the development of spontaneous activity. PMID- 19808393 TI - Forty years of invasive clinical electrophysiology: 1967-2007. PMID- 19808394 TI - Cardiac pacing: from biological to electronic ... to biological? AB - The prevention and treatment of life-threatening bradyarrhythmias have been revolutionized in the last half century by electronic pacemakers. Because this represents a palliative therapy, attempts have begun to effect a cure with the novel tools of gene and cell therapy. Over time, the strategies used have coalesced to focus on achieving a stable and autonomically responsive cardiac rhythm in a setting that ultimately would require no implanted hardware. In this report, we review the history of the disease process being treated, approaches now in progress, and the demands that must be met if biological therapies are to be successful. PMID- 19808395 TI - Atrial remodeling and atrial fibrillation: mechanisms and implications. PMID- 19808396 TI - Atrial fibrillation in heart failure: steady progress but still a long way to go. PMID- 19808397 TI - Dilated cardiomyopathy due to sodium channel dysfunction: what is the connection? PMID- 19808398 TI - Molecular and clinical characterization of a novel SCN5A mutation associated with atrioventricular block and dilated cardiomyopathy. AB - BACKGROUND: Increased susceptibility to dilated cardiomyopathy has been observed in patients carrying mutations in the SCN5A gene, but the underlying mechanism remains unclear. In this study, we identified and characterized, both in vitro and clinically, an SCN5A mutation associated with familial progressive atrioventricular block of adult onset and dilated cardiomyopathy in a Chinese family. METHODS AND RESULTS: Among 32 family members, 5 were initially diagnosed with atrioventricular block after age 30; 4 were studied, 3 of whom later developed dilated cardiomyopathy. We found a heterozygous single-nucleotide mutation resulting in an amino acid substitution (A1180V) in all studied patients and in 6 other younger unaffected members but not in 200 control chromosomes. When expressed with the beta1 subunit, the mutated channels exhibited a -4.5-mV shift of inactivation with slower recovery leading to a rate-dependent Na(+) current reduction and a moderate increase in late Na(+) current. Clinical study revealed that although QRS duration decreased with increasing heart rate in noncarrier family members, this change was blunted in unaffected carriers whose ECG and heart function were normal. Resting corrected QT interval of unaffected carriers was significantly longer than that of noncarriers, even though it was still within the normal range. CONCLUSIONS: A1180V expresses a mild Na(+) channel phenotype in vitro and a corresponding clinical phenotype in unaffected mutation carriers, implying that A1180V caused structural heart disease in affected carriers by disturbing Na(+) influx and, hence, cellular Na(+) homeostasis. The high penetrance of A1180V suggests this phenotype as a high risk factor for dilated cardiomyopathy with preceding atrioventricular block. PMID- 19808399 TI - Calcium-handling abnormalities underlying atrial arrhythmogenesis and contractile dysfunction in dogs with congestive heart failure. AB - BACKGROUND: Congestive heart failure (CHF) is a common cause of atrial fibrillation. Focal sources of unknown mechanism have been described in CHF related atrial fibrillation. The authors hypothesized that abnormal calcium (Ca(2+)) handling contributes to the CHF-related atrial arrhythmogenic substrate. METHODS AND RESULTS: CHF was induced in dogs by ventricular tachypacing (240 bpm x2 weeks). Cellular Ca(2+)-handling properties and expression/phosphorylation status of key Ca(2+) handling and myofilament proteins were assessed in control and CHF atria. CHF decreased cell shortening but increased left atrial diastolic intracellular Ca(2+) concentration ([Ca(2+)](i)), [Ca(2+)](i) transient amplitude, and sarcoplasmic reticulum (SR) Ca(2+) load (caffeine-induced [Ca(2+)](i) release). SR Ca(2+) overload was associated with spontaneous Ca(2+) transient events and triggered ectopic activity, which was suppressed by the inhibition of SR Ca(2+) release (ryanodine) or Na(+)/Ca(2+) exchange. Mechanisms underlying abnormal SR Ca(2+) handling were then studied. CHF increased atrial action potential duration and action potential voltage clamp showed that CHF-like action potentials enhance Ca(2+)(i) loading. CHF increased calmodulin-dependent protein kinase II phosphorylation of phospholamban by 120%, potentially enhancing SR Ca(2+) uptake by reducing phospholamban inhibition of SR Ca(2+) ATPase, but it did not affect phosphorylation of SR Ca(2+)-release channels (RyR2). Total RyR2 and calsequestrin (main SR Ca(2+)-binding protein) expression were significantly reduced, by 65% and 15%, potentially contributing to SR dysfunction. CHF decreased expression of total and protein kinase A-phosphorylated myosin-binding protein C (a key contractile filament regulator) by 27% and 74%, potentially accounting for decreased contractility despite increased Ca(2+) transients. Complex phosphorylation changes were explained by enhanced calmodulin-dependent protein kinase IIdelta expression and function and type-1 protein-phosphatase activity but downregulated regulatory protein kinase A subunits. CONCLUSIONS: CHF causes profound changes in Ca(2+)-handling and -regulatory proteins that produce atrial fibrillation-promoting atrial cardiomyocyte Ca(2+)-handling abnormalities, arrhythmogenic triggered activity, and contractile dysfunction. PMID- 19808400 TI - Altered sympathetic nervous reactivity and norepinephrine transporter expression in patients with postural tachycardia syndrome. AB - BACKGROUND: Clinical observations in patients with postural tachycardia syndrome (POTS) suggest abnormal sympathetic nervous system activity and a dysfunction of the norepinephrine (NE) transporter (NET). METHODS AND RESULTS: We examined sympathetic nervous system responses to head-up tilt by combining NE plasma kinetics measurements and muscle sympathetic nerve activity recordings and by quantifying NET protein content in peripheral sympathetic nerves in patients with POTS compared with that in controls. POTS patients had an elevated heart rate during supine rest (81+/-2 bpm versus 66+/-2 bpm in healthy subjects [HS], P<0.01). Head-up tilt to 40 degrees induced a greater rise in heart rate in patients with POTS (+24+/-4 bpm versus +13+/-2 bpm in HS, P<0.001). During rest in the supine position, muscle sympathetic nerve activity, arterial NE concentration, and whole-body NE spillover to plasma were similar in both groups. Muscle sympathetic nerve activity response to head-up tilt was greater in the POTS group (+29+/-3 bursts/min in patients with POTS and +13+/-2 bursts/min in HS, P<0.001), but the NE spillover rise was similar in both groups (51% in the POTS subjects and 50% in the HS). Western blot analysis of NET protein extracted from forearm vein biopsies in patients with POTS and HS demonstrated a decrease in the expression of NET protein in patients with POTS. CONCLUSIONS: Patients with POTS exhibit a decrease in NET protein in their peripheral sympathetic nerves. Paradoxically, whole-body NE spillover to plasma during rest in the supine position and in response to head-up tilt is not altered despite excessive nerve firing rate in response to the head-up tilt. PMID- 19808401 TI - Three-dimensional ultrasound for image-guided mapping and intervention: methods, quantitative validation, and clinical feasibility of a novel multimodality image mapping system. AB - BACKGROUND: Multiple factors create discrepancies between electroanatomic maps and merged, preacquired computed tomographic images used in guiding atrial fibrillation ablation. Therefore, a Carto-based 3D ultrasound image system (Biosense Webster Inc) was validated in an animal model and tested in 15 atrial fibrillation patients. METHODS AND RESULTS: Twelve dogs underwent evaluation using a newly developed Carto-based 3D ultrasound system. After fiducial clip markers were percutaneously implanted at critical locations in each cardiac chamber, 3D ultrasound geometries, derived from a family of 2D intracardiac echocardiographic images, were constructed. Point-source error of 3D ultrasound derived geometries, assessed by actual real-time 2D intracardiac echocardiographic clip sites, was 2.1+/-1.1 mm for atrial and 2.4+/-1.2 mm for ventricular sites. These errors were significantly less than the variance on CartoMerge computed tomographic images (atria: 3.3+/-1.6 mm; ventricles: 4.8+/ 2.0 mm; P<0.001 for both). Target ablation at each clip, guided only by 3D ultrasound-derived geometry, resulted in lesions within 1.1+/-1.1 mm of the actual clips. Pulmonary vein ablation guided by 3D ultrasound-derived geometry resulted in circumferential ablative lesions. Mapping in 15 patients produced modestly smaller 3D ultrasound versus electroanatomic map left atrial volumes (98+/-24 cm(3) versus 109+/-25 cm(3), P<0.05). Three-dimensional ultrasound guided pulmonary vein isolation and linear ablation in these patients were successfully performed with confirmation of pulmonary vein entrance/exit block. CONCLUSIONS: These data demonstrate that 3D ultrasound images seamlessly yield anatomically accurate chamber geometries. Image volumes from the ultrasound system are more accurate than possible with CartoMerge computed tomographic imaging. This clinical study also demonstrates the initial feasibility of this guidance system for ablation in patients with atrial fibrillation. PMID- 19808402 TI - Atrial arrhythmias after single-ring isolation of the posterior left atrium and pulmonary veins for atrial fibrillation: mechanisms and management. AB - BACKGROUND: Single-ring isolation of the posterior left atrium is feasible, but the incidence and mechanisms of postprocedural arrhythmias have not been described in detail. METHODS AND RESULTS: The first 100 consecutive patients (58.8+/-11.2 years old, 80 male) who underwent single-ring isolation for atrial fibrillation (66 intermittent, 18 persistent, 16 long-standing persistent) were followed up for 9.1+/-4.5 months. Recurrences were diagnosed by clinical symptoms and Holter monitoring. Patients with recurrences of sustained atrial arrhythmia >3 months after the procedure were offered a repeat procedure and were studied to determine the mechanisms of recurrence. Forty-six patients (46%) experienced sustained postprocedural atrial arrhythmias (35 had atrial fibrillation, and 34 had atrial flutter). Of these, 34 required a second procedure 7.0+/-3.1 months after their initial procedure. Reconnection of the posterior left atrium was seen in all patients with atrial fibrillation. Atrial flutter was most commonly due to mitral isthmus-dependent macroreentry (n=8, cycle length 368+/-116 ms) or macroreentry through 2 gaps in the ring of lesions (n=6, cycle length 328+/-115 ms). Posterior left atrium reisolation was achieved at the second procedure in all patients. Atrial flutter was successfully ablated and rendered noninducible in all patients. Six months after their last procedure, the Kaplan-Meier estimate of freedom from recurrence for all 100 patients was 81+/-5%. CONCLUSIONS: Atrial fibrillation and atrial flutter recurrence is common after single-ring isolation. Reconnection of the posterior left atrium and macroreentry are the common mechanisms. Repeat ablation results in satisfactory short-term outcomes. PMID- 19808403 TI - Ventricular pump function and pacing: physiological and clinical integration. PMID- 19808404 TI - Left ventricular outflow tract tachycardia with preferential conduction and multiple exits. PMID- 19808405 TI - Ablation of atrial tachycardia from a giant left superior vena cava using integration with computed tomographic imaging. PMID- 19808406 TI - Paroxysmal atrioventricular block induced by a single ventricular premature beat in the absence of overt atrioventricular conduction system disease. PMID- 19808407 TI - Ventricular tachycardia ablation: moving beyond treatment of last resort. PMID- 19808408 TI - Limitations of esophageal temperature-monitoring to prevent esophageal injury during atrial fibrillation ablation. PMID- 19808409 TI - Ventricular tachycardia ablation: evolution of patients and procedures over 8 years. AB - BACKGROUND: Evolving management of coronary artery disease, heart failure, and the use of implantable cardioverter-defibrillators impacts the characteristics of patients with recurrent ventricular tachycardia (VT). We investigated the substrate, procedure, and outcome evolution of all patients referred for VT ablation during the past 8 years. METHODS AND RESULTS: From 1999 to 2006, 493 consecutive patients (358 male, 57+/-16 years) underwent 623 VT ablations: 131 had no structural heart disease (SHD), 213 had ischemic cardiomyopathies (ICMP), and 149 had nonischemic cardiomyopathies (NICMP). Although the main substrate is ICMP, the proportion of NICMP has increased from 27% to 35% (P=0.06) from 1999 2002 to the 2003-2006. The procedure abolished or modified inducible VTs in > or =75% of patients in all groups, but abolition of all monomorphic VTs was achieved in 125 (83%) patients without SHD, 180 (65%) with ICMP, and 99 (51%) with NICMP (P<0.0001). During a mean follow-up of 3.3+/-2.4 years, no deaths occurred in patients without SHD, but 75 patients (35%) with ICMP and 26 patients (17%) with NICMP died after a median of 13 months. Multivariate Cox regression analysis found that age, ejection fraction, and need for preprocedural mechanical hemodynamic support predicted mortality. CONCLUSIONS: The substrate causing VT in patients requiring ablation is evolving and determines the long-term outcome. In the setting of a normal heart, VT ablation is associated with a low risk of subsequent mortality, with no deaths occurring during a mean follow-up of >3 years. In contrast, in patients with SHD and recurrent VT, VT ablation can be helpful to suppress drug refractory VT, but long-term mortality remains significant. PMID- 19808410 TI - Esophageal injury and temperature monitoring during atrial fibrillation ablation. AB - BACKGROUND: It is common practice to empirically limit the radiofrequency (RF) power when ablating the posterior left atrium during atrial fibrillation ablation to avoid thermal injury to the esophagus. The objective of this study was to determine whether RF energy delivery limited by luminal esophageal temperature (LET) monitoring is associated with a reduction in esophageal injury compared with a strategy of RF power limitation alone. METHODS AND RESULTS: Eighty-one consecutive patients who underwent atrial fibrillation ablation followed by esophageal endoscopy were included in this observational study. All patients underwent extraostial electric pulmonary vein isolation by using an electroanatomic mapping system and irrigated RF ablation. All RF applications on the posterior left atrium were limited to 35 W. A commercially available, single thermocouple esophageal probe was used to monitor LET in a subset of patients (n=67). In these cases, applications were promptly interrupted when LET was > or =38.5 degrees C; further applications were performed at reduced power to obtain a LET < 38.5 degrees C. Esophageal endoscopy was performed 1 to 3 days after the procedure. Ablation-related esophageal ulcerations were identified in 9 of 81 (11%) patients. All patients were asymptomatic. Of these 81 patients, LET monitoring during ablation occurred in 67 (83%) of patients. Esophageal injury was observed more frequently (36% versus 6%, P<0.006) in the group without LET monitoring. CONCLUSIONS: These data suggest that LET monitoring may be associated with a reduction in esophageal injury compared with power limitation alone. PMID- 19808411 TI - Radiofrequency puncture of the fossa ovalis for resistant transseptal access. AB - BACKGROUND: Transseptal puncture with a conventional mechanical technique can fail because of a resistant interatrial septum. We evaluated the efficacy and safety of a new method to cross-resistant septae by transmitting radiofrequency (RF) energy through the transseptal needle. METHODS AND RESULTS: Among 269 consecutive transseptal punctures, 13 (5%) were unsuccessful in 12 different patients (11 men aged 52+/-12 years) using the conventional Brockenbrough technique. All 12 patients had previously undergone at least 1 transseptal catheterization. The needle position in relation to the fossa ovalis was assessed by fluoroscopy in orthogonal views and was confirmed with contrast injection and by visualizing the characteristic "tenting" of the fossa ovalis. Before using RF energy, there were a median of 6 unsuccessful attempts to perforate the septum conventionally, with 1 pericardial puncture (with a nonsignificant effusion). RF transseptal puncture was then performed by delivering unipolar RF with manual contact between the ablation catheter and the proximal extremity of the needle at the patient's groin. RF transseptal puncture was achieved at the first attempt in all patients within a median of 1 second (interquartile range, 1 to 4) and without any complication. The only parameter predictive of a septum resistant to conventional puncture was the total number of transseptal catheterizations (3.2+/ 1 versus 1.8+/-1, P<0.001). CONCLUSIONS: Transmission of RF energy from the ablation catheter up to the tip of the transseptal needle provides an easy and safe method for piercing the fossa ovalis when the conventional approach fails because of a resistant septum. PMID- 19808412 TI - Atrial fibrillation begets atrial fibrillation: autonomic mechanism for atrial electrical remodeling induced by short-term rapid atrial pacing. AB - BACKGROUND: The mechanism(s) for acute changes in electrophysiological properties of the atria during rapid pacing induced atrial fibrillation (AF) is not completely understood. We sought to evaluate the contribution of the intrinsic cardiac autonomic nervous system in acute atrial electrical remodeling and AF induced by 6-hour rapid atrial pacing. METHODS AND RESULTS: Continuous rapid pacing (1200 bpm, 2x threshold [TH]) was performed at the left atrial appendage. Group 1 (n=7) underwent 6-hour pacing immediately followed by ganglionated plexi (GP) ablation; group 2 (n=7) underwent GP ablation immediately followed by 6-hour pacing; and group 3 (n=4) underwent administration of autonomic blockers, atropine (1 mg/kg), and propranolol (0.6 mg/kg) immediately followed by 6-hour pacing. The effective refractory period (ERP) and window of vulnerability (WOV, in milliseconds), ie, the difference between the longest and the shortest coupling interval of the premature stimulus that induced AF, were measured at 2xTH and 10xTH at the left atrium, right atrium, and pulmonary veins every hour before and after GP ablation or autonomic blockade. In group 1, ERP was markedly shortened in the first 2 hours and then stabilized both at 2xTH and 10xTH; however, WOV was progressively widened throughout the 6-hour period. After GP ablation, ERP was significantly longer than before ablation and AF could not be induced (WOV=0) at either 2xTH or 10xTH. In groups 2 and 3, rapid atrial pacing failed to shorten the ERP. AF could not be induced in 6 of 7 dogs in group 2 and all 4 dogs in group 3 during the 6-hour pacing period. CONCLUSIONS: The intrinsic cardiac autonomic nervous system plays a crucial role in the acute stages of atrial electrical remodeling induced by rapid atrial pacing. PMID- 19808413 TI - Genotyping has a minor role in selecting therapy for congenital long-QT syndromes at present. PMID- 19808414 TI - Publicly reporting implantable cardioverter defibrillator outcomes: grading the report card. PMID- 19808415 TI - Further insight into the technique and outcomes of "curative" catheter ablation of atrial fibrillation. PMID- 19808416 TI - Sudden death and defibrillators in transposition of the great arteries with intra atrial baffles: a multicenter study. AB - BACKGROUND: Transposition of the great arteries with intra-atrial baffle repair is among the congenital heart defects at highest risk of sudden death. Little is known about mechanisms of sudden death and the role of implantable cardioverter defibrillators. METHODS AND RESULTS: We conducted a multicenter cohort study in patients with transposition of the great arteries to determine actuarial rates of implantable cardioverter defibrillator shocks, identify risk factors, assess underlying arrhythmias, and characterize complications. Overall, 37 patients (age, 28.0+/-7.6 years; 89.2% male) were enrolled from 7 sites. Implantable cardioverter defibrillators were implanted for primary prevention in 23 (62.1%) patients and secondary prevention in 14 patients (37.8%). Annual rates of appropriate shocks were 0.5% and 6.0% in primary and secondary prevention, respectively (P=0.0366). Independent predictors were a secondary prevention indication (hazard ratio, 18.0; P=0.0341) and lack of beta-blockers (hazard ratio, 16.7; P=0.0301). In patients with appropriate shocks, intracardiac electrograms documented supraventricular tachycardia preceding or coexisting with ventricular tachycardia in 50%. No patient with inducible ventricular tachycardia received an appropriate shock in comparison with 37.5% of noninducible patients (P=0.0429). Inappropriate shocks occurred in 6.6% per year, more so in patients of lesser weight (hazard ratio, 0.91 per kg; P=0.0168). Additionally, 14 patients (37.8%) experienced complications: 5 (13.5%) acute, 1 (2.7%) late generator related, and 12 (32.4%) late lead related. CONCLUSIONS: In patients with transposition of the great arteries, high rates of appropriate shocks are noted in secondary but not primary prevention. Supraventricular arrhythmias may be implicated in the etiology of ventricular tachyarrhythmias; beta-blockers seem protective, and inducible ventricular tachycardia does not seem to predict future events. Inappropriate shocks and late lead-related complications are common. PMID- 19808417 TI - Fragmented wide QRS on a 12-lead ECG: a sign of myocardial scar and poor prognosis. AB - BACKGROUND: Fragmented QRS (duration <120 ms) on a 12-lead ECG represents myocardial scar in patients with coronary artery disease. However, the significance of fragmented QRS has not been defined in the presence of a wide QRS (wQRS; duration >or=120 ms). We postulate that fragmented wQRS (f-wQRS) due to bundle branch block, premature ventricular complexes, or paced rhythms (f-pQRS) signify myocardial scar and higher mortality. METHODS AND RESULTS: Patients who underwent cardiac evaluation with nuclear stress imaging or cardiac catheterization and had wQRS (bundle branch block, premature ventricular complex, or pQRS) were studied. f-wQRS was defined by the presence of >2 notches on the R wave or the S wave and had to be present in >or=2 contiguous inferior (II, III, aVF), lateral (I, aVL, V(6)) or anterior (V(1) to V(5)) leads. ECG analyses of 879 patients (age, 66.7+/-11.4 years; male, 97%; mean follow-up, 29+/-18 months) with bundle branch block (n=310), premature ventricular complex (n=301), and pQRS (n=268) revealed f-wQRS in 415 (47.2%) patients. Myocardial scar was present in 440 (50%) patients. The sensitivity, specificity, positive predictive value, and negative predictive value of f-wQRS for myocardial scar were 86.8%, 92.5%, 92.0%, and 87.5%, respectively. The sensitivity and specificity for diagnosing myocardial scar were 88.6% and 94.4%, 81.4% and 88.4%, and 89.8% and 95.7% for f bundle branch block, f-premature ventricular complex, and f-pQRS, respectively. f wQRS was associated with mortality after adjusting for age, ejection fraction, and diabetes (P=0.017). CONCLUSIONS: f-wQRS on a standard 12-lead ECG is a moderately sensitive and highly specific sign for myocardial scar in patients with known or suspected coronary artery disease. f-wQRS is also an independent predictor of mortality. PMID- 19808418 TI - Long-term clinical results of 2 different ablation strategies in patients with paroxysmal and persistent atrial fibrillation. AB - BACKGROUND: Data regarding the long-term efficacy of atrial fibrillation (AF) ablation are still lacking. METHODS AND RESULTS: Two hundred four consecutive patients symptomatic for paroxysmal or persistent/permanent AF were randomly assigned to 2 different ablation schemes: pulmonary vein isolation (PVI) and PVI plus left linear lesions (LL). Primary end point was to assess the maintenance of sinus rhythm (SR) after procedures 1 and 2 in the absence of antiarrhythmic drugs in a long-term follow-up of at least 3 years. Paroxysmal AF- With a single procedure at 12-month follow-up, 46% of patients treated with PVI maintained SR, whereas at 3-year follow-up, 29% were in SR; using the "PVI plus LL" at the 12 month follow-up, 57% of patients were in SR, whereas at the 3-year follow-up, 53% remained in SR. After a second procedure, the long-term overall success rate without antiarrhythmic drugs was 62% with PVI and 85% with PVI plus LL. Persistent/Permanent AF- With a single procedure at the 12-month follow-up, 27% of patients treated with PVI were in SR, whereas at the 3-year follow-up, 19% maintained SR; using the PVI plus LL with a single procedure at the 12-month follow-up 45% of patients were in SR, whereas at the 3-year follow-up, 41% remained in SR. After a second procedure, the long-term overall success rate without antiarrhythmic drugs was 39% with PVI and 75% with PVI plus LL. CONCLUSIONS: A long-term follow-up of AF ablation shows that short-term results cannot be considered permanent because AF recurrences are still present after the first year especially in patients who have had "PVI" strategy. PVI isolation plus LL is superior to the PVI strategy in maintaining SR without antiarrhythmic drugs after procedures 1 and 2 both in paroxysmal and persistent AF. PMID- 19808419 TI - Nodoventricular accessory pathways in PRKAG2-dependent familial preexcitation syndrome reveal a disorder in cardiac development. AB - BACKGROUND: Familial preexcitation syndrome is linked to mutations in PRKAG2. Previous studies on the R302Q mutation have provided evidence for a remarkably high proportion of otherwise rare accessory pathways with atrioventricular (AV) node-like conduction properties (Mahaim fibers). Yet, histopathologic proof is still lacking. We aimed to provide such proof. METHODS AND RESULTS: We retrospectively studied the medical records of 17 members of a 5-generation family. Five subjects died prematurely. The R302Q mutation was found in 8 living subjects and 2 deceased subjects (obligate carriers). Cardiac hypertrophy was found in 7 mutation carriers. ECGs compatible with preexcitation were found in 13 subjects and AV block at varying degrees in 5 subjects. All mutation carriers had electrocardiographic evidence of preexcitation, AV block, or both. Three individuals had high-grade AV block with preexcited conducted beats. Electrophysiological studies in 3 individuals revealed bypasses with AV node-like properties. Histopathologic studies of 1 suddenly deceased mutation carrier revealed concentric hypertrophy of the left ventricle with extensive myocardial disarray associated with slight interstitial fibrosis but no lysosomal-bound glycogen. Moreover, there were 3 small nodoventricular tracts (Mahaim fibers) passing through the central fibrous body and connecting the AV node with the working myocardium of the interventricular septum. CONCLUSIONS: Preexcitation associated with the R302Q mutation in PRKAG2 is associated with Mahaim fibers. These findings support the novel insight that PRKAG2 may be involved in the development of the cardiac conduction system. PMID- 19808420 TI - In a swine model, chest compressions cause ventricular capture and, by means of a long-short sequence, ventricular fibrillation. AB - BACKGROUND: During resuscitation, fibrillation often recurs. In swine, we studied refibrillation after long-duration ventricular fibrillation, investigating an association with chest compressions (CCs). METHODS AND RESULTS: In protocol A, 47 episodes of long-duration ventricular fibrillation lasting at least 2.5 minutes were induced in 8 animals. After defibrillation, CCs were required for 35 episodes and delivered with a pneumatic device (Lucas cardiopulmonary resuscitation). In 9 episodes, refibrillation occurred within 2 seconds of CC initiation (group 1) and in 26 episodes, CCs were delivered without refibrillation (group 2). From the ECG and intracardiac electrodes, the RR interval preceding CCs, the shortest cycle length during the first 2 CCs (short), and the preceding cycle length (long) were measured. A similar study was conducted in 3 more animals without intracardiac catheters (protocol B). In protocol A, the mean RR before CC was 665+/-292 ms in group 1 and 769+/-316 in group 2. CCs stimulated ventricular beats in all 35 episodes. The short and long intervals were shorter in group 1 (215+/-31 and 552+/-210 ms) than in group 2 (402+/-153 and 699+/-147 ms) (P=0.009 and P=0.04, respectively). The prematurity index (short/RR) was lower in group 1 than in group 2 (0.35+/-0.09 vs 0.58+/ 0.21; P<0.01). A short interval <231 ms predicted refibrillation with 88% sensitivity and 91% specificity. In protocol B, CCs were required in 11 episodes, causing ventricular stimulation in all of them and ventricular fibrillation within the first 2 CCs in 3. CONCLUSIONS: Under some conditions, CC during resuscitation can stimulate the ventricles and initiate ventricular fibrillation by a long-short sequence. PMID- 19808421 TI - Therapeutic strategies for long-QT syndrome: does the molecular substrate matter? PMID- 19808422 TI - Should patients with congenital heart disease and a systemic ventricular ejection fraction less than 30% undergo prophylactic implantation of an ICD? Patients with congenital heart disease and a systemic ventricular ejection fraction less than 30% should undergo prophylactic implantation of an implantable cardioverter defibrillator. PMID- 19808423 TI - Should patients with congenital heart disease and a systemic ventricular ejection fraction less than 30% undergo prophylactic implantation of an ICD? Implantable cardioverter defibrillator implantation guidelines based solely on left ventricular ejection fraction do not apply to adults with congenital heart disease. PMID- 19808424 TI - Cardiac myotonic dystrophy mimicking arrhythmogenic right ventricular cardiomyopathy in a young sudden cardiac death victim. PMID- 19808425 TI - Slower heart rates for healthy hearts: time to redefine tachycardia? PMID- 19808426 TI - Evolving strategies in catheter ablation of long-standing atrial fibrillation. PMID- 19808427 TI - ECG quantification of myocardial scar in cardiomyopathy patients with or without conduction defects: correlation with cardiac magnetic resonance and arrhythmogenesis. AB - BACKGROUND: Myocardial scarring from infarction or nonischemic fibrosis forms an arrhythmogenic substrate. The Selvester QRS score has been extensively validated for estimating myocardial infarction scar size in the absence of ECG confounders, but has not been tested to quantify scar in patients with hypertrophy, bundle branch/fascicular blocks, or nonischemic cardiomyopathy. We assessed the hypotheses that (1) QRS scores (modified for each ECG confounder) correctly identify and quantify scar in ischemic and nonischemic patients when compared with the reference standard of cardiac magnetic resonance using late-gadolinium enhancement, and (2) QRS-estimated scar size predicts inducible sustained monomorphic ventricular tachycardia during electrophysiological testing. METHODS AND RESULTS: One hundred sixty-two patients with left ventricular ejection fraction < or =35% (95 ischemic, 67 nonischemic) received 12-lead ECG and cardiac magnetic resonance using late-gadolinium enhancement before implantable cardioverter defibrillator placement for primary prevention of sudden cardiac death. QRS scores correctly diagnosed cardiovascular magnetic resonance scar presence with receiver operating characteristics area under the curve of 0.91 and correlation for scar quantification of r=0.74 (P<0.0001) for all patients. Performance within hypertrophy, conduction defect, and nonischemic subgroups ranged from area under the curve of 0.81 to 0.94 and r=0.60 to 0.80 (P<0.001 for all). Among the 137 patients undergoing electrophysiological or device testing, each 3-point QRS-score increase (9% left ventricular scarring) was associated with an odds ratio for inducing monomorphic ventricular tachycardia of 2.2 (95% CI, 1.5 to 3.2; P<0.001) for all patients, 1.7 (1.0 to 2.7, P=0.04) for ischemics, and 2.2 (1.0 to 5.0, P=0.05) for nonischemics. CONCLUSIONS: QRS scores identify and quantify scar in ischemic and nonischemic cardiomyopathy patients despite ECG confounders. Higher QRS-estimated scar size is associated with increased arrhythmogenesis and warrants further study as a risk-stratifying tool. PMID- 19808428 TI - Incidence of atrial fibrillation in relation to changing heart rate over time in hypertensive patients: the LIFE study. AB - BACKGROUND: Onset of atrial fibrillation (AF) has been linked to changes in autonomic tone, with increasing heart rate (HR) immediately before AF onset in some patients suggesting a possible role of acute increases in sympathetic activity in AF onset. Although losartan therapy and decreasing ECG left ventricular hypertrophy are associated with decreased AF incidence, the relationship of HR changes over time to development of AF has not been examined. METHODS AND RESULTS: HR was evaluated in 8828 hypertensive patients without AF by history or on baseline ECG in the Losartan Intervention for End Point Reduction in Hypertension (LIFE) study. Patients were treated with losartan- or atenolol based regimens and followed with serial ECGs annually which were used to determine HR and ECG left ventricular hypertrophy by Cornell product and Sokolow Lyon voltage criteria. During mean follow-up of 4.7+/-1.1 years, new-onset AF occurred in 701 patients (7.9%). Patients with new AF had smaller decreases in HR to last in-treatment ECG or last ECG before AF (-2.7+/-13.5 versus -5.2+/-12.5 bpm), whether on losartan- (-0.4+/-13.5 versus -2.2+/-11.7 bpm) or atenolol-based treatment (-5.3+/-12.8 versus -8.3+/-12.6 bpm, all P<0.001). In univariate Cox analyses, higher HR on in-treatment ECGs was associated with an increased risk of new-onset AF, with a 15% greater risk of AF for every 10 bpm higher HR (95% CI 8% to 22%). In alternative analyses, persistence or development of a HR> or =84 (upper quintile of baseline HR) was associated with a 46% greater risk of developing AF (95% CI 19% to 80%). After adjusting for treatment with losartan versus atenolol, baseline risk factors for AF, baseline and in-treatment systolic and diastolic pressure and the known predictive value of baseline and in treatment ECG left ventricular hypertrophy for new AF, higher in-treatment HR remained strongly associated with new AF with a 19% higher risk for every 10 bpm higher HR (95% CI 10% to 28%) or a 61% increased rate of AF in patients with persistence or development of a HR> or =84 (95% CI 27% to 104%, all P<0.001). CONCLUSIONS: Higher in-treatment HR on serial ECGs is associated with an increased likelihood of new-onset AF, independent of treatment modality, blood pressure lowering, and regression of ECG left ventricular hypertrophy in patients with essential hypertension. PMID- 19808429 TI - Chronic atrial fibrillation is a biatrial arrhythmia: data from catheter ablation of chronic atrial fibrillation aiming arrhythmia termination using a sequential ablation approach. AB - BACKGROUND: Termination of chronic atrial fibrillation (CAF) can be achieved by catheter ablation using a stepwise approach. However, there are limited data on the contribution of the right atrium to the CAF process. Furthermore, the prognostic value of CAF termination remains unclear. METHODS AND RESULTS: Eighty eight patients (61+/-10 years of age) underwent de novo ablation of CAF in 2006 at our institution. The ablation procedure was performed sequentially in the following order: pulmonary vein isolation, defragmentation of the left atrium, coronary sinus, and right atrium. Attempted procedural end point was termination of CAF. Consecutive arrhythmias occurring after AF termination were mapped, and ablation was attempted. AF termination was achieved in 68 (77%) patients: in 37 (55%) patients it occurred in the left atrium, in 18 (26%) patients in the right atrium, and in 13 (19%) patients in the coronary sinus. In 54 patients, at least one redo was performed (total number of procedures: 154). After the first redo, another 30 patients were in sinus rhythm (total 63), 8 patients were in atrial tachycardia (AT), and 17 patients were in AF. Another 11 patients underwent a second redo. After a mean follow-up of 20+/-4 months, 71 (81%) patients were in sinus rhythm, 1 (1%) patient was in AT, and 16 (18%) patients were in AF. Patients with CAF termination had predominantly ATs as recurrent arrhythmias (83%), whereas those without mainly presented with recurrent CAF (85%). The overall success rate in patients with CAF termination was 95% compared with 5% of patients without CAF termination in 2 procedures (n=12). In almost all redo procedures attributable to AT, at least 1 AT during redo was documented previously. CONCLUSIONS: AF termination is a prognostic important end point of catheter ablation for CAF. Termination of AF was achieved in both atria and the coronary sinus, suggesting a biatrial substrate of CAF. Subsequent arrhythmias often recur during follow-up and, therefore, should be targeted for ablation. PMID- 19808430 TI - Novel contact force sensor incorporated in irrigated radiofrequency ablation catheter predicts lesion size and incidence of steam pop and thrombus. AB - BACKGROUND: An open-irrigated radiofrequency (RF) ablation catheter was developed to measure contact force (CF). Three optical fibers measure microdeformation of the catheter tip. The purpose of this study was to (1) validate the accuracy of CF sensor (CFS) (bench test); and (2) determine the relationship between CF and tissue temperatures, lesion size, steam pop, and thrombus during RF ablation using a canine thigh muscle preparation. METHODS AND RESULTS: CFS measurements (total 1409) from 2 catheters in 3 angles (perpendicular, parallel, and 45 degrees ) were compared with a certified balance (range, 0 to 50 g). CFS measurements correlated highly (R(2) > or =0.988; mean error, < or =1.0 g). In 10 anesthetized dogs, a skin cradle over the thigh muscle was superfused with heparinized blood at 37 degrees C. A 7F catheter with 3.5-mm saline-irrigated electrode and CFS (Endosense) was held perpendicular to the muscle at CF of 2, 10, 20, 30, and 40 g. RF was delivered (n=100) for 60 seconds at 30 or 50 W (irrigation 17 or 30 mL/min). Tissue temperature (3 and 7 mm depths), lesion size, thrombus, and steam pop increased significantly with increasing CF at each RF power. Lesion size was greater with applications of lower power (30 W) and greater CF (30 to 40 g) than at high power (50 W) with lower CF (2 to 10 g). CONCLUSIONS: This novel ablation catheter, which accurately measures CF, confirmed CF is a major determinant of RF lesion size. Steam pop and thrombus incidence also increases with CF. CFS in an open-irrigated ablation catheter that may optimize the selection of RF power and application time to maximize lesion formation and reduce the risk of steam pop and thrombus. PMID- 19808431 TI - Comparison of electroanatomic contact and noncontact mapping of ventricular scar in a postinfarct ovine model with intramural needle electrode recording and histological validation. AB - BACKGROUND: Substrate-based ablation is useful for nonhemodynamically tolerated postinfarct ventricular tachycardia. We assessed the accuracy of the CARTO contact and EnSite noncontact systems at identifying scar in a chronic ovine model with intramural plunge needle electrode recording and histological validation. METHODS AND RESULTS: Scar mapping was performed on 8 male sheep with previous percutaneous-induced myocardial infarction. Up to 20 plunge needles were inserted into the left ventricle of each animal in areas of dense scar, scar border, and normal myocardium. A simultaneous CARTO map and EnSite geometry were acquired using a single catheter, and needle electrode locations were registered. A dynamic substrate map was constructed using ratiometric 50% peak negative voltage. The scar percentage around each needle location was quantified histologically. Analysis was performed on 152 plunge needles and corresponding histological blocks. Spearman correlation with histology was 0.690 (P<0.001) for needle electrode peak-to-peak voltage (PPV), 0.362 (P<0.001) and 0.492 (P<0.001) for CARTO bipolar and unipolar PPV, and 0.381 (P<0.001) for EnSite dynamic substrate map (< or =40 mm from array). The area under the receiver operator characteristics curve (<50% and > or =50% scar) was 0.896 for needle electrode PPV, 0.726 and 0.697 for CARTO bipolar and unipolar PPV, and 0.703 for EnSite dynamic substrate map (< or=40 mm from array). CONCLUSIONS: Both the CARTO contact and EnSite noncontact systems were moderately accurate in identifying postinfarct scar when compared with intramural electrodes and confirmed with histology. The EnSite dynamic substrate map was comparable to the CARTO contact bipolar PPV when points >40 mm from the array were excluded. PMID- 19808432 TI - Malignant perinatal variant of long-QT syndrome caused by a profoundly dysfunctional cardiac sodium channel. AB - BACKGROUND: Inherited cardiac arrhythmia susceptibility contributes to sudden death during infancy and may contribute to perinatal and neonatal mortality, but the molecular basis of this risk and the relationship to genetic disorders presenting later in life is unclear. We studied the functional and pharmacological properties of a novel de novo cardiac sodium channel gene (SCN5A) mutation associated with an extremely severe perinatal presentation of long-QT syndrome in unrelated probands of different ethnicity. METHODS AND RESULTS: Two subjects exhibiting severe fetal and perinatal ventricular arrhythmias were screened for SCN5A mutations, and the functional properties of a novel missense mutation (G1631D) were determined by whole-cell patch clamp recording. In vitro electrophysiological studies revealed a profound defect in sodium channel function characterized by approximately 10-fold slowing of inactivation, increased persistent current, slowing of recovery from inactivation, and depolarized voltage dependence of activation and inactivation. Single-channel recordings demonstrated increased frequency of late openings, prolonged mean open time, and increased latency to first opening for the mutant. Subjects carrying this mutation responded clinically to the combination of mexiletine with propranolol and survived. Pharmacologically, the mutant exhibited 2-fold greater tonic and use-dependent mexiletine block than wild-type channels. The mutant also exhibited enhanced tonic (2.4-fold) and use-dependent block ( approximately 5 fold) by propranolol, and we observed additive effects of the 2 drugs on the mutant. CONCLUSIONS: Our study demonstrates the molecular basis for a malignant perinatal presentation of long-QT syndrome, illustrates novel functional and pharmacological properties of SCN5A-G1631D, which caused the disorder, and reveals therapeutic benefits of propranolol block of mutant sodium channels in this setting. PMID- 19808433 TI - Slow and discontinuous conduction conspire in Brugada syndrome: a right ventricular mapping and stimulation study. AB - BACKGROUND: Brugada syndrome (BrS) is associated with lethal arrhythmias, which are linked to specific ST-segment changes (type-1 BrS-ECG) and the right ventricle (RV). The pathophysiological basis of the arrhythmias and type-1 BrS ECG is unresolved. We studied the electrophysiological characteristics of the RV endocardium in BrS. METHODS AND RESULTS: RV endocardial electroanatomical mapping and stimulation studies were performed in controls (n=12) and BrS patients with a type-1 (BrS-1, n=10) or type-2 BrS-ECG (BrS-2, n=12) during the studies. BrS-1 patients had prominent impairment of RV endocardial impulse propagation when compared with controls, as represented by: (1) prolonged activation-duration during sinus rhythm (86+/-4 versus 65+/-3 ms), (2) increased electrogram fractionation (1.36+/-0.04 versus 1.15+/-0.01 deflections per electrogram), (3) longer electrogram duration (83+/-3 versus 63+/-2 ms), (4) activation delays on premature stimulation (longitudinal: 160+/-26 versus 86+/-9 ms; transversal: 112+/-5 versus 58+/-6 ms), and (5) abnormal transversal conduction velocity restitution (42+/-8 versus 18+/-2 ms increase in delay at shortest coupling intervals). Wider and more fractionated electrograms were also found in BrS-2 patients. Repolarization was not different between groups. CONCLUSIONS: BrS-1 and BrS-2 patients are characterized by wide and fractionated electrograms at the RV endocardium. BrS-1 patients display additional conduction slowing during sinus rhythm and premature stimulation along with abnormal transversal conduction velocity restitution. These patients may thus exhibit a substrate for slow and discontinuous conduction caused by abnormal active membrane processes and electric coupling. Our findings support the emerging notion that BrS is not solely attributable to abnormal electrophysiological properties but requires the conspiring effects of conduction slowing and tissue discontinuities. PMID- 19808434 TI - The left ventricular ostium: an anatomic concept relevant to idiopathic ventricular arrhythmias. PMID- 19808435 TI - Anomalous midline common ostium of the left and right inferior pulmonary veins: implications for pulmonary vein isolation in atrial fibrillation. PMID- 19808437 TI - Cost-effectiveness of genotyping in inherited arrhythmia syndromes: are we getting value for the money? PMID- 19808438 TI - Proof positive: efficacy of antibiotic prophylaxis in device implantation. PMID- 19808439 TI - Yield of genetic screening in inherited cardiac channelopathies: how to prioritize access to genetic testing. AB - BACKGROUND: Identification of mutations in cardiac ion channel genes concurs to the diagnosis of long-QT syndrome, Brugada syndrome, and catecholaminergic polymorphic ventricular tachycardia. However, because availability of genetic screening is still limited and reimbursement policies are lacking, there is a need of evidence-based criteria to prioritize access to genetic testing for these diseases. METHODS AND RESULTS: We determined the yield of genetic testing and cost per positive genotyping in 1394 consecutive probands. Among the 546 patients referred for long-QT syndrome-genes screening, those with clinical diagnosis of long-QT syndrome had the highest yield (64%) and lowest cost (US $8418) for each positive genotyping. Among 798 individuals screened for mutation on the SCN5A gene, the highest yield was obtained in patients with type 1 Brugada syndrome ECG pattern (51 of 405; 13%) corresponding to a cost of US $21441 per positive genotyping. In conclusive Brugada syndrome patients the presence of atrioventricular block (odds ratio: 3.3, CI: 1.8 to 6.1; P=0.0001) increases the yield (23%) of genotyping and reduces its cost (US $ 11700). Among 175 patients screened on RyR2 gene, those with documented bidirectional ventricular tachycardia had the highest incidence (62%) of mutations and the lowest cost (US $5263) per positive genotyping. Genetic screening of unselected family members of sudden cardiac death victims and idiopathic ventricular fibrillation survivors is largely ineffective (yield of 9%) and costly (US $71430 per 1 positive genotyping). CONCLUSIONS: Genotyping can be performed at reasonable cost in individuals with conclusive diagnosis of long-QT syndrome and catecholaminergic polymorphic ventricular tachycardia, and in patients with type I Brugada syndrome ECG with atrioventricular block. These patients should be given priority to access genetic testing. PMID- 19808440 TI - Absence of pathognomonic or inflammatory patterns in cardiac biopsies from patients with Brugada syndrome. AB - BACKGROUND: Brugada syndrome (BrS) is characterized by the presence of coved ST segment elevations in the right precordial leads (so-called type I ECG) and additional clinical features. Caused by cardiac ion channel gene mutations, BrS may be associated with ventricular and atrial conduction disturbances as well as ventricular fibrillation. Recent studies have discussed whether BrS is merely a primary electric disorder or whether inflammatory or other histopathologic abnormalities in the right ventricle (RV) underlie the ECG phenotype. METHODS AND RESULTS: We retrospectively analyzed BrS biopsy samples from 21 unrelated patients for histopathologic abnormalities (hypertrophy, fibrosis, inflammation, fatty tissue) together with the patients' clinical, genetic, and imaging data. Eleven patients (52%) had normal RV imaging (by angiography, echocardiography, or cardiac MRI). Results of myocardial biopsies were normal in 3 patients (14%) and revealed mostly moderate abnormalities in the others. Four patients (19%) had predominant fatty tissue in the RV myocardium. Using immunohistochemistry and conventional tissue staining, we could not detect inflammatory tissue changes, an observation compatible with the clinical absence of signs for myocarditis. CONCLUSIONS: Imaging and histopathologic evaluation may detect moderate but uncharacteristic cardiac abnormalities in patients with BrS. None of the patients had arrhythmogenic RV cardiomyopathy or overt myocarditis. Only in a small subset did predominant histopathologic abnormalities in the biopsy samples of the RV outflow tract occur that could provide a link to the ECG phenotype. A variety of mechanisms, including genetic and structural RV alterations, may underlie the Brugada ECG phenotype. PMID- 19808441 TI - Efficacy of antibiotic prophylaxis before the implantation of pacemakers and cardioverter-defibrillators: results of a large, prospective, randomized, double blinded, placebo-controlled trial. AB - BACKGROUND: Although routinely administered, definitive evidence for the benefits of prophylactic antibiotics before the implantation of permanent pacemakers and implantable cardioverter-defibrillators from a large double-blinded placebo controlled trial is lacking. The purpose of this study was to determine whether prophylactic antibiotic administration reduces the incidence of infection related to device implantation. METHODS AND RESULTS: This double blinded study included 1000 consecutive patients who presented for primary device (Pacemaker and implantable cardioverter-defibrillators) implantation or generator replacement randomized in a 1:1 fashion to prophylactic antibiotics or placebo. Intravenous administration of 1 g of cefazolin (group I) or placebo (group 2) was done immediately before the procedure. Follow-up was performed 10 days, 1, 3, and 6 months after discharge. The primary end point was any evidence of infection at the surgical incision (pulse generator pocket), or systemic infection related to be procedure. The safety committee interrupted the trial after 649 patients were enrolled due to a significant difference in favor of the antibiotic arm (group I: 2 of 314 infected patients-0.63%; group II: 11 of 335 to 3.28%; RR=0.19; P=0.016). The following risk factors were positively correlated with infection by univariate analysis: nonuse of preventive antibiotic (P=0.016); implant procedures (versus generator replacement: P=0.02); presence of postoperative hematoma (P=0.03) and procedure duration (P=0.009). Multivariable analysis identified nonuse of antibiotic (P=0.037) and postoperative hematoma (P=0.023) as independent predictors of infection. CONCLUSIONS: Antibiotic prophylaxis significantly reduces infectious complications in patients undergoing implantation of pacemakers or cardioverter-defibrillators. PMID- 19808442 TI - Left atrial posterior wall isolation does not improve the outcome of circumferential pulmonary vein ablation for atrial fibrillation: a prospective randomized study. AB - BACKGROUND: Ablation of the pulmonary veins (PVs) for atrial fibrillation treatment is often combined with linear radiofrequency lesions along the left atrium (LA) to improve the success rate. The study was designed to assess the contribution of LA posterior wall isolation to the outcome of circumferential pulmonary vein ablation (CPVA). METHODS AND RESULTS: CPVA consisted of continuous radiofrequency lesions encircling both ipsilateral PVs plus an ablation line along the mitral isthmus. Patients were then randomized into 2 groups. In the first group, superior PVs were connected by linear lesions along the LA roof (CPVA-1 group). In the second group, the LA posterior wall was isolated by adding a second line connecting the inferior aspect of the 2 inferior PVs (CPVA-2 group). The study included 120 patients (53+/-11 years, 77% male, 60% paroxysmal atrial fibrillation, LA of 41.3+/-5.4 mm, 46% with hypertension, and 22% with structural heart disease). After a single ablation procedure and a mean follow-up of 10+/-4 months, 24 (40%) patients of the CPVA-1 group had atrial fibrillation recurrences and 3 (5%) had new-onset LA flutter. In the CPVA-2 group, recurrences were due to atrial fibrillation episodes in 23 patients (38%) and LA flutter in 4 (7%). Freedom from arrhythmia recurrences was not statistically different in the CPVA-1 group as compared with the CPVA-2 group (log rank P=0.943). CONCLUSIONS: Isolation of the LA posterior wall did not increase the success rate of CPVA. PMID- 19808443 TI - Canine model of esophageal injury and atrial-esophageal fistula after applications of forward-firing high-intensity focused ultrasound and side-firing unfocused ultrasound in the left atrium and inside the pulmonary vein. AB - BACKGROUND: Left atrial-esophageal fistula is a serious and poorly understood complication of catheter ablation of atrial fibrillation. The purpose of this study was to (1) develop a canine model of esophageal injury and left atrial esophageal fistula after applications of forward-firing high-intensity focused ultrasound (HIFU) and side-firing unfocused ultrasound (SFU); (2) examine the relationship to esophageal temperature (Eso-temp); and (3) study the evolution of injury/healing. METHODS AND RESULTS: Twenty dogs were studied. After transeptal puncture, HIFU catheter (ProRhythm Inc; 13 dogs) was positioned close to the esophagus, either outside (n=6) or inside (n=7) the inferior pulmonary vein (PV). In 7 other dogs, an SFU catheter was placed deep inside the PV, close to the esophagus. A balloon (20- to 25-mm diameter) with 7 thermocouples (2-mm separation) was positioned in the esophagus (Eso-balloon). Variable air filling of the Eso-balloon controlled the distance from the esophagus to the sonication source, pressing the esophagus against left atrium/PV. One to 9 (median, 5) HIFU (35 W) and 5 to 7 (median, 5) SFU (40 W) sonications were delivered for 40 seconds. Maximum luminal Eso-temp was closely related to HIFU Eso-balloon distance. For HIFU outside PV, Eso-temp >or=50 degrees C occurred only for HIFU Eso-balloon distance or=50 degrees C, with HIFU Eso-balloon distance up to 6.8 mm. Endoscopy identified esophageal ulcer immediately after ablation in 11 of 13 HIFU dogs and 7 of 7 SFU dogs, all with Eso-temp >or=50 degrees C. Endoscopy at 2 weeks showed ulcer healing in 5 of 11 chronic dogs and ulcer size progression with relaxation of the lower esophageal sphincter and esophagitis in 6 dogs. Two dogs developed left atrial-esophageal fistula and died at 2 weeks. CONCLUSIONS: This model produces esophageal ulcer when Eso-temp is >or=50 degrees C. Eso-temp is higher with HIFU/SFU applications closer to the esophagus and with HIFU/SFU applications inside the PV. Ulcer progression and left atrial-esophageal fistula were associated with reflux esophagitis. PMID- 19808444 TI - Applications of cardiac magnetic resonance in electrophysiology. PMID- 19808445 TI - P wave indices: current status and future directions in epidemiology, clinical, and research applications. PMID- 19808446 TI - Is there a significant transmural gradient in repolarization time in the intact heart? Cellular basis of the T wave: a century of controversy. PMID- 19808447 TI - Is there a significant transmural gradient in repolarization time in the intact heart?: Repolarization Gradients in the Intact Heart. PMID- 19808448 TI - An "almost wide" QRS tachycardia. PMID- 19808449 TI - Fibrillation in the superior vena cava mimicking atrial tachycardia. PMID- 19808450 TI - Cardiac arrest related to coronary vasospasm in a patient with long QT1. PMID- 19808451 TI - WPW pattern in the asymptomatic individual: has anything changed? PMID- 19808452 TI - The modulated dispersion hypothesis confirmed in humans. PMID- 19808453 TI - Asymptomatic ventricular preexcitation: a long-term prospective follow-up study of 293 adult patients. AB - BACKGROUND: Sudden cardiac death can be the first clinical presentation of asymptomatic ventricular preexcitation. METHODS AND RESULTS: From 1995 to 2005, we prospectively collected clinical and electrophysiological data among 293 adults with asymptomatic ventricular preexcitation (61.4% males; median age, 36 years; interquartile range [IQR], 28 to 47.5). After electrophysiological testing, patients were prospectively followed, taking no drugs. The primary end point of the study was the occurrence of a first arrhythmic event. Predictors of arrhythmic events were analyzed by univariate and multivariate Cox models. Over a median follow-up of 67 months (minimum to maximum, 8 to 90), after electrophysiological testing, 262 patients (median age, 37 years; IQR, 30 to 48) did not experience arrhythmic events, remaining totally asymptomatic, whereas 31 patients (median age, 25 years; IQR, 22 to 29; median follow-up, 27 months; minimum to maximum, 8 to 55) had a first arrhythmic event, which was potentially life-threatening in 17 of them (median age, 24 years; IQR, 20 to 28.5; median follow-up, 25 months; minimum to maximum, 9 to 55). Potentially life-threatening tachyarrhythmias resulted in resuscitated cardiac arrest (1 patient), presyncope (7 patients) syncope (4 patients), or dizziness (5 patients). In multivariate analysis age (P=0.004), inducibility (P=0.001) and anterograde effective refractory period of the accessory pathway < or = 250 ms (P=0.001) predicted potentially life-threatening arrhythmias. CONCLUSIONS: These results indicate that prognosis of adults who present with asymptomatic ventricular preexcitation is good, and the risk of a significant event is small. Short anterograde effective refractory period of the accessory pathway and inducibility at baseline are independent predictors of potentially life-threatening arrhythmic events, and the risk decreases with increasing age. PMID- 19808454 TI - Esophageal capsule endoscopy after radiofrequency catheter ablation for atrial fibrillation: documented higher risk of luminal esophageal damage with general anesthesia as compared with conscious sedation. AB - BACKGROUND: Left atrioesophageal fistula is a rare but devastating complication that may occur after catheter ablation of atrial fibrillation. We used capsule endoscopy to assess esophageal injury after catheter ablation for atrial fibrillation in a population randomized to undergo general anesthesia or conscious sedation. METHODS AND RESULTS: Fifty patients undergoing atrial fibrillation ablation for paroxysmal symptomatic atrial fibrillation refractory to antiarrhythmic drugs were enrolled and randomized, including those undergoing the procedure under general anesthesia (25 patients, group 1) and those receiving conscious sedation with fentanyl or midazolam (25 patients, group 2). All patients underwent esophageal temperature monitoring during the procedure. The day after ablation, all patients had capsule endoscopy to assess the presence of endoluminal tissue damage of the esophagus. We observed esophageal tissue damage in 12 (48%) patients of group 1 and 1 esophageal tissue damage in a single patient (4%) of group 2 (P<0.001). The maximal esophageal temperature was significantly higher in patients undergoing general anesthesia (group 1) versus patients undergoing conscious sedation (group 2) (40.6+/-1 degrees C versus 39.6+/-0.8 degrees C; P< 0.003). The time to peak temperature was 9+/-7 seconds in group 1 and 21+/-9 seconds in group 2, and this difference was statistically significant (P<0.001). No complication occurred during or after the administration of the pill cam or during the procedures. All esophageal lesions normalized at the 2-month repeat endoscopic examination. CONCLUSIONS: The use of general anesthesia increases the risk of esophageal damage detected by capsule endoscopy. PMID- 19808455 TI - Atrial fibrillation ablation strategies for paroxysmal patients: randomized comparison between different techniques. AB - BACKGROUND: Whether different ablation strategies affect paroxysmal atrial fibrillation (AF) long-term freedom from AF/atrial tachyarrhythmia is unclear. We sought to compare the effect of 3 different ablation approaches on the long-term success in patients with paroxysmal AF. METHODS AND RESULTS: One hundred three consecutive patients with paroxysmal AF scheduled for ablation and presenting in the electrophysiology laboratory in AF were selected for this study. Patients were randomized to pulmonary vein antrum isolation (PVAI; n=35) versus biatrial ablation of the complex fractionated atrial electrograms (CFAEs; n=34) versus PVAI followed by CFAEs (n=34). Patients were given event recorders and followed up at 3, 6, 9, 12, and 15 months postablation. There was no statistical significant difference between the groups in term of sex, age, AF duration, left atrial size, and ejection fraction. At 1 year follow-up, freedom from AF/atrial tachyarrhythmia was documented in 89% of patients in the PVAI group, 91% in the PVAI plus CFAEs group, and 23% in the CFAEs group (P<0.001) after a single procedure and with antiarrhythmic drugs. CONCLUSIONS: No difference in terms of success rate was seen between PVAI alone and PVAI associated with defragmentation. CFAEs ablation alone had the smallest impact on AF recurrences at 1-year follow-up. These results suggest that antral isolation is sufficient to treat most patients with paroxysmal AF. PMID- 19808456 TI - Remote robotic navigation and electroanatomical mapping for ablation of atrial fibrillation: considerations for navigation and impact on procedural outcome. AB - BACKGROUND: Radiofrequency current ablation of atrial fibrillation (AF) requires high technical skills to achieve optimal catheter stability and is associated with an individually high x-ray exposure to both the patient and the operator. To facilitate catheter navigation and to reduce the operator's x-ray burden, remote navigation (RN) systems have been developed. Considerations for navigation of a novel remote robotic navigation system in pulmonary vein isolation (PVI) procedures are reported. METHODS AND RESULTS: In 65 patients with drug-refractory AF (43 paroxysmal, 22 persistent), complete circumferential PVI was performed using RN in conjunction with different electroanatomic mapping systems. Acute complete PVI using exclusively RN was achieved in 95%. The procedure time was 195+/-40 minutes. The operator's x-ray exposure time was reduced by 6+/-4 minutes (35%) using RN. In 7 of 14 patients with persistent AF, conversion to sinus rhythm was achieved by radiofrequency current ablation. During a median follow-up period of 239 days (range, 184 to 314 days), 47 of 65 patients (73%) remained free of any documented atrial tachyarrhythmia recurrences after a single procedure. The relative proportion of patients remaining free of AF was 76% and 68% for paroxysmal and persistent AF, respectively. CONCLUSIONS: PVI using the novel RN system can be performed safely and effectively. One third of the operator's fluoroscopy exposure time might be saved using RN. However, the questions of whether the overall fluoroscopy exposure is reduced by RN and whether RN improves PVI procedures needs to be assessed during a comparative trial between man and machine. PMID- 19808457 TI - Risk factors for mortality in patients with cardiac device-related infection. AB - BACKGROUND: Because of the increased use of pacemakers and implantable cardioverter defibrillators, infection has become a complication with significant morbidity and mortality. Data on risk factors for mortality in patients with cardiac-device related infection are limited. We evaluated the prognostic significance of key clinical and echocardiographic variables in a large retrospective population of patients with cardiac-device related infection. METHODS AND RESULTS: Two hundred ten patients with cardiac-device related infection were identified at the University of Michigan between 1995 and 2006. Data were abstracted on key clinical and echocardiographic variables, treatment strategy, and 6-month outcomes. We used multivariable Cox proportional hazards models to examine clinical and echocardiographic variables that were associated with 6-month mortality. Mean age for our study population was 63+/-17 years, and 72 (44%) were women. All-cause 6-month mortality was 18% (n=37). Independent variables associated with death were systemic embolization (hazard ratio 7.11; 95% CI 2.74 to 18.48), moderate or severe tricuspid regurgitation (hazard ratio 4.24; 95% CI 1.84 to 9.75), abnormal right ventricular function (hazard ratio 3.59; 95% CI 1.57 to 8.24), and abnormal renal function (hazard ratio 2.98; 95% CI 1.17 to 7.59). Size and mobility of cardiac device vegetations were not independently associated with mortality. CONCLUSIONS: We identified several clinical and echocardiographic variables that identify patients with cardiac device related infection who are at high-risk for mortality and may benefit from more aggressive evaluation. PMID- 19808458 TI - Acute effects of right ventricular apical pacing on left ventricular synchrony and mechanics. AB - BACKGROUND: Chronic right ventricular (RV) apical pacing has a detrimental effect on left ventricular (LV) function. However, the acute effects of RV apical pacing on LV mechanics remain unclear. The purpose of the study was to assess the acute impact of RV apical pacing on global LV function, evaluating LV contraction synchrony and LV shortening and twist, using 2D speckle-tracking strain imaging. METHODS AND RESULTS: A group of 25 patients with structural normal hearts referred for electrophysiological study were studied. Two-dimensional echocardiography was performed at baseline and during RV apical pacing at the time of the electrophysiological study. Changes in LV synchrony and mechanics (longitudinal shortening and twist) were assessed using speckle-tracking strain imaging. In addition, 25 controls matched by age, sex, and LV function were studied during sinus rhythm. The group of patients (44+/-12 years, 10 men) and the group of controls (48+/-3 years, 8 men) showed comparable LV synchrony, LV longitudinal shortening, and LV twist at baseline. However, during RV apical pacing, a more dyssynchronous LV contraction was observed in the patients (from 21 ms [Q(1):10, Q(3):53] to 91 ms [Q(1):40, Q(3):204], P<0.001) together with an impairment in LV longitudinal shortening (from -18.3+/-3.5% to -11.8+/-3.6%, P<0.001) and in LV twist (from 12.4+/-3.7 degrees to 9.7+/-2.6 degrees , P=0.001). CONCLUSIONS: During RV apical pacing, an acute induction of LV dyssynchrony is observed. In addition, LV longitudinal shortening and LV twist are acutely impaired. PMID- 19808459 TI - A randomized study to compare ramp versus burst antitachycardia pacing therapies to treat fast ventricular tachyarrhythmias in patients with implantable cardioverter defibrillators: the PITAGORA ICD trial. AB - BACKGROUND: In patients with implantable cardioverter-defibrillators (ICDs), antitachycardia pacing (ATP) is highly effective in terminating fast ventricular tachycardias (FVTs) and lowers the use of high-energy shocks, without increasing the risk of arrhythmia acceleration or syncope. METHODS AND RESULTS: The aim of the PITAGORA ICD trial was to randomly compare 2 ATP strategies (88% coupling interval burst versus 91% coupling interval ramp, both 8 pulses) in terms of ATP efficacy, arrhythmia acceleration, and syncope. Two hundred six ICD patients (83% male, 67+/-11 years) were enrolled. FVT episodes with cycle lengths between 240 and 320 ms were treated by 1 ATP sequence and, in the event of failure, by shocks. Over a median follow-up of 36 months, 829 spontaneous ventricular tachyarrhythmia episodes were detected in 79 patients. Episode review identified 595 episodes as true ventricular arrhythmias in 72 patients; devices classified 111 (18.7%) episodes as VF, 216 (36.3%) as FVT, and 268 (45.0%) as VT. Fifty-six patients had 214 treated FVT episodes-2 FVTs self-terminated before ATP release; 44 (79%) of these had at least 1 effective ATP intervention, and 34 (61%) were spared ICD shocks. Burst terminated 100 of 133 (75.2%) FVT episodes, whereas ramp terminated 44 of 81 (54.3%; P=0.015). Acceleration occurred in 9 of 214 (4.2%) FVT episodes treated: 6 episodes in 3 ramp patients and 3 episodes in 3 burst patients. Two patients-1 in each group-suffered 1 syncopal event associated to a nonterminated FVT episode. CONCLUSIONS: Burst is significantly more efficacious than ramp in terminating FVT episodes. As the first therapy for FVT episodes, ATP carries a low risk of acceleration or syncopal events. PMID- 19808460 TI - Inferior and lateral electrocardiographic repolarization abnormalities in Brugada syndrome. AB - BACKGROUND: Repolarization abnormalities in the inferior-lateral leads in Brugada syndrome (BS) have not been systematically investigated. METHODS AND RESULTS: 280 patients (age, 41+/-18 years; 168 males) with BS were screened for inferior lateral repolarization abnormalities. The repolarization abnormalities were classified either as early repolarization pattern or coved > or = 2-mm Brugada pattern and as spontaneous or class I antiarrhythmic drug (AAD) induced. Thirty two patients (11%) had inferior-lateral spontaneous early repolarization pattern. These patients were less likely to be asymptomatic at first presentation (13 of 32 versus 156 of 248 patients, P=0.02), and spontaneous type I ECG was more frequent among them (38% versus 21%, P=0.05). The spontaneous early repolarization pattern occurred more frequently among patients with BS than in 283 family members not having BS (11% versus 6%, P=0.03). Class I AAD administration provoked inferior-lateral coved Brugada pattern in 13 patients with BS. These patients had longer baseline PR intervals (206+/-48 versus 172+/ 31 ms, P<0.001) and class I AAD-induced QRS interval prolongation (108 to 178 versus 102 ms to 131 ms, P<0.001). In 3 patients, the class I AAD-provoked coved Brugada pattern was only present in the inferior leads. CONCLUSIONS: Inferior lateral early repolarization pattern occurs spontaneously relatively frequently in BS. These patients have a more severe phenotype. Class I AAD administration provokes inferior-lateral coved Brugada pattern in 4.6% of patients. We report for the first time 3 patients in whom the class I AAD-provoked coved Brugada pattern was only observed in the inferior leads. PMID- 19808461 TI - Interaction of activation-repolarization coupling and restitution properties in humans. AB - BACKGROUND: Dynamic modulation of repolarization is important in arrhythmogenesis. An inverse relation exists in myocardium between activation time (AT) and action potential duration (APD). We hypothesized that resulting gradients of APD and diastolic interval (DI) interact with restitution properties and modulate the timing of repolarization. METHODS AND RESULTS: Activation recovery intervals (ARI) were acquired from reconstructed noncontact unipolar electrograms from the left ventricular endocardium in 9 patients (7 male) with normal ventricles. At a basic paced cycle length (median, 450 ms), ARIs shortened along the path of activation, with a mandatory reciprocal increase of DIs. In the median patient, this range of DIs started at 230 ms at the site of earliest activation and increased to 279 ms at the site of latest activation at a basic cycle length of 450 ms. Four consecutive standard S1 to S2 restitution curves were performed. At sites with a longer ARI (and therefore shorter DI) close to the site of stimulation, premature stimulation produced more shortening of ARIs; therefore, the time course of restitution was steeper than at more distal sites. At normal heart rate, the decrease in ARIs along the conduction pathway compensated for later activation. Thus, dispersion in repolarization time (RT) is smaller than dispersion in ARI in a heart with a steep negative AT-ARI relationship. This protective effect is lost in hearts without such a relationship. In the patients with a steep AT-ARI relationship at basic cycle length, this relation is lost after premature stimulation and is a function of prematurity. Thus, dispersion in RT is larger after shortly coupled extra stimuli in patients with a steep AT-ARI relationship. CONCLUSIONS: A complex interplay exists between activation-repolarization coupling and restitution properties, largely driven by ARI and DI gradients. This plays a significant role in the dynamics of repolarization in humans. PMID- 19808462 TI - Effects of chronic gap junction conduction-enhancing antiarrhythmic peptide GAP 134 administration on experimental atrial fibrillation in dogs. AB - BACKGROUND: Abnormal intercellular communication caused by connexin dysfunction may contribute to atrial fibrillation (AF). The present study assessed the effect of the gap junction conduction-enhancing antiarrhythmic peptide GAP-134 on AF inducibility and maintenance in a dog model of atrial cardiomyopathy. METHODS AND RESULTS: Twenty-four dogs subject to simultaneous atrioventricular pacing (220 bpm for 14 days) were randomly assigned to placebo treatment (PACED-CTRL; 12 dogs) or oral GAP-134 (2.9 mg/kg BID; PACED-GAP-134; 12 dogs) starting on day 0. UNPACED-CTRL (4 dogs) and UNPACED-GAP-134 (4 dogs) served as additional control groups. Change in left atrial (LA) systolic area from baseline to 14 days was calculated using transoesophageal echocardiography. At 14 days, animals underwent an open-chest electrophysiological study. PACED-CTRL dogs (versus UNPACED-CTRL) had a shorter estimated LA wavelength (8.0+/-1.4 versus 24.4+/-2.5 cm, P<0.05) and a greater AF vulnerability (mean AF duration, 1588+/-329 versus 25+/-34 seconds, P<0.05). Oral GAP-134 had no effect on AF vulnerability in UNPACED dogs. Compared with PACED-CTRL dogs, PACED-GAP-134 dogs had a longer estimated LA wavelength (10.2+/-2.8 versus 8.0+/-1.4 cm, respectively, P<0.05). Oral GAP-134 did not significantly reduce AF inducibility or maintenance in the entire group of 24 PACED dogs; in a subgroup of dogs (n=11) with less than 100% increase in LA systolic area, oral GAP-134 reduced AF induction from 100% to 40% and mean AF duration from 1737+/-120 to 615+/-280 seconds (P<0.05). CONCLUSIONS: Oral GAP-134 reduces pacing-induced decrease in LA wavelength and appears to attenuate AF vulnerability in dogs with less atrial mechanical remodeling. Gap junction modulation may affect AF in some circumstances. PMID- 19808463 TI - Preshock cardiopulmonary resuscitation worsens outcome from circulatory phase ventricular fibrillation with acute coronary artery obstruction in swine. AB - BACKGROUND: Some clinical studies have suggested that chest compressions before defibrillation improve survival in cardiac arrest because of prolonged ventricular fibrillation (VF; ie, within the circulatory phase). Animal data have also supported this conclusion, and we have previously demonstrated that preshock chest compressions increase the VF median frequency and improve the likelihood of a return of spontaneous circulation in normal swine. We hypothesized that chest compressions before defibrillation in a swine model of acute myocardial ischemia would also increase VF median frequency and improve resuscitation outcome. METHODS AND RESULTS: Twenty-six swine were subjected to balloon occlusion of the left anterior descending coronary artery for 2 hours. The balloon was removed and VF was induced and untreated for 8 minutes. Swine were then treated with up to 3 stacked defibrillation shocks (n=13, shock-first group) or 3 minutes of chest compressions before shock (n=13, preshock cardiopulmonary resuscitation group). In the preshock cardiopulmonary resuscitation group, median frequency was increased from 7.0+/-0.8 to 13.9+/-1.6 Hz after chest compressions (P=0.002). Despite the improved median frequency in the preshock cardiopulmonary resuscitation group, 24-hour survival with favorable neurological status was significantly worse in the preshock cardiopulmonary resuscitation group (1/13) compared with the shock-first group (8/13, P=0.01). CONCLUSIONS: In a swine model of prolonged VF in acute myocardial ischemia, 24-hour survival with favorable neurological status was more likely when defibrillation was performed first without preceding chest compressions. Myocardial substrate is an important factor in determining the optimal resuscitation strategy. PMID- 19808464 TI - Cardiac ion channels. PMID- 19808465 TI - Development of the cardiac conduction system: why are some regions of the heart more arrhythmogenic than others? PMID- 19808466 TI - Noninvasive evaluation of radiofrequency lesions in the human ventricular myocardium by contrast-enhanced cardiac magnetic resonance. PMID- 19808467 TI - Letter by Khan et al regarding article "Efficacy of antibiotic prophylaxis before the implantation of pacemakers and cardioverter-defibrillators: results of a large, prospective, randomized, double-blinded, placebo-controlled trial". PMID- 19808469 TI - Atrial fibrillation through the patient's eyes. PMID- 19808470 TI - Symptoms in atrial fibrillation: why keep score? PMID- 19808471 TI - Validation of a new simple scale to measure symptoms in atrial fibrillation: the Canadian Cardiovascular Society Severity in Atrial Fibrillation scale. AB - BACKGROUND: Atrial fibrillation (AF) is commonly associated with impaired quality of life. There is no simple validated scale to quantify the functional illness burden of AF. The Canadian Cardiovascular Society Severity in Atrial Fibrillation (CCS-SAF) scale is a bedside scale that ranges from class 0 to 4, from no effect on functional quality of life to a severe effect on life quality. This study was performed to validate the scale. METHODS AND RESULTS: In 484 patients with documented AF (62.2+/-12.5 years of age, 67% men; 62% paroxysmal and 38% persistent/permanent), the SAF class was assessed and 2 validated quality-of-life questionnaires were administered: the SF-36 generic scale and the disease specific AFSS (University of Toronto Atrial Fibrillation Severity Scale). There is a significant linear graded correlation between the SAF class and measures of symptom severity, physical and emotional components of quality of life, general well-being, and health care consumption related to AF. Patients with SAF class 0 had age- and sex-standardized SF-36 scores of 0.15+/-0.16 and -0.04+/-0.31 (SD units), that is, units away from the mean population score for the mental and physical summary scores, respectively. For each unit increase in SAF class, there is a 0.36 and 0.40 SD unit decrease in the SF-36 score for the physical and mental components. As the SAF class increases from 0 to 4, the symptom severity score (range, 0 to 35) increases from 4.2+/-5.0 to 18.4+/-7.8 (P<0.0001). CONCLUSIONS: The CCS-SAF scale is a simple semiquantitative scale that closely approximates patient-reported subjective measures of quality of life in AF and may be practical for clinical use. PMID- 19808472 TI - Feasibility of the radiofrequency hot balloon catheter for isolation of the posterior left atrium and pulmonary veins for the treatment of atrial fibrillation. AB - BACKGROUND: Atrial fibrillation originates mostly from the pulmonary vein (PV) foci or non-PV foci in the posterior left atrium (LA). The present study was designed to evaluate the feasibility and safety of a novel radiofrequency hot balloon catheter for the treatment of patients with atrial fibrillation by electrically isolating the posterior LA, including all PVs. METHODS AND RESULTS: One hundred consecutive patients with drug-resistant atrial fibrillation (63 paroxysmal, 37 persistent) were enrolled. The isolation of the PVs was performed by wedging the balloon at each PV antrum to create circumferential lesions in each case. Contiguous linear lesions were also created at the roof between the superior PVs and at the bottom of the posterior LA between the inferior PVs by dragging the balloon along the endocardium. Complete elimination of the posterior LA and PV potentials was achieved in all 100 cases, confirmed by either conventional or electro-anatomic mapping system. The total procedure time was 129+/-26 minutes, inclusive of 29.9+/-7.3 minutes of fluoroscopy time. Follow-up during 11.0+/-4.8 months confirmed that 92 patients (60 paroxysmal, 32 persistent) were free from atrial fibrillation without antiarrhythmic drugs, and in the remaining patients except for 2 with LA tachycardia, sinus rhythm was maintained with antiarrhythmic drugs. With precautions of esophageal cooling by irrigation dictated by temperature monitoring and monitoring phrenic nerve pacing, no LA-esophageal fistula or permanent phrenic nerve injury occurred. CONCLUSIONS: This feasibility study supports the safety and efficacy of radiofrequency hot balloon catheter for complete isolation of the posterior LA and PVs. PMID- 19808473 TI - Spatiotemporal organization of the left atrial substrate after circumferential pulmonary vein isolation of atrial fibrillation. AB - BACKGROUND: There is a paucity of data regarding the mechanism of maintaining atrial fibrillation (AF) after pulmonary vein isolation (PVI) in patients with AF. The aim of this study was to examine the impact of circumferential PVI on the left atrial (LA) substrate characteristics. METHODS AND RESULTS: Seventy-two AF patients (age, 53+/-11 years) underwent mapping and catheter ablation using an NavX system. The biatrial characteristics such as the complex fractionated atrial electrograms (CFEs; based on fractionated intervals) and frequency analysis (based on dominant frequencies) were mapped before and after PVI. PVI with electric isolation was performed in all patients. In the 45 patients who did not respond to PVI, the continuous CFEs (>8 seconds, 18+/-18% and 12+/-17% of the LA sites, before and after PVI, respectively, P=0.02), degree of LA fractionation (mean fractionated interval: 75.6+/-14.3 msec versus 87.3+/-16.7 msec, P=0.001), and mean LA dominant frequencies (6.92+/-0.88 Hz versus 6.58+/-0.91 Hz, P=0.001) decreased after PVI. Complete PVI altered the distribution of the CFEs toward the LA anteroseptum, mitral annulus, and LA appendage regions. A persistent presence of continuous CFEs in the vicinity of the dominant frequencies sites (observed in 53% patients) correlated with a higher procedural AF termination rate for the CFE ablation (63% versus 23%, P<0.05). CONCLUSIONS: Complete PVI eliminated some CFEs in the LA and altered the distribution of the CFEs. The persistent presence of CFEs before and after PVI in the vicinity of the high frequency sites is important for AF maintenance after PVI. PMID- 19808474 TI - Long-term outcome of ablative therapy of postoperative supraventricular tachycardias in patients with univentricular heart: a European multicenter study. AB - BACKGROUND: Catheter ablation has evolved as a possible curative treatment modality for supraventricular tachycardias (SVT) in patients with univentricular heart. However, the long-term outcome of ablation procedures is unknown. We evaluated the procedural and long-term outcome of ablative therapy of late postoperative SVT in patients with univentricular heart. METHODS AND RESULTS: Patients with univentricular heart (n=19, 11 male; age, 29+/-9 years) referred for ablation of SVT were studied. Ablation was guided by 3D electroanatomic mapping in all but 2 procedures. A total of 41 SVT were diagnosed as intra-atrial reentrant tachycardia (n=30; cycle length, 310+/-68 ms), typical atrial flutter (n=4; cycle length, 288+/-42 ms), focal atrial tachycardia (n=6; cycle length, 400+/-60 ms), and atrial fibrillation (n=1). Ablation was successful in 73% of intra-atrial reentrant tachycardia, 75% of atrial flutter, and all focal atrial tachycardia and focal atrial fibrillation. During the follow-up period of 53+/-34 months, 2 patients were lost to follow-up, 3 died of heart failure, 2 underwent heart transplantation, and 1 underwent conduit replacement. Of the remaining group, 8 had sinus rhythm and 3 had SVT. CONCLUSIONS: Focal and reentrant mechanisms underlie postoperative SVT in patients with univentricular heart. Successive SVT developing over time may be caused by different mechanisms. Ablative therapy is potentially curative, with a procedural success rate of 78%. In patients who had multiple ablation procedures, the SVT originated from different atrial sites, suggesting that these new SVT were caused by progressive atrial disease. Despite recurrent SVT, sinus rhythm at the end of the follow-up period was achieved in 72%. PMID- 19808475 TI - Long-term improvement in left ventricular strain after successful catheter ablation for atrial fibrillation in patients with preserved left ventricular systolic function. AB - BACKGROUND: The effect of successful catheter ablation on left ventricular (LV) strain in patients with preserved LV systolic function is unknown. The aim of the present study was to assess the long-term effects of catheter ablation for atrial fibrillation (AF) on LV strain and strain rate in patients with preserved LV ejection fraction. METHODS AND RESULTS: In 78 patients undergoing catheter ablation for AF, speckle tracking strain imaging was performed to assess LV strain in 3 directions (radial, circumferential, and longitudinal) at baseline and after 12-month follow-up. The study population was divided into 2 groups, according to the maintenance of sinus rhythm (SR) during follow-up. After 13.8+/ 4.7 months of follow-up, 54 patients (69%) were in SR (SR group), whereas 24 patients (31%) had recurrence of AF (AF group). No significant changes in LV ejection fraction from baseline to follow-up were noted (60+/-7% versus 59+/-7%, P=NS). Circumferential strain improved significantly in the SR group (-18.3+/ 3.2% versus -20.4+/-3.8%, P<0.001), whereas it remained unchanged in the AF group (-18.9+/-3.5% versus -17.9+/-3.1%, P=NS). In the SR group, significant improvements in LV longitudinal strain and strain rate were noted, whereas in the AF group, LV longitudinal strain and strain rate deteriorated significantly at long-term follow-up. CONCLUSIONS: After successful catheter ablation, LV circumferential and longitudinal strain and strain rate improve significantly in patients who maintain SR. In contrast, a decrease in LV longitudinal strain and strain rate is observed in patients with recurrence of AF. PMID- 19808476 TI - Feasibility of real-time MRI with a novel carbon catheter for interventional electrophysiology. AB - BACKGROUND: Cardiac MRI offers 3D real-time imaging with unsurpassed soft tissue contrast without x-ray exposure. To minimize safety concerns and imaging artifacts in MR-guided interventional electrophysiology (EP), we aimed at developing a setup including catheters for ablation therapy based on carbon technology. METHODS AND RESULTS: The setup, including a steerable carbon catheter, was tested for safety, image distortion, and feasibility of diagnostic EP studies and radiofrequency ablation at 1.5 T. MRI was performed in 3 different 1.5-T whole-body scanners using various receive coils and pulse sequences. To assess unintentional heating of the catheters by radiofrequency pulses of the MR scanner in vitro, a fluoroptic thermometry system was used to record heating at the catheter tip. Programmed stimulation and ablation therapy was performed in 8 pigs. There was no significant heating of the carbon catheters while using short, repetitive radiofrequency pulses from the MR system. Because there was no image distortion when using the carbon catheters, exact targeting of the lesion sites was possible. Both atrial and ventricular radiofrequency ablation procedures including atrioventricular node modulation were performed successfully in the scanner. Potential complications such as pericardial effusion after intentional perforation of the right ventricular free wall during ablation could be monitored in real time as well. CONCLUSIONS: We describe a newly developed EP technology for interventional electrophysiology based on carbon catheters. The feasibility of this approach was demonstrated by safety testing and performing EP studies and ablation therapy with carbon catheters in the MRI environment. PMID- 19808477 TI - Mutations in sodium channel beta1- and beta2-subunits associated with atrial fibrillation. AB - BACKGROUND: We and others have reported mutations in the cardiac predominant sodium channel gene SCN5A in patients with atrial fibrillation (AF). We also have reported that SCN1B is associated with Brugada syndrome and isolated cardiac conduction disease. We tested the hypothesis that mutations in the 4 sodium channel beta-subunit genes SCN1B-SCN4B contribute to AF susceptibility. METHODS AND RESULTS: Screening for mutations in the 4 beta-subunit genes was performed in 480 patients with AF (118 patients with lone AF and 362 patients with AF and cardiovascular disease) and 548 control subjects (188 ethnically defined anonymized subjects and 360 subjects without AF). The effects of mutant beta subunits on SCN5A mediated currents were studied using electrophysiological studies. We identified 2 nonsynonymous variants in SCN1B (resulting in R85H, D153N) and 2 in SCN2B (R28Q, R28W) in patients with AF. These occur at residues highly conserved across mammals and were absent in control subjects. In 3 of 4 mutation carriers, the ECGs showed saddleback-type ST-segment elevation in the right precordial leads. Transcripts encoding both SCN1B and SCN2B were detected in human atrium and ventricle. In heterologous expression studies using Chinese hamster ovary cells, the mutant beta1- or beta2-subunits reduced SCN5A-mediated current and altered channel gating compared with coexpression of wild-type subunits. CONCLUSIONS: Loss of function mutations in sodium channel beta-subunits were identified in patients with AF and were associated with a distinctive ECG phenotype. These findings further support the hypothesis that decreased sodium current enhances AF susceptibility. PMID- 19808478 TI - Prediction of ventricular tachyarrhythmias by intracardiac repolarization variability analysis. AB - BACKGROUND: Arrhythmic sudden cardiac death (SCD) is generally mediated by ventricular fibrillation (VF) or fast ventricular tachycardia (FVT). We studied the predictive value of temporal QT variability detected from various sources of cardiac electric signal: surface ECG, far-field (FF), and near-field (NF) intracardiac electrograms (EGMs) in patients with implantable cardioverter defibrillators (ICDs). METHODS AND RESULTS: Surface ECG and FF and NF intracardiac EGMs were simultaneously recorded at rest (mean heart rate, 74+/-15 bpm) for 4.5+/-1.3 minutes in 298 patients (mean age, 59+/-14; 216 male [73%]) with structural heart disease and an implanted Medtronic ICD for primary (231 patients, 78%) or secondary (67 patients, 22%) prevention of SCD. During mean follow-up of 16+/-8 months, 52 (13.1% per person-year of follow-up) patients sustained VT/VF and received appropriate ICD therapies, but only 19 (4.8% per person-year of follow-up) patients sustained FVT/VF with cycle length 30 seconds' duration occurring >90 days after surgery. Mean follow-up was 516+/-181 days (202 to 858 days). Twenty-eight (65%) patients had no atrial tachyarrhythmia >30 seconds by 1 year, and 15 (35%) patients had atrial tachyarrhythmia recurrences by 1 year. Eight of 15 patients with recurrent AF had catheter ablation resulting in elimination and/or reduction of AF episodes in 7 of 8 patients. Four of 15 patients had AF elimination or reduction with antiarrhythmic drugs alone. Three patients did not benefit from surgery and received rate control only. There were no deaths; 1 phrenic nerve injury and 2 pleural effusions were the only major complications. CONCLUSIONS: The single procedure success at 1-year follow-up for surgical pulmonary vein isolation and ganglionic plexi ablation is 65%. Atrial tachyarrhythmia recurrences after surgery are usually responsive to catheter ablation and/or antiarrhythmic drugs. PMID- 19808493 TI - Familial aggregation of atrial fibrillation: a study in Danish twins. AB - BACKGROUND: Heritability may play a role in nonfamilial atrial fibrillation (AF). We hypothesized that a monozygotic (MZ) twin whose co-twin was diagnosed with AF would have an increased risk of the disease compared with a dizygotic (DZ) twin in the same situation. METHODS AND RESULTS: A sample of 1137 same-sex twin pairs (356 MZ and 781 DZ pairs) in which one or both members were diagnosed with AF were identified in The Danish Twin Registry. Concordance rates were twice as high for MZ pairs than for DZ pairs regardless of sex (22.0% versus 11.6%, P<0.0001). In a Cox regression of event-free survival times, we compared the time span between occurrences of disease in MZ and DZ twins. The unaffected twin was included when his or her twin-sibling (the index twin) was diagnosed with AF. After adjustment for age at entry, MZ twins had a significantly shorter event free survival time (hazard ratio, 2.0; 95% CI, 1.3 to 3.0), thereby indicating a genetic component. Using biometric models, we estimated the heritability of AF to be 62% (55% to 68%), due to additive genetics. There were no significant differences across sexes. CONCLUSIONS: All the analyses of twin similarities in the present study indicate that genetic factors play a substantial role in the risk of AF for both sexes. The recurrence risk for co-twins (12% to 22%) is clinically relevant and suggests that co-twins of AF-affected twins belong to a high-risk group for AF. PMID- 19808494 TI - An acute experimental model demonstrating 2 different forms of sustained atrial tachyarrhythmias. AB - BACKGROUND: The objective of this study was to develop an acute experimental model showing both focal and macroreentrant sustained atrial fibrillation (AF). METHODS AND RESULTS: In 31 anesthetized dogs, bilateral thoracotomies allowed the attachment of electrode catheters at the right and left superior pulmonary veins, atrial free walls, and atrial appendages. Acetylcholine, 100 mmol/L, was applied topically to either appendage. Sequential radiofrequency ablation was achieved for the ganglionated plexi (GP), found adjacent to the 4 pulmonary veins. In 12 separate studies, a propafenone bolus, 2 mg/kg, was given before and after GP ablations at the start of acetylcholine-induced AF. Acetylcholine caused abrupt onset of AF (n=22) or induced AF by burst pacing (n=9) that lasted > or = 10 minutes. Rapid, regular, or fractionated atrial electrograms were consistently seen (average cycle length, 37+/-7 ms) at the appendages versus cycle lengths of 114+/-23 ms at other atrial sites. After ablations of GP, AF abruptly terminated (n=25). In 6 dogs, sustained atrial tachyarrhythmias continued. Pacing at specific atrial sites organized electrograms of one atrium or also captured the other atrium. The latter resulted in termination when pacing was stopped in 4 of these 6 experiments. Propafenone did not change the duration of focal AF before GP ablation (17+/-9 versus 14+/-8 minutes; control, P=0.6) but terminated reentrant atrial tachyarrhythmias (12+/-3 versus 2+/-1 minutes, P=0.0009). CONCLUSIONS: Before GP ablation, acetylcholine (100 mmol/L) induced sustained AF characterized by rapid, focal firing. GP ablations were associated with loss of focal firing and regularization of electrograms in both atria before termination. Propafenone failed to terminate focal AF but rapidly terminated entrainable macroreentrant atrial tachyarrhythmias. PMID- 19808495 TI - Long-term outcomes after catheter ablation of cavo-tricuspid isthmus dependent atrial flutter: a meta-analysis. AB - BACKGROUND: Despite the success of catheter ablation of cavotricuspid isthmus dependent atrial flutter (AFL), important postablation outcomes are ill-defined. The purpose of our study was to analyze long-term outcomes after catheter ablation of cavotricuspid isthmus-dependent AFL. METHODS AND RESULTS: A meta analysis was performed of articles reporting clinical outcomes after catheter ablation of AFL published between January 1988 and July 2008. The analysis included 158 studies comprising 10 719 patients (79% men, 59.8+/-0.5 years old, 46% left atrial enlargement, 46% heart disease, 42% with history of atrial fibrillation, 14.3+/-0.4 months of follow-up). The overall acute success rate adjusted for reporting bias was 91.1% (95% CI, 89.5 to 92.4), 92.7% (95% CI, 90.0 to 94.8) for 8- to 10-mm tip/or irrigated radiofrequency catheters, and 87.9% (95% CI, 84.2 to 90.9) for 4- to 6-mm tip catheters (P>0.05). Atrial flutter recurrence rates were significantly reduced by use of 8- to 10-mm tip or irrigated radiofrequency catheters (6.7% versus 13.8%, P<0.05) and by use of bidirectional cavotricuspid isthmus block as a procedural end point (9.3% versus 23.6%, P<0.05). The AFL recurrence rate did not increase over time. The overall occurrence rate of atrial fibrillation after AFL ablation was 33.6% (95% CI, 29.7 to 37.3) but was 52.7% (95% CI, 47.8 to 57.6) in patients with a history of atrial fibrillation before ablation and 23.1% (95% CI, 17.5 to 29.9) in those without atrial fibrillation before ablation (P<0.05). The incidence of atrial fibrillation increased over time in both groups; however, 5 years after ablation, the incidence of atrial fibrillation was similar in those with and without atrial fibrillation before ablation. The acute complication rate was 2.6% (95% CI, 2 to 3). The mortality rate during follow-up was 3.3% (95% CI, 2.4 to 4.5). Antiarrhythmic drug use after ablation was 31.6% (95% CI, 25.6 to 37.8). The long term use of coumadin was 65.9%, (95% CI, 43.8 to 82.8). Quality of life data were very limited. CONCLUSIONS: AFL ablation is safe and effective. Ablation technology and procedural end points have greater influences on AFL recurrences than on acute ablation success rates. Atrial fibrillation is common after AFL ablation. Almost one third of patients take antiarrhythmic drugs after AFL ablation. Atrial fibrillation before AFL ablation may indicate a more advanced state of electric disease. PMID- 19808496 TI - Phrenic stimulation: a challenge for cardiac resynchronization therapy. AB - BACKGROUND: Phrenic stimulation (PS) may hinder left ventricular (LV) pacing. We prospectively observed its prevalence in consecutive patients with cardiac resynchronization therapy (CRT) devices. METHODS AND RESULTS: In the years 2003 to 2006, 197 patients received a CRT device. PS and LV threshold measurements were carried out at implantation and at 6-month follow-up. LV reverse remodeling was assessed by echocardiography before implantation and at follow-up. LV lead placement was lateral/posterolateral in 86% of patients. Both PS and LV reverse remodeling occurred most frequently at the lateral/posterolateral LV pacing sites (P<0.001). PS was detected in 73 (37%) of patients and was clinically relevant in 41 (22%). The detection of PS at implantation had a poor sensitivity, as it occurred only in left lateral or sitting position in 27 patients. Ten patients (5%) underwent repeated surgery and 4 (2%) had their CRT turned off because of PS. At follow-up, we could manage PS noninvasively in 32 patients with a small PS LV threshold difference: in 20 by cathode programmability (3 also thanks to automatic management of LV output) and in 12 (without cathode programmability) by programming the LV output as threshold +1 V. CONCLUSIONS: PS may seriously hinder CRT. A bipolar LV lead and cathode programmability are mandatory to avoid PS by changing the LV pacing vector at target sites for CRT. LV stability at target sites despite PS should also be pursued by these means. The automatic adjustment of LV pacing output is complementary in patients with a small PS-LV threshold difference. PMID- 19808497 TI - Risk of failure of transvenous implantable cardioverter-defibrillator leads. AB - BACKGROUND: Despite the positive effect on mortality in selected patients, implantable cardioverter-defibrillator therapy is also associated with potential malfunction of the implanted system. The present study provides the long-term lead failure rate in a large single-center cohort. METHODS AND RESULTS: Since 1992, a total of 2068 implantable cardioverter-defibrillator patients with 2161 defibrillation leads were prospectively collected. Data of the implant procedure and all follow-up visits were recorded. All cases of lead removal or capping or placing of an additional pace or sense lead were noted and analyzed. Lead models were grouped by manufacturer and approximate lead diameter in French. During a mean follow-up of 36 months, 82 (3.8%) cases of lead failure were identified. Cumulative incidence of lead failure at 1 year was 0.6%; at 5 years, 6.5%; and at 10 years, 16.4%. The highest risk of lead failure was found in small-diameter leads. Adjusted hazard ratio was 6.4 (95% CI, 3.2 to 12.8) for Medtronic 7F leads, when compared with all other leads. CONCLUSIONS: In this large single center experience, the overall incidence of lead failure was 1.3 (95% CI, 1.0 to 1.6) per 100 lead-years. Comparison of different groups of leads shows major differences in event rates. Specific manufacturer's small-diameter defibrillation leads may have a higher risk of early lead failure. PMID- 19808498 TI - Biophysical properties of 9 KCNQ1 mutations associated with long-QT syndrome. AB - BACKGROUND: Inherited long-QT syndrome is characterized by prolonged QT interval on the ECG, syncope, and sudden death caused by ventricular arrhythmia. Causative mutations occur mostly in cardiac potassium and sodium channel subunit genes. Confidence in mutation pathogenicity is usually reached through family genotype phenotype tracking, control population studies, molecular modeling, and phylogenetic alignments; however, biophysical testing offers a higher degree of validating evidence. METHODS AND RESULTS: By using in vitro electrophysiological testing of transfected mutant and wild-type long-QT syndrome constructs into Chinese hamster ovary cells, we investigated the biophysical properties of 9 KCNQ1 missense mutations (A46T, T265I, F269S, A302V, G316E, F339S, R360G, H455Y, and S546L) identified in a New Zealand-based long-QT syndrome screening program. We demonstrate through electrophysiology and molecular modeling that 7 of the missense mutations have profound pathological dominant-negative loss-of-function properties, confirming their likely disease-causing nature. This supports the use of these mutations in diagnostic family screening. Two mutations (A46T, T265I) show suggestive evidence of pathogenicity within the experimental limits of biophysical testing, indicating that these variants are disease-causing via delayed- or fast-activation kinetics. Further investigation of the A46T family has revealed an inconsistent cosegregation of the variant with the clinical phenotype. CONCLUSIONS: Electrophysiological characterization should be used to validate long-QT syndrome pathogenicity of novel missense channelopathies. When such results are inconclusive, great care should be taken with genetic counseling and screening of such families, and alternative disease-causing mechanisms should be considered. PMID- 19808499 TI - Clinical correlates and heritability of QT interval duration in blacks: the Jackson Heart Study. AB - BACKGROUND: Electrocardiographic QT interval prolongation is a risk factor for sudden cardiac death and drug-induced arrhythmia. The clinical correlates and heritability of QT interval duration in blacks have not been well studied despite their higher risk for sudden cardiac death compared with non-Hispanic whites. We sought to investigate potential correlates of the QT interval and estimate its heritability in the Jackson Heart Study. METHODS AND RESULTS: The Jackson Heart Study comprises a sample of blacks residing in Jackson, Miss, of whom 5302 individuals with data at the baseline examination were available for study. Jackson Heart Study participants on QT-altering medications, with bundle-branch block, paced rhythm, atrial fibrillation/flutter, or other arrhythmias were excluded, resulting in a sample of 4660 individuals eligible for analyses. The relation between QT and potential covariates was tested using multivariable stepwise linear regression. Heritability was estimated using Sequential Oligogenic Linkage Analysis Routine in a subset of 1297 Jackson Heart Study participants in 292 families; the remaining sample included unrelated individuals. In stepwise multivariable linear regression analysis, covariates significantly associated with QT interval duration included R-R interval, sex, QRS duration, age, serum potassium, hypertension, body mass index, coronary heart disease, diuretic use, and Sokolow-Lyon voltage (P < or = 0.01 for all). The heritability of QT interval duration in the age-, sex-, and R-R interval-adjusted model and in the fully adjusted model was 0.41 (SE, 0.07) and 0.40 (SE, 0.07; P < 10(-11) for both), respectively. CONCLUSIONS: There is substantial heritability of adjusted QT interval in blacks, supporting the need for further investigation to identify its genetic determinants. PMID- 19808500 TI - Three distinct directions of intramural activation reveal nonuniform side-to-side electrical coupling of ventricular myocytes. AB - BACKGROUND: The anisotropy of cardiac tissue is a key determinant of 3D electric propagation and the stability of activation wave fronts in the heart. The electric properties of ventricular myocardium are widely assumed to be axially anisotropic, with activation propagating most rapidly in the myofiber direction and at uniform velocity transverse to this. We present new experimental evidence that contradicts this view. METHODS AND RESULTS: For the first time, high-density intramural electric mapping (325 electrodes at approximately 4x4x1-mm spacing) from pig left ventricular tissue was used to reconstruct 3D paced activation surfaces projected directly onto 3D tissue structure imaged throughout the same left ventricular volume. These data from 5 hearts demonstrate that ventricular tissue is electrically orthotropic with 3 distinct propagation directions that coincide with local microstructural axes defined by the laminar arrangement of ventricular myocytes. The maximum conduction velocity of 0.67+/-0.019 ms(-1) was aligned with the myofiber axis. However, transverse to this, the maximum conduction velocity was 0.30+/-0.010 ms(-1), parallel to the myocyte layers and 0.17+/-0.004 ms(-1) normal to them. These orthotropic conduction velocities give rise to preferential activation pathways across the left ventricular free wall that are not captured by structurally detailed computer models, which incorporate axially anisotropic electric properties. CONCLUSIONS: Our findings suggest that current views on uniform side-to-side electric coupling in the heart need to be revised. In particular, nonuniform laminar myocardial architecture and associated electric orthotropy should be included in future models of initiation and maintenance of ventricular arrhythmia. PMID- 19808501 TI - Simultaneous biventricular noncontact mapping and ablation of septal ventricular tachycardia in a chronic ovine infarct model. AB - BACKGROUND: We assessed a novel simultaneous biventricular mapping and ablation approach for septal ventricular tachycardia (VT) in a chronic ovine infarct model. METHODS AND RESULTS: In 8 sheep with inducible VT, mapping and ablation were performed 9+/-3 months after percutaneously induced myocardial infarction, with left ventricular ejection fraction 23+/-8%. Scar was identified by EnSite Dynamic Substrate Mapping plus CARTO voltage mapping. Thirty VT episodes (cycle length, 235+/-42 ms) were mapped with simultaneous analyses using EnSite arrays deployed in both the left ventricle and the right ventricle. Short ablation lines were created perpendicular to the breakout pathway along the scar border in the ventricle with earliest activity. If septal VT was still inducible, this line was extended before ablation in the second chamber. The end point of noninducibility of VT was achieved in all animals. The mean difference in delay in noncontact breakout timing between the ventricles was shorter for VT with (n=18) than without (n=12) septal breakout (32+/-7.8 ms, P<0.001). In 5 of 6 animals, after ablation in one ventricle, septal VT was still inducible with a common breakout site in the second ventricle. After septal ablation in the second ventricle, VT was no longer inducible. In the 6 animals in which septal VT had been ablated, transmural septal ablation was identified at the scar border, with overlapping left ventricular and right ventricular ablation lesions present in 5 of 6 (septal thickness 8 to 17 mm) and left ventricular endocardial ablation being transmural in 1 of 6 (6 mm). CONCLUSIONS: Biventricular scar and VT activation mapping correctly localizes septal VT pathways, directing ablation from one or both septal endocardial aspects. Creation of a transmural septal lesion at the scar border interrupting VT exit points is highly effective at ablating septal VT. PMID- 19808502 TI - Impact of sleep on arrhythmogenesis. PMID- 19808504 TI - Malignant presentation of cardiac hemangioma: a rare cause of complete atrioventricular block. PMID- 19808503 TI - Impact of dietary fatty acids on cardiac arrhythmogenesis. PMID- 19808505 TI - Coarctation of the abdominal aorta: an uncommon cause of arterial hypertension and stroke. PMID- 19808506 TI - Catastrophic coronary stent fracture and coronary perforation presenting as cardiogenic shock: a rare but fatal late complication of stenting. PMID- 19808508 TI - In search of a holy grail: predicting cardiac resynchronization therapy outcomes by echocardiography. PMID- 19808509 TI - Multimodality evaluation of the viability of stem cells delivered into different zones of myocardial infarction. AB - BACKGROUND: We tested the hypothesis that multimodality imaging of mouse embryonic stem cells (mESCs) provides accurate assessment of cellular location, viability, and restorative potential after transplantation into different zones of myocardial infarction. METHODS AND RESULTS: Mice underwent left anterior descending artery ligation followed by transplantation of dual-labeled mESCs with superparamagnetic iron oxide and luciferase via direct injection into 3 different zones of myocardial infarction: intra-infarction, peri-infarction, and normal (remote). One day after transplantation, magnetic resonance imaging enabled assessment of the precise anatomic locations of mESCs. Bioluminescence imaging allowed longitudinal analysis of cell viability through detection of luciferase activity. Subsequent evaluation of myocardial regeneration and functional restoration was performed by echocardiography and pressure-volume loop analysis. Using 16-segment analysis, we demonstrated precise localization of dual-labeled mESCs. A strong correlation between histology and magnetic resonance imaging was established (r=0.962, P=0.002). Bioluminescent imaging data demonstrated that cell viability in the remote group was significantly higher than in other groups. Echocardiography and pressure-volume loop analysis revealed improved functional restoration in animals treated with mESCs, although myocardial regeneration was not observed. CONCLUSIONS: Multimodality evaluation of mESC engraftment in the heterogeneous tissue of myocardial infarction is possible. Magnetic resonance imaging demonstrated accurate anatomic localization of dual-labeled mESCs. Bioluminescent imaging enabled assessment of variable viability of mESCs transplanted into the infarcted myocardium. Echocardiography and pressure-volume loop analysis validated the restorative potential of mESCs. Although mESCs transplanted into the remote zone demonstrated the highest viability, precise delivery of mESCs into the peri-infarction region might be equally critical in restoring the injured myocardium. PMID- 19808510 TI - Strain dyssynchrony index correlates with improvement in left ventricular volume after cardiac resynchronization therapy better than tissue velocity dyssynchrony indexes. AB - BACKGROUND: Various dyssynchrony indexes derived from tissue velocity and strain imaging have been proposed to predict the effectiveness of cardiac resynchronization therapy (CRT). We sought to compare the effect of CRT on dyssynchrony indexes derived by tissue velocity and strain and to determine which baseline intraventricular dyssynchrony parameters correlate with improvement in left ventricular volume after CRT. METHODS AND RESULTS: Echocardiography with tissue Doppler imaging was performed in 45 patients with systolic heart failure at baseline, 1 day after CRT, and a median of 6 months after CRT. We calculated septal-lateral delay and anteroseptal-posterior delay and standard deviation of time to peak systolic velocity in the 12 basal and mid-left ventricular segments (Tv-SD). The standard deviation for time to peak strain in the 12 basal and mid left ventricular segments (Tepsilon-SD) was calculated as a strain-derived dyssynchrony index. None of the tissue velocity-derived dyssynchrony indexes improved after CRT (septal-lateral delay, P=0.39; anteroseptal-posterior delay, P=0.46; Tv-SD, P=0.30), whereas Tepsilon-SD decreased significantly after CRT (P<0.001). Improvement in Tepsilon-SD 1 day after CRT correlated with the reduction in end-systolic volume at follow-up (r=0.66; P<0.001). Baseline Tepsilon-SD demonstrated significant correlation with the reduction of end systolic volume at follow-up (r=0.57; P<0.001); however, baseline tissue velocity derived dyssynchrony indexes failed to predict the effect of CRT. CONCLUSIONS: The strain-derived dyssynchrony index is a better measurement than the tissue velocity dyssynchrony index for monitoring changes in mechanical dyssynchrony after CRT and for predicting reduction in left ventricular volume after CRT. PMID- 19808511 TI - Magnetic resonance-derived 3-dimensional blood flow patterns in the main pulmonary artery as a marker of pulmonary hypertension and a measure of elevated mean pulmonary arterial pressure. AB - BACKGROUND: Pulmonary hypertension is a disease characterized by an elevation in pulmonary arterial pressure that is diagnosed invasively via right heart catheterization. Such pathological altered pressures in the pulmonary vascular system should lead to changes in blood flow patterns in the main pulmonary artery. METHODS AND RESULTS: Forty-eight subjects (22 with manifest pulmonary hypertension, 13 with latent pulmonary hypertension, and 13 normal control subjects) underwent time-resolved 3D magnetic resonance phase-contrast imaging of the main pulmonary artery. Velocity fields that resulted from measurements were calculated, visualized, and analyzed with dedicated software. Main findings were as follows: (1) Manifest pulmonary hypertension coincides with the appearance of a vortex of blood flow in the main pulmonary artery (sensitivity and specificity of 1.00, 95% confidence intervals of 0.84 to 1.00 and 0.87 to 1.00, respectively), and (2) the relative period of existence of the vortex correlates significantly with mean pulmonary arterial pressure at rest (correlation coefficient of 0.94). To test the diagnostic performance of the vortex criterion, we furthermore investigated 55 patients in a blinded prospective study (22 with manifest pulmonary hypertension, 32 with latent pulmonary hypertension, and 1 healthy subject), which resulted in a sensitivity of 1.00 and specificity of 0.91 (95% confidence intervals of 0.84 to 1.00 and 0.76 to 0.98, respectively). Comparison of catheter-derived mean pulmonary artery pressure measurements and calculated mean pulmonary artery pressure values resulted in a standard deviation of differences of 3.6 mm Hg. CONCLUSIONS: Vortices of blood flow in the main pulmonary artery enable the identification of manifest pulmonary hypertension. Elevated mean pulmonary arterial pressures can be measured from the period of vortex existence. PMID- 19808512 TI - Direct en face imaging of secundum atrial septal defects by velocity-encoded cardiovascular magnetic resonance in patients evaluated for possible transcatheter closure. AB - BACKGROUND: Atrial septal defect (ASD) flow can be measured indirectly by velocity-encoded cardiovascular magnetic resonance (veCMR) of the pulmonary artery and aorta. Imaging the secundum ASD en face could potentially enable direct flow measurement and provide valuable information about ASD size, shape, location, and proximity to other structures. METHODS AND RESULTS: Forty-four patients referred for possible transcatheter ASD closure underwent a comprehensive standard evaluation, including transesophageal and/or intracardiac echocardiography and invasive oximetry. CMR was performed in parallel and included direct en face veCMR after an optimal double-oblique imaging plane was determined that accounted for ASD flow direction and cardiac-cycle interatrial septal motion. ASD flow measured by direct en face veCMR correlated better with invasive oximetry than indirect (pulmonary artery and aorta) veCMR (r=0.80 versus r=0.66). Additionally, 95% limits of agreement were narrower (+/-3.9 versus +/ 5.1 L/min). En face veCMR determined that defects usually were eccentrically shaped (major/minor axis length >1.5) rather than circular, with 16% having extreme eccentricity (major/minor >2.0). Overall, ASD size by both veCMR and intracardiac echocardiography correlated with final device size; however, in small to medium defects (<3 cm(2)) and extremely eccentric defects, veCMR correlated better with final device size than did intracardiac echocardiography. Importantly, CMR identified additional information in 9 patients (20%) that altered clinical management. Specifically, en face veCMR detected additional defects (n=3), large ASD with insufficient rim tissue (n=2), and sinus venosus defect with anomalous pulmonary vein (n=1). Cine and/or morphological imaging detected interrupted inferior vena cava (n=2) and sinus of Valsalva aneurysm (n=1). CONCLUSIONS: En face veCMR with an optimized imaging plane can determine ASD flow, size, and morphology. CMR provided information incremental to comprehensive standard evaluation that altered clinical management in 20% of patients. PMID- 19808513 TI - Tomographic left ventricular volumetric emptying analysis by real-time 3 dimensional echocardiography: influence of left ventricular dysfunction with and without electrical dyssynchrony. AB - BACKGROUND: The sequence of left ventricular (LV) systolic emptying is not completely understood. Using real-time 3-dimensional echocardiography, we investigated this sequence and LV synchronicity in physiological and pathological conditions. METHODS AND RESULTS: The study population consisted of 116 healthy volunteers, 20 top-level athletes, 35 patients with LV dysfunction, and 84 patients with LV dysfunction and left bundle-branch block (LBBB). We subdivided the LV into 16 volumetric segments for regional analysis and into apical, middle, and basal regions to calculate the mean of end-systolic times and the time to minimum systolic volume of each region. In healthy volunteers and in top-level athletes, the emptying systolic times increased smoothly from apex to base. These differences determined an apex-to-base time gradient in the LV emptying sequence. In patients with LV dysfunction and without LBBB, this gradient was maintained with a relatively higher LV dyssynchrony. However, in patients with LV dysfunction and LBBB, there was no clear sequence in LV emptying volumes, and this group had the highest LV dyssynchrony. CONCLUSIONS: Real-time 3-dimensional echocardiography tomographic slicing of the LV enables accurate analysis of LV emptying in physiological conditions and in conditions of LV dysfunction with and without electrical dyssynchrony. Progressive dilation of LV produces deterioration in LV synchronicity. However, it is the presence of LV dysfunction in combination with LBBB that determines the loss of the apex-to-base time gradient in LV emptying. PMID- 19808514 TI - Left ventricular diastolic mechanical dyssynchrony and associated clinical outcomes in children with dilated cardiomyopathy. AB - BACKGROUND: We investigated diastolic mechanical dyssynchrony and its relation to clinical status in pediatric dilated cardiomyopathy (DCM). METHODS AND RESULTS: We calculated a diastolic and systolic dyssynchrony index (standard deviation of time to peak tissue early diastolic/systolic velocity in 12 left ventricular segments) in 33 children with DCM and 46 control subjects. A threshold to diagnose diastolic dyssynchrony was determined, and cardiac function and clinical outcomes were compared between DCM patients with and without diastolic dyssynchrony. Left ventricular wall motion was more synchronized in diastole than in systole. The diastolic dyssynchrony index was significantly higher in children with DCM than in control subjects (28.1+/-18.1 versus 9.1+/-3.8 ms, P<0.0001). A 17-ms threshold indicated the presence of diastolic dyssynchrony. Patients who died or underwent transplantation had greater diastolic dyssynchrony (diastolic dyssynchrony index 37.9+/-20.5 versus 22.1+/-13.8 ms, P=0.01), and the rate of transplant-free survival appeared to be worse for DCM patients with diastolic dyssynchrony than for patients with synchronous DCM (hazard ratio 2.98, P=0.11; hazard ratio adjusted for disease duration 2.95, P=0.17). CONCLUSIONS: Left ventricular diastolic mechanical dyssynchrony is common in pediatric DCM, especially in patients who subsequently experience transplantation or death, and may be associated with a decreased length of transplantation-free survival. PMID- 19808515 TI - The future of cardiovascular imaging in the diagnosis and management of heart failure, part 1: tasks and tools. PMID- 19808516 TI - Is echocardiographic assessment of dyssynchrony useful to select candidates for cardiac resynchronization therapy? Echocardiography is not useful before cardiac resynchronization therapy if QRS duration is available. PMID- 19808517 TI - Is echocardiographic assessment of dyssynchrony useful to select candidates for cardiac resynchronization therapy? Echocardiography is useful before cardiac resynchronization therapy if QRS duration is available. PMID- 19808518 TI - Real-time 3-dimensional transesophageal echocardiography during left atrial radiofrequency catheter ablation for atrial fibrillation. PMID- 19808519 TI - Tissue characterization of a suspected aortic valve fibroelastoma with cardiac magnetic resonance imaging. PMID- 19808520 TI - Paradoxical systemic embolization in hereditary hemorrhagic telangiectasia. PMID- 19808521 TI - An unusual cause of aortic regurgitation. PMID- 19808522 TI - Percutaneous closure of multiple secundum atrial septal defects using 3 amplatzer atrial septal occluder devices: evaluation by live transthoracic 3-dimensional echocardiography. PMID- 19808523 TI - Paradoxical embolism via a patent foramen ovale: an important mechanism of cryptogenic strokes. PMID- 19808524 TI - Noninvasive coronary imaging: the contest between magnetic resonance and computed tomographic coronary angiography. PMID- 19808525 TI - Imaging the physiology of the ischemic cascade: are 2 tools better than 1? PMID- 19808526 TI - Serial noninvasive in vivo positron emission tomographic tracking of percutaneously intramyocardially injected autologous porcine mesenchymal stem cells modified for transgene reporter gene expression. AB - BACKGROUND: Porcine bone marrow-derived mesenchymal stem cells (MSCs) were stably transfected with a lentiviral vector for transgene expression of the trifusion protein renilla luciferase, red fluorescent protein and herpes simplex truncated thymidine kinase (LV-RL-RFP-tTK; positron emission tomography [PET] reporter gene) for in vivo noninvasive tracking of the intramyocardially delivered MSC fate. METHODS AND RESULTS: A closed-chest, reperfused myocardial infarction was created in farm pigs. Sixteen days after myocardial infarction, LV-RL-RFP-tTK MSCs were injected intramyocardially using electromechanical mapping guidance in the infarct border zone (n=7). PET-computed tomographic metabolic and perfusion imaging was performed after an intravenous injection of 10 mCi [18F]-FHBG and 13N ammonia PET at 30+/-2 hours and 7 days after LV-RL-RFP-tTK-MSC treatment. Fusion imaging of the [18F]-FHBG PET-computed tomography with MRI was used to determine the myocardial location of the injected LV-RL-RFP-tTK-MSCs. Seven days after injections, [18F]-FHBG PET showed a decreased cardiac uptake with a mild increased pericardial and pleura uptake in the treated animals, which was confirmed by the measurement of luciferase activity. At 10 days, infarct size by MRI in the LV-RL-RFP-tTK-MSC-treated animals was smaller than controls (n=7) (23.3+/-1.5% versus 30.2+/-3.5%, P<0.005). The presence of the LV-RL-RFP-tTK-MSCs (5.8+/-1.1% of the injected cells) in the myocardium 10 days after intramyocardial delivery was confirmed histologically. CONCLUSIONS: Reporter gene imaging enables the tracking of the persistence of viable LV-RL-RFP-tTK-MSC in the peri-infarcted porcine myocardium at 10 days after delivery using clinical PET scanners. PMID- 19808527 TI - Aortic root measurement by cardiovascular magnetic resonance: specification of planes and lines of measurement and corresponding normal values. AB - BACKGROUND: Cardiovascular magnetic resonance is widely used for aortic root visualization and measurement, but methods still need to be standardized. Our aim was to identify appropriate planes of acquisition and lines of measurement and record corresponding normal values. METHODS AND RESULTS: We studied 120 healthy volunteers, 10 of each gender in each decile between 20 and 80 years, by using a 1.5-T cardiovascular magnetic resonance system. Steady-state free precession cine acquisitions aligned with the left ventricular outflow tract in oblique sagittal and coronal orientations were used to locate 2 sinus planes that transected the root at its widest point in its maximally expanded systolic and at its end diastolic positions. We measured the cusp-cusp and the cusp-commissure dimensions in these cine planes, each as the average of 3. Diastolic cusp-commissure dimensions were smaller than diastolic cusp-cusp dimensions (32.0+/-3.5 mm versus 34.6+/-4.0 mm in men, 28.4+/-2.8 mm versus 30.7+/-3.3 mm in women, P<0.001 for both). The diastolic cusp-commissure dimensions increased by 0.9 mm per decade in men and 0.7 mm per decade in women (P<0.001 for both) and gave higher R(2) values with respect to age and body surface area (0.40 for men, 0.27 for women) than diastolic cusp-cusp, systolic cusp-commissure, or sinus measurements made in the left ventricular outflow tract planes. CONCLUSIONS: The results indicate the importance of consistent methods for measurement of the aortic root by cardiovascular magnetic resonance. We recommend diastolic cusp-commissure measurements, which yielded favorable R(2) values with respect to age and body surface area and were found to correspond closely with reference echocardiographic root measurements recorded in the Framingham cohort. We recorded reference values for these and other possible aortic root measurements by cardiovascular magnetic resonance. PMID- 19808528 TI - Direct comparison of whole-heart navigator-gated magnetic resonance coronary angiography and 40- and 64-slice multidetector row computed tomography to detect the coronary artery stenosis in patients scheduled for conventional coronary angiography. AB - BACKGROUND: Both whole-heart magnetic resonance coronary angiography (WH-MRCA) and multidetector computed tomography (MDCT) have been proposed for the noninvasive identification of the coronary stenosis. The authors sought to directly compare the diagnostic accuracy of these noninvasive imaging techniques using the invasive quantitative coronary angiography as a reference standard. METHODS AND RESULTS: Seventy-seven consecutive patients (56 men, 61+/-14 years) prospectively underwent WH-MRCA and 40- or 64-slice MDCT before the quantitative coronary angiography. Diagnostic accuracy of WH-MRCA and MDCT for the visual identification of >50% diameter stenosis in segments of >1.5 mm size was compared using the quantitative coronary angiography as a reference. According to the quantitative coronary angiography, 49 of 992 coronary segments >1.5 mm diameter had >50% diameter stenosis. MDCT had a higher success rate (100% versus 88%, P<0.001) and enabled identification of more segments (963 versus 726, P<0.001) than did WH-MRCA. On a per-segment basis, WH-MRCA had similar sensitivity (47/49 or 96% versus 48/49 or 98%, P=0.9) but significantly lower specificity (644/943 or 68% versus 863/943 or 92%, P<0.001) and accuracy (691/992 or 70% versus 911/992 or 92%, P<0.001) for the detection of >50% diameter stenosis than did MDCT. On a per-patient basis, the sensitivity was similar (17/17 or 100% versus 16/17 or 94%, P=0.9), but specificity (43/60 or 72% versus 53/60 or 88%, P=0.024) and diagnostic accuracy (60/77 or 78%, versus 69/77 or 90%, P=0.044) of WH-MRCA for the detection of >50% diameter stenosis were significantly lower than of MDCT. CONCLUSIONS: Because of the higher success rate and higher number of interpretable segments, 40- or 64-slice MDCT performs better than WH-MRCA. PMID- 19808529 TI - Additional value of myocardial perfusion imaging during dobutamine stress magnetic resonance for the assessment of coronary artery disease. AB - BACKGROUND: Dobutamine stress magnetic resonance (DSMR) imaging has emerged as a valuable tool for the detection of inducible wall motion abnormalities. The role of perfusion imaging during DSMR is not well defined. We examined whether the addition of myocardial perfusion imaging during DSMR provides incremental benefit for the evaluation of coronary artery disease. METHODS AND RESULTS: DSMR was combined with perfusion imaging in 455 consecutive patients who were scheduled for clinically indicated invasive coronary angiography. Perfusion images were acquired in 3 standard short-views at rest and during maximum dobutamine-atropine stress. Wall motion and perfusion images were interpreted sequentially, blinded to other data. Significant (> or =70%) stenoses were present in 285 patients on invasive coronary angiography. The use of DSMR combined with perfusion imaging versus DSMR increased sensitivity (91% versus 85%, P=0.001), but not specificity (70% versus 82%, P=0.001), resulting in identical overall diagnostic accuracy (84% versus 84%, P=NS; Youden index 0.61 versus 0.67). DSMR combined with perfusion imaging enabled the correct diagnosis of coronary artery disease in an additional 13% of DSMR-negative patients at the cost of 11% more false-positive cases. CONCLUSIONS: The addition of perfusion imaging during DSMR improves sensitivity for the diagnosis of coronary artery disease but does not enhance overall diagnostic accuracy because of a concomitant decrease in specificity. PMID- 19808530 TI - 123 I-mIBG scintigraphy to predict inducibility of ventricular arrhythmias on cardiac electrophysiology testing: a prospective multicenter pilot study. AB - BACKGROUND: Disturbances of autonomic function after infarction are associated with both total mortality and sudden death. Although many imaging techniques for assessing the cardiac autonomic nervous system have been studied, the clinical usefulness of these techniques remains uncertain. This exploratory pilot study examined the relationship between abnormalities of ventricular sympathetic innervation delineated by scintigraphic imaging with (123)I-mIBG and inducible ventricular tachyarrhythmias in patients with left ventricular dysfunction and previous myocardial infarction. METHODS AND RESULTS: Fifty patients underwent electrophysiological (EP) testing and 15-minute and 4-hour planar and single photon emission computed tomography (SPECT) imaging with (123)I-mIBG and SPECT imaging with (99m)Tc-tetrofosmin. The primary efficacy variables were the 4-hour heart:mediastinum ratio (H/M) and the (123)I-mIBG/(99m)Tc-tetrofosmin SPECT mismatch score. EP studies were categorized as positive (EP(+)) or negative (EP( )) for inducibility of sustained (>30 seconds) ventricular tachyarrhythmias. Thirty patients were EP(+), and 20 were EP(-). There were no significant differences in the 4-hour H/M ratios or (123)I-mIBG/(99m)Tc-tetrofosmin SPECT mismatch scores between the two groups. In a multivariable analysis using all (123)I-mIBG and (99m)Tc-tetrofosmin SPECT measurements, the only variable that showed a significant difference between EP(+) and EP(-) patients was the 4-hour (123)I-mIBG SPECT defect score. A 4-hour (123)I-mIBG SPECT defect score of > or =37 yielded a sensitivity of 77% and specificity of 75% for predicting EP results. CONCLUSIONS: The standard indices of (123)I-mIBG imaging (H/M and innervation-perfusion mismatch score) are not predictive of EP test results. The association of (123)I-mIBG SPECT defect severity with EP test inducibility in this exploratory study will require confirmation in a larger cohort of patients. PMID- 19808531 TI - Functional analysis of the components of the right ventricle in the setting of tetralogy of Fallot. AB - BACKGROUND: Anatomic and functional observations suggest that the right ventricle (RV) can be analyzed in terms of its inlet, apical trabecular, and outlet components. Our study was designed to evaluate the regional adaptation of these components to different conditions of loading, with additional analysis of the surgical techniques used for primary repair. METHODS AND RESULTS: We studied prospectively 45 patients with tetralogy of Fallot (age, 20.5+/-8.1 years) and 24 control subjects (age, 20.1+/-5.8 years). All subjects were studied by using cardiac MRI. End-diastolic (EDV), end-systolic (ESV), stroke volumes (SV), and ejection fraction (EF) were determined for the overall RV and separately for its inlet, apical trabecular, and outlet components. The patients had pulmonary regurgitant fractions of 33.2+/-11.1%, and RV peak-systolic pressures of 40.7+/ 16.1 mm Hg. In controls, the apical trabecular component EDV was 51.5+/-11.1 mL/m(2) (54.3+/-6.8% of the total RV EDV), ESV was 19.2+/-6.3 mL/m(2) (47.6+/ 10.5% of RV ESV), and SV was 32.3+/-6.9 mL/m(2) (58.9+/-6.6% of RV SV), resulting in an EF of 63.1+/-7.7%. When considering all patients, the apical trabecular component took up the greatest part of the overload, having an EDV of 76.5+/-18.1 mL/m(2), and an ESV of 31.6+/-12.8 mL/m(2), reflecting an increase of 49 and 67% over controls, respectively (P<0.001). EF was 59.7+/-10.7%, and was maintained at control levels (P=0.132). In controls, the outlet had considerable ejecting force, with an EF of 54.8+/-9.1%, whereas it was decreased in the patients with tetralogy (EF=28.5+/-11.9%). There was significant increase of ESV (P<0.001), but not of EDV, with EF decreased by 45% (P<0.001). The inlet was not significantly affected by overload. The surgical technique did not significantly affect any measured parameter for any component. CONCLUSIONS: In patients with tetralogy of Fallot, subsequent to surgical correction, the individual components of the RV respond in characteristic fashion to RV overload. PMID- 19808532 TI - Cardiovascular magnetic resonance for direct assessment of anatomic regurgitant orifice in mitral regurgitation. AB - BACKGROUND: In patients with mitral regurgitation (MR), assessment of the severity of valvular dysfunction is crucial. Recently, regurgitant orifice area has been proposed as the most useful indicator of the severity of MR. The purpose of our study was to determine whether planimetry of the anatomic regurgitant orifice (ARO) in patients with MR is feasible by cardiovascular magnetic resonance (CMR) and correlates with invasive catheterization and echocardiography effective regurgitant orifice [ECHO-ERO] by proximal isovelocity surface area. METHODS AND RESULTS: Planimetry of ARO was performed with a 1.5-T CMR scanner using a breath-hold balanced gradient echo sequence true fast imaging with steady state precession (TrueFISP). CMR planimetry of ARO was possible in 35 of 38 patients and was closely correlated with angiographic grading (r=0.84, P<0.0001). In patients with MR grade > or =III on catheterization, CMR-ARO (0.60+/-0.29 cm(2) versus 0.30+/-0.19 cm(2), P<0.0001) as well as ECHO-ERO (0.49+/-0.17 cm(2) versus 0.27+/-0.10 cm(2)) were significantly elevated in comparison with MR grade or =III as defined by catheterization, with a sensitivity and specificity of 94% and 94%, respectively. CONCLUSIONS: CMR planimetry of the anatomic mitral regurgitant lesion in patients with MR is feasible and permits quantification of MR with good agreement with the accepted invasive and noninvasive methods. Direct measurement by CMR is a promising new method for the precise assessment of ARO area and the severity of MR. PMID- 19808533 TI - Retinal vascular imaging: a new tool in microvascular disease research. AB - The microcirculation is relatively inaccessible to direct visualization and investigation. Recent methods have been developed which use advanced retinal photographic imaging techniques and computer-assisted image analysis to characterize, measure and quantify subtle variations and abnormalities in the retinal vasculature. These quantitative and qualitative assessments demonstrate a close association of retinal vascular signs to both clinical and subclinical cerebrovascular, cardiovascular and metabolic outcomes. Retinal vascular imaging may thus offer potential as a noninvasive research tool to probe the role and pathophysiology of the microvasculature, and as a cardiovascular risk prediction tool. Key areas where retinal vascular imaging has contributed to increased understanding of microvascular pathology and major areas of current and new research are discussed in this review. PMID- 19808534 TI - The future of cardiovascular imaging in the diagnosis and management of heart failure, part 2: clinical applications. PMID- 19808535 TI - Subepicardial aneurysm evaluated by multiplane 2D and real-time 3D volumetric transesophageal echocardiography. PMID- 19808536 TI - Acute isolated viral pericarditis with rapid progression to constrictive pericardial disease. PMID- 19808537 TI - Coil occlusion of aortopulmonary collateral arteries before arterial switch procedure in an infant with transposition of the great arteries. PMID- 19808538 TI - Huge ascending aortic pseudoaneurysm caused by a penetrating atherosclerotic ulcer. PMID- 19808539 TI - Prediction of sudden cardiac death in patients with coronary heart disease: the challenge ahead. PMID- 19808540 TI - "Feeling the RAGE" in the atherosclerotic vessel wall. PMID- 19808541 TI - Single-photon emission computed tomography myocardial perfusion defects are associated with an increased risk of all-cause death, cardiovascular death, and sudden cardiac death. AB - BACKGROUND: Single-photon emission computed tomography myocardial perfusion imaging defects are associated with increased all-cause mortality and cardiovascular death. However, it is unknown whether single-photon emission computed tomography myocardial perfusion imaging can identify patients at increased risk of sudden cardiac death (SCD). METHODS AND RESULTS: We analyzed a cohort of 6383 patients with angiographically documented coronary artery disease who underwent single-photon emission computed tomography imaging. Cox proportional hazards modeling was used to examine the relationship between patient characteristics and SCD. Among patients who died, the median time to SCD was 2.7 years (25(th), 75(th) percentiles 0.9, 4.9, respectively). The incidence of SCD was 3.4% (n=215) over 6.1 years (25(th), 75(th) percentiles 3.7, 9.2, respectively) of follow-up. Patients with SCD had more severe heart failure symptoms, greater comorbidity (Charlson index), and higher summed stress perfusion scores (all P<0.001). After adjusting for left ventricular ejection fraction and other clinical factors in the multivariable model, the summed stress perfusion score (fixed plus reversible defects) remained significantly associated with the occurrence of SCD: summed stress perfusion score (hazard ratios per 3 U: 1.16 [95% CI, 1.08 to 1.25], P<0.001), left ventricular ejection fraction (hazard ratios per 5 U: 0.90 [95% CI, 0.85 to 0.95], P<0.001), and Charlson index (hazard ratios 1.35 [95% CI, 1.23 to 1.49], P<0.001). CONCLUSIONS: Myocardial perfusion imaging is a significant predictor of SCD and provides information independent of clinical history and left ventricular ejection fraction. Gated single-photon emission computed tomography imaging, which evaluates both myocardial perfusion and function, may represent a more effective means of risk stratification than solitary left ventricular ejection fraction determination and should be evaluated in prospective trials. PMID- 19808542 TI - Noninvasive separation of large, medium, and small myocardial infarcts in survivors of reperfused ST-elevation myocardial infarction: a comprehensive tissue Doppler and speckle-tracking echocardiography study. AB - BACKGROUND: The objective of the study was to evaluate the ability of established and new parameters of global systolic left ventricle function to estimate myocardial infarct size. Increasing infarct extent is associated with impaired prognosis in chronic ischemic heart disease. Systolic myocardial deformation is a complex 3D process that is mainly influenced by the amount and transmural distribution of viable myocardium. Speckle-tracking echocardiography (2D-STE) enables deformation assessment along the 3 main cardiac axes independent of insonation angle. METHODS AND RESULTS: Global longitudinal, circumferential, and radial strain and left ventricle twist by 2D-STE, global longitudinal strain rate and strain by tissue Doppler imaging, and left ventricle ejection fraction and wall motion score index were assessed in 40 patients 8.5+/-5.4 months after a first myocardial infarct and compared with global myocardial infarct mass assessed by contrast-enhanced MRI. Longitudinal and circumferential strain by 2D STE and longitudinal strain and strain rate by tissue Doppler imaging significantly separated medium-sized infarcts from small or large infarcts at the global level (P<0.05). All deformation indices correlated significantly with global infarct mass (P<0.01). Circumferential and longitudinal strains by 2D-STE demonstrated the best ability to identify medium-sized global myocardial infarcts. CONCLUSIONS: Circumferential and longitudinal strains by 2D-STE correlate with myocardial infarct mass and significantly differentiate among large, medium, and small myocardial infarcts. PMID- 19808543 TI - Collagen-based matrices improve the delivery of transplanted circulating progenitor cells: development and demonstration by ex vivo radionuclide cell labeling and in vivo tracking with positron-emission tomography. AB - BACKGROUND: Collagen delivery matrices have been reported to improve the results of cell therapy, but knowledge of their mechanisms of action is limited. To evaluate whether a collagen matrix improves early engraftment posttransplantation, 2-[(18)F]fluoro-2-deoxy-d-glucose ((18)F-FDG) was used to label transplanted circulating progenitor cells (CPCs) and track them in vivo with positron-emission tomography. METHODS AND RESULTS: Efficiency of (18)F-FDG cell labeling was CPC-concentration dependent (r=0.61, P<0.001) but not (18)F-FDG dose dependent. Labeled human CPCs (2x10(6)) were injected with or without a collagen-based matrix in the ischemic hind limb of rats (n=12 per group) 2 weeks after femoral artery ligation. Imaging of labeled cells, acquired by small animal positron-emission tomography at 150 minutes postinjection, revealed greater CPC retention in the ischemic hind limb and less nonspecific leakage to other tissues (retention ratio, 0.44+/-0.08) when CPCs were delivered within the matrix, compared with cells injected alone (0.22+/-0.13, P=0.040) and with (18)F-FDG injected with or without the matrix (0.10+/-0.05 and 0.11+/-0.05, respectively, P<0.005). Tissue radionuclide biodistribution was performed after completion of positron-emission tomography imaging. When (18)F-FDG-labeled cells were injected with the collagen matrix, accumulation was significantly increased (by 69.6%, P=0.021) in the target ischemic hind limb muscle and significantly reduced (by 14.8% to 31.4%, P<0.05) in nonspecific tissues, compared with cells injected alone. Histology confirmed the increased retention in target tissue associated with the matrix. CONCLUSIONS: Early posttransplantation, a collagen matrix enhances progenitor cell retention and limits distribution to nonspecific tissues, as measured by the use of (18)F-FDG labeled cells and positron-emission tomography imaging and confirmed by biodistribution and histology. PMID- 19808544 TI - Evaluation of coronary atherosclerosis by multislice computed tomography in patients with acute myocardial infarction and without significant coronary artery stenosis: a comparative study with quantitative coronary angiography. AB - BACKGROUND: It is known that 9% to 31% of women and 4% to 14% of men with acute myocardial infarction have normal coronary arteries or nonsignificant coronary disease at angiography. These patients represent a diagnostic and therapeutic challenge. Multislice computed tomography (CT) can noninvasively identify the presence of coronary plaques even in the absence of significant coronary artery stenosis. This study evaluated the role of 64-slice CT, in comparison with coronary angiography, in detecting and characterizing coronary atherosclerosis in patients with acute myocardial infarction without significant coronary artery stenosis. METHODS AND RESULTS: Thirty consecutive patients with acute myocardial infarction but without significant coronary stenosis at coronary angiography underwent 64-slice CT. All coronary segments were quantitatively analyzed by means of coronary angiography (CA-QCA) and 64-slice CT (CT-QCA). Forty-seven (10.4%) of the 450 coronary segments were not evaluable by CT. The mean proximal reference diameters at CT-QCA and CA-QCA were, respectively, 2.88+/-0.75 mm and 2.65+/-0.9 mm; the overall correlation between CT-QCA and CA-QCA for quantification of reference diameter was r(s)=0.77; P<0.001. The mean percent stenosis was 14.4+/-8.0% at CT-QCA and 4.0+/-11.0% at CA-QCA and the correlation was r(s)=0.11; P=0.03. Overall CT-QCA showed the presence of 50 plaques, of which only 11 were detected by CA-QCA. CT-QCA identified 25 plaques in infarct-related coronary arteries. Positive remodeling was present in 38 of the 50 plaques (76%), with a higher prevalence in the coronary plaques not visualized by CA-QCA (82.1% versus 54.5%). CONCLUSIONS: CT-QCA correlates well with CA-QCA in terms of coronary reference diameter analysis, but not stenosis quantification. Multislice CT can detect coronary atherosclerotic plaques in segments of nonstenotic coronary arteries that are underestimated by CA and may have an incremental diagnostic value for the diagnosis of acute myocardial infarction in patients without significant coronary stenosis at CA. PMID- 19808545 TI - Development of receptor for advanced glycation end products-directed imaging of atherosclerotic plaque in a murine model of spontaneous atherosclerosis. AB - BACKGROUND: The receptor for advanced glycation end products (RAGE) is implicated in the development and progression of atherosclerosis. We tested the hypothesis that (99m)Tc-labeled anti-RAGE F(ab')(2) can be used as a noninvasive tool to image atherosclerotic lesions in apolipoprotein E-deficient (apoE(-/-)) mice. METHODS AND RESULTS: A sequence in the V-type Ig extracellular domain of RAGE was used to develop a peptide injected into rabbits; serum was retrieved, IgG prepared and affinity-purified, and pepsin-digested into F(ab')(2). Thirteen 6 week apoE(-/-) mice were fed a Western diet. At 20 weeks, 6 were injected with 15.2+/-1.9 MBq (350 to 411 microCi) (99m)Tc-labeled anti-RAGE F(ab')(2), 6 were injected with (99m)Tc-labeled control nonspecific IgG F(ab')(2), and 1 was injected with dual-labeled (99m)Tc and rhodamine anti-RAGE F(ab')(2). Four 20 week C57BL/6 mice were injected with (99m)Tc-labeled anti-RAGE F(ab')(2). All mice were imaged on a high resolution mini-gamma camera 4 hours after injection and euthanized. The aortic tree was dissected and photographed, and the proximal aorta was sectioned for staining after gamma scintillation counting. All 6 apoE( /-) mice injected with (99m)Tc-labeled anti-RAGE F(ab')(2) fragments showed focal tracer uptake in the proximal aorta (mean %ID/g, 1.98%). Disease and antibody controls showed no focal tracer uptake in the thorax (%ID/g, <1.0%). Histological sections of the proximal aorta showed American Heart Association class III lesions with lipid laden macrophages, smooth muscle cells, and positive staining for RAGE. On immunofluorescence, RAGE colocalized with macrophages. CONCLUSIONS: These data show the feasibility of noninvasively imaging RAGE in atherosclerotic lesions in a murine model and confirm levels of RAGE expression sufficient to allow detection on in vivo imaging. PMID- 19808546 TI - In vivo serial assessment of aortic aneurysm formation in apolipoprotein E deficient mice via MRI. AB - BACKGROUND: Hyperlipidimic mice administered angiotensin II have been used for the study of abdominal aortic aneurysms (AAAs). The purpose of this study was to examine the use of MRI for studying AAA development and for examining the effects of pharmacological intervention on AAA development in the apolipoprotein E deficient mouse. METHODS AND RESULTS: Suprarenal aortic aneurysms were generated in apolipoprotein E-deficient mice administered angiotensin II (1000 ng/kg per min) for up to 28 days. In vivo MRI was performed serially (once weekly) to assess AAA development and rupture. Comparison of AAA size as measured by in vivo and ex vivo MRI resulted in excellent agreement (r=0.96, P<0.0001). In addition, MRI correlated with histology-derived AAA area assessment (in vivo versus histology: r=0.84, P<0.0001; ex vivo versus histology: r=0.89, P<0.0001). In a separate study, angiotensin II-administered apolipoprotein E-deficient mice were treated with doxycycline (broad-based matrix metalloproteinase inhibitor; 30 mg/kg per day for 28 days). MRI was able to noninvasively assess a reduced rate of AAA development (46% versus 71%, P<0.05), a decreased AAA area (2.56 versus 4.02 mm(2), P<0.01), and decreased incidence of rupture (43% versus 100%) in treated versus control animals. Inhibition of aorta matrix metalloproteinase 2/9 activity was observed in the treated animals. CONCLUSIONS: These results demonstrate the use of MRI to noninvasively and temporally assess AAA development on pharmacological intervention in this preclinical cardiovascular disease model. PMID- 19808547 TI - Myocardial alterations in senescent mice and effect of exercise training: a strain rate imaging study. AB - BACKGROUND: Aging is accompanied by an alteration in myocardial contractility. However, its noninvasive detection is difficult. The effect of chronic exercise on this decrease is unknown. Murine models of senescence are increasingly used to test therapies in aging. We tested whether strain rate imaging detected left ventricular (LV) systolic dysfunction in aging mice and was able to assess a potential improvement after exercise. METHODS AND RESULTS: Young (3 weeks), adult (2 to 3 months), and old (6 to 18 months) C57BL6 male mice underwent echocardiograms with strain rate imaging, either in sedentary conditions or before, 2 weeks and 4 weeks after chronic swimming. Hemodynamic parameters of LV function including maximal and end-systolic elastance were obtained before euthanizing. LV fibrosis was measured using Sirius red staining. Conventional echocardiography was unable to detect LV systolic dysfunction in old mice, whereas both systolic strain rate and load-independent hemodynamic parameters such as preload recruitable stroke work and end-systolic elastance were significantly decreased. Both strain rate and load-independent hemodynamic parameters normalized after 4 weeks of exercise. Both endocardial and epicardial fibrosis were increased in the LV of aging mice. Endocardial fibrosis decreased in exercised aged mice. CONCLUSIONS: Strain rate noninvasively detects LV systolic dysfunction associated with aging in mice, whereas conventional echocardiography does not. Chronic exercise normalizes LV systolic function and decreases fibrosis in old mice. Strain rate imaging in mice may be a useful tool to monitor the effect of new therapeutic strategies preventing the myocardial dysfunction associated with aging. PMID- 19808548 TI - Intrinsic gating for small-animal computed tomography: a robust ECG-less paradigm for deriving cardiac phase information and functional imaging. AB - BACKGROUND: A projection-based method of intrinsic cardiac gating in small-animal computed tomography imaging is presented. METHODS AND RESULTS: In this method, which operates without external ECG monitoring, the gating reference signal is derived from the raw data of the computed tomography projections. After filtering, the derived gating reference signal is used to rearrange the projection images retrospectively into data sets representing different time points in the cardiac cycle during expiration. These time-stamped projection images are then used for tomographic reconstruction of different phases of the cardiac cycle. Intrinsic gating was evaluated in mice and rats and compared with extrinsic retrospective gating. An excellent agreement was achieved between ECG derived gating signal and self-gating signal (coverage probability for a difference between the 2 measurements to be less than 5 ms was 89.2% in mice and 85.9% in rats). Functional parameters (ventricular volumes and ejection fraction) obtained from the intrinsic and the extrinsic data sets were not significantly different. The ease of use and reliability of intrinsic gating were demonstrated via a chemical stress test on 2 mice, in which the system performed flawlessly despite an increased heart rate. Because of intrinsic gating, the image quality was improved to the extent that even the coronary arteries of mice could be visualized in vivo despite a heart rate approaching 430 bpm. Feasibility of intrinsic gating for functional imaging and assessment of cardiac wall motion abnormalities was successfully tested in a mouse model of myocardial infarction. CONCLUSIONS: Our results demonstrate that self-gating using advanced software postprocessing of projection data promises to be a valuable tool for rodent computed tomography imaging and renders ECG gating with external electrodes superfluous. PMID- 19808549 TI - Multimodality cardiovascular molecular imaging, part I. AB - In Part I of this consensus article, the imaging methodology, evolving imaging technology, and development of novel targeted molecular probes relevant to the developing field of cardiovascular molecular imaging were reviewed. Novel reporter gene and reporter probe imaging approaches for tracking of cardiac transgene expression were also discussed and have important future implications for evaluation of gene- and cell-based therapies for the failing heart. The current role of metabolic and receptor imaging was also briefly reviewed, as these represent the beginning of our clinical application of molecular imaging within the cardiovascular system. Part II will summarize the available targeted imaging probes as well as specific future applications of molecular imaging for identification and evaluation of critical pathophysiological processes of the cardiovascular system. PMID- 19808550 TI - Noninvasive diagnosis and prognosis assessment in chronic coronary artery disease: stress testing with and without imaging perspective. PMID- 19808551 TI - Noninvasive diagnostic and prognostic assessment of individuals with suspected coronary artery disease: coronary computed tomographic angiography perspective. PMID- 19808552 TI - Three-dimensional transesophageal echocardiography in a patient with early failure of mitral valve repair: why are we still looking at a three-dimensional structure in 2 dimensions? PMID- 19808554 TI - Sequential development of multiple mechanical complications of myocardial infarction. PMID- 19808555 TI - Periannular abscess cavities in endocarditis: the case for prolonged surveillance. PMID- 19808556 TI - CT angiography: first things first. PMID- 19808557 TI - Insights into myocardial microstructure during infarct healing and remodeling: pathologists need not apply. PMID- 19808558 TI - The role of left atrial function in diastolic heart failure. PMID- 19808559 TI - Left atrial function in diastolic heart failure. AB - BACKGROUND: Abnormalities in left ventricular (LV) structure and function occur in patients with diastolic heart failure (DHF). The reasons for the transition from asymptomatic dysfunction to heart failure need better definition, including noninvasive measurements that can detect the transition. METHODS AND RESULTS: In 64 patients undergoing right heart catheterization, simultaneous echocardiographic imaging was performed. As a control group, 27 healthy subjects were included. There were 25 with ejection factor (EF) <50%, 20 in DHF, and 19 with normal EF and LV hypertrophy but not in heart failure (diastolic dysfunction). LV volumes, mass, left atrial (LA) volumes and EF, annular atrial velocity (a'), and LA strain during systole (LA(S)), and atrial contraction (LA(A)) were measured. The ratio of wedge pressure to LA(S) strain was used as an index of LA stiffness, as was the ratio of E/e' to LA(S) strain. All 3 patient groups had increased LA volumes and depressed LA EF, a', and LA(A) strain, with no significant difference between patients with DHF and diastolic dysfunction in LA systolic function indices, LV mass, LA volumes, LV, and arterial elastance. LA(S) strain was lower in patients with DHF, and LA stiffness (invasive and noninvasive) was higher (both P<0.01), related well to pulmonary artery systolic pressure (r=0.79, P<0.001), and was most accurate in identifying DHF patients from those with diastolic dysfunction (invasive area under the curve: 0.93, noninvasive: 0.85). CONCLUSIONS: Patients with DHF have increased LV mass and LA volume in comparison with normal controls, but not versus patients with LV hypertrophy who are not in heart failure. On the other hand, LA(S) strain is significantly reduced and LA stiffness is the most accurate index in identifying patients with DHF. PMID- 19808560 TI - Diagnostic accuracy and impact of computed tomographic coronary angiography on utilization of invasive coronary angiography. AB - BACKGROUND: Computed tomographic coronary angiography (CTA), given its high negative predictive value, is a potential gatekeeper for invasive coronary angiography (ICA). Before CTA can be further accepted into clinical practice, its impact on healthcare resources needs to be better understood. We sought to determine the clinical impact of CTA on ICA referrals, CTA accuracy, and normalcy rate. METHODS AND RESULTS: To determine the impact of CTA, consecutive patients (n=7017) undergoing ICA before and after implementing a dedicated cardiac CT program were reviewed and compared with 3 other centers (n=11 508). To determine CTA accuracy, we evaluated consecutive CTA patients who underwent ICA. For normalcy rate, we identified patients with a low pretest probability for obstructive coronary artery disease. With the implementation of a cardiac CT program, the frequency of normal ICA decreased from 31.5% (1114 of 3538 patients) to 26.8% (932 of 3479 patients) (P<0.001). These findings were significantly different (P=0.003) from the 3 centers, in which normal ICAs were unchanged (30.0% [1870 of 6224 patients] to 31.0% [1642 of 5284 patients]). CTA had excellent per-patient sensitivity (99% [CI, 95% to 100%]), positive predictive value (92% [CI, 86% to 96%]) and negative predictive value (95% [CI, 72% to 100%]). Because of referral bias, specificity (64% [CI, 44% to 81%]) was low; however, the normalcy rate of CTA was 94% (CI, 90% to 97%). After adjusting for referral bias, the adjusted sensitivity was 90% (CI, 89% to 91%), and the adjusted specificity was 95% (CI, 94% to 96%), with positive and negative predictive values of 92% (CI, 91% to 93%) and 93% (CI, 92% to 94%), respectively. CONCLUSIONS: The clinical implementation of CTA appears to positively impact ICA by reducing the frequency of normal ICA. The operating characteristics of CTA support its potential role as a tool useful in ruling out obstructive coronary artery disease. PMID- 19808561 TI - A study of functional anatomy of aortic-mitral valve coupling using 3D matrix transesophageal echocardiography. AB - BACKGROUND: Mitral and aortic valves are known to be coupled via fibrous tissue connecting the two annuli. Previous studies evaluating this coupling have been limited to experimental animals using invasive techniques. The new matrix array transesophageal transducer provides high-resolution real-time 3D images of both valves simultaneously. We sought to develop and test a technique for quantitative assessment of mitral and aortic valve dynamics and coupling. METHODS AND RESULTS: Matrix array transesophageal (Philips iE33) imaging was performed in 24 patients with normal valves who underwent clinically indicated transesophageal echocardiography. Custom software was used to detect and track the mitral and aortic annuli in 3D space throughout the cardiac cycle, allowing automated measurement of changes in mitral and aortic valve morphology. Mitral annulus surface area and aortic annulus projected area changed reciprocally over time. Mitral annulus surface area was 8.0+/-2.1 cm(2) at end-diastole and decreased to 7.7+/-2.1 cm(2) in systole, reaching its maximum (10.0+/-2.2 cm(2)) at mitral valve opening. Aortic annulus projected area was 4.1+/-1.2 cm(2) at end-diastole, then increased during isovolumic contraction reaching its maximum (4.8+/-1.3 cm(2)) in the first third of systole and its minimum (3.6+/-1.0 cm(2)) during isovolumic relaxation. The angle between the mitral and aortic annuli was maximum (136+/-13 degrees ) at end-diastole and decreased to its minimum value (129+/-11 degrees ) during systole. CONCLUSIONS: This is the first study to report quantitative 3D assessment of the mitral and aortic valve dynamics from matrix array transesophageal images and describe the mitral-aortic coupling in a beating human heart. This ability may have impact on patient evaluation for valvular surgical interventions and prosthesis design. PMID- 19808562 TI - Sequential changes of myocardial microstructure in patients postmyocardial infarction by diffusion-tensor cardiac MR: correlation with left ventricular structure and function. AB - BACKGROUND: We used diffusion-tensor cardiac MR to investigate myocardial microstructure changes, including tissue integrity (mean diffusivity [MD], fractional anisotropy) and fiber architecture (helix angles) in patients with recent myocardial infarction (MI). This study aimed to investigate the sequential changes of myocardial microstructure and its relationships with changes of macrostructure and function of the left ventricle post-MI. METHODS AND RESULTS: Seventeen patients (age, 55.1+/-11.5 years; all men) participated in the follow up study. Diffusion-tensor cardiac MR, cine gradient echo for left ventricle function, and late gadolinium enhancement for viability were measured from recent to chronic MI (median interval, 191 days). When compared with the remote zone, the infarct-adjacent zone showed overall increase of MD (2-way MANOVA, F(1,16)=36.3; P<0.001), decrease of fractional anisotropy (F(1,16)=5.8; P=0.029), and decrease of mean helix angles (F(1,16)=62.0; P<0.001). From recent to chronic MI, there was overall sequential decrease of MD (F(1,16)=22.6; P<0.001) and increase of fractional anisotropy (F(1,16)=7.8; P=0.013). Multiple linear regression showed that the improvement of wall thickening in the infarct-adjacent zone correlated with sequential decrease of MD in the infarct-adjacent zone (r= 0.70; P=0.002) and increase of mean helix angles (ie, more right-handed helical myofiber reorientation, predominantly subendocardial location) in the remote zone (r=0.60; P=0.011). Likewise, wall thickening in the remote zone correlated with MD in the remote zone (r=-0.72; P=0.001) and mean helix angles in the infarct adjacent zone (r=0.72; P=0.001). CONCLUSIONS: Diffusion-tensor cardiac MR suggests that sequential zonal improvement of tissue integrity and fiber architecture remodeling both associate with sequential recovery of zonal wall thickening of the left ventricle from recent to chronic MI. PMID- 19808563 TI - Age- and gender-specific changes in the left ventricular relaxation: a Doppler echocardiographic study in healthy individuals. AB - BACKGROUND: Although left ventricular diastolic function has been shown to deteriorate with advancing age, its gender-specific change is unknown. The aim of this study was to investigate age- and gender-specific changes in tissue Doppler derived left ventricular diastolic index, E'. METHODS AND RESULTS: A total of 1333 healthy individual without known heart disease or hypertension (mean age, 55 years; range, 10 to 89) were enrolled and studied. Peak early mitral annular velocity (E') and peak late mitral annular velocity (A') were recorded and measured. As an index of the left ventricular relaxation, E' was used. As an index of the left ventricular filling pressure, E/E' was calculated. Although systolic indices poorly correlated with age, diastolic indices correlated well with age. Among those aged 30 to 39 and 40 to 49 years, E' was significantly lower in males than in females. In subjects aged 50 to 59 and 60 to 69 years, E' was similar in both genders. Among those aged 70 to 79 and 80 to 89 years, E' was significantly lower in females than in males. Predictors of the lowest quartile of E' among subjects aged >50 years were age (P<0.0001; chi(2)=66.11; odds ratio, 1.08; 95% CI, 1.058 to 1.097) and female gender (P=0.002; chi(2)=9.23; odds ratio, 1.68; 95% CI, 1.202 to 2.343). CONCLUSIONS: Age-related changes in diastolic indices were gender specific. In the elderly population, diastolic function deteriorated more significantly in the female gender than in the male gender. These results may explain the relatively higher incidence in elderly females among patients with diastolic heart failure and higher cardiovascular mortality in the female gender. PMID- 19808564 TI - Rapid initial reduction of hyperenhanced myocardium after reperfused first myocardial infarction suggests recovery of the peri-infarction zone: one-year follow-up by MRI. AB - BACKGROUND: The time course and magnitude of infarct involution, functional recovery, and normalization of infarct-related electrocardiographic (ECG) changes after acute myocardial infarction (MI) are not completely known in humans. We sought to explore these processes early after MI and during infarct-healing using cardiac MRI. METHODS AND RESULTS: Twenty-two patients with reperfused first-time MI were examined by MRI and ECG at 1, 7, 42, 182, and 365 days after infarction. Global left ventricular function and regional wall thickening were assessed by cine MRI, and injured myocardium was depicted by delayed contrast-enhanced MRI. Infarct size by ECG was estimated by QRS scoring. The reduction of hyperenhanced myocardium occurred predominantly during the first week after infarction (64% of the 1-year reduction). Furthermore, during the first week the amount of nonhyperenhanced myocardium increased significantly (P<0.001), although the left ventricular mass remained unchanged. Left ventricular ejection fraction increased gradually, whereas the greater the regional transmural extent of hyperenhancement at day 1, the later the recovery of regional wall thickening. Regional wall thickening decreased progressively with increasing initial transmural extent of hyperenhancement (P(trend)<0.0001). The time course and magnitude of decrease in QRS score corresponded with the reduction of hyperenhanced myocardium. CONCLUSIONS: The early reduction of hyperenhanced myocardium may reflect recovery of hyperenhanced, reversibly injured myocardium, which must be considered when predicting functional recovery from delayed contrast-enhanced MRI findings early after infarction. Also, the time course and magnitude for reduction of hyperenhanced myocardium were associated with normalization of infarct-related ECG changes. PMID- 19808566 TI - Endocarditis after pectus excavatum repair: a case report. PMID- 19808567 TI - Multifocal left ventricular pseudoaneurysm 25 years after aneurysm repair: detection by 64-detector computed tomographic coronary angiography. PMID- 19808565 TI - Multimodality cardiovascular molecular imaging, Part II. PMID- 19808568 TI - Successful deployment of a transcatheter aortic valve in bicuspid aortic stenosis: role of imaging with multislice computed tomography. PMID- 19808569 TI - Right aortic arch related to Kommerell diverticulum and internal carotid artery agenesis. PMID- 19808570 TI - A histological "fly-through" of a diseased coronary artery. PMID- 19808571 TI - A novel (18)F-labeled tracer for the quantification of myocardial blood flow and infarct size with positron-emission tomography: another way to avoid the need of an on-site cyclotron. PMID- 19808572 TI - Evaluation of a novel (18)F-labeled positron-emission tomography perfusion tracer for the assessment of myocardial infarct size in rats. AB - BACKGROUND: The goal of this study was to evaluate a new (18)F-labeled positron emission tomography (PET) perfusion tracer, (18)F BMS747158-02, for the assessment of myocardial infarct (MI) size. METHODS AND RESULTS: Wistar rats were studied 24 hours after ligation of the left coronary artery either permanently (n=15) or transiently (n=16) for 30 minutes. Seven nonoperated rats were studied as controls. The rats were injected with 37 MBq of (18)F BMS747158-02 and imaged with a small animal PET scanner for 20 minutes. Polar maps were generated for measurement of PET defect size, and left ventricular systolic and diastolic volumes were assessed in gated images. As a reference, MI size was determined by 2,3,5-triphenyltetrazolium chloride staining of left ventricular tissue samples. Permanent or transient ligation of the left coronary artery produced transmural or subendocardial MI of variable sizes, respectively. In normal rats, PET imaging demonstrated intense and homogeneous uptake of (18)F BMS747158-02 throughout the myocardium. After ligation, sharply defined perfusion defects were present. Throughout the imaging period, the defect size correlated closely with the MI size either after permanent (r=0.88; P<0.01; mean difference, 1.86%) or transient (r=0.92; P<0.01; mean difference, 2.16%) ligation of the left coronary artery. Moreover, reduction of left ventricular systolic function measured with PET correlated with the MI size (r=-0.81; P<0.01; n=23). CONCLUSIONS: Myocardial (18)F BMS747158-02 PET imaging provides excellent image quality and uptake properties, enabling accurate evaluation of MI size and left ventricular function in rats. It is a promising technique for evaluation of MI size in clinical trials. PMID- 19808573 TI - Reduced systolic torsion in chronic "pure" mitral regurgitation. AB - BACKGROUND: Global left ventricular (LV) torsion declines with chronic ischemic mitral regurgitation (MR), which may accelerate the LV remodeling spiral toward global cardiomyopathy; however, it has not been definitively established whether this torsional decline is attributable to the infarct, the MR, or their combined effect. We tested the hypothesis that chronic "pure" MR alone reduces global LV torsion. METHODS AND RESULTS: Chronic "pure" MR was created in 13 sheep by surgically punching a 3.5- to 4.8-mm hole (HOLE) in the mitral valve posterior leaflet. Nine control (CNTL) sheep were operated on concurrently. At 1 (WK-01) and 12 weeks (WK-12) postoperatively, the 4D motion of implanted radiopaque markers was used to calculate global LV torsion. MR-grade in HOLE was greater than CNTL at WK-01 and WK-12 (2.5+/-1.1 versus 0.6+/-0.5, P<0.001 at WK-12). HOLE LV mass index was larger at WK-12 compared with CNTL (195+/-14 versus 170+/-17 g/m(2), P<0.01), indicating LV remodeling. Global LV systolic torsion decreased in HOLE from WK-01 to WK-12 (4.1+/-2.8 degrees versus 1.7+/-1.7 degrees , P<0.01), but did not change in CNTL (5.5+/-1.8 degrees versus 4.2+/-2.7 degrees , P=NS). Global LV torsion was lower in HOLE relative to CNTL at WK-12 (P<0.05) but not at WK-01 (P=NS). CONCLUSIONS: Twelve weeks of chronic "pure" MR resulting in mild global LV remodeling is associated with significantly increased LV mass index and reduced global LV systolic torsion, but no other significant changes in hemodynamics. MR alone is a major component of torsional deterioration in "pure" MR and may be an important factor in chronic ischemic mitral regurgitation. PMID- 19808574 TI - Impact of mild hypertension on left atrial size and function. AB - BACKGROUND: Left atrial (LA) enlargement has been documented to occur in moderate and severe hypertension. METHODS AND RESULTS: One hundred twelve mild hypertension patients were prospectively recruited and compared with 198 healthy volunteers. All recruits had a transthoracic echocardiogram. Maximum LA biplane volume, minimum LA biplane volume, and pre 'p'-LA biplane volume were measured, and left atrial passive, active emptying, and conduit volumes were calculated at baseline and in a subgroup of patients after 12 months. After adjusting for age, gender, and body mass index, maximum LA biplane volume, pre 'p'-LA biplane volume, and their indexed volumes were increased in the hypertension group. Active emptying volume and fraction were significantly increased in the hypertension group, with no change in conduit and passive volumes. Subgroup analysis comparing hypertensives with normal/mildly increased left ventricular mass (group 1) with those with moderate/severely increased left ventricular mass (group 2) at baseline demonstrated that maximum LA biplane volume (62.8+/-17.9 mL versus 45.4+/-13.7 mL; P<0.001) was significantly increased in group 2. Active emptying volume was also increased. CONCLUSIONS: Even mild hypertension seems to be associated with a reduction in early diastolic filling. This results in augmented late left ventricular diastolic filling due to active atrial contraction and may be the mechanism for the increase in left atrial size. PMID- 19808575 TI - Prevalence of coronary artery disease assessed by multislice computed tomography coronary angiography in patients with paroxysmal or persistent atrial fibrillation. AB - BACKGROUND: Although atrial fibrillation (AF) has been linked to underlying coronary artery disease (CAD), data supporting this association have been based on ECG and clinical history for the definition of CAD rather than direct visualization of atherosclerosis. METHODS AND RESULTS: The prevalence of CAD among patients with paroxysmal or persistent AF and without history of CAD was evaluated using multislice computed tomography. Multislice computed tomography was performed in 150 patients with AF (61+/-11 years, 67% males, 58% asymptomatic) with predominantly low (59%) or intermediate (25%) pretest likelihood of CAD. CAD was classified as obstructive (> or =50% luminal narrowing) or not. A population of 148 patients without history of AF, similar to the AF group as to age, gender, symptomatic status, and pretest likelihood, served as a control group. Logistic regression analysis was applied to evaluate the relationship between demographic and clinical data and the presence of obstructive CAD. On the basis of multislice computed tomography, 18% of patients with AF were classified as having no CAD, whereas 41% showed nonobstructive CAD and the remaining 41% had obstructive CAD. Among patients without AF, 32% were classified as having no CAD, whereas 41% showed nonobstructive CAD and 27% had obstructive CAD (P=0.010 compared with patients with AF). At logistic regression analysis, age, male gender, and the presence of AF were significantly related to obstructive CAD. CONCLUSIONS: A higher prevalence of obstructive CAD was observed among patients with AF, confirming the hypothesis that AF could be a marker of advanced coronary atherosclerosis. PMID- 19808576 TI - Relationships among regional arterial inflammation, calcification, risk factors, and biomarkers: a prospective fluorodeoxyglucose positron-emission tomography/computed tomography imaging study. AB - BACKGROUND: Fluorodeoxyglucose positron-emission tomography (FDG PET) imaging of atherosclerosis has been used to quantify plaque inflammation and to measure the effect of plaque-stabilizing drugs. We explored how atherosclerotic plaque inflammation varies across arterial territories and how it relates to arterial calcification. We also tested the hypotheses that the degree of local arterial inflammation measured by PET is correlated with the extent of systemic inflammation and presence of risk factors for vascular disease. METHODS AND RESULTS: Forty-one subjects underwent vascular PET/computed tomography imaging with FDG. All had either vascular disease or multiple risk factors. Forty subjects underwent carotid imaging, 27 subjects underwent aortic, 24 subjects iliac, and 13 subjects femoral imaging. Thirty-three subjects had a panel of biomarkers analyzed. We found strong associations between FDG uptake in neighboring arteries (left versus right carotid, r=0.91, P<0.001; ascending aorta versus aortic arch, r=0.88, P<0.001). Calcification and inflammation rarely overlapped within arteries (carotid artery FDG uptake versus calcium score, r= 0.42, P=0.03). Carotid artery FDG uptake was greater in those with a history of coronary artery disease (target-to- BACKGROUND: <0.01) and in males versus females (target-to- BACKGROUND: <0.05). Similar findings were also noted in the aorta and iliac arteries. Subjects with the highest levels of FDG uptake also had the greatest concentrations of inflammatory biomarkers (descending aorta target to- BACKGROUND: =0.53, P=0.01; carotid target-to- BACKGROUND: =0.50, P=0.01). Nonsignificant positive trends were seen between FDG uptake and levels of interleukin-18, fibrinogen, and C-reactive protein. Finally, we found that the atheroprotective biomarker adiponectin was negatively correlated with the degree of arterial inflammation in the descending aorta (r=-0.49, P=0.03). CONCLUSIONS: This study shows that FDG PET imaging can increase our knowledge of how atherosclerotic plaque inflammation relates to calcification, serum biomarkers, and vascular risk factors. Plaque inflammation and calcification rarely overlap, supporting the theory that calcification represents a late, burnt-out stage of atherosclerosis. Inflammation in one arterial territory is associated with inflammation elsewhere, and the degree of local arterial inflammation is reflected in the blood levels of several circulating biomarkers. We suggest that FDG PET imaging could be used as a surrogate marker of both atherosclerotic disease activity and drug effectiveness. Prospective, event-driven studies are now underway to determine the role of this technique in clinical risk prediction. PMID- 19808577 TI - Strain-encoded MRI for evaluation of left ventricular function and transmurality in acute myocardial infarction. AB - BACKGROUND: Strain-encoded imaging (SENC) is a new technique for myocardial deformation analysis in cardiac MRI. The aim of the study was, therefore, to evaluate whether myocardial deformation imaging performed by SENC allows for quantification of regional left ventricular function and is related to transmurality states of infarcted tissue in patients with acute myocardial infarction. METHODS AND RESULTS: Cardiac MRI was performed in 38 patients with acute myocardial infarction 3+/-1 days after successful reperfusion using a clinical 1.5-T MRI scanner. Ten healthy volunteers served as controls. SENC is a technique that directly measures peak circumferential strain from long-axis views and peak longitudinal strain from short-axis views. Measurements were obtained for each segment in a modified 17-segment model. Wall motion and infarcted tissue were evaluated semiquantitatively from steady-state free-precession cine sequences and contrast-enhanced MR images and were then related to myocardial strain. Comparison of peak circumferential strain assessed by SENC and MR tagging was performed. In total, 456 segments were analyzed. Peak circumferential and longitudinal strain calculated from SENC images was significantly different in regions defined as normokinetic, hypokinetic, or akinetic (P<0.001). A cutoff peak systolic circumferential strain value of -10% differentiated nontransmural from transmural infarcted myocardium, with a sensitivity of 97% and a specificity of 94%. Strain analysis of SENC and MR tagging correlated well (r=0.76) with narrow limits of agreement (-9.9% to 8.5%). CONCLUSIONS: SENC provides rapid and objective quantification of regional myocardial function and allows discrimination between different transmurality states in patients with acute myocardial infarction. PMID- 19808578 TI - Apical rotation assessed by speckle-tracking echocardiography as an index of global left ventricular contractility. AB - BACKGROUND: Left ventricular (LV) apical rotation and twist can be estimated noninvasively by speckle-tracking echocardiography (STE). In this study, we tested whether apical rotation is an accurate index of LV contractility. METHODS AND RESULTS: We measured LV basal and apical rotation by STE in 11 open-chest anesthetized mongrel dogs under 8 different inotropic stages before and after ligation of either left anterior descending (n=6) or circumflex coronary artery (n=5). We measured LV pressure simultaneously with a high-fidelity pressure catheter and calculated LV ejection fraction (EF) with the biplane Simpson method and 2D echocardiography. Maximal positive dP/dt (dP/dt(max)) was used as the gold standard measurement of LV contractility. We compared LV twist and apical rotation and EF against dP/dt(max) by linear mixed model. LV apical rotation and twist showed dose-dependent increases and decreases after dobutamine and esmolol infusion, respectively. However, basal rotation did not change significantly during different inotropic conditions. There was a stronger association between dP/dt(max) and LV twist (R(2)=0.747, P<0.001) and apical rotation (R(2)=0.726, P<0.001) than between dP/dt(max) and EF (R(2)=0.408, P<0.001), and this trend was more apparent with coronary ligation irrespective of the ligation site. There was also a high association between dP/dt(max) and apical rotation alone, both with (R(2)=0.805, P<0.001) and without (R(2)=0.748, P<0.001) coronary ligation. Apical rotation alone showed comparable accuracy to LV twist. Apical rotational velocity also showed a high association with dP/dt(max) (R(2)=0.669, P<0.001) and LV twist (R(2)=0.892, P<0.001). CONCLUSIONS: Apical rotation assessed by STE is an effective noninvasive index of global LV contractility and is more closely related to dP/dt(max) than LV EF. PMID- 19808579 TI - Strain-encoded cardiac MRI as an adjunct for dobutamine stress testing: incremental value to conventional wall motion analysis. AB - BACKGROUND: High-dose dobutamine stress MRI is safe and feasible for the diagnosis of coronary artery disease (CAD) in humans. However, the assessment of cine scans relies on the visual interpretation of regional wall motion, which is subjective. Recently, strain-encoded MRI (SENC) has been proposed for the direct color-coded visualization of myocardial strain. The purpose of our study was to compare the diagnostic value of SENC with that provided by conventional wall motion analysis for the detection of inducible ischemia during dobutamine stress MRI. METHODS AND RESULTS: Stress-induced ischemia was assessed by wall motion analysis and by SENC in 101 patients with suspected or known CAD and in 17 healthy volunteers who underwent dobutamine stress MRI in a clinical 1.5-T scanner. Quantitative coronary angiography deemed as the standard reference for the presence or absence of significant CAD (> or =50% diameter stenosis). On a coronary vessel level, SENC detected inducible ischemia in 86 of 101 versus 71 of 101 diseased coronary vessels (P<0.01 versus cine) and showed normal strain response in 189 of 202 versus 194 of 202 vessels with <50% stenosis (P=NS versus cine). On a patient level, SENC detected inducible ischemia in 63 of 64 versus 55 of 64 patients with CAD (P<0.05 versus cine) and showed normal strain response in 32 of 37 versus 34 of 37 patients without CAD (P=NS versus cine). Quantification analysis demonstrated a significant correlation between strain rate reserve and coronary artery stenosis severity (r(2)=0.56, P<0.001), and a cutoff value of strain rate reserve of 1.64 was deemed as a highly accurate marker for the detection of > or =50% stenosis (area under the curve, 0.96; SE, 0.01; 95% CI, 0.94 to 0.98; P<0.001). CONCLUSIONS: The direct color-coded visualization of strain on MR images is a useful adjunct for dobutamine stress MRI, which provides incremental value for the detection of CAD compared with conventional wall motion readings on cine images. PMID- 19808580 TI - Echocardiographic assessment of percutaneous patent foramen ovale and atrial septal defect closure complications. PMID- 19808581 TI - Atherosclerosis imaging: prognostically useful or merely more of what we know? PMID- 19808582 TI - A giant superior venous cava resulted from obstructive total anomalous pulmonary venous connection. PMID- 19808583 TI - Cardiovascular images. Giant right coronary aneurysm to left ventricular fistula. PMID- 19808584 TI - Cardiovascular images. Detecting transient myocardial ischemia in the context of acute coronary syndrome in the emergency department: Delta map analysis of body electrocardiographic surface mapping. PMID- 19808585 TI - Cardiocascular images. M-mode echocardiographic demonstration of atrial flutter. PMID- 19808586 TI - Anatomic and functional assessment of coronary artery disease: convergence of 2 aims in a single setting. PMID- 19808587 TI - Coronary artery or myocyte: wherein lies the diagnosis? PMID- 19808588 TI - MRI of the microarchitecture of myocardial infarction: are we seeing new kinds of structures? PMID- 19808589 TI - Aortopulmonary collaterals in single-ventricle congenital heart disease: how much do they count? PMID- 19808591 TI - Infarct tissue heterogeneity assessed with contrast-enhanced MRI predicts spontaneous ventricular arrhythmia in patients with ischemic cardiomyopathy and implantable cardioverter-defibrillator. AB - BACKGROUND: The relation between infarct tissue heterogeneity on contrast enhanced MRI and the occurrence of spontaneous ventricular arrhythmia (or sudden cardiac death) is unknown. Therefore, the study purpose was to evaluate the predictive value of infarct tissue heterogeneity assessed with contrast-enhanced MRI on the occurrence of spontaneous ventricular arrhythmia with subsequent implantable cardioverter-defibrillator (ICD) therapy (as surrogate of sudden cardiac death) in patients with previous myocardial infarction. METHODS AND RESULTS: Ninety-one patients (age, 65+/-11 years) with previous myocardial infarction scheduled for ICD implantation underwent cine MRI to evaluate left ventricular function and volumes and contrast-enhanced MRI for characterization of scar tissue (infarct gray zone as measure of infarct tissue heterogeneity, infarct core, and total infarct size). Appropriate ICD therapy was documented in 18 patients (20%) during a median follow-up of 8.5 months (interquartile range, 2.1 to 20.3). Multivariable Cox proportional hazards analysis revealed that infarct gray zone was the strongest predictor of the occurrence of spontaneous ventricular arrhythmia with subsequent ICD therapy (hazard ratio, 1.49/10 g; CI, 1.01 to 2.20; chi(2)=4.0; P=0.04). CONCLUSIONS: Infarct tissue heterogeneity on contrast-enhanced MRI is the strongest predictor of spontaneous ventricular arrhythmia with subsequent ICD therapy (as surrogate of sudden cardiac death) among other clinical and MRI variables, that is, total infarct size and left ventricular function and volumes, in patients with previous myocardial infarction. PMID- 19808590 TI - Adenosine stress 64- and 256-row detector computed tomography angiography and perfusion imaging: a pilot study evaluating the transmural extent of perfusion abnormalities to predict atherosclerosis causing myocardial ischemia. AB - BACKGROUND: Multidetector computed tomography coronary angiography (CTA) is a robust method for the noninvasive diagnosis of coronary artery disease. However, in its current form, CTA is limited in its prediction of myocardial ischemia. The purpose of this study was to test whether adenosine stress computed tomography myocardial perfusion imaging (CTP), when added to CTA, can predict perfusion abnormalities caused by obstructive atherosclerosis. METHODS AND RESULTS: Forty patients with a history of abnormal single-photon emission computed tomography myocardial perfusion imaging (SPECT-MPI) underwent adenosine stress 64-row (n=24) or 256-row (n=16) detector CTP and CTA. A subset of 27 patients had invasive angiography available for quantitative coronary angiography. CTA and quantitative coronary angiography were evaluated for stenoses > or =50%, and SPECT-MPI was evaluated for fixed and reversible perfusion deficits using a 17-segment model. CTP images were analyzed for the transmural differences in perfusion using the transmural perfusion ratio (subendocardial attenuation density/subepicardial attenuation density). The sensitivity, specificity, positive predictive value, and negative predictive value for the combination of CTA and CTP to detect obstructive atherosclerosis causing perfusion abnormalities using the combination of quantitative coronary angiography and SPECT as the gold standard was 86%, 92%, 92%, and 85% in the per-patient analysis and 79%, 91%, 75%, and 92% in the per vessel/territory analysis, respectively. CONCLUSIONS: The combination of CTA and CTP can detect atherosclerosis causing perfusion abnormalities when compared with the combination of quantitative coronary angiography and SPECT. PMID- 19808592 TI - Age-related left ventricular remodeling and associated risk for cardiovascular outcomes: the Multi-Ethnic Study of Atherosclerosis. AB - BACKGROUND: Age-related alterations of left ventricular (LV) structure and function that may predispose to cardiovascular events are not well understood. METHODS AND RESULTS: We used cardiac MRI to examine age-related differences in LV structure and function in 5004 participants without overt cardiovascular disease when enrolled in the Multi-Ethnic Study of Atherosclerosis; 1099 participants received additional strain analyses by MRI tagging. We also assessed the relation of age-associated remodeling with cardiovascular outcomes using Cox proportional hazard models adjusting for cardiovascular risk factors. Although LV mass decreased with age (-0.3 g per year), the mass-to-volume ratio markedly increased (+5 mg/mL per year, P<0.0001), driven by a substantial reduction in end-diastolic volume (-0.8 mL per year, P<0.0001). Age was also associated with a significant fall in stroke volume (-0.4 mL per year, P<0.0001), along with strain patterns reflecting systolic (P<0.0001) as well as diastolic (P<0.01) myocardial dysfunction-despite a modestly enhanced ejection fraction (+0.1% per year, P<0.0001). Increased mass-to-volume ratio conferred a significant risk for total cardiovascular events; this trend was strongest among younger (<65 years; hazard ratio, 3.69 [CI, 1.34 to 10.10]) versus older (> or =65 years; hazard ratio, 1.68 [CI 0.77 to 3.68]) individuals with the highest compared to lowest mass-to-volume ratio quintile (P(interaction)=0.013). CONCLUSIONS: Age is associated with a phenotype of LV remodeling marked by increased mass-to-volume ratio and accompanied by systolic as well as diastolic myocardial dysfunction that is not reflected by preserved ejection fraction. This pattern of ventricular remodeling confers significant cardiovascular risk, particularly when present earlier in life. PMID- 19808593 TI - Sixty-four-slice multidetector computed tomography: an accurate imaging modality for the evaluation of coronary arteries in dilated cardiomyopathy of unknown etiology. AB - BACKGROUND: The goal of this study was to assess the safety, feasibility, and diagnostic accuracy of 64-slice multidetector computed tomography (MDCT) for the evaluation of coronary arteries in dilated cardiomyopathy (DCM) of unknown etiology. Sixteen-slice MDCT is useful in patients affected by DCM. However, technical limitations, such as cardiac arrhythmias, an inability of patients to sustain a long breath-hold, and the need of a high dose of contrast agent may limit its accuracy and widespread use. METHODS AND RESULTS: Invasive coronary angiography (ICA) and MDCT coronary angiography were performed on 132 consecutive patients (82 men; age 63+/-11 years) affected by DCM (ejection fraction, 34+/ 10%) of unknown etiology. In 2 patients (1.5%), MDCT was not feasible because of atrial fibrillation. Of the remaining 130 patients, 88 exhibited normal and 42 exhibited diseased coronary arteries in both MDCT and ICA. All patients with coronary artery disease except for 1 were correctly classified by MDCT as 1 vessel (11 cases), 2-vessel (13 cases), and 3-vessel (18 cases) disease. In the segment-based analyses, the overall feasibility for MDCT was 98.5% (1902 of 1930 segments). Segment-based and patient-based analyses for the detection of luminal stenosis of >50% and >70% were performed. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of MDCT for the detection of >50% stenosis were 98.1%, 99.9%, 98.7%, 99.8%, and 99.7%, respectively. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of MDCT for the detection of >70% stenosis were 99.5%, 98.6%, 94.1%, 99.9%, and 99.4%, respectively. CONCLUSIONS: Excellent feasibility and diagnostic accuracy, combined with low invasiveness, make 64 slice MDCT an ideal imaging modality for the anatomic evaluation of coronary circulation in patients with DCM of unknown etiology. PMID- 19808594 TI - Diffusion spectrum MRI tractography reveals the presence of a complex network of residual myofibers in infarcted myocardium. AB - BACKGROUND: Changes in myocardial microstructure are important components of the tissue response to infarction but are difficult to resolve with current imaging techniques. A novel technique, diffusion spectrum MRI tractography (DSI tractography), was thus used to image myofiber architecture in normal and infarcted myocardium. Unlike diffusion tensor imaging, DSI tractography resolves multiple myofiber populations per voxel, thus generating accurate 3D tractograms, which we present in the myocardium for the first time. METHODS AND RESULTS: DSI tractography was performed at 4.7 T in excised rat hearts 3 weeks after left coronary artery ligation (n=4) and in 4 age-matched controls. Fiber architecture in the control hearts varied smoothly from endocardium to epicardium, producing a symmetrical array of crossing helical structures in which orthogonal myofibers were separated by fibers with intermediate helix angles. Fiber architecture in the infarcted hearts was severely perturbed. The infarct boundary in all cases was highly irregular and punctuated repeatedly by residual myofibers extending from within the infarct to the border zones. In all infarcts, longitudinal myofibers extending toward the basal-anterior wall and transversely oriented myofibers extending toward the septum lay in direct contact with each other, forming nodes of orthogonal myofiber intersection or contact. CONCLUSIONS: DSI tractography resolves 3D myofiber architecture and reveals a complex network of orthogonal myofibers within infarcted myocardium. Meshlike networks of orthogonal myofibers in infarcted myocardium may resist mechanical remodeling but also probably increase the risk for lethal reentrant arrhythmias. DSI tractography thus provides a new and important readout of tissue injury after myocardial infarction. PMID- 19808595 TI - Applicability of appropriateness criteria for stress imaging: similarities and differences between stress echocardiography and single-photon emission computed tomography myocardial perfusion imaging criteria. AB - BACKGROUND: Appropriateness criteria for stress imaging have been published to promote the effective use of stress nuclear scintigraphy and stress echocardiography. We sought to evaluate the application of the stress echocardiography appropriateness criteria to patients undergoing stress echocardiography in an academic medical center. METHODS AND RESULTS: The stress echocardiography criteria were applied to 298 consecutive patients who underwent stress echocardiography. Patients were rated as appropriate, uncertain, inappropriate, or not classifiable. Results were compared with those of a previous analysis in the same patients using the single-photon computed tomography myocardial perfusion imaging (SPECT MPI) criteria. The level of agreement between 2 cardiac nurse abstractors for categorizing appropriateness by the stress echocardiography criteria was good (kappa=0.72). Overall, 54% of patients were classified as appropriate, 8% as uncertain, and 19% as inappropriate; 19% were not classifiable. By the SPECT MPI criteria, 64% of patients were classified as appropriate, 9% as uncertain, and 18% as inappropriate; 9% were not classifiable (P<0.001 compared with stress echocardiography criteria). By the stress echocardiography criteria, 6 clinical situations or indications accounted for more than 90% of the inappropriate tests; most of these involved asymptomatic patients. CONCLUSIONS: Applying stress echocardiography appropriateness criteria to a patient population is feasible, although 1 in 5 of our patients was not classifiable. Overall, the stress echocardiography criteria classified patients differently compared with the SPECT MPI criteria. Future refinements of the appropriateness criteria for stress imaging should address gaps in the criteria and disparities between the stress echocardiography and SPECT MPI criteria. PMID- 19808596 TI - Aortopulmonary collaterals after bidirectional cavopulmonary connection or Fontan completion: quantification with MRI. AB - BACKGROUND: Aortopulmonary collaterals (APCs) have been associated with increased morbidity after the Fontan operation. We aimed to quantify APC flow after bidirectional cavopulmonary connections and Fontan completions, using phase contrast MRI, and to identify risk factors for the development of APCs. METHODS AND RESULTS: APC blood flow was quantifiable in 24 of 36 retrospectively analyzed MRI studies. Sixteen studies were performed after the bidirectional cavopulmonary connections (group A) and 8 after the Fontan operation (group B). APC blood flow was calculated by subtracting the blood flow volume through the pulmonary arteries from that through the pulmonary veins. The ratio of pulmonary to systemic blood flow (Qp/Qs) was 0.93+/-0.26 in group A and 1.27+/-0.16 in group B. APC flow was 1.42 (0.58 to 3.83) L/min/m(2) and 0.82 (0.50 to 1.81) L/min/m(2) in groups A and B, respectively. The mean inaccuracies corresponded to 7.9+/ 14.5% and 7.1+/-13.6% of ascending aortic flow in groups A and B, respectively. Qp/Qs was negatively correlated with a younger age at the time of the bidirectional cavopulmonary connections operation (r=0.62, P=0.01) and positively correlated with the age at the time of the Fontan completion (r=0.81, P=0.01). Patients with a previous right-sided modified Blalock-Taussig shunt had more collateral flow to the right lung than those without. CONCLUSIONS: APC blood flow can be noninvasively measured in bidirectional cavopulmonary connections and Fontan patients, using MRI in the majority of patients and results in a significant left-to-right shunt. PMID- 19808597 TI - Comparison of gadofluorine-M and Gd-DTPA for noninvasive staging of atherosclerotic plaque stability using MRI. AB - BACKGROUND: Inflammation and neovascularization play critical roles in the stability of atherosclerotic plaques. Whole-body quantitative assessment of these plaque features may improve patient risk-stratification for life-threatening thromboembolic events and direct appropriate intervention. In this report, we determined the utility of the MR contrast agent gadofluorine-M (GdF) for staging plaque stability and compared this to the conventional agent Gd-DTPA. METHODS AND RESULTS: Five control and 7 atherosclerotic rabbits were sequentially imaged after administration of Gd-DTPA (0.2 mmol/kg) and GdF (0.1 mmol/kg) using a T(1) weighted pulse sequence on a 3-T MRI scanner. Diseased aortic wall could be distinguished from normal wall based on wall-to-muscle contrast-to-noise values after GdF administration. RAM-11 (macrophages) and CD-31 (endothelial cells) immunostaining of MR-matched histological sections revealed that GdF accumulation was related to the degree of inflammation at the surface of plaques and the extent of core neovascularization. Importantly, an MR measure of GdF accumulation at both 1 and 24 hours after injection but not Gd-DTPA at peak enhancement was shown to correlate with a quantitative histological morphology index related to these 2 plaque features. CONCLUSIONS: GdF-enhanced MRI of atherosclerotic plaques allows noninvasive quantitative information about plaque composition to be acquired at multiple time points after injection (within 1 and up to 24 hours after injection). This dramatically widens the imaging window for assessing plaque stability that is currently attainable with clinically approved MR agents, therefore opening the possibility of whole-body (including coronary) detection of unstable plaques in the future and potentially improved mitigation of cataclysmic cardiovascular events. PMID- 19808598 TI - In vivo quantification of carotid artery wall dimensions: 3.0-Tesla MRI versus B mode ultrasound imaging. AB - BACKGROUND: Our aim was to compare common carotid mean wall thickness (MWT) measurements by 3.0-T MRI with B-mode ultrasound common carotid intima-media thickness (CCIMT) measurements, a validated surrogate marker for cardiovascular disease. METHODS AND RESULTS: B-mode ultrasound and 3.0-T MRI scans of the left and right common carotid arteries were repeated 3 times in 15 healthy younger volunteers (age, 26+/-2.6 years), 15 healthy older volunteers (age, 57+/-3.2 years), and 15 subjects with cardiovascular disease and carotid atherosclerosis (age, 63+/-9.8 years). MWT was 0.711 (SD, 0.229) mm and mean CCIMT was 0.800 (SD, 0.206) mm. MWT and CCIMT were highly correlated (r=0.89, P<0.001). The intraclass correlation coefficients for interscan and interobserver and intraobserver agreements of MRI MWT measurements were larger than 0.95 with small confidence intervals, indicating excellent reproducibility. Power calculations indicate that 89 subjects are required to detect a 4% difference in MRI MWT compared with 469 subjects to detect similar differences with ultrasound IMT in follow-up studies. CONCLUSIONS: The study data for carotid MRI and ultrasound IMT showed strong agreement, indicating that both modalities measure the thickness of the intima and media. The advantage of MRI over ultrasound is that the measurement variability is smaller, enabling smaller sample sizes and potentially shorter study duration in cardiovascular prevention trials. PMID- 19808599 TI - The prognostic implications of cardiovascular magnetic resonance. PMID- 19808600 TI - Is computed tomography coronary angiography the most accurate and effective noninvasive imaging tool to evaluate patients with acute chest pain in the emergency department?: CT coronary angiography is the most accurate and effective noninvasive imaging tool for evaluating patients presenting with chest pain to the emergency department. PMID- 19808601 TI - Is computed tomography coronary angiography the most accurate and effective noninvasive imaging tool to evaluate patients with acute chest pain in the emergency department? CT coronary angiography is the most accurate and effective noninvasive imaging tool for evaluating patients presenting with chest pain to the emergency department: antagonist viewpoint. PMID- 19808602 TI - Cardiovascular images. Bland-White-Garland syndrome discovered in an elderly man. PMID- 19808603 TI - Cardiovascular images. A novel real-time fluorescent optical imaging system in mouse heart: a powerful tool for studying coronary circulation and cardiac function. PMID- 19808604 TI - Varix of the heart. PMID- 19808605 TI - Myocardial metastasis or benign brown fat? PMID- 19808606 TI - Right ventricular edema complicating acute inferior myocardial infarction as demonstrated by T2-weighted cardiovascular magnetic resonance. PMID- 19808607 TI - One more step for computed tomography coronary angiography before heart valve surgery. PMID- 19808608 TI - Left atrial volume and geometry in healthy aging: the Cardiovascular Health Study. AB - BACKGROUND: The left atrium is a validated marker of clinical and subclinical cardiovascular disease. Left atrial enlargement is often seen among older individuals; however, there are few population-based data regarding normal left atrial size among older persons, especially from those who are healthy, and from women. Furthermore, because the left atrium is a 3D structure, the commonly used parasternal long-axis diastolic diameter often underdiagnoses left atrial enlargement. METHODS AND RESULTS: We evaluated left atrial size in 230 healthy participants (mean age, 76+/-5 years) free of prevalent cardiac disease, rhythm abnormality, hypertension, and diabetes selected from the Cardiovascular Health Study, a prospective community-based study of risk factors for cardiovascular disease in 5888 elderly participants. In addition to the standard long-axis measurement, we obtained left atrial superoinferior and lateral diameters and used these dimensions to estimate left atrial volume. These measurements were used to generate reference ranges for determining left atrial enlargement in older men and women, based on the 95% percentiles of the left atrial dimensions in healthy participants, both unadjusted, and after adjustment for age, height, and weight. In healthy elderly subjects, indices of left atrial size do not correlate with age or height but with weight and other measures of body build. CONCLUSIONS: These data provide normative reference values for left atrial size in healthy older women and men. The results should be useful for refining diagnostic criteria for left atrial dilation in the older population and may be relevant for cardiovascular risk stratification. PMID- 19808609 TI - Are shades of gray prognostically useful in reporting myocardial perfusion single photon emission computed tomography? AB - BACKGROUND: We have advocated the use of a 5-category "normal," "probably normal," "equivocal," "probably abnormal," and "definitely abnormal" approach to final interpretation of myocardial perfusion single-photon emission computed tomography (SPECT). The prognostic value of expressing levels of certainty compared with a dichotomous normal/abnormal classification or categories for summed stress scores is unclear. METHODS AND RESULTS: Myocardial perfusion SPECT (MPS) was visually assessed using a standard semiquantitative approach, yielding summed scores that were used for preliminary interpretation using 5 levels of certainty. The interpreter was permitted to then shift the level of certainty in the final interpretation by 1 degree, based on nonperfusion MPS variables and available clinical information. To examine the prognostic value of expressing levels of clinical certainty, we evaluated 20 740 unique consecutive patients who underwent rest Tl-201/stress Tc-99m sestamibi MPS (34.3% vasodilator stress), of whom 845 (4.4%) were lost to follow-up and 1695 were excluded from prognostic analysis due to an early revascularization (<60 days after MPS). The remaining 18 200 patients (59.1% men; age, 65+/-13 years) were followed up for cardiac death for a mean of 2.7+/-1.7 years. During the follow-up, a total of 591 cardiac death events occurred. By univariable analysis, there were substantial differences in the distribution of follow-up cardiac death by the category of clinical MPS certainty. The clinical certainty was found to be an independent multivariable predictor of cardiac death in the study population and better identified patients at increased risk of cardiac death than the approaches based solely on the standard categories of summed perfusion scores or based solely on categories of segmental perfusion scores. CONCLUSIONS: The use of multicategory reporting of MPS results incorporating nonperfusion MPS results and clinical information enhances risk stratification compared with both a dichotomous normal/abnormal approach or approaches based solely on segmental categories of perfusion scores. Whether this enhanced risk stratification based on the clinical certainty of the MPS interpretation leads to a more effective therapeutic regimen, tailored to the individual patient's need, requires further prospective evaluation. PMID- 19808610 TI - Integration of infarct size, tissue perfusion, and metabolism by hybrid cardiac positron emission tomography/computed tomography: evaluation in a porcine model of myocardial infarction. AB - BACKGROUND: Hybrid positron emission tomography/computed tomography (PET-CT) allows for combination of PET perfusion/metabolism imaging with infarct detection by CT delayed contrast enhancement. We used this technique to obtain biomorphological insights into the interrelation between tissue damage, inflammation, and microvascular obstruction early after myocardial infarction. METHODS AND RESULTS: A porcine model of left anterior descending coronary artery occlusion/reperfusion was studied. Seven animals underwent PET-CT within 3 days of infarction, and a control group of 3 animals was scanned at >4 weeks. Perfusion and glucose uptake were assessed by [(13)N]-ammonia/[(18)F] deoxyglucose (FDG), and 64-slice CT delayed contrast enhancement was measured. In the acute infarct model, CT revealed a no-reflow phenomenon suggesting microvascular obstruction in 80% of all infarct segments. PET showed increased FDG uptake in 68% of the CT-defined infarct segments. Ex vivo staining and histology showed active inflammation in the acute infarct area as an explanation for increased glucose uptake. In chronic infarction, CT showed no microvascular obstruction and agreed well with matched perfusion/metabolism defects on PET. CONCLUSIONS: Perfusion/metabolism PET and delayed enhancement CT can be combined within a single hybrid PET-CT session. Increased regional FDG uptake in the acute infarct area is frequently observed. In contrast to the chronic infarct setting, this indicates tissue inflammation that is commonly associated with microvascular obstruction as identified by no reflow on CT. The consequences of these pathophysiological findings for subsequent ventricular remodeling should be explored in further studies. PMID- 19808611 TI - Multislice computed tomography in the exclusion of coronary artery disease in patients with presurgical valve disease. AB - BACKGROUND: Multislice computed tomography (MSCT) has shown high negative predictive value in ruling out obstructive coronary artery disease. Preliminary studies in patients with valvular heart disease (VHD) have demonstrated the potential of MSCT angiography (CTA) in such patients, precluding need for invasive angiography (XA). However, larger prospectively designed studies, including patients with atrial fibrillation and incorporating dose reduction algorithms, are needed. METHODS AND RESULTS: To evaluate the clinical utility of 64-slice CT in the preoperative assessment in patients with VHD, we prospectively studied 452 consecutive patients undergoing routine cardiac catheterization for eligibility. Two hundred thirty-seven patients underwent both MSCT and XA. Segment-based, vessel-based, and patient-based agreement between CTA and XA was estimated assuming that "nonevaluable" segments were positive for significant coronary stenosis. In a patient-based analysis, sensitivity, specificity, positive predictive value, and negative predictive values of CTA were 95%, 89%, 66%, and 99%, respectively; in vessel-based analysis, 90%, 92%, 48%, and 99%, respectively; and in segment-based analysis, 89%, 97%, 38%, and 100%, respectively. No significant differences were found between patients with or without atrial fibrillation. A CAC value of 390 was the best cutoff for the identification of patients with positive or inconclusive CTA (which would not be exempted from XA in the clinical setting). CONCLUSIONS: In the preoperative assessment of patients with predominant VHD, the diagnostic accuracy of 64-slice CTA for ruling out the presence of significant coronary artery disease is very good even when including patients with irregular heart rhythm. Using this approach, CAC quantification before CTA can be successfully used to identify patients who should be referred directly to XA, sparing unnecessary exposure to radiation. PMID- 19808612 TI - Correlates of carotid plaque presence and composition as measured by MRI: the Atherosclerosis Risk in Communities Study. AB - BACKGROUND: The composition of atherosclerotic plaque affects the likelihood of an atherothrombotic event, but prospective studies relating risk factors to carotid wall and plaque characteristics measured by MRI are lacking. We hypothesized that traditional risk factors are predictors of carotid wall and plaque characteristics measured 2 decades later. METHODS AND RESULTS: A high resolution contrast-enhanced MRI examination of the carotid artery was performed in 1769 participants. Measures of carotid wall volume and maximum thickness; lipid core presence, volume and maximum area; and fibrous cap thickness were performed centrally. The sample was, on average, 70 years of age, 57% female, 81% white, and 19% black. Greater age, total and low-density lipoprotein cholesterol, male sex, white race, diabetes, hypertension, and smoking as measured at baseline were all significant predictors of increased wall volume and maximum wall thickness 18 years later. An analysis of lipid core was restricted to the 1180 participants with maximum wall thickness >/=1.5 mm. Lipid core was observed in 569 individuals (weighted percentage, 42%). Baseline age and total and low density lipoprotein cholesterol were predictors of presence of lipid core 18 years later; however, these relationships were attenuated after adjustment for wall thickness. Concurrently measured low-density lipoprotein cholesterol was associated with greater lipid core volume, independent of wall thickness. Concurrently measured glucose and body mass index were inversely associated fibrous cap thickness. CONCLUSIONS: Traditional atherosclerosis risk factors are related to increased wall volume and wall thickness 2 decades later, but they do not discriminate characteristics of plaque composition (core and cap) independent of wall size. PMID- 19808613 TI - Alteration in left ventricular strains and torsional mechanics after ultralong duration exercise in athletes. AB - BACKGROUND: Numerous studies have reported evidence of cardiac injury associated with transient left ventricular (LV) systolic and diastolic dysfunction after prolonged and strenuous exercise. We used 2D ultrasound speckle tracking imaging to evaluate the effect of an ultralong-duration exercise on LV regional strains and torsion. We speculated that systolic dysfunction after exercise is associated with depressed LV strains and torsion, and diastolic dysfunction results from decreased and delayed untwisting, a key factor of LV suction and early filling. METHODS AND RESULTS: Twenty-three triathletes underwent conventional and speckle tracking imaging echocardiography at rest before and immediately after an ultralong distance triathlon. Measurements included LV longitudinal, circumferential and radial strains, LV rotations, and LV torsion. After the race, LV systolic dysfunction was characterized by a decrease in LV longitudinal, radial, and circumferential strains, especially for apical radial strains (44.6+/ 15.1% versus 31.1+/-13.8%, P<0.001). Peak torsion was slightly decreased (8.3+/ 5.1 degrees versus 6.4+/-3.9 degrees , respectively, P=0.09) and significantly delayed (91+/-18% versus 128+/-31% of systolic duration, P<0.001) beside end ejection. Peak untwisting was also depressed and delayed beside isovolumic relaxation. CONCLUSIONS: This study documented major alterations in cardiac strains and torsion after an ultralong distance triathlon. LV systolic strains were depressed but not delayed, whereas twisting was decreased and delayed. This altered pattern hampered the rapid untwisting during isovolumic relaxation phase, reducing LV diastolic suction and early filling. PMID- 19808614 TI - Evaluation of alphavbeta3 integrin-targeted positron emission tomography tracer 18F-galacto-RGD for imaging of vascular inflammation in atherosclerotic mice. AB - BACKGROUND: (18)F-Galacto-RGD is a positron emission tomography (PET) tracer binding to alpha(v)beta(3) integrin that is expressed by macrophages and endothelial cells in atherosclerotic lesions. Therefore, we evaluated (18)F galacto-RGD for imaging vascular inflammation by studying its uptake into atherosclerotic lesions of hypercholesterolemic mice in comparison to deoxyglucose. METHODS AND RESULTS: Hypercholesterolemic LDLR(-/-)ApoB(100/100) mice on a Western diet and normally fed adult C57BL/6 control mice were injected with (18)F-galacto-RGD and (3)H-deoxyglucose followed by imaging with a small animal PET/CT scanner. The aorta was dissected 2 hours after tracer injection for biodistribution studies, autoradiography, and histology. Biodistribution of (18)F galacto-RGD was higher in the atherosclerotic than in the normal aorta. Autoradiography demonstrated focal (18)F-galacto-RGD uptake in the atherosclerotic plaques when compared with the adjacent normal vessel wall or adventitia. Plaque-to-normal vessel wall ratios were comparable to those of deoxyglucose. Although angiogenesis was not detected, (18)F-galacto-RGD uptake was associated with macrophage density and deoxyglucose accumulation in the plaques. Binding to atherosclerotic lesions was efficiently blocked in competition experiments. In vivo imaging visualized (18)F-galacto-RGD uptake colocalizing with calcified lesions of the aortic arch as seen in CT angiography. CONCLUSIONS: (18)F-Galacto-RGD demonstrates specific uptake in atherosclerotic lesions of mouse aorta. In this model, its uptake was associated with macrophage density. (18)F-Galacto-RGD is a potential tracer for noninvasive imaging of inflammation in atherosclerotic lesions. PMID- 19808615 TI - Outcomes research in cardiovascular imaging: report of a workshop sponsored by the National Heart, Lung, and Blood Institute. AB - In July of 2008, the National Heart, Lung, and Blood Institute convened experts in noninvasive cardiovascular imaging, outcomes research, statistics, and clinical trials to develop recommendations for future randomized controlled trials of the use of imaging in: 1) screening the asymptomatic patient for coronary artery disease; 2) assessment of patients with stable angina; 3) identification of acute coronary syndromes in the emergency room; and 4) assessment of heart failure patients with chronic coronary artery disease with reduced left ventricular ejection fraction. This study highlights several possible trial designs for each clinical situation. PMID- 19808616 TI - Extensive myocardial fibrosis in a patient with hypertrophic cardiomyopathy and ventricular tachycardia without traditional high-risk features. PMID- 19808617 TI - Primary stenting of an anomalous left main coronary artery with an interarterial course during cardiac arrest: imaging with CT angiography. PMID- 19808618 TI - Cough: a potentially life-threatening condition after interventional closure of atrial septal defect. PMID- 19808619 TI - Echocardiographic and macroscopic images: aortic cusp laceration. PMID- 19808620 TI - Absent right superior vena cava: multimodality imaging of upper body venous drainage via left-sided superior vena cava and azygos venous system. PMID- 19808621 TI - MRI of intimal sarcoma of the pulmonary arteries. PMID- 19808622 TI - Imaging left ventricular muscarinic receptor heterogeneity: a tool to evaluate individuals at risk for sudden death? PMID- 19808623 TI - Prediction of all-cause mortality from global longitudinal speckle strain: comparison with ejection fraction and wall motion scoring. AB - BACKGROUND: Although global left ventricular systolic function is an important determinant of mortality, standard measures such as ejection fraction (EF) and wall motion score index (WMSI) have important technical limitations. The aim of this study was to compare global longitudinal speckle strain (GLS), an automated technique for measurement of long-axis function, with EF and WMSI for the prediction of mortality. METHODS AND RESULTS: Of 546 consecutive individuals undergoing echocardiography for assessment of resting left ventricular function, 91 died over a period of 5.2+/-1.5 years. In addition to Simpson biplane EF, WMSI was determined by 2 experienced readers and GLS was calculated from 3 standard apical views using 2D speckle tracking. The incremental value of EF, WMSI, and GLS to significant clinical variables was assessed in nested Cox models. Clinical factors associated with outcome (model chi(2)=20.2) were age (hazard ratio [HR], 1.46; P<0.01), diabetes (HR, 1.88; P=0.01), and hypertension (HR, 1.59; P<0.05). Although addition of EF (HR, 1.23; P=0.03) or WMSI (HR, 1.28; P<0.01) added to the predictive power of clinical variables, the addition of GLS (HR, 1.45; P<0.001) caused the greatest increment in model power (chi(2)=34.9, P<0.001). GLS also provided incremental value in subgroups with EF >35% and those with and without wall motion abnormalities. A GLS >or=-12% was found to be equivalent to an EF 70% diameter reduction arteries and without signs of damage; and damaged, with damage. The muscarinic receptor density in remote (67+/ 30 pmol/mL tissue; n=86) and potentially damaged (71+/-30 pmol/mL tissue; n=42) regions of patients was higher than in normal regions of volunteers (32+/-17 pmol/mL tissue; n=156; P<0.001). The muscarinic receptor density in damaged regions (42+/-21 pmol/mL tissue; n=58) was reduced compared with remote and potentially damaged regions (P<0.001) but was not significantly different from normal regions in volunteers (P=0.093). CONCLUSIONS: Vagal control in patients with chronic myocardial infarction involves muscarinic receptor upregulation in remote nondamaged left ventricular regions. Our results suggest that the receptor density remains within normal values in myocardial regions containing damaged tissue. PMID- 19808625 TI - In vivo metabolic phenotyping of myocardial substrate metabolism in rodents: differential efficacy of metformin and rosiglitazone monotherapy. AB - BACKGROUND: Cardiovascular disease is the leading cause of death among diabetic patients, with alteration in myocardial substrate metabolism being a likely contributor. We aimed to assess noninvasively the efficacy of metformin and rosiglitazone monotherapy in normalizing myocardial substrate metabolism in an animal model of type 2 diabetes mellitus. METHODS AND RESULTS: The study used 18 male ZDF rats (fa/fa) with 6 rats in each group: an untreated group; a group treated with metformin (16.6 mg/kg/d), and a group treated with rosiglitazone (4 mg/kg). Each rat was scanned at age 14 weeks (baseline) and subsequently at 19 weeks with small-animal positron emission tomography to estimate myocardial glucose utilization (MGU) and myocardial utilization (MFAU), oxidation (MFAO), and esterification (MFAE). Treatment lasted for 5 weeks after baseline imaging. At week 19, rats were euthanized and hearts were extracted for expression analysis of select genes encoding for GLUT transporters and fatty acid transport and oxidation genes. In addition, echocardiography measurements were obtained at weeks 13 and 18 to characterize cardiac function. Metformin had no significant effect on either MGU or MFAU and MFAO. In contrast, rosiglitazone tended to enhance MGU and significantly reduced MFAU and MFAO. Rosiglitazone-induced increase in glucose uptake correlated significantly with increased expression of GLUT4, whereas diminished MFAO correlated significantly with decreased expression of FATP-1 and MCAD. Finally, changes in fractional shortening as a measure of cardiac function were unchanged throughout the study. CONCLUSIONS: Treatment with rosiglitazone enhanced glucose utilization and diminished MFAO, thus reversing the metabolic phenotype of the diabetic heart. PMID- 19808626 TI - Cardiac mechanics in mild hypertensive heart disease: a speckle-strain imaging study. AB - BACKGROUND: We hypothesized that abnormalities in regional systolic strain (epsilon) might be present among hypertensive subjects with normal ejection fraction, and, if present, could be used to identify patients at high risk for heart failure. The aim of the current case-control study was to use speckle tracking imaging to identify subclinical global and regional systolic function abnormalities in hypertensive subjects with normal ejection fraction. METHODS AND RESULTS: Standard 2D Doppler echocardiography, tissue Doppler imaging, and 2D speckle strain imaging were performed in 52 hypertensive subjects with normal ejection fraction and 52 control subjects of similar age. Peak systolic (S'), and diastolic (E') annular velocities were obtained by tissue Doppler imaging, whereas longitudinal myocardial systolic velocity (V(l)) and circumferential, longitudinal, and radial strains (epsilon(c), epsilon(l), epsilon(r)) were obtained by speckle tracking. Midwall shortening and peak basal longitudinal strain (epsilon(l)) were used as indices of regional function. Hypertensive subjects had lower velocities--tissue Doppler imaging E' and S', and V(l)--and evidence of reduced regional function. Surprisingly, however, global epsilon values did not differentiate hypertensive subjects from control subjects. Among hypertensive patients, significant inverse associations were found between left ventricular mass and global longitudinal and circumferential epsilon (both P<0.05). CONCLUSIONS: Hypertensive heart disease with normal ejection fraction is associated with reduced myocardial velocities and reduced regional function but normal global epsilon. Our data suggest that velocity abnormalities occur early in hypertension and may be an appropriate target for preventive strategies because they occur before abnormalities in global epsilon. PMID- 19808627 TI - Increased neovascularization in advanced lipid-rich atherosclerotic lesions detected by gadofluorine-M-enhanced MRI: implications for plaque vulnerability. AB - BACKGROUND: Inflammation and neovascularization may play a significant role in atherosclerotic plaque progression and rupture. We evaluated gadofluorine-M enhanced MRI for detection of plaque inflammation and neovascularization in an animal model of atherosclerosis. METHODS AND RESULTS: Sixteen rabbits with aortic plaque and 6 normal control rabbits underwent gadofluorine-M-enhanced MRI. Eight rabbits had advanced atherosclerotic lesions, whereas the remaining 8 had early lesions. Magnetic resonance atherosclerotic plaque enhancement was meticulously compared with plaque inflammation and neovessel density as assessed by histopathology. Advanced plaques and early atheroma were enhanced after gadofluorine-M injection. Control animals displayed no enhancement. After accounting for the within-animal correlation of observations, mean contrast-to noise ratio was significantly higher in advanced plaques than compared with early atheroma (4.29+/-0.21 versus 3.00+/-0.32; P=0.004). Macrophage density was higher in advanced plaques in comparison to early atheroma (geometric mean=0.50 [95% CI, 0.19 to 1.03] versus 0.25 [0.07 to 0.42]; P=0.05). Furthermore, higher neovessel density was observed in advanced plaques (1.83 [95% CI, 1.51 to 2.21] versus 1.29 [0.99 to 1.69]; P=0.05). The plaque accumulation of gadofluorine-M correlated with increased neovessel density as shown by linear regression analysis (r=0.67; P<0.001). Confocal and fluorescence microscopy revealed colocalization of gadofluorine-M with plaque areas containing a high density of neovessels. CONCLUSIONS: Gadofluorine-M-enhanced MRI is effective for in vivo detection of atherosclerotic plaque inflammation and neovascularization in an animal model of atherosclerosis. These findings suggest that gadofluorine-M enhancement reflects the presence of high-risk plaque features believed to be associated with plaque rupture. Gadofluorine-M plaque enhancement may therefore provide functional assessment of atherosclerotic plaque in vivo. PMID- 19808628 TI - Feasibility of cardiovascular magnetic resonance to assess the orifice area of aortic bioprostheses. AB - BACKGROUND: Prosthetic orifice area, usually calculated by transthoracic echocardiography (TTE) or transesophageal echocardiography (TEE), provides important information regarding the hemodynamic performance of aortic bioprostheses. However, both TTE and TEE have limitations; therefore accurate and reproducible determination of the orifice area often remains a challenge. The present study aimed to investigate the feasibility of cardiovascular magnetic resonance (CMR) to assess the orifice areas of aortic bioprostheses. METHODS AND RESULTS: CMR planimetry of the orifice area was performed in 65 patients (43/22 stented/stentless prostheses; mean time since implantation, 3.1+/-2.8 years; mean orifice area [TTE], 1.70+/-0.43 cm(2); 62 normally functioning prostheses, 2 severe stenoses, and 1 severe regurgitation) in an imaging plane perpendicular to the transprosthetic flow using steady-state free-precession cine imaging under breath-hold conditions on a 1.5-T MR system. CMR results were compared with TTE (continuity equation, n=65) and TEE (planimetry, n=31). CMR planimetry was readily feasible in 80.0%; feasible with limitation in 15.4% because of stent, flow, and sternal wire artifacts; and impossible in 4.6% because of flow artifacts. Correlations of the orifice areas by CMR with TTE (r=0.82) and CMR with TEE (r=0.92) were significant. The average difference between the methods was -0.02+/-0.24 cm(2) (TTE) and 0.05+/-0.15 cm(2) (TEE). Agreement was present for stented and stentless devices and independent of orifice size. Intraobserver and interobserver variabilities of CMR planimetry were 6.7+/-5.4% and 11.5+/ 7.8%. CONCLUSIONS: The assessment of aortic bioprostheses with normal orifice areas by CMR is technically feasible and provides orifice areas with a close correlation to echocardiography and low observer dependency. PMID- 19808629 TI - Noninvasive quantification of systemic-to-pulmonary collateral flow: a major source of inefficiency in patients with superior cavopulmonary connections. AB - BACKGROUND: Systemic-to-pulmonary collateral flow (SPCF) is common in single ventricle patients with superior cavopulmonary connections (SCPC). Because no validated method to quantify that SPCF exists, neither its hemodynamic burden nor its clinical impact can be systematically evaluated. We hypothesize that (1) the difference in total ascending aortic (Ao) and caval flow (superior vena cava [SVC]+inferior vena cava [IVC]) and (2) the difference between pulmonary vein and pulmonary artery flow (PV-PA) provide 2 independent estimators of SPCF. METHODS AND RESULTS: We measured Ao, SVC, IVC, right (RPA) and left (LPA) PA, and left (LPV) and right (RPV) PV flows in 17 patients with SCPC during routine cardiac MRI studies using through-plane phase-contrast velocity mapping. Two independent measures of SPCF were obtained: model 1, Ao-(SVC+IVC); and model 2, (LPV LPA)+(RPV-RPA). Values were normalized to body surface area, Ao, and PV, and comparisons were made using linear regression and Bland-Altman analysis. SPCF ranged from 0.2 to 1.4 L/min for model 1 and 0.2 to 1.6 L/min for model 2, for an average indexed SPCF of 0.5 to 2.8 L/min/m(2): 11% to 53% (mean, 37%) of Ao and 19% to 77% (mean, 54%) of PV. The mean difference between model 1 and model 2 was 0.01 L/min (P=0.40; 2-SD range, -0.45 to 0.47 L/min). CONCLUSIONS: We present a noninvasive method for SPCF quantification in patients with SCPC. It should provide an important clinical tool in treating these patients. Furthermore, we show that SPCF is a significant hemodynamic burden in many patients with bidirectional Glenn shunt physiology. Future investigations will allow objective study of the impact of collateral flow on outcome. PMID- 19808630 TI - Noninvasive assessment of myocardial perfusion. PMID- 19808631 TI - Multimodality imaging assessment of anatomic and functional pulmonary vein stenosis. PMID- 19808632 TI - The aftermath of hip fracture: discharge placement, functional status change, and mortality. AB - The authors prospectively explored the consequences of hip fracture with regard to discharge placement, functional status, and mortality using the Survey on Assets and Health Dynamics Among the Oldest Old (AHEAD). Data from baseline (1993) AHEAD interviews and biennial follow-up interviews were linked to Medicare claims data from 1993-2005. There were 495 postbaseline hip fractures among 5,511 respondents aged >or=69 years. Mean age at hip fracture was 85 years; 73% of fracture patients were white women, 45% had pertrochanteric fractures, and 55% underwent surgical pinning. Most patients (58%) were discharged to a nursing facility, with 14% being discharged to their homes. In-hospital, 6-month, and 1 year mortality were 2.7%, 19%, and 26%, respectively. Declines in functional status-scale scores ranged from 29% on the fine motor skills scale to 56% on the mobility index. Mean scale score declines were 1.9 for activities of daily living, 1.7 for instrumental activities of daily living, and 2.2 for depressive symptoms; scores on mobility, large muscle, gross motor, and cognitive status scales worsened by 2.3, 1.6, 2.2, and 2.5 points, respectively. Hip fracture characteristics, socioeconomic status, and year of fracture were significantly associated with discharge placement. Sex, age, dementia, and frailty were significantly associated with mortality. This is one of the few studies to prospectively capture these declines in functional status after hip fracture. PMID- 19808633 TI - Editorial: Breathing new life into pneumonia epidemiology. PMID- 19808634 TI - Maternal licorice consumption and detrimental cognitive and psychiatric outcomes in children. AB - Overexposure to glucocorticoids may link prenatal adversity with detrimental outcomes in later life. Glycyrrhiza, a natural constituent of licorice, inhibits placental 11-beta-hydroxysteroid dehydrogenase type 2, the feto-placental "barrier" to higher maternal levels of cortisol. The authors studied whether prenatal exposure to glycyrrhiza in licorice exerts detrimental effects on cognitive performance (subtests of the Wechsler Intelligence Scale for Children III as well as the Children's Developmental Neuropsychological Assessment and the Beery Developmental Test of Visual-Motor Integration) and psychiatric symptoms (Child Behavior Checklist) in 321 Finnish children 8.1 years of age born in 1998 as healthy singletons at 35-42 weeks of gestation. In comparison to the group with zero-low glycyrrhiza exposure (0-249 mg/week), those with high exposure (>or=500 mg/week) had significant decrements in verbal and visuospatial abilities and in narrative memory (range of mean differences in standard deviation units, 0.31 to -0.41; P < 0.05) and significant increases in externalizing symptoms and in attention, rule-breaking, and aggression problems (range of odds ratios, 2.15 to 3.43; P < 0.05). The effects on cognitive performance appeared dose related. Data are compatible with adverse fetal "programming" by overexposure to glucocorticoids and caution against excessive intake of licorice-containing foodstuffs during pregnancy. PMID- 19808635 TI - Prospective study of urban form and physical activity in the Black Women's Health Study. AB - The authors used data from the Black Women's Health Study to assess the association between neighborhood urban form and physical activity. Women reported hours/week of utilitarian and exercise walking and of vigorous activity in 1995 and on biennial follow-up questionnaires through 2001. Housing density, road networks, availability of public transit, sidewalks, and parks were characterized for the residential neighborhoods of 20,354 Black Women's Health Study participants living in New York, New York; Chicago, Illinois; and Los Angeles, California. The authors quantified the associations between features of the environment and physical activity using odds ratios for >or=5 relative to <5 hours/week of physical activity. For all women, housing density had the strongest association with utilitarian walking (odds ratio for the most- compared with the least-dense quintile = 2.72, 95% confidence interval: 2.22, 3.31), followed by availability of public transit. Women who moved during follow-up to neighborhoods of lower density were 36% more likely to decrease their levels of utilitarian walking, and those who moved to neighborhoods of higher density were 23% more likely to increase their levels of utilitarian walking, relative to women who moved to neighborhoods of similar density. These data suggest that increases in housing density may lead to increases in utilitarian walking among African American women. PMID- 19808636 TI - Discovery properties of genome-wide association signals from cumulatively combined data sets. AB - Genetic effects for common variants affecting complex disease risk are subtle. Single genome-wide association (GWA) studies are typically underpowered to detect these effects, and combination of several GWA data sets is needed to enhance discovery. The authors investigated the properties of the discovery process in simulated cumulative meta-analyses of GWA study-derived signals allowing for potential genetic model misspecification and between-study heterogeneity. Variants with null effects on average (but also between-data set heterogeneity) could yield false-positive associations with seemingly homogeneous effects. Random effects had higher than appropriate false-positive rates when there were few data sets. The log-additive model had the lowest false-positive rate. Under heterogeneity, random-effects meta-analyses of 2-10 data sets averaging 1,000 cases/1,000 controls each did not increase power, or the meta-analysis was even less powerful than a single study (power desert). Upward bias in effect estimates and underestimation of between-study heterogeneity were common. Fixed-effects calculations avoided power deserts and maximized discovery of association signals at the expense of much higher false-positive rates. Therefore, random- and fixed effects models are preferable for different purposes (fixed effects for initial screenings, random effects for generalizability applications). These results may have broader implications for the design and interpretation of large-scale multiteam collaborative studies discovering common gene variants. PMID- 19808637 TI - Meat and meat-related compounds and risk of prostate cancer in a large prospective cohort study in the United States. AB - The authors examined associations between meat consumption (type, cooking method, and related mutagens), heme iron, nitrite/nitrate, and prostate cancer in a cohort of 175,343 US men aged 50-71 years. During 9 years of follow-up (1995 2003), they ascertained 10,313 prostate cancer cases (1,102 advanced) and 419 fatal cases. Hazard ratios comparing the fifth intake quintile with the first revealed elevated risks associated with red and processed meat for total (red meat: hazard ratio (HR) = 1.12, 95% confidence interval (CI): 1.04, 1.21; processed meat: HR = 1.07, 95% CI: 1.00, 1.14) and advanced (red meat: HR = 1.31, 95% CI: 1.05, 1.65; processed meat: HR = 1.32, 95% CI: 1.08, 1.61) prostate cancer. Heme iron, barbecued/grilled meat, and benzo[a]pyrene were all positively associated with total (HR = 1.09 (95% CI: 1.02, 1.17), HR = 1.11 (95% CI: 1.03, 1.19), and HR = 1.09 (95% CI: 1.00, 1.18), respectively) and advanced (HR = 1.28 (95% CI: 1.03, 1.58), HR = 1.36 (95% CI: 1.10, 1.69), and HR = 1.28 (95% CI: 1.00, 1.65), respectively) disease. Nitrite (HR = 1.24, 95% CI: 1.02, 1.51) and nitrate (HR = 1.31, 95% CI: 1.07, 1.61) intakes were associated with advanced prostate cancer. There were no clear associations for fatal prostate cancer. Red and processed meat may be positively associated with prostate cancer via mechanisms involving heme iron, nitrite/nitrate, grilling/barbecuing, and benzo[a]pyrene. PMID- 19808641 TI - Polypodium leucotomos extract decreases UV-induced Cox-2 expression and inflammation, enhances DNA repair, and decreases mutagenesis in hairless mice. AB - UV-irradiated skin and UV-induced tumors overexpress the inducible isoform of cyclooxygenase-2 (Cox-2), and Cox-2 inhibition reduces photocarcinogenesis. To evaluate photoprotective effects of Polypodium leucotomos extract (PL), hairless Xpc(+/-) mice were fed for 10 days with PL (300 mg/kg) or vehicle then UV irradiated, once. By 24 hours, UV-induced Cox-2 levels were increased in vehicle fed and PL-fed mice, whereas by 48 and 72 hours, Cox-2 levels were four- to fivefold lower in PL-fed mice (P < 0.05). p53 expression/activity was increased in PL-fed versus vehicle-fed then UV-irradiated mice. UV-induced inflammation was decreased in PL-fed mice, as shown by approximately 60% decrease (P < 0.001) in neutrophil infiltration at 24 hours, and macrophages by approximately 50% (<0.02) at 24 and 48 hours. By 72 hours, 54 +/- 5% cyclobutane pyrimidine dimers remained in vehicle-fed versus 31 +/- 5% in PL-fed skin (P < 0.003). The number of 8 hydroxy-2'-deoxyguanosine-positive cells were decreased before UV irradiation by approximately 36% (P < 0.01), suggesting that PL reduces constitutive oxidative DNA damage. By 6 and 24 hours, the number of 8-hydroxy-2'-deoxyguanosine-positive cells were approximately 59% (P < 0.01) and approximately 79% (P < 0.03) lower in PL-fed versus vehicle-fed mice. Finally, UV-induced mutations in PL-fed-mice were decreased by approximately 25% when assessed 2 weeks after the single UV exposure. These data demonstrate that PL extract supplementation affords the following photoprotective effects: p53 activation and reduction of acute inflammation via Cox-2 enzyme inhibition, increased cyclobutane pyrimidine dimer removal, and reduction of oxidative DNA damage. PMID- 19808642 TI - Contribution of macrophages to angiogenesis induced by vascular endothelial growth factor receptor-3-specific ligands. AB - Vascular endothelial growth factor receptor (VEGFR)-2 is a major stimulator of hemangiogenesis (HA), whereas VEGFR-3 stimulates lymphangiogenesis (LA). Contrary to this understanding, we demonstrate that implantation of pellets containing VEGFR-3-specific ligands (VEGF-C156S and recombinant murine VEGF-D) into the corneal stroma induce not only LA but also robust HA characterized by blood vessels that are positive for VEGFR-3 expression. The implantation of pellets containing VEGFR-3-specific ligands also leads to the recruitment of VEGF-A secreting macrophages. Depletion of these infiltrating macrophages using clodronate-liposome administration shows a significant reduction in HA as well as LA. Blockade of either VEGFR-2 or VEGFR-3 signaling reduces both HA and LA; however, the percent reduction of HA is greater in the VEGFR-2 blockade group. In addition, in the VEGFR-3 blockade group, the percent reduction of HA is significantly greater with VEGFR-3-specific ligands than that by VEGF-A or VEGF C. Collectively, our data suggest that VEGFR-3-specific signaling can induce new blood vessels, to which macrophages contribute a major role, and signify its potential as an additional therapeutic target to the existing VEGF-A/VEGFR-2 signaling-based antiangiogenesis strategies. PMID- 19808643 TI - Subcellular localization of the HER4 intracellular domain, 4ICD, identifies distinct prognostic outcomes for breast cancer patients. AB - Conflicting reports of the prognostic value of HER4 in breast cancer may be explained by distinct activities of the HER4 intracellular domain, 4ICD. Here, immunohistochemical 4ICD staining of archival invasive breast cancers (n = 923) was scored separately for nuclear and cytosolic expression, and these data were tested for associations with clinicopathological markers, disease-free survival, and disease-specific survival. By univariate analysis, cytosolic 4ICD expression was independently associated with estrogen receptor and progesterone receptor expression and tumor cell apoptosis. Nuclear 4ICD inversely correlated with tumor grade and tumor mitosis. In multivariate analyses cytosolic, but not nuclear 4ICD, significantly correlated with disease-free survival (P = 0.035) and disease specific survival (P < 0.004) in lymph node-negative patients. Our results demonstrate for the first time that cytosolic 4ICD has significant positive prognostic value in node-negative breast cancer patients. At present, tumor grade and size are the primary clinicopathological parameters commonly used to guide decision making in these patients. Our results suggest that cytosolic 4ICD has important pathological functions and may be used to identify node-negative breast cancer patients at low risk of relapse and an improved survival, thereby avoiding systemic overtreatment of these patients. Our results also suggest that pan receptor tyrosine kinase inhibitors, currently in clinical trials, or HER4 antagonists, which disengage 4ICD signaling, may have untoward activity in patients whose tumors express cytosolic 4ICD. PMID- 19808644 TI - {beta}3-integrin expression on tumor cells inhibits tumor progression, reduces metastasis, and is associated with a favorable prognosis in patients with ovarian cancer. AB - The role of the vitronectin receptor (alpha(v)beta(3)-integrin) as a tumor promoter seems well established, and, consequently, therapies that block this integrin are currently in clinical testing. We undertook the current study to determine whether alpha(v)beta(3)-integrin is an appropriate target in ovarian cancer treatment. Expression of beta(3)-integrin in SKOV3ip1 ovarian cancer cells led to the overexpression of alpha(v)beta(3)-integrin on the cell surface and increased adhesion. However, alpha(v)beta(3)-integrin-overexpressing cells showed impaired invasion, protease expression, and colony formation. These results were recapitulated in xenograft studies: alpha(v)beta(3)-integrin-expressing cells showed increased adhesion to mouse peritoneum, but the overall number of metastatic nodules (105 versus 68 tumors) and tumor weight were significantly lower than those in the parental SKOV3ip1 cells. The alpha(v)beta(3)-integrin overexpressing cells had a decreased proliferation rate mediated by inhibition of cyclin B1 and induction of phospho-Cdc2 and p53 expression, consistent with a G(2)M cell cycle arrest. Confirming the above results, inhibition of beta(3) integrin in cultured or primary OvCa cells decreased adhesion but increased invasion and proliferation. Patients with tumors expressing high beta(3)-integrin had significantly better disease-free and overall survival (52 months versus 27 months, P < 0.05). This study shows that alpha(v)beta(3)-integrin expression on tumor cells actually slows tumor progression and acts as a tumor suppressor. Therefore, the vitronectin receptor might not be an appropriate therapeutic target in ovarian cancer. PMID- 19808645 TI - Transcription factors Kruppel-like factor 6 and peroxisome proliferator-activated receptor-{gamma} mediate high glucose-induced thioredoxin-interacting protein. AB - We demonstrated recently that thioredoxin-interacting protein (Txnip) and the transcription factor Kruppel-like factor 6 (KLF6) were up-regulated in both in vivo and in vitro models of diabetic nephropathy, thus promoting renal injury. Conversely, peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonists have been shown to be renoprotective. Hence, this study was undertaken to determine whether Txnip expression is regulated by the transcription factors KLF6 and PPAR-gamma. By using siRNAs and overexpressing constructs, the role of KLF6 and PPAR-gamma in Txnip transcriptional regulation was determined in human kidney proximal tubule cells and in streptozocin-induced diabetes mellitus in Sprague-Dawley rats, in vitro and in vivo models of diabetic nephropathy, respectively. KLF6 overexpression increased Txnip expression and promoter activity, which was inhibited by concurrent exposure to PPAR-gamma agonists. In contrast, reduced expression of KLF6 by siRNA or exposure to PPAR-gamma agonists attenuated high glucose-induced Txnip expression and promoter activity. KLF6 Txnip promoter binding was decreased in KLF6-silenced cells, whereas PPAR-gamma agonists increased PPAR-gamma-Txnip promoter binding. Indeed, silencing of KLF6 increased PPAR-gamma expression, suggesting endogenous regulation of PPAR-gamma expression by KLF6. Moreover, renal KLF6 and Txnip expression increased in rats with diabetes mellitus and was inhibited by PPAR-gamma agonist treatment; however, KLF6 expression did not change in HK-2 cells exposed to PPAR-gamma agonists. Hence, Txnip expression and promoter activity are mediated via divergent effects of KLF6 and PPAR-gamma transcriptional regulation. PMID- 19808646 TI - {alpha}7 nicotinic acetylcholine receptor regulates airway epithelium differentiation by controlling basal cell proliferation. AB - Airway epithelial basal cells are known to be critical for regenerating injured epithelium and maintaining tissue homeostasis. Recent evidence suggests that the alpha7 nicotinic acetylcholine receptor (nAChR), which is highly permeable to Ca(2+), is involved in lung morphogenesis. Here, we have investigated the potential role of the alpha7 nAChR in the regulation of airway epithelial basal cell proliferation and the differentiation of the human airway epithelium. In vivo during fetal development and in vitro during the regeneration of the human airway epithelium, alpha7 nAChR expression coincides with epithelium differentiation. Inactivating alpha7 nAChR function in vitro increases cell proliferation during the initial steps of the epithelium regeneration, leading to epithelial alterations such as basal cell hyperplasia and squamous metaplasia, remodeling observed in many bronchopulmonary diseases. The regeneration of the airway epithelium after injury in alpha7(-/-) mice is delayed and characterized by a transient hyperplasia of basal cells. Moreover, 1-year-old alpha7(-/-) mice more frequently present basal cells hyperplasia. Modulating nAChR function or expression shows that only alpha7 nAChR, as opposed to heteropentameric alpha(x)beta(y) nAChRs, controls the proliferation of human airway epithelial basal cells. These findings suggest that alpha7 nAChR is a key regulator of the plasticity of the human airway epithelium by controlling basal cell proliferation and differentiation pathway and is involved in airway remodeling during bronchopulmonary diseases. PMID- 19808647 TI - Type I interferon modulates monocyte recruitment and maturation in chronic inflammation. AB - Chronic inflammation is characterized by continuous recruitment and activation of immune cells such as monocytes in response to a persistent stimulus. Production of proinflammatory mediators by monocytes leads to tissue damage and perpetuates the inflammatory response. However, the mechanism(s) responsible for the sustained influx of monocytes in chronic inflammation are not well defined. In chronic peritonitis induced by pristane, the persistent recruitment of Ly6C(hi) inflammatory monocytes into the peritoneum was abolished in type I interferon (IFN-I) receptor-deficient mice but was unaffected by the absence of IFN-gamma, tumor necrosis factor-alpha, interleukin-6, or interleukin-1. IFN-I signaling stimulated the production of chemokines (CCL2, CCL7, and CCL12) that recruited Ly6C(hi) monocytes via interactions with the chemokine receptor CCR2. Interestingly, after 2,6,10,14-tetramethylpentadecane treatment, the rapid turnover of inflammatory monocytes in the inflamed peritoneum was associated with a lack of differentiation into Ly6C(lo) monocytes/macrophages, a more mature subset with enhanced phagocytic capacity. In contrast, Ly6C(hi) monocytes differentiated normally into Ly6C(lo) cells in IFN-I receptor-deficient mice. The effects of IFN-I were specific for monocytes as granulocyte migration was unaffected in the absence of IFN-I signaling. Taken together, our findings reveal a novel role of IFN-I in promoting the recruitment of inflammatory monocytes via the chemokine receptor CCR2. Continuous monocyte recruitment and the lack of terminal differentiation induced by IFN-I may help sustain the chronic inflammatory response. PMID- 19808648 TI - Age-related changes in pericellular hyaluronan organization leads to impaired dermal fibroblast to myofibroblast differentiation. AB - We have previously demonstrated that transforming growth factor-beta1 (TGF-beta1) mediated fibroblast-myofibroblast differentiation is associated with accumulation of a hyaluronan (HA) pericellular coat. The current study demonstrates failure of fibroblast-myofibroblast differentiation associated with in vitro aging. This is associated with attenuation of numerous TGF-beta1-dependent responses, including HA synthesis and induction of the HA synthase enzyme HAS2 and the hyaladherin tumor necrosis factor-alpha-stimulated gene 6 (TSG-6), which led to an age related defect in pericellular HA coat assembly. Inhibition of HAS2-dependent HA synthesis by gene silencing, removal of the HA coat by hyaluronidase digestion, or gene silencing of TSG-6 or cell surface receptor CD44 led to abrogation of TGF beta1-dependent induction of alpha-smooth muscle actin in "young" cells. This result supports the importance of HAS2-dependent HA synthesis and the HA coat during phenotypic activation. Interleukin-1beta stimulation, however, failed to promote phenotypic conversion despite coat formation. A return to basal levels of HA synthesis in aged cells by HAS2 overexpression restored TGF-beta1-dependent induction of TSG-6 and pericellular HA coat assembly. However, this did not lead to the acquisition of a myofibroblast phenotype. Coordinated induction of HAS2 and TSG-6 facilitation of pericellular HA coat assembly is necessary for TGF beta1-dependent activation of fibroblasts, and both components of this response are impaired with in vitro aging. In conclusion, the HA pericellular coat is integral but not sufficient to correct for the age-dependent defect in phenotypic conversion. PMID- 19808649 TI - A mouse model of inducible liver injury caused by tet-on regulated urokinase for studies of hepatocyte transplantation. AB - Mouse models of liver injury provide useful tools for studying hepatocyte engraftment and proliferation. A representative model of liver injury is the albumin-urokinase (Alb-uPA) transgenic model, but neonatal lethality hampers its widespread application. To overcome this problem, we generated a transgenic mouse in which transcription of the reverse tetracycline transactivator was (rtTA) driven by the mouse albumin promoter, and backcrossed the rtTA mice onto severe combined immunodeficient (SCID)/bg mice to generate immunodeficient rtTA/SCID mice. We then produced recombinant adenoviruses Ad.TRE-uPA, in which the urokinase was located downstream of the tetracycline response element (TRE). The rtTA/SCID mouse hepatocytes were then infected with Ad.TRE-uPA to establish an inducible liver injury mouse model. In the presence of doxycycline, uPA was exclusively expressed in endogenous hepatocytes and caused extensive liver injury. Enhanced green fluorescent protein-labeled mouse hepatocytes selectively repopulated the rtTA/SCID mouse liver and replaced over 80% of the recipient liver mass after repeated administration of Ad.TRE-uPA. Compared with the original uPA mice, rtTA/SCID mice did not exhibit problems regarding breeding efficiency, and the time window for transplantation was flexible. In addition, we could control the extent of liver injury to facilitate transplantation surgery by regulating the dose of Ad.TRE-uPA. Our inducible mouse model will be convenient for studies of hepatocyte transplantation and hepatic regeneration, and this system will facilitate screening for potential genetic factors critical for engraftment and proliferation of hepatocytes in vivo. PMID- 19808650 TI - Protection against hepatocyte mitochondrial dysfunction delays fibrosis progression in mice. AB - Accumulating evidence indicates that oxidative stress is involved in the physiopathology of liver fibrogenesis. However, amid the global context of hepatic oxidative stress, the specific role of hepatocyte mitochondrial dysfunction in the fibrogenic process is still unknown. The aim of this study was to determine whether a targeted protection of hepatocytes against mitochondrial dysfunction could modulate fibrosis progression. We induced liver fibrogenesis by chronic carbon tetrachloride treatment (3 or 6 weeks of biweekly injections) in transgenic mice expressing Bcl-2 in their hepatocytes or in normal control mice. Analyses of mitochondrial DNA, respiratory chain complexes, and lipid peroxidation showed that Bcl-2 transgenic animals were protected against mitochondrial dysfunction and oxidative stress resulting from carbon tetrachloride injury. Picrosirius red staining, alpha-smooth muscle actin immunohistochemistry, and real-time PCR for transforming growth factor-beta and collagen alpha-I revealed that Bcl-2 transgenic mice presented reduced fibrosis at early stages of fibrogenesis. However, at later stages increased nonmitochondrial/nonhepatocytic oxidative stress eventually overcame the capacity of Bcl-2 overexpression to prevent the fibrotic process. In conclusion, we demonstrate for the first time that specific protection against hepatocyte mitochondrial dysfunction plays a preventive role in early stages of fibrogenesis, delaying its onset. However, with the persistence of the aggression, this protection is no longer sufficient to impede fibrosis progression. PMID- 19808651 TI - Interferon-{gamma} differentially affects Alzheimer's disease pathologies and induces neurogenesis in triple transgenic-AD mice. AB - Inflammatory processes, including the episodic and/ or chronic elaboration of cytokines, have been identified as playing key roles in a number of neurological disorders. Whether these activities impart a disease-resolving and/or contributory outcome depends at least in part on the disease context, stage of pathogenesis, and cellular milieu in which these factors are released. Interferon gamma (IFNgamma) is one such cytokine that produces pleiotropic effects in the brain. It is protective by ensuring maintenance of virus latency after infection, yet deleterious by recruiting and activating microglia that secrete potentially damaging factors at sites of brain injury. Using the triple-transgenic mouse model of Alzheimer's disease (3xTg-AD), which develops amyloid and tau pathologies in a pattern reminiscent of human Alzheimer's disease, we initiated chronic intrahippocampal expression of IFNgamma through delivery of a serotype-1 recombinant adeno-associated virus vector (rAAV1-IFNgamma). Ten months of IFNgamma expression led to an increase in microglial activation, steady-state levels of proinflammatory cytokine and chemokine transcripts, and severity of amyloid-related pathology. In contrast, these rAAV1-IFNgamma-treated 3xTg-AD mice also exhibited diminished phospho-tau pathology and evidence of increased neurogenesis. Overall, IFNgamma mediates what seem to be diametrically opposed functions in the setting of AD-related neurodegeneration. Gaining an understanding as to how these apparently divergent functions are interrelated and controlled could elucidate new therapeutic strategies designed to harness the neuroprotective activity of IFNgamma. PMID- 19808652 TI - Correlation of CXCL12 expression and FoxP3+ cell infiltration with human papillomavirus infection and clinicopathological progression of cervical cancer. AB - Human cervical cancer is an immunogenic tumor with a defined pattern of histopathological and clinical progression. Tumor-infiltrating T cells contribute to immune control of this tumor; however, cervical cancer dysregulates this immune response both through its association with human papillomavirus (HPV) infection and by producing cytokines and chemokines. Animal tumor models have revealed associations between overproduction of the chemokine stromal cell derived factor-1 (SDF-1 or CXCL12) and dysregulation of tumor-specific immunity. We therefore proposed that CXCL12 expression by cervical precancerous and cancerous lesions correlates with histopathological progression, loss of immune control of the tumor, and HPV infection. We found a significant association between cancer stage and CXCL12 expression for squamous and glandular lesions as well as with the HPV16+ (high-risk) status of the neoplastic lesions. Cancer progression was correlated with increasing levels of FoxP3 T-cell infiltration in the tumor. FoxP3 and CXCL12 expression significantly correlated for squamous and glandular neoplastic lesions. These observations were supported by enzyme-linked immunosorbent assay and Western blotting. In addition, we demonstrated CXCL12 expression by dyskaryotic cells in ThinPrep cervical smears. This study robustly links increased CXCL12 expression and FoxP3(+)-cell infiltration to HPV infection and progression of cervical cancer. It supports the detection of CXCL12 in cervical smears and biopsies as an additional biomarker for this disease. PMID- 19808653 TI - Heightened colon motor activity measured by a wireless capsule in patients with constipation: relation to colon transit and IBS. AB - Relationships of regional colonic motility to transit in health, constipation, and constipation-predominant irritable bowel syndrome (C-IBS) are poorly characterized. This study aimed to 1) characterize regional differences in colon pressure, 2) relate motor differences in constipation to colon transit, and 3) quantify the role of IBS in altered contractility with constipation. Colon pH and pressure were measured by wireless capsules in 53 healthy and 36 constipated subjects. Numbers of contractions >25 mmHg and areas under curves (AUC) were calculated for colon transit quartiles by time. Constipation was classified as normal transit (<59 h), moderate slow transit (STC) (59-100 h), and severe STC (>100 h). Twelve out of 36 constipated subjects had C-IBS; 24 had functional constipation. Numbers of contractions and AUCs increased from the first to the fourth quartile in health (P < 0.0001). Mean numbers of contractions in constipated subjects were similar to controls. Mean AUCs with normal transit (P = 0.01) and moderate STC (P = 0.004) but not severe STC (P = NS) were higher than healthy subjects. IBS was associated with greater mean numbers of contractions (P = 0.05) and AUCs (P = 0.0006) vs. controls independent of transit. Numbers of contractions increased from the first to fourth quartiles in moderate STC, C-IBS, and functional constipation; AUCs increased from the first to fourth quartiles in all groups (all P < 0.05). In conclusion, colon pressure activity is greater distally than proximally in health. Constipated patients with normal or moderately delayed transit show increased motor activity that is partly explained by IBS. These findings emphasize differential effects on transit and motility in different constipation subtypes. PMID- 19808654 TI - Pdx1 inactivation restricted to the intestinal epithelium in mice alters duodenal gene expression in enterocytes and enteroendocrine cells. AB - Null mutant mice lacking the transcription factor pancreatic and duodenal homeobox 1 (Pdx1) are apancreatic and survive only a few days after birth. The role of Pdx1 in regulating intestinal gene expression has therefore yet to be determined in viable mice with normal pancreatic development. We hypothesized that conditional inactivation of Pdx1 restricted to the intestinal epithelium would alter intestinal gene expression and cell differentiation. Pdx1(flox/flox);VilCre mice with intestine-specific Pdx1 inactivation were generated by crossing a transgenic mouse strain expressing Cre recombinase, driven by a mouse villin 1 gene promoter fragment, with a mutant mouse strain homozygous for loxP site-flanked Pdx1. Pdx1 protein is undetectable in all epithelial cells in the intestinal epithelium of Pdx1(flox/flox);VilCre mice. Goblet cell number and mRNA abundance for mucin 3 and mucin 13 genes in the proximal small intestine are comparable between Pdx1(flox/flox);VilCre and control mice. Similarly, Paneth cell number and expression of Paneth cell-related genes Defa1, Defcr-rs1, and Mmp7 in the proximal small intestine remain statistically unchanged by Pdx1 inactivation. Although the number of enteroendocrine cells expressing chromogranin A/B, gastric inhibitory polypeptide (Gip), or somatostatin (Sst) is unaffected in the Pdx1(flox/flox);VilCre mice, mRNA abundance for Gip and Sst is significantly reduced in the proximal small intestine. Conditional Pdx1 inactivation attenuates intestinal alkaline phosphatase (IAP) activity in the duodenal epithelium, consistent with an average 91% decrease in expression of the mouse enterocyte IAP gene, alkaline phosphatase 3 (a novel Pdx1 target candidate), in the proximal small intestine following Pdx1 inactivation. We conclude that Pdx1 is necessary for patterning appropriate gene expression in enterocytes and enteroendocrine cells of the proximal small intestine. PMID- 19808655 TI - Carbohydrate maldigestion induces necrotizing enterocolitis in preterm pigs. AB - Necrotizing enterocolitis (NEC) remains the most severe gastrointestinal disorder in preterm infants. It is associated with the initiation of enteral nutrition and may be related to immature carbohydrate digestive capacity. We tested the hypothesis that a formula containing maltodextrin vs. a formula containing lactose as the principal source of carbohydrate would predispose preterm pigs to a higher NEC incidence. Cesarean-derived preterm pigs were given total parenteral nutrition for 48 h followed by total enteral nutrition with a lactose-based (n = 11) or maltodextrin-based (n = 11) formula for 36 h. A higher incidence (91% vs. 27%) and severity (score of 3.3 vs. 1.8) of NEC were observed in the maltodextrin than in the lactose group. This higher incidence of NEC in the maltodextrin group was associated with significantly lower activities of lactase, maltase, and aminopeptidase; reduced villus height; transiently reduced in vivo aldohexose uptake; and reduced ex vivo aldohexose uptake capacity in the middle region of the small intestine. Bacterial diversity was low for both diets, but alterations in bacterial composition and luminal concentrations of short-chain fatty acids were observed in the maltodextrin group. In a second study, we quantified net portal absorption of aldohexoses (glucose and galactose) during acute jejunal infusion of a maltodextrin- or a lactose-based formula (n = 8) into preterm pigs. We found lower net portal aldohexose absorption (4% vs. 42%) and greater intestinal recovery of undigested carbohydrate (68% vs. 27%) in pigs acutely perfused with the maltodextrin-based formula than those perfused with the lactose based formula. The higher digestibility of the lactose than the maltodextrin in the formulas can be attributed to a 5- to 20-fold higher hydrolytic activity of tissue-specific lactase than maltases. We conclude that carbohydrate maldigestion is sufficient to increase the incidence and severity of NEC in preterm pigs. PMID- 19808656 TI - Effects of intravenous fructose on gastric emptying and antropyloroduodenal motility in healthy subjects. AB - Gastric emptying (GE) of glucose is regulated closely, not only as a result of inhibitory feedback arising from the small intestine, but also because of the resulting hyperglycemia. Fructose is used widely in the diabetic diet and is known to empty from the stomach slightly faster than glucose but substantially slower than water. The aims of this study were to determine whether intravenous (iv) fructose affects GE and antropyloroduodenal motility and how any effects compare to those induced by iv glucose. Six healthy males (age: 26.7 +/- 3.8 yr) underwent concurrent measurements of GE of a solid meal (100 g ground beef labeled with 20 MBq (99m)Tc-sulfur colloid) and antropyloroduodenal motility on three separate days in randomized order during iv infusion of either fructose (0.5 g/kg), glucose (0.5 g/kg), or isotonic saline for 20 min. GE (scintigraphy), antropyloroduodenal motility (manometry), and blood glucose (glucometer) were measured for 120 min. There was a rise in blood glucose (P < 0.001) after iv glucose (peak 16.4 +/- 0.6 mmol/l) but not after fructose or saline. Intravenous glucose and fructose both slowed GE substantially (P < 0.005 for both), without any significant difference between them. Between t = 0 and 30 min, the number of antral pressure waves was less after both glucose and fructose (P < 0.002 for both) than saline, and there were more isolated pyloric pressure waves during iv glucose (P = 0.003) compared with fructose and saline (P = NS for both) infusions. In conclusion, iv fructose slows GE and modulates gastric motility in healthy subjects, and the magnitude of slowing of GE is comparable to that induced by iv glucose. PMID- 19808657 TI - Insulin-like growth factor-binding protein-5 stimulates growth of human intestinal muscle cells by activation of G{alpha}i3. AB - In human intestinal smooth muscle cells, endogenous insulin-like growth factor-I (IGF-I) regulates growth and IGF-binding protein-5 (IGFBP-5) expression. The effects of IGF-I are facilitated by IGFBP-5. We previously showed that IGFBP-5 acts independently of IGF-I in human intestinal muscle to stimulate proliferation and upregulate IGF-I production by activation of Erk1/2 and p38 MAPK. Thus a positive feedback loop exists between IGF-I and IGFBP-5, whereby both stimulate muscle growth and production of the other factor. In Crohn's disease, IGF-I and IGFBP-5 expression are increased and contribute to stricture formation through this effect on muscle growth. To determine the signaling pathways coupling IGFBP 5 to MAPK activation and growth, smooth muscle cells were isolated from muscularis propria of human intestine and placed into primary culture. Erk1/2 and p38 MAPK activation and type I collagen production were measured by immunoblot. Proliferation was measured by [(3)H]thymidine incorporation. Activation of specific G proteins was measured by ELISA. AG1024, an IGF-I receptor tyrosine kinase inhibitor, was used to isolate the IGF-I-independent effects of IGFBP-5. IGFBP-5-induced phosphorylation of Erk1/2 and p38 MAPK and proliferation were abolished by pertussis toxin, implying the participation of Gi. IGFBP-5 specifically activated Gi3 but not other G proteins. Transfection of an inhibitory Galphai minigene specifically inhibited MAPK activation, proliferation, and both collagen-I and IGF-I production. Our results indicate that endogenous IGFBP-5 activates Gi3 and regulates smooth muscle growth, IGF-I production, and collagen production via the alpha-subunit of Gi3, independently of IGF-I, in normal human intestinal muscle cells. PMID- 19808658 TI - Bone marrow contributes to the population of pancreatic stellate cells in mice. AB - Activated pancreatic stellate cells (PSCs) play a pivotal role in the development of pancreatic fibrosis. The origin of activated PSCs has been thought to be transformation of quiescent PSCs residing locally in the pancreas. Recent studies have suggested that bone marrow (BM)-derived cells participate in regeneration processes in various organs. This study aimed to clarify the contribution of BM derived cells to the population of PSCs in mice. We transplanted BM cells from male enhanced green fluorescent protein transgenic mice into female C57BL/6 mice after lethal irradiation. Eight weeks after BM transplantation, chronic pancreatitis was induced by administration of six intra-abdominal injections of cerulein (50 microg/kg body wt) at 1-h intervals, 3 days per week, for the total of 6 wk. BM-derived cells were tracked by green fluorescent protein expression and in situ hybridization for the Y-chromosome. Eight weeks after BM transplantation, BM-derived cells accounted for 8.7% of the desmin (a marker of PSCs)-positive cells in the pancreas. We could isolate BM-derived cells, which contained lipid droplets and expressed desmin. They could be transformed to myofibroblast-like cells by culture in vitro, further supporting that BM contributed to the population of quiescent PSCs. After induction of pancreatic fibrosis, BM-derived cells accounted for 20.2% of alpha-smooth muscle actin positive activated PSCs. The contribution of BM-derived cells to pancreatic ductal cells (positive for cytokeratin-19) was rare and less than 1%. In conclusion, our results suggested that BM-derived cells contributed to the population of PSCs in mice. PMID- 19808659 TI - CFTR knockdown stimulates lipid synthesis and transport in intestinal Caco-2/15 cells. AB - Cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel highly expressed in epithelial cells of the gastrointestinal tract. Mutations in the CFTR gene cause cystic fibrosis (CF), a disease characterized by pancreatic insufficiency, fat malabsorption, and steatorrhea. Despite the administration of pancreatic enzymes to normalize malabsorption, CF patients still experienced lipid fecal loss, nutritional deficiencies, and abnormalities in serum lipid profile, suggesting the presence of intrinsic defects in the intestinal handling of nutrients. The objective of the present study was to assess the impact of CFTR gene knockdown on intracellular lipid metabolism of the intestinal Caco-2/15 cell line. Partial CFTR gene inactivation led to cellular lipid accretion of phospholipids, triglycerides, and cholesteryl esters. Likewise, secretion of these lipid fractions was significantly increased following CFTR gene manipulation. As expected from these findings, the output of triglyceride-rich lipoproteins showed the same increasing pattern. Investigation of the mechanisms underlying these changes revealed that CFTR knockdown resulted in raised levels of apolipoproteins in cells and media and microsomal transfer protein activity, two important factors for the efficient assembly and secretion of lipoproteins. Similarly, scrutiny of the enzymatic monoacylglycerol acyltransferase and diacylglycerol acyltransferase, which exhibit dynamic function in triacylglycerol resynthesis and chylomicron formation in enterocytes, revealed a significant augmentation in their activity. Conversely, cholesterol uptake mediated by Niemann-Pick C1 like 1, Scavenger Receptor Class B Type I, and ATP-binding cassette G8 remains unaffected by genetic modification of CFTR. Collectively, these results highlight the role played by CFTR in intestinal handling of lipids and may suggest that factors other than defective CFTR are responsible for the abnormal intracellular events leading to fat malabsorption in CF patients. PMID- 19808660 TI - Activation of adenosine low-affinity A3 receptors inhibits the enteric short interplexus neural circuit triggered by histamine. AB - We tested the novel hypothesis that endogenous adenosine (eADO) activates low affinity A3 receptors in a model of neurogenic diarrhea in the guinea pig colon. Dimaprit activation of H2 receptors was used to trigger a cyclic coordinated response of contraction and Cl(-) secretion. Contraction-relaxation was monitored by sonomicrometry (via intracrystal distance) simultaneously with short-circuit current (I(sc), Cl(-) secretion). The short interplexus reflex coordinated response was attenuated or abolished by antagonists at H2 (cimetidine), 5 hydroxytryptamine 4 receptor (RS39604), neurokinin-1 receptor (GR82334), or nicotinic (mecamylamine) receptors. The A1 agonist 2-chloro-N(6) cyclopentyladenosine (CCPA) abolished coordinated responses, and A1 antagonists could restore normal responses. A1-selective antagonists alone [8 cyclopentyltheophylline (CPT), 1,3-dipropyl-8-(2-amino-4-chlorophenyl)xanthine (PACPX), or 8-cyclopentyl-N(3)-[3-(4-(fluorosulfonyl)benzoyloxy)propyl]-xanthine (FSCPX)] caused a concentration-dependent augmentation of crypt cell secretion or contraction and acted at nanomolar concentrations. The A3 agonist N(6)-(3 iodobenzyl)-adenosine-5'-N-methyluronamide (IB-MECA) abolished coordinated responses and the A3 antagonist 3-ethyl-5-benzyl-2-methyl-4-phenylethynyl-6 phenyl-1,4-(+/-)-dihydropyridine-3,5-dicarboxylate (MRS1191) could restore and further augment responses. The IB-MECA effect was resistant to knockdown of adenosine A1 receptor with the irreversible antagonist FSCPX; the IC(50) for IB MECA was 0.8 microM. MRS1191 alone could augment or unmask coordinated responses to dimaprit, and IB-MECA suppressed them. MRS1191 augmented distension-evoked reflex I(sc) responses. Adenosine deaminase mimicked actions of adenosine receptor antagonists. A3 receptor immunoreactivity was differentially expressed in enteric neurons of different parts of colon. After tetrodotoxin, IB-MECA caused circular muscle relaxation. The data support the novel concept that eADO acts at low-affinity A3 receptors in addition to high-affinity A1 receptors to suppress coordinated responses triggered by immune-histamine H2 receptor activation. The short interplexus circuit activated by histamine involves adenosine, acetylcholine, substance P, and serotonin. We postulate that A3 receptor modulation may occur in gut inflammatory diseases or allergic responses involving mast cell and histamine release. PMID- 19808661 TI - Autotaxin/lysophospholipase D-mediated lysophosphatidic acid signaling is required to form distinctive large lysosomes in the visceral endoderm cells of the mouse yolk sac. AB - Autotaxin, a lysophospholipase D encoded by the Enpp2 gene, is an exoenzyme that produces lysophosphatidic acid in the extracellular space. Lysophosphatidic acid acts on specific G protein-coupled receptors, thereby regulating cell growth, migration, and survival. Previous studies have revealed that Enpp2(-/-) mouse embryos die at about embryonic day (E) 9.5 because of angiogenic defects in the yolk sac. However, what cellular defects occur in Enpp2(-/-) embryos and what intracellular signaling pathways are involved in the phenotype manifestation remain unknown. Here, we show that Enpp2 is required to form distinctive large lysosomes in the yolk sac visceral endoderm cells. From E7.5 to E9.5, Enpp2 mRNA is abundantly expressed in the visceral endoderm cells. In Enpp2(-/-) mouse embryos, lysosomes in the visceral endoderm cells are fragmented. By using a whole embryo culture system combined with specific pharmacological inhibitors for intracellular signaling molecules, we show that lysophosphatidic acid receptors and the Rho-Rho-associated coiled-coil containing protein kinase (ROCK)-LIM kinase pathway are required to form large lysosomes. In addition, electroporation of dominant negative forms of Rho, ROCK, or LIM kinase also leads to the size reduction of lysosomes in wild-type visceral endoderm cells. In Enpp2(-/-) visceral endoderm cells, the steady-state levels of cofilin phosphorylation and actin polymerization are reduced. In addition, perturbations of actin turnover dynamics by actin inhibitors cytochalasin B and jasplakinolide result in the defect in lysosome formation. These results suggest that constitutive activation of the Rho-ROCK-LIM kinase pathway by extracellular production of lysophosphatidic acid by the action of autotaxin is required to maintain the large size of lysosomes in visceral endoderm cells. PMID- 19808662 TI - Evidence that nucleosomes inhibit mismatch repair in eukaryotic cells. AB - The influence of chromatin structure on DNA metabolic processes, including DNA replication and repair, has been a matter of intensive studies in recent years. Although the human mismatch repair (MMR) reaction has been reconstituted using purified proteins, the influence of chromatin structure on human MMR is unknown. This study examines the interaction between human MutSalpha and a mismatch located within a nucleosome or between two nucleosomes. The results show that, whereas MutSalpha specifically recognizes both types of nucleosomal heteroduplexes, the protein bound the mismatch within a nucleosome with much lower efficiency than a naked heteroduplex or a heterology free of histone proteins but between two nucleosomes. Additionally, MutSalpha displays reduced ATPase- and ADP-binding activity when interacting with nucleosomal heteroduplexes. Interestingly, nucleosomes block ATP-induced MutSalpha sliding along the DNA helix when the mismatch is in between two nucleosomes. These findings suggest that nucleosomes may inhibit MMR in eukaryotic cells. The implications of these findings for our understanding of eukaryotic MMR are discussed. PMID- 19808663 TI - Nrf2-dependent and -independent responses to nitro-fatty acids in human endothelial cells: identification of heat shock response as the major pathway activated by nitro-oleic acid. AB - Electrophilic fatty acid derivatives, including nitrolinoleic acid and nitro oleic acid (OA-NO(2)), can mediate anti-inflammatory and pro-survival signaling reactions. The transcription factor Nrf2, activated by electrophilic fatty acids, suppresses redox-sensitive pro-inflammatory gene expression and protects against vascular endothelial oxidative injury. It was therefore postulated that activation of Nrf2 by OA-NO(2) accounts in part for its anti-inflammatory actions, motivating the characterization of Nrf2-dependent and -independent effects of OA-NO(2) on gene expression using genome-wide transcriptional profiling. Control and Nrf2-small interfering RNA-transfected human endothelial cells were treated with vehicle, oleic acid, or OA-NO(2), and differential gene expression profiles were determined. Although OA-NO(2) significantly induced the expression of Nrf2-dependent genes, including heme oxygenase-1 and glutamate cysteine ligase modifier subunit, the majority of OA-NO(2)-regulated genes were regulated by Nrf2-independent pathways. Moreover, gene set enrichment analysis revealed that the heat shock response is the major pathway activated by OA-NO(2), with robust induction of a number of heat shock genes regulated by the heat shock transcription factor. Inasmuch as the heat shock response mediates anti inflammatory and cytoprotective actions, this mechanism is proposed to contribute to the protective cell signaling functions of nitro-fatty acids and other electrophilic fatty acid derivatives. PMID- 19808664 TI - A double tyrosine motif in the cardiac sodium channel domain III-IV linker couples calcium-dependent calmodulin binding to inactivation gating. AB - Voltage-gated sodium channels maintain the electrical cadence and stability of neurons and muscle cells by selectively controlling the transmembrane passage of their namesake ion. The degree to which these channels contribute to cellular excitability can be managed therapeutically or fine-tuned by endogenous ligands. Intracellular calcium, for instance, modulates sodium channel inactivation, the process by which sodium conductance is negatively regulated. We explored the molecular basis for this effect by investigating the interaction between the ubiquitous calcium binding protein calmodulin (CaM) and the putative sodium channel inactivation gate composed of the cytosolic linker between homologous channel domains III and IV (DIII-IV). Experiments using isothermal titration calorimetry show that CaM binds to a novel double tyrosine motif in the center of the DIII-IV linker in a calcium-dependent manner, N-terminal to a region previously reported to be a CaM binding site. An alanine scan of aromatic residues in recombinant DIII-DIV linker peptides shows that whereas multiple side chains contribute to CaM binding, two tyrosines (Tyr(1494) and Tyr(1495)) play a crucial role in binding the CaM C-lobe. The functional relevance of these observations was then ascertained through electrophysiological measurement of sodium channel inactivation gating in the presence and absence of calcium. Experiments on patch-clamped transfected tsA201 cells show that only the Y1494A mutation of the five sites tested renders sodium channel steady-state inactivation insensitive to cytosolic calcium. The results demonstrate that calcium-dependent calmodulin binding to the sodium channel inactivation gate double tyrosine motif is required for calcium regulation of the cardiac sodium channel. PMID- 19808665 TI - Characterization of the structure and intermolecular interactions between the connexin40 and connexin43 carboxyl-terminal and cytoplasmic loop domains. AB - Gap junctions are intercellular channels that allow the passage of ions, small molecules, and second messengers that are essential for the coordination of cellular function. They are formed by two hemichannels, each constituted by the oligomerization of six connexins (Cx). Among the 21 different human Cx isoforms, studies have suggested that in the heart, Cx40 and Cx43 can oligomerize to form heteromeric hemichannels. The mechanism of heteromeric channel regulation has not been clearly defined. Tissue ischemia leads to intracellular acidification and closure of Cx43 and Cx40 homomeric channels. However, coexpression of Cx40 and Cx43 in Xenopus oocytes enhances the pH sensitivity of the channel. This phenomenon requires the carboxyl-terminal (CT) part of both connexins. In this study we used different biophysical methods to determine the structure of the Cx40CT and characterize the Cx40CT/Cx43CT interaction. Our results revealed that the Cx40CT is an intrinsically disordered protein similar to the Cx43CT and that the Cx40CT and Cx43CT can interact. Additionally, we have identified an interaction between the Cx40CT and the cytoplasmic loop of Cx40 as well as between the Cx40CT and the cytoplasmic loop of Cx43 (and vice versa). Our studies support the "particle-receptor" model for pH gating of Cx40 and Cx43 gap junction channels and suggest that interactions between cytoplasmic regulatory domains (both homo- and hetero-connexin) could be important for the regulation of heteromeric channels. PMID- 19808666 TI - Regulation of protein kinase A activity by p90 ribosomal S6 kinase 1. AB - Previously, we reported that the catalytic subunit of cAMP-dependent protein kinase (PKAc) binds to the active p90 ribosomal S6 kinase 1 (RSK1) (Chaturvedi, D., Poppleton, H. M., Stringfield, T., Barbier, A., and Patel, T. B. (2006) Mol. Cell. Biol. 26, 4586-4600). Herein, by overexpressing hemagglutinin-tagged RSK1 fragments in HeLa cells we have identified the region of RSK1 that is responsible for the interaction with PKAc. PKAc bound to the last 13 amino acids of RSK1, which overlaps the Erk1/2 docking site. This interaction between PKAc and RSK1 required the phosphorylation of Ser-732 in the C terminus of RSK1. Depending upon its phosphorylation status, RSK1 switched interactions between Erk1/2 and PKAc. In addition, a peptide corresponding to the last 13 amino acids of RSK1 with substitution of Ser-732 with Glu (peptide E), but not Ala (peptide A), decreased interactions between endogenous active RSK1 and PKAc. RSK1 attenuated the ability of cAMP to activate PKA in vitro and this modulation was abrogated by peptide E, but not by peptide A. Similarly, in intact cells, cAMP-mediated phosphorylation of Bcl-xL/Bcl-2-associated death promoter on Ser-115, the PKA site, was reduced when RSK1 was activated by epidermal growth factor, and this effect was blocked by peptide E, but not by peptide A. These findings demonstrate that interactions between endogenous RSK1 and PKAc in intact cells regulate the ability of cAMP to activate PKA and identify a novel mechanism by which PKA activity is regulated by the Erk1/2 pathway. PMID- 19808667 TI - A non-hydrolyzable ATP derivative generates a stable complex in a light-inducible two-component system. AB - Isothermal calorimetry (ITC) measurements yielded the binding constants during complex formation of light-inducible histidine kinases (HK) and their cognate CheY-type response regulators (RR). HK-RR interactions represent the core function of the bacterial two-component system, which is also present in many bacterial phytochromes. Here, we have studied the recombinant forms of phytochromes CphA and CphB from the cyanobacterium Tolypothrix PCC7601 and their cognate RRs RcpA and RcpB. The interaction between the two reaction partners (HK and RR) was studied in the presence and absence of ATP. A complex formation was observable in the presence of ATP, but specific interactions were only found when a non-hydrolyzable ATP derivative was added to the mixture. Also, the incubation of the HK domain alone (expressed as a recombinant protein) with the RR did not yield specific interactions, indicating that the HK domain is only active as a component of the full-length phytochrome. Considering also previous studies on the same proteins (Hubschmann, T., Jorissen, H. J. M. M., Borner, T., Gartner, W., and de Marsac, N. (2001) Eur. J. Biochem. 268, 3383-3389) we now conclude that the HK domains of these phytochromes are active only when the chromophore domain is in its Pr form. The formerly documented phosphate transfer between the HK domain and the RR takes place via a transiently formed protein-protein complex, which becomes detectable by ITC in the presence of a non-hydrolyzable ATP derivative. This finding is of interest also in relation to the function of some (blue light-sensitive) photoreceptors that carry the HK domain and the RR fused together in one single protein. PMID- 19808668 TI - Predictive computational models of substrate binding by a nucleoside transporter. AB - Transporters play a vital role in both the resistance mechanisms of existing drugs and effective targeting of their replacements. Melarsoprol and diamidine compounds similar to pentamidine and furamidine are primarily taken up by trypanosomes of the genus Trypanosoma brucei through the P2 aminopurine transporter. In standardized competition experiments with [(3)H]adenosine, P2 transporter inhibition constants (K(i)) have been determined for a diverse dataset of adenosine analogs, diamidines, Food and Drug Administration-approved compounds and analogs thereof, and custom-designed trypanocidal compounds. Computational biology has been employed to investigate compound structure diversity in relation to P2 transporter interaction. These explorations have led to models for inhibition predictions of known and novel compounds to obtain information about the molecular basis for P2 transporter inhibition. A common pharmacophore for P2 transporter inhibition has been identified along with other key structural characteristics. Our model provides insight into P2 transporter interactions with known compounds and contributes to strategies for the design of novel antiparasitic compounds. This approach offers a quantitative and predictive tool for molecular recognition by specific transporters without the need for structural or even primary sequence information of the transport protein. PMID- 19808669 TI - A role for the ATP7A copper-transporting ATPase in macrophage bactericidal activity. AB - Copper is an essential micronutrient that is necessary for healthy immune function. This requirement is underscored by an increased susceptibility to bacterial infection in copper-deficient animals; however, a molecular understanding of its importance in immune defense is unknown. In this study, we investigated the effect of proinflammatory agents on copper homeostasis in RAW264.7 macrophages. Interferon-gamma was found to increase expression of the high affinity copper importer, CTR1, and stimulate copper uptake. This was accompanied by copper-stimulated trafficking of the ATP7A copper exporter from the Golgi to vesicles that partially overlapped with phagosomal compartments. Silencing of ATP7A expression attenuated bacterial killing, suggesting a role for ATP7A-dependent copper transport in the bactericidal activity of macrophages. Significantly, a copper-sensitive mutant of Escherichia coli lacking the CopA copper-transporting ATPase was hypersensitive to killing by RAW264.7 macrophages, and this phenotype was dependent on ATP7A expression. Collectively, these data suggest that copper-transporting ATPases, CopA and ATP7A, in both bacteria and macrophage are unique determinants of bacteria survival and identify an unexpected role for copper at the host-pathogen interface. PMID- 19808670 TI - A promiscuous conformational switch in the secondary multidrug transporter MdfA. AB - Multidrug (Mdr) transporters are membrane proteins that actively export structurally dissimilar drugs from the cell, thereby rendering the cell resistant to toxic compounds. Similar to substrate-specific transporters, Mdr transporters also undergo substrate-induced conformational changes. However, the mechanism by which a variety of dissimilar substrates are able to induce similar transport compatible conformational responses in a single transporter remains unclear. To address this major aspect of Mdr transport, we studied the conformational behavior of the Escherichia coli Mdr transporter MdfA. Our results show that indeed, different substrates induce similar conformational changes in the transporter. Intriguingly, in addition, we observed that compounds other than substrates are able to confer similar conformational changes when covalently attached at the putative Mdr recognition pocket of MdfA. Taken together, the results suggest that the Mdr-binding pocket of MdfA is conformationally sensitive. We speculate that the same conformational switch that usually drives active transport is triggered promiscuously by merely occupying the Mdr-binding site. PMID- 19808671 TI - Identification of a heterogeneous nuclear ribonucleoprotein-recognition region in the HIV Rev protein. AB - The Rev protein is a key regulator of human immunodeficiency virus type 1 (HIV-1) gene expression. Rev is primarily known as an adaptor protein for nuclear export of HIV RNAs. However, Rev also contributes to numerous other processes by less well known mechanisms. Understanding the functional nature of Rev requires extensive knowledge of its cellular interaction partners. Here we demonstrate that Rev interacts with members of a large family of multifunctional host cell factors called hnRNPs. Rev employs amino acids 9-14 for specific binding to the heterogeneous nuclear ribonucleoproteins (hnRNP) A1, Q, K, R, and U. In addition, Rev interacts with hnRNP E1 and E2 by a different mechanism. The set of hnRNPs recognized by the N terminus of Rev feature RGG boxes. Exemplary testing of hnRNP A1 revealed a critical role of arginine residues within the RGG box for interaction with Rev. Finally, we demonstrate that expression levels of hnRNP A1, Q, K, R, and U influence HIV-1 production by persistently infected astrocytes, linking these hnRNPs to HIV replication. The novel interaction of HIV-1 Rev with functionally diverse hnRNPs lends further support to the idea that Rev is a multifunctional protein and may be involved in coupling HIV replication to diverse cellular processes and promoting virus-host cell interactions. PMID- 19808672 TI - Multimerization and H3K9me3 binding are required for CDYL1b heterochromatin association. AB - Proteins containing defined recognition modules mediate readout and translation of histone modifications. These factors are thought to initiate downstream signaling events regulating chromatin structure and function. We identified CDYL1 as an interaction partner of histone H3 trimethylated on lysine 9 (H3K9me3). CDYL1 belongs to a family of chromodomain factors found in vertebrates. We show that three different splicing variants of CDYL1, a, b, and c, are differentially expressed in various tissues with CDYL1b being the most abundant variant. Although all three splicing variants share a common C-terminal enoyl-CoA hydratase-like domain, only CDYL1b contains a functional chromodomain implicated in H3K9me3 binding. A splicing event introducing an N-terminal extension right at the beginning of the chromodomain of CDYL1a inactivates its chromodomain. CDYL1c does not contain a chromodomain at all. Although CDYL1b displays binding affinity to methyl-lysine residues in different sequence context similar to chromodomains in other chromatin factors, we demonstrate that the CDYL1b chromodomain/H3K9me3 interaction is necessary but not sufficient for association of the factor with heterochromatin. Indeed, multimerization of the protein via the enoyl-CoA hydratase-like domain is essential for H3K9me3 chromatin binding in vitro and heterochromatin localization in vivo. In agreement, overexpression of CDYL1c that can multimerize, but does not interact with H3K9me3 can displace CDYL1b from heterochromatin. Our results imply that multimeric binding to H3K9me3 by CDYL1b homomeric complexes is essential for efficient chromatin targeting. We suggest that similar multivalent binding stably anchors other histone modification binding factors on their target chromatin regions. PMID- 19808673 TI - Differential effects of lysophosphatidic acid and phosphatidylinositol 4,5 bisphosphate on actin dynamics by direct association with the actin-binding protein villin. AB - We have previously reported that the epithelial cell-specific actin-binding protein villin directly associates with phosphatidylinositol 4,5-bisphosphate (PIP(2)) through three binding sites that overlap with actin-binding sites in villin. As a result, association of villin with PIP(2) inhibits actin depolymerization and enhances actin cross-linking by villin. In this study, we demonstrate that these three PIP(2)-binding sites also bind the more hydrophilic phospholipid, lysophosphatidic acid (LPA) but with a higher affinity than PIP(2) (dissociation constant (K(d)) of 22 mum versus 39.5 mum for PIP(2)). More interestingly, unlike PIP(2), the association of villin with LPA inhibits all actin regulatory functions of villin. In addition, unlike PIP(2), LPA dramatically stimulates the tyrosine phosphorylation of villin by c-Src kinase. These studies suggest that in cells, selective interaction of villin with either PIP(2) or LPA could have dramatically different outcomes on actin reorganization as well as phospholipid-regulated cell signaling. These studies provide a novel regulatory mechanism for phospholipid-induced changes in the microfilament structure and cell function and suggest that LPA could be an intracellular regulator of the actin cytoskeleton. PMID- 19808674 TI - Mechanism for amyloid precursor-like protein 2 enhancement of major histocompatibility complex class I molecule degradation. AB - Earlier studies have demonstrated interaction of the murine major histocompatibility complex (MHC) class I molecule K(d) with amyloid precursor like protein 2 (APLP2), a ubiquitously expressed member of the amyloid precursor protein family. Our current findings indicate that APLP2 is internalized in a clathrin-dependent manner, as shown by utilization of inhibitors of the clathrin pathway. Furthermore, we demonstrated that APLP2 and K(d) bind at the cell surface and are internalized together. The APLP2 cytoplasmic tail contains two overlapping consensus motifs for binding to the adaptor protein-2 complex, and mutation of a tyrosine shared by both motifs severely impaired APLP2 internalization and ability to promote K(d) endocytosis. Upon increased expression of wild type APLP2, K(d) molecules were predominantly directed to the lysosomes rather than recycled to the plasma membrane. These findings suggest a model in which APLP2 binds K(d) at the plasma membrane, facilitates uptake of K(d) in a clathrin-dependent manner, and routes the endocytosed K(d) to the lysosomal degradation pathway. Thus, APLP2 has a multistep trafficking function that influences the expression of major histocompatibility complex class I molecules at the plasma membrane. PMID- 19808675 TI - Hepatitis C virus proteins induce lipogenesis and defective triglyceride secretion in transgenic mice. AB - Chronic hepatitis C virus (HCV) infection is associated with altered lipid metabolism and hepatocellular steatosis. Virus-induced steatosis is a cytopathic effect of HCV replication. The goal of this study was to examine the mechanisms underlying HCV-induced lipid metabolic defects in a transgenic mouse model expressing the full HCV protein repertoire at levels corresponding to natural human infection. In this model, expression of the HCV full-length open reading frame was associated with hepatocellular steatosis and reduced plasma triglyceride levels. Triglyceride secretion was impaired, whereas lipogenesis was activated. Increased lipogenic enzyme transcription was observed, resulting from maturational activation and nuclear translocation of sterol regulatory element binding protein 1c (SREBP1c). However, endoplasmic reticulum (ER) stress markers were expressed at similar levels in both HCV transgenic mice and their wild type counterparts, suggesting that SREBP1c proteolytic cleavage in the presence of HCV proteins was independent of ER stress. In conclusion, transgenic mice expressing the HCV full-length polyprotein at low levels have decreased plasma triglyceride levels and develop hepatocellular steatosis in the same way as HCV-infected patients. In these mice, SREBP1c activation by one or several HCV proteins induces de novo triglyceride synthesis via the lipogenic pathway, in a manner independent of ER stress, whereas triglyceride secretion is simultaneously reduced. PMID- 19808676 TI - The target of the NSD family of histone lysine methyltransferases depends on the nature of the substrate. AB - The NSD (nuclear receptor SET domain-containing) family of histone lysine methyltransferases is a critical participant in chromatin integrity as evidenced by the number of human diseases associated with the aberrant expression of its family members. Yet, the specific targets of these enzymes are not clear, with marked discrepancies being reported in the literature. We demonstrate that NSD2 can exhibit disparate target preferences based on the nature of the substrate provided. The NSD2 complex purified from human cells and recombinant NSD2 both exhibit specific targeting of histone H3 lysine 36 (H3K36) when provided with nucleosome substrates, but histone H4 lysine 44 is the primary target in the case of octamer substrates, irrespective of the histones being native or recombinant. This disparity is negated when NSD2 is presented with octamer targets in conjunction with short single- or double-stranded DNA. Although the octamers cannot form nucleosomes, the target is nonetheless nucleosome-specific as is the product, dimethylated H3K36. This study clarifies in part the previous discrepancies reported with respect to NSD targets. We propose that DNA acts as an allosteric effector of NSD2 such that H3K36 becomes the preferred target. PMID- 19808677 TI - Fibril fragmentation enhances amyloid cytotoxicity. AB - Fibrils associated with amyloid disease are molecular assemblies of key biological importance, yet how cells respond to the presence of amyloid remains unclear. Cellular responses may not only depend on the chemical composition or molecular properties of the amyloid fibrils, but their physical attributes such as length, width, or surface area may also play important roles. Here, we report a systematic investigation of the effect of fragmentation on the structural and biological properties of amyloid fibrils. In addition to the expected relationship between fragmentation and the ability to seed, we show a striking finding that fibril length correlates with the ability to disrupt membranes and to reduce cell viability. Thus, despite otherwise unchanged molecular architecture, shorter fibrillar samples show enhanced cytotoxic potential than their longer counterparts. The results highlight the importance of fibril length in amyloid disease, with fragmentation not only providing a mechanism by which fibril load can be rapidly increased but also creating fibrillar species of different dimensions that can endow new or enhanced biological properties such as amyloid cytotoxicity. PMID- 19808678 TI - Protective role of Cys-178 against the inactivation and oligomerization of human insulin-degrading enzyme by oxidation and nitrosylation. AB - Insulin-degrading enzyme (IDE), a 110-kDa metalloendopeptidase, hydrolyzes several physiologically relevant peptides, including insulin and amyloid-beta (Abeta). Human IDE has 13 cysteines and is inhibited by hydrogen peroxide and S nitrosoglutathione (GSNO), donors of reactive oxygen and nitrogen species, respectively. Here, we report that the oxidative burst of BV-2 microglial cells leads to oxidation or nitrosylation of secreted IDE, leading to the reduced activity. Hydrogen peroxide and GSNO treatment of IDE reduces the V(max) for Abeta degradation, increases IDE oligomerization, and decreases IDE thermostability. Additionally, this inhibitory response of IDE is substrate dependent, biphasic for Abeta degradation but monophasic for a shorter bradykinin mimetic substrate. Our mutational analysis of IDE and peptide mass fingerprinting of GSNO-treated IDE using Fourier transform-ion cyclotron resonance mass spectrometer reveal a surprising interplay of Cys-178 with Cys-110 and Cys-819 for catalytic activity and with Cys-789 and Cys-966 for oligomerization. Cys-110 is near the zinc-binding catalytic center and is normally buried. The oxidation and nitrosylation of Cys-819 allow Cys-110 to be oxidized or nitrosylated, leading to complete inactivation of IDE. Cys-789 is spatially adjacent to Cys 966, and their nitrosylation and oxidation together trigger the oligomerization and inhibition of IDE. Interestingly, the Cys-178 modification buffers the inhibition caused by Cys-819 modification and prevents the oxidation or nitrosylation of Cys-110. The Cys-178 modification can also prevent the oligomerization-mediated inhibition. Thus, IDE can be intricately regulated by reactive oxygen or nitrogen species. The structure of IDE reveals the molecular basis for the long distance interactions of these cysteines and how they regulate IDE function. PMID- 19808679 TI - Cwp84, a surface-associated cysteine protease, plays a role in the maturation of the surface layer of Clostridium difficile. AB - Clostridium difficile is a major and growing problem as a hospital-associated infection that can cause severe, recurrent diarrhea. The mechanism by which the bacterium colonizes the gut during infection is poorly understood but undoubtedly involves protein components within the surface layer (S-layer), which play a role in adhesion. In C. difficile, the S-layer is composed of two principal components, the high and low molecular weight S-layer proteins, which are formed from the post-translational cleavage of a single precursor, SlpA. In the present study, we demonstrate that a recently characterized cysteine protease, Cwp84 plays a role in maturation of SlpA. Using a gene knock-out approach, we show that inactivation of the Cwp84 gene in C. difficile 630DeltaErm results in a bacterial phenotype in which only immature, single chain SlpA comprises the S-layer. The Cwp84 knock-out mutants (CDDeltaCwp84) displayed significantly different colony morphology compared with the wild-type strain and grew more slowly in liquid medium. SlpA extracted from CDDeltaCwp84 was readily cleaved into its mature subunits by trypsin treatment. Addition of trypsin to the growth medium also cleaved SlpA on CDDeltaCwp84 and increased the growth rate of the bacterium in a dose-dependent manner. Using the hamster model for C. difficile infection, CDDeltaCwp84 was found to be competent at causing disease with a similar pathology to the wild-type strain. The data show that whereas Cwp84 plays a role in the cleavage of SlpA, it is not an essential virulence factor and that bacteria expressing immature SlpA are able to cause disease. PMID- 19808680 TI - Domain II loop 3 of Bacillus thuringiensis Cry1Ab toxin is involved in a "ping pong" binding mechanism with Manduca sexta aminopeptidase-N and cadherin receptors. AB - Bacillus thuringiensis Cry toxins are used worldwide as insecticides in agriculture, in forestry, and in the control of disease transmission vectors. In the lepidopteran Manduca sexta, cadherin (Bt-R(1)) and aminopeptidase-N (APN) function as Cry1A toxin receptors. The interaction with Bt-R(1) promotes cleavage of the amino-terminal end, including helix alpha-1 and formation of prepore oligomer that binds to APN, leading to membrane insertion and pore formation. Loops of domain II of Cry1Ab toxin are involved in receptor interaction. Here we show that Cry1Ab mutants located in domain II loop 3 are affected in binding to both receptors and toxicity against Manduca sexta larvae. Interaction with both receptors depends on the oligomeric state of the toxin. Monomers of loop 3 mutants were affected in binding to APN and to a cadherin fragment corresponding to cadherin repeat 12 but not with a fragment comprising cadherin repeats 7-12. In contrast, the oligomers of loop 3 mutants were affected in binding to both Bt R(1) fragments but not to APN. Toxicity assays showed that either monomeric or oligomeric structures of Cry1Ab loop 3 mutations were severely affected in insecticidal activity. These data suggest that loop 3 is differentially involved in the binding with both receptor molecules, depending on the oligomeric state of the toxin and also that possibly a "ping pong" binding mechanism with both receptors is involved in toxin action. PMID- 19808681 TI - Triple N-glycosylation in the long S5-P loop regulates the activation and trafficking of the Kv12.2 potassium channel. AB - Mammalian voltage-dependent potassium (Kv) channels regulate the excitability of nerve and muscle cells. Kv12.2 features the longest S5-P loop among all known mammalian Kv channels with the most N-linked glycosylation sites (three sites). Despite its unique structural features, Kv12.2 is not well characterized. Because glycosylation plays important roles in the folding, trafficking, and function of various Kv channels, we focused on the N-glycosylation of Kv12.2. We show that Kv12.2 is N-glycosylated in Chinese hamster ovary (CHO) cells and in cultured neurons as well as in the mouse brain. As an effect of N-glycosylation on the function of Kv12.2, we demonstrate that removal of sugar chains causes a depolarizing shift in the steady-state activation without a significant reduction in current amplitude. Unlike the previously reported shift for Shaker-type Kv channels, this shift does not appear to be due to negatively charged sialic acid residues in the sugar chains. We next examined the trafficking in CHO cells to address whether the unglycosylated Kv12.2 channels are utilized in vivo. Although double mutants, retaining only one glycosylation site, are trafficked to the surface of CHO cells irrespective of the position of the glycosylated site, unglycosylated channels are not trafficked to the cell surface. Furthermore, we could not detect unglycosylated channels in the mouse brain. Our data suggest that only glycosylated Kv12.2 channels show proper voltage dependence and are utilized in vivo. PMID- 19808682 TI - 5-Nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) stimulates cellular ATP release through exocytosis of ATP-enriched vesicles. AB - Cells release ATP in response to physiologic stimuli. Extracellular ATP regulates a broad range of important cellular functions by activation of the purinergic receptors in the plasma membrane. The purpose of these studies was to assess the role of vesicular exocytosis in cellular ATP release. FM1-43 fluorescence was used to measure exocytosis and bioluminescence to measure ATP release in HTC rat hepatoma and Mz-Cha-1 human cholangiocarcinoma cells. Exposure to a Cl(-) channel inhibitor 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) (50-300 microM) stimulated a 5-100-fold increase in extracellular ATP levels within minutes of the exposure. This rapid response was not a result of changes in cell viability or Cl(-) channel activity. NPPB also potently stimulated ATP release in HEK293 cells and HEK293 cells expressing a rat P2X7 receptor indicating that P2X7 receptors are not involved in stimulation of ATP release by NPPB. In all cells studied, NPPB rapidly stimulated vesicular exocytosis that persisted many minutes after the exposure. The kinetics of NPPB-evoked exocytosis and ATP release were similar. Furthermore, the magnitudes of NPPB-evoked exocytosis and ATP release were correlated (correlation coefficient 0.77), indicating that NPPB may stimulate exocytosis of a pool of ATP-enriched vesicles. These findings provide further support for the concept that vesicular exocytosis plays an important role in cellular ATP release and suggest that NPPB can be used as a biochemical tool to specifically stimulate ATP release through exocytic mechanisms. PMID- 19808683 TI - Genetic evidence that an endosymbiont-derived endoplasmic reticulum-associated protein degradation (ERAD) system functions in import of apicoplast proteins. AB - Most apicomplexan parasites harbor a relict chloroplast, the apicoplast, that is critical for their survival. Whereas the apicoplast maintains a small genome, the bulk of its proteins are nuclear encoded and imported into the organelle. Several models have been proposed to explain how proteins might cross the four membranes that surround the apicoplast; however, experimental data discriminating these models are largely missing. Here we present genetic evidence that apicoplast protein import depends on elements derived from the ER-associated protein degradation (ERAD) system of the endosymbiont. We identified two sets of ERAD components in Toxoplasma gondii, one associated with the ER and cytoplasm and one localized to the membranes of the apicoplast. We engineered a conditional null mutant in apicoplast Der1, the putative pore of the apicoplast ERAD complex, and found that loss of Der1(Ap) results in loss of apicoplast protein import and subsequent death of the parasite. PMID- 19808684 TI - A combinatorial approach for the design of complementarity-determining region derived peptidomimetics with in vitro anti-tumoral activity. AB - The great success of therapeutic monoclonal antibodies has fueled research toward mimicry of their binding sites and the development of new strategies for peptide based mimetics production. Here, we describe a new combinatorial approach for the production of peptidomimetics using the complementarity-determining regions (CDRs) from gastrin17 (pyroEGPWLEEEEEAYGWMDF-NH(2)) antibodies as starting material for cyclic peptide synthesis in a microarray format. Gastrin17 is a trophic factor in gastrointestinal tumors, including pancreatic cancer, which makes it an interesting target for development of therapeutic antibodies. Screening of microarrays containing bicyclic peptidomimetics identified a high number of gastrin binders. A strong correlation was observed between gastrin binding and overall charge of the peptidomimetic. Most of the best gastrin binders proceeded from CDRs containing charged residues. In contrast, CDRs from high affinity antibodies containing mostly neutral residues failed to yield good binders. Our experiments revealed essential differences in the mode of antigen binding between CDR-derived peptidomimetics (K(d) values in micromolar range) and the parental monoclonal antibodies (K(d) values in nanomolar range). However, chemically derived peptidomimetics from gastrin binders were very effective in gastrin neutralization studies using cell-based assays, yielding a neutralizing activity in pancreatic tumoral cell lines comparable with that of gastrin specific monoclonal antibodies. These data support the use of combinatorial CDR peptide microarrays as a tool for the development of a new generation of chemically synthesized cyclic peptidomimetics with functional activity. PMID- 19808685 TI - Purification and reconstitution of the antigen transport complex TAP: a prerequisite for determination of peptide stoichiometry and ATP hydrolysis. AB - The transporter associated with antigen processing (TAP) is an essential machine of the adaptive immune system that translocates antigenic peptides from the cytosol into the endoplasmic reticulum lumen for loading of major histocompatibility class I molecules. To examine this ABC transport complex in mechanistic detail, we have established, after extensive screening and optimization, the solubilization, purification, and reconstitution for TAP to preserve its function in each step. This allowed us to determine the substrate binding stoichiometry of the TAP complex by fluorescence cross-correlation spectroscopy. In addition, the TAP complex shows strict coupling between peptide binding and ATP hydrolysis, revealing no basal ATPase activity in the absence of peptides. These results represent an optimal starting point for detailed mechanistic studies of the transport cycle of TAP by single molecule experiments to analyze single steps of peptide translocation and the stoichiometry between peptide transport and ATP hydrolysis. PMID- 19808693 TI - Longitudinal examination with shoulder ultrasound of patients with polymyalgia rheumatica. AB - OBJECTIVE: To determine if ultrasonography (US) and power Doppler (PD) may be useful in identifying polymyalgia rheumatica (PMR) patients with relapsing disease. METHODS: For a mean of 41 months, 57 consecutive untreated patients with PMR were prospectively assessed for relapses/recurrences. This cohort represented all the patients diagnosed over a 18-month period in one Italian secondary referral centre. Clinical signs and symptoms as well as ESR and CRP were evaluated. US examination of the shoulders was performed in all 57 patients at diagnosis and after the onset of prednisone treatment (mean 24 +/- 3 weeks). Power Doppler ultrasonography (PDUS) was performed in 24 patients. Shoulder sonograms were obtained according to standardized techniques. RESULTS: Prednisone therapy significantly reduced the frequency and the degree of subacromial/subdeltoid bursitis, long head biceps tenosynovitis and glenohumeral synovitis. At diagnosis, a positive PD signal was observed more frequently in the subacromial/subdeltoid bursae (33%). Prednisone therapy significantly reduced the frequency of patients with positive PD signal. Of the 44 patients in remission or with low disease activity at the time of the second US, 26 (59%) still had evidence of persistent inflammatory lesions. There was no association between the persistence of inflammation at US and relapses/recurrences; in contrast, a positive PD signal at diagnosis was significantly associated with the occurrence of relapses/recurrences at follow-up. CONCLUSION: Subclinical inflammation detected by US persists in most PMR patients despite glucocorticoid treatment. PDUS may be useful to detect at diagnosis the patients with most active inflammation who have a higher risk of relapses/recurrences. PMID- 19808686 TI - Phosphatase and tensin homologue on chromosome 10 (PTEN) directs prostaglandin E2 mediated fibroblast responses via regulation of E prostanoid 2 receptor expression. AB - Prostaglandin E(2) (PGE(2)) is an arachidonic acid metabolite that counters transforming growth factor-beta-induced fibroblast activation via E prostanoid 2 (EP2) receptor binding. Phosphatase and tensin homologue on chromosome 10 (PTEN) is a lipid phosphatase that, by antagonizing the phosphoinositol 3-kinase (PI3K) pathway, also inhibits fibroblast activation. Here, we show that PTEN directly regulates PGE(2) inhibition of fibroblast activation by augmenting EP2 receptor expression. The increase in collagen production and alpha-smooth muscle actin expression observed in fibroblasts in which PTEN is deficient was resistant to the usual suppressive effects of PGE(2). This was due to marked down-regulation of EP2, a G(s) protein-coupled receptor (GPCR) that mediates the inhibitory actions of this prostanoid via cAMP. pten(-/-) or PTEN-inhibited fibroblasts in which the PI3K pathway was blocked demonstrated a restoration of EP2 receptor expression, due to augmented gene transcription and mRNA instability. Importantly, restoration of the balance between PI3K and PTEN reestablished the inhibitory effect of PGE(2) on fibroblast activation. No such influence of PTEN was observed on alternative E prostanoid GPCRs. Moreover, our studies identified a positive feedback loop in which cAMP signaling enhanced EP2 receptor expression, independent of PTEN. Therefore, our findings indicate that PTEN regulates the antifibrotic effects of PGE(2) by a specific and permissive effect on EP2 receptor expression. Further, our data imply that cAMP signaling circumvents EP2 down-regulation in pten-deficient cells to restore EP2 receptor expression. This is the first description, to our knowledge, of PI3K/PTEN balance directing GPCR expression, and provides a novel mechanism for cellular effects of PTEN. PMID- 19808694 TI - Clinical usefulness of anti-RNA polymerase III antibody measurement by enzyme linked immunosorbent assay. AB - OBJECTIVE: To evaluate the clinical usefulness of measuring anti-RNA polymerase (RNAP) III antibody with a commercially available ELISA in Japanese patients with SSc. METHODS: This multicentre study involved 354 patients with SSc, 245 with non SSc CTDs and 102 healthy controls. ELISAs were used to detect anti-RNAP III antibody, anti-topo I antibody and ACA. The presence of anti-RNAP III antibody in selected serum samples was confirmed by immunoprecipitation (IP) assay. RESULTS: By ELISA, anti-RNAP III antibody was detected in 38 (10.7%) patients with SSc, 3 (1.2%) with non-SSc CTD and no healthy controls. The clinical specificity for SSc was excellent (98.8%), although a small number of false positives occurred. The sensitivity of the anti-topo I and ACA ELISAs for SSc was 59.9%, which increased to 68.2% without a reduction in specificity when the anti-RNAP III measurement was added. Clinical features associated with positivity for the anti-RNAP III antibody include dcSSc, a high total skin score and a trend towards high prevalence of renal crisis, consistent with previous studies that used an IP assay. Furthermore, on clinical severity scales, SSc patients with anti-RNAP III antibody scored highest for skin and renal involvement among patients subgrouped by the presence of individual SSc-related antibodies. CONCLUSIONS: The measurement of anti-RNAP III antibody by ELISA is useful in routine clinical practice, because it helps diagnose SSc and identify a disease subset with severe skin and renal involvement. PMID- 19808695 TI - The modern management of gout. AB - Gout is an inflammatory arthritis characterized by self-limiting but excruciatingly painful acute attacks. These are a consequence of monosodium urate (MSU) crystals being deposited within articular or periarticular tissue. Chronic tophaceous gout can develop after years of acute intermittent gout. Recent discoveries, including the role of the inflammasome and intracellular events demonstrating that pro-inflammatory cytokines, IL-1 beta, -8 and TNF-alpha, promote neutrophil influx. Also, genetic advances with the identification of the URAT-1 transporter and genetic variation in SLC 2A9 as a key regulator of urate homoeostasis, have given us deeper understanding of the pathophysiology of gout, and also allow for more targeted treatments. Hopefully, new and emerging therapeutic options will reduce treatment-resistant gout in patients who are unresponsive or unable to take traditional urate lowering therapy. The development of new therapies may also increase patient numbers being treated in the specialist setting, which may have several secondary benefits. PMID- 19808696 TI - Visualizing the von Willebrand factor/glycoprotein Ib-IX axis with a platelet type von Willebrand disease mutation. AB - Platelet-type von Willebrand disease (PT-VWD) is a bleeding disorder of the platelet glycoprotein Ib-IX/von Willebrand factor (VWF) axis caused by mutations in the glycoprotein Ib-IX receptor that lead to an increased affinity with VWF. In this report, platelets from a mouse expressing a mutation associated with PT VWD have been visualized using state-of-the art image collection and processing. Confocal analysis revealed that VWF bound to the surface of single platelets and bridging micro-aggregates of platelets. Surface-bound VWF appears as a large, linear structure on the surface of 50% of the PT-VWD platelets. In vivo thrombus formation after chemical injury to the carotid artery revealed a severe impairment to occlusion as a consequence of the PT-VWD mutation. In vitro stimulation of PT-VWD platelets with adenosine diphosphate or thrombin demonstrates a significant block in their ability to bind fibrinogen. The impairment of in vivo thrombus formation and in vitro fibrinogen binding are more significant than might be expected from the observed platelet binding to VWF polymers over a small portion of the plasma membrane. Visualization of the receptor/ligand interaction and characterization of a severe antithrombotic phenotype provide a new understanding on the molecular basis of bleeding associated with the PT-VWD phenotype. PMID- 19808697 TI - AML1/RUNX1 mutations in 470 adult patients with de novo acute myeloid leukemia: prognostic implication and interaction with other gene alterations. AB - Somatic mutation of the AML1/RUNX1(RUNX1) gene is seen in acute myeloid leukemia (AML) M0 subtype and in AML transformed from myelodysplastic syndrome, but the impact of this gene mutation on survival in AML patients remains unclear. In this study, we sought to determine the clinical implications of RUNX1 mutations in 470 adult patients with de novo non-M3 AML. Sixty-three distinct RUNX1 mutations were identified in 62 persons (13.2%); 32 were in N-terminal and 31, C-terminal. The RUNX1 mutation was closely associated with male sex, older age, lower lactic dehydrogenase value, French-American-British M0/M1 subtypes, and expression of HLA-DR and CD34, but inversely correlated with CD33, CD15, CD19, and CD56 expression. Furthermore, the mutation was positively associated with MLL/PTD but negatively associated with CEBPA and NPM1 mutations. AML patients with RUNX1 mutations had a significantly lower complete remission rate and shorter disease free and overall survival than those without the mutation. Multivariate analysis demonstrated that RUNX1 mutation was an independent poor prognostic factor for overall survival. Sequential analysis in 133 patients revealed that none acquired novel RUNX1 mutations during clinical courses. Our findings provide evidence that RUNX1 mutations are associated with distinct biologic and clinical characteristics and poor prognosis in patients with de novo AML. PMID- 19808698 TI - FLT3-ITD up-regulates MCL-1 to promote survival of stem cells in acute myeloid leukemia via FLT3-ITD-specific STAT5 activation. AB - Myeloid cell leukemia-1 (MCL-1) is an essential survival factor for hematopoiesis. In humans, hematopoietic stem cells (HSCs) express MCL-1 at the highest level in response to FMS-like tyrosine kinase-3 (FLT3) signaling. We here show that this FLT3-dependent stem cell maintenance system also plays a critical role in survival of leukemic stem cells (LSCs) in acute myeloid leukemia (AML). The CD34(+)CD38(-) LSC fraction expresses high levels of FLT3 as well as MCL-1, even compared with normal HSCs. Treatment with FLT3 ligand induced further MCL-1 up-regulation in LSCs in all AML cases tested. Interestingly, the group of samples expressing the highest levels of MCL-1 constituted AML with FLT3-internal tandem duplications (ITD). In FLT3-ITD AML cell lines, cells expressed a high level of MCL-1, and an inhibition of MCL-1 induced their apoptotic cell death. A tyrosine kinase inhibitor suppressed MCL-1 expression, and induced apoptosis that was reversed by the enforced MCL-1 expression. Finally, transduction of FLT3-ITD into HSCs strongly activated MCL-1 expression through its signal transducer and activator of transcription 5 (STAT5)-docking domains. This effect was completely abrogated when STAT5 activation was blocked. Thus, the acquisition of FLT3-ITD ensures LSC survival by up-regulating MCL-1 via constitutive STAT5 activation that is independent of wild-type FLT3 signaling. PMID- 19808699 TI - Erythropoietic response to endogenous erythropoietin in premature very low birth weight infants. AB - Despite the common occurrence of anemia in very low birth weight (VLBW) infants, the erythropoiesis and Hb production rates and their relationship to plasma erythropoietin (EPO) concentrations remain unknown in these subjects. To determine these quantities, all blood removed by phlebotomy and administered by red blood cell (RBC) transfusion over the first 30 days of life was recorded in 14 ventilated VLBW infants born at 24 to 28 weeks of gestation. Discarded blood from frequent clinically ordered laboratory blood samples was used to construct plasma EPO, Hb, and RBC concentration-time profiles for each infant. A pharmacodynamic Hb mass balance model that accounted for the dynamic hematological conditions experienced by these infants was simultaneously fitted to the plasma EPO, Hb, and RBC concentrations from each individual subject, while accounting for subject growth. Based on the model estimates, an average of 4.69 g of Hb was produced over the first 30 days of life, compared with 5.97 g removed by phlebotomies and 12.3 g administered by transfusions. These high transfusion amounts were consistent with a relatively short RBC life span and rapidly expanding blood volume with infant growth. The estimated mean body weight-scaled Hb production rate dropped nearly 3-fold after birth to 0.144 g/day x (kg)(3/4). Although only estimated in a subset of the subjects, the mean plasma EPO EC(50) of 28.5 mU/ml and maximal Hb production rate (E(max)) indicated that a severalfold increase in Hb production rate could be achieved with only a modest increase in plasma EPO concentrations. PMID- 19808700 TI - Critical cysteine residues of Kelch-like ECH-associated protein 1 in arsenic sensing and suppression of nuclear factor erythroid 2-related factor 2. AB - Arsenic activates nuclear factor erythroid 2-related factor 2 (Nrf2) to induce phase II and antioxidative genes. Here we analyzed arsenic-Kelch-like ECH associated protein 1 (Keap1) cysteine thiol interaction in Nrf2 activation. Arsenic-based Nrf2 activators, fluorescent biarsenical labeling reagent (FlAsH) and phenylarsine oxide (PAO), were used to probe binding of arsenic to Keap1. Strong fluorescence was observed on binding of FlAsH to purified Keap1. Pretreatment with arsenic, tert-butylhydroquinone (tBHQ), or 2,3 dimercaptopropanol significantly reduced the fluorescent signal. PAO affinity beads effectively pulled down Keap1 in vitro and from hepa1c1c7 cells. Arsenic, tBHQ, free PAO, or cadmium blocked Keap1 pulldown. Furthermore, arsenic and free PAO significantly reduced the free thiol contents of purified or endogenous Keap1. Thus, arsenic, FlAsH, and PAO, as well as tBHQ and cadmium, bind to Keap1 cysteine thiols in a similar fashion. All the domains of Keap1 bound PAO, and the linker region exhibited the highest binding activity. The function of arsenic Keap1 interaction was evaluated in a reconstituted system that mimics endogenous Nrf2 regulation. Mutation of Cys273 or Cys288 in the linker region resulted in high level basal expression of Nrf2 protein. Mutation of Cys151 abolished Nrf2 activation by arsenic. Overexpression of C273A, C288A, or C151A altered the basal and arsenic-induced expression of Nrf2 target genes. The study shows an important role of Cys273 and Cys288 in the suppression of Nrf2 by Keap1 and a critical function of Cys151 in arsenic responsiveness. Our findings support a model in which arsenic binds to different sets of Keap1 cysteine residues to regulate divergent functions in Nrf2 signal transduction. PMID- 19808701 TI - 'There's probably no God; now stop worrying and enjoy yourself'. PMID- 19808702 TI - Protein kinase C delta mediates arterial injury responses through regulation of vascular smooth muscle cell apoptosis. AB - AIMS: A balance between apoptosis and proliferation of vascular smooth muscle cells (VSMC) influences the development of intimal hyperplasia. We have previously demonstrated that protein kinase C delta (PKCdelta) regulates both apoptosis and proliferation of VSMC in vitro. Here we investigate the role of PKCdelta in intimal hyperplasia through gene deletion or overexpression in rodent models of arterial injury. METHODS AND RESULTS: Arterial injury was induced in mice and rats by means of carotid ligation or balloon angioplasty, respectively. Overexpression of PKCdelta was achieved by adenovirus-mediated gene transfer immediately after balloon injury in rat carotid arteries. Levels of PKCdelta protein were profoundly increased in the carotid wall 3-7 days after balloon injury, co-localizing to TUNEL-positive medial cells. When subjected to arterial injury, PKCdelta gene-deficient mice responded with an enhanced intimal hyperplasia accompanied by an 80% reduction in the number of TUNEL-positive cells detected in the injured arteries as compared with their wild-type littermates. Conversely, arterial gene transfer of PKCdelta further increased the arterial expression of PKCdelta, which was associated with a marked increase in apoptosis and reduction of intimal hyperplasia. Neither manipulation led to significant alteration in cell proliferation, suggesting that the function of PKCdelta after arterial injury is predominantly pro-apoptotic. This notion is further supported by our observation of high PKCdelta expression in human restenotic lesions that also co-localized with apoptosis. CONCLUSION: The expression of PKCdelta is upregulated in the arterial wall in response to injury. This induction appears to be a mechanism of arterial response that negatively influences the degree of intimal hyperplasia by stimulating VSMC apoptosis. PMID- 19808703 TI - Zoning for health? The year-old ban on new fast-food restaurants in South LA. AB - A regulation banning new fast-food establishments for one year in Los Angeles, California, was passed unanimously by the city council in July 2008. It was motivated by health concerns and excessive obesity rates in South Los Angeles. However, it might not have had the impact that was intended. This paper reviews the empirical evidence for the regulation and whether it is likely to target the primary levers of obesity. We argue that the premises for the ban were questionable. For example, the density of fast-food chain restaurants per capita is actually higher in other parts of Los Angeles than in South LA. Other changes, such as menu calorie labeling, are likely to have a bigger impact on overweight and obesity. PMID- 19808704 TI - New York City's fight over calorie labeling. AB - In 2006, New York City's Health Department amended the city Health Code to require the posting of calorie counts by chain restaurants on menus, menu boards, and item tags. This was one element of the city's response to rising obesity rates. Drafting the rule involved many decisions that affected its impact and its legal viability. The restaurant industry argued against the rule and twice sued to prevent its implementation. An initial version of the rule was found to be preempted by federal law, but a revised version was implemented in January 2008. The experience shows that state and local health departments can use their existing authority over restaurants to combat obesity and, indirectly, chronic diseases. PMID- 19808705 TI - Calorie labeling and food choices: a first look at the effects on low-income people in New York City. AB - We examined the influence of menu calorie labels on fast food choices in the wake of New York City's labeling mandate. Receipts and survey responses were collected from 1,156 adults at fast-food restaurants in low-income, minority New York communities. These were compared to a sample in Newark, New Jersey, a city that had not introduced menu labeling. We found that 27.7 percent who saw calorie labeling in New York said the information influenced their choices. However, we did not detect a change in calories purchased after the introduction of calorie labeling. We encourage more research on menu labeling and greater attention to evaluating and implementing other obesity-related policies. PMID- 19808706 TI - Functional analysis of putative genes encoding the PIP2 water channel subfamily in Populus trichocarpa. AB - We located fully sequenced putative genes of the plasma membrane intrinsic proteins (PIPs) family in the Populus trichocarpa (Torr. Gray), genome. Of 23 gene candidates, we assigned eight genes to the PIP2 subfamily. All eight putative genes were expressed in vegetative tissues (roots, leaves, bark and wood), and all of them showed water channel activity after being expressed in Xenopus oocytes. Six of eight proteins were affected by mercury ions. No proteins were affected by the presence of nickel or tungsten ions, or by lowering the pH of bathing external solution from 7.4 to 6.5. The presence of copper ions caused seven of eight PIP2 proteins to increase their water transport capacity by as much as 50%. This systematic study of the PIP2 subfamily of proteins in P. trichocarpa provides a basic overview of their activity as water channels and will be a useful reference for future physiological studies of plant water relations that use P. trichocarpa as a model system. PMID- 19808707 TI - Cloning and expression analysis of 14 lipid transfer protein genes from Tamarix hispida responding to different abiotic stresses. AB - Plant lipid transfer proteins (LTPs) are ubiquitous lipid-binding proteins that are involved in various stress responses. In this study, we cloned 14 unique LTP genes (ThLTP 1-14) from Tamarix hispida Willd. (Tamaricaceae) to investigate their roles under various abiotic stress conditions. The expression profiles of the 14 ThLTPs in response to NaCl, polyethylene glycol (PEG), NaHCO(3), CdCl(2) and abscisic acid (ABA) exposure in root, stem and leaf tissues were investigated using real-time RT-PCR. The results showed that all 14 ThLTPs were expressed in root, stem and leaf tissues under normal growth conditions. However, under normal growth conditions, ThLTP abundance varied in each organ, with expression differences of 9000-fold in leaves, 540-fold in stems and 3700-fold in roots. These results indicated that activity and/or physiological importance of these ThLTPs are quite different. Differential expression of the 14 ThLTPs was observed (> 2-fold) for NaCl, PEG, NaHCO(3) and CdCl(2) in at least one tissue indicating that they were all involved in abiotic stress responses. All ThLTP genes were highly induced (> 2-fold) under ABA treatment in roots, stems and/or leaves, and particularly in roots, suggesting that ABA-dependent signaling pathways regulated ThLTPs. We hypothesize that ThLTP expression constitutes an adaptive response to abiotic stresses in T. hispida and plays an important role in abiotic stress tolerance. PMID- 19808708 TI - Excellent functional result in children after correction of anomalous origin of left coronary artery from the pulmonary artery--a population-based complete follow-up study. AB - Surgical strategy to construct a two-coronary system for a patient with anomalous origin of left coronary artery from pulmonary artery (ALCAPA) has evolved with time. Limited long-term follow-up data are available on these children. We report population-based follow-up in children operated on for ALCAPA. In total, 29 patients underwent aortic reimplantation of ALCAPA between 1979 and 2006. Twenty (69%) children were repaired with direct aortic implantation, five (17%) with a modified tubular extension technique, and four (14%) patients with an intrapulmonary baffling technique. Early postoperative mortality (<30 days) was 17%. No late mortality (>30 days) was detected. The median length of follow-up was 11 years (range 10 months-27 years). Global left ventricular function by echocardiography (M-mode) was within normal limits (>30%) in all patients one year after operation. Functionally, 80% of patients were classified in NYHA class I, 20% in NYHA II, and 0% in NYHA classes III/IV at the time of the last examination. Excellent results with good long-term outcome can be achieved in infants with ALCAPA using reimplantation techniques. Normalization of cardiac function is expected within the first year in all operative survivors with a patent coronary system. PMID- 19808709 TI - Warm, beating heart aortic valve replacement in a sickle cell patient. AB - Patients with sickle cell abnormalities undergoing surgery are generally considered to be at greater risk for perioperative complications. We present a 25 year-old woman with sickle cell disease (SCD) and severe aortic insufficiency. A minimally invasive, warm, beating heart approach was adopted to try and minimize the risk of sickling due to cardiopulmonary bypass (CPB), low-flow states, cold cardioplegia and aortic cross-clamping. Compared to classical methods, we believe our technique further reduces the risk of systemic and organ hypothermia and thus, sickling. PMID- 19808710 TI - Marital partner and mortality: the effects of the social positions of both spouses. AB - BACKGROUND: Dimensions of the individual socioeconomic position-education, social class, social status and income-are associated with mortality. Inequalities in death also related to the social position of the household. It is, however, less clear how the socioeconomic position of one marital/cohabiting partner influences the mortality of the other partner. We examine the independent effect on mortality of own and partner's positions regarding these four socioeconomic factors. METHODS: Register data on education, social class, social status and income of both marital/cohabiting partners were collected from the 1990 Census of the employed Swedish population aged 30-59 (N = 1,502,148). Data on all-cause mortality and deaths from cancer and circulatory disease for the subsequent period 1991-2003 were collected from the Cause of Death Register. Relative mortality risks for different socioeconomic groups were estimated by Cox regression. RESULTS: All-cause mortality of both men and women differs by women's education and status and by men's social class and income. For men, the wife's education is more important for the mortality risk than his own education when the man's social class is included in the model. For women, the husband's social class yields larger mortality differences than own occupational measures. Women's education and men's social class and income are particularly important for women's deaths from circulatory diseases. CONCLUSION: The partner's social position has a clear independent association with individual mortality, and women's education and men's social class seem to be particularly important. Suggested explanations of health inequality are not always compatible with the observed relationship between partners' social and economic resources and mortality. PMID- 19808711 TI - A cross-sectional study comparing variation in body surface area and chemotherapy dosing in pediatric oncology using two different methods. AB - PURPOSE: Standardizing body surface area (BSA) determination is essential for avoiding variation in chemotherapy dosage calculations. In this study, we compared variation in BSA calculation using weight and height by the Mosteller formula with weight alone using recently adapted table at a local oncology center. METHODS: Cross-sectional study of pediatric oncology patients presenting to a pediatric oncology clinic over a week period of time. RESULTS: One-hundred consecutive pediatric oncology patients presented to the clinic. The mean BSA calculated by the Mosteller formula was 0.83 m(2) (SD 0.24) and the mean BSA determined by the table (based on weight alone) was 0.82 m(2) (SD 0.25). The mean variation in dosing between the two methods was 1.64% (SD 3.4). Only 13 out of 100 patients (13%) had equal dosing using both methods and 21 out of 100 patients (21%) had dosing variation greater than 5%. When comparing both methods, using paired t-test, the difference was statistically significant (t((99)) = 3.99 and p < 0.001). CONCLUSION: Significant differences in BSAbased chemotherapy dosing exist in our center. The Mosteller method should remain the standard until prospective studies are performed to determine the significance of this dosing variability on toxicity and survival outcome. PMID- 19808712 TI - Vascular metallomics: copper in the vasculature. AB - Owing to recent progress in analytical techniques, metallomics are evolving from detecting distinct trace metals in a defined state to monitor the dynamic changes in the abundance and location of trace metals in vitro and in vivo. Vascular metallomics is an emerging field that studies the role of trace metals in vasculature. This review will introduce common metallomics techniques including atomic absorption spectrometry, inductively coupled plasma-atomic emission spectrometry, inductively coupled plasma-mass spectrometry and X-ray fluorescence spectrometry with a summary table to compare these techniques. Moreover, we will summarize recent research findings that have applied these techniques to human population studies in cardiovascular diseases, with a particular emphasis on the role of copper in these diseases. In order to address the issue of interdisciplinary studies between metallomics and vascular biology, we will review the progress of efforts to understand the role of copper in neovascularization. This recent advance in the metallomics field may be a powerful tool to elucidate the signaling pathways and specific biological functions of these trace metals. Finally, we summarize the evidence to support the notion that copper is a dynamic signaling molecule. As a future direction, vascular metallomics studies may lead to the identification of targets for diagnosis and therapy in cardiovascular disease. PMID- 19808713 TI - An unequal social distribution of peripheral arterial disease and the possible explanations: results from a population-based study. AB - A low socioeconomic status (SES) is associated with higher cardiovascular mortality and morbidity. It has remained unclear whether such a social gradient is evident in peripheral artery disease (PAD) because both diseases show different clinical courses. We looked at the association between education and income with PAD within the population of the Heinz Nixdorf Recall Study (HNRS) including 4738 individuals. In both men and women, the ankle-brachial index (ABI) decreased and the prevalence of PAD (ABI < 0.9) increased with decreasing education and income. Looking only at participants without cardiovascular disease, doctor-diagnosed PAD and media calcinosis participants with low (odds ratio 2.58, 95% confidence interval 1.53-4.34) and median education (1.90, 1.27 2.85) had higher odds for suffering from PAD compared to participants with high education. Additional adjustment for classical arteriosclerosis risk factors reduced the strength of the association while the odds ratios remained elevated. Current smoking, diabetes and BMI seem to be the most relevant mediators. Income does not significantly correlate with events when adjustments for all other risk factors are made. In conclusion, our study demonstrates that PAD is more pronounced in men and women with lower socioeconomic status. Diabetes prevention and smoking prevention and cessation programs need to specifically target individuals of lower income and education. According to our findings, prevention of PAD would benefit extraordinarily. PMID- 19808714 TI - Lower-leg symptoms in peripheral arterial disease are associated with anxiety, depression, and anhedonia. AB - Patients with peripheral arterial disease (PAD) report diverse clinical manifestations that are not always consistent with classic intermittent claudication. We examined the degree to which atypical exertional leg symptoms, intermittent claudication, and exertional leg symptoms that begin at rest were associated with mood states such as anxiety, depressive symptoms, and anhedonia (i.e. lack of positive affect). A cohort of consecutive PAD patients (n = 628) from the Erasmus Medical Center, Rotterdam, The Netherlands, completed the Hospital Anxiety and Depression Scale and the San Diego Claudication questionnaire. The ankle-brachial index and clinical factors were assessed in all patients at baseline. Anxiety was present in 29%, depressive symptoms in 30%, and anhedonia in 28% of patients. Pain at rest was independently associated with anxiety, depressive symptoms, and anhedonia (ORs between 2.5 and 4.0, p /= 30, respectively; p = 0.03. Baseline flow measures were associated with the apnea-hypopnea index but this association was non-significant after adjusting for body mass index. No significant association was observed between measures of SDB and percent flow mediated dilation or hyperemic flow in any model. In conclusion, this study supports a moderate association of SDB and larger baseline brachial artery diameter, which may reflect SDB-induced vascular remodeling. This study does not support a link between SDB and endothelial dysfunction as measured by brachial artery flow-mediated dilation. PMID- 19808719 TI - Risk factors and underlying mechanisms for venous stasis syndrome: a population based case-control study. AB - Venous stasis syndrome may complicate deep vein thrombosis (DVT; i.e. post phlebitic syndrome), but, in most cases, venous stasis syndrome is not post phlebitic. The objective of this study was to determine the risk factors (including prior DVT) for venous stasis syndrome, and to assess venous outflow obstruction and venous valvular incompetence as possible mechanisms for venous stasis syndrome. This was a case-control study nested within a population-based inception cohort. The study population consisted of 232 Olmsted County, MN residents with a first lifetime venous thromboembolism (VTE) and 133 residents without VTE. Measurements included a questionnaire and physical examination for venous stasis syndrome; strain gauge outflow plethysmography, venous continuous wave Doppler ultrasonography and passive venous drainage and refill testing for venous outflow obstruction and venous valvular incompetence. Altogether, 161 (44%), 43 (12%), and 136 (38%) subjects respectively, had venous stasis syndrome, venous outflow obstruction and venous valvular incompetence. Independent risk factors for venous stasis syndrome included increasing patient age and body mass index (BMI), prior DVT, longer time interval since DVT, and varicose veins. Both venous outflow obstruction (p = 0.003) and venous valvular incompetence (p < 0.0001) were strongly associated with venous stasis syndrome. Increasing age and prior DVT were significantly associated with venous outflow obstruction, while prior DVT, varicose veins and venous stasis syndrome diagnosed prior to the incident DVT were significantly associated with venous valvular incompetence. The risks of venous outflow obstruction, venous valvular incompetence and venous stasis syndrome were higher with left leg DVT. In conclusion, increasing patient age and BMI, prior DVT (particularly left leg DVT), longer time interval since DVT and varicose veins are independent risk factors for venous stasis syndrome. Venous stasis syndrome related to DVT is due to venous outflow obstruction and venous valvular incompetence, while venous stasis syndrome related to older age and to varicose veins is due to venous outflow obstruction and to venous valvular incompetence, respectively. PMID- 19808721 TI - Sirolimus-eluting stent placement for refractory renal artery in-stent restenosis: sustained patency and clinical benefit at 24 months. AB - Renal artery stenosis may cause or exacerbate hypertension and renal failure. Percutaneous transluminal renal artery stent placement, increasingly the first line therapy for ostial atherosclerotic renal artery stenosis, can be complicated by in-stent restenosis weeks to months after the procedure. There is currently no consensus for the treatment of in-stent restenosis. Sirolimus-eluting stents have been shown to be effective to treat in-stent restenosis in the coronary circulation. We report a case of sustained 24-month patency after repair of recurrent renal artery in-stent restenosis with use of a sirolimus-eluting stent. PMID- 19808722 TI - Percutaneous revascularization of occluded renal arteries in the setting of acute renal failure. AB - We report the case of a 60-year-old patient with acute renal failure and occluded bilateral renal arteries presenting with acute pulmonary edema and non-ST segment elevation myocardial infarction. The patient required renal replacement therapy with hemodialysis and was subsequently successfully treated with bilateral renal artery stent placement. Marked improvement in renal function was noted within 1 week with freedom from the need for renal replacement therapy at 4 months of follow-up. PMID- 19808723 TI - Ischemic steal syndrome following arm arteriovenous fistula for hemodialysis. AB - Arteriovenous fistulae in the arm are commonly used for hemodialysis in end-stage renal disease. Although physiological steal with reverse flow in the artery distal to the fistula is common, hand ischemia or infarction are rare. The ischemic steal syndrome (hand or forearm ischemia) is usually a result of arterial disease proximal or distal to the fistula and/or poor collateral supply to the hand. The diagnosis is primarily clinical; however, markedly reduced digital pressures and pulse volume recordings support the diagnosis. Management requires imaging for focal stenoses or disease in arteries proximal and distal to the fistula from the aorta to the hand. We present a case caused by subclavian artery occlusion that was initially missed due to focusing investigation only on the fistula. We describe the percutaneous treatments and surgical revisions that attempt to restore flow to the hand without compromising the fistula. PMID- 19808724 TI - An unusual etiology of a subclavian artery aneurysm. AB - A 22-year-old woman presented with a 3-month history of a pulsatile swelling in the right supraclavicular region. A CT angiogram revealed an aneurysm arising from the distal right subclavian artery. At surgery, the subclavian artery was almost entirely replaced by a well-vascularized tumor mass. The vascular tumor along with the native vessel was excised and replaced with a vascular prosthesis. Biopsy was suggestive of an epithelioid hemangioma (EH). In conclusion, an EH is a rare vascular tumor. The presence of vascular hyperplasia with plump endothelial cells protruding into the lumen is the most important discriminator in establishing the diagnosis of EH. Vascular neoplasms presenting as aneurysms should be considered, especially if other etiologies can be excluded. PMID- 19808727 TI - Disparity in outcomes of surgical revascularization for limb salvage. Race and gender are synergistic determinants of vein graft failure and limb loss. Nguyen LL, Hevelone N, Rogers SO, Bandyk DF, Clowes AW, Moneta GL, Lipsitz S, Conte MS. Circulation. 2009; 119: 123-130. AB - STUDY OBJECTIVE: The primary objective of this study was to examine in a post hoc secondary analysis, the interaction of race and gender as they affect graft patency, limb salvage and mortality among patients enrolled in the Project of Ex Vivo Vein Graft Engineering via Transfection III (PREVENT III). PREVENT III was a randomized controlled trial, designed to test the efficacy of edifoligide versus placebo in patients undergoing vein bypass for critical limb ischemia (CLI). POPULATION: PREVENT III enrolled 1404 patients, > 18 years of age with CLI (defined as arterial insufficiency with gangrene, a non-healing ischemic ulcer or rest pain) undergoing vein bypass graft surgery, across 83 North American centres. DESIGN AND METHODS: The primary outcome measure for PREVENT III was time to occurrence of non-technical graft failure resulting in either graft revision or major amputation at 12 months post enrolment. Graft failure was assessed with angiographic or ultrasound surveillance and clinical follow-up at multiple intervals for up to 1 year. For this post hoc study, data from the PREVENT III trial were analysed as an observational cohort for the effect of race and gender on vascular endpoints and patient outcomes. For this analysis, race was dichotomized ('black' versus 'non-black') and race and gender were analysed in individual (dichotomized) and combined groups (black men, black women, non-black men, non-black women). Graft and limb-related endpoints consisted of primary patency (graft patency without intervention), primary assisted patency (graft patency after preventive intervention of a stenosis), secondary patency (graft patency after intervention on a thrombosed graft), major amputation (transtibial or higher) and composite outcomes including amputation-free survival and amputation/revision-free survival. Univariate analysis (using ANOVA and Fisher's exact test) examined the association of race/gender groups with patient demographic characteristics and co-morbidities. Univariate logistic regression models examined the relationship of patient characteristics and 30-day peri operative variables to clinical endpoints. Cox proportional-hazard models were used for 1-year outcomes for graft patency, limb salvage and patient mortality. Propensity score modelling was used to control for 16 covariates (including demographic variables, technical factors, selected co-morbidities and medications) to allow multivariable analysis of the associations and interactions between race and gender with outcomes. Propensity score adjustments nested covariates into the primary outcomes models to analyse the effects of the four combinations of race and gender. RESULTS: Of the 1404 patients enrolled in PREVENT III, 249 black patients (118 women and 131 men) were included. The overall peri-operative (30-day) mortality was 2.7% and no differences were observed between race and gender groups. Although black race and gender were not individually associated with primary patency at 30 days, the subgroup of black men were at increased risk of graft failure at 30 days compared to non-black men (hazard ratio [HR] = 2.96, 95% CI: 1.72-6.06, p < 0.01) and this difference persisted even after adjustments for high-risk grafts (odds ratio [OR] = 3.03, 95% CI: 1.29-7.12, p = 0.01). At 1 year, no significant differences were observed in mortality or primary patency rates among race and gender groups; however, black patients experienced reduced secondary patency compared to non-black patients (HR = 1.49, 95% CI: 1.08-2.06, p = 0.02) and limb salvage (HR = 2.02, 95% CI: 1.27-3.20, p < 0.01). Propensity score models indicated that this disparity was more pronounced among black women (secondary patency HR = 2.02, 95% CI: 1.27-3.20, p < 0.01; major amputation HR = 2.38, 95% CI: 1.18-4.83, p < 0.02). No significant differences were seen between race and gender subgroups in patient survival, amputation-free survival or amputation/revision-free survival. CONCLUSIONS: No differences were observed in mortality or primary patency at 30 days or at 1 year among individual and combined race/gender groups. Black patients, especially black women, were found to have worse secondary patency and limb salvage outcomes after vein bypass surgery for CLI. PMID- 19808725 TI - Novel markers of peripheral arterial disease. AB - Peripheral arterial disease (PAD), a relatively common manifestation of atherosclerotic vascular disease, is associated with significant morbidity and mortality. Although conventional risk factors contribute to the onset and progression of PAD, the role of 'novel' biomarkers in pathways of inflammation, thrombosis, lipoprotein metabolism, and oxidative stress in determining susceptibility to PAD is being increasingly recognized. Validation of novel risk factors for PAD may allow earlier detection, an improved understanding of disease etiology and progression, and the development of new therapies. In this review, we discuss available evidence for associations between novel circulating markers and several aspects of PAD including disease susceptibility, progression, functional limitation, and adverse outcomes. PMID- 19808728 TI - Images in vascular medicine. Herpes vasculitis in systemic lupus erythematosus. PMID- 19808729 TI - Images in vascular medicine. Endograft limb collapse. PMID- 19808730 TI - Global warming factor of municipal solid waste management in Europe. AB - The global warming factor (GWF; CO(2)-eq. tonne(-1) waste) performance of municipal waste management has been investigated for six representative European Member States: Denmark, France, Germany, Greece, Poland and the United Kingdom. The study integrated European waste statistical data for 2007 in a life-cycle assessment modelling perspective. It is shown that significant GWF benefit was achieved due to the high level of energy and material recovery substituting fossil energy and raw materials production, especially in Denmark and Germany. The study showed that, despite strong regulation of waste management at European level, there are major differences in GWF performance among the member states, due to the relative differences of waste composition, type of waste management technologies available nationally, and the average performance of these technologies. It has been demonstrated through a number of sensitivity analyses that, within the national framework, key waste management technology parameters can influence drastically the national GWF performance of waste management. PMID- 19808731 TI - Greenhouse gas accounting and waste management. AB - Accounting of emissions of greenhouse gas (GHG) is a major focus within waste management. This paper analyses and compares the four main types of GHG accounting in waste management including their special features and approaches: the national accounting, with reference to the Intergovernmental Panel on Climate Change (IPCC), the corporate level, as part of the annual reporting on environmental issues and social responsibility, life-cycle assessment (LCA), as an environmental basis for assessing waste management systems and technologies, and finally, the carbon trading methodology, and more specifically, the clean development mechanism (CDM) methodology, introduced to support cost-effective reduction in GHG emissions. These types of GHG accounting, in principle, have a common starting point in technical data on GHG emissions from specific waste technologies and plants, but the limited availability of data and, moreover, the different scopes of the accounting lead to many ways of quantifying emissions and producing the accounts. The importance of transparency in GHG accounting is emphasised regarding waste type, waste composition, time period considered, GHGs included, global warming potential (GWP) assigned to the GHGs, counting of biogenic carbon dioxide, choice of system boundaries, interactions with the energy system, and generic emissions factors. In order to enhance transparency and consistency, a format called the upstream-operating-downstream framework (UOD) is proposed for reporting basic technology-related data regarding GHG issues including a clear distinction between direct emissions from waste management technologies, indirect upstream (use of energy and materials) and indirect downstream (production of energy, delivery of secondary materials) activities. PMID- 19808732 TI - Landfilling of waste: accounting of greenhouse gases and global warming contributions. AB - Accounting of greenhouse gas (GHG) emissions from waste landfilling is summarized with the focus on processes and technical data for a number of different landfilling technologies: open dump (which was included as the worst-case scenario), conventional landfills with flares and with energy recovery, and landfills receiving low-organic-carbon waste. The results showed that direct emissions of GHG from the landfill systems (primarily dispersive release of methane) are the major contributions to the GHG accounting, up to about 1000 kg CO(2)-eq. tonne( -1) for the open dump, 300 kg CO(2)-eq. tonne( -1) for conventional landfilling of mixed waste and 70 kg CO(2)-eq. tonne(-1) for low organic-carbon waste landfills. The load caused by indirect, upstream emissions from provision of energy and materials to the landfill was low, here estimated to be up to 16 kg CO(2)-eq. tonne(-1). On the other hand, utilization of landfill gas for electricity generation contributed to major savings, in most cases, corresponding to about half of the load caused by direct GHG emission from the landfill. However, this saving can vary significantly depending on what the generated electricity substitutes for. Significant amounts of biogenic carbon may still be stored within the landfill body after 100 years, which here is counted as a saved GHG emission. With respect to landfilling of mixed waste with energy recovery, the net, average GHG accounting ranged from about -70 to 30 kg CO(2) eq. tonne(- 1), obtained by summing the direct and indirect (upstream and downstream) emissions and accounting for stored biogenic carbon as a saving. However, if binding of biogenic carbon was not accounted for, the overall GHG load would be in the range of 60 to 300 kg CO(2)-eq. tonne( -1). This paper clearly shows that electricity generation as well as accounting of stored biogenic carbon are crucial to the accounting of GHG of waste landfilling. PMID- 19808733 TI - Integrated waste management as a climate change stabilization wedge. AB - Anthropogenic sources of greenhouse gas emissions are known to contribute to global increases in greenhouse gas concentrations and are widely believed to contribute to climate change. A reference carbon dioxide concentration of 383 ppm for 2007 is projected to increase to a nominal 500 ppm in less than 50 years according to business as usual models. This concentration change is equivalent to an increase of 7 billion tonnes of carbon per year (7 Gt C year(-1)). The concept of a stabilization wedge was introduced by Pacala and Socolow (Science, 305, 968 972, 2004) to break the 7 Gt C year(- 1) into more manageable 1 Gt C year(- 1) reductions that would be achievable with current technology. A total of fifteen possible 'wedges' were identified; however, an integrated municipal solid waste (MSW) management system based on the European Union's waste management hierarchy was not evaluated as a wedge. This analysis demonstrates that if the tonnage of MSW is allocated to recycling, waste to energy and landfilling in descending order in lieu of existing 'business-as-usual' practices with each option using modern technology and best practices, the system would reduce greenhouse gas emissions by more than 1 Gt C year( -1). This integrated waste management system reduces CO(2) by displacing fossil electrical generation and avoiding manufacturing energy consumption and methane emissions from landfills. PMID- 19808734 TI - Collection, transfer and transport of waste: accounting of greenhouse gases and global warming contribution. AB - The collection, transfer and transport of waste are basic activities of waste management systems all over the world. These activities all use energy and fuels, primarily of fossil origin. Electricity and fuel consumptions of the individual processes were reviewed and greenhouse gases (GHG) emissions were quantified. The emission factors were assigned a global warming potential (GWP) and aggregated into global warming factors (GWFs), which express the potential contribution to global warming from collection, transport and transfer of 1 tonne of wet waste. Six examples involving collection, transfer and transport of waste were assessed in terms of GHG emissions, including both provision and use of energy. (GHG emissions related to production, maintenance and disposal of vehicles, equipment, infrastructure and buildings were excluded.) The estimated GWFs varied from 9.4 to 368 kg CO(2)-equivalent (kg CO(2)-eq.) per tonne of waste, depending on method of collection, capacity and choice of transport equipment, and travel distances. The GHG emissions can be reduced primarily by avoiding transport of waste in private cars and by optimization of long distance transport, for example, considering transport by rail and waterways. PMID- 19808735 TI - Determination of reliable CO2 emission factors for waste-to-energy plants. AB - At Vienna University of Technology, the so-called balance method (BM) was developed to determine fossil and biogenic CO(2) emissions from waste-to-energy (WTE) plants. Meanwhile, the BM has been routinely applied to several WTE plants for some years, providing a large set of data. The average site-specific emission factors for fossil CO(2) were found to be in the range of 260- 780 kg CO(2) t(-1) waste, and 30-67 kg CO(2) GJ(-1) energy of the waste incinerated. These values are significantly different from the values that are found in the literature. Our results show that there is no such typical emission factor for WTE which could be applied to national CO(2) measurements or accurate emission trading. This study reveals that instead of generic emission factors the BM can be used as a standard for WTE plants, since its application requires either no or only a few additional installations. PMID- 19808736 TI - Kinetic models for the adsorption of lead ions by steel slag. AB - Batch experiments were carried out to investigate the kinetics of adsorption of lead ions by steel slag on the basis of the external diffusion, intraparticle diffusion and adsorption reaction model (pseudo-first-order, pseudo-second order). The results showed that the controlling step for the adsorption kinetics changed with experimental parameters varied. When the particle size of steel slag was larger than 120 mesh, intraparticle diffusion of Pb(2+) was the controlling step; when the initial concentration of Pb(2+) was less than 150 mg L(-1) or the shaking rate was lower than 150 rpm, external diffusion of Pb(2+) was promoted. Contrary to the former experimental conditions, the adsorption reaction was the controlling step, and the adsorption followed second-order kinetics, with an adsorption rate constant of 13.26 g mg(-1) min(- 1). The adsorption isotherm of Pb(2+) with steel slag followed the Langmuir model, with a correlation coefficient of 0.99. PMID- 19808737 TI - Effect of boron waste on the properties of mortar and concrete. AB - Utilization of by-products or waste materials in concrete production are important subjects for sustainable development and industrial ecology concepts. The usages as mineral admixtures or fine aggregates improve the durability properties of concrete and thus increase the economic and environmental advantages for the concrete industry. The effect of clay waste (CW) containing boron on the mechanical properties of concrete was investigated. CW was added in different proportions as cement additive in concrete. The effect of CW on workability and strength of concrete were analysed by fresh and hardened concrete tests. The results obtained were compared with control concrete properties and Turkish standard values. The results showed that the addition of CW had a small effect upon the workability of the concrete but an important effect on the reduction of its strength. It was observed that strength values were quite near to that of control concrete when not more than 10% CW was used in place of cement. In addition to concrete specimens, replacing cement with CW produced mortar specimens, which were investigated for their strength and durability properties. The tests of SO( 4) (2-) and Cl(-) effect as well as freeze-thaw behaviour related to the durability of mortar were performed. Consequently, it can be said that some improvements were obtained in durability properties even if mechanical properties had decreased with increasing CW content. PMID- 19808738 TI - Chemical precipitation for controlling nitrogen loss during composting. AB - Aimed at controlling the nitrogen loss during composting, the mixture of magnesium hydroxide (Mg(OH)( 2)) and phosphoric acid (H(3)PO(4)) (molar ratio 1:2) were utilized as additives to avoid increasing total salinity. In trial TA, the additives were put into absorption bottles connecting with a gas outlet of fermentor (ex situ method); in trial TB, the additives were directly added to the composting materials (in situ method). During the 26 day composting period, the temperature, pH, total organic carbon (TOC), total nitrogen (TN), ammonium nitrogen (NH(4)(+)-N), total phosphorus (TP), available phosphorus (AP) and germination index (GI) were measured. The experimental results show that the additives reduced the pH, while NH( 4)(+)-N and TN were obviously improved. NH(4)( +)-N was 11.9 g kg(-1) and 3 g kg(- 1) in amended compost trial (TB) and unamended compost trial (TA), respectively; TN increased from 26.5 g kg(-1) to 40.3 g kg(-1) in TB and increased from 26.5 g kg( -1) to 26.8 g kg(-1) in TA. Analysis of the TOC and carbon mass revealed that absorbents accelerated the degradation of organic matter. The germination index test showed the maturity of TB (102%) was better than TA (82%) in final compost. Furthermore, TP and AP were also obviously improved. X-ray diffraction analysis of precipitation showed that the precipitation in absorption bottle of TA was newberyite (MgHPO( 4) 3H(2)O), however, the crystal in the TB compost was struvite (MgNH(4)PO(4) 6H(2)O: magnesium ammonium phosphate). These results indicated that Mg(OH)(2) and H(3)PO( 4) could reduce the ammonia emission by struvite crystallization reaction. Optimal conditions for struvite precipitation should be determined for different systems. PMID- 19808739 TI - Energy use and recovery in waste management and implications for accounting of greenhouse gases and global warming contributions. AB - The energy system plays an essential role in accounting of greenhouse gas (GHG) emissions from waste management systems and waste technologies. This paper focuses on energy use and energy recovery in waste management and outlines how these aspects should be addressed consistently in a GHG perspective. Essential GHG emission data for the most common fuels, electricity and heat are provided. Average data on electricity provision show large variations from country to country due to different fuels being used and different efficiencies for electricity production in the individual countries (0.007-1.13 kg CO(2)-eq. kWh( 1)). Marginal data on electricity provision show even larger variations (0.004-3 kg CO(2)-eq. kWh( -1)). Somewhat less variation in GHG emissions is being found for heat production (0.01-0.69 kg CO(2)-eq. kWh( -1)). The paper further addresses allocation principles and the importance of applying either average or marginal energy data, and it discusses the consequences of introducing reduction targets on CO( 2) emissions. All discussed aspects were found to significantly affect the outcome of GHG accounts suggesting transparent reporting to be critical. Recommendations for use of average/marginal energy data are provided. PMID- 19808740 TI - 'Time is brain' also in multiple sclerosis. PMID- 19808741 TI - 25-hydroxyvitamin D in cerebrospinal fluid during relapse and remission of multiple sclerosis. AB - Hypovitaminosis D may play a role in multiple sclerosis (MS), but little is known about intrathecal vitamin D. 25-Hydroxyvitamin D was measured in cerebrospinal fluid and sera from 36 patients with relapsing-remitting MS, 20 patients with other inflammatory neurological diseases and 18 patients with non-inflammatory neurological diseases with liquid chromatography-mass spectrometry. There were no significant differences in cerebrospinal fluid concentrations of 25 hydroxyvitamin D, but the cerebrospinal fluid:serum ratio was significantly lower in MS compared with other inflammatory neurological diseases (p=0.0012) and non inflammatory neurological diseases (p=0.041) patients. The concentrations of 25 hydroxyvitamin D in cerebrospinal fluid and serum were positively correlated and their ratio was similar to that of albumin. Neither the concentrations of 25 hydroxyvitamin D in cerebrospinal fluid or serum nor their ratio were associated with the presence of relapses or gadolinium-enhanced lesions. These results do not support that 25-hydroxyvitamin D is actively transported to the cerebrospinal fluid, or that the cerebrospinal fluid or serum levels or their ratio exert a major impact on MS activity. PMID- 19808742 TI - Interpretation of internal quality control: time for a re-think? PMID- 19808743 TI - Epidemics of mold poisoning past and present. AB - Molds are ubiquitous throughout the biosphere of planet earth and cause infectious, allergic, and toxic diseases. Toxic diseases arise from exposure to mycotoxins produced by molds. Throughout history, there have been a number of toxic epidemics associated with exposure to mycotoxins. Acute epidemics of ergotism are caused by consumption of grain infested by fungi of the genus Claviceps, which produce the bioactive amine ergotamine that mimics the neurotransmitters norepinephrine, serotonin, and dopamine. Acute aflatoxin outbreaks have occurred from ingestion of corn stored in damp conditions that potentiate growth of the molds of the species Aspergillus. Contemporary construction methods that use cellulose substrates such as fiber board and indoor moisture have caused an outbreak of contaminated buildings with Stachybotrys chartarum, with the extent of health effects still a subject of debate and ongoing research. This article reviews several of the more prominent epidemics and discusses the nature of the toxins. Two diseases that were leading causes of childhood mortality in England in the 1970s and vanished with changing dietary habits, putrid malignant fever, and slow nervous fever were most likely toxic mold epidemics. PMID- 19808744 TI - Fungal exposure endocrinopathy in sinusitis with growth hormone deficiency: Dennis-Robertson syndrome. AB - A retrospective study was carried out on 79 patients with a history of mold exposure, fatigue, and chronic rhinosinusitis (CRS) to determine whether there is a causal relationship between fungal exposure and chronic sinusitis, fatigue, and anterior hypopituitarism, especially growth hormone deficiency (GHD). Of the patients, 94% had a history of CRS, endoscopically and/or computed tomography (CT) confirmed; 100% had chronic fatigue and 100% had either significant history of indoor mold exposure and/or positive mold plate testing as measured by settle plates, with an average colony count of 21 (0-4 normal). A total of 62 had positive mold plate testing and 17 had positive history of mold exposure. Of 75, 73 (97.3%) had positive serum immunoglobulin G (IgG)-specific antibodies to fungal antigens. Out of 8, 7 were positive for urinary trichothecenes. Resting levels of insulin-like growth factor 1 (IGF-1) averaged 123 ng/mL (range 43-285, normal 88-249 ng/mL). Despite normal resting levels of IGF-1, significant deficiency of serum human growth hormone (GH) was confirmed by insulin tolerance test (ITT) in 40 of 50 tested. In all, 51% (40/79) were GH deficient. Primary or secondary hypothyroidism in T3 and/or T4 was seen in 81% (64/79) patients; 75% (59/79) had adrenocorticotrophic hormone (ACTH) deficiency. Fungal exposure endocrinopathy likely represents the major cause of GHD, affecting approximately 4.8 million people compared to approximately known 60,000 cases from all other causes. A literature review indicates a possible mechanism of GHD in fungal exposure is that the fungal glucan receptors in the lenticulostellate cells of the anterior pituitary bind to fungal cells wall glucans and activate the innate immune system, which activates macrophages that destroy the fungus and lenticulostellate tissue. Treatment of patients included normal saline nasal irrigations, antifungal and antibiotic nasal sprays, appropriate use of oral antibiotics and antifungals, facial steamer with CitriDrops. Thymate and/or Intramax vitamin supplements, hormone replacement, and reduction of indoor mold levels. Resolution of rhinosinusitis was seen in 93% (41 of 45) of the patients who achieved a mold count by settling plates of 0-4 colonies. Thirty patients were unable to lower their mold counts below four colonies and had various degrees of mucosal disease and fatigue remaining. Fatigue was improved in all 37 patients who received GH and cortisol and/or thyroid hormone, which were deficient. Fatigue was partially relieved in 7 of the 37 who did not achieve mold counts of fewer than four colonies. PMID- 19808745 TI - Effects of epidural anesthesia on intrathoracic blood volume and extravascular lung water during on-pump cardiac surgery. AB - BACKGROUND: The most important side effect of epidural anesthesia is hypotension with functional hypovolemia. Aggressive infusion therapy can reduce the hypotension effect. However, in conjunction with cardiopulmonary bypass, it can increase acute lung injury. We hypothesized that epidural anesthesia, by reducing cardiac sympathetic tonus, with subsequent better pulmonary flow, does not increase lung interstitial fluids. METHODS: Sixty patients undergoing coronary artery bypass graft (CABG) surgery with cardiopulmonary bypass (CPB) were randomized to combined general anesthesia with epidural anesthesia / analgesia, (EA) group, and to general anesthesia with i/v opiate analgesia, (GA) group. Patients in the EA group received a high thoracic epidural, preoperatively. Intraoperatively, 0.25% bupivacaine 8 mL/h was infused and general anesthesia with sevoflurane was followed by bupivacaine infusion for 48 hours postoperatively. General anesthesia in the GA group was with sevoflurane and fentanyl 10 - 12 microg/kg and analgesia with pethidinum 0.1 - 0.4 mg/kg i.v. postoperatively. Global end-diastolic volume index (GEDI), intrathoracic blood volume index (ITBI) and extravascular lung water index (ELWI) were measured before anesthesia, before CPB and 15, 60, 180, 600 min. and 24 hr after CPB. Duration of mechanical lung ventilation was registered in both groups. RESULTS: ITBI and GEDI were significantly higher in the EA group at all time points of measurement (ITBI 945.6+/-146.4 ml/m(2) and 870.6+/-146.5 ml/m( 2) vs. 1118+/ 153.2 ml/m(2) and 1020+/-174.9 ml/m( 2); GEDI 720+/-96.19 ml/m(2) and 775.0+/ 159.5 ml/m( 2) vs. 805.4+/-97.59 ml/m(2) and 888+/-117.3 ml/m( 2)). GEDI was significantly lower in the GA group compared with baseline (801.9+/-132.4 ml/m(2) vs. 695+/-169.2 mL/m(2)). ELWI was significantly higher in the GA group (7.233+/ 1.35 ml/kg and 7.333+/-1.32 ml/kg vs. 8.533+/-1.45 ml/kg and 8.633+/-1.71 ml/kg), but without significant changes in the EA group. Duration of mechanical lung ventilation was shorter in the EA group (663.7+/-98.39 min. vs. 362.2+/-33.72 min.). CONCLUSIONS: Epidural anesthesia / analgesia does not increase interstitial lung fluids by increasing intrathoracic blood volume or the amount of infusion fluids in patients undergoing cardiac surgery under cardiopulmonary bypass. There is, also, a decreased duration of mechanical lung ventilation. PMID- 19808746 TI - Serum level of sCD163, a soluble receptor for hemoglobin, is influenced by cardiac surgery. AB - BACKGROUND: The scavenger receptor for complexes hemoglobin-haptoglobin (CD163), which is expressed on monocytes/ macrophages, is shed to the body fluids in a soluble form (sCD163). OBJECTIVES: To evaluate the dynamics of sCD163 in the blood of patients undergoing cardiac surgery. PATIENTS AND METHODS: Sixty-one adult patients who underwent coronary artery bypass grafting (CABG) were enrolled in the study. They were assigned to undergo CABG using either cardiopulmonary bypass (CPB), "on-pump", (22 patients), modified CPB, mini "on-pump", (17 patients) or without CPB, "off-pump", (22 patients) surgery. Serum levels of sCD163 in venous blood samples taken before and after surgery, and during an early postoperative period, were evaluated by Macro 163(TM) diagnostic kit (IQ Products, Groningen, NL). RESULTS: Compared to the preoperative levels ("on pump"; 344 ng/mL, "off-pump"; 314.5 ng/mL, mini-invasive "on-pump"; 336.5 ng/mL) serum levels were elevated at the finish of surgery, reaching maximum at the 1(st) postoperative day ("onpump"; 658 ng/mL; p<0.05, "off-pump"; 810.5 ng/mL; p<0.01; mini-invasive "on-pump"; 663 ng/mL; non-significant).No significant differences regarding the serum levels of sCD163 between different surgical approaches were found. CONCLUSION: Serum level of sCD163 scavenger molecule for hemoglobin is elevated at the end of surgery and at the 1(st) postoperative day, being little influenced by cardiopulmonary bypass. PMID- 19808747 TI - Venoarterial extracorporeal membrane oxygenation in adult patients: predictors of mortality. AB - BACKGROUND: Extracorporeal membrane oxygenation is a cardiopulmonary supportive therapy. In this study, we reviewed our experience with extracorporeal membrane oxygenation support and tried to identify measurable values which might predict in-hospital mortality. METHODS: From January 2004 through December 2008, 50 of 21,298 adult patients received venoarterial extracorporeal membrane oxygenation. We retrospectively analyzed clinical records of these 50 consecutive patients. Details of demographics, preoperative measurements, clinical characteristics at the time of extracorporeal membrane oxygenation implantation, extracorporeal membrane oxygenation-related complications and in-hospital mortality were collected. Logistic regression analyses were performed to investigate predictors of mortality. A p-value < or = 0.05 was accepted as significant. RESULTS: Thirty eight patients were weaned from extracorporeal membrane oxygenation and 33 patients survived to discharge. The overall survival rate was 66%. In a multiple logistic regression analysis, blood lactate level before initiation of extracorporeal membrane oxygenation was a risk factor associated with in-hospital mortality (OR 1.27 95% CI 1.042-1.542). To evaluate the utility of the lactate in predicting mortality, a conventional receiver operating characteristic curve was produced. Sensitivity and specificity were optimal at a cut-off point of 12.6 mmol/L, with an area under the curve of 0.752. The positive and negative predictive values were 73.3% and 83.9%, respectively. CONCLUSIONS: Extracorporeal membrane oxygenation is a justifiable alternative treatment for postoperative refractory cardiac and pulmonary dysfunction which could rescue more than sixty percent of otherwise fatal patients. Patients with pre-extracorporeal membrane oxygenation lactate levels above 12.6 mmol/L are at higher risks for in-hospital death. Evidence-based therapy for this group of high risk patients is needed. PMID- 19808748 TI - We must defend personal continuity in primary care. PMID- 19808750 TI - Charity and drug company work together to develop cure for Chagas' disease. PMID- 19808753 TI - Margaret Allen: a force in humanising the American healthcare system. Interview by Zosia Kmietowicz. PMID- 19808754 TI - The tyranny of power: is there a better way to calculate sample size? PMID- 19808755 TI - The safety of propofol. PMID- 19808756 TI - Escherichia coli O157. PMID- 19808759 TI - Medical relief efforts in Sumatra and Pacific islands are stepped up. PMID- 19808762 TI - Nobel medicine prize is won by scientists for work on chromosomal telomeres. PMID- 19808764 TI - Prognosis of low back pain in primary care. PMID- 19808765 TI - Common mental disorder and obesity: insight from four repeat measures over 19 years: prospective Whitehall II cohort study. AB - OBJECTIVES: To examine potential reciprocal associations between common mental disorders and obesity, and to assess whether dose-response relations exist. DESIGN: Prospective cohort study with four measures of common mental disorders and obesity over 19 years (Whitehall II study). SETTING: Civil service departments in London. PARTICIPANTS: 4363 adults (28% female, mean age 44 years at baseline). MAIN OUTCOME: Common mental disorder defined as general health questionnaire "caseness;" overweight and obesity based on Word Health Organization definitions. RESULTS: In models adjusted for age, sex, and body mass index at baseline, odds ratios for obesity at the fourth screening were 1.33 (95% confidence interval 1.00 to 1.77), 1.64 (1.13 to 2.36), and 2.01 (1.21 to 3.34) for participants with common mental disorder at one, two, or three preceding screenings compared with people free from common mental disorder (P for trend<0.001). The corresponding mean differences in body mass index at the most recent screening were 0.20, 0.31, and 0.50 (P for trend<0.001). These associations remained after adjustment for baseline characteristics related to mental health and exclusion of participants who were obese at baseline. In addition, obesity predicted future risk of common mental disorder, again with evidence of a dose-response relation (P for trend=0.02, multivariable model). However, this association was lost when people with common mental disorder at baseline were excluded (P for trend=0.33). CONCLUSIONS: These findings suggest that in British adults the direction of association between common mental disorders and obesity is from common mental disorder to increased future risk of obesity. This association is cumulative such that people with chronic or repeat episodes of common mental disorder are particularly at risk of weight gain. PMID- 19808766 TI - Prognosis for patients with chronic low back pain: inception cohort study. AB - OBJECTIVES: To describe the course of chronic low back pain in an inception cohort and to identify prognostic markers at the onset of chronicity. DESIGN: Inception cohort study with one year follow-up. SETTING: Primary care clinics in Sydney, Australia. PARTICIPANTS: The study sample was a subcohort of an inception cohort of 973 consecutive patients presenting to primary care with acute low back pain (<2 weeks' duration). 406 participants whose pain persisted for three months formed the inception cohort of patients with chronic low back pain. MAIN OUTCOME MEASURES: Outcomes and putative predictors measured at initial presentation, onset of chronicity (study entry), and follow-up at nine and 12 months. Recovery was determined from measures of pain intensity, disability, and work status. The association between potential prognostic factors and time to recovery was modelled with Cox regression. RESULTS: Completeness of follow-up was 97% of total person time for all outcomes. The cumulative probability of being pain-free was 35% at nine months and 42% at 12 months and for complete recovery was 35% at nine months and 41% at 12 months. Of the 259 participants who had not recovered from pain related disability at entry to the chronic study, 47% had recovered by 12 months. Previous sick leave due to low back pain, high disability levels or high pain intensity at onset of chronicity, low levels of education, greater perceived risk of persistent pain, and being born outside Australia were associated with delayed recovery. CONCLUSION: More than one third of patients with recent onset, non-radicular chronic low back pain recover within 12 months. The prognosis is less favourable for those who have taken previous sick leave for low back pain, have high disability levels or high pain intensity at onset of chronic low back pain, have lower education, perceive themselves as having a high risk of persistent pain, and were born outside Australia. PMID- 19808767 TI - Obesity and depression or anxiety. PMID- 19808768 TI - Partial protection of seasonal trivalent inactivated vaccine against novel pandemic influenza A/H1N1 2009: case-control study in Mexico City. AB - OBJECTIVE: To evaluate the association of 2008-9 seasonal trivalent inactivated vaccine with cases of influenza A/H1N1 during the epidemic in Mexico. DESIGN: Frequency matched case-control study. SETTING: Specialty hospital in Mexico City, March to May 2009. PARTICIPANTS: 60 patients with laboratory confirmed influenza A/H1N1 and 180 controls with other diseases (not influenza-like illness or pneumonia) living in Mexico City or the State of Mexico and matched for age and socioeconomic status. MAIN OUTCOME MEASURES: Odds ratio and effectiveness of trivalent inactivated vaccine against influenza A/H1N1. RESULTS: Cases were more likely than controls to be admitted to hospital, undergo invasive mechanical ventilation, and die. Controls were more likely than cases to have chronic conditions that conferred a higher risk of influenza related complications. In the multivariate model, influenza A/H1N1 was independently associated with trivalent inactivated vaccine (odds ratio 0.27, 95% confidence interval 0.11 to 0.66) and underlying conditions (0.15, 0.08 to 0.30). Vaccine effectiveness was 73% (95% confidence interval 34% to 89%). None of the eight vaccinated cases died. CONCLUSIONS: Preliminary evidence suggests some protection from the 2008-9 trivalent inactivated vaccine against pandemic influenza A/H1N1 2009, particularly severe forms of the disease, diagnosed in a specialty hospital during the influenza epidemic in Mexico City. PMID- 19808769 TI - The future of influenza vaccines. PMID- 19808770 TI - Should patients with high-risk soft tissue sarcoma receive adjuvant chemotherapy? AB - Soft tissue sarcoma is a malignant connective tissue tumor that may arise anywhere in the body and from diverse mesenchymal elements. Its incidence is approximately 30 per million persons. The majority of patients with soft tissue sarcoma present with potentially life-threatening disease, and complete resection to obtain specimen margins free of tumor and radiation offer the best chance for local disease control. The risk of relapse and death from disease rises with increasing tumor stage, grade, and size. Adjuvant chemotherapy has been studied as a means to decrease the risk for disease recurrence in patients with localized soft tissue sarcoma at diagnosis, but the majority of trials reported on have been hampered by patient heterogeneity, low patient accrual, and short follow-up. Meta-analysis and reviews of institutional large series, in efforts to overcome some of the limitations, suggest that doxorubicin with ifosfamide reduces the risk for sarcoma recurrence and death in selected patients with high-grade, large, and chemotherapy-sensitive sarcoma subtypes to a clinically meaningful degree. In multiple analyses, patients with high-risk soft tissue sarcoma treated with chemotherapy have a >10% absolute lower risk for disease recurrence and longer disease-specific survival than patients treated without chemotherapy. In the absence of conclusive results from an adequately powered, randomized, controlled clinical trial, the available data support the use of chemotherapy in the management of high-risk, localized, soft tissue sarcoma. PMID- 19808771 TI - Adjuvant chemotherapy in localized soft tissue sarcomas: still not proven. AB - Soft tissue sarcoma is a rare and heterogeneous group of tumors in terms of histological subtypes, molecular alterations, clinical presentation, and prognosis. Yet, these tumors are most often treated similarly in the localized phase. The standard treatment of these patients requires multidisciplinary management, in particular, careful diagnostic procedures and surgery by an expert physician, preceded or followed by external radiotherapy. The utility of adjuvant chemotherapy has been explored in 14 trials comparing adjuvant chemotherapy with no treatment. Several trials reported a lower risk for local relapse and lower risk for metastatic relapse, but only a few small trials reported longer overall survival. A meta-analysis of all trials failed to demonstrate a significant difference in the relapse-free survival (RFS) or overall survival rates. Two additional trials, reported afterward, presented conflicting results, with a significant benefit in terms of the RFS rate for the trial of the Italian Sarcoma Group, but no difference in the RFS or overall survival rate in the most recent European Organization for Research and Treatment of Cancer trial. We conclude that adjuvant chemotherapy has not been proven to improve the outcome of an unselected population of patients. Several hypotheses are proposed to account for this observation. PMID- 19808772 TI - The assessment and management of delirium in cancer patients. AB - Delirium remains the most common and distressing neuropsychiatric complication in patients with advanced cancer. Delirium causes significant distress to patients and their families, and continues to be underdiagnosed and undertreated. The most frequent, consistent, and, at the same time, reversible etiology is drug-induced delirium resulting from opioids and other psychoactive medications. The objective of this narrative review is to outline the causes of delirium in advanced cancer, especially drug-induced delirium, and the diagnosis and management of opioid induced neurotoxicity. The early symptoms and signs of delirium and the use of delirium-specific assessment tools for routine delirium screening and monitoring in clinical practice are summarized. Finally, management options are reviewed, including pharmacological symptomatic management and also the provision of counseling support to both patients and their families to minimize distress. PMID- 19808773 TI - Paternity analysis-based inference of pollen dispersal patterns, male fecundity variation, and influence of flowering tree density and general flowering magnitude in two dipterocarp species. AB - BACKGROUND AND AIMS: Knowledge of pollen dispersal patterns and variation of fecundity is essential to understanding plant evolutionary processes and to formulating strategies to conserve forest genetic resources. Nevertheless, the pollen dispersal pattern of dipterocarp, main canopy tree species in palaeo tropical forest remains unclear, and flowering intensity variation in the field suggests heterogeneity of fecundity. METHODS: Pollen dispersal patterns and male fecundity variation of Shorea leprosula and Shorea parvifolia ssp. parvifolia on Peninsular Malaysian were investigated during two general flowering seasons (2001 and 2002), using a neighbourhood model modified by including terms accounting for variation in male fecundity among individual trees to express heterogeneity in flowering. KEY RESULTS: The pollen dispersal patterns of the two dipterocarp species were affected by differences in conspecific tree flowering density, and reductions in conspecific tree flowering density led to an increased selfing rate. Active pollen dispersal and a larger number of effective paternal parents were observed for both species in the season of greater magnitude of general flowering (2002). CONCLUSIONS: The magnitude of general flowering, male fecundity variation, and distance between pollen donors and mother trees should be taken into account when attempting to predict the effects of management practices on the self-fertilization and genetic structure of key tree species in tropical forest, and also the sustainability of possible management strategies, especially selective logging regimes. PMID- 19808774 TI - Effects of a recombinant FVIIa analogue, NN1731, on blood loss and survival after liver trauma in the pig. AB - BACKGROUND: We considered whether haemorrhage after a liver trauma would be reduced by early administration of a pro-haemostatic agent and evaluated the effect of i.v. vs i.m. administration of the coagulation factor VIIa analogue NN1731 on haemorrhage after a liver trauma in the pig. METHODS: The pharmacokinetics of i.v. and i.m. NN1731 was evaluated in eight minipigs, and the effects of dose and administration route of NN1731 (i.v. 180 microg kg(-1), n=6; i.m. 540 microg kg(-1), n=4, or 2000 microg kg(-1), n=6) vs vehicle (n=16) were studied on a liver laceration injury in pigs. To simulate a pre-hospital setting, the administration of NN1731 was delayed by 1 min for i.m. administration and 7 min for i.v. administration, at which time fluid resuscitation also began. RESULTS: In the minipigs, NN1731 exposure was similar after i.v. 180 microg kg( 1) and i.m. 540 microg kg(-1), with a bioavailability of approximately 35%. The injury and blood loss at 7 min was comparable between the four groups of pigs; however, after 60 min, the blood loss was lower in the i.v. treated animals: 1.3 (0.3) (i.v.) vs 2.2 (0.8) litres (i.m.(540), i.m.(2000), and vehicle) (P<0.001). Also, the survival time was increased: 117 (14) (i.v.) vs 84 (28) min (i.m.(540), i.m.(2000), and vehicle) (P<0.001). CONCLUSIONS: After a liver trauma in the pig, i.v. administration of NN1731 reduced the bleeding and increased the survival time. In contrast, i.m. administration had no effect, presumably because reduced muscle perfusion during haemorrhage reduced the uptake of NN1731. PMID- 19808775 TI - Stress sensitivity is increased in transgenic rats with low brain angiotensinogen. AB - AT(1) blockers attenuate hypothalamo-pituitary-adrenal (HPA) axis reactivity in hypertension independently of their potency to lower blood pressure. A reduced pituitary sensitivity to CRH and a downregulation of hypothalamic CRH expression have been suggested to influence HPA axis activity during chronic AT(1) blockade. This study was aimed at confirming the role of central angiotensin II in regulating HPA reactivity by using the transgenic rat TGR(ASrAOGEN), a model featuring low levels of brain angiotensinogen. Different stress tests were performed to determine HPA reactivity in TGR(ASrAOGEN) and appropriate controls. In TGR(ASrAOGEN), blood pressure was diminished compared to controls. The corticosterone response to a CRH or ACTH challenge and a forced swim test was more distinct in TGR(ASrAOGEN) than it was in controls and occurred independently of a concurrent enhancement in ACTH. Using quantitative real-time PCR, we found increased mRNA levels of melanocortin 2 (Mc2r) and AT(2) receptors (Agtr2) in the adrenals of TGR(ASrAOGEN), whereas mRNA levels of Crh, Pomc, and AT(1) receptors (Agtr1) remained unchanged in hypothalami and pituitary glands. Since stress responses were increased rather than attenuated in TGR(ASrAOGEN), we conclude that the reduced HPA reactivity during AT(1) blockade could not be mimicked in a specific transgenic rat model featuring a centrally inactivated renin-angiotensin aldosterone system. The ACTH independency of the enhanced corticosterone release during CRH test and the enhanced corticosterone response to ACTH rather indicates an adrenal mechanism. The upregulation of adrenal MC2 and AT(2) receptors seems to be involved in the stimulated facilitation of adrenal corticosterone release for effectuating the stimulated stress responses. PMID- 19808776 TI - Pro-arrhythmia in atrial fibrillation suppression pacing algorithms. AB - Atrial overdrive pacing algorithms have been partially effective in controlling atrial fibrillation (AF). A 76-year-old man with history of a cardiomyopathy and paroxysmal AF underwent implant of a dual-chamber ICD. After enabling preferential pacing (PP) algorithms, marked control of his AF was demonstrated, but with inappropriate ICD shocks secondary to a typical AV nodal re-entrant tachycardia. After successful slow pathway modification, no further episodes were documented with suppression of his AF burden with PP algorithms enabled. PMID- 19808777 TI - Effect of RF-amide-related peptide-3 on luteinizing hormone and follicle stimulating hormone synthesis and secretion in ovine pituitary gonadotropes. AB - GnRH provides the primary stimulus for the reproductive axis, but original work also revealed the existence of a gonadotropin-inhibitory hormone (GnIH) in birds. In mammals, GnIH properties are displayed by a hypothalamic dodecapeptide, which is a member of the RF-amide family, namely RF-amide-related peptide (RFRP)-3. This peptide inhibits GnRH-stimulated gonadotropin secretion from ovine pituitary cells in culture, but it is not known whether there are effects on gonadotropin synthesis. The aim of the present study was to determine the effects of RFRP-3 on the expression of genes for beta-subunits of the gonadotropins in ovine pituitary cells from gonadectomized ewes and rams. Cells in primary culture were given GnRH or vehicle pulses every 8 h for 24 h with and without RFRP-3 treatment. GnRH stimulated LH and FSH secretion, which was reduced by RFRP-3. Quantitative real time PCR revealed increased expression of LHbeta and FSHbeta subunit genes after GnRH treatment and a specific reduction in expression after RFRP-3 treatment. There was no effect on the expression of GH, proopiomelanocortin, or prolactin genes. Western blotting showed that GnRH stimulated phosphorylation of ERK (phospho-ERK-1/2), and this effect was abolished by RFRP-3. We conclude that RFRP 3 acts on the pituitary gonadotropes to inhibit synthesis of the gonadotropins, and this effect may be mediated by a reduction in the GnRH-stimulated second messenger phospho-ERK-1/2. PMID- 19808778 TI - Facilitative glucose transporter 9 expression affects glucose sensing in pancreatic beta-cells. AB - Facilitative glucose transporters (GLUTs) including GLUT9, accelerate the facilitative diffusion of glucose across the plasma membrane. Studies in GLUT2 deficient mice suggested the existence of another GLUT in the mammalian beta-cell responsible for glucose sensing. The objective of this study was to determine the expression and function of GLUT9 in murine and human beta-cells. mRNA and protein expression levels were determined for both isoforms of GLUT9 in murine and human isolated islets as well as insulinoma cell lines (MIN6). Immunohistochemistry and subcellular localization were performed to localize the protein within the cell. Small interfering RNA knockdown of GLUT9 was used to determine the effect of this transporter, in the presence of GLUT2, on cell metabolism and insulin secretion in MIN6 and INS cells. In this report we demonstrate that GLUT9a and GLUT9b are expressed in pancreatic islets and that this expression localizes to insulin containing beta-cells. Subcellular localization studies indicate that mGLUT9b is found associated with the plasma membrane as well as in the high-density microsome fraction and low-density microsome fraction, whereas mGLUT9a appears to be located only in the high-density microsome and low-density microsome under basal conditions. Functionally GLUT9 appears to participate in the regulation of glucose-stimulated insulin secretion in addition to GLUT2. small interfering RNA knockdown of GLUT9 results in reduced cellular ATP levels that correlate with reductions in glucose-stimulated insulin secretion in MIN6 and INS cells. These studies confirm the expression of GLUT9a and GLUT9b in murine and human beta cells and suggest that GLUT9 may participate in glucose-sensing in beta-cells. PMID- 19808779 TI - Curcumin inhibits srebp-2 expression in activated hepatic stellate cells in vitro by reducing the activity of specificity protein-1. AB - Elevated levels of cholesterol/low-density lipoprotein (LDL) are a risk factor for the development of nonalcoholic steatohepatitis and its associated hepatic fibrosis. However, underlying mechanisms remain elusive. We previously reported that curcumin induced gene expression of peroxisome proliferator-activated receptor (PPAR)-gamma and stimulated its activity, leading to the inhibition of the activation of hepatic stellate cells (HSCs), the major effector cells during hepatic fibrogenesis. We recently showed that curcumin suppressed gene expression of LDL receptor in activated HSCs in vitro by repressing gene expression of the transcription factor sterol regulatory element binding protein-2 (SREBP-2), leading to the reduction in the level of intracellular cholesterol in HSCs and to the attenuation of the stimulatory effects of LDL on HSCs activation. The current study aimed at exploring molecular mechanisms by which curcumin inhibits srebp-2 expression in HSCs. Promoter deletion assays, mutagenesis assays, and EMSAs localize a specificity protein-1 (SP-1) binding GC-box in the srebp-2 promoter, which is responsible for enhancing the promoter activity and responding to curcumin in HSCs. Curcumin suppresses gene expression of SP-1 and reduces its trans-activation activity, which are mediated by the activation of PPARgamma. The inhibitory effect of curcumin on SP-1 binding to the GC-box is confirmed by chromatin immuno-precipitation. In summary, our results demonstrate that curcumin inhibits srebp-2 expression in cultured HSCs by activating PPARgamma and reducing the SP-1 activity, leading to the repression of ldlr expression. These results provide novel insights into molecular mechanisms by which curcumin inhibits LDL induced HSC activation. PMID- 19808780 TI - Implementation fidelity of packaged teen smoking cessation treatments delivered in community-based settings. AB - Efficacious 'packaged' teen smoking cessation treatment programs, those developed by national organizations, are widely disseminated to local communities to help teens quit smoking. The implementation fidelity of these programs in community settings has not been documented. The efficacy of these programs could be lessened if they are not implemented as intended. Data from Helping Young Smokers Quit describe the frequency and types of modifications made to packaged teen cessation treatment programs for community delivery. A national sample of 591 community-based teen tobacco cessation treatment programs was profiled and 59% used a single packaged treatment program. Bivariate analyses found that 63% of program administrators reported implementing their program as planned; 37% modified their selected program. The most frequently reported modifications were made to the length and format of the program. Of those who modified their programs, >90% reported multiple program modifications (e.g. length and content). Administrators modified their programs to accommodate implementation barriers, such as time constraints and low participant enrollment, and to address the needs of participants with multiple risk behaviors that are co-morbid with tobacco use. PMID- 19808781 TI - A mouse model offers novel insights into the myopathy and tendinopathy often associated with pseudoachondroplasia and multiple epiphyseal dysplasia. AB - Pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED) are relatively common skeletal dysplasias belonging to the same bone dysplasia family. PSACH is characterized by generalized epi-metaphyseal dysplasia, short limbed dwarfism, joint laxity and early onset osteoarthritis. MED is a milder disease with radiographic features often restricted to the epiphyses of the long bones. PSACH and some forms of MED result from mutations in cartilage oligomeric matrix protein (COMP), a pentameric glycoprotein found in cartilage, tendon, ligament and muscle. PSACH-MED patients often have a mild myopathy characterized by mildly increased plasma creatine kinase levels, a variation in myofibre size and/or small atrophic fibres. In some instances, patients are referred to neuromuscular clinics prior to the diagnosis of an underlying skeletal dysplasia; however, the myopathy associated with PSACH-MED has not previously been studied. In this study, we present a detailed study of skeletal muscle, tendon and ligament from a mouse model of mild PSACH harbouring a COMP mutation. Mutant mice exhibited a progressive muscle weakness associated with an increased number of muscle fibres with central nuclei at the perimysium and at the myotendinous junction. Furthermore, the distribution of collagen fibril diameters in the mutant tendons and ligaments was altered towards thicker collagen fibrils, and the tendons became more lax in cyclic strain tests. We hypothesize that the myopathy in PSACH-MED originates from an underlying tendon and ligament pathology that is a direct result of structural abnormalities to the collagen fibril architecture. This is the first comprehensive characterization of the musculoskeletal phenotype of PSACH-MED and is directly relevant to the clinical management of these patients. PMID- 19808787 TI - Genetics of human neural tube defects. AB - Neural tube defects (NTDs) are common, severe congenital malformations whose causation involves multiple genes and environmental factors. Although more than 200 genes are known to cause NTDs in mice, there has been rather limited progress in delineating the molecular basis underlying most human NTDs. Numerous genetic studies have been carried out to investigate candidate genes in cohorts of patients, with particular reference to those that participate in folate one carbon metabolism. Although the homocysteine remethylation gene MTHFR has emerged as a risk factor in some human populations, few other consistent findings have resulted from this approach. Similarly, attention focused on the human homologues of mouse NTD genes has contributed only limited positive findings to date, although an emerging association between genes of the non-canonical Wnt (planar cell polarity) pathway and NTDs provides candidates for future studies. Priorities for the next phase of this research include: (i) larger studies that are sufficiently powered to detect significant associations with relatively minor risk factors; (ii) analysis of multiple candidate genes in groups of well genotyped individuals to detect possible gene-gene interactions; (iii) use of high throughput genomic technology to evaluate the role of copy number variants and to detect 'private' and regulatory mutations, neither of which have been studied to date; (iv) detailed analysis of patient samples stratified by phenotype to enable, for example, hypothesis-driven testing of candidates genes in groups of NTDs with specific defects of folate metabolism, or in groups of fetuses with well-defined phenotypes such as craniorachischisis. PMID- 19808788 TI - Aicardi-Goutieres syndrome and related phenotypes: linking nucleic acid metabolism with autoimmunity. AB - Aicardi-Goutieres syndrome (AGS) is a genetically determined encephalopathy demonstrating phenotypic overlap both with the sequelae of congenital infection and with systemic lupus erythematosus (SLE). Recent molecular advances have revealed that AGS can be caused by mutations in any one of five genes, most commonly on a recessive basis but occasionally as a dominant trait. Like AGS, SLE is associated with a perturbation of type I interferon metabolism. Interestingly then, heterozygous mutations in the AGS1 gene TREX1 underlie a cutaneous subtype of SLE-called familial chilblain lupus, and mutations in TREX1 represent the single most common cause of monogenic SLE identified to date. Evidence is emerging to show that the nucleases defective in AGS are involved in removing endogenously produced nucleic acid (NA) species, and that a failure of this removal results in activation of the immune system. This hypothesis explains the phenotypic overlap of AGS with congenital infection and some aspects of SLE, where an equivalent type I interferon-mediated innate immune response is triggered by viral and self NAs, respectively. The combined efforts of clinicians, geneticists, immunologists and cell biologists are producing rapid progress in the understanding of AGS and overlapping autoimmune disorders. These studies provide important insights into the pathogenesis of SLE and beg urgent questions about the development and use of immunosuppressive therapies in AGS and related phenotypes. PMID- 19808789 TI - Genome-wide association studies in Alzheimer's disease. AB - Genome-wide association studies (GWAS) have gained considerable momentum over the last couple of years for the identification of novel complex disease genes. In the field of Alzheimer's disease (AD), there are currently eight published and two provisionally reported GWAS, highlighting over two dozen novel potential susceptibility loci beyond the well-established APOE association. On the basis of the data available at the time of this writing, the most compelling novel GWAS signal has been observed in GAB2 (GRB2-associated binding protein 2), followed by less consistently replicated signals in galanin-like peptide (GALP), piggyBac transposable element derived 1 (PGBD1), tyrosine kinase, non-receptor 1 (TNK1). Furthermore, consistent replication has been recently announced for CLU (clusterin, also known as apolipoprotein J). Finally, there are at least three replicated loci in hitherto uncharacterized genomic intervals on chromosomes 14q32.13, 14q31.2 and 6q24.1 likely implicating the existence of novel AD genes in these regions. In this review, we will discuss the characteristics and potential relevance to pathogenesis of the outcomes of all currently available GWAS in AD. A particular emphasis will be laid on findings with independent data in favor of the original association. PMID- 19808790 TI - Thermosensory and mechanosensory perception in human genetic disease. AB - Peripheral sensory perception is established through an elaborate network of specialized neurons that mediate the translation of extraorganismal stimuli through the use of a broad array of receptors and downstream effector molecules. Studies of human genetic disorders, as well as mouse and other animal models, have identified some of the key molecules necessary for peripheral innervation and function. These findings have, in turn, yielded new insights into the developmental networks and homeostatic mechanisms necessary for the transformation of external stimuli into interpretable electrical impulses. In this review, we will summarize and discuss some of the genes/proteins implicated in two particular aspects of sensory perception, thermosensation and mechanosensation, highlighting pathways whose perturbation leads to both isolated and syndromic sensory deficits. PMID- 19808791 TI - Mutations in TDP-43 link glycine-rich domain functions to amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is the most common adult motor neuron disease that affects approximately 2/100,000 individuals each year worldwide. Patients with ALS suffer from rapidly progressive degeneration of motor neurons ultimately leading to death. The major pathological features observed in post-mortem tissue from patients with ALS are motor neuron loss, cortical spinal tract degeneration, gliosis and cytoplasmic neuronal inclusions formed by TDP-43 or TAR DNA binding Protein with a molecular mass of 43 kDa, which are now recognized as the signature lesions of sporadic ALS. TDP-43 possesses two RNA binding domains (RBD) and a glycine-rich C terminus classifying it with other heterogeneous nuclear ribonucleoproteins known as 2XRBD-Gly proteins. A number of reports showed that a subset of patients with ALS possess mutations in the TDP-43 (TARDBP) gene. This further strengthens the hypotheses that gain of toxic function or loss of function in TDP-43 causes ALS. Currently, 29 different TARDBP missense mutations have been reported in 51 unrelated sporadic or familial ALS cases and two cases of ALS plus concomitant frontotemporal lobar degeneration with a remarkable concentration of mutations in the C-terminal glycine-rich domain of TDP-43. As these mutations will most certainly be an invaluable tool for the design and implementation of ALS animal and cell models, as well as serve as a platform for exploring the pathobiology of TDP-43, here we summarize the identified pathogenic TARDBP mutations and their potential impact on our understanding of the role of TDP-43 in disease. PMID- 19808792 TI - Cancer genome sequencing: a review. AB - A genomic era of cancer studies is developing rapidly, fueled by the emergence of next-generation sequencing technologies that provide exquisite sensitivity and resolution. This article discusses several areas within cancer genomics that are being transformed by the application of new technology, and in the process are dramatically expanding our understanding of this disease. Although, we anticipate that there will be many exciting discoveries in the near future, the ultimate success of these endeavors rests on our ability to translate what is learned into better diagnosis, treatment and prevention of cancer. PMID- 19808793 TI - Mitochondrial dynamics--fusion, fission, movement, and mitophagy--in neurodegenerative diseases. AB - Neurons are metabolically active cells with high energy demands at locations distant from the cell body. As a result, these cells are particularly dependent on mitochondrial function, as reflected by the observation that diseases of mitochondrial dysfunction often have a neurodegenerative component. Recent discoveries have highlighted that neurons are reliant particularly on the dynamic properties of mitochondria. Mitochondria are dynamic organelles by several criteria. They engage in repeated cycles of fusion and fission, which serve to intermix the lipids and contents of a population of mitochondria. In addition, mitochondria are actively recruited to subcellular sites, such as the axonal and dendritic processes of neurons. Finally, the quality of a mitochondrial population is maintained through mitophagy, a form of autophagy in which defective mitochondria are selectively degraded. We review the general features of mitochondrial dynamics, incorporating recent findings on mitochondrial fusion, fission, transport and mitophagy. Defects in these key features are associated with neurodegenerative disease. Charcot-Marie-Tooth type 2A, a peripheral neuropathy, and dominant optic atrophy, an inherited optic neuropathy, result from a primary deficiency of mitochondrial fusion. Moreover, several major neurodegenerative diseases--including Parkinson's, Alzheimer's and Huntington's disease--involve disruption of mitochondrial dynamics. Remarkably, in several disease models, the manipulation of mitochondrial fusion or fission can partially rescue disease phenotypes. We review how mitochondrial dynamics is altered in these neurodegenerative diseases and discuss the reciprocal interactions between mitochondrial fusion, fission, transport and mitophagy. PMID- 19808794 TI - Recent insights into the pathogenesis of hyperuricaemia and gout. AB - Gout is a common rheumatic disease in humans which is characterized by elevation in serum uric acid levels, and deposition of uric acid crystals in the joint. Hyperuricaemia is the primary risk factor for the development of gout and primates have uniquely high levels of serum uric acid due to missense mutations in the uricase gene. Levels of serum uric acid are known to be highly heritable, and mutations in genes which encode enzymes in the purine salvage pathway have long been recognized as rare causes of gout. Until recently, however, little has been known about the genetic determinants of urate metabolism and susceptibility to gout in the general population. Over recent months, a series of large scale genome wide association studies have been performed which have shed new light on the genes which regulate serum uric acid levels and susceptibility to gout. Most of these genes seem to be involved in regulating the renal excretion of uric acid which underscores the importance of reduced urate excretion as opposed to increased endogenous production as a cause of gout. Further work will now be required to investigate the mechanisms by which these genetic variants regulate urate excretion and serum urate levels. However, it seems likely that the genes so far identified will represent new molecular targets for the design of drugs to enhance urate excretion and the genetic variants that predispose to gout might be of value as genetic markers of susceptibility to gout. PMID- 19808795 TI - Genetics of nephrotic syndrome: connecting molecular genetics to podocyte physiology. AB - Urinary losses of macromolecules in nephrotic syndrome (NS) reflect a dysfunction of the highly permselective glomerular filtration barrier. Genetic studies of hereditary forms of NS have led to the identification of proteins playing a crucial role in slit-diaphragm signalling, regulation of actin cytoskeleton dynamics, maintenance of podocyte integrity and cell-matrix interactions. This review will focus on recent molecular and clinical findings in the field of genetics of NS, thereby providing a better understanding of the complex glomerular filtration barrier physiology. PMID- 19808796 TI - Predictive chromatin signatures in the mammalian genome. AB - The DNA sequence of an organism is a blueprint of life: it harbors not only the information about proteins and other molecules produced in each cell, but also instructions on when and where such molecules are made. Chromatin, the structure of histone and DNA that has co-evolved with eukaryotic genome, also contains information that indicates the function and activity of the underlying DNA sequences. Such information exists in the form of covalent modifications to the histone proteins that comprise the nucleosome. Thanks to the development of high throughput technologies such as DNA microarrays and next generation DNA sequencing, we have begun to associate the various combinations of chromatin modification patterns with functional sequences in the human genome. Here, we review the rapid progress from descriptive observations of histone modification profiles to highly predictive models enabling use of chromatin signatures to enumerate novel functional sequences in mammalian genomes that have escaped previous detection. PMID- 19808797 TI - Transgenerational genetic effects on phenotypic variation and disease risk. AB - Traditionally, we understand that individual phenotypes result primarily from inherited genetic variants together with environmental exposures. However, many studies showed that a remarkable variety of factors including environmental agents, parental behaviors, maternal physiology, xenobiotics, nutritional supplements and others lead to epigenetic changes that can be transmitted to subsequent generations without continued exposure. Recent discoveries show transgenerational epistasis and transgenerational genetic effects where genetic factors in one generation affect phenotypes in subsequent generation without inheritance of the genetic variant in the parents. Together these discoveries implicate a key signaling pathway, chromatin remodeling, methylation, RNA editing and microRNA biology. This exceptional mode of inheritance complicates the search for disease genes and represents perhaps an adaptation to transmit useful gene expression profiles from one generation to the next. In this review, I present evidence for these transgenerational genetic effects, identify their common features, propose a heuristic model to guide the search for mechanisms, discuss the implications, and pose questions whose answers will begin to reveal the underlying mechanisms. PMID- 19808798 TI - The resolution of the genetics of gene expression. AB - Understanding the influence of genetics on the molecular mechanisms underpinning human phenotypic diversity is fundamental to being able to predict health outcomes and treat disease. To interrogate the role of genetics on cellular state and function, gene expression has been extensively used. Past and present studies have highlighted important patterns of heritability, population differentiation and tissue-specificity in gene expression. Current and future studies are taking advantage of systems biology-based approaches and advances in sequencing technology: new methodology aims to translate regulatory networks to enrich pathways responsible for disease etiology and 2nd generation sequencing now offers single-molecular resolution of the transcriptome providing unprecedented information on the structural and genetic characteristics of gene expression. Such advances are leading to a future where rich cellular phenotypes will facilitate understanding of the transmission of genetic effect from the gene to organism. PMID- 19808799 TI - Control of fetal hemoglobin: new insights emerging from genomics and clinical implications. AB - Increased levels of fetal hemoglobin (HbF, alpha(2)gamma(2)) are of no consequence in healthy adults, but confer major clinical benefits in patients with sickle cell anemia (SCA) and beta thalassemia, diseases that represent major public health problems. Inter-individual HbF variation is largely genetically controlled, with one extreme caused by mutations involving the beta globin gene (HBB) complex, historically referred to as pancellular hereditary persistence of fetal hemoglobin (HPFH). These Mendelian forms of HPFH are rare and do not explain the common form of heterocellular HPFH which represents the upper tail of normal HbF variation, and is clearly inherited as a quantitative genetic trait. Genetic studies have identified three major quantitative trait loci (QTLs) (Xmn1 HBG2, HBS1L-MYB intergenic region on chromosome 6q23, and BCL11A on chromosome 2p16) that account for 20-50% of the common variation in HbF levels in patients with SCA and beta thalassemia, and in healthy adults. Two of the major QTLs include oncogenes, emphasizing the importance of cell proliferation and differentiation as an important contribution to the HbF phenotype. The review traces the story of HbF quantitative genetics that uncannily mirrors the changing focus in genetic methodology, from candidate genes through positional cloning, to genome-wide association, that have expedited the dissection of the genetic architecture underlying HbF variability. These genetic results have already provided remarkable insights into molecular mechanisms that underlie the hemoglobin 'switch'. PMID- 19808800 TI - Trichothiodystrophy view from the molecular basis of DNA repair/transcription factor TFIIH. AB - Trichothiodystrophy (TTD) is a rare autosomal recessive disorder characterized by brittle hair and also associated with various systemic symptoms. Approximately half of TTD patients exhibit photosensitivity, resulting from the defect in the nucleotide excision repair. Photosensitive TTD is due to mutations in three genes encoding XPB, XPD and p8/TTDA subunits of the DNA repair/transcription factor TFIIH. Mutations in these subunits disturb either the catalytic and/or the regulatory activity of the two XPB, XPD helicase/ATPases and consequently are defective in both, DNA repair and transcription. Moreover, mutations in any of these three TFIIH subunits also disturb the overall architecture of the TFIIH complex and its ability to transactivate certain nuclear receptor-responsive genes, explaining in part, some of the TTD phenotypes. PMID- 19808805 TI - MYB83 is a direct target of SND1 and acts redundantly with MYB46 in the regulation of secondary cell wall biosynthesis in Arabidopsis. AB - It has been proposed that the transcriptional regulation of secondary wall biosynthesis in Arabidopsis is controlled by a transcriptional network mediated by SND1 and its close homologs. Uncovering all the transcription factors and deciphering their interrelationships in the network are essential for our understanding of the molecular mechanisms underlying the transcriptional regulation of biosynthesis of secondary walls, the major constituent of wood and fibers. Here, we present functional evidence that the MYB83 transcription factor is another molecular switch in the SND1-mediated transcriptional network regulating secondary wall biosynthesis. MYB83 is specifically expressed in fibers and vessels where secondary wall thickening occurs. Its expression is directly activated by SND1 and its close homologs, including NST1, NST2, VND6 and VND7, indicating that MYB83 is their direct target. MYB83 overexpression is able to activate a number of the biosynthetic genes of cellulose, xylan and lignin and concomitantly induce ectopic secondary wall deposition. In addition, its overexpression upregulates the expression of several transcription factors involved in regulation of secondary wall biosynthesis. Dominant repression of MYB83 functions or simultaneous RNAi inhibition of MYB83 and MYB46 results in a reduction in secondary wall thickening in fibers and vessels and a deformation of vessels. Furthermore, double T-DNA knockout mutations of MYB83 and MYB46 cause a lack of secondary walls in vessels and an arrest in plant growth. Together, these results demonstrate that MYB83 and MYB46, both of which are SND1 direct targets, function redundantly in the transcriptional regulatory cascade leading to secondary wall formation in fibers and vessels. PMID- 19808806 TI - Ectopic overexpression of the transcription factor OsGLK1 induces chloroplast development in non-green rice cells. AB - For systematic and genome-wide analyses of rice gene functions, we took advantage of the full-length cDNA overexpresser (FOX) gene-hunting system and generated >12 000 independent FOX-rice lines from >25 000 rice calli treated with the rice-FOX Agrobacterium library. We found two FOX-rice lines generating green calli on a callus-inducing medium containing 2,4-D, on which wild-type rice calli became ivory yellow. In both lines, OsGLK1 cDNA encoding a GARP transcription factor was ectopically overexpressed. Using rice expression-microarray and northern blot analyses, we found that a large number of nucleus-encoded genes involved in chloroplast functions were highly expressed and transcripts of plastid-encoded genes, psaA, psbA and rbcL, increased in the OsGLK1-FOX calli. Transmission electron microscopy showed the existence of differentiated chloroplasts with grana stacks in OsGLK1-FOX calli cells. However, in darkness, OsGLK1-FOX calli did not show a green color or develop grana stacks. Furthermore, we found developed chloroplasts in vascular bundle and bundle sheath cells of coleoptiles and leaves from OsGLK1-FOX seedlings. The OsGLK1-FOX calli exhibited high photosynthetic activity and were able to grow on sucrose-depleted media, indicating that developed chloroplasts in OsGLK1-FOX rice calli are functional and active. We also observed that the endogenous OsGLK1 mRNA level increased synchronously with the greening of wild-type calli after transfer to plantlet regeneration medium. These results strongly suggest that OsGLK1 regulates chloroplast development under the control of light and phytohormones, and that it is a key regulator of chloroplast development. PMID- 19808807 TI - High temperatures cause male sterility in rice plants with transcriptional alterations during pollen development. AB - Plant male reproductive development is highly organized and sensitive to various environmental stressors, including high temperature. We have established an experimental procedure to evaluate high temperature injury in japonica rice plants. High temperature treatment (39 degrees C/30 degrees C) starting at the microspore stage repeatedly reduced spikelet fertility in our system. Morphological observations revealed that pollen viability in plants exposed to high temperatures was lower than that in control plants. Most pollen grains in high temperature-treated plants displayed a normal round shape and stained reddish purple with Alexander's reagent; however, the pollen grains were very poorly attached and displayed limited germination on the stigma. To investigate gene regulatory mechanisms in the anther in high temperature environments, DNA microarray analysis was performed by comparing non-treated samples with samples treated with 2-4 d of high heat. Genes responsive to high temperatures were identified from clustering of microarray data. Among these, at least 13 were designated as high temperature-repressed genes in the anther. Expression analyses revealed that these genes were expressed specifically in the immature anther mainly in the tapetum at the microspore stage and down-regulated after 1 d of high temperature. The expression levels of Osc6, OsRAFTIN and TDR, which are tapetum-specific genes, were unaffected by high temperatures. These results suggest that not all tapetal genes are inhibited by increased temperatures and the tapetum itself is not degraded in such an environment. However, high temperatures may disrupt some of the tapetum functions required for pollen adhesion and germination on the stigma. PMID- 19808808 TI - RESOPS: a database for analyzing the correspondence of RNA editing sites to protein three-dimensional structures. AB - Transcripts from mitochondrial and chloroplast DNA of land plants often undergo cytidine to uridine conversion-type RNA editing events. RESOPS is a newly built database that specializes in displaying RNA editing sites of land plant organelles on protein three-dimensional (3D) structures to help elucidate the mechanisms of RNA editing for gene expression regulation. RESOPS contains the following information: unedited and edited cDNA sequences with notes for the target nucleotides of RNA editing, conceptual translation from the edited cDNA sequence in pseudo-UniProt format, a list of proteins under the influence of RNA editing, multiple amino acid sequence alignments of edited proteins, the location of amino acid residues coded by codons under the influence of RNA editing in protein 3D structures and the statistics of biased distributions of the edited residues with respect to protein structures. Most of the data processing procedures are automated; hence, it is easy to keep abreast of updated genome and protein 3D structural data. In the RESOPS database, we clarified that the locations of residues switched by RNA editing are significantly biased to protein structural cores. The integration of different types of data in the database also help advance the understanding of RNA editing mechanisms. RESOPS is accessible at http://cib.cf.ocha.ac.jp/RNAEDITING/. PMID- 19808809 TI - Arabidopsis OPT6 is an oligopeptide transporter with exceptionally broad substrate specificity. AB - Oligopeptide transporters (OPTs) are found in fungi, bacteria and plants. The nine Arabidopsis thaliana OPT genes are expressed mainly in the vasculature and are thought to transport tetra- and pentapeptides, and peptide-like substrates such as glutathione. Expression of AtOPT6 in Xenopus laevis oocytes demonstrated that AtOPT6 transports many tetra- and pentapeptides. In addition, AtOPT6 transported reduced glutathione (GSH), a tripeptide, but not oxidized glutathione (GSSG). Recent data showed that Candida albicans OPTs can transport peptides up to eight amino acids in length. AtOPT6 transported mammalian signaling peptides up to 10 amino acids in length and, in addition, known plant development- and nematode pathogenesis-associated peptides up to 13 amino acids long. AtOPT6 displayed high affinity for penta- and dodecapeptides, but low affinity for GSH. In comparison the Saccharomyces cerevisiae ScOPT1 was incapable of transporting any of the longer peptides tested. These data demonstrate the necessity of experimentally determining substrate specificity of individual OPTs, and lay a foundation for structure/function studies. Characterization of the AtOPT6 substrate range provides a basis for investigating the possible physiological function of AtOPT6 in peptide signaling and thiol transport in response to stress. PMID- 19808810 TI - Rapid emotion regulation after mood induction: age and individual differences. AB - Previous research has suggested that emotion regulation improves with age. This study examined both age and individual differences in online emotion regulation after a negative mood induction. We found evidence that older adults were more likely to rapidly regulate their emotions than were younger adults. Moreover, older adults who rapidly regulated had lower trait anxiety and depressive symptoms and higher levels of optimism than their same-age peers who did not rapidly regulate. Measuring mood change over an extended time revealed that older rapid regulators still reported increased levels of positive affect over 20 min later, whereas young adult rapid regulators' moods had declined. These results highlight the importance of considering individual differences when examining age differences in online emotion regulation. PMID- 19808811 TI - Reliability of patient responses in pay for performance schemes: analysis of national General Practitioner Patient Survey data in England. AB - OBJECTIVE: To assess the robustness of patient responses to a new national survey of patient experience as a basis for providing financial incentives to doctors. DESIGN: Analysis of the representativeness of the respondents to the GP Patient Survey compared with those who were sampled (5.5 million patients registered with 8273 general practices in England in January 2009) and with the general population. Analysis of non-response bias looked at the relation between practice response rates and scores on the survey. Analysis of the reliability of the survey estimated the proportion of the variance of practice scores attributable to true differences between practices. RESULTS: The overall response rate was 38.2% (2.2 million responses), which is comparable to that in surveys using similar methodology in the UK. Men, young adults, and people living in deprived areas were under-represented among respondents. However, for questions related to pay for performance, there was no systematic association between response rates and questionnaire scores. Two questions which triggered payments to general practitioners were reliable measures of practice performance, with average practice-level reliability coefficients of 93.2% and 95.0%. Less than 3% and 0.5% of practices had fewer than the number of responses required to achieve conventional reliability levels of 90% and 70%. A change to the payment formula in 2009 resulted in an increase in the average impact of random variation in patient scores on payments to general practitioners compared with payments made in 2007 and 2008. CONCLUSIONS: There is little evidence to support the concern of some general practitioners that low response rates and selective non-response bias have led to systematic unfairness in payments attached to questionnaire scores. The study raises issues relating to the validity and reliability of payments based on patient surveys and provides lessons for the UK and for other countries considering the use of patient experience as part of pay for performance schemes. PMID- 19808812 TI - Asbestos and the lung. Diffuse pleural thickening has new definition for claims. PMID- 19808813 TI - Ovarian cancer. Usefulness of abdominal symptoms in early diagnosis. PMID- 19808814 TI - Symptoms are not early signs of ovarian cancer. PMID- 19808815 TI - Bran and irritable bowels. Bran's deleterious effects: much ado about nothing. PMID- 19808816 TI - Rosiglitazone and pioglitazone. Beware fractures. PMID- 19808817 TI - A/H1N1 flu pandemic. A/H1N1 and other viruses affecting cystic fibrosis. PMID- 19808819 TI - A/H1N1 flu pandemic. Managing neutropenic sepsis. PMID- 19808820 TI - Sarcoidosis. Technique to enable diagnosis. PMID- 19808821 TI - From sick notes to fit notes. Bridging the chasm. PMID- 19808822 TI - From sick notes to fit notes. Not a simple answer. PMID- 19808823 TI - Interventions and knee pain. When knee pain is not osteoarthritis. PMID- 19808826 TI - GPs are to refer suspected cancer patients for tests in a week under new scheme. PMID- 19808827 TI - Drug prevention of hypertension. Contradicting Cochrane. PMID- 19808829 TI - Ethics in sports medicine. PMID- 19808830 TI - Improving the use of IT in the NHS. PMID- 19808831 TI - The direct and indirect costs of explosive violence. PMID- 19808835 TI - European agency approves swine flu vaccines for licensing. PMID- 19808837 TI - Surgery-induced inflammation in relation to age, muscle endurance, and self perceived fatigue. AB - BACKGROUND: Elective abdominal surgery can be considered as a model for an important acute inflammatory trigger in human participants. The aim of the study was to explore the effect of surgery-induced inflammation on muscle strength, endurance, and self-perceived fatigue and its relation with age. METHODS: Sixty six elective abdominal surgery patients aged 24-91 years were assessed before and at the second and fourth day after surgical intervention. Outcome parameters were grip strength, muscle endurance, fatigue subscale of the Profile of Mood State and visual analog scale for pain, and the circulating inflammatory mediators C reactive protein, interleukin (IL)-6, and tumor necrosis factor-alpha (TNF alpha). RESULTS: All parameters worsened postoperatively (p < .01) and remained significantly (p < .05) worse until the fourth day postsurgery, except for TNF alpha (no significant change). Older age was related to higher surgery-induced IL 6 levels at the second (p < .05) and fourth postoperative (p < .01) day and to worse self-perceived fatigue and muscle endurance (both p < .05) at the fourth postoperative day. Higher pain levels at the second day following surgery was related to more self-perceived fatigue (p < .05). Worsening muscle endurance following surgery was significantly related to higher IL-6 release following surgery (p < .01) and self-perceived fatigue (p < .05) at the fourth day following the intervention. Age and surgery-induced increase in circulating IL-6 at Day 4 postsurgery was highest in patients showing both worsened muscle endurance and self-perceived fatigue (p < .05). CONCLUSIONS: Surgery-induced inflammation is related to reduced muscle endurance and the sensation of fatigue. Elderly patients suffer from a higher impact of surgery on muscle endurance. PMID- 19808838 TI - Effects of chronic overload on muscle hypertrophy and mTOR signaling in young adult and aged rats. AB - We examined the effect of 28 days of overload on mammalian target of rapamycin (mTOR) and extracellular signal-regulated kinase (ERK) signaling in young adult (Y; 6-month old) and aged (O; 30-month old) Fischer 344 x Brown Norway rats subjected to bilateral synergist ablation (SA) of two thirds of the gastrocnemius muscle or sham surgery (CON). Although plantaris (PLA) muscle hypertrophy was attenuated by aging, mTOR phosphorylation was 44% and 35% greater in Y SA and O SA compared with CON (p = .038). Ribosomal protein S6 phosphorylation was 114% and 24% higher in Y SA and O SA compared with CON (p = .009). Eukaryotic initiation factor 2Bepsilon phosphorylation was 33% and 9% higher in Y SA and O SA compared with CON (p = .04). Translational signaling in young adult and aged plantaris muscle is equally responsive to chronic overload. PMID- 19808839 TI - A case of bone, lung, pleural and liver metastases from renal cell carcinoma which responded remarkably well to zoledronic acid monotherapy. AB - Herein, we report a rare case in which bisphosphonate zoledronic acid (ZA) effectively treated not only multiple bone metastases but also lung, pleural and liver metastases from renal cell carcinoma (RCC). Recently, ZA is used to treat skeletal-related events (SREs) such as bone pain caused by bone metastasis from many kinds of cancer. The patient in the present report had multiple bone metastases from RCC. Remarkable improvement of the bone metastasis was observed following treatment with ZA at a dosage of 4 mg administered once every 4 weeks. Moreover, lung, pleural and liver metastases also diminished markedly in size in response to the treatment. The metastases have shown no progression for 20 months since starting the ZA treatment. We believe that the present report is the first of its kind announcing that ZA monotherapy has been effective for lung, pleural and liver metastases from RCC. PMID- 19808840 TI - Exercise, sex, menstrual cycle phase, and 17beta-estradiol influence metabolism related genes in human skeletal muscle. AB - Higher fat and lower carbohydrate and amino acid oxidation are observed in women compared with men during endurance exercise. We hypothesized that the observed sex difference is due to estrogen and that menstrual cycle phase or supplementation of men with 17beta-estradiol (E(2)) would coordinately influence the mRNA content of genes involved in lipid and/or carbohydrate metabolism in skeletal muscle. Twelve men and twelve women had muscle biopsies taken before and immediately after 90 min of cycling at 65% peak oxygen consumption (Vo(2peak)). Women were studied in the midfollicular (Fol) and midluteal (Lut) phases, and men were studied after 8 days of E(2) or placebo supplementation. Targeted RT-PCR was used to compare mRNA content for genes involved in transcriptional regulation and lipid, carbohydrate, and amino acid metabolism. Sex was the greatest predictor of substrate metabolism gene content. Sex affected the mRNA content of FATm, FABPc, SREBP-1c, mtGPAT, PPARdelta, PPARalpha, CPTI, TFP-alpha, GLUT4, HKII, PFK, and BCOADK (P < 0.05). E(2) administration significantly (P < 0.05) affected the mRNA content of PGC-1alpha, PPARalpha, PPARdelta, TFP-alpha, CPTI, SREBP-1c, mtGPAT, GLUT4, GS-1, and AST. Acute exercise increased the mRNA abundance for PGC-1alpha, HSL, FABPc, CPTI, GLUT4, HKII, and AST (P < 0.05). Menstrual cycle had a small effect on PPARdelta, GP, and glycogenin mRNA content. Overall, women have greater mRNA content for several genes involved in lipid metabolism, which is partially due to an effect of E(2). PMID- 19808841 TI - Acceptability and feasibility results of a strength-based skills training program for dementia caregiving dyads. AB - PURPOSE: The current article provides an in-depth description of a dyadic intervention for individuals with dementia and their family caregivers. Using a strength-based approach, caregiving dyads received skills training across 5 key areas: (a) education regarding dementia and memory loss, (b) effective communication, (c) managing memory loss, (d) staying active, and (e) recognizing emotions and behaviors. Results of the acceptability and feasibility of the intervention protocols are also presented. DESIGN AND METHODS: Caregiving dyads were randomly assigned to participate in the intervention. Participants in the treatment condition were asked to complete a series of evaluation questions after each intervention session and an overall evaluation of the program. Data were also collected from the intervention specialists who implemented the protocols. RESULTS: Overall, the evaluation data indicated that the content and process of the intervention were viewed as highly acceptable and feasible by both participants and intervention specialists. IMPLICATIONS: This article highlights the merit of using a strength-based approach for working with caregiving dyads with dementia and how a single intervention protocol can be used to address the goals of both care partners. Furthermore, the intervention program was found to be highly acceptable and feasible, which is an important aspect of developing dyadic protocols. PMID- 19808842 TI - The revised direct assessment of functional status for independent older adults. AB - PURPOSE: The original version of the Direct Assessment of Functional Status (DAFS), a measure of instrumental activities of daily living (IADLs), was found to have a ceiling effect in older adults living independently in the community. This suggested that the tasks measured, although relevant, do not require full use of this population's abilities, and thus, the instrument may not be sensitive to the early decrements in IADLs that can signal initial cognitive impairment and may not detect improvements in IADLs over time, which is especially important in intervention research. DESIGN AND METHODS: By removing items with little to no variation and adding more difficult subscales that emphasized medication management skills, we designed the DAFS-Extended (Direct Assessment of Functional Status-Revised [DAFS-R]) to be more challenging for elders living independently. RESULTS: Analysis with a sample of 45 older adults suggested that scores on the DAFS-R appear to be more normally distributed than on the original version. The DAFS was able to differentiate individuals with varying standard profile scores on the Rivermead Behavioural Memory Test memory performance instrument (normal, poor, and impaired). In addition, the reliability and validity of the DAFS-R were supported in this sample. IMPLICATIONS: Given the large number of older adults who regularly take multiple prescription medications, deficits in medication management skills can have serious consequences. A performance measure that emphasizes these higher level daily living skills can help providers screen for initial signs of functional decline. PMID- 19808843 TI - Does screening history explain the ethnic differences in stage at diagnosis of cervical cancer in New Zealand? AB - BACKGROUND: There are ethnic disparities in cervical cancer survival in New Zealand. The objectives of this study were to assess the associations of screening history, ethnicity, socio-economic status (SES) and rural residence with stage at diagnosis in women diagnosed with cervical cancer in New Zealand during 1994-2005. METHODS: The 2323 cases were categorized as 'ever screened' if they had had at least one smear prior to 6 months before diagnosis, and as 'regular screening' if they had had no more than 36 months between any two smears in the period 6-114 months before diagnosis. Logistic regression was used to estimate the associations of screening history, ethnicity, SES and urban/rural residence with stage at diagnosis. RESULTS: The percentages 'ever screened' were 43.3% overall, 24.8% in Pacific, 30.5% in Asian, 40.6% in Maori and 46.1% in 'Other' women. The corresponding estimates for 'regular screening' were 14.0, 5.7, 7.8, 12.5 and 15.3%. Women with 'regular screening' had a lower risk of late stage diagnosis [odds ratio (OR) 0.16, 95% confidence interval (CI) 0.10-0.26], and the effect was greater for squamous cell carcinoma (OR 0.12, 95% CI 0.07 0.23) than for adenocarcinoma (OR 0.32, 95% CI 0.13-0.82). The increased risk of late-stage diagnosis (OR 2.72, 95% CI 1.99-3.72) in Maori (compared with 'Other') women decreased only slightly when adjusted for screening history (OR 2.45, 95% CI 1.77-3.39). CONCLUSIONS: Over half of cases had not been 'ever screened'. Regular screening substantially lowered the risk of being diagnosed at a late stage. However, screening history does not appear to explain the ethnic differences in stage at diagnosis. PMID- 19808844 TI - Operational radiation protection in synchrotron light and free electron laser facilities. AB - The third-generation synchrotron radiation (SR) facilities are storage ring-based facilities with many insertion devices and photon beamlines, and have low injection beam power, but extremely high stored beam power. The fourth-generation X-ray free electron laser (FEL) facilities are based on an electron Linac with a long undulator and have high injection beam power. Due to its electron and photon beam characteristics and modes of operation, storage ring and photon beamlines have unique safety aspects, which are the main subjects of this paper. The shielding design limits, operational modes and beam losses are reviewed. Shielding analysis (source terms and methodologies) and interlocked safety systems for storage ring and photon beamlines (including SR and gas bremsstrahlung) are described. Specific safety issues for storage ring top-off injection and FEL facilities are discussed. Operational safety programme elements, e.g. operation authorisation, commissioning, training and radiation measurements, for SR facilities are also presented. PMID- 19808845 TI - Schwann Cell Migration Induced by Earthworm Extract via Activation of PAs and MMP2/9 Mediated through ERK1/2 and p38. AB - The earthworm, which has stasis removal and wound-healing functions, is a widely used Chinese herbal medicine in China. Schwann cell migration is critical for the regeneration of injured nerves. Schwann cells provide an essentially supportive activity for neuron regeneration. However, the molecular migration mechanisms induced by earthworms in Schwann cells remain unclear. Here, we investigate the roles of MAPK (ERK1/2, JNK and p38) pathways for earthworm-induced matrix degrading proteolytic enzyme (PAs and MMP2/9) production in Schwann cells. Moreover, earthworm induced phosphorylation of ERK1/2 and p38, but not JNK, activate the downstream signaling expression of PAs and MMPs in a time-dependent manner. Earthworm-stimulated ERK1/2 and p38 phosphorylation was attenuated by pretreatment with U0126 and SB203580, resulting in migration and uPA-related signal pathway inhibition. The results were confirmed using small interfering ERK1/2 and p38 RNA. These results demonstrated that earthworms can stimulate Schwann cell migration and up-regulate PAs and MMP2/9 expression mediated through the MAPK pathways, ERK1/2 and p38. Taken together, our data suggests the MAPKs (ERK1/2, p38)-, PAs (uPA, tPA)-, MMP (MMP2, MMP9) signaling pathway of Schwann cells regulated by earthworms might play a major role in Schwann cell migration and nerve regeneration. PMID- 19808846 TI - Excoecarianin, Isolated from Phyllanthus urinaria Linnea, Inhibits Herpes Simplex Virus Type 2 Infection through Inactivation of Viral Particles. AB - Phyllanthus urinaria Linnea (Euphorbiaceae) is one of the traditional medicinal plants widely used by oriental people to treat various diseases. We have previously demonstrated that the acetone extract of P. urinaria inhibits herpes simplex virus type 2 (HSV-2) but not HSV-1 infection. In a continuing effort to clarify the antiviral mechanisms of P. urinaria, we isolated the pure compound excoecarianin from the whole plant of P. urinaria through acetone extraction, and investigated its anti-HSV-1 and HSV-2 activities. Our results indicated that excoecarianin protected Vero cells from HSV-2 but not HSV-1 infection, and its 50% inhibitory concentration (IC(50)) was 1.4 +/- 0.1 MUM. The antiviral effective concentration of excoecarianin did not affect the viability or the morphology of Vero cells. Although excoecarianin inhibited HSV-2 infection, the inhibitory effect, however, was most prominent when excoecarianin was concurrently added with the virus. Pretreatment of Vero cells with excoecarianin with removal of the drug prior to infection did not yield any antiviral effects, and the same observation was made for post viral entry treatment. Subsequent studies revealed that excoecarianin inactivated HSV-2 virus particles to prevent viral infection. A synergistic antiviral effect against HSV-2 was also observed when Vero cells were treated with a combination of acyclovir (ACV) and excoecarianin. These results suggested that excoecarianin merits to be further explored as an entry inhibitor against HSV-2 and could potentially be investigated for combinatorial drug treatment with nucleoside analogues such as ACV in therapeutic management of HSV-2 infection. PMID- 19808847 TI - Maternal and fetal outcome in women with type 2 versus type 1 diabetes mellitus: a systematic review and metaanalysis. AB - CONTEXT: Glycemic disturbance is usually less severe in pregnant women with type 2 than in those with type 1 diabetes mellitus (DM). Nevertheless, a worse perinatal outcome in women with type 2 DM has been reported in some studies. OBJECTIVE: Our objective was to review maternal and fetal outcomes in pregnant women with type 2 vs. type 1 DM. STUDY SELECTION: We conducted a systematic review of papers providing original data on pregnancy outcomes in both type 2 and type 1 DM (Medline search of the period January 1, 1987, to June 30, 2008). Two independent investigators considered papers for eligibility, and a third one solved discrepancies. DATA EXTRACTION: Metaanalysis tools were used to compare four main outcomes (major congenital malformations, stillbirth, and neonatal and perinatal mortality) and 15 secondary ones (five maternal, 10 fetal). Meta analysis of Observational Studies in Epidemiology (MOOSE) guidelines were used to assess quality. DATA SYNTHESIS: Thirty-three studies qualified for inclusion of 3743 citations retrieved. Women with type 2 DM had lower glycated hemoglobin (HbA1c) at booking and throughout pregnancy but a higher risk of perinatal mortality [odds ratio (OR) 1.50, 95% confidence interval (CI) 1.15-1.96] without significant differences in the rates of major congenital malformations, stillbirth, and neonatal mortality. As to secondary outcomes, women with type 2 DM had less diabetic ketoacidosis (OR 0.09, 95% CI 0.02-0.34) and cesarean section (OR 0.80, 95% CI 0.59-0.94) without differences in other outcomes. CONCLUSIONS: Despite a milder glycemic disturbance, women with type 2 DM had no better perinatal outcomes than those with type 1, indicating that type 2 DM in pregnancy is a serious condition. PMID- 19808848 TI - ACTH deficiency, higher doses of hydrocortisone replacement, and radiotherapy are independent predictors of mortality in patients with acromegaly. AB - CONTEXT: A number of retrospective studies report that patients with acromegaly have increased morbidity and premature mortality, with standardized mortality ratios (SMR) of 1.3-3. Many patients with acromegaly develop hypopituitarism as a result of the pituitary adenoma itself or therapies such as surgery and radiotherapy. Pituitary radiotherapy and hypopituitarism have also been associated with an increased SMR. METHODS: Using the West MIDLANDS: Acromegaly database (n = 501; 275 female), we assessed the influence of prior radiotherapy and hypopituitarism (and replacement therapy) on mortality in patients with acromegaly. Median duration of follow-up was 14.0 yr (interquartile range, 7.9-21 yr). RESULTS: All-cause mortality was elevated [SMR, 1.7 (1.4, 2.0); P < 0.001]. On external analysis, prior radiotherapy, ACTH, and gonadotropin deficiency were associated with an elevated SMR [radiotherapy SMR, 2.1 (1.7-2.6); P = 0.006; ACTH deficiency SMR, 2.5 (1.9-3.2); P < 0.0005; and gonadotropin deficiency SMR, 2.1 (1.6-2.7); P = 0.037]. On internal analysis, the relative risk (RR) of mortality was increased in the radiotherapy [RR, 1.8 (1.2-2.8); P = 0.008] and ACTH deficiency groups [RR, 1.7 (1.2-2.5); P = 0.004], but not in the gonadotropin- or TSH-deficiency groups. In the ACTH-deficient group, increased replacement doses of hydrocortisone greater than 25 mg/d were associated with increased mortality compared to lower doses. CONCLUSIONS: Radiotherapy and ACTH deficiency are significantly associated with increased mortality in patients with acromegaly. In ACTH-deficient patients, a daily dose of more than 25 mg hydrocortisone is associated with increased mortality compared to lower doses. These results have important implications for the treatment of patients with acromegaly and also raise issues as to the optimum hydrocortisone treatment regimens for ACTH deficient patients. PMID- 19808849 TI - Aggregation of thyroid autoantibodies in twins from opposite-sex pairs suggests that microchimerism may play a role in the early stages of thyroid autoimmunity. AB - CONTEXT: Microchimerism is the presence of small populations of cells from one individual in another genetically distinct individual. This phenomenon can arise from pregnancy, blood transfusion, or bidirectional cell trafficking between twins in utero. Microchimerism has recently been proposed to play a role in the pathogenesis of thyroid autoimmunity. In that case, twins from opposite-sex pairs (OS) should have an increased risk of thyroid autoantibodies (TA). AIM: The aim of the study was to compare the frequency of TA in twin individuals from OS and monozygotic (MZ) twin pairs. DESIGN: This was a case-control study of 240 individuals (120 females and 120 males) from OS twin pairs (cases) and 568 control individuals from MZ pairs (284 females and 284 males). METHODS: Antibodies toward thyroid peroxidase (TPOAb), thyroglobulin (TgAb), and the TSH receptor (TSHRAb) were measured and considered positive if greater than 60 U/ml, greater than 60 U/ml, and greater than 1.0 U/liter, respectively. RESULTS: The frequency of TPOAb, TgAb, and TSHRAb among female cases was 15.0, 5.0, and 4.2%, respectively, which was higher than the corresponding prevalences in the female control population: 7.4% (P = 0.018), 1.1% (P = 0.023), and 0.7% (P = 0.026), respectively. However, when corrected for the number of phenotypes studied (TPOAb, TgAb, TSHRAb, and any thyroid antibody), the association remained significant only in the combined group, P(corrected) = 0.012. Essentially similar results were obtained in males. CONCLUSION: Both female and male twins from OS pairs, as opposed to MZ pairs, have an increased frequency of TA, indicating a potential role of microchimerism in developing TA. PMID- 19808850 TI - Influence of age and obesity on serum estradiol, estrone, and sex hormone binding globulin concentrations following oral estrogen administration in postmenopausal women. AB - OBJECTIVE: Hormone therapy (HT) increases the risk of venous thrombosis and stroke. Risk of venous thrombosis and stroke is higher in older, overweight, and obese women using HT. However, the impact of age and obesity on estrogen concentrations among HT users is not well defined. METHOD: We measured serum levels of estrone, total and free estradiol, and SHBG in 180 postmenopausal women participating in the Estrogen in the Prevention of Atherosclerosis Trial (EPAT), 91 receiving estradiol therapy (ET) and 89 taking placebo, every 6 months over 2 yr. Mean on-trial levels of estrogens and SHBG were compared across age, body mass index (BMI), and waist to hip ratio categories among ET users and placebo separately. RESULTS: Among the ET users, total (P = 0.01) and free estradiol (P = 0.002) were significantly directly associated with BMI adjusted for age. SHBG was inversely related to waist to hip ratio adjusted for age (P = 0.005). Age was not associated with any of the estrogen or SHBG concentrations in ET or placebo groups. BMI was positively associated with estrone concentrations among older but not younger ET users (P for interaction = 0.03). CONCLUSION: Overweight and obese women using ET attain greater concentrations of estrogen compared to women with normal BMI, whereas ET users with abdominal obesity attain lower SHBG levels. Obese older women using ET have the highest concentration of estrone. It may be useful to consider age and obesity when prescribing HT to minimize the risk of venous thrombosis or stroke in postmenopausal women. Further research regarding relationships among circulating hormone levels and risk for these conditions is required to substantiate this conclusion. PMID- 19808851 TI - Subclinical hypothyroidism in Korean preterm infants associated with high levels of iodine in breast milk. AB - CONTEXT: The dietary iodine intake of lactating women has been reported to be high in Korea. OBJECTIVES: The aim of this study was to assess iodine balance and to determine its relationship with thyroid function in preterm infants. DESIGN: Thyroid functions of preterm infants born at 34 wk gestation or less were evaluated in the first (n = 31) and third (n = 19) weeks. Mothers' breast milk (BM) and random urine samples of infants were taken on the same days for thyroid function tests. RESULTS: Iodine concentrations in BM were very high (198-8484 microg/liter), and one third of the infants had an iodine intake of more than 100 microg/kg per day at the third week after birth (excessive iodine intake group). At that time, the levels of TSH were positively correlated with urinary iodine (r = 0.622; P = 0.004). The frequencies of subclinical hypothyroidism were high in the excessive iodine intake group at the third and sixth weeks. The estimated daily iodine intake at the third week (51.2 +/- 45.5 vs. 149.0 +/- 103.8 microg/kg per day; P = 0.033), urinary iodine at the third week (913.2 +/- 1179.7 vs. 1651.3 +/- 1135.2 microg/liter; P = 0.051), and estimated daily iodine intake at the sixth week (32.8 +/- 35.5 vs. 92.1 +/- 51.2 microg/kg per day; P = 0.032) were significantly higher in infants with subclinical hypothyroidism than in controls. CONCLUSIONS: Excessive iodine intake from BM contributed to subclinical hypothyroidism in these preterm Korean infants. PMID- 19808852 TI - Glomerular filtration rate and parathyroid hormone secretion in primary hyperparathyroidism. AB - CONTEXT: The recent Third International Workshop on the Management of Asymptomatic Primary Hyperparathyroidism (PHPT) set 60 ml/min as the precise level of glomerular filtration rate (GFR) below which surgery is recommended because it is considered a threshold of concern in patients with PHPT. OBJECTIVE: The aim of the study was to investigate the relationship between different stages of renal insufficiency and PTH levels in PHPT patients. DESIGN: We conducted a cross-sectional study. PATIENTS AND METHODS: We studied 294 consecutive PHPT patients. Biochemical evaluation included total and ionized serum calcium, phosphate, creatinine, immunoreactive intact PTH, and 25-hydroxyvitamin D3 levels in the fasting state. GFR was assessed with the Modification of Diet in Renal Disease Study formula. RESULTS: The mean GFR of the whole group of PHPT patients was 92.3 +/- 31.6 ml/min x 1.73 m(2). The patients were divided into four groups according to National Kidney Foundation Disease Outcomes Quality Initiative (K/DOQI) guidelines: group 1 with normal or increased GRF (>90 ml/min x 1.73 m(2); n = 153); group 2 with mild decreased GFR (60-89 ml/min x 1.73 m(2); n = 90); group 3 with moderately decreased GFR (30-59 ml/min x 1.73 m(2); n = 45); and group 4 with severely decreased GFR (<30 ml/min x 1.73 m(2); n = 6). PTH levels were comparable across groups 1-3, whereas group 4 showed significantly higher PTH levels (P < 0.0001). CONCLUSION: In our series of PHPT patients, only a severe impairment of GFR was characterized by a further PTH increase. These findings challenge the concept of a PTH elevation below the threshold of 60 ml/min of GFR. PMID- 19808853 TI - The impact of exercise training compared to caloric restriction on hepatic and peripheral insulin resistance in obesity. AB - CONTEXT: It has been difficult to distinguish the independent effects of caloric restriction versus exercise training on insulin resistance. OBJECTIVE: Utilizing metabolic feeding and supervised exercise training, we examined the influence of caloric restriction vs. exercise training with and without weight loss on hepatic and peripheral insulin resistance. DESIGN, PARTICIPANTS, AND INTERVENTION: Thirty four obese, older subjects were randomized to: caloric restriction with weight loss (CR), exercise training with weight loss (EWL), exercise training without weight loss (EX), or controls. Based on an equivalent caloric deficit in EWL and CR, we induced matched weight loss. Subjects in the EX group received caloric compensation. Combined with [6,6(2)H(2)]glucose, an octreotide, glucagon, multistage insulin infusion was performed to determine suppression of glucose production (SGP) and insulin-stimulated glucose disposal (ISGD). Computed tomography scans were performed to assess changes in fat distribution. RESULTS: Body weight decreased similarly in EWL and CR, and did not change in EX and controls. The reduction in visceral fat was significantly greater in EWL (-71 +/- 15 cm(2)) compared to CR and EX. The increase in SGP was also almost 3-fold greater (27 +/- 2%) in EWL. EWL and CR promoted similar improvements in ISGD [+2.5 +/- 0.4 and 2.4 +/- 0.9 mg x kg fat-free mass (FFM)(-1) x min(-1)], respectively. CONCLUSIONS: EWL promoted the most significant reduction in visceral fat and the greatest improvement in SGP. Equivalent increases in ISGD were noted in EWL and CR, whereas EX provided a modest improvement. Based on our results, EWL promoted the optimal intervention-based changes in body fat distribution and systemic insulin resistance. PMID- 19808854 TI - Parasympathetic paragangliomas are part of the Von Hippel-Lindau syndrome. AB - CONTEXT: Von Hippel-Lindau (VHL) disease, caused by germline mutations in the VHL gene, is a hereditary tumor syndrome manifested by hemangioblastomas, clear cell renal cell carcinomas, and pheochromocytomas. In addition, a multitude of other rare tumors, including parasympathetic paragangliomas, can occur and even be the sole manifestation of VHL disease. The VHL gene is a bona fide tumor suppressor gene with biallelic inactivation contributing to tumor formation. However, in parasympathetic paragangliomas occurring in VHL disease, biallelic inactivation of the VHL gene has not been demonstrated to date. DESIGN: The head and neck paragangliomas of two VHL patients were analyzed for mutations by direct sequencing of the VHL gene. In addition loss of heterozygosity analysis was performed for three microsatellite loci near the VHL gene. To rule out other underlying genetic causes of the parasympathetic paragangliomas, mutation analysis of the SDHB, SDHC, and SDHD genes was also performed. RESULTS: Apart from germline VHL mutations, no additional mutations were found in the paraganglioma-related tumor suppressor genes SDHB, SDHC, and SDHD. Analysis of paraganglioma tissue revealed loss of the VHL wild-type allele in both tumors, indicating that in these tumors biallelic VHL gene inactivation occurred. CONCLUSIONS: These findings indicate that parasympathetic paragangliomas in VHL disease, although rare, are part of the syndrome and related to VHL gene inactivation. Clinicians should be aware of the potential occurrence of parasympathetic paragangliomas in VHL disease. PMID- 19808855 TI - Aerobic exercise increases peripheral and hepatic insulin sensitivity in sedentary adolescents. AB - CONTEXT: Data are limited on the effects of controlled aerobic exercise programs (without weight loss) on insulin sensitivity and glucose metabolism in children and adolescents. OBJECTIVE: To determine whether a controlled aerobic exercise program (without weight loss) improves peripheral and hepatic insulin sensitivity and affects glucose production (GPR), gluconeogenesis and glycogenolysis in sedentary lean and obese Hispanic adolescents. PATIENTS AND DESIGN: Twenty-nine post-pubertal adolescents (14 lean: 15.1 +/- 0.3 y; 20.6 +/- 0.8 kg/m(2); 18.9+/ 1.5% body fat and 15 obese: 15.6 +/- 0.4 y; 33.2 +/- 0.9 kg/m(2); 38.4 +/- 1.4% body fat) (mean +/- SE), completed a 12 wk aerobic exercise program (4 x 30 min/week at >or=70% of VO(2) peak). Peripheral and hepatic insulin sensitivity and glucose kinetics were quantified using GCMS pre- and post-exercise. RESULTS: No weight loss occurred. Lean and obese participants complied well with the program ( approximately 90% of the exercise sessions attended, resulting in approximately 15% increase in fitness in both groups). Peripheral and hepatic insulin sensitivity were higher in lean than obese adolescents but increased in both groups; peripheral insulin sensitivity by 35 +/- 14% (lean) (p < 0.05) and 59 +/- 19% (obese) (p < 0.01) and hepatic insulin sensitivity by 19 +/- 7% (lean) (p < 0.05) and 23 +/- 4% (obese) (p < 0.01). GPR, gluconeogenesis and glycogenolysis did not differ between the groups. GPR decreased slightly, 3 +/- 1% (lean) (p < 0.05) and 4 +/- 1% (obese) (p < 0.01). Gluconeogenesis remained unchanged, while glycogenolysis decreased slightly in the obese group (p < 0.01). CONCLUSION: This well accepted aerobic exercise program, without weight loss, is a promising strategy to improve peripheral and hepatic insulin sensitivity in lean and obese sedentary adolescents. The small decrease in GPR is probably of limited clinical relevance. PMID- 19808856 TI - Progesterone and mifepristone regulate L-type amino acid transporter 2 and 4F2 heavy chain expression in uterine leiomyoma cells. AB - CONTEXT: Progesterone and its receptor (PR) play key roles in uterine leiomyoma growth. Previously, using chromatin immunoprecipitation-based cloning, we uncovered L-type amino acid transporter 2 (LAT2) as a novel PR target gene. LAT2 forms heterodimeric complexes with 4F2 heavy chain (4F2hc), a single transmembrane domain protein essential for LAT2 to exert its function in the plasma membrane. Until now, little is known about the roles of LAT2/4F2hc in the regulation of the growth of human uterine leiomyoma. OBJECTIVE: The aim of the study is to investigate the regulation of LAT2 and 4F2hc by progesterone and the antiprogestin mifepristone and their functions in primary human uterine leiomyoma smooth muscle (LSM) cells and tissues from 39 premenopausal women. RESULTS: In primary LSM cells, progesterone significantly induced LAT2 mRNA levels, and this was blocked by cotreatment with mifepristone. Progesterone did not alter 4F2hc mRNA levels, whereas mifepristone significantly induced 4F2hc mRNA expression. Small interfering RNA knockdown of LAT2 or 4F2hc markedly increased LSM cell proliferation. LAT2, PR-B, and PR-A levels were significantly higher in freshly isolated LSM cells vs. adjacent myometrial cells. In vivo, mRNA levels of LAT2 and PR but not 4F2hc were significantly higher in leiomyoma tissues compared with matched myometrial tissues. CONCLUSION: We present evidence that progesterone and its antagonist mifepristone regulate the amino acid transporter system LAT2/4F2hc in leiomyoma tissues and cells. Our findings suggest that products of the LAT2/4F2hc genes may play important roles in leiomyoma cell proliferation. We speculate that critical ratios of LAT2 to 4F2hc regulate leiomyoma growth. PMID- 19808857 TI - MK2 regulates the early stages of skin tumor promotion. AB - The association between inflammation and tumorigenesis is well recognized. Mitogen-activated protein kinase-activated protein kinase-2 (MK2) is known to play a pivotal role in inflammatory processes. Here, we studied the effect of MK2 deficiency and tumor necrosis factor (TNF)-alpha-deficiency on skin tumor development in mice using the two-stage chemical carcinogenesis model. We found that MK2(-/-) mice developed significantly fewer skin tumors compared with both TNF-alpha(-/-) and wild-type mice when induced by initiation with 7,12 dimethylbenz[a]anthracene (DMBA) and by promotion with 12-O-tetradecanoylphorbol 13-acetate (TPA). The TPA-induced inflammatory response was reduced in both, TNF alpha(-/-) mice and MK2(-/-) mice, but most pronounced in TNF-alpha(-/-) mice, indicating that a reduced inflammatory response was not the only explanation for the inhibited tumorigenesis seen in MK2(-/-) mice. Interestingly, increased numbers of apoptotic cells were detected in the epidermis of MK2(-/-) mice compared with TNF-alpha(-/-) and wild-type mice, suggesting an additional role of MK2 in the regulation of apoptosis. This was further supported by: (i) increased levels of the tumor suppressor protein p53 in MK2(-/-) mice after DMBA/TPA treatment compared with controls, (ii) reduced phosphorylation (activation) of the negative p53 regulator, murine double minute 2 in MK2(-)(/-) mouse keratinocytes in vitro and (iii) a significant decrease in the DMBA/TPA induced apoptosis in cultured MK2(-/-) keratinocytes transfected with p53 small interfering RNA. Taken together, these findings demonstrate a dual role of MK2 in the early stages of tumor promotion through regulation of both the inflammatory response and apoptosis of DNA-damaged cells. These results also identify MK2 as a putative target for future skin carcinoma therapy. PMID- 19808858 TI - Adrenocortical tumor with two distinct elements revealed by combined (18)F fluorodeoxyglucose positron emission tomography and (131)I nor-cholesterol scintigraphy. PMID- 19808859 TI - Early Origins of polycystic ovary syndrome: hypotheses may change without notice. PMID- 19808860 TI - Who's assessing tobacco use in cancer clinical trials? AB - INTRODUCTION: Clinical trials that do not collect data on tobacco use/exposure may not adequately assess the efficacy and effectiveness of experimental treatments. METHODS: A cross-sectional study of interventional trials cited on ClinicalTrials.gov was undertaken that inquired of Local Project Directors from Connecticut guiding studies of breast, prostate, or colorectal cancer chemotherapy (N = 68) whether their protocols measured tobacco use by trial participants. Information pertaining to 46 trials (68%) is reported here. All but 1 were multicentered trials enrolling patients around the country. RESULTS: Only 3 trials (7%) reported routine collection of tobacco use information at baseline and no trial reported monitoring tobacco use during treatment follow-up. None of the 3 trials collecting tobacco data reported using exposure information in analysis of treatment effects. Survey respondents suggested that uncertainty about the relevance of tobacco exposure to therapeutic efficacy, ambivalence about how to incorporate such data into analyses, insufficient resources for collecting such information, and uncertainty about the validity of assessment methods might be reasons why tobacco use is not routinely assessed. DISCUSSION: Additional studies that address a fuller range of cancers, therapies, disease states, and clinical environments are needed to fully define the extent of this data lapse. Providing clinicians and trialists with appropriate tools for tobacco use assessment and encouraging them to collect such information about patients during treatment and follow-up may offer a simple cost-effective way to improve the quality and consequences of cancer care for every patient. PMID- 19808861 TI - A method comparison study of timeline followback and ecological momentary assessment of daily cigarette consumption. AB - INTRODUCTION: Uncertainty exists about how best to measure daily cigarette consumption. Two common measures are timeline followback (TLFB), which involves structured, prompted recall, and ecological momentary assessment (EMA), which involves recording consumption, as it occurs, on a handheld electronic device. METHODS: We evaluated the agreement between TLFB and EMA measures collected for 14 days prior to the target quit date from 236 smokers in a smoking cessation program. We performed a Bland-Altman analysis to assess agreement of TLFB and EMA using a regression-based model that allows for a nonuniform difference between methods and limits of agreement that can vary with the number of cigarettes smoked. RESULTS: For pairs of measurements taken on the same smoker, TLFB counts were on average 3.2 cigarettes higher than EMA counts; this difference increased for larger numbers of cigarettes. Using a model that allows for variable limits of agreement, the width of the 95% interval ranged from 8.7 to 61.8 cigarettes, with an average of 26.4 cigarettes. Variation between the methods increased substantially for larger cigarette counts, leading to wider limits and poorer agreement for heavy smokers. DISCUSSION: Throughout the measurement range, the estimated limits of agreement were far wider than the limits of clinical significance, defined a priori to be 20% of the number of cigarettes smoked. We conclude that TLFB and EMA cannot be considered equivalent for the assessment of daily cigarette consumption, especially for heavy smokers. PMID- 19808862 TI - Use of gene expression of neural markers in cultured neural cells to identify developmental neurotoxicants. PMID- 19808863 TI - P19 neuronal differentiation and retinoic acid metabolism as criteria to investigate atrazine, nitrite, and nitrate developmental toxicity. AB - Atrazine and nitrogenous fertilizers are agrochemical contaminants frequently detected in water systems in North America. Several studies reported their ability to affect amphibian and mammalian development. Retinoids, supplied in the diet or synthesized by cells, are essential to embryogenesis. Disturbance of their homeostasis may lead to teratogenic effects. Retinoic acid (RA) is a major retinoid regulator of cell proliferation and differentiation. Previous studies reported alterations of retinoid stores in bullfrogs of Yamaska River subwatersheds (Quebec, Canada), a region of intensive agricultural activities associated with atrazine, nitrate, and nitrite contaminants. These contaminants could affect RA metabolism and RA-mediated processes. Mouse P19 embryonic stem cells, which can differentiate to neurons in response to RA, were used to test this hypothesis. Cells were cultured in the absence or presence of contaminants during neuroinduction with RA and assayed by flow cytometry for expression of stage-specific embryonic antigen-1 (SSEA1) (embryonic marker) and betaIII-tubulin (neuronal marker). Cell cultures were also analyzed for RA metabolism by high performance liquid chromotagraphy (HPLC). Downregulation of SSEA1 paralleled betaIII-tubulin upregulation in an RA concentration-dependent manner. Atrazine, nitrate, and nitrite did not affect differentiation at environmentally encountered micromolar concentrations. However, low molar nitrite prevented RA induced SSEA1 downregulation and decreased betaIII-tubulin appearance. Decreased cell viability/proliferation accompanied these differentiation effects. P19 cells metabolized RA to polar retinoids. RA metabolism was not affected at any concentration of atrazine, nitrate, or nitrite. Environmentally relevant levels of these contaminants, thus, had no gross effect on neurodifferentiation and RA catabolism of embryonic stem cells. P19 cell-based bioassays may provide valuable tools in monitoring developmental toxicity. PMID- 19808864 TI - Infrequent transitions between saline and fresh waters in one of the most abundant microbial lineages (SAR11). AB - The aquatic bacterial group SAR11 is one of the most abundant organisms on Earth, with an estimated global population size of 2.4 x 10(28) cells in the oceans. Members of SAR11 have also been detected in brackish and fresh waters, but the evolutionary relationships between the species present in the different environments have been ambiguous. In particular, it was not clear how frequently this lineage has crossed the saline-freshwater boundary during its evolutionary diversification. Due to the huge population size of SAR11 and the potential of microbes for long-distance dispersal, we hypothesized that environmental transitions could have occurred repeatedly during the evolutionary diversification of this group. Here, we have constructed extensive 16S rDNA-based molecular phylogenies and undertaken metagenomic data analyses to assess the frequency of saline-freshwater transitions in SAR11 and to investigate the evolutionary implications of this process. Our analyses indicated that very few saline-freshwater transitions occurred during the evolutionary diversification of SAR11, generating genetically distinct saline and freshwater lineages that do not appear to exchange genes extensively via horizontal gene transfer. In contrast to lineages from saline environments, extant freshwater taxa from diverse, and sometimes distant, geographic locations were very closely related. This points to a rapid diversification and dispersal in fresh waters or to slower evolutionary rates in fresh water SAR11 when compared with marine counterparts. In addition, the colonization of both saline and fresh waters appears to have occurred early in the evolution of SAR11. We conclude that the different biogeochemical conditions that prevail in saline and fresh waters have likely prevented the environmental transitions in SAR11, promoting the evolution of clearly distinct lineages in each environment. PMID- 19808865 TI - Inference and characterization of horizontally transferred gene families using stochastic mapping. AB - Macrogenomic events, in which genes are gained and lost, play a pivotal evolutionary role in microbial evolution. Nevertheless, probabilistic evolutionary models describing such events and methods for their robust inference are considerably less developed than existing methodologies for analyzing site specific sequence evolution. Here, we present a novel method for the inference of gains and losses of gene families. First, we develop probabilistic-evolutionary models describing the dynamics of gene-family content, which are more biologically realistic than previously suggested models. In our likelihood-based models, gains and losses are represented by transitions between presence and absence, given an underlying phylogeny. We employ a mixture-model approach in which we allow both the gain rate and the loss rate to vary among gene families. Second, we use these models together with the analytic implementation of stochastic mapping to infer branch-specific events. Our novel methodology allows us to infer and quantify horizontal gene transfer (HGT) events. This enables us to rank various gene families and lineages according to their propensity to undergo gains and losses. Applying our methodology to 4,873 gene families shows that: 1) the novel mixture models describe the observed variability in gene family content among microbes significantly better than previous models; 2) The stochastic mapping approach enables accurate inference of gain and loss events based on simulations; 3) At least 34% of the gene families analyzed are inferred to have experienced HGT at least once during their evolution; and 4) Gene families that were inferred to experience HGT are both enriched and depleted with respect to specific functional categories. PMID- 19808866 TI - Oral poly(ADP-ribose) polymerase-1 inhibitor BSI-401 has antitumor activity and synergizes with oxaliplatin against pancreatic cancer, preventing acute neurotoxicity. AB - PURPOSE: Development of novel agents and drug combinations are urgently needed for treatment of pancreatic cancer. Oxaliplatin belongs to an important class of DNA-damaging organoplatinum agents, useful in pancreatic cancer therapy. However, increased ability of cancer cells to recognize and repair DNA damage enables resistance to these agents. Poly (ADP ribose) polymerase-1 is a sensor of DNA damage with key roles in DNA repair. Here, we report the therapeutic activity of the poly (ADP ribose) polymerase-1 inhibitor BSI-401, as a single agent and in combination with oxaliplatin in orthotopic nude mouse models of pancreatic cancer, and its effect on oxaliplatin-induced acute neurotoxicity. EXPERIMENTAL DESIGN: We determined in vitro the effect of BSI-401 and its synergism with oxaliplatin on the growth of pancreatic cancer cells. Activity of different dosages of parenteral and oral BSI-401, alone and in combination with oxaliplatin, was evaluated in orthotopic nude mouse models with luciferase expressing pancreatic cancer cells. The effect of BSI-401 in preventing oxaliplatin-induced acute cold allodynia was measured in rats using a temperature controlled plate. RESULTS: BSI-401 alone and in synergism with oxaliplatin significantly inhibited the growth of pancreatic cancer cells in vitro. In nude mice, i.p. [200 mg/kg once a week (QW) x 4] and oral [400 mg/kg days 1-5 of each week (QD5 + R2) x 4] administration of BSI-401 significantly reduced tumor burden and prolonged survival (46 versus 144 days, P = 0.0018; 73 versus 194 days, P = 0.0017) compared with no treatment. BSI-401 combined with oxaliplatin had potent synergistic antitumor activity (46 versus 132 days, P = 0.0063), and significantly (P = 0.0148) prevented acute oxaliplatin-induced neurotoxicity. CONCLUSIONS: BSI-401, alone or in combination with oxaliplatin, is a promising new therapeutic agent that warrants further evaluation for treatment of pancreatic cancer. PMID- 19808867 TI - Polysomy for chromosomes 1 and 19 predicts earlier recurrence in anaplastic oligodendrogliomas with concurrent 1p/19q loss. AB - PURPOSE: Loss of chromosome arms 1p and 19q is a molecular feature of oligodendroglial tumors characterized by responsiveness to chemotherapy and a favorable prognosis. The purpose of this study was to evaluate the prognostic significance of polysomy of chromosomes 1 and 19 in the setting of 1p/19q codeletion. EXPERIMENTAL DESIGN: We analyzed 64 anaplastic oligodendrogliomas with 1p/19q loss or maintenance diagnosed at Massachusetts General Hospital and Brigham and Women's Hospital from 1996 to 2005; fluorescence in situ hybridization for 1p/19q and Ki-67 immunohistochemistry was done. Polysomy was defined as more than two 1q and 19p signals in >30% of the cells with concurrent 1p/19q deletion. Tumors were divided into groups based on their 1p/19q status and compared for progression-free survival, overall survival, and 5-year survival probabilities. RESULTS: Forty-six tumors (72%) in our cohort had 1p/19q loss and 18 (28%) had 1p/19q maintenance. Of those with loss, 19 (41%) had concurrent polysomy and 27 (59%) lacked polysomy. In agreement with previous studies, the group of anaplastic oligodendrogliomas with 1p/19q loss had significantly better progression-free survival and overall survival than anaplastic oligodendrogliomas with 1p/19q maintenance (P = 0.0009 and P < 0.0003, respectively). Among anaplastic oligodendrogliomas with 1p/19q loss, those with polysomy showed shorter progression-free survival than those with 1p/19q loss without polysomy (P = 0.0048). Overall survival was similar in tumors with and without polysomy. The Ki-67 labeling index was not associated with polysomy and did not have prognostic significance. CONCLUSION: The presence of polysomy in anaplastic oligodendrogliomas with deletion of 1p/19q is a marker of earlier recurrence. PMID- 19808868 TI - Therapy-induced PML/RARA proteolysis and acute promyelocytic leukemia cure. AB - Acute promyelocytic leukemia (APL) is characterized by a specific t(15;17) chromosomal translocation that yields the PML/RARA fusion gene. Clinically, besides chemotherapy, two drugs induce clinical remissions: retinoic acid (RA) and arsenic trioxide (As). Both agents directly target PML/RARA-mediated transcriptional repression and protein stability, inducing to various extent promyelocyte differentiation and clinical remission of APL patients. RA targets the RARA moiety of the fusion, whereas arsenic targets its PML part. PML/RARA expression in the mouse is sufficient to initiate APL. The RA-As association, which synergizes for PML/RARA degradation but not for differentiation, rapidly clears leukemia initiating cells (LIC), resulting in APL eradication in murine APL models, but also in several APL clinical trials. Cyclic AMP triggered PML/RARA phosphorylation also enhances RA-induced APL regression, PML/RARA degradation, and LIC clearance, raising new options for therapy-resistant patients. Although differentiation has a major role in debulking of the tumor, PML/RARA degradation seems to be the primary basis for APL eradication by the RA As association. Oncoprotein degradation could be a general therapeutic strategy that may be extended beyond APL. PMID- 19808869 TI - Recent advances in cancer therapy targeting proteins involved in DNA double strand break repair. AB - Damage to genetic material represents a persistent and ubiquitous threat to genomic stability. Once DNA damage is detected, a multifaceted signaling network is activated that halts the cell cycle, initiates repair, and in some instances induces apoptotic cell death. In this article, we will review DNA damage surveillance networks, which maintain the stability of our genome, and discuss the efforts underway to identify chemotherapeutic compounds targeting the core components of DNA double-strand breaks (DSB) response pathway. The majority of tumor cells have defects in maintaining genomic stability owing to the loss of an appropriate response to DNA damage. New anticancer agents are exploiting this vulnerability of cancer cells to enhance therapeutic indexes, with limited normal tissue toxicity. Recently inhibitors of the checkpoint kinases Chk1 and Chk2 have been shown to sensitize tumor cells to DNA damaging agents. In addition, the treatment of BRCA1- or BRCA2-deficient tumor cells with poly(ADP-ribose) polymerase (PARP) inhibitors also leads to specific tumor killing. Due to the numerous roles of p53 in genomic stability and its defects in many human cancers, therapeutic agents that restore p53 activity in tumors are the subject of multiple clinical trials. In this article we highlight the proteins mentioned above and catalog several additional players in the DNA damage response pathway, including ATM, DNA-PK, and the MRN complex, which might be amenable to pharmacological interventions and lead to new approaches to sensitize cancer cells to radio- and chemotherapy. The challenge is how to identify those patients most receptive to these treatments. PMID- 19808870 TI - Identification of novel kinase targets for the treatment of estrogen receptor negative breast cancer. AB - PURPOSE: Previous gene expression profiling studies of breast cancer have focused on the entire genome to identify genes differentially expressed between estrogen receptor (ER) alpha-positive and ER-alpha-negative cancers. EXPERIMENTAL DESIGN: Here, we used gene expression microarray profiling to identify a distinct kinase gene expression profile that identifies ER-negative breast tumors and subsets ER negative breast tumors into four distinct subtypes. RESULTS: Based on the types of kinases expressed in these clusters, we identify a cell cycle regulatory subset, a S6 kinase pathway cluster, an immunomodulatory kinase-expressing cluster, and a mitogen-activated protein kinase pathway cluster. Furthermore, we show that this specific kinase profile is validated using independent sets of human tumors and is also seen in a panel of breast cancer cell lines. Kinase expression knockdown studies show that many of these kinases are essential for the growth of ER-negative, but not ER-positive, breast cancer cell lines. Finally, survival analysis of patients with breast cancer shows that the S6 kinase pathway signature subtype of ER-negative cancers confers an extremely poor prognosis, whereas patients whose tumors express high levels of immunomodulatory kinases have a significantly better prognosis. CONCLUSIONS: This study identifies a list of kinases that are prognostic and may serve as druggable targets for the treatment of ER-negative breast cancer. PMID- 19808871 TI - An embryonic stem cell-like signature identifies poorly differentiated lung adenocarcinoma but not squamous cell carcinoma. AB - PURPOSE: An embryonic stem cell (ESC) profile correlates with poorly differentiated breast, bladder, and glioma cancers. In this article, we assess the correlation between the ESC profile and clinical variables in lung cancer. EXPERIMENTAL DESIGN: Microarray gene expression analysis was done using Affymetrix Human Genome U133A on 443 samples of human lung adenocarcinoma and 130 samples of squamous cell carcinoma (SCC). To identify gene set enrichment patterns, we used the Genomica software. RESULTS: Our analysis showed that an increased expression of the ESC gene set and a decreased expression of the Polycomb target gene set identified poorly differentiated lung adenocarcinoma. In addition, this gene expression signature was associated with markers of poor prognosis and worse overall survival in lung adenocarcinoma. However, there was no correlation between this ESC gene signature and any histologic or clinical variable assessed in lung SCC. CONCLUSIONS: This work suggests that not all poorly differentiated non-small cell lung cancers exhibit a gene expression profile similar to that of ESC, and that other characteristics may play a more important role in the determination of differentiation and survival in SCC of the lung. PMID- 19808872 TI - Loss of SNF5 expression correlates with poor patient survival in melanoma. AB - PURPOSE: Aberrant expression of SWI/SNF chromatin remodeling complex is involved in cancer development. The tumor suppressor SNF5, the core subunit of SWI/SNF complex, has been shown to regulate cell differentiation, cell cycle control, and apoptosis. To investigate the role of SNF5 in the development of melanoma, we examined the expression of SNF5 in melanocytic lesions at different stages and analyzed the correlation between SNF5 expression and clinicopathologic variables and patient survival. EXPERIMENTAL DESIGN: Using tissue microarray and immunohistochemistry, we evaluated SNF5 staining in 51 dysplastic nevi, 88 primary melanomas, and 48 metastatic melanomas. We studied chemosensitivity of melanoma cells with reduced SNF5 expression by siRNA using cell survival and apoptosis assays. RESULTS: SNF5 expression was reduced in metastatic melanoma compared with dysplastic nevi (P = 0.005), in advanced primary melanoma (Clark's level V) compared with low risk Clark's level II melanoma (P = 0.019), and in melanoma at sun-exposed sites compared with sun-protected sites (P = 0.044). Furthermore, we showed a strong correlation between negative SNF5 expression and a worse 5-year survival in melanoma patients (P = 0.016). Multivariate Cox regression analysis revealed that negative SNF5 expression is an independent prognostic factor to predict patient outcome in primary melanomas (P = 0.031). Finally, we showed that knockdown of SNF5 in melanoma cell lines resulted in significant chemoresistance. CONCLUSIONS: Our data indicate that SNF5 may be an important marker for human melanoma progression and prognosis as well as a potential therapeutic target. PMID- 19808873 TI - Dose-ranging study of metronomic oral vinorelbine in patients with advanced refractory cancer. AB - AIM: To determine the safe dose range and pharmacokinetics of metronomic oral vinorelbine and obtain preliminary data on biomarkers and efficacy in patients with advanced cancer. METHODS: Successive cohorts of patients received escalated doses of oral vinorelbine given thrice a week until disease progression, unacceptable toxicity (UT), or consent withdrawal. UT was any grade 4 toxicity, or grade 2 or 3 toxicity that would result to longer than 2-week break during the first 2 months of treatment. Blood samples were collected for pharmacokinetics and quantification of angiogenesis regulatory proteins. RESULTS: Sixty-two patients (median age, 60 years) enrolled at six dose levels from 20 to 70 mg and received treatment for median 12.25 weeks (range, 2-216+). Unacceptable toxicity occurred in two of six patients treated at 60 mg (leucopenia grade 4 and epistaxis grade 2) and in one at 70 mg (leucopenia grade 2). The upper metronomic dose was 50 mg. Objective antitumor response documented in eight cases and 32% of patients experienced disease stability for minimum 6 months. Three responders (renal cancer, medullary thyroid carcinoma, and Kaposi sarcoma) received nonstop treatment for over 3 years without overt toxicity. Low pretreatment levels of circulating interleukin-8, vascular endothelial growth factor, and basic fibroblast growth factor were found predictors of efficacy. Steady-state concentrations of vinorelbine and its active metabolite ranged from 0.5 to 1.5 ng/mL. CONCLUSIONS: Metronomic administration of oral vinorelbine is feasible at doses up to 50 mg thrice a week and can yield sustainable antitumor activity without overt toxicity, probably through antiangiogenic mechanism. Further clinical investigation is warranted. PMID- 19808874 TI - Phase I study of ipilimumab, an anti-CTLA-4 monoclonal antibody, in patients with relapsed and refractory B-cell non-Hodgkin lymphoma. AB - PURPOSE: The growth of non-Hodgkin lymphomas can be influenced by tumor-immune system interactions. Cytotoxic T-lymphocyte antigen 4 (CTLA-4) is a negative regulator of T-cell activation that serves to dampen antitumor immune responses. Blocking anti-CTLA-4 monoclonal antibodies improves host resistance to immunogenic tumors, and the anti-CTLA-4 antibody ipilimumab (MDX-010) has clinical activity against melanoma, prostate, and ovarian cancers. EXPERIMENTAL DESIGN: We did a phase I trial of ipilimumab in patients with relapsed/refractory B-cell lymphoma to evaluate safety, immunologic activity, and potential clinical efficacy. Treatment consisted of ipilimumab at 3 mg/kg and then monthly at 1 mg/kg x 3 months (dose level 1), with subsequent escalation to 3 mg/kg monthly x 4 months (dose level 2). RESULTS: Eighteen patients were treated, 12 at the lower dose level and 6 at the higher dose level. Ipilimumab was generally well tolerated, with common adverse events attributed to it, including diarrhea, headache, abdominal pain, anorexia, fatigue, neutropenia, and thrombocytopenia. Two patients had clinical responses; one patient with diffuse large B-cell lymphoma had an ongoing complete response (>31 months), and one with follicular lymphoma had a partial response lasting 19 months. In 5 of 16 cases tested (31%), T-cell proliferation to recall antigens was significantly increased (>2-fold) after ipilimumab therapy. CONCLUSIONS: Blockade of CTLA-4 signaling with the use of ipilimumab is well tolerated at the doses used and has antitumor activity in patients with B-cell lymphoma. Further evaluation of ipilimumab alone or in combination with other agents in B-cell lymphoma patients is therefore warranted. PMID- 19808875 TI - Qupe--a Rich Internet Application to take a step forward in the analysis of mass spectrometry-based quantitative proteomics experiments. AB - MOTIVATION: The goal of present -omics sciences is to understand biological systems as a whole in terms of interactions of the individual cellular components. One of the main building blocks in this field of study is proteomics where tandem mass spectrometry (LC-MS/MS) in combination with isotopic labelling techniques provides a common way to obtain a direct insight into regulation at the protein level. Methods to identify and quantify the peptides contained in a sample are well established, and their output usually results in lists of identified proteins and calculated relative abundance values. The next step is to move ahead from these abstract lists and apply statistical inference methods to compare measurements, to identify genes that are significantly up- or down regulated, or to detect clusters of proteins with similar expression profiles. RESULTS: We introduce the Rich Internet Application (RIA) Qupe providing comprehensive data management and analysis functions for LC-MS/MS experiments. Starting with the import of mass spectra data the system guides the experimenter through the process of protein identification by database search, the calculation of protein abundance ratios, and in particular, the statistical evaluation of the quantification results including multivariate analysis methods such as analysis of variance or hierarchical cluster analysis. While a data model to store these results has been developed, a well-defined programming interface facilitates the integration of novel approaches. A compute cluster is utilized to distribute computationally intensive calculations, and a web service allows to interchange information with other -omics software applications. To demonstrate that Qupe represents a step forward in quantitative proteomics analysis an application study on Corynebacterium glutamicum has been carried out. AVAILABILITY AND IMPLEMENTATION: Qupe is implemented in Java utilizing Hibernate, Echo2, R and the Spring framework. We encourage the usage of the RIA in the sense of the 'software as a service' concept, maintained on our servers and accessible at the following location: http://qupe.cebitec.uni-bielefeld.de. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19808876 TI - CentroidAlign: fast and accurate aligner for structured RNAs by maximizing expected sum-of-pairs score. AB - MOTIVATION: The importance of accurate and fast predictions of multiple alignments for RNA sequences has increased due to recent findings about functional non-coding RNAs. Recent studies suggest that maximizing the expected accuracy of predictions will be useful for many problems in bioinformatics. RESULTS: We designed a novel estimator for multiple alignments of structured RNAs, based on maximizing the expected accuracy of predictions. First, we define the maximum expected accuracy (MEA) estimator for pairwise alignment of RNA sequences. This maximizes the expected sum-of-pairs score (SPS) of a predicted alignment under a probability distribution of alignments given by marginalizing the Sankoff model. Then, by approximating the MEA estimator, we obtain an estimator whose time complexity is O(L(3)+c(2)dL(2)) where L is the length of input sequences and both c and d are constants independent of L. The proposed estimator can handle uncertainty of secondary structures and alignments that are obstacles in Bioinformatics because it considers all the secondary structures and all the pairwise alignments as input sequences. Moreover, we integrate the probabilistic consistency transformation (PCT) on alignments into the proposed estimator. Computational experiments using six benchmark datasets indicate that the proposed method achieved a favorable SPS and was the fastest of many state-of the-art tools for multiple alignments of structured RNAs. AVAILABILITY: The software called CentroidAlign, which is an implementation of the algorithm in this article, is freely available on our website: http://www.ncrna.org/software/centroidalign/. CONTACT: hamada-michiaki@aist.go.jp SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19808877 TI - Effect of read-mapping biases on detecting allele-specific expression from RNA sequencing data. AB - MOTIVATION: Next-generation sequencing has become an important tool for genome wide quantification of DNA and RNA. However, a major technical hurdle lies in the need to map short sequence reads back to their correct locations in a reference genome. Here, we investigate the impact of SNP variation on the reliability of read-mapping in the context of detecting allele-specific expression (ASE). RESULTS: We generated 16 million 35 bp reads from mRNA of each of two HapMap Yoruba individuals. When we mapped these reads to the human genome we found that, at heterozygous SNPs, there was a significant bias toward higher mapping rates of the allele in the reference sequence, compared with the alternative allele. Masking known SNP positions in the genome sequence eliminated the reference bias but, surprisingly, did not lead to more reliable results overall. We find that even after masking, approximately 5-10% of SNPs still have an inherent bias toward more effective mapping of one allele. Filtering out inherently biased SNPs removes 40% of the top signals of ASE. The remaining SNPs showing ASE are enriched in genes previously known to harbor cis-regulatory variation or known to show uniparental imprinting. Our results have implications for a variety of applications involving detection of alternate alleles from short-read sequence data. AVAILABILITY: Scripts, written in Perl and R, for simulating short reads, masking SNP variation in a reference genome and analyzing the simulation output are available upon request from JFD. Raw short read data were deposited in GEO (http://www.ncbi.nlm.nih.gov/geo/) under accession number GSE18156. CONTACT: jdegner@uchicago.edu; marioni@uchicago.edu; gilad@uchicago.edu; pritch@uchicago.edu SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19808878 TI - Quantitative measurement of aging using image texture entropy. AB - MOTIVATION: A key element in understanding the aging of Caenorhabditis elegans is objective quantification of the morphological differences between younger and older animals. Here we propose to use the image texture entropy as an objective measurement that reflects the structural deterioration of the C. elegans muscle tissues during aging. RESULTS: The texture entropy and directionality of the muscle microscopy images were measured using 50 animals on Days 0, 2, 4, 6, 8, 10 and 12 of adulthood. Results show that the entropy of the C. elegans pharynx tissues increases as the animal ages, but a sharper increase was measured between Days 2 and 4, and between Days 8 and 10. These results are in agreement with gene expression findings, and support the contention that the process of C. elegans aging has several distinct stages. This can indicate that C. elegans aging is driven by developmental pathways, rather than stochastic accumulation of damage. AVAILABILITY: The image data are freely available on the Internet at http://ome.grc.nia.nih.gov/iicbu2008/celegans, and the Haralick and Tamura texture analysis source code can be downloaded at http://ome.grc.nia.nih.gov/wnd charm. PMID- 19808879 TI - Exon Array Analyzer: a web interface for Affymetrix exon array analysis. AB - SUMMARY: The Exon Array Analyzer (EAA) is a web server, which provides a user friendly interface to identify alternative splicing events analyzed with Affymetrix Exon Arrays. The EAA implements the Splice Index algorithm to identify differential expressed exons. The use of various filters allows reduction of the number of false positive hits. Results are presented with detailed annotation information and graphics to identify splice events and to facilitate biological validations. To demonstrate the versatility of the EAA, we analyzed exon arrays of 11 different murine tissues using sample data provided by Affymetrix (http://www.affymetrix.com). Data from the heart were compared with other tissues to identify exons that undergo heart-specific alternatively splicing, resulting in the identification of 885 differentially expressed probe sets in 649 genes. AVAILABILITY: The web interface is available at http://EAA.mpi-bn.mpg.de/. Detailed documentation is available on the EAA web site (http://EAA.mpi bn.mpg.de/supp.php) including screen shots, example analyzes and step by step instructions. CONTACT: thomas.braun@mpi-bn.mpg.de SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19808880 TI - HTqPCR: high-throughput analysis and visualization of quantitative real-time PCR data in R. AB - MOTIVATION: Quantitative real-time polymerase chain reaction (qPCR) is routinely used for RNA expression profiling, validation of microarray hybridization data and clinical diagnostic assays. Although numerous statistical tools are available in the public domain for the analysis of microarray experiments, this is not the case for qPCR. Proprietary software is typically provided by instrument manufacturers, but these solutions are not amenable to the tandem analysis of multiple assays. This is problematic when an experiment involves more than a simple comparison between a control and treatment sample, or when many qPCR datasets are to be analyzed in a high-throughput facility. RESULTS: We have developed HTqPCR, a package for the R statistical computing environment, to enable the processing and analysis of qPCR data across multiple conditions and replicates. AVAILABILITY: HTqPCR and user documentation can be obtained through Bioconductor or at http://www.ebi.ac.uk/bertone/software. CONTACT: bertone@ebi.ac.uk PMID- 19808881 TI - Pathway identification by network pruning in the metabolic network of Escherichia coli. AB - MOTIVATION: All metabolic networks contain metabolites, such as ATP and NAD, known as currency metabolites, which take part in many reactions. These are often removed in the study of these networks, but no consensus exists on what actually constitutes a currency metabolite, and it is also unclear how these highly connected nodes contribute to the global structure of the network. RESULTS: In this article, we analyse how the Escherichia coli metabolic network responds to pruning in the form of sequential removal of metabolites with highest degree. As expected this leads to network fragmentation, but the process by which it occurs suggests modularity and long-range correlations within the network. We find that the pruned networks contain longer paths than the random expectation, and that the paths that survive the pruning also exhibit a lower cost (number of involved metabolites) compared with random paths in the full metabolic network. Finally we confirm that paths detected by pruning overlap with known metabolic pathways. We conclude that pruning reveals functional pathways in metabolic networks, where currency metabolites may be seen as ingredients in a well-balanced soup in which main metabolic production lines are immersed. CONTACT: gerlee@nbi.dk SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19808882 TI - Adaptive multi-agent architecture for functional sequence motifs recognition. AB - MOTIVATION: Accurate genome annotation or protein function prediction requires precise recognition of functional sequence motifs. Many computational motif prediction models have been proposed. Due to the complexity of the biological data, it may be desirable to apply an integrated approach that uses multiple models for analysis. RESULTS: In this article, we propose a novel multi-agent architecture for the general purpose of functional sequence motif recognition. The approach takes advantage of the synergy provided by multiple agents through the employment of different agents equipped with distinctive problem solving skills and promotes the collaborations among them through decision maker (DM) agents that work as classifier ensembles. A genetic algorithm-based fusion strategy is applied which offers evolutionary property to the DM agents. The consistency and robustness of the system are maintained by an evolvable agent that mediates the team of the ensemble agents. The combined effort of a recommendation system (Seer) and the self-learning mediator agent yields a successful identification of the most efficient agent deployment scheme at an early stage of the experimentation process, which has the potential of greatly reducing the computational cost of the system. Two concrete systems are constructed that aim at predicting two important sequence motifs-the translational initiation sites (TISs) and the core promoters. With the incorporation of three distinctive problem solver agents, the TIS predictor consistently outperforms most of the state-of-the-art approaches under investigation. Integrating three existing promoter predictors, our system is able to yield consistently good performance. AVAILABILITY: The program (MotifMAS) and the datasets are available upon request. PMID- 19808883 TI - ncRNAppi--a tool for identifying disease-related miRNA and siRNA targeting pathways. AB - Currently, there are a number of databases which store microRNA (miRNA) information, and tools available which provide miRNA target prediction. In this article, we describe a novel web-based tool that integrate the miRNA-targeted mRNA data, protein-protein interactions (PPI) records, tissues, biochemical pathways, human disease and gene function information to establish a disease related miRNA target pathway database. This database is unique in the sense that it links miRNA target genes with their PPI partners according to being tissue- and diseases-specific or both. The same approach is also applied to siRNA data. This database provides two types of searches: (i) tissue- and (ii) disease specific miRNA (or siRNA) targeting pathways. The search allows one to identify tissue- or disease-specific miRNA (or siRNA) target gene's PPI partners two levels beyond. AVAILABILITY: The release version 1.0 is a freely accessible database available at http://ncrnappi.cs.nthu.edu.tw and http://ncRNAppi.bioinfo.asia.edu.tw/. PMID- 19808884 TI - Mouse oviduct-specific glycoprotein is an egg-associated ZP3-independent sperm adhesion ligand. AB - Mouse sperm-egg binding requires a multiplicity of receptor-ligand interactions, including an oviduct-derived, high molecular weight, wheat germ agglutinin (WGA) binding glycoprotein that associates with the egg coat at ovulation. Herein, we report the purification and identification of this sperm-binding ligand. WGA binding, high molecular weight glycoproteins isolated from hormonally primed mouse oviduct lysates competitively inhibit sperm-egg binding in vitro. Within this heterogeneous glycoprotein preparation, a distinct 220 kDa protein selectively binds to sperm surfaces, and was identified by sequence analysis as oviduct-specific glycoprotein (OGP). The sperm-binding activity of OGP was confirmed by the loss of sperm-binding following immunodepletion of OGP from oviduct lysates, and by the ability of both immunoprecipitated OGP and natively purified OGP to competitively inhibit sperm-egg binding. As expected, OGP is expressed by the secretory cells of the fimbriae and infundibulum; however, in contrast to previous reports, OGP is also associated with both the zona pellucida and the perivitelline space of mouse oocytes. Western blot analysis and lectin affinity chromatography demonstrate that whereas the bulk of OGP remains soluble in the ampullar fluid, distinct glycoforms associate with the cumulus matrix, zona pellucida and perivitelline space. The sperm-binding activity of OGP is carbohydrate-dependent and restricted to a relatively minor peanut agglutinin (PNA)-binding glycoform that preferentially associates with the sperm surface, zona pellucida and perivitelline space, relative to other more abundant glycoforms. Finally, pretreatment of two-cell embryos, which do not normally bind sperm, with PNA-binding OGP stimulates sperm binding. PMID- 19808885 TI - Nuclear localisation is crucial for the proapoptotic activity of the HtrA-like serine protease Nma111p. AB - Programmed cell death is induced by the activation of a subset of intracellular proteins in response to specific extra- and intracellular signals. In the yeast Saccharomyces cerevisiae, Nma111p functions as a nuclear serine protease that is necessary for apoptosis under cellular stress conditions, such as elevated temperature or treatment of cells with hydrogen peroxide to induce cell death. We have examined the role of nuclear protein import in the function of Nma111p in apoptosis. Nma111p contains two small clusters of basic residues towards its N terminus, both of which are necessary for efficient translocation into the nucleus. Nma111p does not shuttle between the nucleus and cytoplasm during either normal growth conditions or under environmental stresses that induce apoptosis. The N-terminal half of Nma111p is sufficient to provide the apoptosis-inducing activity of the protein, and the nuclear-localisation signal (NLS) sequences and catalytic serine 235 are both necessary for this function. We provide compelling evidence that intranuclear Nma111p activity is necessary for apoptosis in yeast. PMID- 19808886 TI - Myosin V spatially regulates microtubule dynamics and promotes the ubiquitin dependent degradation of the fission yeast CLIP-170 homologue, Tip1. AB - Coordination between microtubule and actin cytoskeletons plays a crucial role during the establishment of cell polarity. In fission yeast, the microtubule cytoskeleton regulates the distribution of actin assembly at the new growing end during the monopolar-to-bipolar growth transition. Here, we describe a novel mechanism in which a myosin V modulates the spatial coordination of proteolysis and microtubule dynamics. In cells lacking a functional copy of the class V myosin, Myo52, the plus ends of microtubules fail to undergo catastrophe on contacting the cell end and continue to grow, curling around the end of the cell. We show that this actin-associated motor regulates the efficient ubiquitin dependent proteolysis of the Schizosaccharomyces pombe CLIP-170 homologue, Tip1. Myo52 facilitates microtubule catastrophe by enhancing Tip1 removal from the plus end of growing microtubules at the cell tips. There, Myo52 and the ubiquitin receptor, Dph1, work in concert to target Tip1 for degradation. PMID- 19808887 TI - Ste20-kinase-dependent TEDS-site phosphorylation modulates the dynamic localisation and endocytic function of the fission yeast class I myosin, Myo1. AB - Type I myosins are monomeric motors involved in a range of motile and sensory activities in different cell types. In simple unicellular eukaryotes, motor activity of class I myosins is regulated by phosphorylation of a conserved 'TEDS site' residue within the motor domain. The mechanism by which this phosphorylation event affects the cellular function of each myosin I remains unclear. The fission yeast myosin I, Myo1, activates Arp2/3-dependent polymerisation of cortical actin patches and also regulates endocytosis. Using mutants and Myo1-specific antibodies, we show that the phosphorylation of the Myo1 TEDS site (serine 361) plays a crucial role in regulating this protein's dynamic localisation and cellular function. We conclude that although phosphorylation of serine 361 does not affect the ability of this motor protein to promote actin polymerisation, it is required for Myo1 to recruit to sites of endocytosis and function during this process. PMID- 19808888 TI - Ubiquitylation of the gap junction protein connexin-43 signals its trafficking from early endosomes to lysosomes in a process mediated by Hrs and Tsg101. AB - Gap junctions are dynamic plasma membrane domains, and their protein constituents, the connexins, have a high turnover rate in most tissue types. However, the molecular mechanisms involved in degradation of gap junctions have remained largely unknown. Here, we show that ubiquitin is strongly relocalized to connexin-43 (Cx43; also known as Gja1) gap junction plaques in response to activation of protein kinase C. Cx43 remained ubiquitylated during its transition to a Triton X-100-soluble state and along its trafficking to early endosomes. Following internalization, Cx43 partly colocalized with the ubiquitin-binding proteins Hrs (hepatocyte growth factor-regulated tyrosine kinase substrate; also known as Hgs) and Tsg101 (tumor susceptibility gene 101). Depletion of Hrs or Tsg101 by small interfering RNA abrogated trafficking of Cx43 from early endosomes to lysosomes. Under these conditions, Cx43 was able to undergo dephosphorylation and deubiquitylation, locate to the plasma membrane and form functional gap junctions. Simultaneous depletion of Hrs and Tsg101 caused accumulation of a phosphorylated and ubiquitylated subpopulation of Cx43 in early endosomes and in hybrid organelles between partly degraded annular gap junctions and endosomes. Collectively, these data reveal a central role of early endosomes in sorting of ubiquitylated Cx43, and identify Hrs and Tsg101 as crucial regulators of trafficking of Cx43 to lysosomes. PMID- 19808889 TI - Real-time measurements of cAMP production in live Dictyostelium cells. AB - Cyclic AMP has a crucial role during the entire developmental program of the social amoebae Dictyostelium, acting both as an intracellular second messenger and, when secreted, as a directional cue that is relayed to neighboring cells during chemotaxis. Although significant knowledge about cAMP production in chemotaxing cells has been derived from studies performed on cell populations, cAMP dynamics at the single cell level have not been investigated. To examine this, we used a FRET-based cAMP sensor that possesses high cAMP sensitivity and great temporal resolution. We show the transient profile of cAMP accumulation in live Dictyostelium cells and establish that chemoattractants control intracellular cAMP dynamics by regulating synthesis via the adenylyl cyclase ACA. aca(-) cells show no significant change in FRET response following chemoattractant addition. Furthermore, cells lacking ACB, the other adenylyl cyclase expressed in chemotaxing cells, behave similarly to wild-type cells. We also establish that the RegA is the major phosphodiesterase that degrades intracellular cAMP in chemotaxis-competent cells. Interestingly, we failed to measure intracellular cAMP compartmentalization in actively chemotaxing cells. We conclude that cytosolic cAMP, which is destined to activate PKA, is regulated by ACA and RegA and does not compartmentalize during chemotaxis. PMID- 19808890 TI - Regulation of podosome dynamics by WASp phosphorylation: implication in matrix degradation and chemotaxis in macrophages. AB - Podosomes, adhesion structures capable of matrix degradation, have been linked with the ability of cells to perform chemotaxis and invade tissues. Wiskott Aldrich Syndrome protein (WASp), an effector of the RhoGTPase Cdc42 and a Src family kinase substrate, regulates macrophage podosome formation. In this study, we demonstrate that WASp is active in podosomes by using TIRF-FRET microscopy. Pharmacological and RNA interference approaches suggested that continuous WASp activity is required for podosome formation and function. Rescue experiments using point mutations demonstrate an absolute requirement for Cdc42 binding to WASp in podosome formation. Although tyrosine phosphorylation was not absolutely required for podosome formation, phosphorylation did regulate the rate of podosome nucleation and actin filament stability. Importantly, WASp tyrosine phosphorylation does not alter WASp activation, instead phosphorylation appears to be important for the restriction of WASp activity to podosomes. In addition, the matrix-degrading ability of cells requires WASp phosphorylation. Chemotactic responses to CSF-1 were also attenuated in the absence of endogenous WASp, which could not be rescued with either tyrosine mutation. These results suggest a more complex role for tyrosine phosphorylation than simply in the regulation of WASp activity, and suggest a link between podosome dynamics and macrophage migration. PMID- 19808891 TI - Myosin-Va restrains the trafficking of Na+/K+-ATPase-containing vesicles in alveolar epithelial cells. AB - Stimulation of Na(+)/K(+)-ATPase activity in alveolar epithelial cells by cAMP involves its recruitment from intracellular compartments to the plasma membrane. Here, we studied the role of the actin molecular motor myosin-V in this process. We provide evidence that, in alveolar epithelial cells, cAMP promotes Na(+)/K(+) ATPase recruitment to the plasma membrane by increasing the average speed of Na(+)/K(+)-ATPase-containing vesicles moving to the cell periphery. We found that three isoforms of myosin-V are expressed in alveolar epithelial cells; however, only myosin-Va and Vc colocalized with the Na(+)/K(+)-ATPase in intracellular membrane fractions. Overexpression of dominant-negative myosin-Va or knockdown with specific shRNA increased the average speed and distance traveled by the Na(+)/K(+)-ATPase-containing vesicles, as well as the Na(+)/K(+)-ATPase activity and protein abundance at the plasma membrane to similar levels as those observed with cAMP stimulation. These data show that myosin-Va has a role in restraining Na(+)/K(+)-ATPase-containing vesicles within intracellular pools and that this restrain is released after stimulation by cAMP allowing the recruitment of the Na(+)/K(+)-ATPase to the plasma membrane and thus increased activity. PMID- 19808892 TI - Combined risk allele score of eight type 2 diabetes genes is associated with reduced first-phase glucose-stimulated insulin secretion during hyperglycemic clamps. AB - OBJECTIVE: At least 20 type 2 diabetes loci have now been identified, and several of these are associated with altered beta-cell function. In this study, we have investigated the combined effects of eight known beta-cell loci on insulin secretion stimulated by three different secretagogues during hyperglycemic clamps. RESEARCH DESIGN AND METHODS: A total of 447 subjects originating from four independent studies in the Netherlands and Germany (256 with normal glucose tolerance [NGT]/191 with impaired glucose tolerance [IGT]) underwent a hyperglycemic clamp. A subset had an extended clamp with additional glucagon-like peptide (GLP)-1 and arginine (n = 224). We next genotyped single nucleotide polymorphisms in TCF7L2, KCNJ11, CDKAL1, IGF2BP2, HHEX/IDE, CDKN2A/B, SLC30A8, and MTNR1B and calculated a risk allele score by risk allele counting. RESULTS: The risk allele score was associated with lower first-phase glucose-stimulated insulin secretion (GSIS) (P = 7.1 x 10(-6)). The effect size was equal in subjects with NGT and IGT. We also noted an inverse correlation with the disposition index (P = 1.6 x 10(-3)). When we stratified the study population according to the number of risk alleles into three groups, those with a medium- or high-risk allele score had 9 and 23% lower first-phase GSIS. Second-phase GSIS, insulin sensitivity index and GLP-1, or arginine-stimulated insulin release were not significantly different. CONCLUSIONS: A combined risk allele score for eight known beta-cell genes is associated with the rapid first-phase GSIS and the disposition index. The slower second-phase GSIS, GLP-1, and arginine-stimulated insulin secretion are not associated, suggesting that especially processes involved in rapid granule recruitment and exocytosis are affected in the majority of risk loci. PMID- 19808893 TI - ATP-sensitive K+ channel mediates the zinc switch-off signal for glucagon response during glucose deprivation. AB - OBJECTIVE: The intraislet insulin hypothesis proposes that glucagon secretion during hypoglycemia is triggered by a decrease in intraislet insulin secretion. A more recent hypothesis based on in vivo data from hypoglycemic rats is that it is the decrease in zinc cosecreted with insulin from beta-cells, rather than the decrease in insulin itself, that signals glucagon secretion from alpha-cells during hypoglycemia. These studies were designed to determine whether closure of the alpha-cell ATP-sensitive K(+) channel (K(ATP) channel) is the mechanism through which the zinc switch-off signal triggers glucagon secretion during glucose deprivation. RESEARCH DESIGN AND METHODS: All studies were performed using perifused isolated islets. RESULTS: In control experiments, the expected glucagon response to an endogenous insulin switch-off signal during glucose deprivation was observed in wild-type mouse islets. In experiments with streptozotocin-treated wild-type islets, a glucagon response to an exogenous zinc switch-off signal was observed during glucose deprivation. However, this glucagon response to the zinc switch-off signal during glucose deprivation was not seen in the presence of nifedipine, diazoxide, or tolbutamide or if K(ATP) channel knockout mouse islets were used. All islets had intact glucagon responses to epinephrine. CONCLUSIONS: These data demonstrate that closure of K(ATP) channels and consequent opening of calcium channels is the mechanism through which the zinc switch-off signal triggers glucagon secretion during glucose deprivation. PMID- 19808894 TI - Tpl2 kinase is upregulated in adipose tissue in obesity and may mediate interleukin-1beta and tumor necrosis factor-{alpha} effects on extracellular signal-regulated kinase activation and lipolysis. AB - OBJECTIVE: Activation of extracellular signal-regulated kinase-(ERK)-1/2 by cytokines in adipocytes is involved in the alterations of adipose tissue functions participating in insulin resistance. This study aims at identifying proteins regulating ERK1/2 activity, specifically in response to inflammatory cytokines, to provide new insights into mechanisms leading to abnormal adipose tissue function. RESEARCH DESIGN AND METHODS: Kinase activities were inhibited with pharmacological inhibitors or siRNA. Lipolysis was monitored through glycerol production. Gene expression in adipocytes and adipose tissue of obese mice and subjects was measured by real-time PCR. RESULTS: IkappaB kinase-(IKK) beta inhibition prevented mitogen-activated protein (MAP) kinase kinase (MEK)/ERK1/2 activation in response to interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha but not insulin in 3T3-L1 and human adipocytes, suggesting that IKKbeta regulated a MAP kinase kinase kinase (MAP3K) involved in ERK1/2 activation induced by inflammatory cytokines. We show that the MAP3K8 called Tpl2 was expressed in adipocytes and that IL-1beta and TNF-alpha activated Tpl2 and regulated its expression through an IKKbeta pathway. Pharmacological inhibition or silencing of Tpl2 prevented MEK/ERK1/2 activation by these cytokines but not by insulin, demonstrating its involvement in ERK1/2 activation specifically in response to inflammatory stimuli. Importantly, Tpl2 was implicated in cytokine induced lipolysis and in insulin receptor substrate-1 serine phosphorylation. Tpl2 mRNA expression was upregulated in adipose tissue of obese mice and patients and correlated with TNF-alpha expression. CONCLUSIONS: Tpl2 is selectively involved in inflammatory cytokine-induced ERK1/2 activation in adipocytes and is implicated in their deleterious effects on adipocyte functions. The deregulated expression of Tpl2 in adipose tissue suggests that Tpl2 may be a new actor in adipose tissue dysfunction in obesity. PMID- 19808895 TI - Attenuated purinergic receptor function in patients with type 2 diabetes. AB - OBJECTIVE: Extracellular nucleotides and nucleosides are involved in regulation of skeletal muscle blood flow. Diabetes induces cardiovascular dysregulation, but the extent to which the vasodilatatory capacity of nucleotides and nucleosides is affected in type 2 diabetes is unknown. The present study investigated 1) the vasodilatatory effect of ATP, uridine-triphosphate (UTP), and adenosine (ADO) and 2) the expression and distribution of P2Y(2) and P2X(1) receptors in skeletal muscles of diabetic subjects. RESEARCH DESIGN AND METHODS: In 10 diabetic patients and 10 age-matched control subjects, leg blood flow (LBF) was measured during intrafemoral artery infusion of ATP, UTP, and ADO, eliciting a blood flow equal to knee-extensor exercise at 12 W (approximately 2.6 l/min). RESULTS: The vasodilatatory effect of the purinergic system was 50% lower in the diabetic group as exemplified by an LBF increase of 274 +/- 37 vs. 143 +/- 26 ml/micromol ATP x kg, 494 +/- 80 vs. 234 +/- 39 ml/micromol UTP x kg, and 14.9 +/- 2.7 vs. 7.5 +/- 0.6 ml/micromol ADO x kg in control and diabetic subjects, respectively, thus making the vasodilator potency as follows: UTP control subjects (100) > ATP control subjects (55) > UTP diabetic subjects (47) > ATP diabetic subjects (29) > ADO control subjects (3) > ADO diabetic subjects (1.5). The distribution and mRNA expression of receptors were similar in the two groups. CONCLUSIONS: The vasodilatatory effect of the purinergic system is severely reduced in type 2 diabetic patients. The potency of nucleotides varies with the following rank order: UTP > ATP > ADO. This is not due to alterations in receptor distribution and mRNA expression, but may be due to differences in receptor sensitivity. PMID- 19808896 TI - Grp78 heterozygosity promotes adaptive unfolded protein response and attenuates diet-induced obesity and insulin resistance. AB - OBJECTIVE: To investigate the role of the endoplasmic reticulum (ER) chaperone glucose-regulated protein (GRP) 78/BiP in the pathogenesis of obesity, insulin resistance, and type 2 diabetes. RESEARCH DESIGN AND METHODS: Male Grp78(+/-) mice and their wild-type littermates were subjected to a high-fat diet (HFD) regimen. Pathogenesis of obesity and type 2 diabetes was examined by multiple approaches of metabolic phenotyping. Tissue-specific insulin sensitivity was analyzed by hyperinsulinemic-euglycemic clamps. Molecular mechanism was explored via immunoblotting and tissue culture manipulation. RESULTS: Grp78 heterozygosity increases energy expenditure and attenuates HFD-induced obesity. Grp78(+/-) mice are resistant to diet-induced hyperinsulinemia, liver steatosis, white adipose tissue (WAT) inflammation, and hyperglycemia. Hyperinsulinemic-euglycemic clamp studies revealed that Grp78 heterozygosity improves glucose metabolism independent of adiposity and following an HFD increases insulin sensitivity predominantly in WAT. As mechanistic explanations, Grp78 heterozygosity in WAT under HFD stress promotes adaptive unfolded protein response (UPR), attenuates translational block, and upregulates ER degradation-enhancing alpha-mannosidase like protein (EDEM) and ER chaperones, thus improving ER quality control and folding capacity. Further, overexpression of the active form of ATF6 induces protective UPR and improves insulin signaling upon ER stress. CONCLUSIONS: HFD induced obesity and type 2 diabetes are improved in Grp78(+/-) mice. Adaptive UPR in WAT could contribute to this improvement, linking ER homeostasis to energy balance and glucose metabolism. PMID- 19808897 TI - Adaptive cerebral neovascularization in a model of type 2 diabetes: relevance to focal cerebral ischemia. AB - OBJECTIVE: The effect of diabetes on neovascularization varies between different organ systems. While excessive angiogenesis complicates diabetic retinopathy, impaired neovascularization contributes to coronary and peripheral complications of diabetes. However, how diabetes influences cerebral neovascularization is not clear. Our aim was to determine diabetes-mediated changes in the cerebrovasculature and its impact on the short-term outcome of cerebral ischemia. RESEARCH DESIGN AND METHODS: Angiogenesis (capillary density) and arteriogenesis (number of collaterals and intratree anostomoses) were determined as indexes of neovascularization in the brain of control and type 2 diabetic Goto-Kakizaki (GK) rats. The infarct volume, edema, hemorrhagic transformation, and short-term neurological outcome were assessed after permanent middle-cerebral artery occlusion (MCAO). RESULTS: The number of collaterals between middle and anterior cerebral arteries, the anastomoses within middle-cerebral artery trees, the vessel density, and the level of brain-derived neurotrophic factor were increased in diabetes. Cerebrovascular permeability, matrix metalloproteinase (MMP)-9 protein level, and total MMP activity were augmented while occludin was decreased in isolated cerebrovessels of the GK group. Following permanent MCAO, infarct size was smaller, edema was greater, and there was no macroscopic hemorrhagic transformation in GK rats. CONCLUSIONS: The augmented neovascularization in the GK model includes both angiogenesis and arteriogenesis. While adaptive arteriogenesis of the pial vessels and angiogenesis at the capillary level may contribute to smaller infarction, changes in the tight junction proteins may lead to the greater edema following cerebral ischemia in diabetes. PMID- 19808898 TI - The Pit-1/Pou1f1 transcription factor regulates and correlates with prolactin expression in human breast cell lines and tumors. AB - The transcription factor Pit-1/Pou1f1 regulates GH and prolactin (PRL) secretion in the pituitary gland. Pit-1 expression and GH regulation by Pit-1 have also been demonstrated in mammary gland. However, no data are available on the role of Pit-1 on breast PRL. To evaluate this role, several human breast cancer cell lines were transfected with either the Pit-1 expression vector or a Pit-1 small interference RNA construct, followed by PRL mRNA and protein evaluation. In addition, transient transfection of MCF-7 cells by a reporter construct containing the proximal PRL promoter, and ChIP assays were performed. Our data indicate that Pit-1 regulates mammary PRL at transcriptional level by binding to the proximal PRL promoter. We also found that Pit-1 raises cyclin D1 expression before increasing PRL levels, suggesting a PRL-independent effect of Pit-1 on cell proliferation. By using immunohistochemistry, we found a significant correlation between Pit-1 and PRL expression in 94 human breast invasive ductal carcinomas. Considering the possible role of PRL in breast cancer disorders, the function of Pit-1 in breast should be the focus of further research. PMID- 19808900 TI - Treatment with GH in adults with GH deficiency. Foreword. PMID- 19808899 TI - Expression of hypoxia-inducible factor 1 alpha in thyroid carcinomas. AB - Hypoxia-inducible factor 1 alpha (HIF-1 alpha) is upregulated by hypoxia and oncogenic signalling in many solid tumours. Its regulation and function in thyroid carcinomas are unknown. We evaluated the regulation of HIF-1 alpha and target gene expression in primary thyroid carcinomas and thyroid carcinoma cell lines (BcPAP, WRO, FTC-133 and 8505c). HIF-1 alpha was not detectable in normal tissue but was expressed in thyroid carcinomas. Dedifferentiated anaplastic tumours (ATCs) exhibited high levels of nuclear HIF-1 alpha staining. The HIF-1 target glucose transporter 1 was expressed to a similar level in all tumour types, whereas carbonic anhydrase-9 was significantly elevated in ATCs. In vitro studies revealed a functionally active HIF-1 alpha pathway in thyroid cells with transcriptional activation observed after graded hypoxia (1% O(2), anoxia) or treatment with a hypoxia mimetic cobalt chloride. High basal and hypoxia-induced expression of HIF-1 alpha in FTC-133 cells that harbour a phosphatase and tensin homologue (PTEN) mutation was reduced by introduction of wild-type PTEN. Similarly, pharmacological inhibition of the phosphoinositide 3-kinase (PI3K) pathway using LY294002 inhibited HIF-1 alpha and HIF-1 alpha targets in all cell lines, including those with B-RAF mutations (BcPAP and 8505c). In contrast, the effects of inhibition of the RAF/MEK/extracellular signal-regulated kinase pathway were restricted by environmental condition and B-RAF mutation status. HIF 1 is functionally expressed in thyroid carcinomas and is regulated not only by hypoxia but also via growth factor signalling pathways and, in particular, the PI3K pathway. Given the strong association of HIF-1 alpha with an aggressive disease phenotype and therapeutic resistance, this pathway may be an attractive target for improved therapy in thyroid carcinomas. PMID- 19808901 TI - The PPAR-gamma Pro12Ala polymorphism associates with weight gain during GH treatment in short children born small for gestational age. AB - CONTEXT: Short children born small for gestational age (SGA) have a lean phenotype with lower insulin sensitivity and higher blood pressure. GH treatment results in weight gain, and a decrease in blood pressure and insulin sensitivity. However, not all children respond in the same way. The Pro12Ala polymorphism of the peroxisome proliferator-activated receptor (PPAR-gamma) gene is inversely associated with body mass index (BMI), changes in BMI and the risk to develop type 2 diabetes mellitus. OBJECTIVE: To analyze the contribution of the PPAR gamma Pro12Ala polymorphism to GH induced changes in determinants of metabolic and cardiovascular disease in short SGA children. METHODS: PPAR-gamma was genotyped in 238 Caucasian short SGA children (mean age 7.5 years). Height, weight, blood pressure, and serum lipids were measured before start and during 4 years of GH treatment. In addition, glucose homeostasis by homeostasis model assessment insulin resistance ratio (HOMA-IR) (n=148) and by frequently sampled i.v. glucose tolerance test (n=51), and body composition by dual energy X-ray absorptiometry (n=79) were measured. RESULTS: At baseline, the Ala12 allele was not associated with any determinant of metabolic and cardiovascular disease. After 4 years of GH treatment, the increase in weight for height SDS and BMI SDS was significantly greater in carriers of an Ala12 allele than in noncarriers. The change in all other parameters was not associated with Pro12Ala genotype. CONCLUSION: The Ala12 variant of the PPAR-gamma gene is associated with higher weight gain during GH treatment but not with changes in determinants of metabolic and cardiovascular diseases in Caucasian subjects born SGA. PMID- 19808902 TI - Do we need still more trials on T4 and T3 combination therapy in hypothyroidism? AB - Approximately 10% of hypothyroid patients are dissatisfied with the outcome of levothyroxine replacement. It is unlikely that slight over- or under-treatment with thyroxine (T(4)) explains remaining complaints. Meta-analysis of randomized clinical trials shows no advantage of T(4)/tri-iodothyronine (T(3)) combination therapy over T(4) monotherapy. However, each of these trials can be criticized, and none is perfect: most of them failed to mimic the physiological ratio of serum free T(4) (FT(4)) to free T(3) (FT(3)) concentrations. Development of a sustained-release T(3) preparation given as a single nighttime dose (together with levothyroxine once daily) might maintain physiological serum FT(4)-FT(3) ratio's throughout 24 h. Genetic polymorphisms in deiodinase 2 and thyroid hormone transporters have been associated with well-being, fatigue, depression, and greater improvement on combination therapy. Future trials should target carriers of these polymorphisms to see whether they do better on T(4)/T(3) combination therapy than on T(4) monotherapy. PMID- 19808903 TI - Notch signal activates hypoxia pathway through HES1-dependent SRC/signal transducers and activators of transcription 3 pathway. AB - We report a Notch signal-induced pathway that leads to transcriptional activation of HIF1-alpha gene. HeLa/rtTAA/TRE-N1-IC cell line capable of doxycycline-induced expression of human Notch1-IC was established. The induction of Notch signaling activates HIF1-alpha and its target gene expression in HeLa/rtTAA/TRE-N1-IC cells. Notch signaling enhanced signal transducers and activators of transcription 3 (STAT3) phosphorylation required for HIF1-alpha expression. SRC kinase was found to be responsible for the enhanced STAT3 phosphorylation in response to Notch signaling. Activation of SRC/STAT3 pathway by Notch signaling was dependent on the expression of Notch effector HES1 transcription factor. The induction of HES1 enhanced STAT3 phosphorylation at Tyr 705 as well as SRC phosphorylation at Tyr 416 in inducible HeLa/rtTAA/TRE-HES1 cells, which express HES1 in response to doxycycline treatment. However, the treatment of Trichostatin A that interferes with HES1 transcriptional regulation did not affect STAT3 phosphorylation, and the expression of dominant negative HES1 failed to interfere with HES1-dependent SRC/STAT3 pathway. These observations have led us to the conclusion that HES1-dependent activation of SRC/STAT3 pathway is independent of HES1 transcription regulation. This study first reports HES1-dependent SRC/STAT3 pathway that provides a functional link between Notch signaling and hypoxia pathway. PMID- 19808904 TI - The role of MET activation in determining the sensitivity to epidermal growth factor receptor tyrosine kinase inhibitors. AB - The development of resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKI) seems almost inevitable, even in patients with lung cancer that initially respond well to EGFR-TKIs. MET amplification was recently found to be a mechanism of escape from the anticancer effect of EGFR inhibitors. In the present study, we investigated the means whereby MET affects sensitivity to EGFR TKIs in PC-9 cells. Gefitinib- or erlotinib-resistant sublines were established by exposing the parental PC-9 cell line to chronic, repeated treatments with these drugs. These resistant sublines showed more than 100-fold more resistance to gefitinib and erlotinib and acquired cross-resistance to other EGFR-TKIs. The T790M EGFR mutation was found by pyrosequencing, and this seemed to be the cause of drug resistance. Resistant cells also showed MET activation, although gene amplification was not detected. Furthermore, the induction of MET activity was not found to be associated with sensitivity to EGFR-TKIs. Interestingly, increased passage number without exposure to gefitinib or erlotinib caused MET activation, but this did not affect sensitivity to EGFR-TKIs. In addition, hepatocyte growth factor was found to block the ability of EGFR-TKIs to inhibit MET activation. However, sustained MET activation by hepatocyte growth factor did not modulate the cellular effects of gefitinib or erlotinib. Rather, activated MET enhanced migration and invasion abilities. Summarizing, MET activation may be acquired during cancer cell proliferation and enhances migratory and invasive abilities without affecting cellular sensitivity to EGFR-TKIs. Accordingly, the present study suggests that MET activation caused by factors other than MET gene amplification is not a suitable surrogate marker of resistance to EGFR-TKIs. PMID- 19808905 TI - Integrin (alpha6beta4) signals through Src to increase expression of S100A4, a metastasis-promoting factor: implications for cancer cell invasion. AB - Integrin alpha6beta4 is linked to cancer cell motility and invasion in aggressive and metastatic cancer cells. In this study, we showed that expression of the beta4 integrin in MDA-MB-435 cancer cells (MDA-MB-435/beta4) leads to a dramatic increase in expression of a metastasis-promoting factor, S100A4, as determined by affymetrix gene chip microarray, quantitative real-time PCR, and Western blot analysis. Alternatively, knocking down beta4 integrin expression in MDA-MB-231 breast carcinoma cells by shRNA reduced the level of S100A4 expression. The mechanism by which alpha6beta4 enhances S100A4 expression involves Src, Akt, and NFAT. We have further shown that Y1494, a tyrosine residue of the ITIM motif in the cytoplasmic domain of the beta4 integrin subunit, is essential for alpha6beta4-dependent S100A4 expression. Reduction of S100A4 expression by shRNA blocked migration, invasion, and anchorage-independent growth of MDA-MB 435/beta4, SUM-159, and MDA-MB-231 cells. These studies define a novel mechanism by which integrin alpha6beta4 promotes cancer cell motility and invasion, and provides insight into how S100A4 expression is regulated in cancer cells. PMID- 19808906 TI - Promyelocytic leukemia nuclear bodies link the DNA damage repair pathway with hepatitis B virus replication: implications for hepatitis B virus exacerbation during chemotherapy and radiotherapy. AB - The mechanism responsible for hepatitis B virus (HBV) exacerbation during chemotherapy and radiotherapy remains unknown. We investigated whether the activation of DNA repair pathways influences HBV replication. The upregulation of the promyelocytic leukemia (PML) protein and its associated PML nuclear body (PML NB) by chemotherapy and irradiation-induced DNA repair signaling correlated with the upregulation of HBV pregenomic transcription, HBV-core expression, and HBV DNA replication. The HBV-core protein and HBV DNA localized to PML-NBs, where they associated with PML and histone deacetylase 1 (HDAC1). Chemotherapy and radiotherapy affected the interactions between PML, HBV-core, and HDAC1. The enhanced protein-protein interaction between PML and HBV-core inhibited PML mediated apoptosis and decreased PML-associated HDAC activity. The reversal of HDAC-mediated repression on the HBV covalently closed circular DNA basal core promoter resulted in the amplification of HBV-core and pregenomic expression. These results suggest that PML in PML-NBs links the DNA damage response with HBV replication and may cooperate with HBV-core and HDAC1 on the HBV covalently closed circular DNA basal core promoter to form a positive feedback loop for HBV exacerbation during chemotherapy and radiotherapy. PMID- 19808907 TI - Activation of sodium-glucose cotransporter 1 ameliorates hyperglycemia by mediating incretin secretion in mice. AB - Glucose ingestion stimulates the secretion of the incretin hormones, glucose dependent insulinotropic peptide (GIP) and glucagon-like peptide-1 (GLP-1). Despite the critical role of incretins in glucose homeostasis, the mechanism of glucose-induced incretin secretion has not been established. We investigated the underlying mechanism of glucose-induced incretin secretion in vivo in mice. Injection of glucose at 1 g/kg in the upper intestine significantly increased plasma GIP and GLP-1 levels, whereas injection of glucose in the colon did not increase GIP or GLP-1 levels. This finding indicates that the glucose sensor for glucose-induced incretin secretion is in the upper intestine. Coadministration of a sodium-glucose cotransporter-1 (SGLT1) inhibitor, phloridzin, with glucose in the upper intestine blocked glucose absorption and glucose-induced incretin secretion. alpha-methyl-d-glucopyranoside (MDG), an SGLT1 substrate that is a nonmetabolizable sugar, significantly increased plasma GIP and GLP-1 levels, whereas phloridzin blocked these increases, indicating that concomitant transport of sodium ions and glucose (substrate) via SGLT1 itself triggers incretin secretion without the need for subsequent glucose metabolism. Interestingly, oral administration of MDG significantly increased plasma GIP, GLP-1, and insulin levels and reduced blood glucose levels during an intraperitoneal glucose tolerance test. Furthermore, chronic MDG treatment in drinking water (3%) for 13 days reduced blood glucose levels after a 2-h fast and in an oral glucose tolerance test in diabetic db/db mice. Our findings indicate that SGLT1 serves as the intestinal glucose sensor for glucose-induced incretin secretion and that a noncalorigenic SGLT1 substrate ameliorates hyperglycemia by stimulating incretin secretion. PMID- 19808908 TI - Quantification of pancreatic islet distribution in situ in mice. AB - Tracing changes of specific cell populations in health and disease is an important goal of biomedical research. Precisely monitoring pancreatic beta-cell proliferation and islet growth is a challenging area of research. We have developed a method to capture the distribution of beta-cells in the intact pancreas of transgenic mice with fluorescence-tagged beta-cells with a macro written for ImageJ (rsb.info.nih.gov/ij/). Total beta-cell area and islet number and size distribution are quantified with reference to specific parameters and location for each islet and for small clusters of beta-cells. The entire distribution of islets can now be plotted in three dimensions, and the information from the distribution on the size and shape of each islet allows a quantitative and a qualitative comparison of changes in overall beta-cell area at a glance. PMID- 19808909 TI - Pigment epithelium-derived factor suppresses adipogenesis via inhibition of the MAPK/ERK pathway in 3T3-L1 preadipocytes. AB - We previously reported that circulating levels of pigment epithelium-derived factor (PEDF), a newly identified adipokine, are increased in patients with type 2 diabetes, correlating with body mass index. However, the role of PEDF in adipogenesis remains elusive. In the present study, we have investigated the effects and mechanisms of PEDF on adipocyte differentiation in 3T3-L1 preadipocytes. Differentiation of 3T3-L1 preadipocytes was induced in the presence or absence of human recombinant PEDF protein. The effects of PEDF on adipogenic gene expression, mitotic clonal expansion (MCE), and MAPK activation were investigated. Physiological concentrations of human PEDF protein inhibited adipocyte differentiation, evidenced by decreased lipid accumulation, downregulation of adipocyte markers, and inhibition of master adipogenic transcription factors such as C/EBP-alpha and PPARgamma. The antiadipogenic effects of PEDF were observed only when PEDF was added to the cells on day 0, but not on day 3 during differentiation, suggesting that PEDF targets some early adipogenic events. Similarly, overexpression of PEDF by adenovirus attenuated adipocyte differentiation. Further studies revealed that PEDF, or U-0126, a specific MAPK/ERK inhibitor, sequentially inhibited the early activation of ERK and MCE. Moreover, PEDF attenuated expression and the phosphorylation of C/EBP beta at Thr(188), an essential step for transcriptional activation of C/EBP-beta. In addition, PEDF expression was decreased significantly in the first 24 h during adipocyte differentiation, suggesting that downregulation of PEDF may be essential for the initiation of MCE and adipogenesis. We conclude that PEDF inhibits adipogenesis in 3T3-L1 preadipocytes partially because of inhibition of the MAPK/ERK signaling pathway and MCE. PMID- 19808910 TI - Chronically elevated plasma C-type natriuretic peptide level stimulates skeletal growth in transgenic mice. AB - C-type natriuretic peptide (CNP) plays a critical role in endochondral ossification through guanylyl cyclase-B (GC-B), a natriuretic peptide receptor subtype. Cartilage-specific overexpression of CNP enhances skeletal growth and rescues the dwarfism in a transgenic achondroplasia model with constitutive active mutation of fibroblast growth factor receptor-3. For future clinical application, the efficacy of CNP administration on skeletal growth must be evaluated. Due to the high clearance of CNP, maintaining a high concentration is technically difficult. However, to model high blood CNP concentration, we established a liver-targeted CNP-overexpressing transgenic mouse (SAP-CNP tgm). SAP-CNP tgm exhibited skeletal overgrowth in proportion to the blood CNP concentration and revealed phenotypes of systemic stimulation of cartilage bones, including limbs, paws, costal bones, spine, and skull. Furthermore, in SAP-CNP tgm, the size of the foramen magnum, the insufficient formation of which results in cervico-medullary compression in achondroplasia, also showed significant increase. CNP primarily activates GC-B, but under high concentrations it cross reacts with guanylyl cyclase-A (GC-A), a natriuretic peptide receptor subtype of atrial natriuretic peptides (ANP) and brain natriuretic peptides (BNP). Although activation of GC-A could alter cardiovascular homeostasis, leading to hypotension and heart weight reduction, the skeletal overgrowth phenotype in the line of SAP CNP tgm with mild overexpression of CNP did not accompany decrease of systolic blood pressure or heart weight. These results suggest that CNP administration stimulates skeletal growth without adverse cardiovascular effect, and thus CNP could be a promising remedy targeting achondroplasia. PMID- 19808911 TI - Rosiglitazone decreases C-reactive protein to a greater extent relative to glyburide and metformin over 4 years despite greater weight gain: observations from a Diabetes Outcome Progression Trial (ADOPT). AB - OBJECTIVE: C-reactive protein (CRP) is closely associated with obesity and cardiovascular disease in both diabetic and nondiabetic populations. In the short term, commonly prescribed antidiabetic agents have different effects on CRP; however, the long-term effects of those agents are unknown. RESEARCH DESIGN AND METHODS: In A Diabetes Outcome Progression Trial (ADOPT), we examined the long term effects of rosiglitazone, glyburide, and metformin on CRP and the relationship among CRP, weight, and glycemic variables in 904 subjects over 4 years. RESULTS: Baseline CRP was significantly correlated with homeostasis model assessment of insulin resistance (HOMA-IR), A1C, BMI, waist circumference, and waist-to-hip ratio. CRP reduction was greater in the rosiglitazone group by 47.6% relative to glyburide and by -30.5% relative to metformin at 48 months. Mean weight gain from baseline (at 48 months) was 5.6 kg with rosiglitazone, 1.8 kg with glyburide, and -2.8 kg with metformin. The change in CRP from baseline to 12 months was correlated positively with change in BMI in glyburide (r = 0.18) and metformin (r = 0.20) groups but not in the rosiglitazone (r = -0.05, NS) group. However, there was no longer a significant correlation between change in CRP and change in HOMA-IR, A1C, or waist-to-hip ratio in any of the three treatment groups. CONCLUSIONS: Rosiglitazone treatment was associated with durable reductions in CRP independent of changes in insulin sensitivity, A1C, and weight gain. CRP in the glyburide and metformin groups was positively associated with changes in weight, but this was not the case with rosiglitazone. PMID- 19808912 TI - Randomized placebo-controlled double-blind clinical trial of cannabis-based medicinal product (Sativex) in painful diabetic neuropathy: depression is a major confounding factor. AB - OBJECTIVE: To assess the efficacy of Sativex, a cannabis-based medicinal extract, as adjuvant treatment in painful diabetic peripheral neuropathy (DPN). RESEARCH DESIGN AND METHODS: In this randomized controlled trial, 30 subjects with painful DPN received daily Sativex or placebo. The primary outcome measure was change in mean daily pain scores, and secondary outcome measures included quality-of-life assessments. RESULTS: There was significant improvement in pain scores in both groups, but mean change between groups was not significant. There were no significant differences in secondary outcome measures. Patients with depression had significantly greater baseline pain scores that improved regardless of intervention. CONCLUSIONS: This first-ever trial assessing the efficacy of cannabis has shown it to be no more efficacious than placebo in painful DPN. Depression was a major confounder and may have important implications for future trials on painful DPN. PMID- 19808913 TI - Postprandial vascular effects of VIAject compared with insulin lispro and regular human insulin in patients with type 2 diabetes. AB - OBJECTIVE: Recent studies suggested an impact of prandial insulin delivery on postprandial regulation of tissue blood flow. This study compared the effect of VIAject with human regular insulin and insulin lispro on postprandial oxidative stress and endothelial function in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: Fourteen patients (seven men; aged 61.5 +/- 1.8 years; duration of diabetes 6.6 +/- 4.6 years; A1C 7.2 +/- 0.5% [mean +/- SEM]) received a prandial injection of VIAject, human regular insulin, and insulin lispro. At baseline and after a standardized liquid meal test (Ensure Plus), the postprandial increases in asymmetric dimethylarginine (ADMA) and nitrotyrosine levels were investigated. In addition, the postprandial effects on microvascular blood flow, skin oxygenation, and vascular elasticity were measured. RESULTS: Treatment with VIAject resulted in a significant reduction in the peak postprandial generation of ADMA compared with human insulin and insulin lispro (VIAject -27.3 +/- 22.6, human insulin 97.7 +/- 24.4, and insulin lispro 66.9 +/- 33.9 nmol/l; P < 0.05, respectively). The postprandial increases in nitrotyrosine levels were significantly less after VIAject than after human regular insulin (VIAject -0.22 +/- 0.17 vs. human insulin 0.25 +/- 0.15 microg/ml; P < 0.05), whereas nitrotyrosine after insulin lispro was in between (insulin lispro 0.09 +/- 0.07 microg/ml; NS). In parallel, earlier and more pronounced increases in microvascular blood flow and skin oxygenation were obtained after VIAject compared with those after human insulin or insulin lispro (P < 0.05, respectively). All insulin formulations resulted in comparable improvements in central arterial elasticity. CONCLUSIONS; Treatment with VIAject reduced postprandial oxidative stress and improved endothelial function compared with human regular insulin or insulin lispro. PMID- 19808914 TI - Insulin glargine safety in pregnancy: a transplacental transfer study. AB - OBJECTIVE: Insulin glargine (Lantus) is an extended-action insulin analog with greater stability and duration of action than regular human insulin. The long duration of action and decreased incidence of hypoglycemia provide potential advantages for its use in pregnancy. However, the placental pharmacokinetics of insulin glargine have not been studied. Therefore, the objective of this study was to determine whether insulin glargine crosses the human placenta using the human perfused placental lobule technique. RESEARCH DESIGN AND METHODS: Placentae were obtained with informed consent after elective cesarean section delivery of noncomplicated term pregnancies. Insulin glargine, at a therapeutic concentration of 150 pmol/l (20 microU/ml) was added to the maternal circulation. Additional experiments were carried out at insulin glargine concentrations 1,000-fold higher than therapeutic levels (150, 225, and 300 nmol/l). A subsequent perfusion for which the maternal circuit remained open and insulin glargine was continuously infused at 150 pmol/l was completed for further confirmation of findings. The appearance of insulin glargine in the fetal circulation was analyzed by a chemiluminescence immunoassay. RESULTS: Results from perfusions carried out at therapeutic concentrations (150 pmol/l) of insulin glargine showed no detectable insulin glargine in the fetal circuit. After perfusion with very high insulin glargine concentrations of 150, 225, and 300 nmol/l, the rate of transfer remained low at 0.079 +/- 0.01, 0.14, and 0.064 pmol . min(-1) . g tissue(-1), respectively. CONCLUSIONS: Insulin glargine, when used at therapeutic concentrations, is not likely to cross the placenta. PMID- 19808915 TI - Relationship between carotid intima-media thickness and silent cerebral infarction in Japanese subjects with type 2 diabetes. AB - OBJECTIVE: We examined the relationship between intima-media thickness of common carotid artery (CCA-IMT) and silent cerebral infarction (SCI) with the magnetic resonance imaging (MRI) study in Japanese subjects with type 2 diabetes. RESEARCH DESIGN AND METHODS: The brain MRI study and the carotid ultrasonography were performed in a total of 217 consecutive Japanese subjects with type 2 diabetes. Various risk factors for SCI were examined using multiple logistic analyses. RESULTS: The SCI was found in 60.4% of the diabetic subjects. In the diabetic subjects, age, systolic blood pressure (SBP), pulse wave velocity, and CCA-IMT were significantly higher in the subjects with SCI than in those without it. Multiple logistic analyses indicated that age, SBP, and CCA-IMT were significant and independent risk factors of SCI in the diabetic subjects. CONCLUSIONS: CCA IMT, but not pulse wave velocity, was independently associated with SCI in Japanese subjects with type 2 diabetes. PMID- 19808916 TI - Psychometric findings for a Spanish translation of the diabetes self-management profile (DSMP-Parent-Sp). AB - OBJECTIVE: Few validated measures exist to evaluate self-management of diabetes in families with limited English proficiency. The present study evaluated the psychometric properties and the factorial equivalence of a Spanish translation of the parent report version of the Diabetes Self-Management Profile (DSMP-Parent Sp). RESEARCH DESIGN AND METHODS: Hispanic families of youth (mean 13.7 years old) with type 1 diabetes were recruited from three clinics in South Florida and represented a wide range of nationalities and acculturation levels. A total of 127 parents reported on their child's self-management behaviors using either the original DSMP-Parent (59.8%) or the DSMP-Parent-Sp (40.2%). In addition, youth reported their self-management using the original DSMP in English, and physicians rated their perceptions of the youth's self-management. Glycemic control was indexed by A1C in the past 3 months and collected from medical chart review. RESULTS: Item analysis confirmed that the DSMP-Parent-Sp items related to the overall composite score in expected ways, and internal consistency estimates were adequate. Paired correlations demonstrated strong parent-child concordance and a significant relationship with physician perceptions of self-management. Evidence of concurrent and convergent validity, as well as "strict factorial invariance," was demonstrated. CONCLUSIONS: These preliminary findings indicate that the DSMP Parent-Sp is a reliable and valid parent report measure of the diabetes self management behaviors of Hispanic youths. In addition, there is preliminary evidence that the translated measure may be considered equivalent to the original English measure when used to measure self-management in Hispanic youth with diabetes. PMID- 19808917 TI - A1C is associated with intima-media thickness in individuals with normal glucose tolerance. AB - OBJECTIVE: One-hour glucose during an oral glucose tolerance test (OGTT) was recently proposed as a valuable marker to identify individuals with normal glucose tolerance (NGT) and increased intima-media thickness (IMT). However, central markers of glycemic control were not considered. The aim of this study was to identify which marker of glycemic control is most informative with respect to the variation of IMT in individuals with NGT. RESEARCH DESIGN AND METHODS: Cardiovascular risk factors, glucose metabolism (OGTT), and IMT were determined in 1,219 nondiabetic individuals (851 women, 368 men; 558 with NGT). RESULTS: One hour glucose and A1C levels were significantly correlated to carotid IMT in individuals with NGT, whereas fasting and 2-h glucose levels were not informative. Only A1C was associated with IMT independent of other confounders, whereas 1-h glucose was not informative. Comparable results were found in the total cohort, including individuals with IFG and IGT. CONCLUSIONS: A1C was the most informative glycemic marker with respect to IMT in individuals with NGT. PMID- 19808918 TI - Patients diagnosed with diabetes are at increased risk for asthma, chronic obstructive pulmonary disease, pulmonary fibrosis, and pneumonia but not lung cancer. AB - OBJECTIVE: There are limited data on the risk of pulmonary disease in patients with diabetes. The aim of this study was to evaluate and compare the incidence of asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pneumonia, and lung cancer in patients with and without a diagnosis of diabetes. RESEARCH DESIGN AND METHODS: We conducted a retrospective, longitudinal cohort study using the electronic records of a large health plan in northern California. Age and sex data were available for all cohort members (n = 1,811,228). Data on confounders were available for a subcohort that responded to surveys (n = 121,886), among whom Cox proportional hazards regression models were fit. RESULTS: Age- and sex-adjusted incidence rates and 95% CIs were calculated for members with and without diabetes in the full cohort and the subcohort. No difference was observed for lung cancer, but the incidence of asthma, COPD, fibrosis, and pneumonia was significantly higher in those members with a diagnosis of diabetes. These differences remained significant in regression models adjusted for age, sex, race/ethnicity, smoking, BMI, education, alcohol consumption, and outpatient visits (asthma hazard ratio [HR] 1.08 [95% CI 1.03 1.12], COPD HR 1.22 [1.15-1.28], pulmonary fibrosis HR 1.54 [1.31-1.81], and pneumonia HR 1.92 [1.84-1.99]). The risk of pneumonia and COPD increased significantly with increasing A1C. CONCLUSIONS: Individuals with diabetes are at increased risk of several pulmonary conditions (asthma, COPD, fibrosis, and pneumonia) but not lung cancer. This increased risk may be a consequence of declining lung function in patients with diabetes. PMID- 19808919 TI - Insulin resistance, beta-cell dysfunction, and conversion to type 2 diabetes in a multiethnic population: the Insulin Resistance Atherosclerosis Study. AB - OBJECTIVE: Insulin resistance and beta-cell function are major predictors of type 2 diabetes, but studies using direct methods of insulin resistance and secretion are few and relatively small. Furthermore, the strength of these associations has not been tested in different ethnic groups and various states of glucose tolerance, family history of diabetes, and obesity. RESEARCH DESIGN AND METHODS: Predictors of incident diabetes were evaluated in Hispanic, non-Hispanic white, and African American participants in the Insulin Resistance Atherosclerosis Study (aged 40-69 years). In 557 participants with normal glucose tolerance and 269 with impaired glucose tolerance (IGT), insulin sensitivity (insulin sensitivity index [S(I)]) and first-phase insulin secretion (acute insulin response [AIR]) were directly measured using the frequently sampled intravenous glucose tolerance test. RESULTS: At the 5-year follow-up examination, 128 (15.5%) individuals had developed diabetes. Both S(I) (odds ratio x 1 SD 0.50 [95% CI 0.37-0.68]) and AIR (0.51 [0.40-0.65]) were independent predictors of incident diabetes even after adjustment for age, sex, ethnicity, center, IGT, family history of diabetes, and BMI. The strength of the relation of S(I) and AIR to incident diabetes was not significantly affected by potential interactions of age, sex, ethnicity, glucose tolerance, BMI, or family history of diabetes (P > or = 0.15). CONCLUSIONS: Both insulin sensitivity and beta-cell function predict conversion to diabetes in different ethnic groups and various states of glucose tolerance, family history of diabetes, and obesity. The prevention of type 2 diabetes should focus on interventions that improve both insulin resistance and insulin secretion. PMID- 19808920 TI - Comparison of A1C and fasting glucose criteria to diagnose diabetes among U.S. adults. AB - OBJECTIVE: To compare A1C and fasting glucose for the diagnosis of diabetes among U.S. adults. RESEARCH DESIGN AND METHODS: This study included 6,890 adults (> or =20 years of age) from the 1999-2006 National Health and Nutrition Examination Survey without a self-reported history of diabetes who had fasted > or =9 h. A1C > or =6.5% and fasting glucose > or =126 mg/dl were used, separately, to define diabetes. RESULTS: Overall, 1.8% of U.S. adults had A1C > or =6.5% and fasting glucose > or =126 mg/dl, 0.5% had A1C > or =6.5% and fasting glucose <126 mg/dl, and 1.8% had A1C <6.5% and fasting glucose > or =126 mg/dl. Compared with individuals with A1C <6.5% and fasting glucose > or =126 mg/dl, individuals with A1C > or =6.5% and fasting glucose <126 mg/dl were younger, more likely to be non Hispanic black, had lower Hb levels, and had higher C-reactive protein. CONCLUSIONS: A1C > or =6.5% demonstrates reasonable agreement with fasting glucose for diagnosing diabetes among U.S. adults. PMID- 19808921 TI - Ingestion of diet soda before a glucose load augments glucagon-like peptide-1 secretion. AB - OBJECTIVE: The goal of this study was to determine the effect of artificial sweeteners on glucose, insulin, and glucagon-like peptide (GLP)-1 in humans. RESEARCH DESIGN AND METHODS: For this study, 22 healthy volunteers (mean age 18.5 +/- 4.2 years) underwent two 75-g oral glucose tolerance tests with frequent measurements of glucose, insulin, and GLP-1 for 180 min. Subjects drank 240 ml of diet soda or carbonated water, in randomized order, 10 min prior to the glucose load. RESULTS: Glucose excursions were similar after ingestion of carbonated water and diet soda. Serum insulin levels tended to be higher after diet soda, without statistical significance. GLP-1 peak and area under the curve (AUC) were significantly higher with diet soda (AUC 24.0 +/- 15.2 pmol/l per 180 min) versus carbonated water (AUC 16.2 +/- 9.0 pmol/l per 180 min; P = 0.003). CONCLUSIONS: Artificial sweeteners synergize with glucose to enhance GLP-1 release in humans. This increase in GLP-1 secretion may be mediated via stimulation of sweet-taste receptors on L-cells by artificial sweetener. PMID- 19808922 TI - Validation of the Chinese version of the Problem Areas in Diabetes (PAID-C) scale. AB - OBJECTIVE: To examine the psychometric properties of a Chinese version of the Problem Areas In Diabetes (PAID-C) scale. RESEARCH DESIGN AND METHODS: The reliability and validity of the PAID-C were evaluated in a convenience sample of 205 outpatients with type 2 diabetes. Confirmatory factor analysis, Bland-Altman analysis, and Spearman's correlations facilitated the psychometric evaluation. RESULTS: Confirmatory factor analysis confirmed a one-factor structure of the PAID-C (chi(2)/df ratio = 1.894, goodness-of-fit index = 0.901, comparative fit index = 0.905, root mean square error of approximation = 0.066). The PAID-C was associated with A1C (r(s) = 0.15; P < 0.05) and diabetes self-care behaviors in general diet (r(s) = -0.17; P < 0.05) and exercise (r(s) = -0.17; P < 0.05). The 4-week test-retest reliability demonstrated satisfactory stability (r(s) = 0.83; P < 0.01). CONCLUSIONS: The PAID-C is a reliable and valid measure to determine diabetes-related emotional distress in Chinese people with type 2 diabetes. PMID- 19808923 TI - Zinc transporter 8 antibodies complement GAD and IA-2 antibodies in the identification and characterization of adult-onset autoimmune diabetes: Non Insulin Requiring Autoimmune Diabetes (NIRAD) 4. AB - OBJECTIVE: Zinc transporter 8 (ZnT8) is an islet beta-cell secretory granule membrane protein recently identified as an autoantibody antigen in type 1 diabetes. The aim of this study was to determine the prevalence and role of antibodies to ZnT8 (ZnT8As) in adult-onset diabetes. RESEARCH DESIGN AND METHODS: ZnT8As were measured by a radioimmunoprecipitation assay using recombinant ZnT8 COOH-terminal or NH(2)-terminal proteins in 193 patients with adult-onset autoimmune diabetes having antibodies to either GAD (GADAs) or IA-2 (IA-2As) and in 1,056 antibody-negative patients with type 2 diabetes from the Non Insulin Requiring Autoimmune Diabetes (NIRAD) study. RESULTS: ZnT8As-COOH were detected in 18.6% patients with autoimmune diabetes and 1.4% with type 2 diabetes. ZnT8As NH(2) were rare. ZnT8As were associated with younger age and a high GADA titer. The use of GADAs, IA-2As, and ZnT8As in combination allowed a stratification of clinical phenotype, with younger age of onset of diabetes and characteristics of more severe insulin deficiency (higher fasting glucose and A1C, lower BMI, total cholesterol, and triglycerides) in patients with all three markers, with progressive attenuation in patients with two, one, and no antibodies (all P(trend) < 0.001). Autoantibody titers, association with high-risk HLA genotypes, and prevalence of thyroid peroxidase antibodies followed the same trend (all P < 0.001). CONCLUSIONS: ZnT8As are detectable in a proportion of patients with adult onset autoimmune diabetes and seem to be a valuable marker to differentiate clinical phenotypes. PMID- 19808925 TI - Osteocalcin is related to enhanced insulin secretion in gestational diabetes mellitus. AB - OBJECTIVE: There is growing evidence that osteocalcin, an osteoblast-derived protein locally acting on bone formation, can increase insulin secretion as well as insulin sensitivity and thus prevent the development of obesity and diabetes in experimental animals. In humans, osteocalcin has been reported to be decreased in patients with type 2 diabetes. Because gestational diabetes mellitus (GDM) can serve as a model of pre-type 2 diabetes, the aim of this study was to investigate osteocalcin in GDM. RESEARCH DESIGN AND METHODS: Osteocalcin measurement and an oral glucose tolerance test were performed in 78 pregnant women (26 women had GDM and 52 women had normal glucose tolerance [NGT] during pregnancy; women were matched for age and BMI) and in 34 women postpartum. RESULTS: During pregnancy osteocalcin was significantly higher in the women with GDM than in the women with NGT (15.6 +/- 6.4 vs. 12.6 +/- 4.0 ng/ml; P < 0.015), whereas no difference was observed between the two groups at 12 weeks postpartum (36.2 +/- 10.2 vs. 36.2 +/ 13.0 ng/ml), when osteocalcin was found to be increased compared with the level in the pregnant state in all women (+145 +/- 102% in GDM vs. +187 +/- 119% in NGT; P < 0.0001). Moreover, osteocalcin showed a significant correlation with basal and total insulin secretion in the whole study group (R = 0.3, P < 0.01). CONCLUSIONS: In GDM osteocalcin was higher and thus less restrained than in women with NGT during pregnancy and furthermore correlated with insulin secretion parameters. Therefore, it could be hypothesized that osteocalcin can enhance insulin secretion in insulin-resistant states; alternatively an effect of hyperinsulinemia on osteocalcin secretion cannot be excluded. PMID- 19808924 TI - Effects of exenatide alone and in combination with daclizumab on beta-cell function in long-standing type 1 diabetes. AB - OBJECTIVE: In patients with long-standing type 1 diabetes, we investigated whether improved beta-cell function can be achieved by combining intensive insulin therapy with agents that may 1) promote beta-cell growth and/or limit beta-cell apoptosis and 2) weaken the anti-beta-cell autoimmunity. RESEARCH DESIGN AND METHODS: For this study, 20 individuals (mean age 39.5 +/- 11.1 years) with long-standing type 1 diabetes (21.3 +/- 10.7 years) were enrolled in this prospective open-label crossover trial. After achieving optimal blood glucose control, 16 subjects were randomized to exenatide with or without daclizumab. Endogenous insulin production was determined by repeatedly measuring serum C peptide. RESULTS: In 85% of individuals with long-standing type 1 diabetes who were screened for participation in this trial, C-peptide levels >or=0.05 ng/ml (0.02 nmol/l) were found. Residual beta-cells responded to physiological (mixed meal) and pharmacological (arginine) stimuli. During exenatide treatment, patients lost 4.1 +/- 2.9 kg body wt and insulin requirements declined significantly (total daily dose on exenatide 0.48 +/- 0.11 vs. 0.55 +/- 0.13 units x kg(-1) x day(-1) without exenatide; P = 0.0062). No signs of further activation of the underlying autoimmune disease were observed. Exenatide delayed gastric emptying, suppressed endogenous incretin levels, but did not increase C peptide secretion. CONCLUSIONS: In long-standing type 1 diabetes, which remains an active autoimmune disease even decades after its onset, surviving beta-cells secrete insulin in a physiologically regulated manner. However, the combination of intensified insulin therapy, exenatide, and daclizumab did not induce improved function of these remaining beta-cells. PMID- 19808926 TI - Diabetes Interactive Diary: a new telemedicine system enabling flexible diet and insulin therapy while improving quality of life: an open-label, international, multicenter, randomized study. AB - OBJECTIVE: Widespread use of carbohydrate counting is limited by its complex education. In this study we compared a Diabetes Interactive Diary (DID) with standard carbohydrate counting in terms of metabolic and weight control, time required for education, quality of life, and treatment satisfaction. RESEARCH DESIGN AND METHODS: Adults with type 1 diabetes were randomly assigned to DID (group A, n = 67) or standard education (group B, n = 63) and followed for 6 months. A subgroup also completed the SF-36 Health Survey (SF-36) and World Health Organization-Diabetes Treatment Satisfaction Questionnaire (WHO-DTSQ) at each visit. RESULTS: Of 130 patients (aged 35.7 +/- 9.4 years; diabetes duration 16.5 +/- 10.5 years), 11 dropped out. Time for education was 6 h (range 2-15 h) in group A and 12 h (2.5-25 h) in group B (P = 0.07). A1C reduction was similar in both groups (group A from 8.2 +/- 0.8 to 7.8 +/- 0.8% and group B from 8.4 +/- 0.7 to 7.9 +/- 1.1%; P = 0.68). Nonsignificant differences in favor of group A were documented for fasting blood glucose and body weight. No severe hypoglycemic episode occurred. WHO-DTSQ scores increased significantly more in group A (from 26.7 +/- 4.4 to 30.3 +/- 4.5) than in group B (from 27.5 +/- 4.8 to 28.6 +/- 5.1) (P = 0.04). Role Physical, General Health, Vitality, and Role Emotional SF-36 scores improved significantly more in group A than in group B. CONCLUSIONS: DID is at least as effective as traditional carbohydrate counting education, allowing dietary freedom for a larger proportion of type 1 diabetic patients. DID is safe, requires less time for education, and is associated with lower weight gain. DID significantly improved treatment satisfaction and several quality-of-life dimensions. PMID- 19808927 TI - Utility of homeostasis model assessment of beta-cell function in predicting diabetes in 12,924 healthy Koreans. AB - OBJECTIVE: It is unclear how well homeostasis model assessment of beta-cell function (HOMA-beta) predicts diabetes development beyond its components, especially glucose. RESEARCH DESIGN AND METHODS: We identified 12,924 nondiabetic Koreans who had fasting plasma glucose and insulin concentrations measured in 2003 and again in 2008. To minimize the impact of differences in baseline glucose concentration, individuals were divided into three glucose categories: normal fasting glucose (NFG, glucose <5.6 mmol/l), impaired fasting glucose (IFG-100) (5.6-6.0 mmol/l), and IFG-110 (6.1-6.9 mmol/l). RESULTS: Diabetes developed in 29% of individuals in the IFG-110 group, compared with 5% in IFG-100 and 0.3% in NFG groups. Within each glucose category, those who progressed to diabetes had higher baseline glucose concentrations (P < or = 0.04). Baseline HOMA-beta, however, was not lower but higher in individuals who developed diabetes in the NFG group (P = 0.009) and similar in the IFG-100 and IFG-110 groups. CONCLUSIONS: These data question the utility of using HOMA-beta to predict the development of diabetes. PMID- 19808928 TI - Relationship between A1C and glucose levels in the general Dutch population: the new Hoorn study. AB - OBJECTIVE: To investigate the relationship among A1C, fasting plasma glucose (FPG), and 2-h postload plasma glucose in the Dutch general population and to evaluate the results of using A1C for screening and diagnosis of diabetes. RESEARCH DESIGN AND METHODS: In 2006-2007, 2,753 participants of the New Hoorn Study, aged 40-65 years, who were randomly selected from the population of Hoorn, the Netherlands, underwent an oral glucose tolerance test (OGTT). Glucose status (normal glucose metabolism [NGM], intermediate hyperglycemia, newly diagnosed diabetes, and known diabetes) was defined by the 2006 World Health Organization criteria. Spearman correlations were used to investigate the agreement between markers of hyperglycemia, and a receiver operating characteristic (ROC) curve was calculated to evaluate the use of A1C to identify newly diagnosed diabetes. RESULTS: In the total population, the correlations between fasting plasma glucose and A1C and between 2-h postload plasma glucose and A1C were 0.46 and 0.33, respectively. In patients with known diabetes, these correlations were 0.71 and 0.79. An A1C level of > or =5.8%, representing 12% of the population, had the highest combination of sensitivity (72%) and specificity (91%) for identifying newly diagnosed diabetes. This cutoff point would identify 72% of the patients with newly diagnosed diabetes and include 30% of the individuals with intermediate hyperglycemia. CONCLUSIONS: In patients with known diabetes, correlations between glucose and A1C are strong; however, moderate correlations were found in the general population. In addition, based on the diagnostic properties of A1C defined by ROC curve analysis, the advantage of A1C compared with OGTT for the diagnosis of diabetes is limited. PMID- 19808929 TI - Many Americans have pre-diabetes and should be considered for metformin therapy. AB - OBJECTIVE: To determine the proportion of the American population who would merit metformin treatment, according to recent American Diabetes Association (ADA) consensus panel recommendations to prevent or delay the development of diabetes. RESEARCH DESIGN AND METHODS: Risk factors were evaluated in 1,581 Screening for Impaired Glucose Tolerance (SIGT), 2,014 Third National Health and Nutrition Examination Survey (NHANES III), and 1,111 National Health and Nutrition Examination Survey 2005-2006 (NHANES 2005-2006) subjects, who were non-Hispanic white and black, without known diabetes. Criteria for consideration of metformin included the presence of both impaired fasting glucose (IFG) and impaired glucose tolerance (IGT), with > or =1 additional diabetes risk factor: age <60 years, BMI > or =35 kg/m(2), family history of diabetes, elevated triglycerides, reduced HDL cholesterol, hypertension, or A1C >6.0%. RESULTS: Isolated IFG, isolated IGT, and IFG and IGT were found in 18.0, 7.2, and 8.2% of SIGT; 22.3, 6.4, and 9.4% of NHANES III; and 21.8, 5.0, and 9.0% of NHANES 2005-2006 subjects, respectively. In SIGT, NHANES III, and NHANES 2005-2006, criteria for metformin consideration were met in 99, 96, and 96% of those with IFG and IGT; 31, 29, and 28% of all those with IFG; and 53, 57, and 62% of all those with IGT (8.1, 9.1, and 8.7% of all subjects), respectively. CONCLUSIONS: More than 96% of individuals with both IFG and IGT are likely to meet ADA consensus criteria for consideration of metformin. Because >28% of all those with IFG met the criteria, providers should perform oral glucose tolerance tests to find concomitant IGT in all patients with IFG. To the extent that our findings are representative of the U.S. population, approximately 1 in 12 adults has a combination of pre-diabetes and risk factors that may justify consideration of metformin treatment for diabetes prevention. PMID- 19808930 TI - Specific relation between abdominal obesity and early-phase hyperglycemia is modulated by hepatic insulin resistance in healthy older women. AB - OBJECTIVE: To describe the impact of abdominal obesity and hepatic insulin resistance on phase-specific glycemic responses in older women. RESEARCH DESIGN AND METHODS: We studied 23 healthy older women (60-88 years old). Abdominal obesity was defined by an abdominal circumference > or =95 cm. Plasma glucose and insulin were measured in response to a 3-h oral glucose tolerance test. Insulin suppression of hepatic glucose production was determined using in vivo clamp techniques. RESULTS: Despite identical prevailing insulin concentrations, glucose excursions 30 min postchallenge (but not later) were greater in women with abdominal obesity than in those without (162 +/- 19 vs. 132 +/- 16 mg/dl; P < 0.01). There was a strong correlation between hepatic glucose production suppression under low-dose insulin infusion and early-phase glucose excursions from the oral glucose tolerance test (r = -0.83; P < 0.001) in women with abdominal obesity, but not in women without (r = 0.44; P < 0.11). CONCLUSIONS: Abdominal obesity relates specifically to early-phase hyperglycemia via hepatic insulin resistance, even in healthy older women. PMID- 19808931 TI - Insights into the nature and consequences of our variable genome. PMID- 19808932 TI - Sequencing genomes: from individuals to populations. AB - The whole genome sequences of Jim Watson and Craig Venter are early examples of personalized genomics, which promises to change how we approach healthcare in the future. Before personal sequencing can have practical medical benefits, however, and before it should be advocated for implementation at the population-scale, there needs to be a better understanding of which genetic variants influence which traits and how their effects are modified by epigenetic factors. Nonetheless, for forging links between DNA sequence and phenotype, efforts to sequence the genomes of individuals need to continue; this includes sequencing sub-populations for association studies which analyse the difference in sequence between disease affected and unaffected individuals. Such studies can only be applied on a large enough scale to be effective if the massive strides in sequencing technology that have recently occurred also continue. PMID- 19808933 TI - MEPE/OF45 protects cells from DNA damage induced killing via stabilizing CHK1. AB - Matrix extracellular phosphoglycoprotein/osteoblast factor 45 (MEPE/OF45) was cloned in 2000 with functions related to bone metabolism. We identified MEPE/OF45 for the first time as a new co-factor of CHK1 in mammalian cells to protect cells from DNA damage induced killing. We demonstrate here that MEPE/OF45 directly interacts with CHK1. Knocking down MEPE/OF45 decreases CHK1 levels and sensitizes the cells to DNA damage inducers such as ionizing radiation (IR) or camptothicin (CPT)-induced killing. Over-expressing wild-type MEPE/OF45, but not the mutant MEPE/OF45 (depleted the key domain to interact with CHK1) increases CHK1 levels in the cells and increases the resistance of the cells to IR or CPT. MEPE/OF45, interacting with CHK1, increases CHK1 half-life and decreases CHK1 degradation through the ubiquitine-mediated pathway. In addition, the interaction of MEPE/OF45 with CHK1 decreases CHK1 levels in the ubiquitin E3 ligases (Cul1 and Cul4A) complex, which suggests that MEPE/OF45 competes with the ubiquitin E3 ligases binding to CHK1 and thus decreases CHK1 from ubiquitin-mediated proteolysis. These findings reveal an important role of MEPE/OF45 in protecting cells from DNA damage induced killing through stabilizing CHK1, which would provide MEPE/OF45 as a new target for sensitizing tumor cells to radiotherapy or chemotherapy. PMID- 19808934 TI - Specificity of LTR DNA recognition by a peptide mimicking the HIV-1 integrase {alpha}4 helix. AB - HIV-1 integrase integrates retroviral DNA through 3'-processing and strand transfer reactions in the presence of a divalent cation (Mg(2+) or Mn(2+)). The alpha4 helix exposed at the catalytic core surface is essential to the specific recognition of viral DNA. To define group determinants of recognition, we used a model composed of a peptide analogue of the alpha4 helix, oligonucleotides mimicking processed and unprocessed U5 LTR end and 5 mM Mg(2+). Circular dichroism, fluorescence and NMR experiments confirmed the implication of the alpha4 helix polar/charged face in specific and non-specific bindings to LTR ends. The specific binding requires unprocessed LTR ends-i.e. an unaltered 3' processing site CA downward arrowGT3'-and is reinforced by Mg(2+) (K(d) decreases from 2 to 0.8 nM). The latter likely interacts with the ApG and GpT3' steps of the 3'-processing site. With deletion of GT3', only persists non-specific binding (K(d) of 100 microM). Proton chemical shift deviations showed that specific binding need conserved amino acids in the alpha4 helix and conserved nucleotide bases and backbone groups at LTR ends. We suggest a conserved recognition mechanism based on both direct and indirect readout and which is subject to evolutionary pressure. PMID- 19808935 TI - PMRD: plant microRNA database. AB - MicroRNAs (miRNA) are approximately 21 nucleotide-long non-coding small RNAs, which function as post-transcriptional regulators in eukaryotes. miRNAs play essential roles in regulating plant growth and development. In recent years, research into the mechanism and consequences of miRNA action has made great progress. With whole genome sequence available in such plants as Arabidopsis thaliana, Oryza sativa, Populus trichocarpa, Glycine max, etc., it is desirable to develop a plant miRNA database through the integration of large amounts of information about publicly deposited miRNA data. The plant miRNA database (PMRD) integrates available plant miRNA data deposited in public databases, gleaned from the recent literature, and data generated in-house. This database contains sequence information, secondary structure, target genes, expression profiles and a genome browser. In total, there are 8433 miRNAs collected from 121 plant species in PMRD, including model plants and major crops such as Arabidopsis, rice, wheat, soybean, maize, sorghum, barley, etc. For Arabidopsis, rice, poplar, soybean, cotton, medicago and maize, we included the possible target genes for each miRNA with a predicted interaction site in the database. Furthermore, we provided miRNA expression profiles in the PMRD, including our local rice oxidative stress related microarray data (LC Sciences miRPlants_10.1) and the recently published microarray data for poplar, Arabidopsis, tomato, maize and rice. The PMRD database was constructed by open source technology utilizing a user-friendly web interface, and multiple search tools. The PMRD is freely available at http://bioinformatics.cau.edu.cn/PMRD. We expect PMRD to be a useful tool for scientists in the miRNA field in order to study the function of miRNAs and their target genes, especially in model plants and major crops. PMID- 19808936 TI - DNA cleavage and methylation specificity of the single polypeptide restriction modification enzyme LlaGI. AB - LlaGI is a single polypeptide restriction-modification enzyme encoded on the naturally-occurring plasmid pEW104 isolated from Lactococcus lactis ssp. cremoris W10. Bioinformatics analysis suggests that the enzyme contains domains characteristic of an mrr endonuclease, a superfamily 2 DNA helicase and a gamma family adenine methyltransferase. LlaGI was expressed and purified from a recombinant clone and its properties characterised. An asymmetric recognition sequence was identified, 5'-CTnGAyG-3' (where n is A, G, C or T and y is C or T). Methylation of the recognition site occurred on only one strand (the non degenerate dA residue of 5'-CrTCnAG-3' being methylated at the N6 position). Double strand DNA breaks at distant, random sites were only observed when two head-to-head oriented, unmethylated copies of the site were present; single sites or pairs in tail-to-tail or head-to-tail repeat only supported a DNA nicking activity. dsDNA nuclease activity was dependent upon the presence of ATP or dATP. Our results are consistent with a directional long-range communication mechanism that is necessitated by the partial site methylation. In the accompanying manuscript [Smith et al. (2009) The single polypeptide restriction-modification enzyme LlaGI is a self-contained molecular motor that translocates DNA loops], we demonstrate that this communication is via 1-dimensional DNA loop translocation. On the basis of this data and that in the third accompanying manuscript [Smith et al. (2009) An Mrr-family nuclease motif in the single polypeptide restriction modification enzyme LlaGI], we propose that LlaGI is the prototype of a new sub classification of Restriction-Modification enzymes, named Type I SP (for Single Polypeptide). PMID- 19808937 TI - Expanded RNA-binding activities of mammalian Argonaute 2. AB - Mammalian Argonaute 2 (Ago2) protein associates with microRNAs (miRNAs) or small interfering RNAs (siRNAs) forming RNA-induced silencing complexes (RISCs/miRNPs). In the present work, we characterize the RNA-binding and nucleolytic activity of recombinant mouse Ago2. Our studies show that recombinant mouse Ago2 binds efficiently to miRNAs forming active RISC. Surprisingly, we find that recombinant mouse Ago2 forms active RISC using pre-miRNAs or long unstructured single stranded RNAs as guides. Furthermore, we demonstrate that, in vivo, endogenous human Ago2 binds directly to pre-miRNAs independently of Dicer, and that Ago2:pre miRNA complexes are found both in the cytoplasm and in the nucleus of human cells. PMID- 19808938 TI - New tools at the Candida Genome Database: biochemical pathways and full-text literature search. AB - The Candida Genome Database (CGD, http://www.candidagenome.org/) provides online access to genomic sequence data and manually curated functional information about genes and proteins of the human pathogen Candida albicans. Herein, we describe two recently added features, Candida Biochemical Pathways and the Textpresso full text literature search tool. The Biochemical Pathways tool provides visualization of metabolic pathways and analysis tools that facilitate interpretation of experimental data, including results of large-scale experiments, in the context of Candida metabolism. Textpresso for Candida allows searching through the full text of Candida-specific literature, including clinical and epidemiological studies. PMID- 19808939 TI - Human defensins and LL-37 in mucosal immunity. AB - Defensins are widespread in nature and have activity against a broad range of pathogens. Defensins have direct antimicrobial effects and also modulate innate and adaptive immune responses. We consider the role of human defensins and the cathelicidin LL-37 in defense of respiratory, gastrointestinal, and genitourinary tracts and the oral cavity, skin, and eye. Human beta-defensins (hBDs) and human defensins 5 and 6 (HD5 and -6) are involved most obviously in mucosal responses, as they are produced principally by epithelial cells. Human alpha-defensins 1-4 (or HNPs 1-4) are produced principally by neutrophils recruited to the mucosa. Understanding the biology of defensins and LL-37 is the beginning to clarify the pathophysiology of mucosal inflammatory and infectious diseases (e.g., Crohn's disease, atopic dermatitis, lung or urinary infections). Challenges for these studies are the redundancy of innate defense mechanisms and the presence and interactions of many innate defense proteins in mucosal secretions. PMID- 19808940 TI - The long arm of expectancies: adolescent alcohol expectancies predict adult alcohol use. AB - AIMS: Alcohol expectancies are strong concurrent predictors of alcohol use and problems, but the current study addressed their unique power to predict from adolescence to midlife. METHOD: Long-term longitudinal data from the national British Cohort Study 1970 (N = 2146, 59.8% female) were used to predict alcohol use and misuse in the mid-30s by alcohol expectancies reported in adolescence. RESULTS: Cohort members with more positive alcohol expectancies at age 16 reported greater alcohol quantity concurrently, increases in alcohol quantity relative to their peers between ages 16 and 35, and a higher likelihood of lifetime and previous year alcohol misuse at age 35, independent of gender, social class in family of origin, age of alcohol use onset, adolescent delinquent behavior and age 16 exam scores. CONCLUSIONS: Alcohol expectancies were strong proximal predictors of alcohol use and predicted relative change in alcohol use and misuse across two decades into middle adulthood. PMID- 19808941 TI - Relationship between alcohol consumption and active Helicobacter pylori infection. AB - BACKGROUND: Helicobacter pylori (H. pylori) is a cause of chronic gastritis and maybe responsible for functional dyspepsia in a subset of patients. Many risk factors, such as alcohol consumption and smoking, may contribute to the colonization and infection of H. pylori in humans. However, studies on the relationship between H. pylori infection and drinking or smoking have produced conflicting results. OBJECTIVE: The aim of this study was to examine whether consumption of alcohol or smoking is associated with active H. pylori infection in functional dyspepsia patients. METHODS: H. pylori infection was confirmed by CLOtest and histology on at least two biopsies. Active chronic gastritis was diagnosed using the updated Sydney system. In addition to gender and age, information on drinking and smoking habits was collected using a standard questionnaire. Functional dyspepsia was diagnosed according to the Rome II diagnostic criteria. RESULTS: H. pylori infection was positive in 27.3% of the 139 functional dyspepsia patients. Both age and gender were not significantly associated with H. pylori infection. A multiple logistic model found that alcohol consumption (OR = 9.05, 95% CI: 1.05-77.98) and pathology (active gastritis) (OR = 595.39, 95% CI: 81.43-4353.33) were associated with H. pylori infection. Active gastritis was associated with alcohol consumption (OR = 2.89, 95% CI: 1.03-8.02), smoking (OR = 2.72, 95% CI: 1.22-6.05) and age (OR = 1.03, 95% CI: 1.01-1.06). CONCLUSIONS: In patients with functional dyspepsia, there is no significant association between active H. pylori infection and smoking. However, alcohol consumption appears to be associated with H. pylori infection. PMID- 19808942 TI - Towards DSM-V: exploring diagnostic thresholds for alcohol dependence and abuse. AB - AIMS: The expected release of the DSM-V in 2012 has renewed the longstanding debate around whether alcohol use disorders are best conceptualized as dimensional or categorical constructs. The current study aimed to validate the current diagnostic thresholds for alcohol dependence and abuse using epidemiological indicators including mental health, disability, psychological distress, functional impairment, service use, suicidality and early age of drinking onset. METHODS: Dichotomous variables were created to allocate a representative sample of Australian adult drinkers (n = 4920) above and below each possible threshold for both disorders. Regression analyses were conducted to assess group differences at each threshold for each epidemiological indicator. RESULTS: There was some albeit limited support for the current diagnostic threshold of three criteria for alcohol dependence and one criterion for abuse. A number of other cut-offs also showed consistent variation for both disorders. CONCLUSIONS: It is essential to define diagnostic thresholds in a systematic way. The current diagnostic thresholds for alcohol dependence and abuse are adequate but require further validation using a variety of methods and external indicators. Combining these disorders in some way may also prove useful as well as including other potential diagnostic criteria in future research. PMID- 19808943 TI - Methodological assessment of economic evaluations of alcohol treatment: what is missing? AB - AIM: The aim of this study is to review the methodology that has been adopted in previous economic evaluations of alcohol treatment and offer research recommendations with a view to enhancing the consistency and harmonization of economic evaluations in the alcohol field. METHODS: Published full economic evaluations of alcohol treatment were retrieved using a systematic search. The studies were analysed in terms of the identification, measurement and valuation methods used to assess the society-level consequences and the methods used to carry out the analysis of individual-level consequences and costs of the intervention. A taxonomy of alcohol-related consequences was developed and used as a framework for the methodology extraction. RESULTS: Twenty- seven studies were selected. Almost half of the studies did not include society-level consequences in their analysis. Some consequences of alcohol treatment at a societal level, such as the impact of treatment on health-related quality of life of family and friends of the drinker, have never been considered in the economic analysis. There was no agreement regarding the individual health consequences used in the evaluations. Measures capturing life years and morbidity have not been extensively used in the alcohol field. The level of reporting treatment costs on the reviewed studies is generally well detailed. CONCLUSION: The literature is still rather sparse in this area and further research is required to fulfil the gaps. If a common methodology is adopted in future economic evaluations of alcohol treatment, more stable cost-effectiveness estimates will be produced and informed decisions for resources allocation to alcohol treatments will be possible. PMID- 19808944 TI - Treating type 2 diabetes: incretin mimetics and enhancers. AB - As a consequence of excess abdominal adiposity and genetic predisposition, type 2 diabetes is a progressive disease, often diagnosed after metabolic dysfunction has taken hold of multiple organ systems. Insulin deficiency, insulin resistance and impaired glucose homeostasis resulting from beta-cell dysfunction characterize the disease. Current treatment goals are often unmet due to insufficient treatment modalities. Even when combined, these treatment modalities are frequently limited by safety, tolerability, weight gain, edema and gastrointestinal intolerance. Recently, new therapeutic classes have become available for treatment. This review will examine the new therapeutic classes of incretin mimetics and enhancers in the treatment of type 2 diabetes. PMID- 19808945 TI - OWL1 is a phytochrome a signaling component dedicated to the very low fluence response. PMID- 19808947 TI - Focal segmental glomerulosclerosis in idiopathic membranous glomerulonephritis: a clinico-pathological and stereological study. AB - BACKGROUND: The phenomenon of focal segmental glomerulosclerosis (FSGS) in idiopathic membranous glomerulonephritis (IMGN) has not been adequately studied. There is also a paucity of detailed glomerular morphometric and stereologic analyses data on renal biopsy in this association. METHODS: Twenty-three (23) patients with IMGN and superimposed FSGS were compared to 35 patients with IMGN alone with respect to the clinical and laboratory features, light microscopic findings and stereologic parameters (glomerular cross-sectional area and estimated glomerular volume). RESULTS: In the clinical parameters, patients with IMGN-FSGS had a significantly higher incidence of hypertension, raised serum creatinine and microscopic haematuria. The mean 24-h urinary protein excretion was higher in the group with IMGN-FSGS (7.4 +/- 1.36 g) as compared to IMGN alone (3.85 +/- 0.7 g, P < 0.001, Mann-Whitney test). On light microscopy, biopsies with IMGN-FSGS frequently had mesangial hypercellularity and more extensive tubulo-interstitial disease than those with IMGN alone. Stereological analysis showed that the non-sclerosed glomeruli in biopsies with IMGN-FSGS had a higher mean cross-sectional area (185466.7 +/- 32493.3 micro(2)) and higher estimated volume (855200 +/- 152640 micro(3)) as compared to glomeruli in cases with IMGN alone (76000 +/- 14719.2 micro(2) and 576666.7 +/- 131233.3 micro(3), respectively). CONCLUSION: The present study is probably the first systematic analysis of stereologic parameters in renal biopsies of IMGN with FSGS. Our results objectively demonstrate the glomerular enlargement in the non-sclerosed glomeruli in cases of IMGN with FSGS. This detection of enlarged glomeruli may serve to alert the renal pathologist to the possibility of coexisting FSGS, which is a poor prognostic factor in IMGN. PMID- 19808948 TI - Is progression of IgA nephropathy conditioned by genes regulating atherosclerotic damage? PMID- 19808946 TI - OWL1: an Arabidopsis J-domain protein involved in perception of very low light fluences. AB - To sense ambient light conditions in order to optimize their growth and development, plants employ a battery of photoreceptors responsive to light quality and quantity. Essential for the sensing of red and far-red (FR) light is the phytochrome family of photoreceptors. Among them, phytochrome A is special because it mediates responses to different light conditions, including both very low fluences (very low fluence response [VLFR]) and high irradiances (high irradiance response [HIR]). In contrast with the FR-HIR signaling pathway, in which several intermediates of the signaling pathway have been identified, specific components of the VLFR pathway remain unknown. Here, we describe owl1 (for orientation under very low fluences of light), a mutant that is specific for the VLFR, suggesting that VLFR and HIR pathways are genetically distinct, although some common mechanisms can be observed. OWL1 codes for a ubiquitous J domain protein essential for germination, cotyledon opening, hypocotyl elongation, and deviation of the direction of hypocotyl growth from the vertical under very low light conditions. Additionally, we observed a flowering phenotype suggesting a role for the VLFR during the whole life cycle of a plant. OWL1 interacts with the basic helix-loop-helix HFR1 (LONG HYPOCOTYL IN FAR-RED) transcription factor, previously characterized as a component of the FR-HIR pathway. Both proteins are involved in the agravitropic response under FR light. We propose a central function of OWL1 in the VLFR pathway, which is essential for plant survival under unfavorable light conditions. PMID- 19808949 TI - Evaluation of clinical dry weight assessment in haemodialysis patients using bioimpedance spectroscopy: a cross-sectional study. AB - BACKGROUND: Dry weight assessment (DWA) is essential to efficient therapy of haemodialysis (HD) patients. However, so far objective methods for DWA have not been applicable to daily routine. Thus, exact fluid management in HD remains difficult and is often based on clinical criteria. The aims of this study were (1) to objectively define pre- and post-dialytic ranges of extracellular volume in a large cohort of HD patients (in whom DWA had been defined according to clinical criteria), (2) to compare the hydration status between diabetic and non diabetic patients, and (3) to assess a patient subgroup that might benefit from correction of target weight. METHODS: We measured fluid overload (FO) prior to a mid-week HD session in 370 randomly selected HD patients (50% with diabetes) from five dialysis centres. A new bioimpedance spectroscopy (BIS) device that implies a validated body composition model was applied. This tool allows correct quantification of extracellular FO or - deficiency in comparison to a healthy reference population (normal range -1.1 to 1.1 L according to the 10th and 90th percentile of measurements). In addition, weight and blood pressure were recorded before and after treatment. RESULTS: Pre-dialytic FO ranged from -0.5 to 4 L and post-dialytic FO from -2.5 to 2 L (10th and 90th percentile of measurements), indicating that on average the hydration status of healthy subjects is considered as the optimal target weight in HD patients. Comparison of FO between diabetic and non-diabetic patients revealed no difference. Based on the consideration that an FO < -1.1 L before and >1.1 L after HD indicates inadequate DWA, we identified 98 (26%) patients who might benefit from correction of target body weight. CONCLUSION: BIS is an interesting, objective method to support clinical DWA. Further studies should be performed to investigate beneficial clinical effects of this approach. PMID- 19808950 TI - Pleiotropic effects of atorvastatin on monocytes in atherosclerotic patients. AB - The objective of this study was to investigate the gene expression signature of monocyte/macrophages and the pleiotropic effects of atorvastatin on monocytes in atherosclerotic patients. Forty patients with coronary heart diseases were randomly assigned to double-blind therapy with either 20 or 80 mg per day of atorvastatin. Follow-up visits occurred at weeks 6 and 12, including complete chemistry and lipid analyses and quantification of 14 target genes in monocytes. After 12 weeks of therapy, both groups gained beneficial alterations in lipid profiles. Both groups experienced significant reductions in gene expression of lipoprotein-associated phospholipase A2, CD13, leptin receptor, matrix metalloproteases-1, legumain, and prolyl oligopeptidase after 12 weeks of therapy. Only tumor protein 53 was increased in the atorvastatin 80-mg group. Moreover, nonsignificant interactions between dosage and duration of therapy were found. The pleiotropic effects of statins in atherosclerotic patients include increased expression of genes involved in apoptosis of monocyte/macrophage, inhibition of inflammatory responses, antioxidant properties, prevention of foam cell formation, and stabilization of atherosclerotic plaques. This property fuels potential clinical significance. PMID- 19808951 TI - A phase I study to characterize the safety, tolerability, and pharmacokinetics of topotecan at 4 mg/m2 administered weekly as a 30-minute intravenous infusion in patients with cancer. AB - Topotecan pharmacokinetics at higher infusion rates (4 mg/m2 over 30 minutes) have not been studied. The authors report a pharmacokinetics and safety study of this dose in advanced cancer patients. Sixteen patients were given a 4-mg/m2 topotecan infusion intravenously (IV) over 30 minutes weekly for 3 weeks, repeated every 28 days. Pharmacokinetics were determined after the first dose. Plasma concentrations of total topotecan were measured to derive CL, V(ss), C(max), t(max), t(1/2), AUC(0-t), and AUC(0-infinity). Plasma total topotecan concentrations decreased biexponentially, with a mean CL value of 20.6 L/h, V(ss) value of 101 L, and t(1/2) value of 5.0 h. Nine significant adverse events (all hematologic) were topotecan related. Grade 3 or less adverse events included anemia, thrombocytopenia, leukopenia, and fatigue. Pharmacokinetics of the 4 mg/m2 infusion of topotecan over 30 minutes are comparable to findings from studies of lower and higher doses. Toxicities are similar to previous reports. PMID- 19808952 TI - Effect of thyroid hormone on the activity of CYP3A enzyme in humans. AB - Thyroid hormones have been shown to reduce the activity and expression of cytochrome P450 (CYP) 3A4 in vitro. The influence of thyroid hormone on drug action via a CYP3A-dependent pathway has not been elucidated in humans. This is the first report showing the effect of thyroid hormone on CYP3A enzyme activity in humans. Ten healthy volunteers participate in this open-label study, in which the pharmacokinetics of midazolam and the urinary ratios of 6 beta hydroxycortisol/free cortisol before and after 2 weeks of oral administration of triiodothyronine were compared. Triiodothyronine administration significantly reduced the area under the concentration-time curve ratios for 1' hydroxymidazolam/midazolam from 0.36 to 0.25 (P < .05) and urinary ratios of 6 beta-hydroxycortisol/free cortisol from 6.92 to 5.88 (P < .05). These results strongly suggested that thyroid hormone reduced CYP3A activity in human and may influence the pharmacokinetics of concomitant CYP3A substrate drugs. PMID- 19808953 TI - Height, body mass index, and physical activity in relation to glioma risk. AB - Whether energy balance during early life and/or adulthood is related to glioma risk is unknown. We therefore investigated height, body mass index (BMI), and physical activity in relation to glioma risk in the prospective NIH-AARP Diet and Health Study. Participants completed a baseline questionnaire (sent in 1995-1996) inquiring about height, weight, and potential confounders. A second questionnaire (sent in 1996) inquired about physical activity during ages 15 to 18, 19 to 29, and 35 to 39 years and the past 10 years and body weight at ages 18, 35, and 50 years. During follow-up from 1995/1996 to 2003, we documented 480 cases of glioma among 499,437 respondents to the baseline questionnaire and 257 cases among 305,681 respondents to the second questionnaire. Glioma risk among tall persons (>or=1.90 m) was twice that of short persons [<1.60 m; multivariate relative risk (RR), 2.12; 95% confidence interval (95% CI), 1.18-3.81; P(trend) = 0.006]. Risk among participants who were obese (BMI 30.0-34.9 kg/m(2)) at age 18 years was nearly four times that of persons of normal weight (BMI 18.5-24.9 kg/m(2)) at age 18 years (RR, 3.74; 95% CI, 2.03-6.90; P(trend) = 0.003); 11 cases were obese at age 18 years. Risk among participants who were active during ages 15 to 18 years was 36% lower than that of persons who were inactive during ages 15 to 18 years (RR, 0.64; 95% CI, 0.44-0.93; P(trend) = 0.02). BMI and physical activity after age 18 years were unrelated to glioma risk. Adult height, BMI during adolescence, and physical activity during adolescence were each associated with glioma risk, supporting a role for early-life energy balance in glioma carcinogenesis. PMID- 19808954 TI - Enhanced leukemia cell detection using a novel magnetic needle and nanoparticles. AB - Acute leukemia is a hematopoietic malignancy for which the accurate measurement of minimal residual disease is critical to determining prognosis and treatment. Although bone marrow aspiration and light microscopy remain the current standard of care for detecting residual disease, these approaches cannot reliably discriminate less than 5% lymphoblast cells. To improve the detection of leukemia cells in the marrow, we developed a novel apparatus that utilizes antibodies conjugated to superparamagnetic iron oxide nanoparticles (SPION) and directed against the acute leukemia antigen CD34, coupled with a "magnetic needle" biopsy. Leukemia cell lines expressing high or minimal CD34 were incubated with anti-CD34 conjugated SPIONs. Three separate approaches including microscopy, superconducting quantum interference device magnetometry, and in vitro magnetic needle extraction were then used to assess cell sampling. We found that CD34 conjugated nanoparticles preferentially bind high CD34-expressing cell lines. Furthermore, the magnetic needle enabled identification of both cell line and patient leukemia cells diluted into normal blood at concentrations below those normally found in remission marrow samples. Finally, the magnetic needle enhanced the percentage of lymphoblasts detectable by light microscopy by 10-fold in samples of fresh bone marrow aspirate approximating minimal residual disease. These data suggest that bone marrow biopsy using antigen-targeted magnetic nanoparticles and a magnetic needle for the evaluation of minimal residual disease in CD34-positive acute leukemias can significantly enhance sensitivity compared with the current standard of care. PMID- 19808955 TI - Frontiers of Biomedical Imaging Science 2009: workshop report and research opportunities. PMID- 19808956 TI - Tpl2 is a key mediator of arsenite-induced signal transduction. AB - Arsenite is a well-known human carcinogen that especially targets skin. The tumor progression locus 2 (Tpl2) gene encodes a serine/threonine protein kinase that is overexpressed in various cancer cells. However, the relevance of Tpl2 in arsenite induced carcinogenesis and the underlying mechanisms remain to be explored. We show that arsenite increased Tpl2 kinase activity and its phosphorylation in mouse epidermal JB6 P+ cells in a dose- and time-dependent manner. Exposure to arsenite resulted in a marked induction of cyclooxygenase-2 (COX-2) and prostaglandin E(2) (PGE(2)), important mediators of inflammation and tumor promotion. Treatment with a Tpl2 kinase inhibitor or Tpl2 short hairpin RNA suppressed COX-2 expression and PGE(2) production induced by arsenite treatment, suggesting that Tpl2 is critical in arsenite-induced carcinogenesis. We also found that arsenite-induced phosphorylation of extracellular signal-regulated kinases (ERK) or c-Jun NH(2)-terminal kinases (JNK) was markedly suppressed by Tpl2 kinase inhibitor or Tpl2 short hairpin RNA. Inhibition of arsenite-induced ERK or JNK signaling using a pharmacologic inhibitor of ERK or JNK substantially blocked COX-2 expression. Furthermore, inhibition of Tpl2 reduced the arsenite induced promoter activity of NF-kappaB and activator protein-1 (AP-1), indicating that NF-kappaB and AP-1 are downstream transducers of arsenite-triggered Tpl2. Our results show that Tpl2 plays a key role in arsenite-induced COX-2 expression and PGE(2) production and further elucidate the role of Tpl2 in arsenite signals that activate ERK/JNK and NF-kappaB/AP-1 in JB6 P+ cells. PMID- 19808957 TI - Tumor antigen-specific FOXP3+ CD4 T cells identified in human metastatic melanoma: peptide vaccination results in selective expansion of Th1-like counterparts. AB - We have previously shown that vaccination of HLA-A2 metastatic melanoma patients with the analogue Melan-A(26-35(A27L)) peptide emulsified in a mineral oil induces ex vivo detectable specific CD8 T cells. These are further enhanced when a TLR9 agonist is codelivered in the same vaccine formulation. Interestingly, the same peptide can be efficiently recognized by HLA-DQ6-restricted CD4 T cells. We used HLA-DQ6 multimers to assess the specific CD4 T-cell response in both healthy individuals and melanoma patients. We report that the majority of melanoma patients carry high frequencies of naturally circulating HLA-DQ6-restricted Melan A-specific CD4 T cells, a high proportion of which express FOXP3 and proliferate poorly in response to the cognate peptide. Upon vaccination, the relative frequency of multimer+ CD4 T cells did not change significantly. In contrast, we found a marked shift to FOXP3-negative CD4 T cells, accompanied by robust CD4 T cell proliferation upon in vitro stimulation with cognate peptide. A concomitant reduction in TCR diversity was also observed. This is the first report on direct ex vivo identification of antigen-specific FOXP3+ T cells by multimer labeling in cancer patients and on the direct assessment of the impact of peptide vaccination on immunoregulatory T cells. PMID- 19808958 TI - The cytoskeleton protein filamin-A is required for an efficient recombinational DNA double strand break repair. AB - The human actin-binding protein filamin-A (also known as ABP-280) cross-links actin into a dynamic three-dimensional structure. It interacts with >45 proteins of diverse functions, serving as the scaffold in various signaling networks. BRCA2 is a protein that regulates RAD51-dependent recombinational repair of DNA double strand breaks (DSB). Proximate to the COOH terminus of the BRCA2 protein, a conserved and DNA binding domain (BRCA2-DBD) interacts with filamin-A and BCCIP. In this study, we sought to test the hypothesis that filamin-A influences homologous recombinational repair of DSB and the maintenance of genomic stability. We used three pairs of cell lines with normal and reduced filamin-A expression, including breast cancer and melanoma cells. We found that lack or reduction of filamin-A sensitizes cells to ionizing radiation, slows the removal of DNA damage-induced gammaH2AX nuclear foci, reduces RAD51 nuclear focus formation and recruitment to chromatin in response to irradiation, and results in a 2-fold reduction of homologous recombinational repair of DSB. Furthermore, filamin-A-deficient cells have increased frequencies of micronucleus formation after irradiation. Our data illustrate the importance of the cytoskeleton structure in supporting the homologous recombinational DNA repair machinery and genome integrity, and further implicate a potential of filamin-A as a marker for prognosis in DNA damage-based cancer therapy. PMID- 19808959 TI - Sox11 prevents tumorigenesis of glioma-initiating cells by inducing neuronal differentiation. AB - Recent findings have shown that malignant tumors contain cancer-initiating cells (CIC), which self-renew and are tumorigenic. However, CICs have not been characterized properly due to lack of specific markers. We recently established a mouse glioma cell line, NSCL61, by overexpressing an oncogenic HRas(L61) in p53 deficient neural stem cells. Using limiting dilution assays, we show that only 2 of 24 NSCL61 clones retained their tumorigenicity in vivo, although the others also expressed oncogenic HRas(L61) and could proliferate in culture. A comparison of the gene expression profiles of tumorigenic and nontumorigenic clones showed that the tumorigenic clones had lost Sox11 expression. We show that overexpression of sox11 prevented tumorigenesis of NSCL61s by inducing their neuronal differentiation accompanied with decreased levels of plagl1. We also show that overexpression of plagl1 abolished neuronal commitment of nontumorigenic cells and induced them to become tumorigenic. Moreover, we show that human glioma-initiating cells lost sox11 expression, and overexpression of sox11 prevented their tumorigenesis in vivo. Together with the clinical evidence showing that downregulation of sox11 mRNA correlates with a significant decrease in survival, these findings suggest that Sox11 prevents gliomagenesis by blocking the expression of oncogenic plagl1. PMID- 19808960 TI - Apolipoprotein E/C1 locus variants modify renal cell carcinoma risk. AB - Lipid peroxidation is considered a unifying mechanistic pathway through which known risk factors induce renal cell carcinoma (RCC). We hypothesized that genes selected a priori for their role in lipid peroxidation would modify cancer risk. We genotyped 635 single nucleotide polymorphisms (SNP) in 38candidate genes in 777 Caucasian RCC cases and 1,035 controls enrolled in a large European case control study. Top candidate SNPs were confirmed among 718 Caucasian cases and 615 controls in a second study in the United States. Two of the three SNPs (rs8106822 and rs405509) that replicated in the U.S. study were within a regulatory region of the APOE promoter. The OR for rs8106822 A>G variant was 1.22(AG) and 1.41(GG) (P(trend) = 0.01) in the European study, 1.05(AG) and 1.51(GG) (P(trend) = 0.03) in the U.S. study, and 1.15(AG) and 1.44(GG) (P(trend) = 0.001) among 1,485 cases and 1,639 controls combined. The rs405509 G>T variant was associated with risk in the European (OR, 0.87(TG); OR, 0.71(TT); P(trend) = 0.02), the U.S. (OR, 0.68(TG); OR, 0.71(TT); P(trend) = 0.02), and both studies combined (OR(TG), 0.79; OR(TT), 0.71; P(trend) = 0.001), as was the G-G haplotype (r(2) = 0.64; P= 4.7 x 10(-4)). This association is biologically plausible as SNP rs405509 was shown to modify protein binding and transcriptional activity of the APOE protein in vitro and is in linkage disequilibrium with key known variants defining the e2, e3, and e4 alleles that modify risk of atherosclerosis, Alzheimer's disease risk, and progression to AIDS. In two large case-control studies, our findings further define a functional region of interest at the APOE locus that increases RCC susceptibility. PMID- 19808961 TI - Revving the Throttle on an oncogene: CDK8 takes the driver seat. AB - The Wnt/beta-catenin pathway plays an important role in initiation in most, if not all, colon cancers. Prior work has provided important insights into the regulation of beta-catenin stability in the cytoplasm; however, relatively little is known about the mechanism by which beta-catenin activates gene transcription in the nucleus. Using genetic approaches, studies in human colon cancers and Drosophila have identified CDK8 as a colon cancer oncogene that regulates beta catenin transcriptional activity. These convergent observations provide new insights into the regulation of nuclear beta-catenin activity and identify a novel therapeutic target for beta-catenin-driven malignancies. PMID- 19808962 TI - Differences in tumor regulatory T-cell localization and activation status impact patient outcome. AB - The presence of regulatory T cells (Treg) has been described in a large panel of solid tumors. However, their impact on tumor progression differs according to the tumor type analyzed. We recently obtained evidence in breast carcinoma that Treg localized within lymphoid aggregates, but not in the tumor bed, have a negative impact on patients' survival. Moreover, we showed selective Treg recruitment through CCR4/CCL22 in the lymphoid aggregates upon contact with dendritic cells (DC), where they became strongly and selectively activated (ICOS(high)) and block conventional T-cell response. Here, we discuss the meaning and potential implication of these novel findings. PMID- 19808963 TI - Structural basis for the binding of the anticancer compound 6-(7-nitro-2,1,3 benzoxadiazol-4-ylthio)hexanol to human glutathione s-transferases. AB - Glutathione S-transferases (GST) constitute a superfamily of enzymes with diversified functions including detoxification from xenobiotics. In many human cancers, Pi class GST (GSTP1-1) is overexpressed and contributes to multidrug resistance by conjugating chemotherapeutics. In addition, GSTP1-1 displays antiapoptotic activity by interacting with c-Jun NH(2)-terminal kinase, a key regulator of apoptosis. Therefore, GSTP1-1 is considered a promising target for pharmaceutical treatment. Recently, a potent inhibitor of GSTs, 6-(7-nitro-2,1,3 benzoxadiazol-4-ylthio)hexanol (NBDHEX), was identified and tested on several tumor cell lines demonstrating high antiproliferative activity. To establish the structural basis of NBDHEX activity, we determined the crystal structure of NBDHEX bound to either GSTP1-1 or GSTM2-2 (mu class). NBDHEX in both cases binds to the H-site but occupies different positions. Furthermore, the compound is covalently attached to the GSH sulfur in the GSTM2-2 crystal, forming a sigma complex, although it is bound but not conjugated in the GSTP1-1 crystal. Several differences in the H-sites of the two isozymes determine the higher affinity of NBDHEX for GSTM2-2 with respect to GSTP1-1. One such difference is the presence of Ile(104) in GSTP1-1 close to the bound NBDHEX, whereas the corresponding position is occupied by an alanine in GSTM2-2. Mutation of Ile(104) into valine is a frequent GSTP1-1 polymorphism and we show here that the Ile(104)Val and Ile(104)Ala variants display a 4-fold higher affinity for the compound. Remarkably, the GSTP1-1/Ile(104)Ala structure in complex with NBDHEX shows a considerable shift of the compound inside the H-site. These data might be useful for the development of new anticancer compounds. PMID- 19808964 TI - A novel PTEN-dependent link to ubiquitination controls FLIPS stability and TRAIL sensitivity in glioblastoma multiforme. AB - Phosphatase and tensin homologue (PTEN) loss and activation of the Akt-mammalian target of rapamycin (mTOR) pathway increases mRNA translation, increases levels of the antiapoptotic protein FLIP(S), and confers resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in glioblastoma multiforme (GBM). In PTEN-deficient GBM cells, however, the FLIP(S) protein also exhibited a longer half-life than in PTEN mutant GBM cells, and this longer half-life correlated with decreased FLIP(S) polyubiquitination. FLIP(S) half-life in PTEN mutant GBM cells was reduced by exposure to an Akt inhibitor, but not to rapamycin, suggesting the existence of a previously undescribed, mTOR independent linkage between PTEN and the ubiquitin-dependent control of protein stability. Total levels of the candidate FLIP(S) E3 ubiquitin ligase atrophin interacting protein 4 (AIP4) were comparable in PTEN wild-type (WT) and PTEN mutant GBM cells, although in PTEN-deficient cells, AIP4 was maintained in a stable polyubiquitinated state that was less able to associate with FLIP(S) or with the FLIP(S)-containing death inducing signal complex. Small interfering RNA mediated suppression of AIP4 levels in PTEN WT cells decreased FLIP(S) ubiquitination, prolonged FLIP(S) half-life, and increased TRAIL resistance. Similarly, the Akt activation that was previously shown to increase TRAIL resistance did not alter AIP4 levels, but increased AIP4 ubiquitination, increased FLIP(S) steady-state levels, and suppressed FLIP(S) ubiquitination. These results define the PTEN-Akt-AIP4 pathway as a key regulator of FLIP(S) ubiquitination, FLIP(S) stability, and TRAIL sensitivity and also define a novel link between PTEN and the ubiquitin-mediated control of protein stability. PMID- 19808965 TI - A modified sleeping beauty transposon system that can be used to model a wide variety of human cancers in mice. AB - Recent advances in cancer therapeutics stress the need for a better understanding of the molecular mechanisms driving tumor formation. This can be accomplished by obtaining a more complete description of the genes that contribute to cancer. We previously described an approach using the Sleeping Beauty (SB) transposon system to model hematopoietic malignancies in mice. Here, we describe modifications of the SB system that provide additional flexibility in generating mouse models of cancer. First, we describe a Cre-inducible SBase allele, RosaSBase(LsL), that allows the restriction of transposon mutagenesis to a specific tissue of interest. This allele was used to generate a model of germinal center B-cell lymphoma by activating SBase expression with an Aid-Cre allele. In a second approach, a novel transposon was generated, T2/Onc3, in which the CMV enhancer/chicken beta-actin promoter drives oncogene expression. When combined with ubiquitous SBase expression, the T2/Onc3 transposon produced nearly 200 independent tumors of more than 20 different types in a cohort of 62 mice. Analysis of transposon insertion sites identified novel candidate genes, including Zmiz1 and Rian, involved in squamous cell carcinoma and hepatocellular carcinoma, respectively. These novel alleles provide additional tools for the SB system and provide some insight into how this mutagenesis system can be manipulated to model cancer in mice. PMID- 19808966 TI - Aldehyde dehydrogenase-expressing colon stem cells contribute to tumorigenesis in the transition from colitis to cancer. AB - Patients with chronic ulcerative colitis are at increased risk of developing colorectal cancer. Although current hypotheses suggest that sporadic colorectal cancer is due to inability to control cancer stem cells, the cancer stem cell hypothesis has not yet been validated in colitis-associated cancer. Furthermore, the identification of the colitis to cancer transition is challenging. We recently showed that epithelial cells with the increased expression of aldehyde dehydrogenase in sporadic colon cancer correlate closely with tumor-initiating ability. We sought to determine whether ALDH can be used as a marker to isolate tumor-initiating populations from patients with chronic ulcerative colitis. We used fluorescence-activated cell sorting to identify precursor colon cancer stem cells from colitis patients and report both their transition to cancerous stem cells in xenografting studies as well as their ability to generate spheres in vitro. Similar to sporadic colon cancer, these colitis-derived tumors were capable of propagation as sphere cultures. However, unlike the origins of sporadic colon cancer, the primary colitic tissues did not express any histologic evidence of dysplasia. To elucidate a potential mechanism for our findings, we compared the stroma of these different environments and determined that at least one paracrine factor is up-regulated in the inflammatory and malignant stroma compared with resting, normal stroma. These data link colitis and cancer identifying potential tumor-initiating cells from colitic patients, suggesting that sphere and/or xenograft formation will be useful to survey colitic patients at risk of developing cancer. PMID- 19808967 TI - Runt-related transcription factor RUNX3 is a target of MDM2-mediated ubiquitination. AB - The p14(ARF)-MDM2-p53 pathway constitutes an effective mechanism for protecting cells from oncogenic stimuli such as activated Ras and Myc. Importantly, Ras activation induces p14(ARF) and often occurs earlier than p53 inactivation during cancer development. Here, we show that RUNX3, a tumor suppressor in various tumors including stomach, bladder, colon, and lung, is stabilized by Ras activation through the p14(ARF)-MDM2 signaling pathway. RUNX3 directly binds MDM2 through its Runt-related DNA-binding domain. MDM2 blocks RUNX3 transcriptional activity by interacting with RUNX3 through an acidic domain adjacent to the p53 binding domain of MDM2 and ubiquitinates RUNX3 on key lysine residues to mediate nuclear export and proteasomal degradation. Our data indicate that the lineage specific tumor suppressor RUNX3 and the ubiquitous p53 protein are both principal responders of the p14(ARF)-MDM2 cell surveillance pathway that prevents pathologic consequences of abnormal oncogene activation. PMID- 19808968 TI - Increased expression of androgen receptor sensitizes prostate cancer cells to low levels of androgens. AB - Androgen receptor (AR) is known to be overexpressed in castration-resistant prostate cancer. To interrogate the functional significance of the AR level, we established two LNCaP cell sublines expressing in a stable fashion two to four times (LNCaP-ARmo) and four to six times (LNCaP-ARhi) higher level of AR than the parental cell line expressing the empty vector (LNCaP-pcDNA3.1). LNCaP-ARhi cell line grew faster than the control line in low concentrations, especially in 1 nmol/L 5alpha-dihydrotestosterone (DHT). Microarray-based transcript profiling and subsequent unsupervised hierarchical clustering showed that LNCaP-ARhi cells clustered together with VCaP cells, containing endogenous AR gene amplification and overexpression, indicating the central role of AR in the overall regulation of gene expression in prostate cancer cells. Two hundred forty genes showed >2 fold changes on DHT treatment in LNCaP-ARhi at 4 h time point, whereas only 164 and 52 showed changes in LNCaP-ARmo and LNCaP-pcDNA3.1, respectively. Many androgen-regulated genes were upregulated in LNCaP-ARhi at 10-fold lower concentration of DHT than in control cells. DHT (1 nmol/L) increased expression of several cell cycle-associated genes in LNCaP-ARhi cells. ChIP-on-chip assay revealed the presence of chromatin binding sites for AR within +/-200 kb of most of these genes. The growth of LNCaP-ARhi cells was also highly sensitive to cyclin-dependent kinase inhibitor, roscovitine, at 1nmol/L DHT. In conclusion, our results show that overexpression of AR sensitizes castration-resistant prostate cancer cells to the low levels of androgens. The activity of AR signaling pathway is regulated by the levels of both ligand and the receptor. PMID- 19808969 TI - EBV-induced human CD8+ NKT cells suppress tumorigenesis by EBV-associated malignancies. AB - The underlying mechanism of the protective and suppressive role of NKT cells in human tumor immunosurveillance remains to be fully elucidated. We show that the frequencies of CD8(+) NKT cells in patients with EBV-associated Hodgkin's lymphoma or nasopharyngeal carcinoma are significantly lower than those in healthy EBV carriers. These CD8(+) NKT cells in tumor patients are also functionally impaired. In human-thymus-severe combined immunodeficient (hu-thym SCID) chimeras, EBV challenge efficiently promotes the generation of IFN-gamma biased CD8(+) NKT cells. These cells are strongly cytotoxic, drive syngeneic T cells into a Th1 bias, and enhance T-cell cytotoxicity to EBV-associated tumor cells. Interleukin-4-biased CD4(+) NKT cells are predominately generated in unchallenged chimeras. These cells are noncytotoxic, drive syngeneic T cells into a Th2 bias, and do not affect T-cell cytotoxicity. In humanized xenogeneic tumor transplanted hu-thym-SCID chimeras, adoptive transfer with EBV-induced CD8(+) NKT cells significantly suppresses tumorigenesis by EBV-associated malignancies. EBV induced CD8(+) NKT cells are necessary and sufficient to enhance the T-cell immunity to EBV-associated malignancies in the hu-thym-SCID chimeras. CD4(+) NKT cells are synergetic with CD8(+) NKT cells, leading to a more pronounced T-cell antitumor response in the chimeras cotransferred with CD4(+) and CD8(+) NKT cells. Thus, immune reconstitution with EBV-induced CD8(+) NKT cells could be a useful strategy in management of EBV-associated malignancies. PMID- 19808970 TI - Phosphorylation and degradation of MdmX is inhibited by Wip1 phosphatase in the DNA damage response. AB - MdmX and Mdm2 regulate p53 tumor suppressor functions by controlling p53 transcriptional activity and/or stability in cells exposed to DNA damage. Accumulating evidence indicates that ATM-mediated phosphorylation and degradation of Mdm2 and MdmX may be the initial driving force that induces p53 activity during the early phase of the DNA damage response. We have recently determined that a novel protein phosphatase, Wip1 (or PPM1D), contributes to p53 regulation by dephosphorylating Mdm2 to close the p53 activation loop initiated by the ATM/ATR kinases. In the present study, we determine that Wip1 directly dephosphorylates MdmX at the ATM-targeted Ser403 and indirectly suppresses phosphorylation of MdmX at Ser342 and Ser367. Wip1 inhibits the DNA damage induced ubiquitination and degradation of MdmX, leading to the stabilization of MdmX and reduction of p53 activities. Our data suggest that Wip1 is an important component in the ATM-p53-MdmX regulatory loop. PMID- 19808971 TI - Azacytidine inhibits RNA methylation at DNMT2 target sites in human cancer cell lines. AB - The cytosine analogues azacytidine and decitabine are currently being developed as drugs for epigenetic cancer therapy. Although various studies have shown that both drugs are effective in inhibiting DNA methylation, it has also become clear that their mode of action is not limited to DNA demethylation. Because azacytidine is a ribonucleoside, the primary target of this drug may be cellular RNA rather than DNA. We have now analyzed the possibility that azacytidine inhibits the RNA methyltransferase DNMT2. We found that DNMT2 is variably expressed in human cancer cell lines. RNA bisulfite sequencing showed that azacytidine, but not decitabine, inhibits cytosine 38 methylation of tRNA(Asp), a major substrate of DNMT2. Azacytidine caused a substantially stronger effect than decitabine on the metabolic rate of all the cancer cell lines tested, consistent with an effect of this drug on RNA metabolism. Of note, drug-induced loss of RNA methylation seemed specific for DNMT2 target sites because we did not observe any significant demethylation at sites known to be methylated by other RNA methyltransferases. Our results uncover a novel and quantifiable drug activity of azacytidine and raise the possibility that tRNA hypomethylation might contribute to patient responses. PMID- 19808972 TI - A novel experimental heme oxygenase-1-targeted therapy for hormone-refractory prostate cancer. AB - Heme oxygenase-1 (HO-1), a member of the heat shock protein family, plays a key role as a sensor and regulator of oxidative stress. Herein, we identify HO-1 as a biomarker and potential therapeutic target for advanced prostate cancer (PCA). Immunohistochemical analysis of prostate tissue using a progression tissue microarray from patients with localized PCA and across several stages of disease progression revealed a significant elevation of HO-1 expression in cancer epithelial cells, but not in surrounding stromal cells, from hormone-refractory PCA (HRPCA) compared with hormone-responsive PCA and benign tissue. Silencing the ho-1 gene in HRPCA cells decreased the HO-1 activity, oxidative stress, and activation of the mitogen-activated protein kinase-extracellular signal-regulated kinase/p38 kinase. This coincided with reduced cell proliferation, cell survival, and cell invasion in vitro, as well as inhibition of prostate tumor growth and lymph node and lung metastases in vivo. The effect of ho-1 silencing on these oncogenic features was mimicked by exposure of cells to a novel selective small molecule HO-1 inhibitor referred to as OB-24. OB-24 selectively inhibited HO-1 activity in PCA cells, which correlated with a reduction of protein carbonylation and reactive oxygen species formation. Moreover, OB-24 significantly inhibited cell proliferation in vitro and tumor growth and lymph node/lung metastases in vivo. A potent synergistic activity was observed when OB-24 was combined with Taxol. Together, these results establish HO-1 as a potential therapeutic target for advanced PCA. PMID- 19808973 TI - Inhibition of tumor cell growth, invasion, and metastasis by EXEL-2880 (XL880, GSK1363089), a novel inhibitor of HGF and VEGF receptor tyrosine kinases. AB - The Met receptor tyrosine kinase and its ligand, hepatocyte growth factor (HGF), are overexpressed and/or activated in a wide variety of human malignancies. Vascular endothelial growth factor (VEGF) receptors are expressed on the surface of vascular endothelial cells and cooperate with Met to induce tumor invasion and vascularization. EXEL-2880 (XL880, GSK1363089) is a small-molecule kinase inhibitor that targets members of the HGF and VEGF receptor tyrosine kinase families, with additional inhibitory activity toward KIT, Flt-3, platelet-derived growth factor receptor beta, and Tie-2. Binding of EXEL-2880 to Met and VEGF receptor 2 (KDR) is characterized by a very slow off-rate, consistent with X-ray crystallographic data showing that the inhibitor is deeply bound in the Met kinase active site cleft. EXEL-2880 inhibits cellular HGF-induced Met phosphorylation and VEGF-induced extracellular signal-regulated kinase phosphorylation and prevents both HGF-induced responses of tumor cells and HGF/VEGF-induced responses of endothelial cells. In addition, EXEL-2880 prevents anchorage-independent proliferation of tumor cells under both normoxic and hypoxic conditions. In vivo, these effects produce significant dose-dependent inhibition of tumor burden in an experimental model of lung metastasis. Collectively, these data indicate that EXEL-2880 may prevent tumor growth through a direct effect on tumor cell proliferation and by inhibition of invasion and angiogenesis mediated by HGF and VEGF receptors. PMID- 19808974 TI - ARTEMIS nuclease facilitates apoptotic chromatin cleavage. AB - One hallmark of apoptosis is DNA degradation that first appears as high molecular weight fragments followed by extensive internucleosomal fragmentation. During apoptosis, the DNA-dependent protein kinase (DNA-PK) is activated. DNA-PK is involved in the repair of DNA double-strand breaks (DSB) and its catalytic subunit is associated with the nuclease ARTEMIS. Here, we report that, on initiation of apoptosis in human cells by agents causing DNA DSB or by staurosporine or other agents, ARTEMIS binds to apoptotic chromatin together with DNA-PK and other DSB repair proteins. ARTEMIS recruitment to chromatin showed a time and dose dependency. It required DNA-PK protein kinase activity and was blocked by antagonizing the onset of apoptosis with a pan-caspase inhibitor or on overexpression of the antiapoptotic BCL2 protein. In the absence of ARTEMIS, no defect in caspase-3, poly(ADP-ribose) polymerase-1, and XRCC4 cleavage or in H2AX phosphorylation was observed and DNA-PK catalytic subunit was still phosphorylated on S2056 in response to staurosporine. However, DNA fragmentation including high molecular weight fragmentation was delayed in ARTEMIS-deficient cells compared with cells expressing ARTEMIS. In addition, ARTEMIS enhanced the kinetics of MLL gene cleavage at a breakage cluster breakpoint that is frequently translocated in acute or therapy-related leukemias. These results show a facilitating role for ARTEMIS at least in early, site-specific chromosome breakage during apoptosis. PMID- 19808975 TI - A peptide fragment of azurin induces a p53-mediated cell cycle arrest in human breast cancer cells. AB - We report that amino acids 50 to 77 of azurin (p28) preferentially enter the human breast cancer cell lines MCF-7, ZR-75-1, and T47D through a caveolin mediated pathway. Although p28 enters p53 wild-type MCF-7 and the isogenic p53 dominant-negative MDD2 breast cancer cell lines, p28 only induces a G(2)-M-phase cell cycle arrest and apoptosis in MCF-7 cells. p28 exerts its antiproliferative activity by reducing proteasomal degradation of p53 through formation of a p28:p53 complex within a hydrophobic DNA-binding domain (amino acids 80-276), increasing p53 levels and DNA-binding activity. Subsequent elevation of the cyclin-dependent kinase inhibitors p21 and p27 reduces cyclin-dependent kinase 2 and cyclin A levels in a time-dependent manner in MCF-7 cells but not in MDD2 cells. These results suggest that p28 and similar peptides that significantly reduce proteasomal degradation of p53 by a MDM2-independent pathway(s) may provide a unique series of cytostatic and cytotoxic (apoptotic) chemotherapeutic agents. PMID- 19808976 TI - Apomab, a fully human agonistic antibody to DR5, exhibits potent antitumor activity against primary and metastatic breast cancer. AB - Apomab, a fully human agonistic DR5 monoclonal antibody, triggers apoptosis through activation of the extrinsic apoptotic signaling pathway. In this study, we assessed the cytotoxic effect of Apomab in vitro and evaluated its antitumor activity in murine models of breast cancer development and progression. MDA-MB 231-TXSA breast cancer cells were transplanted into the mammary fat pad or directly into the tibial marrow cavity of nude mice. Apomab was administered early, postcancer cell transplantation, or after tumors progressed to an advanced stage. Tumor burden was monitored progressively using bioluminescence imaging, and the development of breast cancer-induced osteolysis was measured using microcomputed tomography. In vitro, Apomab treatment induced apoptosis in a panel of breast cancer cell lines but was without effect on normal human primary osteoblasts, fibroblasts, or mammary epithelial cells. In vivo, Apomab exerted remarkable tumor suppressive activity leading to complete regression of well advanced mammary tumors. All animals transplanted with breast cancer cells directly into their tibiae developed large osteolytic lesions that eroded the cortical bone. In contrast, treatment with Apomab following an early treatment protocol inhibited both intraosseous and extraosseous tumor growth and prevented breast cancer-induced osteolysis. In the delayed treatment protocol, Apomab treatment resulted in the complete regression of advanced tibial tumors with progressive restoration of both trabecular and cortical bone leading to full resolution of osteolytic lesions. Apomab represents a potent immunotherapeutic agent with strong activity against the development and progression of breast cancer and should be evaluated in patients with primary and metastatic disease. PMID- 19808977 TI - In vivo effects of targeting CD79b with antibodies and antibody-drug conjugates. AB - Antibodies directed against B cells are in use for the treatment of non-Hodgkin's lymphoma and autoimmune disorders. The B-cell-restricted surface antigen CD79b, a signaling component of the B-cell receptor, has been shown as a promising antibody target in mouse efficacy models of systemic lupus erythematosus. Anti CD79b antibody-drug conjugates (ADC), cytotoxic drugs linked through specialized chemical linkers to antibodies, are effective in mouse xenograft models of non Hodgkin's lymphoma. We were interested in evaluating the systemic effects of anti CD79b antibodies and ADCs in normal animals as a step toward the development of these molecules as therapeutics. As we were unable to identify any cell surface binding anti-human CD79b antibodies that were cross-reactive to other species, we developed an antibody to cynomolgus monkey (Macaca fascicularis) CD79b (anti cyCD79b). The anti-cynomolgus antibody, anti-cyCD79b (10D10), and the maytansine (tubulin inhibitor)-conjugated ADC, anti-cyCD79b (10D10)-MCC-DM1, were administered to cynomolgus monkeys at approximately 30 mg/kg (6,000 microg DM1/m(2)) for two doses 3 weeks apart. Anti-cyCD79b and anti-cyCD79b-MCC-DM1 resulted in peripheral blood B-cell depletion of approximately 65% and approximately 94%, respectively. In addition, anti-cyCD79b-MCC-DM1 resulted in near-complete absence of splenic germinal centers, an observation supporting an effect on dividing B cells. Both molecules were well tolerated, with minimal findings for the antibody and findings for the ADC limited to the lymphoid and hematopoietic systems, liver, and peripheral nerves. These preclinical data suggest that targeting CD79b with antibodies or ADCs may provide safe and effective therapies for B-cell malignancies and autoimmune diseases. PMID- 19808978 TI - Quantification of endothelial cell-targeted anti-Bcl-2 therapy and its suppression of tumor growth and vascularization. AB - Proapoptotic and antiapoptotic proteins in the Bcl family are key regulators of programmed cell death. It is the interaction between these molecules that determines cellular response to apoptotic signals, making them attractive targets for therapeutic intervention. In recent experiments designed to study tumor angiogenesis, Bcl-2 upregulation in endothelial cells was shown to be a critical mediator of vascular development. In this article, we develop a mathematical model that explicitly incorporates the response of endothelial cells to variations in proapoptotic and antiapoptotic proteins in the Bcl family, as well as the administration of specific antiangiogenic therapies targeted against Bcl 2. The model is validated by comparing its predictions to in vitro experimental data that reports microvessel density prior to and following the administration of 0.05 to 5.0 micromol/L of BL193, a promising small molecule inhibitor of Bcl 2. Numerical simulations of in vivo treatment of tumors predict the existence of a threshold for the amount of therapy required for successful treatment and quantify how this threshold varies with the stage of tumor growth. Furthermore, the model shows how rapidly the least effective dosage of BL193 decreases if an even moderately better inhibitor of Bcl-2 is used and predicts that increasing cell wall permeability of endothelial cells to BL193 does not significantly affect this threshold. A critical challenge of experimental therapeutics for cancer is to decide which drugs are the best candidates for clinical trials. These results underscore the potential of mathematical modeling to guide the development of novel antiangiogenic therapies and to direct drug design. PMID- 19808979 TI - Triptolide is an inhibitor of RNA polymerase I and II-dependent transcription leading predominantly to down-regulation of short-lived mRNA. AB - Triptolide, a natural product extracted from the Chinese plant Tripterygium wilfordii, possesses antitumor properties. Despite numerous reports showing the proapoptotic capacity and the inhibition of NF-kappaB-mediated transcription by triptolide, the identity of its cellular target is still unknown. To clarify its mechanism of action, we further investigated the effect of triptolide on RNA synthesis in the human non-small cell lung cancer cell line A549. Triptolide inhibited both total RNA and mRNA de novo synthesis, with the primary action being on the latter pool. We used 44K human pan-genomic DNA microarrays and identified the genes primarily affected by a short treatment with triptolide. Among the modulated genes, up to 98% are down-regulated, encompassing a large array of oncogenes including transcription factors and cell cycle regulators. We next observed that triptolide induced a rapid depletion of RPB1, the RNA polymerase II main subunit that is considered a hallmark of a transcription elongation blockage. However, we also show that triptolide does not directly interact with the RNA polymerase II complex nor does it damage DNA. We thus conclude that triptolide is an original pharmacologic inhibitor of RNA polymerase activity, affecting indirectly the transcription machinery, leading to a rapid depletion of short-lived mRNA, including transcription factors, cell cycle regulators such as CDC25A, and the oncogenes MYC and Src. Overall, the data shed light on the effect of triptolide on transcription, along with its novel potential applications in cancers, including acute myeloid leukemia, which is in part driven by the aforementioned oncogenic factors. PMID- 19808980 TI - BH3-only proteins Mcl-1 and Bim as well as endonuclease G are targeted in spongistatin 1-induced apoptosis in breast cancer cells. AB - Spongistatin 1, a marine experimental substance with chemotherapeutic potential, induces apoptosis and inhibits clonogenic survival of MCF-7 cells. Regarding the apoptotic signaling pathways of spongistatin 1, we present two major facts. Firstly, spongistatin 1-induced cell death, mainly caspase-independent, involves the proapoptotic proteins apoptosis-inducing factor and endonuclease G. Both proteins translocate from mitochondria to the nucleus and contribute to spongistatin 1-mediated apoptosis as shown via gene silencing. Secondly, spongistatin 1 acts as a tubulin depolymerizing agent and is able to free the proapoptotic Bcl-2 family member Bim from its sequestration both by the microtubular complex and by the antiapoptotic protein Mcl-1. Silencing of Bim by small interfering RNA leads to a diminished translocation of apoptosis-inducing factor and endonuclease G to the nucleus and subsequently reduces apoptosis rate. Thus, we identified Bim as an important factor upstream of mitochondria executing a central role in the caspase-independent apoptotic signaling pathway induced by spongistatin 1. Taken together, spongistatin 1 is both a valuable tool for the characterization of apoptotic pathways and a promising experimental anticancer drug. PMID- 19808981 TI - Improved ATM kinase inhibitor KU-60019 radiosensitizes glioma cells, compromises insulin, AKT and ERK prosurvival signaling, and inhibits migration and invasion. AB - Ataxia telangiectasia (A-T) mutated (ATM) is critical for cell cycle checkpoints and DNA repair. Thus, specific small molecule inhibitors targeting ATM could perhaps be developed into efficient radiosensitizers. Recently, a specific inhibitor of the ATM kinase, KU-55933, was shown to radiosensitize human cancer cells. Herein, we report on an improved analogue of KU-55933 (KU-60019) with K(i) and IC(50) values half of those of KU-55933. KU-60019 is 10-fold more effective than KU-55933 at blocking radiation-induced phosphorylation of key ATM targets in human glioma cells. As expected, KU-60019 is a highly effective radiosensitizer of human glioma cells. A-T fibroblasts were not radiosensitized by KU-60019, strongly suggesting that the ATM kinase is specifically targeted. Furthermore, KU 60019 reduced basal S473 AKT phosphorylation, suggesting that the ATM kinase might regulate a protein phosphatase acting on AKT. In line with this finding, the effect of KU-60019 on AKT phosphorylation was countered by low levels of okadaic acid, a phosphatase inhibitor, and A-T cells were impaired in S473 AKT phosphorylation in response to radiation and insulin and unresponsive to KU 60019. We also show that KU-60019 inhibits glioma cell migration and invasion in vitro, suggesting that glioma growth and motility might be controlled by ATM via AKT. Inhibitors of MEK and AKT did not further radiosensitize cells treated with KU-60019, supporting the idea that KU-60019 interferes with prosurvival signaling separate from its radiosensitizing properties. Altogether, KU-60019 inhibits the DNA damage response, reduces AKT phosphorylation and prosurvival signaling, inhibits migration and invasion, and effectively radiosensitizes human glioma cells. PMID- 19808982 TI - Nitro-chloromethylbenzindolines: hypoxia-activated prodrugs of potent adenine N3 DNA minor groove alkylators. AB - Hypoxia represents an important therapeutic target in tumors because of the resistance of hypoxic cells to radiotherapy and chemotherapy and because it is more severe in many tumors than in normal tissues. Here, we describe a class of prodrugs, nitro-chloromethylindolines, which undergo hypoxia-selective activation by endogenous nitroreductases in tumor cells to form the corresponding amino compounds. The latter are chemically related to the cyclopropylindoline antitumor antibiotics and they share the same properties of sequence-selective DNA minor groove alkylation and high cytotoxic potency. Of three alkylating subunits investigated, the chloromethylbenzindoline (CBI) structure provided the most favorable prodrug properties: aerobic cytotoxic potency of the amines was approximately 90- to 3,000-fold higher than the corresponding nitro compounds, and the nitro compounds showed air/anoxia potency differentials of up to 300 fold. Selective alkylation of adenine N3 in calf thymus DNA by an amino-CBI was shown by characterization of the thermal depurination product; the same adduct was shown in hypoxic RIF-1 cells exposed to the corresponding nitro-CBI prodrug under hypoxic (but not oxic) conditions. The amino metabolite generated from a nitro-CBI by cells expressing Escherichia coli nfsB nitroreductase in multicellular layer cultures was shown to elicit bystander killing of surrounding cells. Nitro-CBI prodrugs were >500-fold less toxic to mice than amino-CBIs by i.p. administration and provided selective killing of hypoxic cells in RIF-1 tumors (although only at maximally tolerated doses). Nitro-CBIs are novel lead hypoxia-activated prodrugs that represent the first examples of hypoxia-selective generation of potent DNA minor groove alkylating agents. PMID- 19808984 TI - BJSM reviews: A-Z of supplements: dietary supplements, sports nutrition foods and ergogenic aids for health and performance Part 1. PMID- 19808985 TI - Mutation analysis of the ASPM gene in 18 Pakistani families with autosomal recessive primary microcephaly. AB - Autosomal recessive primary microcephaly (MCPH) is a rare neurological disorder, in which the patients exhibit reduced occipital frontal head circumference (>3 standard deviations) and mild-to-severe mental retardation. Autosomal recessive primary microcephaly is genetically heterogeneous and 7 loci have been reported to date. Mutations in ASPM (abnormal spindle-like, microcephaly associated) gene are the most common cause of autosomal recessive primary microcephaly in the majority of the reported families. In the current investigation, we have located and studied 21 families with autosomal recessive primary microcephaly. Genotyping using polymorphic microsatellite markers linked to 7 autosomal recessive primary microcephaly loci revealed linkage of 18 families to the MCPH5 locus. Sequence analysis of the ASPM gene in 18 linked families detected 2 novel nonsense mutations (c.2101C>T/p.Q701X; c.9492T>G/p.Y3164X) in 2 families and 2 novel deletion mutations (c.6686delGAAA/p.R2229TfsX9; c.77delG/p.G26AfsX41) in 2 other families. Three previously described mutations (c.3978G>A/p.W1326X; c.1260delTCAAGTC/p.S420SfsX32; c.9159delA/p.K3053NfsX4) were also detected in 11 families. These identified mutations extended the body of evidence implicating the ASPM gene in the pathogenesis of human hereditary primary microcephaly. PMID- 19808986 TI - Major and minor arterial malformations in patients with cutaneous vascular abnormalities. AB - The association of persistent embryonic arteries and the absence of 1 carotid or vertebral arteries with facial or neck hemangioma or vascular malformation have been frequently described. The abnormalities can involve major or minor vessels. Of 22 patients of our series with this neurocutaneous syndrome, 20 had the origin of both anterior cerebral arteries from the same internal carotid artery. Thirteen patients showed absence or hypoplasia of 1 carotid artery and 10 of 1 vertebral artery; 10 showed persistence of the trigeminal artery; 3 had persistent proatlantal artery; 6 showed the absence of the posterior communicating artery; and 4 had hypoplastic posterior cerebral artery. Other less frequent abnormalities were found in 7 patients. Intellectual level of most patients was either borderline or below normal. Abnormalities in the vascularization and perfusion of the frontal lobes may contribute to the borderline or lower mental level of these patients. PMID- 19808987 TI - Early testing for Huntington disease in children: pros and cons. AB - We report 2 young children who are examples of the consequences of premature testing for Huntington disease. Premature testing of a child or fetus carries complex medical and psychological issues to both the child and the family that need to be considered and explored more than in an adult with Huntington disease. We suggest that a child at risk for juvenile Huntington disease not be tested until symptoms are progressive and consistent with the disease and all other mimickers are excluded. When testing is indicated, a multidisciplinary approach is essential to educate the family about the risks and benefits of testing and improve their coping skills when the final diagnosis is made. PMID- 19808988 TI - Outcome measures used in studies of botulinum toxin in childhood cerebral palsy: a systematic review. AB - This literature review uses the International Classification of Functioning, Disability and Health to describe the quality and variety of the studies of botulinum toxin in children with cerebral palsy since 2001. Articles were identified via electronic query and then reviewed for strength of evidence and classification of outcome measures. The distribution of levels of evidence for the 63 articles was I (n = 8), II (n = 12), III (n = 21), and IV (n = 22). One or more measurements were used in the International Classification of Functioning, Disability and Health domains of body structure and function (47 papers), activity (47 papers), participation (12 papers), and environmental/personal factors (10 papers). A total of 67% of all outcomes demonstrated a significant difference (I 49%, II 39%, III 74%, IV 88%). This review illustrates that few studies provide a high level of evidence and that outcomes focus on arenas such as spasticity or range of motion rather than activity or participation domains such as walking. PMID- 19808989 TI - A novel missense mutation in LIS1 in a child with subcortical band heterotopia and pachygyria inherited from his mildly affected mother with somatic mosaicism. AB - Mutations in the LIS1 gene result in isolated lissencephaly or subcortical band heterotopia. We report a 5-year-old male who presented with seizures and global developmental delay. Magnetic resonance imaging (MRI) demonstrated posteriorly predominant pachygyria and subcortical band heterotopia. His mother had a history of epilepsy, with onset in her teenage years. Her MRI revealed no abnormalities. Sequence analysis of the LIS1 gene identified a novel p.H389Y mutation in exon 11 (c.1165C>T). The child's mother was found to have the identical mutation as her son, with the signal intensity of the mutant allele being much lower than the normal allele, suggesting somatic mosaicism. This patient is one of only a few reported with a missense mutation in LIS1 associated with subcortical band heterotopia, and this is the first report of a mosaic individual having an affected child. PMID- 19808990 TI - Long-term prognosis of pediatric patients with relapsing acute disseminated encephalomyelitis. AB - Although long-term follow-up data are available for cases with acute disseminated encephalomyelitis, the findings range widely because of the lack of consistent definitions. Using the International Pediatric Multiple Sclerosis Study Group definitions strictly, we determined the long-term prognosis of children with acute disseminated encephalomyelitis, especially concerning relapsing cases. In our cohort of 86 children who presented with a first event of inflammatory demyelinating disease of central nervous system, 33 patients (38%) met the Study Group criteria for acute disseminated encephalomyelitis of which 9 patients had relapses. The mean follow-up duration was 12.8 years for relapsing cases and 9.2 years for all patients with acute disseminated encephalomyelitis. The risk of developing relapses is 27% but the risk of developing multiple sclerosis from acute disseminated encephalomyelitis is low at 6%. All relapsing cases had a benign course on prolonged follow-up, in spite of multiple relapses in the first 3 years. PMID- 19808991 TI - A case of secondary dystonia responding to levodopa. AB - The authors report a patient with dystonia secondary to bilateral lesions of the basal ganglia, who improved dramatically with levodopa. The patient presented at the age of 4 years with progressive dystonia of the lower extremities and right upper extremity. Magnetic resonance imaging (MRI) of the brain showed bilateral hyperintensities of the globus pallidus that remained stable over the years. Despite extensive investigations, the etiology of her basal ganglia lesions remained nebulous. The patient's dystonia responded to Trihexyphenidyl and to tetrabenazine, but these medications needed to be stopped because of side effects. At the age of 12 years, small doses of levodopa-carbidopa were tried and resulted in dramatic improvement of her dystonia. The authors believe that in the pediatric population with secondary dystonias other than Segawa disease, even though this has been reported only rarely to be effective, a therapeutic trial with levodopa should be considered in some instances. PMID- 19808992 TI - A delayed methadone encephalopathy: clinical and neuroradiological findings. AB - Several studies on opiates demonstrated that selected brain areas as cerebellum and limbic system have the greatest density of opioid receptors. Recently, few cases of severe cerebellitis following methadone poisoning have been reported in children. We present the case of a 30-month-old girl who developed a delayed encephalopathy after methadone intoxication. She was admitted to our emergency department in coma, and after naloxone infusion, she completely recovered. Five days after intoxication, she developed psychomotor agitation, slurred speech, abnormal movements, and ataxia despite a negative neuroimaging finding. A repeat magnetic resonance imaging (MRI) performed 19 days after the intoxication for persistent symptoms showed signal abnormalities in the temporomesial regions, basal ganglia, and substantia nigra. To our knowledge, this is the first report of these delayed MRI findings associated with synthetic opioid intoxication. PMID- 19808993 TI - Sociodemographic differences in diagnosis and treatment of pediatric headache. AB - The authors investigated the sociodemographic differences in receiving a headache diagnosis for pediatric health care visits using 2 nationally representative databases--the National Ambulatory Medical Care Survey and the National Hospital Ambulatory Medical Care Survey. For those visits involving a headache diagnosis, the authors explored 2 possible disparities in care--being diagnosed by a neurologist and being prescribed an evidence-based medication. A headache diagnosis was less common during visits for private insurance patients versus Medicaid patients. In addition, while a headache diagnosis was equally likely for visits by Caucasian American children versus African American children and children of other races, visits for the latter 2 groups were less likely to involve a headache diagnosis from a neurologist. Finally, only 37% of the headache visits involved a prescription of an evidence-based medication. The authors conclude that some sociodemographic disparities exist in pediatric headache care across the United States. PMID- 19808994 TI - Safe treatment of pulmonary hypertension with bosentan in a patient with moyamoya disease and cerebral ischemia. AB - A 7-year-old girl with Down syndrome and moderately severe pulmonary hypertension experienced a stroke while being treated with a calcium channel blocker. Angiography identified bilateral stenosis of the supraclinoid internal carotid arteries, stenosis or occlusion of the proximal anterior and middle cerebral arteries, and occlusion of the left posterior cerebral artery. She underwent surgery to enhance collateral blood flow to vulnerable areas of the brain. Her pulmonary hypertension therapy was changed to an oral endothelin receptor antagonist. She developed excellent collateral blood flow through external carotid arteries to each cerebral hemisphere and an improvement in blood flow through the right internal carotid artery. This case suggests that bosentan can be used safely in children with moyamoya disease. Additional studies are needed to determine whether endothelin receptor antagonists may influence the progression of moyamoya disease or the development of collateral cerebral blood flow following surgery. PMID- 19808996 TI - Health promotion practice: from vision to successful journal. PMID- 19808995 TI - Crofelemer, an antisecretory antidiarrheal proanthocyanidin oligomer extracted from Croton lechleri, targets two distinct intestinal chloride channels. AB - Crofelemer, a purified proanthocyanidin oligomer extracted from the bark latex of Croton lechleri, is in clinical trials for secretory diarrheas of various etiologies. We investigated the antisecretory mechanism of crofelemer by determining its effect on the major apical membrane transport and signaling processes involved in intestinal fluid transport. Using cell lines and measurement procedures to isolate the effects on individual membrane transport proteins, crofelemer at 50 microM had little or no effect on the activity of epithelial Na(+) or K(+) channels or on cAMP or calcium signaling. Crofelemer inhibited the cystic fibrosis transmembrane regulator (CFTR) Cl(-) channel with maximum inhibition of approximately 60% and an IC(50) approximately 7 microM. Crofelemer action at an extracellular site on CFTR produced voltage-independent block with stabilization of the channel closed state. Crofelemer did not affect the potency of glycine hydrazide or thiazolidinone CFTR inhibitors. Crofelemer action resisted washout, with <50% reversal of CFTR inhibition after 4 h. Crofelemer was also found to strongly inhibit the intestinal calcium-activated Cl(-) channel TMEM16A by a voltage-independent inhibition mechanism with maximum inhibition >90% and IC(50) approximately 6.5 microM. The dual inhibitory action of crofelemer on two structurally unrelated prosecretory intestinal Cl(-) channels may account for its intestinal antisecretory activity. PMID- 19808997 TI - Health promotion practice begins a new chapter. PMID- 19808998 TI - 50+ Ways to leave 'em learnin': tried and true training techniques. PMID- 19808999 TI - Advocacy evaluation: what it is and where to find out more about it. AB - Advocacy and policy change have become increasingly important strategies in health promotion efforts. As a result, advocacy evaluation is emerging as a necessary competency for health promotion practitioners. This article introduces the growing field of advocacy evaluation by describing its typical features, which include using prospective evaluation designs, emphasizing real-time data collection and use, monitoring and responding to changing environmental conditions, and assessing both individual and organizational capacity building. A number of freely available resources for learning more about advocacy evaluation's key concepts and methods are highlighted. PMID- 19809000 TI - Beyond the cathedral: building trust to engage the African American community in health promotion and disease prevention. AB - Effective efforts to eliminate health disparities must be grounded in strong community partnerships and trusting relationships between academic institutions and minority communities. However, there are often barriers to such efforts, including the frequent need to rely on time-limited funding mechanisms that take categorical approaches. This article provides an overview of health promotion and disease prevention projects implemented through the Community Outreach and Information Dissemination Core (COID) of the Center for Minority Health, within the Graduate School of Public Health at the University of Pittsburgh. The COID is one of five Cores that comprised the University of Pittsburgh's NIH Excellence in Partnerships for Community Outreach, and Research on Disparities in Health and Training (EXPORT Health) funded from 2002 to 2007 by the National Center on Minority Health and Health Disparities. Based in large part on the success of the community engagement activities, in 2007, the National Center on Minority Health and Health Disparities, National Institutes of Health, designated the CMH as a Research Center of Excellence on Minority Health Disparities. COID major initiatives included the Community Research Advisory Board, Health Disparity Working Groups, Health Advocates in Reach, Healthy Class of 2010, and the Healthy Black Family Project. Lessons learned may provide guidance to other academic institutions, community-based organizations, and health departments who seek to engage minority communities in changing social norms to support health promotion and disease prevention. PMID- 19809002 TI - Integrating cell phones and mobile technologies into public health practice: a social marketing perspective. AB - Mobile communications are being used for many purposes, from instant messaging (IM), mobile or microblogging (Twitter), social networking sites (Facebook, MySpace), e-mail to basic voicemail. A brief background on cell phone and mobile technology use in public health is reviewed. The focus of the article is framing the use of mobile technologies in public health from a social marketer's perspective--using the 4 Ps marketing mix as a guide. PMID- 19809003 TI - Marketing health educators to employers: survey findings, interpretations, and considerations for the profession. AB - In July 2007, a market research report was produced by Hezel Associates on behalf of five sponsoring health education profession member organizations and the National Commission for Health Education Credentialing. The purpose of the survey was to learn about current or potential employers' knowledge, attitudes, and behaviors toward health educators and the health education profession and their future hiring practices. This article presents the background leading up to the production of this report, the major findings of the survey of employers, recommendations from the market research group regarding core messages, and implications for the profession having discovered for the first time information about employers' understanding of professionally prepared health educators. The article discusses the umbrella and key messages that may be incorporated into a marketing plan and other recommendations by the firm that should assist health educators in marketing the profession. Furthermore, this article presents reactions by leaders in this field to these messages and recommendations and concludes with next steps in this project and a call for the overall need to market the profession of health education. PMID- 19809004 TI - Settings for health promotion: an analytic framework to guide intervention design and implementation. AB - Taking a settings approach to health promotion means addressing the contexts within which people live, work, and play and making these the object of inquiry and intervention as well as the needs and capacities of people to be found in different settings. This approach can increase the likelihood of success because it offers opportunities to situate practice in its context. Members of the setting can optimize interventions for specific contextual contingencies, target crucial factors in the organizational context influencing behavior, and render settings themselves more health promoting. A number of attempts have been made to systematize evidence regarding the effectiveness of interventions in different types of settings (e.g., school-based health promotion, community development). Few, if any, attempts have been made to systematically develop a template or framework for analyzing those features of settings that should influence intervention design and delivery. This article lays out the core elements of such a framework in the form of a nested series of questions to guide analysis. Furthermore, it offers advice on additional considerations that should be taken into account when operationalizing a settings approach in the field. PMID- 19809005 TI - Comparative pharmacodynamics of intermittent and prolonged infusions of piperacillin/tazobactam using Monte Carlo simulation and steady-state pharmacokinetic data from hospitalized patients. AB - With increasing antibiotic resistance in gram-negative pathogens, dosing strategies that optimize pharmacodynamic parameters of currently available antibiotics play an important role in treatment. The likelihood of success with piperacillin/tazobactam, a widely used broad-spectrum antibiotic, can be manipulated by increasing the amount of time that unbound drug concentrations remain above the pathogen's minimum inhibitory concentration (MIC). However, this success depends greatly on knowing the MIC value as well as accurately estimating the individual's pharmacokinetic parameters. Clinicians should carefully factor these variables into their decision-making process when considering prolonged infusion strategies with piperacillin/tazobactam. PMID- 19809006 TI - Bipolar disorder patient care opportunities: let's answer the call. AB - Bipolar disorder is a brain illness with complexities in its composition and treatment. Results from the National Comorbidity Survey Replication study estimate the lifetime prevalence of bipolar spectrum illnesses to be 4.5%. These patients were also reported to have their illnesses frequently treated suboptimally and to be at risk of suffering extensive disability. Pharmacists are in a key position to deliver important pharmacy care services to patients who have bipolar disorder and receive treatments in need of close monitoring. Described here are the results of this study and opportunities for pharmacists to support this important patient population. PMID- 19809007 TI - Bendamustine-associated hemolytic anemia. AB - OBJECTIVE: To report a case of probable bendamustine-related hemolytic anemia. CASE SUMMARY: A 64-year-old white female had recently received treatment with bendamustine for stage III follicular lymphoma. After her fourth cycle, she was admitted to an outside facility with severe right upper quadrant pain across her back and findings consistent with obstructive jaundice. She was found to have pancytopenia and elevations in total bilirubin, alkaline phosphatase, and transaminase levels. A bone marrow biopsy showed no evidence of lymphoma and presence of megakaryocytes on 2 occasions. Upon transfer to West Virginia University Hospitals, her haptoglobin was found to be undetectable, total bilirubin 10.3 mg/dL (unconjugated bilirubin 4.9 mg/dL), reticulocyte count 21.4% (reticulocyte index > or = 2%), alkaline phosphatase 1125 U/L, and lactate dehydrogenase 421 U/L. The peripheral smear showed evidence of spherocytes and very rare schistocytes. Based on these findings, the woman was diagnosed with hemolytic anemia secondary to bendamustine exposure. She was started on prednisone 1 mg/kg (60 mg) daily and, soon after, her platelets and hemoglobin stabilized. DISCUSSION: Drug-induced hemolytic anemia is an acquired or extrinsic process that results in antibody-mediated red blood cell destruction. The patient was not taking any medications commonly associated with hemolytic anemia; however, her laboratory test results were consistent with hemolytic anemia. Based on bendamustine's structural similarity to fludarabine and fludarabine's association with causing hemolytic anemia, we considered exposure to bendamustine to be the most likely contributory factor for her diagnosis. According to the Naranjo probability scale, a probable likelihood was reflected in bendamustine causing the hemolytic anemia. CONCLUSIONS: Continued monitoring of postmarketing data is necessary to correlate this occurrence of hemolytic anemia with bendamustine therapy. PMID- 19809008 TI - Use of duloxetine in pregnancy and lactation. AB - OBJECTIVE: To report a case of a woman who used duloxetine during pregnancy and breast-feeding. CASE SUMMARY: A 29-year-old woman was treated with duloxetine for depression during the second half of an uncomplicated gestation. She gave birth at term to a healthy female infant. A cord blood sample was obtained at birth. The mother continued the antidepressant while exclusively breast-feeding her infant. One month later, we collected blood and milk samples from the mother and a single blood sample from the infant. All samples were analyzed for the presence and concentrations of duloxetine. DISCUSSION: Duloxetine crosses the placenta at term and is excreted into breast milk. No evidence of developmental or other type of toxicity was observed in the infant at birth or during the first 32 days after birth. The published literature detailing human pregnancy experience with this antidepressant is limited to 11 cases in which women became pregnant while taking duloxetine. In 10 cases, the drug was discontinued when pregnancy was diagnosed and no outcome data were reported. In the eleventh case, an infant exposed to duloxetine 90 mg/day developed neonatal behavioral syndrome. One study examined the excretion of duloxetine into breast milk, but the mothers discontinued nursing for the study. In the present case, no adverse effects from exposure to the drug in milk were noted in the exclusively breast-fed infant. The possibility of functional/neurobehavioral deficits appearing later in life cannot be excluded because long-term follow-up has not been conducted in infants exposed to duloxetine in utero or during nursing. CONCLUSIONS: No developmental toxicity or other signs of toxicity were observed in an infant exposed to duloxetine during the second half of gestation and during breast-feeding in the first 32 days after birth. However, the possibility of functional/neurobehavioral deficits appearing later in life cannot be excluded. PMID- 19809009 TI - Comparative pharmacodynamics of intermittent and prolonged infusions of piperacillin/tazobactam using Monte Carlo simulations and steady-state pharmacokinetic data from hospitalized patients. AB - BACKGROUND: Prolonging the infusion of a beta-lactam antibiotic enhances the time in which unbound drug concentrations remain above the minimum inhibitory concentration (fT>MIC). OBJECTIVE: To compare the pharmacodynamics of several dosing regimens of piperacillin/tazobactam administered by intermittent and prolonged infusion using pharmacokinetic data from hospitalized patients. METHODS: Steady-state pharmacokinetic data were obtained from 13 patients who received piperacillin/tazobactam 4.5 g every 8 hours, infused over 4 hours. Monte Carlo simulations (10,000 pts.) were performed to calculate pharmacodynamic exposures at 50% fT>MIC for 4 intermittent-infusion regimens (3.375 g every 4 and 6 h, 4.5 g every 6 and 8 h) and 4 prolonged-infusion regimens (2.25 g, 3.375 g, 4.5 g, and 6.75 g every 8 h [4-h infusion]) of piperacillin/tazobactam using pharmacokinetic data for piperacillin. Cumulative fraction of response (CFR) was calculated using MIC data for 6 gram-negative pathogens (Meropenem Yearly Susceptibility Test Information Collection, 2004-2007), and probability of target attainment (PTA) was calculated at MICs ranging from 1 microg/mL to 64 microg/mL. RESULTS: The CFR for 3.375 g every 4 hours (intermittent infusion) and 3.375-4.5 g every 8 hours (prolonged infusion) greater than or equal to 90.3% for Escherichia coli, Serratia marcescens, and Citrobacter spp. Increasing the prolonged-infusion dose to 6.75 g improved the CFR to greater than 90% for Enterobacter spp. For every regimen evaluated, the CFR was less than 90% for Klebsiella pneumoniae and Pseudomonas aeruginosa. At an MIC of 16 microg/mL, PTA was greater than 90% for one intermittent-infusion regimen (3.375 g every 4 h) and 3 prolonged-infusion regimens (> or = 3.375 g every 8 h), but no regimen achieved a PTA greater than 90% at an MIC of 64 microg/mL. CONCLUSIONS: At doses greater than or equal to 3.375 g every 8 hours, 4-hour infusions of piperacillin/tazobactam achieved excellent target attainment with lower daily doses compared with standard regimens at MICs less than or equal to 16 microg/mL. PMID- 19809010 TI - Gastrointestinal safety of nonsteroidal antiinflammatory drugs and selective cyclooxygenase-2 inhibitors in patients on warfarin. AB - BACKGROUND: The interaction between warfarin and nonsteroidal antiinflammatory drugs (NSAIDs) is well known. However, warfarin and NSAIDs are still commonly prescribed together. Selective cyclooxygenase-2 (COX-2) inhibitors, a newer class of NSAID, offer potential advantages over the nonselective NSAIDs in patients treated with warfarin. OBJECTIVE: To study the rates of hospitalization for gastrointestinal (GI) bleeding events in 3 groups of patients: those taking warfarin only, those taking warfarin plus a nonselective NSAID, and those taking warfarin plus a selective COX-2 inhibitor. METHODS: This was a retrospective cohort analysis in a large nonprofit health maintenance organization. All warfarin users from January 1, 2000, to December 31, 2005, were eligible for inclusion in the study. Eligible patients were grouped by their exposure time to warfarin only, warfarin plus nonselective NSAIDs, or warfarin plus selective COX 2 inhibitor. The study endpoint was hospitalization for a GI bleed. Patients were matched using a propensity scoring methodology. A multivariate Cox proportional hazards model was used to estimate the hazard ratio for GI bleeding between patient cohorts, controlling for age, sex, baseline medical conditions, prior history of GI bleeding, and prescription drug use. RESULTS: The eligible population consisted of 35,548 patients undergoing 46,214 courses of warfarin therapy. The adjusted hazard ratio for hospital-associated GI bleeding in the warfarin plus nonselective NSAID group versus warfarin alone was 3.58 (95% CI 2.31 to 5.55; p < 0.01) and for warfarin plus selective COX-2 inhibitor versus warfarin alone was 1.71 (95% CI 0.60 to 4.84; p = 0.31). For nonselective NSAIDs plus warfarin versus selective COX-2 inhibitor plus warfarin, the adjusted hazard ratio was 3.69 (95% CI 1.42 to 9.60; p = 0.01). CONCLUSIONS: In general, nonselective NSAIDs and selective COX-2 inhibitors should be avoided in patients taking warfarin. In situations where patients require NSAIDs and cannot be managed using other therapies, our results suggest that selective COX-2 inhibitors are associated with fewer hospitalizations for GI bleeding. PMID- 19809011 TI - Safety and efficacy of quetiapine in bipolar depression. AB - OBJECTIVE: To review the clinical data investigating the efficacy and safety of quetiapine in bipolar depression. DATA SOURCES: Searches of MEDLINE and PubMed (1977-July 2009) were conducted using the key words quetiapine and bipolar depression. The references of literature found were cross-referenced. The pharmaceutical company that produces quetiapine was contacted to obtain the posters for the EMBOLDEN I and EMBOLDEN II trials. STUDY SELECTION AND DATA EXTRACTION: Only double-blind, placebo-controlled trials were included for review, as well as any subanalyses of the literature that matched this criterion. DATA SYNTHESIS: There was a total of 5 double-blind, placebo-controlled trials and 5 subanalyses reviewed. The results of these data demonstrated quetiapine's efficacy in the treatment of depressive phases of bipolar disorder, including statistically significant improvement in the Montgomery-Asberg Depression Rating Scale (MADRS). In the trials reviewed in this article, the change in MADRS scores ranged from -15.4 to -16.94 within the quetiapine groups, and from -10.26 to 11.93 in the placebo groups. There were also statistically significant improvements in the Hamilton Anxiety Rating Scale, the Short Form of the Quality of Life Enjoyment and Satisfaction Questionnaire, the Pittsburgh Sleep Quality Index, and the Sheehan Disability Scale. All of these trials had a duration of 8 weeks and therefore cannot be applied to the long-term use of quetiapine in bipolar depression. The most common adverse events were sedation, somnolence, and dry mouth. The overall dropout rates for the trials reviewed ranged from 24% to 47%. CONCLUSIONS: Based on the literature reviewed here, quetiapine appears to be a safe and efficacious short-term treatment option for bipolar depression. Patients with bipolar type I showed greater improvement on the MADRS than those with bipolar type II. Patients with a rapid-cycling disease course showed an improvement in depressive symptoms, regardless of bipolar type. PMID- 19809012 TI - A 66-year-old man with an abdominal aortic aneurysm: review of screening and treatment. AB - Ruptured abdominal aortic aneurysm (AAA) is a common cause of death. Abdominal aortic aneurysms tend to be asymptomatic until the time of rupture, which has a mortality rate of greater than 80%. Therefore, elective repair prior to rupture is preferred if life expectancy is reasonable and the risk of rupture outweighs the risk of repair. Mr F, a 66-year-old man with a 5.2-cm AAA, illustrates the issues surrounding monitoring and treating AAA. Risk factors for AAA include older age, male sex, smoking history, and a family history of AAA. Screening for AAA with ultrasound has been shown to prevent rupture, prevent AAA-related death, and be cost-effective. Risk factors for rupture include larger diameter, female sex, and smoking history. Endovascular repair has lower operative mortality and complications and has replaced standard open surgery in more than half of patients. However, long-term survival is similar after endovascular and open surgical repair. Those at risk of AAA who would benefit from repair should undergo screening. PMID- 19809013 TI - The cover. Granada woman. PMID- 19809014 TI - A piece of my mind. Separation anxiety. PMID- 19809015 TI - Growing calls in United States, Europe to improve regulation of genetic testing. PMID- 19809016 TI - New focus on long-term mental health of patients who survive serious illness. PMID- 19809017 TI - Scientists probe method to prevent inherited mitochondrial gene diseases. PMID- 19809018 TI - Acid-suppressive medication and hospital-acquired pneumonia. PMID- 19809019 TI - Acid-suppressive medication and hospital-acquired pneumonia. PMID- 19809020 TI - Acid-suppressive medication and hospital-acquired pneumonia. PMID- 19809021 TI - Human information processing, health information technology, and medical outcomes. PMID- 19809022 TI - Childhood course of lung function in survivors of bronchopulmonary dysplasia. PMID- 19809023 TI - Bovine lactoferrin supplementation for prevention of late-onset sepsis in very low-birth-weight neonates: a randomized trial. AB - CONTEXT: Sepsis is a common and severe complication in premature neonates, particularly those with very low birth weight (VLBW) (<1500 g). Whether lactoferrin, a mammalian milk glycoprotein involved in innate immune host defenses, can reduce the incidence of sepsis is unknown. In animal models, the probiotic Lactobacillus rhamnosus GG (LGG) enhances the activity of lactoferrin but has not been studied in human infants. OBJECTIVE: To establish whether bovine lactoferrin (BLF), alone or in combination with LGG, reduces the incidence of late-onset sepsis in VLBW neonates. DESIGN, SETTING, AND PATIENTS: Prospective, multicenter, double-blind, placebo-controlled, randomized trial conducted in 11 Italian tertiary neonatal intensive care units. Patients were 472 VLBW infants enrolled from October 1, 2007, through July 31, 2008, and assessed until discharge for development of sepsis. INTERVENTION: Infants were randomly assigned to receive orally administered BLF (100 mg/d) alone (n = 153), BLF plus LGG (6 x 10(9) colony-forming units/d) (n = 151), or placebo (n = 168) from birth until day 30 of life (day 45 for neonates <1000 g at birth). MAIN OUTCOME MEASURE: First episode of late-onset sepsis, ie, sepsis occurring more than 72 hours after birth with isolation of any pathogen from blood or from peritoneal or cerebrospinal fluid. RESULTS: Demographic, clinical, and management characteristics of the 3 groups were similar, including type of feeding and intake of maternal milk. Incidence of late-onset sepsis was significantly lower in the BLF and BLF plus LGG groups (9/153 [5.9%] and 7/151 [4.6%], respectively) than in the control group receiving placebo (29/168 [17.3%]) (risk ratio, 0.34; 95% confidence interval, 0.17-0.70; P = .002 for BLF vs control and risk ratio, 0.27; 95% confidence interval, 0.12-0.60; P < .001 for BLF plus LGG vs control). The decrease occurred for both bacterial and fungal sepsis. No adverse effects or intolerances to treatment occurred. CONCLUSION: Compared with placebo, BLF supplementation alone or in combination with LGG reduced the incidence of a first episode of late-onset sepsis in VLBW neonates. TRIAL REGISTRATION: isrctn.org Identifier: ISRCTN53107700. PMID- 19809024 TI - Association between CYP2D6 polymorphisms and outcomes among women with early stage breast cancer treated with tamoxifen. AB - CONTEXT: The growth inhibitory effect of tamoxifen, which is used for the treatment of hormone receptor-positive breast cancer, is mediated by its metabolites, 4-hydroxytamoxifen and endoxifen. The formation of active metabolites is catalyzed by the polymorphic cytochrome P450 2D6 (CYP2D6) enzyme. OBJECTIVE: To determine whether CYP2D6 variation is associated with clinical outcomes in women receiving adjuvant tamoxifen. DESIGN, SETTING, AND PATIENTS: Retrospective analysis of German and US cohorts of patients treated with adjuvant tamoxifen for early stage breast cancer. The 1325 patients had diagnoses between 1986 and 2005 of stage I through III breast cancer and were mainly postmenopausal (95.4%). Last follow-up was in December 2008; inclusion criteria were hormone receptor positivity, no metastatic disease at diagnosis, adjuvant tamoxifen therapy, and no chemotherapy. DNA from tumor tissue or blood was genotyped for CYP2D6 variants associated with reduced (*10, *41) or absent (*3, *4, *5) enzyme activity. Women were classified as having an extensive (n=609), heterozygous extensive/intermediate (n=637), or poor (n=79) CYP2D6 metabolism. MAIN OUTCOME MEASURES: Time to recurrence, event-free survival, disease-free survival, and overall survival. RESULTS: Median follow-up was 6.3 years. At 9 years of follow up, the recurrence rates were 14.9% for extensive metabolizers, 20.9% for heterozygous extensive/intermediate metabolizers, and 29.0% for poor metabolizers, and all-cause mortality rates were 16.7%, 18.0%, and 22.8%, respectively. Compared with extensive metabolizers, there was a significantly increased risk of recurrence for heterozygous extensive/intermediate metabolizers (time to recurrence adjusted hazard ratio [HR], 1.40; 95% confidence interval [CI], 1.04-1.90) and for poor metabolizers (time to recurrence HR, 1.90; 95% CI, 1.10-3.28). Compared with extensive metabolizers, those with decreased CYP2D6 activity (heterozygous extensive/intermediate and poor metabolism) had worse event-free survival (HR, 1.33; 95% CI, 1.06-1.68) and disease-free survival (HR, 1.29; 95% CI, 1.03-1.61), but there was no significant difference in overall survival (HR, 1.15; 95% CI, 0.88-1.51). CONCLUSION: Among women with breast cancer treated with tamoxifen, there was an association between CYP2D6 variation and clinical outcomes, such that the presence of 2 functional CYP2D6 alleles was associated with better clinical outcomes and the presence of nonfunctional or reduced-function alleles with worse outcomes. PMID- 19809025 TI - Using effectiveness and cost-effectiveness to make drug coverage decisions: a comparison of Britain, Australia, and Canada. AB - CONTEXT: National public insurance for drugs is often based on evidence of comparative effectiveness and cost-effectiveness. This study describes how that evidence has been used across 3 jurisdictions (Australia, Canada, and Britain) that have been at the forefront of evidence-based coverage internationally. OBJECTIVES: To describe how clinical and cost-effectiveness evidence is used in coverage decisions both within and across jurisdictions and to identify common issues in the process of evidence-based coverage. DESIGN, SETTING, AND PARTICIPANTS: Descriptive analysis of retrospective data from the Common Drug Review (CDR) of Canada, National Institute for Health and Clinical Excellence (NICE) in Britain, and Pharmaceutical Benefits Advisory Committee (PBAC) of Australia. All publicly available information as of December 31, 2008, was gathered from each committee's Web site (data set begins in January 2004 [CDR], February 2001 [NICE], and July 2005 [PBAC]). MAIN OUTCOME MEASURE: Listing recommendations for each drug by disease indication. RESULTS: NICE recommended 87.4% (174/199) of submissions for listing compared with a listing rate of 49.6% (60/121) and 54.3% (153/282) for the CDR and PBAC, respectively. Significant uncertainty around clinical effectiveness, typically resulting from inadequate study design or the use of inappropriate comparators and unvalidated surrogate end points, was identified as a key issue in coverage decisions. Recommendations varied considerably across countries, possibly because of differences in the medications reviewed; different agency processes, including the willingness to negotiate on price; and the approach to "me too" drugs. The data suggest that the 3 agencies make recommendations that are consistent with evidence on effectiveness and cost-effectiveness but that other factors are often important. CONCLUSIONS: NICE, PBAC, and CDR face common issues with respect to the quality and strength of the experimental evidence in support of a clinically meaningful effect. However, comparative effectiveness and cost-effectiveness, along with other relevant factors, can be used by national agencies to support drug decision making. The results of the evaluation process in different countries are influenced by the context, agency processes, ability to engage in price negotiation, and perhaps differences in social values. PMID- 19809026 TI - Effects of iron supplementation and depletion on hypoxic pulmonary hypertension: two randomized controlled trials. AB - CONTEXT: Hypoxia is a major cause of pulmonary hypertension in respiratory disease and at high altitude. Recent work has established that the effect of hypoxia on pulmonary arterial pressure may depend on iron status, possibly acting through the transcription factor hypoxia-inducible factor, but the pathophysiological and clinical importance of this interaction is unknown. OBJECTIVE: To determine whether increasing or decreasing iron availability modifies altitude-induced hypoxic pulmonary hypertension. DESIGN, SETTING, AND PARTICIPANTS: Two randomized, double-blind, placebo-controlled protocols conducted in October-November 2008. In the first protocol, 22 healthy sea-level resident men (aged 19-60 years) were studied over 1 week of hypoxia at Cerro de Pasco, Peru (altitude 4340 m). In the second protocol, 11 high-altitude resident men (aged 30-59 years) diagnosed with chronic mountain sickness were studied over 1 month of hypoxia at Cerro de Pasco, Peru. INTERVENTION: In the first protocol, participants received intravenous infusions of Fe(III)-hydroxide sucrose (200 mg) or placebo on the third day of hypoxia. In the second protocol, patients underwent staged isovolemic venesection of 2 L of blood. Two weeks later, patients received intravenous infusions of Fe(III)-hydroxide sucrose (400 mg) or placebo, which were subsequently crossed over. MAIN OUTCOME MEASURE: Effect of varying iron availability on pulmonary artery systolic pressure (PASP) assessed by Doppler echocardiography. RESULTS: In the sea-level resident protocol, approximately 40% of the pulmonary hypertensive response to hypoxia was reversed by infusion of iron, which reduced PASP by 6 mm Hg (95% confidence interval [CI], 4-8 mm Hg), from 37 mm Hg (95% CI, 34-40 mm Hg) to 31 mm Hg (95% CI, 29-33 mm Hg; P = .01). In the chronic mountain sickness protocol, progressive iron deficiency induced by venesection was associated with an approximately 25% increase in PASP of 9 mm Hg (95% CI, 4-14 mm Hg), from 37 mm Hg (95% CI, 30-44 mm Hg) to 46 mm Hg (95% CI, 40-52 mm Hg; P = .003). During the subsequent crossover period, no acute effect of iron replacement on PASP was detected. CONCLUSION: Hypoxic pulmonary hypertension may be attenuated by iron supplementation and exacerbated by iron depletion. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00952302. PMID- 19809027 TI - FDA regulation of tobacco: politics, law, and the public's health. PMID- 19809028 TI - Innovation networks: a strategy to transform primary health care. PMID- 19809029 TI - Applying health care reform principles to mental health and substance abuse services. PMID- 19809030 TI - Disruption and innovation in health care. PMID- 19809031 TI - Lactoferrin supplementation to prevent nosocomial infections in preterm infants. PMID- 19809032 TI - JAMA patient page. Pulmonary hypertension. PMID- 19809037 TI - Medical questionnaires are copyrighted to ensure that validity is maintained. PMID- 19809038 TI - Medical uncertainty: informing decision making for patients with acute pulmonary embolism. PMID- 19809039 TI - Filling the heart failure management void with positive airway pressure. PMID- 19809040 TI - Keeping the workplace safe in troubled times. PMID- 19809041 TI - The patient-ventilator interaction has a third player: the endotracheal tube. PMID- 19809042 TI - Improving access to specialty care for chronic cough. PMID- 19809044 TI - Point: The Texas advance directives act effectively and ethically resolves disputes about medical futility. PMID- 19809045 TI - Nutritional support is maintained: case report. PMID- 19809046 TI - Counterpoint: The Texas advance directives act is ethically flawed: medical futility disputes must be resolved by a fair process. PMID- 19809049 TI - A-lines and B-lines: lung ultrasound as a bedside tool for predicting pulmonary artery occlusion pressure in the critically ill. AB - BACKGROUND: The risk of pulmonary edema is the main limiting factor in fluid therapy in the critically ill. Interstitial edema is a subclinical step that precedes alveolar edema. This study assesses a bedside tool for detecting interstitial edema, lung ultrasound. The A-line is a horizontal artifact indicating a normal lung surface. The B-line is a kind of comet-tail artifact indicating subpleural interstitial edema. The relationship between anterior interstitial edema detected by lung ultrasound and the pulmonary artery occlusion pressure (PAOP) value was investigated. METHOD: We performed a prospective study in medicosurgical ICUs of university-affiliated teaching hospitals. We enrolled 102 consecutive mechanically ventilated patients who all underwent pulmonary artery catheterization. We defined A-predominance as a majority of anterior A lines and B-predominance as a majority of anterior B-lines. These patterns were correlated with PAOP. RESULTS: For diagnosing PAOP 5 years. We present six well characterized cases of arthritis-associated pseudochylothorax, each notable due to their minimal pleural thickening. The median duration of symptoms (or arthritis, in the case of asymptomatic effusions) was 15 months. Such findings cast significant doubt on the conventional concepts of the pathogenesis of rheumatoid-associated pseudochylothorax. Clinicians should consider pseudochylothorax even in short-duration nonfibrotic pleural effusions. PMID- 19809058 TI - Interventional management of pleural infections. AB - Pleural infections represent an important group of disorders that is characterized by the invasion of pathogens into the pleural space and the potential for rapid progression to frank empyema. Previous epidemiologic studies have indicated that empyema is increasing in prevalence, which underscores the importance of urgent diagnosis and effective drainage to improve clinical outcomes. Unfortunately, limited evidence exists to guide clinicians in selecting the ideal drainage intervention for a specific patient because of the broad variation that exists in the intrapleural extent of infection, presence of locules, comorbid features, respiratory status, and virulence of the underlying pathogen. Moreover, many patients experience delays in both the recognition of infected pleural fluid and the initiation of appropriate measures to drain the pleural space. The present review provides an update on the pathogenesis and interventional therapy of pleural infections with an emphasis on the unique role of image-guided drainage with small-bore catheters. PMID- 19809059 TI - A 67-year-old woman with a remote history of breast cancer who presents with dyspnea and chest pain. PMID- 19809060 TI - A 27-year-old man with pleuritic chest pain and hemoptysis after a rugby game. PMID- 19809061 TI - A 45-year-old man with a history of hepatitis C and testicular cancer presents with cavitary lung lesions and palpable purpura. PMID- 19809062 TI - Dangers in using translated medical questionnaires: the importance of conceptual equivalence across languages and cultures in patient-reported outcome measures. PMID- 19809068 TI - Pulmonary venoocclusive disease and failure of specific therapy. PMID- 19809069 TI - Longitudinal assessment of spirometry in World Trade Center responders. PMID- 19809070 TI - Predictors of success in pulmonary rehabilitation for patients with interstitial lung disease. PMID- 19809072 TI - Flawed comparative groups lead to flawed conclusions. PMID- 19809075 TI - Bronchoscopy in China: the Chinese Society of Respiratory Diseases Survey. PMID- 19809076 TI - Could fiberoptic bronchoscopy and CT lung scan differentiate ventilator associated tracheobronchitis from ventilator-associated pneumonia? PMID- 19809077 TI - Trees don't grow in the lungs! PMID- 19809078 TI - Carriage classification of pneumonia rather than time improves survival. PMID- 19809079 TI - Chronic necrotizing pulmonary aspergillosis or invasive pulmonary aspergillosis. PMID- 19809080 TI - Exercise-induced bronchospasm: coding and billing for physician services. PMID- 19809081 TI - Does biofilm formation play a role in ventilator-associated tracheobronchitis? PMID- 19809083 TI - Generically speaking. AB - It's best to use the generic name when referring to a medicine. This reduces the likelihood of accidental double prescribing or purchase of a drug, particularly if it has more than one brand name. Prescribing by generic name is also generally cheaper and avoids confusion because the name is internationally recognised. Nevertheless, brand names enjoy undue prominence, even allowing for the clinical situations where it is important to prescribe by brand rather than generic name. PMID- 19809082 TI - Unify the evaluative procedures and involve peers for increasing use of guidelines in daily practices. PMID- 19809084 TI - Does bnp testing aid diagnosis of heart failure? AB - Heart failure is common, causes major disability and often shortens life. Two UK guidelines advocate the measurement of plasma concentrations of B-type natriuretic peptide (BNP) in diagnosis of chronic heart failure, either in combination with, or as an alternative to, an ECG.1,2 Here we review the evidence for BNP testing in the diagnosis of chronic heart failure, and discuss the implications in terms of availability of the test. PMID- 19809085 TI - Imiquimod for superficial and in situ skin malignancy. AB - Imiquimod, an immune response modifier, is marketed as Aldara 5% cream (Meda) and licensed for treating adults with small superficial basal cell carcinomas (BCCs).1 Numerous uses outside the licensed indications (i.e. 'off-label') have been proposed and practised, including as treatment for pre-cancerous conditions such as Bowen's disease (squamous cell carcinoma in situ) and lentigo maligna (an in situ precursor of melanoma).2,3 Here we review the use of imiquimod for small superficial primary BCC in adults, Bowen's disease and lentigo maligna. PMID- 19809086 TI - Anticoagulation with {blacktriangledown}dabigatran or{blacktriangledown}rivaroxaban. AB - Anticoagulants such as low molecular weight heparins (LMWH; e.g. enoxaparin) and black triangle downfondaparinux are cost-effective measures for reducing the likelihood of venous thromboembolism (VTE) in patients undergoing surgery.1 black triangle downDabigatran etexilate (Pradaxa-Boehringer Ingelheim)2 and black triangle downrivaroxaban (Xarelto - Bayer) are fixed-dose oral anticoagulants licensed in the European Union (EU) for the prevention of VTE in adults who have undergone or are undergoing elective replacement of the hip or knee joints.2,3 Here, we consider their place for these indications. PMID- 19809088 TI - Apoptosis: calling time on apoptosome activity. AB - Apoptosis is a controlled form of cellular demolition, catalyzed by a family of cysteine proteases called caspases. In response to diverse proapoptotic stimuli, caspase-9 is recruited and activated within an oligomeric complex called the apoptosome. The apoptosome drives autocatalytic processing of caspase-9, triggering a proteolytic caspase cascade that results in the biochemical and morphological changes characteristic of cell death. It is unclear why caspase-9 undergoes autocatalytic processing following apoptosome recruitment, because interdomain processing is dispensable for caspase-9 activity. A study has shed light on this issue by demonstrating that caspase-9 processing within the apoptosome promotes its displacement from the complex, leading to inactivation of this protease. Thus, autoprocessing of caspase-9 within the apoptosome serves as a "molecular timer" that limits the proteolytic activity of this complex through displacement of bound caspase-9 molecules. This timer mechanism may enable cells to prevent low amounts of apoptosome activation from spiraling out of control unless sufficient numbers of apoptosomes are assembled within a particular time window, which would drive full-blown caspase activation and apoptosis. PMID- 19809089 TI - Snapshots form a big picture of guanine nucleotide exchange. AB - Small guanosine triphosphatases (GTPases) regulate a vast array of cellular functions. Their highly controlled activation, which is catalyzed by guanine nucleotide exchange factors (GEFs), links input signals emanating from various events such as stimulation of cell surface receptors to a similarly diverse range of downstream responses. Due to the central role of GEFs in the regulation of GTPase-mediated signaling processes, their mode of action has been intensively investigated. A new structural study on the DOCK family of Rho family-specific GEFs now uncovers an unusual variation in the way in which GEFs can regulate the nucleotide status of GTPases and provides the most complete picture of a GDP-GTP exchange cycle to date. PMID- 19809090 TI - Wingless promotes proliferative growth in a gradient-independent manner. AB - Morphogens form concentration gradients that organize patterns of cells and control growth. It has been suggested that, rather than the intensity of morphogen signaling, it is its gradation that is the relevant modulator of cell proliferation. According to this view, the ability of morphogens to regulate growth during development depends on their graded distributions. Here, we describe an experimental test of this model for Wingless, one of the key organizers of wing development in Drosophila. Maximal Wingless signaling suppresses cellular proliferation. In contrast, we found that moderate and uniform amounts of exogenous Wingless, even in the absence of endogenous Wingless, stimulated proliferative growth. Beyond a few cell diameters from the source, Wingless was relatively constant in abundance and thus provided a homogeneous growth-promoting signal. Although morphogen signaling may act in combination with as yet uncharacterized graded growth-promoting pathways, we suggest that the graded nature of morphogen signaling is not required for proliferation, at least in the developing Drosophila wing, during the main period of growth. PMID- 19809091 TI - Coordinated responses to oxygen and sugar deficiency allow rice seedlings to tolerate flooding. AB - Flooding is a widespread natural disaster that leads to oxygen (O(2)) and energy deficiency in terrestrial plants, thereby reducing their productivity. Rice is unusually tolerant to flooding, but the underlying mechanism for this tolerance has remained elusive. Here, we show that protein kinase CIPK15 [calcineurin B like (CBL)-interacting protein kinase] plays a key role in O(2)-deficiency tolerance in rice. CIPK15 regulates the plant global energy and stress sensor SnRK1A (Snf1-related protein kinase 1) and links O(2)-deficiency signals to the SnRK1-dependent sugar-sensing cascade to regulate sugar and energy production and to enable rice growth under floodwater. Our studies contribute to understanding how rice grows under the conditions of O(2) deficiency necessary for growing rice in irrigated lowlands. PMID- 19809092 TI - Sensorimotor cortical activation in patients with cervical spinal cord injury with persisting paralysis. AB - BACKGROUND: It is well documented that cortical sensorimotor representations are altered following nervous system pathology. However, little is known about these representations over time and, more specifically, in paralyzed individuals. OBJECTIVE: To investigate the temporal changes in sensorimotor cortical activation in paralyzed individuals following spinal cord injury (SCI). METHODS: Functional MRI (fMRI) was used to study 4 tetraplegic individuals repeatedly over the first year following traumatic SCI as well as 7 healthy individuals, 3 repeatedly. During fMRI, controls performed ankle movements, and patients attempted them. Standard clinical measures of SCI were used to assess movement ability. RESULTS: Shortly after SCI, activation within the primary motor cortex (M1) was present at levels similar to those in controls. Extensive associated cortical sensorimotor activation, not seen in controls, was present. Over time, as paralysis persisted, activation in M1 was significantly reduced and progressively decreased in associated cortical sensorimotor areas. No session specific dependence in M1 or associated sensorimotor cortical activation was found in healthy individuals. CONCLUSIONS: These findings provide the first report of the temporal evolution of cortical sensorimotor fMRI activation following traumatic SCI in humans who do not recover movement. Coupled with findings in patients who recover post-SCI, our results suggest an association between motor task-related fMRI activation and degree of motor function postinjury. Understanding the time course of plasticity and the relationship between cortical sensorimotor activation and motor ability following SCI could allow assessment of rehabilitation potential, monitoring of therapeutic efficacy, and improvement in therapeutic intervention along the course of recovery. PMID- 19809093 TI - Finding chimeras: a bioinformatics strategy for identification of cross-linked peptides. AB - Chemical cross-linking, followed by identification of the cross-linked residues, is a powerful approach to probe the topologies and interacting surfaces of protein assemblies. In this work, we demonstrate a new bioinformatics approach using multiple program modules within the software package "Protein Prospector" that greatly facilitates the discovery of cross-linked peptides in chemical cross linking studies. Examples are given for how this approach has been used for defining interfaces in heterodimeric and homodimeric protein complexes, both of which provide results in close agreement with crystal structures, verifying the reliability of the approach. PMID- 19809094 TI - HIV-negative status is associated with very early onset of lactation among Ghanaian women. AB - This is a longitudinal cohort study investigating the association between maternal HIV status and the reported onset of lactation. The Research to Improve Infant Nutrition and Growth project recruited 442 mothers from 3 antenatal clinics in the eastern region of Ghana, based on positive, negative, and unknown HIV status. Onset of lactation was assessed by maternal perception and validated with 2 subsamples: measurement of infant breast milk intake (n = 40) and daily infant weight measurement for 2 weeks (n = 150). Multivariate logistic regression was used to identify predictors of very early onset of lactation (onset of lactation < 6 hours). Predictors of very early onset of lactation include HIV negative status (odds ratio = 2.68; P = .014), multiparity (odds ratio = 2.93; P = .009), vaginal delivery (odds ratio = 2.55; P = .035), and having a male child (odds ratio = 1.86; P = .032). The findings indicate an association between maternal HIV status and very early onset of lactation. PMID- 19809095 TI - Pink or blue ... black and blue? Examining pregnancy as a predictor of intimate partner violence and femicide. AB - This study explores the effect of pregnancy on women's risk of IPV victimization. Previous studies have produced conflicting findings and have been limited by comparing only two categories of IPV. The current study included analyses of the effect of pregnancy on three statuses of IPV: no abuse, nonlethal abuse, and lethal abuse. The data used for this project are taken from the first phase of the Chicago Women's Health Risk Study (CWHRS). Findings suggest that both nonlethal and lethal abuse statuses differ significantly from no abuse in that women's risk of IPV victimization may decrease during pregnancy. PMID- 19809096 TI - Spiritual abuse: an additional dimension of abuse experienced by abused Haredi (ultraorthodox) Jewish wives. AB - This article aims to conceptualize spiritual abuse as an additional dimension to physical, psychological, sexual, and economic abuse. Growing out of an interpretivist participatory action research study in a therapeutic Haredi (Jewish ultraorthodox) group of eight abused women, spiritual abuse has been defined as any attempt to impair the woman's spiritual life, spiritual self, or spiritual well-being, with three levels of intensity: (a) belittling her spiritual worth, beliefs, or deeds; (b) preventing her from performing spiritual acts; and (c) causing her to transgress spiritual obligations or prohibitions. The concept and its typology are illustrated by means of examples from the women's abusive experiences and may be of theoretical and therapeutic worldwide relevance. PMID- 19809097 TI - Sexual assault and current mental health: the role of help-seeking and police response. AB - We examine the extent to which seeking help from social service agencies, family and friends, reporting to the police, or responses by the police might buffer or exacerbate the impact of sexual assault on mental health outcomes among sexual assault victims.The trend in many cases was for help-seeking and police response to exacerbate the impact of sexual assault victimization. With respect to depression, we found that the association of rape penetration was greater among those seeking help from social services and those reporting their victimization to the police. Although arresting the offender appears to be associated with higher levels of depression, it actually results in a lower probability of heavy episodic drinking. PMID- 19809098 TI - Secondary traumatic stress among domestic violence advocates: workplace risk and protective factors. AB - This study identified workplace factors associated with secondary traumatic stress (STS) in a sample of 148 domestic violence advocates working in diverse settings. Findings indicate that coworker support and quality clinical supervision are critical to emotional well-being and that an environment in which there is shared power-that is, respect for diversity, mutuality, and consensual decision making-provides better protection for advocates than more traditional, hierarchical organizational models. Furthermore, shared power emerged as the only workplace variable to significantly predict STS above and beyond individual factors. The discussion includes implications for practice and policy as well as directions for future research. PMID- 19809099 TI - Intimate partner violence victimization and parenting stress: assessing the mediating role of depressive symptoms. AB - Guided by the spillover hypothesis and process model of parenting, this study examined relationships between intimate partner violence (IPV) victimization and parenting characteristics among a sample of 1,153 lower-income women. Hierarchical regression was used to investigate a mediational model examining women's self-reports of physical and psychological IPV, depressive symptoms, and parenting stress. Results suggest that depressive symptoms partially mediate the link between a mother's psychological IPV victimization and later self-reported parenting stress. Findings from this study highlight the importance of including maternal psychopathology and other stress and support variables when assessing parenting outcomes among women with histories of IPV. PMID- 19809100 TI - Promoting the safe and strategic use of technology for victims of intimate partner violence: evaluation of the technology safety project. AB - The Technology Safety Project of the Washington State Coalition Against Domestic Violence was designed to increase awareness and knowledge of technology safety issues for domestic violence victims, survivors, and advocacy staff. The project used a "train-the-trainer" model and provided computer and Internet resources to domestic violence service providers to (a) increase safe computer and Internet access for domestic violence survivors in Washington, (b) reduce the risk posed by abusers by educating survivors about technology safety and privacy, and (c) increase the ability of survivors to help themselves and their children through information technology. Evaluation of the project suggests that the program is needed, useful, and effective. Consumer satisfaction was high, and there was perceived improvement in computer confidence and knowledge of computer safety. Areas for future program development and further research are discussed. PMID- 19809101 TI - A computational study of lambda-lac mutants. AB - We present a comprehensive, computational study of the properties of bacteriophage lambda mutants designed by Atsumi and Little (2006 Proc. Natl. Acad. Sci. 103 4558-63). These phages underwent a genetic reconstruction where Cro was replaced by a dimeric form of the Lac repressor. To clarify the theoretical characteristics of these mutants, we built a detailed thermodynamic model. The mutants all have a different genetic wiring than the wild-type lambda. One group lacks regulation of P(RM) by the lytic protein. These mutants only exhibit the lysogenic equilibrium, with no transiently active P(R). The other group lacks the negative feedback from CI. In this group, we identify a handful of bi-stable mutants, although the majority only exhibit the lysogenic equilibrium. The experimental identification of functional phages differs from our predictions. From a theoretical perspective, there is no reason why only 4 out of 900 mutants should be functional. The differences between theory and experiment can be explained in two ways. Either, the view of the lambda phage as a bi-stable system needs to be revised, or the mutants have in fact not undergone a modular replacement, as intended by Atsumi and Little, but constitute instead a wider systemic change. PMID- 19809102 TI - Statistical LOR estimation for a high-resolution dMiCE PET detector. AB - We develop a statistical line of response (LOR) estimator of the three dimensional interaction positions of a pair of annihilation photons in a PET detector module with depth of interaction capability. The three-dimensional points of interaction of a coincidence pair of photons within the detector module are estimated by calculation of an expectation of the points of interaction conditioned on the signals measured by the photosensors. This conditional expectation is computed from estimates of the probability density function of the light collection process and a model of the kinetics of photon interactions in the detector module. Our algorithm is capable of handling coincidences where each annihilation photon interacts any number of times within the detector module before being completely absorbed or escaping. In the case of multiple interactions, our algorithm estimates the position of the first interaction for each of the coincidence photons. This LOR estimation algorithm is developed for a high-resolution PET detector capable of providing depth-of-interaction information. Depth of interaction is measured by tailoring the light shared between two adjacent detector elements. These light-sharing crystal pairs are referred to as dMiCE and are being developed in our lab. Each detector element in the prototype system has a 2 x 2 mm(2) cross section and is directly coupled to a micro-pixel avalanche photodiode (MAPD). In this set-up, the distribution of the ratio of light shared between two adjacent detector elements can be expressed as a function of the depth of interaction. Monte Carlo experiments are performed using our LOR estimation algorithm and compared with Anger logic. We show that our LOR estimation algorithm provides a significant improvement over Anger logic under a variety of parameters. PMID- 19809103 TI - The dynamics of a non-equilibrium bubble near bio-materials. AB - In many medical treatments oscillating (non-equilibrium) bubbles appear. They can be the result of high-intensity-focused ultrasound, laser treatments or shock wave lithotripsy for example. The physics of such oscillating bubbles is often not very well understood. This is especially so if the bubbles are oscillating near (soft) bio-materials. It is well known that bubbles oscillating near (hard) materials have a tendency to form a high speed jet directed towards the material during the collapse phase of the bubble. It is equally well studied that bubbles near a free interface (air) tend to collapse with a jet directed away from this interface. If the interface is neither 'free' nor 'hard', such as often occurs in bio-materials, the resulting flow physics can be very complex. Yet, in many bio applications, it is crucial to know in which direction the jet will go (if there is a jet at all). Some applications require a jet towards the tissue, for example to destroy it. For other applications, damage due to impacting jets is to be prevented at all cost. This paper tries to address some of the physics involved in these treatments by using a numerical method, the boundary element method (BEM), to study the dynamics of such bubbles near several bio-materials. In the present work, the behaviour of a bubble placed in a water-like medium near various bio-materials (modelled as elastic fluids) is investigated. It is found that its behaviour depends on the material properties (Young's modulus, Poisson ratio and density) of the bio-material. For soft bio-materials (fat, skin, brain and muscle), the bubble tends to split into smaller bubbles. In certain cases, the resulting bubbles develop opposing jets. For hard bio-materials (cornea, cartilage and bone), the bubble collapses towards the interface with high speed jets (between 100 and about 250 m s(-1)). A summary graph is provided identifying the combined effects of the dimensionless elasticity (kappa) and density ratio (alpha) of the elastic materials which will result in a nearby oscillating bubble jetting towards, splitting or jetting away from the elastic material interface. Since the phenomenon of a bubble jetting away from an elastic material as it collapses has not been reported before in the literature, experiments were performed to validate the numerical observation. A bubble is created in a heavy fluid (hydrofluoroether (HFE)) using a laser pulse. The bubble collapses near the elastic material polydimethylsiloxane (PDMS). The experimental results obtained are compared with the corresponding simulation. The simulation provides spatial and temporal details about the bubble dynamics beyond experimental limits and can therefore be considered as a very useful tool to get a better understanding of the physics involved. PMID- 19809104 TI - Asymmetric hysteresis loop in magnetostatic-biased multilayer nanowires. AB - The hysteresis of multilayer nanowires composed by a soft magnetic cylindrical wire, a non-magnetic spacer layer and an external hard magnetic shell is investigated. The external magnetic shell originates a non-homogeneous magnetic field on the inner wire, which is responsible for a displacement and a change of the width of the hysteresis curve of the wire. Moreover, different reversal modes occur at each branch of the hysteresis loop, which can be understood by analyzing the interaction magnetostatic field along the wire. Our results open the possibility of controlling two parameters of the hysteresis loop, the coercivity and the bias, providing an interesting system to be investigated. PMID- 19809105 TI - Mass fabrication of resistive random access crossbar arrays by step and flash imprint lithography. AB - Step and flash imprint lithography (SFIL) is a promising method recently used for next generation lithographic technology because it is a high-speed process that can be carried out at room temperature and low pressures. Improvements made to SFIL enable the replication of crossbar patterns with a high resolution and the development of suitable materials and techniques to achieve high resolution capability. In this study, SFIL is used to fabricate high-density random access crossbar arrays based on a NiO resistive switching system. The bottom and top electrodes are transferred onto silicon wafers perpendicular to each electrode using the inductively coupled plasma reactive ion etching (ICP-RIE) technique. Direct metal etching without a wet-based process minimizes damage to the electrode surface. The I-V curves of individual active cells (70 x 70 nm(2)) for crossbar arrays reveal the unipolar resistive switching (RS) behaviour of the fabricated device. A high off/on resistance ratio (>10(4)) and reproducible resistance switching characteristics for each active cell were found in different fields and for different wafers. The experimental data indicate that high-density crossbar arrays can be well replicated and that the electrical performance of these arrays is reliable. PMID- 19809106 TI - Reduction and oxidation of oxide ion conductors with conductive atomic force microscopy. AB - Local accumulation and dissipation of charges on the surface of oxide ion conductors resulting from electric potentials were observed with conductive atomic force microscopy (AFM). After a negative bias was applied at the tip, a sequence of surface potential maps appeared compatible with electron injection onto the electrolyte surface. Applying a positive bias, in contrast, generated a positive surface charge adjacent to the tip contact area. This observation is consistent with the formation of oxide ion vacancies on the oxide surface. In addition, oxide ion conductivity at a low temperature range (100-200 degrees C) was obtained, and the activation energy for diffusion in gadolinia-doped ceria (GDC) was calculated as approximately 0.56 eV, implying that the majority of oxide ion vacancies diffuse on the surface rather than inside the bulk of the electrolyte. PMID- 19809107 TI - Reduced graphene oxide for room-temperature gas sensors. AB - We demonstrated high-performance gas sensors based on graphene oxide (GO) sheets partially reduced via low-temperature thermal treatments. Hydrophilic graphene oxide sheets uniformly suspended in water were first dispersed onto gold interdigitated electrodes. The partial reduction of the GO sheets was then achieved through low-temperature, multi-step annealing (100, 200, and 300 degrees C) or one-step heating (200 degrees C) of the device in argon flow at atmospheric pressure. The electrical conductance of GO was measured after each heating cycle to interpret the level of reduction. The thermally-reduced GO showed p-type semiconducting behavior in ambient conditions and was responsive to low concentration NO2 and NH3 gases diluted in air at room temperature. The sensitivity can be attributed mainly to the electron transfer between the reduced GO and adsorbed gaseous molecules (NO2/NH3). Additionally, the contact between GO and the Au electrode is likely to contribute to the overall sensing response because of the adsorbates-induced Schottky barrier variation. A simplified model is used to explain the experimental observations. PMID- 19809108 TI - The fabrication of diversiform nanostructure forests based on residue nanomasks synthesized by oxygen plasma removal of photoresist. AB - A simple lithography-free approach for fabricating diversiform nanostructure forests is presented. The key technique of the approach is that randomly distributed nanoscale residues can be synthesized on substrates simply by removing photoresist with oxygen plasma bombardment. These nanoresidues can function as masks in the subsequent etching process for nanopillars. By further spacer and then deep etching processes, a variety of forests composed of regular, tulip-like or hollow-head nanopillars as well as nanoneedles are successfully achieved in different etching conditions. The pillars have diameters of 30-200 nm and heights of 400 nm-3 microm. The needles reach several microns in height, with their tips less than 10 nm in diameter. Moreover, microstructures containing these nanostructure forests, such as surface microchannels, have also been fabricated. This approach is compatible with conventional micro/nano electromechanical system (MEMS/NEMS) fabrication. PMID- 19809109 TI - Dynamic susceptibility evidence of surface spin freezing in ultrafine NiFe2O4 nanoparticles. AB - We investigated the dynamic behavior of ultrafine NiFe2O4 nanoparticles (average size D = 3.5 nm) that exhibit anomalous low temperature magnetic properties such as low saturation magnetization and high-field irreversibility in both M(H) and ZFC-FC processes. Besides the expected blocking of the superspin, observed at T1 approximately 45 K, the system undergoes a magnetic transition at T2 approximately 6 K. For the latter, frequency- and temperature-resolved dynamic susceptibility data reveal characteristics that are unambiguously related to collective spin freezing: the relative variation (per frequency decade) of the in phase susceptibility peak temperature is approximately 0.025, critical dynamics analysis yields an exponent znu = 9.6 and a zero-field freezing temperature T(F) = 5.8 K, and, in a magnetic field, T(F)(H) is excellently described by the de Almeida-Thouless line delta T(F) = 1 - T(F)(H)/T(F) alpha H(2/3). Moreover, out of-phase susceptibility versus temperature datasets collected at different frequencies collapse on a universal dynamic scaling curve. All these observations indicate the existence of a spin-glass-like surface layer that surrounds the superparamagnetic core and undergoes a transition to a frozen state upon cooling below 5.8 K. PMID- 19809110 TI - A self-sensing carbon nanotube/cement composite for traffic monitoring. AB - In this paper, a self-sensing carbon nanotube (CNT)/cement composite is investigated for traffic monitoring. The cement composite is filled with multi walled carbon nanotubes whose piezoresistive properties enable the detection of mechanical stresses induced by traffic flow. The sensing capability of the self sensing CNT/cement composite is explored in laboratory tests and road tests. Experimental results show that the fabricated self-sensing CNT/cement composite presents sensitive and stable responses to repeated compressive loadings and impulsive loadings, and has remarkable responses to vehicular loadings. These findings indicate that the self-sensing CNT/cement composite has great potential for traffic monitoring use, such as in traffic flow detection, weigh-in-motion measurement and vehicle speed detection. PMID- 19809111 TI - Modulating the optical and electronic properties of highly symmetric Si quantum dots. AB - The optical and electronic properties of quantum dots are critically influenced by the electronic states around the last occupied first empty levels. Using density functional theory calculations we show three alternative ways to alter the optical response of highly stable, highly symmetric hydrogen-terminated fullerene and polycrystalline Si quantum dots: small shape changes, applied uniaxial strain and exohedral doping. The demonstrated effectiveness renders symmetry lowering, the common theme of these processes, as a promising concept for deliberately adjusting optical response. PMID- 19809112 TI - High room-temperature photoluminescence of one-dimensional Ta2O5 nanorod arrays. AB - In this study we analyzed the structural and electronic properties of a new morphological form, one-dimensional (1D) Ta2O5 nanorod arrays, which were synthesized by hot filament metal vapor deposition. Field-emission scanning electron microscopy (FESEM) showed the 1D Ta2O5 nanorods to be arranged in a large-area high-density array about 50 nm wide and approximately 550 nm long. X ray photoemission spectroscopy (XPS) revealed not only the electronic structures and chemical properties of the 1D Ta2O5 nanorods but also their stoichiometric Ta and O compositions. Photoluminescence (PL) spectra showed intensive green-light, yellow-light and red-light emissions at room temperature. These emissions simultaneously emerged from the trap levels of oxygen vacancies within the Ta2O5 bandgap. The emission results strongly indicate that the 1D Ta2O5 nanorods are good room-temperature visible-light emitters. PMID- 19809113 TI - A study of tungsten oxide nanowires self-organized on mica support. AB - Self-organized straight nanowires of WO3 have been epitaxially grown on muscovite mica in low super-saturation conditions. Morphology, structure and chemical composition of the prepared nanostructures have been investigated by scanning electron microscopy (SEM), x-ray diffraction (XRD) and x-ray photoelectron spectroscopy (XPS). SEM permits us to observe nanowire networks and substrate electron channeling patterns (ECP) simultaneously and thus to determine crystallographic direction of nanowire growth. The experimental results show that straight WO3 nanowires are orientated preferentially parallel to two of three high symmetry crystallographic directions of mica [100] and [110] or [100] and [Formula: see text]. XRD and XPS measurements indicated self-assembly of very thin nanowires of hexagonal tungsten bronze in the first stage of growth followed by the formation of stoichiometric WO3. The growth mechanism has been studied as a function of different preparation conditions with special focus on the role of potassium ions present on the mica surface. Based on obtained results a growth model of tungsten oxide nanowires on mica is proposed. PMID- 19809114 TI - Hybrid light-emitting diodes based on flexible sheets of mass-produced ZnO nanowires. AB - We report the production of free-standing thin sheets made up of mass-produced ZnO nanowires and the application of these nanowire sheets for the fabrication of ZnO/organic hybrid light-emitting diodes in the manner of assembly. Different p type organic semiconductors are used to form heterojunctions with the ZnO nanowire film. Electroluminescence measurements of the devices show UV and visible emissions. Identical strong red emission is observed independent of the organic semiconductor materials used in this work. The visible emissions corresponding to the electron transition between defect levels within the energy bandgap of ZnO are discussed. PMID- 19809115 TI - Superstructures of PbS nanocrystals in a conjugated polymer and the aligning role of oxidation. AB - We present a method to directly align PbS nanocrystals in micron-sized superstructures within a conjugated polymer. First, lead sulfide nanocrystals are directly synthesized in a MEH-PPV suspension via a single pot, surfactant-free method. Post-synthesis precipitation of the composite solution involving mild oxidation of the nanocrystals results in the formation of nanocrystal-polymer and nanocrystal-oxide superstructures. Detailed TEM is used to study the crystallographic nature of these structures and the roles of polymer and lead sulfate. An epitaxial relationship between lead sulfide and lead sulfate at the nanoscale is shown, giving insight into the oxidation rates of the PbS nanocrystals' facets. PMID- 19809116 TI - The fabrication of short metallic nanotubes by templated electrodeposition. AB - Template-based electrochemical synthesis has widely been used to produce metal nanowires and nanorods. Commercially available filtration membranes, such as anodic aluminum oxide (AAO) and polycarbonate track etch membranes, have commonly been utilized as hard templates for this purpose. In this process, a thick metal film is usually sputtered or vacuum evaporated onto one side of the membrane to block the pores and serve as the working electrode for the subsequent electrodeposition. Here, we show that during the deposition of the metal electrode for AAO membranes, the electrode metal diffuses into the pores and is deposited on the pore walls which leads to preferential electrodeposition of metal on the walls and therefore forms metal tubes. This phenomenon has been utilized to fabricate short nanotubes by carefully controlling the electrodeposition conditions. The process is a straightforward method for any electroplatable materials to form nanoscale tubular structures. The effects of working electrodes and electrodeposition conditions on the formation of tubular structures are discussed in detail. A new mechanism based on this simple fact is proposed to explain the formation of Ni tubes by Ni-Cu co-deposition. Also, we demonstrate how to distinguish magnetic nanotubes from nanorods by a simple magnetic measurement. PMID- 19809117 TI - The electronic transport properties of ternary Cd(1-x)Zn(x)S nanowire networks. AB - We present the electronic transport characteristics of ternary alloy Cd(1 x)Zn(x)S nanowire networks in the dark and under white light illumination. Compared to the negligible dark current, we observed a photocurrent enhancement of up to four orders of magnitude at an intensity of 460 mW cm(-2). The time constant of the dynamic photoresponse is approximately 5 s. The current-voltage characteristics at different intensities show Ohmic behavior at low bias and space charge limited conduction (SCLC) at higher bias voltages. The SCLC behavior and slow time response indicate that the charge transport is dominated by tunneling at the percolating inter-nanowire junctions. PMID- 19809118 TI - Local conductance measurements of double-layer graphene on SiC substrate. AB - The microscopic structural and electrical properties of few-layer graphene grown on an SiC substrate were characterized by low-energy electron microscopy, transmission electron microscopy and scanning probe microscopy measurements of local conductance. The double-layer graphene sheet was confirmed to be continuous across the atomic steps on the buried SiC substrate surface, and the measured local conductance was clearly modified in the vicinity of the steps. The conductance decreased (slightly increased) at the lower (upper) side of the steps, suggesting deformation-induced strain is the origin of the conductance modification. From the contact force dependence of the conductance images, the effective contact areas for both nanogap-probe and point-probe measurements were estimated. PMID- 19809119 TI - Surface morphology effects on polarization switching in nanoscale ferroelectrics. AB - The effects of surface morphology on polarization switching in thin ferroelectric films are investigated using a real-space, time-dependent Ginzburg-Landau model that incorporates electrostatic interactions. We consider a two-dimensional uni axial ferroelectric film with a thickness that varies sinusoidally. Polarization switching, starting from a single domain remnant state, is simulated for several surface modulation amplitudes and wavelengths. We demonstrate that surface heterogeneities produce inhomogeneities in the electric field within the film. These inhomogeneities become preferential sites for easy nucleation of reverse domains. This has a profound effect on the external field necessary to switch the polarization. Increasing the surface undulation amplitude significantly reduces the coercive field compared to the ideal flat film, even for very small amplitude modulations in the thickness. Although surface roughness decreases the field required to form reverse domains, it also hinders subsequent domain wall migration. In fact, for very high amplitude and small wavelength surface morphologies, complete switching to a single domain state becomes impossible. This is because the domain walls become trapped near the peaks in the modulated surface. The technological implications of the present results for utilization of surface roughness and for surface morphology design are discussed. PMID- 19809120 TI - Topochemical growth of textured polycrystalline barium hexaferrite from oriented antiferromagnetic alpha-FeOOH nanorods. AB - Nanorods of goethite, i.e. alpha-FeOOH, were mixed with BaCO3, dispersed in a polymer solution, and oriented under a 90 kOe magnetic field during polymerization. The orientation arose principally from the interaction of the magnetic field with the anisotropic antiferromagnetism of the goethite particles. The oriented antiferromagnetic particles act as seeds for the topochemical growth of BaFe12O19 ferrite grains along the [0001] direction. The degree of grain orientation was determined using magnetic measurements and orientation distribution functions and pole figures determined by electron backscatter diffraction analysis. PMID- 19809121 TI - Facile and rapid synthesis of highly luminescent nanoparticles via pulsed laser ablation in liquid. AB - This paper demonstrates the usefulness of pulsed laser ablation in liquids as a fast screening synthesis method able to prepare even complex compositions at the nanoscale. Nanoparticles of Y2O3:Eu3+, Lu2O2S:Eu3+, Gd2SiO5:Ce3+, Lu3TaO7:Gd3+ and Tb3+ are successfully synthesized by pulsed laser ablation in liquids. The phase and stoichiometries of the original materials are preserved while the sizes are reduced down to 5-10 nm. The optical properties of the materials are also preserved but show some small variations and some additional structures which are attributed to the specificities of the nanoscale (internal pressure, inhomogeneous broadening, surface states, etc). PMID- 19809122 TI - Trade-off bounds for the Pareto surface approximation in multi-criteria IMRT planning. AB - One approach to multi-criteria IMRT planning is to automatically calculate a data set of Pareto-optimal plans for a given planning problem in a first phase, and then interactively explore the solution space and decide on the clinically best treatment plan in a second phase. The challenge of computing the plan data set is to ensure that all clinically meaningful plans are covered and that as many clinically irrelevant plans as possible are excluded to keep computation times within reasonable limits. In this work, we focus on the approximation of the clinically relevant part of the Pareto surface, the process that constitutes the first phase. It is possible that two plans on the Pareto surface have a small, clinically insignificant difference in one criterion and a significant difference in another criterion. For such cases, only the plan that is clinically clearly superior should be included into the data set. To achieve this during the Pareto surface approximation, we propose to introduce bounds that restrict the relative quality between plans, the so-called trade-off bounds. We show how to integrate these trade-off bounds into the approximation scheme and study their effects. The proposed scheme is applied to two artificial cases and one clinical case of a paraspinal tumor. For all cases, the quality of the Pareto surface approximation is measured with respect to the number of computed plans, and the range of values occurring in the approximation for different criteria is compared. Through enforcing trade-off bounds, the scheme disregards clinically irrelevant plans during the approximation. Thereby, the number of plans necessary to achieve a good approximation quality can be significantly reduced. Thus, trade-off bounds are an effective tool to focus the planning and to reduce computation time. PMID- 19809123 TI - Thermal ablation produced using a surgical toroidal high-intensity focused ultrasound device is independent from hepatic inflow occlusion. AB - In the liver, the efficacy of radiofrequency or high-intensity focused ultrasound (HIFU) ablation is impaired by blood perfusion. This can be overcome by hepatic inflow occlusion. Here we report the in vivo evaluation of ablations performed in the liver using a surgical toroidal HIFU device used during an open procedure with and without hepatic inflow occlusion. The HIFU device was composed of 256 toroidal-shaped emitters working at 3 MHz and an integrated ultrasound imaging probe working at 7.5 MHz. Using an intermittent Pringle maneuver (IPM), thermal ablations were created in three pigs with hepatic inflow occlusion (IPM group) and in three pigs with normal perfusion (NoIPM group). The ablations were studied on sonograms, macroscopically and microscopically 14 days after the treatment. In the NoIPM group, the average coagulated volume obtained after a 40 s exposure was 7.4 +/- 3.8 cm(3) (2.2-16.6). In the IPM group, the average ablated volume was 6.3 +/- 2.9 cm(3) (2.6-12.1). There was no significant difference between the two groups in terms of ablated volume (p = 0.25), diameter (p = 0.37) or depth (p = 0.61). Therefore, a toroidal-shaped HIFU device allows treatment in the liver that can be considered as independent from hepatic inflow occlusion. PMID- 19809124 TI - An extended convection diffusion model for red blood cell-enhanced transport of thrombocytes and leukocytes. AB - Transport phenomena of platelets and white blood cells (WBCs) are fundamental to the processes of vascular disease and thrombosis. Unfortunately, the dilute volume occupied by these cells is not amenable to fluid-continuum modeling, and yet the cell count is large enough that modeling each individual cell is impractical for most applications. The most feasible option is to treat them as dilute species governed by convection and diffusion; however, this is further complicated by the role of the red blood cell (RBC) phase on the transport of these cells. We therefore propose an extended convection-diffusion (ECD) model based on the diffusive balance of a fictitious field potential, Psi, that accounts for the gradients of both the dilute phase and the local hematocrit. The ECD model was applied to the flow of blood in a tube and between parallel plates in which a profile for the RBC concentration field was imposed and the resulting platelet concentration field predicted. Compared to prevailing enhanced-diffusion models that dispersed the platelet concentration field, the ECD model was able to simulate a near-wall platelet excess, as observed experimentally. The extension of the ECD model depends only on the ability to prescribe the hematocrit distribution, and therefore may be applied to a wide variety of geometries to investigate platelet-mediated vascular disease and device-related thrombosis. PMID- 19809126 TI - An optically stimulated luminescence system to measure dose profiles in x-ray computed tomography. AB - This paper describes an LED-based optically stimulated luminescence (OSL) system for dose profile measurements using OSL detector strips and investigates its performance in x-ray computed tomography (CT) dosimetry. To compensate for the energy response of the Al(2)O(3):C OSL detectors, which have an effective atomic number of 11.28, field-specific energy correction factors were determined using two methods: (a) comparing the OSL profiles with ionization chamber point measurements (0.3 cm(3) ionization chamber) and (b) comparing the OSL profiles integrated over a 100 mm length with 100 mm long pencil ionization chamber measurements. These correction factors were obtained for the CT body and head phantoms, central and peripheral positions and three x-ray tube potential differences (100 kVp, 120 kVp and 140 kVp). The OSL dose profiles corrected by the energy dependence agreed with the ionization chamber point measurements over the entire length of the phantom (300 mm). For 120 kVp x-ray tube potential difference, the CTDI(100) values calculated using the OSL dose profiles corrected for the energy dependence and those obtained from an independent measurement with a 100 mm long pencil ionization chamber also agreed within +/-5%. PMID- 19809125 TI - Topographic localization of brain activation in diffuse optical imaging using spherical wavelets. AB - Diffuse optical imaging is a non-invasive technique that uses near-infrared light to measure changes in brain activity through an array of sensors placed on the surface of the head. Compared to functional MRI, optical imaging has the advantage of being portable while offering the ability to record functional changes in both oxy- and deoxy-hemoglobin within the brain at a high temporal resolution. However, the reconstruction of accurate spatial images of brain activity from optical measurements represents an ill-posed and underdetermined problem that requires regularization. These reconstructions benefit from incorporating prior information about the underlying spatial structure and function of the brain. In this work, we describe a novel image reconstruction approach which uses surface-based wavelets derived from structural MRI to incorporate high-resolution anatomical and structural prior information about the brain. This surface-based approach is used to approximate brain activation patterns through the reconstruction and presentation of topographical (two dimensional) maps of brain activation directly onto the folded surface of the cortex. The set of wavelet coefficients is directly estimated by a truncated singular-value decomposition based pseudo-inversion of the wavelet projection of the optical forward model. We use a reconstruction metric based on Shannon entropy which quantifies the sparse loading of the wavelet coefficients and is used to determine the optimal truncation and regularization of this inverse model. In this work, examples of the performance of this model are illustrated for several cases of numerical simulation and experimental data with comparison to functional magnetic resonance imaging. PMID- 19809127 TI - Enhanced ammonium removal from liquid anaerobic digestion residuals in an advanced sequencing batch reactor system. AB - When treating effluents from anaerobic processing of organic wastes, the challenge lies in the large quantity of recalcitrant COD and in the high nitrogen content. The data presented in this study illustrate an advanced SBR concept that is able to significantly reduce the organic load. It was found that undigested mashed bio-waste bypassing the anaerobic digestion is highly suitable as an external carbon source to compensate deficiencies of readily biodegradable BOD required for denitrification. In order to utilize the carbon source as efficiently as possible, the SBR was operated in a mode by which nitrification/denitrification is achieved by a shortcut process using nitrite. This procedure reduced the requirement of an external carbon source to only 5% (v/v). Moreover, through an optimized SBR sequence for an influent concentration of 14,000 mg COD/l and 3,800 mg TN/l, nearly complete removal of nitrogen (>95%) was achieved. The average removal rates for COD and BOD were 83% and 86%, respectively. The applied hydraulic retention time (HRT) of the SBR was 6.8 days. The elevated temperature of 30-41 degrees C in the reactor as a result of the high biological activity was advantageous in terms of enhanced reaction kinetics and also provided better process stability. PMID- 19809128 TI - Internal recycle to improve denitrification in a step feed anoxic/aerobic activated sludge system. AB - During periods of low load (weekends and holidays) the Mangere wastewater treatment plant effluent has breached the summer consent conditions for total nitrogen. The purpose of this research was to determine if an internal recycle would improve nitrogen removal in the anoxic/aerobic activated sludge reactors sufficient to meet the summer resource consent standard. The recycle returned nitrate rich mixed liquor from the downstream aerobic zone back to the initial anoxic zone, thus potentially improving denitrification. A full scale trial showed that installation of the internal recycle on each RC would have satisfied the resource consent for total nitrogen in most cases over the three summer resource consent periods since the upgrade. However, further modifications of the internal recycle would be required to ensure that consent conditions were satisfied at all times and to improve the consistency of the results. PMID- 19809129 TI - Metabolic models to investigate energy limited anaerobic ecosystems. AB - Anaerobic wastewater treatment is shifting from a philosophy of solely pollutants removal to a philosophy of combined resource recovery and waste treatment. Simultaneous wastewater treatment with energy recovery in the form of energy rich products, brings renewed interest to non-methanogenic anaerobic bioprocesses such as the anaerobic production of hydrogen, ethanol, solvents, VFAs, bioplastics and even electricity from microbial fuel cells. The existing kinetic-based modelling approaches, widely used in aerobic and methanogenic wastewater treatment processes, do not seem adequate in investigating such energy limited microbial ecosystems. The great diversity of similar microbial species, which share many of the fermentative reaction pathways, makes quantify microbial groups very difficult and causes identifiability problems. A modelling approach based on the consideration of metabolic reaction networks instead of on separated microbial groups is suggested as an alternative to describe anaerobic microbial ecosystems and in particular for the prediction of product formation as a function of environmental conditions imposed. The limited number of existing relevant fermentative pathways in conjunction with the fact that anaerobic reactions proceed very close to thermodynamic equilibrium reduces the complexity of such approach and the degrees of freedom in terms of product formation fluxes. In addition, energy limitation in these anaerobic microbial ecosystems makes plausible that selective forces associated with energy further define the system activity by favouring those conversions/microorganisms which provide the most energy for growth under the conditions imposed. PMID- 19809130 TI - Removal of organic matter and nitrogen in an horizontal subsurface flow (HSSF) constructed wetland under transient loads. AB - A monitoring campaign in a horizontal subsurface flow constructed wetland under the influence of transient loads of flow-rate, organic matter, nitrogen and suspended solids showed an irregular removal of COD and TSS and lower both removal efficiencies and mass removal rates than the ones observed in other studies for similar operating conditions. This circumstance is associated to the presence of large amount of particulate organic matter from non-point sources. The mass removal rate of ammonia increased 39% as both the water and soil temperatures increased from weeks 1-8 to weeks 9-14. A good correlation between mass load and mass removal rate was observed for all measured parameters, which attests a satisfactory response of the bed under to transient loads. PMID- 19809131 TI - Real-time in-situ measurement of haemoglobin in wastewater. AB - The meat processing industry generates large volumes of relatively high load wastewater. In New Zealand and Australia this wastewater is often pre-treated on site and then discharged to environmental waters or municipal sewers. Owing to the limited number of water quality parameters which can be measured in real-time it is often difficult for industry to optimise treatment processes or public bodies to monitor for water-quality compliance. Abattoir wastewater is often observed to be red in colour, owing to the presence of haemoglobin. Measurement of visible light absorption spectra of wastewater grab samples has for some time provided information about blood concentration. However such grab sampling techniques are piecemeal and cannot provide instantaneous time resolved signals which are required for process control or comprehensive monitoring. In this work an in-situ UV/VIS spectrometer is used to continuously determine the concentration of haemoglobin in wastewater arriving for treatment at two different Wastewater Treatment Plants (WWTPs). The data is of high temporal resolution- data recorded at the distant WWTPs allows for identification process events, such as the end of shift wash downs. PMID- 19809132 TI - Using Geographical Information Systems (GIS) as an instrument of water resource management: a case study from a GIS-based Water Safety Plan in Germany. AB - Following international developments and the new WHO Drinking Water Guidelines (WHO 2004) a process-orientated concept for risk, monitoring and incident management has been developed and implemented in this study. The concept will be reviewed with special consideration for resource protection (first barrier of the multi-barrier system) and in turn, for the Water Safety Plan (WSP) which adequately considers-beyond the current framework of legal requirements-possible new hygienic-microbiologically relevant risks (especially emerging pathogens) for the drinking water supply. The development of a WSP within the framework of risk, monitoring and incident management includes the application of Geographical Information Systems (GIS). In the present study, GIS was used for visualization and spatial analysis in decisive steps in the WSP. The detailed process of GIS supported implementation included the identification of local participants and their tasks and interactions as an essential part of risk management. A detailed ecological investigation of drinking water conditions in the catchment area was conducted in addition to hazard identification, risk assessment and the monitoring of control measures. The main task of our study was to find out in which steps of the WSP the implementation of GIS could be integrated as a useful, and perhaps even an essential tool. PMID- 19809133 TI - Performance assessment of AS-SBR and UF-MBR for hotel wastewater treatment. AB - A large number of tourist structures in Venice (Italy) have small sized on-site treatment systems for their wastewater. Due to its historical characteristics, the city has no public sewerage system and untreated hotel wastewater represents a serious hazard for its lagoon environment. This study focused on the wastewater facilities installed in two hotels adopting an Activated Sludge Sequencing Batch Reactor (AS-SBR) and an Ultra-Filtration Membrane Biological Reactor (UF-MBR). Their performance was checked in terms of both traditional physico-chemical and ecotoxicological parameters, the importance of which has recently been recognised by EU regulatory dispositions and OSPAR indications. Acute and sub-chronic endpoints were both considered on a whole effluent toxicity basis by means of Vibrio fischeri and Crassostrea gigas, respectively. The two months monitoring survey evidenced that the UF-MBR was more efficient than the AS-SBR in providing high-quality discharges under both chemical and ecotoxicological viewpoints. PMID- 19809134 TI - The effect of adapting cellulose degrading microorganisms to 25 degrees C providing energy sources for biological sulphate removal. AB - Vast volumes of Acid Mine Drainage (AMD) are still being generated in South Africa, due to decant from both active and closed mines. Research to find a cost effective, environmentally friendly treatment system to reduce the salinity and to neutralise the acidity of AMD is ongoing. The study presented here showed that high sulphate removal efficiencies were achieved applying the biological treatment technology, thereby using the degradation products of grass-cellulose as the carbon and energy sources. The process was conducted at 25 degrees C, as opposed to 37 degrees C described previously, using a one-stage hybrid reactor system, treating both sulphate rich synthetic feed water and pre-treated AMD. The results showed that the fermentation microbes, originating from rumen fluid, derived from cattle, could generate the carbon and energy sources for the sulphate reducing bacteria at 25 degrees C. When comparing the results obtained at 25 degrees C with those obtained at 37 degrees C, it was observed that these were similar. However, at the higher temperature a faster flow-rate to the reactor was possible. The findings implied that the biological sulphate removal system described can be operated more economically at 25 degrees C than at 37 degrees C, as heating the reactor system can be omitted. PMID- 19809136 TI - Reducing microbial contamination in storm runoff from high use areas on California coastal dairies. AB - High use areas are a fundamental part of California coastal dairies and grazing livestock ranches as feeding areas, nurseries, and sick pens. High stocking densities and daily use in these areas lead to soil surfaces devoid of vegetation and covered in manure, with high potential for manure transport during winter rains to receiving waters regulated for shellfish harvesting and recreation. We characterized the association between California's Mediterranean climate and a series of existing and proposed management practices on fecal coliform bacteria (FCB) transport from high use areas on dairies and ranches. Results from 351 storm runoff samples collected below 35 high-use areas indicate that removal of cattle during winter, locating high use areas on level ground, application of straw and seeding, and vegetative buffer strip implementation were significantly associated with FCB concentration and load reductions. These results complement our findings for reductions of specific pathogens in runoff from these areas. These findings have practical significance because they document surface water quality benefits that the studied management practices provide in application on working farms and ranches. This direction is critical and timely for on-farm management efforts seeking to reduce microbial pollution in runoff and comply with indicator bacteria water quality criteria. PMID- 19809137 TI - Molecular assessment of salt-tolerant, perchlorate- and nitrate-reducing microbial cultures. AB - The microbial ecology of enrichment cultures adapted to the removal of perchlorate and nitrate from high salt solutions and ion-exchange brines was examined over a period of four years using denaturing gradient gel electrophoresis and total DNA extraction with cloning and in each case partial sequencing of the 16S rDNA genes. The cultures studied were a result of enrichment from marine sediment inoculum initiated in 2001. The resulting enrichment cultures were fed perchlorate, or perchlorate and nitrate, in a 3% (w/v) NaCl defined medium or ion-exchange brines (5.6% NaCl) containing perchlorate and nitrate with acetate as the electron donor. All of the sequences' closest matches in the NCBI GenBank database were to marine or salt-tolerant organisms. Strains belonging to the genera Halomonas or Marinobacter were found to dominate in cultures that were fed nitrate in addition to perchlorate, but were effectively absent from cultures fed perchlorate alone. The cultures fed perchlorate as the sole electron acceptor were relatively diverse with the dominant sequences belonging to the genera Dechloromarinus and Denitromonas. A study examining the effects of growing the cultures on different electron acceptors to the cultures revealed that Denitromonas may be more dominant than Dechloromarinus as the salt-tolerant, perchlorate-reducing organism. PMID- 19809138 TI - Decision support systems in water and wastewater treatment process selection and design: a review. AB - The continuously changing drivers of the water treatment industry, embodied by rigorous environmental and health regulations and the challenge of emerging contaminants, necessitates the development of decision support systems for the selection of appropriate treatment trains. This paper explores a systematic approach to developing decision support systems, which includes the analysis of the treatment problem(s), knowledge acquisition and representation, and the identification and evaluation of criteria controlling the selection of optimal treatment systems. The objective of this article is to review approaches and methods used in decision support systems developed to aid in the selection, sequencing of unit processes and design of drinking water, domestic wastewater, and industrial wastewater treatment systems. Not surprisingly, technical considerations were found to dominate the logic of the developed systems. Most of the existing decision-support tools employ heuristic knowledge. It has been determined that there is a need to develop integrated decision support systems that are generic, usable and consider a system analysis approach. PMID- 19809139 TI - Fractional analysis of arsenic in subsurface-flow constructed wetlands with different length to depth ratios. AB - Arsenic (As) removal in subsurface-flow constructed wetlands (CW) planting with vetiver grasses was experimented by comparing between two different configurations; (i) deep-bed units (dpCW) with length to depth (L:D) ratio=2 and (ii) shallow-bed units (shCW) with L:D ratio=8; operating at hydraulic retention time (HRT) of 6, 9, and 12 days. The tracer study of CW units revealed that no effect of L:D ratio on dispersion number could be determined, but affecting to the effective volume ratio. Based on the data obtained from the pilot-scale experiments of CW units for 117 days, it is apparent that the dpCW could achieve relatively high As removals (52.9%, 59.2%, and 72.1% at HRT of 6, 9, and 12 days, respectively). Analysis of As mass balance showed that only 0.2-0.4% of As input was uptaken by vetiver grasses whereas the major portion was retained in the CW media (38.9-77.6%). Forms of the retained As was determined by sequential fractionation which could indicate As complexation with iron and manganese on the media surface of 31-38% and As trapping into the media of 42-52% of the total. No obvious difference of As fractions in bed of between dpCW and shCW units was observable. PMID- 19809140 TI - Identification of physical parameters controlling the dominance of algal species in a subtropical reservoir. AB - Eutrophication is a serious problem of water resource management in Taiwan. The occurrence of annoying algal species as well as abnormally abundant algal mass threatens the quality of water supply. The growth and decline of a specific phytoplankton species are affected by environmental factors, including light, nutrients, temperature, etc. There have been many investigations on the effects of individual factors on the abundance and composition of algal populations. However, many analyses on the effects of environmental factors, especially the concentration of nutrients, on phytoplankton failed to identify the controlling factors on the dynamic change of the phytoplankton species. This study used statistical methods to isolate the effect of seasons on the phytoplankton growth and searched for the relationships between the nutrient concentrations and the abundance of different algal species in Feitsui Reservoir based on the data obtained from 1995 to 2003. We found that the dynamic change of dominance of some species of phytoplankton was strongly related to the seasonal factors. The controlling factors of the survival of an algal species were the settling and mobility of the phytoplankton, the mixing depth and the vertical mixing strength of the water bodies. According to our preliminary findings, the influence of physical factors, varying seasonally, outweighs the influence of nutrients on the algal species composition in Feitsui Reservoir in Taiwan. PMID- 19809141 TI - Pollutant load removal efficiency of pervious pavements: is clogging an issue? AB - Pervious pavements in car parks and driveways reduce the peak runoff rate and the quantity of runoff discharged into urban drains as well as improve the stormwater quality by trapping the sediments in the infiltrated water. The paper focuses on presenting results from the laboratory tests carried out to evaluate water quality improvements and effects of long-term decrease in infiltration rates with time due to sediments trapping (clogging) within the pavement pores. Clogging was not found to be a major factor affecting pervious pavement performance after simulating 17 years of stormwater quality samples. PMID- 19809142 TI - Fenton treatment efficacy for the purification of different kinds of wastewater. AB - The Fenton chemistry comprises both the classical Fenton reagent and its modification, so-called Fenton-like techniques, which have received great attention as a promising technology for wastewater treatment. In the present study real wastewater from different sources (leachate from oil shale semicoke landfill, pharmaceutical effluents from medical ointment production, municipal landfill leachate and wastewater originated from food-processing) were treated by means of Fenton/Fenton-like systems. The effectiveness of wastewater treatment was assessed by COD removal. Additionally, biodegradability improvement (BOD7/COD) and acute toxicity reduction of investigated wastewater samples were observed. The application of the Fenton chemistry to wastewater samples with different origin resulted generally in 70% or higher COD removal. Thus, the Fenton could be effectively applied both as a single treatment method and pre treatment step to improve subsequent biodegradability of wastewater effluents. PMID- 19809143 TI - Chemically enhanced primary treatment (CEPT) for removal of carbon and nutrients from municipal wastewater treatment plants: a case study of Shanghai. AB - With Chemically Enhanced Primary Treatment (CEPT) as the short-term process, the capacity of Bailonggang Wastewater Treatment Plant accounts for almost 25% of the total capacity of wastewater treatment in Shanghai, China. However, shortly after this plant was placed in operation in 2004, it was found that the effluent of CEPT couldn't meet the new national discharge criteria. Although the removal of phosphate is almost 80%, chemical oxygen demand (COD) and ammonia nitrogen (NH3 N) in the effluent is frequently found to exceed the standards. The primary goal of this research is to investigate the possibility of optimizing the CEPT to make it meet the discharge criteria before it is upgraded to a secondary treatment. An oxidant is adopted to remove NH3-N, and a high performance polyaluminum chloride (HP-PACl) is synthesized to enhance the removal of COD. It is found that HP-PACl improves the removal of COD, and the oxidant enhances NH3-N removal effectively. However, to meet the requirement of a newly implemented stricter discharge standard, it is necessary to upgrade this CEPT to a secondary treatment. The results of this study provide scientific evidence for the upgrade of the Bailonggang Wastewater Treatment Plant. PMID- 19809144 TI - The influence of supply and sewerage area characteristics on water and sewerage companies responses to the Water Framework Directive. AB - Using the example of raw water quality this paper examines the relationship between different spatial characteristics (geographical and physical properties) of Water and Sewerage Companies (WaSCs) supply and sewage areas and response to the Water Framework Directive. Results were obtained from thematic analysis and content analysis of 14 interviews with WaSCs representatives. Principal component analysis and cluster analysis of 51 WaSCs business function characteristics was employed to derive groups of similar WaSCs. Results indicate that there is difference in how WaSCs approach raw water quality issues. It appears that small WaSCs with relatively large agricultural areas in their supply catchments are more likely to seek managerial solutions to raw water quality problems. PMID- 19809145 TI - Estimation of lateral water flow and bromide transport in a subsurface seepage irrigation system. AB - Subsurface seepage irrigation is a common method used by growers in the Tri County Agricultural Area (TCAA), Florida, USA, owing to its cost-effectiveness and low maintenance requirements. This study investigated the lateral flow of the perched water and the lateral transport of bromide (Br-) in this irrigation system in the TCAA and estimated the potential discharge of Br- into the drainage canals at the edges of the field, using the Visual MODFLOW/ MT3DMS models in conjunction with field experiments. Simulations showed that the perched water flowed from the northeast to the southwest of the field. Migration of the Br- plume from the source areas toward the canals was very slow and varied depending on the selection of the outer Br- concentration contour levels. However, the lateral transport of Br- from the perched water into the canals occurred after about 61 days. The simulations further revealed that the rate of perched water Br discharge into the canals averaged 8.6 g day(-1) during a 30-day discharge period (from 61 to 91 days). This rate is very important for estimating Br- discharge into the canals and could also provide useful information for evaluating dissolved nutrient discharge into canals from the subsurface seepage irrigation system. PMID- 19809146 TI - Implications of reactor type and conditions on first-order hydrolysis rate assessment of maize silage. AB - The biodegradability and first-order hydrolysis coefficient of maize silage have been assessed from batch experiments using different types of inoculum and substrate to inocula (S/I) ratios, and from CSTRs working at different hydraulic retention times (HRTs). In the batch experiments, the assessed maximum biodegradability of the maize silage was 68 (+/-2.7)% and 73(+/-2.9)% while the first order hydrolysis was 0.26 (+/-0.01) and 0.27(+/-0.02) d(-1), using granular and a mixture of granular and suspended inoculum, respectively. In the CSTR experiment biodegradability ranged from 41-65% depending on the HRT applied whereas the calculated first order hydrolysis coefficient was 0.32 d(-1). It is concluded that batch experiments can be used to assess first order hydrolysis constants and biodegradability provided that a well balanced inoculum is guaranteed. Further, it is shown that CSTR reactors digesting maize silage and operating at HRTs as low as 20 days can attain 88% of maximum biodegradability as long as pH fluctuations are minimized. 2 mmol NaHCO3 per gram maize silage was calculated to suffice for the purpose. PMID- 19809147 TI - Detection and quantification of Microcystis spp. and microcystin-LR in Western Lake Erie during the summer of 2007. AB - Microcystis spp. blooms have occurred annually in western Lake Erie since about 1995. Microcystis produce a group of toxins known as microcystins which can be harmful to livestock and to humans. In this study, surface water samples were collected from six sites during six sampling events from July to October in 2007. In situ environmental data (e.g. pH, temperature) and laboratory analyses (e.g. nutrients) were carried out to characterize the six sites. The Microcystis spp. density ranged from 10(2) to 10(7) cells/ml. Microcystin-LR concentration of 20 of all 36 samples were below the detection limit (0.15-5 ppb), while the microcystin-LR concentration in the 16 remaining samples ranged from 0.5 to 3 x 10(3) microg per gram dry weight. The aim of this research was to investigate the relationships between sampling location, environmental parameters, Microcystis spp. concentration, and microcystin-LR concentration. The results suggest that temperature, nutrient concentration, turbidity, and wind speed and direction (P<0.05) are factors which affected Microcystis spp. density. Sampling site 8M, located 13 m from the Maumee River, provided an advantage for Microcystis spp. growth, presumably due to intermediate water depth (5.5 m) combined with impact from the river. No relationship was found between Microcystis spp. density and microcystin-LR concentration. Temperature, nutrient concentration and DO (P<0.05) were associated with the production of microcystin-LR. PMID- 19809148 TI - Dry and wet weather microbial characterization of the Chicago area waterway system. AB - The Chicago Area Waterway System (CAWS) is a man-made channel, which serves the Chicago area for the drainage of urban storm water and the conveyance of secondary treated effluent from the Metropolitan Water Reclamation District of Greater Chicago's (District) North Side, Stickney and Calumet water reclamation plants (WRPs). A microbial characterization of the CAWS upstream and downstream of the WRPs and from the WRP outfall was initiated by collecting dry and wet weather samples and analyzing for indicators and pathogens. During dry weather, indicator bacteria (fecal coliform [FC], E. coli [EC], enterococci [EN]) were the most abundant microbial species detected in the CAWS compared to pathogens (Salmonella spp [SA], enteric viruses [EV], adenovirus [AV], norovirus [NV] and Giardia and Cryptosporidium). Pseudomonas aeruginosa [PA] levels in the outfall samples were either lower or equivalent to the CAWS. The wet weather samples had a higher frequency of detection of indicator bacteria and pathogens compared to dry weather samples. Overall, the concentrations of pathogens in the CAWS, representing the weather conditions experienced in a recreational year, were relatively low. The study concluded that the presence of pathogens in the CAWS downstream of the WRPs were due to secondary loading of the waterway under wet weather conditions from combined sewer overflows (CSOs) and other discharges. PMID- 19809149 TI - A methodological approach for direct quantification of the activated sludge floc size distribution by using different techniques. AB - The activated sludge floc size distribution (FSD) is investigated by using different measurement techniques in order to gain insight in FSD assessment as well as to detect the strengths and limitations of each technique. A second objective was to determine the experimental conditions that allow a representative and accurate measurement of activated sludge floc size distributions. Laser diffraction, Time Of Transition (TOT) and Dynamic Image Analysis (DIA) devices were connected in series. The sample dilution liquid, the dilution factor and hydraulic flow conditions avoiding flocculation proved to be important. All methods had certain advantages and limitations. The MastersizerS has a broader dynamic size range and provides accurate results at high concentrations. However, it suffers from an imprecise evaluation of small size flocs and is susceptible to particle shape effects. TOT suffers less from size overestimation for non-spherical particles. However, care should be taken with the settings of the transparency check. Being primarily a counting technique, DIA suffers from a limited size detection range but is an excellent technique for process visualization. All evaluated techniques turned out to be reliable methods to quantify the floc size distribution. Selection of a certain method depends on the purpose of the measurement. PMID- 19809150 TI - Tertiary treatment of slaughterhouse effluent: degradation kinetics applying UV radiation or H2O2/UV. AB - In some Brazilian regions, surface water has become scarce, e.g. semi arid climate areas and densely populated and industrial areas, where water over exploitation and/or fluvial pollution has been more common. Advanced oxidative processes (AOP) provide treated water as a source of reuse water even with the characteristics of drinking water enabling water reuse practices also in food industries. The secondary wastewater of a slaughterhouse was the water source for a tertiary treatment study evaluating the kinetics of the photo-induced degradation of color and UV254 under UV radiation with and without the addition of H2O2. The proximity of the k' values of color and UV254 degradation by UV indicates that the compounds responsible for color may be the same content measured by UV254. The H2O2/UV treatment was 5.2 times faster than simple UV in removing aromatic compounds. The degradation kinetics of aromatic compounds in both treatments followed a pseudo-first order law. The pseudo-first order constant for H2O2/UV and UV treatments were kUV254'=0.0306 min(-1) and kUV254'=0.0056 min(-1), respectively. PMID- 19809151 TI - Model-based comparison of two ways to enhance WWTP capacity under stormwater conditions. AB - Two different approaches to increase the fraction of combined water treated in the wastewater treatment plant (WWTP) which would otherwise contribute to combined sewer overflows (CSO) are presented and compared based on modelling results with regard to their efficiencies during various rain events. The first option is to generally increase the WWTP inflow according to its actual capacity rather than pre-setting a maximum that applies to worst case loading. In the second option the WWTP inflow is also increased, however, the extra inflow of combined water is bypassing the activated sludge tank and directly discharged to the secondary clarifier. Both approaches have their advantages. For the simulated time series with various rain events, the reduction of total COD load from CSOs and WWTP effluent discharged to the receiving water was up to 20% for both approaches. The total ammonia load reduction was between 6% for the bypass and 11% for inflow increase. A combination of both approaches minimises the adverse effects and the overall emission to the receiving water. PMID- 19809152 TI - Degradation of pesticide residues in vineyard effluents by activated sludge treatment. AB - The study evaluates the feasibility of the degradation of thirteen pesticides by an activated sludge aerobic treatment, used for the purification of wastewater effluents produced from a winemaking process. Chemical analyses were used to determine the capacity of the treatment to eliminate the implied pollution hazard. The results show that the biological treatment of such pesticides by activated sludge is possible if a minimum adaptation period of approximately 8 days is applied. System stability is reached in 30 days. The related degradation of twelve pesticides was 97%, the exception being anilinopyrimidin. However, the sludge biomass is dispersed (the pesticides destroyed the biomass flocs) and it was necessary to add a flocculation stage to concentrate and remove the free micro-organisms by using sedimentation. PMID- 19809153 TI - Removing textile mill effluent recalcitrant COD and toxicity using the H2O2/UV system. AB - The potential of the H2O2/UV process for improving quality of an industrial textile effluent before biological treatment was evaluated in the laboratory using a multivariate experimental design to determine the effects of pH, H2O2 dose and reaction time on colour, COD and toxicity removal efficiencies. Increasing reaction time (from 10 to 120 min) and H2O2 dose (from 0 to 5 mmol L( 1)) significantly improved removal efficiencies, while increasing pH (from 4 to 10) had a negative effect on colour and toxicity removals. Laboratory H2O2/UV treatment of the mill effluent under optimum conditions (pH 7, 5 mmol L(-1) H2O2, 120 min) resulted in decreases in colour (70%), COD (21%) and toxicity (67%), without lowering BOD. H2O2 was consumed within the first 30-60 min, while the effluent average oxidation state stabilized after 60 min. Decreasing reaction time to 60 min resulted in similar colour (63%) and COD (20%) removals but lower toxicity removal (44%). Spectrophotometric monitoring of the optimized reaction indicated partial destruction of residual aromatic azo dyes. H2O2 and residual peroxide and average oxidation state of the effluent Effluent biodegradability (BOD/COD) increased by 28% after the H2O2/UV treatment. Improvements observed in effluent quality are expected to enhance combined AOP-biological treatment efficiency of the mill effluent. PMID- 19809154 TI - Optimization of intracellular microcystin-LR extraction for its analysis by protein phosphatase inhibition assay. AB - Microcystins are toxins produced by some strains of cyanobacteria. Several methods have been developed to allow the quantification of microcystins, which are mainly endotoxins. Among those methods, the protein phosphatase inhibition assay is a good candidate as a screening method because of its sensitivity, simplicity and specificity. In this work a method for intracellular microcystin extraction in field water samples and lab cyanobacterial cultures prior to their analysis by protein phosphatase inhibition assay has been optimized. Microcystin LR and Microcystis aeruginosa PCC 7806 were used as reference microcystin and strain, respectively, in order to optimize the protocol. The protocol consists on filtering the sample through a nylon filter of 0.8 microm, filter extraction with methanol 80% 0.1% trifluoroacetic acid (TFA) 0.1% tween 20, extract centrifugation and supernatant dilution (1/20). The establishment of an extraction protocol was carried out determining the extraction volume, time of extraction and number of extractions. The advantages of the method developed in this work are basically its simplicity and avoiding the use of specific and expensive equipment. PMID- 19809155 TI - Three dimensional modelling of interaction between surface and Darcy flow regimes through soils. AB - The present paper deals with the impact of surface flow on hydrodynamic conditions in saturated underground domains. A three dimensional finite element scheme has been used to simulate underground flow resulting from the flow of water over a saturated land. The results clearly show the effects of the surface flow on the hydrodynamic conditions of the subsurface porous regions. This analysis is an important prerequisite for the prediction of contaminant mobility in soils and hence provides a convenient tool for the prediction of interaction between surface and subsurface flow processes. For low permeability cases, considered here, the governing equations consist of water continuity and Darcy equations. These equations are solved using a robust and reliable finite element procedure. PMID- 19809156 TI - Influence and modelling of urban runoff on the peak flows in rivers. AB - Surface waters and urban drainage systems are usually studied separately. However there are important interactions between both systems. Urban drainage systems can have an important impact on the surface waters, mainly at combined sewer overflows. On the other hand during periods of high water levels in a river, the runoff from the urban drainage system can be significantly influenced by backwater, which increases the probability of flooding in is not obvious, because the modelling tools for both systems are often hard to combine properly. To properly assess the probability of flooding for this kind of integrated water systems, different submodels are needed for both subsystems. In practice often one single model is used to describe the runoff to rivers despite the presence of urban catchments. The main objective of this study is to show the limits of this simplified approach. Furthermore, it is necessary to use continuous long term simulations, because of the differences in runoff behaviour. Detailed hydrodynamic models do not really fit for this purpose because of long simulation times and high demands in memory and disk space. Therefore simplified conceptual models are more useful. PMID- 19809157 TI - Antigen-specific Tregs control T cell responses against a limited repertoire of tumor antigens in patients with colorectal carcinoma. AB - Spontaneous antitumor T cell responses in cancer patients are strongly controlled by Tregs, and increased numbers of tumor-infiltrating Tregs correlate with reduced survival. However, the tumor antigens recognized by Tregs in cancer patients and the impact of these cells on tumor-specific T cell responses have not been systematically characterized. Here we used a broad panel of long synthetic peptides of defined tumor antigens and normal tissue antigens to exploit a newly developed method to identify and compare ex vivo the antigen specificities of Tregs with those of effector/memory T cells in peripheral blood of colorectal cancer patients and healthy subjects. Tregs in tumor patients were highly specific for a distinct set of only a few tumor antigens, suggesting that Tregs exert T cell suppression in an antigen-selective manner. Tumor-specific effector T cells were detectable in the majority of colorectal cancer patients but not in healthy individuals. We detected differences in the repertoires of antigens recognized by Tregs and effector/memory T cells in the majority of colorectal cancer patients. In addition, only effector/memory T cell responses against antigens recognized by Tregs strongly increased after Treg depletion. The selection of antigens according to preexisting T cell responses may improve the efficacy of future immunotherapies for cancer and autoimmune disease. PMID- 19809158 TI - Semaphorin 3A is an endogenous angiogenesis inhibitor that blocks tumor growth and normalizes tumor vasculature in transgenic mouse models. AB - Tumor growth and progression rely upon angiogenesis, which is regulated by pro- and antiangiogenic factors, including members of the semaphorin family. By analyzing 3 different mouse models of multistep carcinogenesis, we show here that during angiogenesis, semaphorin 3A (Sema3A) is expressed in ECs, where it serves as an endogenous inhibitor of angiogenesis that is present in premalignant lesions and lost during tumor progression. Pharmacologic inhibition of endogenous Sema3A during the angiogenic switch, the point when pretumoral lesions initiate an angiogenic phase that persists throughout tumor growth, enhanced angiogenesis and accelerated tumor progression. By contrast, when, during the later stages of carcinogenesis following endogenous Sema3A downmodulation, Sema3A was ectopically reintroduced into islet cell tumors by somatic gene transfer, successive waves of apoptosis ensued, first in ECs and then in tumor cells, resulting in reduced vascular density and branching and inhibition of tumor growth and substantially extended survival. Further, long-term reexpression of Sema3A markedly improved pericyte coverage of tumor blood vessels, something that is thought to be a key property of tumor vessel normalization, and restored tissue normoxia. We conclude, therefore, that Sema3A is an endogenous and effective antiangiogenic agent that stably normalizes the tumor vasculature. PMID- 19809159 TI - Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models. AB - Rhabdomyosarcoma (RMS) is a childhood cancer originating from skeletal muscle, and patient survival is poor in the presence of metastatic disease. Few determinants that regulate metastasis development have been identified. The receptor tyrosine kinase FGFR4 is highly expressed in RMS tissue, suggesting a role in tumorigenesis, although its functional importance has not been defined. Here, we report the identification of mutations in FGFR4 in human RMS tumors that lead to its activation and present evidence that it functions as an oncogene in RMS. Higher FGFR4 expression in RMS tumors was associated with advanced-stage cancer and poor survival, while FGFR4 knockdown in a human RMS cell line reduced tumor growth and experimental lung metastases when the cells were transplanted into mice. Moreover, 6 FGFR4 tyrosine kinase domain mutations were found among 7 of 94 (7.5%) primary human RMS tumors. The mutants K535 and E550 increased autophosphorylation, Stat3 signaling, tumor proliferation, and metastatic potential when expressed in a murine RMS cell line. These mutants also transformed NIH 3T3 cells and led to an enhanced metastatic phenotype. Finally, murine RMS cell lines expressing the K535 and E550 FGFR4 mutants were substantially more susceptible to apoptosis in the presence of a pharmacologic FGFR inhibitor than the control cell lines expressing the empty vector or wild type FGFR4. Together, our results demonstrate that mutationally activated FGFR4 acts as an oncogene, and these are what we believe to be the first known mutations in a receptor tyrosine kinase in RMS. These findings support the potential therapeutic targeting of FGFR4 in RMS. PMID- 19809160 TI - Platelet depletion and aspirin treatment protect mice in a two-event model of transfusion-related acute lung injury. AB - Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion associated mortality in the US. Previously, we established an immune-mediated TRALI mouse model, wherein mice with cognate antigen were challenged with MHC class I mAb. In this study, when mice housed in a rodent, specific pathogen-free barrier room were challenged with MHC I mAb, there was significant protection from TRALI compared with nonbarrier mice. Priming mice with LPS restored lung injury with mAb challenge. Using TLR4-deficient bone marrow chimeras, the priming phenotype was restricted to animals with WT hematopoietic cells, and depletion of either neutrophils or platelets was protective. Both neutrophils and platelets were sequestered in the lungs of mice with TRALI, and retention of platelets was neutrophil dependent. Interestingly, treatment with aspirin prevented lung injury and mortality, but blocking the P selectin or CD11b/CD18 pathways did not. These data suggest a 2-step mechanism of TRALI: priming of hematopoietic cells, followed by vascular deposition of activated neutrophils and platelets that then mediate the severe lung injury. Furthermore, our data offer an explanation for the increased incidence of TRALI in patients with immune priming conditions, and we suggest what we believe to be a novel therapeutic approach. PMID- 19809161 TI - Selective modulation of TLR4-activated inflammatory responses by altered iron homeostasis in mice. AB - Mice deficient in the hemochromatosis gene, Hfe, have attenuated inflammatory responses to Salmonella infection associated with decreased macrophage TNF-alpha and IL-6 biosynthesis after exposure to LPS. In this study, we show that the abnormal cytokine production is related to impaired TLR4 signaling. Despite their abnormal response to LPS, Hfe KO macrophages produced amounts of TNF-alpha similar to those in WT cells after TLR2 stimulation. Consistent with this finding, LPS-induced activation of Mal/MyD88-dependent events was normal in the mutant macrophages. However, LPS-induced IFN-beta expression, a TRAM/TRIF dependent response activated by TLR4, was reduced by Hfe deficiency. This reduction could be replicated in WT macrophages with the use of iron chelators. In contrast, TLR3-activated expression of IFN-beta, a TRIF-dependent response, was normal in Hfe KO macrophages and was unaffected by iron chelation. Our data suggest that low intracellular iron selectively impairs signaling via the TLR4/TRAM/TRIF pathway proximal to TRIF and results in reduced LPS-induced cytokine expression. Furthermore, by mimicking the altered iron metabolism associated with Hfe deficiency, we found that 3 different inhibitors of hepcidin attenuated Salmonella-induced and noninfectious enterocolitis. Thus, manipulation of iron homeostasis could represent a new therapeutic approach to controlling inflammation. PMID- 19809162 TI - The glial cell response is an essential component of hypoxia-induced erythropoiesis in mice. AB - A key adaptation to environmental hypoxia is an increase in erythropoiesis, driven by the hormone erythropoietin (EPO) through what is traditionally thought to be primarily a renal response. However, both neurons and astrocytes (the largest subpopulation of glial cells in the CNS) also express EPO following ischemic injury, and this response is known to ameliorate damage to the brain. To investigate the role of glial cells as a component of the systemic response to hypoxia, we created astrocyte-specific deletions of the murine genes encoding the hypoxia-inducible transcription factors HIF-1alpha and HIF-2alpha and their negative regulator von Hippel-Lindau (VHL) as well as astrocyte-specific deletion of the HIF target gene Vegf. We found that loss of the hypoxic response in astrocytes does not cause anemia in mice but is necessary for approximately 50% of the acute erythropoietic response to hypoxic stress. In accord with this, erythroid progenitor cells and reticulocytes were substantially reduced in number in mice lacking HIF function in astrocytes following hypoxic stress. Thus, we have demonstrated that the glial component of the CNS is an essential component of hypoxia-induced erythropoiesis. PMID- 19809163 TI - The tetramer structure of the glycoside hydrolase family 27 alpha-galactosidase I from Umbelopsis vinacea. AB - The crystal structure of Umbelopsis vinacea alpha-galactosidase I, which belongs to glycoside hydrolase family 27, was determined at 2.0 A resolution. The monomer structure was well conserved with those of glycoside hydrolase family 27 enzymes. The biological tetramer structure of this enzyme was constructed by the crystallographic 4-fold symmetry, and tetramerization appeared to be caused by three inserted peptides that were involved in the tetramer interface. The quaternary structure indicated that the substrate specificity of this enzyme might be related to the tetramer formation. Three N-glycosylated sugar chains were observed, and their structures were found to be of the high-mannose type. PMID- 19809164 TI - Different suppressive effects of fucoidan and lentinan on IL-8 mRNA expression in in vitro gut inflammation. AB - A system for assessing the anti-inflammatory effects of food factors was developed by establishing a co-culture system with intestinal epithelial Caco-2 cells and macrophage RAW264.7 cells. The results indicate that fucoidan and lentinan exhibited different suppressive effects on interleukin-8 (IL-8) mRNA expression in Caco-2 through tumor necrosis factor-alpha (TNF-alpha) production from RAW264.7 stimulated with lipopolysaccharide (LPS). PMID- 19809165 TI - Luteolin increases the expression of GRP94 in HeLa cells. AB - We found increased expression of GRP94, the 94-kDa glucose-regulated protein, in HeLa cells 24 h after treatment with luteolin. Luteolin increased the levels of GRP94 mRNA and protein, but it did not increase the expression of unfolded protein response (UPR)-regulated genes. In addition, luteolin also enhanced GRP94 promoter activity, suggesting that it enhances the expression of GRP94 at the transcriptional level, not via the UPR signaling pathway. PMID- 19809166 TI - Production of glycolipid biosurfactants, mannosylerythritol lipids, using sucrose by fungal and yeast strains, and their interfacial properties. AB - Glycolipid biosurfactants, mannosylerythritol lipids (MELs), were produced from glucose and sucrose without vegetable oils. Pseudozyma antarctica JCM 10317, Ustilago maydis NBRC 5346, U. scitaminea NBRC 32730, and P. siamensis CBS 9960 produced mainly MEL-A, MEL-A, MEL-B, and MEL-C respectively. The sucrose-derived MELs showed excellent interfacial properties: low critical micelle concentration as well as that of oil-derived MELs. PMID- 19809167 TI - Inhibitive effects of alkyl gallates on hyaluronidase and collagenase. AB - A series of the gallate esters of n-alkanols (C(1)-C(12)) was examined to determine their inhibitory activities against hyaluronidase and collagenase. Hexyl, heptyl, octyl, nonyl, and decyl gallates inhibited both hyaluronidase and collagenase, and the most potent inhibitor was octyl gallate against both enzymes. Octyl 3,5-dihydroxybenzoate showed inhibitory effects on hyaluronidase, whereas collagenase was inhibited by octyl 3,4-dihydroxybenzoate. PMID- 19809168 TI - Proline and glycinebetaine confer cadmium tolerance on tobacco bright yellow-2 cells by increasing ascorbate-glutathione cycle enzyme activities. AB - Cadmium (Cd) stress significantly decreased membrane integrity and impaired the ascorbate (ASC)-glutathione (GSH) cycle in tobacco Bright Yellow-2 cells. Exogenous application of proline and glycinebetaine (betaine) significantly restored the membrane integrity and increased the activities of ASC-GSH cycle enzymes under Cd stress without maintenance of the rich ASC or GSH pools. Moreover, proline offered more efficient protection against Cd stress than betaine. PMID- 19809169 TI - Amino acid substitutions and intragenic duplications of Bacillus sp. PS3 flagellin cause complementation of the Bacillus subtilis flagellin deletion mutant. AB - Bacillus sp. PS3 produces a glycosylated flagellin. In this study, a number of the glycosylated residues of the flagellin protein were found to be located in the central variable region of this protein. We also report that the motility defect of the Bacillus subtilis flagellin mutant was complemented by Bacillus sp. PS3 flagellin variants without glycosylation, which contained amino acid substitutions and intragenic duplications in the variable region of flagellin. PMID- 19809170 TI - Quantitative analysis of the effects of diffusates from plant roots on the hatching of Meloidogyne chitwoodi from young and senescing host plants. AB - We analyzed the effects of diffusates from six different crops on the egg hatching of Meloidogyne chitwoodi collected from young and senescing tomato plants using a non-linear model. Diffusates only from tomato and carrot significantly delayed the hatching of eggs from young plants, but did not do so from senescing plants, suggesting that delay of hatching depends on the origin of diffusates and host plant age. PMID- 19809171 TI - Anti-obesity effect on rodents of the traditional Japanese food, tororokombu, shaved Laminaria. AB - Tororokombu is a traditional Japanese food made from edible kelp. The way to make tororokombu is characterized by shaving kelp very thinly. It was found that tororokombu decreased the serum triglyceride level induced by oil administration to rats and had an anti-obesity effect on obese mice induced by a high-fat diet. These effects were more powerful than those of non-shaved kelp. PMID- 19809172 TI - Antihypertensive effect of enzymatic hydrolysate of collagen and Gly-Pro in spontaneously hypertensive rats. AB - Continuous oral feeding of enzymatic hydrolysate of porcine skin collagen showed an antihypertensive effect in spontaneously hypertensive rats (SHRs). We isolated an angiotensin I-converting enzyme (ACE) inhibitory peptide, Gly-Phe-Hyp-Gly-Pro (IC(50)=91 microM), from the hydrolysate, but the ACE inhibitory activities of the other peptides isolated were weak. Although the ACE inhibitory activity of Gly-Pro (IC(50)=360 microM) was not potent, Gly-Pro exists in collagen as a large number of repeated sequences. We then examined the antihypertensive effect of Gly Pro. Orally administered Gly-Pro at 500 mg/kg significantly decreased the blood pressure of SHRs, and at 50 mg/kg it also showed a tendency to lower the blood pressure. Oral administration of Gly-Phe-Hyp-Gly-Pro (10 or 30 mg/kg) also decreased the blood pressure of SHRs. PMID- 19809173 TI - Cathepsin D-like aspartic proteinase occurring in a maize weevil, Sitophilus zeamais, as a candidate digestive enzyme. AB - The maize weevil, Sitophilus zeamais, is an insect pest infesting rice and corn seeds. We identified an aspartic proteinase (AP) digesting rice glutelin in the alimentary tract of S. zeamais. The mRNA encoding the AP (SAP1) was expressed in the larvae foregut and in the adult midgut. These results indicate that SAP1 is probably digestive enzyme of S. zeamais. PMID- 19809174 TI - Anthocyanin components and mechanism for color development in blue Veronica flowers. AB - 3-Di-p-coumaroylsophoroside-5-malonylglucoside and its demalonyl derivative were isolated from blue petals of Veronica persica Poiret. Blue, violet and purple cells coexist in the petal. These colors might be due to the varying pH of the vacuole between 5 and 7 unit. Only the demalonylated pigment was detected in the blue anthers. PMID- 19809175 TI - Stereochemistry of female-specific normonoterpenes, sex pheromone candidates from the acarid mite, Tyreophagus sp. (Astigmata: Acaridae). AB - Two normonoterpenes were detected from an unidentified Tyreophagus sp. as new female-specific components. Both planar structures were identified to be 2,6 dimethyl-5-heptenal (1) and 2,6-dimethyl-5-hepten-1-ol (2) by GC/MS co chromatography with synthetic 1 and 2. The stereochemistry of 2 was determined to be R by a GC analysis with a chiral column, while that of 1 was presumed to be similar to 2 based on the biosynthetic aspects. PMID- 19809176 TI - Mustard oil in "Shibori Daikon" a variety of Japanese radish, selectively inhibits the proliferation of H-ras-transformed 3Y1 cells. AB - Cruciferous vegetables and their isothiocyanates are promising foods and agents for cancer prevention. We focus here on the effects of mustard oil (SMO) in a variety of the Japanese radish, Shibori Daikon (Raphanus sativus), on the proliferation of 3Y1 rat fibroblasts and the H-ras-transformed derivative, HR-3Y1 2. SMO (1.6 microg/ml) inhibited the proliferation of HR-3Y1-2, but not 3Y1 after 24 h after treatment. A cell cycle analysis showed that SMO induced G2/M arrest after 6 h, although this effect was not observed 24 h after the treatment. SMO transiently decreased the cellular reduced glutathione level accompanied with up regulation of the intracellular reactive oxygen species 2-3 h post-treatment. Glutathione ethyl ester and N-acetyl-L-cysteine prevented the growth inhibitory effect of SMO. This mustard oil extract consisted of 95.6% 4-methylthio-3-butenyl isothiocyanate and 4.4% 4-methylthiobutyl isothiocyanate. SMO selectively inhibited H-ras-transformed 3Y1 cells associated with transient oxidative stress via reduced glutathione (GSH) depletion. PMID- 19809177 TI - Analysis of hydrogen peroxide in an aqueous extract of cigarette smoke and effect of pH on the yield. AB - An analysis of hydrogen peroxide in an aqueous extract of cigarette smoke, which contains many redox-active compounds, requires a method with high selectivity. An aqueous extract of the particulate phase of cigarette smoke was analyzed by HPLC with an electrochemical detector (ECD). Samples were prepared by collecting the particulate phase of the cigarette smoke on a glass fiber filter and extracting it with a phosphate buffer. The obtained solution was purified by using a Waters Oasis MCX cation-exchange cartridge, and then analyzed by an HPLC-ECD system with a Shodex KS-801 mixed-mode resin column. Pre-injecting hydrogen peroxide at a high concentration into the HPLC instrument stabilized the analytical results. The recovery of hydrogen peroxide by using an extract of the particulate phase of the cigarette smoke was more than 80%. An increase in the amount of hydrogen peroxide was observed during extraction with the phosphate buffer at higher pH values. In contrast, extraction with phosphoric acid did not increase the amount of hydrogen peroxide during extraction. PMID- 19809178 TI - Sardine peptide with angiotensin I-converting enzyme inhibitory activity improves glucose tolerance in stroke-prone spontaneously hypertensive rats. AB - An enzymatic hydrolysate of sardine protein (sardine peptide, SP) derived from sardine muscle possesses angiotensin I-converting enzyme (ACE) inhibitory activity. In the present study, we investigated the effect of SP on the blood glucose levels in stroke-prone spontaneously hypertensive rats (SHRSPs). Ten-week old SHRSPs were assigned to three groups. The control group was given tap water for 4 weeks, while the experimental groups were given water containing SP (1 g/kg/d) or an ACE inhibitor, captopril (8 mg/kg/d). Treatment with SP and captopril decreased ACE activity in the kidney, aorta, and mesentery. There were no differences in fasting blood glucose levels among the three groups, whereas SP and captopril administration significantly suppressed the increase in blood glucose after glucose loading in the control SHRSPs. No difference was observed in plasma insulin levels among the three groups. Thus treatment with captopril and ACE-inhibitory sardine peptides ameliorated the glucose tolerance of this rat strain. PMID- 19809179 TI - Apple procyanidins induced vascular relaxation in isolated rat aorta through NO/cGMP pathway in combination with hyperpolarization by multiple K+ channel activations. AB - Apple (Malus pumila) procyanidins led to a potent vasorelaxation effect in 1.0 microM phenylephrine-contractive rat thoracic aorta. Relaxation was greatly reduced by 70 mM KCl as well as by removal of the endothelium, suggesting that it was associated with endothelium-dependent hyperpolarization. Neither cAMP synthesis inhibition nor NAD(P)H oxidase inhibition abolished the effect. In contrast, complete abolition by a soluble guanylyl cyclase inhibitor revealed that apple procyanidins were mainly involved in the cGMP production pathways. In the presence of N(G)-monoethyl-L-arginine or tetraethylammonium chloride, the effect was still observed at higher concentrations (>25 microg/ml), while their combination completely diminished the effect. Vasorelaxation was to some extent affected by paxillin, apamin and glybenclamide, and was greatly affected by 4 aminopyridine and by BaCl(2). These results indicate that procyanidin-induced vasorelaxation is associated with NO-cGMP pathway in combination with hyperpolarization due to multiple activation of Ca(2+)-dependent and -independent K(+) channels. PMID- 19809180 TI - Development of a novel functional high-throughput screening system for pathogen effectors in the yeast Saccharomyces cerevisiae. AB - Our understanding of the molecular mechanisms of bacterial pathogenesis has been improved especially by the discovery of host cell contact-dependent secretion systems such as the type-III secretion system (T3SS) found in numerous pathogens. Although the identification of pathogen effectors translocated into host cells through T3SS is essential to the understanding of pathogenesis, their general sequence uniqueness confound attempts to identify such proteins by sequence homology. Here we report the development of a functional high-throughput screening system for pathogen effectors in yeast that consists of a Gateway(TM) compatible Tet-Off inducible expression vector and a yeast strain expressing a reporter, facilitating identification of the effectors affecting host vesicular trafficking pathways. We evaluated this system and optimized the screening condition using several known pathogen effectors. We found this system useful in functional characterization of pathogen effector and it can be adapted to functional high-throughput screening as well. PMID- 19809181 TI - Immobilization of Bacillus licheniformis L-arabinose isomerase for semi continuous L-ribulose production. AB - Bacillus licheniformis L-arabinose isomerase (BLAI) with a broad pH range, high substrate specificity, and high catalytic efficiency for L-arabinose was immobilized on various supports. Eupergit C, activated-carboxymethylcellulose, CNBr-activated agarose, chitosan, and alginate were tested as supports, and Eupergit C was selected as the most effective. After determination of the optimum enzyme concentration, the effects of pH and temperature were investigated using a response surface methodology. The immobilized BLAI enzyme retained 86.4% of the activity of the free enzyme. The optimal pH for the immobilized BLAI was 8.0, and immobilization improved the optimal temperature from 50 degrees C (free enzyme) to a range between 55 and 65 degrees C. The half life improved from 2 at 50 degrees C to 212 h at 55 degrees C following immobilization. The immobilized BLAI was used for semi-continuous production of L-ribulose. After 8 batch cycles, 95.1% of the BLAI activity was retained. This simple immobilization procedure and the high stability of the final immobilized BLAI on Eupergit C provide a promising solution for large-scale production of L-ribulose from an inexpensive L arabinose precursor. PMID- 19809182 TI - Autophagy inhibition enhances apoptosis induced by ginsenoside Rk1 in hepatocellular carcinoma cells. AB - Our previous study indicated that ginsenoside Rk1 has anti-tumor activity and that its mode of action in HepG2 cells treated for 48 h involves coordinated inhibition of telomerase and induction of apoptosis. In the present study, we found that Rk1 induces both G(1) phase arrest and autophagy, but not apoptosis, at an earlier stage of treatment. A 24-h incubation of HepG2cells with Rk1 induced G(1) phase arrest. Rk1-induced autophagy was documented by the conversion of microtubule associated protein light chain 3 (LC3)-I to LC3-II, an autophagosome marker, and monodansylcadaverine (MDC) incorporation into autolysosomes. Combination of Rk1 with an autophagy inhibitor, such as bafilomycin A1 or beclin 1 siRNA, enhanced the anti-tumor effect of Rk1. These results imply that autophagy functions as a survival mechanism in HepG2 cells against Rk1-induced apoptosis. Taken together, our results support the use of autophagy inhibitors in combination with Rk1 as an effective anti-cancer regimen in HepG2 cells. PMID- 19809183 TI - Structures and properties of a diastereoisomeric molecular compound of (2S,3S)- and (2R,3S)-N-acetyl-2-amino-3-methylpentanoic acids. AB - An X-ray crystal structural analysis revealed that (2S,3S)-N-acetyl-2-amino-3 methylpentanoic acid (N-acetyl-L-isoleucine; Ac-L-Ile) and (2R,3S)-N-acetyl-2 amino-3-methylpentanoic acid (N-acetyl-D-alloisoleucine; Ac-D-aIle) formed a molecular compound containing one Ac-L-Ile molecule and one Ac-D-aIle molecule as an unsymmetrical unit. This molecular compound is packed with strong hydrogen bonds forming homogeneous chains consisting of Ac-L-Ile molecules or Ac-D-aIle molecules and weak hydrogen bonds connecting these homogeneous chains in a fashion similar to that observed for Ac-L-Ile and Ac-D-aIle. Recrystallization of an approximately 1:1 mixture of Ac-L-Ile and Ac-D-aIle from water gave an equimolar molecular compound due to its lower solubility than that of Ac-D-aIle or especially Ac-L-Ile. The results suggest that the equimolar mixture of Ac-L Ile and Ac-D-aIle could be obtained from an Ac-L-Ile-excess mixture by recystallization from water. PMID- 19809184 TI - Properties of ethanol fermentation by Flammulina velutipes. AB - Basidiomycetes have the ability to degrade lignocellulosic biomass, and some basidiomycetes produce alcohol dehydrogenase. These characteristics may be useful in the direct production of ethanol from lignocellulose. Ethanol fermentation by basidiomycetes was investigated to examine the possibility of ethanol production by consolidated bioprocessing (CBP) using Flammulina velutipes. F. velutipes converted D-glucose to ethanol with a high efficiency (a theoretical ethanol recovery rate of 88%), but ethanol production from pentose was not observed. These properties of F. velutipes are similar to those of Saccharomyces cerevisiae, but the basidiomycete converted not only sucrose, but also maltose, cellobiose, cellotriose, and cellotetraose to ethanol, with almost the same efficiency as that for D-glucose. From these results, we concluded that F. velutipes possesses advantageous characteristics for use in CBP. PMID- 19809185 TI - Synthesis of the deuterated sex pheromone components of the grape borer, Xylotrechus pyrrhoderus. AB - Adult males of the grape borer, Xylotrechus pyrrhoderus, secrete (S)-2-hydroxy-3 octanone [(S)-1] and (2S,3S)-2,3-octanediol [(2S,3S)-2] from their nota of prothoraces as sex pheromone components. Their structural similarity suggests that one of them is the biosynthetic precursor of the other component. In order to confirm the biochemical conversion, deuterated derivatives of both components were synthesized by starting from a Wittig reaction between hexanal and an ylide derived from D(5)-iodoethane and ending with enantiomeric resolution by chiral HPLC. The molecular ions of 1 and 2 could scarcely be detected by using a GC-MS analysis, and the labeled compounds showed similar mass spectra to the unlabeled pheromone components. However, several fragment ions, including four deuterium atoms, were observed in the mass spectra of their acetate derivatives, indicating that the conversion could be confirmed by examining a compound with the diagnostic ions after acetylation of the volatiles collected from insects treated with the labeled precursors. PMID- 19809186 TI - Inhibitory effect of phytoglycoprotein (115 kDa) on the expression of TNF-alpha and interleukin-1beta via inhibition of MAP kinase in primary cultured mouse thymocytes. AB - The present study was performed to investigate the anti-inflammatory potential of 115 kDa glycoprotein isolated from Zanthoxylum piperitum DC leaves (ZPDC glycoprotein) in primary cultured mouse thymocytes. To determine whether the ZPDC glycoprotein has inhibitory capacity against inflammation in vitro, we evaluated the activities of inflammation-related factors such as phosphorylations of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK) 1/2, and the activities of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) in 12-O-tetradecanoylphorbol 13-acetata (PMA, 50 nM)-treated mouse thymocytes. Our results showed that the ZPDC glycoprotein (200 mug/ml) has a suppressive effect on the expression of MAPK (ERK1/2 and p38 MAPK), on mRNA expression of pro inflammatory cytokines (TNF-alpha and IL-1beta), and on protein expression of pro inflammatory proteins (iNOS and COX-2). We speculate that the ZPDC glycoprotein is an example of a natural compound that blocks pro-inflammatory signal transduction pathways. PMID- 19809187 TI - First synthesis of (+/-)-basidifferquinone C, an inducer for fruiting-body formation in Polyporus arcularius. AB - Basidifferquinones, isolated from Streptomyces sp., are potent inducers of fruiting-body formation in the basidiomycete, Polyporus arcularius. The first synthesis of (+/-)-basidifferquinone C was accomplished by starting from 3,5 dihydroxy-2-naphthoic acid. PMID- 19809188 TI - Crystal structures of sodium-bound annexin A4. AB - Annexin A4 (Anx4) possesses four repeat domains with one Ca(2+)-binding site (CBS) in each domain. In this study, we resolved two crystal structures of the Na(+)-bound form at high resolution (1.58 and 1.35 A). This is the first report that Anx4 binds the Na(+) ion in CBSs. Electron density maps, valence screening, and atomic absorbance spectrometry confirmed that Anx4 bound the Na(+) ion. One structure (1.58 A) bound the Na(+) ion in CBS I, whereas another structure (1.35 A) bound the Na(+) ion in CBS II and CBS III. We compared the two Na(+)-bound forms by superimposing their C(alpha) traces. The C(alpha) atoms of CBS III largely moved by coordination of the Na(+) ion. In the C(alpha) atoms of CBS I, however, little change resulted from Na(+)-coordination. Only the side chain of Glu71 was moved by Na(+)-coordination in CBS I. These results indicate that Anx4 also binds not only Ca(2+) but also Na(+) ion in the CBS. PMID- 19809189 TI - Synthesis of benzophenone glucopyranosides from Phaleria macrocarpa and related benzophenone glucopyranosides. AB - The first total syntheses of benzophenone glucopyranosides reported from Phaleria macrocarpa and related benzophenone glucopyranosides were successfully carried out. The alkoxy groups present ortho to the carbonyl group in polyalkoxybenzophenones were selectively deprotected by AlCl(3)-PhNMe(2) in high yields, leaving other alkoxy groups unaffected. It was concluded in the current synthetic study that all the reported benzophenone glucopyranosides possessed the same structure as 2,4',6-trihydroxy-4-methoxybenzophenone 2-O-beta-D glucopyranoside. PMID- 19809190 TI - Biosynthesis of indolocarbazole and goadsporin, two different heterocyclic antibiotics produced by actinomycetes. AB - The biosynthesis of staurosporine, rebeccamycin, and goadsporin, which are produced by actinomycetes and contain characteristic heterocyclic rings, was characterized by genetic methods. Staurosporine and rebeccamycin contain an indolocarbazole ring synthesized from two molecules of tryptophan, with indolepyruvic acid imine and chromopyrrolic acid as biosynthetic intermediates. A tetrameric hemoprotein synthesizes chromopyrrolic acid, and cytochrome P450 peroxidase catalyzes the intramolecular C-C coupling and decarboxylation of chromopyrrolic acid to form the indolocarbazole core. Goadsporin is a thiopeptide containing thiazole and oxazole heterocyclic rings. The structural gene godA is ribosomally translated to a goadsporin precursor peptide, and oxazole, methyloxazole, and thiazole rings are derived from serine, threonine, and cystein through post-translational modifications. On the basis of these knowledges, a wide variety of indolocarbazole and goadsporin analogs through the rational gene recombination and disruption of these biosynthetic genes were successfully produced. PMID- 19809191 TI - Accumulation of elicitor activity in the intercellular fluid of the Arabidopsis thaliana len3 mutant. AB - Activation of the hypersensitive response (HR) triggers localized acquired resistance (LAR) and systemic acquired resistance (SAR). Recently we found that Arabidopsis thaliana lesion initiation 3 (len3) plants develop lesions on leaves without pathogen attack, constitutively express PR genes, and accumulate elevated levels of SA. Hence we hypothesized that a signal important for LAR and SAR accumulates in the intercellular fluids (IFs) of the len3 plants. Infiltration of the IF from len3 plants induced PR-2 expression in local leaves but not in the systemic leaves of the wild type plants, suggesting that the elicitor activity of the IF contributes to LAR but not to SAR. Induction of PR-2 was dependent on SA signaling and ET signaling, and the elicitor in the IF was associated with molecules in the range of >100 kDa. These results suggest that len3 plants accumulate the elicitor in the IF, and that this might play a role in the establishment of LAR. PMID- 19809192 TI - The Corynebacterium glutamicum NCgl2281 gene encoding an RNase E/G family endoribonuclease can complement the Escherichia coli rng::cat mutation but not the rne-1 mutation. AB - The Corynebacterium glutamicum NCgl2281 gene encodes an RNase E/G family endoribonuclease having an additional N-terminal domain of unknown function. In this study, we constructed plasmids expressing the full length (FL) and the N terminally truncated form (DeltaN) of NCgl2281 and examined their complementation ability as to Escherichia coli rng::cat and rne-1 mutations. Both FL- and DeltaN NCgl2281 rescued the defects caused by the rng::cat mutation, i.e., accumulation of 16S rRNA precursor, overproduction of the AdhE protein, and growth inhibition on M9 glucose medium. On the other hand, they did not complement the rne-1 mutation. These results indicate that the C. glutamicum NCgl2281 endoribonuclease is functionally more closely related to the E. coli RNase G than to RNase E. PMID- 19809193 TI - Selective cytotoxicity of Ponciri Fructus against glucose-deprived PANC-1 human pancreatic cancer cells via blocking activation of GRP78. AB - Pancreatic cancer cells are sometimes exposed to stressful microenvironments such as glucose deprivation, hypoxia, and starvation of other nutrients. These stresses, which are characteristic of poorly vascularized solid tumors, activate the unfolded protein response (UPR). The UPR is a stress-signaling pathway present in tumor cells that is associated with molecular chaperone GRP78. Induction of GRP78 has been found to increase cell survival and decrease apoptotic potential through genetic alterations. Thus GRP78 may represent a novel target in the development of anticancer drugs. Here we established a novel screening program to identify chaperone modulators that exhibit preferential cytotoxic activity in glucose-deprived pancreatic cancer cells. During the course of our screening, we isolated an active substance, Ponciri Fructus (PF), from an herbal medicine source and identified it as a down-regulator of GRP78. As expected, PF inhibited expression of the GRP78 protein under glucose-deprivation conditions in a dose-dependent manner. Furthermore, it induced selective cytotoxicity against glucose-deprived cancer cells; this effect was not observed under normal growth conditions. We also detected apoptotic bodies on Hoechst staining and attempted to determine whether PF-induced apoptosis involved caspase 3 activation. Our results suggest that the GRP78-inhibitory action of PF was dependent on strict hypoglycemic conditions and that it resulted in the selective death of glucose-deprived pancreatic cancer cells. PMID- 19809194 TI - Changes in nitric oxide production levels and expression of nitric oxide synthase isoforms in the rat uterus during pregnancy. AB - We clarified nitric oxide (NO) production in the rat uterus by electron paramagnetic resonance spectroscopy and with Fe-N-(dithiocarboxy) sarcosine complex (an NO-trapping reagent). We examined changes in NO production in the whole uterus, decidua, and myometrium (gestational days 13.5-21.5). The expression of nitric oxide synthase (NOS) isoforms was also examined by quantitative reverse transcription-polymerase chain reaction. The uterine NO levels were low on day 13.5, peaked on day 17.5, and thereafter decreased significantly. The NO production levels in the decidua and myometrium were the same on day 13.5, but the levels in the decidua were 2- to 4-fold higher than those in the myometrium from day 15.5 onwards. The NOS-2 mRNA expression pattern correlated well with changes in the NO levels in the decidua, whereas the NOS-3 mRNA was expressed constantly during gestation. Thus NOS-2-generated NO in the decidua contributed significantly to uterine NO levels. PMID- 19809195 TI - Bone regeneration using an acellular extracellular matrix and bone marrow mesenchymal stem cells expressing Cbfa1. AB - To treat bone defects, tissue-engineering methods combine an appropriate scaffold with cells and osteogenic signals to stimulate bone repair. Mesenchymal stem cells (MSCs) derived from adult bone marrow are an ideal source of cells for tissue engineering, in particular for applications in skeletal and hard tissue repair. Core binding factor alpha1 (Cbfa1) is an essential transcription factor for osteoblast differentiation. However, the effects of Cbfa1 on MSCs in vitro and in vivo have not been well characterized. In this study, we found that MSCs modified genetically to express Cbfa1 promoted the healing of segmental defects of the radius in rabbits. First, osteogenic differentiation of MSCs transfected with an adenovirus encoding Cbfa1 was demonstrated. Expression of mRNA from a number of osteoblastic marker genes, including osteocalcin, osteopontin, and type I collagen, was detected. In addition, alkaline phosphatase activity and increased osteocalcin content were observed. The cells expressing the Cbfa1 gene were then combined with acellular bone extracellular matrix in a flow perfusion culture system. Finally, the cell-matrix constructs were implanted into radius defects in the rabbit model. After 12 weeks, radiographic, histological, and biomechanical analyses showed that MSCs modified with the Cbfa1 gene resulted in a significantly higher amount of newly-formed bone and rebuilding of the marrow cavity than control cell-matrix constructs. This study indicates that MSCs modified with the Cbfa1 gene can act as suitable seed cells for the regeneration of bone defects. PMID- 19809196 TI - Identification of red pigments formed in a D-xylose-glycine reaction system. AB - Blue, red, and yellow pigments were formed in the D-xylose (1 M)-glycine (0.1 M) reaction system. Novel red pigments were isolated and purified from the reaction solution, designated Red-M1 (red Maillard intermediate-1) and Red-M2 (red Maillard intermediate-2). Red-M1 was identified as 1,4,6,9-tetracarboxymethyl-5 (1,2,3,4-tetrahydroxybutyl)-8-hydroxymethyl-3-(2,3-dihydroxypropyl)-5,6-dihydro pyrrolo[2',3':4,5]pyrrolo[2,3-e]pyrrolo[3,2-b]azepine-9-ium. NMR and CD data indicated that Red-M2 was a diastereomer of Red-M1. They are assumed to be important Maillard reaction intermediates through the formation of melanoidins as well as blue pigments. PMID- 19809197 TI - Membrane topology and functional importance of the periplasmic region of ABC transporter LolCDE. AB - The LolCDE complex is an ATP-binding cassette transporter that mediates the release of newly synthesized lipoproteins from the cytoplasmic membrane of gram negative bacteria, which results in the initiation of outer-membrane sorting of lipoproteins through the Lol pathway. LolCDE is composed of one copy each of membrane subunits LolC and LolE, and two copies of nucleotide-binding subunit LolD. In this study, we examined the membrane topology of LolC and LolE by PhoA fusion analysis. Both LolC and LolE were found to have four transmembrane segments with a large periplasmic loop exposed to the periplasm. Despite similarities in sequence and topology, the accessibility of a sulfhydryl reagent to Cys introduced into the periplasmic loop suggested that the structure of the periplasmic region differs between LolC and LolE. Inhibition of the release of lipoproteins by the sulfhydryl reagent supported a previous proposal that LolC and LolE have distinct functions. PMID- 19809198 TI - Analysis of enzyme production by submerged culture of Aspergillus oryzae using whole barley. AB - We have reported on high enzyme production by submerged culture of Aspergillus kawachii using barley with the husk (whole barley). To elucidate the mechanism underlying this high enzyme production, we performed a detailed analysis. Aspergillus oryzae RIB40 was submerged-cultured using whole barley and milled whole barley. Enzyme production was analyzed in terms of changes in medium components and gene expression levels. When whole barley was used, high production of glucoamylase and alpha-amylase and high gene expression levels of these enzymes were observed. Low ammonium concentrations were maintained with nitrate ion uptake continuing into the late stage using whole barley. These findings suggest that the sustainability of nitrogen metabolism is related to high enzyme production, and that a mechanism other than that associated with the conventional amylase expression system is involved in this relationship. PMID- 19809199 TI - Ameyamaea chiangmaiensis gen. nov., sp. nov., an acetic acid bacterium in the alpha-Proteobacteria. AB - Two isolates, AC04(T) and AC05, were isolated from the flowers of red ginger collected in Chiang Mai, Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates were included within a lineage comprised of the genera Acidomonas, Gluconacetobacter, Asaia, Kozakia, Swaminathania, Neoasaia, Granulibacter, and Tanticharoenia, and they formed an independent cluster along with the type strain of Tanticharoenia sakaeratensis. The calculated pair-wise sequence similarities of isolate AC04(T) were 97.8-92.5% to the type strains of the type species of the 11 genera of acetic acid bacteria. The DNA base composition was 66.0-66.1 mol % G+C with a range of 0.1 mol %. A single-stranded, labeled DNA from isolate AC04(T) presented levels of DNA-DNA hybridization of 100, 85, 4, and 3% respectively to DNAs from isolates AC04(T) and AC05 and the type strains of Tanticharoenia sakaeratensis and Gluconacetobacter liquefaciens. The two isolates were unique morphologically in polar flagellation and physiologically in intense acetate oxidation to carbon dioxide and water and weak lactate oxidation. The intensity in acetate oxidation almost equaled that of the type strain of Acetobacter aceti. The two isolates had Q-10. Isolate AC04(T) was discriminated from the type strains of the type species of the 11 genera by 16S rRNA gene restriction analysis using restriction endonucleases TaqI and Hin6I. The unique phylogenetic, genetic, morphological, physiological, and biochemical characteristics obtained indicate that the two isolates can be classified into a separate genus, and Ameyamaea chiangmaiensis gen. nov., sp. nov. is proposed. The type strain is isolate AC04(T) (=BCC 15744(T), =NBRC 103196(T)), which has a DNA G+C content of 66.0 mol %. PMID- 19809200 TI - Molecular cloning and expression in Pichia pastoris of a Irpex lacteus exo-beta (1-->3)-galactanase gene. AB - A gene encoding exo-beta-(1-->3)-galactanase from Irpex lacteus was cloned by reverse transcriptase-PCR. The deduced amino acid sequence showed high similarity with exo-beta-(1-->3)-galactanases from other sources. The molecular mass of the mature form was calculated to be 45,520 Da. The gene product expressed in Pichia pastoris specifically hydrolyzed beta-(1-->3)-galactooligosaccharides, as did other exo-beta-(1-->3)-galactanases. The recombinant enzyme showed high activity toward arabinogalactan-proteins (AGPs) from radish as well as beta-(1-->3) galactan. Product analysis revealed that the enzyme released beta-(1-->6) galactobiose, beta-(1-->6)-galactotriose, and alpha-L-arabinofuranosyl-(1-->3) beta-galactosyl-(1-->6)-galactose together with Gal from beta-(1-->3)-galactans attached with and without beta-(1-->6)-galactosyl branches prepared from acacia gum. These results indicate that the exo-beta-(1-->3)-galactanase from I. lacteus efficiently hydrolyzes beta-(1-->3)-galactan main chains of AGPs by bypassing beta-(1-->6)-galactosyl side chains. PMID- 19809201 TI - Properties of fulvic acid extracted from excess sludge and its inhibiting effect on beta-hexosaminidase release. AB - The physicochemical and biological properties of fulvic acid extracted and purified from excess sludge and solubilized excess sludge were studied. Solubilization was introduced to improve the recovery rate of fulvic acid from the sludge. The structural features of fulvic acid from excess sludge and solubilized excess sludge were characterized by using an elemental analysis, Fourier transform infrared spectroscopy and (1)H-nuclear magnetic resonance spectroscopy, and were compared with fulvic acid extracted from peat which had an inhibitory effect on the type I allergy in our previous study. The results show that they had a higher aliphatic characteristic with lower oxygen group content than fulvic acid from peat, and that the aliphatic characteristic was further strengthened by the use of solubilization. The biological properties of fulvic acid from excess sludge and solubilized excess sludge showed an inhibitory effect on beta-hexosaminidase release at the antigen-antibody binding stage and antigen receptor binding stage by using rat basophilic leukemia cells. PMID- 19809202 TI - A functional analysis of the yeast ubiquitin ligase Rsp5: the involvement of the ubiquitin-conjugating enzyme Ubc4 and poly-ubiquitination in ethanol-induced down regulation of targeted proteins. AB - Rsp5 is an essential ubiquitin ligase in Saccharomyces cerevisiae. We have found that the Ala401Glu rsp5 mutant is hypersensitive to various stresses, suggesting that Rsp5 is a key enzyme for yeast cell growth under stress conditions. The ubiquitination and the subsequent degradation of stress-induced misfolded proteins are indispensable for cell survival under stress conditions. In this study, we analyzed the ubiquitin-conjugating enzyme Ubc4 and the poly ubiquitination of targeted proteins involved in the function of Rsp5 under ethanol stress conditions. Ubc4 was found to be important in yeast cell growth and poly-ubiquitination of the bulk proteins in the presence of ethanol. The general amino acid permease Gap1 is poly-ubiquitinated via Lys63 and is down regulated after the addition of ammonium ions through a process requiring Rsp5. We found that Gap1 was removed from the plasma membrane in the presence of ethanol in a Rsp5-dependent manner, and that the disappearance of Gap1 required Ubc4 and involved the lysine residues of ubiquitin. Our results also indicate that Lys6 of ubiquitin might inhibit the disappearance of Gap1. These results suggest that Rsp5 down-regulates the ethanol-induced misfolded forms of Gap1. In addition, it appears that the substrates of Rsp5 are appropriately poly ubiquitinated via different lysine residues of ubiquitin under various growth conditions. PMID- 19809203 TI - Acute myocardial infarction: the enduring challenge for cardiac protection and survival. AB - Although considerable advances have been made in the diagnosis and management of acute myocardial infarction (AMI), the disorder is still a major cause of morbidity and mortality worldwide and continues to pose significant therapeutic challenges. The use of biomarkers to aid the diagnosis of AMI is now increasing and has enabled better understanding of the pathophysiology of the disorder and identification of patients who require urgent reperfusion therapy. Early percutaneous coronary intervention (PCI) appears to be beneficial when performed in a timely manner with a door-to-balloon time <90 min. The goal of PCI is now shifting from simple revascularization of occluded coronary arteries to optimum reperfusion at the microvascular level. Effective strategies and pharmacological agents need to be developed for better cardiac protection during AMI. Most deaths resulting from AMI occur within 1 h of its onset, and half of them occur before hospital admission. Thus, an effective pre-hospital lifeline system should be an important priority, achieved through the chain of survival, including the immediate implementation of definitive resuscitative efforts and rapidly transporting the patients to the hospital. PMID- 19809204 TI - Efficacy of nebivolol on flow-mediated dilation in patients with slow coronary flow. AB - Slow coronary flow (SCF) is the phenomenon of slow progression of angiographic contrast in the coronary arteries in the absence of stenosis in the epicardial vessels in some patients presenting with chest pain. There are no definite treatment modalities for patients with SCF. Our aim was to investigate the efficacy of nebivolol in patients with slow coronary flow by monitoring its effects on endothelial function and different markers of inflammation. Forty-two patients (16 females, 26 males; mean age, 55 +/- 10) with slow coronary flow (SCF) were included in the study. After baseline assessment, the patients were administered nebivolol 5 mg once daily. After 12 weeks of nebivolol therapy, the biochemical and ultrasonographic examinations were repeated. Chest pain relief was detected in 38 patients after treatment (90%). Systolic and diastolic blood pressure and high sensitive CRP were significantly decreased after nebivolol therapy. Among brachial artery dilation variables that reflect endothelial function, basal resistive index (RI), post-flow mediated dilation RI, and post nitrate mediated dilation RI were significantly decreased after therapy. Nebivolol is effective at improving endothelial function in patients with SCF. It controls chest pain, decreases CRP, and has favorable effects on brachial artery dilation variables in patients with coronary slow flow. PMID- 19809205 TI - Additive antihypertensive and antihypertrophic effects of long-acting Ca blockers in uncontrolled hypertensive patients with angiotensin-receptor blocker based treatment. AB - The aim of the present study was to examine the antihypertensive and antihypertrophic effects of combined treatment with a long-acting calcium antagonist on top of an angiotensin II receptor blocker (ARB) in uncontrolled hypertensive patients. Patients with essential hypertension and a blood pressure > 140/90 mmHg on ARB monotherapy (losartan 50 mg/day or candesartan 8 mg/day) were randomly assigned to a nifedipine controlled release (CR) group (n = 15) or amlodipine group (n = 11). A significant additional antihypertensive effect was noted from 1 month with nifedipine and 2 months with amlodipine. The average daily dose was 25 mg for nifedipine and 5 mg for amlodipine. The cardiothoracic ratio was significantly reduced in both groups after 3 months. Left ventricular wall thickness and left ventricular mass index also decreased. Metabolic parameters, hepatic function, and renal function did not change significantly. Additional treatment with a long-acting calcium antagonist achieved further blood pressure reduction as well as an antihypertrophic effect in the uncontrolled patients with prior ARB monotherapy. PMID- 19809206 TI - Low HDL levels as the most common metabolic syndrome risk factor in heart failure. AB - Metabolic syndrome (MetS) includes some parameters which are associated with cardiovascular events and risk of developing heart failure (HF). The aim of the present study was to explore the prevalence of metabolic syndrome and individual MetS parameters among heart failure patients. Stable HF patients who had an ejection fraction (EF) < or = 35% were included. They were evaluated for MetS and parameters according to the Adult Treatment Panel III (ATP III) of the National Cholesterol Education Program (NCEP). One hundred and nine patients (72 males, 37 females; mean age, 67 +/- 12 years) were included. The most prevalent parameters were low HDL-C (69%) and hypertension (69%) in all participants. No significant change was observed in the prevalence of these parameters due to gender (P > 0.05). Hypertension, increased waist circumference, and hypertriglyceridemia were all significantly more common in women (P < 0.05). The prevalences of hypo-HDL emia, hyperglycemia, and hypertension did not differ significantly with an advanced age (P > 0.05), whereas hypertriglyceridemia and high waist circumference were significantly decreased in the elderly (P < 0 05). The overall MetS prevalence was 51% and it was significantly higher in women (76% versus 37%, P : 0.003). The prevalence of MetS clearly decreased with age, although the difference was not statistically significant (61% versus 46%, P : 0.57). The mean number of positive MetS parameters also changed significantly with age (3 +/- 1.4 versus 2.5 +/- 1.3, P : 0.046). EF did not change with mean number of MetS parameters (P > 0.05). Hypo-HDL-emia and similarly hypertension were the 2 most common MetS parameters in HF patients. Hypo-HDL-emia and hypertension were the most common parameters observed in all participants and no significant difference was seen due to gender or age. Even though the prevalence of MetS and the mean number of parameters were significantly more common in females and young patients, EF did not change with changes in these parameters. Based on the results obtained, we conclude that the early diagnosis and treatment of MetS as well as the measurement of individual parameters, especially the most frequent ones, may prevent heart failure or improve its status. PMID- 19809207 TI - The influences of red blood cell transfusion on perioperative inflammatory responses using a miniaturized biocompatible bypass with an asanguineous prime. AB - Whole blood transfusion generates an inflammatory response and may contribute to organ dysfunction following cardiopulmonary bypass. We established a miniaturized (minimum 140 mL) and biocompatible bypass system to reduce perioperative inflammatory responses and avoid blood transfusions. This study was designed to reveal the influences of stored red blood cell (RBC) transfusions on perioperative inflammatory responses in infants. Fifty-four consecutive patients weighing 4-10 kg who underwent surgical procedures for complex heart anomalies with asanguineous prime were retrospectively reviewed. Twenty-two patients (40.7%) received RBC transfusions during CPB. The postoperative peak white blood cell count (p-WBC), peak neutrophil count (p-NC), and peak C-reactive protein (p CRP) were compared for both patient groups. Stepwise multiple logistic regression analyses were used to investigate which of the factors most affected the perioperative inflammatory responses. The p-CRP and p-NC in patients with transfusion was significantly greater than those in patients without transfusion (CRP 8.1 +/- 5.1 versus 5.5 +/- 3.0 mg/dL, P < 0.05, p-NC 14.6 +/- 4.5 versus 12.0 +/- 4.0 x 1000/mm3, P < 0.05). Multivariate analyses showed that RBC transfusion most affected p-WBC (coefficient: 3.89, 95% confidence interval [CI] 0.79-6.99, P = 0.015) and p-NC (coefficient: 3.64, 95% CI 0.87-6.40, P = 0.011). The RBC transfusions increased the perioperative inflammatory responses, compared to transfusion-free procedures, even when using a miniaturized biocompatible bypass with an asanguineous prime. PMID- 19809208 TI - Procoagulant state in heart failure with preserved left ventricular ejection fraction. AB - The impact of heart failure with preserved left ventricular ejection fraction (LVEF) on activated hemostasis is still unclear. We sought to compare the activation of hemostasis in patients with heart failure with preserved LVEF, with impaired LVEF, and in healthy controls. Biomarkers of coagulation and fibrinolysis (D-dimer, tPA and PAI-1) were determined in outpatients with chronic stable (NYHA I-III), optimally managed heart failure with preserved LVEF (n = 46) and with impaired LVEF (n = 52), and in healthy age- and gender-matched controls (n = 14). In comparison to healthy controls, patients with heart failure and preserved LVEF had increased median D-dimer levels (606 [330-1222] microg/L versus 174 [86-249] microg/L; P < 0.001), and median PAI-1 (20 [15.3-33.1] microg versus 6.2[3.4-8.9] microg/L; P < 0.001) and tPA antigen concentrations (9.6 [8.1 13.3] versus 3.6 [2.2-5.0] microg/L; P < 0.001). However, unlike tPA and PAI antigens, D-dimer levels in preserved LVEF did not reach values as high as in impaired LVEF (917 [454-1185] microg/L; P = 0.013). Moreover, in patients with impaired LVEF, but not in those with preserved LVEF, age and NT-proBNP emerged as independent predictors of log-transformed D-dimer levels. Heart failure with preserved LVEF is associated with a procoagulant state as determined by increased levels of D-dimer, tPA and PAI-1 antigens. D-dimer levels are significantly higher in patients with impaired LVEF, while tPA and PAI-1 levels are increased regardless of LVEF. PMID- 19809209 TI - Is allogeneic blood transfusion a risk factor for sternal dehiscence following cardiac surgery? A prospective observational study. AB - Sternal dehiscence following cardiac surgery has a multifactorial etiology. Significant risk factors contributing to sternal dehiscence include chronic obstructive pulmonary disease (COPD), obesity, or re-exploration due to bleeding or pericardial tamponade. We have focused on the role of allogeneic blood transfusion as a factor leading to the poor healing of surgical wounds. A prospective observational study of 1553 elective and emergency cardiac surgery patients was performed between January 2003 and June 2007. All of the patients enrolled in this study underwent median sternotomy. We studied the relationship between sternal dehiscence following cardiac surgery and the total number of packed red blood cells transfused. The incidence of sternal dehiscence in the study group was 3.4%. Diabetic patients did not have a higher incidence of wound dehiscence. Although COPD, obesity, and re-exploration contributed to sternal dehiscence, the number of allogeneic blood transfusions during the perioperative period was an important independent risk factor for sternal dehiscence. Patients with sternal dehiscence received an average of 7.6 transfusion units (TU) of allogeneic blood versus 1.6 TU of allogeneic blood in the group without sternal dehiscence (P < 0.00005). The dehiscence affected patients without any other significant risk factor who received 6 or more TU, or patients with at least one significant risk factor who received 4 or more TU of allogeneic blood. According to our results, the total amount of allogeneic blood transfused is an important risk factor contributing to sternal dehiscence. Regardless of other risk preconditions, the transfusion of 6 or more TU could result in sternal dehiscence following cardiac surgery. PMID- 19809210 TI - Prevalence and prognosis of patients with heart failure in Tokyo: a prospective cohort of Shinken Database 2004-5. AB - Prognosis of patients with heart failure (HF) remains unclear in Japan and should be determined in a prospective fashion. A prospective cohort of The Shinken Database comprised details on all of the new patients, including both inpatients and outpatients, who visited The Cardiovascular Institute Hospital in 2004-2005. HF patients were defined as those with symptomatic HF coexisting with structural heart diseases. Among 4,255 patients who visited our hospital, 597 patients (male/female 414/183, age 65.1 +/- 12.9 years, LVEF 56.2 +/- 18.0%) were diagnosed as presenting symptomatic HF. Ischemic heart disease was present in 305 (51.1%), valvular heart disease in 212 (35.5%), dilated cardiomyopathy in 59 (9.9%), hypertrophic cardiomyopathy in 24 (4.0%), hypertensive heart disease in 14 (2.3%), and others in 67 (11.2%). Hypertension, atrial fibrillation, and diabetes were observed in 35.3%, 27.4%, and 23.7%, respectively. During the mean follow-up period of 539 +/- 257 days, 40 deaths (5.0% per year) occurred, including 34 cardiovascular deaths (4.5% per year, NYHA class II: 1.0%, III: 11.3%, IV: 36.6% per year, respectively). The present study showed that the prognosis of Japanese patients with HF among moderate to severe severity was found to be similar to that of Western countries. Multiple Cox hazard analysis identified the presence of chronic kidney disease and NYHA class as independent predictors for cardiovascular death. This prospective cohort study identified the prevalence, prognosis, and risk factors in HF patients to provide a basis for therapeutic management in Japan. PMID- 19809211 TI - Influences of autonomic nervous system on atrial arrhythmogenic substrates and the incidence of atrial fibrillation in diabetic heart. AB - Diabetes mellitus (DM) is clinically associated with an increased incidence of atrial fibrillation (AF), but the underlying mechanism remains unclear. We hypothesized that neural remodeling enhances AF vulnerability in diabetic hearts. Eight weeks after creating streptozotocin-induced diabetic rats (DM rats) or control rats, the hearts were perfused according to the Langendorff method. Inducibility of AF was evaluated by 5 times burst pacing from the right atrium and the atrial effective refractory period (AERP) was measured. The protocol was repeated during sympathetic nerve stimulation (SNS) or parasympathetic nerve stimulation (PNS). In tissue samples taken from the right atrium, the density of nerves positive for tyrosine hydroxylase (TH) and acetylcholinesterase (AChE) were determined. SNS significantly increased the incidence of AF in DM rats (14 +/- 6 to 30 +/- 8%, P < 0.01), but not in control rats (11 +/- 4 to 14 +/- 6%, NS). Although AERP was significantly decreased by SNS in both rats (each P < 0.01), increased heterogeneity of AERP by SNS was seen only in DM rats. PNS significantly decreased AERP and increased the incidence of AF (9 +/- 5 to 30 +/- 5% in control rats, 12 +/- 6 to 27 +/- 6% in DM rats, each P < 0.01) in both rats. The density of TH-positive nerves was heterogeneous in DM rats compared with control rats, whereas the heterogeneity of AChE-positive nerves was not different in the rats. The prevalence of AF was enhanced by adrenergic activation in diabetic hearts, in which heterogeneous sympathetic innervation was evident. These results suggest that neural remodeling may play a crucial role for increased AF vulnerability in DM. PMID- 19809212 TI - Developed force of papillary muscle: what index correctly indicates contractile capacity? AB - We hypothesized that similar samples of the same normal heart should report similar contractile index values. We analyzed anterior (AP) and posterior (PP) papillary muscles (PM) of the same heart (n = 46), whose representation of force fulfills this premise calculating force (F: mN), tension (T: mN/mm2), and tension per milligram of myocardium (delta: mN/mm2/mg). In all analyses, F and +dF/dt as well as T and dT/dt values were higher in heavier PM. These differences disappeared for delta and ddelta/dt. There was a significant and positive correlation for F and T as well as its derivative with myocardial mass. Myocardial depression (verapamil) of PP, in comparison to AP, was not recognized by F or T, but was identified when reported as delta. We conclude that the normalization of tension for papillary muscle mass is the most appropriate form for reporting intrinsic contractile capacity in PM since F and T depend on the myocardial mass participating in contraction. PMID- 19809213 TI - Culturing of ventricle cells at high density and construction of engineered cardiac cell sheets without scaffold. AB - In natural heart tissue, cell density is about 1.0 x 108/cm3, and the cell metabolism is very active. Therefore, culturing heart cells in 3-dimensions at high density and construction of engineered cardiac tissue in vitro is very difficult. The aim of this study was to simulate 3-dimensional culturing of cardiac cells and pursue a novel method to construct engineered cardiac tissue in vitro. The isolated neonatal rat ventricle cells were cultured at a high seeding density of 1 x 10(6)/cm2. The cells at high density metabolized actively; the glucose consumption and lactic acid production of ventricle cells were much greater than those of fibroblasts cultured at the same density. The pH value of the culture medium of ventricle cells consistently decreased more rapidly. These cultured ventricle cells contained vascular endothelial cells, cardiomyocytes, and smooth muscle cells that appeared close to each other, and had overlapping nuclei and plenty of extracellular collagen. The cells at high density were treated with 0.2% trypsin to construct engineered cardiac cell sheets without scaffold. The engineered cardiac cell sheets could beat and roll up spontaneously, each sheet was 3 to 5 cells thick, and contained abundant cardiomyocytes and extracellular collagen. In conclusion, cells cultured at high density in vitro grew well in a 2-dimensional culturing environment, formed "quasi 3-dimension" culturing, and engineered cardiac cell sheets comprised of several layers of cells were constructed. This study provides some guidance for cardiac tissue engineering and a novel method to construct engineered cardiac tissue without scaffold. PMID- 19809214 TI - Simultaneous right and left coronary occlusion caused by an extensive dissection to the coronary sinus of Valsalva during percutaneous intervention in right coronary artery. AB - We report a case of simultaneous right and left coronary occlusion during percutaneous coronary intervention in the right coronary artery. An aortocoronary dissection induced by the forceful manipulation extended from the right to left sinus of Valsalva and occluded the ostia of both coronary arteries. The patient suffered cardiogenic shock and ventricular fibrillation. However, after successful rapid stenting to right and left coronary arteries, safe discharge was possible. PMID- 19809215 TI - A case of Brugada syndrome in which diurnal ECG changes were associated with circadian rhythms of sex hormones. AB - We describe a thought-provoking case of Brugada syndrome in which a relationship between the diurnal electrocardiogram (ECG) changes and sex hormone levels was observed. A 36-year-old man who experienced cardiac arrest was referred to our hospital. He had a family history of sudden cardiac death. The 12-lead ECG exhibited a mild coved type ST-segment elevation in leads V1 and V2, which was enhanced by intravenous pilsicainide injection. Hence, this case was diagnosed as Brugada syndrome. The circadian rhythm of the serum testosterone level revealed low levels in the daytime (1.66-1.99 ng/mL) and high levels (2.52-3.42 ng/mL) in the nighttime. Interestingly, augmentation of the ST segment elevation and widening of the P wave were observed at around 2:00 AM, when the serum testosterone was recorded at its highest. Our report discusses the influence of the circadian rhythms of sex hormones on the ECG changes in Brugada syndrome. PMID- 19809216 TI - Left bundle branch block with intermittent QRS axis switching: observation of a hypertensive patient for 18 years. AB - A 64-year-old man who had been prescribed antihypertensive drugs since 1971 attended our clinic in 1988 with hypertension and electrocardiographic abnormalities. An electrocardiogram revealed left axis deviation (LAD) in 1988 and slightly prolonged PQ intervals in 1993. Complete left bundle branch block (CLBBB) with LAD developed in May 1995. The wide QRS of the CLBBB had never returned to the normal narrow QRS and had intermittently alternated between LAD and normal axis. The PQ intervals were longer when the QRS axis showed LAD compared to that with normal QRS axis. The QRS complexes in leads V1-V3 revealed an R wave at LAD and a QS pattern at normal axis. During a deep breathing test, the QRS axis switched from normal axis to LAD at the end of forced expiration and also switched from normal axis to LAD within a few minutes after the exercise test. These results suggest that the shift of the QRS axis might be related to the tone of the autonomic nervous system. PMID- 19809217 TI - A case of early phase dialysis associated effusive constrictive pericarditis with distinct surgical findings. AB - A 55-year-old male patient with a 10 year history of hemodialysis was admitted for examination of pericardial effusion. Specific echocardiography, MRI, and cardiac catheterization findings strongly suggested a diagnosis of effusive constrictive pericarditis. Pericardiectomy showed the following distinct findings. Two layers of fibrous tissue below the thickened pericardium emerged. The cardiac constriction had not been relieved until the inner layer was partially resected. We conclude that patients in dialysis associated constrictive pericarditis should undergo pericardiectomy before the adhesion between fibrous tissues becomes hard. PMID- 19809218 TI - Peroxisome proliferators attenuate free arachidonic acid pool in the kidney through inducing lysophospholipid acyltransferases. AB - Attenuating effects of peroxisome proliferators on the concentration of free arachidonic acid by inducing 1-acyl-2-lysophospholipid acyltransferases in the kidney were studied. The administration of the three structurally dissimilar peroxisome proliferators, 2-(4-chlorophenoxy)-2-methylpropionic acid (clofibric acid), di(2-ethylhexyl)phthalate, and 2,2'-(decamethylenedithio)diethanol, to rats or mice considerably increased the activities of microsomal 1 acylglycerophosphoethanolamine acyltransferase (LPEAT), 1 acylglycerophosphoinositol acyltransferase (LPIAT), 1-acylglycerophosphoserine acyltransferase (LPSAT), and 1-acylglycerophosphocholine acyltransferase (LPCAT), and the mRNA level of LPCAT3, but not the mRNA level of LPCAT1, LPCAT4, or LPEAT1, in the kidney and the liver. The proportions of arachidonic acid in phospholipids in renal microsomes are rather high for the low proportion of arachidonic acid in free fatty acids in renal microsomes of control rats. The treatment of rats with clofibric acid attenuated the concentration and the proportion of free arachidonic acid to about a half; nevertheless the treatment lowered slightly the proportions of arachidonic acid in phospholipids other than phosphatidylcholine. These results indicate that peroxisome proliferators upregulate the four 1-acyl-2-lysophospholipid acyltransferases of the kidney and, and the induced 1-acyl-2-lysophospholipid acyltransferases seem to play a physiologically crucial contribution in attenuating the pool of free arachidonic acid in the kidney. PMID- 19809219 TI - Effects of telmisartan on right ventricular remodeling induced by monocrotaline in rats. AB - The present study investigated whether telmisartan, an angiotensin II type 1 receptor antagonist, has cardioprotective effects on monocrotaline-induced right ventricular (RV) remodeling in rats. Six-week-old male Wistar rats were divided into control group (CONT), monocrotaline (60 mg/kg, i.p.)-treated group (MCT), monocrotaline (60 mg/kg, i.p.) + telmisartan (3 mg/kg per day, p.o.)-treated group (MCT+TEL), and telmisartan (3 mg/kg per day, p.o.) alone-treated group (TEL). Hearts were excised after echocardiography examinations at day 25. Significant increase in RV weight and histologically remarkable fibrosis in RV sections were observed in MCT. Tricuspid annular plane systolic excursion, a parameter for RV systolic function, significantly decreased in MCT. These RV hypertrophy, fibrosis, and dysfunction were inhibited in MCT+TEL. In MCT, the acceleration time/ejection time ratio of pulmonary artery flow velocity, an index of pulmonary hypertension, significantly decreased. This decrease was not affected in MCT+TEL. In MCT, expressions and activities of matrix metalloproteinase (MMP)-2 and MMP-9, which play a critical role in cardiac remodeling, significantly increased in the RV. In MCT+TEL, these increases in expressions and activities were inhibited. MCT showed about 2-fold increase in transforming growth factor-beta1 expression compared with CONT, and such an increase was not decreased in MCT+TEL. There were no significant changes of these parameters in TEL compared with CONT. These results suggest that telmisartan could attenuate the monocrotaline-induced RV remodeling through improvements of RV hypertrophy, fibrosis, dysfunction, and inhibition of MMPs. PMID- 19809220 TI - Potent antidiabetic effects of rivoglitazone, a novel peroxisome proliferator activated receptor-gamma agonist, in obese diabetic rodent models. AB - The pharmacological effects of rivoglitazone, a novel thiazolidinedione derivative peroxisome proliferator-activated receptor (PPAR)-gamma agonist, were characterized in vitro and in vivo. Rivoglitazone activated human PPARgamma more potently compared with rosiglitazone and pioglitazone and had little effect on PPARalpha and PPARdelta activity in luciferase reporter assays. In Zucker diabetic fatty (ZDF) rats, 14-day administration of rivoglitazone decreased the plasma glucose and triglyceride (TG) levels in a dose-dependent manner. The glucose-lowering effect of rivoglitazone was much more potent than those of pioglitazone (ED(50): 0.19 vs. 34 mg/kg) and rosiglitazone (ED(50): 0.20 vs. 28 mg/kg). In addition, rivoglitazone showed potent antidiabetic effects in diabetic db/db mice. In Zucker fatty rats, rivoglitazone at a dose of 0.1 mg/kg clearly ameliorated insulin resistance and lowered plasma TG levels by accelerating the clearance of plasma TG. Gene expression analysis in the liver and heart of ZDF rats treated with rivoglitazone for 14 days suggested that rivoglitazone may reduce hepatic glucose production and modulate the balance of the cardiac glucose/fatty acid metabolism in diabetic animals. In summary, we showed that rivoglitazone is a potent and selective PPARgamma agonist and has a potent glucose-lowering effect via improvement of the insulin resistance in diabetic animal models. PMID- 19809221 TI - Immunodetection of NGF, trkA, p75 and inhibin alpha-subunit in interstitial cells of golden hamsters treated with hCG. AB - The aim of this study was to investigate the effect of luteinizing hormone (LH) surge on expression of nerve growth factor (NGF), trkA, p75 and inhibin alpha subunit in ovarian interstitial cells of golden hamsters after human chorionic gonadotropin (hCG) treatment. NGF, two NGF receptors (trkA, p75) and inhibin alpha-subunit were immunolocalized by immunohistochemistry, and gonadotropins, steroid hormones and immunoreactive (ir-) inhibin concentrations were measured by radioimmunoassay. Stronger positive staining of NGF, trkA and p75 were found in interstitial cells at 6, 12 and 18 h after hCG injection in the treated group compared with the control groups. Inhibin alpha-subunit staining was found in interstitial cells at 12 and 18 h after hCG injection in the treatment group, but not in the control group. Plasma concentrations of progesterone increased significantly from 6 to 18 h after hCG treatment, whereas plasma concentrations of estradiol-17beta significantly decreased compared with the control group. An increased plasma concentration of FSH and decreased concentrations of ir-inhibin from 6 to 18 h after hCG treatment verified the negative relationship. There were no significant changes in the concentrations of LH in the hCG-treated group. In contrast, the typical preovulatory LH surge was found at 1700 h on day 4 (proestrus) in the control group. These results suggested that LH surge can induce expression of NGF, trkA, p75 and inhibin alpha-subunit in ovarian interstitial cells and that NGF, trkA, p75 and inhibin alpha-subunit may have a paracrine or autocrine role in modulation of ovarian interstitial cell function in golden hamsters. PMID- 19809222 TI - Effect of N-acetyl-D-glucosamine on bovine sperm-oocyte interactions. AB - N-Acetyl-D-glucosamine (GlcNAc) is a major component of glycosaminoglycan, which is involved in sperm-oocyte interactions. We examined the effect of adding GlcNAc and other monosaccharides, D-mannose and D-fucose, to the in vitro fertilization (IVF) medium on bovine sperm-oocyte interactions. In medium in which sperm and a zona pellucida (ZP) were co-incubated with monosaccharides for 5 min, addition of GlcNAc (5 or 25 mM) significantly reduced the number of sperm that attached to the ZP. Pretreatment of gametes with GlcNAc (5 mM) prior to co-incubation also suppressed sperm-ZP attachment. Addition of GlcNAc (5 or 25 mM) to the medium in which sperm and a ZP were co-incubated for 5 h, however, significantly increased the number of sperm binding to and penetrating the ZP in a concentration-related manner. The other monosaccharides, D-fucose and D-mannose, did not have this effect. Supplementation of the sperm-oocyte co-incubation medium with 5 mM GlcNAc also enhanced the rate of polyspermic fertilization. When the ZPs were removed from the oocytes, GlcNAc did not affect the fertilization rate. Furthermore, incubation of sperm with 5 mM GlcNAc induced sperm membrane destabilization and an acrosome reaction, as evidenced by the hypo-osmotic swelling test and fluorescein isothiocyanate-labeled peanut agglutinin/propidium iodide (FITC PNA/PI) staining. Finally, GlcNAc suppressed ZP hardening following fertilization, as determined by measuring the time required for pronase to dissolve the ZP. In conclusion, supplementation of IVF medium with GlcNAc has various effects on sperm-oocyte interactions including suppression of initial attachment, induction of sperm membrane destabilization and acrosome reaction, increase in the number of sperm secondarily bound to and penetrating the ZP, suppression of ZP hardening following sperm-oocyte co-incubation and increase in the rate of polyspermic fertilization. PMID- 19809223 TI - Development of a noninvasive monitoring system for evaluation of Oct-3/4 promoter status in miniature pig somatic cell nuclear transfer embryos. AB - The present study was carried out to develop a noninvasive monitoring system for evaluation of Oct-3/4 promoter gene status in miniature pig somatic cell nuclear transfer (SCNT) embryos during in vitro development. Miniature pig fetal fibroblasts (MPFFs) were transfected with a gene construct consisting of two expression units, a mouse Oct-3/4 promoter-driven enhanced green fluorescent protein (EGFP) gene (EGFP expression only detected in Oct-3/4-expressing cells) and a neomycin resistance gene. After neomycin selection, MPFFs that did not express EGFP were fused with enucleated pig oocytes, cultured in vitro and assessed for EGFP expression. EGFP expression was detectable in all morulae (at 4 6 days of culture) and 50.0% of blastocysts (at 5-6 days of culture), whereas none of the 1-cell to 16-cell embryos at 1-5 days of culture expressed EGFP. On the other hand, EGFP expression was not maintained in all blastocysts at 7 days of culture. The reactivity with anti-Oct-3/4 antibodies also peaked from the morula to blastocyst stages at 5 days of culture. The results showed that reactivation of the Oct-3/4 promoter gene of donor nuclei occurs in the morula to blastocyst stages at 4-6 days after SCNT and that this noninvasive monitoring system using Oct-3/4 promoter-driven EGFP gene would be useful for evaluation of the reprogramming status of donor nuclei. PMID- 19809224 TI - [All-inside meniscal repair using the RapidLoc device]. AB - OBJECTIVES: Although conventional suture techniques yield satisfactory results in the treatment of meniscal ruptures, they are inherent with long operative time and high complication rates. The purpose of this study was to evaluate the results of meniscal repair with the use of the RapidLoc device. METHODS: The study included 57 consecutive patients (all males; mean age 24 years; range 17 to 33 years) who underwent meniscal repair with the RapidLoc device. The mean time from injury to surgery was 20 days (range 7 to 60 days). The mean length of meniscal ruptures was 25 mm (range 10 to 35 mm). A mean of two RapidLoc fixators were used for each rupture. Thirty patients had associated anterior cruciate ligament (ACL) rupture; of whom 17 patients underwent ACL reconstruction with hamstring autografts. Thirteen patients refused ACL reconstruction and underwent only meniscal repair. Functional results were evaluated using the Tegner activity score, Lysholm score, and IKDC (International Knee Documentation Committee) subjective knee evaluation form. Clinical assessments were made using the Barrett criteria. The mean follow-up was 39 months (range 18 to 66 months). RESULTS: The mean operation time including diagnostic and surgical arthroscopy was 25 minutes (range 15 to 35 min). Compared to the preoperative scores, all functional scores showed significant improvements (p<0.001) with the following increases: Lysholm score from 58.8 + or - 13.7 to 93.3 + or - 7.7, Tegner activity score from 3.5 + or - 1.0 to 6.0 + or - 1.6, and IKDC score from 52.8 + or - 10.5 to 91.4 + or - 4.0. According to the Barrett criteria, meniscal healing was achieved in all the patients. Second-look arthroscopy was performed in one patient due to pain and swelling, which showed an unabsorbed tophat of the RapidLoc device. CONCLUSION: Our findings suggest that the RapidLoc meniscal repair device may be an appropriate choice in selected cases because of satisfactory short-term clinical results, ease of applicability, short operation time, and low complication rate. PMID- 19809225 TI - [Comparison of phenol applications of different durations for the cauterization of the germinal matrix: an efficacy and safety study]. AB - OBJECTIVES: Partial nail avulsion with phenol matricectomy is one of the most widely performed procedures for the treatment of ingrowing nails. We compared phenol applications of different durations with respect to efficacy and safety. METHODS: The study included 148 ingrowing nails (grade 2-3) of 110 patients (54 males, 56 females). The patients were randomized to three groups for 1-, 2-, and 3-minute applications of phenol cauterization of the germinal matrix following surgical removal of ingrowing nails. Postoperative evaluations were made on days 2, 10, 16, 24, and 30 for pain, drainage, and tissue damage. Recurrences were recorded during a follow-up of 24 months. RESULTS: Improvements in pain, drainage, and tissue damage in each group were significant (p<0.001). Time to complete healing and durations of drainage and tissue damage were significantly shorter in patients receiving 1-minute phenol application (p<0.001), and the remaining groups did not differ in this respect. The mean duration of pain was similar in three groups. While the frequencies of pain and tissue damage were similar in three phenol groups, the number of patients having drainage on day 16 was significantly lower with 1-minute phenol application (p<0.001). All recurrences appeared in the sixth month and there was no significant difference between the three groups with respect to the recurrence rate. CONCLUSION: Our findings suggest that 1-minute phenol cauterization of the germinal matrix has a better safety profile than prolonged applications in the treatment of ingrown nails. PMID- 19809226 TI - [Volar locking plate fixation of unstable distal radius fractures]. AB - OBJECTIVES: We evaluated radiographic and functional results of volar locking plate fixation of unstable distal radius fractures. METHODS: The study included 27 patients (15 males, 12 females; mean age 45 years; range 18 to 77 years) who were treated with volar locking plate fixation for unstable distal radius fractures. Twenty-two patients (81.5%) had AO type C and five (18.5%) had type B fractures. Two patients (7.4%) had type I open fractures. Dorsal and volar angulation were present in 21 (77.8%) and six (22.2%) fractures, respectively. Four patients (14.8%) also had elbow dislocation and/or fracture and nine patients (33.3%) had disruption of the distal radioulnar joint. Autologous iliac crest graft was used in 10 patients (37%). Postoperative assessments included range of movement and grip strength measurements, and radiographic parameters using the Stewart criteria. Functional results were assessed using the Q-DASH (Quick-Disabilities of the Arm, Shoulder and Hand) questionnaire and the Gartland Werley scale. The mean follow-up was 18 months (range 12 to 34 months). RESULTS: All fractures united without a problem within a mean of 6.5 weeks (range 6 to 8 weeks). The following parameters became equal to the unaffected side: ulnar variance (n=21, 77.8%), radial inclination angle (n=13, 48.2%), radial tilt angle (n=8, 29.6%), and radial height (n=11, 40.7%). The mean radiographic Stewart score was 0.5 (range 0 to 3). The mean grip strength of the operated side was 72.4% (18 kg) of the normal side. The mean Q-DASH and Gartland-Werley scores were 8.3 (range 0 to 70.5) and 4.7 (range 0 to 16), respectively. According to the Gartland-Werley scale, the results were excellent in 12 patients (44.4%), good in 12 patients, and moderate in three patients (11.1%). Three complications (11.1%) occurred, including loss of alignment in two patients, and screw-related discomfort in one patient. CONCLUSION: Volar locking plate fixation is an effective treatment in the anatomical and functional restoration of unstable distal radius fractures. PMID- 19809227 TI - [The results of treatment for isolated zone 3 extensor tendon injuries]. AB - OBJECTIVES: We evaluated late-term results of surgical repair and physical rehabilitation of isolated zone 3 extensor tendon injuries. METHODS: Sixteen patients (13 males, 3 females; mean age 28 years; range 11 to 57 years) underwent surgical repair for isolated simple central slip injuries of zone 3 extensor tendon. Injuries involved the second, third, fourth, and fifth fingers in seven, three, two, and four patients, respectively. Primary tendon repair was performed in 14 patients. Two patients were treated after one week and two months following primary injury, respectively. Tendons were repaired by modified Kessler and epitendinous sutures in four patients, and by locking running suture in 12 patients. Following surgical repair, three patients underwent K-wire fixation in extension for two weeks, while 13 patients received the short arc motion protocol. The patients were assessed with respect to proximal interphalangeal (PIP) joint motion and extension loss. Functional results were assessed using the Strickland formula. The mean follow-up was 58 months (range 8 to 120 months). RESULTS: Full range of motion of the PIP joint was achieved in 15 patients (93.8%). The mean PIP joint motion was 98 degrees. One patient (6.3%) had an extension loss of 10 degrees in the range of motion of the PIP joint. There were no losses in the range of motion in three patients treated with K-wire fixation. The mean of the Strickland formula was 94.8% (range 74% to 100%), showing an excellent result in 15 patients (98.3%) and a good result in one patient (6.3%). All the patients returned to their pre-injury work status. None had buttonhole deformity or soft tissue complication. CONCLUSION: A proper suture technique combined with the short arc motion protocol provides good results in the treatment of isolated zone 3 extensor tendon injuries. PMID- 19809228 TI - [Open reduction and low-profile plate and/or screw fixation in the treatment of phalangeal fractures]. AB - OBJECTIVES: We evaluated the functional results and the effectiveness of open reduction and low-profile plate and/or screw fixation in the treatment of phalangeal fractures. METHODS: The study included 17 patients (5 women, 12 men; mean age 33 + or - 10 years; range 17 to 48 years) with closed, nonarticular phalangeal fractures. One patient had two phalangeal fractures. There were 14 proximal and four mid phalangeal fractures, including seven oblique, four spiral, one transverse, and six comminuted fractures. The mean duration from injury to surgery was 2.6 + or - 2.9 days (range 0 to 11 days). Following open reduction, seven fractures were treated with a mini plate and screws, and 11 fractures with mini screws. For functional evaluations, total active motion (TARM) and grip strength were measured and the Q-DASH (Quick-Disabilities of the Arm, Shoulder and Hand) questionnaire was administered. The mean follow-up was 35 + or - 20 months (range 12 to 75 months). RESULTS: Union was obtained in all the patients in a mean of 4.5 months. At final assessments, TARM ranged from 160 degrees to 260 degrees (mean 200 + or - 39.5 degrees ), showing excellent, good, and moderate results in six fingers (33.3%), five fingers (27.8%), and seven fingers (38.9%), respectively. An excellent or good TARM was obtained in eight fingers (72.2%) treated with mini screws, and in three fingers (42.9%) treated with plate screw fixation. The mean grip strength decreased by 7.5% (range 0 to 20%) on the affected side. The mean Q-DASH score was 3.4 + or - 4.4. According to the classification of complications proposed by Page and Stern, seven major (38.9%) and six minor (33.3%) complications were seen. Two patients (11.1%) had delayed union. CONCLUSION: Our findings do not justify open reduction and low-profile plate and/or screw fixation as the first choice in the treatment of phalangeal fractures. If plate and screw fixation is necessary, the most minimally invasive method such as screw fixation should be preferred. PMID- 19809229 TI - [An alternative fixation method for the treatment of unstable distal clavicle fractures: locked distal radius plate]. AB - OBJECTIVES: Optimal surgical fixation method for displaced distal clavicle fractures should not impose limitations on neighboring joint movements. We evaluated the results of surgical treatment of displaced distal clavicle fractures using locked distal radius plates. METHODS: Displaced distal clavicle fractures of 14 consecutive patients (11 men, 3 women; mean age 30 + or - 9 years; range 19 to 51 years) were treated using open reduction and locked distal radius plates. Before final fixation, one patient underwent K-wire fixation with tension band at another center, resulting in nonunion. Except for two cases with late presentation, the mean time to surgery was 5.3 days (range 1 to 17 days). According to the Neer classification, fresh fractures were type II in 10 patients and type III in three patients. Shoulder examinations and functional evaluations were made at 3, 6, and 12 months postoperatively. Functional assessment included the Modified Shoulder Rating Scale and Constant score. RESULTS: All patients achieved full range of motion of the shoulder at six weeks postoperatively. The mean modified shoulder score was 18.7 + or - 1.5 and the mean Constant score was 95.4 + or - 3.0 at 12 months. None of the patients developed implant failure, loss of reduction, skin breakdown, or infection. CONCLUSION: In selected acute fractures and nonunions of the distal clavicle, excellent clinical results are easily achievable with locked distal radius plate fixation because it allows early shoulder movements without necessitating implant removal. PMID- 19809230 TI - [The contribution of subacromial injection to the conservative treatment of impingement syndrome]. AB - OBJECTIVES: We evaluated the contribution of subacromial local anesthetic and corticosteroid injection to the conservative treatment of subacromial impingement syndrome. METHODS: The study included 56 patients (39 women, 17 men; mean age 50 years; range 31 to 68 years) with subacromial impingement syndrome without any rotator cuff lesion. The patients were randomly allocated to injection and control groups equal in number. The former group received a single subacromial injection of 9 ml bupivacaine and 1 ml betamethasone at the beginning of the treatment. The same physical therapy and rehabilitation program was administered to both groups, consisting of 15 sessions (3 weeks). Evaluations were made before, and three and six weeks after treatment. Functional results were assessed using the Constant score, pain was assessed using a visual analog scale, and range of motion was measured with a goniometer. RESULTS: Compared to pretreatment values, both groups exhibited significant improvements in pain score, Constant score, and range of motion measurements at three and six weeks (p<0.05). Pain score decreased by 37.7% in the injection group after 24 hours of injection. Pretreatment Constant scores which were 39.9 + or - 13.9 and 40.3 + or - 13.4 in the injection and control groups increased to 68.4 + or - 7.6 and 64.7 + or - 7.7 at six weeks, respectively. No significant differences were seen in pain scores between the two groups at three and six weeks (p>0.05). Patients receiving subacromial injection had significantly higher Constant scores at six weeks (p=0.044) and significantly greater external and internal rotation at three weeks (p=0.03). Range of motion measurements did not differ between the two groups at six weeks (p>0.05). CONCLUSION: Subacromial injection contributes to the success of the conservative treatment through decreasing pain and enabling more effective range of motion and strengthening exercises, both of which are associated with increased functional improvement. PMID- 19809231 TI - [Use of vacuum-assisted closure in the topical treatment of surgical site infections]. AB - OBJECTIVES: This study was designed to evaluate the results of vacuum-assisted closure in the topical treatment of surgical site infections. METHODS: Vacuum therapy was performed in 17 patients (10 males, 7 females; mean age 60 + or - 20 years) using the VAC system (Vacuum-Assisted Closure, Kinetic Concept Inc) for the treatment of surgical site infections. Infective wounds were in the hip (n=6), crus (n=5), knee (n=3), sacrum (n=2), and hand (n=1). The causative organism for infections was gram-positive bacteria in 15 patients (88.2%), and six patients (35.3%) had nosocomial infections. The wounds were treated with a negative pressure of 100-125 mmHg applied continuously for the first two days, and then intermittently for the following days. The mean follow-up period was 11 + or - 6 months. RESULTS: The mean duration of vacuum therapy was 16 + or - 4 days and the mean length of hospitalization was 31 + or - 19 days. The patients underwent a mean number of three surgical procedures (range 1 to 6) before vacuum therapy. The mean amount of discharge from the wound was 500 + or - 150 ml. Tissue edema and discharge problems were resolved in all the wounds and a hygienic and dry-looking surgical site was attained. In 11 patients (64.7%), clinical and bacteriologic eradication of infections was achieved at the surgical site through antibiotic use and vacuum therapy. Six patients with nosocomial infections continued to receive antibiotic treatment following resolution of surgical site problems. The mean wound area showed a significant reduction from 36 + or - 14 cm(2) to 11 + or - 10 cm(2) following vacuum therapy (p<0.05). Only two patients (11.8%) required further surgical interventions for the closure of wound site. Two patients (11.8%) complained of pain associated with vacuum application. CONCLUSION: Besides its topical advantages in the care of infected wounds, vacuum-assisted closure provides a more rapid and comfortable treatment opportunity, representing a reliable alternative to conventional wound care methods. PMID- 19809232 TI - [The effect of group exercise on postmenopausal osteoporosis and osteopenia]. AB - OBJECTIVES: We investigated the effects of group exercise on bone mineral density (BMD), pain, and quality of life in postmenopausal women with osteoporosis and osteopenia. METHODS: The study included 16 osteoporotic (mean age 55.2 years) and 17 osteopenic (mean age 55.4 years) postmenopausal women whose diagnoses were made by dual energy X-ray absorptiometry (DEXA) showing T-scores of less than 2.5 and in a range of -1 to -2.5, respectively. Subjects having orthopedic, neurological, respiratory, vascular, metabolic, or mental problems were excluded. Each group received the same group exercise program for one hour three times a week for 21 weeks, supervised by a physiotherapist, and including breathing, warm up, stretching, strengthening, balance, stabilization, and cooling exercises. All participants were evaluated before and after the exercise program by a visual analog scale for pain severity, by DEXA for BMD, and by QUALEFFO-41 (Quality of Life Questionnaire of the European Foundation for Osteoporosis) for quality of life. RESULTS: The two groups were similar with respect to age, height, and body mass index (p>0.05), but osteopenic women had a higher body weight (p<0.05). After the exercise program, both groups exhibited significant improvements in T score, pain score, BMD, and all parameters of the QUALEFFO-41 (p<0.05). The mean T-scores before and after exercise were -2.7 + or - 0.2 and -2.4 + or - 0.5 in osteoporotic women, and -1.8 + or - 0.5 and -1.4 + or - 0.5 in osteopenic women, respectively. Following exercise, 43.8% of osteoporotic women had a T-score showing osteopenia, and 23.5% of osteopenic women had a T-score falling within the normal range. The two groups did not differ significantly with respect to the differences between the mean improvements obtained after the exercise program (p>0.05). CONCLUSION: This pilot study demonstrates the effectiveness of physiotherapist-supervised group exercise programs in decreasing pain and increasing BMD and quality of life of both osteoporotic and osteopenic women. PMID- 19809233 TI - [A review of malpractice claims concerning orthopedic applications submitted to the Council of Forensic Medicine]. AB - OBJECTIVES: The aim of this study was to review malpractice claims concerning orthopedic applications that were sent to the Council of Forensic Medicine by public prosecutors and law courts. METHODS: A retrospective review was conducted on 174 malpractice claim files related with orthopedic applications, which had been examined and concluded by the Third Specialized Board of the Council of Forensic Medicine between January 2004 and December 2007. RESULTS: Of 174 files, 129 files were concerned with trauma and 45 files were concerned with orthopedic causes. The types of institutions involved were 82 state hospitals, 56 private hospitals, 17 training and research hospitals, 18 university hospitals, and one military hospital. Orthopedic surgeons were found liable for failure in 61 cases (51 trauma, 10 orthopedic cases). Those who were considered blameworthy were working for 26 private hospitals, 24 state hospitals, seven training and research hospitals, and two university hospitals with the following titles: professor (n=2), chief of clinic (n=1), associate professor (n=2), specialist (n=54), resident (n=3), and nurse (n=1). CONCLUSION: In order to minimize malpractice claims, physicians should steadily improve professional knowledge and skills, give special attention to documenting all information about the patients and applications, and establish a good and intimate physician-patient relationship. PMID- 19809234 TI - [A comparison between three irrigation methods in the debridement of contaminated bovine cancellous bone and the effect of duration of irrigation on the efficiency of debridement]. AB - OBJECTIVES: Irrigation and debridement constitute an important part of treatment of open fractures. We investigated the efficiency of different irrigation methods and durations in cleansing contaminated bovine femur cancellous bone samples and the extent of tissue damage associated with irrigation. METHODS: A total of 72 samples of 4x4x1 cm size were obtained from fresh frozen bovine distal femoral cancellous bone. The top surface of the samples were sawed to a 2-mm depth to create four squares equal in size. All the samples were contaminated with construction sand using the same method and were then randomized to three irrigation groups (bulb syringe irrigation, high-pressure pulsatile lavage, and low-pressure pulsatile lavage), each consisting of 24 samples. The duration of irrigation was set as 3, 6, or 9 minutes for every eight samples of each group. After the irrigation procedure, the images were transferred to a computer screen with a video-microscope camera and the number of sand particles on the samples were counted and irrigation-related macroscopic bone damage was assessed. RESULTS: The lowest number of sand particles was found on the samples irrigated by high-pressure pulsatile lavage (p<0.001). The duration of irrigation did not affect the efficiency of cleansing in all the groups (p>0.05). The least irrigation-related bone damage was observed in samples irrigated by low-pressure pulsatile lavage for 3 minutes (p<0.01). The amount of bone damage was similar in all groups after irrigations beyond 3 minutes (p>0.05). CONCLUSION: Our findings showed that the most efficient method of cleansing contaminated bone samples was high-pressure pulsatile lavage and that prolonged irrigations did not enhance the efficiency of the irrigation method; however, high-pressure irrigation of 3 minute duration resulted in the greatest bone damage. PMID- 19809235 TI - [Evaluation of soft tissue reactions to three nonabsorbable suture materials in a rabbit model]. AB - OBJECTIVES: We evaluated the inflammatory reactions induced by three commonly used nonabsorbable suture materials in a rabbit model. METHODS: Three suture materials were tested: braided polyester suture (Ethibond), braided blend of polyester and polyethylene suture (FiberWire), and monofilament polypropylene suture (Polypropylene). Thirty-six rabbits were randomly allocated to three suture groups, equal in number. Each suture type was placed bilaterally in the quadriceps muscle, patellar tendon, knee joint capsule, and Achilles tendon. Six animals in each group were sacrificed in the third and sixth weeks. The inflammation induced by each suture was assessed using light microscopy and the width of the inflammation zone (WIZ) was measured. RESULTS: Ethibond was found to cause the most severe reaction in the muscle and tendon in the third week; in the sixth week, however, it showed the lowest inflammatory reaction in all tissue types. Reaction to Propylene was moderate in the third week, whereas it caused the largest WIZ in all tissue types in the sixth week, such that the eventual size of the WIZ induced by Propylene (6.6 + or - 2.1 mm) was significantly greater than that of Ethibond (1.6 + or - 0.9 mm) in muscle specimens (p<0.05). Except for the largest WIZ seen in joint capsule specimens in the third week, inflammatory reactions associated with FiberWire were low or moderate in all tissue specimens throughout the study. FiberWire was associated with some necrotic areas in two muscle and one tendon specimens. CONCLUSION: The extent of inflammatory reaction to nonabsorbable suture materials depends on the type of suture material, tissue type, and the duration of postoperative time. PMID- 19809236 TI - [Osteoid osteoma of the coronoid process causing flexion contracture of the elbow]. AB - Osteoid osteoma is one of the benign osteoblastic lesions that causes chronic pain. Diagnosis may be delayed in juxta-articular lesions in which characteristic radiographic findings may not be present, resulting in limited joint motion. A 23 year-old patient presented with a complaint of pain in the right elbow of one year history. He sought medical treatment at another center and was prescribed non-steroidal anti-inflammatory treatment that resulted in significant pain relief; however, limitations of elbow motion ensued. On physical examination, there was pain in the antecubital part of the right elbow on palpation and a flexion contracture of 30 degrees. Findings of computed tomography and magnetic resonance imaging were consistent with osteoid osteoma. The lesion was excised and postoperative controls showed no flexion contracture. PMID- 19809237 TI - [Upper extremity deep vein thrombosis following soft tissue trauma]. AB - A 27-year-old man sustained soft tissue injury to the left shoulder following a fall. Three days later, a diagnosis of left proximal subclavian vein thrombosis was made. The patient was given intravenous urokinase for 24 hours, followed by intravenous unfractionated heparin and oral warfarin. Oral anticoagulant treatment was continued for 12 months. His symptoms completely disappeared after three months of treatment. It is important to be aware of this unusual but potentially serious complication, as early diagnosis and treatment may limit morbidity and mortality. PMID- 19809238 TI - [Intraosseous ganglions at the same localization in twin sisters]. AB - Among radiolucent lesions of the carpal bones, intraosseous ganglions are the most frequent. Most cases involve the lunate or scaphoid bones in the wrist. Two twin sisters aged 23 years were examined for the complaint of left-sided wrist pain. Physical examination and radiologic findings showed unilateral and symmetrical lesions in the left scaphoid bone suggesting intraosseous ganglions. Both patients underwent ganglion excision and grafting using autografts taken from the ipsilateral distal radius. Both patients were free of wrist pain during a three-year follow-up. Histopathologic examination of the excised lesions was consistent with intraosseous ganglion. Unilateral and symmetrical involvement of the same bone in these twin sisters suggests genetic transmission, which has not been previously reported for intraosseous ganglions. PMID- 19809239 TI - Development and characterization of new microsatellite markers for the oyster mushroom (Pleurotus ostreatus). AB - We developed and characterized 36 polymorphic microsatellite markers for the oyster mushroom (Pleurotus ostreatus). In total, 169 alleles were identified with an average of 4.7 alleles per locus. Values for observed (HO) and expected (HE) heterozygosities ranged from 0.027 to 0.946 and from 0.027 to 0.810, respectively. Nineteen loci deviated from Hardy-Weinberg equilibrium. Significant (P<0.05) excess heterozygosity was observed at nine loci. Linkage disequilibrium (LD) was significant (P<0.05) between pairs of locus alleles. Cluster analysis revealed that five species of genus Pleurotus made a distinct group, and the individual cultivars were grouped into major five groups from G-1 to G-5. The diverse cultivars of P. ostreatus were discriminated and the other four species revealed a different section in the UPGMA tree. These microsatellite markers proved to be very useful tools for genetic studies, including assessment of the diversity and population structure of P. ostreatus. PMID- 19809240 TI - Abundance and structure of microbial loop components (bacteria and protists) in lakes of different trophic status. AB - The abundance, biomass, size distribution, and taxonomic composition of bacterial and protistan (heterotrophic and autotrophic nanoflagellates and ciliates) communities were investigated in six lakes of Masurian Lake District (north eastern Poland) differing in trophic state. Samples were taken from the trophogenic water layer during summer stratification periods in the years of 2004 and 2006. Image analysis techniques with fluorescent in situ hybridization (FISH) as well as [3H]-methyl-thymidine incorporation methods were applied to analyze differences in the composition and activity of bacterial communities. The greatest differences in trophic parameters were found between the humic lake and remaining non-humic ones. The same bacterial and heterotrophic nanoflagellate (HNF) cell size classes dominated in all the studied lakes. However, distinct increases in the contributions of large bacterial (>1.0 microm) and HNF (>10 microm) cells were observed in eutrophic lakes. The bacterial community was dominated by the beta-Proteobacteria group, which accounted for 27% of total DAPI counts. Ciliate communities were largely composed of Oligotrichida. Positive correlations between bacteria and protists, as well as between nanoflagellates (both heterotrophic and autotrophic) and ciliates, suggest that concentrations of food sources may be important in determining the abundance of protists in the studied lakes. PMID- 19809241 TI - Combination strategy to increase cyclosporin A productivity by Tolypocladium niveum using random mutagenesis and protoplast transformation. AB - The cyclic undecapeptide cyclosporin A (CyA), one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex by the filamentous fungus Tolypocladium niveum. To increase CyA productivity by wild-type T. niveum (ATCC 34921), random mutagenesis was first performed using an antifungal agar-plug colony assay (APCA) selection approach. This generated a mutant strain producing more than 9-fold greater CyA than the wild-type strain. Additionally, a foreign bacterial gene, Vitreoscilla hemoglobin gene (VHb), was transformed via protoplast regeneration and its transcription was confirmed by RT-PCR in the UV-irradiated mutant cell. This led to an additional 33.5% increase of CyA production. Although most protoplastregenerated T. niveum transformants tend to lose CyA productivity, the optimized combination of random mutagenesis and protoplast transformation described here should be an efficient strategy to generate a commercially valuable, yet metabolite low-producing, fungal species, such as CyA producing T. niveum. PMID- 19809242 TI - Novel alkali-stable, cellulase-free xylanase from deep-sea Kocuria sp. Mn22. AB - A novel xylanase gene, Kxyn, was cloned from Kocuria sp. Mn22, a bacteria isolated from the deep sea of the east Pacific. Kxyn consists of 1,170 bp and encodes a protein of 390 amino acids that shows the highest identity (63%) with a xylanase from Thermobifida fusca YX. The mature protein with a molecular mass of approximately 40 kDa was expressed in Escherichia coli BL21 (DE3). The recombinant Kxyn displayed its maximum activity at 55 degrees and at pH 8.5. The Km, Vmax, and kcat values of Kxyn for birchwood xylan were 5.4 mg/ml, 272 micromol/min.mg, and 185.1/s, respectively. Kxyn hydrolyzed birchwood xylan to produce xylobiose and xylotriose as the predominant products. The activity of Kxyn was not affected by Ca2+, Mg2+, Na+, K+, beta- mercaptoethanol, DTT, or SDS, but was strongly inhibited by Hg2+, Cu2+, Zn2+, and Pb2+. It was stable over a wide pH range, retaining more than 80% activity after overnight incubation at pH 7.5-12. Kxyn is a cellulase-free xylanase. Therefore, these properties make it a candidate for various industrial applications. PMID- 19809243 TI - Isolation and characterization of the eicosapentaenoic acid biosynthesis gene cluster from Shewanella sp. BR-2. AB - Forty-four eicosapentaenoic acid (EPA)-producing microbial strains were isolated from the intestines of marine fishes. Among them, one strain showing a maximum level of EPA (4.78%of total fatty acids) was identified as Shewanella sp. BR-2 on the basis of its 16S rRNA sequence. The EPA content reached a maximum level during the mid-exponential phase of cell growth, and gradually decreased with further growth of the cells. A cosmid DNA including the EPA biosynthesis gene cluster consisting of pfaA-E was isolated from a cosmid library of genomic DNA of Shewanella sp. BR-2, named pCosEPA-BR2. An E. coli clone harboring pCosEPA-BR2 produced EPA at a maximum level of 7.5%of total fatty acids, confirming the EPA biosynthesis activity of the cloned gene cluster. PMID- 19809244 TI - Lipase diversity in glacier soil based on analysis of metagenomic DNA fragments and cell culture. AB - Lipase diversity in glacier soil was assessed by culture independent metagenomic DNA fragment screening and confirmed by cell culture experiments. A set of degenerate PCR primers specific for lipases of the hormone-sensitive lipase family was designed based on conserved motifs and used to directly PCR amplify metagenomic DNA from glacier soil. These products were used to construct a lipase fragment clone library. Among the 300 clones sequenced for the analysis, 201 clones encoding partial lipases shared 51-82%identity to known lipases in GenBank. Based on a phylogenetic analysis, five divergent clusters were established, one of which may represent a previously unidentified lipase subfamily. In the culture study, 11 lipase-producing bacteria were selectively isolated and characterized by 16S rDNA sequences. Using the above mentioned degenerate primers, seven lipase gene fragments were cloned, but not all of them could be accounted for by the clones in the library. Two full-length lipase genes obtained by TAIL-PCR were expressed in Pichia pastoris and characterized. Both were authentic lipases with optimum temperatures of or =2 cm showed independent influence on the rate of lymph node metastasis. CONCLUSIONS: Endoscopic mucosectomy in m3 carcinoma is questionable and in all submucosal carcinomas and lesions > or =2 cm it is not indicated. PMID- 19809299 TI - Biliary reconstruction using a side-to-side choledochocholedochostomy with or without T-tube in deceased donor liver transplantation: a prospective randomized trial. AB - OBJECTIVE: The biliary anastomosis is still one of the major causes for morbidity after orthotopic liver transplantation. The optimal method of reconstruction remains controversial. The aim of the study was to assess biliary complications after liver transplantation using a choledochocholedochostomy with or without a temporary T-tube. BACKGROUND DATA: Several reports have suggested that biliary reconstruction without T-tube is a safer method with a lower rate of biliary complications compared with T-tube insertion. METHODS: A total of 194 recipients of deceased donor liver grafts were randomized. In group 1 the biliary reconstruction was performed by side-to-side choledochocholedochostomy with (n = 99) and in group 2 (n = 95) without a T-tube. The T-tube was removed after 6 weeks. RESULTS: The overall biliary complication rate was significantly increased in group 2 (P < 0.0005). Biliary leaks occurred in 5 patients in group 1 and in 9 patients in group 2 (5.05% vs. 9.47%; P = 0.2756 ns). Anastomotic strictures of the bile duct were seen in 7 patients in group 1 and in 8 patients in group 2 (7.07% vs. 8.42%; P = 0.7923 ns). Two of the patients in group 1 and 5 patients in group 2 developed an ischemic type biliary lesion (2.02% vs. 5.26%; P = 0.2716 ns). The rate of reoperations was comparable in both groups. The rate of invasive interventions was higher in the group without T-tubes (9% vs. 18%, P = ns), as was the rate of cholangitis (5% vs. 11%. P = ns) and pancreatitis (4% vs. 14%, P = 0.0218). No complications after removal of the T-tube were observed. CONCLUSION: This study is a large prospective randomized trial to assess biliary complications that occur following liver transplantation, after anatomizing the bile duct with or without T-tubes. A significant increased rate of complications in the group without T-tube insertion was observed. In summary, our results indicate that the usage of T-tubes is safe and an excellent tool for the quality control of biliary anastomoses. PMID- 19809300 TI - Cellular cytotoxicity and in-vivo biodistribution of docetaxel poly(lactide-co glycolide) nanoparticles. AB - Docetaxel (DTX) is one of the most effective antineoplastic drugs. However, its current clinical administration, formulated in tween80, causes serious side effects. This study is focused on preparation and evaluation of poly(lactide-co glycolide) nanoparticles (NPs) containing DTX to remove tween80. Drug encapsulation efficiency, in-vitro drug release, cellular cytotoxicity, and in vivo biodistribution of NPs in mice after intravenous administration were investigated. The average diameter of the NPs was approximately 172-178 nm with encapsulation efficiency of 68%. A burst release of approximately 30% (w/w) of the loaded drug followed by a sustained release profile was observed. Cellular mortality of the NPs was more than or at least as great as DTX free drug; for example, cell viability measured at 100 nmol/l drug concentration was decreased from 50.9% for DTX free drug to 15.9% for the NP formulation after 48 h incubation with T47D cells. The DTX plasma amount remained at a good level (13% of the initial dose) in the NP formulation compared with the DTX conventional formulation, which is approximately 0.5% of the initial dose, was present in plasma up to 2 h. Poly(lactide-co-glycolide) NPs containing DTX prepared in this study may be regarded as a suitable and superior formulation for the current formulation in the market containing tween80 with improved cancerous cell mortality and biodistribution characteristics. PMID- 19809301 TI - Hybrid capture-2 is better than in situ hybridization: a study comparison with cervical biopsies. PMID- 19809302 TI - Diffuse membranous immunoreactivity of CD56 and paranuclear dot-like staining pattern of cytokeratins AE1/3, CAM5.2, and OSCAR in undifferentiated (embryonal) sarcoma of the liver. PMID- 19809303 TI - Identification and prevalence of von Willebrand disease type 2N (Normandy) in Australia. AB - We report an investigation of type 2N von Willebrand disease (VWD), covering the past 7 years and evaluating 1031 plasma samples from over 500 patients. Samples included specific requests for investigation of possible type 2N VWD (including family studies) and samples from 'hemophilia' or nonspecified VWD investigations that could unknowingly be type 2N VWD. In total, 13 new patients with type 2N VWD were identified, four of whom initially presented with normal levels of factor VIII and only three of whom (i.e. 23%) derived from specific clinical requests for investigation of type 2N VWD. Furthermore, type 2N VWD was excluded in 91% of specific clinical requests for type 2N VWD investigations. Poststudy evaluation indicates that type 2N VWD in this geographic region has an incidence rate similar to that in other westernized regions, accounting for around 1-2% of all identified VWD cases and about 13% of all type 2 VWD cases. In conclusion, this study highlights that clinicians requesting laboratory investigations related to a bleeding tendency often fail to appropriately recognize the possibility of type 2N VWD, that a normal plasma factor VIII will not exclude type 2N VWD, and although a relatively uncommon form of VWD overall, type 2N VWD represents a significant qualitative disorder. PMID- 19809304 TI - Analysis of plasmin generation and clot lysis of plasma fibrinogen purified from a heterozygous dysfibrinogenemia, BbetaGly15Cys (Hamamatsu II). AB - We found a heterozygous dysfibrinogenemia caused by the substitution of BbetaGly15Cys and designated it fibrinogen Hamamatsu II (H-II). Although the propositus suffered an infarction of the medulla oblongata, other thrombotic risk factors, paradoxical cerebral infarction, and arterial dissection were not found. To determine whether the delayed lysis of fibrin clots or not in the context of the BbetaGly15Cys substitution, we examined the clot lysis and plasmin generation of propositus' fibrinogen. Fibrinogen was purified from the propositus' and normal control plasma by immunoaffinity chromatography and was used for the following experiments: sodium dodecyl sulfate-polyacrylamide gel electrophoresis, fibrin polymerization, scanning electron microscopic observation of fibrin clot and fibers, clot lysis, and tissue-type plasminogen activator-mediated plasminogen activation. The H-II plasma fibrinogen showed the presence of albumin binding variant forms, a dimeric molecule of variant fibrinogen, and impairment of lateral aggregation during fibrin polymerization. The H-II fibrin clot showed lower density of bundles and thinner diameters of fibers than in the normal fibrin clot. In the clot lysis experiments with overlaid plasmin, H-II fibrin showed a similar lysis period and lysis rate to the normal control. Moreover, plasmin generation from a mixture of thrombin, tissue-type plasminogen activator, plasminogen, and H-II fibrinogen also showed a similar rate to normal fibrinogen. Although the propositus suffered an infarction, the present study did not observe delayed clot lysis, that is, the clot was not resistant to plasmin degradation. Therefore, we did not clarify an association between the BbetaGly15Cys dysfibrinogenemia and arterial thrombosis. PMID- 19809305 TI - Activated factor XII type A and B-type natriuretic peptide are complementary and incremental predictors of mortality in patients following admission with acute coronary syndrome. AB - The aim of this analysis was to assess the predictive value of activated factor XII type A (XIIaA) and B-type natriuretic peptide (BNP) in acute coronary syndrome patients stratified according to troponin release and to evaluate their complementary utility as predictors of all-cause mortality and recurrent troponin T (TnT)-positive events. Multivariable analysis in 870 patients admitted with suspected myocardial infarction was performed using the Cox proportional hazard ratio model. Variables in the model included XIIaA and BNP as well as conventional risk factors for mortality. Although both XIIaA and BNP were identified as independent predictors for all-cause mortality in the total group of patients, only BNP was found to be an independent predictor for all-cause mortality in patients with a confirmed myocardial infarction (TnT > 0.05 ng/ml) at admission (hazard ratio 4.24, 95% confidence interval 1.28-14.07), whereas only XIIaA was an independent predictor for all-cause mortality in patients with low TnT release (0.01 < TnT < or = 0.05 ng/ml) at admission (hazard ratio 10.37, 95% confidence interval 2.89-37.21). The combination of these two biomarkers provided complementary prognostic information for all-cause mortality as compared with each of the biomarkers alone in the total patient material. XIIaA is particularly useful in predicting mortality in acute coronary syndrome patients with low troponin release, whereas BNP is effective in predicting mortality in patients with confirmed myocardial infarction and more substantial troponin release. The combination of these two biomarkers improves outcome prediction in unselected patients with chest pain and acute coronary syndrome. PMID- 19809306 TI - Plasma thrombomodulin activity, tissue factor activity and high levels of circulating procoagulant phospholipid as prognostic factors for acute myocardial infarction. AB - Several studies have indicated an association between haemostatic markers and acute myocardial infarction, but few or no studies refer to their activity. We studied plasma levels of 10 coagulation factors (fibrinogen, protein C, protein S, von Willebrand factor, D-dimers, factor VIIa, free tissue factor pathway inhibitor, tissue-type plasminogen activator, plasminogen activator inhibitor-1, thrombomodulin) and using new specific assays analysed the activity of plasma tissue factor (TFa), thrombomodulin (TMa), and procoagulant phospholipid in 46 consecutive patients with acute myocardial infarction at the time of hospital admission, and compared them with 34 healthy normal volunteers. Plasma levels of TFa, TMa, and procoagulant phospholipid were significantly higher in cases than in control patients (P < 0.001). In addition the ratio of TFa/free tissue factor pathway inhibitor was higher in patients than in controls, whereas the tissue type plasminogen activator (t-PA)/plasminogen activator inhibitor-1 ratio was lower in patients. Interestingly, patients with an unfavourable outcome during a 2-month follow-up had higher levels of TFa, TMa, procoagulant phospholipid, a higher ratio of TFa/free tissue factor pathway inhibitor and a lower ratio of t PA/plasminogen activator inhibitor-1 than patients who recovered. The combination of these different parameters reveals an increase in procoagulant activity as well as impaired fibrinolytic activity during the acute phase of an acute myocardial infarction. The association of the level of the activity of these three factors may provide a new tool to assess the prognosis of acute myocardial infarction. Further studies are needed to support our findings and to elucidate the clinical interest of measuring these factors. PMID- 19809307 TI - The fibrinolytic mechanism of defibrotide: effect of defibrotide on plasmin activity. AB - Fibrinolytic activity has been shown to be reduced in many vascular diseases, including hepatic veno-occlusive disease after stem cell transplantation, a microangiopathy characterized by sinusoidal endothelial cell injury. Defibrotide is a polydisperse oligonucleotide with antithrombotic, profibrinolytic, anti ischemic, and antiadhesive properties. Numerous clinical studies have shown promising activity of defibrotide in the treatment and prevention of veno occlusive disease, with minimal toxicity. In corollary laboratory studies, defibrotide has been shown to decrease plasminogen activator inhibitor-1, increase tissue plasminogen activator levels, and increase overall plasma fibrinolytic activity in patients. Plasmin, a potent and nonspecific serine protease, plays a pivotal role in fibrinolysis by virtue of its ability to effectively degrade fibrin clots. In this study, defibrotide increases the activity of plasmin in hydrolyzing its substrate in a dose-dependent and length dependent manner. Similar concentration-dependent effects of defibrotide were observed when plasmin was generated by tissue plasminogen activator or urokinase activation of plasminogen. In contrast, defibrotide had no direct effect on the activation of plasminogen to plasmin. Defibrotide was also able to enhance the activity of plasmin in degrading fibrin clot formed from fibrinogen, plasminogen, and thrombin. This effect was also concentration-dependent and directly correlated with the enzymatic activity of plasmin. This study therefore demonstrates that defibrotide is capable of enhancing the activity of plasmin and so contributes to its fibrinolytic activity. Taken together, these results support the effect of defibrotide in restoring the fibrinolytic vascular phenotype, in microangiopathic conditions such as veno-occlusive disease. PMID- 19809308 TI - ADAMTS13 and von Willebrand factor concentrations in patients with diabetes mellitus. AB - The von Willebrand factor (VWF) is elevated in patients with diabetes mellitus and degraded by a metalloprotease, ADAMTS13. We hypothesized that this elevation is due to a decreased function of ADAMTS13. Thus, we investigated ADAMTS13 in patients with diabetes mellitus without and with peripheral artery occlusive disease (PAOD). When treating the latter group with dalteparin, VWF is reported to increase significantly, and we therefore measured ADAMTS13 also in these patients. VWF antigen and ADAMTS13 antigen and activity concentrations were measured in patients with diabetes mellitus but without PAOD (diabetes mellitus; n = 23) and with diabetes mellitus and PAOD (diabetes mellitus + PAOD; n = 65) before and after treatment with dalteparin or placebo. In the diabetes mellitus group, concentration of VWF antigen was significantly higher, whereas that of ADAMTS13 activity was significantly lower than in the healthy controls. In the diabetes mellitus along with PAOD group, VWF antigen was significantly higher, but ADAMTS13 antigen or activity did not differ significantly from those of healthy controls. The ADAMTS13 activity/antigen ratio was lower than in controls only in the diabetes mellitus patient group. VWF antigen increased significantly during dalteraprin treatment, whereas ADAMTS13 activity and antigen remained unchanged. Only patients with diabetes mellitus had significantly lower concentrations of ADAMTS13 activity in plasma than controls, although the diabetes mellitus along with PAOD had a more pronounced VWF antigen elevation than diabetes mellitus patients, illustrating a possible link between ADAMTS13 and microangiopathy in diabetes mellitus patients. The increase in VWF antigen concentration during treatment with dalteparin does not seem to be due to changes in ADAMTS13. PMID- 19809309 TI - Anticoagulation after liver transplantation: a retrospective audit and case control study. AB - Anticoagulation may in the future become a therapeutic option for the prevention of liver fibrosis, such as due to recurrent hepatitis C virus infection after liver transplantation. Currently, there are other indications for anticoagulation after liver transplantation but no data regarding its safety. The objective of the study was to audit the safety of anticoagulation after liver transplantation. Liver transplant recipients receiving anticoagulation postoperatively were compared with a matched control group with respect to bleeding complications and postoperative course. Anticoagulation did not increase the risk of bleeding complications after liver transplantation. On the basis of safety, it appears feasible to use anticoagulation in trials to assess prevention of liver fibrosis. PMID- 19809310 TI - Complex aortic arch aneurysm and dissections: hybrid techniques for surgical and endovascular therapy. AB - PURPOSE OF REVIEW: Open aortic arch surgery is highly invasive and may result in significant morbidity. Recent innovations in thoracic stent-graft technology have facilitated methods of replacing the aortic arch without deep hypothermic circulatory arrest or even cardiopulmonary bypass. We present a review of the current techniques and results of hybrid aortic arch surgery centered around a new classification scheme for these procedures. RECENT FINDINGS: Hybrid arch techniques have been developed to address various arch disorders, including aneurysmal disease and aortic dissections. In some cases, new proximal or distal landing zones need to be created for adequate seal. Preliminary results show that hybrid arch procedures are potentially beneficial versus open surgery in elderly, very high-risk patients but that this advantage may not be as pronounced in younger lower risk patients. Although still highly experimental, totally endoscopic approaches to aortic arch surgery have been performed successfully. SUMMARY: Hybrid approaches to aortic arch disorder continue to evolve as techniques and technology are further refined. Further long-term outcome data on these procedures will delineate their role in the future. PMID- 19809311 TI - The role of low endothelial shear stress in the conversion of atherosclerotic lesions from stable to unstable plaque. AB - PURPOSE OF REVIEW: Local hemodynamic factors are major determinants of the natural history of individual atherosclerotic plaque progression in coronary arteries. The purpose of this review is to summarize the role of low endothelial shear stress (ESS) in the transition of early, stable plaques to high-risk atherosclerotic lesions. RECENT FINDINGS: Low ESS regulates multiple pathways within the atherosclerotic lesion, resulting in intense vascular inflammation, progressive lipid accumulation, and formation and expansion of a necrotic core. Upregulation of matrix-degrading proteases promotes thinning of the fibrous cap, severe internal elastic lamina fragmentation, and extracellular matrix remodeling. In the setting of plaque-induced changes of the local ESS, coronary regions persistently exposed to very low ESS develop excessive expansive remodeling, which further exacerbates the proinflammatory low ESS stimulus. Recent studies suggest that the effect of recognized cardioprotective medications may be mediated by attenuation of the proinflammatory effect of the low ESS environment in which a plaque develops. SUMMARY: Low ESS determines the severity of vascular inflammation, the status of the extracellular matrix, and the nature of wall remodeling, all of which synergistically promote the transition of stable lesions to thin cap fibroatheromata that may rupture with subsequent formation of an occlusive thrombus and result in an acute coronary syndrome. PMID- 19809312 TI - Neuroadrenergic dysfunction in obesity: an overview of the effects of weight loss. AB - PURPOSE OF REVIEW: The prevalence of obesity is rising to epidemic proportions worldwide, and in tandem so is that of type 2 diabetes. Neuroadrenergic abnormalities, comprising increased resting sympathetic nervous system activity and blunted sympathetic neural responsiveness are recognized features of metabolic syndrome obesity, which contribute importantly to both the pathophysiology and adverse clinical prognosis of this high-risk population. Weight loss is recommended as first-line treatment for obesity. This review examines the effects of nonpharmacological weight loss on sympathetic nervous system function under basal and stimulated conditions. RECENT FINDINGS: Human weight loss trials show that even moderate weight reduction is accompanied by significant attenuation in resting whole-body norepinephrine spillover rate and muscle sympathetic nerve activity, an improvement in cardiac autonomic modulation, and a reversal of blunted sympathetic responsiveness at both peripheral and central nervous system levels. Recent findings underscore the relevance of insulin resistance in mediating blunted sympathetic responsiveness to endogenous hyperinsulinemia induced by glucose ingestion. Impaired insulin transport across the blood-brain barrier may be one mechanism mediating these effects. Weight loss reverses blunted sympathetic responsiveness to glucose, which has implications for postprandial energy expenditure and body weight homeostasis. SUMMARY: The autonomic dysfunction of obesity is reversible with weight loss, highlighting the importance of lifestyle intervention as a key therapeutic modality. PMID- 19809313 TI - Genetic variants of the FADS1 FADS2 gene cluster as related to essential fatty acid metabolism. AB - PURPOSE OF REVIEW: The delta-5 and delta-6 desaturases have long been known to be important enzymes in the endogenous formation of long-chain polyunsaturated fatty acids (LC-PUFAs). Cloning of the coding sequences and chromosomal localization of the desaturase encoding genes fatty acid desaturase 1 and 2 (FADS1 and FADS2) opened the way for analyses of genetic factors as regulators of desaturase activity and LC-PUFA homeostasis. The present review summarizes the recent association studies on FADS genotypes and LC-PUFA levels and suggests ideas how FADS genotypes can be integrated in future research. RECENT FINDINGS: An initial candidate gene study reported highly significant associations between FADS gene cluster polymorphisms and fatty acid levels in serum phospholipids with an extraordinary high genetically explained variance for arachidonic acid levels of 28.5%. Carriers of the minor alleles had enhanced levels of desaturase substrates and decreased levels of desaturase products, suggesting a decline in desaturase expression or activity because of the polymorphisms. These results were replicated in several association studies additionally showing an effect in different human tissues as well as in a recent genome-wide association study on LC-PUFA levels. SUMMARY: The validated strong association between FADS genotypes and fatty acid levels in diverse human tissues shows that FADS gene cluster polymorphisms are, in addition to nutritional regulation of fatty acid synthesis, a very important regulator of LC-PUFA synthesis. PMID- 19809314 TI - Productive symptoms in right brain damage. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize the more recent studies on productive symptoms from the neuropsychological, neurophysiological and anatomical points of view. The integration of these aspects may provide some clarifications on the cognitive impairments underpinning the main productive disorders, also contributing to better understand the normal functioning of the brain. RECENT FINDINGS: Productive symptoms are closely associated to spatial neglect and are distinguished in relation to the part of space they manifest. The investigation of perseveration in extrapersonal space with different manipulations helps to understand the neuropathological mechanisms underlying this symptom. Anosognosia for hemiplegia and somatoparaphrenia may be considered as disorders of body representation (personal space). Recently it has been proposed that these disorders may be ascribed to an impairment of different levels of motor control. The identification of the anatomical correlates of these two disorders contributes to better understanding of their the cognitive nature. SUMMARY: Productive behaviours have diverse clinical manifestations and may be induced by different mechanisms. Lesional studies are beginning to provide evidence for specific anatomical correlates of these disorders. Further investigations are needed to better understand to what extent productive symptoms can be disentagled from spatial neglect. These attempts may contribute to clarifying the role of the right hemisphere in monitoring spatial cognition. PMID- 19809315 TI - Anosognosia: clinical and ethical considerations. AB - PURPOSE OF REVIEW: This review considers recent findings on classical forms of anosognosia and less dramatic disturbances in self-awareness observed in various brain disorders. It also addresses issues of clinical management and ethical treatment. RECENT FINDINGS: The study of anosognosia for hemiplegia, Anton's syndrome, and less dramatic disturbances in self-awareness in conditions such as Alzheimer's disease, mild cognitive impairment, and severe traumatic brain injury suggests its 'multifactorial' nature. Lesions in various regions of the brain may contribute to different forms of anosognosia. Animal and human studies suggest that the insular cortex plays an important role in subjective awareness of feeling states, and may be implicated in various forms of anosognosia. Also, right frontal lobe lesions have been implicated, but typically in patients who have bilateral cerebral dysfunction. These patients require careful clinical management in light of their reduced awareness of their neurological and neuropsychological functions that impact daily activities. SUMMARY: The study of anosognosia and disorders of self-awareness has expanded greatly over the past 20 years. Various patient groups may show different levels of impaired awareness or anosognosia. Guidelines for the clinical management and ethical treatment of these patients are needed, but beginning efforts have been made. PMID- 19809316 TI - Long-term quality of life in gynecological cancer survivors. AB - PURPOSE OF REVIEW: This review addresses current knowledge about long-term quality of life (QOL) in survivors of gynecological cancer. RECENT FINDINGS: Survivors of gynecological cancer have generally good long-term QOL, equivalent to healthy controls; however, specific deficits are more prevalent than in women without cancer. Ovarian cancer survivors have good QOL, with few physical symptoms. Psychological distress and sexual impairments exist. No differences in QOL occur between survivors of early and advanced stage disease. Cervical cancer survivors treated with radiotherapy reported more QOL impairments than survivors treated with other approaches. There were no differences on sexuality and sexual function among survival time periods. In general, cervical cancer survivors seem to have a positive attitude towards sexuality. Self-esteem is an important psychological variable in the study of long-term QOL. SUMMARY: When considering specific diseases, ovarian and cervical cancers were the most researched. Endometrial cancer was underreported in recent literature. Studies addressing vulvar and vaginal cancers are lacking. Physical, psychosocial and sexuality were the most investigated QOL domains. Advances are observed in current research; however, more rigorous and larger studies are required to further understand long term QOL. Available findings are crucial to develop interventions to support those at risk for QOL impairments. PMID- 19809317 TI - New guidelines for weight gain during pregnancy: what obstetrician/gynecologists should know. AB - PURPOSE OF REVIEW: To review the recently issued guidelines for weight gain during pregnancy. RECENT FINDINGS: These guidelines were developed to minimize the negative health consequences for both mother and fetus of inadequate or excessive weight gain. They call for categorizing women's prepregnancy BMI using the WHO/National Heart, Lung and Blood Institute cutoff points and provide ranges of recommended weight gain for underweight (28-40 lb), normal weight (25-35 lb), overweight (15-25 lb) and obese (11-20 lb) gravidas. Data were insufficient to construct specific guidelines for women with class II or class III obesity. Women should attempt to conceive at a normal weight for better obstetric outcomes. Improved comprehensive preconceptional care is necessary to help women reach this goal. Most American women currently gain weight below or above the new ranges, so changes are required in both women's behavior and how their care is managed. Data from a variety of interventions related to improved diet and increased physical activity show that individualized care can assist women in gaining weight within these guidelines. SUMMARY: The guidelines offer many opportunities for obstetrician/gynecologists, together with ancillary healthcare providers, to assume a larger role as 'women's healthcare physicians' and to conduct research that could improve the health of mothers and children. PMID- 19809318 TI - Management strategies for ovulation induction in women with polycystic ovary syndrome and known clomifene citrate resistance. AB - PURPOSE OF REVIEW: Clomifene citrate is the first and the most used agent for inducing ovulation in patients affected by polycystic ovary syndrome (PCOS). About 60-85% of PCOS women ovulated under clomifene citrate, whereas the others were defined clomifene citrate-resistant. The purpose of the current review will be to describe treatment strategies to induce ovulation in infertile PCOS patients with clomifene citrate resistance. RECENT FINDINGS: Clomifene citrate and metformin association are a valid option for inducing ovulation in clomifene citrate-resistant PCOS patients. Surgical ovulation induction by laparoscopic ovarian drilling should be reserved to well selected cases. Excellent preliminary results are obtained using new drug formulations, such as aromatase inhibitors. SUMMARY: In clomifene citrate-resistant PCOS patients, clomifene citrate and metformin combination and laparoscopic ovarian drilling, in selected cases, should be considered before gonadotropin administration. The efficacy of the other treatments must be confirmed in future well designed studies. PMID- 19809319 TI - Multiple sclerosis. Part I: neuro-ophthalmic manifestations. AB - PURPOSE OF REVIEW: This update includes topics relating to multiple sclerosis (MS) for the ophthalmologist. RECENT FINDINGS: Interest in the ophthalmology of MS is highlighted by the substantial amount of original research presented at recent medical conferences. At the 2008 World Congress on Treatment and Research in Multiple Sclerosis, 33 of 898 abstract presentations related to ophthalmology. The most represented topics were optical coherence tomography (OCT) and radiology of the visual system (19/33 posters). The 2009 North American neuro ophthalmologic society (NANOS) conference featured MS-related topics in 6 of 24 platform presentations. This article reviews recent advancements in the neuro ophthalmology of MS. SUMMARY: Multiple sclerosis results in various ophthalmic abnormalities. Optic neuritis is the most widely studied visual disturbance in MS, but other unusual visual manifestations such as Charles Bonnet syndrome or the Pulfrich phenomenon also occur. OCT is used to measure thickness of the retinal nerve fiber layer. Because thinning of this layer in MS represents axonal loss, OCT is being used to evaluate neuroprotective treatments. The efferent visual system, in particular the medial longitudinal fasciculus pathway, may be a way to measure body temperature-induced changes in MS. PMID- 19809320 TI - Pituitary apoplexy: evaluation, management, and prognosis. AB - PURPOSE OF REVIEW: To review the current standard of care in the diagnosis and treatment of pituitary apoplexy and to determine any updated clinical management strategies. RECENT FINDINGS: Pituitary apoplexy is a rare but life-threatening medical emergency. Presenting signs and symptoms often include severe headache, visual loss, ophthalmoplegia, altered consciousness, and impaired pituitary function. Common predisposing factors include closed head trauma, blood pressure alterations, history of pituitary irradiation, cardiac surgery, anticoagulation, treatment with dopamine agonists, pituitary stimulation testing, and pregnancy. MRI imaging is the most sensitive sequence for the detection of acute and old intracranial hemorrhage. Patients often require emergent intravenous fluids, blood transfusions, and high-dose corticosteroids. Patients who remain clinically and neurologically unstable require urgent transsphenoidal surgical decompression as definitive treatment. SUMMARY: In patients with pituitary apoplexy, improvement in visual field defects, visual acuity, and diplopia is typically observed after emergent application of therapy, often including medical and surgical treatment. Some patients may require long-term hormonal therapy after surgery. PMID- 19809321 TI - Current research on affective temperaments. AB - PURPOSE OF REVIEW: The aim of this review is to highlight the relationship between affective temperaments and clinical mood disorders and to summarize the earlier and most recent studies on affective temperaments in both clinical and nonclinical populations. RECENT FINDINGS: Current research findings show that specific affective temperament types (depressive, cyclothymic, hyperthymic, irritable and anxious) are the subsyndromal (trait-related) manifestations and commonly the antecedents of minor and major mood disorders. Up to 20% of the population has some kind of marked affective temperaments; depressive, cyclothymic and anxious temperament is more frequent in women, whereas hyperthymic and irritable temperaments predominate among men. Molecular genetic studies show a strong involvement of the central serotonergic (depressive, cyclothymic, irritable and anxious temperaments) and dopaminergic (hyperthymic temperament) regulation, suggesting that the genetic potential of major mood episodes lies in these temperaments. SUMMARY: Premorbid affective temperament types have an important role in the clinical evolution of minor and major mood episodes including the direction of the polarity and the symptom formation of acute mood episodes. They can also significantly affect the long-term course and outcome including suicidality and other forms of self-destructive behaviours such as substance use and eating disorders. PMID- 19809322 TI - What is neuroethics? Empirical and theoretical neuroethics. AB - PURPOSE OF REVIEW: Neuroethics is a recently emerging field that deals with predominantly empirical and practical issues of ethics in neuroscience. In contrast, theoretical and methodological considerations have rather been neglected and thus what may be called theoretical neuroethics. RECENT FINDINGS: The review focuses on informed consent and moral judgment as examples of empirical neuroethics and norm-fact circularity and method-based neuroethics as issues of a theoretical neuroethics. SUMMARY: It is argued that we need to consider theoretical and methodological issues in order to develop neuroethics as a distinct discipline, which as such can be distinguished from both philosophy/ethics and neuroscience. PMID- 19809323 TI - Effects of increased inspired oxygen concentration on tissue oxygenation: theoretical considerations. AB - BACKGROUND AND OBJECTIVES: Breathing increased fractional oxygen concentration (FiO2) is recommended for the treatment of tissue ischaemia. METHODS: The theoretical benefits of increasing FiO2 on tissue oxygenation were evaluated using standard physiological equations. Assuming constant oxygen consumption by tissues throughout the length of a capillary, the oxygen content at 20 arbitrary points along a capillary was calculated. Using mathematical representations of the haemoglobin dissociation curve and an iterative approach to include the dissolved oxygen component of oxygen content, the oxygen partial pressure (PO2) profile along a capillary was estimated. RESULTS: High FiO2 concentrations cause large increases in PO2 at the arteriolar end of capillaries but these large PO2 values, caused by the extra dissolved oxygen, rapidly decline along the capillary. At the venular end of the capillary (the area of tissue most likely to be hypoxic), breathing oxygen causes only a modest improvement in PO2. CONCLUSION: Increasing FiO2 to treat tissue hypoxia has clear benefits, but a multimodal approach to management is required. PMID- 19809324 TI - Effects of articaine and ropivacaine on calcium handling and contractility in canine ventricular myocardium. AB - BACKGROUND AND OBJECTIVE: In spite of the widespread clinical use of articaine and ropivacaine there is little information available on the effects of these drugs on myocardial Ca handling. In the present study, therefore, the concentration-dependent effects of articaine and ropivacaine on the components of intracellular Ca handling were studied and compared in canine ventricular myocardium. METHODS: Contractility was measured in ventricular trabeculae, [Ca]i transients were recorded from electrically stimulated isolated ventricular myocytes loaded with the calcium-sensitive dye fura-2, L-type Ca current was recorded under whole cell patch clamp conditions, and the release and reuptake of Ca was monitored in sarcoplasmic reticulum vesicles. RESULTS: Articaine and ropivacaine caused a reversible and concentration-dependent decrease in amplitude of the [Ca]i transient (EC50 = 87.4 +/- 12 and 99.3 +/- 17 micromol l, respectively), which was congruent with the reduction obtained for contractility (EC50 = 73.7 +/- 10 and 72.8 +/- 14 micromol l, respectively). No significant change in diastolic [Ca]i was found. L-type Ca current was significantly reduced by articaine and ropivacaine with EC50 values of 327 +/- 56 and 263 +/- 67 micromol l, respectively. Neither Ca release and Ca uptake nor the ATPase activity of the sarcoplasmic reticulum vesicles was altered by articaine or ropivacaine at concentrations less than 200 micromol l. In summary, articaine and ropivacaine caused no significant changes at the therapeutically relevant concentrations of the micromolar range. No significant differences between the effects of articaine and ropivacaine on contractility, [Ca]i transients, L-type Ca current, and sarcoplasmic reticulum Ca release and uptake were observed. CONCLUSIONS: Under conditions of normal application both articaine and ropivacaine are free of cardiodepressant effects; however, a negative inotropic action can be anticipated in cases of accidental intravenous injection or overdose. The observed negative inotropic actions of articaine and ropivacaine are similar in magnitude, and can be mainly attributed to a reduction in net trans-sarcolemmal Ca influx. PMID- 19809325 TI - Introduction of bispectral index monitoring in a district general hospital operating suite: a prospective audit of clinical and economic effects. AB - BACKGROUND AND OBJECTIVE: Randomized controlled trials have shown several beneficial effects of intraoperative bispectral index monitoring. We conducted a prospective audit to determine whether these could be replicated in everyday clinical practice. METHODS: Recovery characteristics and drug costs from two 4 week periods were compared, immediately before (phase 1, n = 427 patients) and after (phase 2, n = 299 patients) the introduction of bispectral index monitoring in the main operating theatres of a district general hospital. RESULTS: Demographic and intraoperative variables for the two patient groups were similar. Nausea (P = 0.002), vomiting (P = 0.008) and antiemetic use (P = 0.001) in the postanaesthesia care unit all decreased in phase 2, though unrelated changes in antiemetic policy may have been partly responsible. Recovery time was unaffected. Drug costs decreased in phase 2 by an average of 0.86 pounds per patient. The cost of each semi-reusable sensor was 6.60 pounds. CONCLUSION: In a prospective audit, benefits of intraoperative bispectral index monitoring in our district general hospital were not seen to the same degree as in randomized controlled trials elsewhere. PMID- 19809326 TI - Prophylactic amiodarone versus lidocaine for prevention of reperfusion ventricular fibrillation after release of aortic cross-clamp. AB - BACKGROUND AND OBJECTIVE: Ventricular fibrillation is common after aortic cross clamp release in patients undergoing open-heart surgeries. The aim of the study was to evaluate the efficacy of the prophylactic administration of 150 mg amiodarone by way of the pump 2 min before release of aortic cross-clamp in preventing ventricular fibrillation. METHODS: The present study is a prospective, randomized, controlled and blinded study performed at a teaching university hospital where 120 patients undergoing coronary bypass graft surgery were randomly assigned to three groups. Each group received either 150 mg of amiodarone or 100 mg lidocaine or isotonic saline by way of pump 2 min before release of the aortic cross-clamp. The frequency of occurrence of ventricular fibrillation and the subsequent required defibrillation counter shocks were determined in all groups. RESULTS: The frequency of occurrence of ventricular fibrillation was significantly higher in both the amiodarone (48%) and the control group (45%) as compared with the lidocaine group (20%) with no statistically significant difference between the amiodarone and the control groups. Furthermore, when ventricular fibrillation occurred, the percentage of patients requiring defibrillation counter shocks was significantly higher in both the amiodarone (58%) and control (61%) groups as compared with the lidocaine group (13%) with no difference between the amiodarone and the control groups, despite a significant decrease in the defibrillation counter shocks energy requirements in the amiodarone group. CONCLUSION: The present study showed no difference between amiodarone (150 mg) and placebo in preventing ventricular fibrillation after release of aortic cross-clamp. In addition, the use of lidocaine was able to reduce the incidence of ventricular fibrillation as compared with both amiodarone and placebo. PMID- 19809327 TI - A novel technique for learning to intubate with the lightwand. PMID- 19809328 TI - Management of the predicted difficult airway: a comparison of conventional blade laryngoscopy with video-assisted blade laryngoscopy and the GlideScope. AB - BACKGROUND AND OBJECTIVE: We investigated whether the use of two different video laryngoscopes [direct-coupled interface (DCI) video laryngoscope and GlideScope] may improve laryngoscopic view and intubation success compared with the conventional direct Macintosh laryngoscope (direct laryngoscopy) in patients with a predicted difficult airway. METHODS: One hundred and twenty adult patients undergoing elective minor surgery requiring general anaesthesia and endotracheal intubation presenting with at least one predictor for a difficult airway were enrolled after Institutional Review Board approval and written informed consent was obtained. Repeated laryngoscopy was performed using direct laryngoscope, DCI laryngoscope and GlideScope in a randomized sequence before patients were intubated. RESULTS: Both video laryngoscopes showed significantly better laryngoscopic view (according to Cormack and Lehane classification as modified by Yentis and Lee = C&L) than direct laryngoscope. Laryngoscopic view C&L >or= III was measured in 30% of patients when using direct laryngoscopy, and in only 11% when using the DCI laryngoscope (P < 0.001). The GlideScope enabled significantly better laryngoscopic view (C&L >or= III: 1.6%) than both direct (P < 0.001) and DCI laryngoscopes (P < 0.05). Clinically relevant improvement in the specific 36 patients with insufficient direct view (C&L >or= III) could be achieved significantly more often with the GlideScope (94.4%) than with the DCI laryngoscope (63.8%; P < 0.01). Laryngoscopy time did not differ between instruments [median (range): direct laryngoscope, 13 (5-33) s; DCI laryngoscope, 14 (6-40) s; GlideScope, 13 (5-34) s]. In contrast, tracheal intubation needed significantly more time with both video laryngoscopes [DCI laryngoscope, 27 (17 94) s, P < 0.05 and GlideScope, 33 (18-68) s, P < 0.01] than with the direct laryngoscope [22.5 (12-49) s]. Intubation failed in four cases (10%) using the direct laryngoscope and in one case (2.5%) each using the DCI laryngoscope and the GlideScope. CONCLUSION: We conclude that the video laryngoscope and GlideScope in particular may be useful instruments in the management of the predicted difficult airway. PMID- 19809329 TI - Intramuscular injection of malignant hyperthermia trigger agents induces hypermetabolism in susceptible and nonsusceptible individuals. AB - BACKGROUND AND OBJECTIVE: A new minimally invasive metabolic test for the diagnosis of susceptibility for malignant hyperthermia measuring intramuscular p(CO(2)) and lactate following local application of caffeine and halothane in humans was recently proposed. The present study tested the hypothesis that a more simplified test protocol allows a differentiation between malignant hyperthermia susceptible (MHS) and malignant hyperthermia nonsusceptible (MHN) and control individuals. METHODS: With approval of the local ethics committee and informed consent, microdialysis and p(CO(2)) probes with attached microtubing were placed into the lateral vastus muscle of six MHS, seven MHN and seven control individuals. Following equilibration, boluses of 500 microl caffeine 80 mmol l( 1) and halothane 10 vol% dissolved in soybean oil were injected locally. p(CO(2)) and lactate were measured spectrophotometrically. RESULTS: The maximal rate of p(CO(2)) increase was significantly higher in MHS than in MHN and control individuals following application of halothane and caffeine, respectively. Intramuscular caffeine injection leads to a significantly higher increase of local lactate levels in MHS than in MHN and control individuals, whereas halothane increased local lactate levels in all investigated groups. Haemodynamic and systemic metabolic parameters did not differ between the investigated groups. CONCLUSION: Local caffeine and halothane injection increased intramuscular metabolism in MHS individuals significantly more than in the two other groups. In contrast to previous investigations, direct injection of the concentrations of halothane described here increased lactate and p(CO(2)) even in MHN skeletal muscle. PMID- 19809330 TI - Body mass index and waist circumference predict both 10-year nonfatal and fatal cardiovascular disease risk: study conducted in 20,000 Dutch men and women aged 20-65 years. AB - AIMS: Body mass index (BMI) and waist circumference (WC) are both predictors of cardiovascular diseases (CVD). We compared absolute risk, hazard ratio (HR), and population attributable risk of nonfatal and fatal CVD for BMI and WC in a large prospective cohort study with an average follow-up of 10 years. METHODS AND RESULTS: Anthropometric data were measured between 1993 and 1997 in a general population sample of over 20,000 men and women aged 20-65 years in the Netherlands. All risks were adjusted for age and sex. Absolute risk of nonfatal CVD was on average 10 times higher than that of fatal CVD. In obese respondents (BMI >or=30 kg/m2), relative risk of fatal CVD was four-fold higher [HR=4.0 95% confidence interval (CI)=2.4-6.6], whereas risk of nonfatal CVD was two-fold higher (HR=1.8 95% CI=1.6-2.2) than in normal-weight respondents. Similar associations were observed for WC (>or=88 vs. <80 cm in women and >or=102 vs. 94 cm in men). In persons with overweight or obesity (BMI >or=25 kg/m2), half of all fatal CVD (attributable risk=54%, 95% CI=30-70) and a quarter of nonfatal CVD was ascribed to their overweight. On the population level, one-third of all fatal CVD cases could be attributed to overweight and obesity (population attributable risk=35%, 95% CI=14-52), and about one in seven of nonfatal CVD cases. CONCLUSION: The associations of BMI and WC with CVD risk were equally strong. Overweight and obesity had a stronger impact on fatal CVD than on nonfatal CVD. PMID- 19809331 TI - Independent associations of physical activity and cardiovascular fitness with cardiovascular risk in adults. AB - BACKGROUND: Uncertainty still exists whether physical activity (PA) and cardiovascular fitness (CF) contribute separately to cardiovascular disease (CVD) risk. This study examined the associations of PA and CF on individual as well as clustered CVD risk factors. DESIGN: Cross-sectional. METHODS: Seven hundred and eighty-one men and 890 women, aged 20-65 years, from two random population-based samples of Swedish women and men were included. PA was assessed by questionnaire and CF was predicted by a submaximal cycle ergometry test. Waist circumference, blood pressure, and fasting levels of blood lipids were assessed and dichotomized by conventional cut-off points. RESULTS: Participants reporting high PA level benefited from lower triglycerides and atherogenic cholesterol levels, regardless of CF. Higher CF level was, regardless of PA, associated with lower risk for all risk factors. With regard to clustering of risk factors, each higher CF level was associated with a gradually reduced risk by half or more, independent of PA. Furthermore, being unfit but reporting high PA was associated with a 50% lower risk compared with being unfit and inactive. Furthermore, high reported PA was associated with an additional reduced risk among fit participants. In addition, an excess risk of interaction was found for waist circumference, triglycerides, and the clustered CVD risk between neither being sufficiently active nor being fit. CONCLUSION: This study suggests that both PA and CF are independently associated with lower cardiovascular risk, and that both variables should be taken into account when CVD risk is estimated. PMID- 19809332 TI - Pathological and physiological hypertrophies are regulated by distinct gene programs. AB - BACKGROUND: This study aims to investigate changes that occur during progression and establishment of physiological and pathological cardiac hypertrophy, by microarray technology and functional annotations. DESIGN AND METHODS: Myocardial infarction leading to heart failure was induced in rats, with animals killed 1, 3, 7, 14, 42, and 92 days after coronary artery ligation. A second group was subjected to daily treadmill exercise and killed 1, 4, 24, and 48 h after a single exercise bout, or after 28 or 56 days of exercise training. RESULTS: Physiological hypertrophy was associated with less transcriptional alternation than pathological hypertrophy, indicating that posttranscriptional and translational regulation may be more important. The main difference between the two types of hypertrophy was that myocardial infarction was associated with downregulation of genes related to fatty acid metabolism, whereas no such change occurred after exercise training. Thus, fatty acid metabolism may distinguish adverse maladaptive hypertrophy from beneficial adaptive hypertrophy. CONCLUSION: This study points to specific genes and gene classes related to biological processes that may be important in these well-characterized rat models of physiological and pathological cardiac hypertrophy. PMID- 19809333 TI - Remote mobile health service utilization post 2005 Kashmir-Pakistan earthquake. AB - AIMS: Limited published studies have examined patient characteristics and disease patterns served by mobile clinics in remote areas post natural disaster. By comparing users of a remote mountainous stationary clinic and a mobile helicopter outreach clinic in Neelam Valley, 4 months post Pakistan-Kashmir earthquake, this study aims to: (i) compare the demographic and disease profile of health service users, (ii) examine how acute and chronic disease patterns vary with clinical settings, and (iii) discuss the potential implications to mobile emergency clinical service planning acute phase of natural disaster in remote areas. METHODS: Cross-sectional, retrospective record-based study in two remote clinical settings in Neelam Valley, Kashmir-Pakistan. Patients who presented to clinics from 21 January to 20 February, and whose diagnoses were available, were included in the study. Descriptive statistics and chi significance tests were conducted. RESULTS: Different users and disease profiles were found between mobile and stationary clinics. While older age, female sex, living further way from clinics, earthquake-related trauma, wound and gastrointestinal infections were found to be more significantly associated with mobile clinic users, stationary clinics patients tended to be younger, living closer to the clinic site, and disease profiles were predominantly chronic in nature. CONCLUSION: In addition to the provision of acute and chronic medical clinical service, mobile clinics operating during post acute emergency phase of natural disaster in remote areas should be sex sensitive, with highly trained and experienced staff as well as the capacity to provide trauma care. PMID- 19809334 TI - Achalasia mistaken as eating disorders: report of two children and review of the literature. AB - Eating disorders are commonly considered diagnoses in young women who present with unexplained weight loss and vomiting. Our objective was to report the increased awareness of eating disorders and that it is likewise important to recognize that organic pathology (achalasia) can cause symptoms that may mimic an eating disorder and lead to misdiagnosis. Two case reports are presented and a review of the existing literature is provided. In the first patient, initial diagnosis of nonclassified eating disorder based on a pubertal conflict was made, and 3.5 years later diagnosis of primary achalasia was established. Atypical bulimia nervosa was initially suspected in the other case, but diagnosis of achalasia was established at an early stage of evaluation. The exclusion of organic disease must be a priority, even if a psychotherapeutic intervention may be needed in the global care of eating disorder patients. Esophageal achalasia should be considered in anyone presenting with difficulty swallowing or dysphagia, even if other features suggest anorexia nervosa or bulimia nervosa. PMID- 19809335 TI - Toll-like receptor 4 gene polymorphism modulates phenotypic expression in patients with hereditary hemochromatosis. AB - BACKGROUND: Clinical penetrance of hereditary hemochromatosis is highly variable. We hypothesized that it might be modified by factors involved in the cellular immune response, such as toll-like receptors (TLRs) or nucleotide oligomerization domain proteins (NODs). METHODS: Clinical expression of hemochromatosis was assessed as a function of TLR4, TLR9, and NOD2 polymorphisms in 99 homozygous carriers of the HFE C282Y mutation with mild-to-severe iron overload. RESULTS: Thirteen (13%) of the 99 hemochromatosis patients were heterozygous for a TLR4 Asp299Gly polymorphism and 86 (87%) were TLR4 wild-type-only carriers. Clinical expression of hemochromatosis was observed more frequently in carriers of the TLR4 polymorphism (100%) than in TLR4 wild-type carriers (56%, P = 0.002). This was based on higher prevalences of liver disease (92 vs. 45%, P = 0.002) and arthropathy of metacarpophalangeal joints (69 vs. 35%, P = 0.018) in TLR4 polymorphism carriers. The finding was strengthened by the strong association of TLR4 polymorphism with liver fibrosis in the subgroup of 52 patients who underwent a liver biopsy (P = 0.011). The TLR4 polymorphism did, however, not correlate with body iron overload. The study results remained significant in multiple regression analyses after excluding possible confounding effects, such as age, sex, alcohol, or meat intake, and in the subgroup of 84 patients presenting as the first members of their families. CONCLUSION: TLR4 Asp299Gly polymorphism modulates clinical expression in patients with hereditary hemochromatosis. The polymorphism does not correlate with iron overload suggesting that TLR4 plays a role in an inflammatory process arising from toxic effects of iron accumulation. PMID- 19809336 TI - Psychosocial-spiritual factors in patients with functional dyspepsia: a comparative study with normal individuals having the same endoscopic features. AB - OBJECTIVES: The aim of the study is to identify factors that are associated with dyspeptic symptoms in patients with functional dyspepsia (FD) in a multivariate context. METHODS: Demographic data, personal habits, stressful life events, and psychological distress were compared between functional dyspeptic patients with gastric reddish streaks and asymptomatic counterparts who underwent upper gastrointestinal endoscopy as part of a self-paid physical check-up. RESULTS: There were 93 patients in the symptomatic group and 67 patients in the asymptomatic group. FD patients had a lower proportion of tea consumption (38 vs. 61%, P = 0.004), more were single (20 vs. 6%, P<0.05), less belief in religion (46 vs. 66%, P<0.05), a greater number (median+/-interquartile range, 3.0+/-2.0 vs. 2.0+/-2.0, P<0.001) and more severity (1.5+/-0.9 vs. 1.0+/-1.0, P<0.001) with regard to stressful life events, greater scores of symptom dimensions of somatization, depression, anxiety, and psychotism and general severity index of psychopathology as compared with asymptomatic counterparts. Only tea consumption [odds ratio (OR) = 0.33, 95% confidence interval (CI) = 0.15-0.72, P<0.01)], religion (OR = 0.42, 95% CI = 0.19-0.91, P<0.05), number of stressful life events (OR = 2.74, 95% CI = 1.28-5.88, P<0.01), and somatization (OR = 6.80, 95% CI = 1.21-38.08, P<0.05) remained statistically significant in multivariate analysis. CONCLUSION: FD with gastric reddish streaks exhibited increased somatization, more stressful life events, less belief in religion, and less tea consumption as compared with asymptomatic counterparts. The findings of the study suggest the importance of adopting a more comprehensive holistic bio-psycho-socio-spiritual model when dealing with FD patients. PMID- 19809337 TI - A trial of quetiapine compared with risperidone in the treatment of first onset psychosis among 15- to 18-year-old adolescents. AB - The objective of this study was to evaluate the efficacy and tolerability of quetiapine in the treatment of first onset psychosis in older adolescents using risperidone as a comparator. Twenty-two patients with first onset psychosis were randomized to receive quetiapine (up to 800 mg/day) or risperidone (up to 6 mg/day) for 6 weeks. Raters blind to treatment assignment performed outcome symptom ratings. No statistical differences emerged in terms of efficacy or tolerability between the two drugs. However, there were some clinically notable differences that seem to favour the efficacy of risperidone over quetiapine. Patients taking quetiapine, although improved, showed less clinical improvement on scores for total positive and negative symptoms, clinical global severity and depression at 6 weeks than patients taking risperidone. Although both treatments were associated with weight gain and sedation, more patients on quetiapine experienced over 10% weight gain. However, fewer patients who were taking quetiapine required anticholinergic medication or experienced extrapyramidal side effects than patients taking risperidone. Risperidone was significantly more likely to be associated with elevation in serum prolactin levels in this population. In conclusion, the results in this small trial show that adolescent patients may benefit more from treatment with risperidone than quetiapine. However, those susceptible to side effects, particularly hyperprolactinaemia, may be more suitable for treatment with quetiapine. PMID- 19809338 TI - The redevelopment and validation of the Rehabilitation Adherence Questionnaire for injured athletes. AB - This study was designed to redevelop and examine the validity of the Rehabilitation Adherence Questionnaire (RAQ), which can be used in both research and medical rehabilitation settings. The participants in the study were composed of 240 injured athletes (first analysis 120, second analysis 120 athletes) who participated in a physical rehabilitation program. Scale development, descriptive analysis, and factor analysis (exploratory factor analysis, confirmatory factor analysis) developed 25 items of RAQ for injured athletes. RAQ had six subscales such as support from significant others (five items), pain tolerance (five items), scheduling (four items), self-motivation (five Items), perceived exertion (three items), and environmental conditions (three items). Criterion-related validity, differential validity, and test-retest reliability of RAQ indicated that RAQ can be used both in clinic-based study and rehabilitation settings. In summary, this study showed good psychometric properties of RAQ. PMID- 19809339 TI - High-pitch electrocardiogram-triggered computed tomography of the chest: initial results. AB - OBJECTIVES: Chest pain is one of the most frequent symptoms in the emergency department. A variety of different diseases, some of them acutely life threatening, can be the underlying cause. Electrocardiogram (ECG)-gated computed tomography angiography of the thorax has been proposed as a cost and time effective imaging technique for these patients. We describe a new high-pitch scan mode, which has been developed specifically for low-dose ECG-triggered computed tomography angiography using dual source computed tomography (CT). MATERIAL AND METHODS: Twenty-four patients were examined with this technique on a second generation dual source CT system. The scan mode uses a pitch of 3.2 to acquire a spiral CT data set of the complete thorax in less than 1 second with a temporal resolution of 75 ms (scan parameters: 128 x 0.6 mm collimation, 0.28 seconds gantry rotation time, 370 mAs at 100 kV [15 patients] and 320 mAs at 120 kV [9 patients], reconstructed slice thickness 0.6 mm, increment 0.4 mm). Data acquisition was prospectively triggered at 50% to 60% of the RR interval to cover the range over the heart in diastole. A triple phase contrast injection protocol (total volume: 80 mL) was used to optimize enhancement of the pulmonary and systemic arterial vessels. Image quality was evaluated using a 4-point scale (1 = absence of motion artifacts; 2 = slight motion artifacts, fully evaluable; 3 = motion artifacts, but evaluable; 4 = unevaluable) on a per-segment basis. RESULTS: The patients had an average heart rate of 68 +/- 15 bpm (range: 43-111 bpm) during data acquisition. Motion artifact free visualization of the aorta and pulmonary vessels was possible in each case, of 344 coronary artery segments, 242 (70%) had an image quality score of 1, 60 segments (17%) a score of 2, 28 segments (8%) a score of 3, and 14 segments (4%) were rated as "unevaluable." In 17 patients (10 patients with a heart rate < or =60 bpm) all segments were evaluable. The average dose length product was 113 +/- 11 mGy x cm per scan (mean effective dose 1.6 +/- 0.2 mSv) at 100 kV and 229 +/- 31 mGy x cm per scan (mean effective dose 3.2 +/- 0.4 mSv) at 120 kV. CONCLUSION: Our initial results indicate that this high-pitch scan mode allows motion artifact free and accurate visualization of the thoracic vessels, and diagnostic image quality of the coronary arteries in patients with low and stable heart rates at a very low radiation exposure. PMID- 19809340 TI - Comprehensive assessment of renal function and vessel morphology in potential living kidney donors: an MRI-based approach. AB - OBJECTIVES: To test the feasibility of a single MR-based examination allowing for the comprehensive assessment of renal anatomy, function, and vascular status in candidates for kidney transplantation. MATERIALS AND METHODS: Twelve healthy potential kidney donors (mean age: 47 +/- 14 [standard deviation] years, 3 men and 9 women) participated in the study, which was approved by the local ethics committee. MR-nephrography was performed using a navigator-gated T1-weighted saturation-recovery sequence (TrueFISP: TR/TE = 621.31 milliseconds/1.27 milliseconds, Flip angle = 70 degrees, TI = 300 milliseconds, BW = 977 Hz/Px, or TurboFLASH: TR/TE = 528 milliseconds/1.15 milliseconds, Flip angle = 8 degrees, TI = 300 milliseconds, BW = 600 Hz/Px). Images were acquired up to 60 minutes after the injection of 4 mL of gadobutrol. The glomerular filtration rate (GFR) was evaluated from the renal clearance of gadobutrol within the extra cellular fluid volume by exponential fitting of time-signal curves measured over the liver. MR-angiography was performed using a T1-weighted 3D-Flash sequence. The overall measuring time was 70 to 80 minutes. For each subject, GFR data were compared with the results of renal scintigraphy with Tc-labeled DTPA from the same day. RESULTS: Kidney anatomy and vascular status were successfully assessed in all subjects. The results of MR-angiography were in excellent accordance with the surgical findings. MR-nephrography presented a good agreement to the scintigraphy (mean GFR from MR-nephrography = 117 +/- 24 mL/min per 1.73 m; mean GFR from scintigraphy = 116 +/- 26 mL/min per 1.73 m). The Bland-Altman-plot showed a mean difference in measurements pairs of -1 +/- 12 mL/min per 1.73 m. The absolute paired difference ranged between 0 and 22 mL/min per 1.73 m. CONCLUSIONS: The study showed the feasibility of the comprehensive assessment of renal anatomy, function, and vascular morphology, using 1 single MR examination. The proposed protocol may find immediate clinical application in the preoperative assessment of potential kidney donors. PMID- 19809341 TI - Dual source CT coronary angiography in severely obese patients: trading off temporal resolution and image noise. AB - OBJECTIVE: To assess in severely obese patients the subjective and objective image quality parameters and to estimate the radiation dose of dual-source computed tomography coronary angiography (CTCA), using 3 different protocols. MATERIALS AND METHODS: Dual-source CTCA was performed in 60 patients (30 women; mean age 58 +/- 7 years) suffering from obesity class II or higher (body mass index [BMI] >35 kg/sq m). Twenty patients were examined with a standard CTCA protocol at 120 kV/350 mAs (protocol A), 20 patients with a CTCA protocol at 140 kV/350 mAs (protocol B), and 20 patients at 140 kV/350 mAs with a dedicated obesity protocol (protocol C), that allows the additional data sampling by expanding the data acquisition for each tube from a quarter to a half rotation, permitting to trade off temporal resolution and image noise. Two blinded observers independently assessed the image quality of each coronary segment, using a 4-point scale (1: excellent-4: nondiagnostic) and measured the different image parameters (image noise, signal-to-noise ratio [SNR], and contrast-to-noise ratio [CNR]). Radiation dose estimates were calculated. RESULTS: The average BMI was 46.3 +/- 8.3 kg/sq m (range, 36.8-69.6 kg/sq m). Subjective image quality (1.55 +/- 0.73) was significantly better in protocol C when compared with protocol A (2.46 +/- 0.76; P < 0.01) and protocol B (2.12 +/- 0.87; P < 0.017). There was a significantly lower rate of coronary artery segments with nondiagnostic image quality when using the obesity protocol C (1.5%; 4/262) compared with that obtained when using protocol A (7.8%; 22/280; P < 0.01) and protocol B (4.4%; 12/275; P < 0.017). Image noise was significantly lower in protocol C (31.8 +/- 5.0 HU) when compared with group A (43.5 +/- 4.7 HU; P < 0.001) and B (36.8 +/- 5.5 HU; P < 0.01). SNR and CNR were significantly higher in group C (13.8 +/- 2.4 and 23.1 +/- 2.8) compared with group A (10.6 +/- 1.7 and 15.1 +/- 3.2; each P < 0.001) and group B (12.0 +/- 2.0 and 18.8 +/- 3.1; each P < 0.01). The estimated effective radiation dose of the obesity protocol C (15.6 +/- 0.9 mSv) was significantly higher when compared with that in protocol A (10.1 +/- 0.8 mSv; P < 0.01), but not significantly different from that in protocol B (13.3 +/- 0.8 mSv; P = 0.022). CONCLUSIONS: Use of an obesity protocol in dual-source CTCA in severely obese patients significantly improves image quality, but goes along with a higher radiation dose. PMID- 19809342 TI - An eight-channel phased array RF coil for spine MR imaging at 7 T. AB - OBJECTIVE: To develop a transmit/receive radiofrequency (RF) array for MR imaging of the human spine at 7 T. The prototype is characterized in simulations and bench measurements, and the feasibility of high-resolution spinal cord imaging at 7 T is demonstrated in in vivo images of volunteers. MATERIALS AND METHODS: The RF phased array consists of 8 overlapping surface loop coils with a dimension of 12 cm x 12 cm each. Bench measurements were obtained with a phantom made of body simulating liquid and assessed with a network analyzer. For safety validation, numerical computations of the RF field distribution and the corresponding specific absorption rate were performed on the basis of 3 different human body models. In vivo images of 3 volunteers (2 with a documented scoliosis) were acquired using a 3D-FLASH sequence with a high spatial resolution of 0.57 mm isotropic. RESULTS: The 7 T transmit/receive RF coil could be easily integrated into the patient table for examinations of the cervicothoracic or thoracolumbosacral spine. Comparable results were found for all 3 numerical calculations using different human body models. Measurements of the g-factor indicated good image quality for parallel imaging acceleration factors up to 2.7 along the head-feet direction, which could be validated in the in vivo images. The in vivo images demonstrated very fine anatomic features such as the longitudinal ligaments or the venous drainage through the vertebral bodies. A largely homogeneous excitation over an extensive field-of-view of 40 cm could be obtained. CONCLUSIONS: These early results indicate that a multichannel transmit/receive phased array RF coil can be used for in vivo spine imaging at 7 T, thereby rendering high-resolution spine imaging a promising new application in 7 T clinical research. PMID- 19809343 TI - Simultaneous dynamic blood oxygen level-dependent magnetic resonance imaging of foot and calf muscles: aging effects at ischemia and postocclusive hyperemia in healthy volunteers. AB - PURPOSE: To demonstrate the feasibility of simultaneous blood oxygen level dependent (BOLD) magnetic resonance imaging of calf and foot muscles and investigate age-related changes of BOLD signal changes during ischemia and postocclusive hyperemia in healthy volunteers. MATERIAL AND METHODS: In this study, 15 healthy elderly volunteers (mean age: 69.0 +/- 7.4 years) and 15 healthy young volunteers (mean age: 26.1 +/- 3.9 years) were enrolled. In both legs, simultaneous BOLD imaging of calf and foot muscles was performed at 1.5 Tesla. Short-term ischemia and consecutive reactive hyperemia were provoked by a cuff-compression paradigm. T2*-weighted signal time courses were obtained from foot and calf muscles simultaneously. Ischemia was assessed by T2* minimum ischemic value (MIV) and the time to half ischemic minimum (THIM). Reperfusion was assessed by the time to reach T2* half hyperemia peak (THHP). Reactive hyperemia was characterized by hyperemia peak value (HPV), time to peak (TTP), and relative T2* change from end of ischemia to HPV (deltaS). Parameter differences were assessed using a 2-sided Student t test. RESULTS: Dynamic BOLD measurement of foot and calf muscles was techniqually feasible and successful in all volunteers.In comparison, THIM was significantly longer in elderly than in young volunteers for calf (P < 0.01) muscles (young: 28.9 +/- 3.7 seconds; elderly: 57.8 +/- 31.4 seconds) and foot (P = 0.01) muscles (young: 36.8 +/- 25.5 seconds; elderly: 56.6 +/- 31.7 seconds). MIV relative to baseline T2*-signal was significantly (P < 0.01) lower in the elderly for calf (young: 96.0% +/- 2.6%; elderly: 91.3% +/- 4.4%) and foot (young: 95.8% +/- 2.5%; elderly: 91.1% +/- 8.2%) muscles. TTP was significantly (calf: P = 0.01; foot: P = 0.02) delayed in the elderly (elderly calf: 103.0 +/- 92.7 seconds and foot: 157.1 +/- 109.9 seconds vs. young calf: 54.8 +/- 42.1 seconds and foot: 95.1 +/- 77.6 seconds). HPV was significantly (calf: P < 0.01 and foot: P = 0.03) higher in (young calf: 114.1% +/- 7.2% and foot: 105.8% +/- 3.3% vs. elderly calf: 104.0% +/- 2.1% and foot: 103.9% +/- 3.2% seconds) young volunteers.In a muscle-group comparison, no significant differences in THIM and MIV were observed between calf and foot.THHP was significantly (P = 0.02) longer in foot muscles (foot young: 32.5 +/- 29.8 seconds and elderly: 34.1 +/- 25.0 seconds vs. calf young: 16.8 +/- 14.1 seconds and elderly: 23.6 +/- 14.1 seconds) of both age groups. TTP was significantly (P = 0.01 and 0.02) longer in foot muscles (foot young: 95.1 +/- 77.6 seconds and elderly: 157.1 +/- 109.9 seconds vs. calf young: 54.8 +/- 42.1 seconds and elderly: 103.0 +/- 92.7 seconds) of both age groups. HPV was lower (P < 0.01) in foot muscles of the young (calf: 114.1% +/- 7.2% vs. foot: 105.8 +/- 3.3%). CONCLUSION: Simultaneous BOLD-imaging of calf and foot muscles is feasible and reveals statistically significant age-related differences during ischemia and postocclusive hyperemia in healthy volunteers. PMID- 19809344 TI - Thrombus imaging with fibrin-specific gadolinium-based MR contrast agent EP 2104R: results of a phase II clinical study of feasibility. AB - PURPOSE: To determine the feasibility of detecting thrombi using a fibrin specific gadolinium-based magnetic resonance imaging contrast agent, EP-2104R. METHODS: Subjects with confirmed thrombus in the venous system (n = 14), or in the heart, or arterial system (n = 38) were enrolled. Patients were imaged before and at various times following a 4 mumol/kg intravenous bolus injection of EP 2104R: <1 hour (N = 16), 2 to 6 hours (N = 36), and/or 20 to 36 hours (N = 33). Images were assessed by investigators at each site and by a single reader not affiliated with the sites to determine whether thrombi were visible, not visible, or further enhanced with EP-2104R. A subset of data was analyzed quantitatively by measuring a signal intensity relative to background tissue. RESULTS: Overall, 29 thrombi were visible before contrast administration, 3 of 14 in the venous system, and 26 of 38 in the arteries and heart. Thrombi generally enhanced in signal after EP-2104R injection, and an additional 7 were visualized. After contrast, 4 of 14 thrombi were visible in the venous system, and 32 of 38 in the arteries and heart. Thrombi were more conspicuous when imaged at 2 to 6 hours post EP-2104R compared with within 1 hour, because of lower blood background. Quantitatively, the post: pre signal intensity ratio was 1.90 at 2 to 6 hours post injection (standard deviation = 1.08, N = 20, P < 0.001); and 2.04 (standard deviation = 1.29, N = 19, P < 0.0025) for the 20 to 36 hours time point. There were no serious adverse events considered related to study drug. CONCLUSION: EP 2104R enhanced magnetic resonance imaging detects thrombi not readily visible in precontrast screening and gives additional enhancement of thrombi that are visible in precontrast imaging. PMID- 19809345 TI - Metal artifact reduction from reformatted projections for hip prostheses in multislice helical computed tomography: techniques and initial clinical results. AB - PURPOSE: Hip prosthesis is one of the most common types of metal implants and can cause significant artifacts in computed tomography (CT) examinations. The purpose of this work was to develop a projection-based method for reducing metal artifacts caused by hip prostheses in multislice helical CT. METHOD AND MATERIALS: The proposed method is based on a novel concept, reformatted projection, which is formed by combining the projection data at the same view angle over the full longitudinal scan range. Detection and segmentation of the metal were performed on each reformatted projection image. Two dimensional interpolation based on Delaunay triangulation was used to fill voids left after removal of the metal in the reformatted projection. The corrected data were then reconstructed using a commercially available algorithm. The main advantage of this method is that both the detection of the metal objects and the interpolations are performed on complete reformatted projections with the entire metal region present, which is particularly useful for long hip prostheses. Twenty clinical abdominal/pelvis exams with hip prostheses were corrected and clinically evaluated. RESULTS: The overall image quality and the conspicuity in some critical organs were significantly improved compared with the uncorrected images: overall quality (P = 0.0024); bladder base (P = 0.0027), and rectum (P = 0.0078). The average noise level in the bladder base was reduced from 86.7 HU to 36.2 HU. In 17 of 20 cases, the radiologists preferred either coronal (13) or axial (4) views of the corrected images. CONCLUSIONS: A novel method for reducing metal artifact in multislice helical CT was developed. Initial clinical results showed that the proposed method can effectively reduce the artifacts caused by metal implants for the cases of unilateral and bilateral hip prothesis. PMID- 19809346 TI - Correlation of fat distribution in whole body MRI with generally used anthropometric data. AB - OBJECTIVES: : Obesity is a commonly known risk for many diseases such as metabolic syndrome and cardiovascular disease. Especially important is the discrimination of the adipose tissue inside the abdomen and the subcutaneous adipose tissue. Aim of this study was to compare the whole body fat distribution, and the volume of different adipose tissue compartments respectively, with anthropometric data. MATERIALS AND METHODS: : Sixty-eight volunteers (20 males, 48 females, 42.3 +/- 15.4 years) were investigated in the context of 2 whole body magnetic resonance imaging (MRI) studies which compared the body fat distribution of depressive and bulimic patients with healthy controls. Unpublished data acquired in these studies were analyzed retrospectively.The sample consisted of 38 healthy volunteers, 17 patients with a depressive syndrome and 13 women suffering from bulimia nervosa. Individual body volume, total adipose tissue (TAT) volume, subcutaneous adipose tissue (SCAT) volume at the trunk, and visceral adipose tissue (VAT) volume were determined, using whole body MRI. Additionally, body fat profiles were standardized and a mean body distribution was calculated. Other modalities to acquire body fat content were: skin fold caliper, body impedance (3 different devices) and simple anthropometric data (Waist to Hip Ratio [WHR], Body Mass Index [BMI], distance of the aponeurosis of the rectus abdominis muscle to the ventral rim of the abdominal aorta (measured in MRI images on umbilical level) (AD) and subcutaneous adipose tissue thickness at the same level). The different modalities were correlated with the MRI data. RESULTS: : There were highly significant correlations between the skin fold data and TAT (Spearman coefficient 0.668, P >/= 0.0004) and SCAT (0.662, P >/= 0.0004). But there was no correlation with VAT. Impedance data revealed significant correlations of TAT and SCAT (Spearman 0.7, P >/= 0.0004).Simple anthropometric data like waist and hip circumference, WHR, and BMI revealed significant correlations (Spearman coefficient around 0.7-0.4, P < 0.05) with the fat compartments TAT, VAT, and SCAT.The standardized body fat slices and the VAT slices were correlated with the anthropometric data and impedance data to explore specific areas along the body axis where the correlations were higher or weaker. Skinfold data, BMI, and body impedance data yielded significant correlations with TAT along the whole body axis, as well as with VAT in almost the whole analyzed area. However, there was no special body region with locally higher correlations. WHR depicted high correlations with whole VAT, and regional TAT at the abdomen (and not with the other body regions) especially in women. Therefore, it seems to be the best marker for abdominal fat and VAT in this study. CONCLUSIONS: : We compared different body measures and body fat devices with the whole body fat distribution acquired by MRI. Generally, there were significant correlations of all modalities with body fat content (TAT) and mainly with SCAT. Correlations with VAT compartment were much weaker and an adequate estimation of VAT is, therefore, not possible. Only WHR revealed significant correlations with the fat in the body center, but only in women. If it is important to investigate especially the VAT which is responsible for a higher cardiovascular risk, risk for a metabolic syndrome and that is correlated with the course of different psychiatric diseases, cross sectional techniques such as MRI can not be substituted by simpler methods. PMID- 19809347 TI - Inspiratory muscle training improves oxygen uptake efficiency slope in patients with chronic heart failure. AB - PURPOSE: Inspiratory muscle training (IMT) improves exercise capacity and ventilatory responses to exercise in patients with chronic heart failure (CHF) with inspiratory muscle weakness (IMW). We analyzed the effects of IMT on the oxygen uptake efficiency slope (OUES) in this patient population. METHODS: Thirty two CHF patients with IMW (maximal inspiratory pressure [PImax] < 70% of predicted) were randomly assigned to either a 12-week program of IMT (IMT, n = 16) or placebo-IMT (P-IMT, n = 16). PImax and OUES were obtained before and after the intervention. RESULTS: Inspiratory muscle training resulted in 115% increment in PImax (5.9 +/- 0.9 vs 12.7 +/- 0.9 kPa; P < .001) and in significant improvement in OUES (1,554 +/- 617 to 2,037 +/- 747 mL min O2/L min of minute ventilation; P = .001). There were no significant changes in the P-IMT group. There was a significant association between the changes in PImax and OUES (r = 0.82, P < .01). CONCLUSION: In CHF patients with IMW, IMT results in a significant increase in OUES. PMID- 19809348 TI - Achieving interprofessional practice in cardiac rehabilitation. AB - PURPOSE: Cardiac rehabilitation staff members consider interprofessional practice to be the standard for delivering effective care. However, it is not known how interprofessional teams collaborate or what they consider to be important elements of collaboration. Thus, it is important to investigate how healthcare professionals plan and communicate care, work together, and define their roles as members of the cardiac rehabilitation teams. The purpose of this report was to provide an analysis of current literature related to interprofessional practice in cardiac rehabilitation, with a particular focus on examining the terms interprofessional practice and collaboration. METHODS: For this review, published articles in peer-reviewed journals for the preceding 20-year period were included from online databases (CINAHL, MEDLINE, EBM Reviews, PubMed, and Google Scholar). Key words used in the search included "cardiac rehabilitation," "cardiac recovery," and "interprofessional and interdisciplinary practice and collaboration." Of the 67 articles reviewed, 7 met inclusion criteria specifically addressing interprofessional practice in cardiac rehabilitation. RESULTS: Analysis revealed that (1) the terms interprofessional and multidisciplinary are commonly used interchangeably in healthcare, revealing a lack of clarity regarding interprofessional practices and approaches, and (2) there are few articles that clearly describe, define, or discuss interprofessional practice or collaboration in cardiac rehabilitation settings, rendering it difficult for practitioners to adhere to published practice guidelines. CONCLUSION: It is unclear why and how professional team members practice in specific ways to form cardiac teams. Further research is required to increase an understanding of these issues and to develop possibilities for the enhancement of cardiac rehabilitation practice. PMID- 19809349 TI - Costs of cardiac rehabilitation and enhanced lifestyle modification programs. AB - PURPOSE: Inadequate payment to providers for traditional cardiac rehabilitation (CR) and lifestyle modification programs may contribute to low utilization, but little systematic evidence exists. This article estimates and compares the per patient costs and revenues for 3 types of secondary prevention programs: the Dr Dean Ornish Program for Reversing Heart Disease (Ornish), the Benson-Henry Mind/Body Medical Institute's Cardiac Wellness Program (M/BMI), and CR. METHODS: The authors developed an Excel spreadsheet template for the costs of a secondary prevention program and calibrated it to 7 programs that provided the necessary data. The calibration was based on budgets, cost accounting, statistical reports, and structured interviews (in person or by telephone). RESULTS: The 4 lifestyle programs (2 Ornish and 2 M/BMI) cost almost 4 times as much per patient as the 3 traditional CR programs (means of $7,176 and $1,828, respectively; difference P < .05). The Ornish program costs averaged more than twice those of M/BMI ($9,895 and $4,458, respectively; difference P < .10). Medicare-allowed charges (including co-payments) were $5,650 for Ornish, $4,800 for M/BMI, and about $32.50 per session or $683 overall for CR. CONCLUSIONS: Programs achieved the lowest costs per patient by carefully matching program capacity to demand. In none of the programs did net revenues cover costs. The findings suggest that 4 patients could attend a traditional CR program for the cost of 1 patient in an enhanced program. PMID- 19809350 TI - Effect of a computerized referral at hospital discharge on cardiac rehabilitation participation rates. AB - PURPOSE: We evaluated the effectiveness of a computerized cardiac rehabilitation (CR) recruiting strategy instituted during the hospital discharge process after acute myocardial infarction (MI) or coronary artery bypass graft (CABG) surgery. The computerized referral was designed to enhance an already-established nurse based recruiting strategy that included personalized, in-hospital visits. METHODS: We determined CR participation rates before and after the implementation of the computerized referral process. The computerized intervention included a personalized letter at hospital discharge and a scheduled group introductory visit at the CR center. Data sources for the study included hospital electronic medical records, the hospital discharge summary, and the CR enrollment database RESULTS: Before the study intervention, 47% (171/364) of patients participated in the CR program, whereas, in year 2, this increased to 53% (231/440) (adjusted P < .01, odds ratio of participation = 1.49, 95% confidence interval = 1.11-1.99). Participation rates increased more in patients after MI (39%-48%, P < .05) than after CABG surgery (63%-67%, P = .69). During each time period, enrollment was higher after CABG surgery than after MI. CONCLUSIONS: Despite an aggressive baseline recruiting strategy, the addition of a computerized referral process during hospital discharge planning resulted in increased CR participation rates. These results strongly support the institution of computerized referral processes to CR during the hospital discharge planning process. PMID- 19809351 TI - Determinants of long-term survival in patients hospitalized for heart failure. AB - BACKGROUND AND OBJECTIVES: Current models used to stratify patients with heart failure are complex, difficult to use, and limited by short follow-up and patient selection. Our aim was to determine predictors of long-term survival in patients hospitalized for heart failure and to develop a simple mortality risk score to estimate long-term mortality. METHODS: We prospectively followed up, during 10 years after hospitalization, 701 patients with confirmed heart failure from the HOLA (Heart failure: Observation of Local Admissions) registry. RESULTS: Mean age was 72.4 +/- 11.7 years; 45% were men. During follow-up, 465 patients died and 5 underwent heart transplantation. A total of 231 patients (33%) were alive and transplant-free at the end of follow-up (5.2 +/- 4.2 years). Median survival was 3.2 years. Multivariate analysis showed that six variables (age, previous renal disease, previous stroke, chronic obstructive pulmonary disease, left ventricular ejection fraction and aortic stenosis) were independent predictors of shorter survival time. By dichotomizing these variables, we obtained six factors with similar predictive values (hazard ratio between 1.5 and 2.0). A risk score for mortality was developed using these predictors by assigning 1 point to each and adding the total for each patient. Median survival for patients with 0, 1, 2, and 3 or more points were 6.5, 5.5, 3.3, and 1.7 years, respectively. One-year mortality rates were 15, 20, 28, and 49%, respectively. CONCLUSION: The prognosis of patients hospitalized with heart failure is highly variable. A simple risk score, based on six variables readily obtainable on admission can effectively stratify patients according to their predicted mortality. PMID- 19809352 TI - Dominant right coronary artery occlusion entailing diffuse ST-segment elevation in the precordial leads. AB - Right ventricular infarction (RVI) during inferior myocardial infarction (MI) is readily diagnosed when ST-segment elevation (STE) is recorded in lead V4R. RVI may also yield precordial STE and such an electrocardiographic (ECG) pattern may be misinterpreted as a sign of anterior MI. We present a case of inferior-right ventricular (RV) MI due to occlusion of a dominant right coronary artery manifesting STE in the inferior, all precordial and right chest leads. RV dilation due to acute ischemic insult facilitated STE in leads V1-V4 despite the dominant opponent inferior and posterolateral left ventricular injury current. This case illustrates that dilation of an infarcted RV should be considered when such an ECG pattern is encountered during inferior MI, specifically a dominant one. Awareness of the circumstances under which this ECG pattern develops facilitates avoidance of misinterpretation as a sign of anterior MI and proper management. PMID- 19809353 TI - Ablation of atrial tachycardia originating from the noncoronary sinus: case report and literature review. AB - This report describes a patient with atrial tachycardia who underwent radiofrequency catheter ablation. Two applications of radiofrequency energy were received at different sites within 1 week. Initially, right atrium mapping indicated that the earliest activation site was located at the His-bundle region (HBR). However, the first application of radiofrequency near HBR appeared to be ineffective. Moreover, a degree III atrioventricular block and inferior ST segment elevation was temporally observed during the therapy, which recovered after drug treatment. As later aortic sinus mapping showed that the activation site located at the noncoronary sinus (NCS) preceded the one at HBR, and this activation terminated at atrial tachycardia, a second application of radiofrequency was performed at NCS without any complications. In addition, a literature review of the clinical features of atrial tachycardia at the NCS is also summarized in the present report. PMID- 19809354 TI - Is there a relationship between obesity, heart rate variability and inflammatory parameters in heart failure? AB - BACKGROUND: To investigate the effect of body mass index (BMI) on heart rate variability (HRV) and inflammatory parameters in patients with heart failure. METHODS: We analyzed 55 consecutive patients (mean age, 63.5 +/- 12.8 years; male/female, 39/16) with symptomatic left ventricular systolic (ejection fraction <45%) heart failure. The participants were classified into three categories according to BMI: lean (BMI < 25 kg/m2), overweight (BMI = 25-29.9 kg/m2) and obese (BMI >or= 30 kg/m2). The cause of heart failure was mainly ischemic heart disease (75%) with mean ejection fraction 30 +/- 7%. Plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) and C-reactive protein levels were measured, and time-domain HRV indices were determined on Holter electrocardiogram. The relationship between HRV indices and laboratory, inflammatory and echocardiographic parameters was investigated with correlation analysis. RESULTS: Age, sex, clinical characteristics (hypertension, diabetes mellitus, dyslipidemia, family history, smoking) were similar between groups. BMI was inversely correlated with NT-pro BNP levels (P = 0.001). HRV indices did not differ between groups. Correlation analysis demonstrated the relationship between HRV indices and fasting blood glucose (SDNN, SDANN, SDNNI, root mean square successive differences, VTI), C-reactive protein (SDANN, SDNNI, VTI), pulmonary artery pressure (SDNN, SDANN, VTI) levels. CONCLUSION: In systolic heart failure patients a higher BMI is associated with decreased NT-proBNP levels. Although HRV indices were not different between groups, inflammatory parameters, fasting blood glucose and pulmonary artery pressure were correlated with them. PMID- 19809355 TI - Screening for esophageal varices: is esophageal capsule endoscopy ready for prime time? PMID- 19809357 TI - Hemorrhagic complications of percutaneous radiofrequency ablation for liver tumors. AB - BACKGROUND: Although radiofrequency ablation (RFA) is widely accepted as a percutaneous treatment for liver tumors; serious complications may occur resulting in 0.1% to 0.5% mortality. This study analyzed the risk factors and management of hemorrhagic complications, such as hemoperitoneum, hemothorax, and hemobilia. METHODS: We performed 4133 RFA treatments in 2154 patients with primary and metastatic liver tumors from February 1999 to December 2007. Of these, we enrolled patients with hemorrhagic complications and reviewed their medical records thoroughly. The risk factors for each hemorrhagic complication were analyzed using unconditional logistic regression. RESULTS: Hemorrhagic complications occurred in 63 out of 4133 treatments (1.5%), including hemoperitoneum in 29 (0.7%), hemothorax in 14 (0.3%), and hemobilia in 20 (0.5%). Eleven, 8, and 4 of these patients, respectively, were categorized as major complications requiring blood transfusion or drainage. Two patients died after hemoperitoneum. Logistic regression analysis revealed large tumor size [odds ratio (OR) 1.06 per 1 mm increase in diameter] and low platelet count (OR 0.88 per 10,000/microL increase) were significant risk factors for hemoperitoneum. The location of tumor nodules was a significant risk factor for hemothorax (segment 7, OR 2.31) and hemobilia (segment 1, OR 3.30). Other factors, including the number of needle insertions or the duration of ablation, were not significant. CONCLUSIONS: Although hemorrhagic complications were relatively rare with percutaneous RFA, specific treatments, such as blood transfusion and drainage, were required in some cases. Care must be taken, especially in high-risk patients. PMID- 19809358 TI - Survival analysis of patients with duodenal gastrointestinal stromal tumors. AB - GOALS: To evaluate the survival characteristics of patients with duodenal gastrointestinal stromal tumors (GISTs). BACKGROUND: GISTs represent the most common mesenchymal neoplasms. However, duodenal GISTs are relatively rare, and few studies have been performed with a focus on duodenal GISTs. STUDY: We collected the data of 41 GIST patients including 7 duodenal cases. Clinicopathologic findings and recurrence-free survival (RFS) of duodenal GIST patients were analyzed. RESULTS: The proportion of having any symptoms was 86% in duodenum, 32% in stomach, and 56% in other GISTs (P=0.034), and the most common symptoms of duodenal GISTs were melena and anemia. The 2-year RFS rates were 51.4% in duodenal GISTs, 78.4% in stomach GISTs, and 100% in other GISTs, and duodenal GISTs showed poorer RFS than nonduodenal GISTs (hazard ratio, 5.1; log rank P=0.019). Particularly, in low-risk and intermediate-risk group, the hazard ratio of recurrence was 12.3 (log-rank P=0.010). Multivariate Cox analysis showed symptom (P=0.007), mitotic index (P=0.011), and tumor location (P=0.043) were significant prognostic factors of recurrence. CONCLUSIONS: RFS of duodenal GISTs was worse than nonduodenal GISTs. PMID- 19809359 TI - Increased serum high-density lipoprotein-cholesterol concentration in celiac disease after gluten-free diet treatment correlates with body fat stores. AB - BACKGROUND: Low high-density lipoprotein-cholesterol (HDL-C) concentration correlates with increased cardiovascular risk. A great prevalence of celiac disease (CD) was reported among patients with low HDL-C concentration, and gluten free diet (GFD) treatment seems to normalize lipid profile. We evaluated blood lipids and body composition in 26 CD patients with low HDL-C level (<1.0 mmol/L) at diagnosis and after GFD. STUDY: A case-control study. METHODS: The diagnosis was based on histologic evidence of subtotal or total duodenal villous atrophy. Patients were studied before and after GFD treatment (14.2+/-1.4 mo) with biopsy proven return to normal of the duodenal mucosa. HDL-C was enzymatically assessed after precipitation of very low-density lipoprotein and low-density lipoprotein with heparin-magnesium. Apolipoprotein (Apo)-AI level was assessed by immunoturbidimetric assay; triglycerides by an enzymatic colorimetric method. Body composition was assessed by dual-energy x-ray absorptiometry. RESULTS: Body composition improved after GFD, with increasing body weight (P<0.05) essentially owing to increased fat mass (FM) (P<0.01), rather than fat-free mass (P=0.064). Total cholesterol and HDL-C were lower in untreated compared with treated patients (P<0.001 and P<0.0001). Apo-AI level increased significantly after GFD (1.20+/-0.22 vs. 1.46+/-0.17 g/L; P<0.0001). Apo-AI, sex, and FM were all significant determinants of HDL-C level; a positive correlation (R=0.68; P<0.0001) was found between increase in HDL-C level and in FM after GFD treatment. CONCLUSIONS: Restoration of lipid profile in CD patients after GFD treatment may be explained by an increase in both Apo-AI secretion by intestinal cells and body fat stores. PMID- 19809360 TI - Optimal schedule for home blood pressure monitoring based on a clinical approach. AB - OBJECTIVE: The aim of this study was to determine the optimal schedule for home blood pressure (HBP) measurement based on a clinical approach. METHODS: Four hundred and sixty-four participants underwent HBP measurement for 7 days (duplicate measurements in the morning and in the evening), ambulatory blood pressure (ABP) monitoring, and measurement of target organ damage (echocardiography and microalbuminuria). To evaluate the optimal schedule for HBP measurement, correlations of HBP with ABP and HBP with indicators of target organ damage were calculated. RESULTS: HBP decreased slightly (day 1, 129.9/85.3 mmHg; day 7, 128.6/84.8 mmHg), whereas the association between HBP and ABP or target organ damage increased with the cumulative number of measurements. The highest correlations were obtained by using the mean of all 28 measurements, although no major increase occurred after day 4. There was no change in the correlations when the measurements performed during the first day were discarded. Morning and evening HBP correlated equally well with ABP and microalbuminuria. The mean of the first measurements on each measurement occasion was 2.3/1.2 mmHg higher (P < 0.001 for both) than the mean of the second measurements, but discarding the first measurements did not result in greater correlations. The results were similar in both hypertensive and normotensive populations. CONCLUSION: Duplicate measurements on at least 4 days in the evening and in the morning are needed to reliably estimate an individual's BP level and the risk for target organ damage. Measurements performed during the first day should not be discarded, as suggested by the current European guidelines. PMID- 19809361 TI - Hypertension incidence in Turkey (HinT): a population-based study. AB - OBJECTIVE: Hypertension incidence is an important determinant of hypertension prevalence and progression. Few studies have been published on hypertension incidence in developing countries despite the high prevalence observed. The aim of this study was to investigate the incidence of hypertension in Turkey. METHODS: The study was designed as an epidemiological cohort study which included the population of the Prevalence, awareness, treatment and control of hypertension in Turkey (PatenT) Study which had 4910 volunteers. Blood pressure measurements were performed three times and a questionnaire was used to obtain data on the present status of hypertension with regards to distributions and alterations of risk factors. RESULTS: In the present study, 4008 (81.6%) participants of the PatenT Study population were contacted after 4 years. After excluding 173 dead and 67 pregnant individuals, the study cohort comprised of 3768 individuals. The overall 4-year incidence rate of hypertension was 21.4%; it reached a maximum of 43.3% in individuals over 65 years of age. Age, initial blood pressure category, and body mass index were the best predictors of the hypertension incidence rate. Multivariate logistic regression analysis revealed that age, obesity, alcohol consumption, and living in rural areas were significant predictors of hypertension. CONCLUSION: Follow-up periods scheduled considering age, initial blood pressure category, and body mass index are important for the early determination of hypertension. As there are limited data regarding hypertension incidence in developing countries, the results of data collected in this study might serve as a model. PMID- 19809362 TI - Hypertension in seven Latin American cities: the Cardiovascular Risk Factor Multiple Evaluation in Latin America (CARMELA) study. AB - BACKGROUND: Little information is available regarding hypertension, treatment, and control in urban population of Latin America. OBJECTIVE: We aimed to compare blood pressure (BP) distribution, hypertension prevalence, treatment, and control in seven Latin American cities following standard methodology. METHODS: The Cardiovascular Risk Factor Multiple Evaluation in Latin America (CARMELA) study was a cross-sectional, epidemiologic study assessing cardiovascular risk factors using stratified multistage sampling of adult populations (aged 25-64 years) in seven cities: Barquisimeto (Venezuela; n = 1848); Bogota (n = 1553); Buenos Aires (n = 1482); Lima (n = 1652); Mexico City (n = 1720); Quito (n = 1638); and Santiago (n = 1655). The prevalence of hypertension and high normal BP were determined based on 2007 European Society of Hypertension and European Society of Cardiology definitions. RESULTS: BP increased with age in men and women; pulse pressure increased mainly in the upper age group. The hypertension prevalence ranged from 9% in Quito to 29% in Buenos Aires. One-quarter to one-half of the hypertension cases were previously undiagnosed (24% in Mexico City to 47% in Lima); uncontrolled hypertension ranged from 12% (Lima) to 41% (Mexico City). High normal BP was also evident in a substantial number of each city participants (approximately 5-15%). Majority of population has other cardiovascular risk factors despite hypertension; only 9.19% of participants have no risk factors apart from hypertension. CONCLUSION: From 13.4 to 44.2% of the populations of seven major Latin American cities were hypertensive or had high normal BP values. Most hypertensive patients have additional risk factors. Public health programs need to target prevention, detection, treatment, and control of total cardiovascular risk in Latin America. PMID- 19809363 TI - Antihypertensive effects of double the maximum dose of valsartan in African American patients with type 2 diabetes mellitus and albuminuria. AB - OBJECTIVE: The blood pressure (BP)-lowering response to renin-angiotensin aldosterone system blockade in hypertensive African-Americans is typically less than in whites. To determine whether higher than conventional doses of renin angiotensin-aldosterone system blockade can improve BP reduction in African American patients. METHODS: Hypertensive patients with type 2 diabetes and albuminuria were enrolled: 110 African-Americans (BP = 150/87 mmHg, aged 57.5 +/- 11 years) and 281 non-African-Americans (BP = 151/89 mmHg, aged 57.7 +/- 11 years). All patients received valsartan 160 mg once daily in the morning for 4 weeks, following which patients were randomized to receive one of three valsartan doses: 160, 320 or 640 mg/day (2x, maximal recommended dose) for 26 weeks. If at week 6, target BP (<130/80 mmHg) was not achieved, then other add-on antihypertensives were allowed. RESULTS: The predominant BP (DeltaSBP/DeltaDBP) reduction was observed within 4 weeks and was lesser in African-Americans (7.8 +/ 15/4.5 +/- 9 mmHg) than non-African-Americans (8.9 +/- 14/6.6 +/- 1 mmHg, P < 0.05). Greater reduction in urinary albumin excretion was observed with higher doses (320 or 640 mg); however, the responses were similar between African Americans and non-African-Americans. Use of add-on antihypertensives was higher in African-American (56%) vs. non-African-American patients (36%) with a similar rate across the three valsartan doses. From week 4-26, reduction in BP was lesser (P < 0.05) for African-American than non-African-American patients at the160-mg dose but not with 320 and 640-mg doses. CONCLUSION: In African-American patients, a lower BP reduction response was observed to conventional doses of valsartan than non-African-American patients, but at 640 mg, a higher response was observed in African-American patients than in non-African-American patients. PMID- 19809364 TI - Regulation of multiple renin-angiotensin system genes by Sry. AB - BACKGROUND AND OBJECTIVE: We demonstrated that the Sry gene complex on the spontaneously hypertensive rat (SHR) Y chromosome is a candidate locus for hypertension that accounts for the SHR Y chromosome blood pressure effect. All rat strains examined to date share six Sry loci, and a seventh Sry locus (Sry3) appears to be unique to SHR male rats. Previously, we showed that Sry1 increased activity of the tyrosine hydroxylase promoter in transfected PC12 cells, and Sry1 delivered to adrenal gland of Wistar-Kyoto (WKY) rats increased blood pressure and sympathetic nervous system activity. The objective of this study was to determine whether renin-angiotensin system genes participate in Sry-mediated effects. METHOD: Sry expression vectors were co-transfected into CHO cells with luciferase reporter constructs containing promoters of angiotensinogen (Agt 1430/+22), renin (Ren -1050/-1), angiotensin-converting enzyme (ACE) (ACE 1677/+21) and ACE2 (ACE2 -1091/+83). RESULTS: Sry1, Sry2 and Sry3 differentially upregulated activity of the promoters of angiotensinogen, renin and ACE genes and downregulated ACE2 promoter activity. The largest effect was seen with Sry3, which increased activity of angiotensinogen promoter by 1.7-fold, renin promoter by 1.3-fold, ACE promoter by 2.6-fold and decreased activity of ACE2 promoter by 0.5-fold. The effect of Sry1 on promoter activity was significantly less than that of Sry3. Sry2 activated promoters at a significantly lower level than Sry1 did. The result of either an additive effect of Sry regulation of multiple genes in the renin-angiotensin system or alterations in expression of a single gene could favor increased levels of Ang II and decreased levels of Ang-(1-7). CONCLUSION: These actions of Sry could result in increased blood pressure in males and contribute to sex differences in blood pressure. PMID- 19809365 TI - Blood pressure and pulse wave velocity values in the institutionalized elderly aged 80 and over: baseline of the PARTAGE study. AB - OBJECTIVE: The aim of the longitudinal study PARTAGE (predictive values of blood pressure and arterial stiffness in institutionalized very aged population) was to determine the predictive value of blood pressure (BP) and arterial stiffness for overall mortality, major cardiovascular events and cognitive decline in a large population of institutionalized patients aged 80 and over. In the study herein, we present the baseline data values of this study. METHODS: A total of 1130 patients were recruited (878 women), living in French and Italian nursing homes. Clinical and 3-day self-measurements of BP were conducted. Aortic and upper limb pulse wave velocity were obtained using a PulsePen tonometer. RESULTS: Of this population, 76% of women and 60% of men had a known hypertension and over 91% of the patients were under antihypertensive treatment; 51% of the treated hypertensive patients were well controlled (systolic BP <140 mmHg). No significant differences were found between clinical and self-measured BP. With age, there was an increase in pulse pressure (P < 0.001) due to a decrease in diastolic BP (P < 0.001), without any increase in systolic BP. Aortic but not peripheral pulse wave velocity significantly increased with age (P < 0.005). CONCLUSION: Baseline values obtained herein demonstrate that elderly patients living in nursing homes present hemodynamic characteristics which are different to those described in community-living elderly populations, and indicate the interest of assessing, in longitudinal studies, the role of BP and arterial stiffness in morbidity and mortality in this population. PMID- 19809366 TI - Assessment of left ventricular structure and function in preeclampsia by echocardiography and cardiovascular biomarkers. AB - AIM: To assess left ventricular (LV) structure and function in preeclampsia, a serious vascular-related pregnancy disorder, by Doppler tissue imaging (DTI) in combination with the levels of cardiovascular biomarkers. MATERIAL AND METHODS: Thirty-five pregnant women with preeclampsia and 30 with normal pregnancy, matched for age and gestational age were examined during pregnancy and 3-6 months after delivery. Transthoracic echocardiography and DTI were performed and blood levels of amino-terminal pro-brain natriuretic peptide (NT-pro-BNP), C-reactive protein (CRP), cystatin C and troponin I were analyzed. RESULTS: There were significant differences in LV and left atrial dimensions and function between the groups. A higher septal and lateral E/E' ratio (E = early transmitral diastolic flow velocity and E' = early diastolic myocardial velocity) (P < 0.0001, 0.0008) and higher levels of NT-pro-BNP, cystatin C, and lower cystatin C estimated GFR in ml/min per 1.73 m(2) (P < 0.0001) were seen in the preeclampsia both during pregnancy and at follow-up. In addition the levels of E/E' ratio lateral and NT pro-BNP were higher in pregnant women with early-onset preeclampsia necessitating delivery before 34 weeks of gestation than those who developed preeclampsia and delivered at or after 34 weeks (P = 0.0004, 0.005). CONCLUSION: In pregnancies complicated by preeclampsia, especially early-onset preeclampsia, the diastolic LV function is impaired and levels of biomarkers, NT-pro-BNP and cystatin C, are increased in comparison to normal pregnancy. PMID- 19809367 TI - Insulin resistance and triglycerides. AB - In 1385 adults with primary untreated hyperlipidemia and in a population study of 339 adults (Princeton Follow-up Study [PFS]), we hypothesized that homeostasis model assessment (HOMA) insulin resistance (IR) was a significant explanatory variable for triglycerides (TG) and that IR rose in a stepwise fashion, independent of age, race, sex, and body mass index (BMI), whereas TG categories rose from less than 150 to 150 to 200, to 200 to 500, and to more than 500 mg/dL. A third hypothesis was that TG, BMI, and the ratio of TG to high-density lipoprotein cholesterol (TG/HDL-C) were significant explanatory variables for IR and that IR was inversely associated with HDL-C quintiles and positively associated with non-HDL-C quintiles. By stepwise multiple regression with age, race, sex, BMI, and IR as explanatory variables, in the 1385 patients, positive explanatory variables for TG included BMI (partial R = 1.3%, P < 0.0001), sex (men higher, partial R = 1.1%, P = 0.0001), and IR (partial R = 0.4%, P = 0.012). In the 339 PFS subjects, positive explanatory variables for TG were IR (partial R = 11.4%, P < 0.0001), race (whites higher, partial R = 2.1%, P = 0.005), and sex (men higher, partial R = 1.4%, P = 0.019). After adjusting for age, race, sex, and BMI, in 1385 patients, HOMA IR rose while TG categories rose, with least square means of 2.64 for the TG category less than 150 mg/dL, 3.27 for 150 to 200 mg/dL, 3.85 for 200 to 500 mg/dL, and 4.12 for more than 500 mg/dL. Similarly, in the PFS, while TG categories rose, the least square means of HOMA IR rose, with 1.68 for the TG category less than 150 mg/dL, 2.34 for 150 to 200 mg/dL, and 3.03 for 200 to 500 mg/dL. Body mass index, TG, and TG/HDL-C were significant explanatory variables for IR. Homeostasis model assessment IR is a significant, potentially reversible explanatory variable for TG in patients referred because of hyperlipidemia and in population subjects. PMID- 19809368 TI - Congenital abnormalities associated with hemivertebrae in relation to hemivertebrae location. AB - Numerous congenital abnormalities have been reported in association with hemivertebrae (HV). No data exist about their incidence depending on the location of the HV. From 1980 to 2003, 75 patients with 80 HV responsible for evolutive congenital scoliosis were managed by HV resection using a double approach and short anterior and posterior convex fusion. The associated abnormalities were evaluated with MRI, echocardiography and renal ultrasound. Associated genitourinary abnormalities were found in 24% of patients, cardiac abnormalities in 8% and intrathecal abnormalities in 15%. Medullar abnormalities were more frequent in case of vertebral malformations at lumbosacral level. PMID- 19809369 TI - Awareness of deficits and error processing after traumatic brain injury. AB - Severe traumatic brain injury is frequently associated with alterations in performance monitoring, including reduced awareness of physical and cognitive deficits. We examined the relationship between awareness of deficits and electrophysiological indices of performance monitoring, including the error related negativity and posterror positivity (Pe) components of the scalp-recorded event-related potential, in 16 traumatic brain injury survivors who completed a Stroop color-naming task while event-related potential measurements were recorded. Awareness of deficits was measured as the discrepancy between patient and significant-other ratings on the Frontal Systems Behavior Scale. The amplitude of the Pe, but not error-related negativity, was reliably associated with decreased awareness of deficits. Results indicate that Pe amplitude may serve as an electrophysiological indicator of awareness of abilities and deficits. PMID- 19809370 TI - Alterations in mRNA expression of BACE1, cathepsin B, and glutaminyl cyclase in mice ischemic brain. AB - The relationship between cerebral ischemia and Alzheimer's disease has been evaluated extensively. However, the association between cerebral ischemia and the deposition of beta-amyloid (Abeta) remains to be clarified. Here, we used mice bilateral common carotid artery ligation model to investigate the alterations in mRNA expression of Abeta precursor protein cleavage enzyme 1(BACE1), cathepsin B, and glutaminyl cyclase after transient global cerebral ischemia. The reverse transcriptase PCR assay showed that the expressions of these three Abeta metabolism-related genes were upregulated in brain with different manner. It indicates that all these three Abeta-metabolism-related genes may participate in the acute and chronic Abeta generation after transient cerebral ischemia, and will be helpful to understand the mechanisms underlying the linkage of brain ischemia and Alzheimer's disease. PMID- 19809371 TI - Role of Broca's area in encoding sequential human actions: a virtual lesion study. AB - The exact contribution of Broca's area to motor cognition is still controversial. Here we used repetitive transcranial magnetic stimulation (5 Hz, five pulses) to interfere transiently with the function of left BA44 in 13 healthy individuals; the task consisted of reordering human actions or nonbiological events based on three pictures presented on a computer screen and extracted from a video showing the entire sequence beforehand. We found that a virtual lesion of left BA44 impairs individual performance only for biological actions, and more specifically for object-oriented syntactic actions. Our finding provides evidence that Broca's area plays a crucial role in encoding complex human movements, a process which may be crucial for understanding and/or programming actions. PMID- 19809372 TI - Frequency tuning properties of ocular vestibular evoked myogenic potentials. AB - Loud air-conducted sound or bone-conducted vibration produces vestibular dependent electromyographic responses both in the cervical muscles (cervical vestibular evoked myogenic potentials) and in the extraocular muscles (ocular vestibular evoked myogenic potentials; oVEMPs). Although previous frequency tuning measurements of cervical vestibular evoked myogenic potentials have provided important information for a clinical application, those of oVEMPs have rarely been studied. We explored the frequency tuning properties of oVEMPs in 12 healthy participants. Our results indicate that the best frequencies of the oVEMPs to air-conducted sound and bone-conducted vibration are 500 and 250 Hz, respectively. The difference in the best frequencies between the two stimuli may originate from the difference in the end organs (saccule or utricle), preferentially activated by each stimulus. PMID- 19809373 TI - Majority of dietary glutamine is utilized in first pass in preterm infants. AB - Glutamine is a conditionally essential amino acid for very low-birth weight infants by virtue of its ability to play an important role in several key metabolic processes of immune cells and enterocytes. Although glutamine is known to be used to a great extent, the exact splanchnic metabolism in enterally fed preterm infants is unknown. We hypothesized that preterm infants show a high splanchnic first-pass glutamine metabolism and the primary metabolic fate of glutamine is oxidation. Five preterm infants (mean + or - SD birth weight 1.07 + or - 0.22 kg and GA 29 + or - 2 wk) were studied by dual tracer ([U (13)C]glutamine and [(15)N(2)]glutamine) cross-over techniques on two study days (at postnatal week 3 + or - 1 wk). Splanchnic and whole-body glutamine kinetics were assessed by plasma isotopic enrichment of [U-(13)C]glutamine and [(15)N(2)]glutamine and breath (13)CO(2) enrichments. Mean fractional first-pass glutamine uptake was 73 + or - 6% and 57 + or - 17% on the study days. The splanchnic tissues contributed for a large part (57 + or - 6%) to the total amount of labeled carbon from glutamine retrieved in expiratory air. Dietary glutamine is used to a great extent by the splanchnic tissues in preterm infants and its carbon skeleton has an important role as fuel source. PMID- 19809374 TI - Urotensin-II contributes to pulmonary vasoconstriction in a perinatal model of persistent pulmonary hypertension of the newborn secondary to meconium aspiration syndrome. AB - Meconium aspiration syndrome (MAS) disrupts perinatal decreases in pulmonary vascular resistance (PVR) and is the commonest cause of neonatal pulmonary hypertension. The contribution of the potent vasoactive agent urotensin-II (U II), in the pathophysiology of this condition, is unknown. In a new perinatal model of MAS, we combined measurement of circulating U-II levels with U-II receptor blockade studies. Nineteen anesthetized lambs were instrumented then randomly allocated to the following groups: 1) control (n = 5), 2) control plus specific U-II receptor blockade with palosuran (n = 5), 3) tracheal instillation of meconium (n = 5), 4) meconium instillation plus palosuran (n = 4). Hemodynamics, PVR, and plasma U-II were measured for 6 h after delivery. After birth in controls, U-II increased (p < 0.05), and PVR fell (p = 0.01) and this fall was prevented by U-II receptor blockade. By contrast, meconium lambs displayed a greater rise in U-II levels (p < 0.05 versus control) with an increase in PVR (p < 0.005) that was attenuated by U-II receptor blockade (p < 0.001). These findings suggest that U-II normally acts as a pulmonary vasodilator after birth, but in the presence of MAS, it assumes a vasoconstrictor role. U-II receptor blockade also improves pulmonary hemodynamics in this model. PMID- 19809375 TI - Thioredoxin binding protein-2 inhibits excessive fetal hypoglycemia during maternal starvation by suppressing insulin secretion in mice. AB - Glucose is a major fuel for fetal development. Fetal blood glucose level is mainly dependent on maternal blood glucose concentration, though it is also regulated by fetal insulin level. Thioredoxin binding protein-2 (TBP-2), which is identical to vitamin D3 up-regulated protein (VDUP1) and thioredoxin interacting protein (Txnip), was recently reported to be a key transcriptional factor controlling glucose metabolism. Here, we elucidated the functions of TBP-2 in maintaining blood glucose homeostasis during the fetal period. TBP-2(+/-) female mice were mated with TBP-2(+/-) male mice; beginning 16.5-d post coitum, pregnant mice were fed or fasted for 24 h. Under conditions of maternal starvation, the blood glucose levels of TBP-2(-/-) fetuses were significantly lower than those of TBP-2(+/+) fetuses, corresponding to the elevated plasma insulin levels of TBP-2( /-) fetuses compared with those of TBP-2(+/+) fetuses. There was no difference between TBP-2(+/+) and TBP-2(-/-) fetuses in terms of their pancreatic beta-cell masses or the expression of placental glucose transporters under conditions of either maternal feeding or fasting. Thus, during maternal fasting, fetal TBP-2 suppresses excessive insulin secretion to maintain the fetus's glucose levels, implying that TBP-2 is a critical molecule in mediating fetal glucose homeostasis depending on nutrient availability. PMID- 19809376 TI - Relationship of proximal renal tubular dysgenesis and fetal liver injury in neonatal hemochromatosis. AB - Renal tubular dysgenesis has been reported in isolated cases of neonatal hemochromatosis (NH). We hypothesized that fetal liver injury in NH impairs proximal renal tubular development via impaired hepatic angiotensinogen (AGT) elaboration. Morphometric analyses were performed of postmortem liver and kidney sections of cases of proven NH and postconception age-matched controls for renal proximal tubule density, hepatocyte mass, and hepatic AGT expression. Proximal tubule density was markedly reduced in NH cases, although they showed a spectrum from mild to severe paucity. Hepatic AGT expression was markedly reduced in NH cases and correlated closely with reduced hepatocyte mass. A linear relationship was established between hepatic AGT expression and the degree of renal tubular dysgenesis suggesting that there is a relationship between them. Our results demonstrate that there is a spectrum of kidney pathology in patients with NH including a large proportion of cases with severe proximal tubular dysgenesis. Hepatic synthetic failure resulting in insufficient production of AGT to support renal tubular development is the likely mechanism of kidney disease in NH. PMID- 19809377 TI - Hyperoxia exposure alters hepatic eicosanoid metabolism in newborn mice. AB - Prematurely born infants are often treated with supraphysiologic amounts of oxygen, which is associated with lung injury and the development of diseases such as bronchopulmonary dysplasia. Complimentary responses between the lung and liver during the course of hyperoxic lung injury have been studied in adult animals, but little is known about this relationship in neonates. These studies tested the hypothesis that oxidant stress occurs in the livers of newborn mice in response to continuous hyperoxia exposure. Greater levels of glutathione disulfide and nitrotyrosine were detected in lung tissues but not liver tissues from newborn mice exposed to hyperoxia than in room air-exposed controls. However, early increases in 5-lipoxygenase and cyclooxygenases-2 protein levels and increases in total hydroxyeicosatetraenoic acid and prostaglandin levels were observed in the liver tissues of hyperoxia-exposed pups. These studies indicate that free radical oxidation occurs in the lungs of newborn pups exposed to hyperoxia, and alterations in lipid metabolism could be a primary response in the liver tissues. The findings of this study identify possible new mechanisms associated with hyperoxic lung injury in a newborn model of bronchopulmonary dysplasia and thus open opportunities for research. PMID- 19809378 TI - In vivo dilatation of the ductus arteriosus induced by furosemide in the rat. AB - Furosemide increases prostaglandin production and may be associated with patent ductus arteriosus (PDA). We aimed to clarify the in vivo ductus-dilating effects of furosemide in neonatal rats. Near-term rat pups delivered by a cesarean section were housed at 33 degrees C. After a rapid whole-body freezing, the DA diameter was measured using a microscope and a micrometer. Pregnant rats (gestational day 21) were s.c. injected with furosemide 4 h before delivery, and the neonatal DA was examined 0, 15, 30, 60, and 120 min after birth. Furosemide was also s.c. injected into 60-min-old rats and the DA diameter was examined 30, 60, and 120 min later. The control rats showed a rapid postnatal DA constriction (diameter: 0.80 and 0.08 mm at 0 and 60 min after birth, respectively). Prenatally administered furosemide delayed postnatal DA closure (0.36 mm at 60 min after birth). Furosemide injection in 60-min-old rats dilated the constricted DA at 60 min (0.25 versus 0.02 mm in the controls). Indomethacin inhibited furosemide-induced DA dilatation. Furosemide delays DA closure and dilates the constricted DA in neonatal rats. If furosemide has similar effects in human preterm neonates, caution may be warranted in its use in the treatment of infants with PDA. PMID- 19809379 TI - ATPase Class I Type 8B Member 1 and protein kinase C zeta induce the expression of the canalicular bile salt export pump in human hepatocytes. AB - The exact molecular mechanism(s) of the disease that results from defects in the ATPase Class I Type 8B Member 1 gene remains controversial. Prior investigations of human ileum and in intestinal and ovarian cell lines have suggested that familial intrahepatic cholestasis 1 (FIC1) activates the farnesoid X-receptor (FXR) via a pathway involving protein kinase C zeta (PKCzeta). Translational investigations of human liver from individuals with FIC1 disease have been confounded by secondary affects of progressive cholestatic liver disease and limited numbers of samples for analysis. These studies, performed in primarily derived human hepatocytes, circumvent this issue. The canalicular bile salt export pump (BSEP) served as a downstream target of FXR. The siRNA-mediated silencing of FIC1 in human hepatocytes led to a reduction in both human BSEP promoter activity and BSEP protein expression, which correlated with a reduction in FXR expression and redistribution of its localization from the nucleus to the cytoplasm. These changes in BSEP expression could be reproduced by altering the expression of PKCzeta, with a positive correlation of PKCzeta activity and BSEP expression. Overall, these findings support the hypothesis that FIC1 enhances FXR signaling via a PKCzeta-dependent signaling pathway. PMID- 19809380 TI - A model for educational simulation of hemodynamic transitions at birth. AB - Birth is characterized by swift and complex transitions in hemodynamic and respiratory variables. Unrecognized pathologies or incidents may quickly become fatal or cause permanent damage. This article introduces an essential component of an acute perinatal care simulator, namely a model for educational simulation of normal hemodynamic transitions seen during and shortly after birth. We explicitly formulate educational objectives and adapt a preexisting model for the simulation of neonatal cardiovascular physiology to include essential aspects of fetal hemodynamics. From the scientific literature, we obtain model parameters that characterize these aspects quantitatively. The fetal model is controlled by a time- and event-based script of changes occurring at birth, such as onset of breathing and cord clamping, and the transitory phase up to 24 h after birth. Comparison of simulation results with published target data confirms that realistic simulated hemodynamic vital signs are achieved. PMID- 19809381 TI - Fetal programming of infant neuromotor development: the generation R study. AB - The objective of the study was to examine whether infant neuromotor development is determined by fetal size and body symmetry in the general population. This study was embedded within the Generation R Study, a population-based cohort in Rotterdam. In 2965 fetuses, growth parameters were measured in mid-pregnancy and late pregnancy. After birth, at age 9 to 15 wks, neuromotor development was assessed with an adapted version of Touwen's Neurodevelopmental Examination. Less optimal neuromotor development was defined as a score in the highest tertile. We found that higher fetal weight was beneficial to infant neurodevelopment. A fetus with a 1-SD score higher weight in mid-pregnancy had an 11% lower risk of less optimal neuromotor development (OR: 0.89; 95% CI: 0.82-0.97). Similarly, a fetus with a 1-SD score larger abdominal-to-head circumference (AC/HC) ratio had a 13% lower risk of less optimal neuromotor development (OR: 0.87; 95% CI: 0.79-0.96). These associations were also present in late pregnancy. Our findings show that fetal size and body symmetry in pregnancy are associated with infant neuromotor development. These results suggest that differences in infant neuromotor development, a marker of behavioral and cognitive problems, are at least partly caused by processes occurring early in fetal life. PMID- 19809382 TI - Post-treatment tumor gene expression signatures are more predictive of treatment outcomes than baseline signatures in breast cancer. AB - OBJECTIVE: Tumor gene expression signatures have been used to classify, prognosticate, and predict chemotherapy sensitivity in breast cancer, although almost all efforts have been focused on the unchallenged baseline tumor. Most cancer patients receive systemic therapy, and exposure to drug may modify the tumor's short-term and long-term outcomes. Drug-induced tumor gene signatures may thus be more predictive of treatment outcomes than the unperturbed tumor gene signatures. METHODS: Using a set of 47 breast cancer patients, we obtained paired prechemotherapy and postchemotherapy tumor biopsies and developed gene panels of baseline tumor (T1), postchemotherapy tumor (T2), and chemotherapy-induced relative change signatures (TDelta) to predict pathological response and progression-free survival (PFS). The signatures were validated in two independent test sets with paired prechemotherapy and postchemotherapy tumor samples, comprising of 18-20 patients each. RESULTS: T2 and TDelta were superior to T1 signatures in predicting for PFS (area under the curve of receiver operating characteristic 0.770 and 0.660 vs. 0.530) and pathological response (area under the curve of receiver operating characteristic 0.631 and 0.462 vs. 0.446) in the validation sets. In multivariate analysis for PFS with other clinical predictors, T2, but not T1, signatures remained as significant independent predictors. CONCLUSION: Postchemotherapy tumor gene signatures outperformed baseline signatures and clinical predictors in predicting for pathological response and PFS, independent of clinical and pathological response to chemotherapy. Drug induced tumor gene signatures may be more informative than unchallenged signatures in predicting treatment outcomes. These findings challenge the current practice of relying only on the baseline tumor to predict outcome, which overlooks the contributions of therapeutic interventions. PMID- 19809383 TI - Screening for HSV-2 infection in STD clinics and beyond: a few answers but more questions. PMID- 19809384 TI - Implementation of routine access to herpes simplex virus type 2 antibody testing in a public health sexually transmitted disease clinic. AB - BACKGROUND: Testing for herpes simplex virus type 2 (HSV-2) antibody is not common in clinical practice. Client characteristics associated with HSV-2 rapid antibody test uptake and test positivity were analyzed in clients attending an urban sexually transmitted disease clinic. METHODS: This optional test was available for $30. The HerpeSelect Express assay was performed on serum. Demographic and behavioral characteristics were compared between clients who requested testing and those who did not and between those who were HSV-2 antibody positive and negative. RESULTS: In 4 months, 3498 individuals attended the clinic and 443 (12.7%) opted for HSV-2 testing. Clients who were black, younger, or female were less likely to request testing. Recent sexual behavior and self reported sexual orientation were not associated with uptake of testing. Of the 442 clients with results available, 109 were positive for HSV-2 antibody (24.7%). Women were significantly (P <0.001) more likely to test positive; 42 of 111 (38.4%) versus only 67 of 331 (20.2%) men. A positive HSV-2 antibody test was also associated with increasing age and black race. There was an association with the number of partners in the last 30 days, but no association with the number of partners in the last year. Of the 109 clients who had a positive HSV-2 antibody test, 71 (64.5%) accepted a prescription for suppressive acyclovir therapy. CONCLUSIONS: Uptake of testing was modest in this population, especially among the highest risk individuals, possibly due to the cost of the test. Improved education regarding HSV-2 and subsidized testing may be needed in the populations that have the highest prevalence in order to encourage testing. PMID- 19809385 TI - HPV 6/11, 16, 18 seroprevalence in men in two US cities. AB - BACKGROUND: A vaccine to prevent human papillomavirus (HPV) 6, HPV 11, HPV 16, or HPV 18 and associated diseases is licensed for females, and it may be licensed for men in the future. There are limited data on HPV 6/11, 16, and/or 18 seroprevalence in men. METHODS: A total of 490 men aged 18 to 40 years were enrolled in a study of HPV in men in Tucson, AZ, and Tampa, FL. Enrolled men completed a self-administered questionnaire, and HPV serology was performed using HPV 6/11, 16, and 18 VLP assays. RESULTS: Overall, seroprevalence to HPV 16 was 12.1%, HPV 6/11 was 9.7%, and to HPV 18 was 5.4%. Seroprevalence to HPV 6/11, 16, and/or 18 was 21% and was highest among 35 to 40 year olds (48%); prevalence in this age group was significantly higher compared to the 18 to 24 year olds (adjusted odds ratio (aOR) 6.8, 95% confidence interval (CI) 3.7, 12.8). Independent predictors of seropositivity to HPV 6/11, 16, and/or 18 were older age, greater number of female sex partners in the past 3 months, and current smoking. CONCLUSIONS: HPV vaccine-type seroprevalence was highest in 35 to 40 year old men. These data on the epidemiology of HPV seroprevalence in men are useful for discussions regarding recommendations for HPV vaccine if licensed for use in men. PMID- 19809386 TI - Testosterone concentrations in hair of hypogonadal men with and without testosterone replacement therapy. AB - BACKGROUND: The measurement of testosterone (T) is essential for the diagnosis of male hypogonadism and for monitoring treatment. Samples need to be obtained at specific times in relation to the diurnal rhythm and therapeutic T injections. In this study, we explored the measurement of T in hair as an alternative method to assess gonadal status and long-term T exposure in men. SUBJECTS AND METHODS: Thirty-six male subjects comprising 17 healthy volunteers, 10 untreated hypogonadal men, and nine hypogonadal men receiving T injections were studied. T was measured in serum and in hair. T in hair was measured using a commercially available salivary T enzyme immunoassay kit adapted for this use. RESULTS: The T concentration in the hair of hypogonadal men receiving T injections was significantly higher than that in untreated hypogonadal volunteers, but not eugonadal men. Median T concentrations were 3.66 (range, 0.82-15.00), 0.94 (range, 0.33-3.68), and 1.85 (range, 0.58-3.06) pg/g hair, respectively. CONCLUSIONS: T in hair reflects gonadal status in men and may be useful for monitoring T therapy over several weeks to months in hypogonadal men. PMID- 19809387 TI - Vancomycin continuous infusion as prophylaxis for vascular surgery. AB - Prosthetic graft infection is a devastating complication of vascular surgery that occurs in 3%-5% of clean prosthetic procedures. Staphylococci are the most frequently isolated pathogens, and thus surgical prophylaxis regimens often include vancomycin. However, the efficacy of these regimens in ensuring a required concentration of antibiotic is uncertain. This study aimed to determine if a continuous vancomycin infusion regimen administered perioperatively as surgical prophylaxis for vascular procedures maintained an adequate serum concentration. Thirty-four consecutive patients undergoing a vascular procedure requiring a prosthetic graft or patch were given vancomycin prophylaxis. Each patient received a loading dose calculated according to body weight 12 hours before surgery. A 24-hour continuous infusion was then started, based on calculated creatinine clearance. Serum vancomycin concentrations were checked on induction of anesthesia, 2 hours postoperatively, and at the end of the infusion. Perioperative fluid administration and blood loss were recorded. An estimated creatinine clearance was repeated on the second postoperative day. Of the 34 patients recruited, 7 did not have the anticipated procedure and 6 patients had incomplete sample collection. Twenty-one patients with complete sample collection were analyzed. The target concentration (10-25 mg/L) was achieved in 81% of all samples. All patients achieved the target concentration at 1 or more time points. The regimen employed provided appropriate concentrations at the time of intervention. No potentially toxic concentrations or adverse reactions to vancomycin were encountered. Vancomycin given as a continuous infusion delivers adequate serum concentration. Long-term graft infection rates are needed to show a clinical effect. PMID- 19809388 TI - High-performance liquid chromatography using UV detection for the simultaneous quantification of ropivacaine and bupivacaine in human plasma. AB - A specific high-performance liquid chromatography assay coupled with UV detection has been developed and validated for the simultaneous determination of ropivacaine and bupivacaine in human plasma. A liquid-liquid back extraction procedure was used to increase specificity, and very good and consistent recoveries were obtained: 93%-95% for ropivacaine and 90%-96% for bupivacaine. The lowest limit of quantification was 4 and 8 ng/mL for ropivacaine and bupivacaine, respectively. The method was sensitive, reproducible (coefficient of variation 207 for MPA, 438-->303 for MPAC, and 514-->303 for MPAG and AcMPAG. The dynamic ranges (lower limit of quantitation and upper limit of quantitation) were as follows: 0.05 to 30 mg/L for total MPA and 1 to 300 microg/L for free MPA; 0.5 to 300 mg/L of total MPAG and 0.2 to 60 mg/L for free MPAG; and 0.025 to 15 mg/L of total AcMPAG and 1 to 60 microg/L for free AcMPAG. The precision at lower limit of quantitation was in the range of 8.0% to 11.9% for all three total analytes and 13.8 to 18.7% for the free analytes. Accuracy at lower limit of quantitation was in the range of 100% to 105% for total and 97% to 99% for free analytes. Between-day precision of quality control samples was 4.0% to 6.3% for human plasma spiked with total analytes and 4.5% to 14.4% for spiked plasma ultrafiltrate for free analytes. Mean absolute recovery ranged from 98.5% to 101.7% for MPA (both total and free), from 78.1% to 103.4% for MPAG and from 91.5% to 110.4% for AcMPAG. No significant ion suppression was found under these conditions for any of the analytes. Carryover effect was found to be at a maximum level of 0.02%. This method was successfully applied to analyze over 11,000 samples for total analytes, and over 8000 samples for free analytes in plasma, and has been in operation for nearly 3 years without loss of performance. PMID- 19809390 TI - Affirming the fundamentals in an age of high-tech rehabilitation. PMID- 19809391 TI - Treadmill-based locomotor training with leg weights to enhance functional ambulation in people with chronic stroke: a pilot study. AB - BACKGROUND AND PURPOSE: Novel locomotor training strategies for individuals with disorders of the central nervous system have been associated with improved locomotor function. The purpose of this study was to investigate the effects of treadmill-based locomotor training combined with leg weights on functional ambulation in individuals with chronic stroke. We assessed functional ambulation and muscle activity in ambulatory individuals with chronic stroke. METHODS: We used a pre/posttest design. Six individuals with chronic stroke who were community ambulators were recruited. Participants underwent a 30-minute treadmill based locomotor training sessions three times per week for four to 12 weeks. The training program involved treadmill walking for 30 minutes with partial body weight support as needed. Leg weights, equivalent to 5% of body weight, were affixed around the paretic leg. Outcome measures consisted of the 10-m walk test, the modified Emory Functional Ambulation Profile, and temporal gait parameters. RESULTS: Improvements were observed in functional ambulation measures, particularly the stairs subscore of the modified Emory Functional Ambulation Profile. Participants also exhibited an increase in the proportion of time the paretic leg spent in swing. No significant improvements were observed in the 10-m walk test. CONCLUSIONS: This pilot study demonstrates that the combination of leg weights and treadmill training is a feasible approach, that is well tolerated by participants. This approach may have the potential to improve some aspects of functional ambulation and the performance of activities requiring hip and knee flexion. PMID- 19809392 TI - The accuracy of individual Berg Balance Scale items compared with the total Berg score for classifying people with chronic stroke according to fall history. AB - BACKGROUND AND PURPOSE: To determine whether individual Berg Balance Scale (BBS) items or a group of items would have greater accuracy than the total BBS in classifying community-dwelling people with stroke with a history of multiple falls. METHODS: The subjects were 44 community-dwelling individuals with chronic stroke; 34 had one or no falls in the past six months, and 10 had multiple falls. Each BBS item was dichotomized at three points along the scoring scale of 0-4: between scores of 1 and 2, 2 and 3, and 3 and 4. Sensitivity (Sn), specificity (Sp), and positive (+LR) and negative (-LR) likelihood ratios were calculated for all items for each scoring dichotomy based on their accuracy in classifying subjects with a history of multiple falls. These findings were compared with the total BBS score where the cutoff score was derived from receiver operating characteristic curve analysis. RESULTS: Dichotomized point 3-4 for items B11 (turning 360 degrees), B12 (alternate foot on stool), B13 (tandem stance), and B14 (standing on one leg) all revealed Sn greater than 60%. B14 had the best Sn and Sp (0.90 and 0.50). Combining B11, B12, or B13 with B14 did not improve Sn. Total BBS receiver operating characteristic curve revealed a cutoff score of 52 (Sn = 90% and Sp = 41%). CONCLUSION: Using selected items from the BBS may be more time efficient and accurate than the total BBS score for classifying people with chronic stroke living in the community with a history of multiple falls. Prospective study is needed to validate these findings relative to fall prediction. PMID- 19809393 TI - Characterizing gait, locomotor status, and disease severity in children and adolescents with Friedreich ataxia. AB - BACKGROUND AND PURPOSE: The purpose of this study was to describe gait parameters in children and adolescents with a diagnosis of Friedreich ataxia (FA) and examine the relationship between disease severity, measured by the Friedreich Ataxia Rating Scale (FARS) and gait parameters. The study examined whether FARS scores can discriminate between those who walk independently and those who require assistance. METHODS: Thirty-eight children (aged 5-11 years) and adolescents (aged 12-17 years) with genetically confirmed FA were divided into two groups based on locomotor status: group 1, subjects who were able to walk independently, and group 2, subjects who required assistance for walking. Temporal and spatial gait parameters were collected using the Stride Analyzer computerized foot switch system and compared with age-matched normative data. The FARS was used to measure disease severity. Correlation coefficients and the Mann Whitney U test of differences were used to evaluate associations and discern differences between groups. RESULTS: In subjects with FA, gait parameters of velocity and cadence were slower and stride length was shorter compared with age matched children without disabilities. These parameters were significantly correlated with FARS score (r = 0.696, 0.667, 0.537; respectively, all P values <0.001). Total FARS scores were correlated with locomotor status (c value r = 0.623; P < 0.01) and could categorize subjects into groups based on independent walking or need for assistance, 73% and 87% of the time, respectively. DISCUSSION AND CONCLUSION: Subjects with FA exhibited specific abnormal gait characteristics relative to age-matched individuals. Disease severity, as measured by the FARS, was associated with gait velocity, stride length, and cadence. FARS scores can be used to categorize subjects by locomotor status and may be a useful screening tool to identify those requiring assistance. PMID- 19809394 TI - Use of an electrotactile vestibular substitution system to facilitate balance and gait of an individual with gentamicin-induced bilateral vestibular hypofunction and bilateral transtibial amputation. AB - BACKGROUND AND PURPOSE: This case report describes the use of an electrotactile vestibular substitution system (ETVSS; BrainPort Balance Device, Wicab, Inc., Middleton, WI) to facilitate balance and gait of an individual with bilateral vestibular hypofunction and bilateral transtibial amputation. CASE DESCRIPTION: A 69-year-old man with a 2.5-year history of bilateral vestibular hypofunction, due to gentamicin toxicity, participated in a rehabilitation program using an ETVSS. Because of lower extremity infection, the patient had bilateral prosthetic legs after bilateral transtibial amputation. INTERVENTION: Focused on the following three phases of training with the ETVSS during 12 months: orientation, clinical training, and in-home training. The patient was periodically assessed with balance and gait tests, in addition to surveys of patient confidence and perception of handicap. All testing was performed without ETVSS. OUTCOMES: : Improvements were demonstrated in all outcome measures used with this patient. Sensory Organization Test composite scores increased from 23 to 48, Dynamic Gait Index scores increased from 11/24 to 21/24, and distance walked during six minutes increased from 212 to 363 m. Standing balance with eyes-closed improved from less than two seconds to more than 20 minutes. The patient reported improved confidence and lower perception of handicap with fewer functional limitations. DISCUSSION: For this patient, an intervention program of sensory substitution using the ETVSS improved outcome measures beyond those previously achieved with vestibular rehabilitation therapy and balance training. The feedback provided by the ETVSS may have facilitated the patient's ability to use proprioception, thus allowing better balance control. PMID- 19809395 TI - Challenge-oriented gait and balance training in sporadic olivopontocerebellar atrophy: a case study. AB - BACKGROUND AND PURPOSE: Sporadic olivopontocerebellar atrophy (OPCA) is a rare and debilitating neurologic disease of insidious onset. It is characterized by atrophy of the cerebellum, pons, and inferior olivary nuclei with concomitant ambulation deficits and dyscoordination. To our knowledge, there has been no published study investigating any aspect of rehabilitation in OPCA. Therefore, the purpose of this study was to investigate the use of challenge-oriented gait and balance training to improve gait and balance in OPCA. CASE DESCRIPTION: An otherwise healthy 19-year-old woman with moderate to severe upper and lower extremity ataxia, secondary to sporadic OPCA, participated in this prospective case study. She also had a vestibulotoxic treatment procedure to decrease the severity of her vertigo. INTERVENTION: This individual participated in a 12-week gait and balance training program (five times per week), which consisted of one to two hours of various challenging static and dynamic balance tasks. To measure her progress, the following scales and tests were used: Berg Balance Scale, Dynamic Gait Index, Activities-Specific Balance Confidence Scale, computerized dynamic posturography (sensory organization test and limits of stability), and self-selected gait velocity. OUTCOMES: : Improvements were noted in all the dependent measures (pre to post): Berg Balance Scale (34/56 to 39/56), Dynamic Gait Index (1/24 to 7/24), Activities-Specific Balance Confidence Scale (50.6% 85.1%), sensory organization test (composite score, 31/100 to 47/100), limits of stability (maximum excursion, 89-105; endpoint excursion, 57-93; directional control, 60-78), and SSGV (0.375-0.526 m/sec). DISCUSSION: Results from this case study suggest that a gait and balance training program may be beneficial to individuals with ataxia from OPCA. This early evidence warrants further investigation using more rigorous methods. PMID- 19809396 TI - President's perspectives. Healthcare Reform: a call to action for our patients. PMID- 19809400 TI - Mouse Mammary Epithelial Cells form Mammospheres During Lactogenic Differentiation. AB - A phenotypic measure commonly used to determine the degree of lactogenic differentiation in mouse mammary epithelial cell cultures is the formation of dome shaped cell structures referred to as mammospheres. The HC11 cell line has been employed as a model system for the study of regulation of mammary lactogenic differentiation both in vitro and in vivo. The HC11 cells differentiate and synthesize milk proteins in response to treatment with lactogenic hormones. Following the growth of HC11 mouse mammary epithelial cells to confluence, lactogenic differentiation was induced by the addition of a combination of lactogenic hormones including dexamethasone, insulin, and prolactin, referred to as DIP. The HC11 cells induced to differentiate were photographed at times up to 120 hours post induction of differentiation and the number of mammospheres that appeared in each culture was enumerated. The size of the individual mammospheres correlates with the degree of differentiation and this is depicted in the images of the differentiating cells. PMID- 19809401 TI - A comparison of the growth responses following intramuscular GHRH plasmid administration versus daily growth hormone injections in young pigs. AB - The efficacy of daily porcine growth hormone (GH) injections versus plasmid driven porcine GH-releasing hormone (pGHRH) production to promote growth was assessed. Ten-day-old piglets were injected intramuscularly with 0.1, 1, or 3 mg pGHRH, or a control plasmid followed by electroporation. Plasmid constructs were driven by a synthetic muscle-specific promoter. A fifth group received daily injections of GH [0.15 mg/(kg.day)]. Control and pGHRH-treated pigs were pair-fed to GH-treated pigs. Body composition was assessed by dual-energy X-ray absorptiometry (DXA). Weight gains of GH- and pGHRH-treated pigs were greater than of controls (P < 0.001) due to greater lean mass accretion; fat accretion was similar across all treatments. Weight gain of pGHRH- and GH-treated pigs was similar for 6 weeks, but over the final 10 days, only pigs administered the highest plasmid dose maintained higher growth rates. Serum insulin-like growth factor-I (IGF-I) levels were two- to threefold higher in GH- and pGHRH-treated pigs than in controls after 4 weeks (P = 0.05), but subsequently decreased to control levels in the pGHRH-treated group. Organ weights were greater in GH- than pGHRH-treated and control piglets (P < 0.02). These results demonstrate that pGHRH transfer is effective for promoting growth and avoids the need for the frequent injections necessitated with peptide hormone use. PMID- 19809402 TI - Combinatorial control of suicide gene expression by tissue-specific promoter and microRNA regulation for cancer therapy. AB - Transcriptional targeting using a tissue-specific cellular promoter is proving to be a powerful means for restricting transgene expression in targeted tissues. In the context of cancer suicide gene therapy, this approach may lead to cytotoxic effects in both cancer and nontarget normal cells. Considering microRNA (miRNA) function in post-transcriptional regulation of gene expression, we have developed a viral vector platform combining cellular promoter-based transcriptional targeting with miRNA regulation for a glioma suicide gene therapy in the mouse brain. The therapy employed, in a single baculoviral vector, a glial fibrillary acidic protein (GFAP) gene promoter and the repeated target sequences of three miRNAs that are enriched in astrocytes but downregulated in glioblastoma cells to control the expression of the herpes simplex virus thymidine kinase (HSVtk) gene. This resulted in significantly improved in vivo selectivity over the use of a control vector without miRNA regulation, enabling effective elimination of human glioma xenografts while producing negligible toxic effects on normal astrocytes. Thus, incorporating miRNA regulation into a transcriptional targeting vector adds an extra layer of security to prevent off-target transgene expression and should be useful for the development of gene delivery vectors with high targeting specificity for cancer therapy. PMID- 19809403 TI - PiggyBac transposon-based inducible gene expression in vivo after somatic cell gene transfer. AB - Somatic cell gene transfer has permitted inducible gene expression in vivo through coinfection of multiple viruses. We hypothesized that the highly efficient plasmid-based piggyBac transposon system would enable long-term inducible gene expression in mice in vivo. We used a multiple-transposon delivery strategy to create a tetracycline-inducible expression system in vitro in human cells by delivering the two genes on separate transposons for inducible reporter gene expression along with a separate selectable transposon marker. Evaluation of stable cell lines revealed 100% of selected clones exhibited inducible expression via stable expression from three separate transposons simultaneously. We next tested and found that piggyBac-mediated gene transfer to liver or lung could achieve stable reporter gene expression in mice in vivo in either immunocompetent or immune deficient animals. A single injection of piggyBac transposons could achieve long-term inducible gene expression in the livers of mice in vivo, confirming our multiple-transposon strategy used in cultured cells. The plasmid based piggyBac transposon system enables constitutive or inducible gene expression in vivo for potential therapeutic and biological applications without using viral vectors. PMID- 19809404 TI - Engineered newcastle disease virus as an improved oncolytic agent against hepatocellular carcinoma. AB - Newcastle disease virus (NDV) is an intrinsically tumor-specific virus, which is currently under investigation as a clinical oncolytic agent. Several clinical trials have reported NDV to be a safe and effective agent for cancer therapy; however, there remains a clear need for improvement in therapeutic outcome. The endogenous NDV fusion (F) protein directs membrane fusion, which is required for virus entry and cell-cell fusion. Here, we report a novel NDV vector harboring an L289A mutation within the F gene, which resulted in enhanced fusion and cytotoxicity of hepatocellular carcinoma (HCC) cells in vitro, as compared with the rNDV/F3aa control virus. In vivo administration of the recombinant vector, termed rNDV/F3aa(L289A), via hepatic arterial infusion in immune-competent Buffalo rats bearing multifocal, orthotopic liver tumors resulted in tumor specific syncytia formation and necrosis, with no evidence of toxicity to the neighboring hepatic parenchyma. Furthermore, the improved oncolysis conferred by the L289A mutation translated to significantly prolonged survival compared with control NDV. Taken together, rNDV/F(L289A) represents a safe, yet more effective vector than wild-type NDV for the treatment of HCC, making it an ideal candidate for clinical application in HCC patients. PMID- 19809405 TI - Variability of histopathological changes in childhood celiac disease. AB - OBJECTIVES: Adult studies of celiac disease (CD) have shown that duodenal mucosal histopathological changes may be patchy, and the diagnostic utility of duodenal bulb biopsies is believed to be limited. Few related pediatric data exist. METHODS: We assessed the prevalence of variable biopsy findings and duodenal bulb involvement in children with CD, as well as its association with clinical parameters. A total of 198 consecutive cases of CD diagnosed at the Children's Hospital during 2001-2005 were analyzed. All biopsies were scored by a pathologist blinded to the clinical data using the Marsh criteria. Mucosal changes were classified as focal if changes consistent with CD and normal mucosa were found within a single biopsy fragment. Patchiness was defined as variation of at least one Marsh grade between separate fragments in a biopsy set. RESULTS: The median age was 9.3 years; 62% were female. An average of 3.6 biopsy samples was obtained per case. In 101 cases, biopsy samples were obtained from the duodenal bulb and the second portion of the duodenum. Focality was present in biopsy samples collected from 36 (18%) cases. Patchiness was found in 105 (53%) cases, and at least 1 normal biopsy fragment was present in 71 (36%) cases. In 10 cases, only the bulb biopsies were diagnostic of CD. There was no association with the clinical features examined. CONCLUSIONS: Duodenal involvement in pediatric CD is frequently patchy and may show variable severity even within a single biopsy fragment. Variability cannot be predicted by clinical characteristics. Multiple endoscopic biopsies, including the duodenal bulb, should be obtained in suspected pediatric CD cases to maximize diagnostic yield. PMID- 19809406 TI - Increased prevalence of and associated mortality with methicillin-resistant Staphylococcus aureus among hospitalized IBD patients. AB - OBJECTIVES: Methicillin-resistant Staphylococcus aureus (MRSA) infection has become increasingly prevalent in US hospitals, and the impact of MRSA on hospitalized inflammatory bowel disease (IBD) patients is unknown. METHODS: We used the Nationwide Inpatient Sample to identify admissions for IBD (n=116,842) between 1998 and 2004. We compared prevalence and in-hospital mortality of MRSA among IBD, non-IBD gastrointestinal (GI), and general medical inpatients. RESULTS: MRSA prevalence increased from 4.5/10,000 to 19.0/10,000 over the 7-year period (P<0.0001). After adjustment for confounders, IBD inpatients were at increased risk of MRSA compared with the non-IBD GI (adjusted odds ratio (aOR) 1.61; 95% confidence interval (CI): 1.33-1.96) and general medical (aOR 1.36; 95% CI: 1.11-1.66) groups. Of those with MRSA, catheter-related infections were specifically more common among IBD compared with non-IBD GI and general inpatients (28.8% vs. 11.0% and 8.5%, respectively, P<0.0002). Bowel surgery, parenteral nutrition, and health insurance were predictors of MRSA infection, but the first two became insignificant after controlling for length of stay (LOS). Compared with LOS < or = 7 days, MRSA was more likely among those hospitalized 8 21 days (aOR 7.40; 95% CI: 4.68-11.7) and >21 days (aOR 58.6; 95% CI: 36.0-95.3). MRSA infection was associated with sevenfold increase in mortality (aOR 7.61; 95% CI: 3.33-17.4). CONCLUSIONS: Hospitalized IBD patients are at increased risk of MRSA compared with non-IBD GI and general medical inpatients. Increased mortality in the IBD population associated with MRSA reinforces the importance of measures to prevent nosocomial infection and to reduce length of hospitalization. PMID- 19809407 TI - Negative lymph node count is associated with survival of colorectal cancer patients, independent of tumoral molecular alterations and lymphocytic reaction. AB - OBJECTIVES: The number of recovered lymph nodes is associated with good prognosis among colon cancer patients undergoing surgical resection. However, little has been known on prognostic significance of lymph node count after adjusting for host immune response to tumor and tumoral molecular alterations, both of which are associated with the lymph node count and patient survival. METHODS: Among 716 colorectal cancers (stages 1-4) in two independent prospective cohorts, we examined patient survival in relation to the negative lymph node count and lymph node ratio (LNR; positive to total lymph node counts). Cox proportional hazard models were used to compute hazard ratio of deaths, adjusted for patient, specimen, and tumoral characteristics, including lymphocytic reactions, KRAS and BRAF mutations, p53 expression, microsatellite instability (MSI), the CpG island methylator phenotype (CIMP), and LINE-1 methylation. RESULTS: Compared with patients with 0-3 negative lymph nodes, patients with 7-12 and > or =13 negative nodes experienced a significant reduction in cancer-specific and overall mortality in Kaplan-Meier analysis (log-rank P<0.0001), univariate Cox regression (P(trend)<0.0001), and multivariate analysis (P(trend)<0.0003), independent of potential confounders examined. The benefit associated with the negative node count was apparent across all stages, although the effect was significantly greater in stages 1-2 than stages 3-4 (P(interaction)=0.002). In both stage 3 and stage 4, smaller LNR was associated with improved survival (log-rank P<0.0001). CONCLUSIONS: The negative lymph node count is associated with improved survival of colorectal cancer patients, independent of lymphocytic reactions to tumor and tumoral molecular features including MSI, CIMP, LINE-1 hypomethylation and BRAF mutation. PMID- 19809408 TI - A randomized, double-blind, placebo-controlled trial of St John's wort for treating irritable bowel syndrome. AB - OBJECTIVES: St John's wort (SJW) is known to effectively treat patients with mild to-moderate depression. Antidepressants are frequently used to treat irritable bowel syndrome (IBS). To date, no study that examines the efficacy of SJW in IBS has been carried out. The aim of this study was to evaluate the efficacy of SJW in IBS after 12 weeks. METHODS: In this randomized, double-blind, placebo controlled trial, 70 participants with an established diagnosis of IBS were randomized and assigned by concealed allocation to either SJW or placebo. Both treatment arms were balanced on symptom subtype. The primary end point was self reported overall bowel symptom score (BSS) at 12 weeks. Secondary end points were individual BSS for diarrhea (D-BSS), constipation (C-BSS), pain or discomfort, and bloating; adequate relief (AR) of IBS on at least 50% of the last 4 weeks of therapy; and IBS quality-of-life score at 12 weeks. RESULTS: In all, 86% of the participants were women, and the median age was 42 years. Overall, 29% had C-IBS, 37% D-IBS, and 31% had mixed IBS. Both groups reported decreases in overall BSS from baseline, with the placebo arm having significantly lower scores at 12 weeks (P=0.03) compared with SJW. These patterns of improvement were mirrored in the secondary end points with the placebo group faring better than the SJW-treated group, with significant differences observed at week 12 for D-BSS (P=0.03) and percent with AR (P=0.02). A similar proportion of subjects in each treatment group (SJW: 51% vs. placebo: 54%) believed that the study drug they received decreased IBS life interferences (P=0.79). CONCLUSIONS: SJW was a less effective treatment for IBS than placebo. PMID- 19809409 TI - Single-balloon enteroscopy-assisted ERCP in patients with Billroth II gastrectomy or Roux-en-Y anastomosis (with video). AB - OBJECTIVES: The aim of this study was to evaluate the usefulness of single balloon enteroscopy (SBE)-assisted endoscopic retrograde cholangiopancreatography (ERCP) in patients with Billroth II gastrectomy (B-II) or Roux-en-Y anastomosis (R-Y). METHODS: Thirteen SBE procedures were performed in 2 patients with B-II and 11 patients with R-Y at Tokyo Medical University Hospital. SBE was replaced with a conventional forward-viewing upper endoscope after reaching the papilla. This is a retrospective cohort study. RESULTS: In all cases but one (92.3%, 12/13), the papilla or anastomotic site could be reached with the SBE. The rate for B-II and R-Y were 100% (2/2) and 90.9% (10/11), respectively. The mean time required to reach the papilla or anastomotic site was 33.3 min (range, 5-86 min). In cases of R-Y, the mean time required to reach the papilla or anastomotic site was 35.5 min (range, 12-76 min). The overall success rate of the therapeutic ERCP on the first session was 76.9% (10/13). In patients with an intact papilla, the success rate on the first session was 72.3% (8/11). With regard to the type of surgery, the success rate of the procedure in patients with B-II and R-Y was 100% (2/2) and 72.3% (8/11), respectively. The mean procedural time for 10 successful groups on the first session was 66.4 min (range, 25-152 min). No adverse events occurred. CONCLUSIONS: SBE-assisted ERCP using an overtube-assisted technique appears to be promising for performing therapeutic ERCP in patients with B-II or R-Y. PMID- 19809410 TI - Association of higher DEFB4 genomic copy number with Crohn's disease. AB - OBJECTIVES: Human beta-defensin 2 (hBD-2 or DEFB4) is a highly inducible, antimicrobial peptide, which may have an important role in the innate immune response at epithelial surfaces. Genomic copy number of DEFB4 is polymorphic, with most individuals possessing 3-5 copies. Increased DEFB4 copy number is a susceptibility factor for psoriasis, whereas a single study in a Crohn's disease (CD) cohort reported that decreased DEFB4 copy number is associated with colonic inflammation. Here, we analyze association of DEFB4 copy number with CD in a New Zealand case-control cohort of European origin. METHODS: DEFB4 gene copy number was determined using TaqMan quantitative PCR in 466 CD patients and 329 controls. DNA samples, independently genotyped for DEFB4 copy number by alternative methods, were used to validate the assay. RESULTS: Increased DEFB4 genomic copy number was seen in CD patients compared with controls. Individuals with >4 copies had a significantly higher risk of developing CD than those with <4 copies (odds ratio 1.54; 95% confidence interval 1.13-2.09, P=5e-05). DEFB4 genomic copy number did not differ by disease location within the CD cohort (P=0.948), nor did analysis of CD patients who had undergone surgery detect association of decreased DEFB4 genomic copy number (<4) in colonic CD compared with ileal CD (P=0.120). CONCLUSIONS: Our results indicate that elevated DEFB4 copy number is a risk factor for CD (irrespective of intestinal location), and challenge previous data supporting positive association of lower DEFB4 genomic copy number with colonic CD. PMID- 19809411 TI - Interaction of adipokines and hepatitis B virus on histological liver injury in the Chinese. AB - OBJECTIVES: Chronic hepatitis B patients with diabetes and metabolic syndrome are at increased risk of cirrhosis and hepatocellular carcinoma, but the underlying mechanism is unclear. Our objective was to test whether dysregulation of adipokines contributes to liver injury. We also studied whether viral factors affected adipokines, insulin resistance, and hepatic steatosis. METHODS: A prospective cohort of 266 chronic hepatitis B patients undergoing liver biopsy was studied. Fasting blood was taken for the analysis of tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), adiponectin, leptin, and resistin. Insulin resistance was assessed by the homeostasis model assessment of insulin resistance (HOMA-IR). Factors associated with significant necroinflammation and cirrhosis were identified. RESULTS: Histological activity index was correlated with serum TNF-alpha (R=0.40, P<0.0001) and IL-6 (R=0.32, P<0.0001) but not with adiponectin, leptin, or resistin. By multivariate analysis, TNF-alpha was associated with significant necroinflammation after adjusting for age and viral factors (odds ratio (OR) 1.041, 95% confidence interval (CI) 1.002-1.082, P=0.04). Serum adiponectin had positive correlation with hepatitis B virus DNA (R=0.17, P=0.007) and was decreased in patients with insulin resistance and hepatic steatosis. On the other hand, viral load, hepatitis B e-antigen status, and genotypes had no association with insulin resistance, hepatic steatosis, and the levels of TNF-alpha and IL-6. A total of 68 (25.6%) patients had cirrhosis. HOMA-IR, but not adipokine dysregulation, was independently associated with cirrhosis (OR 1.09, 95% CI 1.02-1.15, P=0.006). CONCLUSIONS: TNF-alpha and/or IL 6 contribute to hepatic necroinflammation in chronic hepatitis B patients. Adiponectin protects against insulin resistance and hepatic steatosis but does not affect liver injury. Adipokines and viral factors contribute to liver injury independently. PMID- 19809412 TI - Efficacy of three different dosages of esomeprazole in the long-term management of reflux disease: a prospective, randomized study, using the wireless Bravo pH system. AB - OBJECTIVES: Gastroesophageal reflux disease (GERD) is a chronic condition that usually requires long-term maintenance therapy with proton-pump inhibitors (PPIs). In clinical practice, patients receive PPIs at the lowest dose to control symptoms. However, it is not known whether this approach adequately controls acidic esophageal reflux. We sought to investigate the efficacy of three different dosages of esomeprazole in patients receiving maintenance therapy for GERD, using the Bravo pH system. METHODS: Patients with a previous history of erosive esophagitis A or B (LA classification) that was healed at the time of enrollment or endoscopy-negative reflux disease (ENRD), documented with an abnormal pH study, were randomized to receive maintenance therapy with esomeprazole 40 mg twice daily (group A), once daily (group B), or every other day (group C). Intraesophageal pH was monitored for two consecutive days using the Bravo wireless system, 30 days after randomization. The parameters subjected to analysis were percent of total time pH<4 and the De Meester score. RESULTS: The pH results from 73 patients (group A=24, group B=24, group C=25 patients) were subjected to final analysis. On the first day of the study, the mean (+/ s.d.) percent of total time pH <4 and the De Meester score were group A: 0.9(1.2) and 4.1(4.0); group B: 1.5(1.6) and 7.0(6.9); group C: 1.3(1.0) and 6.0(3.3), respectively (P=0.262 and 0.134, respectively). On the second day of the study, the corresponding values were group A: 0.7(1.0) and 3.9(5.9); group B: 1.5(1.8) and 6.4(6.6); group C: 7.0(4.4) and 29.4(19.4), respectively. The difference was statistically significant (P<0.0001 and <0.0001, respectively). Further analysis showed that patients not receiving PPI had a significantly higher mean percent of total time pH<4 and De Meester score as compared with patients on PPI once or twice daily (P<0.001 and <0.001 respectively). CONCLUSIONS: The administration of esomeprazole 40 mg every other day does not control acidic esophageal reflux on the day off PPI. Esomeprazole 40 mg once daily effectively controls reflux of acid in patients with history of mild esophagitis or ENRD, whereas doubling the dose does not seem to confer any further advantage. PMID- 19809413 TI - Long-term use of statins and risk of colorectal cancer: a population-based study. AB - OBJECTIVES: We conducted a population-based cohort study to determine the effect of long-term regular use of statins on the risk of colorectal cancer (CRC). METHODS: Individuals who were dispensed statins regularly were identified from Manitoba's population-based prescription drug database and followed up until diagnosis of CRC, migration out of province, death, or December 2005. The incidence of CRC in this group was compared with that among individuals who were never dispensed statins. Stratified analysis was performed to determine the risk after 5 years of regular statin use. Multivariate Poisson regression models were used to adjust for potential confounding by age, sex, and history of diabetes, inflammatory bowel disease, coronary heart disease, lower gastrointestinal endoscopy, resective colorectal surgery, use of nonsteroidal anti-inflammatory drugs, hormone replacement therapy (among women), and median household income. The dose effect was evaluated in defined daily dose units. RESULTS: In total, 35,739 individuals were dispensed statins regularly. In all, 10,287 (49% males; 51% females) long-term (>or=5 years) regular statin users were followed up for up to 5 additional years. In multivariate analysis, the incidence rate ratio (IRR) of CRC among those dispensed statins regularly compared with those who were never dispensed statins (n=377,532) was 1.13 (95% confidence interval (CI): 1.02-1.25). The CRC risk among the long-term regular statin users was similar to that for individuals never dispensed statins (IRR, 0.89; 95% CI: 0.70-1.13). A statistically nonsignificant risk reduction was observed among high-dose long term regular statin users. CONCLUSIONS: These findings suggest that long-term regular use of statins for the current clinical indications does not protect against CRC. The benefit of high-dose long-term statin use needs further evaluation. PMID- 19809414 TI - Trends in ambulatory and emergency room visits for inflammatory bowel diseases in the United States: 1994-2005. AB - OBJECTIVES: Inflammatory bowel diseases (IBDs) require frequent healthcare encounters. Analysis of trends in disease burden is important to study the changing epidemiology of disease. The aim of our study was to examine national trends in IBD-related visits in ambulatory and emergency room (ER) settings. METHODS: We used data from the National Ambulatory Medical Care Survey (NAMCS) and National Hospital Ambulatory Medical Care Survey (NHAMCS) using the International Classification of Diseases, 9th Revision, Clinical Modification codes to identify IBD-related visits. The annualized number of visits was examined in 3-year time periods from 1994 to 2005. We compared the demographic distribution of the inpatient, outpatient, and ER IBD cohorts. RESULTS: There were an estimated 1.1 million (95% confidence interval (CI): 0.9-1.4 million) annual IBD-related visits during the period 1994-1996 increasing to 1.8 million (95% CI: 1.4-2.2 million) visits during 2003-2005 (+55%). Correspondingly, there were 28,752 estimated annual ER visits (95% CI: 16,299-41,206) related to IBD during 1994-1996 increasing to 76,374 annual visits (95% CI: 42,498-112,257) during 2003-2005 (+165%). The proportion of outpatient visits by patients older than 50 years increased from 34.1 to 52.0% (P<0.0001). Use of corticosteroids decreased from 26.5% of visits to 14.5% (P<0.0001), with a corresponding rise in immunomodulator/biologic use (2.6-13.8%, P<0.0001). Younger, self-pay patients and those with Crohn's disease were over-represented in the ER IBD cohort compared with inpatients or outpatients. CONCLUSIONS: There has been an increase in IBD-related ambulatory and ER visits from 1994 to 2005. There is a need for research examining the reasons for the disproportionate representation of certain demographic groups in the ER setting. PMID- 19809415 TI - Ulcerative colitis and pregnancy outcomes in an Asian population. AB - OBJECTIVES: As the prevalence of ulcerative colitis (UC) is much higher in Western countries than in Asian countries, previous investigations of pregnancy outcomes for women with UC were limited to people of European descent. This study was aimed at examining the risk of adverse pregnancy outcomes (low birth weight (LBW), preterm birth, small for gestational age (SGA), and cesarean section (CS)) among Asian women with UC. METHODS: Using a 3-year nationwide population-based database, we identified a total of 196 women who gave birth from 2001 to 2003, who were diagnosed with UC within 2 years before their index deliveries. A total of 1,568 unaffected pregnant women matched these cases according to age and year of delivery. Conditional logistic regression analyses were performed to estimate risk. RESULTS: There were significant differences between women with and without UC in terms of LBW (12.76% vs. 5.55, P<0.001) and preterm births (11.73% vs. 6.25%, P=0.004). After adjusting for infant gender, parity, maternal age, highest maternal educational level, parental age difference, maternal marital status, hypertension, diabetes, anemia, family monthly income, as well as conditioning on maternal age and year of delivery, the odds of LBW and preterm births for women with UC were 2.36 (95% confidence interval (CI)=1.45-3.82) and 1.90 (95% CI=1.16 3.11) times, respectively, those for unaffected women. CONCLUSIONS: Although UC often follows a milder disease course in Asians than in people of European descent, we demonstrated that Asian women suffering from UC were still at risk of having preterm and LBW babies, compared with unaffected mothers. PMID- 19809419 TI - Vascular endothelial growth factor delivery via gene therapy for diabetic wounds: first steps. PMID- 19809420 TI - Achieving successful delivery of nucleic acids to skin: 6th Annual Meeting of the International Pachyonychia Congenita Consortium. PMID- 19809421 TI - The central role of Bcl-3 in atopic dermatitis. AB - Cytokines and hormones are essential mediators in the regulation of antimicrobial peptide and protein (AMP) levels during host defense responses. In this issue, Buchau et al. identify and characterize B-cell leukemia-3 as an important modulator of AMP expression during the innate immune response in keratinocytes as well as in the chronic inflammatory skin disorder atopic dermatitis. PMID- 19809422 TI - Antimicrobial RNases of human skin. AB - Included in the armamentarium of antimicrobial defenses of human skin are the RNases. In this issue, Abtin et al. report new details about how this antimicrobial system is deployed. Although present throughout the epidermis, RNases appear to be free to act only within the stratum corneum. Elsewhere in the epidermis RNases are complexed with an inhibitory protein. In the stratum corneum proteases degrade the inhibitor, freeing the RNase and liberating it to function in antimicrobial defense. PMID- 19809423 TI - Propionibacterium acnes and sebaceous lipogenesis: a love-hate relationship? AB - In this issue, Iinuma et al. show that Propionibacterium acnes (P. acnes) conditioned medium and formalin-killed P. acnes augment intracellular lipid formation in hamster sebocytes by increasing the de novo synthesis of triacylglycerols. This commentary summarizes the current knowledge of the association of P. acnes with sebaceous lipogenesis, inflammation, and innate immunity, and points out the concurrent evidence that P. acnes-induced lipids may represent a recruitment of allies and/or enemies of the human skin. PMID- 19809424 TI - An "ice-cold" TR(i)P to skin biology: the role of TRPA1 in human epidermal keratinocytes. AB - Recent studies have suggested the expression of numerous heat-sensitive transient receptor potential (TRP) ion channels in non-neuronal cell populations of the skin. In this issue, Atoyan et al. provide evidence that the noxious cold activated TRPA1 is widely expressed in various human cutaneous cells and that it may be directly involved in the regulation of keratinocyte proliferation and differentiation and in cutaneous inflammatory responses. PMID- 19809425 TI - Tumorigenic effect of moisturizing creams in UVB-pretreated high-risk mice. PMID- 19809427 TI - Gene expression profiling associated with the progression to poorly differentiated thyroid carcinomas. AB - BACKGROUND: Poorly differentiated thyroid carcinomas (PDTC) represent a heterogeneous, aggressive entity, presenting features that suggest a progression from well-differentiated carcinomas. To elucidate the mechanisms underlying such progression and identify novel therapeutic targets, we assessed the genome-wide expression in normal and tumour thyroid tissues. METHODS: Microarray analyses of 24 thyroid carcinomas - 7 classic papillary, 8 follicular variants of papillary (fvPTC), 4 follicular (FTC) and 5 PDTC - were performed and correlated with RAS, BRAF, RET/PTC and PAX8-PPARG alterations. Selected genes were validated by quantitative RT-PCR in an independent set of 28 thyroid tumours. RESULTS: Unsupervised analyses showed that gene expression similarity was higher between PDTC and fvPTC, particularly for tumours harbouring RAS mutations. Poorly differentiated thyroid carcinomas presented molecular signatures related to cell proliferation, poor prognosis, spindle assembly checkpoint and cell adhesion. Compared with normal tissues, PTC had 307 out of 494 (60%) genes over-expressed, FTC had 137 out of 171 (80%) genes under-expressed, whereas PDTC had 92 out of 107 (86%) genes under-expressed, suggesting that gene downregulation is involved in tumour dedifferentiation. Significant UHRF1 and ITIH5 deregulated gene expression in PDTC, relatively to normal tissues, was confirmed by quantitative RT-PCR. CONCLUSION: Our findings suggest that fvPTC are possible precursors of PDTC. Furthermore, UHRF1 and ITIH5 have a potential therapeutic/prognostic value for aggressive thyroid tumours. PMID- 19809428 TI - Inhibition of constitutive and cxc-chemokine-induced NF-kappaB activity potentiates ansamycin-based HSP90-inhibitor cytotoxicity in castrate-resistant prostate cancer cells. AB - BACKGROUND: We determined how CXC-chemokine signalling and necrosis factor-kappaB (NF-kappaB) activity affected heat-shock protein 90 (Hsp90) inhibitor (geldanamycin (GA) and 17-allylamino-demethoxygeldanamycin (17-AAG)) cytotoxicity in castrate-resistant prostate cancer (CRPC). METHODS: Geldanamycin and 17-AAG toxicity, together with the CXCR2 antagonist AZ10397767 or NF-kappaB inhibitor BAY11-7082, was assessed by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide assay in two CRPC lines, DU145 and PC3. Flow cytometry quantified apoptotic or necrosis profiles. Necrosis factor-kappaB activity was determined by luciferase readouts or indirectly by quantitative PCR and ELISA-based determination of CXCL8 expression. RESULTS: Geldanamycin and 17 AAG reduced PC3 and DU145 cell viability, although PC3 cells were less sensitive. Addition of AZ10397767 increased GA (e.g., PC3 IC(20): from 1.67+/-0.4 to 0.18+/ 0.2 nM) and 17-AAG (PC3 IC(20): 43.7+/-7.8 to 0.64+/-1.8 nM) potency in PC3 but not DU145 cells. Similarly, BAY11-7082 increased the potency of 17-AAG in PC3 but not in DU145 cells, correlating with the elevated constitutive NF-kappaB activity in PC3 cells. AZ10397767 increased 17-AAG-induced apoptosis and necrosis and decreased NF-kappaB activity/CXCL8 expression in 17-AAG-treated PC3 cells. CONCLUSION: Ansamycin cytotoxicity is enhanced by inhibiting NF-kappaB activity and/or CXC-chemokine signalling in CRPC cells. Detecting and/or inhibiting NF kappaB activity may aid the selection and treatment response of CRPC patients to Hsp90 inhibitors. PMID- 19809429 TI - Change in the hormone receptor status following administration of neoadjuvant chemotherapy and its impact on the long-term outcome in patients with primary breast cancer. AB - BACKGROUND: To evaluate the impact of change in the hormone receptor (HR) status (HR status conversion) on the long-term outcomes of breast cancer patients treated with neoadjuvant chemotherapy (NAC). METHODS: We investigated 368 patients for the HR status of their lesions before and after NAC. On the basis of the HR status and the use/non-use of endocrine therapy (ET), the patients were categorised into four groups: Group A, 184 ET-administered patients with HR positive both before and after NAC; Group B, 47 ET-administered patients with HR status conversion; Group C, 12 ET-naive patients with HR status conversion; Group D, 125 patients with HR-negative both before and after NAC. RESULTS: Disease-free survival in Group B was similar to that in Group A (hazard ratio, 1.16; P=0.652), but that in Group C was significantly lesser than that in Group A (hazard ratio, 6.88; P<0.001). A similar pattern of results was obtained for overall survival. CONCLUSION: Our results indicate that the HR status of tumours is a predictive factor for disease-free and overall survival and that ET appears to be suitable for patients with HR status conversion. Therefore, both the CNB and surgical specimens should be monitored for HR status. PMID- 19809431 TI - A survey of adolescent experiences of human papillomavirus vaccination in the Manchester study. AB - BACKGROUND: There is little information on girls' experiences of human papillomavirus (HPV) vaccination in the prevention of cervical cancer. We investigated the views of adolescent girls who had been offered the vaccine as part of a feasibility study conducted in Manchester. METHODS: All 12 to 13-year old girls in two primary care trusts were offered three doses of Cervarix (manufactured by GlaxoSmithKline). A letter was sent to 1084 parents who had consented to research follow-up. It requested parents to pass a questionnaire regarding HPV vaccination to their daughters to complete and post back in a prepaid envelope. RESULTS: A total of 553 girls completed the questionnaire. Altogether, 77% (422) had shared with their parents in the vaccine decision. In all, 42% (n=13) of girls, whose parents refused vaccination, stated that they wanted the vaccine, whereas 10% (50) of those who were vaccinated did not want the vaccine. Although 54% (277) said the vaccine was very important to them, 39% (153) of vaccinated girls thought they might not recommend it to others. The vaccine was perceived to be painful and there were exaggerated rumours of serious adverse events and needle scares. A total of 79% (420) of girls agreed with a statement that vaccination reminded them of the risks of sexual contact, but 14% (73) agreed they might take more sexual risks because they had been vaccinated. CONCLUSION: Girls of this age form their own views on HPV vaccination but parental support for vaccination remains important, especially for completing the three doses. By discussing the vaccine, parents can encourage their daughters to determine the importance and implications of HPV vaccination. PMID- 19809432 TI - Clinical application of fluorescence endoscopic imaging using hypericin for the diagnosis of human oral cavity lesions. AB - BACKGROUND: Diagnosis of oral cancer is conventionally carried out using white light endoscopy and histopathology of biopsy samples. However, oral tumours are mostly superficial and the lesion and its margins can be difficult to visualise under white light. We present clinical data on fluorescence diagnostic imaging of oral lesions using hypericin, a plant-based photosensitiser. METHODS: Fluorescence images of lesions and normal tissue were captured using an endoscope after hypericin administration. The images were analysed to extract their colour parameters, which, along with the red-to-blue intensity ratios, were analysed and used to discriminate between tissue types. The results were correlated with those from histopathology. RESULTS: The red-to-blue intensity ratio increased from normal to hyperplastic to cancerous tissue and was a good parameter to discriminate between these tissue types, with sensitivity and specificity levels of 90% and above. CONCLUSION: Our results show that hypericin fluorescence imaging has the potential to be used for the clinical diagnosis of oral cancer. Further study to enhance the clinical potential of this technique includes the development of a real-time image processing and analysis system interfaced to the endoscope to enable same-day cancer diagnosis and demarcation of lesion margins in a clinical setting. PMID- 19809433 TI - The state of the art: immune-mediated mechanisms of monoclonal antibodies in cancer therapy. AB - A number of antibody products have now become accepted as effective anti-cancer therapies. Despite being mainly designed to act by inhibiting functional tumour antigens, there is increasing evidence that Fc-mediated engagement of the immune system is an important contributor to the efficacy of several of these therapies. The optimisation of this engagement offers the potential not only to augment efficacy against existing targets, but also to exploit non-functional tumour antigens. Antibodies that achieve efficacy wholly or predominantly through Fc mediated mechanisms, represent rich opportunities for future therapeutics in oncology. This mini review summarises some of the key challenges, which need to be addressed to select the most effective molecules. These include the identification of optimal antibody characteristics and improvement of the drug discovery process, in particular, the relevance and predictive power of existing in vitro and in vivo screening methods. Advances in our understanding of tumour immunobiology and successful application of technologies designed to enhance immune system engagement will further aid this process. PMID- 19809434 TI - Glasgow Prognostic Score as a predictive factor differentiating surgical site infection and remote infection following colorectal cancer surgery? PMID- 19809435 TI - Curcumin induces apoptosis-independent death in oesophageal cancer cells. AB - BACKGROUND: Oesophageal cancer incidence is increasing and survival rates remain extremely poor. Natural agents with potential for chemoprevention include the phytochemical curcumin (diferuloylmethane). We have examined the effects of curcumin on a panel of oesophageal cancer cell lines. METHODS: MTT (3-(4,5 dimethyldiazol-2-yl)-2,5 diphenyl tetrazolium bromide) assays and propidium iodide staining were used to assess viability and DNA content, respectively. Mitotic catastrophe (MC), apoptosis and autophagy were defined by both morphological criteria and markers such as MPM-2, caspase 3 cleavage and monodansylcadaverine (MDC) staining. Cyclin B and poly-ubiquitinated proteins were assessed by western blotting. RESULTS: Curcumin treatment reduces viability of all cell lines within 24 h of treatment in a 5-50 muM range. Cytotoxicity is associated with accumulation in G2/M cell-cycle phases and distinct chromatin morphology, consistent with MC. Caspase-3 activation was detected in two out of four cell lines, but was a minor event. The addition of a caspase inhibitor zVAD had a marginal or no effect on cell viability, indicating predominance of a non apoptotic form of cell death. In two cell lines, features of both MC and autophagy were apparent. Curcumin-responsive cells were found to accumulate poly ubiquitinated proteins and cyclin B, consistent with a disturbance of the ubiquitin-proteasome system. This effect on a key cell-cycle checkpoint regulator may be responsible for the mitotic disturbances and consequent cytotoxicity of this drug. CONCLUSION: Curcumin can induce cell death by a mechanism that is not reliant on apoptosis induction, and thus represents a promising anticancer agent for prevention and treatment of oesophageal cancer. PMID- 19809439 TI - Organics settle down. PMID- 19809436 TI - A phase-II trial of dose-dense chemotherapy in patients with disseminated thymoma: report of a Japan Clinical Oncology Group trial (JCOG 9605). AB - BACKGROUND: To evaluate the safety and efficacy of dose-dense weekly chemotherapy in the treatment of advanced thymoma. METHODS: Subjects comprised patients with histologically documented chemotherapy-naive thymoma with stage-IVa or IVb disease. Thymic carcinoma, carcinoid or lymphoma cases were excluded. Patients received 9 weeks of chemotherapy: cisplatin (25 mg m(-2)) on weeks 1-9; vincristine (1 mg m(-2)) on weeks 1, 2, 4, 6 and 8; and doxorubicin (40 mg m(-2)) and etoposide (80 mg m(-2)) on days 1-3 of weeks 1, 3, 5, 7 and 9. Chemotherapy courses were supported by granulocyte colony-stimulating factor. Post-protocol local therapy was allowed. RESULTS: From July 1997 to March 2004, 30 patients were entered. Three were ineligible due to different histology. Chemotherapy associated toxicity was mainly haematological and was well tolerated, with no deaths due to toxicity, and 87% of patients completed the planned 9-week regimen. Overall response rate was 59%, with 16 of the 27 eligible patients achieving partial response. Median progression-fee survival (PFS) was 0.79 years (95% confidence interval: 0.52-1.40 years), and PFS at 1 and 2 years was 37 and 15%, respectively. Overall survival rates at 2 and 5 years were 89 and 65%, respectively. CONCLUSION: In stage-IV thymoma patients, weekly dose-dense chemotherapy offers similar activity to conventional regimens. PMID- 19809440 TI - The new deficit model. PMID- 19809441 TI - Selling graphene by the ton. PMID- 19809442 TI - It's not just about nanotoxicology. PMID- 19809445 TI - Nanoelectromechanical systems: Show of strength. PMID- 19809446 TI - Probe microscopy: A closer look at the atoms in a molecule. PMID- 19809447 TI - Nanomedicine: Sniffing out lung cancer. PMID- 19809448 TI - Nanopatterning: Surfaces feel the heat. PMID- 19809449 TI - Spintronics: Shedding light on nanomagnets. PMID- 19809450 TI - Biomolecular computing: Molecules that reason. PMID- 19809452 TI - Promises, facts and challenges for carbon nanotubes in imaging and therapeutics. AB - The use of carbon nanotubes in medicine is now at the crossroads between a proof of-principle concept and an established preclinical candidate for a variety of therapeutic and diagnostic applications. Progress towards clinical trials will depend on the outcomes of efficacy and toxicology studies, which will provide the necessary risk-to-benefit assessments for carbon-nanotube-based materials. Here we focus on carbon nanotubes that have been studied in preclinical animal models, and draw attention to the promises, facts and challenges of these materials as they transition from research to the clinical phase. We address common questions regarding the use of carbon nanotubes in disease imaging and therapy, and highlight the opportunities and challenges ahead. PMID- 19809453 TI - Towards a definition of inorganic nanoparticles from an environmental, health and safety perspective. AB - The regulation of engineered nanoparticles requires a widely agreed definition of such particles. Nanoparticles are routinely defined as particles with sizes between about 1 and 100 nm that show properties that are not found in bulk samples of the same material. Here we argue that evidence for novel size dependent properties alone, rather than particle size, should be the primary criterion in any definition of nanoparticles when making decisions about their regulation for environmental, health and safety reasons. We review the size dependent properties of a variety of inorganic nanoparticles and find that particles larger than about 30 nm do not in general show properties that would require regulatory scrutiny beyond that required for their bulk counterparts. PMID- 19809454 TI - Molecular implementation of simple logic programs. AB - Autonomous programmable computing devices made of biomolecules could interact with a biological environment and be used in future biological and medical applications. Biomolecular implementations of finite automata and logic gates have already been developed. Here, we report an autonomous programmable molecular system based on the manipulation of DNA strands that is capable of performing simple logical deductions. Using molecular representations of facts such as Man(Socrates) and rules such as Mortal(X) <-- Man(X) (Every Man is Mortal), the system can answer molecular queries such as Mortal(Socrates)? (Is Socrates Mortal?) and Mortal(X)? (Who is Mortal?). This biomolecular computing system compares favourably with previous approaches in terms of expressive power, performance and precision. A compiler translates facts, rules and queries into their molecular representations and subsequently operates a robotic system that assembles the logical deductions and delivers the result. This prototype is the first simple programming language with a molecular-scale implementation. PMID- 19809455 TI - Single-crystal germanium layers grown on silicon by nanowire seeding. AB - Three-dimensional integration and the combination of different material systems are central themes of electronics research. Recently, as-grown vertical one dimensional structures have been integrated into high-density three-dimensional circuits. However, little attention has been paid to the unique structural properties of germanium nanowires obtained by epitaxial and heteroepitaxial growth on Ge(111) and Si(111) substrates, despite the fact that the integration of germanium on silicon is attractive for device applications. Here, we demonstrate the lateral growth of single crystal germanium islands tens of micrometres in diameter by seeding from germanium nanowires grown on a silicon substrate. Vertically aligned high-aspect-ratio nanowires can transfer the orientation and perfection of the substrate crystal to overlying layers a micrometre or more above the substrate surface. This technique can be repeated to build multiple active device layers, a key requirement for the fabrication of densely interconnected three-dimensional integrated circuits. PMID- 19809456 TI - Catalyst preparation for CMOS-compatible silicon nanowire synthesis. AB - Metallic contamination was key to the discovery of semiconductor nanowires, but today it stands in the way of their adoption by the semiconductor industry. This is because many of the metallic catalysts required for nanowire growth are not compatible with standard CMOS (complementary metal oxide semiconductor) fabrication processes. Nanowire synthesis with those metals that are CMOS compatible, such as aluminium and copper, necessitate temperatures higher than 450 degrees C, which is the maximum temperature allowed in CMOS processing. Here, we demonstrate that the synthesis temperature of silicon nanowires using copper based catalysts is limited by catalyst preparation. We show that the appropriate catalyst can be produced by chemical means at temperatures as low as 400 degrees C. This is achieved by oxidizing the catalyst precursor, contradicting the accepted wisdom that oxygen prevents metal-catalysed nanowire growth. By simultaneously solving material compatibility and temperature issues, this catalyst synthesis could represent an important step towards real-world applications of semiconductor nanowires. PMID- 19809457 TI - Uniform exciton fluorescence from individual molecular nanotubes immobilized on solid substrates. AB - Self-assembled quasi one-dimensional nanostructures of pi-conjugated molecules may find a use in devices owing to their intriguing optoelectronic properties, which include sharp exciton transitions, strong circular dichroism, high exciton mobilities and photoconductivity. However, many applications require immobilization of these nanostructures on a solid substrate, which is a challenge to achieve without destroying their delicate supramolecular structure. Here, we use a drop-flow technique to immobilize double-walled tubular J-aggregates of amphiphilic cyanine dyes without affecting their morphological or optical properties. High-resolution images of the topography and exciton fluorescence of individual J-aggregates are obtained simultaneously with polarization-resolved near-field scanning optical microscopy. These images show remarkably uniform supramolecular structure, both along individual nanotubes and between nanotubes in an ensemble, demonstrating their potential for light harvesting and energy transport. PMID- 19809458 TI - Thermochemical nanopatterning of organic semiconductors. AB - Patterning of semiconducting polymers on surfaces is important for various applications in nanoelectronics and nanophotonics. However, many of the approaches to nanolithography that are used to pattern inorganic materials are too harsh for organic semiconductors, so research has focused on optical patterning and various soft lithographies. Surprisingly little attention has been paid to thermal, thermomechanical and thermochemical patterning. Here, we demonstrate thermochemical nanopatterning of poly(p-phenylene vinylene), a widely used electroluminescent polymer, by a scanning probe. We produce patterned structures with dimensions below 28 nm, although the tip of the probe has a diameter of 5 microm, and achieve write speeds of 100 microm s(-1). Experiments show that a resolution of 28 nm is possible when the tip-sample contact region has dimensions of approximately 100 nm and, on the basis of finite-element modelling, we predict that the resolution could be improved by using a thinner resist layer and an optimized probe. Thermochemical lithography offers a versatile, reliable and general nanopatterning technique because a large number of optical materials, including many commercial crosslinker additives and photoresists, rely on chemical mechanisms that can also be thermally activated. PMID- 19809459 TI - Diagnosing lung cancer in exhaled breath using gold nanoparticles. AB - Conventional diagnostic methods for lung cancer are unsuitable for widespread screening because they are expensive and occasionally miss tumours. Gas chromatography/mass spectrometry studies have shown that several volatile organic compounds, which normally appear at levels of 1-20 ppb in healthy human breath, are elevated to levels between 10 and 100 ppb in lung cancer patients. Here we show that an array of sensors based on gold nanoparticles can rapidly distinguish the breath of lung cancer patients from the breath of healthy individuals in an atmosphere of high humidity. In combination with solid-phase microextraction, gas chromatography/mass spectrometry was used to identify 42 volatile organic compounds that represent lung cancer biomarkers. Four of these were used to train and optimize the sensors, demonstrating good agreement between patient and simulated breath samples. Our results show that sensors based on gold nanoparticles could form the basis of an inexpensive and non-invasive diagnostic tool for lung cancer. PMID- 19809460 TI - Monolayer coverage and channel length set the mobility in self-assembled monolayer field-effect transistors. AB - The mobility of self-assembled monolayer field-effect transistors (SAMFETs) traditionally decreases dramatically with increasing channel length. Recently, however, SAMFETs using liquid-crystalline molecules have been shown to have bulk like mobilities that are virtually independent of channel length. Here, we reconcile these scaling relations by showing that the mobility in liquid crystalline SAMFETs depends exponentially on the channel length only when the monolayer is incomplete. We explain this dependence both numerically and analytically, and show that charge transport is not affected by carrier injection, grain boundaries or conducting island size. At partial coverage, that is when the monolayer is incomplete, liquid-crystalline SAMFETs thus form a unique model system to study size-dependent conductance originating from charge percolation in two dimensions. PMID- 19809461 TI - Charge-controlled magnetism in colloidal doped semiconductor nanocrystals. AB - Electrical control over the magnetic states of doped semiconductor nanostructures could enable new spin-based information processing technologies. To this end, extensive research has recently been devoted to examination of carrier-mediated magnetic ordering effects in substrate-supported quantum dots at cryogenic temperatures, with carriers introduced transiently by photon absorption. The relatively weak interactions found between dopants and charge carriers have suggested that gated magnetism in quantum dots will be limited to cryogenic temperatures. Here, we report the observation of a large, reversible, room temperature magnetic response to charge state in free-standing colloidal ZnO nanocrystals doped with Mn(2+) ions. Injected electrons activate new ferromagnetic Mn(2+)-Mn(2+) interactions that are strong enough to overcome antiferromagnetic coupling between nearest-neighbour dopants, making the full magnetic moments of all dopants observable. Analysis shows that this large effect occurs in spite of small pairwise electron-Mn(2+) exchange energies, because of competing electron-mediated ferromagnetic interactions involving distant Mn(2+) ions in the same nanocrystal. PMID- 19809462 TI - Golden carbon nanotubes as multimodal photoacoustic and photothermal high contrast molecular agents. AB - Carbon nanotubes have shown promise as contrast agents for photoacoustic and photothermal imaging of tumours and infections because they offer high resolution and allow deep tissue imaging. However, in vivo applications have been limited by the relatively low absorption displayed by nanotubes at near-infrared wavelengths and concerns over toxicity. Here, we show that gold-plated carbon nanotubes termed golden carbon nanotubes-can be used as photoacoustic and photothermal contrast agents with enhanced near-infrared contrast ( approximately 10(2)-fold) for targeting lymphatic vessels in mice using extremely low laser fluence levels of a few mJ cm(-2). Antibody-conjugated golden carbon nanotubes were used to map the lymphatic endothelial receptor, and preliminary in vitro viability tests show golden carbon nanotubes have minimal toxicity. This new nanomaterial could be an effective alternative to existing nanoparticles and fluorescent labels for non invasive targeted imaging of molecular structures in vivo. PMID- 19809463 TI - Cilostazol, a phosphodiesterase inhibitor, attenuates photothrombotic focal ischemic brain injury in hypertensive rats. AB - The aim of this study was to evaluate and compare the effects of anti-platelet agents with different modes of action (cilostazol, aspirin, and clopidogrel) on brain infarction produced by photothrombotic middle-cerebral-artery (MCA) occlusion in male, spontaneously hypertensive rats. Cerebral blood flow (CBF) was measured with laser-Doppler flowmetry in the penumbral cortex. Infarct size was evaluated 24 h after MCA occlusion. The effects of these drugs on infarct size were examined by pretreatment of rats undergoing MCA occlusion. Pretreatment with cilostazol (100 mg/kg) significantly reduced infarct size. In contrast, aspirin (10 mg/kg) and clopidogrel (3 mg/kg) failed to mitigate infarct size, regardless of their apparent inhibitory effects on platelet aggregation. Post-treatment with cilostazol also significantly attenuated the infarct size, associated with improved CBF in the penumbral region. In support of this effect, cilostazol increased nitric oxide (NO) production and prostaglandin-I(2) (PGI(2)) release in cultured human brain microvascular endothelial cells. Cilostazol-induced NO production and PGI(2) release were completely abolished by an NO synthase inhibitor and aspirin, respectively. These findings show that cilostazol reduced brain infarct size due to an improvement in penumbral CBF possibly in association with increased endothelial NO and PGI(2) production. PMID- 19809464 TI - Arterial spin labeling perfusion MRI at multiple delay times: a correlative study with H(2)(15)O positron emission tomography in patients with symptomatic carotid artery occlusion. AB - Arterial spin labeling (ASL) perfusion magnetic resonance imaging (MRI) with image acquisition at multiple inversion times is a noninvasive ASL technique able to compensate for spatial heterogeneities in transit times caused by collateral blood flow in patients with severe stenosis of the cerebropetal blood vessels. Our aim was to compare ASL-MRI and H(2)(15)O positron emission tomography (PET), the gold standard for cerebral blood flow (CBF) assessment, in patients with a symptomatic internal carotid artery (ICA) occlusion. Fourteen patients (63+/-14 years) with a symptomatic ICA occlusion underwent both ASL-MRI and H(2)(15)O PET. The ASL-MRI was performed using a pulsed STAR labeling technique at multiple inversion times within 7 days of the PET. The CBF was measured in the gray-matter of the anterior, middle and posterior cerebral artery, and white-matter. Both PET and ASL-MRI showed a significantly decreased CBF in the gray-matter of the middle cerebral artery in the hemisphere ipsilateral to the ICA occlusion. The average gray-matter CBF measured with ASL-MRI (71.8+/-4.3 mL/min/100 g) was higher (P<0.01) than measured with H(2)(15)O PET (43.1+/-1.0 mL/min/100 g). In conclusion, ASL-MRI at multiple TIs is capable of depicting areas of regions with low CBF in patients with an occlusion of the ICA, although a systematic overestimation of CBF relative to H(2)(15)O PET was noted. PMID- 19809465 TI - The impact of erythropoietin on short-term changes in phosphorylation of brain protein kinases in a rat model of traumatic brain injury. AB - We found that recombinant human erythropoietin (rhEPO) reduced significantly the development of brain edema in a rat model of diffuse traumatic brain injury (TBI) (impact-acceleration model). In this study, we investigated the molecular and intracellular changes potentially involved in these immediate effects. Brain tissue nitric oxide (NO) synthesis, phosphorylation level of two protein kinases (extracellular-regulated kinase (ERK)-1/-2 and Akt), and brain water content were measured 1 (H1) and 2 h (H2) after insult. Posttraumatic administration of rhEPO (5,000 IU/kg body weight, intravenously, 30 mins after injury) reduced TBI induced upregulation of ERK phosphorylation, although it increased Akt phosphorylation at H1. These early molecular changes were associated with a reduction in brain NO synthesis at H1 and with an attenuation of brain edema at H2. Intraventricular administration of the ERK-1/-2 inhibitor, U0126, or the Akt inhibitor, LY294002, before injury showed that ERK was required for brain edema formation, and that rhEPO-induced reduction of edema could involve the ERK pathway. These results were obtained in the absence of any evidence of blood brain barrier damage on contrast-enhanced magnetic resonance images. The findings of our study indicate that the anti edematous effect of rhEPO could be mediated through an early inhibition of ERK phosphorylation after diffuse TBI. PMID- 19809466 TI - Glycogen metabolism as a marker of astrocyte differentiation. AB - Glycogen is a hallmark of mature astrocytes, but its emergence during astrocytic differentiation is unclear. Differentiation of E14 mouse neurospheres into astrocytes was induced with fetal bovine serum (FBS), Leukemia Inhibitory Factor (LIF), or Ciliary Neurotrophic Factor (CNTF). Cytochemical and enzymatic analyses showed that glycogen is present in FBS- or LIF- but not in CNTF-differentiated astrocytes. Glycogenolysis was induced in FBS- and LIF-differentiated astrocytes but glycogen resynthesis was observed only with FBS. Protein targeting to glycogen mRNA expression appeared with glial fibrillary acidic protein and S100beta in FBS and LIF conditions but not with CNTF. These results show that glycogen metabolism constitutes a useful marker of astrocyte differentiation. PMID- 19809468 TI - Acetazolamide-induced vasodilation does not inhibit the visually evoked flow response. AB - Different methods are used to assess the vasodilator ability of cerebral blood vessels; however, the exact mechanism of cerebral vasodilation, induced by different stimuli, is not entirely known. Our aim was to investigate whether the potent vasodilator agent, acetazolamide (AZ), inhibits the neurovascular coupling, which also requires vasodilation. Therefore, visually evoked flow parameters were examined by transcranial Doppler in ten healthy subjects before and after AZ administration. Pulsatility index and peak systolic flow velocity changes, evoked by visual stimulus, were recorded in the posterior cerebral arteries before and after intravenous administration of 15 mg/kg AZ. Repeated measures ANOVA did not show significant group main effect between the visually evoked relative flow velocity time courses before and after AZ provocation (P=0.43). Visual stimulation induced significant increase of relative flow velocity and decrease of pulsatility index not only before but also at the maximal effect of AZ. These results suggest that maximal cerebral vasodilation cannot be determined by the clinically accepted dose of AZ (15 mg/kg) and prove that neurovascular coupling remains preserved despite AZ-induced vasodilation. Our observation indicates independent regulation of vasodilation during neurovascular coupling, allowing the adaptation of cerebral blood flow according to neuronal activity even if other processes require significant vasodilation. PMID- 19809467 TI - Traumatic brain injury: an overview of pathobiology with emphasis on military populations. AB - This review considers the pathobiology of non-impact blast-induced neurotrauma (BINT). The pathobiology of traumatic brain injury (TBI) has been historically studied in experimental models mimicking features seen in the civilian population. These brain injuries are characterized by primary damage to both gray and white matter and subsequent evolution of secondary pathogenic events at the cellular, biochemical, and molecular levels, which collectively mediate widespread neurodegeneration. An emerging field of research addresses brain injuries related to the military, in particular blast-induced brain injuries. What is clear from the effort to date is that the pathobiology of military TBIs, particularly BINT, has characteristics not seen in other types of brain injury, despite similar secondary injury cascades. The pathobiology of primary BINT is extremely complex. It comprises systemic, local, and cerebral responses interacting and often occurring in parallel. Activation of the autonomous nervous system, sudden pressure-increase in vital organs such as lungs and liver, and activation of neuroendocrine-immune system are among the most important mechanisms significantly contributing to molecular changes and cascading injury mechanisms in the brain. PMID- 19809469 TI - Good science and good ethics: why we should discourage payment for eggs for stem cell research. PMID- 19809470 TI - Disorders of nucleotide excision repair: the genetic and molecular basis of heterogeneity. AB - Mutations in genes on the nucleotide excision repair pathway are associated with diseases, such as xeroderma pigmentosum, Cockayne syndrome and trichothiodystrophy, that involve skin cancer and developmental and neurological symptoms. These mutations cause the defective repair of damaged DNA and increased transcription arrest but, except for skin cancer, the links between repair and disease have not been obvious. Widely different clinical syndromes seem to result from mutations in the same gene, even when the mutations result in complete loss of function. The mapping of mutations in recently solved protein structures has begun to clarify the links between the molecular defects and phenotypes, but the identification of additional sources of clinical variability is still necessary. PMID- 19809471 TI - The functional plasticity of T cell subsets. AB - In 1986, Robert Coffman and Timothy Mossman first described the division of CD4(+) T cells into functional subsets, termed T helper 1 (T(H)1) and T(H)2, based on cytokine production, and in doing so unwittingly opened a Pandora's box of complexity and controversy. Although the mechanisms that regulate T(H)1 and T(H)2 cells are now well known, recent descriptions of other CD4(+) T cell subsets--such as regulatory T cells, T follicular helper cells, T(H)17, T(H)22 and most recently T(H)9 and T(H)22 cells--have questioned how we think of T cell subsets and what commitment to a functional T cell subset means. Here, Nature Reviews Immunology asks four leaders in the field their thoughts on the functional plasticity of T cell subsets. PMID- 19809472 TI - Long-term potentiation: two pathways meet at neurogranin. PMID- 19809473 TI - High-resolution SNP arrays in mental retardation diagnostics: how much do we gain? AB - We used Affymetrix 6.0 GeneChip SNP arrays to characterize copy number variations (CNVs) in a cohort of 70 patients previously characterized on lower-density oligonucleotide arrays affected by idiopathic mental retardation and dysmorphic features. The SNP array platform includes approximately 900,000 SNP probes and 900,000 non-SNP oligonucleotide probes at an average distance of 0.7 Kb, which facilitates coverage of the whole genome, including coding and noncoding regions. The high density of probes is critical for detecting small CNVs, but it can lead to data interpretation problems. To reduce the number of false positives, parameters were set to consider only imbalances >75 Kb encompassing at least 80 probe sets. The higher resolution of the SNP array platform confirmed the increased ability to detect small CNVs, although more than 80% of these CNVs overlapped to copy number 'neutral' polymorphism regions and 4.4% of them did not contain known genes. In our cohort of 70 patients, of the 51 previously evaluated as 'normal' on the Agilent 44K array, the SNP array platform disclosed six additional CNV changes, including three in three patients, which may be pathogenic. This suggests that about 6% of individuals classified as 'normal' using the lower-density oligonucleotide array could be found to be affected by a genomic disorder when evaluated with the higher-density microarray platforms. PMID- 19809474 TI - Study of smell and reproductive organs in a mouse model for CHARGE syndrome. AB - CHARGE syndrome is a multiple congenital anomaly syndrome characterised by Coloboma, Heart defects, Atresia of choanae, Retardation of growth and/or development, Genital hypoplasia, and Ear anomalies often associated with deafness. It is caused by heterozygous mutations in the CHD7 gene and shows a highly variable phenotype. Anosmia and hypogonadotropic hypogonadism occur in the majority of the CHARGE patients, but the underlying pathogenesis is unknown. Therefore, we studied the ability to smell and aspects of the reproductive system (reproductive performance, gonadotropin-releasing hormone (GnRH) neurons and anatomy of testes and uteri) in a mouse model for CHARGE syndrome, the whirligig mouse (Chd7(Whi/+)). We showed that Chromodomain Helicase DNA-binding protein 7 (Chd7) is expressed in brain areas involved in olfaction and reproduction during embryonic development. We observed poorer performance in the smell test in adult Chd7(Whi/+) mice, secondary either to olfactory dysfunction or to balance disturbances. Olfactory bulb and reproductive organ abnormalities were observed in a proportion of Chd7(Whi/+) mice. Hypothalamic GnRH neurons were slightly reduced in Chd7(Whi/+) females and reproductive performance was slightly less in Chd7(Whi/+) mice. This study shows that the penetrance of anosmia and hypogonadotropic hypogonadism is lower in Chd7(Whi/+) mice than in CHARGE patients. Interestingly, many phenotypic features of the Chd7 mutation showed incomplete penetrance in our model mice, despite the use of inbred, genetically identical mice. This supports the theory that the extreme variability of the CHARGE phenotype in both humans and mice might be attributed to variations in the fetal microenvironment or to purely stochastic events. PMID- 19809476 TI - A powerful genome-wide feasible approach to detect parent-of-origin effects in studies of quantitative traits. AB - There is currently a lot of interest in the role of genomic imprinting in mammalian development. Many human diseases, such as cancer, obesity, diabetes and behavioral traits, may be related to imprinted genes. When searching for genes related to complex disorders, the power of genome-wide association analysis can be improved by introducing parent-of-origin effects into the analyses. For quantitative traits, family-based TDT analysis has successfully implemented such an approach. Although attractive for several reasons, TDT-based tests are known to be less powerful than methods based on measured genotype approaches. In this study, we describe a fast, powerful method for detecting parent-of-origin effects in studies of quantitative traits using a measured genotype framework. First, for each locus studied, we estimate the probabilities of an allele's parental origin using multipoint haplotype reconstruction. Next, we introduce the parental origin of these alleles as a covariate in regression models during the second step of GRAMMAR, a fast approximation to the measured genotype approach. We show that, compared with a TDT-based analysis, our method has a higher power to detect a locus exhibiting a parent-of-origin effect. Moreover, our method is applicable to a wider range of data, including pedigree structures that are not very informative for TDT. The method gives no false positives in the absence of parent of-origin effects, under both additive and dominant models. As this method is an extension of the rapid GRAMMAR analysis, it is fast enough to be suitable for genome-wide association scans. PMID- 19809477 TI - Progress in therapeutic antisense applications for neuromuscular disorders. AB - Neuromuscular disorders are a frequent cause of chronic disability in man. They often result from mutations in single genes and are thus, in principle, well suited for gene therapy. However, the tissues involved (muscle and the central nervous system) are post-mitotic, which poses a challenge for most viral vectors. In some cases, alternative approaches may use small molecules, for example, antisense oligonucleotides (AONs). These do not deliver a new gene, but rather modulate existing gene products or alter the utilization of pathways. For Duchenne muscular dystrophy, this approach is in early phase clinical trials, and for two other common neuromuscular disorders (spinal muscular atrophy and myotonic dystrophy), significant preclinical advances have recently been made. PMID- 19809478 TI - Functional consequences of mitochondrial tRNA Trp and tRNA Arg mutations causing combined OXPHOS defects. AB - Combined oxidative phosphorylation (OXPHOS) system deficiencies are a group of mitochondrial disorders that are associated with a range of clinical phenotypes and genetic defects. They occur in approximately 30% of all OXPHOS disorders and around 4% are combined complex I, III and IV deficiencies. In this study we present two mutations in the mitochondrial tRNA(Trp) (MT-TW) and tRNA(Arg) (MT TR) genes, m.5556G>A and m.10450A>G, respectively, which were detected in two unrelated patients showing combined OXPHOS complex I, III and IV deficiencies and progressive multisystemic diseases. Both mitochondrial tRNA mutations were almost homoplasmic in fibroblasts and muscle tissue of the two patients and not present in controls. Patient fibroblasts showed a general mitochondrial translation defect. The mutations resulted in lowered steady-state levels and altered conformations of the tRNAs. Cybrid cell lines showed similar tRNA defects and impairment of OXPHOS complex assembly as patient fibroblasts. Our results show that these tRNA(Trp) and tRNA(Arg) mutations cause the combined OXPHOS deficiencies in the patients, adding to the still expanding group of pathogenic mitochondrial tRNA mutations. PMID- 19809479 TI - Design and evaluation of a panel of single-nucleotide polymorphisms in microRNA genomic regions for association studies in human disease. AB - MicroRNAs (miRNA) are recognized posttranscriptional gene repressors involved in the control of almost every biological process. Allelic variants in these regions may be an important source of phenotypic diversity and contribute to disease susceptibility. We analyzed the genomic organization of 325 human miRNAs (release 7.1, miRBase) to construct a panel of 768 single-nucleotide polymorphisms (SNPs) covering approximately 1 Mb of genomic DNA, including 131 isolated miRNAs (40%) and 194 miRNAs arranged in 48 miRNA clusters, as well as their 5-kb flanking regions. Of these miRNAs, 37% were inside known protein-coding genes, which were significantly associated with biological functions regarding neurological, psychological or nutritional disorders. SNP coverage analysis revealed a lower SNP density in miRNAs compared with the average of the genome, with only 24 SNPs located in the 325 miRNAs studied. Further genotyping of 340 unrelated Spanish individuals showed that more than half of the SNPs in miRNAs were either rare or monomorphic, in agreement with the reported selective constraint on human miRNAs. A comparison of the minor allele frequencies between Spanish and HapMap population samples confirmed the applicability of this SNP panel to the study of complex disorders among the Spanish population, and revealed two miRNA regions, hsa-mir-26a-2 in the CTDSP2 gene and hsa-mir-128-1 in the R3HDM1 gene, showing geographical allelic frequency variation among the four HapMap populations, probably because of differences in natural selection. The designed miRNA SNP panel could help to identify still hidden links between miRNAs and human disease. PMID- 19809480 TI - Traces of sub-Saharan and Middle Eastern lineages in Indian Muslim populations. AB - Islam is the second most practiced religion in India, next to Hinduism. It is still unclear whether the spread of Islam in India has been only a cultural transformation or is associated with detectable levels of gene flow. To estimate the contribution of West Asian and Arabian admixture to Indian Muslims, we assessed genetic variation in mtDNA, Y-chromosomal and LCT/MCM6 markers in 472, 431 and 476 samples, respectively, representing six Muslim communities from different geographical regions of India. We found that most of the Indian Muslim populations received their major genetic input from geographically close non Muslim populations. However, low levels of likely sub-Saharan African, Arabian and West Asian admixture were also observed among Indian Muslims in the form of L0a2a2 mtDNA and E1b1b1a and J(*)(xJ2) Y-chromosomal lineages. The distinction between Iranian and Arabian sources was difficult to make with mtDNA and the Y chromosome, as the estimates were highly correlated because of similar gene pool compositions in the sources. In contrast, the LCT/MCM6 locus, which shows a clear distinction between the two sources, enabled us to rule out significant gene flow from Arabia. Overall, our results support a model according to which the spread of Islam in India was predominantly cultural conversion associated with minor but still detectable levels of gene flow from outside, primarily from Iran and Central Asia, rather than directly from the Arabian Peninsula. PMID- 19809481 TI - Prader-Willi and Angelman syndromes: genetic counseling. PMID- 19809482 TI - Understanding sickle cell carrier status identified through newborn screening: a qualitative study. AB - The expansion of newborn screening (NBS) is increasing the generation of incidental results, notably carrier results. Although carrier status is generally understood to be clinically benign, concerns persist that parents may misunderstand its meaning, with deleterious effects on children and their families. Expansion of the NBS panel in Ontario, Canada in 2006 to include sickle cell disorders drew attention to the policy challenge of incidental carrier results. We conducted a study of consumer and provider attitudes to inform policy on disclosure. In this paper, we report the results of (i) qualitative interviews with health-care providers, advocates and parents of carrier infants and (ii) focus groups with new parents and individuals active with the sickle cell community. Lay and provider participants generally believed that carrier results were clinically insignificant. However, some uncertainty persisted among lay consumers in the form of conjecture or doubt. In addition, consumers and advocates who were most informed about the disease articulated insistent yet dissonant claims of clinical significance. Meanwhile, providers referenced research knowledge to offer an equivocal assessment of the possibility and significance of clinically symptomatic carrier status. We conclude that many interpretations of carrier status are in circulation, failing to fit neatly into the categories of 'clinically significant' or 'benign.' This creates challenges for communicating clearly with parents - challenges exacerbated by inconsistent messages from screening programs regarding the significance of sickle cell carrier status. Disclosure policy related to incidentally generated infant carrier results needs to account for these complex realities. PMID- 19809483 TI - Comparison of human chromosome 19q13 and syntenic region on mouse chromosome 7 reveals absence, in man, of 11.6 Mb containing four mouse calcium-sensing receptor-related sequences: relevance to familial benign hypocalciuric hypercalcaemia type 3. AB - Familial benign hypocalciuric hypercalcaemia (FBHH) is a genetically heterogeneous disorder that consists of three designated types, FBHH1, FBHH2 and FBHH3, whose chromosomal locations are 3q21.1, 19p and 19q13, respectively. FBHH1 is caused by mutations of a calcium-sensing receptor (CaSR), but the abnormalities underlying FBHH2 and FBHH3 are unknown. FBHH3, also referred to as the Oklahoma variant (FBHH(Ok)), has been mapped to a 12cM interval, flanked by D19S908 and D19S866. To refine the location of FBHH3, we pursued linkage studies using 24 polymorphic loci. Our results establish a linkage between FBHH3 and 17 of these loci, and indicate that FBHH3 is located in a 4.1 Mb region flanked centromerically by D19S112 and telomerically by rs245111, which in the syntenic region on mouse chromosome 7 contains four Casr-related sequences (Gprc2a-rss). However, human homologues of these Gprc2a-rss were not found and a comparative analysis of the 22.0 Mb human and 39.3 Mb mouse syntenic regions showed evolutionary conservation of two segments that were inverted with loss from the human genome of 11.6 Mb that contained the four Gprc2a-rss. Thus, FBHH3 cannot be attributed to Gprc2a-rss abnormalities. DNA sequence analysis of 12 other genes from the interval that were expressed in the parathyroids and/or kidneys did not detect any abnormalities, thereby indicating that these genes are unlikely to be the cause of FBHH3. The results of this study have refined the map location of FBHH3, which will facilitate the identification of another CaSR or a mediator of calcium homeostasis. PMID- 19809484 TI - The 2q23.1 microdeletion syndrome: clinical and behavioural phenotype. AB - Six submicroscopic deletions comprising chromosome band 2q23.1 in patients with severe mental retardation (MR), short stature, microcephaly and epilepsy have been reported, suggesting that haploinsufficiency of one or more genes in the 2q23.1 region might be responsible for the common phenotypic features in these patients. In this study, we report the molecular and clinical characterisation of nine new 2q23.1 deletion patients and a clinical update on two previously reported patients. All patients were mentally retarded with pronounced speech delay and additional abnormalities including short stature, seizures, microcephaly and coarse facies. The majority of cases presented with stereotypic repetitive behaviour, a disturbed sleep pattern and a broad-based gait. These features led to the initial clinical impression of Angelman, Rett or Smith Magenis syndromes in several patients. The overlapping 2q23.1 deletion region in all 15 patients comprises only one gene, namely, MBD5. Interestingly, MBD5 is a member of the methyl CpG-binding domain protein family, which also comprises MECP2, mutated in Rett's syndrome. Another gene in the 2q23.1 region, EPC2, was deleted in 12 patients who had a broader phenotype than those with a deletion of MBD5 only. EPC2 is a member of the polycomb protein family, involved in heterochromatin formation and might be involved in causing MR. Patients with a 2q23.1 microdeletion present with a variable phenotype and the diagnosis should be considered in mentally retarded children with coarse facies, seizures, disturbed sleeping patterns and additional specific behavioural problems. PMID- 19809485 TI - Attitudes toward genetic testing in childhood and reproductive decision-making for familial adenomatous polyposis. AB - Childhood DNA testing, prenatal diagnosis (PND) and preimplantation genetic diagnosis (PGD) are available for familial adenomatous polyposis (FAP). However, the use of PND and PGD is controversial. The purpose of this study was to investigate attitudes toward, and experiences with, childhood DNA testing, PND and PGD among members of families at high risk for FAP. In this nationwide, cross sectional study, questionnaires were sent to individuals from families at high risk for FAP assessing attitudes toward and experiences with childhood testing, PND and PGD, as well as several sociodemographic, clinical and psychosocial variables. Of the individuals from FAP families invited to participate in the study, 525 members participated (response rate=64%). Most parents who had children who were minors (n=93) (82%) were satisfied with the DNA testing procedure. One-third of all individuals wanted DNA testing for their children before age 12. Forty percent of FAP patients indicated that the disease influenced their desire to have children. Only 15% considered termination of pregnancy for FAP acceptable. Approximately 30% of individuals with a FAP diagnosis and their partners considered PND and PGD as acceptable for themselves. A positive attitude was associated with higher levels of guilt and a positive attitude toward termination of pregnancy. Importantly, of those with FAP at childbearing age, 84% had had no previous information at all about either PND or PGD. Future efforts should be aimed at educating FAP family members about reproductive options, allowing them to make an informed choice about family planning. Routine discussion of all reproductive options with a medical specialist should be encouraged. PMID- 19809486 TI - Comprehensive expression analysis of FSHD candidate genes at the mRNA and protein level. AB - In facioscapulohumeral muscular dystrophy (FSHD) the majority of patients carry a D4Z4 macrosatellite repeat contraction in the subtelomere of chromosome 4q. Several disease mechanisms have been proposed to explain how repeat contraction causes muscular dystrophy. All proposed mechanisms foresee a change from a closed to a more open chromatin structure followed by loss of control over expression of genes in or proximal to D4Z4. Initially, a distance and residual repeat size dependent upregulation of the candidate genes FRG2, FRG1 and ANT1 was observed, but most successive expression studies failed to support transcriptional upregulation of 4qter genes. Moreover, chromatin studies do not provide evidence for a cis-spreading mechanism operating at 4qter in FSHD. In part, this inconsistency may be explained by differences in the techniques used, and the use of RNA samples obtained from different muscle groups. The aim of this study is to comprehensively and uniformly study the expression of the FSHD candidate genes FRG1, FRG2, CRYM, ANT1, ALP, PITX1 and LRP2BP at the RNA and protein level in identically processed primary myoblasts, myotubes and quadriceps muscle. Expression was compared between samples obtained from FSHD patients and normal controls with samples from myotonic dystrophy type 1 patients as disease controls. No consistent changes in RNA or protein expression levels were observed between the samples. The one exception was a selective increase in FRG2 mRNA expression in FSHD myotubes. This study provides further evidence that there is no demonstrable consistent, large magnitude, overexpression of any of the FSHD candidate genes. PMID- 19809487 TI - London's hottest new attraction is... scientists at work. PMID- 19809488 TI - PORE-ing over ERK substrates. PMID- 19809489 TI - Trigger factor finds new jobs and contacts. PMID- 19809490 TI - Silence of the budding yeast. PMID- 19809492 TI - Telomeric circles: universal players in telomere maintenance? AB - To maintain linear DNA genomes, organisms have evolved numerous means of solving problems associated with DNA ends (telomeres), including telomere-associated retrotransposons, palindromes, hairpins, covalently bound proteins and the addition of arrays of simple DNA repeats. Telomeric arrays can be maintained through various mechanisms such as telomerase activity or recombination. The recombination-dependent maintenance pathways may include telomeric loops (t loops) and telomeric circles (t-circles). The potential involvement of t-circles in telomere maintenance was first proposed for linear mitochondrial genomes. The occurrence of t-circles in a wide range of organisms, spanning yeasts, plants and animals, suggests the involvement of t-circles in many phenomena including the alternative-lengthening of telomeres (ALT) pathway and telomere rapid deletion (TRD). In this Perspective, we summarize these findings and discuss how t-circles may be related to t-loops and how t-circles may have initiated the evolution of telomeres. PMID- 19809493 TI - Stem cell therapy with overexpressed VEGF and PDGF genes improves cardiac function in a rat infarct model. AB - BACKGROUND: Therapeutic potential was evaluated in a rat model of myocardial infarction using nanofiber-expanded human cord blood derived hematopoietic stem cells (CD133+/CD34+) genetically modified with VEGF plus PDGF genes (VIP). METHODS AND FINDINGS: Myocardial function was monitored every two weeks up to six weeks after therapy. Echocardiography revealed time dependent improvement of left ventricular function evaluated by M-mode, fractional shortening, anterior wall tissue velocity, wall motion score index, strain and strain rate in animals treated with VEGF plus PDGF overexpressed stem cells (VIP) compared to nanofiber expanded cells (Exp), freshly isolated cells (FCB) or media control (Media). Improvement observed was as follows: VIP>Exp> FCB>media. Similar trend was noticed in the exercise capacity of rats on a treadmill. These findings correlated with significantly increased neovascularization in ischemic tissue and markedly reduced infarct area in animals in the VIP group. Stem cells in addition to their usual homing sites such as lung, spleen, bone marrow and liver, also migrated to sites of myocardial ischemia. The improvement of cardiac function correlated with expression of heart tissue connexin 43, a gap junctional protein, and heart tissue angiogenesis related protein molecules like VEGF, pNOS3, NOS2 and GSK3. There was no evidence of upregulation in the molecules of oncogenic potential in genetically modified or other stem cell therapy groups. CONCLUSION: Regenerative therapy using nanofiber-expanded hematopoietic stem cells with overexpression of VEGF and PDGF has a favorable impact on the improvement of rat myocardial function accompanied by upregulation of tissue connexin 43 and pro angiogenic molecules after infarction. PMID- 19809494 TI - The roles and acting mechanism of Caenorhabditis elegans DNase II genes in apoptotic dna degradation and development. AB - DNase II enzymes are acidic endonucleases that have been implicated in mediating apoptotic DNA degradation, a critical cell death execution event. C. elegans genome contains three DNase II homologues, NUC-1, CRN-6, and CRN-7, but their expression patterns, acting sites, and roles in apoptotic DNA degradation and development are unclear. We have conducted a comprehensive analysis of three C. elegans DNase II genes and found that nuc-1 plays a major role, crn-6 plays an auxiliary role, and crn-7 plays a negligible role in resolving 3' OH DNA breaks generated in apoptotic cells. Promoter swapping experiments suggest that crn-6 but not crn-7 can partially substitute for nuc-1 in mediating apoptotic DNA degradation and both fail to replace nuc-1 in degrading bacterial DNA in intestine. Despite of their restricted and largely non-overlapping expression patterns, both CRN-6 and NUC-1 can mediate apoptotic DNA degradation in many cells, suggesting that they are likely secreted nucleases that are retaken up by other cells to exert DNA degradation functions. Removal or disruption of NUC-1 secretion signal eliminates NUC-1's ability to mediate DNA degradation across its expression border. Furthermore, blocking cell corpse engulfment does not affect apoptotic DNA degradation mediated by nuc-1, suggesting that NUC-1 acts in apoptotic cells rather than in phagocytes to resolve 3' OH DNA breaks. Our study illustrates how multiple DNase II nucleases play differential roles in apoptotic DNA degradation and development and reveals an unexpected mode of DNase II action in mediating DNA degradation. PMID- 19809495 TI - Aldo keto reductase 1B7 and prostaglandin F2alpha are regulators of adrenal endocrine functions. AB - Prostaglandin F(2alpha) (PGF(2alpha)), represses ovarian steroidogenesis and initiates parturition in mammals but its impact on adrenal gland is unknown. Prostaglandins biosynthesis depends on the sequential action of upstream cyclooxygenases (COX) and terminal synthases but no PGF(2alpha) synthases (PGFS) were functionally identified in mammalian cells. In vitro, the most efficient mammalian PGFS belong to aldo-keto reductase 1B (AKR1B) family. The adrenal gland is a major site of AKR1B expression in both human (AKR1B1) and mouse (AKR1B3, AKR1B7). Thus, we examined the PGF(2alpha) biosynthetic pathway and its functional impact on both cortical and medullary zones. Both compartments produced PGF(2alpha) but expressed different biosynthetic isozymes. In chromaffin cells, PGF(2alpha) secretion appeared constitutive and correlated to continuous expression of COX1 and AKR1B3. In steroidogenic cells, PGF(2alpha) secretion was stimulated by adrenocorticotropic hormone (ACTH) and correlated to ACTH responsiveness of both COX2 and AKR1B7/B1. The pivotal role of AKR1B7 in ACTH induced PGF(2alpha) release and functional coupling with COX2 was demonstrated using over- and down-expression in cell lines. PGF(2alpha) receptor was only detected in chromaffin cells, making medulla the primary target of PGF(2alpha) action. By comparing PGF(2alpha)-responsiveness of isolated cells and whole adrenal cultures, we demonstrated that PGF(2alpha) repressed glucocorticoid secretion by an indirect mechanism involving a decrease in catecholamine release which in turn decreased adrenal steroidogenesis. PGF(2alpha) may be regarded as a negative autocrine/paracrine regulator within a novel intra-adrenal feedback loop. The coordinated cell-specific regulation of COX2 and AKR1B7 ensures the generation of this stress-induced corticostatic signal. PMID- 19809496 TI - Functional genetic variants in DC-SIGNR are associated with mother-to-child transmission of HIV-1. AB - BACKGROUND: Mother-to-child transmission (MTCT) is the main cause of HIV-1 infection in children worldwide. Given that the C-type lectin receptor, dendritic cell-specific ICAM-grabbing non-integrin-related (DC-SIGNR, also known as CD209L or liver/lymph node-specific ICAM-grabbing non-integrin (L-SIGN)), can interact with pathogens including HIV-1 and is expressed at the maternal-fetal interface, we hypothesized that it could influence MTCT of HIV-1. METHODS AND FINDINGS: To investigate the potential role of DC-SIGNR in MTCT of HIV-1, we carried out a genetic association study of DC-SIGNR in a well-characterized cohort of 197 HIV infected mothers and their infants recruited in Harare, Zimbabwe. Infants harbouring two copies of DC-SIGNR H1 and/or H3 haplotypes (H1-H1, H1-H3, H3-H3) had a 3.6-fold increased risk of in utero (IU) (P = 0.013) HIV-1 infection and a 5.7-fold increased risk of intrapartum (IP) (P = 0.025) HIV-1 infection after adjusting for a number of maternal factors. The implicated H1 and H3 haplotypes share two single nucleotide polymorphisms (SNPs) in promoter region (p-198A) and intron 2 (int2-180A) that were associated with increased risk of both IU (P = 0.045 and P = 0.003, respectively) and IP (P = 0.025, for int2-180A) HIV-1 infection. The promoter variant reduced transcriptional activity in vitro. In homozygous H1 infants bearing both the p-198A and int2-180A mutations, we observed a 4-fold decrease in the level of placental DC-SIGNR transcripts, disproportionately affecting the expression of membrane-bound isoforms compared to infant noncarriers (P = 0.011). CONCLUSION: These results suggest that DC SIGNR plays a crucial role in MTCT of HIV-1 and that impaired placental DC-SIGNR expression increases risk of transmission. PMID- 19809497 TI - From the eye of the albatrosses: a bird-borne camera shows an association between albatrosses and a killer whale in the Southern Ocean. AB - Albatrosses fly many hundreds of kilometers across the open ocean to find and feed upon their prey. Despite the growing number of studies concerning their foraging behaviour, relatively little is known about how albatrosses actually locate their prey. Here, we present our results from the first deployments of a combined animal-borne camera and depth data logger on free-ranging black-browed albatrosses (Thalassarche melanophrys). The still images recorded from these cameras showed that some albatrosses actively followed a killer whale (Orcinus orca), possibly to feed on food scraps left by this diving predator. The camera images together with the depth profiles showed that the birds dived only occasionally, but that they actively dived when other birds or the killer whale were present. This association with diving predators or other birds may partially explain how albatrosses find their prey more efficiently in the apparently 'featureless' ocean, with a minimal requirement for energetically costly diving or landing activities. PMID- 19809498 TI - An ATP and oxalate generating variant tricarboxylic acid cycle counters aluminum toxicity in Pseudomonas fluorescens. AB - Although the tricarboxylic acid (TCA) cycle is essential in almost all aerobic organisms, its precise modulation and integration in global cellular metabolism is not fully understood. Here, we report on an alternative TCA cycle uniquely aimed at generating ATP and oxalate, two metabolites critical for the survival of Pseudomonas fluorescens. The upregulation of isocitrate lyase (ICL) and acylating glyoxylate dehydrogenase (AGODH) led to the enhanced synthesis of oxalate, a dicarboxylic acid involved in the immobilization of aluminum (Al). The increased activity of succinyl-CoA synthetase (SCS) and oxalate CoA-transferase (OCT) in the Al-stressed cells afforded an effective route to ATP synthesis from oxalyl CoA via substrate level phosphorylation. This modified TCA cycle with diminished efficacy in NADH production and decreased CO(2)-evolving capacity, orchestrates the synthesis of oxalate, NADPH, and ATP, ingredients pivotal to the survival of P. fluorescens in an Al environment. The channeling of succinyl-CoA towards ATP formation may be an important function of the TCA cycle during anaerobiosis, Fe starvation and O(2)-limited conditions. PMID- 19809499 TI - Catechol-o-methyltransferase expression and 2-methoxyestradiol affect microtubule dynamics and modify steroid receptor signaling in leiomyoma cells. AB - CONTEXT: Development of optimal medicinal treatments of uterine leiomyomas represents a significant challenge. 2-Methoxyestradiol (2ME) is an endogenous estrogen metabolite formed by sequential action of CYP450s and catechol-O methyltransferase (COMT). Our previous study demonstrated that 2ME is a potent antiproliferative, proapoptotic, antiangiogenic, and collagen synthesis inhibitor in human leiomyomas cells (huLM). OBJECTIVES: Our objectives were to investigate whether COMT expression, by the virtue of 2ME formation, affects the growth of huLM, and to explore the cellular and molecular mechanisms whereby COMT expression or treatment with 2ME affect these cells. RESULTS: Our data demonstrated that E(2)-induced proliferation was less pronounced in cells over expressing COMT or treated with 2ME (500 nM). This effect on cell proliferation was associated with microtubules stabilization and diminution of estrogen receptor alpha (ERalpha) and progesterone receptor (PR) transcriptional activities, due to shifts in their subcellular localization and sequestration in the cytoplasm. In addition, COMT over expression or treatment with 2ME reduced the expression of hypoxia-inducible factor -1alpha (HIF-1 alpha) and the basal level as well as TNF-alpha-induced aromatase (CYP19) expression. CONCLUSIONS: COMT over expression or treatment with 2ME stabilize microtubules, ameliorates E(2)-induced proliferation, inhibits ERalpha and PR signaling, and reduces HIF-1 alpha and CYP19 expression in human uterine leiomyoma cells. Thus, microtubules are a candidate target for treatment of uterine leiomyomas. In addition, the naturally occurring microtubule-targeting agent 2ME represents a potential new therapeutic for uterine leiomyomas. PMID- 19809500 TI - Edge detection in landing budgerigars (Melopsittacus undulatus). AB - BACKGROUND: While considerable scientific effort has been devoted to studying how birds navigate over long distances, relatively little is known about how targets are detected, obstacles are avoided and smooth landings are orchestrated. Here we examine how visual features in the environment, such as contrasting edges, determine where a bird will land. METHODOLOGY/PRINCIPAL FINDINGS: Landing in budgerigars (Melopsittacus undulatus) was investigated by training them to fly from a perch to a feeder, and video-filming their landings. The feeder was placed on a grey disc that produced a contrasting edge against a uniformly blue background. We found that the birds tended to land primarily at the edge of the disc and walk to the feeder, even though the feeder was in the middle of the disc. This suggests that the birds were using the visual contrast at the boundary of the disc to target their landings. When the grey level of the disc was varied systematically, whilst keeping the blue background constant, there was one intermediate grey level at which the budgerigar's preference for the disc boundary disappeared. The budgerigars then landed randomly all over the test surface. Even though this disc is (for humans) clearly distinguishable from the blue background, it offers very little contrast against the background, in the red and green regions of the spectrum. CONCLUSIONS: We conclude that budgerigars use visual edges to target and guide landings. Calculations of photoreceptor excitation reveal that edge detection in landing budgerigars is performed by a color-blind luminance channel that sums the signals from the red and green photoreceptors, or, alternatively, receives input from the red double-cones. This finding has close parallels to vision in honeybees and primates, where edge detection and motion perception are also largely color-blind. PMID- 19809501 TI - Antimicrobial activity of human prion protein is mediated by its N-terminal region. AB - BACKGROUND: Cellular prion-related protein (PrP(c)) is a cell-surface protein that is ubiquitously expressed in the human body. The multifunctionality of PrP(c), and presence of an exposed cationic and heparin-binding N-terminus, a feature characterizing many antimicrobial peptides, made us hypothesize that PrP(c) could exert antimicrobial activity. METHODOLOGY AND PRINCIPAL FINDINGS: Intact recombinant PrP exerted antibacterial and antifungal effects at normal and low pH. Studies employing recombinant PrP and N- and C-terminally truncated variants, as well as overlapping peptide 20mers, demonstrated that the antimicrobial activity is mediated by the unstructured N-terminal part of the protein. Synthetic peptides of the N-terminus of PrP killed the Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa, and the Gram-positive Bacillus subtilis and Staphylococcus aureus, as well as the fungus Candida parapsilosis. Fluorescence studies of peptide-treated bacteria, paired with analysis of peptide effects on liposomes, showed that the peptides exerted membrane-breaking effects similar to those seen after treatment with the "classical" human antimicrobial peptide LL-37. In contrast to LL-37, however, no marked helix induction was detected for the PrP-derived peptides in presence of negatively charged (bacteria-mimicking) liposomes. PrP furthermore showed an inducible expression during wounding of human skin ex vivo and in vivo, as well as stimulation of keratinocytes with TGF-alpha in vitro. CONCLUSIONS: The demonstration of an antimicrobial activity of PrP, localisation of its activity to the N-terminal and heparin-binding region, combined with results showing an increased expression of PrP during wounding, indicate that PrPs could have a previously undisclosed role in host defense. PMID- 19809502 TI - Long-Lasting Insecticidal Hammocks for controlling forest malaria: a community based trial in a rural area of central Vietnam. AB - BACKGROUND: In Vietnam, malaria remains a problem in some remote areas located along its international borders and in the central highlands, partly due to the bionomics of the local vector, mainly found in forested areas and less vulnerable to standard control measures. Long Lasting Insecticidal Hammocks (LLIH), a tailored and user-friendly tool for forest workers, may further contribute in reducing the malaria burden. Their effectiveness was tested in a large community based intervention trial carried out in Ninh Thuan province in Central Vietnam. METHODS AND FINDINGS: Thirty villages (population 18,646) were assembled in 20 clusters (1,000 individuals per cluster) that were randomly allocated to either the intervention or control group (no LLIH) after stratification according to the pre-intervention P. falciparum antibody prevalence (<30%; > or =30%). LLIH were distributed to the intervention group in December 2004. For the following 2 years, the incidence of clinical malaria and the prevalence of infection were determined by passive case detection at community level and by bi-annual malariometric surveys. A 2-fold larger effect on malaria incidence in the intervention as compared to the control group was observed. Similarly, malaria prevalence decreased more substantially in the intervention (1.6-fold greater reduction) than in the control group. Both for incidence and prevalence, a stronger and earlier effect of the intervention was observed in the high endemicity stratum. The number of malaria cases and infections averted by the intervention overall was estimated at 10.5 per 1,000 persons and 5.6/100 individuals, respectively, for the last half of 2006. In the high endemicity stratum, the impact was much higher, i.e. 29/1000 malaria cases and 15.7 infections/100 individuals averted. CONCLUSIONS: LLIH reduced malaria incidence and prevalence in this remote and forested area of Central Vietnam. As the targets of the newly-launched Global Malaria Action Plan include the 75% reduction of the global malaria cases by 2015 and eventually the elimination/eradication of malaria in the long term, LLIH may represent an additional tool for reaching such objectives, particularly in high endemicity areas where standard control tools have a modest impact, such as in remote and forested areas of Southeast Asia and possibly South America. TRIAL REGISTRATION: ClinicalTrials.gov NCT00853281. PMID- 19809503 TI - New insights in the contribution of voltage-gated Na(v) channels to rat aorta contraction. AB - BACKGROUND: Despite increasing evidence for the presence of voltage-gated Na(+) channels (Na(v)) isoforms and measurements of Na(v) channel currents with the patch-clamp technique in arterial myocytes, no information is available to date as to whether or not Na(v) channels play a functional role in arteries. The aim of the present work was to look for a physiological role of Na(v) channels in the control of rat aortic contraction. METHODOLOGY/PRINCIPAL FINDINGS: Na(v) channels were detected in the aortic media by Western blot analysis and double immunofluorescence labeling for Na(v) channels and smooth muscle alpha-actin using specific antibodies. In parallel, using real time RT-PCR, we identified three Na(v) transcripts: Na(v)1.2, Na(v)1.3, and Na(v)1.5. Only the Na(v)1.2 isoform was found in the intact media and in freshly isolated myocytes excluding contamination by other cell types. Using the specific Na(v) channel agonist veratridine and antagonist tetrodotoxin (TTX), we unmasked a contribution of these channels in the response to the depolarizing agent KCl on rat aortic isometric tension recorded from endothelium-denuded aortic rings. Experimental conditions excluded a contribution of Na(v) channels from the perivascular sympathetic nerve terminals. Addition of low concentrations of KCl (2-10 mM), which induced moderate membrane depolarization (e.g., from -55.9+/-1.4 mV to 45.9+/-1.2 mV at 10 mmol/L as measured with microelectrodes), triggered a contraction potentiated by veratridine (100 microM) and blocked by TTX (1 microM). KB-R7943, an inhibitor of the reverse mode of the Na(+)/Ca(2+) exchanger, mimicked the effect of TTX and had no additive effect in presence of TTX. CONCLUSIONS/SIGNIFICANCE: These results define a new role for Na(v) channels in arterial physiology, and suggest that the TTX-sensitive Na(v)1.2 isoform, together with the Na(+)/Ca(2+) exchanger, contributes to the contractile response of aortic myocytes at physiological range of membrane depolarization. PMID- 19809504 TI - Reassortment patterns in Swine influenza viruses. AB - Three human influenza pandemics occurred in the twentieth century, in 1918, 1957, and 1968. Influenza pandemic strains are the results of emerging viruses from non human reservoirs to which humans have little or no immunity. At least two of these pandemic strains, in 1957 and in 1968, were the results of reassortments between human and avian viruses. Also, many cases of swine influenza viruses have reportedly infected humans, in particular, the recent H1N1 influenza virus of swine origin, isolated in Mexico and the United States. Pigs are documented to allow productive replication of human, avian, and swine influenza viruses. Thus it has been conjectured that pigs are the "mixing vessel" that create the avian human reassortant strains, causing the human pandemics. Hence, studying the process and patterns of viral reassortment, especially in pigs, is a key to better understanding of human influenza pandemics. In the last few years, databases containing sequences of influenza A viruses, including swine viruses, collected since 1918 from diverse geographical locations, have been developed and made publicly available. In this paper, we study an ensemble of swine influenza viruses to analyze the reassortment phenomena through several statistical techniques. The reassortment patterns in swine viruses prove to be similar to the previous results found in human viruses, both in vitro and in vivo, that the surface glycoprotein coding segments reassort most often. Moreover, we find that one of the polymerase segments (PB1), reassorted in the strains responsible for the last two human pandemics, also reassorts frequently. PMID- 19809505 TI - A comparison of biomarker based incidence estimators. AB - BACKGROUND: Cross-sectional surveys utilizing biomarkers that test for recent infection provide a convenient and cost effective way to estimate HIV incidence. In particular, the BED assay has been developed for this purpose. Controversy surrounding the way in which false positive results from the biomarker should be handled has lead to a number of different estimators that account for imperfect specificity. We compare the estimators proposed by McDougal et al., Hargrove et al. and McWalter & Welte. METHODOLOGY/PRINCIPAL FINDINGS: The three estimators are analyzed and compared. An identity showing a relationship between the calibration parameters in the McDougal methodology is shown. When the three estimators are tested under a steady state epidemic, which includes individuals who fail to progress on the biomarker, only the McWalter/Welte method recovers an unbiased result. CONCLUSIONS/SIGNIFICANCE: Our analysis shows that the McDougal estimator can be reduced to a formula that only requires calibration of a mean window period and a long-term specificity. This allows simpler calibration techniques to be used and shows that all three estimators can be expressed using the same set of parameters. The McWalter/Welte method is applicable under the least restrictive assumptions and is the least prone to bias of the methods reviewed. PMID- 19809506 TI - PAR6, a potential marker for the germ cells selected to form primordial follicles in mouse ovary. AB - Partitioning-defective proteins (PAR) are detected to express mainly in the cytoplast, and play an important role in cell polarity. However, we showed here that PAR6, one kind of PAR protein, was localized in the nuclei of mouse oocytes that formed primordial follicles during the perinatal period, suggesting a new role of PAR protein. It is the first time we found that, in mouse fetal ovaries, PAR6 appeared in somatic cell cytoplasm and fell weak when somatic cells invaded germ cell cysts at 17.5 days post coitus (dpc). Meanwhile, the expression of PAR6 was observed in cysts, and became strong in the nuclei of some germ cells at 19.5 dpc and all primordial follicular oocytes at 3 day post parturition (dpp), and then obviously declined when the primordial follicles entered the folliculogenic growth phase. During the primordial follicle pool foundation, the number of PAR6 positive germ cells remained steady and was consistent with that of formed follicles at 3 dpp. There were no TUNEL (apoptosis examination) positive germ cells stained with PAR6 at any time studied. The number of follicles significantly declined when 15.5 dpc ovaries were treated with the anti-PAR6 antibody and PAR6 RNA interference. Carbenoxolone (CBX, a known blocker of gap junctions) inhibited the expression of PAR6 in germ cells and the formation of follicles. Our results suggest that PAR6 could be used as a potential marker of germ cells for the primordial follicle formation, and the expression of PAR6 by a gap junction-dependent process may contribute to the formation of primordial follicles and the maintenance of oocytes at the diplotene stage. PMID- 19809507 TI - CD14 C-159T and toll-like receptor 4 Asp299Gly polymorphisms in surviving meningococcal disease patients. AB - BACKGROUND: Carriage of Neisseria meningitidis occurs approximately in 10% of the population, onset of invasive meningococcal disease (IMD) cannot be predicted and differs between ages. It remains unclear, which host factors determine invasion of the bloodstream by the bacteria. Innate immunity has a very important role in the first recognition of invading pathogens. The functional single nucleotide polymorphisms (SNPs) CD14 C-159T and toll-like receptor 4 (TLR4) Asp299Gly have been associated with the risk of gram-negative infections. However, their role in development of IMD still remains unclear. Our aim was to investigate the influence of CD14 C-159T and TLR4 Asp299Gly polymorphisms on the risk of IMD. METHODOLOGY/PRINCIPAL FINDINGS: It was a retrospective case control study. Surviving Austrian meningococcal disease patients were enrolled by sending buccal swabs for DNA analysis. 185 cases with a proven meningococcal infection and 770 healthy controls were enrolled. In surviving meningococcal disease patients DNA analysis of CD14 C-159T and TLR 4 Asp299Gly polymorphisms was performed, as they are part of the innate immune response to bacterial determinants. CD14 C-159T and TLR4 Asp299Gly SNPs were not significantly associated with the presence of IMD when compared to healthy controls. The odds ratio for CD14 C-159T SNP was 1.14 (95% confidence interval (CI) 0.91-1.43; p = 0.266). In TLR4 Asp 299 Gly SNP the odds ratio was 0.78 (CI 0.47-1.43; p = 0.359). CONCLUSION/SIGNIFICANCE: We could not observe a significant influence of CD14 C-159T and TLR4 Asp299Gly polymorphisms on the risk of developing IMD in surviving meningococcal disease patients. To our knowledge, this is the first study investigating the influence of the CD14 C-159T SNP on the susceptibility to IMD. PMID- 19809508 TI - Dimethylarginine dimethylaminohydrolase-1 transgenic mice are not protected from ischemic stroke. AB - BACKGROUND: Methylated arginines are endogenous analogues of L-arginine, the substrate for nitric oxide (NO) synthase. Asymmetric dimethylarginine (ADMA) interferes with NO formation, causing endothelial dysfunction. ADMA is a predictor of cardiovascular events and mortality in humans. It is eliminated primarily by enzymatic activity of dimethylarginine dimethylaminohydrolase (DDAH). METHODOLOGY/PRINCIPAL FINDINGS: We investigated whether human DDAH-1 (hDDAH-1) transgenicity protects from ischemic tissue damage in temporary middle cerebral artery occlusion (tMCAO) in mice. Infarct sizes did not significantly differ between hDDAH-1 transgenic (TG) mice and wild-type littermates (WT). As expected, ADMA plasma concentrations were significantly decreased, cerebral hDDAH expression and protein significantly increased in transgenic animals. Interestingly, neither brain tissue DDAH activity nor ADMA concentrations were different between TG and WT mice. In contrast, muscular DDAH activity was generally lower than in brain but significantly increased in TG mice. CONCLUSION/SIGNIFICANCE: Our study demonstrates that hDDAH-1 transgenic mice are not protected from ischemic cerebral tissue damage in tMCAO. This lack of protection is due to high basal cerebral DDAH activity, which is not further increasable by transgenic overexpression of DDAH. PMID- 19809509 TI - In vivo and ex vivo evaluation of L-type calcium channel blockers on acid beta glucosidase in Gaucher disease mouse models. AB - Gaucher disease is a lysosomal storage disease caused by mutations in acid beta glucosidase (GCase) leading to defective hydrolysis and accumulation of its substrates. Two L-type calcium channel (LTCC) blockers-verapamil and diltiazem have been reported to modulate endoplasmic reticulum (ER) folding, trafficking, and activity of GCase in human Gaucher disease fibroblasts. Similarly, these LTCC blockers were tested with cultured skin fibroblasts from homozygous point-mutated GCase mice (V394L, D409H, D409V, and N370S) with the effect of enhancing of GCase activity. Correspondingly, diltiazem increased GCase protein and facilitated GCase trafficking to the lysosomes of these cells. The in vivo effects of diltiazem on GCase were evaluated in mice homozygous wild-type (WT), V394L and D409H. In D409H homozygotes diltiazem (10 mg/kg/d via drinking water or 50-200 mg/kg/d intraperitoneally) had minor effects on increasing GCase activity in brain and liver (1.2-fold). Diltiazem treatment (10 mg/kg/d) had essentially no effect on WT and V394L GCase protein or activity levels (<1.2-fold) in liver. These results show that LTCC blockers had the ex vivo effects of increasing GCase activity and protein in the mouse fibroblasts, but these effects did not translate into similar changes in vivo even at very high drug doses. PMID- 19809510 TI - Viral load levels measured at set-point have risen over the last decade of the HIV epidemic in the Netherlands. AB - BACKGROUND: HIV-1 RNA plasma concentration at viral set-point is associated not only with disease outcome but also with the transmission dynamics of HIV-1. We investigated whether plasma HIV-1 RNA concentration and CD4 cell count at viral set-point have changed over time in the HIV epidemic in the Netherlands. METHODOLOGY/PRINCIPAL FINDINGS: We selected 906 therapy-naive patients with at least one plasma HIV-1 RNA concentration measured 9 to 27 months after estimated seroconversion. Changes in HIV-1 RNA and CD4 cell count at viral set-point over time were analysed using linear regression models. The ATHENA national observational cohort contributed all patients who seroconverted in or after 1996; the Amsterdam Cohort Studies (ACS) contributed seroconverters before 1996. The mean of the first HIV-1 RNA concentration measured 9-27 months after seroconversion was 4.30 log(10) copies/ml (95% CI 4.17-4.42) for seroconverters from 1984 through 1995 (n = 163); 4.27 (4.16-4.37) for seroconverters 1996-2002 (n = 232), and 4.59 (4.52-4.66) for seroconverters 2003-2007 (n = 511). Compared to patients seroconverting between 2003-2007, the adjusted mean HIV-1 RNA concentration at set-point was 0.28 log(10) copies/ml (95% CI 0.16-0.40; p<0.0001) and 0.26 (0.11-0.41; p = 0.0006) lower for those seroconverting between 1996-2002 and 1984-1995, respectively. Results were robust regardless of type of HIV-1 RNA assay, HIV-1 subtype, and interval between measurement and seroconversion. CD4 cell count at viral set-point declined over calendar time at approximately 5 cells/mm(3)/year. CONCLUSION: The HIV-1 RNA plasma concentration at viral set-point has increased over the last decade of the HIV epidemic in the Netherlands. This is accompanied by a decreasing CD4 cell count over the period 1984-2007 and may have implications for both the course of the HIV infection and the epidemic. PMID- 19809511 TI - The impact of nature experience on willingness to support conservation. AB - We hypothesized that willingness to financially support conservation depends on one's experience with nature. In order to test this hypothesis, we used a novel time-lagged correlation analysis to look at times series data concerning nature participation, and evaluate its relationship with future conservation support (measured as contributions to conservation NGOs). Our results suggest that the type and timing of nature experience may determine future conservation investment. Time spent hiking or backpacking is correlated with increased conservation contributions 11-12 years later. On the other hand, contributions are negatively correlated with past time spent on activities such as public lands visitation or fishing. Our results suggest that each hiker or backpacker translates to $200-$300 annually in future NGO contributions. We project that the recent decline in popularity of hiking and backpacking will negatively impact conservation NGO contributions from approximately 2010-2011 through at least 2018. PMID- 19809512 TI - Fine structures of 8-G-1-(p-YC6H4C identical with CSe)C10H6 (G = H, Cl, and Br) in crystals and solutions: ethynyl influence and Y- and G-dependences. AB - Fine structures of 8-G-1-(p-YC(6)H(4)C identical with CSe)C(10)H(6) [1 (G = H) and 2 (G = Cl): Y = H (a), OMe (b), Me (c), F (d), Cl (e), CN (f), and NO(2) (g)] are determined by the X-ray analysis. Structures of 1, 2, and 3 (G = Br) are called A if each Se-C(sp) bond is perpendicular to the naphthyl plane, whereas they are B when the bond is placed on the plane. Structures are observed as A for 1a-c bearing Y of nonacceptors, whereas they are B for 1e-g with Y of strong acceptors. The change in the structures of 1e-g versus those of 1a-c is called Y dependence in 1. The Y-dependence is very specific in 1 relative to 1-(p YC(6)H(4)Se)C(10)H(7) (4) due to the ethynyl group: the Y-dependence in 1 is almost inverse to the case of 4 due to the ethynyl group. We call the specific effect "Ethynyl Influence." Structures of 2 are observed as B: the A-type structure of 1b changes dramatically to B of 2b by G = Cl at the 8-position, which is called G-dependence. The structures of 2 and 3 are examined in solutions based on the NMR parameters. PMID- 19809513 TI - Consistent recovery of sensory stimuli encoded with MIMO neural circuits. AB - We consider the problem of reconstructing finite energy stimuli encoded with a population of spiking leaky integrate-and-fire neurons. The reconstructed signal satisfies a consistency condition: when passed through the same neuron, it triggers the same spike train as the original stimulus. The recovered stimulus has to also minimize a quadratic smoothness optimality criterion. We formulate the reconstruction as a spline interpolation problem for scalar as well as vector valued stimuli and show that the recovery has a unique solution. We provide explicit reconstruction algorithms for stimuli encoded with single as well as a population of integrate-and-fire neurons. We demonstrate how our reconstruction algorithms can be applied to stimuli encoded with ON-OFF neural circuits with feedback. Finally, we extend the formalism to multi-input multi-output neural circuits and demonstrate that vector-valued finite energy signals can be efficiently encoded by a neural population provided that its size is beyond a threshold value. Examples are given that demonstrate the potential applications of our methodology to systems neuroscience and neuromorphic engineering. PMID- 19809514 TI - Classification of nonenzymatic homologues of protein kinases. AB - Protein Kinase-Like Non-kinases (PKLNKs), which are closely related to protein kinases, lack the crucial catalytic aspartate in the catalytic loop, and hence cannot function as protein kinase, have been analysed. Using various sensitive sequence analysis methods, we have recognized 82 PKLNKs from four higher eukaryotic organisms, namely, Homo sapiens, Mus musculus, Rattus norvegicus, and Drosophila melanogaster. On the basis of their domain combination and function, PKLNKs have been classified mainly into four categories: (1) Ligand binding PKLNKs, (2) PKLNKs with extracellular protein-protein interaction domain, (3) PKLNKs involved in dimerization, and (4) PKLNKs with cytoplasmic protein-protein interaction module. While members of the first two classes of PKLNKs have transmembrane domain tethered to the PKLNK domain, members of the other two classes of PKLNKs are cytoplasmic in nature. The current classification scheme hopes to provide a convenient framework to classify the PKLNKs from other eukaryotes which would be helpful in deciphering their roles in cellular processes. PMID- 19809515 TI - Functional implication of Dp71 in osmoregulation and vascular permeability of the retina. AB - Functional alterations of Muller cells, the principal glia of the retina, are an early hallmark of most retina diseases and contribute to their further progression. The molecular mechanisms of these reactive Muller cell alterations, resulting in disturbed retinal homeostasis, remain largely unknown. Here we show that experimental detachment of mouse retina induces mislocation of the inwardly rectifying potassium channels (Kir4.1) and a downregulation of the water channel protein (AQP4) in Muller cells. These alterations are associated with a strong decrease of Dp71, a cytoskeleton protein responsible for the localization and the clustering of Kir4.1 and AQP4. Partial (in detached retinas) or total depletion of Dp71 in Muller cells (in Dp71-null mice) impairs the capability of volume regulation of Muller cells under osmotic stress. The abnormal swelling of Muller cells In Dp71-null mice involves the action of inflammatory mediators. Moreover, we investigated whether the alterations in Muller cells of Dp71-null mice may interfere with their regulatory effect on the blood-retina barrier. In the absence of Dp71, the retinal vascular permeability was increased as compared to the controls. Our results reveal that Dp71 is crucially implicated in the maintenance of potassium homeostasis, in transmembraneous water transport, and in the Muller cell-mediated regulation of retinal vascular permeability. Furthermore, our data provide novel insights into the mechanisms of retinal homeostasis provided by Muller cells under normal and pathological conditions. PMID- 19809516 TI - GLI3 repressor controls nephron number via regulation of Wnt11 and Ret in ureteric tip cells. AB - Truncating GLI3 mutations in Pallister-Hall Syndrome with renal malformation suggests a requirement for Hedgehog signaling during renal development. HH dependent signaling increases levels of GLI transcriptional activators and decreases processing of GLI3 to a shorter transcriptional repressor. Previously, we showed that Shh-deficiency interrupts early inductive events during renal development in a manner dependent on GLI3 repressor. Here we identify a novel function for GLI3 repressor in controlling nephron number. During renal morphogenesis, HH signaling activity, assayed by expression of Ptc1-lacZ, is localized to ureteric cells of the medulla, but is undetectable in the cortex. Targeted inactivation of Smo, the HH effector, in the ureteric cell lineage causes no detectable abnormality in renal morphogenesis. The functional significance of absent HH signaling activity in cortical ureteric cells was determined by targeted deletion of Ptc1, the SMO inhibitor, in the ureteric cell lineage. Ptc1(-/-UB) mice demonstrate ectopic Ptc1-lacZ expression in ureteric branch tips and renal hypoplasia characterized by reduced kidney size and a paucity of mature and intermediate nephrogenic structures. Ureteric tip cells are remarkable for abnormal morphology and impaired expression of Ret and Wnt11, markers of tip cell differentiation. A finding of renal hypoplasia in Gli3(-/-) mice suggests a pathogenic role for reduced GLI3 repressor in the Ptc1(-/-UB) mice. Indeed, constitutive expression of GLI3 repressor via the Gli3(Delta699) allele in Ptc1(-/-UB) mice restores the normal pattern of HH signaling, and expression of Ret and Wnt11 and rescued the renal phenotype. Thus, GLI3 repressor controls nephron number by regulating ureteric tip cell expression of Wnt11 and Ret. PMID- 19809518 TI - Characterization for binding complex formation with site-directly immobilized antibodies enhancing detection capability of cardiac troponin I. AB - The enhanced analytical performances of immunoassays that employed site-directly immobilized antibodies as the capture binders have been functionally characterized in terms of antigen-antibody complex formation on solid surfaces. Three antibody species specific to cardiac troponin I, immunoglobulin G (IgG), Fab, and F(ab')(2) were site-directly biotinylated within the hinge region and then immobilized via a streptavidin-biotin linkage. The new binders were more efficient capture antibodies in the immunoassays compared to randomly bound IgG, particularly, in the low antibody density range. The observed improvements could have resulted from controlled molecular orientation and also from flexibility, offering conditions suitable for binding complex formations. PMID- 19809519 TI - Postpartum ovarian vein thrombosis: two cases and review of literature. AB - Introduction. We presented two cases of late presentation of ovarian vein thrombosis postpartum following vaginal delivery and cesarean section within a short period in our institution. Both of them had pelvic pain following their deliveries which was associated with fever and chills. One of them was quite a big-sized thrombophlebitic vein which was about 10 x 6 x 5 centimeters following a computed tomography. They were both treated initially for urinary tract infection, while a large ovarian vein thrombosis was not diagnosed in the second patient until her emergency department admission. Conclusion. Ovarian vein thrombosis is rare, but could present late, and difficult to diagnose, hence, should be considered as a differential diagnosis in a postpartum woman with fever and tender pelvic mass. PMID- 19809517 TI - Synapse plasticity in motor, sensory, and limbo-prefrontal cortex areas as measured by degrading axon terminals in an environment model of gerbils (Meriones unguiculatus). AB - Still little is known about naturally occurring synaptogenesis in the adult neocortex and related impacts of epigenetic influences. We therefore investigated (pre)synaptic plasticity in various cortices of adult rodents, visualized by secondary lysosome accumulations (LA) in remodeling axon terminals. Twenty-two male gerbils from either enriched (ER) or impoverished rearing (IR) were used for quantification of silver-stained LA. ER-animals showed rather low LA densities in most primary fields, whereas barrel and secondary/associative cortices exhibited higher densities and layer-specific differences. In IR-animals, these differences were evened out or even inverted. Basic plastic capacities might be linked with remodeling of local intrinsic circuits in the context of cortical map adaptation in both IR- and ER-animals. Frequently described disturbances due to IR in multiple corticocortical and extracortical afferent systems, including the mesocortical dopamine projection, might have led to maladaptations in the plastic capacities of prefronto-limbic areas, as indicated by different LA densities in IR- compared with ER-animals. PMID- 19809520 TI - Modular Cre/lox system and genetic therapeutics for colorectal cancer. AB - The Cre/lox system is a powerful tool for targeting therapeutic effectors in a wide variety of human disorders. I review a Cre/lox Wnt-targeted system that has shown promise against Wnt-positive colorectal cancer cell lines. In addition to Wnt-specific targeting of cell death inducers, the modular nature of this gene therapy model system can be exploited by designing positive and negative feedback loops to either amplify or inhibit Wnt activity for experimental or therapeutic benefit. I discuss the structural components and performance parameters of the system, the implication of these findings with respect to cancer stem cells, as well as the general applicability of this system to any disorder characterized by differential gene expression. I also consider the issue of gene delivery as well as in vivo testing requirements necessary for the further characterization and development of this system. PMID- 19809521 TI - Efficient identification of assembly neurons within massively parallel spike trains. AB - The chance of detecting assembly activity is expected to increase if the spiking activities of large numbers of neurons are recorded simultaneously. Although such massively parallel recordings are now becoming available, methods able to analyze such data for spike correlation are still rare, as a combinatorial explosion often makes it infeasible to extend methods developed for smaller data sets. By evaluating pattern complexity distributions the existence of correlated groups can be detected, but their member neurons cannot be identified. In this contribution, we present approaches to actually identify the individual neurons involved in assemblies. Our results may complement other methods and also provide a way to reduce data sets to the "relevant" neurons, thus allowing us to carry out a refined analysis of the detailed correlation structure due to reduced computation time. PMID- 19809522 TI - FAMILY AND HUMAN RESOURCES IN THE DEVELOPMENT OF A FEMALE CRACK-SELLER CAREER: CASE STUDY OF A HIDDEN POPULATION. AB - This paper is primarily concerned with resources which family and kin network bring to drug careers. The general thesis is that specific human resources available during childhood influence both the nature and extent of participation in crack use and sales. The availability of family and human resources are critical in determining the extent to which drug abusers could develop and maintain a "conventional" identity while engaging in a drug-distribution career. Although females are becoming more evident in crack-distribution roles, they remain a minority among crack sellers and usually perform the lowest roles. This case study of Rachel represents a truly hidden population, a minority female who has been a successful crack seller for several years. The paper provides insight about persons that rarely come to attention when studying drug distribution and participation. Such persons acquire skills and resources during their lives that enable them to function in two diverse worlds. Such crack sellers are "truly hidden" because they do not have criminal records, almost never come to the attention of police, and function adequately in conventional roles. By analyzing a detailed case study of a female drug seller, this paper delineates some of the human resources and skills which may account for her differential outcome in a career of drug use and sales in inner-city settings. PMID- 19809524 TI - Qualitative Data Analysis with Hypertext: A Case of New York City Crack Dealers. AB - Ethnography has become a useful method in procuring sensitive information from the 'hidden population' who may not be accessed with quantitative survey techniques. Researchers are generating huge amounts of qualitative/textual data. Qualitative data require careful planning in storage, coding, retrieval, and analysis. Personal computers have solved data management problems, but data analysis remains problematic. The paper describes some qualitative data management and analytic problems faced by a team of ethnographers engaged in a longitudinal epidemiological study of cocaine and crack distribution/abuse in New York City. Ethnographic data was collected through multi-session open-ended interviews with more than one hundred cocaine/crack dealers and extensive field notes were kept. Compared to other programs, a hypertext software - Folio Views - was more useful in solving (a) data management and (b) analytical problems. Authors used this software to handle more than twenty-five thousand pages of texts; search and sort the database by any words or codes; and retrieve relevant textual materials needed to complete comparative and thematic analysis. Authors analyzed the data from outsiders' point of view (etic) as well as from the viewpoint of the subject populations (emic). PMID- 19809523 TI - High incidence of MGMT and RARbeta promoter methylation in primary glioblastomas: association with histopathological characteristics, inflammatory mediators and clinical outcome. AB - Glioblastomas, the most frequent primary brain tumors in adults, are characterized by a highly aggressive, inflammatory and angiogenic phenotype. Methylation of CpG islands in cancer-related genes may serve as an epigenetic biomarker for glioblastoma diagnosis and prognosis. The aim of this study was to analyze the methylation status of four critical tumor-associated genes (MGMT, RARbeta, RASSF1A, CDH13), and investigate possible links with inflammatory (interleukin [IL]-6, IL-8) and angiogenic mediators (vascular endothelial growth factor [VEGF], cyclooxygenase [COX]-2) and clinical outcome in 23 glioma samples (6 grade II astrocytomas, 17 grade IV glioblastomas). RARbeta and MGMT genes were more frequently methylated in 70.58% and 58.8% of glioblastomas, respectively. RASSF1A and CDH13 displayed a similar methylation frequency (23.52%) in glioblastomas. No gene methylation was observed in grade II astrocytomas. Tumor grade correlated positively with MGMT and RARbeta methylation (P = 0.005 and P = 0.019, respectively) and the extent of necrosis (P = 0.001 and P = 0.003). Interestingly, the marker of chronic inflammation, IL-6, was positively associated with methylation of MGMT (P = 0.004), RARbeta (P = 0.002), and RASSF1A (P = 0.0081) as well as the total number of methylated genes (P < 0.0001), indicating the important role of IL-6 in maintaining promoter methylation of these genes. VEGF expression correlated positively with MGMT and RARbeta methylation although these relationships were of marginal significance (P = 0.0679 and P = 0.0757). Kaplan-Meier univariate survival analysis indicated an unfavorable survival period in patients with MGMT methylation compared with those without methylation (P = 0.0474). Our study highlights the implication of MGMT and RARbeta methylation in the aggressive phenotype of primary glioblastomas. The association of MGMT methylation with clinical outcome indicates its potential prognostic value. PMID- 19809525 TI - Personal Safety in Dangerous Places. AB - Personal safety during fieldwork is seldom addressed directly in the literature. Drawing from many prior years of ethnographic research and from field experience while studying crack distributors in New York City, the authors provide a variety of strategies by which ethnographic research can be safely conducted in dangerous settings. By projecting an appropriate demeanor, ethnographers can seek others for protector and locator roles, routinely create a safety zone in the field, and establish compatible field roles with potential subjects. The article also provides strategies for avoiding or handling sexual approaches, common law crimes, fights, drive-by shootings, and contacts with the police. When integrated with other standard qualitative methods, ethnographic strategies help to ensure that no physical harm comes to the field-worker and other staff members. Moreover, the presence of researchers may actually reduce (and not increase) potential and actual violence among crack distributors/abusers or others present in the field setting. PMID- 19809526 TI - Gaining Access to Hidden Populations: Strategies for Gaining Cooperation of Drug Sellers/Dealers and Their Families in Ethnographic Research. AB - This article examines strategies for gaining the cooperation of drug sellers and their families in order to conduct ethnographic research. The strategies were developed during an eight year study of drug dealers in New York City. A key element in gaining the ability to talk with and observe drug dealers and their family members was the availability of funds to compensate respondents for interviews and other expenses associated with building and maintaining rapport. Access to more successful crack sellers and dealers rested upon the right contacts. The "right contact" is a critical element.Locating a trusted "go between" was adapted from strategies employed by cocaine sellers to arrange transactions involving large quantities of drugs. Such transactions rely upon a trusted associate of a dealer, the "go-between," who performs various roles and assumes risks the dealer wishes to avoid. The role of the go-between became important when ethnographers attempted to reach drug dealers for research purposes.Favors and trust are central components in the equation of access to the dealer and his family. Favors are a part of drug dealers' interaction patterns: everyone owes someone else a favor. Such reciprocity norms exist independently of the amount of drugs involved and outlast any particular transaction. Reputations and favors are related. This framework of favors, trust, and reciprocity provides a basis for the ethnographer to gain an introduction to dealers and sellers. The "go-between" is critical because he/she explains the ethnographer's role to the dealer and helps arrange an initial meeting between the ethnographer and the seller. Once the go-between has provided an initial introduction, the ethnographer marshals the communication skills necessary to convince the dealer to allow further contact and conversations.This article examines the ritual of initial conversation within its cultural framework. Developing rapport requires showing respect and honesty. Since drug dealers' self-esteem and prestige is generally tied to their drug dealing activities, signs of respect are critical in obtaining repeated appointments and conversations. Issues such as levels of rejection and how to use apparent refusal to the ethnographer's advantage are discussed. Gaining access was broken into two components. One involved permission to engage dealers in in-depth interviews The next involved obtaining permission to directly observe the actual activities of selling. Both of these components were important elements in gaining access and permission to conduct research. Building and maintaining trust and rapport were related to issues of confidentiality and anonymity.Ill-fated ethnographic strategies, such as relying on street drug users for introductions, were important stepping stones to those strategies that did work. Such strategies revealed the level of interaction between dealer and user. They helped to uncover drug subculture behavior patterns and conduct norms and to tease out the relationship between the dealer and user. Such strategies also revealed hierarchical arrangements and the loyalty within such levels. Those near the top of dealer hierarchies generally are reluctant to introduce their boss (those above them in rank) because of fear of reprisals, a sense of responsibility to the individual boss, or/and a sense of loyalty to the organization. The strategies laid out were experienced in New York and may be adjusted to acquire access to hidden populations in other situations. PMID- 19809527 TI - Mechanisms by Which Childhood Personality Traits Influence Adult Well-being. AB - Children's personality traits have enduring effects that shape adult well-being. In particular, childhood conscientiousness influences core aspects of adult well being: health, friendships, and mastery. Research is now examining the mechanisms by which early personality traits initiate and sustain particular life paths. These include mediating and moderating mechanisms that may operate during critical developmental periods or may build cumulatively over time. Future research would benefit from testing theoretically derived mechanisms for different traits, and examining variables as they change over time, using both short- and long-term longitudinal designs over different life stages. PMID- 19809528 TI - Using TIRF microscopy to quantify and confirm efficient mass transfer at the substrate surface of the chemistrode. AB - This paper describes experiments for characterizing mass transfer at the hydrophilic surface of the substrate in a chemistrode. The chemistrode uses microfluidic plugs to deliver pulses of chemicals to a substrate with high temporal resolution, which requires efficient mass transfer between the wetting layer and the hydrophilic surface of the substrate. Here, total internal reflection fluorescence microscopy (TIRFM) was used to image the hydrophilic surface of the substrate as plugs were made to flow over it. The surface of the substrate was rapidly saturated with a fluorescent dye as the fluroesecent plugs passed over the substrate, confirming effective mass transfer between the wetting layer and the surface of the substrate. The dynamics of saturation are consistent from cycle to cycle, indicating that the chemistrode can stimulate surfaces with high reproducibility. The number of plugs required to reach 90% saturation of the hydrophilic surface of the substrate, phi(90%), only weakly depended on experimental conditions (the Peclet number or the capillary number). Furthermore, over a wide range of operating conditions, phi(90%) was less than 4. These results are useful for improving the chemistrode and for understanding other phenomena that involve diffusional transfer in multiphase or recirculating flows near surfaces. PMID- 19809529 TI - Vertical migration, feeding and colouration in the mesopelagic shrimp Sergestes arcticus. AB - Intraspecific variation in vertical distribution, timing of vertical migration, and colouration of the mesopelagic shrimp Sergestes arcticus were studied in the >400 m deep part of Masfjorden, Norway. Very few individuals were caught in the upper strata during daytime, and larger individuals occurred deeper during the day than smaller ones. Vertical migration was prominent and no overall trend of increasing length with depth was found at night. Small individuals arrived in the upper layers earlier than larger ones. Animal colouration assessed by digital photography revealed significant variance in individual redness. Depth of capture was the most important factor explaining colouration, with increasing degree of redness with depth. Assessing the gut fullness of the transparent shrimps provided a rapid way of estimating feeding activity and showed that feeding took place mainly at night. PMID- 19809530 TI - In vivo MRI-Based 3D FSI RV/LV Models for Human Right Ventricle and Patch Design for Potential Computer-Aided Surgery Optimization. AB - Right ventricular dysfunction is one of the more common causes of heart failure in patients with congenital heart defects. Use of computer-assisted procedures is becoming more popular in clinical decision making process and computer-aided surgeries. A 3D in vivo MRI-based RV/LV combination model with fluid-structure interaction (FSI), RV-LV interaction, and RV-patch interaction was introduced to perform mechanical analysis for human right ventricle with potential clinical applications. Patient-specific RV/LV morphologies were acquired by using planar tagged MRI. The 3D RV/LV FSI model was solved using a commercial finite element package ADINA. Our results indicated that flow and stress/strain distributions in the right ventricle are closely related to RV morphology, material properties and blood pressure conditions. Patches with material properties better matching RV tissue properties and smaller size lead to better RV function recoveries. Computational RV volumes showed very good agreement with MRI data (error < 3%). More patient studies are needed to establish baseline database so that computational simulations can be used to replace empirical and often risky clinical experimentation to examine the efficiency and suitability of various reconstructive procedures in diseased hearts and optimal design can be found. PMID- 19809531 TI - Involvement of Youths with Autism Spectrum Disorders or Intellectual Disabilities in Multiple Public Service Systems. AB - The objectives of this study were to estimate the prevalence of autism spectrum disorders (ASD) and intellectual disability (ID) among youths active in at least one of five public service systems - mental health [MH], educational services for youth with serious emotional disturbance [SED], child welfare [CW], juvenile justice [JJ], and alcohol and drug services [AD].This study also reports the characteristics and patterns of system involvement among these youths. Results indicate that approximately 12% of a random sample of youths involved in these public service systems had ID or ASD. These disabilities were particularly prevalent in youth in the SED (25%), MH (13%), and CW (13%) systems and were less prevalent in the JJ and AD systems (4% each). Youths with ID or ASD were more likely than other youths to be Caucasian, have a higher socioeconomic status, and be more likely to have externalizing psychiatric and other problems. Of those with ASD or ID, approximately one third were served in more than one service system, with the MH and SED systems most likely to be serving youths with externalizing psychiatric disorders. These findings have important implications for service provision, treatment planning, and workforce development. PMID- 19809532 TI - "Fit" inside the Work-Family Black Box: An Ecology of the Life Course, Cycles of Control Reframing. AB - Scholars have not fully theorized the multifaceted, interdependent dimensions within the work-family "black box." Taking an ecology of the life course approach, we theorize common work-family and adequacy constructs as capturing different components of employees' cognitive appraisals of fit between their demands and resources at the interface between home and work. Employees' appraisals of their work-family linkages and of their relative resource adequacy are not made independently but, rather, co-occur as identifiable constellations of fit. The life course approach hypothesizes that shifts in objective demands/ resources at work and at home over the life course result in employees experiencing cycles of control, that is, corresponding shifts in their cognitive assessments of fit. We further theorize patterned appraisals of fit are key mediators between objective work-family conditions and employees' health, well being and strategic adaptations. As a case example, we examine whether employees' assessments on ten dimensions cluster together as patterned fit constellations, using data from a middle-class sample of 753 employees working at Best Buy's corporate headquarters. We find no single linear construct of fit that captures the complexity within the work-family black box. Instead, respondents experience six distinctive constellations of fit: one optimal, two poor, and three moderate fit constellations. PMID- 19809533 TI - The Protective Effect of Adult Mental Health Upon the Utilization of Racial Socialization Parenting Practices. AB - The purpose of this paper is to examine the impact of parental mental health and types of racial socialization. The sample consisted of 169 African-American parents and their children (ages 9-11) who participated in a federally funded research project, "Knowledge about the African American Research Experience" (KAARE). Bivariate analyses revealed significant positive relationships between parent mental health status and two forms of racial socialization: spiritual/religious coping, and racial awareness teaching, while multivariate analyses supported the positive association between parental mental health and spiritual/religious coping. These results suggest that parental characteristics may influence the use of specific types of racial socialization to assist youth in coping with discriminatory societal messages. PMID- 19809535 TI - The Role of Racial Socialization in Relation to Parenting Practices and Youth Behavior: An Exploratory Analysis. AB - Racial socialization is receiving research attention because of growing evidence that it can be a protective developmental process in African American families. The present study was an exploration of the relationship of parental mental health, discipline effectiveness, monitoring and racial socialization strategies on child externalizing behaviors in a sample of 140 African American parent/caregivers. Findings indicated that certain types of racial socialization particularly, spirituality and religious coping-in conjunction with discipline effectiveness was related to child behavior problems. Specifically, among parents who felt they used more effective discipline strategies, moderate to high rates of spiritual and religious coping were associated with a reduction of child behavior problems. These findings support the hypothesis that racial socialization is an important aspect of parenting in African American families that can be associated with the effective management of children's behavior. Implications for parenting interventions and future research are discussed. PMID- 19809536 TI - Plasma enhanced chemical vapour deposition of silica onto Ti: Analysis of surface chemistry, morphology and functional hydroxyl groups. AB - Previously, we have developed and characterised a procedure for the deposition of thin silica films by a plasma enhanced chemical vapour deposition (PECVD) procedure using tetraethoxysilane (TEOS) as the main precursor. We have used the silica coatings for improving the corrosion resistance of metals and for enhancing the bioactivity of biomedical metallic implants. Recently, we have been fine-tuning the PECVD method for producing high quality and reproducible PECVD silica (PECVD-Si) coatings on metals, primarily for biomaterial applications. In order to understand the interaction of the PECVD-Si coatings with biological species (such as proteins and cells), it is important to first analyse the properties of the silica films deposited using the optimised parameters. Therefore, this current investigation was carried out to analyse the characteristic features of PECVD-Si deposited on Ti substrates (PECVD-Si-Ti). We determined that the PECVD-Si coatings on Ti were conformal to the substrate surface, strongly adhered to the underlying substrate and were resistant to delamination. The PECVD-Si surface was composed of stoichiometric SiO(2), showed a low carbon content (below 10 at.%) and was very hydrophilic (contact angle <10 degrees ). Finally, we also showed that the PECVD-Si coatings contain functional hydroxyl groups. PMID- 19809534 TI - A Cognitive Neuroscience View of Schizophrenic Symptoms: Abnormal Activation of a System for Social Perception and Communication. AB - We will review converging evidence that language related symptoms of the schizophrenic syndrome such as auditory verbal hallucinations arise at least in part from processing abnormalities in posterior language regions. These language regions are either adjacent to or overlapping with regions in the (posterior) temporal cortex and temporo-parietal occipital junction that are part of a system for processing social cognition, emotion, and self representation or agency. The inferior parietal and posterior superior temporal regions contain multi-modal representational systems that may also provide rapid feedback and feed-forward activation to unimodal regions such as auditory cortex. We propose that the over activation of these regions could not only result in erroneous activation of semantic and speech (auditory word) representations, resulting in thought disorder and voice hallucinations, but could also result in many of the other symptoms of schizophrenia. These regions are also part of the so-called "default network", a network of regions that are normally active; and their activity is also correlated with activity within the hippocampal system. PMID- 19809538 TI - Inflammatory Mechanisms Associated with Prostatic Inflammation and Lower Urinary Tract Symptoms. AB - Inflammation is a common finding in histologic prostate specimens obtained from aging men, and accumulating data suggest that inflammation may play an important role in the development of benign prostatic hyperplasia (BPH), and the development and progression of lower urinary tract symptoms (LUTS). Inflammatory processes may contribute to prostatic enlargement directly through stimulation of prostate growth, or, alternatively, through decreasing prostatic apoptosis. Additionally, inflammatory processes may impact other components of the urogenital tract, such as the bladder, and contribute to the LUTS that may be experienced both in the presence and in the absence of prostate enlargement. Current research therefore offers clues about converging inflammatory pathways which may be targeted to improve treatment of BPH and/or LUTS as well as identifying potential targets for prevention of these syndromes. PMID- 19809537 TI - The Paradox of Oestradiol-Induced Breast Cancer Cell Growth and Apoptosis. AB - High dose oestrogen therapy was used as a treatment for postmenopausal patients with breast cancer from the 1950s until the introduction of the safer antioestrogen, tamoxifen in the 1970s. The anti-tumour mechanism of high dose oestrogen therapy remained unknown. There was no enthusiasm to study these signal transduction pathways as oestrogen therapy has almost completely been eliminated from the treatment paradigm. Current use of tamoxifen and the aromatase inhibitors seek to create oestrogen deprivation that prevents the growth of oestrogen stimulated oestrogen receptor (ER) positive breast cancer cells. However, acquired resistance to antihormonal therapy does occur, but it is through investigation of laboratory models that a vulnerability of the cancer cell has been discovered and is being investigated to provide new opportunities in therapy with the potential for discovering new cancer-specific apoptotic drugs. Laboratory models of resistance to raloxifene and tamoxifen, the selective oestrogen receptor modulators (SERMs) and aromatase inhibitors demonstrate an evolution of drug resistance so that after many years of oestrogen deprivation, the ER positive cancer cell reconfigures the survival signal transduction pathways so oestrogen now becomes an apoptotic trigger rather than a survival signal. Current efforts are evaluating the mechanisms of oestrogen-induced apoptosis and how this new biology of oestrogen action can be amplified and enhanced, thereby increasing the value of this therapeutic opportunity for the treatment of breast cancer. Several synergistic approaches to therapeutic enhancement are being advanced which involve drug combinations to impair survival signaling with the use of specific agents and to impair bcl-2 that protects the cancer cell from apoptosis. We highlight the historical understanding of oestrogen's role in cell survival and death and specifically illustrate the progress that has been made in the last five years to understand the mechanisms of oestrogen-induced apoptosis. There are opportunities to harness knowledge from this new signal transduction pathway to discover the precise mechanism of this oestrogen-induced apoptotic trigger. Indeed, the new biology of oestrogen action also has significance for understanding the physiology of bone remodeling. Thus, the pathway has a broad appeal in both physiology and cancer research. PMID- 19809539 TI - Modification in the expression of Mre11/Rad50/Nbs1 complex in low dose irradiated human lymphocytes. AB - Despite the fact that high doses of radiation are detrimental, low dose radiation (LDR) often protects the organism against a subsequent exposure of lethal doses of radiation. Present study was undertaken to understand the role of Mre11, Rad50 and Nbs1 genes in the low dose radio-adapted human peripheral blood mononuclear cells (PBMCs). Optimum time interval between low dose (0.07 Gy) and high dose (5.0 Gy) of (60)Co-gamma-radiation was observed to be 5.0 hours, at which PBMCs showed maximum LDR induced resistance (RIR). At cytogenetic level, micronuclei frequency was found to be reduced in LDR pre-irradiated PBMCs subsequently exposed to high dose radiation (HDR) as compared to controls. At transcriptional level, with reference to sham-irradiated cells significantly (p< or =0.05) altered expression of Mre11, Rad50 and Nbs1 genes was observed in low dose irradiated cells. At protein level, Mre11, Rad50 and Nbs1 were enhanced significantly (p< or =0.05) in low dose pre-irradiated cells subsequently exposed to high dose of radiation as compared to only high dose irradiated cells. Transcriptional as well as translational modulation in the expression of MRN complex components upon low dose irradiation may confer its participation in repair pathways, resulting in induced resistance. PMID- 19809540 TI - The emerging low-dose therapy for advanced cancers. AB - Generally minute doses of drugs have been prescribed in biotherapies, homeopathy, immunization and vaccinations for centuries. Now the use of low doses of drugs is on the rise to combat serious diseases such as advanced cancers around the world. This new therapeutic approach to address solid tumors and other advanced diseases is a departure from the conventional use of maximum dose protocol. A small dose of the prescribed drug is frequently administered in a continuous fashion, at regular intervals, either as a standard treatment or as a maintenance therapy for a long time. However, this new treatment method lacks any standard for drug quantization, dose fractionation, repetition frequency and duration of a treatment course for an individual patient. This paper reviews literature about metronomic therapy and discusses hormesis: both phenomena occur in low dose ranges. Better mathematical models, computer simulations, process optimization and clinical trials are warranted to fully exploit the potential of low dose metronomic therapy to cure chronic and complicated diseases. New protocols to standardize metronomic dosimetry will answer the age old questions related to hormesis and homeopathy. It appears that this new low-dose metronomic therapy will have far reaching effects in curing chronic diseases throughout the world. PMID- 19809541 TI - Interpreting 'dose-response' curves using homeodynamic data: with an improved explanation for hormesis. AB - A re-interpretation of the 'dose-response' curve is given that accommodates homeostasis. The outcome, or overall effect, of toxicity is the consequence of toxicity that is moderated by homeodynamic responses. Equilibrium is achieved by a balance of opposing forces of toxic inhibition countered by a stimulatory response. A graphical model is given consisting of two linked curves (response vs concentration and effect vs concentration), which provide the basis for a re interpretation of the 'dose-response' curve. The model indicates that such relationships are non-linear with a threshold, which is due to homeodynamic responses. Subthreshold concentrations in 'dose-response' curves provide the sum of toxic inhibition minus the homeodynamic response; the response itself is unseen in serving its purpose of neutralizing perturbation. This interpretation suggests why the alpha- and beta-curves are non-linear. The beta-curve indicates adaptive overcorrection to toxicity that confers greater resistance to subsequent toxic exposure, with hormesis as an epiphenomenon. PMID- 19809542 TI - Amifostine (WR2721) confers DNA protection to in vivo cisplatin-treated murine peripheral blood leukocytes. AB - Amifostine [S-2-3-aminopropyl amino ethyl phosphorotioic acid], a modulator agent for antineoplastic drugs involved in free radicals generation has given controversial results in cisplatin treated leukocytes in vitro. We have evaluated the amifostine protection over leukocytes in vivo, using comet assay. Groups of five OF1 male mice were given one of three doses of amifostine (56, 105 and 200 mg/Kg) after a cisplatin single injection (10 mg/Kg). Serum malonyldialdehyde levels, catalase and superoxide dismutase activity were also evaluated. Amifostine showed significant DNA protection (p< 0.01) at the two lower doses evaluated. Malonyldialdehyde decreased in all amifostine treatments with respect to cisplatin while antioxidant enzyme activities remained unchanged. However, DNA migration increased with the highest amifostine dose; in fact highest dose of amifostine did no protect damage caused by cisplatin this result have implications on amifostine treatment schedules in clinical practice. PMID- 19809543 TI - Turnover modeling of non-esterified fatty acids in rats after multiple intravenous infusions of nicotinic acid. AB - The objective of this investigation was to use a pharmacokinetic (PK)/pharmacodynamic (PD) approach to describe and evaluate a PK model of nicotinic acid (NiAc) in guinea pigs and a PD feedback model of changes in non esterified fatty acid (NEFA) concentrations in rats following multiple intravenous infusions of NiAc at different rates and durations of inhouse and literature (NEFA after extravascular NiAc dosing) data. Serial arterial blood samples were taken for evaluation of NiAc exposure in guinea pigs and NEFA in rats. The biophase kinetics of NiAc was assumed to impact on NEFA turnover with feedback incorporated via an inhibitory moderator compartment. The response acted linearly on the production of moderator, which then acted inversely on the turnover rate of response. The potency, expressed as the amount of NiAc in the biophase causing a 50 % inhibitory effect (ID(50)), was 6.5 nmol +/- 31 % and the half-life of response (t(1/2, kout)) 2 min +/- 18 %. The half-life of tolerance (t(1/2, ktol)) was 9 min +/- 27 %. The model can be used to provide information about factors that determine the time course of NEFA response following different rates and routes of administration of NiAc or NiAc analogues. PMID- 19809544 TI - Interaction between short-term heat pretreatment and avermectin on 2nd instar larvae of diamondback moth, Plutella xylostella (Linn). AB - Based on the cooperative virulence index (c.f.), the interaction effect between short-term heat pretreatment and avermectin on 2(nd) instar larvae of diamondback moth (DBM), Plutella xylostella (Linnaeus), was assessed. The results suggested that the interaction results between short-term heat pretreatment and avermectin on the tested insects varied with temperature level as well as its duration and avermectin concentration. Interaction between heat pretreatment at 30 degrees C and avermectin mainly resulted in addition. Meanwhile, pretreatment at 35 degrees C for 2 or 4 h could antagonize the toxicity of avermectin at lower concentrations, which indicated a hormetic effect occurred. The results indicate that cooperative virulence index (c.f.) may be adopted in hormetic effect assessment. PMID- 19809545 TI - International Federation of Gynecology and Obstetrics (FIGO) staging system revised: what should be considered critically for gynecologic cancer? PMID- 19809546 TI - Recent advances of robotic surgery and single port laparoscopy in gynecologic oncology. AB - Two innovative approaches in minimally invasive surgery that have been introduced recently are the da Vinci robotic platform and single port laparoscopic surgery (SPLS). Robotic surgery has many advantages such as 3-dimensional view, the wrist like motion of the robotic arm and ergonomically comfortable position for the surgeon. Numerous literatures have demonstrated the feasibility of robotic surgery in gynecologic oncology. However, further research should be performed to demonstrate the superiority of robotic surgery compared to conventional laparoscopy. Additionally, cost reduction of robotic surgery is needed to adopt robotic surgery into gynecologic oncology worldwide. SPLS has several possible benefits including reduced operative complications, reduced postoperative pain, and better cosmetic results compared to conventional laparoscopy. Although several authors have indicated that SPLS is a feasible approach for gynecologic surgery, there have been few reports demonstrating the potential advantages over conventional laparoscopy. Moreover, technical difficulties of SPLS still exist. Therefore, the advantages of a single port approach compared to conventional laparoscope should be evaluated with comparative study, and further technologic development for SPLS is also needed. These two progressive technologies take the lead in the development of MIS and further studies should be performed to evaluate the benefits of robot surgery and SPLS. PMID- 19809547 TI - Prognostic significance of neutropenia during adjuvant concurrent chemoradiotherapy in early cervical cancer. AB - OBJECTIVE: To evaluate the prognostic significance of adjuvant concurrent chemoradiotherapy-induced neutropenia with survival in patients with squamous cell carcinoma of the uterine cervix. METHODS: Data from 107 patients with stage IB-IIB cervical cancer were retrospectively analyzed. The median follow-up was 37.5 (4.2-72.7) months. All patients had received radical surgery, including pelvic lymphadenectomy, followed by paclitaxel plus carboplatin-based concurrent chemoradiotherapy. Relative neutropenia, defined as an absolute neutrophil count <1,000/mm(3) at the concurrent chemoradiotherapy cycle nadir, correlated to the pathologic findings and survival outcomes. RESULTS: Sixty-six patients experienced neutropenia at least once during concurrent chemoradiotherapy, and demonstrated marginal improvement in disease-free survival (p=0.055), although not in overall survival. By subgroup analyses, the gain of disease free survival mainly originated from the node metastasis subgroup (p=0.033). Treatment-induced neutropenia proved to be the only significant independent factor for recurrence in cervical cancer (p=0.042) by multivariate analysis. CONCLUSION: Concurrent chemoradiotherapy-induced neutropenia may be a prognostic factor of recurrence in patients with cervical cancer. Individualized dose titration of the tolerable myelosuppression might be beneficial. PMID- 19809548 TI - Acute toxicity of cyclooxygenase-2 inhibitor rofecoxib as a radiosensitizer for concurrent chemoradiation in the treatment of uterine cervical cancer. AB - OBJECTIVE: To evaluate the acute toxicity of rofecoxib during concurrent use with cisplatin-based chemoradiotherapy (CCRT) in patients with cervical cancer. METHODS: We evaluated 67 FIGO stage IB2-IVA cervical cancer patients treated with CCRT between June 2002 and July 2004. The study group included patients who received rofecoxib (N=30) and the control group included patients who received CCRT only (N=37). The patients' medical records were retrospectively reviewed for patient characteristics, toxicity related to CCRT and treatment results. RESULTS: There were no significant differences in toxicity between the two groups. The most common acute grade 3/4 toxicity was neutropenia (13.3% in the study group and 21.6% in the control group). Grade 3/4 late toxicity was observed in 2 (6.6%) patients in the study group and 3 (8.1%) in the control group. There was no treatment-related deaths in either group. Six (20.0%) patients in the study group had treatment failure. In the control group, 6 (16.2%) patients experienced treatment failure. Progression-free and overall survival was 55.8+/-4.2 and 59.0+/-2.8 months, respectively, in the study group, and 69.7+/-4.3 and 71.6+/ 3.6 months, respectively, in the control group. There were no differences in progression-free and overall survival between the 2 groups. CONCLUSION: Our data indicate that rofecoxib, at a dose of 25 mg twice daily, has acceptable acute toxicity as a radiosensitizer during CCRT. Although rofecoxib was not efficacious as a radiosensitizer in the present study, the benefit of rofecoxib as a radiosensitizer should be further evaluated in a prospective study. PMID- 19809549 TI - Prognostic factors in adult granulosa cell tumors of the ovary: a retrospective analysis of 80 cases. AB - OBJECTIVE: Ovarian granulosa cell tumors are rare malignancies with a relatively favorable prognosis. However, patients still suffer from disease-related mortality. Therefore, the prognostic factors should be clarified. The purpose of this study was to investigate the clinical and pathologic characteristics related with disease recurrence and mortality in adult type ovarian granulosa cell tumors. METHODS: Eighty surgically staged patients with granulosa cell ovarian tumor treated at the Hacettepe University Hospital between 1982 and 2006 were retrospectively reviewed. Clinical and pathological characteristics were analyzed. RESULTS: Granulosa cell ovarian tumors accounted for 4.3% of malignant ovarian neoplasms. Mean age was 47.6 years. The most common presenting symptom was abnormal uterine bleeding (53.7%). Endometrial pathology was detected in 51.2% of patients preoperatively. Seventy percent of patients were diagnosed at stage I, and 53.8% of patients received adjuvant treatment. Mean follow-up was 67.5 months. Overall 5-year and 10-year survival was 91% and 86%, respectively. Mean survival was 147.1 months. Recurrence rate was 11.2%. In univariate analysis, advanced stage, advanced age, residual disease after surgery, and need for adjuvant treatment were associated with disease-related mortality and advanced stage disease and absence of initial staging surgery were associated with disease recurrence. However, in multivariate analysis, only initial stage was found to be a significant prognostic factor. CONCLUSION: Initial stage seems to be the single most important prognostic factor in ovarian granulosa cell tumors. Therefore, a comprehensive staging surgery should be attempted to document the real extent of disease and to estimate the oncologic outcome more accurately. PMID- 19809550 TI - Treatment of the patients with abnormal cervical cytology: a "see-and-treat" versus three-step strategy. AB - OBJECTIVE: To examine the correlation between cervical cytology and final histological results in patients who have undergone loop electrosurgical excision procedure (LEEP) with or without colposcopy-directed biopsy. METHODS: A retrospective review was performed of 829 patients who underwent LEEP for abnormal cervical cytology at Gangnam Severance Hospital between January 2004 and December 2008. Patients were classified to three groups according to cervical cytology and also divided into two groups based on the treatment they received: see-and-treat group and the standard three-step group. Final histological results were compared for the each study group. RESULTS: There were no differences in the final histological results between see-and-treat and three-step group in patients with high-grade squamous intraepithelial lesions (HSIL) cytology (N=523) (p=0.71). However, in patients with low-grade squamous intraepithelial lesions (LSIL)/atypical squamous cells of undetermined significance (ASCUS) (N=257) or normal cytology (N=49), the final histological results were significantly different between see-and-treat and three-step group (p<0.001) and the rate of overtreatment was significantly higher in the see-and-treat group (p<0.001). CONCLUSION: A see-and-treat protocol may be a viable alternative only in patients with HSIL cytology if colposcopic impression is suggestive of cervical intraepithelial neoplasia (CIN) 2 or 3 lesions. PMID- 19809551 TI - Correlation between preoperative serum levels of five biomarkers and relationships between these biomarkers and cancer stage in epithelial overian cancer. AB - OBJECTIVE: To examine the correlation among the preoperative serum levels of five biomarkers presumed to be useful for early detection of epithelial ovarian cancer and evaluate the relationships between serum levels of these five biomarkers and epithelial ovarian cancer stage. METHODS: We analyzed 56 newly diagnosed epithelial ovarian cancer patients. Preoperative serum levels of leptin, prolactin, osteopontin (OPN), insulin-like growth factor-II, and CA-125 were determined by ELISA. We also examined the correlation between the serum levels of the biomarkers and ovarian cancer stage. Significant differences in the mean serum levels of two proteins, leptin and CA-125, were observed between stage subsets. RESULTS: There was a significant negative correlation between prolactin and leptin and a significant positive correlation between prolactin and OPN. Of the five biomarkers, only the mean serum CA-125 level showed a significant positive correlation with cancer stage (Spearman rho=0.24, p<0.01). OPN showed a marginally significant positive correlation with stage (Spearman rho=0.14, p=0.07). CONCLUSION: We demonstrated the relationship between five biomarkers in epithelial ovarian cancer. These tumor markers may be useful in screening for ovarian cancer, in characterizing disease states, and in developing therapeutic interventions targeting these marker proteins. Large-scale studies that include potential confounding factors and modifiers are necessary to more accurately define the value of these novel biomarkers in ovarian cancer. PMID- 19809552 TI - Clinical analysis of intra-operative frozen section proven borderline tumors of the ovary. AB - OBJECTIVE: We have assessed the accuracy of frozen section diagnosis and the outcomes of misdiagnosis in borderline tumors of the ovary (BTO) according to frozen section. METHODS: All pathology reports with BTO in both frozen and permanent section analyses between 1994 and 2008 at Seoul St. Mary's Hospital were reviewed. Frozen section diagnosis and permanent section histology reports were compared. Logistic regression models were conducted to evaluate the correlation of patient and tumor characteristics with diagnostic accuracy. The clinical outcomes of misdiagnosis were evaluated. RESULTS: Agreement between frozen section diagnosis and permanent histology was observed in 63 of 101 patients (62.4%). Among the 76 patients with frozen section proven BTO, under diagnosis and over-diagnosis occurred in 8 of 76 (10.5%) and 5 of 76 patients (6.6%), respectively. Mean diameter of under-diagnosed tumor was larger than matched BTO (21.0+/-11.4 vs. 13.7+/-7.1; p=0.021). Tumor size 20 cm was determined as the optimal cut-off for under-diagnosis (50% sensitivity, 87.3% specificity). Among 8 under-diagnosed patients, no patient relapsed. Among 5 over diagnosed patients, 2 patients < 35 years of age had fertility-preserving surgery. CONCLUSION: Although frozen section diagnosis is an important and reliable tool in the clinical management of patients with ovarian tumors, over diagnosis and under-diagnosis are relatively frequent in frozen proven BTO. Surgical decision-making for BTO based on frozen section diagnosis should be done carefully, especially in large tumors. PMID- 19809553 TI - Premenopausal early-stage endometrial carcinoma patients with low CA-125 levels and low tumor grade may undergo ovary-saving surgery. AB - OBJECTIVE: The purpose of this study was to determine the possible predicting factors of coexisting adnexal malignancies, and to evaluate the safety of ovary saving surgery for early-stage endometrial carcinoma in premenopausal patients. METHODS: A retrospective review of 107 patients with endometrial carcinoma who underwent surgical treatment at our institution was conducted. All patients were younger than 50 years of age and premenopausal status. Statistical analysis was performed. RESULTS: Of the 107 patients, 78 patients had stage I to II disease and both preoperative CA-125 levels were measured and tumor grades evaluated. On multivariate analysis, preoperative CA-125 levels (p=0.018) and preoperative tumor grade (p=0.029) were independent predicting factors of adnexal diseases. The risk of coexisting ovarian malignancy was 1.8% in patients with preoperative CA-125 levels less than or equal to 34.5 U/ml and preoperative tumor grade 1 or 2. The risk increases to 20% for low CA-125 and grade 3, 13.3% for high CA-125 and grade 1 or 2, and 100% for high CA-125 and grade 3. Between patients who underwent unilateral salpingo-oophorectomy and those who underwent bilateral salpingo-oophorectomy, there was no statistically significant difference in terms of BMI, preoperative CA-125 levels, FIGO stage, histology, tumor grade, lymphadenectomy, and adjuvant treatment. CONCLUSION: Ovary-saving surgery for premenopausal, early-stage endometrial cancer patients may be considered as a treatment option in those with low preoperative CA-125 and low tumor grade. PMID- 19809554 TI - A case of neutrophilia related to a cytokine-producing relapsed squamous cell carcinoma of the uterine cervix arising from the rectovaginal septum. AB - Paraneoplastic neutrophilia caused by a squamous cell carcinoma of the uterine cervix has been seen rarely. We report a case of relapsed squamous cell carcinoma of the uterine cervix with severe neutrophilia, rapid tumor growth and aggressive clinical course, possibly due to autocrine stimulation of cell growth by G-CSF and IL-6 without other possible causes of neutrophilia. PMID- 19809555 TI - A case of non-gestational choriocarcinoma arising in the ovary of a postmenopausal woman. AB - Primary ovarian choriocarcinoma arising from a germ cell is an extremely rare occurrence, especially in postmenopausal women, and the prognosis is poor. Non gestational choriocarcinoma of the ovary (NGCO) accounts for 0.6% or less of all ovarian neoplasms. It is important to distinguish gestational choriocarcinomas of the ovary (GCO) from other carcinomas because of the poor prognosis of NGCO. We describe a case of NGCO with lung metastasis in a 55 year old woman, which we present together with a brief review of the literature. PMID- 19809556 TI - A single horn endometrial carcinoma of a uterus bicornis unicollis. AB - In this report, we describe a case of endometrial carcinoma arising in one horn of a bicornuate uterus. The diagnosis of this rare combination can be missed unless an unrecognized postmenopausal bleeding alerts the gynecologist to make a careful search for both endometrial cavities that may be curetted. Physicians should remember the possible existence of a separate uterine cavity when endometrial cancer is clinically suspected but histology fails to confirm the diagnosis. PMID- 19809557 TI - Prognostic factors of secondary cytoreductive surgery for patients with recurrent epithelial ovarian cancer. PMID- 19809558 TI - In reply: Different role of secondary cytoreductive surgery by surgeon's experience and hospital facility. PMID- 19809559 TI - Erratum: ERRATUM: Role of high risk-human papilloma virus test in the follow-up of patients who underwent conization of the cervix for cervical intraepithelial neoplasia. AB - [This corrects the article on p. 86 in vol. 20, PMID: 19590718.]. PMID- 19809560 TI - Immunomodulation of breast cancer via tumor antigen specific Th1. AB - It has long been assumed that the immune system plays a role in tumor eradication, however, scant clinical evidence exists to support that hypothesis. In recent years, as the immune system and its specific effector cells are better defined, convincing data supporting immune surveillance is emerging. Several studies have shown that an "immune signature" in the tumor microenvironment is associated with a superior outcome in a variety of cancer types. Moreover, studies have suggested that T cells found in high density within the tumor parenchyma are also correlated with a survival benefit. The type of adaptive immune response implicated in improved cancer outcomes is a type 1 response. That is, adaptive immunity associated with T cells that secrete pro-inflammatory cytokines, such as IFN-gamma, which can not only support a proliferative antigen specific T cell response but also enhance "cross priming" by activating antigen presenting cells local to the tumor site. There are many methods available that will allow the development of clinical reagents designed to stimulate Th1 immunity; either by in vitro or in vivo manipulation. Clinical trials of a variety of immunotherapeutic strategies indicate that the generation of tumor antigen specific Th1 may be beneficial in inhibiting the growth of common solid tumors. PMID- 19809561 TI - Nationwide cancer incidence in Korea, 2003-2005. AB - PURPOSE: To estimate the current cancer burden in Korea, newly diagnosed cancer cases and cancer incidence rates were calculated for the years 2003~2005. MATERIALS AND METHODS: The cancer incidence cases and rates were calculated from the Korea National Cancer Incidence Database. Crude and age-standardized incidence rates were calculated by gender for specified cancer sites in 5-year age groups. RESULTS: From 2003 to 2005, 398,824 cases of cancer were newly diagnosed in Korea (218,856 in men and 179,968 in women). For all sites combined, the crude incidence rate (CR) was 300.0 and 248.2 for men and women and the age standardized incidence rate (ASR) was 297.0 and 191.2 per 100,000, respectively. Among men, five leading cancers were stomach (CR 66.0, ASR 64.2), lung (CR 48.5, ASR 50.3), liver (CR 44.9, ASR 42.1), colon and rectum (CR 37.9, ASR 37.2), and prostate cancer (CR 12.7, ASR 13.8). Among women, five leading cancers were breast (CR 37.3, ASR 29.0), thyroid (CR 36.2, ASR 28.8), stomach (CR 34.1, ASR 25.4), colon and rectum (CR 28.0, ASR 21.1), and lung cancer (CR 17.9, ASR 12.8). In the 0~14-year-old group, leukemia was the most common in both sexes; in the 15~34 group, the most common cancer was stomach cancer for men and thyroid cancer for women; in the 35~64 group, stomach cancer for men and breast cancer for women; among those 65 and over, lung cancer for men and stomach cancer, for women, respectively. CONCLUSION: The cancer incidence rates have increased in recent years, and more cancers are expected to develop as Korea is quickly becoming an aged society. The cancer incidence statistics in this report can be used as an important source to effectively plan and evaluate the cancer control program in Korea. PMID- 19809562 TI - The role of radiotherapy in the treatment of newly diagnosed supratentorial low grade oligodendrogliomas: comparative analysis with immediate radiotherapy versus surgery alone. AB - PURPOSE: The purpose of this study was to evaluate the role of immediate postoperative radiotherapy (RT) in adult patients with a low-grade oligodendroglioma (LODG). MATERIALS AND METHODS: A total of 74 patients, older than 15 years, were treated in our institution between April 1990 and March 2006 for newly diagnosed LODGs. After surgery, 43 patients were treated with immediate RT with a total dose of 54~55.8 Gy with 1.8 Gy fractions (RT group) and 31 patients were followed with no adjuvant RT (OP group). All patients were closely observed until tumor progression or death with frequent work-ups including neurological examinations and MRI. Primary endpoints were overall survival and progression-free survival. The median follow-up duration of survivors was 6.2 years in the RT group and 5.8 years in the OP group. RESULTS: Median progression free survival was 13.2 years in the RT group and 4.6 years in the OP group; multivariate analysis confirmed improved outcome with the use of immediate RT (hazard ratio, 0.22; 95% confidence interval-CI, 0.09~0.55; p<0.001). Median overall survival was 14.9 years in the RT group and 9.8 years in the OP group; the use of adjuvant RT was also associated with a trend toward better overall survival after immediate RT based on multivariate analysis (hazard ratio, 0.3; 95% CI, 0.08~1.17; p=0.082). No severe RT related complications were observed. CONCLUSION: Immediate RT following surgery appears to be an effective treatment modality for supratentorial LODGs. However, the potential benefit of adjuvant RT for overall survival needs to be tested prospectively in the future. PMID- 19809563 TI - Clinical value of ezrin expression in primary osteosarcoma. AB - PURPOSE: Ezrin is a membrane cytoskeletal linker protein and it is known to be associated with metastasis of primary osteosarcoma. The aim of this study is to determine the relationship between an ezrin expression and several key clinical parameters and to elucidate its potential prognostic value for patients with osteosarcoma. MATERIALS AND METHODS: Seventy patients with histologically confirmed osteosarcoma and who had no distant metastasis were enrolled between 1995 and 2005 at Yonsei Cancer Center, Severance Hospital, Korea. The clinical parameters were retrospectively reviewed and immunohistochemical staining (IHC) for ezrin was performed using the surgically resected specimens. RESULTS: Of the 70 tumor specimens, 39 (55.7%) revealed an ezrin expression. More of an osteoblastic histology and an elevated initial ALP level were observed in the ezrin positive patients than in the ezrin negative patients (p=0.008 and 0.001, respectively). The proportion of patients who favorably responded to neoadjuvant chemotherapy (>=or=90% necrosis) was significantly higher in the group of ezrin positive patients than that in the group of ezrin negative patient (72.2% vs 45.2%, respectively, p=0.024). The ezrin positive patients showed more frequent recurrence than did the ezrin negative patients (64.1% vs 35.5%, respectively, p=0.017). The patients with an ezrin expression also demonstrated poorer survival than did those patients without ezrin expression (5-year EFS: 31.7% vs 61.3%, respectively, p=0.023, 5-year OS: 53.4% vs 71.0%, respectively, p=0.022). When comparing EFS according to both an ezrin expression and chemoresponsiveness, there were trends that the ezrin negative/chemoresponsive group showed the best 5 year EFS (71.4%), followed by the ezrin negative/chemoresistant group (52.9%), the ezrin positive/chemoresponsive group (38.1%) and the ezrin positive/chemoresistant group (13.6%). These trends were statistically significant (p=0.036). CONCLUSION: The expression of ezrin by IHC staining was found in 55.7% of the patients with metastasis-free osteosarcoma. Immunoreactivity to ezrin is a negative prognostic factor for survival for the patients suffering with osteosarcoma. Identifying an ezrin expression might offer a valuable piece of information when treating patients with primary osteosarcoma. PMID- 19809564 TI - Apoptosis and expression of AQP5 and TGF-beta in the irradiated rat submandibular gland. AB - PURPOSE: To evaluate the effect of X-ray irradiation on apoptosis and change of expression of aquaporin 5 (AQP5) and transforming growth factor-beta(TGF-beta) in the rat submandibular gland (SMG). MATERIALS AND METHODS: SMGs of 120 male Sprague-Dawley rats were irradiated with a single X-ray dose (3, 10, 20, or 30 Gy). At the early and late post-irradiation phase, apoptosis was measured by the terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) method, and expression of AQP5 and TGF-beta was determined by immunohistochemical staining. RESULTS: At the late post-irradiation phase, increased apoptosis was evident and marked decreases of expression of AQP5 expression by acinar cells and TGF-beta expression by ductal cells were evident. CONCLUSION: Apoptosis after X ray irradiation develops relatively late in rat SMG. Irradiation reduces AQP5 and TGF-beta expression in different SMG cell types. PMID- 19809565 TI - Leptin and leptin receptor expression in breast cancer. AB - PURPOSE: Leptin is a multifunctional hormone that's produced by adipose tissue and leptin is involved in the regulation of food intake and energy balance. The aims of this study were to determine the leptin and leptin receptor (Ob-R) expressions in human breast cancer and their corresponding influence on the prognosis of patients with breast cancer. MATERIALS AND METHODS: We examined the correlations between the leptin and Ob-R expressions and the breast cancer related pathobiologic markers by performing immunohistochemistry in 517 patients with breast cancer. We analyzed the leptin and Ob-R expressions with respect to overall survival and relapse-free survival (RFS). RESULTS: Positive cytoplasmic immunoreactivity for leptin was noted in 39% of the patients and 79% of the patients showed positive cytoplasmic immunoreactivity for Ob-R. The expression of leptin in breast cancer was correlated with a high Ki-67 labeling index (p=0.019). Based on the univariate survival analysis, the clinicopathologic variables with prognostic value included the histologic grade, the T stage, the N stage, the HER2 status, the Bcl-2, p53 and Ki-67 expressions (p<0.05). The patients with leptin-positive breast cancers and a negative hormone receptor status had a significantly longer overall survival (p=0.021). Multivariate survival analysis showed that a positive expression of leptin was an independent prognostic marker for overall survival (hazard ratio, 0.20; 95% CI, 0.04~0.99; p=0.05). CONCLUSION: A leptin expression in breast cancer is significantly associated with the Ki-67 labeling index, and this suggests there is an association of a leptin expression with the proliferation activity. In addition, a leptin expression is an indicator of better survival for breast cancer patients. PMID- 19809566 TI - TNF-alpha downregulates E-cadherin and sensitizes response to gamma-irradiation in Caco-2 cells. AB - PURPOSE: The purpose of the present study was to assess the biological effects of TNF-alpha in Caco-2 well-differentiated colon adenocarcinoma cells and to determine radiation sensitivity in order to develop TNF-alpha into a cancer therapeutic agent. MATERIALS AND METHODS: A cell viability test was conducted via a colorimetric and colony forming assay after 1 day and 3 days of incubation with TNF-alpha. Western blotting analysis and immunofluorescence staining were conducted to explore TNF-alpha-induced morphological and molecular changes in the adhesion molecules, E-cadherin and claudin-4. The effects of gamma-irradiation at a dose of 2 Gy on cell survival were evaluated by a clonogenic assay. The molecular changes in apoptosis-regulatory proteins were assessed by Western blotting. RESULTS: Caco-2 cells were highly resistant to TNF alpha-induced cell death and 2 Gy of gamma-irradiation. However, we observed the downregulation of the adherens junctional protein, E-cadherin and translocation of tight junctional protein, claudin-4 from the membrane to the cytosol induced by TNF-alpha treatment which would indicate cell-cell junction disruptions. These alterations of junctional proteins influenced the regulation of cell death in response to 2 Gy of gamma-irradiation. The combined treatment of TNF-alpha with 2 Gy of gamma irradiation reduced the survival of Caco-2 cells by down-regulating bcl-xl and activating JNK pathways. CONCLUSION: These results suggest that TNF-alpha might be potentially applied as a therapeutic agent in order to enhance sensitivity to 2 Gy of gamma-irradiation administered in radiotherapy for the treatment of human colon cancer. PMID- 19809567 TI - F-18 FDG PET-positive fibrous dysplasia in a patient with intestinal non Hodgkin's lymphoma. AB - Fibrous dysplasia (FD) is a common benign bone disorder of an unclear etiology. It is known that FD can appear without an increased FDG uptake on F-18 fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT). However, there are also several reports that FD showed increased FDG uptake and this mimicked malignant bone involvement on FDG-PET. Herein we describe a case of biopsy-proven FDG-PET positive FD in a patient with intestinal non-Hodgkin's lymphoma (NHL). A 45-year-old woman was diagnosed with intestinal NHL, which was removed by right hemicolectomy. After the operation, the FDG-PET/CT scan showed hypermetabolic activity in the right transverse process of the T10 vertebra. The patient then received a total of 6 cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone) chemotherapy every 3 weeks. After completion of the planned chemotherapy, the 2(nd) FDG-PET/CT showed increased FDG uptake (SUVmax=6.0 g/mL) of the previous bone lesion. The MR images revealed a T1 hypointense lesion with sharp borders in the same region, and this showed homogenous contrast enhancement on the fat-suppressed T1-weighted images. After the radiologic studies were carefully reviewed, the bone lesion was assumed to be benign such as FD. We performed bone biopsy and the histological examination confirmed the diagnosis of FD. In conclusion, bone lesions with FDG uptake need to be carefully interpreted when evaluating patients with known malignancy. PMID- 19809568 TI - Clear cell carcinoma of the pancreas--a case report and review of the literature. AB - Most of the malignant neoplasms of the pancreas demonstrate features that are consistent with adenocarcinoma. According to the WHO classification, primary clear cell carcinoma of the pancreas is rare and it is classified as a "miscellaneous" carcinoma. In addition, there is not an adequate systematic overview that can demonstrate its true existence as a definable entity. We report here on an unusual case of primary pancreatic clear cell carcinoma, which is the first such reported case in Korea. A 66 year old woman presented with abdominal pain and significant weight loss over the previous three weeks. On the abdominal computed tomography (CT), we detected an abdominal mass involving the pancreas tail and liver, and clear cell carcinoma with rhabdoid feature was seen on the histologic evaluation. The tumor cells showed well defined cell membranes, clear cytoplasm and prominent cell boundaries. The immunohistochemical stains showed positive reactions to antibodies against pan-cytokeratin, cytokeratin 7, carcinoembryonic antigen (CEA) and epithelial membrane antigen (EMA). On the other hand, there was a negative reaction for cytokeratin 20, chromogranin, synaptophysin, smooth muscle actin and HMB-45. She was diagnosed with a primary pancreatic clear cell carcinoma with hepatic metastasis and she received palliative gemcitabine chemotherapy. The patient died one month later of pancreatic cancer progression. PMID- 19809569 TI - Plasmablastic lymphoma in the anal canal. AB - Plasmablastic lymphoma (PBL) of the oral cavity is an acquired immunodeficiency syndrome-related lymphoma. The immunophenotype of this disease is associated with poor expression of B-cell markers but a positive reactivity for plasma cell markers. PBL is highly aggressive and responds poorly to treatment. Although originally described in the oral cavity, this disease can occur in other body niches. Here, we describe a very rare case of PBL in the anal canal of a 40-year old woman with human immunodeficiency virus infection. The malignant cells were positive for Epstein-Barr virus and human herpes virus 8. PMID- 19809570 TI - Clathrin couture: fashioning distinctive membrane coats at the cell surface. PMID- 19809571 TI - Distinct dynamics of endocytic clathrin-coated pits and coated plaques. AB - Clathrin is the scaffold of a conserved molecular machinery that has evolved to capture membrane patches, which then pinch off to become traffic carriers. These carriers are the principal vehicles of receptor-mediated endocytosis and are the major route of traffic from plasma membrane to endosomes. We report here the use of in vivo imaging data, obtained from spinning disk confocal and total internal reflection fluorescence microscopy, to distinguish between two modes of endocytic clathrin coat formation, which we designate as "coated pits" and "coated plaques." Coated pits are small, rapidly forming structures that deform the underlying membrane by progressive recruitment of clathrin, adaptors, and other regulatory proteins. They ultimately close off and bud inward to form coated vesicles. Coated plaques are longer-lived structures with larger and less sharply curved coats; their clathrin lattices do not close off, but instead move inward from the cell surface shortly before membrane fission. Local remodeling of actin filaments is essential for the formation, inward movement, and dissolution of plaques, but it is not required for normal formation and budding of coated pits in the cells we have studied. We conclude that there are at least two distinct modes of clathrin coat formation at the plasma membrane--classical coated pits and coated plaques--and that these two assemblies interact quite differently with other intracellular structures. PMID- 19809573 TI - SEMIPARAMETRIC TRANSFORMATION MODELS WITH RANDOM EFFECTS FOR CLUSTERED FAILURE TIME DATA. AB - We propose a general class of semiparametric transformation models with random effects to formulate the effects of possibly time-dependent covariates on clustered or correlated failure times. This class encompasses all commonly used transformation models, including proportional hazards and proportional odds models, and it accommodates a variety of random-effects distributions, particularly Gaussian distributions. We show that the nonparametric maximum likelihood estimators of the model parameters are consistent, asymptotically normal and asymptotically efficient. We develop the corresponding likelihood based inference procedures. Simulation studies demonstrate that the proposed methods perform well in practical situations. An illustration with a well-known diabetic retinopathy study is provided. PMID- 19809572 TI - Reading Faces for Information about Words and Emotions in Adolescents with Autism. AB - Studies of explicit processing of facial expressions by individuals with autism spectrum disorder (ASD) have found a variety of deficits and preserved abilities compared to their typically developing (TD) peers. However, little attention has been paid to their implicit processing abilities for emotional facial expressions. The question has also been raised whether preferential attention to the mouth region of a speaker's face by ASD individuals has resulted in a relative lipreading expertise. We present data on implicit processing of pseudo dynamic facial emotions and visual speech in adolescents with autism. We compared 25 ASD and 25 TD participants on their ability to recreate the sequences of four dynamic emotional facial expressions (happy, sad, disgust, fear) as well as four spoken words (with, bath, thumb, watch) using six still images taken from a video sequence. Typical adolescents were significantly better at recreating the dynamic properties of emotional expressions than those of facial speech, while the autism group showed the reverse accuracy pattern. For Experiment 2 we obscured the eye region of the stimuli and found no significant difference between the 22 adolescents with ASD and 22 TD controls. Fearful faces achieved the highest accuracy results among the emotions in both groups. PMID- 19809574 TI - The interleukin-1 receptor-associated kinase M selectively inhibits the alternative, instead of the classical NFkappaB pathway. AB - The innate immunity signaling process is controlled by numerous positive and negative regulators. The interleukin-1 receptor-associated kinase M (IRAK-M) is one of the negative regulators that contribute to the attenuation of NFkappaB activation. The molecular mechanism involved, however, is poorly defined. In this report, we observed that IRAK-M selectively suppresses the NIK-IKKalpha-mediated alternative NFkappaB pathway. Deletion of IRAK-M led to NIK stabilization, favored the formation of the IKKalpha/IKKalpha homodimer instead of the IKKalpha/IKKbeta heterodimer, and enhanced RelB nuclear distribution. In contrast, p65 nuclear localization and phosphorylation was not affected by IRAK-M deficiency. IRAK-M-deficient cells exhibited increased expression of selected cytokines such as IL-6 and GM-CSF, as well as quickened resynthesis of IkappaBalpha. The increased expression of IL-6 and GM-CSF was ablated when RelB expression was knocked down using specific siRNA. We also demonstrated that the observed inhibitory effect of IRAK-M was primarily limited to the TLR2 ligand, instead of TLR4. Taken together, our findings suggest that IRAK-M negatively regulates the alternative NFkappaB pathway in a ligand-specific manner. PMID- 19809575 TI - The Role of the Theory-of-Mind Cortical Network in the Comprehension of Narratives. AB - Narrative comprehension rests on the ability to understand the intentions and perceptions of various agents in a story who interact with respect to some goal or problem. Reasoning about the state of mind of another person, real or fictional, has been referred to as Theory of Mind processing. While Theory of Mind Processing was first postulated prior to the existence of neuroimaging research, fMRI studies make it possible to characterize this processing in some detail. We propose that narrative comprehension makes use of some of the neural substrate of Theory of Mind reasoning, evoking what is referred to as a protagonist perspective network. The main cortical components of this protagonist based network are the dorsomedial prefrontal cortex and the right temporo parietal junction. The article discusses how these two cortical centers interact in narrative comprehension but still play distinguishable roles, and how the interaction between the two centers is disrupted in individuals with autism. PMID- 19809576 TI - In vitro and In vivo Assessment of CdTe and CdHgTe Toxicity and Clearance. AB - Semiconductor QDs are being developed as fluorescent tags for biomedical applications such as imaging, targeting, therapeutic carriers, drug delivery, nanomedicine, and in vitro and in vivo biological labeling. However, potential toxicity and clearance of semiconductor QDs in biological systems are of concerned. We have tested toxicity and clearance in vitro and in vivo (subcutaneous)of CdTe and CdHgTe semiconductor QDs in human breast cancer MCF7 and MBA-MD-231 cells and prostate cancer PC3 cells over a period of 40 days. Our results show that both CdTe and CdHgTe QDs are cytotoxic to human breast and prostate cancer cells. CdHgTe ODs were cleared rapidly from the site of injection, while CdTe were still detectable 18 days after injection, but were cleared by 30 days. PMID- 19809577 TI - Modulation of the BRCA1 Protein and Induction of Apoptosis in Triple Negative Breast Cancer Cell Lines by the Polyphenolic Compound Curcumin. AB - In the current study, we sought to examine the effects of curcumin in a specific type of breast cancer called triple negative breast cancer. These cancers lack expression of the estrogen and progesterone receptors and do not over-express HER2. Current treatment for triple negative breast cancers is limited to cytotoxic chemotherapy, and upon relapse, there are not any therapies currently available. We demonstrate here that the bioactive food compound curcumin induces DNA damage in triple negative breast cancer cells in association with phosphorylation, increased expression, and cytoplasmic retention of the BRCA1 protein. In addition, curcumin promotes apoptosis and prevents anchorage independent growth and migration of triple negative breast cancer cells. Apoptosis and BRCA1 modulation were not observed in non-transformed mammary epithelial cells, suggesting curcumin may have limited non-specific toxicity. This study suggests that curcumin and potentially curcumin analogues should be tested further in the context of triple negative breast cancer. These results are novel, having never been previously reported, and suggest that curcumin could provide a novel, non-toxic therapy, which could lead to improved survival for patients with triple negative breast cancer. Curcumin should be studied further in this subset of breast cancer patients, for whom treatment options are severely limited. PMID- 19809578 TI - Juvenile Correctional Workers' Perceptions of Suicide Risk Factors and Mental Health Issues of Incarcerated Juveniles. AB - Correctional staff knowledge, attitudes, and perceptions of incarcerated juveniles' mental health needs, including suicide prevention, have not been studied empirically. This study measured juvenile correctional officers' knowledge and attitudes regarding suicide risk factors and mental health and substance abuse issues through administration of the Mental Health Knowledge and Attitude Test (MHKAT) before and after a staff training on suicide prevention. Seventy-six participants completed the pre- and post-training MHKAT. They demonstrated significant improvement in knowledge of and attitudes toward mental health treatment of incarcerated youth as reflected by higher post-training MHKAT scores. Findings suggest that correctional staff are receptive to increasing knowledge of critical mental health issues. Studies of the retention and implementation of this new knowledge by direct care staff over time and the optimal type and frequency of new staff training and continuing education are indicated. PMID- 19809579 TI - The case for chronic disease management for addiction. AB - Chronic disease (care) management (CDM) is a patient-centered model of care that involves longitudinal care delivery; integrated, and coordinated primary medical and specialty care; patient and clinician education; explicit evidence-based care plans; and expert care availability. The model, incorporating mental health and specialty addiction care, holds promise for improving care for patients with substance dependence who often receive no care or fragmented ineffective care. We describe a CDM model for substance dependence and discuss a conceptual framework, the extensive current evidence for component elements, and a promising strategy to reorganize primary and specialty health care to facilitate access for people with substance dependence. The CDM model goes beyond integrated case management by a professional, colocation of services, and integrated medical and addiction care-elements that individually can improve outcomes. Supporting evidence is presented that: 1) substance dependence is a chronic disease requiring longitudinal care, although most patients with addictions receive no treatment (eg, detoxification only) or short-term interventions, and 2) for other chronic diseases requiring longitudinal care (eg, diabetes, congestive heart failure), CDM has been proven effective. PMID- 19809580 TI - Randomized Clinical Trial of Motivational Enhancement of Substance Use Treatment Among Incarcerated Adolescents: Post-Release Condom Non-Use. AB - Evaluated impact of motivational enhancement (ME) of substance abuse treatment compared to relaxation training (RT) on sex without condoms (overall and involving substance use) 3 months following release among incarcerated adolescents. This randomized clinical trial involved 114 incarcerated adolescents from the Northeast. Regression analyses determined if treatment condition, baseline levels of depressive symptoms, and their interaction predicted condom non-use 3 months post-release, controlling for baseline condom non-use. Among those who reported fewer baseline depressive symptoms, those in ME condition reported significantly less condom non-use, in general and involving marijuana use compared with those in RT condition. Periods of incarceration represent opportunities to help juvenile detainees reduce behaviors that impact their health and the health of those with whom they interact in the community. PMID- 19809581 TI - Quality Meshing of Implicit Solvation Models of Biomolecular Structures. AB - This paper describes a comprehensive approach to construct quality meshes for implicit solvation models of biomolecular structures starting from atomic resolution data in the Protein Data Bank (PDB). First, a smooth volumetric electron density map is constructed from atomic data using weighted Gaussian isotropic kernel functions and a two-level clustering technique. This enables the selection of a smooth implicit solvation surface approximation to the Lee Richards molecular surface. Next, a modified dual contouring method is used to extract triangular meshes for the surface, and tetrahedral meshes for the volume inside or outside the molecule within a bounding sphere/box of influence. Finally, geometric flow techniques are used to improve the surface and volume mesh quality. Several examples are presented, including generated meshes for biomolecules that have been successfully used in finite element simulations involving solvation energetics and binding rate constants. PMID- 19809582 TI - Sex-specific costs of reproduction in Eastern Bluebirds Sialia sialis. AB - In species with bi-parental care, individuals must partition energy between parental effort and mating effort. Typically, female songbirds invest more than males in reproductive activities such as egg-laying and incubation, but males invest more in secondary sexual traits used in attracting mates. Animals that breed more than once within a season must also allocate time and energy between first and subsequent breeding attempts and between current and future breeding seasons. To investigate strategies of reproductive investment by males and females and the consequences of such strategies, we manipulated the size of broods of Eastern Bluebirds Sialia sialis. Pairs with enlarged first broods were less likely to produce a second clutch or took longer to initiate one than pairs with reduced broods. After rearing enlarged broods, females were less likely than males to survive to the following year. Although plumage coloration is a sexually selected trait in Eastern Bluebirds that is influenced by nutritional stress, we did not detect an effect of brood-size manipulation on female coloration. Past research, however, demonstrates that, in males, plumage colour is negatively affected by increasing brood size. We suggest that there are sex-specific strategies of reproductive investment in Eastern Bluebirds, and that researchers should incorporate measures of residual reproductive value in studies of life history evolution. PMID- 19809583 TI - Transport of Relational Structures in Groups of Diffeomorphisms. AB - This paper focuses on the issue of translating the relative variation of one shape with respect to another in a template centered representation. The context is the theory of Diffeomorphic Pattern Matching which provides a representation of the space of shapes of objects, including images and point sets, as an infinite dimensional Riemannian manifold which is acted upon by groups of diffeomorphisms. We discuss two main options for achieving our goal; the first one is the parallel translation, based on the Riemannian metric; the second one, based on the group action, is the coadjoint transport. These methods are illustrated with 3D experiments. PMID- 19809584 TI - Measures of emotion: A review. AB - A consensual, componential model of emotions conceptualises them as experiential, physiological, and behavioural responses to personally meaningful stimuli. The present review examines this model in terms of whether different types of emotion evocative stimuli are associated with discrete and invariant patterns of responding in each response system, how such responses are structured, and if such responses converge across different response systems. Across response systems, the bulk of the available evidence favours the idea that measures of emotional responding reflect dimensions rather than discrete states. In addition, experiential, physiological, and behavioural response systems are associated with unique sources of variance, which in turn limits the magnitude of convergence across measures. Accordingly, the authors suggest that there is no "gold standard" measure of emotional responding. Rather, experiential, physiological, and behavioural measures are all relevant to understanding emotion and cannot be assumed to be interchangeable. PMID- 19809585 TI - Role of the PROS1 gene in thrombosis: lessons and controversies. PMID- 19809586 TI - Self-assembly approaches to nanomaterial encapsulation in viral protein cages. AB - A perspective on abiotic material encapsulation inside virus capsids is provided. The emphasis is on the physical principles of virus assembly relevant to packaging, strategies for encapsulation and capsid modification, and on emerging applications. PMID- 19809587 TI - Studies on the Bisoxazoline and (-)-Sparteine Mediated Enantioselective Addition of Organolithium Reagents to Imines. AB - The enantioselective addition of organolithium reagents to N-anisyl aldimines promoted by chiral bisoxazolines and (-)-sparteine as external ligands is described. This reaction proceeds readily with a wide range of aldimine substrates (aliphatic, aromatic, olefinic) and organolithium nucleophiles (Me, n Bu, Ph, vinyl) in excellent yields (81-99%) and with high enantioselectivities (up to 95.5:4.5 er). The external ligands can be used in substoichiometric amounts albeit with slightly attenuated enantioselectivities. A systematic evaluation of the structural features of the bisoxazolines revealed a primary contribution from the substituent at C(4) and a secondary influence from the bridging substituents. A computational analysis (PM3) provided a clear rationalization for the origin of enantioselectivity. PMID- 19809588 TI - Protein Mechanics: A New Frontier in Biomechanics. AB - Proteins play essential roles in all aspects of cellular processes, such as biosynthesis, division, growth, motility, metabolism, signaling, and transmission of genetic information. Proteins, however, could deform under mechanical forces, thus altering their biological functions. Here we present protein deformation as a possible molecular basis for mechanosensing and mechanotransduction, elucidate the important features of protein mechanics including protein deformation mode and dynamics, illustrate how protein deformation could alter biological function, and describe the important roles of protein deformation in force-sensing, force transducing and mechanochemical coupling in cells. The experimental and modeling challenges in protein mechanics are discussed. PMID- 19809589 TI - High-Frequency Oscillations of a Sphere in a Viscous Fluid near a Rigid Plane. AB - High-frequency oscillations of a rigid sphere in an incompressible viscous fluid moving normal to a rigid plane are considered when the ratio of minimum clearance to sphere radius is small. Asymptotic expansions are constructed that permit an analytical estimate of the force acting on the sphere as a result of its motion. An inner expansion, valid in the neighborhood of the minimum gap, reflects the dominance of viscous effects and fluid inertia. An outer expansion, valid outside the gap, reflects the dominance of fluid inertia with a correction for an oscillating viscous boundary layer. The results are applied to the hydrodynamics of the tapping mode of an atomic force microscope and to the dynamic calibration of its cantilevers. PMID- 19809590 TI - Social information guides infants' selection of foods. AB - Two experiments investigated the influence of socially conveyed emotions and speech on infants' preferences in the food domain. After watching films in which two unfamiliar actresses each spoke while eating a different kind of food, 12 month-old infants were allowed to choose between the two foods. In Experiment 1, infants selected a food endorsed by a speaker of their native language who displayed positive affect over a food endorsed by a foreign-language speaker who displayed negative affect. In Experiment 2, both actresses displayed positive affect yet spoke in different languages, and infants again selected the food associated with the speaker of their native language. The findings contrast with previous research in which infants and toddlers have shown little selectivity when presented with foods that differ in their intrinsic properties such as color, texture, and familiarity. Although infants may lack capacities for evaluating foods on their own, they do look to other people for guidance in food selection. PMID- 19809591 TI - Collaborative Behavioral Management for Drug-Involved Parolees: Rationale and Design of the Step'n Out Study. AB - This article describes the rationale, study design, and implementation for the Step'n Out study of the Criminal Justice Drug Abuse Treatment Studies. Step'n Out tests the relative effectiveness of collaborative behavioral management of drug involved parolees. Collaborative behavioral management integrates the roles of parole officers and treatment counselors to provide role induction counseling, contract for pro-social behavior, and deliver contingent reinforcement of behaviors consistent with treatment objectives. The Step'n Out study will randomize 450 drug-involved parolees to collaborative behavioral management or usual parole. Follow-up at 3-and 9-months will assess primary outcomes of rearrest, crime and drug use. If collaborative behavioral management is effective, its wider adoption could improve the outcomes of community reentry of drug-involved ex-offenders. PMID- 19809592 TI - Iron Chelators as Potential Therapeutic Agents for Parkinson's Disease. AB - Parkinson's disease (PD) is a neurological disorder characterized by the progressive impairment of motor skills in patients. Growing evidence suggests that abnormal redox-active metal accumulation, caused by dysregulation, plays a central role in the neuropathology of PD. Redox-active metals (e.g. Fe and Cu) catalyze essential reactions for brain function. However, these metals can also participate in the generation of highly toxic free radicals that can cause oxidative damage to cells and ultimately lead to the death of dopamine-containing neurons. The emergence of redox-active metals as key players in the pathogenesis of PD strongly suggests that metal-chelators could be beneficial in the treatment of this condition. This mini-review summarizes major recent developments on natural, synthetic iron chelating compounds and hydrogen peroxide-triggered prochelators as potential candidates for PD treatment. PMID- 19809594 TI - Lost Mold-Rapid Infiltration Forming of Mesoscale Ceramics: Part 2, Geometry and Strength Improvements. AB - Iterative process improvements have been used to eliminate strength-limiting geometric flaws in mesoscale bend bars composed of yttria-tetragonal zirconia polycrystals (Y-TZP). These improvements led to large quantities of high bend strength material. The metrology of Y-TZP mesoscale bend bars produced using a novel lost mold-rapid infiltration-forming process (LM-RIF) is characterized over several process improvements. These improvements eliminate trapezoidal cross sections in the parts, reduce concave upper surfaces in cross section, and minimize warping along the long axis of 332 x 26 x 17 mum mesoscale bend bars. The trapezoidal cross sections of earlier, first-generation parts were due to the absorption of high-energy ultraviolet (UV) light during the photolithographic mold-forming process, which produced nonvertical mold walls that the parts mirrored. The concave upper surfaces in cross section were eliminated by implementing a RIF-buffing process. Warping during sintering was attributed to impurities in the substrate, which creates localized grain growth and warping as the tetragonal phase becomes destabilized. Precision in the part dimensions is demonstrated using optical profilometry on bend bars and a triangular test component. The bend bar dimensions have a 95% confidence interval of < +/-1 mum, and the tip radius of the triangular test component is 3 mum, consistent with the UV-photolithographic process used to form the mold cavities. The average bend strength of the mesoscale Y-TZP bend exceeds 2 GPa with a Weibull modulus equal to 6.3. PMID- 19809593 TI - PET and SPECT Imaging of Tumor Biology: New Approaches towards Oncology Drug Discovery and Development. AB - Spiraling drug developmental costs and lengthy time-to-market introduction are two critical challenges facing the pharmaceutical industry. The clinical trials success rate for oncology drugs is reported to be 5% as compared to other therapeutic categories (11%) with most failures often encountered late in the clinical development process. PET and SPECT nuclear imaging technologies could play an important role in facilitating the drug development process improving the speed, efficiency and cost of drug development. This review will focus on recent studies of PET and SPECT radioligands in oncology and their application in the investigation of tumor biology. The use of clinically-validated radioligands as imaging-based biomarkers in oncology could significantly impact new cancer therapeutic development. PMID- 19809595 TI - Lost Mold Rapid Infiltration Forming of Mesoscale Ceramics: Part 1, Fabrication. AB - Free-standing mesoscale (340 mum x 30 mum x 20 mum) bend bars with an aspect ratio over 15:1 and an edge resolution as fine as a single grain diameter ( approximately 400 nm) have been fabricated in large numbers on refractory ceramic substrates by combining a novel powder processing approach with photoresist molds and an innovative lost-mold thermal process. The colloid and interfacial chemistry of the nanoscale zirconia particulates has been modeled and used to prepare highly concentrated suspensions. Engineering solutions to challenges in mold fabrication and casting have yielded free-standing, crack-free parts. Molds are fabricated using high-aspect-ratio photoresist on ceramic substrates. Green parts are formed using a rapid infiltration method that exploits the shear thinning behavior of the highly concentrated ceramic suspension in combination with gelcasting. The mold is thermally decomposed and the parts are sintered in place on the ceramic substrate. Chemically aided attrition milling disperses and concentrates the as-received 3Y-TZP powder to produce a dense, fine-grained sintered microstructure. Initial three-point bend strength data are comparable to that of conventional zirconia; however, geometric irregularities (e.g., trapezoidal cross sections) are present in this first generation and are discussed with respect to the distribution of bend strength. PMID- 19809596 TI - Compact integration factor methods in high spatial dimensions. AB - The dominant cost for integration factor (IF) or exponential time differencing (ETD) methods is the repeated vector-matrix multiplications involving exponentials of discretization matrices of differential operators. Although the discretization matrices usually are sparse, their exponentials are not, unless the discretization matrices are diagonal. For example, a two-dimensional system of N * N spatial points, the exponential matrix is of a size of N(2) * N(2) based on direct representations. The vector-matrix multiplication is of O(N(4)), and the storage of such matrix is usually prohibitive even for a moderate size N. In this paper, we introduce a compact representation of the discretized differential operators for the IF and ETD methods in both two- and three-dimensions. In this approach, the storage and CPU cost are significantly reduced for both IF and ETD methods such that the use of this type of methods becomes possible and attractive for two- or three-dimensional systems. For the case of two-dimensional systems, the required storage and CPU cost are reduced to O(N(2)) and O(N(3)), respectively. The improvement on three-dimensional systems is even more significant. We analyze and apply this technique to a class of semi-implicit integration factor method recently developed for stiff reaction-diffusion equations. Direct simulations on test equations along with applications to a morphogen system in two-dimensions and an intra-cellular signaling system in three-dimensions demonstrate an excellent efficiency of the new approach. PMID- 19809597 TI - Molecular simulation of protein-surface interactions: benefits, problems, solutions, and future directions. AB - While the importance of protein adsorption to materials surfaces is widely recognized, little is understood at this time regarding how to design surfaces to control protein adsorption behavior. All-atom empirical force field molecular simulation methods have enormous potential to address this problem by providing an approach to directly investigate the adsorption behavior of peptides and proteins at the atomic level. As with any type of technology, however, these methods must be appropriately developed and applied if they are to provide realistic and useful results. Three issues that are particularly important for the accurate simulation of protein adsorption behavior are the selection of a valid force field to represent the atomic-level interactions involved, the accurate representation of solvation effects, and system sampling. In this article, each of these areas is addressed and future directions for continued development are presented. PMID- 19809598 TI - Molecular packing and magnetic properties of lithium naphthalocyanine crystals: hollow channels enabling permeability and paramagnetic sensitivity to molecular oxygen. AB - The synthesis, structural framework, magnetic and oxygen-sensing properties of a lithium naphthalocyanine (LiNc) radical probe are presented. LiNc was synthesized in the form of a microcrystalline powder using a chemical method and characterized by electron paramagnetic resonance (EPR) spectroscopy, magnetic susceptibility, powder X-ray diffraction analysis, and mass spectrometry. X-Ray powder diffraction studies revealed a structural framework that possesses long, hollow channels running parallel to the packing direction. The channels measured approximately 5.0 * 5.4 A(2) in the two-dimensional plane perpendicular to the length of the channel, enabling diffusion of oxygen molecules (2.9 * 3.9 A(2)) through the channel. The powdered LiNc exhibited a single, sharp EPR line under anoxic conditions, with a peak-to-peak linewidth of 630 mG at room temperature. The linewidth was sensitive to surrounding molecular oxygen, showing a linear increase in pO(2) with an oxygen sensitivity of 31.2 mG per mmHg. The LiNc microcrystals can be further prepared as nano-sized crystals without the loss of its high oxygen-sensing properties. The thermal variation of the magnetic properties of LiNc, such as the EPR linewidth, EPR intensity and magnetic susceptibility revealed the existence of two different temperature regimes of magnetic coupling and hence differing columnar packing, both being one dimensional antiferromagnetic chains but with differing magnitudes of exchange coupling constants. At a temperature of ~50 K, LiNc crystals undergo a reversible phase transition. The high degree of oxygen-sensitivity of micro- and nano-sized crystals of LiNc, combined with excellent stability, should enable precise and accurate measurements of oxygen concentration in biological systems using EPR spectroscopy. PMID- 19809599 TI - Professional Development in Early Childhood Programs: Process Issues and Research Needs. AB - In light of the current policy context, early childhood educators are being asked to have a complex understanding of child development and early education issues and provide rich, meaningful educational experiences for all children and families in their care. Accountability for outcomes is high, and resources for professional support are limited. As such, the early education field needs well conducted empirical studies on which to base professional development practices. In this paper, we offer research directions associated with the processes underlying professional development, including areas in need of investigation that can inform the early childhood education field in terms of how professional development efforts exert their influence and produce meaningful change in practitioners' skills, behaviors, and dispositions. The paper highlights representative research from the professional development literature on its various forms/approaches and offers an agenda for research on the professional development process. Broad issues associated with the conduct of research on professional development, including considerations of professional development processes, participant characteristics, relationships, and sustainability are discussed. PMID- 19809600 TI - Singular perturbation analysis of the steady-state Poisson-Nernst-Planck system: Applications to ion channels. AB - Ion channels are proteins with a narrow hole down their middle that control a wide range of biological function by controlling the flow of spherical ions from one macroscopic region to another. Ion channels do not change their conformation on the biological time scale once they are open, so they can be described by a combination of Poisson and drift-diffusion (Nernst-Planck) equations called PNP in biophysics. We use singular perturbation techniques to analyse the steady state PNP system for a channel with a general geometry and a piecewise constant permanent charge profile. We construct an outer solution for the case of a constant permanent charge density in three dimensions that is also a valid solution of the one-dimensional system. The asymptotical current-voltage (I-V ) characteristic curve of the device (obtained by the singular perturbation analysis) is shown to be a very good approximation of the numerical I-V curve (obtained by solving the system numerically). The physical constraint of non negative concentrations implies a unique solution, i.e., for each given applied potential there corresponds a unique electric current (relaxing this constraint yields non-physical multiple solutions for sufficiently large voltages). PMID- 19809602 TI - Making ethical decisions in current orthopedic practice. PMID- 19809603 TI - Computer-assisted spinal navigation using a percutaneous dynamic reference frame for posterior fusions of the lumbar spine. AB - We report a 6-year retrospective review of screw placement utilizing a percutaneous dynamic reference frame attached to the posterior superior iliac spine performed by a single orthopedic surgeon. We included all lumbar spine procedures utilizing computer-assisted spinal navigation (StealthStation Navigation System, Medtronic Navigation, Louisville, Colo) performed from 2000 to 2005, with 272 of 289 patients (94.1%) having at least a 4-month follow-up with radiographs. Six hundred seventy-two screws were placed. Following surgery, none of these patients had screw misplacements. One patient (0.4%) had a screw backing out of the pedicle. Eighteen patients (6.6%) had their posterior instrumentation removed. Three had repeat operations but did not have their instrumentation removed. No patient with repeat operations had a screw misplaced. PMID- 19809604 TI - Thromboprophylaxis in orthopedic surgery: how long is long enough? AB - Pharmacologic thromboprophylaxis with low-molecular-weight heparins, vitamin K antagonists, or fondaparinux is well tolerated and effective in preventing venous thromboembolism (VTE) in major orthopedic surgery but is often limited to in hospital use. However, 45% to 80% of all symptomatic VTE events occur after hospital discharge. Extended-duration VTE prophylaxis for 28 to 35 days reduces risk for late VTE by up to 70%. In this article, I review the evidence supporting guideline recommendations regarding extended-duration prophylaxis after major orthopedic surgery and provide an overview of current and emerging literature regarding prevention of postoperative VTE in patients undergoing this surgery. PMID- 19809605 TI - Risk of progression in de novo low-magnitude degenerative lumbar curves: natural history and literature review. AB - Natural history studies have focused on risk for progression in lumbar curves of more than 30 degrees, while smaller curves have little data for guiding treatment. We studied curve progression in de novo degenerative scoliotic curves of no more than 30 degrees. Radiographs of 24 patients (17 women, 7 men; mean age, 68.2 years) followed for up to 14.3 years (mean, 4.85 years) were reviewed. Risk factors studied for curve progression included lumbar lordosis, lateral listhesis of more than 5 mm, sex, age, convexity direction, and position of intercrestal line. Curves averaged 14 degrees at presentation and 22 degrees at latest follow-up and progressed a mean of 2 degrees (SD, 1 degrees) per year. Mean progression was 2.5 degrees per year for patients older than 69 years and 1.5 degrees per year for younger patients. Levoscoliosis progressed 3 degrees per year and dextroscoliosis 1 degrees per year (P<.05). Forty-six percent of patients had lateral listhesis of more than 5 mm at L3 and L4. Curve progression was not linear and might occur rapidly, particularly in women older than 69 with lateral listhesis of more than 5 mm and levoscoliosis. Small curves can progress and therefore should be individualized in the context of other risk factors. PMID- 19809606 TI - Treatment of displaced type II odontoid fractures in elderly patients. AB - Odontoid fractures are the most common cervical spine fractures for patients older than 70 years and are the most common of all spinal fractures for patients older than 80. Type II fracture, the most common type of odontoid fracture, is considered relatively unstable. It occurs at the base of the odontoid between the level of the transverse ligament and the C2 vertebral body. In the geriatric population, it is important to look for any associated clinical comorbidities that might affect management. Treatment options for displaced odontoid fractures can be conservative or surgical. Conservative management includes immobilization in a cervical collar or in a halo vest. External immobilization with a cervical collar has had inconsistent results. Halo vest immobilization in the elderly is associated with a significant nonunion rate and several complications. Generally accepted surgical indications are polytrauma, neurologic deficit, associated unstable subaxial spine injury that requires surgical fixation, and symptomatic nonunion. Surgical management includes either anterior odontoid screw fixation or posterior C1-C2 instrumentation with fusion. PMID- 19809607 TI - Word of mouth in the digital age: online physician ratings. PMID- 19809608 TI - Analgesia for total hip and knee arthroplasty: a review of lumbar plexus, femoral, and sciatic nerve blocks. AB - Use of peripheral nerve blocks (PNBs) during lower extremity surgery has evolved. In this article, we review the pertinent anatomy and the literature concerning the advantages and disadvantages of both PNBs and traditional methods of postoperative analgesia (neuraxial and patient-controlled) for total hip arthroplasty and total knee arthroplasty. We conclude that use of PNBs for total hip and total knee arthroplasty compares favorably with traditional methods of postoperative analgesia. As use of PNBs becomes more widespread, understanding their risks and benefits will be of great value to orthopedic surgeons. PMID- 19809609 TI - Blood cultures for evaluation of fever after total joint arthroplasty. AB - Fever after total joint arthroplasty (TJA) is common. Fearing the potential complications of bacteremia, physicians often obtain blood cultures to evaluate fever after TJA. In this study, we retrospectively examined the results of 102 sets (204 samples) of blood cultures that had been obtained from 50 patients (mean age, 67.3 years) during the first 2 postoperative days for evaluation of fever of 38.3 degrees C or higher. All patients had been receiving antibiotic prophylaxis. Of the 50 patients, 39 had undergone total knee arthroplasty, and 11 had undergone total hip arthroplasty. There had been 49 primary operations and 1 revision. Of the 204 blood culture samples, none had grown a pathogen. The cultures had been ordered by both surgical (61%) and medical (39%) services. The 2008 institution charge to process the 2 blood culture samples (1 set) routinely collected for each evaluation was $120 (true cost, $44.29). Therefore, in the current healthcare market, the charge to the payer for processing 102 sets would be $12,240. We conclude that blood cultures are neither useful nor cost-effective in evaluating fever immediately after TJA. We believe that the results of this study will be helpful to both orthopedists and medical consultants involved in the care of TJA patients. PMID- 19809610 TI - Venous hemodynamic alterations in lower limbs undergoing total joint arthroplasty. AB - Using duplex ultrasonography, we measured preoperative and postoperative venous flow volume in 32 operated lower limbs without deep vein thrombosis (DVT) after total hip arthroplasty (THA, n = 17) and total knee arthroplasty (TKA, n = 15). We also calculated percentage decrease in mean venous flow volume (MVFV) from before surgery to after surgery. Patients with a history of one of several venous diseases, congestive heart failure, or morbid obesity were excluded. In both groups (THA, TKA), MVFV 3 days after surgery and MVFV 1 week after surgery were significantly lower than preoperative MVFV, but MVFV at 2 or more weeks after surgery did not differ significantly from preoperative MVFV (result 1). Incidentally, the decrease in MVFV in the lower limbs was significantly larger 3 days after TKA than 3 days after THA (result 2). As venous stasis has a central role in thrombus formation, result 1 suggests that the risk for DVT initiation is low at 2 or more weeks after THA and TKA in patients with normal preoperative venous physiologic functions. Result 2 is probably correlated with the evidence that DVT incidence is higher after TKA than after THA. PMID- 19809611 TI - Joint arthroplasty within 10 years after primary charnley total hip arthroplasty. AB - To evaluate the need for joint arthroplasty within 10 years after index primary Charnley total hip arthroplasty (THA) performed for osteoarthritis, we retrospectively reviewed the cases of 2,547 patients, 50 to 75 years old, from 1969 to 1984, with a minimum potential 20-year follow-up. In this article, we report the age, sex, and time data from this study. For the entire patient population, the 10-year rate of undergoing contralateral THA was 35.0%; ipsilateral hip revision, 6.2%; ipsilateral total knee arthroplasty (TKA), 0.6%; contralateral TKA, 1.9%; and bilateral TKA, 0.2%. The 10-year death rate was 21.8%. With more than 200,000 THAs being performed in the United States each year, these numbers can guide orthopedic surgeons in their discussions about subsequent arthroplasty procedures on other joints. PMID- 19809615 TI - Controlling phase and morphology of inorganic nanostructures originated from the internal crystal structure. AB - Synthesis of inorganic nanostructures designed from the internal crystal structure has been a subject of intensive research in the past decade. By virtue of structural understanding, attention was devoted to novel strategies to control morphology and phase formation of nanostructures, and even their assembly to hierarchitectures. Also, the structural analysis gives an alternative way to understand or predict the potential physicochemical properties of inorganic solids. In this article our recent advances in nanostructure generation and their physicochemical-property understanding from crystal structures are highlighted. PMID- 19809616 TI - Probing molecular chirality by CD-sensitive dimeric metalloporphyrin hosts. AB - This feature article reviews dimeric metalloporphyrin hosts employed as chirality probes in chiral recognition processes involving synthetic compounds and natural products. Upon formation of a chiral host-guest supramolecular complex between an achiral bis-metalloporphyrin derivative and a chiral non-racemic guest, a CD response occurs in the porphyrin spectral region, which is diagnostic of the guest's absolute configuration. Several bis-porphyrin hosts used in the stereochemical investigation of organic compounds are described and the scope of their application as chirality probes critically assessed. The review encompasses the description of structural features of the host-guest complexes, the nature of the chirality transfer mechanism and the practical application in solving stereochemical problems. In particular, with reference to the method based on bis porphyrin tweezers, we describe recent advances based on the use of molecular modeling, which have broadened the applicability of the tweezer methodology and allowed extraction of deeper structural information contained in the experimental CD data. PMID- 19809617 TI - Towards a temperature-guided molecular switch: an unusual reversible low temperature polymorphic phase transition in a conformationally locked environment. AB - A conformationally locked tetraacetate undergoes, quite akin to a temperature guided molecular switch, a reversible thermal switching between two polymorphic modifications; the room-temperature alpha-form converted at -4 degrees C to a low temperature denser beta-form, which displayed an unusual kinetic stability till 67 degrees C and transformed back to the alpha-form beyond this temperature. PMID- 19809618 TI - Role of complementary H-bonding interaction of a cyanurate in the self-assembly and gelation of melamine linked tri(p-phenyleneethynylene)s. AB - Melamine-functionalized tri(p-phenyleneethynylene) self-assembles to form opaque and weak gels in aliphatic solvents which turned transparent and stable upon addition of a cyanurate, affording supramolecular nanostructures with distinct physical properties. PMID- 19809619 TI - Asymmetric beta-boration of alpha,beta-unsaturated carbonyl compounds promoted by chiral rhodium-bisoxazolinylphenyl catalysts. AB - Chiral rhodium-bisoxazolinylphenyl acetate complexes exhibited high catalytic activity for the beta-boration of alpha,beta-unsaturated carbonyl compounds with bis(pinacolato)diboron in the presence of sodium t-butoxide with enantioselectivity up to 97%. PMID- 19809620 TI - Magnetic nanoparticle-supported Hoveyda-Grubbs catalysts for ring-closing metathesis reactions. AB - Magnetically recyclable Hoveyda-Grubbs catalyst can be readily assembled using magnetic nanoparticles as support, and this catalyst combines convenient recyclability and excellent activity on ring-closing metathesis (RCM) reactions. PMID- 19809621 TI - Acid-mediated formation of trifluoromethyl sulfonates from sulfonic acids and a hypervalent iodine trifluoromethylating agent. AB - A variety of sulfonic acids have been trifluoromethylated using 1-trifluoromethyl 1,2-benziodoxol-3(1H)-one under mild conditions in good to excellent yields. Initial mechanistic investigations of this reaction show a clean second-order kinetics and only very weak substrate electronic effects. PMID- 19809622 TI - Iridium-catalyzed enantioselective hydrogenation of vinyl boronates. AB - The first Ir-catalyzed asymmetric hydrogenations of vinyl boronates have been performed using low catalyst loadings (0.5 mol%) and pressure (as low as 1 bar). Good selectivities (76-98% ee) were obtained for a range of substrates. PMID- 19809623 TI - Ultra-short contact time conversion of chloromethane to olefins over pre-coked SAPO-34: direct insight into the primary conversion with coke deposition. AB - Hydrocarbon species trapped in the catalyst that work as the reaction center and govern the conversion and product selectivity of chloromethane transformation were revealed by a very efficient home-made ultra-short contact time pulse reaction system over pre-coked SAPO-34. PMID- 19809624 TI - Direct functionalization of benzylic C-Hs with vinyl acetates via Fe-catalysis. AB - Direct cross-coupling to construct sp3 C-sp3 C bonds via Fe-catalyzed benzylic C H activation with 1-aryl vinyl acetate was developed. PMID- 19809625 TI - Supramolecular aggregation of functionalized carbon nanotubes. AB - We describe the synthesis of novel carbon nanotubes functionalized with thymine units, able to induce self-assembly in a controlled supramolecular fashion. PMID- 19809626 TI - Fabrication of TaS2 nanobelt arrays and their enhanced field-emission. AB - Crystal TaS2 nanobelt arrays were fabricated on Ta wafers by a two-step method. Field-emission measurements show that the nanobelt arrays are decent field emitters with a turn-on field of approximately 0.7 V microm(-1), a threshold field of approximately 2.1 V microm(-1), and a field enhancement factor of approximately 4.33 x 10(4). PMID- 19809627 TI - Non-catalytic conversion of C-F bonds of benzotrifluorides to C-C bonds using organoaluminium reagents. AB - A facile method for the conversion of C-F bonds of benzotrifluorides to C-C bonds has been developed using aluminium reagents in the absence of catalysts. PMID- 19809628 TI - Energetic polyazole polynitrobenzenes and their coordination complexes. AB - New energetic polyazole polynitrobenzenes exhibit high heats of formation and thermal stabilities, and react as useful ligands with silver dinitramide to give an energetic coordination complex. PMID- 19809629 TI - Magnetic peptide nucleic acids for DNA targeting. AB - A versatile synthetic platform for the efficient immobilization of PNAs on magnetic iron oxides, providing magnetic nanosensors for selective DNA recognition, is presented. PMID- 19809630 TI - Controlled preparation of a heterometallic lanthanide complex containing different lanthanides in symmetrical binding pockets. AB - A heterometallic lanthanide complex has been prepared by sequential deprotection and complexation of an orthogonally protected ligand: luminescence and NMR spectroscopy have been used to probe the integrity of the complex. PMID- 19809631 TI - Iron-catalyzed dehydrogenative phosphonation of N,N-dimethylanilines. AB - Iron(II) and iron(III) salts catalyze the oxidative alpha-phosphonation of N,N dimethylanilines with dialkyl H-phosphonates in the presence of tert butylhydroperoxide. PMID- 19809632 TI - Zr-promoted linear coupling of alkynes to generate bis(allene)s. AB - Zirconium promoted linear coupling of alkynes to generate bis(allene)s with high yields in the presence of nBuLi and chlorophosphines or trimethylsilyl trifluoromethanesulfonate. PMID- 19809633 TI - One-pot synthesis and biofunctionalization of glycopolymers via RAFT polymerization and thiol-ene reactions. AB - A modification of a polymeric activated ester with sugar-amines and a simultaneous aminolysis-thiol-ene reaction involving the RAFT end-groups yield biotin functionalized glycopolymer in one-pot. PMID- 19809634 TI - Visible-light-harvesting periodic mesoporous organosilica. AB - Highly ordered periodic mesoporous organosilica synthesized from a newly designed 9(10H)-acridone bridged organosilane precursor exhibited efficient light harvesting antenna properties for visible light, at wavelengths up to 450 nm. PMID- 19809635 TI - Cage connectivities of C88 (33) and C92 (82) fullerenes captured as trifluoromethyl derivatives, C88(CF3)18 and C92(CF3)16. AB - The trifluoromethylation of a higher fullerene mixture followed by HPLC separation allowed the isolation and X-ray structural characterization of C1 C88(CF3)18 and D2-C92(CF3)16 compounds that turned out to be the derivatives of C2-C88 (isomer 33) and D2-C92 (isomer 82), respectively. PMID- 19809636 TI - Multicompartment micelles from a metallo-supramolecular tetrablock quatercopolymer. AB - Multicompartment micelles with tunable compartment solubility have been prepared in alcohols from a metallo-supramolecular poly(p-tetrafluoromethylstyrene)-block poly(tert-butylacrylate)-block-poly(styrene)-[Ru]-poly(ethylene glycol) tetrablock quaterpolymer. PMID- 19809637 TI - Light responsive protective coatings. AB - We demonstrate a strategy of incorporation and opening of light-sensitive containers in anticorrosion coatings for construction of very efficient light responsive protective coatings. PMID- 19809638 TI - Poly-beta-cyclodextrin based platform for pH mapping via a ratiometric 19F/1H MRI method. AB - The in vitro validation of a new pH mapping MRI method based on a supramolecular poly-beta-cyclodextrin-19F-Gd adduct is reported. PMID- 19809639 TI - Multiple conformational changes of beta-tetraphenyl meso hexakis(pentafluorophenyl) substituted [26] and [28]hexaphyrins(1.1.1.1.1.1). AB - Beta-tetraphenyl meso-hexakis(pentafluorophenyl) substituted [26]hexaphyrin is conformationally flexible between rectangular and figure-of-eight shapes and its two-electron reduced [28]hexaphyrin takes figure-of-eight conformations, which are changed, upon protonation, to twisted conformations with distinct Mobius aromaticity. PMID- 19809640 TI - A sensitive and specific electrochemiluminescent sensor for lead based on DNAzyme. AB - A specific ECL sensor for Pb2+ based on DNAzyme has been developed for the first time; the detection limit of 1.1 x 10(-11) mol l(-1) is much lower than those of fluorescent, colorimetric or electrical biosensors. PMID- 19809641 TI - A method for covering a substrate with highly-oriented single crystalline hexagonal zinc structures under ambient pressure and room temperature. AB - We report a novel method for covering a substrate with highly-oriented single crystalline hexagonal zinc structures under atmospheric pressure and room temperature without an external source of electric current, any templates or the use of epitaxial growth on the substrate. PMID- 19809642 TI - Metal-organic framework (MOF) aerogels with high micro- and macroporosity. AB - Iron-BTC aerogels (BTC = 1,3,5-benzene tricarboxylate) with high permanent porosity and total pore volumes of up to 5.6 cm3 g(-1) were obtained in a sol-gel approach followed by supercritical CO2 drying. PMID- 19809643 TI - Stable silver nanoparticle-DNA conjugates for directed self-assembly of core satellite silver-gold nanoclusters. AB - We report a novel strategy to functionalize silver nanoparticles with chimeric phosphorothioate modified DNA (ps-po-DNA) and the fabrication of bimetallic core satellite nanoclusters that each contain a silver core (diameter 32 nm) surrounded by 5 nm gold NPs. PMID- 19809644 TI - Straightforward synthesis of new polyoxometalate-based hybrids exemplified by the covalent bonding of a polypyridyl ligand. AB - A new polyoxometalate-based organic-inorganic platform has been designed for further facile derivatization and covalent attachment of organic linkers; this is exemplified by the grafting of a polypyridyl ligand. PMID- 19809645 TI - Core-shell particle interconversion with di-stimuli-responsive diblock copolymers. AB - Core-shell reversible particle precipitation from aqueous di-stimuli-responsive diblocks is demonstrated as also is the interconversion from one core-shell combination to the other. PMID- 19809646 TI - Facile synthesis of responsive nanoparticles with reversible, tunable and rapid thermal transitions from biocompatible constituents. AB - Responsive polymeric nanoparticles composed of hybrid block co-polymers were prepared from biocompatible components that displayed rapid, tunable and multiply reversible transitions in response to change of temperature. PMID- 19809647 TI - Hollow mesoporous carbon spheres--an excellent bilirubin adsorbent. AB - Hollow mesoporous carbon spheres (HMCSs) were used for the first time as a bilirubin adsorbent, which showed an extraordinarily high bilirubin adsorption capacity as compared with commercial activated carbon for haemoperfusion, high bilirubin adsorption selectivity and negligible hemolytic activity. The results are very attractive and promising for blood purification applications. PMID- 19809648 TI - A novel smart organogel which could allow a two channel anion response by proton controlled reversible sol-gel transition and color changes. AB - A super gelator (F3) with an excellent gelation ability was synthesized; the organogel of F3 could allow two channel recognition of F-, AcO- and H2PO4- through proton controlled reversible sol-gel transition and color changes. PMID- 19809649 TI - The preparation of the MF6- (M = As, Sb) salts of 1,4-benzoquinodal bridged bis 1,3,2-dithiazolylium utilizing the cycloaddition and oxidative dehydrogenation chemistry of SNSMF6 and observation of a hybrid semiquinoidal-thiazyl radical cation by EPR. AB - SNSMF6 (M = As, Sb) reacts with 1,4-benzoquinone in a series of cycloaddition and oxidative dehydrogenation reactions to give 3 (MF6)2 which on reduction with CsI yields the unique semiquinodal-thiazyl radical-cation (3+*) as observed by X-band solution ESR and supported by DFT calculations. PMID- 19809650 TI - Chemical etching with tetrafluoroborate: a facile method for resizing of CdTe nanocrystals under mild conditions. AB - This communication describes a facile and mild chemical etching method for resizing of CdTe nanocrystals, which provides a possible approach for tailoring the size-dependent properties of the nanocrystals. PMID- 19809651 TI - Easy preparation of a reticular nickel film deposited on a barrel-plating gold electrode with a high catalytic activity towards the oxidation of glucose. AB - A reticular and porous Ni film was deposited on a barrel-plating gold electrode by an easy approach and was demonstrated to significantly enhance the catalytic activity towards glucose oxidation. PMID- 19809652 TI - Control of polythiophene redox potentials based on supramolecular complexation with helical schizophyllan. AB - A novel method to control polythiophene redox potentials based on supramolecular complexation with the native polysaccharide, schizophyllan (SPG) is reported, which can importantly improve air stability for easy handling and processing. PMID- 19809653 TI - PPh3-catalyzed unexpected alpha-addition reaction of 1-(o-hydroxyaryl)-1,3 diketones to terminal alkynoates: a straightforward synthesis of multifunctional vinylesters. AB - PPh3-catalyzed alpha-addition reactions of 1-(o-hydroxyaryl)-1,3-diketones to terminal alkynoates involving carbon-carbon bond cleavage to give multifunctional vinylesters are described. PMID- 19809654 TI - Pyrogallarene-based ion-conducting pores that show reversible conductance properties. AB - Pyrogallol[4]arene macrocycles prepared from pyrogallol and n-dodecanal insert in phospholipid bilayers and form conducting pores that undergo reversible switching over a wide range of potentials. PMID- 19809655 TI - Clean and highly ordered graphene synthesized in the gas phase. AB - We report that the substrate-free gas-phase graphene synthesis method produces clean and highly ordered graphene sheets that are similar in quality to the graphene obtained through the mechanical exfoliation of highly oriented pyrolytic graphite. PMID- 19809656 TI - Redox-noninnocence of N,N'-bis(6-methyl-2-pyridylmethylene)ethane-1,2-diamine (L): synthesis and characterization of diamagnetic [Ni(II)2(L**)2] and [Zn(II)2(L)Cl4]((L**)2- pi diradical dianion of L). AB - The synthesis of two diamagnetic dinuclear complexes [Ni(II)2(L**)2] (1) and [Zn(II)2(L)Cl4] (2) and an octahedral, paramagnetic, mononuclear complex [Fe(II)(L)Cl2] (3) (S = 2) is reported; the tetradentate ligand N,N'-bis(6-methyl 2-pyridylmethylene)ethane-1,2-diamine, L, is shown to be redox noninnocent. It exists in its neutral closed-shell form L in 2 and 3 and in its dianionic diradical form, (L**)2-, in 1. PMID- 19809662 TI - Magneto-optical FeGa2O4 nanoparticles as dual-modality high contrast efficacy T2 imaging and cathodoluminescent agents. AB - The magneto-optical FeGa(2)O(4) nanoparticles with high Fe mass magnetization exhibited a high-performance T(2)-lowering effect and displayed cathodoluminescence. PMID- 19809663 TI - Solvent-shift Monte Carlo: a cluster algorithm for solvated systems. AB - We present a cluster algorithm for the efficient simulation of solvated systems that we term solvent-shift Monte Carlo (SSMC). The algorithm involves a conformational change in a solvated solute molecule of interest, followed by a concerted movement of solvent particles about a rotation axis. The method satisfies detailed balance and can be applied to existing schemes to sample conformational space, where an axis or plane of rotation can be defined. We demonstrate that the algorithm significantly enhances the sampling of phase space in solvated systems, and may be easily combined with other advanced sampling techniques. PMID- 19809664 TI - The effect of the NH2 substituent on NH3: hydrazine as an alternative for ammonia in hydrogen release in the presence of boranes and alanes. AB - Potential energy surfaces for H(2) release from hydrazine interacting with borane, alane, diborane, dialane and borane-alane were constructed from MP2/aVTZ geometries and zero point energies with single point energies at the CCSD(T)/aug cc-pVTZ level. With one borane or alane molecule, the energy barrier for H(2) loss of approximately 38 or 30 kcal mol(-1) does not compete with the B-N or Al-N bond cleavage ( approximately 30 or approximately 28 kcal mol(-1)). The second borane or alane molecule can play the role of a bifunctional catalyst. The barrier energy for H(2)-elimination is reduced from 38 to 23 kcal mol(-1), or 30 to 20 kcal mol(-1) in the presence of diborane or dialane, respectively. The mixed borane-alane dimer reduces the barrier energy for H(2) release from hydrazine to approximately 17 kcal mol(-1). A systematic comparison with the reaction pathways from ammonia borane shows that hydrazine could be an alternative for ammonia in producing borane amine derivatives. The results show a significant effect of the NH(2) substituent on the relevant thermodynamics. The B N dative bond energy of 31 kcal mol(-1) in NH(2)NH(2)BH(3) is approximately 5 kcal mol(-1) larger than that of the parent BH(3)NH(3). The higher thermodynamic stability could allow hydrazine-borane to be used as a material for certain energetic H(2) storage applications. PMID- 19809665 TI - Translational, rotational and vibrational energy partitioning in the sequential loss of carbon dimers from fullerenes. AB - The sequential thermal dissociation of up to five carbon dimers from neutral, singly and doubly charged C(60) molecules is theoretically investigated in the framework of phase space theory. Using a semiclassical treatment of vibrations and rotations, we quantify the amount of kinetic energies released in the form of translation, rotation, and vibration under realistic experimental conditions. Our results reveal that translational and vibrational energies of the dimers are nearly equilibrated after a few emissions, whereas the rotational contribution lies far below equipartition. An approximate treatment in which dimers are rotationally and vibrationally frozen essentially leads to the same conclusions. PMID- 19809666 TI - Structure of endohedral fullerene Eu@C74. AB - Structure determination of endohedral fullerenes in the absence of X-ray data is difficult and often controversial. Here we show that the structure of endohedral fullerene Eu@C(74) may be determined by density functional theory aided interpretation of its electronic, infrared and Raman spectra. The use of recently developed analytical polarizability gradient methods to simulate resonance enhanced Raman spectra is crucial for this approach and allows for a nearly complete assignment of the experimental spectra. Eu@C(74) is assigned a pear shaped C(2v) symmetric structure and shows strong ionic interaction between the encapsulated metal and the fullerene pi system. PMID- 19809667 TI - First-principles simulations of hydrogen peroxide formation catalyzed by small neutral gold clusters. AB - Energetics and dynamical pathways for hydrogen peroxide formation from H(2) and O(2) bound to neutral gold dimers and tetramers have been investigated by applying several strategies: T = 0 K geometry optimizations, constrained Car Parrinello molecular dynamics simulations at T = 300 K and metadynamics at T = 300 K. The competing reaction channels for water and hydrogen peroxide formation have been found and characterized. In each case, the reaction barriers for Au cluster catalyzed proton transfer are less than 1 eV. Water formation is a competitive reaction channel, and the relative weight of H(2)O and H(2)O(2) products may depend on the chosen Au cluster size. Dynamic simulations demonstrate the significance of the geometric fluxionality of small catalytic Au clusters. These results indicate that neutral Au clusters could work as catalysts in aerobic H(2)O(2) formation in ambient conditions. PMID- 19809668 TI - Optimal construction of a fast and accurate polarisable water potential based on multipole moments trained by machine learning. AB - To model liquid water correctly and to reproduce its structural, dynamic and thermodynamic properties warrants models that account accurately for electronic polarisation. We have previously demonstrated that polarisation can be represented by fluctuating multipole moments (derived by quantum chemical topology) predicted by multilayer perceptrons (MLPs) in response to the local structure of the cluster. Here we further develop this methodology of modeling polarisation enabling control of the balance between accuracy, in terms of errors in Coulomb energy and computing time. First, the predictive ability and speed of two additional machine learning methods, radial basis function neural networks (RBFNN) and Kriging, are assessed with respect to our previous MLP based polarisable water models, for water dimer, trimer, tetramer, pentamer and hexamer clusters. Compared to MLPs, we find that RBFNNs achieve a 14-26% decrease in median Coulomb energy error, with a factor 2.5-3 slowdown in speed, whilst Kriging achieves a 40-67% decrease in median energy error with a 6.5-8.5 factor slowdown in speed. Then, these compromises between accuracy and speed are improved upon through a simple multi-objective optimisation to identify Pareto optimal combinations. Compared to the Kriging results, combinations are found that are no less accurate (at the 90th energy error percentile), yet are 58% faster for the dimer, and 26% faster for the pentamer. PMID- 19809669 TI - Role of vibrational anharmonicity in atmospheric radical hydrogen-bonded complexes. AB - Harmonic and anharmonic vibrational frequency calculations are reported for the most stable hydrogen bonded complexes formed between the hydroperoxyl radical and formic, acetic, nitric, and sulfuric acids which are of atmospheric interest. A comparison between the calculated IR spectra of the hydrogen bonded complexes with the corresponding separate monomers is also reported with the aim to facilitate a possible experimental identification of these complexes. The calculations have been carried out using the second-order vibrational perturbative treatment implemented by Barone applied to the PES obtained with the B3LYP functional using the 6-31+G(d,p) and 6-311+G(2d,2p) basis sets. Our calculations for the separate monomers predict vibrational frequencies with quite a good agreement with the experimental values. The anharmonic contribution results in differences of around 40 cm(-1) with respect to the harmonic values; although in some cases involving highly anharmonic modes, these differences can rise up to 300 and 450 cm(-1). PMID- 19809670 TI - Vibrational spectroscopy and DFT calculations of the di-amino acid peptide L aspartyl-L-glutamic acid in the zwitterionic state. AB - Solid state IR and Raman as well as aqueous solution state Raman spectra are reported for the linear di-amino acid peptide L-aspartyl-L-glutamic acid (L-Asp-L Glu); the solution state Raman spectrum has also been obtained for the N,O deuterated derivative. SCF-DFT calculations at the B3-LYP/cc-pVDZ level established that the structure and vibrational spectra of L-Asp-L-Glu can be interpreted using a model of the peptide with ten hydrogen-bonded water molecules, in conjunction with the conductor-like polarizable continuum solvation method. The DFT calculations resulted in the computation of a stable zwitterionic structure, which displays trans-amide conformation. The vibrational spectra were computed at the optimised molecular geometry, enabling normal coordinate analysis, which yielded satisfactory agreement with the experimental IR and Raman data. Computed potential energy distributions of the normal modes provided detailed vibrational assignments. PMID- 19809671 TI - Facile method for spectroscopic examination of radical ions of hydrophilic carotenoids. AB - Hydrophilic carotenoids, unusual members of an intrinsically hydrophobic family, and their radical ions are important reactants. An all-optical method for generating singly charged radical ions of a hydrophilic carotenoid (Car) is described. It relies on photolyzing an aqueous mixture of Car and a photoionizable auxiliary solute (A), and making conditions conducive to the capture, by Car, of the hydrated electron (e(aq)(-)) or the positive hole in A(*)(+) or both. When A is Trolox (TOH), only e(aq)(-) can be captured, since TOH (*)(+) deprotonates too rapidly to be a hole donor; when A is Trolox methyl ether (TOMe), both Car(*)(-) and Car(*)(+) are formed, since TOMe (+) lives long enough to transfer its positive hole to Car; formation of Car(*)(-) is prevented under aerobic conditions. PMID- 19809672 TI - Acceleration of the Z to E photoisomerization of penta-2,4-dieniminium by hydrogen out-of-plane motion: theoretical study on a model system of retinal protonated Schiff base. AB - We report the result of comparison between two reaction coordinates [on the potential energy surface of the first excited state (S(1))] produced by CASSCF and these energies recalculated by MRMP2 in the Z to E photoisomerization of penta-2,4-dieniminium (PDI) as the minimal model of the retinal protonated Schiff base (RPSB). One coordinate is the S(1) state minimum-energy-path (MEP) in mass weighted coordinates from the S(1) vertically excited point, where a strong hydrogen-out-of plane (HOOP) motion is not exhibited. The energy profile of the S(1) MEP at the MRMP2//CASSCF level shows a barrier for the rotation around the reactive C-C and hits the S(1)/S(0) degeneracy space where the central C-C-C-C dihedral angle is distorted by 65 degrees . The other coordinate is an S(1) coordinate obtained by the relaxed scan strategy. The relaxed coordinate along the central C-C-C-C dihedral angle, which we call the HOOP coordinate, shows strong HOOP motion. According to the MRMP2//CASSCF calculation, there is no barrier on the HOOP coordinate. Furthermore, the S(1) to S(0) transition may be possible without the large skeletal deformation by HOOP motion because the HOOP coordinate encounters the S(1)/S(0) degeneracy space where the central C-C-C-C dihedral angle is distorted by only 40 degrees . Consequently, if PDI is a suitable model molecule for the RPSB as often assumed, the 11-cis to all-trans photoisomerization is predicted to be accelerated by the HOOP motion. PMID- 19809673 TI - Diode laser based studies of the UV photolysis of molecular iodine. AB - The photolysis of molecular iodine at 193 and 248 nm has been studied by diode laser based frequency-modulated (FM) absorption spectroscopy with detection of the nascent iodine photofragment via the I((2)P(1/2)-(2)P(3/2)) transition at 1.315 microm. Use of narrow band radiation enables nascent measurements with sufficient speed resolution to allow both the character of the initial electronic transition and speed of the fragments to be determined. The time dependence of the integrated area of the measured Doppler profiles has been used to determine both the I* quantum yield and the collisional electronic quenching rate constant of I* (I* = (2)P(1/2)) by I(2). These values are also determined using the diode laser gain versus absorption technique. In the 248 nm case an I* quantum yield of 0.45 +/- 0.04 and 0.42 +/- 0.04 is found by each method, respectively, and an electronic quenching rate constant of (3.6 +/- 0.5) x 10(-11) cm(3) molecule(-1) s(-1), consistent with literature, is determined. The form of the nascent Doppler profile indicates that excitation to a Omega = 1 state dominates, with subsequent dissociation to I((2)P(1/2)) + I((2)P(3/2)), in keeping with assignment of the upper state as 1441 (3)Sigma(+)(1(u)). The deviation from Phi(I*) = 0.5 can be attributed to a contribution from the 1441 (3)Sigma(+)(0(u)(-)) state which dissociates to two ground state iodine atoms. 193 nm excitation exhibits more complicated dynamics and kinetics, including a pressure dependent I* quantum yield. At the low pressures, <200 mTorr, used in the FM Doppler measurements there are two speed components to the profile suggesting multiple dissociation pathways. Firstly, fast iodine fragments indicate single photon absorption in a parallel transition followed by direct dissociation. Secondly, a slower speed component can be attributed to iodine atoms formed after radiative transfer to unbound levels in the ground state and the a' surface. Investigation of the changes in Doppler profiles with time suggest that some of the a' surface population may transfer to the bound B((3)Pi(0u)(+)) state which then in turn undergoes collision-induced predissociation to produce I atoms on the ground electronic state. PMID- 19809674 TI - On the chemical potential of a component in a metastable phase--application to Li storage in the RuO2-Li system. AB - The chemical potential of a component of a binary metastable compound is considered in the single-phase and in the compositionally non-variant two-phase regime. A detailed thermodynamic analysis reveals striking differences for identical nominal compositions. Without the loss of generality the chemical potential of Li in Li containing compounds is referred to. In the single-phase regime an increased Gibbs energy of the metastable phase, compared with the stable phase, leads to an increased chemical potential of lithium, as long as we can ignore the dependence of the excess value on composition. If, in the same approximation, this metastable phase is in two-phase equilibrium with a binary phase more rich in Li, a decrease in the chemical potential and hence an inversion of the sign of the cell voltage in a Li-based battery is predicted. To be specific we consider Li-storage within RuO(2) as well as in the pseudo-binary two-phase system RuO(2)-LiRuO(2). A significantly greater cell voltage vs. Li is observed in the two-phase region if amorphous RuO(2) is used instead of crystalline RuO(2), in contrast to the single-phase system. A possible applicability of metastable phases for Li-based batteries is discussed. PMID- 19809675 TI - Spectroscopic study of electron and energy transfer in novel silicon phthalocyanine--boron dipyrromethene triads. AB - Phthalocyanines (Pcs) and boron dipyrromethenes (BDPs) are two versatile classes of functional dyes suitable for design of artificial light harvesting and charge separation systems. In the present work, we report the results of photophysical investigations of two novel non-sandwich-type BDP-Pc heterotriads, in which two BDP or mono-styryl BDP moieties are linked to the central atom of a silicon(iv) phthalocyanine core (triad 4 and 5, respectively). It was found that the photophysical properties of the triads in toluene and N,N-dimethylformamide (DMF) are strongly affected by two different types of interaction between the BDP and the Pc parts, namely excitation energy transfer (EET) and photoinduced charge transfer (CT). The first process delivers the excitation to the first excited singlet state of the Pc-part upon initial BDP-part excitation. The probability of EET supersedes that of CT in toluene, whereas the latter transfer process dominates in BDP-part depopulation when triads are dissolved in polar DMF. The direct or indirect (via EET) population of Pc moiety is followed by the hole transfer to the charge-separated state in 4 (in DMF) and in 5 (in DMF and toluene). At the same time, it was found that CT is energetically unfavorable for the triad 4 in toluene upon excitation of the Pc-part. The charge-recombination in DMF occurs very fast with a decay time of 40 and 30 ps for 4 and 5, respectively, whereas toluene stabilizes the charge-separated state, prolonging the lifetime to 4.5 and 1.7 ns, respectively. PMID- 19809676 TI - Electrochemical investigation of Mn4O4-cubane water-oxidizing clusters. AB - High valence states in manganese clusters are a key feature of the function of one of the most important catalysts found in nature, the water-oxidizing complex of photosystem II. We describe a detailed electrochemical investigation of two bio-inspired manganese-oxo complexes, [Mn(4)O(4)L(6)] (L = diphenylphosphinate (1) and bis(p-methoxyphenyl)phosphinate (2)), in solution, attached to an electrode surface and suspended within a Nafion film. These complexes contain a cubic [Mn(4)O(4)](6+) core stabilized by phosphinate ligands. They have previously been shown to be active and durable photocatalysts for the oxidation of water to dioxygen. A comparison of catalytic photocurrent generated by films deposited by two methods of electrode immobilization reveals that doping of the catalyst in Nafion results in higher photocurrent than was observed for a solid layer of cubane on an electrode surface. In dichloromethane solution, and under conditions of cyclic voltammetry, the one-electron oxidation processes 1/1(+) and 2/2(+) were found to be reversible and quasi-reversible, respectively. Some decomposition of 1(+) and 2(+) was detected on the longer timescale of bulk electrolysis. Both compounds also undergo a two-electron, chemically irreversible reduction in dichloromethane, with a mechanism that is dependent on scan rate and influenced by the presence of a proton donor. When immersed in aqueous electrolyte, the reduction process exhibits a limited level of chemical reversibility. These data provide insights into the catalytic operation of these molecules during photo-assisted electrolysis of water and highlight the importance of the strongly electron-donating ligand environment about the manganese ions in the ability of the cubanes to photocatalyze water oxidation at low overpotentials. PMID- 19809677 TI - Photo-controlled conformation-assisted permanent optical storage device employing a polymer network liquid crystal. AB - A new type of optical storage device is described employing a material consisting of a host nematic liquid crystal, a photoisomerisable azobenzene component and a photopolymerizable monomer. The principle of image storing involves selectively controlling the birefringence of the medium immediately prior to photopolymerization of the monomer. We show that photoisomerisation driven nematic to isotropic transition can be employed to achieve this through proper timing of the reverse isomerization of the azobenzene compound before the nematic director fluctuations get quenched. It is also suggested that grey-shades can be created in this device using the recently discovered phenomenon of electric-field acceleration of reverse isomerisation. PMID- 19809678 TI - Characterization of two lactones in liquid phase: an experimental and computational approach. AB - In this work, an experimental and computational study on the properties and molecular-level liquid structure of gamma-butyrolactone and gamma-valerolactone is reported. These fluids are selected because of their possible use as alternative green solvents considering their favorable environmental and toxicological profiles. The experimental study was carried out measuring a selected collection of relevant thermophysical properties, at ambient pressure as a function of temperature, together with a detailed spectroscopic study. The reported set of experimental thermophysical properties is valuable for process design purposes and because of their relationships with microscopic fluids' structure. Spectroscopic studies were carried out using attenuated total reflection infrared spectroscopy (ATR-FTIR), solvatochromic studies and microwave dielectric relaxation spectroscopy (DRS) measurements; all the spectroscopic studies were performed as a function of temperature. Moreover, density functional theory (DFT) and classical molecular dynamics simulations (MD) were used to obtain a detailed picture of the intermolecular interactions within the fluid, at short and long ranges, and of other relevant features leading to the liquid structure of the studied fluids. The whole study points to a fluids' picture in which, in spite of the absence of specific interactions such as hydrogen bonding, a remarkable ordering appears rising from the dipolar interactions and from the shapes of the involved molecules. PMID- 19809679 TI - Modeling of the Zn2+ binding in the 1-16 region of the amyloid beta peptide involved in Alzheimer's disease. AB - Zinc ions are found at mM concentration in amyloid plaques of Alzheimer's disease and the role of zinc in protein oligomerization is the object of intense investigations. As an in vitro model for studying interactions between Zn(2+) and the Abeta peptide, that is the main component of plaques, the N- and C-termini protected Abeta(1-16) fragment has been chosen because reliable spectroscopic studies in water solution are possible due to the low propensity for oligomerization at pH approximately 6.5, and because all the Zn binding sites of Abeta have been identified in the 1-16 region. In this work we present the results of first principle simulations of several initial models of Zn-Abeta(1 16) complexes. The NMR results about the same system, where His 6, 13, 14 and Glu 11 side-chains coordinate the Zn ion, are strongly supported by these models. Coordination of Asp 1 to Zn drives the complex towards the expulsion of one of initially bonded His side-chains. Coordination of Tyr 10 to Zn is possible only when Tyr 10 is deprotonated. The interplay between physico-chemical properties of the Abeta ligand and the Zn coordination is discussed. PMID- 19809680 TI - Solvation shell structure of cyclooctylpyranone in water solvent and its comparative structure, dynamics and dipole moment in HIV protease. AB - We have investigated the solvation structure for cyclooctylpyranone (COP) in water solvent using force-field molecular dynamics (MD) and Car-Parrinello mixed quantum mechanics-molecular mechanics (CPMD) calculations. The MD calculations show that in water solvent COP can exist in two conformational states which differ with respect to the relative orientations of the three rings, namely phenyl, pyranone and cyclooctane. We report the existence of strong orientational preference for the water molecule in the first solvation shell and the orientational preference disappears for solvent molecules beyond the first solvation shell. In order to investigate the confinement effect on the structure, dynamics, charge distribution and dipole moment of COP, we have carried out MD and CPMD calculations for COP within HIV type-1 protease (PR). Interestingly, we do not see any conformational transitions for COP within the protein cavity and it remains as a single conformer. We do see a remarkable effect of confinement on few other torsional degrees of freedom such as gg to tg conformational shift for the propyl group of COP. However, the methyl group rotational dynamics remains similar in the water solvent and in the protein environment. Also, within the protein cavity, the COP molecule is more polarized when compared to water solvent. Static ab initio electronic structure calculations were performed on the COP molecule with varying torsional angle in order to investigate the angle dependence of the molecular volume and energy. PMID- 19809681 TI - Hydrophobic coating- and surface active solvent-mediated self-assembly of charged gold and silver nanoparticles at water-air and water-oil interfaces. AB - We report self-assembly of charge-stabilized gold and silver nanoparticles at water-air and water-oil interfaces, via manipulation of the interactions between the interfaces and the adsorbing nanoparticles. Nanoparticle adsorption from bulk colloids to an interface is an energy-favored, but finite sorption barrier restrained (kinetics-controlled) process. Consequently, to successfully mediate self-assembly of nanoparticles, the finite sorption barrier should be decreased. That can be accomplished by manipulating its three controlling forces: the repulsive electrostatic force, the repulsive van der Waals force, and the attractive hydrophobic force between the interface and the adsorbing nanoparticles. It was found that hydrophobic coatings change nanoparticle hydrophobicity and greatly increase the attractive hydrophobic force. Surface active organic solvents (methanol, ethanol, isopropanol, and acetone) decrease the attractive hydrophobic force to some extent. However, they decrease the repulsive electrostatic force to a larger extent, via a "charge dilution" mechanism, due to their positive adsorption at the charged water-air and water oil interfaces. Hydrophobic coatings and organic solvents consequently decrease the sorption barrier, facilitate nanoparticles overcoming the sorption barrier, and mediate the self-assembly of nanoparticles. PMID- 19809682 TI - Substrate effect on supramolecular self-assembly: from semiconductors to metals. AB - Terephthalic acid (TPA) deposited on Si(111)-7 x 7, Si(111)-square root 3 x square root 3-Ag and Ag(111) has been studied as a model system to understand how much passivated semiconductor surfaces differ from semiconductor and metal surfaces in respect of supramolecular self assembly. By scanning tunneling microscopy it is found that TPA molecules do not form any ordered supramolecular structure on the pristine semiconductor surface, due to a strong molecule substrate interaction. On the contrary, TPA has a weaker interaction with Si(111) square root 3 x square root 3-Ag, leading to the formation of an ordered supramolecular layer stabilized by carboxyl hydrogen bonds. These structures are very similar to the supramolecular layer of TPA formed on Ag(111), indicating that the two substrates behave similarly for what concerns the adsorption of functional organic molecules. However, the deposition of Fe on the TPA layers on Si(111)-square root 3 x square root 3-Ag does not induce the formation of two dimensional metal-organic frameworks which, on the contrary, readily develop on Ag(111). Possible origins of this difference are discussed. PMID- 19809683 TI - In situ ESR-UV-Vis-NIR spectroelectrochemical study of the p-doping of poly[2-(3 thienyl)ethyl acetate] and its hydrolyzed derivatives. AB - The electrochemical oxidation of the chemically prepared polymer poly[2-(3 thienyl)ethyl acetate] (PTEtAc), its partially hydrolyzed derivative PTEtAcOH and the fully hydrolyzed compound poly[2-(3-thienyl)ethanol] (PTEtOH) was studied by in situ electron spin resonance (ESR)/UV-Vis-NIR spectroelectrochemistry. The spectroelectrochemical response of these films on ITO substrates was analyzed with respect to the influence of the functionalized alkyl side chain on polymer doping. The simultaneous use of both electron spin resonance and UV-Vis-NIR spectroscopy allows the analysis of the nature, extent and stability of the charge carriers electrogenerated during p-doping. It was found that PTEtAc has a higher capacity for charged species due to the flexibility of the longer side chains making the redox states more stable at different doping levels. At low doping levels the charged states are dominated by polaronic species while at high doping levels bipolarons and diamagnetic polaron pairs are formed. The presence of the OH groups in the polymer side chains of the hydrolyzed derivatives favors hydrogen bonds. These interactions by hydrogen bonding fix the conjugated chains thus making a charge-discharge reaction more difficult. At high doping levels the hydrolyzed polymers favor the formation of polaron pairs. PMID- 19809684 TI - Theoretical investigation on the influence of temperature and crystallographic orientation on the breaking behavior of copper nanowire. AB - In this paper, molecular dynamics simulations have been conducted to study the mechanical stretching of copper nanowires which will finally lead to the formation of suspended liner atomic chains. A total of 2700 samples have been investigated to achieve a comprehensive understanding of the influence of temperature and orientation on the formation of linear atomic chains. Our results prove that linear atomic chains do exist for [100], [111] and [110] crystallographic directions. Stretching along the [111] direction exhibits a higher probability in forming the two-atom contact than that along the [110] and [100] directions. However, for longer linear atomic chains, there emerges a reversed trend. In addition, increasing temperature may decrease the formation probability for stretching along [111] and [110] directions, but this influence is less obvious for that along the [100] direction. PMID- 19809685 TI - Mechanisms of water infiltration into conical hydrophobic nanopores. AB - Fluid channels with inclined solid walls (e.g. cone- and slit-shaped pores) have wide and promising applications in micro- and nano-engineering and science. In this paper, we use molecular dynamics (MD) simulations to investigate the mechanisms of water infiltration (adsorption) into cone-shaped nanopores made of a hydrophobic graphene sheet. When the apex angle is relatively small, an external pressure is required to initiate infiltration and the pressure should keep increasing in order to further advance the water front inside the nanopore. By enlarging the apex angle, the pressure required for sustaining infiltration can be effectively lowered. When the apex angle is sufficiently large, under ambient condition water can spontaneously infiltrate to a certain depth of the nanopore, after which an external pressure is still required to infiltrate more water molecules. The unusual involvement of both spontaneous and pressure assisted infiltration mechanisms in the case of blunt nanocones, as well as other unique nanofluid characteristics, is explained by the Young's relation enriched with the size effects of surface tension and contact angle in the nanoscale confinement. PMID- 19809686 TI - Periodic DFT modeling of bulk and surface properties of MgCl2. AB - MgCl(2) is the preferred support for the industrial Ziegler-Natta catalysts, and is believed to act as a template for the epitactic chemisorption of the active Ti species. As the first step of a thorough computational modeling of these systems, we studied the bulk and surface structure of the ordered alpha and beta phases of MgCl(2) by means of periodic DFT (B3LYP) methods using localized basis sets. The layer structure of both phases was reproduced satisfactorily with the inclusion of a (small) empirical dispersion correction ("DFT-D") as a practical method to describe the attraction between the layers. Surface models were studied on slabs with adequate thickness. It appears that various surfaces exposing 5-coordinated Mg are very similar in energy and are the lowest non-trivial surfaces. Cuts exposing 4-coordinated Mg are significantly less stable; both kinetic and equilibrium models of crystal growth indicate that they should normally not be formed to a significant extent. "Nano-ribbons" of single, flat chains of MgCl(2), sometimes proposed as components of the disordered delta phase, were also evaluated, but are predicted to be unstable to rearrangement. Implications for the role of MgCl(2) as catalyst support are discussed. PMID- 19809694 TI - Environmental monitoring: a changing challenge. PMID- 19809695 TI - A new look at low-energy nuclear reaction research. AB - This paper presents a new look at low-energy nuclear reaction research, a field that has developed from one of the most controversial subjects in science, "cold fusion." Early in the history of this controversy, beginning in 1989, a strong polarity existed; many scientists fiercely defended the claim of new physical effects as well as a new process in which like-charged atomic nuclei overcome the Coulomb barrier at normal temperatures and pressures. Many other scientists considered the entire collection of physical observations-along with the hypothesis of a "cold fusion"--entirely a mistake. Twenty years later, some people who had dismissed the field in its entirety are considering the validity of at least some of the reported experimental phenomena. As well, some researchers in the field are wondering whether the underlying phenomena may be not a fusion process but a neutron capture/absorption process. In 2002, a related tabletop form of thermonuclear fusion was discovered in the field of acoustic inertial confinement fusion. We briefly review some of this work, as well. PMID- 19809696 TI - Determination of trace element concentrations in natural freshwaters: how low is "low", and how low do we need to go? AB - There is an on-going need for reliable concentration data for trace elements in natural freshwaters, including soil solutions and groundwaters, surface waters (wetlands, streams, rivers, and lakes), precipitation (fog, rain, and snow), and drinking water (including natural spring water as well as bottled water and tap water). Some of the trace elements of interest may be present in these waters at elevated concentrations due either to natural processes such as mineral weathering (e.g. As and U in groundwater), or because of human activities (e.g. atmospheric contamination of snow with Pb and Sb). PMID- 19809697 TI - Perspective on element speciation. AB - During the second half of the 20th century it became evident that trace elements play a major role whenever biological activities, environmental chemistry, or material characteristics are discussed and the determination of trace elements has therefore gained outstanding importance in environmental and life sciences. Elements, even when present at minimal concentrations in biological and environmental matrices, can, in fact, exert fundamental influence on ecosystems and the vital functions of organisms. The study of, for example, pathophysiological processes in the human body requires the determination of elements down to the ng kg(-1) range. However, organic and inorganic matrix compounds at much higher concentration levels in a sample than trace elements make the determination of trace elements often rather difficult. PMID- 19809698 TI - Foreword: JEM spotlight: Environmental monitoring of airborne nanoparticles. PMID- 19809699 TI - JEM spotlight: Environmental monitoring of airborne nanoparticles. AB - The aim of this work was to review the existing instrumental methods to monitor airborne nanoparticles in different types of indoor and outdoor environments in order to detect their presence and to characterise their properties. Firstly the terminology and definitions used in this field are discussed, which is followed by a review of the methods to measure particle physical characteristics including number, concentration, size distribution and surface area. An extensive discussion is provided on the direct methods for particle elemental composition measurements, as well as on indirect methods providing information on particle volatility and solubility, and thus in turn on volatile and semivolatile compounds of which the particle is composed. A brief summary of broader considerations related to nanoparticle monitoring in different environments concludes the paper. PMID- 19809700 TI - Setting the limits for engineered nanoparticles in European surface waters - are current approaches appropriate? AB - The current widespread and diverse use of engineered nanoparticles (ENPs) in consumer products is expected to result in direct and indirect emissions to the aquatic environment. In Europe, the Water Framework Directive (WFD) is responsible for maintaining a good chemical and ecological status of surface waters. This review provides an insight to the likely scenarios if an engineered nanoparticle should be considered a "priority substance" in the WFD. Through a brief literature review of the aquatic toxicity of carbon- and metal-containing ENPs and in light of principles to establish environmental quality standards, we conclude that it is impossible to set limit values for ENPs in surface waters now and in the foreseeable future. This is due to the extensive lack of knowledge not only in relation to unknown toxic effects, degradability, and bioaccumulation of ENPs in the aquatic environment, but also due to the questionable validity of test systems and methods to establish environmental quality standards (EQS). From this, we also suggest some paths to follow to compensate for these knowledge gaps which include environmental monitoring and a closer look at the use of ENPs in consumer products. PMID- 19809701 TI - Sensors as tools for quantitation, nanotoxicity and nanomonitoring assessment of engineered nanomaterials. AB - The discovery of fullerenes in 1985 has ushered in an explosive growth in the applications of engineered nanomaterials and consumer products. Nanotechnology and engineered nanomaterials (ENMs) are being incorporated into a range of commercial products such as consumer electronics, cosmetics, imaging and sensors. Nanomaterials offer new possibilities for the development of novel sensing and monitoring technologies. Nanosensors can be classified under two main categories: (i) Nanotechnology-enabled sensors or sensors that are themselves nanoscale or have nanoscale materials or components, and (ii) Nanoproperty-quantifiable sensors or sensors that are used to measure nanoscale properties. The first category can eventually result in lower material cost, reduced weight and energy consumption. The second category can enhance our understanding of the potential toxic effects of emerging pollutants from nanomaterials including fullerenes, dendrimers, and carbon nanotubes. Despite the enormous literatures and reviews on Category I sensors, there are few sensors to measure nanoscale properties or sensors belonging to Category II. This class of nanosensors is an area of critical interest to nanotoxicology, detection and risk assessment, as well as for monitoring of environmental and/or biological exposure. This article discusses emerging fields of nanotoxicology and nanomonitoring including the challenges of characterizing engineered nanomaterials and the potentials of combining existing analytical techniques with conventional cytotoxicity methods. Two case studies are provided for development of Category II nanosensors for fullerene nanoparticles and quantum dots. One highlights the uniqueness of a portable, dissolved oxygen electrochemical sensor arrays capable of detecting the ENMs as well as provide rapid nanotoxicological information. This review has shown that addressing the complex and critical issues surrounding the environmental transformation and toxicity of ENMs must be accompanied by the creation of new approaches or further developments of existing instrumentation. PMID- 19809702 TI - A review of oil-suspended particulate matter aggregation--a natural process of cleansing spilled oil in the aquatic environment. AB - It has been acknowledged that following an oil spill in coastal areas where suspended particulate matter (SPM) is rich, aggregation between oil and SPM can be naturally formed. This kind of aggregation product is termed as oil-SPM aggregates (OSAs). Because OSAs are not as sticky to the shorelines as crude oil and the oil-water contact area is greatly increased due to the formation of OSAs, both oil dispersion into the water body and oil biodegration would be significantly enhanced. In this review article, the authors (1) describe in detail the mechanism of OSA formation and controlling parameters which can influence OSA formation (the parameters discussed include the oil nature and properties, sediment types and concentrations, and the environmental factors such as salinity, temperature and mixing energy); (2) briefly review qualitative and quantitative methods used for characterization of OSA formation (two main methods used for the OSA characterization are the UV epi-fluorescence microscopy and gas chromatography equipped with flame ionization detector (GC-FID); (3) elucidate the applications of OSA formation in oil spill response strategies including natural attenuation, sediment relocation, and sediment mixing; and (4) discuss research needs in the future which would further improve our understanding of OSA formation and move towards the development of adequate oil behaviour models. PMID- 19809703 TI - Soil macronutrient sensing for precision agriculture. AB - Accurate measurements of soil macronutrients (i.e., nitrogen, phosphorus, and potassium) are needed for efficient agricultural production, including site specific crop management (SSCM), where fertilizer nutrient application rates are adjusted spatially based on local requirements. Rapid, non-destructive quantification of soil properties, including nutrient levels, has been possible with optical diffuse reflectance sensing. Another approach, electrochemical sensing based on ion-selective electrodes or ion-selective field effect transistors, has been recognized as useful in real-time analysis because of its simplicity, portability, rapid response, and ability to directly measure the analyte with a wide range of sensitivity. Current sensor developments and related technologies that are applicable to the measurement of soil macronutrients for SSCM are comprehensively reviewed. Examples of optical and electrochemical sensors applied in soil analyses are given, while advantages and obstacles to their adoption are discussed. It is proposed that on-the-go vehicle-based sensing systems have potential for efficiently and rapidly characterizing variability of soil macronutrients within a field. PMID- 19809704 TI - Is it necessary to raise awareness about technologically enhanced naturally occurring radioactive materials? AB - Since radiation risks are usually considered to be related to nuclear energy, the majority of research on radiation protection has focused on artificial radionuclides in radioactive wastes, spent nuclear fuel or global fallout caused by A-bomb tests and nuclear power plant failures. Far less attention has been paid to the radiation risk caused by exposure to ionizing radiation originating from natural radioactivity enhanced due to human activity, despite the fact that technologically enhanced naturally occurring radioactive materials are common in many branches of the non-nuclear industry. They differ significantly from "classical" nuclear materials and usually look like other industrial waste. The derived radiation risk is usually associated with risk caused by other pollutants and can not be controlled by applying rules designed for pure radioactive waste. Existing data have pointed out a strong need to take into account the non-nuclear industry where materials containing enhanced natural radioactivity occur as a special case of radiation risk and enclose them in the frame of the formal control. But up to now there are no reasonable and clear regulations in this matter. As a result, the non-nuclear industries of concern are not aware of problems connected with natural radioactivity or they would expect negative consequences in the case of implementing radiation protection measures. The modification of widely comprehended environmental legislation with requirements taken from radiation protection seems to be the first step to solve this problem and raise awareness about enhanced natural radioactivity for all stakeholders of concern. PMID- 19809705 TI - Foreword: JEM spotlight: Monitoring the treatment efficiency of a full scale ozonation on a sewage treatment plant with a mode-of-action based test battery. PMID- 19809706 TI - JEM spotlight: Monitoring the treatment efficiency of a full scale ozonation on a sewage treatment plant with a mode-of-action based test battery. AB - Tertiary treatment of wastewater with ozone is a promising technique for removing residual micropollutants that remain after secondary biological treatment. We monitored the performance of a full-scale ozonation reactor on a sewage treatment plant in Switzerland with a screening battery of bioassays. Six toxicity endpoints were selected that covered non-specific toxicity, as well as selected receptor-mediated modes of action and reactive toxicity. Non-specific toxicity was assessed with two bioassays, the bioluminescence inhibition of the marine luminescent bacterium Vibrio Fischeri and the growth inhibition of the green algae Pseudokirchneriella subcapitata. Treatment efficiency was around 90% for the secondary treatment, but only 65% and 76% for the ozonation step in the two non-specific endpoints, respectively. This finding is consistent with this type of oxidation reaction because ozone only modifies the organic molecules but does not mineralize them fully leaving residual toxicity of the transformation products. In contrast, the specific receptor-mediated endpoints of inhibition of photosystem II in algae and estrogenicity were largely reduced by ozonation. While compounds inhibiting photosynthesis proved to be rather recalcitrant toward biological treatment with only 47% removal, an additional 86% removal by ozonation yielded an overall treatment efficiency in the entire treatment chain of 89%. The effect on estrogenicity, quantified with the yeast estrogen screen, was even more significant: A treatment efficiency of 95% in the secondary treatment, 86% during ozonation plus a small effect by biological sand filtration yielded an overall treatment efficiency of 99.5%. Insecticides that inhibit acetylcholinesterase were fairly resistant to degradation, but an overall treatment efficiency of 91% was achieved in two steps: 72% in biological treatment and 60% during ozonation. Finally, no significant genotoxicity was observed with the umuC test after ozonation, while the influent showed a genotoxic response when it was enriched by a factor of 15 to 60. Treatment efficiency increased with the ozone dose and remained virtually unchanged over ozone doses above 500 g ozone per kg dissolved organic carbon. The reduction of toxicity can be rationalized by the chemical oxidation processes likely to occur for each group of chemicals that are typical for a given mode of toxic action. For comparison, tertiary treatment with powdered activated carbon was also evaluated, which poses a viable alternative to ozonation with respect to removal of micropollutants. PMID- 19809707 TI - Foreword: JEM spotlight: Fungi, mycotoxins and microbial volatile organic compounds in mouldy interiors from water-damaged buildings. PMID- 19809708 TI - JEM Spotlight: Fungi, mycotoxins and microbial volatile organic compounds in mouldy interiors from water-damaged buildings. AB - Concerns have been raised about exposure to mycotoxin producing fungi and the microbial volatile organic compounds (MVOCs) they produce in indoor environments. Therefore, the presence of fungi and mycotoxins was investigated in 99 samples (air, dust, wallpaper, mycelium or silicone) collected in the mouldy interiors of seven water-damaged buildings. In addition, volatile organic compounds (VOCs) were sampled. The mycotoxins were analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (20 target mycotoxins) and quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). Morphological and molecular identifications of fungi were performed. Of the 99 samples analysed, the presence of one or more mycotoxins was shown in 62 samples by means of LC-MS/MS analysis. The mycotoxins found were mainly roquefortine C, chaetoglobosin A and sterigmatocystin but also roridin E, ochratoxin A, aflatoxin B(1) and aflatoxin B(2) were detected. Q-TOF MS analysis elucidated the possible occurrence of another 42 different fungal metabolites. In general, the fungi identified matched well with the mycotoxins detected. The most common fungal species found were Penicillium chrysogenum, Aspergillus versicolor (group), Chaetomium spp. and Cladosporium spp. In addition, one hundred and seventeen (M)VOCs were identified, especially linear alkanes (C(9)-C(17)), aldehydes, aromatic compounds and monoterpenes. PMID- 19809709 TI - Field calibration of polyurethane foam disk passive air samplers for PBDEs. AB - A field study was performed to derive uptake rates of airborne polybrominated diphenyl ethers (PBDEs) to polyurethane foam (PUF) disk passive air samplers (PAS) and to investigate the influence of deployment location and device design. Data are presented on the gas-particle partitioning of PBDEs, since atmospheric phase distribution was considered to be a variable which could affect sampler performance. Uptake rates for these compounds were similar to those derived previously for other classes of persistent organic pollutants (POPs) (approximately 2-6 m(3)/day), with rates higher for the higher brominated species. Whilst other compound classes (e.g. polychlorinated biphenyls) are predominantly present in the air in the gas phase, heavier PBDEs have an association with particulates in the atmosphere at ambient temperatures. In this study, the PUF disk PAS therefore sampled PBDEs present in the gas phase and on fine aerosols with a similar sampling efficiency to those which are predominantly gas phase compounds. Compounds which are exclusively on particles are sampled less efficiently. A comparison of the three most commonly used PUF deployment configurations, used by different research groups, indicated little difference in uptake rates. The ranges of derived air concentrations for BDE-47, -99, and -183 between three sampler designs were 7.5-9.8, 7.4-12.4, and 4.7-6.6 pg/m(3), respectively. This suggests the robustness of this sampler in comparisons between regional and global campaigns where these three designs are employed. PMID- 19809710 TI - Pesticide sequestration in passive samplers (SPMDs): considerations for deployment time, biofouling, and stream flow in a tropical watershed. AB - Semi-permeable membrane devices (SPMDs) provide an informative and cost-effective approach for monitoring contaminants in remote tropical streams. Estimation and interpretation of contaminant concentrations in streams derived from SPMDs can vary based on a number of environmental factors, including stream flow, biofouling, and deployment time. In three one-month long trials, SPMDs were concurrently deployed for 4, 15, and 28 days at three stream sites in an extensive agricultural area of southeastern Costa Rica. Water, bottom sediment, and suspended solids grab samples were also collected and several environmental variables were monitored at corresponding time intervals during each month-long study period. At all three sites, SPMD concentrations of the widely used insecticide chlorpyrifos increased with deployment time, with no relationship between SPMD biofouling and pesticide sequestration. Differences in SPMD chlorpyrifos sequestration among sites are likely due to differences in stream chlorpyrifos concentration rather than differences in SPMD sampling rates. The longer exposure period of SPMDs allowed for the detection of lower concentrations of chlorpyrifos, terbufos, and difenoconazole compared to water grab samples. In addition to the use of appropriate performance reference compounds (PRCs), other environmental variables such as stream turbidity, flow regime, stream morphology, and knowledge of pesticide application methods are important considerations for optimizing SPMD deployment and data interpretation in tropical regions. PMID- 19809711 TI - Distribution and relationships of trace metals in the isopod Saduria entomon and adjacent bottom sediments in the southern Baltic. AB - The concentrations of Ag, Cd, Co, Cr, Cu, Fe, Ni, Pb, Mn and Zn in Saduria entomon and adjacent bottom sediments from the southern Baltic were determined by FAAS. In order to estimate the strength of correlations between accumulated elements in these crustaceans and surficial sediment, bioaccumulation factors (BAFs) were calculated. The results of factor analysis (FA) and the Kruskal Wallis analysis of variance (ANOVA) clearly indicate geographical differences between the concentrations of these elements. Cd, Co, Fe, Ni, Pb and Zn levels were higher in S. entomon from the Gulf of Gdansk, whereas Cr and Mn levels were higher in the crustaceans inhabiting open Baltic waters. The concentrations of Ag and Cu were comparable in both regions. There was a tendency for metal concentrations to distinguish organisms inhabiting the muddy bottom from those living in sandy sediments. The granulometric composition of the sediment appears to influence trace metal bioavailability. The results show that S. entomon could be a valuable sentinel organism for biomonitoring heavy metal contamination in the southern Baltic. PMID- 19809717 TI - A tribute to Frederick Nye Tebbe. Lewis acid stabilized alkylidyne, alkylidene, and imides of 3d early transition metals. AB - Frederick Nye Tebbe distanced himself from the spotlight despite increasing appreciation of his discoveries by the scientific world. Although his research accomplishments are numerous and significant, perhaps his most highly recognized contribution is the reagent that bears his name, Cp2Ti(mu2-CH2)(mu2-Cl)Al(CH3)2: an inspiring molecule to the area of olefin metathesis and methylidene transfer. Masking its potential via Lewis acid stabilization, Tebbe tamed the titanium methylidene moiety, leading many other scientists to exploit the wide-ranging reactivity of such a unit, including olefin metathesis, methylenation, and metallacyclobutane formation, among others. In this perspective, we pay tribute to the life and chemistry of the man behind the masked methylidene and the research progeny spawned by his discovery, focusing on the Lewis acid stabilization of multiply-bonded ligands of the early metals such as Sc and Ti. PMID- 19809718 TI - Palladium complexes containing novel cyclic selenium imides. AB - The first metal complexes of cyclic selenium(II) imide ligands, [PdCl2{Se,Se' Se4(NtBu)3}] and [PdCl2{Se,Se'-Se4(NtBu)4}], which are obtained from the reaction of Se(NtBu)2 with [PdCl2(NCPh)2], contain the novel Se-N heterocycles Se4(NtBu)3 and Se4(NtBu)4. PMID- 19809719 TI - Controlling Cu...Cu distances using halides: (8-phenylthionaphth-1 yl)diphenylphosphine copper halide dimers. AB - In the isomorphous binuclear Cu2X2L2 systems (L = (8-phenylthionaphth-1 yl)diphenylphosphine the Cu...Cu separation is reduced as the halide size increases. PMID- 19809720 TI - Environmentally benign synthesis of virus-templated, monodisperse, iron-platinum nanoparticles. AB - The use of an engineered variant of a plant virus, Cowpea mosaic virus (CPMV), as a template for directed mineralization provides an environmentally benign route to monodisperse iron-platinum nanoparticles of approximately 30 nm diameter. PMID- 19809721 TI - The mechanism of quenching of the lanthanide excited state for optical probes using sensitised emission. AB - The mechanism of quenching of the excited state of terbium and europium complexes by electron-rich reductants occurs by formation of an exciplex involving the triplet excited state of the sensitising chromophore. PMID- 19809722 TI - C2-symmetric S/C/S ligands based on N-heterocyclic carbenes: a new ligand architecture for asymmetric catalysis. AB - Neutral, C2-symmetric S/C/S ligands based on N-heterocyclic carbenes and thioether functionalities were incorporated into transition metal complexes characterised by two direct metal-stereogenic sulfur bonds. This new ligand design was applied to 1,3-dipolar cycloadditions as the first example of the use silver of carbenes in asymmetric catalysis. PMID- 19809723 TI - A monometallic tri-spin single-molecule magnet based on rare earth radicals. AB - A mononuclear tri-spin single-molecule magnet based on the rare earth radical [Tb(hfac)3(NITPhOEt)2] (NITPhOEt = 4'-ethoxy-phenyl-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide) has been synthesized, structurally characterized and the alternating current signals show a slow relaxation of magnetization and frequency-dependent signals. PMID- 19809724 TI - Vicinal diphosphoniums: electrostatic repulsion under covalent constraint. AB - A series of five vicinal bis(alkyl-triarylphosphoniums) derived from o bis(diphenyl-phosphino)benzene (o-dppb) is described. Each of them have been prepared by specific methods, and their formal electrostatic and possible van der Waals strain is compared through the P+...P+ distances in the crystal state. According to X-ray diffraction analyses, while the conformations of the dimethyl dication is C2 symmetric, the conformation of alka-1,n-diyl-diphosphoniums (n = 1, 2, 3) is pseudo-Cs symmetric. The solution structure of the dimethyldiphosphonium 2 was studied by NMR techniques after metathesis of the triflate counterions with enantiomerically pure X-PHAT chiral anions (X = TRIS, BIN). [(Delta)-BINPHAT] was indirectly shown to discriminate between the enantiomers of the diphosphonium, but no effective enantio-differentiation could be measured down to -70 degrees C in CD2Cl2. When associated to [(Delta) TRISPHAT] counterions, progressive cleavage of the diphosphonium afforded the corresponding methyl-triphenyl-monophosphonium salt. In the bridged series, the diphosphoniacyclopentene dication was found to be stable in spite of the very short transannular P+...P+ distance (2.83 A). The naturally more relaxed homologous diphosphoniacyclohexene dication (P+...P+ = 3.28 A) was found to be flexible in solution, as indicated by the equivalence of the four protons of the +P-CH2CH2-P+ bridge from 20 degrees C to -80 degrees C in acetone-d6. The two diphosphoniacycloheptene dications exhibit no P+...P+ steric compression (ca 3.64 A), just as the dimethyl dication (3.70 A). In solution, the seven-membered ring of the +P-CH2CR2CH2-P+ dications (R = H, Me) remains rigidly Cs symmetric: no interconversion of the pseudo-axial and pseudo-equatorial R groups is evidenced at the NMR time scale at -95 degrees C (coalescence at -20 degrees C for R = H, at -65 degrees C for R = Me). According to DFT calculations at the B3PW91/6-31G** level in 1,2-dichloroethane (DCE: PCM, epsilon = 10.36), the lowest energy conformation is indeed Cs-symmetric. However a quasi-isoenergetic C2-symmetric conformation which is the most stable in the gas phase is also found, and is thus a likely intermediate in the observed Cs <==> Cs interconversion. This +P CH2CMe2CH2-P+ diphosphonium was obtained by dialkylation of o-dppb with 2,2 dimethylpropan-1,3-diyl bistriflate in DCE at 80 degrees C. The use of this solvent was the key for the synthesis of this dication, which was however competitively produced along with some +P-CH2CH2-P+ dication. The latter was also obtained by a totally different route, by heating a sulfinylethyl monophosphonium salt of o-dppb in the presence of the cationic complex [Rh(cod)2][PF6]. A mechanism for this peculiar process is proposed. PMID- 19809725 TI - Binary and ternary lanthanide centered hybrid polymeric materials: coordination bonding construction, characterization, microstructure and photoluminescence. AB - A functional molecular bridge (named as HBA-TEPIC) (HBA = 1,4-hydroxybenzoic acid, TEPIC = 3-(triethoxysilyl)-propyl-isocyanate) was achieved through the hydrogen transfer nucleophilic addition reaction. Firstly, the molecular precursors coordinate to lanthanide ions (Eu3+ and Tb3+) and then form the covalently bonded Si-O network to obtain the polymeric hybrid material (HBA-TEPIC RE). Secondly, the synthesized polymers PMMA and PMAALM and commercial PVP (polyvinylpyrrolidone) were further introduced into the inorganic network to obtain hybrids with the organic chains and inorganic networks together (HBA-TEPIC RE-PVP/PMAA/PMAALM). The regular microstructure indicates that a self-assembly system exists and both lanthanide ions and polymeric chains have an influence on the growth tendency of the hybrids. The results of the luminescent properties of the hybrids prove that the HBA-TEPIC-Eu-PVP hybrid represents the longest lifetime and highest quantum efficiency. PMID- 19809726 TI - Synthesis and visible light photocatalytic property of polyhedron-shaped AgNbO3. AB - Polyhedron-shaped AgNbO3 photocatalysts were synthesized by solvothermal and liquid-solid methods. Their photocatalytic properties were evaluated from the photocatalytic O2 evolution under visible light irradiation. The polyhedron shaped AgNbO3 was induced to grow by shaped silver particles followed by the free growth model. The photocatalytic results indicate that the polyhedron-shaped morphology is favourable for the photocatalytic O2 evolution under visible light irradiation in comparison with the spherical one. Furthermore, the Cu doping on the surface would enhance the visible light photocatalytic activity significantly. PMID- 19809727 TI - Apoptosis and anti-tumour activities of manganese(III)-salen and -salphen complexes. AB - We analyzed the apoptosis and anti-tumour activities of several Mn(III)-salen and -salphen complexes (1-14) towards three different cultured human cancer and non cancer cells. We demonstrated that most of the Mn(III)-salen and -salphen complexes affect cell viability and induce apoptosis in MCF7 cells. Biochemically active Mn(III)-salen and -salphen complexes induced nuclear fragmentation and release of cytochrome c from the mitochondria to cytosol indicating involvement of mitochondrial pathway of apoptosis. The nature and position of the substituents and the bridging group on the salen ligands play crucial roles in determining the apoptotic activities of Mn(III)-salen and -salphen complexes. The IC50 values for the active Mn(III)-salen complexes ranged between 12 and 55 microM. For Mn(III)-salen complexes with ethylenediamine bridges, methoxy substituted complexes were more active than the corresponding hydroxy derivatives. However, this correlation does not hold when the bridging group was changed from ethylenediamine to o-phenylenediamine. Importantly, several Mn(III) salen and -salphen complexes showed about 2-3 fold selectivity toward cancer cells such as MCF7 (breast cancer), and CCL228 (colon cancer) over a normal non malignant cell MCF10 (breast epithelial cells) indicating their potential application towards novel anti-tumour therapy. PMID- 19809728 TI - Effect of Lewis acid on the structure of a diiron dithiolate complex based on the active site of [FeFe]-hydrogenase assessed by density functional theory. AB - The effect of Lewis acid on the structure and H2 productivity of a diiron dithiolate complex was investigated by using density functional theory (DFT) calculations. When a model molecule of [(CH3SH)(CO)2Fe(p)(mu SCH2NHCH2S)Fe(d)(CO)3] was geometrically optimized, two isomers were found: one is the unrotated structure (1) with no ligand between two Fe atoms and the other is the rotated structure (1*) with one CO ligand between two Fe atoms. The energy of 1* was higher than 1 by 6.4 kcal/mol in a vacuum. DFT calculations also revealed that all Lewis acids bound to the rotated structure more strongly than to the unrotated structure, leading to the stabilization of the rotated structure. In particular, when AlCl3 is used, the rotated structure (1*/AlCl3) is more stable than the unrotated one (1/AlCl3) by 1.2 kcal/mol in a vacuum. The stabilization of the rotated structure arises from both the stronger basicity of the mu-CO ligand than the axial CO ligand and the increase of the bond strength between the mu-CO ligand and Fe(p) atom upon binding of Lewis acid to 1*. Calculation of energy barriers during electrocatalytic H2 production revealed that 1*/AlCl3 could efficiently produce H2via a chemical-electrochemical-chemical electrochemical mechanism. The analysis of the energy level of the lowest unoccupied molecular orbital showed that 1*/AlCl3 may produce H2 at significantly lower reduction potential as compared with 1*. It is also found that the catalytic activity decreases with increasing polarity of the medium. PMID- 19809729 TI - Enhanced ferromagnetic interaction in metallacyclic complexes incorporating m phenylenediamidato bridges. AB - The double stranded Cu(II)2-metallacyclic complex of formula [Cu2(mbpb)2].2H2O (1) and the triple stranded Ni(II)2-metallacyclic complexes of formula [Ni2(Hmbpb)3]PF6.21H2O (2), [Co(H2O)6][Ni2(mbpb)3)]THF.10H2O (3) and {Ag2(H2O)[Ni2(mbpb)3]}.11H2O (4) (where H2mbpb is the bisbidentate dinucleating bridging ligand 1,3-bis(pyridine-2-carboxamide) benzene) have been synthesised and characterised by single-crystal X-ray diffraction. Within the dinuclear molecules, metal ions are bridged by either fully or semideprotonated bisbidentate ligands, which are coordinated through the pyridine and amidato nitrogen donor atoms. In complex 4 the triple stranded dinuclear Ni2 units are connected to the Ag+ cations through O-amidato bridges and Ag-pi(benzene) interactions to afford a 1D bimetallic chain. Cu2 (1) and Ni2 (2-4) complexes exhibit ferromagnetic coupling between the metal ions through the bridging ligand with J(Cu-Cu) = 21.1 cm(-1) and J(Ni-Ni) in the range of 2.9-3.6 cm(-1), respectively. Amongst copper(II) dinuclear complexes bearing m-phenylenediamidato bridges, complex 1 exhibits the stronger ferromagnetic exchange coupling reported so far. DFT calculations firstly confirm that the spin polarisation mechanism is responsible for the ferromagnetic coupling, and secondly allows us to predict stronger ferromagnetic couplings in Cu(II)(2) complexes with larger tetrahedral distortions of the CuN4 coordination environment. PMID- 19809730 TI - Layered copper compounds based on 4-(3-bromothienyl)phosphonate (BTP): weak ferromagnetism observed in [Cu2(4,4'-bpy)0.5(BTP)2].H2O. AB - Two copper phosphonates based on 4-(3-bromothienyl)phosphonic acid (BTPH2) with formula Cu(H2O)(BTP) (1) and [Cu2(4,4'-bpy)0.5(BTP)2].H2O (2) are reported. Compound 1 has a layered structure in which dimers of {Cu2O2}, made up of edge sharing {CuO6} octahedra, are bridged by both the {PO3C} tetrahedra through corner-sharing and the water molecules. Compound 2 exhibits a novel layered structure where the {Cu2(BTP)2} columns, containing {Cu4O6} tetrameric cores and O-P-O linkages, are connected by 4,4'-bipyridine. In both cases, the 3 bromothienyl groups are grafted on the two sides of the layers. Magnetic studies reveal that antiferromagnetic interactions are mediated between the Cu(II) centers in 1, while for 2, weak ferromagnetism resulted from spin canting is observed. Electrochemical properties of compounds 1-2 are also investigated. PMID- 19809731 TI - Sterically hindered phosphine and phosphonium-based activators and additives for olefin polymerization. AB - The ability of phosphonium borates of the form [R3PH][B(C6F5)4], R2PHC6F4BF(C6F5)2 and R2PHC4H8OB(C6F5)3 as well as the phosphine-boranes R2PC6F4B(C6F5)2 to activate CpTiMe2(NPtBu3) for olefin polymerization was examined via both stoichiometric reactions and catalytic performance. In general these activators resulted in highly active ethylene polymerization catalysts, despite the generation of liberated phosphine donors. Independent experiments in which phosphines were added to the catalyst systems revealed the expected decrease in activity for small phosphines. However in the case of sterically encumbered phosphines, a marked increase in activity was observed. The cause of this increase is considered in the context of the concept of "frustrated Lewis pairs". PMID- 19809732 TI - Two unprecedented porous anionic frameworks: organoammonium templating effects and structural diversification. AB - Two unprecedented 3D porous anionic metal-organic frameworks, [Me2NH2]2[Cd2(bpdc)3].4dma 1 and [Me2NH2]2[Cd2(NH2bdc)3].4dma 2 (dma = N,N' dimethylacetamide, bpdc = 4,4'-biphenyldicarboxylate, NH(2)bdc = 2-amino-1,4 benzenedicarboxylate) have been solvothermally synthesized with a dimethylammonium cations template. Both 1 and 2 are constructed from low-symmetry SBUs. 1 has a chiral framework with helical nanotube-like channels, and 2 has a MOF-5-like motif. The fluorescence, N2 adsorption and ion-exchange properties for 1 have been examined. PMID- 19809733 TI - Assembly of [CunIn]-based coordination polymers from cracking the 3D framework of bulk CuI via flexible N-heterocyclic ligands. AB - Solvothermal reactions of CuI with four flexible N-heterocyclic ligands, 1,2 bis(4-pyridyl)ethane (bpe), 1,1'-(1,4-butanediyl)bis-1H-benzimidazole (bbbm), 1,1'-(1,4-butanediyl)bis-1H-benzotriazole (bbbt), and 1,1'-(1,5-pentanediyl)bis 1H-benzimidazole (pbbm), afforded four new [Cu(n)I(n)]-based coordination polymers, [Cu3I(mu-I)(mu3-I)(bpe)3]n (1), {[CuI(bbbm)1.5].0.25MeCN}n (2), and [Cu4(mu3-I)4L2]n (3: L = bbbt; 4: L = pbbm), respectively. Compounds 1-4 were characterized by elemental analysis, IR and X-ray crystallography. 1 consists of an unique 2D network in which 1D [CuI(bpe)]n zigzag chains are embroidered alternatively onto the two faces of a 2D [Cu4(mu-I)2(mu3-I)2(bpe)2]n layer through pairs of bpe bridges. 2 contains a 66-membered metallomacrocyclic [CuI(bbbm)1.5]6 unit that fuses with its neighbouring ones via sharing Cu atoms to form a 2D (6,3) layer network. 3 or 4 contains a cubanelike [Cu4(mu3-I)4] core, which works as a cross-shaped four-connecting node to link its four equivalent units to form a 2D (4,4) cross-shaped network () or acts as a Y-shaped three-connecting node to connect its three equivalent ones to form a 1D double chain (). In addition, the photoluminescent properties of in solid state at ambient temperature were also investigated. PMID- 19809734 TI - Optically active uniform potassium and lithium rare earth fluoride nanocrystals derived from metal trifluroacetate precursors. AB - This paper reports the first systematical synthesis of near-monodisperse potassium and lithium rare earth (RE) fluoride (K(Li)REF4) nanocrystals with diverse shapes (cubic KLaF4 and KCeF4 wormlike nanowires, nanocubes and nanopolyhedra; cubic LiREF4 (RE = Pr to Gd, Y) nanopolyhedra; tetragonal LiREF4 (RE = Tb to Lu, Y) rhombic nanoplates) via co-thermolysis of Li(CF3COO) or K(CF3COO) and RE(CF3COO)3 in a hot oleic acid/oleylamine/1-octadecene solution. The effects of the solvent composition, reaction temperature and time on the crystal phase purity, shape, and size of the as-prepared nanocrystals have been investigated in detail. The formation of monodisperse nanocrystals is found to strongly depend upon the nature of both alkali metals from Li to K, and the rare earth series from La to Lu and Y. Based on the series of experimental results, a controlled-growth mechanism has also been proposed. In addition, the ease of doping of these as-synthesized host nanocrystals for designed luminescence properties is assessed. For example, monodisperse and single-crystalline Eu3+ doped KGdF4, Yb3+ and Er3+ co-doped LiYF4 nanocrystals redispersed in cyclohexane exhibit visible room-temperature red and green emissions under ultraviolet (UV) excitation and near infrared (NIR) 980 nm laser excitation, respectively. PMID- 19809735 TI - Palladium and platinum complexes of tellurium-containing imidodiphosphinate ligands: nucleophilic attack of Li[(P(i)Pr2)(TeP(i)Pr2)N] on coordinated 1,5 cyclooctadiene. AB - Homoleptic group 10 complexes of ditellurido PNP (PNP = imidodiphosphinate), heterodichalcogenido PNP and monotellurido PNP ligands, M[(TeP(i)Pr2)2N]2 (1: M = Pd; 2: M = Pt), M[(EP(i)Pr2)(TeP(i)Pr2)N]2 (3: M = Pd, E = Se; 4: M = Pt, E = Se; 5: M = Pd, E = S; 6: M = Pt, E = S) and M[(P(i)Pr2)(TeP(i)Pr2)N]2 (7: M = Pd; 8: M = Pt), respectively, were prepared by metathesis between alkali-metal derivatives of the appropriate ligand and MCl2(COD) in THF. Complexes 1-8 were characterised in solution by multinuclear (31P, 77Se, 125Te and 195Pt) NMR spectroscopy and, in the case of 1, 2, trans-7, cis-7 and trans-8, in the solid state by X-ray crystallography. The square-planar complexes 3-6 are formed as a mixture of cis- and trans-isomers on the basis of NMR data. The cis and trans isomers of 7 were separated by crystallisation from different solvents. In addition to trans-8, the reaction of Li[(P(i)Pr2)(TeP(i)Pr2)N] with MCl2(COD) produced the heteroleptic complex Pt[(P(i)Pr2)(TeP(i)Pr2)N][sigma:eta2 C8H12(P(i)Pr2NP(i)Pr2Te)] (9) resulting from nucleophilic attack on coordinated 1,5-cyclooctadiene. Complex 9 was identified by multinuclear (13C, 31P, 125Te and 195Pt) NMR spectroscopy, which revealed a mixture of geometric isomers, and by X ray crystallography. PMID- 19809736 TI - Photoresponsive SAMs on gold fabricated from azobenzene-functionalised asparagusic acid derivatives. AB - We have prepared a range of azobenzene derivatives equipped with an asparagusic acid-based 1,2-dithiolane headgroup suitable for chemisorption on solid gold substrates. The formation of self-assembled monolayers (SAMs) of the amide cyclo S2C3H5-4-C(O)NH-p-C6H4-N=N-Ph (1) and the ester cyclo-S2C3H5-4-C(O)O-p-C6H4-N=N Ph (2) on gold was monitored in situ and in real time by optical second harmonic generation (SHG). The structure and composition of these SAMs was investigated by a range of ex situ methods, viz. ellipsometry, X-ray photoelectron spectroscopy (XPS), near-edge X-ray absorption fine structure (NEXAFS) spectroscopy and Fourier transform infrared reflection absorption spectroscopy (FTIRRAS). Reversible, but moderate, photoswitchability was observed for these one-component SAMs by ellipsometry and dynamic contact angle measurements. Use of a second 1,2 dithiolane component for lateral dilution of the photoactive terminal groups resulted in a much more pronounced photoresponse. PMID- 19809737 TI - Thiacrown Pt(II) complexes with group 15 donor ligands: pentacoordination in Pt(II) complexes. AB - We report the synthesis and full characterization for a series of thiacrown complexes of Pt(II) incorporating the fluxional trithiacrown ligand 1,4,7 trithiacyclononane ([9]aneS3) and several group 15 donors ligands. Reaction of [Pt([9]aneS3)Cl2] with a full stoichiometric equivalent of the group 15 donor (L = 2 x AsPh3, SbPh3 or 1,2-bis(diphenylarsenio) ethane (dpae) followed by metathesis with NH4PF6 yields [Pt([9]aneS3)L](PF6)2. We also report the analogous Pd(II) complex with dpae. Similar reactions of the starting Pt complex with one equivalent of XPh3 (X = As or Sb) result in complexes of the formula [Pt([9]aneS3)(XPh3)(Cl)](PF6). All six new complexes have been fully characterized by multinuclear NMR, IR, and UV-Vis spectroscopies in addition to elemental analysis and single crystal structural determinations. The X-ray structures of each complex indicate an axial M-S interaction formed by the endodentate conformation of the [9]aneS3 ligand. The axial M-S distance is highly dependent upon the ancillary donor set. The axial M-S distance shortens with the identity of the group 15 donor ligand according to the trend, Sb < As < P, due to their increasingly poorer donor qualities. The two bis pnictogen complexes, [Pt([9]aneS3)(AsPh3)2](PF6)2 and [Pt([9]aneS3)(SbPh3)2](PF6)2 form unusual five coordinate distorted trigonal bipyramids in contrast to the pseudo-five coordinate, elongated square pyramidal structures typically observed in Pt(II) complexes of [9]aneS3. The distortion arises from intramolecular pi-pi interactions between the phenyl rings on the two different triphenyl ligands. Chemical shifts in the 195Pt NMR also show similar periodic relationships which trend progressively upfield as the donor atom becomes larger. As expected, the coordinated [9]aneS3 ligand shows fluxional behavior in its NMR spectra, resulting in a single 13C NMR resonance, despite the asymmetric coordination environment found in both chloro complexes. The line width for the carbon NMR resonance as well as for the 195Pt NMR peak is highly sensitive to the nature of the group 15 donor, with poorer donors such as SbPh3 showing significant line broadening. Measurements from the electronic spectra support that the ligand field strength of the pnictogen donor decreases with its increasing size. PMID- 19809738 TI - Iron(III) citrate speciation in aqueous solution. AB - Citrate is an iron chelator and it has been shown to be the major iron ligand in the xylem sap of plants. Furthermore, citrate has been demonstrated to be an important ligand for the non-transferrin bound iron (NTBI) pool occurring in the plasma of individuals suffering from iron-overload. However, ferric citrate chemistry is complicated and a definitive description of its aqueous speciation at neutral pH remains elusive. X-Ray crystallography data indicates that the alcohol function of citrate (Cit4-) is involved in Fe(III) coordination and that deprotonation of this functional group occurs upon complex formation. The inability to include this deprotonation in the affinity constant calculations has been a major source of divergence between various reports of iron(III)-citrate affinity constants. However the recent determination of the alcoholic pKa of citric acid (H4Cit) renders the reassessment of the ferric citrate system possible. The aqueous speciation of ferric citrate has been investigated by mass spectrometry and EPR spectroscopy. It was observed that the most relevant species are a monoiron dicitrate species and dinuclear and trinuclear oligomeric complexes, the relative concentration of which depends on the solution pH value and the iron : citric acid molar ratio. Spectrophotometric titration was utilized for affinity constant determination and the formation constant for the biologically relevant [Fe(Cit)2]5- is reported for the first time. PMID- 19809739 TI - Synthesis and characterisation of PCsp3P phosphine and phosphinite iridium complexes. Cyclometallation and dehydrogenation of a cyclohexyl ring. AB - A cyclohexyl based pincer phosphine ligand undergoes cyclometalation with an iridium cyclooctadiene precursor to give a highly thermally stable iridium(III) complex where a C(sp3)-H bond has been oxidatively added. This iridium(III) hydride complex is reduced with potassium to give a terminal iridium(I) dinitrogen complex with no tendency to dimerization. The corresponding cyclohexyl phosphinite ligand undergoes reversible dehydrogenation to give the aromatic cyclometalated iridium(III) hydride complex together with 3 equivalents of dihydrogen. PMID- 19809740 TI - Deprotonation and reductive addition reactions of hypervalent aluminium dihydride compounds containing substituted pyrrolyl ligands with phenols, ketones, and aldehydes. AB - The reactivities of [C4H2N(CH2NMe2)2]AlH2 (1) with primary and secondary amines, phenols, ketones, and phenyl isothiocyanate were examined. Reactions of 1 with one or two equivalents of 2,6-dichloroaniline in methylene chloride generated [C4H2N(CH2NMe2)2]AlH(NHC6H3-2,6-Cl2) (2) and [C4H2N(CH2NMe2)2]Al(NHC6H3-2,6-Cl2)2 (3), respectively, following hydrogen elimination. Similarly, the reactions of 1 with one or two equivalents of carbazole afforded [C4H2N(CH2NMe2)2]AlH(NC12H8) (4) or [C4H2N(CH2NMe2)2]Al(NC12H8)2 (5) by deprotonating the acidic N-H of carbazole. Reacting 1 with one equivalent of 2,6-diisopropylphenol in diethyl ether formed an aluminium phenoxo compound [C4H2N(CH2NMe2)2]AlH(OC6H3-2,6-iPr2) (6), by deprotonation of phenol as well with the elimination of one equivalent hydrogen. Further reaction of 6 with one equivalent of 2,4,6 trimethylacetophenone in methylene chloride generated [C4H2N(CH2NMe2)2]Al(OC6H3 2,6-iPr2)[OC(=CH2)(C6H2-2,4,6-Me3)] (7) by deprotonating the methyl proton of the acetophenone. Similar deprotonation occurred when 1 reacted with two equivalents of 2,4,6-trimethylacetophenone in methylene chloride to generate [C4H2N(CH2NMe2)2]Al[OC(=CH2)(C6H2-2,4,6-Me3)]2 (8). Compounds [C4H2N(CH2NMe2)2]Al(OCHPh2)2 (9), and [C4H2N(CH2NMe2)2]Al(SCHNPh)2 (10) could also be obtained by reacting 1 with two equivalents of benzophenone and phenyl isothiocyanate, respectively through hydroalumination. The 1H NMR spectra of 10 showed broad signals for the CH2N and NMe2 groups, which represent dynamical fluctuations of the molecules in solution state. The estimated energy barrier (DeltaG(c)(double dagger)) from the coalescence temperature for the fluctuation was estimated at 17.1 Kcal mol(-1). The solid-state structures of compounds 2, 3, 5, 7, 9, and 10 have been determined. PMID- 19809741 TI - Electrochemical synthesis and structural characterization of Co(II), Ni(II) and Cu(II) complexes of N,N-bis(4,5-dimethyl-2-hydroxybenzyl)-N-(2 pyridylmethyl)amine. AB - The electrochemical oxidation of anodic metal (cobalt, nickel or copper) in a cell containing an acetonitrile solution of the ligand N,N-bis(4,5-dimethyl-2 hydroxybenzyl)-N-(2-pyridylmethyl)amine (H2L) affords complexes [Co2L2].H2O (1), [Ni3L3] (2) and [Cu2L2] 3H2O (4). On using nickel as the anode and the addition to the solution electrolytic phase of the amount of water necessary to saturate the solution, the electrolytic process gave rise to the new compound [Ni2L2(H2O)1.5].CH3CN (3). Compounds 1 and 4 are dimeric and the metal atoms are pentacoordinated. Compound 3 also consists of dimeric neutral molecules with the nickel atoms in both penta- and hexacoordinated environments. The crystal structure of shows the presence of a trimeric compound in which the nickel atoms are hexacoordinated. Electronic, IR and FAB spectra of the complexes are discussed and related to the structural information. The magnetic behavior of 1-4 denotes the occurrence of intramolecular antiferromagnetic interactions. The values obtained for the coupling constant J are -4.2 cm(-1), -5.3 cm(-1), -30 cm( 1) and -4.7 cm(-1) for 1, 2, 3 and 4, respectively. These values are in full agreement with the structural characteristics of the compounds. The catalytic activity of the complexes towards the decomposition of hydrogen peroxide (catalase activity) was also studied. PMID- 19809742 TI - 5,6-Membered palladium pincer complexes of 1-thiophosphoryloxy-3 thiophosphorylbenzenes. Synthesis, X-ray structure, and catalytic activity. AB - Novel unsymmetrical ligands, 1-thiophosphoryloxy-3-thiophosphorylbenzenes 3a-d, bearing phosphine sulfide and thiophosphoryloxy moieties as coordinating sites, were found to undergo cyclometalation at the C-2 position of the central benzene ring in a reaction with bis(benzonitrile)palladium dichloride affording rare examples of nonsymmetrical pincer complexes, namely [2-{(thiophosphoryl)oxy}-6 (diphenylthiophosphoryl)phenyl]palladium chlorides 4a-d, containing 5- and 6 membered fused metallacycles with kappa3-SCS'-coordination. Molecular structures of the complexes were characterized by X-ray diffraction. These complexes demonstrated high catalytic activity for the Suzuki cross-coupling reactions of aryl bromides with phenylboronic acid. PMID- 19809743 TI - Synthesis and structures of complexes with axially chiral isoquinolinyl naphtholate ligands. AB - The synthesis of axially chiral ligands 1-(3',6'-di-t-butyl-2'-hydroxy-1' naphthyl)-3-R-isoquinoline (R = H, Pr(i), Bu(t)) (LR-H) is described. Ligands with unsubstituted isoquinolinyl rings tend to give 1:2 metal complexes. The syntheses and crystal structures of Li2(LH)2(THF)2 (9), (LH)2Ti(OPr(i))2 (12), Zn(LH)2 (13) and [Mg(LH)2]2 (14) are reported. Complex formation is highly stereoselective; the ligands in 1:2 complexes have the same stereochemistry (i.e. R,R and S,S but not R,S), whereas in the binuclear magnesium compound 14 the bridging and non-bridging ligands LH have opposite stereochemistry. The reaction of LH-H with Pd(acac)2 afforded the N,O chelate Pd(acac)(LH) (10), whereas towards K2PtCl4 the same ligand acts as an N-donor only, to give trans-PtCl2(LH H)2 (11) in which the OH groups are hydrogen-bonded to one of the two chloride ligands. The more bulky ligand with a t-butyl substituent in the 3-position of the isoquinolinyl ring reacts with zinc and magnesium bis(amides) to give the mixed-ligand species (LBu)ZnN(SiMe3)2 and (LBu)MgN(SiMe3)2, respectively, which catalyse the ring-opening polymerisation of epsilon-caprolactone (CL) and rac lactide (LA). PMID- 19809744 TI - Synthesis, characterization, electrochemistry and ion-binding studies of ruthenium(II) bipyridine receptor molecules containing calix[4]arene-azacrown as ionophore. AB - A number of molecular receptors containing ruthenium(II) bipyridine moiety as fluorophore and calix[4]arene-azacrown hybrid molecule as ionophore have been synthesized and characterized. These receptors (1-4) exhibit strong 3MLCT luminescence bands in the range 613-618 nm. The cation-binding property of these fluoroionophores have been investigated with the ions Na+, K+, Mg2+, Ca2+, Cs+, Zn2+, Cd2+, Hg2+ and Pb2+ and the recognition event monitored by luminescence, 1H NMR spectroscopy, the oxidation potential of metal ion and UV/Vis absorption studies. The luminescence study suggests complexation of all four receptors with Zn2+, Cd2+ and Hg2+ and for 2 and 3 also with Pb2+. The binding constants (Ks, except 4) and stoichiometries of the complexes have been calculated from the luminescence titration data, with values of Ks ranging from 2.53 x 10(5) to 6.27 x 10(3) M(-1). The 1H NMR spectroscopy study exhibits interactions of Na+ and K+ with 1 and 4, K+ and Cs+ with 2, and K+ with 3. Binding constants with these metal ions have been calculated from 1H NMR titrations using computer programs or by a direct method depending on the type of change in the chemical shifts observed upon addition of metal ions. Stoichiometry of the complexes has been determined from Job's plot. Electrochemical properties of 1-4 have been studied in the absence and also in the presence of selected metal ions and the shift of the oxidation potentials of the Ru(II) in the presence of guest ions suggests an interaction of Na+, Hg2+ and Pb2+ with 1 and 2 but not with 3 and 4, indicating a selective electrochemical response towards certain metal ions. The results obtained are presented and discussed in light of ion-binding properties and methods of detection. PMID- 19809745 TI - Low-valence oxo-centred triruthenium complexes by bridging acetate substitution with pyrazolyldiazine or pyridinyltetrazine ligands. AB - A series of oxo-centred triruthenium-acetate complexes with valence III,III,II were prepared by reactions of [Ru3(III,III,III)]+ precursor [Ru3O(OAc)6(py)2(CH3OH)]+ (1) with 3-chloro-6-(pyrazol-1-yl)pyridazine (cppd), 3 chloro-6-(3,5-dimethylpyrazol-1-yl)pyridazine (cmppd), 3,6-bis(pyrazol-1 yl)pyridazine (ppd), 3,6-bis(3,5-dimethylpyrazol-1-yl)pyridazine (mppd) or 3,6 bis(2-pyridyl)-1,2,4,5-tetrazine (bptz). When neutral Ru3(III,III,II) precursor Ru3O(OAc)6(CO)(CH3OH)2 (2) was utilized, a stable low-valence Ru3(III,II,II) derivative complex was successfully isolated. As established through crystal structural analyses, they were derived from 1 or 2 by substitution of the axial methanol and one of six bridging acetates in the parent Ru3(mu3-O)(mu-OAc)6 cluster core as well as one axial pyridine in some cases. As revealed by electrochemical and spectroscopic studies, substituting one of the six bridging acetates in the parent Ru3(mu3-O)(mu-OAc)6 cluster core significantly modifies the electronic and redox characteristics. Compared with those for the parent compound [Ru3O(OAc)6(py)3]+, triruthenium-based redox potentials of these derivatives show remarkable positive shifts. PMID- 19809746 TI - Water-stable ammonium-terminated carbosilane dendrimers as efficient antibacterial agents. AB - A new family of amine- and ammonium-terminated carbosilane dendrimers of the type Gn-[Si(CH2)3N(Et)CH2CH2NMe2]x and Gn-{[Si(CH2)(3)N+R(Et)CH2CH2N+RMe2]x(X-)y} (where n = 1, 2 and 3; R = H, X = Cl; R = Me, X = I) respectively has been synthesized by hydrosilylation of N,N-dimethyl-N'-allyl-N'-ethyl-ethylenediamine, [(CH2=CH-CH2)(Et)N(CH2)2NMe2] with the corresponding hydride-terminated dendrimers and subsequent quaternization with HCl or MeI. Quaternized dendrimers are soluble and stable in water or other protic solvents for long time periods. The antibacterial properties of the quaternary ammonium functionalized dendrimers have been evaluated showing that they act as potent biocides in which the multivalency along with the biopermeability of the carbosilane dendritic skeleton play an important role in the antibactericidal activity of these compounds. PMID- 19809747 TI - Coordination modes of bridge carboxylates in dinuclear manganese compounds determine their catalase-like activities. AB - To explore the role of bridge carboxylate coordination modes on the catalase-like activities of dinuclear manganese compounds, [Mn(II)2(bpmapa)2(H2O)2](ClO4)2 (1), [Mn(II)2(pbpmapa)2(H2O)2](ClO4)2 (2), and [Mn(II)2(bpmaa)2(H2O)3](ClO4)2 (3) (bpmapa = [bis(2-pyridylmethyl)amino]propionic acid, pbpmapa = alpha-phenyl-beta [bis(2-pyridylmethyl)amino]propionic acid, and bpmaa = [bis(2 pyridylmethyl)amino]acetic acid), in which Mn(II)-Mn(II) centers have a similar coordination sphere but different carboxylate-Mn bridging modes have been synthesized and structurally characterized by single X-ray diffraction, UV visible, IR, and EPR spectroscopies, and their catalase-like activities were investigated. Studies of their catalytic activities and the influence of the nitrogenous bases on their catalytic activities indicated that the carboxylate-Mn coordination mode was crucial in H2O2 deprotonation, and eventually in H2O2 disproportionation. Compound 1 with a bidentate carboxylate bridge showed higher catalase-like activity than 2 and 3, in which the carboxylate groups have a monodentate bridging mode. The deprotonation ability of the carboxylate anion was determined by the O-C-O angle and the distance between the weakly bound oxygen of the bridging carboxylate to the manganese ion. The smaller the angle, and the shorter the distance, the stronger the basicity that the carboxylate anion exhibits. The bidentate mu-1,1 bridging coordination mode functionally mimicked the glutamate residues at the manganese catalase active site. Our results suggested that increasing the basicity of the bridging carboxylate ligand of the catalase model compounds will increase their deprotonation ability and lead to more active catalase mimics. PMID- 19809748 TI - A novel route to rhodaboratranes [Rh(CO)(PR3){B(taz)3}]+ via the redox activation of scorpionate complexes [RhLL'Tt]. AB - The reaction of a mixture of the sodium salts of dihydrobis(4-ethyl-3-methyl-5 thioxo-1,2,4-triazolyl)borate, NaBt, and hydrotris(4-ethyl-3-methyl-5-thioxo 1,2,4-triazolyl)borate, NaTt, with [{Rh(cod)(mu-Cl)}2] gave [Rh(cod)Bt] and [Rh(cod)Tt], which separately react with CO gas to give the unstable dicarbonyl [Rh(CO)2Bt] and an equilibrium mixture of two isomers of [Rh(CO)2Tt] and [(RhTt)2(mu-CO)3], respectively. Tertiary phosphorus donor ligands react with the mixture of [Rh(CO)2Tt] and [(RhTt)2(mu-CO)3] to give [Rh(CO)(PR3)Tt] (R = Cy, NMe(2), Ph or OPh) and [Rh{P(OPh)3}2Tt] in which rhodium is bound to two sulfur atoms of the scorpionate ligand; the B-H bond is directed towards the metal to give an agostic-like B-H...Rh interaction. Dinuclear [(RhTt)2(mu-CO)3] has kappa3[S3]-bound Tt ligands with a rhodium-rhodium bond bridged by three carbonyls. In solution the mononuclear Tt complexes undergo rapid dynamic interchange of the three thioxotriazolyl rings, probably via kappa3[S3] coordinated intermediates. The monocarbonyls [Rh(CO)(PR3)Tt] (R = Cy, NMe2 or Ph) react with two equivalents of [Fe(eta-C5H5)2][PF6] in the presence of triethylamine to give the monocationic rhodaboratranes [Rh(CO)(PR3){B(taz)3}]+, with boron NMR spectroscopy providing evidence for the boron-rhodium bond. In the solid state, rhodium is bound to the three sulfur atoms and the boron of the B(taz)3 fragment, forming a tricyclo[3.3.3.0] cage. The phosphine is trans to the Rh-B bond, the long Rh-P bond indicating a pronounced trans influence for the coordinated boron. The cation [Rh(CO)(PPh3){B(taz)3}]+ reacts with [NBu(n)(4)]I to give [Rh(PPh3)I{B(taz)3}], in which the halide is trans to the Rh-B bond, and a second species, possibly [Rh(CO)I{B(taz)3}]. The dirhodaboratrane [Rh2(PCy3){B(taz)3}2][PF6]2, a minor byproduct in the synthesis of [Rh(CO)(PCy3){B(taz)3}][PF6], has a distorted square pyramidal rhodium atom with a vacant site trans to the Rh-B bond. The second metal has four coordination sites filled by the sulfur and boron atoms of a second B(taz)3 unit, the remaining octahedral sites occupied by two of the sulfur atoms of the first B(taz)3 unit which therefore bridges the two rhodium atoms. PMID- 19809749 TI - Neutron total scattering study of the delta and beta phases of Bi2O3. AB - The highly disordered structure of the delta phase of Bi2O3, which possesses the highest known oxide-ion conductivity, has been studied using neutron powder diffraction. A detailed analysis of data collected at 1033(3) K using Rietveld refinement indicates that the time-averaged structure of delta-Bi2O3 can be described using the accepted model of a disordered, anion-deficient fluorite structure in space group Fm3m. However, reverse Monte Carlo modelling of the total (Bragg plus diffuse) scattering demonstrates that the local anion environment around the Bi3+ resembles the distorted square pyramidal arrangement found within the stable alpha and metastable beta phases at ambient temperature, which is characteristic of the cation's 6s2 lone-pair configuration. Similarities between the structures of the highly disordered delta phase and the ambient temperature metastable beta phase are used to support this assignment and assess the validity of previous structural models based on short-range ordering of vacancies within the cubic lattice of delta-Bi2O3. PMID- 19809750 TI - Studies on bis(halogeno) dioxomolybdenum(VI)-bipyridine complexes: synthesis and catalytic activity. AB - Dioxomolybdenum(VI) complexes with the general formula [MoO2Cl2L2] (L2=3,3' dimethyl-2,2'-bipyridine, 5,5'-dimethyl-2,2'-bipyridine, 6,6'-dimethyl-2,2' bipyridine, 4,4'-dibromo-2,2'-bipyridine, 5,5'-dibromo-2,2'-bipyridine, 5,5' diamino-2,2'-bipyridine; 5,5'-dinitro-2,2'-bipyridine; 5,5'-di-ethoxycarbonyl 2,2'-bipyridine; 6-phenyl-2,2'-bipyridine; 2,2':6',2''-terpyridine) have been prepared and characterised. [MoO2Cl2(5,5'-di-ethoxycarbonyl-2,2'-bipyridine)] has been examined by single crystal X-ray analysis. The complexes were applied as homogenous catalysts for the epoxidation of cyclooctene with tert-butyl hydroperoxide (TBHP) as oxidising agent. The new compounds show an overall high activity and are highly selective catalysts in the epoxidation of cyclooctene. The stability of the complexes and differences in the catalytic activity can be clearly attributed to electronic contributions of the functional groups on bipyridine ligands and to steric restrictions. DFT calculations have assisted in a better understanding of the stability of the complexes and are in agreement with experiment. The influence of the terminal oxo ligands and the Lewis base ligands on the Mo center keep the compounds on quite a stable level of electron density. PMID- 19809751 TI - Mono- and dinuclear manganese(III) complexes showing efficient catechol oxidase activity: syntheses, characterization and spectroscopic studies. AB - Four side-off compartmental ligands L1-L4 [L1 = N,N'-ethylenebis(3-formyl-5 methyl-salicylaldimine), L2 = N,N'-1-methylethylenebis(3-formyl-5 methylsalicylaldimine), L3 = N,N'-1,1-dimethylethylenebis(3-formyl-5 methylsalicylaldimine) and L4= N,N'-cyclohexenebis(3-formyl-5 methylsalicylaldimine)] having two binding sites, N2O2 and O4, have been chosen to synthesize mononuclear and dinuclear manganese(III) complexes with the aim to study their catecholase activity using 3,5-di-tert-butylcatechol (3,5-DTBC) as substrate in the presence of molecular oxygen. In all cases only mononuclear manganese complexes (1-4) were obtained, with manganese coordination taking place at the N2O2 binding site only, irrespective of the amount of manganese salt used. All these complexes have been characterized by routine physico-chemical techniques. Complex MnL2Cl.4H2O (2) has further been structurally characterized by X-ray single crystal structure analysis. Four dinuclear manganese complexes, 5 8, were obtained after condensing the two pending formyl groups on each ligand (L1-L4) with aniline followed by reaction with MnCl2 to put the second Mn atom onto another N2O2 site. The catalytic activity of all complexes 1-8 has been investigated following the oxidation of 3,5-di-tert-butylcatechol (3,5-DTBC) to 3,5-di-tert-butylbenzoquinone (3,5-DTBQ) with molecular oxygen in two different solvents, methanol and acetonitrile. The study reveals that the catalytic activity is influenced by the solvent and to a significant extent by the backbone of the diamine and the behavior seems to be related mainly to steric rather than electronic factors. Experimental data suggest that a correlation, the lower the E(1/2) value the higher the catalytic activity, can be drawn between E(1/2) and Vmax of the complexes in a particular solvent. The EPR measurements suggest that the catalytic property of the complexes is related to the metal center(s) participation rather than to a radical mechanism. PMID- 19809755 TI - [The WHO and its "World Alliance for Patient Safety" iniciative]. PMID- 19809756 TI - [Dengue 2009: chronology of an epidemic]. PMID- 19809757 TI - [Respiratory tract infections and wheezing in children. What role does rhinovirus play?]. PMID- 19809758 TI - [Prenatal diagnosis by the presence of fetal DNA in maternal blood]. PMID- 19809759 TI - [Nutrition status in Argentinean children 6 to 72 months old: results from the National Nutrition and Health Survey (ENNyS)]. AB - INTRODUCTION: Data availability on nutrition status from vulnerable population groups is essential for the design and evaluation of interventions. The Ministry of Health from Argentina developed in 2004-2005 the National Survey of Nutrition and Health. Our objective is to describe the nutrition status of infants and preschool children (6-72 months), globally and by region and socioeconomic level. POPULATION AND METHODS: A probabilistic, multistage sample was selected, representative at the national, regional and provincial levels. Anthropometric indices weight/age, height/age and weight/height, were estimated, according to WHO reference. Nutrient intake was estimated by a 24-hours recall, and hemoglobin, serum ferritin, retinol and vitamin D concentration were measured. RESULTS: Prevalence of stunting, wasting and obesity were 8.0%, 1.3%, and 10.4%, respectively. Prevalence of anemia was 16.5% in children <6 years and 35.3% in children aged 6-23 months. Prevalence of subclinical vitamin A deficiency in children of 2-5 years was 14.3%, and prevalence of vitamin D deficiency in Patagonia was 2.8% in children aged 6-23 months. Different nutrient intake inadequacies were observed. Infants and children at the lower socioeconomic level showed higher prevalence of nutrition inadequacies. CONCLUSIONS: The coexistence of different nutrition deficiencies and overweight and obesity constitute the main characteristic of Argentinean infants and preschool children. Relevant differences exist according to socio-economic and geographic conditions. PMID- 19809760 TI - [Non invasive prenatal genetic diagnosis of fetal RhD and sex through the analysis of free fetal DNA in maternal plasma]. AB - INTRODUCTION: The analysis of free fetal DNA in maternal plasma allows the assessment of fetal genetic material avoiding the necessity of invasive procedures during pregnancy. OBJECTIVE: To evaluate the feasibility and the diagnostic performance of fetal sex and fetal RhD detection through the analysis of free fetal DNA in maternal plasma using standard reagents in molecular biology. MATERIAL AND METHODS: A hundred and nine blood samples of pregnant women were obtained. Amplification by real time PCR a sequence from the RhD gene in Rh negative patients and a Y-chromosome sequence, for the diagnosis of fetal Rh and sex respectively, were performed. Results were compared with neonatal outcomes. RESULTS: From the 109 samples, 26 are still ongoing, 4 ended in spontaneous abortions and in 3 were lost to follow up. From the remaining 76 samples with neonatal result, the determination of fetal Rh from the RhD gene was performed in 65 Rh negative women, whereas in 66 samples the fetal sex analysis was evaluated. Overall, 15 samples had not conclusive results and were excluded from the study. The predictive values for RhD positive and negative were 85% and 90%, respectively, while the prediction for male sex was 94.3% and for female sex 95%. CONCLUSION: The non invasive determination of fetal RhD and sex in maternal plasma using standard reagents in molecular biology was feasible in the majority of the samples, with a diagnostic performance similar to the reported in the literature. PMID- 19809761 TI - [Growth hormone treatment in younger than six years of age short children born small for gestational age]. AB - INTRODUCTION: Approximately 10% of children born small for gestational age (SGA) do not show spontaneous catch-up growth. OBJECTIVE: Our objective was to assess the efficacy and safety of 2 years growth hormone treatment in children younger than 6 years of age, born SGA. MATERIAL AND METHODS: Fourteen patients, mean age 4.2 + or - 1.1 years were treated with growth hormone at 1.0 UI/kg/wk for two years. RESULTS: Growth velocity increased from a mean + or - SD of 5.4 + or - 1.7 cm/yr to 9.8 + or - 1.50 cm/yr and 7.6 + or - 1.5 cm/yr during the first year and second year, respectively (p 0.00058). Serum IGF-I and its binding protein BP3 increased significantly throughout treatment. Basal glucose and insulin levels increased significantly during treatment, p= 0.0006 and p= 0.036, respectively, without significant changes in postprandial glucose or insulin levels. A mild change in insulin sensitivity was observed along treatment. CONCLUSION: Two years growth hormone treatment induced a significant growth acceleration in children born small for gestational age, allowing them to attain a normal height. No serious adverse events were reported. PMID- 19809762 TI - [Cardiac surgical repair in newborns: five years' experience in neonatal open surgery]. AB - INTRODUCTION: Four hundred newborns die every year in our country suffering from congenital heart disease. Definitive surgical repair, whenever possible, is nowadays the optimal therapeutic strategy. Our goal is to describe mortality and morbidity in neonatal surgery with cardiopulmonary bypass in a tertiary public hospital in Argentina. PATIENTS AND METHODS: Descriptive, retrospective study. Every patient, younger than 45 days, with cardiac surgery requiring cardiopulmonary bypass, at the Garrahan Hospital between 2004 and 2008 was included. Demographic, surgical and postoperative data were collected. Adjusted mortality risk analysis, and descriptive statistics from the most frequent diagnosis were performed. Results are expressed as median and rank or percentage. RESULTS: 200 newborns were operated, 62% males. Median age was 21 days (r 1- 45) and median weight 3.1 kg (r 1.6-6.2). Total anomalous pulmonary venous return, transposition of great arteries an hypoplastic left heart syndrome diagnoses accounted for 75% of the procedures. Median length of stay was 12 days (r 0-191), and 6 days of mechanical ventilation (r 0-180). Eighteen percent of the patients required peritoneal dialysis. Whole series mortality was 19% and fell to 14% in 2008. Unstable preoperative condition and postoperative complications increased mortality, OR= 2.23 (1.02-4.89) and OR= 10 (3.6-33.4), respectively. CONCLUSIONS: Our postoperative mortality is similar to those reported in foreign countries databases. Patients with unstable preoperative condition and post-operative complications had higher mortality. PMID- 19809763 TI - [A non invasive method for assessing sexual development at adolescence]. AB - INTRODUCTION: Observational assessments of puberty that invades the adolescent's privacy are not acceptable for research in population groups. Results based on self assessment have been variable, and in many cases poor. OBJECTIVES: To evaluate the validity of a questionnaire with simple questions addressed to assess early, intermediate and advanced puberty periods rather than specific stages. MATERIAL AND METHODS: In an outpatient clinic at the Service of Adolescence of a public hospital, 188 and 142 healthy girls and boys aged 8-18 years, and 36 girls and boys aged 8-9 years attending a public school were studied. Adolescents attended the Service for the first time. Those with chronic diseases were excluded from the study. Children answered the questionnaire before entering the doctor's office, where a trained professional clinically assessed their puberal development (Tanner's stage). The questionnaire was previously tested in 30 adolescents. RESULTS: The highest concordance were found in the questions: "Have you started puberty?", with Tanner's stages III, IV or V (Kappa value= 0.60); "Have you already had your first menstrual period?" with stages IV V (K= 0.69); and "Do you shave?" with stages IV-V (K= 0.66). In most cases, these questions showed high (> or = 0.80) sensitivity and specificity for detecting the mentioned puberty periods. CONCLUSIONS: The method showed to be reliable, and its further evaluation in a non medical setting (schools, households, etc.) is recommended. PMID- 19809764 TI - [Comparing the clinical-functional state in children with cystic fibrosis detected by neonatal screening or by clinical symptoms]. AB - INTRODUCTION: Newborn Screening for Cystic Fibrosis (CF) has demonstrated better clinical and functional results in patients diagnosed and treated prematurely. OBJECTIVE: To compare clinical and functional state in children with CF detected by newborn screening or by symptoms. PATIENTS AND METHODS: Cross-sectional study. We compared two groups paired by age, sex and genotype. A group detected by newborn screening (Group N) and other by symptoms (Group S), both confirmed with sweat test. We evaluated: age at diagnosis and enter to the center, z score weight/age (z W/A) at first visit and z score for body mass index (BMI) and height/age (z H/A) in last control, score of Shwachman-Brasfield (S/B), forced expiratory volume in one second (FEV1), first isolation of Pseudomonas aeruginosa (Pa) and number of hospitalizations. RESULTS: We included 21 patients in each of the groups from an universe of 250 patients treated in our center. The mean of both groups was 4.4 years. The age at diagnosis was lower in the group N than in the group S (p= 0.002, IC95%:-1.11/- 0.32). z W/A at diagnosis; z IMC; z H/A, score S/B and FEV(1) were better in group N (p< 0.001, 0.3, 0.01, 0.02, 0.1, respectively). The average age of first isolation of Pa and the average of hospitalizations was lower in group N (p= 0.34, IC95%: -1.45/0.51, p= 0.04, IC95%: -3.16/-0.07). CONCLUSIONS: Children with CF detected by neonatal screening programs had better clinical and functional outcome. PMID- 19809765 TI - [Renal manifestations of tuberous sclerosis in children]. AB - INTRODUCTION: Tuberous sclerosis is a neurocutaneous disorder characterized by the growth of hamartomas in multiple organs. Renal involvement is frequent and is the second cause of morbidity and mortality in these patients. OBJECTIVES: To describe de prevalence, type and long-term outcome of renal involvement in children with tuberous sclerosis. PATIENTS AND METHODS: We retrospectively reviewed clinical notes and renal ultrasounds of 38 children followed between 1989 and 2008 who met diagnostic criteria for tuberous sclerosis. RESULTS: Median patient age was 10 years and median follow-up was 6.7 years; 44.7% (17 patients, 11 females) showed renal manifestations diagnosed by ultrasound screening in 13, in 2 by palpable mass and in another 2 by hypertension. On initial evaluation, renal ultrasound was abnormal in 7 children (18.4%); during followup another 10 patients (26.3%) developed renal lesions. Renal involvement included: multiple angiomyolipomas in 18.4%, cysts in 18.4% and angiomyolipomas plus cysts in 7.9%. Renal cysts were detected at a median age of 3.1 years and angiomyolipomas at 10.7 years. Two patients have required invasive procedures due to symptomatic large cysts, 5 developed proteinuria and 3 arterial hypertension. None presented renal carcinoma neither evolved to chronic renal failure. CONCLUSIONS. Our findings confirm a high rate of renal involvement in children with tuberous sclerosis, thus we emphasize periodic renal surveillance of these patients in order to diminish the morbidity of this disease. PMID- 19809766 TI - [Juvenil idiopathic arthritis. Part 1: diagnosis, pathogenesis and clinical manifestations]. AB - Juvenile idiopathic arthritis is not a single disease and constitutes an heterogeneous group of illnesses or inflammatory disorders. This new nomenclature encompasses different disease categories, each of which has different presentation, clinical signs, symptoms, and outcome. The cause of the disease is still unknown but both environmental and genetic factors seem to be related to its pathogenesis. Is the most common chronic rheumatic disease in children and an important cause of short-term and long-term disability. In this article, clinical manifestation, new classification and approach to diagnosis are reviewed. PMID- 19809767 TI - [Whooping cough: clinical and epidemiological characteristics of 20 confirmed cases of the Paediatric Hospital of Misiones province]. AB - Pertussis or whooping cough is a respiratory disease that has emerged in recent years in several countries including Argentina. The aim is to retrospectively describe clinical and epidemiological characteristics of 20 patients in the Pediatric Hospital of Misiones with confirmed diagnosis of whooping cough, according to criteria set by WHO and CDC, during the years 2005 and 2006. The median age was 4 months, 13 were male (65%). Fourteen patients (70%) were younger than 6 months and 9 (45%) younger than 3 months. All had cough (average duration of 7.6 days), 5 (25%) paroxysmal cough and 1 (5%) apnea and cyanosis. Two children died. Sixteen (80%) had not the three doses of vaccine quadruple, 7 children (35%) were younger than 3 months and had no dose. The disease remains a public health problem affecting not only children but also adults. PMID- 19809768 TI - [Chronic autoimmune urticaria: treatment with omalizumab]. AB - We report the case of a child with diagnosis of chronic urticaria/angioedema and its evolution upon omalizumab treatment. CASE REPORT: Our patient is a 12-years old female who suffered for 14 months severe chronic urticaria/angioedema. She had a poor response to the highest doses of combined therapy with 3 antihistamines, steroids and anti-leukotrienes and great impairment of her quality of life. An autologous serum skin test was positive until 1:100 dilutions, leading to the diagnosis of chronic autoimmune urticaria. Due to the lack of response to treatment, therapy with omalizumab was administered. A notable reduction in symptoms toward the third dose was observed. After 12 months of this treatment, the patient is asymptomatic and has a negative autologous serum test. CONCLUSION: Omalizumab could be a therapeutic option for patients with autoimmune urticaria unresponsive to other treatments. PMID- 19809769 TI - [Amitraz poisoning]. AB - Poisoning due to amitraz together with its solvent xilene, is an unusual condition although may be increasing in rural areas where it is used as insecticide-ectoparasiticide.1-3 At present, there is scare references to orient physicians concerning its handling in childhood. We present the case of a 2-year old boy who suffered an accidental intake of amitraz and was admitted into our Pediatric Intensive Care Unit requiring mechanical ventilation. We consider the usefulness of informing the medical community about this case so as to be aware of this rare kind of poisoning in our community. PMID- 19809770 TI - [Cow's milk protein allergy: proposed guidelines for the management of children with cow's milk protein allergy]. AB - Cow's milk allergy is a growing concern in the practice of pediatrics. The impression of an increasing incidence, similar to what has been reported in other latitudes, has determined the need for guidelines to help in the diagnosis and treatment of this disease. A group of pediatric specialists met to discuss the "state of the art" and propose local guidelines to deal with cow's milk allergy. The aim has been to contribute in the understanding of the pathophysiology, environmental factors, and clinical expressions of this problem, and help pediatricians in the overall management. PMID- 19809771 TI - [Chronic endemic regional hydroarsenicism: a challenge for diagnosis and prevention]. AB - Arsenic (As) is a semimetal that is widely distributed in nature, in water and soil. In Argentine, the contamination of both waterways and groundwater represents the main environmental problem caused by this element. Chronic As poisoning is known as Chronic endemic regional hydroarsenicism (C.E.R.HA.). Long term exposure to low concentrations of the element from the prenatal period onward results in the well-known symptoms of chronic As poisoning. CERHA develops progressively, compromising different organs and systems, most importantly the skin. One of the most important complications of CERHA is de development of neoplasias, mainly skin tumors. Childhood environmental health is a challenge in the new millennium and health care professionals play a fundamental role in the protection against environmental hazards such as chronic arsenic poisoning. PMID- 19809772 TI - Synergistic action of exogenous salicylic acid and arbuscular mycorrhizal fungus colonization in Avena nuda seedlings in response to NO(2) exposure. AB - Colonization of arbuscular mycorrhizal fungi Glomus mosseae or exogenous salicylic acid (SA) treatment can increase Avena nuda plant tolerance to elevated NO(2) exposure. The combination of the two factors, namely application of SA to the mycorrhizal plants, further promoted NO(2) tolerance, as indicated by an alleviated plant biomass decrease compared to the respective treatment. The analysis of antioxidant capacity, redox status and photon energy utilization showed that the increased NO(2) tolerance in the treated plants may be associated, at least in part, with scavenging reactive oxygen species, maintaining CO(2) assimilated rate and reducing conditions in cells. PMID- 19809773 TI - Dissipation of mefenoxam residue in watermelon and soil under field conditions. AB - Mefenoxam is the systemic phenylamide fungicide, which is widely used in controlling phytophthora disease of tomato, cucumber, pepper and watermelon, etc. The dissipation behaviour of mefenoxam residues in watermelons and soils was studied. The whole watermelon, melon flesh and soil matrices of mefenoxam were analyzed by GC-NPD. At three different spiking levels mean recoveries and relative standard deviation from spiked samples in six replicated experiments for each matrix were in the range 89.6-98.2% and 1.5-8.1%, respectively. Under field conditions, mefenoxam dissipation rate was found to be faster in the whole watermelons than in the soils. The results showed that the half lives in whole watermelon and soil from Beijing were 3.9 and 10.0 days, respectively, and the half lives in whole watermelon and soil from Shanxi were 3.7 and 28.4 days. PMID- 19809774 TI - Nitrates and herbicides cause higher mortality than the traditional organic fertilizers on the grain beetle, Tenebrio molitor. AB - We examined experimentally whether fertilizers or herbicides commonly used by farmers affect mortality of the adult grain beetle Tenebrio molitor. After a period of 4 weeks in direct contact with all treatments, a higher percentage of mortality occurred in contact with nitrates than with pig manure or turkey litter. Herbicides (a mixture of glyphosate and 2,4-D: ) caused 100% mortality. Our results also indicate that more beetles escaped from the herbicides and nitrate treatments than from the others, suggesting some kind of behavioural avoidance of toxic environments. The traditional organic fertilizers appear to be less toxic than inorganic fertilizers for Tenebrio molitor. PMID- 19809775 TI - Hip fractures in Italy: 2000-2005 extension study. AB - SUMMARY: A total of 507,671 people > or =65 experienced hip fractures between 2000 and 2005. In 2005, 94,471 people > or =65 were hospitalized due to hip fractures, corresponding to a 28.5% increase over 6 years. Most fractures occurred in patients > or =75 (82.9%; n = 420,890; +16% across 6 years), particularly in women (78.2%; n = 396,967). INTRODUCTION: We aimed to analyze incidence and costs of hip fractures in Italy over the last 6 years. METHODS: We analyzed the national hospitalization and DRG databases concerning fractures occurred in people > or =65 between 2000 and 2005. RESULTS: A total of 507,671 people > or =65 experienced hip fractures across 6 years, resulting in about 120,000 deaths. In year 2005 94,471 people aged > or =65 were hospitalized due to hip fractures, corresponding to a 28.5% increase over 6 years. The majority of hip fractures occurred in patients > or =75 (82.9%; n = 420,890; +16% across 6 years) and particularly in women (78.2%; n = 396,967). Among women, 84.2% of fractures (n = 334,223; +28.0% over 6 years) were experienced by patients > or =75, which is known to be the age group with the highest prevalence of osteoporosis, accounting for 68.6% of the overall observed increase in the total number of fractures. Hip fractures in men > or =75 increased by 33.1% (up to 16,540). Hospitalization costs increased across the six examined years (+36.1%) reaching 467 million euros in 2005, while rehabilitation costs rose up to 531 million in the same year. CONCLUSIONS: Hip fractures of the elderly are increasing and represent a major health problem in industrialized countries such as Italy. PMID- 19809776 TI - Detecting early bone changes using in vivo micro-CT in ovariectomized, zoledronic acid-treated, and sham-operated rats. AB - SUMMARY: This study monitored in vivo the effect on bone microarchitecture of initiating antiresorptive treatment with zoledronic acid in rats at 2 weeks following ovariectomy, an early phase at which major degenerative bone changes have been found to occur. The treatment still facilitated the full reversal of cancellous bone loss in rat tibia, highlighting the importance of the time point of initiation of antiresorptive treatment. INTRODUCTION: Injection of zoledronic acid in rats at time of ovariectomy has been found to fully preserve tibial bone microarchitecture over time, whereas injection at 8 weeks after ovariectomy has shown partial bone recovery. This study investigated the effect on microarchitecture of initiating antiresorptive treatment in the early phase following ovariectomy, at 2 weeks, a time point at which major degenerative changes in the bone have been found to occur. METHODS: Female Sprague-Dawley rats were divided into ovariectomized group, ovariectomized group treated with zoledronic acid, and sham-operated group. In vivo micro-CT scanning of rat tibiae and morphometric analysis were performed at 0, 2, 4, 8, and 12 weeks after ovariectomy, with zoledronic acid treatment beginning 2 weeks after ovariectomy. Data were first analyzed with repeated measures analysis of variance (longitudinal study design) and then without repeated measures (cross-sectional study design). RESULTS: The ovariectomized group demonstrated dramatic bone loss, first detected at week 2. Conversely, at week 4, the zoledronic acid-treated group returned microstructural parameters to baseline values. Remarkable increases in bone parameters were found after 6 weeks of treatment and maintained similar to sham group until the end. The longitudinal study design provided earlier detection of bone changes compared to the cross-sectional study design. CONCLUSIONS: Treatment with zoledronic acid as late as 2 weeks after ovariectomy still facilitates the full reversal of cancellous bone loss in the rat tibia. PMID- 19809777 TI - Radiologic observation: repair of focal bone erosions after humanized antitumor necrosis factor antibody adalimumab therapy in a patient with rheumatoid arthritis. AB - Erosions of the bone and cartilage are considered as a cardinal feature of rheumatoid arthritis (RA) leading to joint destruction and functional limitations. This report is a radiologic observation of distinguishable bone erosion repair with concomitant increase of radio-opacity of trabecular bones after 15 months of antitumor necrosis factor (TNF) antibody adalimumab therapy in a 26-year-old woman with RA resistant to conventional therapy. Although the introduction of anti-TNF antibodies has contributed to the slowing and arrest of RA progression, destruction itself has been generally acknowledged as an irreversible process with little hope (only 1.8% of erosions) in resolution. Our patient's dramatic structural retrieval is a discrepant, yet notable case pivoting the previous belief on the skeletal restoration capacity of anti-TNF antibody into a circulating cytokine-dependent manner. PMID- 19809779 TI - The functional anatomy of the cricothyroid joint. AB - Postoperative long-term treatment problems of male-to-female transsexuals can be caused by the underestimated mobility of the cricothyroid joint. One hundred cadaveric larynges were analysed to reveal the morphological and functional anatomy of the CTJ focusing on possible horizontal and vertical gliding movements. Furthermore, the distance of the cricothyroid space for rotational movements was measured. The articular surface of the cricothyroid joint showed three different possibilities: a well-defined facet (Type 1 61%), no definable facet (Type 2 22%) or a flat surface with or without a tiny protuberance (Type 3 17%). Side different statements and intraindividual differences between male and female specimens were included. These different types affect horizontal and vertical gliding movements, besides rotational movements. The mobility between Type 1 and the others was highly significant (P < 0.001). Regarding different motions in the cricothyroid joint, rotational movements caused significant elongation of the vocal folds, which should be taken into consideration for phonosurgical methods. PMID- 19809778 TI - Non-Hodgkin's lymphoma in systemic sclerosis: case and literature review. AB - The occurrence of non-Hodgkin's lymphoma in systemic sclerosis is an uncommon event. In our scleroderma cohort, a case of primary gastric B-cell lymphoma was diagnosed in 2007. The patient was a 45-year-old woman suffering from late systemic sclerosis sine scleroderma in whom non-Hodgkin's lymphoma presented with progressive weight loss, later with gastrointestinal symptoms. Subsequently, we retrospectively analyzed the charts of 251 systemic sclerosis patients consecutively admitted to our Unit from 2000 to 2008 to search for other non Hodgkin's lymphoma cases (prevalence, 0.49%). Then we performed a Pubmed search for "systemic sclerosis & non-Hodgkin's lymphoma," limited to the English language. Twenty detailed cases of such an association were found, pointing out the following: non-Hodgkin's lymphoma seems to be associated to old age, female sex, diffuse cutaneous subset and early disease; B-cell lymphoma subtypes are the majority; the interval between systemic sclerosis and lymphoma onset is usually short; systemic sclerosis could present as a paraneoplastic syndrome in some cases. We concluded that, although rare, the association of systemic sclerosis and non-Hodgkin's lymphoma may not be coincidental and the clinician should be aware of the risk for lymphoproliferative disorders in scleroderma patients. This is the first description of non-Hodgkin's lymphoma in systemic sclerosis sine scleroderma. The insidious onset of gastric lymphomas, mimicking the most common features of gastrointestinal involvement in systemic sclerosis is underlined. PMID- 19809780 TI - [Perspectives for professionally sufficient and adequate care of ageing patients]. AB - This contribution deals with the question of how existing health care systems can be sufficiently and adequately adapted to ageing patients in general, and to patients with special needs for nursing care and assistance. Simultaneously, it advocates explicit consideration of the dimensions of social inequality and health hazards related to biographical factors. It demonstrates that health care involves much more than just adequate medical care in the event of illness. The reasons that change is needed are given and reform options for relevant segments of health care (such as medical practitioners, hospital care, health care for patients in need of nursing care, palliative care, prevention and rehabilitation) are presented. Special emphasis is placed on standards and guidelines relating to old age morbidity and integrated health care systems as well as rehabilitation, prevention and health promotion. A key role is attributed to the qualification of health services staff. More intensive research into health care and health services in respect of the elderly is also advocated. PMID- 19809781 TI - Magnetic resonance imaging of hepatocellular carcinoma: a pictorial review of novel insights into pathophysiological features revealed by magnetic resonance imaging. AB - Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide. Magnetic resonance (MR) imaging is one of the most powerful modalities for the assessment of HCC with sufficient sensitivity and specificity. In addition to its capacity for lesion detection, MR imaging delineates some unique in vivo pathophysiological features of tumors, which cannot be assessed by other modalities. Chemical shift imaging may depict steatosis of the tumor. Dynamic contrast-enhanced MR imaging is the most powerful tool to assess the vascularity of the tumor, which is closely related to malignant transformation in hepatocarcinogenesis. Diffusion-weighted imaging illustrates the cellularity of the tumor. Super-paramagnetic iron oxide, a liver-specific MR contrast agent accumulating in Kupffer cells, enables detection of the hepatocellular architecture in the lesion. Recently, a new liver-specific MR contrast agent, gadoxetic acid [gadolinium-ethoxybenzyl (Gd-EOB)-diethylenetriaminopentoacetic acid (DTPA)], has been introduced for clinical imaging. Gd-EOB-DTPA has a significant impact on the imaging of HCC, with potential capacity for the concurrent assessment of vascularity of the tumor and hepatocellular-specific properties within the tumor. Understanding the characteristics of MR imaging methods and contrast agents is essential for the optimal diagnosis and characterization of HCCs. PMID- 19809782 TI - Development of nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) after pancreaticoduodenectomy: proposal of a postoperative NAFLD scoring system. AB - BACKGROUND: The main etiology of NAFLD and NASH after pancreatic resection is still unclear, and the therapeutic strategy has yet to be established. The focus of this review is how predict and prevent NAFLD/NASH after pancreaticoduodenectomy. METHODS: From April 2005 to October 2008, 54 patients who underwent pancreaticoduodenectomy in our institution were enrolled in this study. From the pre-, intra- and postoperative risk factors, we identified the most influential risk factors of postoperative NAFLD by uni- and multivariate analyses. Moreover, a postoperative NAFLD scoring system was proposed based on these risk factors. RESULTS: The incidence of postoperative NAFLD was 37.0% (20/54). Of these, 10% (2/20) of patients were diagnosed as having NASH by percutaneous liver biopsy. By multivariate analysis, pancreatic adenocarcinoma (p < 0.05), pancreatic resection line (p < 0.01) and postoperative diarrhea (p < 0.01) were identified as the most influential factors concerning postoperative NAFLD. Based on these results, we proposed a postoperative NAFLD scoring system (0-10) and evaluated the correlation between the score and decreasing rates of CT values, revealing a significant correlation (r = 0.829 p < 0.001). The prevalence of postoperative NAFLD in the patients with our scores of 0-3, 4-6 and 7-10 points was 0 (0/22), 35 (6/17) and 93% (14/15), respectively. CONCLUSIONS: In conclusion, NAFLD develops frequently in patients who undergo PD, and some patients even progress to NASH. A postoperative NAFLD scoring system makes it possible to predict the occurrence of NAFLD after PD, and aggressive nutrition support is needed for patients with high scores. PMID- 19809783 TI - Analysis of T-cell receptor usage in myeloperoxidase--antineutrophil cytoplasmic antibody-associated renal vasculitis. AB - BACKGROUND: Bacterial superantigens produced by Staphylococcus aureus may be associated with the onset of proteinase-3 antineutrophil cytoplasmic antibody (PR3-ANCA)-associated vasculitis, including Wegener's granulomatosis. We investigated T-cell subsets to assess the superantigens present in patients with myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA)-associated vasculitis. METHODS: Peripheral-blood mononuclear cells (PBMC) obtained from 40 normal controls and ten patients with MPO-ANCA-associated vasculitis were stained with fluorescence-labeled monoclonal antibodies against T-cell markers, including 17 variable regions of T-cell receptor beta-chains (TCR-Vbeta) and were then analyzed using flow cytometry. RESULTS: Among PBMCs, the percentage of CD3(+) cells from patients with MPO-ANCA-associated vasculitis was significantly lower than that from normal controls, but there were no differences between the two groups in the percentage of CD19(+) cells or CD16(+) cells. Although there were no differences regarding the overall percentage of CD4(+) cells between the two groups, the percentage of CD4(+)CD45RO(+) cells in patients with MPO-ANCA associated vasculitis was significantly higher than that in normal controls, and percentages of CD4(+)CD45RO(+)HLA-DR(+) and CD4(+)CD45RO(+)CD62L(low) cells in patients with MPO-ANCA-associated vasculitis were also significantly increased. There was no significant difference between the two groups in terms of the usage of the 17 different TCR-Vbeta regions. CONCLUSION: There was no difference in bacterial superantigens between controls and MPO-ANCA-associated vasculitis patients because of the absence of specific usage of TCR-Vbeta regions. Given the elevated levels of memory T cells, conventional antigens rather than superantigens may be associated with the pathogenesis of MPO-ANCA-associated vasculitis. PMID- 19809784 TI - Exercise-induced acute kidney injury with reversible posterior leukoencephalopathy syndrome. AB - We report a case of a 17-year-old boy suffering from severe loin pain and oliguric acute kidney injury after strenuous exercise, with slightly elevated serum myoglobin and creatinine phosphokinase. Exercise-induced acute kidney injury (AKI) was diagnosed. We started intermittent hemodialysis thrice a week from the admission day. Four days later, he temporally lost consciousness, followed by visual agnosia and general clonic seizure. T2-weighted brain magnetic resonance images (MRI) showed multiple areas of increased signal intensity in the subcortical white matter, predominantly in the cerebrum of the posterior and parietal lobes and in the cerebellum. Clinical symptoms improved without sequelae. Follow-up MRI 1 month later showed complete resolution of the signals, and he was diagnosed with reversible posterior leukoencephalopathy syndrome (RPLS). This is the first reported case of exercise-induced AKI associated with RPLS. Vasoconstriction and endothelial dysfunction are considered as the common etiology of these diseases. PMID- 19809785 TI - Focal segmental glomerulosclerosis in a patient with isolated ACTH deficiency and reversible hypothyroidism. AB - A 23-year-old man was admitted to our hospital for fatigue, anorexia, proteinuria, and peripheral edema. Proteinuria was first pointed out at the age of 15, but no further studies were performed. Six years prior to admission, the patient noted becoming easily fatigued. Laboratory tests on admission showed marked peripheral eosinophilia (29.2%, count: 1,071/microL) and proteinuria. Endocrinological studies revealed isolated adrenocorticotropic hormone deficiency with primary hypothyroidism, but a lack of autoimmune thyroiditis. Renal biopsy showed focal segmental glomerulosclerosis. Hydrocortisone therapy (30 mg/day) for isolated adrenocorticotropic hormone deficiency resulted in marked improvement of adrenal and thyroid functions, and amelioration of proteinuria (from 2.8 to 1.0 g/day) over a two-month period. Renal function remains normal at five years after the start of hydrocortisone treatment. The findings suggest that both hydrocortisone therapy and normalized thyroid hormone status played a pivotal role in the improvement of proteinuria associated with focal segmental glomerulosclerosis. PMID- 19809786 TI - A quantitative comparison of radiosurgical treatment parameters in vestibular schwannomas: the Leksell Gamma Knife Perfexion versus Model 4C. AB - PURPOSE: The world's first Gamma Knife Perfexion (PFX)was installed in Marseille in July 2006. The aim of this study was to investigate the impact of the PFX technology on the quality of dose planning for vestibular schwannomas (VS). METHODS: When the PFX was first introduced, a comparative randomized prospective study of 200 patients was conducted.Seventy-eight of the 200 patients in that study had VS, of whom 38 were randomized to treatment with the Gamma Knife Model 4C (group 4C) and 40 were randomized to treatment with PFX (group P1). The authors also incorporated a matched group of 40 patients with VS consecutively treated with PFX after the initial learning curve period (group P2). Dose planning was compared and evaluated by measuring the conformity index (CI), selectivity index (SI), gradient index(GI), energy index (EI), unit isocenters (UI) and cochlear dose. Patients were also stratified into subgroups according to target volume (> or = 0.5 ml). RESULTS: In the whole population, CI, EI and cochlear dose were significantly better in group P2 (CI=0.917, EI=1.35,cochlear dose=3.55) than in group 4C (CI=0.864, EI=1.27,cochlear dose=5.10). In the subgroup of lesions > or = 0.5 ml, CI,GI, EI, UI and cochlear dose in group P2 (CI=0.929, GI=2.67, EI=1.37, UI=10.6, cochlear dose=3.55) were significantly better than in group 4C (CI=0.874, GI=2.85, EI=1.30, UI=14.5, cochlear dose=5.10). CONCLUSIONS: The investigation of the dose planning capabilities of the PFX on a cohort of VS demonstrates a better conformity and energy distribution, with better cochlear sparing and without any particular drawback. In addition,there is an improvement in peripheral dose gradient in larger lesions. Further clinical studies will be required before drawing any conclusions about the clinical benefit achieved by these dose planning improvements. PMID- 19809787 TI - Reversible posterior leukoencephalopathy syndrome in childhood: report of nine cases and review of the literature. AB - Reversible posterior leukoencephalopathy syndrome (RPLS) is recently described disorder with typical radiological findings in the posterior regions of the cerebral hemisphere and cerebellum. Its clinical symptoms include headache, decreased alertness, mental abnormalities, such as confusion, diminished spontaneity of speech, and changed behavior ranging from drowsiness to stupor, seizures, vomiting and abnormalities of visual perception like cortical blindness. RPLS is caused by various heterogeneous factors, the commonest being hypertension, followed by non-hypertensive causes such as eclampsia, renal diseases and immunosuppressive therapy. We presented nine patients with RPLS who had primary diagnoses such as acute post-streptococcal glomerulonephritis, idiopathic hypertension, the performing of intravenous immunoglobulin for infection with crescentic glomerulonephritis, erythrocyte transfusion for severe iron deficiency, L: -asparaginase treatment for acute lymphoblastic leukemia and performing of granulocyte-colony stimulating factor for ulcerative colitis due to neutropenia. Early recognition of RPLS as complication during different diseases and therapy in childhood may facilitate precise diagnosis and appropriate treatment. PMID- 19809788 TI - 9 november 2009: in-depth oral presentations and oral communications. PMID- 19809789 TI - 10 november 2009: in-depth oral presentations and oral communications. PMID- 19809790 TI - 11 november 2009: in-depth oral presentations and oral communications. PMID- 19809791 TI - Poster session. PMID- 19809793 TI - Distribution, cellular localization, and therapeutic potential of the tumor associated antigen Ku70/80 in glioblastoma multiforme. AB - Antibodies specifically targeting tumor-associated antigens have proved to be important tools in the treatment of human cancer. A desirable target antigen should be unique to tumor cells, abundantly expressed, and readily available for antibody binding. The Ku70/80 DNA-repair protein is expressed in the nucleus of most cells; it is, however, also present on the cell surface of tumor cell lines, and antibodies binding Ku70/80 at the cell surface were recently shown to internalize into tumor cells. To evaluate the potential of Ku70/80-antigen as a therapeutic target for immunotoxins in glioblastoma multiforme, we investigated binding and localization of Ku70/80-specific antibodies in tissue samples from glioblastomas and normal human brains, and in glioma cell cultures. Furthermore, the internalization and drug-delivery capacity were evaluated by use of immunotoxicity studies. We demonstrate that Ku70/80 is localized on the cell plasma membrane of glioma cell lines, and is specifically present in human glioblastoma tissue. Antibodies bound to the Ku70/80 antigen on the cell surface of glioma cells were found to internalize via endocytosis, and shown to efficiently deliver toxins into glioblastoma cells. The data further imply that different antibodies directed against Ku70/80 possess different abilities to target the antigen, in relation to its presentation on the cell surface or intracellular localization. We conclude that Ku70/80 antigen is uniquely presented on the plasma membrane in glioblastomas, and that antibodies specific against the antigen have the capacity to selectively bind, internalize, and deliver toxins into tumor cells. These results imply that Ku70/80 is a potential target for immunotherapy of glioblastoma multiforme. PMID- 19809794 TI - Enhanced death signaling in ozone-exposed ischemic-reperfused hearts. AB - Although numerous advancements made in the field of human health have resulted in reduced deaths due to cardiovascular diseases (CVD), many patients with cardiac disease show no established risk. Therefore, other unknown factors may be responsible for the pathophysiology of CVD. Out of 350,000 sudden cardiac deaths each year in the United States, 60,000 deaths have been related to air pollution, suggesting a detrimental role of environmental pollutants in the development of CVD. The present study tested our hypothesis that chronic ozone exposure enhances the sensitivity to ischemia-reperfusion (I/R) injury in isolated perfused hearts. Sprague-Dawley rats were continuously exposed for 8 h/day for 28 and 56 days to filtered air or 0.8 ppm ozone. Isolated hearts were subjected to 30 min of global ischemia followed by 60 min of reperfusion. Cardiac function after I/R measured as left ventricular developed pressure (LVDP), +dP/dt, -dP/dt, and left ventricular end diastolic pressure (LVEDP) was significantly decreased and increased respectively in ozone-exposed I/R hearts compared to I/R hearts exposed to filtered air. The enhanced sensitivity to I/R injury upon ozone exposure was associated with increased myocardial TNF-alpha levels and lipid peroxidation and decreased myocardial activities of superoxidase dismutase (SOD) and IL-10. These data suggest that ozone-induced sensitivity to myocardial I/R injury may be due to promoting levels of oxidative stress as well as inflammatory mediators. PMID- 19809792 TI - The exercising heart at altitude. AB - Maximal cardiac output is reduced in severe acute hypoxia but also in chronic hypoxia by mechanisms that remain poorly understood. In theory, the reduction of maximal cardiac output could result from: (1) a regulatory response from the central nervous system, (2) reduction of maximal pumping capacity of the heart due to insufficient coronary oxygen delivery prior to the achievement of the normoxic maximal cardiac output, or (3) reduced central command. In this review, we focus on the effects that acute and chronic hypoxia have on the pumping capacity of the heart, particularly on myocardial contractility and the molecular responses elicited by acute and chronic hypoxia in the cardiac myocytes. Special emphasis is put on the cardioprotective effects of chronic hypoxia. PMID- 19809795 TI - The NMR restraints grid at BMRB for 5,266 protein and nucleic acid PDB entries. AB - Several pilot experiments have indicated that improvements in older NMR structures can be expected by applying modern software and new protocols (Nabuurs et al. in Proteins 55:483-186, 2004; Nederveen et al. in Proteins 59:662-672, 2005; Saccenti and Rosato in J Biomol NMR 40:251-261, 2008). A recent large scale X-ray study also has shown that modern software can significantly improve the quality of X-ray structures that were deposited more than a few years ago (Joosten et al. in J. Appl Crystallogr 42:376-384, 2009; Sanderson in Nature 459:1038-1039, 2009). Recalculation of three-dimensional coordinates requires that the original experimental data are available and complete, and are semantically and syntactically correct, or are at least correct enough to be reconstructed. For multiple reasons, including a lack of standards, the heterogeneity of the experimental data and the many NMR experiment types, it has not been practical to parse a large proportion of the originally deposited NMR experimental data files related to protein NMR structures. This has made impractical the automatic recalculation, and thus improvement, of the three dimensional coordinates of these structures. We here describe a large-scale international collaborative effort to make all deposited experimental NMR data semantically and syntactically homogeneous, and thus useful for further research. A total of 4,014 out of 5,266 entries were 'cleaned' in this process. For 1,387 entries, human intervention was needed. Continuous efforts in automating the parsing of both old, and newly deposited files is steadily decreasing this fraction. The cleaned data files are available from the NMR restraints grid at http://restraintsgrid.bmrb.wisc.edu . PMID- 19809796 TI - Polymorphisms within insulin-degrading enzyme (IDE) gene determine insulin metabolism and risk of type 2 diabetes. AB - Insulin-degrading enzyme (IDE) is the ubiquitously expressed major enzyme responsible for insulin degradation. Insulin-degrading enzyme gene is located on chromosome region 10q23-q25 and exhibits a well-replicated peak of linkage with type 2 diabetes (T2DM). Several genetic association studies examined IDE gene as a susceptibility gene for T2DM with controversial results. However, pathophysiological mechanisms involved have remained elusive. We verified associations of two IDE polymorphisms (rs1887922 and rs2149632) with T2DM risk in two independent German cohorts and evaluated in detail the association of common variants with insulin metabolism and glycemic traits. We confirmed previously published findings for diabetes-associated rs1887922 and rs2149632 in the European Prospective Investigation into Cancer and Nutrition-Potsdam cohort (n = 3049; RR 1.26, p = 0.003 and RR 1.33, p < 0.0001 for additive model). Haplotypes which carried one risk allele of rs2149632 or two risk alleles of both studied IDE SNPs also demonstrated a strong association with increased T2DM risk in this cohort (p = 0.001 and p < 0.0001, respectively). However, we found no significant T2DM association in the cross-sectional metabolic syndrome Berlin-Potsdam cohort (n = 1026). In nondiabetic subjects (NGT+IFG/IGT; n = 739), we found an association of rs2149632 with impaired glucose-derived insulin secretion and a trend to decreased insulin sensitivity for rs1887922. In the NGT subjects (n = 440), the association with decreased insulin secretion for rs2149632 remain significant, and the association with decreased hepatic insulin degradation for rs1887922 were observed additionally. This study validates and confirms the association of IDE polymorphisms with T2DM risk in the prospective German cohort and provides novel evidence of influences of IDE genetic variants on insulin metabolism. PMID- 19809797 TI - Diverse roles for protein kinase C delta and protein kinase C epsilon in the generation of high-fat-diet-induced glucose intolerance in mice: regulation of lipogenesis by protein kinase C delta. AB - AIMS/HYPOTHESIS: This study aimed to determine whether protein kinase C (PKC) delta plays a role in the glucose intolerance caused by a high-fat diet, and whether it could compensate for loss of PKCepsilon in the generation of insulin resistance in skeletal muscle. METHODS: Prkcd (-/-), Prkce (-/-) and wild-type mice were fed high-fat diets and subjected to glucose tolerance tests. Blood glucose levels and insulin responses were determined during the tests. Insulin signalling in liver and muscle was assessed after acute in vivo insulin stimulation by immunoblotting with phospho-specific antibodies. Activation of PKC isoforms in muscle from Prkce (-/-) mice was assessed by determining intracellular distribution. Tissues and plasma were assayed for triacylglycerol accumulation, and hepatic production of lipogenic enzymes was determined by immunoblotting. RESULTS: Both Prkcd (-/-) and Prkce (-/-) mice were protected against high-fat-diet-induced glucose intolerance. In Prkce (-/-) mice this was mediated through enhanced insulin availability, while in Prkcd (-/-) mice the reversal occurred in the absence of elevated insulin. Neither the high-fat diet nor Prkcd deletion affected maximal insulin signalling. The activation of PKCdelta in muscle from fat-fed mice was enhanced by Prkce deletion. PKCdelta deficient mice exhibited reduced liver triacylglycerol accumulation and diminished production of lipogenic enzymes. CONCLUSIONS/INTERPRETATION: Deletion of genes encoding isoforms of PKC can improve glucose intolerance, either by enhancing insulin availability in the case of Prkce, or by reducing lipid accumulation in the case of Prkcd. The absence of PKCepsilon in muscle may be compensated by increased activation of PKCdelta in fat-fed mice, suggesting that an additional role for PKCepsilon in this tissue is masked. PMID- 19809799 TI - Suppression of the TRIF-dependent signaling pathway of toll-like receptors by isoliquiritigenin in RAW264.7 macrophages. AB - Toll-like receptors (TLRs) play an important role in host defense by sensing invading microbial pathogens and initiating innate immune responses. The stimulation of TLRs by microbial components triggers the activation of myeloid differential factor 88 (MyD88)- and toll-interleukin-1 receptor domain-containing adapter inducing interferon-beta (TRIF)-dependent downstream signaling pathways. Isoliquiritigen in (ILG), an active ingredient of Licorice, has been used for centuries to treat many chronic diseases. ILG inhibits the MyD88-dependent pathway by inhibiting the activity of inhibitor-kappaB kinase. However, it is not known whether ILG inhibits the TRIF-dependent pathway. To evaluate the therapeutic potential of ILG, we examined its effect on signal transduction via the TRIF-dependent pathway of TLRs induced by several agonists. ILG inhibited nuclear factor-kappaB and interferon regulatory factor 3 activation induced by lipopolysaccharide or polyinosinic-polycytidylic acid. ILG inhibited the lipopolysaccharide-induced phosphorylation of interferon regulatory factor 3 as well as interferon-inducible genes such as interferon inducible protein-10, and regulated activation of normal T-cell expressed and secreted (RANTES). These results suggest that ILG can modulate TRIF-dependent signaling pathways of TLRs, leading to decreased inflammatory gene expression. PMID- 19809800 TI - Characterization of putative capsaicin synthase promoter activity. AB - Capsaicin is a very important secondary metabolite that is unique to Capsicum. Capsaicin biosynthesis is regulated developmentally and environmentally in the placenta of hot pepper. To investigate regulation of capsaicin biosynthesis, the promoter (1,537 bp) of pepper capsaicin synthase (CS) was fused to GUS and introduced into Arabidopsis thaliana (Col-0) via Agrobacterium tumefaciens to produce CSPRO::GUS transgenic plants. The CS was specifically expressed in the placenta tissue of immature green fruit. However, the transgenic Arabidopsis showed ectopic GUS expressions in the leaves, flowers and roots, but not in the stems. The CSPRO activity was relatively high under light conditions and was induced by both heat shock and wounding, as CS transcripts were increased by wounding. Exogenous capsaicin caused strong suppression of the CSPRO activity in transgenic Arabidopsis, as demonstrated by suppression of CS expression in the placenta after capsaicin treatment. Furthermore, the differential expression levels of Kas, Pal and pAmt, which are associated with the capsaicinoid biosynthetic pathway, were also suppressed in the placenta by capsaicin treatment. These results support that capsaicin, a feedback inhibitor, plays a pivotal role in regulating gene expression which is involved in the biosynthesis of capsaicinoids. PMID- 19809798 TI - Insights into the critical role of NADPH oxidase(s) in the normal and dysregulated pancreatic beta cell. AB - It is now widely accepted that reactive oxygen species (ROS) contribute to cell and tissue dysfunction and damage in diabetes. The source of ROS in the insulin secreting pancreatic beta cells has traditionally been considered to be the mitochondrial electron transport chain. While this source is undoubtedly important, we fully describe in this article recent information and evidence of NADPH oxidase-dependent generation of ROS in pancreatic beta cells and identify the various isoforms that contribute to O(2)(*-) and H(2)O(2) production in various conditions. While glucose-stimulated ROS generation may be important for acute regulation of insulin secretion, at higher levels ROS may disrupt mitochondrial energy metabolism. However, ROS may alter other cellular processes such as signal transduction, ion fluxes and/or cell proliferation/death. The various beta cell isoforms of NADPH oxidase (described in this review) may, via differences in the kinetics and species of ROS generated, positively and negatively regulate insulin secretion and cell survival. PMID- 19809802 TI - Electroconvulsive therapy for refractory status epilepticus: a case series. AB - BACKGROUND: Status epilepticus refractory to conventional anti-epileptic drugs typically has a poor prognosis, but patients may recover well if seizures can be stopped. Case reports suggest that electroconvulsive therapy (ECT) may stop seizures in patients with refractory status epilepticus, and we sought to examine its effectiveness in a series of patients. METHODS: Three consecutive patients with refractory status epilepticus at our institution were treated with ECT after other therapies had failed. RESULTS: ECT stopped seizures in 2 of 3 patients. One patient had complete neurological recovery; the other was left with mild cognitive impairment and epilepsy, but returned to independent living. CONCLUSION: ECT may be an effective therapy for refractory status epilepticus and warrants further study for this indication. PMID- 19809803 TI - Solid cell nests in Hashimoto's thyroiditis sharing features with papillary thyroid microcarcinoma. AB - Solid cell nests (SCN) associated with Hashimoto's thyroiditis may show some atypical nuclear features including prominent nuclear grooves, enlarged overlapping nuclei and nuclear clearing. These features are sometimes mistaken for papillary thyroid microcarcinomas especially when the SCN are numerous. We reviewed SCN associated with Hashimoto's thyroiditis in 12 patients selected from 1,420 archival routinely processed formalin-fixed, paraffin-embedded thyroid specimens of Hashimoto's thyroiditis in which there was more than ten SCN per slide. In addition to the atypical nuclear features, there was a distinct eosinophilic basement membrane surrounding the SCN. Immunohistochemical analysis showed that the SCN were strongly positive for p63, stained weakly for TTF-1 and were negative for thyroglobulin, HBME-1, and calcitonin. This was compared to papillary thyroid microcarcinomas which were strongly positive for thyroglobulin, TTF-1, HBME-1, and variably positive for p63, while calcitonin and chromogranin were negative. These histological and immunophenotypic features can be used to distinguish SCN from papillary thyroid microcarcinomas associated with Hashimoto's thyroiditis. PMID- 19809804 TI - Thyroid hormone and cortisol concentrations after congenital heart surgery in infants younger than 3 months of age. AB - PURPOSE: To investigate associations between thyroid hormone and cortisol concentrations after cardiac surgery and postoperative intensive care course. METHODS: Prospective observational study of 52 infants <3 months of age following congenital heart surgery. RESULTS: Infants with low tri-iodothyronine (< or =3.7 pmol/l) or high cortisol concentration on postoperative admission to the paediatric intensive care unit (PICU) were more likely to have a high inotrope requirement on postoperative day 2 (p = 0.003 and p = 0.0006, respectively) or need mechanical ventilation for longer than 96 h (p = 0.04 and p = 0.002, respectively). Total cortisol concentration < 200 nmol/l was found in 16 (31%) infants. There was no association between low cortisol and postoperative complications. CONCLUSIONS: Infants <3 months of age with low tri-iodothyronine or high cortisol concentration on PICU admission have a more complicated PICU course. Low cortisol concentration is common in the early postoperative period, but is not associated with postoperative complications. PMID- 19809805 TI - Primary anterior shoulder dislocation in young athletes: fix them! PMID- 19809806 TI - Effect of a biplanar osteotomy on primary stability following high tibial osteotomy: a biomechanical cadaver study. AB - Open-wedge high tibial osteotomy (HTO) is becoming increasingly popular for the treatment of varus gonarthrosis in the active patient. The various implants used in HTO differ with regard to its design, the fixation stability and osteotomy technique. It is assumed that the combination of a plate fixator with a biplanar, v-shaped osteotomy supports bone healing. So far, there are no biomechanical studies that quantify the stabilizing effect of a biplanar versus uniplanar osteotomy. We hypothesized that a significant increase in primary stability of bone-implant constructs is achieved when using a biplanar as opposed to a uniplanar osteotomy. Twenty-four fresh-frozen human tibiae were mounted in a metal cylinder, and open-wedge osteotomy (12 mm wedge size) was performed in a standardized fashion. Proximal and distal tibial segments were marked with tantalum markers of 0.8 mm diameter. Two different plates with locking screws were used for fixation: a short spacer plate (group 1, n = 12) and a plate fixator (group 2, n = 12). In six specimens of each group, a biplanar V-shaped osteotomy with a 110 degrees angulated anterior cut behind the tuberosity parallel to the ventral tibial shaft axis was performed. In the remaining six specimens of each group, a simple uniplanar osteotomy was performed in an oblique fashion. Axial compression of the tibiae was performed using a material testing machine under standardized alignment of the loading axis. Load-controlled cyclical staircase loading tests were performed. The specimens were radiographed simultaneously in two planes together with a biplanar calibration cage in front of a film plane with and without load after each subcycle. Radiostereometry allowed for serial quantification of plastic and elastic micromotion at the osteotomy site reflecting the stability provided by the combination of implant and osteotomy technique. No significant additional stabilizing effect of a biplanar osteotomy in craniocaudal and mediolateral plane was found. However, additional stability was achieved in anteroposterior (AP) and all rotational planes in those specimens fixated with a short spacer plate. In this biomechanical set-up with axial load, the additional stabilizing effect of a biplanar osteotomy did not come into effect in the presence of a long and rigid plate fixator. However, biplanar osteotomy increased the fixation stability significantly in AP and rotational planes when a short spacer plate was used. Clinically, the biplanar osteotomy promotes bone healing regardless of the implant used. Biomechanically, biplanar osteotomy is advantageous for shorter plate designs to increase primary stability of the bone-implant construct. PMID- 19809807 TI - Proteolytic processing of Escherichia coli twin-arginine signal peptides by LepB. AB - The twin-arginine translocation (Tat) apparatus is a protein targeting system found in the cytoplasmic membranes of many prokaryotes. Substrate proteins of the Tat pathway are synthesised with signal peptides bearing SRRxFLK 'twin-arginine' amino acid motifs. All Tat signal peptides have a common tripartite structure comprising a polar N-terminal region, followed by a hydrophobic region of variable length and a polar C-terminal region. In Escherichia coli, Tat signal peptides are proteolytically cleaved after translocation. The signal peptide C terminal regions contain conserved AxA motifs, which are possible recognition sequences for leader peptidase I (LepB). In this work, the role of LepB in Tat signal peptide processing was addressed directly. Deliberate repression of lepB expression prevented processing of all Tat substrates tested, including SufI, AmiC, and a TorA-23K reporter protein. In addition, electron microscopy revealed gross defects in cell architecture and membrane integrity following depletion of cellular LepB protein levels. PMID- 19809808 TI - Repeated amphetamine administration in rats revealed consistency across days and a complete dissociation between locomotor and hypothalamic-pituitary-adrenal axis effects of the drug. AB - RATIONALE: Most drugs of abuse stimulate both locomotor activity and the hypothalamic-pituitary-adrenal (HPA) axis, but the relationship between the two responses within the same subjects and their reliabilities has been scarcely studied. Our objectives were to study: (1) the consistency and stability across time of locomotor and HPA activation induced by repeated d-amphetamine (AMPH); (2) the relationship between locomotor and hormonal responses to AMPH; and (3) the relationship between novelty-induced activity and both types of responses to the drug. METHODS: Male adult rats were exposed to a novel environment to study the locomotor response. Later, they were injected with AMPH (2 mg/kg, sc) for 5 days. In Experiment 1, Plasma adrenocorticotropin (ACTH) and corticosterone levels in response to AMPH were studied on days 1, 3, and 5, and locomotor response on days 2 and 4. In Experiment 2, ACTH and corticosterone responses were studied on days 2 and 4, and locomotor response on days 1, 3, and 5. RESULTS: Across days, both locomotor and HPA responses to the drug were consistent, but independent measures, unrelated to the reactivity to novelty. As measured by the area under the curve, the HPA response to AMPH desensitized with the repeated injection, whereas the initial locomotor response to the drug increased. CONCLUSIONS: Dissociation exists between HPA and locomotor activation induced by AMPH, which seemed to be both reliable individual traits. Locomotor reactivity to novelty was related neither to HPA nor to locomotor responses to AMPH. PMID- 19809809 TI - A fenugreek seed extract selectively reduces spontaneous fat consumption in healthy volunteers. AB - PURPOSE: Fenugreek seeds (Trigonella foenum-graecum L.) are an old herbal remedy used to treat metabolic and nutritive dysfunctions. They have been shown to modulate feeding behaviour in animals, but strong clinical data are lacking. The aim of this study was to investigate the effects of a repeated administration of a fenugreek seed extract on energy intake and eating behaviour in healthy human volunteers. METHODS: Twelve healthy male volunteers completed a double-blind randomized placebo-controlled three-period cross-over trial of two different doses of a fenugreek seed extract (588 and 1176 mg). The three 14-day treatment periods were separated by a 14-day washout period. The main endpoints were energy intake, assessed in volunteers under normal ambulatory and free-living conditions by a 3-day detailed dietary record and during a meal test, weight, fasting glucose level, insulin and lipid profile, visual analogue scale scores of appetite/satiety and blood glucose and insulin levels measured repeatedly after a standardized breakfast. RESULTS: Daily fat consumption was significantly decreased by the higher dose of fenugreek seed extract [3.73 vs. 4.51 MJ day(-1), -17.3% vs. placebo, 95% confidence interval (CI) -1.51 to -0.05, n = 12, P = 0.038]. This specific reduction tended to lower the total energy intake (9.97 vs. 11.29 MJ day(-1), -11.7% vs. placebo, 95% CI -2.91 to 0.26, n = 12, P = 0.094). No significant effect was observed on the other nutrients or other endpoints. CONCLUSIONS: The repeated administration of a fenugreek seed extract specifically decreases dietary fat consumption in humans which, given the traditional use of the plant, constitutes a novel result. PMID- 19809811 TI - ATP-citrate lyase activity and carotenoid production in batch cultures of Phaffia rhodozyma under nitrogen-limited and nonlimited conditions. AB - ATP-citrate lyase (ACL) is the key cytoplasmic enzyme which supplies acetyl-CoA for fatty acids in oleaginous yeast. Although it has been suggested that fatty acid and carotenoid biosynthesis may have a common source of acetyl-CoA in Phaffia rhodozyma, the source for carotenoids is currently unknown. The purpose of this work was to analyze the development of ACL activity during batch cultures of P. rhodozyma under ammonium-limited and nonammonium-limited conditions and study its possible relationship with carotenoid synthesis. Every experiment showed carotenoid accumulation linked to an increasing ACL activity. Moreover, the ACL activity increased with dissolved oxygen (DO), i.e., ACL responded to DO in a similar way as carotenoid synthesis. Additionally, in the ammonium-limited culture, ACL activity increased upon ammonium depletion. However, the contribution to carotenoid accumulation in that case was negligible. This suggests that P. rhodozyma has developed two components of ACL, each one responsive to a different environmental stimulus, i.e., DO and ammonium depletion. The role of each component is still unknown; however, considering that the former responds to DO and the known role of carotenoids as antioxidants, it may be a provider of acetyl-CoA for carotenoid synthesis. PMID- 19809812 TI - Enhanced biological denitrification of high concentration of nitrite with supplementary carbon source. AB - Nitrite accumulates during biological denitrification processes when carbon sources are insufficient. Acetate, methanol, and ethanol were investigated as supplementary carbon sources in the nitrite denitrification process using biogranules. Without supplementary external electron donors (control), the biogranules degraded 200 mg l(-1) nitrite at a rate of 0.27 mg NO(2)-N g(-1) VSS h(-1). Notably, 1,500 mg l(-1) acetate and 700 mg l(-1) methanol or ethanol enhanced denitrification rates for 200 mg l(-1) nitrite at 2.07, 1.20, and 1.60 mg NO(2)-N g(-1) VSS h(-1), respectively; these rates were significantly higher than that of the control. The sodium dodecyl sulfate polyacrylamide gel electrophoresis of the nitrite reductase (NiR) enzyme identified three prominent bands with molecular weights of 37-41 kDa. A linear correlation existed between incremental denitrification rates and incremental activity of the NiR enzyme. The NiR enzyme activity was enhanced by the supplementary carbon sources, thereby increasing the nitrite denitrification rate. The capacity of supplementary carbon source on enhancing NiR enzyme activity follows: methanol > acetate > ethanol on molar basis or acetate > ethanol > methanol on an added weight basis. PMID- 19809810 TI - HTR1B as a risk profile maker in psychiatric disorders: a review through motivation and memory. AB - PURPOSE: Serotonin receptor 1B (HTR1B) is involved in the regulation of the serotonin system, playing different roles in specific areas of the brain. We review the characteristics of the gene coding for HTR1B, its product and the functional role of HTR1B in the neural networks involved in motivation and memory; the central role played by HTR1B in these functions is thoroughly depicted and show HTR1B to be a candidate modulator of the mnemonic and motivationally related symptoms in psychiatric illnesses. METHODS: In order to challenge this assessment, we analyze how and how much the genetic variations located in the gene that codes for HTR1B impacts on the psychiatric phenotypes by reviewing the literature on this topic. RESULTS: We gathered partial evidence arising from genetic association studies, which suggests that HTR1B plays a relevant role in substance-related and obsessive compulsive disorders. On the other hand, no solid evidence for other psychiatric disorders was found. This finding is quite striking because of the heavy impairment of motivation and of mnemonic-related functions (for example, recall bias) that characterize major psychiatric disorders. CONCLUSIONS: The possible reasons for the contrast between the prime relevance of HTR1B in regulating memory and motivation and the limited evidence brought by genetic association studies in humans are discussed, and some suggestions for possible future directions are provided. PMID- 19809813 TI - Repair of fresh patellar tendon rupture: tension regulation at the suture line. AB - The reported complications of the repaired patellar tendon have been attributed to the influence of the mechanical environment on the healing process. This study postulates that the healing complications can be minimised through tension regulation at the suture line using an absorbable reinforcement device. Twelve patients with fresh patellar tendon rupture were included in the study. They were prospectively followed up for an average period of 45 months. The patients resumed their pre-injury activities at an average of 6.1 months. The active knee movement averaged 0-154.6 degrees compared to 0-156.7 degrees in the contralateral knee. Radiologically no patella alta, patella baja or degenerative changes in the patellofemoral joints were noted. The results support use of the absorbable reinforcement device for tension regulation at the suture line. PMID- 19809814 TI - Relationship of mRNA expressions of RanBP2 and topoisomerase II isoforms to cytotoxicity of amrubicin in human lung cancer cell lines. AB - PURPOSE: RanBP2 is a small ubiquitin-like modifier ligase for DNA topoisomerase II (TopoII) and plays a role in maintaining chromosome stability by recruiting TopoII to centromeres during mitosis. Engineered-mice with low amounts of RanBP2 have been reported to form lung adenocarcinomas. Furthermore, in the murine embryonic fibroblasts, formation of chromatin bridges in anaphase, a distinctive feature of cells with impaired DNA decatenation by chemical inhibition of TopoII, has been reported. In this study, we tested whether the association between mRNA expression of the RanBP2 gene and chemosensitivity of a TopoII inhibitor, amrubicin could be seen. METHODS: Using a panel of 20 lung cancer cell lines, the mRNA expression levels of the RanBP2, TopoII-alpha and TopoII-beta genes were examined by quantitative real-time reverse transcription PCR. The in vitro cytotoxicity of amrubicin was assessed using a tetrazolium-based colorimetric assay (MTT assay). RESULTS: Although RanBP2 mRNA expression was infrequently downregulated in human lung cancer cell lines, significantly higher RanBP2 transcripts were observed in small cell lung cancer than non-small cell lung cancer. There were no correlations between chemosensitivity of amrubicin and mRNA expression levels of the RanBP2, TopoII-alpha and TopoII-beta genes. CONCLUSIONS: Our in vitro results suggest that mRNA expressions of RanBP2 and TopoII isoforms are unlikely to be a predictive biomarker for the sensitivity to amrubicin. PMID- 19809815 TI - Feasibility study for biweekly administration of cisplatin plus gemcitabine as adjuvant-chemotherapy for completely resected non-small cell lung cancer. AB - PURPOSE: To evaluate the feasibility of biweekly administration of cisplatin and gemcitabine as adjuvant chemotherapy for patients with completely resected non small cell lung cancer (NSCLC). PATIENTS AND METHODS: This was a single-arm, single-institutional study. Patients with completely resected NSCLC (p-Stages IB IIIA) with no previous chemotherapy or radiotherapy were eligible. Simon's optimal two-stage design was applied. Both cisplatin (50 mg/m(2)) and gemcitabine (1,000 mg/m(2)) were given on days 1 and 15, every 28 days. The primary endpoint of this study was the feasibility of this combination in the four cycles of treatment. RESULTS: Twenty patients (19 lobectomies and 1 pneumonectomy) were enrolled in this study. Nine (45%) of patients had grade 3/4 neutropenia, and 6 (30%) had grade 3/4 anemia. Severe non-hematologic toxicities were uncommon in this series. No treatment-related death was encountered. Thirteen (65%) patients completed the planned 4 cycles of chemotherapy. The median intensity was 24 (range 21-25) mg/(m(2) week) with an average of 24.0 (21-25) mg/(m(2) week) cisplatin and 483 (range 412-500) mg/(m(2) week) with an average of 481.0 (412 500) mg/(m(2) week) gemcitabine. The median relative dose intensity of cisplatin was 100 (range 25-100) % with an average of 87.4 (25-100) % and that of gemcitabine was 100 (range 25-100) % with an average of 86.8 (25-100) %. CONCLUSION: This regimen is feasible in the treatment of patients with completely resected NSCLC. A multicenter phase III trial is warranted to assess the efficacy of this regimen at promoting survival and preventing recurrence. PMID- 19809816 TI - Correlation of ESR, C3, C4, anti-DNA and lupus activity based on British Isles Lupus Assessment Group Index in patients of rheumatology clinic. AB - This study aimed to determine the correlation between of ESR, C3, C4, anti-DNA, and lupus activity and also the construct and criterion validity of the British Isles Lupus Assessment Group (BILAG) index for assessing disease activity in systemic lupus erythematosus (SLE). Patients with SLE were recruited into a cross sectional study. Data were analyzed for estimating of SLE disease activity [scores on the BILAG index and Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI-2K)]. Overall BILAG scores were determined by the highest score achieved in any of the individual systems in the respective index. Erythrocyte sedimentation rates (ESRs), C3 levels, C4 levels, anti-double stranded DNA (anti-dsDNA) levels, and SLEDAI-2K scores were used in the analysis of construct validity. Statistical analyses were performed using ordinal logistic regression for construct validity. Of the 100 patients with SLE, 90% were women. Their mean +/- SD age was 31.1 +/- 9.8 years. Increasing overall scores on the BILAG index were associated with increasing ESRs, decreasing C3 levels, decreasing C4 levels, elevated anti-dsDNA levels, and increasing SLEDAI-2K scores (all P < 0.01). These findings show that the ESR, C3, C4, and anti-DNA should be used in the evaluation and management of patients with SLE. Also the results show that the BILAG index has construct validity. PMID- 19809817 TI - Evidences of rheumatic disorders and orthopedic practices in Moche art. PMID- 19809818 TI - Investigation of occurrence of osteonecrosis of the femoral head after increasing corticosteroids in patients with recurring systemic lupus erythematosus. AB - Osteonecrosis (ON) of the femoral head is known to occur commonly in cases with systemic lupus erythematosus (SLE) that received corticosteroid (CS) treatment. However, there have been no detailed reports about the onset of ON in cases with recurrence of SLE. Using MRI, we followed up 17 patients who experienced recurrence of SLE for at least 1 year at our hospital and in whom the CS dose was increased from a maintenance dose to middle to high dose to see if ON would occur. We then compared the group that developed ON and the group that did not with respect to patient characteristics, blood test results, changes in serum lipid levels, and CS dose. ON occurred in five subjects (29.4%), revealing that osteonecrosis occurs not only when CS are first administered but also in cases which the CS dose is increased for recurrence of SLE. Especially, serum cholesterol levels and its rate of increase soared rapidly soon after increasing the CS dose in the ON group as compared with the non-ON group (P < 0.05). This suggests that increased serum lipid levels might be a contributing factor to onset of ON. Moreover, SLE disease activity index 2000 (SLEDAI-2K) scores when the CS dose was increased were significantly (P < 0.05) higher in the ON group, suggesting that SLE disease activity itself is a risk factor for onset of ON. PMID- 19809819 TI - A case of adult-onset Still's disease complicated with atypical pulmonary defect. AB - Adult-onset Still's disease (AOSD) is an uncommon inflammatory condition of unknown origin and pathogenesis. Pulmonary involvement is rare and includes pleural effusion and transient pulmonary infiltrates. We describe and discuss difficulties in the diagnosis of severe respiratory symptoms with uncommon radiological and histological findings and serious prognostic implication in a 24 year-old woman with a severe form of AOSD with destructive arthritis and hematological complications. PMID- 19809820 TI - Monitoring of therapeutic efficacy in a patient with RS3PE syndrome by serologic variables and radiographic methods. AB - We describe a typical case of a patient with remitting seronegative symmetrical synovitis and pitting edema (RS(3)PE) syndrome. He underwent a successful clinical course monitored by serologic variables and radiographic methods. Serum levels of interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), matrix metalloproteinase-3 and serum amyloid A were remarkably elevated. Accumulation of inflammatory cells into the multiple joints was found by gallium-67 scintigraphy. Multiple and symmetrical tenosynovitis with hypervascularity in the presence of subcutaneous edema of the hands and feet were determined by magnetic resonance imaging (MRI) and ultrasonography. These serologic and radiographic abnormalities immediately improved after treatment with a low-dose steroid. Our present case supports a previous observation that synovial tissue is a major inflammatory source of RS(3)PE syndrome. IL-6 (and VEGF), probably produced from the synovial tissues, are considered to be essential factors in the development of RS(3)PE syndrome. PMID- 19809821 TI - Regulation of TNFalpha and IL1beta in rheumatoid arthritis synovial fibroblasts by leukotriene B4. AB - Arachidonic acid metabolites, such as leukotriene B4 (LTB4), are known to play an important role in pathogenesis of rheumatoid arthritis (RA). Apart from leukocytes, RA synovial fibroblasts (RASF) produce a broad array of inflammatory mediators to recruit, retain and activate immune cells and resident mesenchymal cells in the joints to promote ongoing inflammation and tissue destruction. To determine how LTB4 may contribute to this process, RASF was cultured from synovial tissues collected from RA patients undergoing total knee replacement. The level of LTB4 in culture medium was determined using ELISA, and expression of LTB4 receptors (BLT1 and BLT2) by RT-PCR. In the presence of exogenous LTB4, mRNA and protein levels of tumor necrosis factor-alpha (TNFalpha) and interleukin 1beta (IL1beta) were determined by real-time PCR and ELISA. Furthermore, we examined the effects of leukotrienes synthesis inhibitors, MK886 and bestatin, on mRNA and protein levels of TNFalpha and IL1beta in RASF. We found that LTB4 was present at a low concentration in the culture medium of RASF, and the major LTB4 receptor expressed in RASF was BLT2. LTB4 synthesis was activated by treatment with LIT (LPS, ionomycin and thapsogargin), and suppressed by MK886 and bestatin. Exogenous LTB4 remarkably increased the expression of TNFalpha and IL1beta at both the mRNA level and the protein level. In contrast, MK886 and bestatin significantly inhibited their expression. These data suggested that LTB4 contributed to RA by regulating the expression of TNFalpha and IL1beta in RASF. BLT2 was probably the major receptor mediating such effects. PMID- 19809823 TI - [Tears from a medical historical perspective]. PMID- 19809822 TI - The clinical analysis of young children's urolithiasis due to melamine-tainted infant formula. AB - OBJECTIVE: To analyze etiology, clinical features, effective diagnostic methods and therapeutic efficacy of infant urolithiasis related to melamine. METHODS: A total of 2,235 children fed with milk products were screened at the Second Hospital of Shandong University between 1st September and 31st December 2008. Of the children screened, 1,242 were male and 993 female, with a median age of 15 months (range 4-72 months). Among them, 182 with detailed data were enrolled and divided into Group 1 (calculus group, n = 79) and Group 2 (noncalculus group, n = 103) according to their results of the screening. All cases were reviewed with regard to melamine exposure, presentation, laboratory data, B-ultrasound findings and treatment efficacy. RESULTS: Compared to 103 patients without stones, the 79 patients with stones confirmed by B-ultrasound had significant differences in: melamine daily intake (5.17 +/- 4.53 vs. 2.38 +/- 3.39 mg/kg per day, P < 0.001) and duration of problematic milk feeding (12.53 +/- 8.47 vs. 8.65 +/- 3.40 months, P < 0.001). Most of the patients in Group 1 (65.82%) were asymptomatic. In Group 1, all the patients with kidney stones less than 10 mm (n = 75) were successively treated conservatively, whereas the other four with kidney stones >10 mm required surgical treatment. CONCLUSIONS: Melamine overdose can cause an increased risk of developing kidney stones in children. B-ultrasound is the first choice for the diagnosis of urolithiasis related to melamine. For most patients, the conservative treatment is effective; however, the patients with kidney stones >10 mm, surgery may be needed. PMID- 19809825 TI - Initial experience with laparoscopic Chait Trapdoor cecostomy catheter placement for the management of fecal incontinence in children: outcomes and lessons learned. AB - BACKGROUND: Antegrade enemas administered through a percutaneously placed Chait Trapdoor cecostomy catheter have resulted in a marked improvement in compliance and outcome of patients with fecal incontinence. The percutaneous technique, however, is a two-step procedure that is not performed under direct vision. This report presents the results and lessons learned from our experience with the laparoscopic approach to placement of Chait cecostomy catheters. METHODS: Retrospective review of patients who underwent laparoscopic placement of Chait cecostomy catheters from 1999 to 2008. Data collected included patient demographics, primary diagnosis, hospital stay, complications, follow-up duration and outcome. RESULTS: Seventeen patients, mean age 11.8 + or - 4.2 years (range 5 17), underwent laparoscopic Chait cecostomy catheter placement over a period of 8 years. Median follow-up was 46 + or - 21 months (range 4-67). The primary diagnosis was spina bifida in 82% of patients. There was one intraoperative complication, which consisted of tangential needle placement into the cecum, and required conversion to an open procedure. Mean hospital stay was 3.8 + or - 1.5 days (range 2-7). Emergency department visits related to Chait catheter complications were mainly due to catheter dislodgement and breakage. Long-term complications included accidental dislodgement of the catheter in seven patients (41%), mechanical failure of the catheter (breaks/leaks) in six patients (35%), hypertrophic granulation tissue in six patients (35%), wound infections at the catheter site in three patients (18%), complications related to the use of fasteners in two patients (12%) and ventirculoperitoneal (VP) shunt infection in two patients (11.8%). CONCLUSION: The laparoscopic approach to Chait cecostomy catheter placement is a simple and effective procedure. The rate of long term complications such as catheter dislodgement and mechanical failure, which are responsible for the majority of unplanned ED visits, may be decreased by routine yearly catheter exchanges. VP shunt infections are the most serious complications in this patient population consisting mostly of patients with spina bifida. PMID- 19809827 TI - [Autoinflammation and inflammation. The dark energy in the universe of rheumatology]. PMID- 19809826 TI - Patient's appetite is a good indicator for postoperative feeding: a proposal for individualized postoperative feeding after surgery for colon cancer. AB - BACKGROUND: In modern postoperative management, early and enforced feeding has been implemented. The aim of this study is to determine the feasibility of individualized feeding according to the patient's appetite. METHODS: Patients who underwent elective surgery for colon cancer from January 2007 to December 2008 were studied. Liquid intake was allowed on the day of operation and solid normal meal (1800 Kcal) was served according to the recovery of appetite. RESULTS: Two hundred and nine patients were included. Open and laparoscopic surgeries were performed in 104 and 105 patients, respectively. Solid meal was started by the second postoperative day in 81.3% of patients. Intravenous drip infusion was completed within 1 day of the start of the solid meal in 86.6% of the patients and 182 out of 209 patients (87.1%) did not require drip infusion by the third postoperative day. There were no mortalities and readmission rate was 1.0% (2/209). CONCLUSION: The introduction of feeding according to the recovery of appetite was safe and feasible with no delay in establishing oral intake. PMID- 19809828 TI - Clinical and radiological evaluation of minimally displaced proximal humeral fractures. AB - INTRODUCTION: The purpose of this prospective study was to assess the Constant score and radiographic outcome in 66 patients (mean age 58.7 years/mean follow-up 51 months) with a minimally displaced and/or impacted fracture of the proximal humerus treated with early mobilization. METHOD: Special attention was paid to analyze the specific intrinsic parameters (age, gender, ASA grade and length of physiotherapy), injury-related parameters (classification, osteoporosis) and therapy-related parameters (initial fracture displacement, residual bony deformity after healing, secondary fracture displacement during healing period, non-union, humeral head necrosis and omarthrosis) that may influence the final score. PATIENTS: There were 31 A (47%), 22 B (33%) and 13 C-fractures (19%). The median Constant score for the fractured shoulder was 89 points. RESULTS: All fractures healed without non-union. The radiological assessment showed in 80% a fracture-displacement with <15 degrees angulation and/or <5-mm displacement of the greater tuberosity. At time of follow-up, the residual bony-deformity was perfect and good in 88% of cases. There was a significant association between the final Constant score and the age, ASA classification, AO (ABC) classification and initial fracture displacement. CONCLUSION: Early physiotherapy, with a short period of immobilization is a sufficient therapy for management of minimally displaced and/or impacted fractures of the proximal humerus. PMID- 19809829 TI - Comparison of local metronidazole and a local antiseptic in the treatment of bacterial vaginosis. AB - OBJECTIVE: Bacterial vaginosis (BV) is characterized by a mixed flora of pathogenic anaerobic bacteria and associated with risks of pathologic conditions. In the present study, therapy with a local antiseptic spray (octenidine hydrochloride/phenoxyethanol, OHP) for 7 or 14 days is compared against the standard local therapy of BV (metronidazole) in a Serbian patient population. METHODS: As much as 450 women were treated in groups with either 7 days metronidazole vaginal tablets, 7 days OHP, or 14 days OHP. Control smears were taken after each treatment period. RESULTS: In total, 63.2% of the women were without indications of BV after therapy (metronidazole: 61.0%, OHP 7 days: 57.6%, and OHP 14 days: 71.0%). Significantly fewer women were affected from infections after treatment with 14 days OHP compared to OHP for 7 days. CONCLUSIONS: Octenidine hydrochloride/phenoxyethanol spray was as effective as the standard therapy with metronidazole. Patients stated that OHP was more comfortable, easier to apply, and side effects were lesser. PMID- 19809830 TI - The fluid mechanics of scleral buckling surgery for the repair of retinal detachment. AB - BACKGROUND: Scleral buckling is a common surgical technique used to treat retinal detachments that involves suturing a radial or circumferential silicone element on the sclera. Although this procedure has been performed since the 1960s, and there is a reasonable experimental model of retinal detachment, there is still debate as to how this surgery facilitates the re-attachment of the retina. METHODS: Finite element calculations using the COMSOL Multiphysics system are utilized to explain the influence of the scleral buckle on the flow of sub retinal fluid in a physical model of retinal detachment. RESULTS: We found that, by coupling fluid mechanics with structural mechanics, laminar fluid flow and the Bernoulli effect are necessary for a physically consistent explanation of retinal reattachment. Improved fluid outflow and retinal reattachment are found with low fluid viscosity and rapid eye movements. A simulation of saccadic eye movements was more effective in removing sub-retinal fluid than slower, reading speed, eye movements in removing subretinal fluid. CONCLUSIONS: The results of our simulations allow us to explain the physical principles behind scleral buckling surgery and provide insight that can be utilized clinically. In particular, we find that rapid eye movements facilitate more rapid retinal reattachment. This is contradictory to the conventional wisdom of attempting to minimize eye movements. PMID- 19809831 TI - Organization of membrane motor in outer hair cells: an atomic force microscopic study. AB - Using atomic force microscopy, we imaged the cytosolic surface of the lateral plasma membrane of outer hair cells from guinea pigs' inner ear. We used a "cell free" preparation, in which a patch of plasma membrane was firmly attached to a substrate and the cytoplasmic face was exposed. The membrane patches contained densely packed particles whose diameter, after correcting for the geometry of the probing tip, was approximately 10 nm. The particles were predominantly aligned unidirectionally with spacing of approximately 36 nm. The density of the particle was approximately 850 microm(-2), which could be an underestimate presumably due to the method of sample preparation. Antibody-labeled specimens showed particles more elevated than unlabeled preparation indicative of primary and secondary antibody complexes. The corrected diameters of these particles labeled with anti actin were approximately 12 nm while that with antiprestin were approximately 8 nm. The alignment pattern in antiprestin-labeled specimens resembled that of the unlabeled preparation. Specimens labeled with actin antibodies did not show such alignment. We interpret that the particles observed in the unlabeled membranes correspond to the 10-nm particles reported by electron microscopy and that these particles contain prestin, a member of the SLC26 family, which is essential for electromotility. PMID- 19809832 TI - Histone modifications associated with drought tolerance in the desert plant Zygophyllum dumosum Boiss. AB - Zygophyllum dumosum Boiss. is a perennial Saharo-Arabian phytogeographical element and a dominant shrub on the rocky limestone southeast-facing slopes of the Negev desert. The plant is highly active during the winter, and semideciduous during the dry summer, i.e., it sheds its leaflets, while leaving the thick, fleshy petiole green and rather active during the dry season. Being resistant to extreme perennial drought, Z. dumosum appears to provide an intriguing model plant for studying epigenetic mechanisms associated with drought tolerance in natural habitats. The transition from the wet to the dry season was accompanied by a significant decrease in nuclear size and with posttranslational modifications of histone H3 N-terminal tail. Dimethylation of H3 at lysine 4 (H3K4)--a modification associated with active gene expression--was found to be high during the wet season but gradually diminished on progression to the dry season. Unexpectedly, H3K9 di- and trimethylation as well as H3K27 di- and trimethylation could not be detected in Z. dumosum; H3K9 monomethylation appears to be prominent in Z. dumosum during the wet but not during the dry season. Contrary to Z. dumosum, H3K9 dimethylation was detected in other desert plants, including Artemisia sieberi, Anabasis articulata and Haloxylon scoparium. Taken together, our results demonstrate dynamic genome organization and unique pattern of histone H3 methylation displayed by Z. dumosum, which could have an adaptive value in variable environments of the Negev desert. PMID- 19809833 TI - Prime-boost and recombinant protein vaccination strategies using Sm-p80 protects against Schistosoma mansoni infection in the mouse model to levels previously attainable only by the irradiated cercarial vaccine. AB - Advent of an effective schistosome vaccine would contribute significantly toward reducing the disease spectrum and transmission of schistosomiasis. We have targeted a functionally important antigen, Sm-p80, as a vaccine candidate because of its consistent immunogenicity, protective and antifecundity potentials, and important role in the immune evasion process. In this study, we report that using two vaccination approaches (prime boost and recombinant protein), Sm-p80-based vaccine formulation(s) confer up to 70% reduction in worm burden in mice. Animals immunized with the vaccine exhibited a decrease in egg production by up to 75%. The vaccine elicited strong immune responses that included IgM, IgA, and IgG (IgG1, IgG2a, IgG2b, and IgG3) in vaccinated animals. Splenocytes proliferated in response to Sm-p80 produced Th1 and Th17 response enhancing cytokines. These results again emphasize the potential of Sm-p80 as a viable vaccine candidate for schistosomiasis. PMID- 19809834 TI - Impact of oxygen level in gaseous phase on gene transcription and ganoderic acid biosynthesis in liquid static cultures of Ganoderma lucidum. AB - Liquid static cultivation of Ganoderma lucidum was previously found to be very efficient for improving the production of its valuable antitumor compound ganoderic acid (GA) (Fang and Zhong in Biotechnol Prog 18:51-54, 2002). In this work, effects of oxygen concentration within the range of 21-100% (v/v) in the gaseous phase on the mycelia growth, GA production, and gene transcription of key enzymes for GA biosynthesis in liquid static cultures of G. lucidum were investigated. A high cell density of 29.8 +/- 1.7 g/l DW and total GA production of 1427.2 +/- 74.2 mg/l were obtained under an optimal gaseous O(2) level of 80%. The expression of 3-hydroxy-3-methyl-glutaryl-CoA reductase, squalene synthase and lanosterol synthase genes of GA biosynthetic pathway as detected by quantitative real-time PCR was also affected by the gaseous oxygen concentration in the liquid static culture. H(2)O(2) was generated as reactive oxygen species in response to high oxygen concentrations in the gas phase, and it seemed to be involved in the regulation of GA biosynthesis. The information obtained in this study provided an insight into the role of gaseous O(2) in the GA production and it will be helpful for further enhancing its productivity. PMID- 19809835 TI - Extreme hypernatraemia, breast-feeding and red skin. PMID- 19809837 TI - CoMFA and CoMSIA analyses on 4-oxo-1,4-dihydroquinoline and 4-oxo-1,4-dihydro-1,5 , -1,6- and -1,8-naphthyridine derivatives as selective CB2 receptor agonists. AB - Novel classes of CB2 agonists based on 4-oxo-1,4-dihydroquinoline and 4-oxo-1,4 dihydro-1,5-, -1,6- and -1,8-naphthyridine scaffolds have shown high binding affinity toward CB2 receptor and good selectivity over CB1. A computational study of comparative molecular fields analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) was performed, in order to identify the key structural features impacting their binding affinity. The final CoMSIA model resulted to be the more predictive, showing r(2)ncv=0.84, r(2)cv=0.619, SEE = 0.369, and r(2)pred=0.75. The study provides useful suggestions for the synthesis of new selective analogues with improved affinity. PMID- 19809836 TI - Tumor cellular proteasome inhibition and growth suppression by 8-hydroxyquinoline and clioquinol requires their capabilities to bind copper and transport copper into cells. AB - We have previously reported that when mixed with copper, 8-hydroxyquinoline (8 OHQ) and its analog clioquinol (CQ) inhibited the proteasomal activity and proliferation in cultured human cancer cells. CQ treatment of high-copper containing human tumor xenografts also caused cancer suppression, associated with proteasome inhibition in vivo. However, the nature of the copper dependence of these events has not been elucidated experimentally. In the current study, using chemical probe molecules that mimic the structures of 8-OHQ and CQ, but have no copper-binding capability, we dissected the complex cellular processes elicited by 8-OHQ-Cu and CQ-Cu mixtures and revealed that copper binding to 8-OHQ or CQ is required for transportation of the copper complex into human breast cancer cells and the consequent proteasome-inhibitory, growth-suppressive, and apoptosis inducing activities. In contrast, the non-copper-binding analogs of 8-OHQ or CQ blocked the very first step-copper binding-in this chain of events mediated by 8 OHQ-Cu or CQ-Cu. PMID- 19809838 TI - A comparative study of different approaches in the treatment of orbital trauma: an experience based on 274 cases. AB - PURPOSE: The aim of the study was to compare the aesthetic results, complications and surgical indications in patients treated for orbital trauma by one of three different approaches: subciliary, transconjunctival or transconjunctival with lateral canthotomy. MATERIALS AND METHODS: In this retrospective study, 274 patients (169 men and 105 women aged 16-78 years) who had been treated for orbital trauma without soft tissue lacerations of orbital region and then returned for long-term follow-up (6-48 months) between 2000 and 2007 were evaluated in terms of aesthetics and function for the presence of a visible scar and lower-eyelid malposition (scleral show or ectropion). RESULTS: Of the 274 patients, 50 (18.2%) experienced complications. In the group of 219 patients treated with the subciliary approach, 41 experienced complications, whereas amongst the 32 patients treated with the transconjunctival approach, there was one complication (0.3%) and amongst the 23 patients treated with the transconjunctival approach and canthotomy, there were eight complications (34.8%). CONCLUSIONS: Patients treated for orbital trauma with the transconjunctival approach with canthotomy had a higher rate of lower eyelid malposition. Transconjunctival incision without canthotomy was the most successful surgical approach for the treatment of isolated fracture of the orbital floor; however, when major surgical exposure is necessary, subciliary incision is recommended. PMID- 19809840 TI - Musculoskeletal: what's different in children? Fell on arm; favors arm; know what to look for. PMID- 19809839 TI - Subtle pathology detection with multidetector row coronal and sagittal CT reformations in acute head trauma. AB - We sought to analyze retrospectively the advantages of coronal and sagittal reformations obtained with multidetector row computed tomography (CT) in patients with acute head trauma. Multidetector 16-section CT was performed in 200 patients (110 male and 90 female; age range, 3-87 years; mean age, 45 years) with acute head trauma. Scans were performed sequentially, and axial 5-mm-thick slices were obtained from base of skull to vertex. The source data set was reformatted in coronal and sagittal planes, with 2-mm-thick sections at 2-mm intervals. Images were analyzed retrospectively by two independent, blinded readers. The final diagnosis was determined by clinical follow-up. CT imaging abnormalities were detected in 55 out of 200 patients who were scanned for head trauma. Acute traumatic intracranial abnormality was detected on axial scans in 45 patients. Subtle findings were confirmed on coronal and sagittal CT reformations in ten cases, and these were undetected initially on axial CT. Coronal and sagittal reformations confirmed subtle findings in 18.2% (10/55) of the cases (P = 0.001). Indeterminate neuroimaging findings confirmed by coronal and sagittal CT head reformations include tentorial and interhemispheric fissure subdural hemorrhage, subarachnoid hemorrhage, and inferior frontal and temporal lobe contusions. Coronal and sagittal CT head reformations improve the sensitivity and diagnostic confidence in the clinical setting of acute trauma. Overall, coronal and sagittal reformations improved diagnostic confidence and interobserver agreement over axial images alone for visualization of normal structures and in the diagnosis of acute abnormality. PMID- 19809841 TI - Musculoskeletal: what's different in children? Playing on the couch, fell from couch, favoring left upper extremity. PMID- 19809842 TI - The water-deficit stress- and red-rot-related genes in sugarcane. AB - Sugarcane is an important international commodity as a valuable agricultural crop especially in developing countries. Sequencing was carried out to generate >35,000 expressed sequence tags (ESTs) from healthy as well as red-rot-infected tissue of Indian subtropical variety of sugarcane. Subsequent clustering with existing sugarcane ESTs in public databases identified 4,087 clusters, including 85 clusters that preferentially express upon Colletotrichum falcatum (red-rot) infection, which were previously unreported. Real-time reverse transcription-PCR profiling of selected EST clusters identified several sugarcane clusters that show differential expression in response to biotic and abiotic stress conditions. Twenty-five stress-related clusters showed >2-fold relative expression during water-deficit stress in sugarcane. Similarly, EST clusters could be identified, which exhibit association with red-rot disease when assessed in red-rot susceptible and red-rot-resistant varieties of sugarcane. Such EST clusters are good candidates for in-depth analysis to elucidate stress-responsive pathways in sugarcane and facilitate genetic manipulation to tailor this crop for tolerance to various stresses. PMID- 19809843 TI - Fundus white spots and acquired night blindness due to vitamin A deficiency. AB - To report a successfully treated case of acquired night blindness associated with fundus white spots secondary to vitamin A deficiency. An ocular examination, electrophysiologic testing, as well as visual field and OCT examinations were obtained on a 61-year-old man with vitamin A deficiency who had previously undergone gastric bypass surgery. The patient had a re-evaluation after treatment with high doses of oral vitamin A. The patient was observed to have numerous white spots in the retina of each eye. Best-corrected visual acuity was initially 20/80 in each eye, which improved to 20/40-1 OU after oral vitamin A therapy for 2 months. Full field electroretinogram (ERG) testing, showed non-detectable rod function and a 34 and 41% reduction for 32-Hz flicker and single flash cone responses, respectively, below the lower limits of normal. Both rod and cone functions markedly improved after initiation of vitamin A therapy. Vitamin A deficiency needs to be considered in a patient with white spots of the retina in the presence of poor night vision. PMID- 19809844 TI - The results of electrophysiological tests in unilateral extensive peripapillary myelinated nerve fibers: report of three cases. AB - The purpose of this study is to describe clinical and electrophysiological findings of three patients with unilateral extensive peripapillary myelinated nerve fibers. Three patients with unilateral extensive peripapillary myelinated nerve fibers were examined. A complete ophthalmologic examination including uncorrected and best spectacle corrected visual acuities (UCVA, BSCVA) on Snellen charts, dilated fundoscopy, fundus fluorescein angiography (FFA), optical coherence tomography (OCT), perimetry and electrophysiological tests was performed. ERG results of both eyes were normal in all cases. FFA showed hypofluorescence at the myelinated nerve fibers (MNF) area in the affected eyes. We found a visual field defect resembling arcuat scotom at perimetry of the affected eyes. In OCT, there was hyper-reflectivity in reply to MNF. In all cases, pattern ERG results of the affected eyes showed decreased amplitudes and increased latencies of P50 and N95 waves. P100 amplitudes of PVER and FVER were decreased, and latencies were increased in all affected eyes. Extensive peripapillary myelinated nerve fibers may cause changes in PERG and VER. PMID- 19809845 TI - Immune mechanisms in alcoholic liver disease. AB - Growing evidence indicates that inflammatory reactions play an important role in the pathogenesis of alcoholic liver disease (ALD). The implication of immunity in fueling chronic inflammation in ALD has emerged from clinical and experimental evidence showing the recruitment and the activation of lymphocytes in the inflammatory infiltrates of ALD and has received further support by the recent demonstration of a role of Th17 lymphocytes in alcoholic hepatitis. Nonetheless, the mechanisms by which alcohol triggers adaptive immune responses are still incompletely characterized. Patients with advanced ALD show a high prevalence of circulating IgG and T-lymphocytes towards epitopes derived from protein modification by hydroxyethyl free radicals (HER) and end-products of lipid peroxidation. In both chronic alcohol-fed rats and heavy drinkers the elevation of IgG against lipid peroxidation-derived antigens is associated with an increased production of pro-inflammatory cytokines/chemokines and with the severity of histological signs of liver inflammation. Moreover, CYP2E1-alkylation by HER favors the development of anti-CYP2E1 auto-antibodies in a sub-set of ALD patients. Altogether, these results suggest that allo- and auto-immune reactions triggered by oxidative stress might contribute to fuel chronic hepatic inflammation during the progression of ALD. PMID- 19809847 TI - Fibrinogen-dependent signaling in microvascular erythrocyte function: implications on nitric oxide efflux. AB - Experimental evidence has shown that plasma fibrinogen plays a key role as a major cardiovascular risk factor, acting directly to trigger erythrocyte aggregation in occlusive vascular disease. However, due to the complex and hitherto unclear interaction between fibrinogen and the erythrocyte membrane, no study has yet evaluated the effects of fibrinogen, under physiological range values, on the erythrocyte nitric oxide (NO) mobilization. Taking into consideration the potential NO-derived molecules, we have raised the hypothesis that fibrinogen, under physiological conditions, may act to influence blood flow via erythrocyte NO modulation. In this in vitro study whole-blood samples were harvested from healthy subjects, erythrocyte suspensions were incubated in the absence (control aliquots) and presence of different fibrinogen concentrations and levels of NO, nitrite, nitrate and S-nitroglutathione (GSNO) were determined. Our results showed, when compared with control aliquots, that the presence of fibrinogen modulates the NO mobilization in erythrocytes by (1) decreasing erythrocyte NO efflux levels (P < 0.001); (2) increasing levels of intraerythrocytic NO oxidative metabolites, namely, nitrite (P < 0.0001) and nitrate (P < 0.0001); and (3) enhancing the formation of GSNO (P < 0.001). In conclusion, this study provides new insights into an unknown mechanism by which fibrinogen modulates the erythrocyte capacity to supply NO, the effects of which on inflammation profiles (generally associated with blood hyperviscosity and hyperaggregation) still need to be elucidated. Also, increased erythrocyte GSNO levels may be associated with platelet NO metabolism, its activation status and hypotension, which may be extremely relevant in the clinical setting as biomarkers. PMID- 19809848 TI - Investigation of ternary complexes: DNA-phosphatidylcholine liposomes-Mg2+ by freeze-fracture method and their role in the formation of some cell structures. AB - Long-term investigations of ternary complexes: DNA-zwitterionic liposomes divalent metal cations have revealed many details of their structure; but some questions need additional study. The conditions under which fusion or aggregation of liposomes occurs during such complex formation remain obscure. The DNA structure in the ternary complex is still unclear. In this work, using a freeze fracture method, author demonstrate the thin structure of a complex (early attempts to observe this structure employing other electron microscopic methods, in particular cryo-TEM, have not met with success). After treatment of ternary complexes with nuclease S1, which is able to digest single-stranded DNA, local DNA unwinding in such complexes was confirmed. Author describe how the curvature of liposomes as the main factor may determine the interaction between liposomes and DNA, especially aggregation or fusion of liposomes during ternary complex formation. Therefore, interaction between lipids of membrane vesicles in cell and chromatin DNA can be the first stage of a nuclear envelope and pore complex assembly. PMID- 19809849 TI - "Paraduodenal" pancreatitis: results of surgery on 58 consecutives patients from a single institution. AB - BACKGROUND: Cystic dystrophy of heterotopic pancreas, groove pancreatitis, pancreatic hamartoma of the duodenum, paraduodenal wall cyst, and myoadenomatosis are all terms grouped together, from a pathological viewpoint, as definitions of paraduodenal pancreatitis (PP). The objective of the present study was to investigate the clinical characteristics, therapeutic strategies, and results in 58 patients undergoing pancreatic resection for PP. METHODS: From January 1990 to December 2006 data were prospectively collected on 58 patients who were diagnosed with PP who then underwent pancreaticoduodenectomy. RESULTS: In this patient cohort the median age was 44.7 years; only 4 patients were women, and only 3 had non-drinker and/or non smoker habits. The overall morbidity was 18.9%, and the median hospitalization was 11 days. There were no postoperative deaths. In a median follow-up of 96.3 months, all patients noted a decrease in the pain associated with PP; 35 patients (76%) had complete disappearance of pain, whereas occasional relapsing pain occurred in the remaining 11 (24%). CONCLUSIONS: Patients with PP have clinical characteristics similar to those of chronic pancreatitis. The diagnostic imaging modalities of choice are ultrasonographic endoscopy and magnetic resonance cholangiopancreaticography. Based on our surgical experience, pancreaticoduodenectomy seems to be a reasonable choice of treatment in patients with PP. PMID- 19809850 TI - Malnutrition, inflammation: which parameter is interesting to detect in planned oncologic surgery? PMID- 19809852 TI - Nitric oxide measurement in chronic cough. AB - Chronic cough is a common symptom-based diagnostic challenge for primary care physicians and respiratory specialists. Measurement of exhaled nitric oxide is a convenient, reproducible, and inexpensive point-of-service test. It can simplify the evaluation process of chronic cough by providing information regarding the state of airway inflammation. If exhaled nitric oxide is elevated, this is predictive of a favorable response to inhaled corticosteroids. This has the effect of reducing empirical trials for cough-variant asthma or eosinophilic bronchitis. The inclusion of exhaled nitric oxide in the diagnostic approach to chronic cough should be considered. PMID- 19809853 TI - The cough hypersensitivity syndrome: a novel paradigm for understanding cough. AB - For many years patients with chronic cough have been investigated in an attempt to diagnose the cause of the cough. Here I suggest that the overwhelming majority of patients with chronic cough have a single diagnosis: cough hypersensitivity syndrome. This is demonstrated by the homogeneous nature of the clinical history and investigational results of patients attending cough clinics. The hypersensitivity facet of the syndrome is demonstrated by objective testing with capsaicin and other protussive agents. Within the cough hypersensitivity syndrome there are different phenotypes. Those patients with a predominantly Th2-type immune response will develop eosinophilic inflammation and either cough-variant asthma or eosinophilic bronchitis. Those with predominantly heartburn symptoms will have a phenotype that reflects GERD and cough. However, the similarities between the different phenotypes far outweigh differences in a unifying diagnosis of the cough hypersensitivity syndrome, providing a more rational understanding of chronic cough. PMID- 19809854 TI - Author reply: Management of mild elevation of calcitonin after thyroidectomy. PMID- 19809855 TI - On the added value of baseline FDG-PET in malignant lymphoma. AB - PURPOSE: The added value of baseline positron emission tomography (PET) scans in therapy evaluation in malignant lymphoma is unclear. In guidelines, baseline PET is recommended but not mandatory except in lymphoma types with variable fluoro-D glucose uptake. The aim of the present study was to test the hypothesis that adding baseline PET information decreases false positive readings with posttreatment PET and improves observer agreement. METHODS: Forty-four patients (mean age 56 years, standard deviation 14) with malignant lymphoma were included. Two nuclear medicine physicians retrospectively and independently evaluated the posttreatment PET, 3 weeks later followed by paired reading of baseline and posttreatment PET. For each PET, 22 regions were classified as positive, negative, or equivocal, resulting in an overall PET score of positive, unclear, or negative. In case of discrepancies, consensus was reached. RESULTS: Addition of baseline to posttreatment PET evaluation affected the classification of metabolic response in 34% of malignant lymphoma patients treated with first-line chemotherapy. In one out of seven patients, addition of the baseline PET lead to opposite conclusions (95% confidence interval 4-14). False positivity was reduced by adding the baseline scan information, but the effect on false negativity was similar. In addition, the amount of unclear classifications halved after paired reading. Observer agreement did not improve upon adding the baseline PET data. CONCLUSION: Without any other clinical information, pretreatment PET facilitates changes the interpretation of a posttreatment PET in a third of the patients, resulting in both upgrading and downgrading of the posttreatment situation of a malignant lymphoma patient. If these results are confirmed for PET-computed tomography systems, they favor the addition of baseline PET to the current work up of patients with malignant lymphoma. PMID- 19809858 TI - Is television viewing a suitable marker of sedentary behavior in young people? AB - BACKGROUND: Television (TV) viewing is a highly prevalent sedentary behavior in young people and has played a significant role in the assessment of sedentary behaviors. An important question to be addressed is to what extent TV viewing is a suitable indicator, or marker, of overall levels of sedentary behavior in children and adolescents. This has not yet been attempted in youth, but has already been attempted in Australian adults. PURPOSE: This study was conducted to test whether TV viewing in UK teenagers is a marker of sedentary behavior more broadly and to see if the results mirror those of Australian adults. METHODS: Ecological momentary assessment time-use diaries were completed by 561 boys and 923 girls (mean age 14.67 years) in which weekday and weekend out-of-school time behaviors were recorded every 15 min. RESULTS: TV viewing was negatively associated with other leisure-time sedentary behaviors for both boys and girls for weekdays and weekends. Higher levels of TV viewing were associated with less time in other key sedentary behaviors, such as computer use in boys and motorized transport in girls. CONCLUSIONS: TV viewing appears not to reflect additional time in other sedentary behaviors in British teenagers, in contrast to data from Australian women. Studies of sedentary behavior should encompass as wide a range of behaviors as possible. PMID- 19809857 TI - Do tantalum and titanium cups show similar results in revision hip arthroplasty? AB - Porous surfaces are intended to enhance osteointegration of cementless implants. Tantalum has been introduced in an effort to enhance osseointegration potential of uncemented components. We therefore compared the clinical outcome of acetabular components with two different porous surfaces. We retrospectively reviewed 283 patients (295 hips) who underwent cementless revision hip arthroplasty with either an HA-coated titanium cup (207 patients, 214 hips) or porous tantalum cup (79 patients, 81 hips). The minimum followup was 24 months in both groups (titanium: average 51.8 months, range, 24-98 months; tantalum: average, 35.4 months, range, 24-63 months). The titanium and tantalum groups had a mechanical failure rate (clinical plus radiographic) of 8% and 6%, respectively. In hips with minor bone deficiency (type 1, 2A, 2B using the classification of Paprosky et al.), 6% of titanium cups and 4% of tantalum cups failed. In hips with major bone deficiency (type 2C, 3), 24% of titanium cups and 12% of tantalum cups developed failure. In the major bone deficiency group, the tantalum cups had fewer numbers of lucent zones around the cup. Eighty-two percent of titanium cups that failed did so at 6 months postoperatively or later, whereas 80% of tantalum cups that failed did so in less than 6 months. Radiographically in the major group, tantalum cups yielded better fixation. LEVEL OF EVIDENCE: Level III, retrospective comparative study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19809859 TI - Multilevel correlates of satisfaction with neighborhood availability of fresh fruits and vegetables. AB - BACKGROUND: Little is known about influences on perceptions of neighborhood food environments, despite their relevance for food-shopping behaviors and food choices. PURPOSE: This study examined relationships between multilevel factors (neighborhood structure, independently observed neighborhood food environment, and individual socioeconomic position) and satisfaction with neighborhood availability of fruits and vegetables. METHODS: The multilevel regression analysis drew on data from a community survey of urban adults, in-person audit and mapping of food stores, and the 2000 Census. RESULTS: Satisfaction with neighborhood availability of fruits and vegetables was lower in neighborhoods that were farther from a supermarket and that had proportionately more African American residents. Neighborhood poverty and independently observed neighborhood fruit and vegetable characteristics (variety, prices, and quality) were not associated with satisfaction. Individual education modified relationships between neighborhood availability of smaller food stores (small grocery stores, convenience stores, and liquor stores) and satisfaction. CONCLUSIONS: Individual level and neighborhood-level factors affect perceptions of neighborhood food environments. PMID- 19809860 TI - Transesophageal echocardiography detects an unusual complication of infective endocarditis. PMID- 19809861 TI - Corrections to 'The use of sterilised polyester mosquito net mesh for inguinal hernia repair in Ghana'. PMID- 19809862 TI - The association between high plasma homocysteine levels and lower bone mineral density in Slovak women: the impact of vegetarian diet. AB - BACKGROUND: A long-term vegetarian diet is generally poor in vitamin B group. The lack of vitamin B(12) together with vitamin B(6) and folate deficiency is closely related to homocysteine metabolism. Hyperhomocysteinemia was found to be associated with increased bone turnover markers and increased fracture risk. Thus, hyperhomocysteinemia, vitamin B(12) and folate deficiency may be regarded as novel risk factors for micronutrient deficiency-related osteoporosis. AIM OF THE STUDY: To assess the possible impact of a vegetarian diet on bone mineral density in cohort of Slovak vegetarian women. METHODS: Fasting serum glucose, albumin, calcium, phosphorous and creatinine as well as bone markers, serum vitamin B(12), folate and plasma levels of total homocysteine were assessed in two nutritional groups (vegetarians vs. nonvegetarians) of apparently healthy women (age range 20-70 years). Bone mineral density of the femoral neck, trochanter, total femur and lumbar spine was measured in all subjects. RESULTS: Vegetarians had a significantly lower weight (p < 0.05), higher PTH (p < 0.01) and homocysteine (p < 0.001). Vitamin B(12) was significantly higher in nonvegetarians (p < 0.001). No differences were observed in folate levels. Univariate analysis showed significant association between homocysteine and B(12) (p < 0.01), folate (p < 0.001), creatinine (p < 0.001), total proteins (p < 0.049), age (p < 0.001) and vegetarian food intake (p < 0.001). Vegetarians had a significantly lower TrFBMD (p < 0.05) and ToFBMD (p < 0.05). Age and CTx were significant predictors in all sites of measured BMD and PTH. A strong correlation between homocysteine and FNBMD (r = -0.2009, p < 0.002), TrFBMD (r = -0.1810, p < 0.004) and ToFBMD (r = -0.2225, p < 0.001) was found in all subjects. CONCLUSION: Homocysteine is one of the predictors of bone mineral density, and hyperhomocysteinemia is associated with lower bone mineral density. In healthy adults, homocysteine levels are dependent on age as well as on nutritional habits. Thus, elderly women on a vegetarian diet seem to be at higher risk of osteoporosis development than nonvegetarian women. PMID- 19809863 TI - Elevated serum homocysteine level is not associated with serum C-reactive protein in patients with probable Alzheimer's disease. AB - Elevated plasma homocysteine (Hcy) levels have been associated with Alzheimer's disease (AD) and cognitive impairment. Studies have shown that Hcy may have direct and indirect neurotoxicity effects. The aim of the study was to investigate serum Hcy concentration in patients with probable AD with age-matched controls and to determine whether there was an association between serum Hcy and C-reactive protein concentration in patients with probable AD. We also aimed to determine whether there was an association between serum tHcy concentration and cognitive impairment in patients with probable AD. Serum concentration of total Hcy was determined by the fluorescence polarization immunoassay on the AxSYM system, and serum C-reactive protein (CRP) concentration was determined by means of particle-enhanced immunonephelometry with the use of BN II analyzer. Cognitive impairment was tested by the MMSE score. Body mass index (BMI) was calculated for each subject included in the study. Age, systolic and diastolic blood pressure and BMI did not differ significantly between the two groups. Mean serum tHcy concentration in the control group of subjects was 12.60 mumol/L, while in patients with probable AD the mean serum tHcy concentration was significantly higher than 16.15 mumol/L (p < 0.01). A significant negative association between serum tHcy concentration and cognitive impairment tested by the MMSE score in patients with probable AD was determined (r = -0.61634; p < 0.001). Positive, although not significant correlation between CRP and serum tHcy concentrations in patients with AD, was observed. Increased tHcy concentration in patients with probable AD, and the established negative correlation between serum tHcy concentration and cognitive damage tested by MMSE score in the same group of patients, suggests the possible independent role of Hcy in the pathogenesis of AD and cognitive impairment associated with this disease. PMID- 19809864 TI - The dopamine D(2) receptor partial agonist aplindore improves motor deficits in MPTP-treated common marmosets alone and combined with L-dopa. AB - Dopamine replacement therapy in Parkinson's disease (PD) using L-dopa is invariably associated with a loss of drug efficacy ("wearing off") and the onset of dyskinesia. The use of dopamine receptor partial agonists might improve therapeutic benefit without increased dyskinesia expression but may antagonise the effects of L-dopa. We now examine the effects of the novel high affinity, dopamine D(2) receptor partial agonist, aplindore alone and in combination with L dopa in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated common marmoset. In non-dyskinetic MPTP treated animals, aplindore (0.05-1.0 mg/kg p.o.) produced a dose-dependent reversal of motor disability and an increase in locomotor activity that was maximal at doses of 0.2 mg/kg and above. In animals previously exposed to L: -dopa to induce dyskinesia, escalating and repeated dosing of aplindore (0.05-0.5 mg/kg p.o.) produced a sustained, dose-related improvement in motor disability and an increase in locomotor activity. The effects were maximal at a dose of 0.1 mg/kg and above and not different from those produced by L-dopa (12.5 mg/kg plus carbidopa 12.5 mg/kg p.o.). Aplindore administration also led to dose-dependent expression of dyskinesia but at 0.1 mg/kg, this was significantly less intense than that produced by L-dopa. Administration of aplindore (1.0 mg/kg p.o.) in combination with L-dopa (2.5 mg/kg plus carbidopa 12.5 mg/kg p.o.) did not inhibit the reversal of motor deficits but improved motor disability and increased both locomotor activity and dyskinesia expression equivalent to that produced by L-dopa (12.5 mg/kg plus carbidopa 12.5 mg/kg p.o.). These data suggest that dopamine receptor partial agonists would be effective in the treatment of Parkinson's disease and would not inhibit the beneficial actions of L-dopa. PMID- 19809865 TI - Cell-derived soluble oligomers of human amyloid-beta peptides disturb cellular homeostasis and induce apoptosis in primary hippocampal neurons. AB - The concentrations of soluble beta-amyloid (Abeta) oligomers paralleled with the extent of synaptic loss and severity of cognitive impairment in Alzheimer patients. However, the neurotoxicity of the naturally generated Abeta species remains unknown. This study was designed to examine the effects of naturally generated Abeta oligomers, secreted from amyloid precursor protein-expressing cells, on the homeostasis and viability of primary hippocampal neurons. Our results showed that primary hippocampal neurons incubated with condition media containing cell-secreted soluble Abeta had higher levels of heat-shock protein (HSP)27, HSP60 and HSP70, and lower levels of HSP32 than those of the control neurons. The cell-secreted soluble Abeta caused mitochondria dysfunction in hippocampal neurons as demonstrated by depolarized membrane potential and decreased cytochrome c oxidase activity and ATP levels. The levels of pro apoptotic proteins, Bid, Bax and cytochrome C, were elevated; whereas anti apoptotic Bcl-2 protein was reduced in the soluble Abeta-cultured neurons. Apoptosis was also evident in these soluble Abeta-cultured neurons. These results indicate that naturally secreted Abeta induces neuronal injury/death by activating an apoptotic pathway involving impaired mitochondria function and cellular homeostasis. PMID- 19809866 TI - The transactivating effect of HSV-1 ICP0 is enhanced by its interaction with the PCAF component of histone acetyltransferase. AB - ICP0 is a multifunctional protein that plays diverse roles in herpes simplex virus type 1 (HSV-1) infection. It can promote the lytic replication of HSV-1 and activate a variety of viral or cellular genes when introduced into cells by transfection or infection. However, the exact mechanism of ICP0 action is not fully understood. In the present study, we observed the co-localization of ICP0 and PCAF (P300/CBP-associated factor), a component of histone acetyltransferase (HAT), in the ND10 (nuclear dot 10) nuclear body. We further confirmed the interaction between ICP0 and PCAF via yeast two-hybrid assay, co immunoprecipitation, and histone acetyltransferase assays. Analysis of the functional significance of this interaction suggested that PCAF improved the ability of ICP0 to activate transcription of viral genes. Using chromatin immunoprecipitation (ChIP) assays, we observed ICP0-enhanced histone acetylation levels in both viral and cellular gene promoters. Our study suggests that ICP0 regulates transcription through specific interaction with PCAF. PMID- 19809867 TI - Perceptions of high-risk care and barriers to care among women at risk for hereditary breast and ovarian cancer following genetic counseling in the community setting. AB - Data are limited regarding barriers to care among women, with or at risk for hereditary breast and ovarian cancer (HBOC), following genetic counseling in the community setting. Using a telephone survey, we retrospectively addressed perceptions of post-genetic counseling medical care and barriers to care among 69 at-risk women from the non-academic setting. Of these, all agreed that following cancer screening recommendations was better than not following them; none felt recommendations were too difficult to follow; all believed screening would help keep them healthy; 57% believed screening would prevent cancer. Twenty-five percent noted discomfort with breast imaging; 29% found ovarian cancer screening uncomfortable. Close to a quarter of participants reported difficulty deciding whether or not to undergo risk-reducing mastectomy while 10% noted difficulty deciding for or against bilateral salpingo-oophorectomy. There were no perceived major barriers to care, although 38% felt that screening reminders would be helpful, and 10% needed more help in following through with care. Overall, participants believed that they were benefiting from their post-genetic counseling medical care. This work identified HBOC-related support needs to include: informational resources that promote improved understanding of cancer risk and high-risk management; screening reminder systems; and decision support tools. PMID- 19809868 TI - Billing for medical genetics and genetic counseling services: a national survey. AB - In January 2007 the American Medical Association added a new Current Procedural Terminology(R) (CPT) code, 96040, for "Medical Genetics and Genetic Counseling Services." In order to identify the impact of having this new code and to identify issues with implementation of the code, the National Society of Genetic Counselors (NSGC) CPT(R) Working Group surveyed NSGC members using an internet based survey tool. The majority of respondents (94%) reported being aware of the new code and over half of the respondents (69%) said they were billing for genetic counseling. Approximately 24% of those billing reported using 96040. Many facilities are not using this code and the reported success of billing using 96040 is highly varied. Continued education may be beneficial to encourage reimbursement for 96040 and follow up is needed to assess the ongoing implementation and impact of the new CPT(R) code. PMID- 19809869 TI - Resveratrol supplementation gender independently improves endothelial reactivity and suppresses superoxide production in healthy rats. AB - PURPOSE: Resveratrol, a polyphenolic compound mainly abundant in red wines, has beneficial cardiovascular effects on various pathological conditions. However, at present, the effect of resveratrol on health promotion remains unclear. Therefore, in this study, we assessed whether long-term resveratrol supplementation changes endothelial function, vascular contractility, nitric oxide and superoxide production in healthy male and female rats. METHODS: Wistar rats were treated with resveratrol (50 mg/l) in their drinking water for 3 weeks. We investigated relaxation to acetylcholine (10(-9)-10(-4) M) and contractions to phenylephrine (10(-9)-3 x 10(-4) M) and angiotensin II (10(-10)-10(-5) M) in either endothelium-intact or denuded aortae from control and resveratrol-treated male and female rats. Aortic superoxide production capacity was measured in response to provocation by angiotensin II and NAD(P)H. Plasma nitrite/nitrate levels and superoxide dismutase (SOD) activity were also evaluated. RESULTS: Resveratrol supplementation gender independently increased relaxation to acetylcholine and decreased contractions to phenylephrine and angiotensin II in endothelium-intact aortic rings, but not in endothelium-denuded arteries, from healthy male and female rats. This was associated with increased plasma nitrite/nitrate levels. Furthermore, resveratrol caused a refractoriness to angiotensin II and NAD(P)H-induced provocation in superoxide production. CONCLUSION: Our results suggest that resveratrol supplementation gender independently could improve the capacity of endothelial function and suppression of oxidative stress under physiological conditions. Resveratrol ingestion indicates a potential for cardiovascular health promotion. PMID- 19809870 TI - hycu-hr6, A large homologous region of the Hyphantria cunea nucleopolyhedrovirus genome, as a powerful and versatile enhancer in insect expression systems. AB - We previously identified a large homologous region (hr), hycu-hr6, in the genome of the Hyphantria cunea nucleopolyhedrovirus (HycuNPV) and suggested that hycu hr6 was the largest baculovirus promoter enhancer hr identified so far. In this study, we examined the enhancement activity of hycu-hr6 against two promoters from constitutive baculovirus immediate early genes, the HycuNPV ie1 (hycu-ie1) and the Orgyia pseudotsugata multicapsid (M) NPV ie2 (op-ie2), and against a promoter from the inducible Drosophila heat shock protein 70 gene (hsp70) in five lepidopteran (BmN-4, Ld652Y, Sf9, SpIm, and TN368) and one dipteran (S2) insect cell lines. Comparative analyses of transient expression assays using the firefly luciferase gene (luc) as a reporter showed that hycu-hr6 enhanced the activity of all three promoters in all tested cell lines. Comparison of the enhancement efficiency of hycu-hr6 with that of Autographa californica MNPV (AcMNPV) hr5, an enhancer commonly used to improve the activity of AcMNPV ie1 promoter in many insect expression vectors, established that hycu-hr6 was a more efficient enhancer. These results indicate that hycu-hr6 is a versatile and superior promoter enhancer in several insect cell lines and favor the incorporation of hycu-hr6 into insect expression vectors to maximize promoter activity. PMID- 19809871 TI - RNA interference effectively degrades mRNA and inhibits protein expression of GBV C E2 gene in Huh7 cells. AB - The GB virus C/hepatitis G virus (GBV-C/HGV) is a Flaviviridae member that despite its nonpathogenicity, has become of great interest given that it could inhibit the replication of the human immunodeficiency virus (HIV). Therefore, a better knowledge of the viral protein E2 has become our aim. In this study, a GBV C model cell system (HuhEG) which expressing a fusion protein of the GBV-C E2 protein and enhanced green fluorescent protein (EGFP) stably was established. And the expression of these proteins was silenced effectively by the two E2 gene specific siRNAs and an EGFP gene-specific siRNA. This inhibition is sequence specific and extensive (90%). This HuhEG/specific siRNAs system can provide an approach for investigating the association between GBV-C E2 and HIV replication, which may be of potential value in the development of novel prophylactic or therapeutic agents for HIV infection. PMID- 19809872 TI - An update on swine-origin influenza virus A/H1N1: a review. AB - Influenza viruses cause annual epidemics and occasional pandemics that have claimed the lives of millions. The emergence of new strains will continue to pose challenges to public health and the scientific communities. The recent flu pandemic caused by a swine-origin influenza virus A/H1N1 (S-OIV) presents an opportunity to examine virulence factors, the spread of the infection and to prepare for major influenza outbreaks in the future. The virus contains a novel constellation of gene segments, the nearest known precursors being viruses found in swine and it probably arose through reassortment of two viruses of swine origin. Specific markers for virulence can be evaluated in the viral genome, PB1 F2 is a molecular marker of pathogenicity but is not present in the new S-OIV. While attention was focused on a threat of an avian influenza H5N1 pandemic emerging from Asia, a novel influenza virus of swine origin emerged in North America, and is now spreading worldwide. However, S-OIV demonstrates that even serotypes already encountered in past human pandemics may constitute new pandemic threats. There are concerns that this virus may mutate or reassort with existing influenza viruses giving rise to more transmissible or more pathogenic viruses. The 1918 Spanish flu pandemic virus was relatively mild in its first wave and acquired more virulence when it returned in the winter. Thus preparedness on a global scale against a potential more virulent strain is highly recommended. Most isolates of the new S-OIVs are susceptible to neuraminidase inhibitors, and currently a vaccine against the pandemic strain is being manufactured and will be available this fall. This review summarizes the current information on the new pandemic swine-origin influenza virus A/H1N1. PMID- 19809873 TI - Wired for her face? Male attentional bias for female faces. AB - Under conditions of inattention or deficits in orienting attention, special classes of stimuli (e.g. faces, bodies) are more likely to be perceived than other stimuli. This suggests that biologically salient visual stimuli automatically recruit attention, even when they are task-irrelevant or ignored. Here we report results from a behavioral experiment with female and male subjects and two magnetoencephalography (MEG) experiments with male subjects only, in which we investigated attentional capture with face and hand stimuli. In both the behavioral and MEG experiments, subjects were required to count the number of gender-specific targets from either face or hand categories within a block of stimuli. In the behavioral experiment, we found that male subjects were significantly more accurate in response to female than male face target blocks. There was no corresponding effect found in response to hand target blocks. Female subjects did not show a gender-based difference in response to face or hand target blocks. MEG results indicated that the male subjects' responses to face stimuli in primary visual cortex (V1) and the face-selective part of the fusiform gyrus (FG) were reduced when male face stimuli were not relevant to the task, whereas female faces maintained a strong response in these areas in both task relevant and task-irrelevant conditions. These results suggest that within the male brain, female face stimuli are more resilient to suppression than male faces, once attention is drawn to the part of the visual field where the face appears. PMID- 19809874 TI - Genetic diversity assessment of Trollius accessions in China by RAPD markers. PMID- 19809875 TI - Polychlorinated biphenyls, dioxins, furans, and organochlorine pesticides in spotted sandpiper eggs from the upper Hudson River basin, New York. AB - In 2004, spotted sandpipers (Actitis macularia) were studied on the Hudson River near Fort Edward south to New Baltimore, NY and on two river drainages that flow into the Hudson River. Concentrations of 28 organochlorine pesticides, 160 polychlorinated biphenyl (PCB) congeners, and 17 dioxin and furan (PCDD-F) congeners were quantified in eggs collected on and off the Hudson River. The pattern of organochlorine pesticides and PCDD-F congeners did not differ significantly between eggs collected on and off the Hudson River. In contrast, the pattern of PCB congeners differed significantly between the Hudson River and other rivers. Total PCBs were significantly greater in eggs from the Hudson River (geometric mean = 9.1 microg PCBs/g wet weight) than from the other two rivers (0.6 and 0.6 microg PCBs/g wet weight). Seven of 35 (20%) eggs exceeded 20 microg PCBs/g wet weight, the estimated threshold for reduced hatching in tree swallows (Tachycineta bicolor) and some raptor species; the maximum concentration was 72.3 microg PCBs/g wet weight. Models that predicted nest survival and egg success (the proportion of eggs hatching in a clutch if at least one egg hatched) as functions of contaminant levels were poorly distinguished from models that presumed no such associations. While small sample size could have contributed to the inability to distinguish among contaminant and no toxicant models, we cannot rule out the possibility that contaminant concentrations on the Hudson River were not sufficiently high to demonstrate a relationship between contaminant concentrations and reproductive success. PMID- 19809876 TI - Clinical pharmacist interventions on an assertive community treatment team. AB - Assertive Community Treatment (ACT) is a community-based treatment approach intended to help in the recovery and rehabilitation of clients with severe and persistent mental illnesses. A clinical pharmacist is not routinely a member of an ACT team. The purpose of this study is to demonstrate the role of a pharmacist by reviewing recommendations and interventions made by a clinical pharmacist on an ACT team. Information was gathered through a chart review of clients at Community Alternatives in St. Louis, Missouri. All recommendations and interventions performed by the clinical pharmacist between February 1, 2008 and July 31, 2008 were recorded. A total of 341 interventions and recommendations for 29 clients were completed by the pharmacist. Medication management, medication adjustment recommendations, and mental health assessments were the most frequent interventions. This study suggests a clinical pharmacist can be beneficial to an ACT team and provide diverse services to both clients and other team members. PMID- 19809877 TI - The Bax carboxy-terminal hydrophobic helix does not determine organelle-specific targeting but is essential for maintaining Bax in an inactive state and for stable mitochondrial membrane insertion. AB - Here we address the function of the hydrophobic carboxy-terminal tail of the pro apoptotic protein Bax. The tail is tucked into a hydrophobic pocket within the closed/inactive conformation of Bax. Apoptotic stimulation changes the Bax conformation, exposing a mitochondrial-targeting signal. We confirmed that the Bax tail alone can specifically target and anchor a passenger protein to the mitochondria. Surprisingly, we determined that the Bax tail does not play the primary targeting role in Bax mitochondrial translocation. Mutating the Bax tail to produce an ER-targeting signal had no effect on Bax mitochondrial targeting. Additionally, we demonstrated that the Bax tail has a negative regulatory effect on Bax activation. Mutations that disrupt the tail interactions with the hydrophobic pocket resulted in constitutive activation and mitochondrial targeting. Deletion of the Bax tail also resulted in an active conformation of Bax, however, mitochondrial targeting was abolished. Thus, the Bax tail is required for mitochondrial translocation. By generating a mutant-tail that cannot insert into membrane, we determined that insertion of the Bax tail is required for Bax mitochondrial targeting. Our data support a model whereby the Bax tail must be released from the pocket for activation of Bax, then functions as an anchor to stabilize Bax at the mitochondrial membrane after the initial addressing step. PMID- 19809878 TI - Construction of a laser combiner for dual fluorescent single molecule imaging of pRNA of phi29 DNA packaging motor. AB - A customized laser combiner was designed and constructed for dual channel single molecule imaging. The feasibility of a combiner-incorporated imaging system was demonstrated in studies of single molecule FRET. Distance rulers made of dual labeled dsDNA were used to evaluate the system by determining the distance between one FRET pair. The results showed that the system is sensitive enough to distinguish between distances differing by two base pair and the distances calculated from FRET efficiencies are close to those documented in the literature. The single molecule FRET with the dual-color imaging system was also applied to reconstructed phi29 motor pRNA monomers. Finally, techniques for dual laser alignment and tuning of laser power for dual-color excitation are discussed. PMID- 19809880 TI - Reduction of supercritical multiregional stochastic models with fast migration. AB - In this work we study the behavior of a time discrete multiregional stochastic model for a population structured in age classes and spread out in different spatial patches between which individuals can migrate. The dynamics of the population is controlled both by reproduction-survival and by migration. These processes take place at different time scales in the sense of the latter being much faster than the former. We incorporate the effect of demographic stochasticity into the population, which results in both dynamics being modelled by multitype Bienayme-Galton-Watson branching processes. We present a multitype global model that incorporates the effect of both processes and, making use of the existence of different time scales for demography and migration, build a reduced model in which the variables correspond to the total population in each age class. We extend previous results that relate the behavior of the original and the reduced model showing that, given a large enough separation of time scales between demography and migration, we can obtain information about the behavior of the multitype global model through the study of the simpler reduced model. We concentrate on the case where the two systems are supercritical and therefore the expected number of individuals grows to infinity, and show that we can approximate the asymptotic structure of the population vector and the asymptotic population size of the original system through the study of the reduced model. PMID- 19809882 TI - Lead, arsenic, fluoride, and iron contamination of drinking water in the tea garden belt of Darrang district, Assam, India. AB - Drinking water quality with respect to lead, iron, fluoride, and arsenic has been carried out in and around tea gardens of Darrang district of Assam, India. The district lies between 26 degrees 25(') and 26 degrees 55(') northern latitude and 91 degrees 45(') and 91 degrees 20(') east longitude and covers an area of 3,465.30 km(2). Twenty-five different sampling stations were selected for the study. Iron, lead, and arsenic were analyzed by using an atomic absorption spectrometer, Perkin Elmer AA 200, while fluoride was measured by the SPADNS method using a UV-VIS spectrometer, Shimadzu 1240 model. The study revealed that the water sources in the area are heavily polluted with lead. Statistical analysis of the data is presented to determine the distribution pattern, localization of data, and other related information. Statistical observations imply non-uniform distribution of the studied parameters with a long asymmetric tail either on the right or left side of the median. PMID- 19809884 TI - Abstracts and meeting report of the UC Davis Transgenic Animal Research Conference VII, Tahoe City, California, USA, August 17-21, 2009. PMID- 19809881 TI - Genetic variation in N-acetyltransferases 1 and 2, cigarette smoking, and risk of non-Hodgkin lymphoma. AB - Cigarette smoke contains many carcinogens that are metabolically activated through xenobiotic metabolism by phase I and II enzymes, including N acetyltransferases 1 and 2 (NAT1 and NAT2). We investigated non-Hodgkin lymphoma risk in general and by subtype in relation to NAT1 and NAT2 genotypes and cigarette smoking in a population-based case-control study in Connecticut. Of the 535 controls, 53.1% reported ever smoking, and of the 461 cases, 55.7% reported ever smoking. We found a two-fold increased risk of T-cell lymphoma among those possessing the NAT1*10 genotype compared to those with other NAT1 genotypes; including an OR of 2.0 (95% CI: 1.0-2.4) for those heterozygous or homozygous for NAT1*10 genotypes. Rapid acetylator NAT2 phenotype increased the risk of both T cell lymphoma (OR = 3.2; 95% CI: 1.1-9.5) and marginal zone lymphoma (OR = 3.0; 95% CI: 1.0-8.7), though these results were based on a small number of cases. When smoking status and risk of NHL was stratified by NAT1 and NAT2 genotypes, an increased risk of NHL overall was observed in current (OR = 1.7; 95% CI: 1.2-2.4) smokers without the NAT1*10 genotype but not among smokers with the NAT1*10 genotype (p-interaction < 0.01). No association between history of cigarette smoking and risk of NHL overall was observed with any NAT2 genotype. Our results present modest evidence that acetylation rate is associated with risk of NHL for specific subtypes and that the NAT1*10 genotype is an "at-risk" allele. Additionally, our results suggest that the relationship between NHL and smoking status may be modified by common genetic variation in NAT1 but not NAT2. We conclude that these findings require replication in larger studies and ultimately in pooled analyses. PMID- 19809879 TI - Identification and treatment of metabolic complications in pediatric obesity. AB - Metabolic consequences of obesity including insulin resistance, type 2 diabetes mellitus, hyperlipidemia, hypertension, polycystic ovarian syndrome, and non alcoholic fatty liver infiltration are rapidly emerging in the pediatric population. Identifying effective strategies for identifying and treating these obesity related comorbidities in children are crucial to the prevention of future cardiovascular disease and poor health outcomes.This review discusses the pathophysiologic connections between obesity, metabolic disease and cardiovascular risk. Current evidence and recommendations for screening and treatment for the metabolic consequences of pediatric obesity are reviewed. PMID- 19809885 TI - Supplementation of cottonseed, linseed, and noug seed cakes on feed intake, digestibility, body weight, and carcass parameters of Sidama goats. AB - A digestibility, feed intake, and carcass evaluation experiment using 20 yearling intact male Sidama goats weighing 16.4 +/- 0.63 kg (mean +/- SD) was conducted in Ethiopia with the objectives to determine feed intake, digestibility, body weight (BW) gain, and carcass parameters. The treatments included feeding natural pasture hay (T1, control) and supplementation with cottonseed cake (284 g-T2), linseed cake (250 g-T3), and noug seed cake (296 g-T4) on dry matter (DM) basis to supply 85 g crude protein (CP) per head per day. Randomized complete block design for feed intake and BW parameters and complete randomized design for digestibility and carcass parameters were used. Hay DM intake was higher (P < 0.01) for T1 than for the other treatments. T3 promoted higher (P < 0.01) DM (29.3 g/kg W(0.75)/day) and CP (14.1 g/kg W(0.75)/day) intake than T4 (8.9 g/kg W(0.75)/day DM and 4.1 g/kg W(0.75)/day CP). T3 showed better (P < 0.05) organic matter and CP digestibility than T2. Goats in T3 had higher nitrogen intake (P < 0.01) and retention (P < 0.05) than those in T1. Goats in T2 and T3 showed higher (P < 0.05) daily BW gain and final BW than those in T4 and T1. Goats in T2 and T3 had higher (P < 0.05) slaughter weight, empty BW, hot carcass weight, rib-eye muscle area, and dressing percentage on slaughter weight basis than those in T1. The results showed that T2 and T3 had similar effect on CP intake, daily BW gain, and carcass parameters for growing Sidama goats fed natural pasture hay. PMID- 19809886 TI - Ovarian activity in beef and dairy cows with prolonged postpartum period and heifers that fail to conceive. AB - The primary objectives of this study were to investigate incidence of abnormal ovarian cyclicity (AOC) and its type in dairy and beef cows with prolonged postpartum period (>90 days) and in heifers that fail to conceive. A total of 53 animals were included in the study: 17 Friesian crosses, 16 Braford crosses, eight Brangus crosses, and 12 local Kedah-Kelantan (KKX) crosses. These animals were initially checked for absence of pregnancy via palpation per rectum. Blood samples for progesterone analysis were obtained twice a week for 2 to 3 months following their spontaneous oestrous cycle, and all animals were rechecked for pregnancy at the end of the study. Progesterone analysis indicated that 33.9% of the total animals were having AOC: 18.9% with cessation of ovarian cyclicity, 9.4% with prolonged luteal phases (PLP), and 5.7% short luteal phases. The highest incidence was observed in Brangus crosses (62.5%), followed by Braford crosses (43.8%), and Friesian crosses (35.3%). In contrast, no AOC was observed in the local KKX breeds, and all of them were found to be pregnant at the end of the study. A significant difference (p < 0.05) in the incidence of AOC and its type was observed between Kedah-Kelantan crosses and the other breeds. Although not significant (p > 0.05), Friesian crosses showed a higher percentage incidence of AOC than beef cows (40% vs 36.4%), with major types being PLP (26.7%) in dairy and cessation of ovarian cycle (27.3%) in beef cows. Compared with beef heifers, beef cows showed a higher percentage of AOC (36.4% vs 28.6%) where again, cessation of cyclicity was the predominant abnormality. In conclusion, AOC reflected by abnormal endocrine pattern is a possible cause of reduction in fertility for dairy and beef cows beyond 90 days postpartum and heifers that fail to conceive. PMID- 19809887 TI - Bladder cancer incidence and mortality rates compared to ecologic factors among states in America. AB - PURPOSE: Smoking, inadequate vitamin D and pesticide exposure have been linked to bladder cancer (BCa) in past studies. The objective of this study is to explore associations between BCa rates and these risk factors. MATERIALS AND METHODS: BCa incidence and mortality rates among states were compared to smoking; solar ultraviolet (UV) radiation levels and drinking water from a surface water supply (which has greater residual pesticide contamination than groundwater and both are used as sources for drinking water). Lack of health insurance, median family income and urbanization were included to adjust for access to health care and socioeconomic status. RESULTS: BCa incidence and mortality correlated directly with smoking and inversely with solar UV radiation for males and females. BCa mortality correlated directly with drinking surface water for both sexes. Lack of health insurance correlated inversely with BCa incidence for females and trended toward significance for males. Multivariable analyses identified solar UV radiation as the best predictor of BCa incidence in males and solar UV radiation and smoking in females. Solar UV radiation, smoking and drinking surface water were the best predictors of BCa mortality in males, while smoking and drinking surface water were the best predictors of mortality in females. CONCLUSIONS: BCa incidence and mortality for both sexes correlated directly with smoking and inversely with solar UV radiation levels. BCa mortality for both sexes correlated with drinking water from a surface water source. It is hypothesized that BCa mortality risks may increase from drinking water contaminated with low levels of pesticides. PMID- 19809888 TI - Coronary heart disease risk screening: the community pharmacy Healthy Heart Assessment Service. AB - OBJECTIVES: We examined the characteristics and CHD risks of people who accessed the free Healthy Heart Assessment (HHA) service operated by a large UK pharmacy chain from August 2004 to April 2006. METHODS: Associations between participants' gender, age, and socioeconomics were explored in relation to calculated 10-year CHD risks by cross-tabulation of the data. Specific associations were tested by forming contingency tables and using Pearson chi-square (chi(2)). RESULTS: Data from 8,287 records were analysable; 5,377 were at low and 2,910 at moderate-to high CHD risk. The likelihood of moderate-to-high risk for a male versus female participant was significantly higher with a relative risk ratio (RRR) 1.72 (P < 0.001). A higher percentage of those in socioeconomic categories 'constrained by circumstances' (RRR 1.15; P < 0.05) and 'blue collar communities' (RRR 1.13; P < 0.05) were assessed with moderate-to-high risk compared to those in 'prospering suburbs'. CONCLUSIONS: People from 'hard-to-reach' sectors of the population, men and people from less advantaged communities, accessed the HHA service and were more likely to return moderate-to-high CHD risk. Pharmacists prioritized provision of lifestyle information above the sale of a product. Our study supports the notion that pharmacies can serve as suitable environments for the delivery of similar screening services. PMID- 19809889 TI - Characteristics of X- and Y-chromosome specific regions of the amelogenin gene and a PCR-based method for sex identification in red deer (Cervus elaphus). AB - The present study attempts to analyse sequences of the X- and Y-chromosome specific regions of the amelogenin (AMEL) gene in red deer. To this end, primers specific for each form of the gene (AMELX and AMELY) were designed based on bovine genomic sequences and the homologous regions of the genes were sequenced. The obtained sequence of AMELX gene showed high similarity with the corresponding region in cattle (91%) and humans (77%), but this similarity was slightly lower among AMELY genes and showed 87 and 73% of identical nucleotides, respectively. In addition, three single nucleotide polymorphisms (SNPs) were found in the AMELX gene of the female red deer investigated. Comparative analysis of the homologous fragments of the red deer AMELX and AMELY genes confirmed the deletion of an AMELY gene fragment in relation to AMELX. Homology of both sequences was 82% of identical nucleotides in the coding region and 74% in 3' non-coding sequence. The sequences studied showed considerable similarity to homologous fragments of the human and bovine gene, but the structural differences observed lead us to design PCR-based method for sex identification in red deer, based on the presented sequences. PMID- 19809890 TI - Prokaryotic expression and characterization of avian influenza A virus M2 gene as a candidate for universal recombinant vaccine against influenza A subtypes; specially H5N1 and H9N2. AB - The conserved M2 protein of influenza A virus is considered as a promising candidate target for a broad-spectrum, recombinant influenza A vaccine. In the present study, the open reading frame (ORF) of avian influenza A/chicken/Iran/101/1998 (H9N2) M2 gene was amplified then cloned in pAED4, prokaryotic expression vector. M2 protein was produced through the expression of this recombinant expression vector (pAED4-M2) in E. coli BL21 (DE3) strain. The expressed M2 protein was analyzed on SDS-PAGE. Western blot assay was used to examine the immunoreaction of the expressed protein using commercial polyclonal anti-M2 antibody. The antigenicity and biological activity of the recombinant protein was also qualitatively detected on infected MDCK cells surface by immunofluorescence assay using rabbit's immunized antiserum. So, according to the sequence alignment based on the mentioned isolate and the result of immunoassay reaction, it seems recombinant vaccine based on A/chicken/Iran/101/1998(H9N2) M2 protein isolate might cover majority of influenza A virus strains specially H5 and H9 circulating in Iran and neighbor regions significantly. PMID- 19809891 TI - Mineral status and mechanical properties of cancellous bone exposed to hydrogen peroxide for various time periods. AB - Processed cancellous bone has been regarded as one alternative for the treatment of bone defects. In order to avoid immunogenic effects and preserve the natural properties of the bone, the optimal processing method should be determined. To observe the influence of hydrogen peroxide on the mineral status and mechanical properties of cancellous bone for various time periods and find the optimal processing time. Cancellous bone granules from bovine femur condyles were treated with 30% hydrogen dioxide for 0, 12, 24, 36, 48, 60 and 72 h separately. The microstructure and mineral content of the granules were evaluated by ash analysis, Micro-CT, scanning electron micrograph and energy dispersive X-ray. The biomechanical properties were analyzed by applying cranial-caudal compression in a materials testing machine. With increasing exposure to hydrogen peroxide, the BMD and BMC of granules gradually decreased, and the Ca/P molar ratios clearly increased (P < 0.05). Meanwhile, the mineral content of the granules increased from 48.5 +/- 1.3 to 79.5 +/- 2.1%. Substantial decreases in the strength of the granules were observed, and after 48 h severe decreases were noted. The decrease in strength was also evident after normalizing the parameters to the cross sectional area. Granules of bovine cancellous bone matrix should be processed by hydrogen peroxide for 12 to 36 h to fulfill the basic requirements of a bone tissue engineering scaffold. These granules could potentially be useful during orthopedic operations. PMID- 19809892 TI - The effect of old age on apolipoprotein E and its receptors in rat liver. AB - Apolipoprotein E (apoE) is associated with aging and some age-related diseases. The majority of apoE is produced by hepatocytes for the receptor-mediated uptake of lipoproteins. Here, the effects of age on the hepatic expression and distribution of apoE and its receptors were determined using immunofluorescence, Western blots, and quantitative PCR in rat liver tissue and isolated hepatocytes. The expression of apoE mRNA and protein was not influenced significantly by aging. Immunofluorescence studies in isolated hepatocytes showed that apoE was more likely to be co-localized with early endosomes, golgi, and microtubules in isolated old hepatocytes. The mRNA expression of the receptor involved in sequestration of apoE, heparan sulfate proteoglycan was reduced in old age, without any significant effect on the expression of either the low-density lipoprotein receptor or low density-lipoprotein receptor-related protein. Old age is associated with changes in hepatic apoE intracellular trafficking and heparan sulfate proteoglycan expression that might contribute to age-related disease. PMID- 19809893 TI - SPARC: a matricellular regulator of tumorigenesis. AB - Although many clinical studies have found a correlation of SPARC expression with malignant progression and patient survival, the mechanisms for SPARC function in tumorigenesis and metastasis remain elusive. The activity of SPARC is context- and cell-type-dependent, which is highlighted by the fact that SPARC has shown seemingly contradictory effects on tumor progression in both clinical correlative studies and in animal models. The capacity of SPARC to dictate tumorigenic phenotype has been attributed to its effects on the bioavailability and signaling of integrins and growth factors/chemokines. These molecular pathways contribute to many physiological events affecting malignant progression, including extracellular matrix remodeling, angiogenesis, immune modulation and metastasis. Given that SPARC is credited with such varied activities, this review presents a comprehensive account of the divergent effects of SPARC in human cancers and mouse models, as well as a description of the potential mechanisms by which SPARC mediates these effects. We aim to provide insight into how a matricellular protein such as SPARC might generate paradoxical, yet relevant, tumor outcomes in order to unify an apparently incongruent collection of scientific literature. PMID- 19809894 TI - The matricellular functions of small leucine-rich proteoglycans (SLRPs). AB - The small leucine-rich proteoglycans (SLRPs) are biologically active components of the extracellular matrix (ECM), consisting of a protein core with leucine rich repeat (LRR) motifs covalently linked to glycosaminoglycan (GAG) side chains. The diversity in composition resulting from the various combinations of protein cores substituted with one or more GAG chains along with their pericellular localization enables SLRPs to interact with a host of different cell surface receptors, cytokines, growth factors, and other ECM components, leading to modulation of cellular functions. SLRPs are capable of binding to: (i) different types of collagens, thereby regulating fibril assembly, organization, and degradation; (ii) Toll-like receptors (TLRs), complement C1q, and tumor necrosis factor-alpha (TNFalpha), regulating innate immunity and inflammation; (iii) epidermal growth factor receptor (EGF-R), insulin-like growth factor receptor (IGF-IR), and c-Met, influencing cellular proliferation, survival, adhesion, migration, tumor growth and metastasis as well as synthesis of other ECM components; (iv) low-density lipoprotein receptor-related protein (LRP-1) and TGF beta, modulating cytokine activity and fibrogenesis; and (v) growth factors such as bone morphogenic protein (BMP-4) and Wnt-I-induced secreted protein-1 (WISP 1), controlling cell proliferation and differentiation. Thus, the ability of SLRPs, as ECM components, to directly or indirectly regulate cell-matrix crosstalk, resulting in the modulation of various biological processes, aptly qualifies these compounds as matricellular proteins. PMID- 19809897 TI - Risk assessment of food-contact materials: past experience and future challenges. AB - The tiered approaches used in the European Union and the USA for the risk assessment of substances in food-contact materials are based on the principle that the amounts of data required to establish safety-in-use depend on the extent of dietary exposure. Tiered approaches are increasingly recognized as sound approaches for chemical risk assessment, which not only offer consumers reasonable certainty of no harm, but also avoid unnecessary toxicological testing and focus scarce risk-assessment resources on substances of potential concern. Although the European Union and United States approaches were developed separately, their tiered testing recommendations are similar. This paper indicates the savings they have brought in the use of animals and suggests areas where testing recommendations for higher dietary exposures might be further refined. It also discusses some of the future challenges in risk assessment of food-contact materials. PMID- 19809896 TI - Medial tibial stress syndrome: conservative treatment options. AB - Medial tibial stress syndrome (MTSS), commonly known as "shin splints," is a frequent injury of the lower extremity and one of the most common causes of exertional leg pain in athletes (Willems T, Med Sci Sports Exerc 39(2):330-339, 2007; Korkola M, Amendola A, Phys Sportsmed 29(6):35-50, 2001; Hreljac A, Med Sci Sports Exerc 36(5):845-849, 2004). Although often not serious, it can be quite disabling and progress to more serious complications if not treated properly. Often, the cause of MTSS is multi-factorial and involves training errors and various biomechanical abnormalities. Few advances have been made in the treatment of MTSS over the last few decades. Current treatment options are mostly based on expert opinion and clinical experience. The purpose of this article is to review published literature regarding conservative treatment options for MTSS and provide recommendations for sports medicine clinicians for improved treatment and patient outcomes. PMID- 19809895 TI - The topical antimicrobial zinc pyrithione is a heat shock response inducer that causes DNA damage and PARP-dependent energy crisis in human skin cells. AB - The differentiated epidermis of human skin serves as an essential barrier against environmental insults from physical, chemical, and biological sources. Zinc pyrithione (ZnPT) is an FDA-approved microbicidal agent used worldwide in clinical antiseptic products, over-the-counter topical antimicrobials, and cosmetic consumer products including antidandruff shampoos. Here we demonstrate for the first time that cultured primary human skin keratinocytes and melanocytes display an exquisite vulnerability to nanomolar concentrations of ZnPT resulting in pronounced induction of heat shock response gene expression and impaired genomic integrity. In keratinocytes treated with nanomolar concentrations of ZnPT, expression array analysis revealed massive upregulation of genes encoding heat shock proteins (HSPA6, HSPA1A, HSPB5, HMOX1, HSPA1L, and DNAJA1) further confirmed by immunodetection. Moreover, ZnPT treatment induced rapid depletion of cellular ATP levels and formation of poly(ADP-ribose) polymers. Consistent with an involvement of poly(ADP-ribose) polymerase (PARP) in ZnPT-induced energy crisis, ATP depletion could be antagonized by pharmacological inhibition of PARP. This result was independently confirmed using PARP-1 knockout mouse embryonic fibroblasts that were resistant to ATP depletion and cytotoxicity resulting from ZnPT exposure. In keratinocytes and melanocytes, single-cell gel electrophoresis and flow cytometric detection of gamma-H2A.X revealed rapid induction of DNA damage in response to ZnPT detectable before general loss of cell viability occurred through caspase-independent pathways. Combined with earlier experimental evidence that documents penetration of ZnPT through mammalian skin, our findings raise the possibility that this topical antimicrobial may target and compromise keratinocytes and melanocytes in intact human skin. PMID- 19809898 TI - Effect of high-pressure/temperature (HP/T) treatments of in-package food on additive migration from conventional and bio-sourced materials. AB - Migration was assessed during and after two high-pressure/temperature (HP/T) treatments intended for a pasteurization (800 MPa for 5 min, from 20 to 40 degrees C) and a sterilization treatment (800 MPa for 5 min, from 90 to 115 degrees C) and were compared with conventional pasteurization and sterilization, respectively. The specific migration of actual packaging additives used as antioxidants and ultraviolet light absorbers (Irganox 1076, Uvitex OB) was investigated in a number of food-packaging systems combining one synthetic common packaging (LLDPE) and a bio-sourced one (PLA) in contact with the four food simulating liquids defined by European Commission regulations. After standard HP/T processing, migration kinetics was followed during the service life of the packaging material using Fourier transform infrared spectrometer (FTIR) spectroscopy. LLDPE withstood the high-pressure sterilization, whereas it melted during the conventional sterilization. No difference was observed on migration from LLDPE for both treatments. In the case of PLA, migration of Uvitex OB was very low or not detectable for all the cases studied. PMID- 19809899 TI - An issue on pregnancy and childbirth. PMID- 19809900 TI - Determinants for high maternal mortality in multiethnic populations in western China. AB - Our purpose of this study was to investigate determinants and patterns of associations with high maternal mortality in poor and multiethnic populations from the Xinjiang Uigur autonomous region of Western China. The researcher found that the maternal mortality ratio of Xinjiang was very high; almost half of the participants delivered at home without clean delivery, and nearly one-fifth of the participants had not received any medical treatment. Eighty-seven percent of maternal deaths were among ethnic minority groups. In multiethnic areas in Xinjiang, social-culture factors, lack of health resources, and low health services utilization were related to high maternal mortality. PMID- 19809901 TI - Health literacy and its association with the use of information sources and with barriers to information seeking in clinic-based pregnant women. AB - We investigated health literacy and its association with the use of information sources and with barriers to information seeking in clinic-based pregnant women. The Short Test of Functional Health Literacy in Adults (STOFHLA) was used to measure health literacy in 143 English-speaking low-income pregnant women. About 15% of the participants demonstrated low health literacy. Participants with low health literacy were less likely to use the Internet and more likely to have self efficacy barriers than participants with high health literacy. Interventions to promote information-seeking skills and Internet access are indicated for women with low health literacy. PMID- 19809902 TI - Developing standards for postpartum hemorrhage in a resource-limited country. AB - Traditionally, standards of care have been developed by a panel of experts and then implemented by a multidisciplinary team. We studied the feasibility of involving health professionals of all grades and policymakers in the establishment of standards for postpartum hemorrhage (PPH) in Malawi. The team established these standards using evidence from Malawi national guidelines and World Health Organization (WHO) manuals. They agreed on 10 objectives and developed the structure, process, and outcome for each objective. The standards addressed different aspects of prevention, diagnosis, and treatment. The involvement of both health professionals and policymakers might promote ownership, sustainability, and allocation of resources for implementation. PMID- 19809903 TI - The pain that binds us: midwives' experiences of loss and adverse outcomes around the world. AB - Although much has been written about helping women and their families cope with loss related to childbearing, little exists in the literature to guide and support the midwives who witness these losses. We conducted qualitative interviews globally with 22 midwives from nations located on six different continents to begin exploring common themes of experiences and coping methods of midwives involved in adverse perinatal outcomes. The concept of critical incident stress (CIS) is presented as a framework for understanding practitioner reactions that occur after adverse outcomes. Implications for practice, education, and continued research are addressed. PMID- 19809904 TI - The lived experience of women pregnant (including preconception) post in vitro fertilization through the lens of virtual communities. AB - In this study, researchers explore and describe the experience of pregnancy via in vitro fertilization (IVF). The lived experience portrayed herein represents the experience of women from at least seven different countries (the United States, Australia, England, Ireland, Canada, Columbia, and Borneo). Professionals from multiple disciplines may use insights gained from this study to better understand emotional, psychological, and physical health needs of women pregnant post IVF. PMID- 19809906 TI - Two new N-oxide Lycopodium alkaloids from Huperzia serrata. AB - Two new Lycopodium alkaloids, N-oxidehuperzine E (1) and N-oxidehuperzine F (2), along with two known alkaloids, huperzines E (3) and F (4), were isolated from Huperzia serrata (Thunb.) Trev. Their structures were elucidated by spectroscopic and chemical transformations. PMID- 19809907 TI - Essential oil composition of lady's mantle (Alchemilla xanthochlora Rothm.) growing wild in Alpine pastures. AB - The volatile fraction from the aerial part of Alchemilla xanthochlora Rothm. (Rosaceae) was obtained by hydrodistillation, and the chemical composition of the obtained oil was determined by GC/FID and GC/MS. Several compounds were found, belonging to the classes of aldehydes, alcohols, terpenes, esters, acids and hydrocarbons. The major constituents were cis-3-hexenol (11.20+/-0.02%), linalool (10.36+/-0.75%), oct-1-en-3-ol (8.98+/-1.43%), and nonanal (7.83+/-0.93%), followed by myrtenol (4.65+/-0.40%), hexadecanoic acid (4.59+/-0.79%), cis-3 hexenyl acetate (3.92+/-0.15%) and alpha-terpineol (3.10+/-0.26%). PMID- 19809908 TI - Methyl gallate is a natural constituent of maple (Genus Acer) leaves. AB - Methyl gallate was found in ethanolic extracts of red maple (Acer rubrum L.), silver maple (A. saccharinum L.) and sugar maple (A. saccharum Marsh) leaves, but more was present in methanolic extracts. The increased amount of methyl gallate in methanolic extracts was accompanied by a disappearance of m-digallate. It is concluded that only some of the methyl gallate detected in methanolic extracts is an artefact as a result of methanolysis of m-digallate. Its presence in ethanolic extracts is evidence that it is also a natural constituent of maple leaves. PMID- 19809909 TI - A new acylated quercetin glycoside from the leaves of Stevia rebaudiana Bertoni. AB - A new acylated quercetin glycoside quercetin-3-O-(4'''-O-trans-caffeoyl)-alpha-L rhamnopyranosyl-(1-->6)-beta-D-galacopyranoside (1), along with luteolin (2), quercetin (3), luteolin-7-O-beta-D-glucoside (4), apigenin-7-O-beta-D-glucoside (5), quercitrin (6), quercetin-3-O-beta-D-arabinoside (7) and 4,5-di-O-caffeoyl quinic acid (8) have been isolated from the leaves of Stevia rebaudiana Bertoni. The structures of these compounds were determined by spectroscopic methods (1H- and 13C-NMR, IR and MS) and by 2D-NMR experiments. PMID- 19809910 TI - Cytotoxic activity of a flavanone from the stem of Bauhinia variegata Linn. AB - A flavanone has been isolated first time from the stem of Bauhinia variegata, and its structure was identified by colour reactions and spectral analysis. In a search for novel anticancer compounds from medicinal plants, the isolated flavanone was tested for cytotoxic activity against 57 human tumour lines, representing leukaemia, non-small cell lung, colon, central nervous system, melanoma, ovarian, renal, prostate and breast cancers. The results showed that the flavanone has cytotoxic activity against human tumour cell lines. PMID- 19809911 TI - Non-thermal extraction of effective ingredients from Schisandra chinensis Baill and the antioxidant activity of its extract. AB - Schisandra chinensis (Turcz.) Baill has been used as a traditional Chinese medicine in China and Japan. It has been used as an anti-aging drug, a stimulant, a sedative and a tonic drug. Its active components are lignans, including deoxyschisandrin, gamma-schisandrin, schizandrin, gomisin A, and so on. Both deoxyschisandrin and gamma-schisandrin were proved to have the effect of protection against CCl4-induced hepatic damage. The extraction of deoxyschisandrin and gamma-schisandrin from S. chinensis using classical methods, however, are tedious and time consuming. So, ultrahigh pressure extraction (UHPE) was used to extract deoxyschisandrin and gamma-schisandrin from S. chinensis. At the same time, heat reflux and ultrasonic extraction methods were chosen as the comparative methods. UHPE optimum extraction conditions were as follows: 400 MPa pressure, 90% ethanol-water solution, 90:1 liquid:solid ratio, 5 min extraction time; the extraction yields of deoxyschisandrin and gamma-schisandrin were 0.3132 and 0.0612%. The extraction yields of deoxyschisandrin and gamma-schisandrin with heat reflux and ultrasonic extraction methods were 0.280 and 0.0483%, 0.272 and 0.0505%, and the extraction times were 2 h and 20 min, respectively. UHPE had the highest extraction yields with the shortest extraction time of the three extraction methods. Furthermore, the UHPE extract had the highest antioxidant activity. So UHPE can not only shorten the extraction time, but also mostly protect the extracts' bioactivities. In a word, UHPE is an efficient extraction method for the extraction of effective ingredients from S. chinensis. PMID- 19809912 TI - Two new insecticidal sesquiterpene esters from Euonymus japonicus. AB - Two new insecticidal sesquiterpene esters with a beta-dihydroagarofuran sesquiterpene skeleton, ejaponine A (1) and ejaponine B (2), and two known compounds, 3 and 4, were isolated from the EtOAc extract of the root bark of Euonymus japonicus Thunb. by bioassay-guided fractionation. Their chemical structures were elucidated mainly by analysis of MS and NMR spectral data. The LD50 values of ejaponine A (1), ejaponine B (2), 3 and 4 showed against Mythimna separata were 89.2, 98.6, 181.4 and 109.2 microg g(-1), respectively. PMID- 19809913 TI - Phenolic and bis-iridoid glycosides from Strychnos cocculoides. AB - A new bis-iridoid glucoside, cocculoside (1), has been isolated, together with five known compounds from Strychnos cocculoides (Loganiaceae), collected in Tanzania. The structure of cocculoside (1) has been determined as an ester dimer of loganin and secologanin dimethyl acetal between C-7 and C-11. PMID- 19809914 TI - Alkaloids from Piper sarmentosum and Piper nigrum. AB - Detailed chemical studies on the roots of Piper sarmentosum and Piper nigrum have resulted in several alkaloids. The roots of P. sarmentosum gave a new aromatic compound, 1-nitrosoimino-2,4,5-trimethoxybenzene (1). Piper nigrum roots gave pellitorine (2), (E)-1-[3',4'-(methylenedioxy)cinnamoyl]piperidine (3), 2,4 tetradecadienoic acid isobutyl amide (4), piperine (5), sylvamide (6), cepharadione A (7), piperolactam D (8) and paprazine (9). Structural elucidation of these compounds was achieved through NMR and MS techniques. Cytotoxic activity screening of the plant extracts indicated some activity. PMID- 19809915 TI - Purification of aloe polysaccharides by using aqueous two-phase extraction with desalination. AB - A PEG6000/(NH4)2SO4 aqueous two-phase system was chosen as the separation system. The effects of NaCl, Gu(SCN)2 and pH on the partitioning of aloe polysaccharide and bovine serum albumin (BSA) were studied. The result shows that the recovery of aloe polysaccharide was decreased with the increase in pH and the recovery of protein was not changed; NaCl and Gu(SCN)2 can change the partitioning behaviour of the BSA. The separation of BSA and aloe polysaccharides was also carried out. Then, the aloe polysaccharides were obtained by using ultrafiltration membrane. The composition of the polysaccharides was analysed by gas chromatography. The results show that mannose is mainly monosaccharide, and it only contains a few glucose. PMID- 19809916 TI - New cyclopentenones from strain Trichoderma sp. YLF-3. AB - Two new cyclopentenones, trichodermone A (1) and trichodermone B (2), together with a known compound, 3-(3-oxocyclopent-1-enyl) propanoic acid (3), were isolated from the strain Trichoderma sp. YLF-3. Their structures were elucidated on the basis of their spectroscopic data. All compounds were assayed for antibacterial activity. Compound 3 showed activity against Staphyloccocus aureus and Bacillus cereus. PMID- 19809917 TI - Five furostanol saponins from fruits of Tribulus terrestris and their cytotoxic activities. AB - Two new furostanol saponins, terrestroside A, 3-O-{beta-D-xylopyranosyl-(1-->3) [beta-D-xylopyranosyl(1-->2)]-beta-D-glucopyranosyl(1-->4)-[alpha-L rhamnopyranosyl(1-->2)]-beta-D-galactopyranosy}-26-O-beta-D-glucopyranosyl-L-5a furost-20(22)-en-(25R)-3beta,26-diol (1) and terrestroside B, 3-O-{beta-D xylopyran-osyl(1-->3)-[beta-D-xylopyranosyl(1-->2)]-beta-D-glucopyranosyl(1-->4) [alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-galactopyranosy}-26-O-beta-D glucopyranosyl-5a-furostan-12-one-(25R)-22-methoxy-3beta,26-diol (2), together with three known compounds, chloromaloside E (3), terrestrinin B (4) and terrestroneoside A (5) were isolated from the dry fruits of Tribulus terrestris. Furthermore, the inhibitory effects of the compounds on tumour cells were evaluated, and compounds 1-5 showed potential anti-tumour activity. PMID- 19809918 TI - Synthesis of novel acetates of beta-caryophyllene under solvent-free Lewis acid catalysis. AB - Acylation of beta-caryophyllene, a sesquiterpene hydrocarbon with acetic anhydride, was carried out under mild catalytic conditions using Lewis acids such as BF3.Et2O, ZnCl2, FeCl3, I2 and AlCl3 as catalysts. Among these, BF3.Et2O was found to catalyse the reaction most efficiently, yielding three acylated products in 75% yield under solvent-free conditions. The structures were confirmed by NMR and MS techniques. PMID- 19809919 TI - Chemical constituents from the fruits of Hippophae rhamnoides. AB - Ten compounds were isolated from the fruits of Hippophae rhamnoides. On the basis of spectroscopic and chemical methods, the structures of these compounds were elucidated as hippophae cerebroside (1), oleanolic acid (2), ursolic acid (3), 19 alpha-hydroxyursolic acid (4), dulcioic acid (5), 5-hydroxymethyl-2-furancarbox aldehyde (6), cirsiumaldehyde (7), octacosanoic acid (8), palmitic acid (9) and 1 O-hexadecanolenin (10). Among them, 1 was a new compound, and 4-7 and 10 were obtained from the genus for the first time. PMID- 19809921 TI - [Muller-cell-derived neurotrophic factors, a road to neuroprotective therapy in the retina]. PMID- 19809922 TI - [Avastin is not the same as Lucentis, and Lucentis is not the same as Avastin]. PMID- 19809923 TI - [Guidelines of clinical practice of the SERV (Spanish Retina and Vitreous Society): management of ocular complications of diabetes. Diabetic retinopathy and macular oedema]. AB - OBJECTIVE: Diabetes mellitus is considered the most common cause of blindness in the working population of industrialized countries, with diabetic macular edema being the most common cause of decreased visual acuity and proliferative diabetic retinopathy (PDR) being responsible for the most severe visual deficits. We have therefore tried to establish a guide for clinical intervention whose purpose is to provide orientation on the treatment of diabetic retinopathy and its complications. This is necessary at a time when many treatment options have emerged whose role is not yet fully defined. METHOD: A group of expert retina specialists selected by the SERV (Vitreous-Retina Spanish Society) assessed the published results of different treatment options currently available, suggesting lines of action according to the degree of diabetic retinopathy present and the presence or absence of macular edema. RESULTS: PDR is primarily treated with pan retinal photocoagulation. For clinically significant diabetic macular edema without signs of vitreomacular traction, the treatment of choice continues to be focal/grid photocoagulation. Similarly, retinovitreal surgery is indicated for both conditions. The use of antiangiogenic drugs was also analyzed but remains inconclusive. CONCLUSION: Laser therapy is effective in the management of diabetic retinopathy and diabetic macular edema. The role of antiangiogenics is not yet sufficiently defined. PMID- 19809924 TI - [Dry eye and quality of life]. AB - PURPOSE: To evaluate the relation between dry eye severity and quality of life. METHODS: 40 participants (19 with dry eye and 21 normal controls) underwent slit lamp examination and Schirmer test to evaluate dry eye severity. Quality of life was evaluated with OSDI (Ocular Surface Disease Index) and VFQ-25 (Visual Function Questionnaire-25). RESULTS: The OSDI total score was significantly greater in patients suffering dry eye. However, BUT (break-up time) and VFQ-25 total score were significantly lower than in control subjects. In dry eye patients several statistically significant correlations (p<0.05) were found: OSDI with VFQ-25 total score (r=-0.62), BUT with corneal staining (r=-0.50) and Schirmer (0.66), BUT with OSDI total score, OSDI-symptoms and OSDI-triggers (r= 0.56, -0.56, -0.60); corneal staining with OSDI total score and OSDI-symptoms (r=0.55, 0.54), BUT with VFQ-25 total score, VFQ-25 ocular pain, mental function and role function. (r=0.56, 0.51, 0.63, 0.56); corneal staining with VFQ-25 total score, VFQ-25 ocular pain and near vision (r=-0.57, -0.49, -0.62). CONCLUSION: Quality of life is decreased in patients with dry eye. OSDI and VFQ-25 questionnaires are valid instruments for measuring the impact of dry eye disease. BUT and corneal staining provide an indirect measurement of quality of life. PMID- 19809925 TI - [Ophthalmology complaints in Spain]. AB - PURPOSE: We reviewed 90 complaints for allegedly incorrect ophthalmic procedures. Most of these complaints (33%) were related to cataract and refractive surgery (18%). In third position in terms of frequency (14%) were complaints concerning oculoplastic surgery and in fourth position were complaints related to retinal detachment (13%). This was followed by a miscellaneous group, which represented 10% of complaints. About 9% of complaints were related to emergency ophthalmic procedures, while just 3% of complaints were related to glaucoma. METHODS: We analysed within each subgroup the characteristics of the claims; the information given to the patient, those cases in which there existed grounds for considering the ophthalmologist's performance as being incorrect, and court orders that were adopted. RESULTS: The most common groups were cataract and refractive surgery, which together represented more than a half of the complaints. We found in 26% of cases, reports from other doctors criticizing the professional performance a posteriori of an accused ophthalmologist, incorrect or incomplete documents of informed consent, as well as patients asserting that their surgeons made them promises of results, or minimized risks about the proposed operation. CONCLUSIONS: In the great majority of cases, the claim was settled due to a characteristic complication inherent in the surgical technique and present in the document of informed consent signed by the patient. We also observed a minority of cases, particularly in refractive surgery, in which a foreseeable and avoidable complication related to incorrectly prescribed surgical techniques was produced. In these exceptional cases, expert evidence is usually unfavorable and charges are typically laid. It is probable that improved information for patients would reduce the number of these claims. PMID- 19809926 TI - [Intraocular tumor diagnosed two years after perforating trauma]. AB - CASE REPORT: We describe the unusual diagnosis of a ciliary body medulloepithelioma by histopathology of a subretinal membrane obtained from vitreoretinal surgery of a 10-year-old boy. The patient had a history of perforating trauma OS 2 years earlier, and both fundus exam and B-scan ultrasound revealed only a retinal detachment with a subretinal membrane. No detectable mass was present. DISCUSSION: The membrane removed from underneath the peripheral retina revealed a blue cell tumor confirmed by histopathology and immunohistochemistry to be a primitive neuroectodermal tumor. Currently, the patient has been followed for 5 years with no signs of recurrence. PMID- 19809927 TI - [Migraineous anterior optic ischemic neuropathy]. AB - CASE REPORT: We report the case of a 22-year-old man who presented with headache and blurred vision which had started four days previous. A periventricular lesion was found in the magnetic resonance imaging. The patient was diagnosed with demyelinating neuritis and treated with intravenous methylprednisolone. DISCUSSION: After six months visual function had not improved, so the initial diagnosis was probably erroneous. It is likely that the patient suffered from migraineous optic ischemic neuropathy. In this paper we review the scarce literature about this topic, and the role of migraine as a cardiovascular risk factor. PMID- 19809928 TI - [Uveitis masquerade syndrome as a presenting form of diffuse retinoblastoma]. AB - CLINIC REPORT: A 3-year-old boy presented with an intermediate uveitis. Complete ophthalmic exam, ocular ultrasonography, magnetic resonance imaging and computerized tomography of the orbit were inconclusive. Determination of the aqueous humor/serum rate of Lactate dehydrogenase (LDH) was the key for the diagnosis of a diffuse retinoblastoma. DISCUSSION: A masquerade syndrome is the initial presentation in 1-3% of retinoblastomas. Aqueous humor punction is contraindicated in patients with retinoblastoma but it might be the only way to achieve a correct diagnosis in these difficult and very unusual cases: enzymatic assays such as LDH offer a good sensitivity and specificity for the diagnosis of these patients. PMID- 19809929 TI - [Ophthalmology in the poetic work of Jorge Luis Borges (II)]. PMID- 19809930 TI - [The pituitary adenoma of Percy Wyndham Lewis (1882-1957)]. PMID- 19809932 TI - Cerebral palsy in the 21st century: is there anything left to say? AB - In April 2009 I was honoured to be asked to present the Emil Becker Lecture at the 35 (th) annual meeting of the German-Swiss-Austrian Society for Neuropaediatrics (Gesellschaft fur Neuropadiatrie). Focusing on my interest and involvement as a clinician and researcher in cerebral palsy, I took the opportunity to look both back at the past 35 years and forward to what I believe are some of the important clinical challenges that are still to be addressed and solved in the 21 (st) century. This paper is therefore a personal reflection rather than a comprehensive review of the literature in cerebral palsy. It is my hope that these reflections will be of interest to others. PMID- 19809933 TI - Effectiveness of low dose of topiramate following rapid titration in multiply handicapped children and difficult-to-treat epilepsy. AB - This prospective, observational, single arm, monocentric study explored efficacy and tolerability outcomes of rapid oral initiation of topimarate in children with difficult to treat epilepsy. The study population consisted of 19 multiply handicapped children (mean age 4.4 years, range 0.6-15.3 years). The observation period was 12 weeks and included 7 visits. The mean initial dose of topiramate was 1.1 mg/kg body weight/d (range: 0.66-2.67 mg/kg/d) following rapid titration. The mean final dose was 3.3 mg/kg/d (range 0.5-6.7 mg/kg/d). An at least 50% reduction of seizure frequency compared to baseline was observed in 9 of 19 patients (47.4%). Six patients (31.6%) had a slight reduction of seizure frequency (<50%) and 4 patients (21.1%) experienced an increase of seizure frequency. A total number of 29 adverse events were documented in 17 of 19 patients. Most frequently captured were fatigue (26.3% of patients), decreased appetite (15.8%) and psychiatric disturbances (15.8%). No serious adverse events were reported. These data might suggest that in certain clinical circumstances rapid dose escalation with topiramate followed by a low maintenance dose might be a good therapeutic option for pediatric patients with difficult to treat epilepsy. PMID- 19809934 TI - Septic encephalopathy: relationship to serum and cerebrospinal fluid levels of adhesion molecules, lipid peroxides and S-100B protein. AB - Severe septic illness is often associated with cerebral manifestations such was disturbed consciousness and delirium. Little was known about its effect on the CNS. This is the first study in children that has assessed the direct mediators of brain inflammation and injury with sepsis. The serum and CSF concentrations of soluble intracellular adhesion molecule-1 (sICAM-1) (marker of endothelium leukocyte interaction), nitric oxide (NO) and lipid peroxide (LPO) (markers for lipid peroxidation) and S-100B protein (marker of astrocytes activation and injury), were measured in 40 children with sepsis of whom 40% had moderate to severe septic encephalopathy. Serum from 25 normal children was used for comparison. Serum values of sICAM-1, NO, LPO and S100B were elevated in patients compared to controls. The greater elevation of the CSF:serum albumin ratio suggests loss of blood-brain barrier integrity. After normalising for CSF:serum albumin ratio, we demonstrated a significant intrathecal synthesis of NO, LPO and S100B. Patients with encephalopathy had elevated serum and CSF levels of sICAM-1, NO, LPO and S100B compared to sepsis only. This study indicates that the brain is vulnerable in children with sepsis. It also suggests that coordinated interactions between immune system, vascular endothelial cells, CNS barriers, astrocytes and brain lipid peroxides, may contribute to septic encephalopathy. PMID- 19809935 TI - Spontaneous intrauterine "ping-pong" fracture: review and case illustration. AB - We report a case of a closed outer-table parietal "ping-pong" skull fracture occurring in a 4 190-gram female infant born at 39 weeks and 5 days gestation after an uneventful Cesarean section (Apgar scores of 9 and 9 at one and five minutes). There was no maternal history of abdominal trauma during pregnancy and there were no complications or difficulties with Cesarean section delivery. Neurological examination was normal. Computed tomography with three-dimensional reconstruction images showed a 4 x 5 cm depression in the right parietal bone with a medial lucency consistent with a fracture of the superior margin of the skull and leftward deviation of the sagittal suture and sinus. Spontaneous resolution did not occur by one month of age and the skull fracture was repaired with excellent cosmetic results. Rarely has a case of spontaneous intrauterine skull fracture been reported in an atraumatic Cesarean delivery. We believe this fracture resulted from a chronic in utero process without associated trauma as evidenced by deviation of the sagittal suture and sinus. PMID- 19809936 TI - Stroke-like episodes in propionic acidemia caused by central focal metabolic decompensation. AB - Propionic acidemia caused by propionyl-CoA carboxylase deficiency frequently leads to neurologic complications. Herein we report an eleven-year-old patient with propionic acidemia having three stroke-like episodes during a period of 13 months characterized by acute reversible hemiplegia and vegetative symptoms like bradycardia or drowsiness. No biochemical signs of severe metabolic decompensation were detectable in plasma. At all three episodes, EEG was not indicative for status epilepticus, but in the acute episode it showed slowing of background activity emphasized on one side. MRI revealed reversible hyperintensities in cortical grey matter and basal ganglia. During the third episode a lumbar puncture was done in parallel with venous puncture. Concentrations of glutamine (902 micromol/L), glycine (24 micromol/L) and alanine (78 micromol/L) were elevated in CSF. In plasma glycine (1 859 micromol/L) and alanine (608 micromol/L) concentrations were also elevated, whereas the glutamine (458 micromol/L) concentration was normal. CSF/plasma ratios were elevated for glutamine (1.97) and alanine (0.13) and normal for glycine (0.01). We assume that the stroke-like episodes in our patient may be caused by an acute focal cerebral metabolic decompensation, which is detectable by unspecific changes in MRI and by measuring amino acids and lactate in CSF versus plasma. PMID- 19809937 TI - An inherited nonsense R1645X mutation in neuronal sodium channel alpha1-subunit gene in a Turkish patient with severe myoclonic epilepsy of infancy. AB - Severe myoclonic epilepsy of infancy (SMEI) is a well-known catastrophic epileptic syndrome. Several mutations of the sodium channel alpha 1 subunit (SCN1A ) gene were reported in patients with SMEI. Most of the mutations were DE NOVO. Inherited truncating mutations are very rare. Here a patient with a new nonsense mutation (c.4933 C>T; p.R1645X) of the gene is described. This mutation was inherited from the father who had only febrile seizures during childhood. PMID- 19809938 TI - Charcot-Marie-Tooth (CMT) disease 1A with superimposed inflammatory polyneuropathy in children. AB - Charcot-Marie-Tooth (CMT) disease is genetically heterogeneous and subdivided into demyelinating (CMT 1) and axonal (CMT 2) types based on neurophysiology findings. CMT1A, the commonest form associated with duplication of the PMP22 segment on chromosome 17p, often arises in childhood but is generally a slowly progressive disease. We report 2 children presenting with clinical features of an acute inflammatory demyelinating polyneuropathy (AIDP) who were subsequently diagnosed with underlying CMT1A. Both children had neurophysiology and histopathology features consistent with CMT1. Immunoglobulin treatment was initiated considering the evidence of superimposed inflammation and appeared to modify disease progression. Our findings indicate that CMT1A predisposes to a superimposed inflammatory neuropathy. Recognition of this association is difficult, particularly in children without clear family history, but of great importance as immunomodulatory treatment may improve outcome. In addition, we postulate that an underlying genetic polyneuropathy should be suspected if the recovery from AIDP is slower than expected, or incomplete. PMID- 19809939 TI - Increased intracranial pressure caused by non-space-occupying arachnoid cysts: report of two patients. AB - In this article we report on two patients with arachnoid cysts previously treated by shunt implantation presenting with clinical signs of an increased intracranial pressure i. e., papilledema, headache and nausea. Repeated MRI scans showed no alteration of the cerebrospinal fluid circulation and no space-occupying effect of the cysts. Although neuroimaging showed no signs of increased intracranial pressure, neurosurgical exploration was performed and revealed a distinctly increased pressure in both arachnoid cysts. After replacement of the shunt a prompt reduction of papilledema and relief of symptoms was observed. PMID- 19809940 TI - Perfusion abnormalities in hemimegalencephaly. AB - INTRODUCTION: Cerebrovascular changes are rarely discussed in patients with hemimegalencephaly. These alterations have previously been associated with epileptical activity. CASE: We report the case of a 36-week gestation neonate presenting with total right hemimegalencephaly, as demonstrated by a magnetic resonance imaging (MRI) performed in the first days of life. Perfusion-weighted imaging displayed a clear hypervascularization of the right hemisphere. Diffusion tensor imaging showed an arrangement of white matter fibers concentrically around the ventricle on the right hemisphere. AngioMRI showed an obvious asymmetry in the size of the middle cerebral arteries, with the right middle cerebral artery being prominent. The baby was free of clinical seizures during his first week of life. An electroencephalogram at that time displayed an asymmetric background activity, but no electrical seizures. CONCLUSION: Perfusion anomalies in hemimegalencephaly may not necessarily be related to epileptical activity, but may be related to vessel alterations. PMID- 19809941 TI - Startle epilepsy associated with infantile hemiplegia (SEIH): video-polygraphic features and long-term outcome. AB - OBJECTIVE: The aim of this study was to investigate the video-polygraphic features and the long-term outcome of epilepsy in two patients with startle epilepsy associated with infantile hemiplegia (SEIH). MATERIAL AND METHODS: Two patients (patient 1: a 14-year-old girl; patient 2: a 17 year-and-half-year-old girl), with hemiparesis and moderate mental retardation, underwent a full clinical and neurophysiological examination with video-polygraphic monitoring and recording of startle-evoked seizures. The follow-up was 9 years from epilepsy onset in patient 1, and 8 years from epilepsy onset in patient 2. RESULTS: Firstly, video-polygraphic recordings of startle-evoked seizures, triggered by unexpected auditory stimuli, showed tonic asymmetrical postures with ictal EEG characterized by an abrupt and diffuse electrodecremental pattern or a seizure discharge predominant over the vertex and anterior regions controlateral to the posturing limbs. Electromyogram recording showed a prevalent involvement of proximal muscles with a concomitant tachycardia and apnoea. In particular, in patient 1 ictal heart rate was high, with persisting tachycardia for 60-120 s after the end of seizures. Secondly, a high seizure frequency persisted throughout the course of the disease, as seizures were medically refractory to all currently available anti-epileptic drugs. CONCLUSIONS: The long-term outcome of epilepsy in SEIH, with constantly high seizure frequency, suggests an early surgical intervention, avoiding years with unsuccessful drug treatments and poor quality of life. PMID- 19809942 TI - International Journal of Sports Medicine - ethical standards in sport and exercise science research. PMID- 19809944 TI - Embryology and epidemiology of microtia. AB - The auricle derives from six hillocks arising from the first and second branchial arches. Different hillocks give rise to different parts of the pinna. In the course of embryonic development, the auricle migrates postero-cranially as the mandible enlarges. Auricular malformations, such as microtia, are thought to be related to cell death of the first and second arch derivatives. The prevalence and characteristics of microtia vary in different populations. The prevalence ranges from 0.83 to 17.4 per 10,000. Microtia is more common in males, and right sided dominance varies from 57 to 67%. The prevalence of aural atresia or stenosis varies from 55 to 93%. Microtia has been associated with numerous risk factors including race and gender. Genetic factors are likely to have an effect at least in some patients with microtia. PMID- 19809945 TI - Basic techniques in autogenous microtia repair. AB - Although several techniques can be used for microtia repair, some standard procedures have been developed over the past few decades. In specialized centers, these techniques are performed most frequently. They include two to four operative steps using autogenous rib cartilage for the framework and local skin for its tegument. We have been using this technique presented here in more than 800 cases over the past two decades with good and mostly stable results. PMID- 19809946 TI - Learning to perform ear reconstruction. AB - Learning how to perform ear reconstruction is very difficult. There are no standardized teaching methods. This has resulted in many ear reconstructions being suboptimal. Learning requires a major commitment by the surgeon. Factors to be seriously considered by those considering performing this surgery are (1) commitment, (2) aptitude, (3) training methods available, (4) surgical skills and experience, and (5) additional equipment needs. Unless all these factors are addressed in a surgeon's decision to perform this form of reconstruction, the end result will be compromised, and patient care will not be optimized. It is hoped that considering these factors and following this approach will result in a higher quality of aesthetic result. The future of ear reconstruction lies in the use of advanced digital technologies and tissue engineering. PMID- 19809947 TI - The learning curve in microtia surgery. AB - Reconstruction of the auricle is known to be complex. Our objective was to evaluate the improvement of the outcome of the lobulus-type microtia reconstruction. Patient satisfaction was also evaluated. There are no previous reports of the learning process in this field. Postoperative photographs of 51 microtia reconstructions were assessed and rated by a panel made up of six surgeons. The ratings were gathered to generate learning curves. Twenty-two patients assessed the outlook of their reconstructed ears, and the results were analyzed as a self-assessment group. The reliability of the rating by a panel was tested by intraclass correlations. There is a highly significant increasing trend in learning ( P = 0.000001). This trend is not constantly upward, and the steady state was not reached during the study. In the self-assessment group, females were significantly more critical than males ( P = 0.014). Intraclass correlation for six panel members was 0.90, and the rating was considered reliable. Thus, a long and gentle learning curve does exist in microtia reconstruction. To secure good quality and continuity, centralization of the operations and trainee arrangements are highly advisable. Outcomes of plastic surgery can reliably be rated by an evaluation panel. PMID- 19809948 TI - Reducing the morbidity involved in harvesting autogenous rib cartilage. AB - Although the use of autogenous cartilage is the gold standard in auricular reconstruction, its main disadvantage is the morbidity due to harvesting the cartilage. This includes postoperative pain, visible scar, and possibly asymmetry and reduced stability of the thorax. To reduce all of these drawbacks, we describe some modifications that reduce pain to a low tolerable level, hide the scar invisibly in the submammary fold in females, and induce regeneration as well reestablish stability of the rib defect. PMID- 19809949 TI - Treatment of unwanted hair in auricular reconstruction. AB - In many microtia patients, the hairline is lower than ideal. Despite this, it is essential to position the reconstructed ear in the correct place. Here is a series of tips and tricks to deal with unwanted hair on the skin that covers an autogenous tissue reconstruction. Replacement of the skin with a fascial flap and skin graft remains the mainstay of treatment for a very-low-hairline case. Surgical removal of hair on the helical rim during the release procedure is also described. Laser depilation, surgical electrolysis, and even shaving techniques are also discussed. PMID- 19809950 TI - Ear reconstruction using a porous polyethylene framework and temporoparietal fascia flap. AB - Ear reconstruction is a difficult procedure requiring a framework and soft tissue covering. The traditional method uses a rib cartilage framework placed beneath scalp skin. This method has been used for 50 years despite inherent problems with both harvesting rib cartilage and using scalp for coverage. The authors describe a method using a porous polyethylene (PPE) framework covered by a large temporoparietal fascia (TFP) flap raised with the underlying subgaleal fascia (SGF). The entire implant is covered by the two-layered flap, which can be raised without any scalp incision. The skin grafts applied to the covered implant lie on the SGF. The trilaminar structure of the SGF allows the skin to move independently over the implant, resisting shear forces and reducing the probability of implant exposure. Ear reconstruction using the PPE framework was performed on 786 ears over an 18-year period. Initial complications were common. With improved implant design and complete coverage of the implant with both the TPF and SGF, exposure rate dropped to 7% with a 12-year follow-up. Implant fractures decreased to less than 3%. The PPE/TPF method allows earlier ear reconstruction in children with minimal scarring and discomfort. The reconstructed ear can closely mimic the shape and projection of the natural contralateral ear in fewer stages and with a shorter learning curve. PMID- 19809951 TI - Ear epistheses as an alternative to autogenous reconstruction. AB - An ear episthesis is an artificial substitute for the auricle. The term EAR PROSTHESIS is used synonymously. The breakthrough came with the introduction of the modern silicones and their colorings. Although there are still indications for noninvasive methods of retention such as medical adhesives, the best and most reliable method of fixation is by bone anchorage. Long-lasting osseointegration with reaction-free skin penetration can be achieved with titanium implants. The first system used extraorally was the Branemark flange fixture. Later, different solitary titanium implants were introduced, such as the ITI system. A different strategy used the titanium grids (Epitec) or plates (Epiplating) derived from osteosynthesis systems. These systems are fixed subperiosteally with several bone screws and are therefore also labeled as grouped implants. With these modern developments, secure retention can be achieved also in unfavorable anatomic situations. The grouped systems are resistant to torque with abutment insertion. The latest development is the subcutaneously implanted double magnet without skin penetration. The advantages of implant retained ear epistheses include optimal camouflage, predictable cosmetic results, fast rehabilitation, no donor site morbidity, and early detection of tumor recurrence. Depending on the clinical setting, prosthetic rehabilitation may be more than just an alternative to plastic reconstructive surgery. PMID- 19809952 TI - Special reconstruction techniques for special circumstances. AB - Auricular reconstruction is a challenging, mostly multistaged procedure that requires many different techniques. Whereas standard techniques have been developed for the standard severe, third-degree dysplasia, we are often confronted with severely scarred patients due to excess trauma or prior operations. These special situations need a variety of special techniques; some of those are described here to be considered as alternative options. PMID- 19809953 TI - [Gender -- a theme in gastroenterology?]. PMID- 19809954 TI - [Are there gender-related differences in the therapeutic management of patients suffering from inflammatory bowel disease? Subgroup analysis of a prospective multicentre online-based trial]. AB - INTRODUCTION: The most frequently prescribed medications for patients suffering from inflammatory bowel disease (IBD) in the Rhein-Main region of Germany are aminosalicylates and corticosteroids irrespective of the disease activity. In contrast, immunomodulators only play a marginal role. As anti-TNF therapy is very costly, it is prescribed in outpatient services of hospitals rather than in gastroenterological practices. AIM: The aim of this study was to evaluate possible gender-related differences in the therapeutic management of IBD patients treated in the Rhein-Main region of Germany. METHODS: Data records about past medical history, disease status, laboratory values and medical treatment of outpatients of 10 gastroenterological practices and 3 hospitals were collected from November 1st 2005 to July 31st 2007 and analysed with regard to gender related differences in therapy and disease management. RESULTS: Overall, no statistically significant difference in gender-specific medical treatment could be observed in the study cohort. However, detailed analyses revealed, that 1. women suffering from IBD, who are treated in outpatient services of hospitals, are more often under immunosuppressants, irrespective of disease activity, 2. in gastroenterological practices less than 3 % of patients are prescribed any immunosuppressive therapy (vs. 17 % [men] und 42 % [women] in hospital outpatient services), and 3. anti-TNF therapy is applied more frequently in men as compared to women in hospital outpatient services in both remission and active disease. CONCLUSION: This study discloses the gender-specific differences in the therapeutic management of IBD patients in a congested urban area in Germany. Further studies are required to confirm the tendencies detected. PMID- 19809955 TI - [Colon cancer risk in persons at familial or hereditary risk aged < 55 years]. AB - BACKGROUND: The question whether persons at familial or hereditary risk differ in terms of absolute, relative, or cumulative risk for colorectal cancer or not is of importance for the estimation of the potential of early detection of colorectal cancer in persons with familial or hereditary risks. METHODS: Based on the results of a systematic literature search on absolute, relative, and cumulative risks of familial and hereditary disposition for colorectal cancer as well as actual German tumour incidence data, projections were conducted. RESULTS: The absolute risk for colorectal cancer in familial risk persons identified by means of a questionnaire is increased by a factor of 2 - 4 depending on the age at questioning, the age of the family member at cancer diagnosis and number of family members with colorectal cancer. Their absolute colorectal cancer risk equals that of persons without this risk who are 10 to 15 years older. Persons with hereditary risk show an increase in risk by a factor of 8 - 80. CONCLUSION: Persons aged 40 to 45 years with a familial risk constellation show a risk for colorectal cancer that equals the risk of 55- to 59-year-old persons from the general population. Therefore, the legal right for screening colonoscopy should be extended to the persons at risk aged 40 to 45 years. Persons suspected for hereditary risk should have a genetic counselling and, in case of germ mutation, a colonoscopic surveillance according to the actual guidelines. PMID- 19809956 TI - [The first case of anisakiasis acquired in Austria]. AB - Anisakiasis is caused by a fish parasite of the Nematode family. This kind of rare helminthozoonosis can mainly be found in countries where consumption of raw fish is traditionally high like Japan, the Netherlands, Pacific Islands, South Europe, Scandinavia, USA, and Canada. Man is the wrong hoste. Clinical manifestation depends on the localisation of penetration in the GI tract. In Japan, predominantly the stomach is affected in 97 % of cases, probably due to hypo- and achlorhydria; whereas mainly intestinal anisakiasis occurs in Europa. We report on a 67-year-old male patient with a gastric infestation of anisakiasis. The patient was on proton pump inhibitor which migh have caused the localisation of the infestation. The anisakis was an accidental endoscopic finding in a patient for control of an H. p.-positive gastric ulcer. Otherwise the patient was free of pain. The helminth (larva III) was endoscopically extracted. Thereafter, the patient remained in good health. Anisakis serology as well as repeated differential blood counts were without finding. The uneventful medical history and the normal blood findings indicate that our patient had a very early stage of infestation of anisakiasis. The patient reported no stay outside of Austria within the last years. However, he consumed on a regular basis "rolled pickled herring" produced by a well-known Viennese company for canned fish. This is the first documented case of this rare helminthozoonosis acquired in Austria. PMID- 19809957 TI - Primary actinomycosis of the liver mimicking malignancy. AB - A 71-year old women presented with fever, a significant loss of body weight and abdominal pain in the upper right quadrant since approximately six months. Abdominal ultrasonography and magnetic resonance imaging (MRI) showed an irregularly shaped, inhomogeneous and hypointense lesion of the right liver lobe (6 x 8 cm in segment 7 and 8) with multiple satellite lesions. Irregular shape, hypovascular presentation during gadolinium enhancement, hypointensity in T 1 weighted images and dilation of peripheral bile ducts were suggestive for cholangiocarcinoma or metastasis. However, histological investigations revealed a rare case of primary actinomycosis of the liver which was successfully treated with antibiotics. PMID- 19809958 TI - [Reflux disease and Barrett's oesophagus - are there gender-specific differences?]. AB - Studies of the prevalence of gastroesophageal reflux disease (GERD) have reported that male gender is an independent risk factor especially for erosive reflux disease (ERD). Non erosive GERD (NERD) is more common in females. The rate of prevalence and severity of reflux symptoms increase in females with age, while among men it peaked between 50 and 70 years and thereafter declined. The gender effect may be caused by differences in parietal cell mass between males and females. Barrett's esophagus is a major complication of gastroesophageal reflux disease and is associated with 30-125 times increased risk of developing carcinoma. Most studies have found gender differences with females significantly less likely to have Barrett's esophagus with a 20-yr age shift between the parallel age specific prevalence curves, for males between the ages of 20 and 59 yr and for females between the ages of 20 and 79 yr. Male predominance for ERD as a precursor to Barrett's esophagus may be partly explain the greater male/female sex ratio for Barrett's esophagus. Female sex hormones may play a protective role. Knowledge of gender-specific differences of reflux disease and Barrett's esophagus may be helpful to improve surveillance and screening strategies, although distinct recommendations are lacking so far . PMID- 19809959 TI - [Inflammatory bowel disease and pregnancy]. AB - Inflammatory bowel disease (IBD) affects mainly young people who are often in the process of family planning. Insecurity of patients and their primary physicians concerning the disease course and the appropriate medication is often followed by a decision against the pregnancy. With this overview these central issues will be discussed and practical guidelines will be provided, thus encouraging both sides, that IBD and pregnancy is compatible. PMID- 19809960 TI - [Value of colour Doppler ultrasonography in relation to clinical pretest probability in giant cell (temporal) arteritis]. AB - BACKGROUND: Colour Doppler sonography (CDS) is an established technique in the diagnosis of giant-cell (temporal) arteritis (GCA). The predictive value of its diagnostic criteria for GCA (halo sign or stenosis) is related to the pretest probability (PTP), a measure of probability of presence of a target disease before the result of a diagnostic test is known. PATIENTS AND METHODS: A total of 182 (average age 69 years, 69% women) patients of the Rheumatology Center Baden Wurttemberg were investigated. Based on the diagnostic criteria of the American College of Rheumatology (ACR) they were assigned to one of three groups, before a CDS was performed: group 1 (n= 139) patients with "isolated" polymyalgia rheumatica (PMR) and a low PTP for GCA; group 2 (n=19) patients with intermediate PTP and nonspecific headache and fewer than three ACR criteria for GCA); and group 3 (n=224) patients with a high PTP and new headache loclized to the temporal artery and at least three ACR criteria for GCA. RESULTS: The halo sign (periluminal dark halo) of more than 0.3 mm was present in 26% of group 1. 42% of those in group 2 and 83% of those in group 3. A stenosis or occlusion of the temporal artery was present in 3.5% (group 1), 5% (group 2) and 46% (group 3), respectively. 3 of 24 patients of group 3 also had a stenosis of the axillary or brachial artery. Concordance between clinical criteria and CDS (normal CDS in patients with PMR but no headache or abnormal CDS and clinically suspected BCA was found in 123 of 182 patients (67.5%). In these patients biopsy of the temporal artery ("gold standard" for the diagnosis of GCA) was not recommended. Temporal artery biopsy was, however, recommended in all patients with discordant findings (abnormal CDS with PMR but no headache or normal CDS with clinically suspected GCA, and also those with intermediary PTP (32%). A biopsy was performed in 42 of these patients after informed consent had been obtained. This demonstrated vasculitis in 11 of 25 patients with PMR (PPV in group 1: 0.44). But biopsies were negative in all four patients with clinically suspected GCA and normal CDS (NPV in group 3:1). In the intermediary group biopsy demonstrated vasculitits in 5 of 6 patients with an abnormal CDS (PPV 0.63), while 4 of 5 patients with a normal CDS had a normal biopsy (NPV 0.8). CONCLUSION: Taking into account pretest probability, an RCA can be accurately diagnosed or excluded by CDS in two thirds of patients without biopsy. When performed by an experienced investigator CDS is a basic part in the diagnosis of CDA. PMID- 19809961 TI - [Anticoagulation during pregnancy following an artificial heart valve replacement]. AB - HISTORY AND CLINICAL FINDINGS: A 23-year-old woman had received a mechanical bileaflet mitral valve prosthesis because of severe mitral valve insufficiency caused by an acute bacterial endocarditis with vegetations. One year after the operation the patient suffered on two miscarriages under oral anticoagulation by phenprocoumon. Present, she was referred to our center with the question of conversion to low molecular weight heparine because of continued yearning for a baby. INVESTIGATIONS: At admission the woman was in good general and nutritional condition. Echocardiography showed a regular prosthetic function. Blood analysis, electrolyte parameters and enzyme values were normal, further laboratory investigations revealed a factor-V-Leiden-mutation. A chromosomal analysis detected no aberrations. TREATMENT AND COURSE: The oral anticoagulation by phenprocoumon was switched to subcutaneous low molecular weight heparine in therapeutical dosage. Anti-factor-Xa-activity was controlled at regular intervals. Further pregnancy was uneventful for both, mother and child. A healthy infant was born by caesarean section at 40 (th) week of gestation. CONCLUSIONS: Treatment with anticoagulation by phenprocoumon is indispensable for mechanical heart valve protheses. Conversion to low molecular weight heparine is possible in patients who insistent request to conceive. The anticoagulation by low molecular weight heparine avoids teratogenic effects during pregnancy because the placenta is impermeable to that heparin. Furthermore, prophylaxis of thromoses by low molecular weight heparine is probably in almost the same manner as by phenprocoumon. PMID- 19809962 TI - [Diagnosis of fever of unknown origin (FUO). Positron emission tomography/computed tomography (PET-CT)]. AB - Nuclear medicine imaging is now well accepted for the localization of septic foci. But in patients the results of infection scintigraphy, radiology and ultrasound remain unsatisfactory in the diagnosis of fever of unknown origin (FUO). In contrast to septic infections, patients with FUO - mostly in elderly patients - tend to have such conditions as occult tumours, atypical pneumonia, hematoblastosis, malignant lymphomas. (18)F(Fluor-18)-Fluordeoxyglucose-PET ((18)F-FDG PET) has made it possible to localize symptomatically occult changes with a high diagnostic accuracy and to achieve differentiation between benign and malignant changes. PMID- 19809963 TI - [67-year-old man with symptoms of influenza and a swollen neck]. PMID- 19809964 TI - [Fingolimod - a new immunomodulator]. AB - Fingolimod is a derivative of the naturally occurring immunosuppressive substance myriocin, with structural similarity to sphingosine, a ubiquitous sphingolipid. It acts as an immunomodulator interfering with the egress of T and B lymphocytes from secondary lymph organs, which results in lymphopenia. A phase II study in patients with multiple scerosis showed a significant reduction in annual relapse rate and lesions in magnetic resonance imaging with an acceptable rate of side effects. An advantage of fingolimod compared to interferon beta, glatirameracetate and natalizumab is its oral availability. Definitive assessment of the compound has to await the results of trials currently being performed. PMID- 19809965 TI - ["TRECID", TNFalpha related chronic inflammatory diseases - a new multiple diseases bridging concept]. AB - The pro-inflammatory cytokine TNF alpha (TNF) has a key position in the pathogenesis of various infectious and inflammatory diseases. Clarification of its pivotal role in the pathogenesis of rheumatoid arthritis, spondyloarthritis, uveitis, psoriasis and inflammatory bowel disease has resulted in the successful development of TNF- blocking therapies, which have disease-modifying properties that exceed the effects of conventional therapeutic options. For this reason data on the concurrence of several chronic inflammatory diseases have led to the hypothesis of common pathogenetic processes of cytokine dysregulation. The acronym TRECID describes this concept of "TNF RElated Chronic Inflammatory Diseases". Physicians of different specialties have integrated new therapeutic options with TNF-blocking therapies into their strategies for the management of the affected patients. Thus the concept of TRECID can be regarded as a role model for a dynamic, interdisciplinary cooperation based on shared pathophysiological aspects. PMID- 19809966 TI - A short cervical length in pregnancy: management options. AB - Measuring cervical length using transvaginal ultrasonography is a useful tool to predict the risk of preterm birth in low- and high-risk pregnancies. Management of a short cervix poses a significant dilemma for clinicians. Different management plans have been proposed and studied, with mixed results in different clinical settings. This article reviews the various management options in the different patient subpopulations and proposes a scheme for management once a short cervix is identified. PMID- 19809967 TI - [Sonography of the liver following CT staging of patients with lung cancer: a retrospective analysis]. AB - PURPOSE: Liver lesions are frequently detected in the CT staging of lung cancer patients and may require further investigation. The aim of our study was to assess the value of an ultrasound (US) examination of the liver in addition to routine CT staging. MATERIALS AND METHODS: In this retrospective study we included 174 consecutive patients with lung cancer who underwent US of the liver in addition to contrast-enhanced CT of the thorax and upper abdomen. The reports of the examinations were evaluated for the presence of liver lesions. Based on CT and US standard criteria, liver lesions were grouped into unequivocal cysts, hemangiomas, metastases and undefined lesions. RESULTS: With CT, liver lesions were detected in 56 / 174 patients (32 %). These included 24 cysts in 11 patients, 2 hemangiomas in 2 patients and 18 patients with liver metastases. In 31 patients, 66 small (< 1.5 cm) hypodense lesions were detected, which could not be further defined by CT. Using US, 21 of these 66 liver lesions were confirmed as benign (cysts, hemangiomas), and two lesions were diagnosed as metastases. In 2 patients US revealed metastases that were not visible on the CT scans. CONCLUSION: The study demonstrates that a complementary US of the liver in patients with lung cancer may reveal information relevant for treatment. Therefore, liver US may play an important role in the staging of lung cancer. When equivocal small liver lesions are detected with CT, a complementary US examination may help to diagnose these lesions or detect metastases not visible on the single-phase staging CT of the liver. PMID- 19809968 TI - Superior cerebellar artery - a potential pitfall in transcranial posterior cerebral artery insonation. AB - PURPOSE: Transcranial color-coded duplexsonography (TCCD) of the posterior cerebral artery (PCA) is commonly performed in an axial midbrain insonation plane. The proximal PCA is easily identified as a vessel, originating from the top of the basilar artery (BA). However, the initial segment of the superior cerebellar artery (SCA) also originates from the top segment of the BA and runs in parallel. Therefore, it is possible to confuse the two vessels. The aim of our study is to anatomically differentiate the proximal PCA and SCA by means of TCCD in the coronal plane using a transtemporal approach and to analyze and compare their flow responses during a visual stimulus paradigm. MATERIALS AND METHODS: 60 subjects without a relevant vascular pathology and with a good temporal bone window were examined. The distal BA, the proximal PCA and the SCA were insonated using a coronal insonation plane and identified by their anatomical relationship. The visible length was measured and the blood flow velocities were recorded at rest and during visual stimulation. RESULTS: The identification and differentiation of both vessels was successful in 101 of 120 hemispheres (84.2 %). The PCA systolic flow velocities at rest and during visual stimulation were 58.4 +/- 11 and 71.8 +/- 13 cm/sec (23.6 % increase), while the SCA velocities were 50.7 +/- 12 and 53.5 +/- 13 cm/sec, respectively (5.8 % increase). The difference was significant (p < 0.001). CONCLUSION: Using the coronal transtemporal insonation approach, TCCD makes it possible to identify and distinguish the proximal PCA and SCA. Our study is the first to provide a systematic ultrasound assessment of the SCA and includes data on normal flow velocities and reactivity to a visual stimulus. PMID- 19809969 TI - Taxomyces andreanae: a presumed paclitaxel producer demystified? AB - The 1990s brought an abundance of reports on paclitaxel-producing endophytes, initially heralded as a discovery having tremendous implications for cancer therapy. As the vision of large-scale fermentation tanks producing vast quantities of relatively inexpensive paclitaxel and novel taxanes has faded and has been replaced by controversial silence, we carried out an in-depth investigation of Taxomyces andreanae - the very first presumed endophytic synthesizer of the diterpenoid. On one hand, metabolic profiling by means of chromatographic, spectroscopic and immunoenzymatic techniques predominant in literature was taken up. On the other, the experimental procedure was brought to an alternative, previously unattempted level aiming at revealing the genetic background of paclitaxel biosynthesis in the endophyte. The profound PCR-based screening for taxadiene synthase (TXS) - a gene unique to the formation of the primary taxane-skeleton, as well as phenylpropanoyl transferase (BAPT) encoding for the catalyst of the final acylation of the core structure rendering the ultimate efficacy of the drug, confirmed the molecular blueprint for paclitaxel biosynthesis to be an inherent genetic trait of the endophyte. However, as the thorough metabolic analysis of Taxomyces andreanae commercial isolate brought no confirmation of endophytic paclitaxel production even after considerable up scaling endeavors, we postulate that proclaiming the strain "a fungus factory for Taxol" might have been premature. PMID- 19809970 TI - Transformed cell suspension culture of Galphimia glauca producing sedative nor friedelanes. AB - The Mexican species GALPHIMIA GLAUCA (Cav.) Kuntze (Malphigiaceae) synthesises a family of sedative and anxiolytic nor-secofriedelanes, designated as galphimines. These active principles accumulate at low concentration in the aerial parts of plants from wild populations. Transformed calluses and cell suspension cultures of this species were established in order to induce a greater production of nor friedelanes. The cell suspension line GgBa was selected and grown over a period of two years of continuous subculturing in MS nutrient medium in the absence of growth regulators. PCR and Southern blot analyses were employed in order to confirm that the ROL A gene had been integrated into the plant genome. Batch cultures of the GgBa cell line were grown over a 32-day period and first-order growth kinetics was observed, reaching a specific growth rate (micro) of 0.13 d ( 1). The production of glaucacetalin A ( 10), a triterpenoid related to the known galphimines, was quantified in the nutrient medium by HPLC. The transformed cell suspension culture GgBa also synthesised a novel nor-friedelane, given the name glaucacetalin D ( 13). High-resolution spectroscopic and spectrometric techniques were employed to elucidate the structure of 13. This triterpene has never been observed in wild plant tissues or in other IN VITRO cultures. Maslinic acid ( 14) was identified in cell biomasses. The triterpene production of the cell line GgBa was as follows: glaucacetalin A, 2.7 mg/L; glaucacetalin D, 2.9 mg/L and maslinic acid, 2.4 mg/g dry weight. The sedative activity of compounds 10 and 13 was demonstrated in ICR mice by using the sodium pentobarbital-induced hypnosis model. No cytotoxicity of 10 and 13 was exhibited against KB, MCF-7 and HF6 human cancer cell lines. PMID- 19809971 TI - Root colonization by symbiotic arbuscular mycorrhizal fungi increases sesquiterpenic acid concentrations in Valeriana officinalis L. AB - In some medicinal plants a specific plant-fungus association, known as arbuscular mycorrhizal (AM) symbiosis, increases the levels of secondary plant metabolites and/or plant growth. In this study, the effects of three different AM treatments on biomass and sesquiterpenic acid concentrations in two IN VITRO propagated genotypes of valerian ( VALERIANA OFFICINALIS L., Valerianaceae) were investigated. Valerenic, acetoxyvalerenic and hydroxyvalerenic acid levels were analyzed in the rhizome and in two root fractions. Two of the AM treatments significantly increased the levels of sesquiterpenic acids in the underground parts of valerian. These treatments, however, influenced the biomass of rhizomes and roots negatively. Therefore this observed increase was not accompanied by an increase in yield of sesquiterpenic acids per plant. Furthermore, one of the two genotypes had remarkably high hydroxyvalerenic acid contents and can be regarded as a hydroxyvalerenic acid chemotype. PMID- 19809972 TI - Benzopyranones and their sulfate esters from Pelargonium sidoides. AB - Investigation of the benzopyranones from the roots of PELARGONIUM SIDOIDES led to the identification of 15 benzopyranones, among them the novel compounds 12 and 14. Furthermore, benzopyranones 4 and 15 were detected in PELARGONIUM SIDOIDES for the first time. Structure determinations were performed by one- and two dimensional NMR spectroscopy. An HPLC system for the discrimination of these benzopyranones has been developed. PMID- 19809973 TI - Impact of thymol in thyme extracts on their antispasmodic action and ciliary clearance. AB - Thyme is a herb with broncholytic und secretomotoric effects. Its activity on beta (2) receptors as a possible mechanism of action was demonstrated. Major components are thymol and carvacrol which are claimed to be responsible for its effects and, therefore, used for standardization in the German pharmacopoeia (0.03 % phenols calculated as thymol). Our aim was to investigate the impact of thymol by using thyme extracts with either normal or extremely low thymol concentrations (< 0.005 % or > 0.038 %). The antispasmodic effect on smooth muscles of the trachea and the ileum and the effect on ciliary activity (respiratory clearance) were investigated. In addition, pure thymol and carvacrol were investigated separately and in spiking experiments. Thymol and carvacrol had a concentration-dependent antispasmodic effect in the rat trachea either being stimulated by acetylcholine, K (+) or Ba (++). The same result was observed with respect to the increase of mucociliary transport in mice. Extracts with very low thymol contents are effective in all models used except acetylcholine-induced rat ileum contraction. When thyme extracts with normal thymol contents or with very low thymol contents were compared, the extract with normal thymol contents was more effective, both as a relaxant (rat ileum) and as an antispasmodic compound (rat trachea contraction induced by either acetylcholine, Ba (++) or K (+)) and in ciliary transport experiments. Thyme extracts with very low thymol contents (practically free of volatile oil) were equally effective with respect to endothelin effects. When an extract with very low thymol contents is spiked with increasing concentrations of thymol, a concentration-dependent increase concerning the antispasmodic effect (Ba (++)-induced trachea contraction) is observed. In conclusion, the data show that in various models of antispasmodic effect (ileum and trachea) and by measuring ciliary activity, thymol (and carvacrol) is (are) active, although other not identified components of thyme extract appear to be very important as well, since extracts with very low thymol contents are active. On the basis of these results the standardization on thymol alone appears not to be justified. PMID- 19809974 TI - [Needlestick injury--risk and prevention of blood borne infections]. PMID- 19809975 TI - [What is your diagnosis? Calcified subperiosteal hematoma]. PMID- 19809976 TI - [Secondary forms of diabetes mellitus]. AB - Diagnosis and treatment of secondary forms of diabetes are different from the common management of type 1 or 2 diabetes. Furthermore, they are often associated with concomitant disease that are important for prognosis. Case reports are presented in order to describe secondary diabetes associated with hemochromatosis, cystic fibrosis, pregnancy, lipodystrophy, endocrinopthies, MODY and mitochondrial diabetes. PMID- 19809977 TI - [Osteoporosis: diagnostic assessment]. AB - Osteoporosis is a generalized disease of bone that increases the fracture risk. Among the multiple factors involved in osteoporosis, some, but not all, are related to bone mass. Although bone mineral density (BMD) measurement is specific for detecting high-risk individuals, it misses a notable proportion of individuals who have clinical or epidemiological risk factors for osteoporotic fractures. Therefore, composite scores that rely both on BMD and on validated clinical risk factors have been developed. The Fracture Risk Assessment Tool (FRAX) was designed to predict the 10-year probabilities of sustaining a major osteoporotic fractures or a hip fracture. The cutoff value beyond which treatment should be initiated needs to be determined, based not only on clinical criteria, but also on economic considerations. PMID- 19809978 TI - [Prostate cancer: a review of therapeutic strategies]. AB - Every third new cancer in men is a prostate cancer and we expect a steadily increase of the incidence in the next years. The median age at diagnosis is 68 years and we see about 4000 new prostate cancer patients every year. Approximately 1 in 7 men will eventually be diagnosed with PC. Stage dependent treatment strategies may include surgery, radiotherapy and systemic treatments like hormonal ablation and chemotherapy. The aim of this article is to give an overview of modern treatment strategies. PMID- 19809979 TI - [Obesity surgery--useful knowledge in indication and follow up]. AB - In the last years, obesity has become one of the main problems of health care systems in Western countries. Among morbid obese patients, four out of five will develop comorbidities doubling the mortality risk in women and increase the numbers in men at a threefold risk. According to evidence based guidelines, nowadays surgery is the best and most effective treatment resulting in excellent long-term weight loss, reduction of comorbidities while extending expectation of life. A sound indication is the most important step for successful surgery. In this paper we focus on Swiss regulations and some special indications which have to be taken into consideration. After bariatric surgery clinical follow up on a regular basis is also of great importance. Furthermore, we explain typical mechanical and nutritional complications after different types of surgery and give some recommendations. PMID- 19809980 TI - [New drugs for control of blood glucose in type 2 diabetes: summary of the NICE guidelines]. PMID- 19809981 TI - [Abdominal mass and colicky pain in a 16-years-old patient]. AB - We report about a 16-years-old patient with a newly developed palpable mass and pain in the epigastrium. On computed tomography, a trichobezoar was diagnosed. Complementary history, gathered retrospectively, revealed trichophagia and swallowing of chewing gum. Gastroscopic disintegration of the trichobezoar was unsuccessful and it therefore had to be removed by open surgery. PMID- 19809982 TI - [Intensive blood glucose control in patients with type 2 diabetes lowers the risk of cardiovascular events (very) moderately]. PMID- 19809983 TI - [Intensive blood glucose control in type 2 diabetic patients does not have a significant effect on macrovascular complications]. PMID- 19809984 TI - [A single infusion of zoledronic acid (Zometa) for prevention and treatment of steroid-induced osteoporosis]. PMID- 19809988 TI - Chronic kidney disease certification process manual by the Italian Society of Nephrology (SIN): part II: programme management and clinical information management. AB - This is the second part of a document describing a voluntary certification process based on Joint Commission International (JCI) criteria developed by the Italian Society of Nephrology (SIN) and JCI representatives. In the first part we discussed standards for clinical care delivery and performance measurements related to chronic kidney disease care. Herein (Part II), we complete the description of Performace measurements and CKD care by describing issues related the management and clinical information management. PMID- 19809989 TI - Chronic kidney disease among Indigenous populations: considerations for effective and ethical research. AB - Chronic kidney disease (CKD) is a well-documented and growing problem among Indigenous populations in North America and Australia. Further, urgent research is needed to develop appropriate interventions to slow development and progression of CKD and to improve outcomes in Indigenous* communities affected by the burden of kidney disease. For effective research to occur, researchers need to develop and maintain a multifaceted and collaborative approach to working with Indigenous research subjects and their communities. We review two fundamental concepts or paradigms which may cause misinformation or confusion in conducting health research in Indigenous populations. First, we examine systems of health knowledge and discuss the divergences between investigator and Indigenous perspectives, and how they interface in a research context. Secondly, we review the concept of research methods for Indigenous populations, to highlight ways to develop a collaborative and culturally inclusive health research process. PMID- 19809990 TI - Hypertension in the elderly: new and old evidence that treatment may benefit the old. AB - Hypertension is prevalent and remains an important risk factor in elderly and very elderly. Randomized controlled outcome trials have shown benefit of antihypertensive treatment in patients with systolic diastolic hypertension and in patients with isolated systolic hypertension, aged 60 years and over. More recently benefit has also been shown in octogenerians with hypertension. Overall there is no strong evidence that protection against major cardiovascular events afforded by different drug classes varies substantially with age. Finally, blood pressure lowering therapy also improves prognosis in hypertensive patients with type 2 diabetes. PMID- 19809991 TI - Managing dry weight and hypertension in dialysis patients: still a challenge for the nephrologist in 2009? AB - Hypertension prevalence is high in both hemodialysis and peritoneal dialysis patients. Among several possible mechanisms leading to hypertension in chronic kidney disease, the most important is the positive balance of sodium and accumulation of extracellular fluid causing cardiovascular remodeling. Hypertension may provoke de novo left ventricular hypertrophy and cardiac failure in dialysis patients and is also associated with a higher risk of stroke. The relationship between hypertension and mortality is unexpected, with better survival in hypertensive patients in short-term studies in which heart failure associated with low blood pressure and high death rate confounds the effect of hypertension on death risk. Higher mortality is found in hypertensive patients surviving beyond 2 years under dialysis treatment. The non pharmacological treatment of hypertension - the dry weight method - is efficient to correct hypertension when applied. A recent randomized controlled trial has confirmed its efficiency in lowering blood pressure. A low-salt diet is part of this strategy limiting the interdialytic weight gain and enhancing tolerance to ultrafiltration. Moreover, according to recent meta-analyses, the prescription of antihypertensive medications is associated with better survival. Their appropriate use remains to be studied. In conclusion, the important challenge for the nephrologist in 2009 is to efficiently correct extracellular fluid overload and its consequences, with the goal of improving the high burden of cardiovascular mortality among dialysis patients. PMID- 19809992 TI - Statin treatment for dyslipidemia in chronic kidney disease and renal transplantation: a review of the evidence. AB - Patients with chronic kidney disease (CKD) have significantly increased risks of cardiovascular (CV) morbidity and mortality. Dyslipidemia is a common disorder in CKD patients. CKD patients have a different lipid profile with increased atherogenic lipid fractions, and serum low-density lipoprotein cholesterol (LDL C) levels may underestimate the atherogenic effect of LDL-C in these patients. Dyslipidemia may contribute to the increased CV morbidity and mortality, and to the progression of kidney disease in CKD patients. Currently, statins are the pharmacologic intervention of first choice, if lifestyle changes fail adequately to lower LDL-C levels in the setting of normal or moderately elevated triglycerides. Statins have been extensively studied in a large variety of patient populations and have proven efficacy in the treatment of dyslipidemia, and in reducing CV mortality. Although much evidence supports the CV benefits of statins in patients with normal renal function, there are contradictory results for the beneficial effect of statin therapy on CV morbidity and mortality in CKD patients. While post hoc subgroup analyses of multiple randomized trials support statin use in early CKD patients, the only randomized trial conducted in diabetic dialysis patients found no evidence of benefit in overall mortality. Post transplant there is some definite CV benefit, albeit in a patient cohort selected to be at reduced CV risk by virtue of being eligible for organ transplant. The results from the AURORA and SHARP studies are awaited anxiously. PMID- 19809993 TI - Beneficial effect of chlorambucil in steroid-dependent and cyclophosphamide resistant minimal change nephrotic syndrome. AB - BACKGROUND: Little information is available on the effect of second cytotoxic therapy in steroid-dependent children with minimal change nephrotic syndrome (MCNS). METHODS: Response to second cytotoxic therapy and side effects were reviewed in 33 steroid-dependent and cyclophosphamide-resistant children with MCNS who received chlorambucil (n=11, group 1) or cyclophosphamide (n=22, group 2). RESULTS: Age at onset of nephrosis, beginning of first and second therapy, sex ratio, duration of nephrosis before first cytotoxic therapy, interval between first and second cytotoxic therapy, number of relapses, cumulative doses of steroids and length of remission off steroids before second therapy were similar between groups. Four patients (36.4%, p<0.05) in group 1 remained in remission for a median 34.0 months, whereas only 1 patient (4.5%) in group 2 did for 53.0 months. Two patients in group 1 and 1 patient in group 2 became infrequent relapsers. Total number of nonrelapsers and infrequent relapsers was higher in group 1 (54.5%, p<0.05) than in group 2 (9.1%). Number of relapses and cumulative doses of steroids were reduced and length of remission off steroids was longer in group 1 than in group 2 (p<0.05). There was no difference between groups in frequency of side effects, and none had serious toxicity. However, the short period of follow-up in our study does not exclude the risk of azoospermia. CONCLUSIONS: Our data suggest a superior effect of chlorambucil over cyclophosphamide in steroid-dependent and cyclophosphamide-resistant children with MCNS. A future randomized controlled clinical trial is required to confirm our findings. PMID- 19809994 TI - Asymmetric dimethylarginine, vascular calcifications and parathyroid hormone serum levels in hemodialysis patients. AB - BACKGROUND: Asymmetric dimethylarginine (ADMA)is an endogenous amino acid similar to l-arginine and able to inhibit the enzyme endothelial nitric oxide synthase (eNOS). It is a factor of impaired nitric oxide (NO) synthesis. Serum levels of ADMA in chronic kidney disease (CKD) increase due to defective inactivation and excretion. High ADMA levels are associated with endothelial dysfunction and cardiovascular damage. A linkage between ADMA levels and vascular calcifications of CKD can therefore be hypothesized. This study explores also a possible relation between ADMA and parathyroid hormone (PTH) serum levels, which are known to be linked to increased rates of cardiovascular death. METHODS: The study was carried out in 79 patients on hemodialysis (HD), mean age 59.25 +/- 12 years. In all patients, serum ADMA, PTH, Ca, P, bone alkaline phosphatase (BALP), cholesterol and albumin were measured. In addition, the patients were subjected to multislice computed tomography for heart calcification evaluation. RESULTS: Correlation analysis of ADMA showed a significant relation with total and coronary calcium volumes, HD vintage, body mass index (BMI), cholesterol, serum albumin, PTH, natural logarithm of PTH (LnPTH) and BALP. Multiple regression analysis selected HD vintage, albumin and PTH as predictive variables for coronary calcium volume, while ADMA was excluded. With LnPTH as dependent variable, ADMA, serum calcium and BMI were predictive variables with R2 of 0.37. ADMA as dependent variable was also predicted by PTH, HD vintage, albumin and BMI. CONCLUSIONS: Despite the results of bivariate analysis showing a linkage between ADMA and cardiac and coronary calcifications, regression analysis showed only a spurious association. The strong positive correlation between ADMA and LnPTH, validated by the regression analysis, may suggesta link between ADMA and PTH-derived vascular damage. ADMA levels could be influenced by the severity of hyperparathyroidism and contribute to cardiovascular death linked to PTH of hemodialysis patients. PMID- 19809995 TI - Dimethylarginine levels and nutritional status in hemodialysis patients. AB - BACKGROUND: Asymmetric dimethylarginine (ADMA)and symmetric dimethylarginine (SDMA) originate from hydrolysis of methylated proteins, including dietary proteins, and are retained in end-stage renal disease(ESRD). This study aimed to detect the correlation of ADMA and SDMA to nutritional parameters in dialysis patients. METHODS: Before and after a single dialysis session, larginine, ADMA and SDMA plasma levels were measured in 38 hemodialysis patients by HPLC-tandem mass spectrometry. Biochemistry, protein intake, anthropometry and bioelectric impedance analysis were evaluated. RESULTS: Predialysis plasma levels of ADMA were higher than in normal controls (n=20) (1.14 +/- 0.27 mumol/Lvs. 0.56 +/- 0.09 mumol/L, p<0.001), as were SDMA levels(3.49 +/- 1.00 mumol/L vs. 0.44 +/- 0.13 mumol/L, p<0.001).On univariate analysis, predialysis ADMA levels were inversely related to BMI and albumin levels, whereas SDMA was directly related to nPNA, phase angle, prealbumin and creatinine serum levels. ADMA/SDMA ratio was inversely related to prealbumin and albumin, creatinine, urea and phosphorus serum levels, as wellas nPNA, but positively to C-reactive protein. On multiple regression analysis, serum albumin and BMI were the stronger predictors of ADMA, whereas prealbumin serum levels followed by dietary protein intake were the stronger predictors of SDMA. Prealbumin followed by C-reactive protein was predictive of the ADMA/SDMA molar ratio. CONCLUSIONS: Our results confirm that ADMA and SDMA levels are increased in ESRD patients and suggest that a link may exist with inflammation and nutritional status. High ADMA levels associated with reduced SDMA may be a predictive marker of malnutrition-inflammation atherosclerosis syndrome. PMID- 19809996 TI - Oxidative stress in elderly chronic renal failure patients: effects of renal replacement therapies on cell membrane fluidity. AB - BACKGROUND: Renal replacement therapies (RRTs) produce a partial loss of antioxidants and formation of reactive oxygen species (ROS), which are a major factor involved in alterations of plasma membrane fluidity and endothelial activation, but their role on plasma membrane fluidity in vivo is still unclear. We compared erythrocyte plasma membrane fluidity, ROS and total plasma antioxidant defenses (Lagtime) in aged patients with chronic renal failure (CRF) on conservative treatment, peritoneal dialysis (PD) and hemodialysis (HD) before (HD-pre) and after (HD-post) a treatment, to evaluate the role of different RRTs on oxidative stress and plasma membrane fluidity in aged patients. METHODS: We assessed erythrocyte plasma membrane fluidity, plasma lipid hydroperoxide levels and Lag-Time in 11 CRF patients on conservative treatment, 15 on PD, 12 on HD and 30 healthy controls. RESULTS: Hydroperoxides were higher in CRF, PD and HD-post, whereas Lag-time was significantly lower in PD, CRF and in HD-post. CRF, PD and HD-pre also had higher membrane fluidity (rsDPH), compared with HD-post and controls. CONCLUSION: These findings are in keeping with the hypothesis that the Lag-time decrease is due not only to the effect of the RRT but also to the uremic state, and that PD patients undergo a chronic, greater oxidative stress. Contrary to expectations, all patients showed greater erythrocyte membrane fluidity, which can be attributed to uremic toxicity. These observations reinforce the hypothesis that oxidative stress is an intrinsic component of this disease state and indeed is already present also in CRF not yet requiring RRT. PMID- 19809997 TI - Effect of weight loss after Roux-en-Y gastric bypass, on renal function and blood pressure in morbidly obese patients. AB - BACKGROUND: Morbid obesity (MO) is associated with increased renal plasma flow (RPL) and glomerular filtration rate (GFR). This type of obesity usually does not respond to medical treatment, with bariatric surgery being the current treatment of choice. The present study aimed to evaluate whether weight loss may reverse the glomerular hyperfiltration of MO patients. METHODS: This was a retrospective study of 140 patients submitted to Roux-en-Y gastric bypass (31.5% men, mean body mass index 46.17 +/- 5). Renal glomerular function and anthropometric and biochemical parameters were studied in patients before and 8 months after the surgery. GFR was determined by 24-hour urine samples. RESULTS: In the obese group, GFR before surgery was 148.7 +/- 35.2 ml/min. After the weight loss, GFR decreased to 113.8 +/- 31.7 ml/min (p<0.0001). Homeostasis model assessment insulin resistance and glycosylated hemoglobin values were higher in MO with hyperfiltration. Weight loss was associated with reduction in blood pressure and GFR. It was found that the variation in systolic and diastolic blood pressure was a predictor of change in GFR. CONCLUSION: This study shows that obesity-related glomerular hyperfiltration ameliorates after weight loss. The improvement in hyperfiltration may prevent the development. PMID- 19809998 TI - Alkalinization potentiates vascular calcium deposition in an uremic milieu. AB - BACKGROUND: Vascular calcification is a serious complication of chronic kidney disease. Acid-base balance is a relevant, albeit somewhat forgotten factor in the regulation of calcium deposition. Hemodialysis patients undergo repeated episodes of alkaline loading from the dialysate, resulting in prolonged alkalinization. We have hypothesized that extracellular alkalinization may promote vascular calcification. METHODS: Primary cultures of vascular smooth muscle cells were induced to calcify by the phosphate donor beta-glycerophosphate, in the presence of normal or uremic sera from hemodialysis patients and at different pH conditions. The influence of sodium bicarbonate supplementation for 2 months on aorta calcification was studied in 5/6 nephrectomized uremic rats. RESULTS: Uremic serum increased vascular smooth muscle cell calcification (twofold over nonuremic human serum at day 12, p<0.001). Alkalinization of the extracellular medium also increased vascular smooth muscle cell calcification. Increasing the extracellular pH from 7.42 to 7.53 resulted in a 2.5-fold increase in calcium accumulation at day 12 (p<0.05). In vivo, arterial calcification was significantly higher in alkalinized uremic animals (aorta calcification index, uremic + sodium bicarbonate, 164 +/- 57 units, vs. uremic + vehicle, 56 +/- 14 units; p<0.01). CONCLUSIONS: Alkalinization increases vascular calcification in cultured cells and uremic rats. These data may be used to optimize dialysate composition and the degree of alkalinization in calcification-prone individuals with advanced renal disease. PMID- 19809999 TI - Management of chronic kidney disease in Taiwan: room for quality improvement. AB - BACKGROUND: End-stage renal disease is epidemic in Taiwan. Although early intervention in chronic kidney disease (CKD) has proven to be effective in modifying its clinical course, little is known about the quality and trend of CKD care in Taiwan. METHODS: From a nationally representative database of 200,000 individuals, CKD patients were identified by diagnostic codes, and their medical interventions were extracted from service claims. The predictors for care delivery were analyzed in a logistic regression model. RESULTS: Of the 7,740 patients with CKD, only 54.8% had follow-up of renal function. Tests for anemia, calcium/ phosphorus and lipid profile were performed in 34.5%, 5.2% and 40.9% of patients, respectively. Most hypertensive CKD patients have used antihypertensive agents, but only 58.1% of them received angiotensinconverting enzyme inhibitors (ACEIs) or angiotensin receptor blockers (ARBs). Of CKD patients with diabetes, less than half had glycosylated hemoglobin measured, and only 49.7% received ACEIs/ARBs. We observed an encouraging trend in the use of ACEIs/ ARBs and in most aspects of laboratory monitoring. CONCLUSIONS: Management of CKD patients is suboptimal, and lack of awareness is common. Further work is needed to evaluate the impact of educational initiatives and multidisciplinary teams on outcomes in CKD patients. PMID- 19810000 TI - Effect of computer-assisted European Best Practice Guideline implementation on adherence and target attainment: ORAMA results. AB - BACKGROUND: The success of guidelines depends largely on effective implementation and uptake in clinical practice. The Optimal Renal Anemia Management Assessment (ORAMA) study investigated the impact of European Best Practice Guideline (EBPG) prompting on patient outcomes. METHODS: ORAMA was a prospective, international, multicenter, cluster-randomized study. Fifty-three centers in eight European countries enrolled patients with chronic kidney disease stage V receiving chronic dialysis. Patients were either anemic (hemoglobin [Hb] <11 g/dL [110 g/L]) or treated with erythropoiesis stimulating agents and/or iron supplementation. Centers were randomized to two groups, with or without access to a computerized clinical decision support (CDS) system. In a post hoc analysis,patients were further subdivided into adherence or non-adherence of investigators to the EBPG regardless of CDS usage. Primary endpoint was the proportion of patients with Hb >11 g/ dL (110 g/L), reflecting hematological targets in the revised EPBG. RESULTS: In this population of 599 dialysis patients, hematological targets did not differ in the presence or absence of a CDS system. There was a general shift towards improved patient distribution by Hb categories while the width of the distribution curves remained unchanged. The proportion of patients with Hb >11 g/dL (110 g/L) was higher among adherers (79% and 84% with or without CDS use, respectively) than non-adherers (59% and 57%, respectively). CONCLUSIONS: ORAMA is the first international study to show that adherence to EBPG improved attainment of anemia indices. The availability of a CDS system did not affect anemia management. PMID- 19810001 TI - Rosuvastatin does not affect intrarenal hemodynamics in patients with hypercholesterolemia. AB - BACKGROUND: It has been proposed that hyperlipidemia contributes to the progression of renal diseases, and conversely, that treatment with statins decreases the progressive decline of renal function. Increased glomerular pressure has been found to cause endothelial dysfunction of glomerular capillaries. However, the potential renoprotective effects of statin treatment have not been fully elucidated and so far no study has analyzed the effects of statin treatment on intrarenal hemodynamics. METHODS: Forty hypercholesterolemic patients were randomly assigned to receive rosuvastatin or placebo in a double blind crossover study. Renal plasma flow(RPF) and glomerular filtration rate (GFR) were determined by constant input clearance technique with p-aminohippurate (PAH) and inulin. Glomerular hydrostatic pressure (Pglom) and resistances of the afferent(RA) and efferent arterioles (RE) were calculated according to the model originally established by Gomez. RESULTS: RPF and GFR were similar after treatment with rosuvastatin and placebo. Neither Pglom (66.2 +/- 3.9vs. 66.4 +/- 5.1 mm Hg, p=0.861) nor RA (3,200 +/- 1,780 vs.3,188 +/- 1,870 dyn-s-cm-5, p=0.957) or RE (3,620 +/- 1,174vs. 3,490 +/- 1,272 dyn-s-cm-5, p=0.378) were affected by treatment with rosuvastatin, compared with placebo. CONCLUSIONS: The beneficial effects on renal function by statin treatment seem not to be mediated by changes in intrarenal hemodynamics in patients with hypercholesterolemia. PMID- 19810002 TI - Hemizygous Fabry disease associated with IgA nephropathy: a case report. AB - We present a 22-year-old male patient who showed both classical Fabry disease and IgA nephropathy. He had proteinuria (1.5 g/day), hypohidrosis and neuralgia with fever. Serum creatinine and blood urea nitrogen were 0.9 mg/dL and 11.4 mg/dL, respectively. Renal biopsy showed strikingly vacuolated podocytes and tubular epithelium cells. Myelin-like bodies were detected in podocytes, mesangial cells, endothelial cells and tubular epithelium cells by electron microscopy. On immunofluorescence microscopy, IgA and C3 deposits were detected in mesangial areas. From these results and a markedly low level of alpha-galactosidase A activity, this patient was diagnosed as having classical Fabry disease and IgA nephropathy. PMID- 19810003 TI - 1H and 13C NMR assignments of all three isomeric o-fluoronaphthaldehydes and three o-fluorophenanthrene aldehydes. AB - Three isomeric o-fluoronaphthaldehydes, 9-fluorophenanthrene, and three previously unreported o-fluorophenanthrene aldehydes were analyzed in detail by multiple NMR techniques to provide unambiguous assignment of structures and resonances. The six aldehydes serve as the key starting materials for novel chiral ligands used in highly enantioselective rhodium-catalyzed asymmetric hydrogenation reactions. PMID- 19810004 TI - High-performance affinity chromatography method for identification of L-arginine interacting factors using magnetic nanobeads. AB - L-Arginine exhibits a wide range of biological activities through a complex and highly regulated set of pathways that remain incompletely understood at both the whole-body and the cellular levels. The aim of this study is to develop and validate effective purification system for L-arginine interacting factors (AIFs). We have recently developed novel magnetic nanobeads (FG beads) composed of magnetite particles/glycidyl methacrylate (GMA)-styrene copolymer/covered GMA. These nanobeads have shown higher performance compared with commercially available magnetic beads in terms of purification efficiency. In this study, we have newly developed L-arginine methyl ester (L-AME)-immobilized beads by conjugating L-AME to the surface of these nanobeads. Firstly, we showed that inducible nitric oxide synthase, which binds and uses L-arginine as a substrate, specifically bound to L-AME-immobilized beads. Secondly, we newly identified phosphofructokinase, RuvB-like 1 and RuvB-like 2 as AIFs from crude extracts of HeLa cells using this affinity chromatographic system. The data presented here demonstrate that L-AME-immobilized beads are effective tool for purification of AIFs directly from crude cell extracts. We expect that the present method can be used to purify AIFs from various types of cells. PMID- 19810005 TI - A sensitive non-aqueous capillary electrophoresis-mass spectrometric method for multiresidue analyses of beta-agonists in pork. AB - Non-aqueous capillary electrophoresis-mass spectrometry (NACE-MS) was developed for trace analyses of beta-agonists (i.e. clenbuterol, salbutamol and terbutaline) in pork. The NACE was in 18 mM ammonium acetate in methanol acetonitrile-glacial acetic acid (66 : 33 : 1, v/v/v) using a voltage of 28 kV. The hyphenation of CE with a time-of-flight MS was performed by electrospray ionization interface employing 5 mM ammonium acetate in methanol-water (80 : 20, v/v) as the sheath liquid at a flow rate of 2 microL/min. Method sensitivity was enhanced by a co-injection technique (combination of hydrodynamic and electrokinetic injection) using a pressure of 50 mbar and a voltage of 10 kV for 12 s. The method was validated in comparison with HPLC-MS-MS. The NACE-MS procedure provided excellent detection limits of 0.3 ppb for all analytes. Method linearity was good (r(2) > 0.999, in a range of 0.8-1000 ppb for all analytes). Precision showed %RSDs of <17.7%. Sample pre-treatment was carried out by solid phase extraction using mixed mode reversed phase/cation exchange cartridges yielding recoveries between 69 and 80%. The NACE-MS could be successfully used for the analysis of beta-agonists in pork samples and results showed no statistical differences from the values reported by the Ministry of Public Health, Thailand using HPLC-MS-MS method. PMID- 19810006 TI - Determination of dopamine, serotonin, and their metabolites in pediatric cerebrospinal fluid by isocratic high performance liquid chromatography coupled with electrochemical detection. AB - A method to rapidly measure dopamine (DA), dihydroxyindolphenylacetic acid, homovanillic acid, serotonin (5-HT) and 5-hydroxyindoleacetic acid concentrations in cerebrospinal fluid (CSF) has not yet been reported. A rapid, sensitive, and specific HPLC method was therefore developed using electrochemical detection. CSF was mixed with an antioxidant solution prior to freezing to prevent neurotransmitter degradation. Separation of the five analytes was obtained on an ESA MD-150 x 3.2 mm column with a flow rate of 0.37 mL/min and an acetonitrile aqueous (5 : 95, v/v) mobile phase with 75 mM monobasic sodium phosphate buffer, 0.5 mM EDTA, 0.81 mM sodium octylsulfonate and 5% tetrahydrofuran. The optimal electrical potential settings were: guard cell +325 mV, E1 -100 mV and E2 +300 mV. Within-day and between-day precisions were <10% for all analytes and accuracies ranged from 91.0 to 106.7%. DA, 5-HT, and their metabolites were stable in CSF with antioxidant solution at 4 degrees C for 8 h in the autoinjector. This method was used to measure neurotransmitters in CSF obtained from children enrolled on an institutional medulloblastoma treatment protocol. PMID- 19810007 TI - Bioanalysis of pentoxifylline and related metabolites in plasma samples through LC-MS/MS. AB - Analytical aspects related to the assay of pentoxifylline (PTX), lisofylline (M1) and carboxypropyl dimethylxanthine (M5) metabolites are discussed through comparison of two alternative analytical methods based on liquid chromatography separation and atmospheric pressure electrospray ionization tandem mass spectrometry detection. One method is based on a 'pure' reversed-phase liquid chromatography mechanism, while the second one uses the additional polar interactions with embedded amide spacers linking octadecyl moieties to the silicagel surface (C-18 Aqua stationary phase). In both cases, elution is isocratic. Both methods are equally selective and allows separation of unknowns (four species associated to PTX, two species associated to M1) detected through specific mass transitions of the parent compounds and owning respective structural confirmation. Plasma concentration-time patterns of these compounds follow typical metabolic profiles. It has been advanced that in-vivo formation of conjugates of PTX and M1 is possible, such compounds being cleaved back to the parent ones within the ion source. The first method was associated with a sample preparation procedure based on plasma protein precipitation by strong organic acid addition. The second method used protein precipitation by addition of a water miscible organic solvent. Both analytical methods were fully validated and used to assess bioequivalence between a prolonged release generic formulation and the reference product, under multidose and single dose approaches. PMID- 19810008 TI - Highly sensitive determination of Schisandrin and Schisandrin B in plasma of rats after administration of Wurenchun (Fructus Schisandrae Chinensis Extracts) preparations by LC-ESI-MS/MS. AB - A sensitive and specific method based on liquid chromatography-tandem mass spectrometry using electrospray ionization (LC-ESI-MS/MS) has been developed for the determination of Schisandrin and Schisandrin B in rat plasma. A 100 microL plasma sample was extracted by methyl tert-butyl ether after spiking the samples with nimodipine (internal standard) and performed on an XTerra(R)MS-C(18) column (150 mm x 2.1 mm, 3.5 mum) with the mobile phase of acetonitrile-water-formic acid (80:20:0.2, v/v) at a flow rate of 0.2 mL/min in a run time of 8.5 min. The lower limit of quantification of the method was 40 ng/mL for Schisandrin and 20 ng/mL for Schisandrin B. The method showed reproducibility with intra-day and inter-day precision of less than 13.8% RSD, as well as accuracy, with inter- and intra-assay accuracies between 93.5 and 107.2%. Finally, the LC-ESI-MS/MS method was successfully applied to study the pharmacokinetics of Schisandrin and Schisandrin B in rats after administration of Wurenchun commercial formulations to rats. PMID- 19810009 TI - Liquid chromatography tandem mass spectrometry method for determination of bisoprolol in human plasma using d5-bisoprolol as the internal standard. AB - A simple, reliable and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) protocol was developed and validated for quantification of bisoprolol in human plasma. The sample was pretreated with a simple procedure of protein precipitation and an isotope-labeled d5-bisoprolol was used as internal standard. The chromatographic separation was performed on a Capcell Pak C(18) MG III column (100 mm x 2.0 mm, 5 microm). The protonated ion of the analyte was detected in positive ionization by multiple reaction monitoring mode. The mass transition pairs of m/z 326.3 --> 116.3 and m/z 331.3 --> 121.3 were used to detect bisoprolol and the internal standard, respectively. Linearity, accuracy, precision, recovery, matrix effect, dilution test and stability were evaluated during method validation over the range of 0.5-100 ng/mL. The validated method was successfully applied to analyze human plasma samples in a bisoprolol bioavailability study. PMID- 19810010 TI - Executive functioning in Alzheimer's disease and vascular dementia. AB - OBJECTIVE: To compare performance of patients with mild-moderate Alzheimer's disease (AD) and vascular dementia (VaD) on tests of executive functioning and working memory. METHODS: Patients with AD (n = 76) and VaD (n = 46) were recruited from a memory clinic along with dementia free participants (n = 28). They underwent specific tests of working memory from the Cognitive Drug Research (CDR) battery and pen and paper tests of executive function including CLOX 1 & 2, EXIT25 and a test of verbal fluency (COWAT). All patients had a CT brain scan which was independently scored for white matter change/ischaemia. RESULTS: The AD and VaD groups were significantly impaired on all measures of working memory and executive functioning compared to the disease free group. There were no significant differences between the AD and VaD groups on any measure. Z-scores confirmed the pattern of impairment in executive functioning and working memory was largely equivalent in both patient groups. Small to moderate correlations were seen between the MMSE and the neurocognitive scores in both patient groups and the pattern of correlations was also very similar in both patient groups. CONCLUSIONS: This study demonstrates sizeable executive functioning and working memory impairments in patients with mild-moderate AD and VaD but no significant differences between the disease groups. PMID- 19810012 TI - Severe iron overload in Blackfan-Diamond anemia: a case-control study. AB - Chronic iron overload is a serious complication in transfusion-dependent patients. Few studies have addressed this issue in Diamond-Blackfan anemia (DBA). We describe a retrospective analysis of iron overload, and its related complications in 31 transfusion-dependent Italian DBA patients whose records included one or more evaluation of liver iron concentration (LIC) by means of noninvasive magnetic liver susceptometry with a superconductive quantum interference device (SQUID). This cohort is also matched with a group of transfusion-dependent beta-thalassemia major patients to look for differences. A severe iron overload was observed in 54% patients, especially among those inadequately chelated. The DBA patients displayed a significantly higher LIC than the regularly chelated beta-thalassemics. This difference may have been attributable to nonoptimal chelation (late onset, type, dose, prescription, and compliance), or an unknown biological mechanism that lead to an early severe iron overload. We therefore suggest that all transfusion patients should have an accurate record of their iron intake, a regular monitoring of iron overload, in order to start chelation when a critical transfusion load is reached, and to test the efficacy/compliance of chelation treatment. Physicians taking care of transfusion-dependent DBA patients must be concerned about the frequent and early complications such as cardiac toxicity. Am. J. Hematol., 2009. (c) 2009 Wiley Liss, Inc. PMID- 19810011 TI - Production of beta-globin and adult hemoglobin following G418 treatment of erythroid precursor cells from homozygous beta(0)39 thalassemia patients. AB - In several types of thalassemia (including beta(0)39-thalassemia), stop codon mutations lead to premature translation termination and to mRNA destabilization through nonsense-mediated decay. Drugs (for instance aminoglycosides) can be designed to suppress premature termination, inducing a ribosomal readthrough. These findings have introduced new hopes for the development of a pharmacologic approach to the cure of this disease. However, the effects of aminoglycosides on globin mRNA carrying beta-thalassemia stop mutations have not yet been investigated. In this study, we have used a lentiviral construct containing the beta(0)39-thalassemia globin gene under control of the beta-globin promoter and a LCR cassette. We demonstrated by fluorescence-activated cell sorting (FACS) analysis the production of beta-globin by K562 cell clones expressing the beta(0)39-thalassemia globin gene and treated with G418. More importantly, after FACS and high-performance liquid chromatography (HPLC) analyses, erythroid precursor cells from beta(0)39-thalassemia patients were demonstrated to be able to produce beta-globin and adult hemoglobin after treatment with G418. This study strongly suggests that ribosomal readthrough should be considered a strategy for developing experimental strategies for the treatment of beta(0)-thalassemia caused by stop codon mutations. Am. J. Hematol., 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19810013 TI - Length distribution of single-walled carbon nanotubes in aqueous suspension measured by electrospray differential mobility analysis. AB - The first characterization of the length distribution of single-walled carbon nanotubes (SWCNT) dispersed in a liquid by electrospray differential mobility analysis (ES-DMA) is presented. Although an understanding of geometric properties of SWCNTs, including length, diameter, aspect ratio, and chirality, is essential for commercial applications, rapid characterization of nanotube length distributions remains challenging. Here the use of ES-DMA to obtain length distributions of DNA-wrapped SWCNTs dispersed in aqueous solutions is demonstrated. Lengths measured by ES-DMA compare favorably with those obtained from multiangle light scattering, dynamic light scattering, field flow fractionation with UV/vis detection, and atomic force microscopy, validating ES DMA as a technique to measure SWCNTs of <250 nm in length. The nanotubes are previously purified and dispersed by wrapping with oligomeric DNA in aqueous solution and centrifuging to remove bundles and amorphous carbon. These dispersions are particularly attractive due to their amenability to bulk processing, ease of storage, high concentration, compatibility with biological and high-throughput manufacturing environments, and for their potential applications ranging from electronics and hydrogen-storage vessels to anticancer agents. PMID- 19810014 TI - Determination of pyridoxal-5'-phosphate (PLP)-bonding sites in proteins: a peptide mass fingerprinting approach based on diagnostic tandem mass spectral features of PLP-modified peptides. AB - Peptides modified by pyridoxal-5'-phosphate (PLP), linked to a lysine residue via reductive amination, exhibit distinct spectral characteristics in the collision induced dissociation (CID) tandem mass (MS/MS) spectra that are described here. The MS/MS spectra typically display two dominant peaks whose m/z values correspond to neutral losses of [H3PO4] (-98 Da) and the PLP moiety as [C8H10NO5P] (-231 Da) from the precursor peptide ion, respectively. Few other peaks are observed. Recognition of this distinct fragmentation behavior is imperative since determining sequences and sites of modifications relies on the formation of amide backbone cleavage products for subsequent interpretation via proteome database searching. Additionally, PLP-modified peptides exhibit suppressed precursor ionization efficiency which diminishes their detection in complex mixtures. Presented here is a protocol which describes an enrichment strategy for PLP-modified peptides combined with neutral loss screening and peptide mass fingerprinting to map the PLP-bonding site in a known PLP-dependent protein. This approach represents an efficient alternative to site-directed mutagenesis which has been the traditional method used for PLP-bonding site localization in proteins. PMID- 19810015 TI - An electropneumatic-heated nebulizer for enhancing spray ionization in PhotoSpray atmospheric pressure photoionization sources for liquid chromatography/mass spectrometry. AB - We introduce a novel electropneumatic-heated nebulizer (EPn-HN), incorporating an electrified internal pneumatic nebulizer, to enhance the yield of sprayed ions from PhotoSpray atmospheric pressure photoionization (APPI) sources for liquid chromatography/mass spectrometry (LC/MS). Spray ionization from the pneumatic heated nebulizers used in APPI sources provides a supplemental, complementary ionization method to be used for involatile and thermally labile compounds, otherwise intractable to APPI. Details of the construction and operation of the EPn-HN device are provided. The performance of the EPn-HN is demonstrated using two model compounds: substance P, a peptide used as a standard in studies of ion fragmentation mechanisms, and aztreonam, a thermally labile antibiotic. At the optimum voltage for spray ionization, improvements in sensitivity of two orders of magnitude are obtained relative to when the sprayer is grounded, the conventional case. Since both substance P and aztreonam cannot be detected using the APPI method alone, the results demonstrate how spray ionization from the EPn HN may be used to extend the range of compounds amenable to PhotoSpray sources. PMID- 19810018 TI - Naringin inhibits matrix metalloproteinase-9 expression and AKT phosphorylation in tumor necrosis factor-alpha-induced vascular smooth muscle cells. AB - Citrus fruits are high in naringin, which has a beneficial effect on cardiovascular diseases. However, the matrix metalloproteinase-9 (MMP-9) regulation involved in cell migration and invasion remains to be identified. Naringin inhibited tumor necrosis factor-alpha (TNF-alpha)-induced expression of MMP-9, under 10-25 microM concentration conditions in vascular smooth muscle cells (VSMC). The TNF-alpha-induced invasion and migration of VSMC were inhibited by naringin. Furthermore, naringin suppressed TNF-alpha-mediated release of interleukin-6 and -8 (IL-6 and IL-8). However, naringin (10-25 microM) treatment of VSMC in the presence of TNF-alpha did not affect cell growth and apoptosis. In additional experiments, naringin reduced the transcriptional activity of activator protein-1 and nuclear factor kappaB (NF-kappaB), which are two important nuclear transcription factors that are involved in MMP-9 expression. Also, naringin treatment blocked PI3K/AKT/mTOR/p70S6K pathway in TNF-alpha induced VSMC. Treatment of aglycone naringenin (10-25 microM) had same effect on the levels of MMP-9 expression, invasion, migration, and AKT phosphorylation in TNF-alpha-induced VSMC, compared with naringin treatment. These results suggest that naringin represses PI3K/AKT/mTOR/p70S6K pathway, invasion and migration, and subsequently suppresses MMP-9 expression through the transcription factors NF kappaB and activator protein-1 in TNF-alpha-induced VSMC. These novel findings provide a theoretical basis for the preventive use of naringin for atherosclerosis disease. PMID- 19810019 TI - Influence of thermal processing on IgE reactivity to lentil and chickpea proteins. AB - In the last years, legume proteins are gaining importance as food ingredients because of their nutraceutical properties. However, legumes are also considered relevant in the development of food allergies through ingestion. Peanuts and soybeans are important food allergens in Western countries, while lentil and chickpea allergy are more relevant in the Mediterranean area. Information about the effects of thermal-processing procedures at various temperatures and conditions is scarce; therefore, the effect of these procedures on legume allergenic properties is not defined so far. The SDS-PAGE and IgE-immunoblotting patterns of chickpeas and lentils were analyzed before and after boiling (up to 60 min) and autoclaving (1.2 and 2.6 atm, up to 30 min). The results indicated that some of these treatments reduce IgE binding to lentil and chickpea, the most important being harsh autoclaving. However, several extremely resistant immunoreactive proteins still remained in these legumes even after this extreme treatment. PMID- 19810020 TI - Hollow fibre-supported liquid membrane extraction and LC-MS/MS detection for the analysis of heterocyclic amines in urine samples. AB - Heterocyclic amines (HCAs) are potent mutagens/carcinogens to which humans are frequently exposed through the consumption of cooked meat and fish food. The effect of normal intake of HCAs and their role in the aetiology of human cancer is unknown. To some extent, limitations of the existing analytical methods in monitoring the low levels of HCAs in biological samples have hindered obtaining conclusive results. In this study, a method for the analysis of HCAs in human urine has been studied to detect HCAs and metabolites at levels resulting from consumption of food cooked at ordinary conditions. The analytical method consisted of extraction and clean-up by the novel technique liquid-phase microextraction combined with LC-MS/MS. The effect of pH during the extraction and hydrolysis step was examined. High sensitivity was achieved when the extraction was performed in raw urine adjusted to pH 5.5, 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine being detected from 2 pg/g urine, levels comparable with a normal exposure. Good reproducibility and repeatability was obtained for 2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine and 2-amino-3,8-dimethylimidazo[4,5 f]quinoxaline, below 9% using isotopic dilution. The performance of the method on 9H-pyrido[3,4-b]indole, 2-amino-1-methyl-6-(4'-hydroxyphenyl)imidazo[4,5 b]pyridine and 2-amino-1-methyl-6-(5-hydroxy)phenylimidazo[4,5-b]pyridine was also studied. PMID- 19810021 TI - Four selenoproteins, protein biosynthesis, and Wnt signalling are particularly sensitive to limited selenium intake in mouse colon. AB - Selenium is an essential micronutrient. Its recommended daily allowance is not attained by a significant proportion of the population in many countries and its intake has been suggested to affect colorectal carcinogenesis. Therefore, microarrays were used to determine how both selenoprotein and global gene expression patterns in the mouse colon were affected by marginal selenium deficiency comparable to variations in human dietary intakes. Two groups of 12 mice each were fed a selenium-deficient (0.086 mg Se/kg) or a selenium-adequate (0.15 mg Se/kg) diet. After 6 wk, plasma selenium level, liver, and colon glutathione peroxidase (GPx) activity in the deficient group was 12, 34, and 50%, respectively, of that of the adequate group. Differential gene expression was analysed with mouse 44K whole genome microarrays. Pathway analysis by GenMAPP identified the protein biosynthesis pathway as most significantly affected, followed by inflammation, Delta-Notch and Wnt pathways. Selected gene expression changes were confirmed by quantitative real-time PCR. GPx1 and the selenoproteins W, H, and M, responded significantly to selenium intake making them candidates as biomarkers for selenium status. Thus, feeding a marginal selenium-deficient diet resulted in distinct changes in global gene expression in the mouse colon. Modulation of cancer-related pathways may contribute to the higher susceptibility to colon carcinogenesis in low selenium status. PMID- 19810022 TI - Variation in the human lipidome associated with coffee consumption as revealed by quantitative targeted metabolomics. AB - The effect of coffee consumption on human health is still discussed controversially. Here, we report results from a metabolomics study of coffee consumption, where we measured 363 metabolites in blood serum of 284 male participants of the Cooperative Health Research in the Region of Augsburg study population, aged between 55 and 79 years. A statistical analysis of the association of metabolite concentrations and the number of cups of coffee consumed per day showed that coffee intake is positively associated with two classes of sphingomyelins, one containing a hydroxy-group (SM(OH)) and the other having an additional carboxy-group (SM(OH,COOH)). In contrast, long- and medium chain acylcarnitines were found to decrease with increasing coffee consumption. It is noteworthy that the concentration of total cholesterol also rises with an increased coffee intake in this study group. The association observed here between these hydroxylated and carboxylated sphingolipid species and coffee intake may be induced by changes in the cholesterol levels. Alternatively, these molecules may act as scavengers of oxidative species, which decrease with higher coffee intake. In summary, we demonstrate strong positive associations between coffee consumption and two classes of sphingomyelins and a negative association between coffee consumption and long- and medium-chain acylcarnitines. PMID- 19810023 TI - Acoustic analysis of voice in cochlear implant recipients with post-meningitic hearing loss. AB - The objective of this study was to evaluate the critical time period between the onset of sensorineural hearing loss and cochlear implantation with respect to normal voice production in children with post-meningitic hearing loss. Acoustic measures of voice production were obtained from ten paediatric cochlear implant recipients with post-meningitic hearing loss. Acoustic measures were obtained utilising the Multi-Dimensional Voice Program and Computerized Speech Laboratory (Kay Elemetrics Corp.). Measures were based on sustained phonation of the vowel /a/. Acoustic parameters included fundamental frequency, short- and long-term frequency perturbation, and short- and long-term amplitude perturbation. Measures of fundamental frequency and short-term frequency and amplitude perturbation were comparable to values of children with normal hearing. Long-term control of frequency was within normal limits for subjects with a period of auditory deprivation of less than four months. Measures of long-term amplitude perturbation were normal for all patients except those with cochlear ossification. Early restoration of auditory feedback with cochlear implantation, the absence of cochlear ossification, residual aided hearing following meningitis, and auditory-verbal therapy were identified as factors in preserving the long-term control of frequency and amplitude in the setting of post meningitic hearing loss. PMID- 19810024 TI - Comparisons of multi-marker association methods to detect association between a candidate region and disease. AB - The joint use of information from multiple markers may be more effective to reveal association between a genomic region and a trait than single marker analysis. In this article, we compare the performance of seven multi-marker methods. These methods include (1) single marker analysis (either the best scoring single nucleotide polymorphism in a candidate region or a combined test based on Fisher's method); (2) fixed effects regression models where the predictors are either the observed genotypes in the region, principal components that explain a proportion of the genetic variation, or predictors based on Fourier transformation for the genotypes; and (3) variance components analysis. In our simulation studies, we consider genetic models where the association is due to one, two, or three markers, and the disease-causing markers have varying allele frequencies. We use information from either all the markers in a region or information only from tagging markers. Our simulation results suggest that when there is one disease-causing variant, the best-scoring marker method is preferred whereas the variance components method and the principal components method work well for more common disease-causing variants. When there is more than one disease-causing variant, the principal components method seems to perform well over all the scenarios studied. When these methods are applied to analyze associations between all the markers in or near a gene and disease status for an inflammatory bowel disease data set, the analysis based on the principal components method leads to biologically more consistent discoveries than other methods. PMID- 19810025 TI - An evaluation of statistical approaches to rare variant analysis in genetic association studies. AB - Genome-wide association (GWA) studies have proved to be extremely successful in identifying novel common polymorphisms contributing effects to the genetic component underlying complex traits. Nevertheless, one source of, as yet, undiscovered genetic determinants of complex traits are those mediated through the effects of rare variants. With the increasing availability of large-scale re sequencing data for rare variant discovery, we have developed a novel statistical method for the detection of complex trait associations with these loci, based on searching for accumulations of minor alleles within the same functional unit. We have undertaken simulations to evaluate strategies for the identification of rare variant associations in population-based genetic studies when data are available from re-sequencing discovery efforts or from commercially available GWA chips. Our results demonstrate that methods based on accumulations of rare variants discovered through re-sequencing offer substantially greater power than conventional analysis of GWA data, and thus provide an exciting opportunity for future discovery of genetic determinants of complex traits. PMID- 19810026 TI - Use of coated capillaries for the electrophoretic separation of stereoisomers of a growth hormone secretagogue. AB - The diastereoisomeric separation of peptidomimetics of hexarelin, a strong growth hormone secretagogue, in CE has been studied. Highly sulfated-gamma-CD was found to be an appropriate selector for the separation of the stereoisomers. However, non-repeatable analyses were obtained on bare fused silica capillary due to the progressive adsorption of the analytes on the capillary wall. Two types of polyelectrolyte coating agents were tested to prevent this phenomenon. Coating with neutral polyethylene oxide was found to be efficient but resulted in a very long analysis time (about 40 min). Coating with cationic poly(diallyldimethylammonium) chloride was found both to prevent analyte adsorption, reduce analysis time and alter separation selectivity. EOF measurement revealed that the highly sulfated-gamma-CDs were strongly adsorbed on the poly(diallyldimethylammonium) chloride coating surface yielding a stable strong cathodic EOF, which considerably reduced analysis time (about 12 min). Very good repeatability of analysis was obtained (RSD(migration time)<1%). PMID- 19810027 TI - SNaPshot minisequencing to resolve mitochondrial macro-haplogroups found in Africa. AB - African mitochondrial DNA (mtDNA) haplogroups are divided into seven macro haplogroups (L0'1'2'3'4'5'6), while the rest of the world's lineages are classified as subgroups of macro-haplogroups M, N and R. The most common approach to characterizing mtDNA variation is the sequencing of hypervariable segments I and II of the non-coding control region of the molecule. Given the higher mutation rate within the control region compared with the coding regions of the molecule, recurrent mutations in the former can sometimes hide possible phylogenetic structure. The incorporation of haplogroup-defining coding region mutations has helped in overcoming this limitation. By judiciously selecting 14 coding region SNPs and incorporating them into a multiplex minisequencing assay we were able to resolve mtDNA sequences from some sub-Saharan African populations into ten macro-haplogroups (L0-L6, M, N and R). We tested the efficacy of the panel by screening 699 individuals, consisting mostly of Khoe-San, Bantu speakers and individuals with mixed ancestries (Coloreds) and found no inconsistencies compared with hypervariable segment sequencing results. The panel provided a fast and efficient means of classifying mtDNA into the ten mitochondrial macro haplogroups and provided a reliable screening to distinguish African from non African-derived mtDNA lineages. PMID- 19810028 TI - Dielectrophoretic manipulation and separation of microparticles using curved microelectrodes. AB - This paper presents the development and experimental analysis of a dielectrophoresis (DEP) system, which is used for the manipulation and separation of microparticles in liquid flow. The system is composed of arrays of microelectrodes integrated to a microchannel. Novel curved microelectrodes are symmetrically placed with respect to the centre of the microchannel with a minimum gap of 40 microm. Computational fluid dynamics method is utilised to characterise the DEP field and predict the dynamics of particles. The performance of the system is assessed with microspheres of 1, 5 and 12 microm diameters. When a high-frequency potential is applied to microelectrodes a spatially varying electric field is induced in the microchannel, which creates the DEP force. Negative-DEP behaviour is observed with particles being repelled from the microelectrodes. The particles of different dimensions experience different DEP forces and thus settle to separate equilibrium zones across the microchannel. Experiments demonstrate the capability of the system as a field flow fraction tool for sorting microparticles according to their dimensions and dielectric properties. PMID- 19810029 TI - Electrophoretic exclusion for the selective transport of small molecules. AB - A novel method capable of differentiating and concentrating small molecules in bulk solution termed "electrophoretic exclusion" is described and experimentally investigated. In this technique, the hydrodynamic flow of the system is countered by the electrophoretic velocity to prevent a species from entering into the channel. The separation can be controlled by changing the flow rate or applied electric field in order to exclude certain species selectively while allowing others to pass through the capillary. Proof of principle studies employed a flow injection regime of the method and examined the exclusion of Methyl Violet dye in the presence of a neutral species. Methyl Violet was concentrated almost 40 times the background concentration in 30 s using 6 kV. Additionally, a threshold voltage necessary for exclusion was determined. The establishment of a threshold voltage enabled the differentiation of two similar cationic species: Methyl Green and Neutral Red. PMID- 19810030 TI - Simultaneous extraction of nucleic acids and proteins from tissue specimens by ultracentrifugation: A protocol using the high-salt protein fraction for quantitative proteome analysis. AB - Comprehensive molecular profiling of human tumor tissue specimens at the DNA, mRNA and protein level is often obstructed by a limited amount of available material. Homogenization of frozen tissue samples in guanidine isothiocyanate followed by ultracentrifugation over cesium chloride allows the simultaneous extraction of high-molecular weight DNA and RNA. Here, we present a protocol for quantitative proteome analysis using the high-salt protein fraction obtained as supernatant after ultracentrifugation for nucleic acid extraction. We applied this method to extracts from primary human brain tumors and demonstrate its successful application for protein expression profiling in these tumors using 2-D DIGE, MS and Western blotting. PMID- 19810031 TI - Proteomic analysis of Arabidopsis halleri shoots in response to the heavy metals cadmium and zinc and rhizosphere microorganisms. AB - Arabidopsis halleri has the rare ability to colonize heavy metal-polluted sites and is an emerging model for research on adaptation and metal hyperaccumulation. The aim of this study was to analyze the effect of plant-microbe interaction on the accumulation of cadmium (Cd) and zinc (Zn) in shoots of an ecotype of A. halleri grown in heavy metal-contaminated soil and to compare the shoot proteome of plants grown solely in the presence of Cd and Zn or in the presence of these two metals and the autochthonous soil rhizosphere-derived microorganisms. The results of this analysis emphasized the role of plant-microbe interaction in shoot metal accumulation. Differences in protein expression pattern, identified by a proteomic approach involving 2-DE and MS, indicated a general upregulation of photosynthesis-related proteins in plants exposed to metals and to metals plus microorganisms, suggesting that metal accumulation in shoots is an energy demanding process. The analysis also showed that proteins involved in plant defense mechanisms were downregulated indicating that heavy metals accumulation in leaves supplies a protection system and highlights a cross-talk between heavy metal signaling and defense signaling. PMID- 19810032 TI - Proteomic analysis of the secretome of human umbilical vein endothelial cells using a combination of free-flow electrophoresis and nanoflow LC-MS/MS. AB - Human umbilical vein endothelial cells are the most widely used in vitro model for endothelial cells. Their secreted proteins, however, have not been comprehensively analysed so far. In this study, we accomplished to map the secretome of human umbilical vein endothelial cells by combining free-flow electrophoresis with nanoflow LC-MS/MS. This comprehensive analysis provides a basis for future comparative studies of protein secretion by endothelial cells in response to cardiovascular risk factors and is available on our website http://www.vascular-proteomics.com. PMID- 19810033 TI - ExoCarta: A compendium of exosomal proteins and RNA. AB - Exosomes, membrane microvesicles (40-100 nm) secreted by most cell types, can be isolated in several ways while characterizing them is heavily based on electron microscopy and, most importantly, the identification of exosome marker proteins. Researchers rely on the identification of certain exosomal marker proteins including Alix, CD9 and CD63 to confirm the presence of exosomes in their preparations. An evolutionary-conserved set of protein molecules have been identified in most exosomes studied to date. However, with the complexity of tissue/cell type-specific proteins being incorporated in the exosomes, some of these so-called exosomal markers are not always present in all the exosomes. The presence of tissue/cell type-specific proteins in exosomes allows researchers to isolate them using immunoaffinity capture methods. A compendium for exosomal proteomes will aid researchers in identifying proteins that were more commonly found in various exosomes (exosome markers) and those that are specific to certain tissue/cell type-derived exosomes. Here, we describe ExoCarta, a compendium for proteins and RNA molecules identified in exosomes. ExoCarta is first of its kind and the resource is freely available to the scientific community through the web (http://exocarta.ludwig.edu.au). We believe that this community resource will be of great biological importance for any future exosome analyses. PMID- 19810034 TI - Can ultrasonic energy efficiently speed (18)O-labeling of proteins? AB - We report in this work on the robustness of ultrasonic energy as a tool to speed the isotopic labeling of proteins using the (18)O-decoupling procedure. The first part of the decoupling procedure, comprising protein denaturation, reduction, alkylation and digestion, is done in 8 min under the effects of an ultrasonic field whilst the second part, the isotopic labeling, was assayed with and without the use of ultrasonic energy. Our results clearly demonstrate that the (18)O isotopic labeling in a decoupling procedure cannot be accelerated using an ultrasonic field. PMID- 19810035 TI - Combining selected reaction monitoring with discovery proteomics in limited biological samples. AB - Simultaneous quantification of multiple proteins by selected reaction monitoring (SRM) has several applications in cell signaling studies including embryo proteomics. However, concerns have recently been raised over the specificity of SRM assays due to possible ion redundancy and/or sequence similarity of selected peptide with multiple non-related proteins. In this Viewpoint article, we discuss some simple measures that can increase our confidence in the accuracy of SRM scans used in proteomic experiments. At least in embryonic samples from porcine species, these measures were found to be useful in validating MS-identified differentially expressed proteins. Among the nine proteins analyzed by SRM assay, all the proteins that were found to be up- or down-regulated in MS experiment were also faithfully up- or down-regulated in SRM assay. PMID- 19810036 TI - Efficient inhibition of human cytomegalovirus UL122 gene expression in cell by small interfering RNAs. AB - In order to develop a gene therapy to human cytomegalovirus (HCMV), RNA interference (RNAi) was employed to inhibit the expression of HCMV UL122 gene in vitro. Recombinant vector pUL122-EGFP, which expressed UL122-EGFP fusion protein, and recombinant vectors psi122-1, psi122-2 and psi122-3, which expressed small interfering RNAs (siRNAs) targeted to UL122 were contransfected into AD293 cells. The fluorescence signal of pUL122-EGFP was greatly suppressed by psi122-1 and psi122-2, with an inhibitory rate of 82.0% +/- 1.0% and 79.5% +/- 2.5%, respectively. The mRNA of pUL122-EGFP of the cells transfected with psi122-1 and psi122-2 was decreased 97.3% +/- 0.6% and 98.0% +/- 0.1%, respectively. Vector psi122-3 showed a slightly low suppression rate. Therefore, it may be concluded that plasmids encoding siRNAs targeted to UL122 is able to in vitro reduce markedly the expression of UL122-EGFP. And it is very likely that the psi122-1 and psi122-2 are potentially efficacious siRNAs in the gene therapy of HCMV infection in vivo, in which further investigations are required. This study is expected to greatly facilitate the use of the RNAi technology for the anti-HCMV studies. PMID- 19810037 TI - Genotypic characterization of non starter lactic acid bacteria involved in the ripening of artisanal Bitto PDO cheese. AB - Bitto of Valchiavenna, an artisanal Italian cheese produced without the addition of any starter cultures, has been attributed a protected designation origin (PDO) cheese, but the strain composition of the natural microbial population colonizing this traditional dairy product is still unknown. To obtain preliminary information on the non starter lactic acid bacteria involved in its ripening, a total of 136 NSLAB isolates, randomly selected from MRS and M17 agar plates, were collected from three different cheese samples after 120 days of ripening. The new isolates were identified by combining PCR 16S-23S rDNA spacer analyses, partial 16S rRNA gene sequencing, species-specific probes and colony hybridization. Eighty-two isolates, representing 60% of the total strains selected, were homofermentative cocci: 83% of them were enterococci, with Enterococcus durans being the predominant species found. Pediococcus spp. were also isolated, together with strains of Streptococcus thermophilus. Within lactobacilli, 57% of the isolates were identified as Lactobacillus paracasei; Lact. curvatus, Lact. plantarum, Lact. fermentum, were present in a lower amount. The isolates were differentiated at strain-level by Rep-PCR analysis. This is the first effort to microbiological characterization of Valchiavenna's Bitto; the results suggest the possibility of preserving the wild bacterial population in order to protect the typical organoleptic characteristics of this traditional raw milk cheese and to select new strains for the dairy industry. PMID- 19810038 TI - Co-production of gamma-glutamylcysteine and glutathione by mutant strain Saccharomyces cerevisiae FC-3 and its kinetic analysis. AB - Co-production of gamma -glutamylcysteine (gamma -GC) and glutathione (GSH) by a novel mutant strain Saccharomyces cerevisiae FC-3 and its kinetic analysis were investigated. The strain could produce gamma -GC and GSH with high yields (4.22 and 14.3 mg/g-DCW, respectively) in batch submerged cultures. Effects of medium components and cultivation conditions on cell growth and the contents of intracellular gamma -GC and GSH were examined. Results show that 2% (w/v) sucrose and 2.5% (w/v) yeast extract are the best carbon and nitrogen sources, respectively, and supplement of three amino acids (glycine, cysteine and glutamate), each at 0.08% (w/v), in the medium could enhance gamma -GC and GSH production. In addition, optimal operating conditions are at the initial pH value of 7.0, 30 degrees C and 200 rev/min. Moreover, results obtained from kinetic analyses reveal that gamma -GC production is mixed-type growth associated, but GSH production is growth-associated. PMID- 19810039 TI - 2-dodecanol (decyl methyl carbinol) inhibits hyphal formation and SIR2 expression in C. albicans. AB - Candida albicans is capable of undergoing yeast-hypha transition to attain pathogenicity in humans. In this study, we investigated the differential expression of CaSIR2 via quantitative real-time PCR (qPCR), during yeast-hypha transition with and without the presence of 2-dodecanol. SIR2 transcript levels were found to be significantly enhanced after hyphal induction as compared to the yeast form. This study found that 2-dodecanol is able to inhibit hyphal development and block SIR2 up-regulation, even in hyphal-inducing growth conditions. We suggest that SIR2 may be involved in Candida albicans quorum sensing and serum-induced yeast-hyphae transition via the Ras1-cAMP-Efg1 signalling cascade. PMID- 19810040 TI - Isolation and identification of a Candida digboiensis strain from an extreme acid mine drainage of the Lignite Mine, Gujarat. AB - An extremely acidic mine drainage (AMD) water sample was collected in 1998 and 2008 from Panandhro lignite mine, Gujarat, India. The yeast isolated from this sample was identified using mini API identification system, as a member of genus Candida. The major cellular fatty acids detected by FAME from the isolate are C(16:0) and C(18:2) (cis 9,12)/C(18:0alpha) as 25.23 and 19.5%, respectively. The isolate was identified as Candida digboiensis by 18S rRNA gene sequence analysis and designated as Candida digboiensis SRDyeast1. Phylogenetic analysis using D1/D2 variable domains showed that the closest relative of this strain is Candida blankii with 3% divergence. This organism has been reported for the first time from the lignite mine AMD sample, and for cellular fatty acid analysis. This yeast is able to survive in the AMD sample preserved at 10-42 degrees C temperature since last 10 years along with iron oxidizing microorganisms. It can grow in the presence of 40% glucose, 10% NaCl and in the pH range of 1 to 10. The isolate is capable of producing enzymes like protease and lipase. This isolate differs from the type strain Candida digboiensis in as many as six physiological and metabolic characteristics. PMID- 19810041 TI - Multiresistant Vibrio cholerae non-O1/non-O139 from waters in South India: resistance patterns and virulence-associated gene profiles. AB - From different aquatic locations in Alleppey district, Kerala, South India a number (n = 36) of multiresistant non-O1, non-O139 V. cholerae strains were isolated. Water samples were filtered through 0.22 mum membrane filters, enriched in alkaline peptone water and plated onto thiosulfate-citrate-bile salts-sucrose (TCBS) agar. The isolates were resistance to cefotaxime (50%), nalidixic acid (44.4%), streptomycin and tetracycline (41.6%), trimethoprim (38.8%), co trimoxazole (33.3%), furazolidone (27.7%), neomycin and ofloxacin (19.4%), ciprofloxacin, norfloxacin and spectinomycin (16.6%), gentamicin (8.3%) and chloramphenicol (2.7%). To our knowledge, this is the first report from Kerala, South India on the emergence of multiple drug resistance in V. cholerae isolates belonging to serogroup other than O1 and O139. Virulence-associated gene profiling of the isolates by PCR revealed the presence of toxR (100%), rtxA (61.1%), hlyA (50%), mshA (33.3%), tcpA-acfB (13.8%) and st (2.7%) genes. The virulence gene clusters ctxA, ompU, ace, and zot were not detected. This study demonstrates the presence of a wide array of critical virulence factors in diverse strains of Vibrio cholerae non-O1/non-O139. Hence, this serogroup can no longer be ignored as an environmental reservoir of virulence genes. PMID- 19810042 TI - Characterization of new biosurfactant produced by Trichosporon montevideense CLOA 72 isolated from dairy industry effluents. AB - The yeast strain CLOA 72 isolated from the effluent of a dairy industry in Brazil and identified as Trichosporon montevideense, was able to grow and produce a glycolipid biosurfactant when cultured on a mineral medium (MM) with sunflower oil as the carbon source. Biosurfactant production was partially growth associated and maximal emulsification activity was observed at 144 h of cultivation (78.92%). The biosurfactant purified by precipitation with ethanol showed 78.66% emulsifying activity when used in concentrations above 4.5 mg/ml and was able to reduce the surface tension of water to values below 44.9 mN/m. The critical micellar concentration (CMC) was found to be 2.2 mg/ml. The highest emulsifying activity (E(24)) has been observed with vegetable oils, toluene, kerosene, isooctane, cyclohexane, hexane, diesel oil and hexadecane as compared to mineral oil and oleic acid. The biosurfactant also showed good stability during exposure to 100 degrees C for different periods of time (10 to 60 min), to high salinity (30% of NaCl, KCl and NaHCO(3)), and to a wide range of pH values (1-10). The biosurfactant purified by gel filtration chromatography is a glycolipid, with lipid portion containing 16.03% (9Z)-octadec-9-enoic acid, 14.92% hexadecanoic acid, and 9.63% (E) octadec-9-enoic acid and the carbohydrate portion containing mannose (35.29%), xylose (41.99%), arabinose (17.47%), and glucose (5.25%). PMID- 19810043 TI - Catalase and superoxide dismutase double staining zymogram technique for Deinococcus and Kocuria species exposed to multiple stresses. AB - Superoxide dismutase (SOD) and catalase expression is associated with oxidative stress. Existing techniques for the individual staining of SOD and catalase have been described in the past. The objective of this study was to achieve a simple and rapid technique for the double staining of bacterial SOD and catalase on the same polyacrylamide gel. SOD detection was carried out using nitro-blue tetrazolium (NBT) dye reduction followed by ferricyanide precipitation for negative staining of the catalase enzyme on the same gel. The staining procedure resulted in pale blue SOD bands while catalase appeared as yellow bands against a greenish blue background on the same gel. This technique was used to detect changes in the polymorphic forms of these enzymes in Deinococcus radiodurans R1 and Kocuria sp. C2 subjected to stresses like UV and gamma radiation and desiccation. PMID- 19810044 TI - Wild boars as reservoirs of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli of different phylogenetic groups. AB - ESBL-producing E. coli isolates have been isolated from eight of seventy seven faecal samples (10.4%) of wild boars in Portugal. The ESBL types identified by PCR and sequencing were bla(CTX-M-1) (6 isolates) and bla(CTX-M-1) + bla(TEM1-b) (2 isolates). Further resistance genes detected included tet (A) or tet (B) (in three tetracycline-resistant isolates), aad A (in three streptomycin-resistant isolates), cml A (in one chloramphenicol-resistant isolate), sul 1 and/or sul 2 and/or sul 3 (in all sulfonamide-resistant isolates). The intI 1 gene encoding class 1 integrase was detected in all ESBL-producing E. coli isolates. One isolate also carried the intI 2 gene, encoding class 2 integrase. The ESBL producing E. coli isolates could be assigned to phylogenetic groups B1 (3 isolates), B2 (3 isolates) or A (2 isolates). Amino acid change in GyrA protein (Ser83Leu or Asp87Tyr) was detected in three nalidixic acid-resistant and ciprofloxacin-susceptible isolates. Two amino acid changes in GyrA (Ser83Leu + Asp87Asn) and one in ParC (Ser80Ile) were identified in two nalidixic acid- and ciprofloxacin-resistant isolates. As evidenced by this study wild boars could be a reservoir of antimicrobial resistance genes. PMID- 19810045 TI - Spontaneous and protein-induced secretion of proteinases from Saccharomyces cerevisiae. AB - Many fungi are capable of secreting the wide spectrum of hydrolytic enzymes. We characterized an inducible proteinase secretion in yeasts, Saccharomyces cerevisiae. The proteinase secretion by S. cerevisiae was induced in the presence of yeast extract, or of purified proteins, such as bovine serum albumin, casein, or ovalbumin, and some proteolytic activity was present also without protein inducer. We found that properties of proteinases induced under cultivation conditions were different in various aspects (temperature- and pH-dependencies, substrate specificities, sensitivities to proteinase inhibitors). Proteinase activities were also characterized by gelatin zymography. Multiple proteinase bands with wide-molecular weights (ranging from 45 to 240 kDa) were detected and patterns of proteinase bands were different. S. cerevisiae cells were able to retain the information about previous contacts with protein inducer resulting in faster and more intensive proteinase secretion response after repeated induction. PMID- 19810046 TI - Rapid and sensitive detection of Mollicutes in cell culture by polymerase chain reaction. AB - Infections with Mollicutes species (such as Mycoplasma, Acholeplasma, and Ureaplasma) can induce a variety of problems in living organisms and laboratory cell cultures. Therefore, it is necessary to establish a routine diagnostic protocol for Mycoplasma infection in order to ensure reliable research results, as well as the safety of commercial biological products. For that purpose a novel PCR-based procedure using specific designed primers complementary to 16S rRNA genome region of mollicute species was evaluated. PCR was optimized and sensitivity and specificity was evaluated by defined cell count concentrations (2 31250 CFU/ml) of different strains of Mycoplasma, Acholeplasma and Ureaplasma. Amplicon (272 bp) was cloned by PCR-cloning and sequenced by dideoxy chain termination. PCR, was found to be able to detect 10 copies of mollicute target DNA. No cross-reactivity with genomic DNA of non-mollicute bacteria or human cell lines was observed. Forty seven human and animal cell lines were evaluated for mollicute contamination. Twenty five cell lines (53%) were correctly identified as contaminated by this molecular approach. The results of this study demonstrated that this PCR-based method is not only fast and reproducible, but also highly sensitive and specific for detecting contaminant mycoplasmas in cell cultures. PMID- 19810047 TI - Detection and characterization of class 1 integrons in Aeromonas spp. isolated from human diarrheic stool in Mexico. AB - We determined the presence of class 1 integrons related to the acquisition of resistance to antimicrobials in Aeromonas spp. isolated from individuals with diarrhea. Species were identified as A. caviae, A. hydrophila, A. veronii and A. media using PCR-RFLP of the 16S rDNA. Selected isolates were further characterized by ERIC-PCR. Resistance to chloramphenicol, aztreonam, tetracycline, trimethoprim/sulfamethoxazole, nalidixic acid and streptomycin, among others, was determined using the Kirby-Bauer method. Integrons were detected by PCR amplification of the 5' conserved, variable, and 3' conserved regions. Sequencing of the variable regions revealed class 1 integrons with cassettes encoding resistance to trimethoprim (dfrA12, dfrA15, dfrB4), streptomycin/spectinomycin (aadA2, aadA1), oxacillin (oxa2) and chloramphenicol (catB3, cmlA4). Others had an open reading frame (orfD) or no insert at all. To our knowledge, this is the first description of the occurrence of genes cmlA4 and dfrA15 in Aeromonas class 1 integrons. Not all the integron-linked cassettes conferred their associated resistances, which suggests the inactivity of some cassettes. Most integrons were chromosomally located. The presence of class 1 integrons similar to those found in a wide variety of bacterial genera from different origins, including environmental and fish-borne Aeromonas, confirms the stability and horizontal transfer of these genetic elements. PMID- 19810048 TI - Detection of antibiotic resistant E. coli and Enterococcus spp. in stool of healthy growing children in Portugal. AB - From stool specimens of 118 healthy children's (1-14 years) in Portugal 92 E. coli and 101 Enterococcu s spp. strains have been isolated. Almost half (40.2%) of the E. coli isolates were resistant to ampicillin, 25.0% were resistant to tetracycline and 26.1% were resistant to streptomycin. Resistance genes detected by specific PCR included bla(TEM) and/or bla(SHV) and/or bla(CTX-M) (33 of 37 ampicillin and/or cefotaxime resistant isolates), tet (A) and/or tet (B) (16 of 23 tetracycline-resistant isolates), aad A (19 of 24 streptomycin-resistant isolates), cml A (in the two chloramphenicol-resistant isolates), aac (3)-II with/without aac (3)-IV (in the four gentamicin-resistant isolates), sul 1 and/or sul 2 and/or sul 3 (in all trimethoprim/sulfamethoxazole resistant isolates). The majority of the resistant E. coli isolates (69.1%) belonged to phylogenetic group B2. Of the enterococci isolates E. faecium (n = 53), E. faecalis (n = 41), E. hirae (n = 4) and E. durans (n = 3) more than one-fourth (28.7%) of the isolates were resistant to tetracycline; 21.8% were resistant to erythromycin and 8.9% were resistant to kanamycin. Resistance genes detected by PCR in enterococci included aph (3)'-IIIa (in all kanamycin-resistant isolates), aac (6') (in all gentamicin-resistant isolates), tet (M) and/or tet (L) (26 of 29 tetracycline resistant isolates), erm (B) (17 of 22 erythromycin-resistant isolates). This survey showed that faecal bacteria such as E. coli and enterococci of healthy growing children's could be a reservoir of antimicrobial resistance genes. PMID- 19810049 TI - Elevated curdlan production by a mutant of Agrobacterium sp. ATCC 31749. AB - A mutant strain of the curdlan-producing bacterium Agrobacterium sp. ATCC 31749, isolated by ethylmethane sulfonate mutagenesis and resistance to ampicillin, was capable of elevated curdlan synthesis. Using 2.5% corn syrup, glucose or maltose as a carbon source, the mutant strain was shown to produce a 1.5-fold, 1.5-fold or 1.5-fold higher level of curdlan, respectively, than its parent strain after 120 h of growth. The mutant strain produced higher curdlan levels after 96 or 120 h of growth on glucose or maltose as a carbon source than it did on corn syrup. Biomass production by the mutant strain grown on the carbon sources studied was slightly elevated compared to its parent strain. It was concluded that the elevated curdlan production observed for the mutant strain grown on corn syrup or glucose was not due to an increase in biomass production. PMID- 19810050 TI - Interference of chromium with biological systems in yeasts and fungi: a review. AB - This paper deals with the interactions of chromium (Cr) with biological systems, focusing in particular on yeasts and fungi. These interactions are analysed with primarily regard to biochemical functions, but higher levels of organization are also considered. Thus, the morphological and cytological characteristics of selected microorganisms in response to exposure to chromium ions are evaluated. The different oxidation states of chromium and reactive oxygen species (ROS) generated in redox reactions with chromium ions are presented and characterized. The interactions of the most exposed subcellular structures, including the cell wall, plasma membrane and nuclei, have been deeply investigated in recent years, for two major reasons. The first is the toxicity of chromium ions and their strong impact on the metabolism of many species, ranging from microbes to humans. The second is the still disputed usefulness of chromium ions, and in particular trivalent chromium, in the glucose and fat metabolisms. Chromium pollution is still an important issue in many regions of the world, and various solutions have been proposed for the bioremediation of soil and water with selected microbial species. Yeasts and especially moulds have been most widely investigated from this aspect, and the biosorption and bioaccumulation of chromium for bioremediation purposes have been demonstrated. Accordingly, the mechanisms of chromium tolerance or resistance of selected microbes are of particular importance in both bioremediation and waste water treatment technologies. The mechanisms of chromium toxicity and detoxification have been studied extensively in yeasts and fungi, and some promising results have emerged in this area. PMID- 19810051 TI - Domain-ligand mapping for enzymes. AB - In this paper we provide an overview of our current knowledge of the mapping between small molecule ligands and protein domains. We give an overview of the present data resources available on the Web, which provide information about protein-ligand interactions, as well as discussing our own PROCOGNATE database. We present an update of ligand binding in large protein superfamilies and identify those ligands most frequently utilized by nature. Finally we discuss potential uses for this type of data. PMID- 19810052 TI - Determination of metabolite profiles in tropical wines by 1H NMR spectroscopy and chemometrics. AB - Traditionally, wines are produced in temperate climate zones, with one harvest per year. Tropical wines are a new concept of vitiviniculture that is being developed, principally in Brazil. The new Brazilian frontier is located in the northeast region (Sao Francisco River Valley) in Pernambuco State, close to the equator, between 8 and 9 degrees S. Compared with other Brazilian and worldwide vineyards, the grapes of this region possess peculiar characteristics. The aim of this work is a preliminary study of commercial Sao Francisco River Valley wines, analyzing their metabolite profiles by (1)H NMR and chemometric methods. PMID- 19810053 TI - Assessing cytotoxicity of (iron oxide-based) nanoparticles: an overview of different methods exemplified with cationic magnetoliposomes. AB - Iron oxide nanoparticles are the most widely used T(2)/T(2)* contrast agents and for biomedical research purposes, one of the main applications is the in vitro labeling of stem or therapeutic cells, allowing them to be subsequently tracked in vivo upon transplantation. To allow this, the nanoparticles used should not show any sign of cytotoxicity and not affect cellular physiology as this could impede normal cell functionality in vivo or lead to undesired side-effects. Assessing the biocompatibility of the nanoparticles has proven to be quite a difficult task. In the present work, a small overview of commonly used assays is presented in order to assess several aspects, such as cell viability, induction of reactive oxygen species, nanoparticle uptake, cellular morphology, cellular proliferation, actin cytoskeleton architecture and differentiation of stem cells. The main focus is on comparing the advantages and disadvantages of the different assays, highlighting several common problems and presenting possible solutions to these problems as well as pointing out the high importance of the relationship between intracellular nanoparticle concentration and cytotoxicity. PMID- 19810054 TI - PCR experion automated electrophoresis system to detect Listeria monocytogenes in foods. AB - Listeria monocytogenes is frequently found as a contaminant in raw and ready-to eat foods. The ability of L. monocytogenes to multiply at refrigeration temperatures and to grow in a wide range of pH values is of particular concern for food safety. According to the European Union regulation on microbiological criteria for foodstuffs, L. monocytogenes must be absent in some categories of ready-to-eat foods. The standard microbiological method for L. monocytogenes detection in foods (ISO 11290-1: 1996 (ISO, International Organization for Standardization)) is cost and time consuming. Developments of rapid, cost effective and automated diagnostic methods to detect food-borne pathogens in foods continue to be a major concern for the industry and public health. The aim of this study was the development of a rapid, sensitive and specific molecular detection method for L. monocytogenes. To this purpose, we have applied a capillary electrophoresis method to a PCR protocol (PCR-EES (EES, experion automated electrophoresis system)) for detecting L. monocytogenes in food. In particular, a microfluidic chip-based automated electrophoresis system (experion automated electrophoresis system, Bio-Rad Laboratories, USA) was used for the rapid and automatic analysis of the amplicons. Fifty naturally contaminated samples were analysed with this method and the results were compared with those obtained with ISO method. Moreover, the microfluidic chip-based automated electrophoresis system was compared with classical gel electrophoresis (PCR-CGE). The results showed that after 24 h of culture enrichment, the PCR-EES showed a relative accuracy of 100% with ISO, while using PCR-CGE decreased it down to 96%. After 48 h of enrichment, both PCR-EES and PCR-CGE showed an accuracy of 100% with ISO. PMID- 19810055 TI - Application of LC-NMR to analysis of carotenoids in foods. AB - LC-NMR has been applied to componential analysis of carotenoids in several foods, specifically, tomato juice, palm oil, and satsuma mandarin orange juice. The crude carotenoids extracted with organic solvent from these foodstuffs were analyzed after simple pre-processing. Three, four, and two kinds of carotenoids were identified for tomato juice, palm oil, and satsuma mandarin orange, respectively, primarily by comparing their NMR spectra with those of pure standard. PMID- 19810056 TI - Hydrogen-atom abstraction reactions by manganese(V)- and manganese(IV)-oxo porphyrin complexes in aqueous solution. AB - High-valent manganese(IV or V)-oxo porphyrins are considered as reactive intermediates in the oxidation of organic substrates by manganese porphyrin catalysts. We have generated Mn(V)- and Mn(IV)-oxo porphyrins in basic aqueous solution and investigated their reactivities in C-H bond activation of hydrocarbons. We now report that Mn(V)- and Mn(IV)-oxo porphyrins are capable of activating C-H bonds of alkylaromatics, with the reactivity order of Mn(V) oxo>Mn(IV)-oxo; the reactivity of a Mn(V)-oxo complex is 150 times greater than that of a Mn(IV)-oxo complex in the oxidation of xanthene. The C-H bond activation of alkylaromatics by the Mn(V)- and Mn(IV)-oxo porphyrins is proposed to occur through a hydrogen-atom abstraction, based on the observations of a good linear correlation between the reaction rates and the C-H bond dissociation energy (BDE) of substrates and high kinetic isotope effect (KIE) values in the oxidation of xanthene and dihydroanthracene (DHA). We have demonstrated that the disproportionation of Mn(IV)-oxo porphyrins to Mn(V)-oxo and Mn(III) porphyrins is not a feasible pathway in basic aqueous solution and that Mn(IV)-oxo porphyrins are able to abstract hydrogen atoms from alkylaromatics. The C-H bond activation of alkylaromatics by Mn(V)- and Mn(IV)-oxo species proceeds through a one-electron process, in which a Mn(IV)-oxo porphyrin is formed as a product in the C-H bond activation by a Mn(V)-oxo porphyrin, followed by a further reaction of the Mn(IV)-oxo porphyrin with substrates that results in the formation of a Mn(III) porphyrin complex. This result is in contrast to the oxidation of sulfides by the Mn(V)-oxo porphyrin, in which the oxidation of thioanisole by the Mn(V)-oxo complex produces the starting Mn(III) porphyrin and thioanisole oxide. This result indicates that the oxidation of sulfides by the Mn(V)-oxo species occurs by means of a two-electron oxidation process. In contrast, a Mn(IV)-oxo porphyrin complex is not capable of oxidizing sulfides due to a low oxidizing power in basic aqueous solution. PMID- 19810057 TI - Self-assembling of Er(2)O(3)-TiO(2) mixed oxide nanoplatelets by a template-free solvothermal route. AB - An easy solvothermal route has been developed to synthesize the first mesoporous Er(2)O(3)-TiO(2) mixed oxide spherical particles composed of crystalline nanoplatelets, with high surface area and narrow pore size distribution. This synthetic strategy allows the preparation of materials at low temperature with interesting textural properties without the use of surfactants, as well as the control of particle size and shape. TEM and Raman analysis confirm the formation of nanocrystalline Er(2)O(3)-TiO(2) mixed oxide. Mesoscopic ordered porosity is reached through the thermal decomposition of organic moieties during the synthetic process, thus leading to a template-free methodology that can be extended to other nanostructured materials. High specific surface areas (up to 313 m(2) g(-1)) and narrow pore size distributions are achieved in comparison to the micrometric material synthesized by the traditional sol-gel route. This study opens new perspectives in the development, by solvothermal methodologies, of multifunctional materials for advanced applications by improving the classical pyrochlore properties (magnetization, heat capacity, catalysis, conductivity, etc.). In particular, since catalytic reactions take place on the surface of catalysts, the high surface area of these materials makes them promising candidates for catalysts. Furthermore, their spherical morphology makes them appropriate for advanced technologies in, for instance, ceramic inkjet printers. PMID- 19810058 TI - Inverted opal fluorescent film chemosensor for the detection of explosive nitroaromatic vapors through fluorescence resonance energy transfer. AB - This paper reports an inverted opal fluorescence chemosensor for the ultrasensitive detection of explosive nitroaromatic vapors through resonance energy-transfer-amplified fluorescence quenching. The inverted opal silica film with amino ligands was first fabricated by the acid-base interaction between 3 aminopropyltriethoxysilane and surface sulfonic groups on polystyrene microsphere templates. The fluorescent dye was then chemically anchored onto the interconnected porous surface to form a hybrid monolayer of amino ligands and dye molecules. The amino ligands can efficiently capture vapor molecules of nitroaromatics such as 2,4,6-trinitrotoluene (TNT) through the charge-transfer complexing interaction between electron-rich amino ligands and electron-deficient aromatic rings. Meanwhile, the resultant TNT-amine complexes can strongly suppress the fluorescence emission of the chosen dye by the fluorescent resonance energy transfer (FRET) from the dye donor to the irradiative TNT-amino acceptor through intermolecular polar-polar resonance at spatial proximity. The quenching response of the highly ordered porous films with TNT is greatly amplified by at least 10-fold that of the amorphous silica films, due to the interconnected porous structure and large surface-to-volume ratio. The inverted opal film with a stable fluorescence brightness and strong analyte affinity has lead to an ultrasensitive detection of several ppb of TNT vapor in air. PMID- 19810059 TI - Mechanistic studies of C-C bond cleavage of nitriles by dinuclear metal cryptates. AB - We previously reported that dinuclear copper(II) cryptate [Cu(2)L](4+) cleaves the C-C bond of acetonitrile at room temperature to produce a cyano-bridged dinuclear cryptate and methanol, whereby the reaction mechanism has not yet become clear. We have now systemically investigated this reaction, and four cryptates, [Cu(2)L](ClO(4))(4) (1), [Zn(2)L](ClO(4))(4) (2), [Cu(2)L(H(2)O)(2)](CF(3)SO(3))(4) (5), and [Cu(2)L(OH)(OH(2))](ClO(4))(3) (6) are reported here. Cryptates 1 and 2 can cleave the C--C bonds of acetonitrile, propionitrile, and benzonitrile at room temperature under open atmospheric conditions to give cyano-bridged cryptates [Cu(2)L(CN)](ClO(4))(3) (3) and [Zn(2)L(CN)](ClO(4))(3) (4), respectively, and the corresponding alcohol. In contrast, 5 and 6 do not show any C-C bond activation of nitriles, as the interior axial positions of Cu(II) in 5 and 6 are occupied by water/OH(-). The C C bond cleavage of (S)-(+)-2-methylbutyronitrile by 2 produced (R)-(-)-2-butanol only; that is, the cleavage reaction proceeds through an S(N)2 pathway (Walden inversion). PMID- 19810060 TI - Ionic liquids made with dimethyl carbonate: solvents as well as boosted basic catalysts for the michael reaction. AB - This article describes 1) a methodology for the green synthesis of a class of methylammonium and methylphosphonium ionic liquids (ILs), 2) how to tune their acid-base properties by anion exchange, 3) complete neat-phase NMR spectroscopic characterisation of these materials and 4) their application as active organocatalysts for base-promoted carbon-carbon bond-forming reactions. Methylation of tertiary amines or phosphines with dimethyl carbonate leads to the formation of the halogen-free methyl-onium methyl carbonate salts, and these can be easily anion-exchanged to yield a range of derivatives with different melting points, solubility, acid-base properties, stability and viscosity. Treatment with water, in particular, yields bicarbonate-exchanged liquid onium salts. These proved strongly basic, enough to efficiently catalyse the Michael reaction; experiments suggest that in these systems the bicarbonate basicity is boosted by two orders of magnitude with respect to inorganic bicarbonate salts. These basic ionic liquids used in catalytic amounts are better even than traditional strong organic bases. The present work also introduces neat NMR spectroscopy of the ionic liquids as a probe for solute-solvent interactions as well as a tool for characterisation. Our studies show that high catalytic efficacy of functional ionic liquids can be achieved by integrating their green synthesis, along with a fine-tuning of their structure. Demonstrating that ionic liquid solvents can be made by a truly green procedure, and that their properties and reactivity can be tailored to the point of bridging the gap between their use as solvents and as catalysts. PMID- 19810061 TI - Lanthanide-alkali metal sandwich complexes: synthesis, structure, and solvent mediated redox transformations, and one-dimensional frameworks assembled through cation-arene pi interactions. AB - Reaction of the potassium salt of amido ligand [Ph(2)Si(NAr)(2)](2-) (L, Ar = 2,6 iPr(2)C(6)H(3)) and LnI(2)(thf)(2) (Ln = Sm, Yb) gives sandwich complexes [(L)(2)Ln{K(Et(2)O)}(2)] (Ln = Sm (2), Yb (3)) with potassium-arene pi interactions. Reaction of 2 with azobenzene gave the dimeric samarium cluster [(L)(2)Sm(2)(mu-eta(2):eta(2)-N(2)Ph(2))(2){K(thf)(2)}(2)] (4) and the tetrameric [(L)Sm(4) (mu-eta(2):eta(2)-N(2)Ph(2))(3) (mu(3)-NPh)(2)(thf)(3)] (5). On the other hand, the reaction of 2 with alpha-diimines ligands ArN=CRCR=NAr (DAD, R = H, Me) gives two Sm(III) complexes: polymeric [(L)Sm{(ArN)RC=CR(NAr)}K](n) (R = H (6), Me (7)) assembled through cation-pi interactions and byproduct [{(L)(2)Sm}{K(thf)(6)}] (8). Complexes 2-8 have been fully characterized by elemental analyses and X-ray crystallography. In particular, crystallographic analyses of 6 and 7 revealed that in both complexes samarium(III) is stabilized by dianionic DAD units. PMID- 19810062 TI - Isotope-sensitive degenerate [1,3]-hydrogen migration versus competitive enol keto tautomerization. PMID- 19810063 TI - Methionine can favor DNA platination by trans-coordinated platinum antitumor drugs. PMID- 19810064 TI - Nanostructuring of patterned microelectrodes to enhance the sensitivity of electrochemical nucleic acids detection. PMID- 19810065 TI - Direct, electronic microRNA detection for the rapid determination of differential expression profiles. PMID- 19810066 TI - The continuous-flow synthesis of Ibuprofen. PMID- 19810067 TI - Trifluoromethanesulfanylamides as easy-to-handle equivalents of the trifluoromethanesulfanyl cation (CF(3)S(+)): reaction with alkenes and alkynes. PMID- 19810068 TI - Two-color reversible switching in a photochromic ruthenium sulfoxide complex. PMID- 19810069 TI - Silicon compounds of neon and argon. PMID- 19810070 TI - Polyelectrolyte microcapsules as antigen delivery vehicles to dendritic cells: uptake, processing, and cross-presentation of encapsulated antigens. PMID- 19810071 TI - Cross-linking of transmembrane helices reveals a rigid-body mechanism in bacteriorhodopsin transport. PMID- 19810072 TI - Siamese depsipeptides: constrained bicyclic architectures. PMID- 19810073 TI - Chemistry with bare silicon clusters in solution: a transition-metal complex of a polysilicide anion. PMID- 19810075 TI - Organic chemistry: J. M. J. Frechet awarded / catalysis: K. Nozaki honored / organometallic chemistry: Prize for P. Braunstein. PMID- 19810074 TI - Metal-organic hendecahedra assembled from dinuclear paddlewheel nodes and mixtures of ditopic linkers with 120 and 90 degrees bend angles. PMID- 19810078 TI - C5-functionalized LNA: unparalleled hybridization properties and enzymatic stability. PMID- 19810079 TI - Discovery of a small-molecule inhibitor of the KIX-KID interaction. PMID- 19810080 TI - FT Raman and DFT study on a series of all-anti oligothienoacenes end-capped with triisopropylsilyl groups. AB - Herein, we study the pi-conjugational properties of a homologous series of all anti oligothienoacenes containing four to eight fused thiophene rings by means of FT Raman spectroscopy and DFT calculations. The theoretical analysis of the spectroscopic data provides evidence that selective enhancement of a very limited number of Raman scatterings is related to the occurrence in these oligothienoacenes of strong vibronic coupling between collective nu(C=C) stretching modes in the 1600-1300 cm(-1) region and the HOMO/LUMO frontier orbitals (HOMO=highest occupied molecular orbital; LUMO=lowest unoccupied molecular orbital). The correlation of the Raman spectroscopic data and theoretical results for these all-anti oligothienoacenes with those previously collected for a number of all-syn oligothienohelicenes gives further support to the expectation that cross-conjugation is dominant in heterohelicenes. Fully planar all-anti oligothienoacenes display linear pi conjugation which seemingly does not reach saturation with increasing number of annulated thiophene rings in the oligomeric chain at least up to the octamer. PMID- 19810081 TI - Influence of confined water on the photophysics of dissolved solutes in reverse micelles. AB - The photophysical parameters of two probes with largely different hydrophobic character, namely, coumarin 1 and coumarin 343, are investigated in sodium bis-(2 ethylhexyl)sulfosuccinate (AOT)/hexane/water reverse micelles at various water/AOT molar ratio w(0). Correlation of photophysical parameters such as fluorescence quantum yield, fluorescence lifetime, and emission maxima with w(0) indicate distinctly different trends below and above w(0) approximately 7 for both probes. The variation of the average rotational correlation times obtained from fluorescence anisotropy decays for both probes in reverse micelles further corroborate the above observation. Similar studies were also performed in nonaqueous reverse micelles with acetonitrile as polar solvent. Similar to aqueous reverse micelles, breaks in the photophysical parameters with increasing acetonitrile/AOT molar ratios w'(0) were also observed in these cases, although at a much lower w'(0) value of 3. The present results indicate that around w(0) approximately 7 for aqueous reverse micelles (and around w'(0) approximately 3 for nonaqueous reverse micelles) a distinct change occurs in the probe microenvironment, which is rationalized on the basis of the relative populations of interfacial and core water. We propose that until the ionic head groups and counterions are fully solvated by polar solvents, that is, up to w(0) approximately 7 (or w'(0) approximately 3), the interfacial water population dominates. Above these molar ratios coalescence of excess water molecules with each other to form truncated H-bonded water clusters leads to a sizable population of core water. This is further substantiated by changes in the IR absorption spectra for the O--D stretching mode of diluted D(2)O in reverse micelles with varying w(0). Critical comparison of the present results with relevant literature reports provide clear support for the proposals made on water structure in reverse micelles. The role of relative size of the probe and the reverse micelles for differences in polar solvent to AOT ratios (w(0)=7 and w'(0)=3) in the observed breaks in the two types of reverse micelles is also discussed. PMID- 19810082 TI - A new class of boron nanotube. AB - The configurations, stability and electronic structures of a new class of boron sheet and related boron nanotubes are predicted within the framework of density functional theory. This boron sheet is sparser than those of recent proposals. Our theoretic results show that the stable boron sheet remains flat and is metallic. There are bands similar to the pi-bands in graphite near the Fermi level. Stable nanotubes with various diameters and chiral vectors can be rolled from the sheet. Within our study, only the thin (8, 0) nanotube with a band gap of 0.44 eV is semiconducting, while all the other thicker boron nanotubes are metallic, independent of their chirality. It indicates the possibility, in the design of nanodevices, to control the electronic transport properties of the boron nanotube through the diameter. PMID- 19810083 TI - Selective guest docking in metal-organic framework materials. PMID- 19810084 TI - Modelling atypical small-molecule mimics of an important stem cell cytokine, thrombopoietin. AB - We report the first comprehensive 3D QSAR study of a large, structurally diverse set of compounds that act as atypical thrombopoietin (TPO) mimics by interacting with the transmembrane domain of the TPO receptor, c-MPL. These agonists of c-MPL were superimposed according to a pharmacophore hypothesis, resulting in 3D QSAR models of high statistical significance. The pharmacophore-based superimposition and comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were used to derive the QSAR models relating structure to the published in vitro bioactivities of the TPO mimics. The CoMFA and CoMSIA models gave high correlation coefficients of the bioactivities with the derived fields, resulting in robust prediction of agonist activity of the superimposed compounds. The models have been interpreted in terms of the requirements for binding to the transmembrane domain of the TPO receptor. PMID- 19810085 TI - Tin oxide thin film with three-dimensional ordered reticular morphology as a lithium ion battery anode. AB - Three-dimensional (3D) reticular SnO(2) thin films deposited on copper and stainless steel substrates were prepared by the electrostatic spray deposition (ESD) technique. The 3D reticular SnO(2) film exhibit a high reversible capacity near 300 mAh g(-1) up to the 50th cycle. PMID- 19810087 TI - Thrombopoietin and thrombocytopenia in anorexia nervosa with severe liver dysfunction. AB - OBJECTIVE/METHOD: Patients with anorexia nervosa (AN) display various physical complications. One such problem is thrombocytopenia. However, no studies have investigated links between AN and thrombopoietin (TPO), which is the primary regulator of megakaryocyte and platelet production produced mainly in the liver, although TPO has been recently reported to play a role in thrombocytopenia in liver diseases. We therefore clarified temporal relationships between platelet counts and TPO level in a woman with AN and severe liver dysfunction in whom platelet count decreased to 53 x 109/L. RESULTS: While international normalized ratio (INR) was low, serum TPO level was relatively low despite low platelet counts. After INR and liver enzymes improved in conjunction with improved nutritional status, a rapid increase in TPO was observed and platelet count began to rise. DISCUSSION: These findings thus suggest that decreased TPO production accompanying liver dysfunction may be related to thrombocytopenia besides myelosuppression in AN with malnutrition. PMID- 19810088 TI - Identifying PTSD personality subtypes in a workplace trauma sample. AB - The authors sought to identify personality clusters derived from the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) Personality Psychopathology Five Scales in a sample of workplace claimants with posttraumatic stress disorder (PTSD). Three clusters--low pathology, internalizing, and externalizing were recovered similar to those obtained by M. W. Miller and colleagues (2003, 2004, 2007) in samples of combat veterans and sexual assault victims. Internalizers and externalizers scored comparably on measures of PTSD symptom severity, general distress, and negative affect. Internalizers were uniquely characterized by anhedonia and depressed mood; externalizers by antisocial behavior, substance abuse, and anger/aggression. PMID- 19810089 TI - CTSA program continues 'on track' despite backlog. PMID- 19810091 TI - Balancing risk and reward: the question of natalizumab. PMID- 19810092 TI - Comments on 'choosing an optimal method to combine P-values': by S. Won, N. Morris, Q. Liu, R. C. Elston, Statistics in Medicine 2009; 28:1537-1553. PMID- 19810093 TI - Sphingosine-1-phosphate receptor type 1 regulates glioma cell proliferation and correlates with patient survival. AB - Sphingosine-1-phosphate (S1P) is a bioactive lipid that signals through a family of G protein-coupled receptors consisting of 5 members termed S1P(1-5), and it regulates cellular proliferation, migration and survival. We investigated the expression and role of S1P receptors in glioma. Human glioma expressed S1P(1), S1P(2), S1P(3), and S1P(5) by quantitative real-time PCR analysis. Expression of the S1P(1) was significantly lower in glioblastoma than in the normal brain (p < 0.01) and diffuse astrocytoma (p < 0.05). Immunoblotting showed that normal brain expressed more S1P(1) protein than did glioblastoma. Immunohistochemistry showed that S1P(1) was localized predominantly in the astrocytes in the normal brain, but no staining was observed in glioblastoma. Downregulation of S1P(1) expression correlated with poor survival of patients with glioblastoma (p < 0.05). S1P(1) small interfering RNA promoted cell proliferation in high-expressor glioma cell lines (T98G, G112). Cell proliferation was promoted by the pertussis toxin, which deactivates G(i/o) type of G proteins; the S1P(1) is exclusively coupled to these proteins. Forced expression of the S1P(1) in low-expressor cell lines (U87, U251) resulted in decreased cell growth and led to suppressed tumor growth in transplanted gliomas in vivo. Furthermore, we found a significant association between the S1P(1) expression and early growth response-1, a transcriptional factor that exhibits tumor suppression in glioblastoma cells (p < 0.05). These data indicate that the downregulation of S1P(1) expression enhances the malignancy of glioblastoma by increasing cell proliferation and correlates with the shorter survival of patients with glioblastoma. PMID- 19810094 TI - A specific cytotoxic T-lymphocyte epitope presentation system for antitumor immunity. AB - The magnitude of CTL-mediated immunity response is highly dependent on the density of antigenic peptide-MHC I complexes at the cell surface. In this study, we adopt a novel strategy to promote the surface level of specific peptide-MHC I complexes. The strategy combines the inhibition of transporter associated with antigen processing (TAP) with the delivery of specific peptide into endoplasmic reticulum directly without the help of TAP. First, RNA interference (RNAi) technology was used to inhibit TAP expression for blocking endogenous epitope assembled MHC class I on cell surface. Second, a peptide epitope of interest was covalently linked onto human beta-2-microglobulin (beta2m). Both TAP-specific siRNA and the peptide-linked beta2m were delivered into antigen-presentation cells sequentially or simultaneously using a retrovirus delivery system. The combined strategy produces a significant amount of MHC I loaded with specific epitopes on the surface while reducing endogenously peptide-assembled MHC class I both in vitro and in vivo. The efficacy of induction of specific immune response with the strategy against tumor cells is demonstrated in both tumor cell lines and a syngenic graft tumor model. PMID- 19810095 TI - Salivary gland and nasopharyngeal cancers in individuals with acquired immunodeficiency syndrome in United States. AB - Individuals with acquired immunodeficiency syndrome (AIDS) manifest an increased risk of cancer, particularly cancers caused by oncogenic viruses. Because some salivary gland and nasopharyngeal cancers are associated with Epstein Barr virus, the impact of AIDS on these cancers needs further evaluation. We used linked U.S. AIDS and cancer registry data (N = 519,934 people with AIDS) to derive standardized incidence ratios (SIRs) comparing risk of salivary gland and nasopharyngeal cancers to the general population. For salivary gland cancers (N = 43 cases), individuals with AIDS had strongly elevated risks for lymphoepithelial carcinoma (SIR 39, 95% CI 16-81) and squamous cell carcinoma (SIR 4.9, 95% CI 2.5 8.6). Among nasopharyngeal cancers (N = 39 cases), risks were elevated for both keratinizing and nonkeratinizing carcinomas (SIR 2.4, 95% CI 1.5-3.7 and SIR 2.4, 95% CI 1.2-4.4, respectively). The elevated risks of salivary gland and nasopharyngeal cancers among people with AIDS suggest that immunosuppression and oncogenic viral infections are etiologically important. PMID- 19810096 TI - Combination of p53 codon 72 polymorphism and inactive p53 mutation predicts chemosensitivity to 5-fluorouracil in colorectal cancer. AB - There are increasing reports showing the clinical significance of the p53 polymorphism status in terms of the response to chemotherapy. We investigated whether p53 polymorphism and mutation were associated with in vitro sensitivity to 5-fluorouracil (5-FU) in patients with colorectal cancer. Chemosensitivity to 5-FU was evaluated by the collagen gel droplet embedded culture drug sensitivity test. 5-FU sensitivity of tumor cells without inactive p53 mutation in the arginine/arginine (Arg/Arg) variant was significantly higher than that of tumor cells with or without inactive p53 mutation in other variants (p = 0.022), whereas the 5-FU sensitivity of tumor cells with inactive p53 mutation in the Arg/Arg variant was significantly lower than that of tumor cells with or without inactive p53 mutation in other variants (p = 0.002). In the Arg/Arg variant, apoptotic cells induced by 5-FU treatment in patients without inactive p53 mutation were more markedly increased than those in patients with inactive p53 mutation (p = 0.037). Bax and Bcl-2 protein expressions in tumor tissue treated with 5-FU were associated with both 5-FU sensitivity and the apoptotic cell count. Our data show that the Arg/Arg genotype without inactive p53 mutation could be predictive of a more favorable response and the Arg/Arg genotype with inactive p53 mutation a less favorable response to chemotherapy using 5-FU in CRC. The combination of the p53 codon 72 polymorphism and p53 mutation status is a potential predictive marker of sensitivity to 5-FU in CRC. PMID- 19810097 TI - Preferential recruitment of interferon-gamma-expressing TH17 cells in multiple sclerosis. AB - OBJECTIVE: There is substantial evidence supporting the role of interferon (IFN) gamma-producing T helper (T(H)) 1 and interleukin (IL)-17-expressing T(H)17 lymphocytes in multiple sclerosis (MS) and its animal model, experimental allergic encephalomyelitis (EAE). However, to date little is known about the potential cooperative interplay between these 2 cytokines. In the current study, we sought to evaluate the frequency of IFN-gamma-expressing T(H)17 lymphocytes in MS and EAE, and study their recruitment into the central nervous system (CNS). METHODS: Human T(H)17 lymphocytes were expanded in vitro from the blood of healthy controls and relapsing MS patients using IL-23. Immune cell migration to the CNS was assessed in vitro with primary cultures of human blood-brain barrier (BBB)-derived endothelial cells, and in vivo in EAE mice. RESULTS: We demonstrate that in response to IL-23, human memory lymphocytes expand into a T(H)17 phenotype, with a subpopulation of cells simultaneously expressing IFN-gamma and IL-17. We note that lymphocytes obtained from the blood of relapsing MS patients have an increased propensity to expand into IFN-gamma-producing T(H)17 cells and identify numerous T lymphocytes coexpressing IL-17 and IFN-gamma in brain tissue of MS patients. We also find lymphocytes expressing both the T(H)1- and the T(H)17-associated transcription factors ROR gamma t and T-bet, in situ and in vitro. We further provide in vitro and in vivo evidence that IFN-gamma(+) T(H)17 lymphocytes preferentially cross the human BBB and accumulate in the CNS of mice during the effector phase of EAE. INTERPRETATION: Our data underscore the involvement of IFN-gamma(+) T(H)17 lymphocytes in the pathology of MS and EAE and their preferential recruitment into the CNS during inflammatory events. PMID- 19810098 TI - Titanium uptake, induction of RANK-L expression, and enhanced proliferation of human T-lymphocytes. AB - There is increasing evidence that titanium ions are released from orthopedic implants by biocorrosion. The aim of this study was to investigate titanium uptake by human T-lymphocytes and its effects on phenotype and proliferation. Freshly isolated human nonadherent peripheral blood mononuclear cells (NA-PBMC), were exposed to TiCl4 [Ti(IV)]. Bioavailability and distribution of Ti(IV) in T lymphocytes was determined by energy-filtered electron microscopy (EFTEM). The effects of Ti(IV) challenge on nonactivated and PHA-activated cells were assessed by flow cytometric analysis of surface markers, RANK-L production, and proliferation assays. EFTEM colocalized Ti(IV) with phosphorus in the nucleus, ribosomes, cytoplasmic membranes, and the surface membrane of T-lymphocytes. Ti(IV) increased significantly the expression of CD69, CCR4, and RANK-L in a concentration-dependent manner. Titanium enters T-lymphocytes through a currently unknown mechanism and binds to phosphorus-rich cell structures. Titanium influences phenotype and function of T-lymphocytes, resulting in activation of a CD69+ and CCR4+ T-lymphocyte population and secretion of RANK-L. These results strongly suggest the involvement of titanium ions challenged T-lymphocytes in the complex pathophysiological mechanisms of aseptic loosening of orthopedic implants. PMID- 19810099 TI - Serum levels of IGF-I, IGFBP-3 and colorectal cancer risk: results from the EPIC cohort, plus a meta-analysis of prospective studies. AB - Several prospective studies have shown a moderate positive association between increasing circulating insulin-like growth factor-I (IGF-I) levels and colorectal cancer risk. However, the associations were often statistically nonsignificant, and the relationship of cancer risk with IGF-I's major binding protein, IGFBP-3, showed major discrepancies between studies. We investigated the association of colorectal cancer risk with serum IGF-I, total and intact IGFBP-3, in a case control study nested within the EPIC cohort (1,121 cases of colorectal cancer and 1,121 matched controls). Conditional logistic regression was used to adjust for possible confounders. Our present study results were combined in a meta-analysis with those from 9 previous prospective studies to examine the overall evidence for a relationship of prediagnostic serum IGF-I with colorectal cancer risk. In the EPIC study, serum concentrations of IGF-I and IGFBP-3 showed no associations with risk of colorectal cancer overall. Only in subgroup analyses did our study show moderate positive associations of IGF-I levels with risk, either among younger participants only (and only for colon cancer) or among participants whose milk intakes were in the lowest tertile of the population distribution (RR for an increase of 100 ng/ml = 1.43 [95% CI = 1.13-1.93]). Nevertheless, in the meta analysis a modest positive association remained between serum IGF-I and colorectal cancer risk overall (RR = 1.07 [1.01-1.14] for 1 standard deviation increase in IGF-I). Overall, data from our present study and previous prospective studies combined indicate a relatively modest association of colorectal cancer risk with serum IGF-I. PMID- 19810100 TI - The dual PI3K/mTOR inhibitor PI-103 promotes immunosuppression, in vivo tumor growth and increases survival of sorafenib-treated melanoma cells. AB - Melanoma is the most lethal human skin cancer. If metastatic, it becomes very aggressive and resistant to standard modalities of anticancer treatment. During the last 10 years, several therapeutic strategies have been tested including the use of single and combined small drugs. Experimental results indicate that RAS and PI3K pathways are important for the development and maintenance of melanoma. In this study, we assessed the in vitro and in vivo inhibition potential of PI 103, a PI3K (p110alpha)/mTOR inhibitor and sorafenib, a BRAF inhibitor, as single agents and in combination in primary melanoma cell lines. Although PI-103 and sorafenib inhibited melanoma in vitro cell proliferation and viability, the inhibition of RAS pathway appeared to be more effective. The combination of the two agents in in vitro showed a synergistic effect inhibiting RAS and PI3K pathways in a cell line dependent manner. However, no cooperative effect was observed in blocking in vivo tumor growth in immunocompetent mice. In contrary to the expected, the data indicate that PI-103 induced immunosuppression promoting in vivo tumor growth and inhibiting apoptosis. Furthermore, in vitro studies examining the effects of the PI3K/mTOR inhibitor in tumor derived cell lines indicated that PI-103 induced the anti-apoptotic BH3 family proteins Mcl1, Bcl2 and Bcl(xL) favoring, the in vitro survival of sorafenib treated melanoma cells. These data certainly makes an argument for investigating unexpected effects of rational drug combinations on immunocompetent animal models prior to conducting clinical studies. PMID- 19810101 TI - Trimodal age-specific incidence patterns for Burkitt lymphoma in the United States, 1973-2005. AB - Burkitt lymphoma (BL) is a unique B-cell non-Hodgkin lymphoma with 3 established clinical-epidemiological variants: endemic, sporadic and AIDS-related BL. BL variants show characteristic dysregulation of MYC gene, but the causes of MYC dysregulation or BL arising at different ages are poorly understood. Therefore, we examined population-based BL incidence patterns in the United States to determine age-related risk. BL case and population data were obtained from the NCI's Surveillance, Epidemiology and End Results Databases (1973-2005). Standard cross-sectional age-standardized and age-specific incidence rates were stratified by sex and race and supplemented with age-period-cohort models. We analyzed 3,058 BL cases diagnosed during 1,160,300,297 person-years of observation. Age standardized incidence rates rose 6.8% per year (95% CI 4.5-9.1) for males and 7.1% (95% CI 3.2-11.1) for females during the study period. The rate among males was 3.2 times that among females, and among Whites 1.3 times that among Blacks. Male-to-female incidence rate ratios did not differ by race, but were 4.2 for pediatric (0-19 years), 4.1 for adult (20-59 years) and 2.0 for geriatric (> or = 60 years) BL. Cross-sectional age-specific rates showed 2 separate peaks among males and females, near ages 10 and 75 years, and a 3rd peak near age 40 years among males. The tri/bimodal incidence pattern was present in sensitivity analyses excluding registries with many HIV/AIDS cases and in period-specific, cohort-specific analyses. To our knowledge, tri/bimodal incidence patterns have not previously been reported for BL. Trimodal/bimodal BL suggests heterogeneity in etiology or biology of BL diagnosed at different ages in males and females. PMID- 19810102 TI - Skin cancers associated with HIV infection and solid-organ transplantation among elderly adults. AB - Immunosuppression may be etiologic for some skin cancers. We investigated the impact of human immunodeficiency virus (HIV) infection and solid-organ transplantation on skin cancer risk. We conducted a population-based case-control study among elderly U.S. adults (non-Hispanic whites, age 67 years or older), using Surveillance, Epidemiology and End Results Medicare linked data. The study comprised 29,926 skin cancer cases (excluding basal cell and squamous cell carcinomas) and 119,704 controls, frequency-matched by gender, age and calendar year (1987-2002). Medicare claims identified solid-organ transplantation or HIV infection before cancer diagnosis/control selection. As negative controls, we evaluated other medical conditions (e.g., hypertension and depression) and cancers (breast, colon and prostate) not linked to immunosuppression. Odds ratios (ORs) compared prevalence in cases and controls, adjusted for matching factors and number of prior physician claims. Risks of Kaposi sarcoma (N = 602) and cutaneous non-Hodgkin lymphoma (N = 1,836) were increased with solid-organ transplantation (OR [95%CI]: 11.06 [5.27-23.23] and 2.27 [1.00-5.15], respectively) and HIV infection (21.58 [11.94-38.99] and 2.41 [1.05-5.52], respectively). Solid-organ transplantation was also associated with increased risks of Merkel cell carcinoma (N = 1,286; OR [95%CI] 4.95 [2.62-9.34]) and other cutaneous sarcomas (N = 972; 4.19 [1.83-9.56]). Solid-organ transplantation was nonsignificantly associated with melanoma (N = 23,974; (OR 1.36 [95%CI 0.98 1.88]). Null or weak associations were observed for negative control medical conditions and cancers. Solid-organ transplantation and HIV infection were followed by increased risk for some skin cancer subtypes among elderly adults. These results highlight the potential role of immunity in development of skin cancers. PMID- 19810103 TI - Resveratrol enhances p53 acetylation and apoptosis in prostate cancer by inhibiting MTA1/NuRD complex. AB - Dietary compounds and epigenetic influences are well recognized factors in cancer progression. Resveratrol (Res), a dietary compound from grapes, has anticancer properties; however, its epigenetic effects are understudied. Metastasis associated protein 1 (MTA1) is a part of the nucleosome remodeling deacetylation (NuRD) corepressor complex that mediates posttranslational modifications of histones and nonhistone proteins resulting in transcriptional repression. MTA1 overexpression in prostate cancer (PCa) correlates with tumor aggressiveness and metastasis. In this study, we have identified a novel MTA1-mediated mechanism, by which Res restores p53-signaling pathways in PCa cells. We show, for the first time, that Res causes down-regulation of MTA1 protein, leading to destabilization of MTA1/NuRD thus allowing acetylation/activation of p53. We demonstrated that MTA1 decrease by Res was concomitant with accumulation of Ac-p53. MTA1 knockdown further sensitized PCa cells to Res-dependent p53 acetylation and recruitment to the p21 and Bax promoters. Furthermore, MTA1 silencing maximized the levels of Res-induced apoptosis and pro-apoptotic Bax accumulation. HDAC inhibitor SAHA, like MTA1 silencing, increased Res-dependent p53 acetylation and showed cooperative effect on apoptosis. Our results indicate a novel epigenetic mechanism that contributes to Res anticancer activities: the inhibition of MTA1/NuRD complexes due to MTA1 decrease, which suppresses its deacetylation function and allows p53 acetylation and subsequent activation of pro-apoptotic genes. Our study identifies MTA1 as a new molecular target of Res that may have important clinical applications for PCa chemoprevention and therapy, and points to the combination of Res with HDAC inhibitors as an innovative therapeutic strategy for the treatment of PCa. PMID- 19810104 TI - Effects of cyclic dynamic tensile strain on previously compressed inner annulus fibrosus and nucleus pulposus cells of human intervertebral disc-an in vitro study. AB - Our objective was to investigate whether dynamic tensile strain on previously compressed human intervertebral disc (IVD) cells can restore the biosynthetic effects of collagen and glycosaminoglycan. Inner annulus fibrosus (AF) and nucleus pulposus (NP) tissues of adolescent idiopathic scoliosis cases undergoing thoracoscopic discectomy and fusion were cultured on compressive plates. Compressive stress was applied using 0.4 MPa at 1 Hz, for 2 h twice a day for 7 days, to the inner AF and NP tissues, followed by equibiaxial cyclic tensile strain to deform the released cells onto the plate's flexible bottom. With 10% elongation at a rate of 1 Hz, for 2 h twice a day for 7 days, a significant increase in the level of collagen and glycosaminoglycan of the previously compressed inner AF, as well as the level of glycosaminoglycan of the previously compressed NP cells were found. The DNA content and number of endoplasmic reticulum under transmission electron micrograph of the previously compressed inner AF and NP cell were also significantly increased. The results suggested that equibiaxial cyclic tensile strain at a rate of 1 Hz with 10% tensile strain was capable of increasing collagen and glycosaminoglycan synthesis of previously compressed inner AF cells, and glycosaminoglycan synthesis of previously compressed NP cells. PMID- 19810105 TI - Low-density lipoprotein receptor-related protein 5 (LRP5) expression in human osteoarthritic chondrocytes. AB - The aim of this study was to investigate the activation of the Wnt/beta-catenin pathway in osteoarthritis and the role of low-density lipoprotein receptor related protein 5 (LRP5) in human osteoarthritic chondrocytes. The influence of 1,25(OH)2D3 on the expression of the LRP5 gene in human chondrocytes was also assessed. Human cartilage was obtained from 11 patients with primary osteoarthritis (OA) undergoing total knee replacement surgery. Normal cartilage was obtained from five healthy individuals. Beta-catenin and LRP5 mRNA levels were investigated using real-time PCR and LRP5 protein expression using Western blot analysis. Furthermore, we evaluated the effect of 1,25(OH)2D3 on LRP5 mRNA expression levels in osteoarthritic chondrocytes. Blocking LRP5 expression was performed using small interfering RNA (siRNA) against LRP5, and subsequent MMP-13 mRNA and protein levels were evaluated by real-time PCR and Western blot analysis, respectively. We confirmed the activation of the Wnt/beta-catenin pathway in OA, as we observed significant up-regulation of beta-catenin mRNA expression in osteoarthritic chondrocytes. We also observed that LRP5 mRNA and protein expression were significantly up-regulated in osteoarthritic cartilage compared to normal cartilage, and LRP5 mRNA expression was further increased by vitamin D. Also, blocking LRP5 expression using siRNA against LRP5 resulted in a significant decrease in MMP-13 mRNA and protein expressions. Our findings suggest the catabolic role of LRP5 is mediated by the Wnt/beta-catenin pathway in human osteoarthritis. PMID- 19810106 TI - Low-intensity pulsed ultrasound (LIPUS) increases the articular cartilage type II collagen in a rat osteoarthritis model. AB - In this study, the effect of low-intensity pulsed ultrasound (LIPUS) on cartilage was evaluated in a rat osteoarthritis (OA) model using serum biomarkers such as CTX-II (type II collagen degradation) and CPII (type II collagen synthesis) as well as histological criteria (Mankin score and immunohistochemical type II collagen staining). OA was surgically induced in the knee joint of rats by anterior cruciate/medial collateral ligament transection and medial meniscus resection (ACLT + MMx). Animals were divided into three groups: sham-operated group (Sham), ACLT + MMx group without LIPUS (-LIPUS), and ACLT + MMx group with LIPUS (+LIPUS; 30 mW/cm(2), 20 min/day for 28 days). CTX-II levels were elevated in both -LIPUS and +LIPUS groups compared to that in the Sham group after the operation, but there was no significant difference between +LIPUS and -LIPUS groups, suggesting that LIPUS does not affect the degradation of type II collagen in this model. In contrast, CPII was significantly increased in +LIPUS group compared to -LIPUS and Sham. Moreover, histological damage on the cartilage (Mankin score) was ameliorated by LIPUS, and type II collagen was immunohistochemically increased by LIPUS in the cartilage of an OA model. Of interest, mRNA expression of type II collagen was enhanced by LIPUS in chondrocytes. Together these observations suggest that LIPUS is likely to increase the type II collagen synthesis in articular cartilage, possibly via the activation of chondrocytes and induction of type II collagen mRNA expression, thereby exhibiting chondroprotective action in a rat OA model. PMID- 19810107 TI - Occupational exposures contribute to educational inequalities in lung cancer incidence among men: Evidence from the EPIC prospective cohort study. AB - The aim of this study is to investigate to what extent occupational exposures may explain socioeconomic inequalities in lung cancer incidence after adjusting for smoking and dietary factors. Analyses were based on a subsample of the European Prospective Investigation into Cancer and Nutrition (EPIC study), a prospective cohort. The analyses included 703 incident lung cancer cases among men in Denmark, the United Kingdom, Germany, Italy, Spain and Greece. The socioeconomic position was measured using the highest level of education. The estimates of relative indices of inequality (RII) were computed with Cox regression models. We first adjusted for smoking (with detailed information on duration and quantity) and dietary factors (fruits and vegetables consumption) and then for occupational exposures. The exposure to three carcinogens [asbestos, heavy metals and polycyclic aromatic hydrocarbons (PAH)] was analyzed. The occupational exposures explained 14% of the socioeconomic inequalities remaining after adjustment for smoking and fruits and vegetables consumption. The inequalities remained nevertheless statistically significant. The RII decreased from 1.87 (95% CI: 1.36 2.56) to 1.75 (1.27-2.41). The decrease was more pronounced when adjusting for asbestos than for heavy metals or PAH. Analyses by birth cohort suggested an effect of occupational exposures among older men, while due to small number of endpoints, no conclusion could be drawn about the role of occupational exposures in educational inequalities among younger men. Our study revealed that the impact of occupational exposures on socioeconomic inequalities in cancer incidence, rarely studied until now, exists while of modest magnitude. PMID- 19810108 TI - Structural stability and aggregation behavior of the VEALYL peptide derived from human insulin: a molecular dynamics simulation study. AB - The VEALYL peptide from B chain (residues 12-17) of insulin has been shown to form amyloid-like fibrils. Recently, the atomic structure of the VEALYL oligomer has been determined by X-ray microcrystallography and reveals a dry, tightly self complementing structure between the neighboring beta-sheet layers, termed as "steric zipper." In this study, several molecular dynamics simulations with all atom explicit water were conducted to investigate the structural stability and aggregation behavior of the VEALYL peptide with various sizes and its single glycine replacement mutations. The results of our single-layer models showed that the structural stability of the VEALYL oligomers increases significantly with increasing the number of beta-strands. We further suggested that the minimal nucleus seed for VEALYL fibril formation could be as small as three or four peptides. Our results also revealed that the hydrophobic interaction between E2 and Y5 plays an important role in stabilizing the adjacent beta-strands within the same layer, whereas the hydrophobic steric zipper formed via the side chains of V1, A3, L4, Y5, and L6 locks the two neighboring beta-sheet layers together. Mutation simulations showed that the substitution of a single glycine residue directly disrupts this steric zipper, resulting in the destabilization of the VEALYL oligomers. This study provides the atomic insights into understanding the aggregation behavior of the VEALYL peptide. It may also be helpful for designing new or modified capping peptides able to break the driving force for aggregation and to prevent the fibril formation of the VEALYL peptide and the insulin protein. PMID- 19810109 TI - Thermal stability and conformational structure of salmon calcitonin in the solid and liquid states. AB - Salmon calcitonin (sCT) was selected as a model protein drug for investigating its intrinsic thermal stability and conformational structure in the solid and liquid states by using a Fourier transform infrared (FT-IR) microspectroscopy with or without utilizing thermal analyzer. The spectral correlation coefficient (r) analysis between two second-derivative IR spectra was applied to quantitatively estimate the structural similarity of sCT in the solid state before and after different treatments. The thermal FT-IR microspectroscopic data clearly evidenced that sCT in the solid state was not effected by temperature and had a thermal reversible property during heating-cooling process. Moreover, the high r value of 0.973 or 0.988 also evidenced the structural similarity of solid state sCT samples before and after treatments. However, sCT in H(2)O exhibited protein instability and thermal irreversibility after incubation at 40 degrees C. The temperature-induced conformational changes of sCT in H(2)O was occurred to transform the alpha-helix/random coil structures to beta-sheet structure and also resulted in the formation of intramolecular and intermolecular beta-sheet structures. PMID- 19810110 TI - Neomycin fixation followed by ethanol pretreatment leads to reduced buckling and inhibition of calcification in bioprosthetic valves. AB - Glutaraldehyde crosslinked bioprosthetic heart valves (BHVs) have two modalities of failure: degeneration (cuspal tear due to matrix failure) and calcification. They can occur independently as well as one can lead to the other causing co existence. Calcific failure has been extensively studied before and several anti calcification treatments have been developed; however, little research is directed to understand mechanisms of valvular degeneration. One of the shortcomings of glutaraldehyde fixation is its inability to stabilize all extracellular matrix components in the tissue. Previous studies from our lab have demonstrated that neomycin could be used as a fixative to stabilize glycosaminoglycans (GAGs) present in the valve to improve matrix properties. But neomycin fixation did not prevent cuspal calcification. In the present study, we wanted to enhance the anti-calcification potential of neomycin fixed valves by pre-treating with ethanol or removing the free aldehydes by sodium borohydride treatment. Ethanol treatment has been previously used and found to have excellent anti-calcification properties for valve cusps. Results demonstrated in this study suggest that neomycin followed by ethanol treatment effectively preserves GAGs both in vitro as well as in vivo after subdermal implantation in rats. In vivo calcification was inhibited in neomycin fixed cusps pretreated with ethanol compared to glutaraldehyde (GLUT) control. Sodium borohydride treatment by itself did not inhibit calcification nor stabilized GAGs against enzymatic degradation. Neomycin fixation followed by ethanol treatment of BHVs could prevent both modalities of failure, thereby increasing the effective durability and lifetime of these bioprostheses several fold. PMID- 19810111 TI - Selective laser melting: a unit cell approach for the manufacture of porous, titanium, bone in-growth constructs, suitable for orthopedic applications. II. Randomized structures. AB - In this study, the unit cell approach, which has previously been demonstrated as a method of manufacturing porous components suitable for use as orthopedic implants, has been further developed to include randomized structures. These random structures may aid the bone in-growth process because of their similarity in appearance to trabecular bone and are shown to carry legacy properties that can be related back to the original unit cell on which they are ultimately based. In addition to this, it has been shown that randomization improves the mechanical properties of regular unit cell structures, resulting in anticipated improvements to both implant functionality and longevity. The study also evaluates the effect that a post process sinter cycle has on the components, outlines the improved mechanical properties that are attainable, and also the changes in both the macro and microstructure that occur. PMID- 19810112 TI - Effect of low level capacitive-coupled pulsed electric field stimulation on mineral profile of weight-bearing bones in ovariectomized rats. AB - Mineral content, mineral composition, and crystalline pattern of bone in osteoporosis are different from those of normal individuals. Present management of bone mineral loss is rather unsatisfactory primarily because of socioeconomic factors and untoward effects of the treatment drugs. We report the efficacy of capacitive-coupled pulsed electric field (CCPEF) to prevent bone loss in an ovariectomized rat model of osteoporosis. One month postsurgery either leg was stimulated with CCPEF, whereas the other leg did not receive any stimulation (sham exposed). The treatment was given in 60 sessions each of 2 h/d (5 days a week). The control group of rats was sham operated. At the end of the observation period, femur and tibia bones were removed. Their bone mineral content (BMC), calcium, phosphorus, and carbon contents were analyzed and bone mineral density (BMD) was calculated. The BMC data were supported by X-ray diffraction (XRD) method. In sham-exposed bones, a statistically significant decrease in BMC, BMD, calcium, and phosphorus contents were obtained as compared to the control. Although in CCPEF bones, there was an attenuation of decrement in the noted parameters except phosphorus. XRD pattern supported these observations. The results suggest that chronic, 60 sessions of 2 h/d, 5 d/wk CCPEF (14 MHz with 16 Hz modulation 16 Hz and 10 V peak to peak) is effective in attenuating the ovariectomy-induced bone mineral loss in rats. PMID- 19810113 TI - Oral bacterial adhesion on amorphous carbon and titanium films: effect of surface roughness and culture media. AB - Implant infections can cause severe problems from malfunctioning to dangerous sepsis affecting the health of the patient. For many years, titanium has been the most common material used on dental implants due to their mechanical and biocompatibility properties. Recent studies suggest that amorphous carbon (a-C) films can be possible candidates for coating dental implants, improving some important features like biocompatibility and bone formation. In the oral cavity, the risk of an implant infection is high due to multiple species are capable to colonize this site and these biofilm infections can limit the use of these medical devices. The purpose of this study was to evaluate the influence of the surface chemistry, roughness, and culture media in the bacterial colonization process. To achieve this, a-C and Ti films were deposited on rough and smooth surfaces and cultured with different microorganisms belonging to the oral microbiota with mycoplasma medium (MM) or human saliva (HS). Samples were incubated for 24 h, after this, samples were sonicated and the number of attached bacteria was determined by counting the colony-forming units (CFU's) from each sample. The proportion of the species in the biofilms was determined using checkerboard DNA-DNA hybridization. Data were analyzed by Student's t test using Bonferroni's modification of Student's t test and differences on the proportion of the bacterial species attached to each surface were determined using the Mann Whitney test. Results show an increased number of CFU's on rough surfaces, especially on the a-C surfaces. The incubation media were an important factor on the adhesion of certain taxa, whereas other species were more sensitive to surface chemistry and others to surface roughness. PMID- 19810114 TI - Can bone healing in distraction osteogenesis be accelerated by local application of IGF-1 and TGF-beta1? AB - Because complications of distraction osteogenesis are largely related to the long duration of therapy, increasing efforts were reached to shorten treatment by using osteoconductive replacement materials incorporating bioactive molecules such as IGF-1 and TGF-beta1. The controlled release of IGF-1 and TGF-beta1 from coated biodegradable poly(D,L-lactide) implants could stimulate fracture healing locally. We investigated the effect of locally applied IGF-1 and TGF-beta1 from IGF-1/TGF-beta1-enriched polylactide membranes on fracture healing in a sheep model of delayed callus formation. Twenty-eight sheep were used for this study. Callus distraction of 1 mm/day by means of a unilateral fixator was continued for 30 days. At the beginning of the subsequent consolidation phase, either growth factors were applied locally or the defect was packed with cancellous bone, or both. The groups treated with growth factors were compared to a control group. The consolidation phase lasted for 60 days and both tibiae were dissected for histological and histomorphometric analyses. This investigation found a reduced absolute callus area in the lengthening zone in all treatment groups. The two treatment groups that received a membrane coated with growth factors showed distinctly higher relative bone areas than the groups treated with an uncoated membrane or packing of the osteotomy defect with cancellous bone. The differences in bone areas were not statistically significant. Application of the growth factors accelerated bone healing and achieved results comparable with those of established treatment methods (packing with autologous cancellous bone). The best results were achieved with a combination of both methods. PMID- 19810115 TI - Preparation and characterization of PMAA/MWCNTs nanohybrid hydrogels with improved mechanical properties. AB - A novel nanohybrid hydrogel was in situ prepared by means of a free radical crosslinking polymerization route of methacrylic acid in the presence of multi walled carbon nanotubes (MWCNTs). The structural and morphological characterizations revealed that poly(methacrylic acid) networks (PMAA) closely covered the MWCNTs, and a MWCNT-well-dispersed nanohybrid hydrogel was formed. The addition of MWCNTs strikingly improved pH response and mechanical properties, depending on the component ratios and particle sizes of MWCNTs as well as crosslinker concentrations. The swelling rate was obviously faster than that of the pure PMAA hydrogel. The hydrophilic nature of polyelectrolytes, the capillarity effect, cation-pi or charge-transfer interaction and hydrogen bonds, as well as a subtle balance among these interactions were adopted to interpret the above swelling behavior. Load transfer to the MWCNTs in the networks played important part in compression mechanical improvements. MTT assays were adopted to evaluate the cytocompatibility of the developed biomaterials. This smart hydrogel is expected to be used as potential candidate for specific biological applications. PMID- 19810116 TI - Corono-apical leakage of various root filling materials using two different penetration models--a 3-month study. AB - The purpose of this study was to analyze the sealing ability of various filling combinations in a 90 days period using fluid transport and glucose penetration models. Premolars (n = 108) were filled with either gutta-percha (groups 1-4) or Resilon (groups 5-8) core materials combined with AH Plus (groups 1, 3, 5, 7) or Epiphany (groups 2, 4, 6, 8) sealers by using cold lateral compaction (groups 1, 2, groups 5, 6) or System B with Obtura II (groups 3, 4, groups 7, 8). The sealing ability of each sample was evaluated through glucose and fluid penetration models. Penetration values of 90 days were statistically analyzed with three-way analysis of variance (ANOVA)and Duncan's Multiple Range tests (p < 0.05). No significant differences were found among the penetration values when the fluid filtration model was used (p > 0.05). On the contrary, statistically significant differences were obtained among the groups with the glucose penetration model (p < 0.05). AH Plus/Resilon/cold lateral compaction group had better sealing ability than the other combinations tested with the glucose penetration model. PMID- 19810117 TI - Effect of adipic dihydrazide modification on the performance of collagen/hyaluronic acid scaffold. AB - Collagen and hydrazide-functionalized hyaluronic acid derivatives were hybridized by gelating and genipin crosslinking to form composite hydrogel. The study contributed to the understanding of the effects of adipic dihydrazide modification on the physicochemical and biological properties of the collagen/hyaluronic acid scaffold. The investigation included morphology observation, mechanical measurement, swelling evaluation, and collagenase degradation. The results revealed that the stability of composites was increased through adipic dihydrazide modification and genipin crosslinking. The improved biocompatibility and retention of hyaluronic acid made the composite material more favorable to chondrocytes growing, suggesting the prepared scaffold might be high potential for chondrogenesis. PMID- 19810118 TI - Dental glass-reinforced composite for caries inhibition: calcium phosphate ion release and mechanical properties. AB - The two main challenges facing dental composite restorations are secondary caries and bulk fracture. Previous studies developed whisker-reinforced Ca-PO(4) composites that were relatively opaque. The objective of this study was to develop an esthetic glass particle-reinforced, photo-cured calcium phosphate composite. Tetracalcium phosphate (TTCP) particles were incorporated into a resin for Ca and PO(4) release, while glass particles provided reinforcement. Ion release and mechanical properties were measured after immersion in solutions with pH of 7, 5.5, and 4. For the composite containing 40% mass fraction of TTCP, incorporating glass fillers increased the strength (p < 0.05). Flexural strength (Mean +/- SD; n = 6) at 30% glass was 99 +/- 18 MPa, higher than 54 +/- 20 MPa at 0% glass (p < 0.05). Elastic modulus was 11 GPa at 30% glass, compared to 2 GPa without glass. At 28 days, the released Ca ion concentration was 4.61 +/- 0.18 mmol/L at pH of 4, much higher than 1.14 +/- 0.07 at pH of 5.5, and 0.27 +/- 0.01 at pH of 7 (p < 0.05). PO(4) release was also dramatically increased at cariogenic, acidic pH. The TTCP-glass composite had strength 2-3 fold that of a resin-modified glass ionomer control. In conclusion, the photo-cured TTCP-glass composite was "smart" and substantially increased the Ca and PO(4) release when the pH was reduced from neutral to a cariogenic pH of 4, when these ions are most needed to inhibit tooth caries. Its mechanical properties were significantly higher than previous Ca, PO(4), and fluoride releasing restoratives. Hence, the photo-cured TTCP-glass composite may have potential to provide the necessary combination of load-bearing and caries-inhibiting capabilities. PMID- 19810119 TI - Widening the mutation spectrum of EVC and EVC2: ectopic expression of Weyer variants in NIH 3T3 fibroblasts disrupts Hedgehog signaling. AB - Autosomal recessive Ellis-van Creveld syndrome and autosomal dominant Weyer acrodental dysostosis are allelic conditions caused by mutations in EVC or EVC2. We performed a mutation screening study in 36 EvC cases and 3 cases of Weyer acrodental dysostosis, and identified pathogenic changes either in EVC or in EVC2 in all cases. We detected 40 independent EVC/EVC2 mutations of which 29 were novel changes in Ellis-van Creveld cases and 2 were novel mutations identified in Weyer pedigrees. Of interest one EvC patient had a T>G nucleotide substitution in intron 7 of EVC (c.940-150T>G), which creates a new donor splice site and results in the inclusion of a new exon. The T>G substitution is at nucleotide +5 of the novel 5' splice site. The three Weyer mutations occurred in the final exon of EVC2 (exon 22), suggesting that specific residues encoded by this exon are a key part of the protein. Using murine versions of EVC2 exon 22 mutations we demonstrate that the expression of a Weyer variant, but not the expression of a truncated protein that mimics an Ellis-van Creveld syndrome mutation, impairs Hedgehog signal transduction in NIH 3T3 cells in keeping with its dominant effect. PMID- 19810120 TI - Multiple osteochondromas: mutation update and description of the multiple osteochondromas mutation database (MOdb). AB - Multiple osteochondromas (MO) is an autosomal dominant skeletal disease characterized by the formation of multiple cartilage-capped bone tumors growing outward from the metaphyses of long tubular bones. MO is genetically heterogeneous, and is associated with mutations in Exostosin-1 (EXT1) or Exostosin-2 (EXT2), both tumor-suppressor genes of the EXT gene family. All members of this multigene family encode glycosyltransferases involved in the adhesion and/or polymerization of heparin sulfate (HS) chains at HS proteoglycans (HSPGs). HSPGs have been shown to play a role in the diffusion of Ihh, thereby regulating chondrocyte proliferation and differentiation. EXT1 is located at 8q24.11-q24.13, and comprises 11 exons, whereas the 16 exon EXT2 is located at 11p12-p11. To date, an EXT1 or EXT2 mutation is detected in 70-95% of affected individuals. EXT1 mutations are detected in +/-65% of cases, versus +/-35% EXT2 mutations in MO patient cohorts. Inactivating mutations (nonsense, frame shift, and splice-site mutations) represent the majority of MO causing mutations (75 80%). In this article, the clinical aspects and molecular genetics of EXT1 and EXT2 are reviewed together with 895 variants in MO patients. An overview of the reported variants is provided by the online Multiple Osteochondromas Mutation Database (http://medgen.ua.ac.be/LOVD). PMID- 19810121 TI - Student laboratory presentations as a learning tool in anatomy education. AB - Previous studies have shown that anatomy students who complete oral laboratory presentations believe they understand the material better and retain it longer than they otherwise would if they only took examinations on the material; however, we have found no studies that empirically test such outcomes. The purpose of this study was to assess the effectiveness of oral presentations through comparisons with other methods of assessment, most notably, examination performance. Specifically, we tested whether students (n = 256) performed better on examination questions on topics covered by their oral presentations than on other topics. Each student completed two graded, 12-minute laboratory presentations on two different assigned topics during the course and took three examinations, each of which covered a third of the course material. Examination questions were characterized by type (memorization, pathway, analytical, spatial). A two-way repeated measures analysis of variance revealed that students performed better on topics covered by their presentations than on topics not covered by their presentations (P < 0.005), regardless of presentation grade (P > 0.05) and question type (P > 0.05). These results demonstrate empirically that oral presentations are an effective learning tool. PMID- 19810122 TI - A second-generation ionic liquid matrix-assisted laser desorption/ionization matrix for effective mass spectrometric analysis of biodegradable polymers. AB - A second generation ionic liquid matrix (ILM), N,N-diisopropylethylammonium alpha cyano-4-hydroxycinnamate (DEA-CHCA), was developed for the characterization of polar biodegradable polymers. It is compared with five solid matrices typically used for the characterization of these polymers and one other new ILM. It is shown that use of the ILM, DEA-CHCA, allows maximum signal with minimum laser intensity which minimizes polymer degradation. In these conditions, the DEA-CHCA ILM is able to assist in the ionization of analytes in an efficient but soft manner. These qualities produce spectra that allow an accurate and sensitive determination of the number average molecular weights, weight average m.w., and polydispersity index of labile polar polymers. With such polymers, many solid matrices produce spectra showing extensive polymer degradation leading to the underestimation of molecular weights. The distribution of intact analyte peaks obtained with the ILM DEA-CHCA allows for identification of the fine structure of complex copolymers. ILMs were much less susceptible to effects of extraction delay times on molecular weight determination than were solid matrices. The liquid nature of the matrix is an important reason for the outstanding results obtained for labile analyte polymers. No comparable results could be obtained with any known solid matrices or other ILMs. In many cases, the manufacturers' listed molecular weights and polydispersity measurements for biodegradable polymers are determined by size-exclusion chromatography and the data obtained by that method may differ considerably from the high-precision matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) results presented here. PMID- 19810123 TI - Analysis of sesquiterpene lactones in Eupatorium lindleyanum by HPLC-PDA-ESI MS/MS. AB - INTRODUCTION: The aerial part Eupatorium lindleyanum is commonly used as an antipyretic and detoxicant clinically in traditional Chinese medicine. Our previous research showed that germacrane sesquiterpene lactones were its main active constituents, so the development of rapid and accurate methods for the identification of the sesquiterpene lactones is of great significance. OBJECTIVE: To develop an HPLC-PDA-ESI-MS/MS method capable for simple and rapid analysis of germacrane sesquiterpene lactones in the aerial part E. lindleyanum. METHODOLOGY: High-performance liquid chromatography-photodiode array detection-electrospray ionization-tandem mass spectrometry was used to analyze germacrane sesquiterpene lactones of Eupatorium lindleyanum. The fragmentation behavior of germacrane sesquiterpene lactones in a Micromass Q/TOF Mass Spectrometer was discussed, and 9 germacrane sesquiterpene lactones were identified by comparison of their characteristic data of HPLC and MS analyses with those obtained from reference compounds. RESULTS: The investigated germacrane sesquiterpene lactones were identified as eupalinolides C (1), 3beta-acetoxy-8beta-(4'-hydroxy-tigloyloxy)-14 hydroxy-costunolide (2), eupalinolides A (3), eupalinolides B (4), eupalinolides E (5), 3beta-acetoxy-8beta-(4'-oxo-tigloyloxy)-14-hydroxy-heliangolide (6), 3beta acetoxy-8beta-(4'-oxo- tigloyloxy)-14-hydroxy-costunolide (7), hiyodorilactone B (8), and 3beta-acetoxy-8beta-(4'-hydroxy-tigloyloxy)- costunolide (9). Compounds 6, 7 and 9 were reported for the first time. CONCLUSION: HPLC-PDA-ESI-MS/MS provides a new powerful approach to identify germacrane sesquiterpene lactones in E. lindleyanum rapidly and accurately. PMID- 19810124 TI - Development and validation of a rapid capillary zone electrophoresis method for the determination of aconite alkaloids in aconite roots. AB - INTRODUCTION: Aconites, with aconite alkaloids as the major therapeutic and toxic components, are used for the treatment of analgesic, antirheumatic and neurological symptoms. Quantification of the aconite alkaloids is important for the quality control of aconite-containing drugs. OBJECTIVE: To establish a validated capillary zone electrophoresis (CZE) method for the simultaneous determination of six major alkaloids, namely aconitine, mesaconitine, hypaconitine, benzoylaconine, benzoylmesaconine and benzoylhypaconine, in crude and processed aconite roots. METHODOLOGY: The CZE method was optimised and validated using a stability-indicating method. The optimised running buffer was a mixture of 200 mm Tris, 150 mm perchloric acid and 40% 1,4-dioxane (pH 7.8) with the capillary thermostated at 25 degrees C. RESULTS: Using the optimised method, six aconite alkaloids were well separated. The established method showed good precision, accuracy and recovery. Contents of these alkaloids in crude and processed aconites were determined and it was observed that the levels of individual alkaloids varied between samples. CONCLUSION: The developed CZE method was reliable for the quality control of aconites contained in herbal medicines. The method could also be used as an approach for toxicological studies. PMID- 19810125 TI - Radiotherapy and chemotherapy in locally advanced head and neck squamous cell carcinoma. AB - Throughout the past two decades the efforts to improve the efficacy of treatment for locally advanced head and neck squamous cell carcinoma (HNSCC) have led to increased use of multimodality approaches combining surgery, radiotherapy (RT), and chemotherapy (CT). Conventional RT, a standard approach for locoregionally advanced disease, was associated with unsatisfactory results, thereby, a greater understanding of radiobiology led to the development of two classes of altered radiation fractionation schedules incorporating hyperfractionation (HF) and acceleration in the management of advanced HNSCC. Randomized controlled trials and meta-analyses demonstrated that for patients with locally advanced HNSCC major improvements in locoregional control (LRC) at high level of evidence can be achieved by accelerated fractionation (AF) and HF. For these patients, overall survival (OS) may be improved at high level of evidence by HF delivered with increased total dose. CT represents an important component of multimodality treatment approach for locally advanced HNSCC with concurrent addition of CT to RT being the most significant method for improving head and neck cancer (HNC) outcome. Several randomized studies and meta-analyses on the administration of concurrent chemoradiotherapy (CCRT) demonstrated clear evidence that CCRT provides a substantial and statistically significant improvement in survival and locoregional control, as compared to RT alone. CCRT is now a standard treatment approach for patients with locally advanced HNSCC. CCRT has been also shown to allow organ preservation in almost two thirds of patients without affecting survival. Recently, strong evidence for an improved outcome for high-risk resected patients has been shown by the use of adjuvant CCRT. PMID- 19810126 TI - Adjuvant therapy with aromatase inhibitors in postmenopausal, estrogen receptor- positive breast cancer patients: upfront or sequential? AB - For decades tamoxifen (TAM) has been the mainstay hormonal treatment for estrogen receptor positive (ER+) breast cancer patients. Nevertheless, during the last years, for postmenopausal women particularly, the third generation aromatase inhibitors (AI) became the preferred alternative. The results of the randomized trials showed that AI were superior to TAM in terms of efficacy, and were accompanied by a different but fairly convenient side effects profile. Subsequently, all updated guidelines recommend the use of AI in the adjuvant setting for this category of patients, either upfront, following 2-3 years of TAM or as extended adjuvant therapy, after 5 years of TAM. However, no consensus has been reached regarding the best strategy to be used, and the expert opinion is divided, based on the available evidence. The controversial aspect of whether AI should be used upfront or following 2-3 years of TAM is further detailed in this manuscript, and some useful recommendations are provided in order to facilitate the decision-making process during the current clinical practice. PMID- 19810127 TI - Non small cell lung cancer: surgical treatment in the elderly. AB - Lung cancer resection in elderly patients is justified and has decreasing morbidity and mortality rates. Careful patient selection and operative planning are necessary, however. It is wise to have a diagnosis and staging done before the patient arrives in the operating theater. The surgeon should avoid extended resections when possible. In addition, elderly patients should be ambulated as soon as possible and adequate pain control should be ensured. Finally, the stage of the disease and occurrence of cardiopulmonary complications are the main determinants of outcome. PMID- 19810128 TI - Nature of cervical cancer and other HPV - associated cancers. AB - Papillomaviruses are small DNA viruses that infect and multiply in cutaneous or mucosal epithelial tissue. Human papillomavirus (HPV) 16 and 18 cause more than 99% of cervical carcinomas. Simultaneous presence of HPV is found in cervical intraepithelial neoplasia, vaginal intraepithelial neoplasia, vaginal and anal cancer. Invasive vulvar squamous cell carcinoma in younger women under the age of 50 are also associated with HPV. Most of the penile lesions are subclinical and the high prevalence of high-risk HPV suggests that they constitute a reservoir for high-risk HPV. Bowens disease and Buschke-Lowenstein tumors are associated with particular low- and high-risk HPV types. The potential role of HPV infection in the carcinogenic steps of breast, prostate, colorectal and lung cancers should be further tested. HPV-DNA might be transported from the original site of infection to the breast tissue by the bloodstream, and therefore is possibly involved in the carcinogenesis of breast neoplasia in some patients. HPV-DNA is detected in 40-70% of head and neck squamous cell carcinomas and in only 1% in normal epithelial cells. In this paper we propose the hypothesis that many epithelial normal cells are susceptible to HPV infection, which are the most sexually transmitted viruses. Experimental and epidemiological data imply a causative role for HPVs and they appear to be the second most important risk factor for cancer development in humans, exceeded only by tobacco usage. PMID- 19810129 TI - New therapeutic approaches in the management of metastatic renal cell carcinoma. AB - Renal cell carcinoma (RCC) accounts for approximately 2% of all cancers worldwide. For several decades cytokine therapy using either interleukin-2 (IL-2) or interferon -alpha (IFN-alpha) was the only effective treatment available for patients with metastatic RCC. Metastatic RCC represents a chemotherapy- and radiotherapy-resistant tumor with poor prognosis and 5-year survival rate <10%. Over the past few years considerable advances in the understanding of biology and underlying molecular pathways in RCC have resulted in the development of targeted therapies for this disease. PMID- 19810130 TI - Laryngeal paragangliomas - controversies in diagnosis and management. AB - Paragangliomas of the larynx are rare neuroendocrine tumors with difficulties in diagnosis and management. A review of the literature was carried out, using Medline and other available databases. Electronic links and related books were also included. The most reliable diagnostic imaging procedures are MRI, CT scan, and octreotide scintigraphy. Complete surgical excision with the maximal possible preservation of the laryngeal function is the treatment of choice. Many different surgical techniques have been reported but open surgical procedures seem to provide better results with lower recurrence rates, although endoscopic approaches and laser surgery have also been used with variable results. The use of long-acting depot octreotide has been used for the relief of symptoms and stabilization of the disease, especially in inoperable lesions with positive initial octreotide scintigraphy imaging. Detailed preoperative assessment and treatment planning in individual basis are essential in the management of these tumors. PMID- 19810131 TI - Risk index score for bacteremia in febrile neutropenic episodes in children with malignancies. AB - PURPOSE: To prospectively determine risk factors for bacteremia in febrile neutropenic children with malignancies. PATIENTS AND METHODS: We studied 199 episodes of febrile neutropenia in 80 children with malignancies, treated by conventional chemotherapy for a 4-year period (2000 - 2004). A standardized computer database with a set of variables for each febrile neutropenic episode was used. C-reactive protein (CRP) was measured at the first febrile episode and on the 3rd and 5th day from the beginning of antibiotic therapy by immunoturbidimetric method. Blood cultures were taken at the onset of fever and before initiation of antibiotic therapy. RESULTS: Multivariate logistic regression analysis determined 5 variables as independent risk factors for bacteremia: the underlying malignant disease (leukemia, non - Hodgkin's lymphoma / NHL, stage IV), chills, perianal cellulitis, presence of central venous catheter and CRP rise >or=34.5 mg/L between the onset of fever and the 3rd day of empiric antibiotic therapy. Thus we identified a low-risk group for bacteremia of 19.1%. CONCLUSION: Serial measurement of CRP allows for definite risk stratification of febrile neutropenic episodes on the 3rd day from the onset of fever. The low-risk group could be eligible for sequential empiric antibiotic therapy. PMID- 19810132 TI - Malignant pleural effusion and talc pleurodesis. Experimental model regarding early kinetics of talc particle dissemination in the chest after experimental talc pleurodesis. AB - PURPOSE: Talc remains a commonly used agent for pleurodesis malignant pleural effusion. Nevertheless, it is associated with a 3-9% incidence of pulmonary reactions ranging from simple pneumonitis to acute respiratory distress syndrome (ARDS). The underlying lung pathology and the size and rate of talc particle dissemination have been implicated as the cause of these complications. There seems to be an acknowledged lack of evidence regarding detailed very early intrathoracic talc particle migration. MATERIALS AND METHODS: Thirty white male New Zealand rabbits underwent experimental pleurodesis and were randomly assigned to 3 (A, B, C) study groups (10 in each group). Rabbits were sacrificed 6, 12 and 18 h after talc administration. Samples from both lungs, mediastinum and parietal pleura were obtained. The number of talc crystals (m) deposited was counted and averaged along all slices of the various tissue samples. RESULTS: A high degree of early talc deposition and subsequent epithelial injury in all examined tissues was observed. Diffuse talc deposition occurred in both lungs, but in a different manner. On the side of talc administration, talc particles were deposited in a time-dependent fashion. On the contralateral side, talc was rapidly deposited during the first hours after the procedure, then the rate of deposition decreased, and increased again between 12 and 18 h after the procedure. CONCLUSION: Large-sized talc particles are deposited on both lungs very early after pleurodesis. At the same time inflammatory pulmonary changes appear bilaterally. Despite contradicting data in the literature, these findings should always be kept in mind when performing this procedure in high risk patients. PMID- 19810133 TI - Breast cancer in association with thyroid disorders. AB - PURPOSE: The relationship between breast cancer and thyroid diseases is controversial. Conflicting results have been reported in the literature. The incidence of autoimmune and non-autoimmune thyroid diseases were investigated in patients with breast cancer who had received prior therapy as compared with age matched control individuals without breast or thyroid disease. PATIENTS AND METHODS: Clinical and ultrasound evaluation of the thyroid gland, and determination of serum thyroid hormones and autoantibody levels were performed in 143 breast cancer patients and 128 healthy control individuals. Patients were classified into subgroups according to estrogen receptor (ER) and progesterone receptor (PR) status and type of oncological treatment. RESULTS: The mean values for serum antibodies against thyroid peroxidase (anti-TPO) were 9 IU/ml and 25 IU/ml for antithyroglobulin antibodies (anti-TGB) in breast cancer patients, and 9.5 IU/ml and 23.5 IU/ml, respectively, in the control group (p>0.05. The difference between breast cancer patients and the control group in the incidence of autoimmune and non-autoimmune thyroid diseases was not statistically significant. No significant differences between the groups according to both menopausal status and ER status were seen (p= 0.67). Also, no significant influence of hormonal therapy with tamoxifen and chemotherapy on serum levels of thyroid stimulating hormone (TSH), free thyroxin (fT4), TPO and TGB autoantibodies was proved. CONCLUSION: This study demonstrated a similar incidence of thyroid enlargement and the same frequency of thyroid disturbances in patients with breast cancer and controls. No relationship was found among ER and PR status, and the presence of serum thyroid autoantibodies. Although we have been unable to demonstrate any impact of breast cancer therapy on thyroid function tests, more prolonged studies with larger number of patients may be required to demonstrate significant trends. PMID- 19810134 TI - Efficacy and toxicity of lower dose UFT without leucovorin in metastatic gastric cancer patients. AB - PURPOSE: Gastric cancer is the 4th most commonly diagnosed cancer and the 2nd leading cause of cancer death worldwide. In this study assessed were the efficacy and toxicity of the combination of epirubicin, cisplatin and UFT in patients with metastatic gastric cancer (MGC). PATIENTS AND METHODS: In this retrospective study 27 patients with MGC were treated with epirubicin 50 mg/m(2) and cisplatin 60 mg/m(2) i.v. on day 1 and subsequently UFT 250 mg/m(2)/day orally in divided doses for 21 days, followed by a 7/day rest (EP/UFT). RESULTS: Response and toxicity evaluation was possible for 25 patients. Three complete (12%) and 2 partial (8%) responses were observed. With a median follow-up 37 weeks (range 15 117), the median progression-free survival (PFS) and overall survival (OS) were 24 and 31 weeks, respectively. WHO grade 3 or 4 toxicity included neutropenia in 3 (12%) patients and nausea/vomiting in 1 (4%) patient. Neutropenic fever developed in only 1 (4%) patient. CONCLUSION: EP-UFT with lower UFT doses and without leucovorin support is a safe and effective regimen as first -line treatment of MGC. PMID- 19810135 TI - Profile of handicap situations in cancer patients. AB - PURPOSE: The aim of this study was to assess the level of difficulty when performing a life habit, the type of assistance required and the person's level of satisfaction in relation to the accomplishment of each life habit, to establish a profile of handicap situation in cancer patients and to assess the relationship between the level of accomplishment and the level of satisfaction. PATIENTS AND METHODS: The study sample consisted of 100 cancer patients. The Life Habits Assessment was used to measure the degree of accomplishment in 12 categories of life habits - daily living and social roles. RESULTS: Disruptions in the accomplishment of all life habit categories were revealed. The highest disruptions were observed in the following categories: education, recreation, and employment. The highest accomplishments were observed in communication, personal care, and fitness. Human assistance was required in the accomplishment of responsibility, community, employment and recreation. The highest person's level of satisfaction was observed in communication, personal care and interpersonal relations. The highest correlation between the level of accomplishment and the level of satisfaction were in mobility and residence, and the lowest correlation was in communication. CONCLUSION: The established profile of handicap situation in cancer patients should help specify rehabilitation objectives and may be a frame for individual treatment plan in accomplishing better social participation of cancer patients. PMID- 19810136 TI - Analysis of cutaneous melanoma in the province of Vojvodina. AB - PURPOSE: To analyse the incidence and mortality of melanoma in the province of Vojvodina - the north part of Serbia- from the epidemiologic and clinical point of view. PATIENTS AND METHODS: In this retrospective study, we used databases of the Cancer Registry of Vojvodina, Oncology Institute of Vojvodina in Sremska Kamenica; Clinical Centre of Vojvodina in Novi Sad; Institute of Public Health of Vojvodina in Novi Sad; and Institute of Public Health of Serbia in Belgrade. RESULTS: The incidence and mortality of melanoma in Vojvodina is permanently increasing over the last three decades. The number of newly diagnosed and deceased patients was higher in men and in women in Vojvodina than in central Serbia. Considering the incidence rate, the region of Vojvodina would be placed in the middle of the European countries. The results are more unfavorable with mortality, especially in male population, similar to the range of higher mortality rates among European countries. CONCLUSION: These upsetting facts call for improvement of primary and secondary prevention. Education of the population, continuous efforts to recognize groups of population at high risk for developing melanoma and detection of early phases of the disease increases the chances for cure and significantly reduces treatment costs. PMID- 19810137 TI - A virtual fluoroscopy system to verify seed positioning accuracy during prostate permanent seed implants. AB - PURPOSE: The purpose of this project was to develop a software platform to produce a virtual fluoroscopic image as an aid for permanent prostate seed implants. MATERIALS AND METHODS: Seed location information from a pre-plan was extracted and used as input to in-house developed software to produce a virtual fluoroscopic image. In order to account for differences in patient positioning on the day of treatment, the user was given the ability to make changes to the virtual image. RESULTS: The system has been shown to work as expected for all test cases. CONCLUSION: The system allows for quick (on average less than 10 sec) generation of a virtual fluoroscopic image of the planned seed pattern. The image can be used as a verification tool to aid the physician in evaluating how close the implant is to the planned distribution throughout the procedure and enable remedial action should a large deviation be observed. PMID- 19810138 TI - Dosimetric comparison of ring and ovoid applicators. AB - PURPOSE: To compare the dosimetry of different vaginal applicators used in the postoperative vaginal cuff irradiation. MATERIALS AND METHODS: In this model dosimetric study, standard ovoid applicator sets with 3 cap sizes (small/15 mm, medium/20 mm, and large/25 mm), and ring applicator sets with 3 diameter sizes (small/26 mm, medium/30 mm, large/34 mm) each with 3 different angles (30, 45, and 60 degrees) were used. Calculations were performed in Plato TPS (Treatment Planning System) as dwell positions of 2.5 mm step and equal dwell time for the sources. For ovoid applicators, the source positions 1-5 were loaded as active on each catheter, and in ring applicators the source positions 1-12, and 18-29; 1 14, and 19-32; 1-17, and 22-37 were loaded for small, medium and large ones, respectively. In addition to ICRU rectum reference point (R(1)), 4 different rectum reference points (RP(s)) (R(2)-R(5)) were defined 1 cm apart. The vaginal cuff RP (V(c)) was defined at 5 mm depth from the vaginal surface. The reference volume dose distribution was obtained according to the V(c) (100%) reference point. The width (w), max-min length (h, h'), and max-min thickness (t, t') of the reference isodoses were measured and max-min reference volume (v, v') was calculated. RESULTS: R(1) dose was higher for ovoids (43.1%) than ring (28.6%), in all applicators. The increase of R(1) dose was parallel to the increase of ovoid/ring size (70-89%). As the applicator (ovoid-ring) size increased, max thickness of the reference isodose also increased (19-50%). For the largest size, the max increase was 62% (74.1-119.88 cm(3)) and 93% (59.15-114.24 cm(3)) for v and v', respectively. However, the magnitude of the reference volume was independent of the ring angle. CONCLUSION: In the present study it was found that when ring applicators were used according to recommended dwell positions, max rectum reference dose decreased in comparison to ovoid applicators; on the other hand, it should be noted that the size of the reference volume also decreased significantly. PMID- 19810139 TI - Prognostic impact of HER2/neu protein in urothelial bladder cancer. Survival analysis of 80 cases and an overview of almost 20 years' research. AB - PURPOSE: This study was conducted to evaluate the quantitative assessment of HER2/neu immunohistochemical expression in urothelial bladder cancer in order to determine its prognostic significance. MATERIALS AND METHODS: Archival tumor tissue from 80 patients with primary urothelial carcinoma were analysed for HER2/neu immunohistochemical expression. A highly reproducible standardized procedure on a Bond-X automated slide stainer was used. RESULTS: HER2 protein was overexpressed in 41 of 80 patients (51.25%), demonstrating an increase in the expression rate corresponding to progressively advanced tumor stage (p=0.032) and tumor grade (p=0.0001). Kaplan-Meier analyses showed that positive membranous expression of HER2/neu was not associated with an increased probability of tumor recurrence (p=0.362). In contrast, HER2 scores correlated strongly with specific survival probability (p=0.002) and overall survival (p=0.025). Multivariate analysis revealed that only stage was an independent predictor of specific survival (p=0.016). HER2 expression was an independent predictor of specific survival with borderline statistical significance (p=0.08). CONCLUSION: HER2 overexpression represents a prognostic factor for adverse disease outcome. PMID- 19810140 TI - Type I collagen biomarkers in the diagnosis of bone metastases in breast cancer, lung cancer, urinary bladder cancer and prostate cancer. Comparison to CEA, CA 15 3, PSA and bone scintigraphy. AB - PURPOSE: In this study we evaluated the clinical usefulness of serum pro-I collagen peptide (PICP) and I collagen telopeptide (ICTP) as indicators of early bone metastases in patients with breast (BC), lung (LC), urinary bladder (UBC) and prostate cancer (PC). PATIENTS AND METHODS: 305 patients were examined. 145 had histologically confirmed BC (92 with bone metastases), 20 UBC (6 with bone metastases), 11 LC (3 with bone metastases) and 129 PC (68 with bone metastases). In BC patients we compared the PICP and ICTP levels with those of CA 15-3, CEA and bone scintigraphy. Patients with LC and UBC had PICP and ICTP measurements, PC patients had serum PICP, prostate specific antigen (PSA) measurements and bone scans. 104 healthy individuals served as controls. RESULTS: ICTP and CA 15-3 levels were significantly higher in patients with BC and bone metastases in comparison to patients without metastases (p <0.05), while PICP and CEA were only marginally higher. Significant correlation was observed between existence of bone metastases and ICTP levels (p <0.05). The sensitivity of PICP, ICTP, CEA and CA 15-3 was 28.1, 48.6, 42, and 78%, respectively and specificity was 83.9, 94, 65 and 86%, respectively. ICTP and CA 15-3 were the most reliable markers for early diagnosis of bone metastases in BC. PICP alone or with ICTP were not sensitive enough. Only CA 15-3 showed sensitivity 78% and specificity 86%. When combined CA 15-3, ICTP and CEA the sensitivity and specificity increased to 82% and 96%, respectively. Furthermore, PICP and PSA levels were significantly higher in patients with PC and bone metastases in comparison to patients with benign prostate hyperplasia (BPH) (p <0.0001) or in patients with PC without bone metastases (p <0.0005 for PICP and p <0.0001 for PSA). The co-evaluation of PICP and PSA improved the sensitivity (78%), specificity (96%), accuracy (97%) and positive predictive value (97%). In LC patients, ICTP levels differed significantly between patients with and without bone metastases (p=0.025). In UBC patients, PICP levels differed significantly between patients with and without bone metastases (p=0.017). CONCLUSION: ICTP and CA 15-3 are the most reliable markers for early diagnosis of bone metastases in BC patients. PICP could be useful for diagnosing early bone metastases of PC and combined with PSA and bone scan can be an additional tool in the follow-up of PC patients. For LC patients, ICTP showed a significant difference in the discrimination of patients with and without bone metastases. In UBC patients, PICP showed a significant difference in the discrimination of patients with and without bone metastases. PMID- 19810141 TI - Tumor size and karyometric variables in brain astrocytoma. AB - PURPOSE: To show any possible correlation of some karyometric variables with tumor size in patients with brain astrocytoma, in order to confirm karyometry as an objective histological method. PATIENTS AND METHODS: The study included 63 patients of different ages and both genders with brain astrocytoma histologically confirmed on the surgically removed material. In all patients maximal tumor excision was done, and all were postoperatively treated according to different therapeutic protocols. Tumor size (preoperative CT scan) was correlated with the duration of survival and the values of some karyometric tumor variables: area, density, maximal axis, mean axis, minimal axis, circumference, roundness, integrated optical density (IOD) and number of nuclei. RESULTS: Patients were separated into 3 groups according to the average tumor diameter. There were 34 cases of medium-sized tumors, 12 of small and 17 of large-sized tumors, and their respective survival was 83, 97 and 24 weeks. Patients with large tumors had statistically shorter survival compared to those with medium and small tumors (log-rank test, p=0.0122). Seven out of 9 examined karyometric variables were significantly related (p<0.05) to the tumor size: area, maximal axis, mean axis, minimal axis, circumference, roundness and IOD. CONCLUSION: Patients with larger tumors have shorter survival. The results of our morphometric analysis of the tumor cell nuclei, after correlation with CT findings, revealed that nuclear pleomorphism and larger nuclear size are associated with larger brain astrocytomas. PMID- 19810142 TI - Enhancement of taxane-induced cytotoxicity and apoptosis by gossypol in human breast cancer cell line MCF-7. AB - PURPOSE: Gossypol is a natural polyphenolic compound extracted from cotton plant (Gossypium species) which has shown potent inhibitory effect on cell growth of many types of cancers. In this study, we aimed to evaluate the interaction of gossypol with some conventional drugs known to be effective in the treatment of breast cancer, like taxanes, doxorubicin, gemcitabine, cisplatin and vinorelbine, in MCF-7 breast cancer cells. MATERIALS AND METHODS: The XTT viability assay was used to evaluate the cytotoxicity of various cytotoxic agents alone and in combination with gossypol in MCF-7 breast cancer cells. The combination effect analysis of Chou and Talalay was used to identify the most synergistic drug combinations. The possible synergistic effects of the combination of drugs on apoptosis were also evaluated by using two different apoptosis assays. RESULTS: We identified strong synergistic cytotoxic and apoptotic activity of gossypol with taxanes among all other studied cytotoxic drugs. CONCLUSION: This study provides proof that gossypol combined with taxanes may have potential as a novel future treatment for breast cancer. PMID- 19810143 TI - Expression of Fas/FasL in patients with oral lichen planus. AB - PURPOSE: To investigate the malignant potential of oral lichen planus (OLP) on the basis of expression of the Fas/FasL markers in healthy individuals (H), OLP patients and patients with squamous cell carcinoma (SCC). PATIENTS AND METHODS: 40 patients with OLP and two control groups were included in this research (H and patients with SCC). Immunohistochemistry for Fas and FasL was carried out using an avidin-biotin peroxidase complex method. RESULTS: Only a low percentage of infiltrating lymphocytes and no keratinocytes were Fas-positive in OLP specimens. The highest percentage of Fas-staining keratinocytes in our survey was identified mostly in H and patients with well-differentiated SCC. In most cases of SCC, OLP and H a high percentage of keratinocytes and lymphocytes were FasL-positive. FasL expression was negatively correlated with the degree of cell differentiation and apoptosis. Taking into consideration that all carcinomas in this survey were highly differentiated, it is not surprising that no statistically significant differences in FasL expression between H, OLP and SCC specimens were detected. CONCLUSION: Downregulation of Fas expression in keratinocytes and lymphocytes of OLP specimens, together with upregulation of FasL, may serve as initial prognostic biomarker in oral cancer development. PMID- 19810144 TI - Detection of human papillomavirus DNA by polymerase chain reaction and southern blot hybridization in colorectal cancer patients. AB - PURPOSE: The molecular mechanisms related to colorectal carcinogenesis are controversial. The purpose of this study was to evaluate the possible role of high-risk oncogenic human papillomavirus (HPV) types in the pathogenesis of colorectal cancer. PATIENTS AND METHODS: Tumor, and corresponding normal mucosal tissue specimens were obtained soon after surgery from 56 patients with colorectal adenocarcinoma. We studied both neoplastic and normal colon tissues for the presence of HPV types 6, 11, 16, 18, and 33. After the isolation of DNA, the presence of specific types of HPV DNA was determined by polymerase chain reaction (PCR) and southern blot hybridization. RESULTS: HPV DNA was detected in 46 (82.14 %) of 56 colorectal adenocarcinomas and in 18 (32 %) of 56 normal colonic mucosal tissue samples. Two or more HPV types were detected in 32 carcinoma samples. HPV type 18 (n= 40) and 33 (n= 32) were the most frequently detected types of HPVs in the tumor tissues. None of the normal mucosal specimens revealed HPV 18 DNA. The expression rate of HPV DNA in tumor tissue was significantly higher than that encountered in normal colonic mucosa (p <0.001). CONCLUSION: Detection of HPV DNA types 18 and 33 in most of the colorectal adenocarcinoma specimens suggests that HPVs may be related to carcinogenesis in glandular cells of the colorectal mucosa of our patient population. PMID- 19810145 TI - Lipid profile as a prognostic factor in cancer patients. AB - PURPOSE: The relationship between plasma lipid levels and neoplastic diseases is still unclear. The aim of this study was to analyse the lipid profile of individuals with non-Hodgkin's lymphoma (NHL) or prostate carcinoma (CaP) and to follow serum lipid levels changes in NHL patients according to their response to chemotherapy. PATIENTS AND METHODS: Forty-seven patients with NHL, 57 patients with CaP, two control groups composed of 29 and 43 age- and sex-matched healthy adults, related to NHL and CaP patients, respectively, were included in the study. Follow-up studies of NHL patients were carried out after the 3rd and 6th cycle of chemotherapy. RESULTS: Initial plasma cholesterol (Chol), HDL cholesterol (HDL-Chol) and phospholipids (PL) values were significantly lower in patients with NHL or CaP than in controls. Following chemotherapy, we noticed a progressive increase in lipid levels in NHL patients with complete remission (CR) and stable disease (SD), and further decrease in patients with the disease progression. CONCLUSION: Decreased plasma Chol, HDL-Chol and PL levels of patients with NHL or CaP can be considered as non-specific prognostic parameters in patients with these malignancies. PMID- 19810146 TI - Chemoembolization facilitates limb salvage surgery in stage III soft tissue sarcoma. AB - A 26 year-old male was referred to our unit because of a stage III soft tissue sarcoma in the shoulder girdle-axillary area and reduced forearm-distal arm strength. Imaging studies revealed that the tumor encompassed the axillary artery and brachial plexus. We chemoembolized it using vincristine, adriamycin and cyclophosphamide (VAC) plus gel foam and performed limb salvage surgery (LSS) afterwards. The patient received adjuvant chemotherapy (ifosfamide/mesna, adriamycin, and dacarbazine/MAID) and finally radiation therapy (RT; 6500 cGy total dose). Thirty-six months after the operation the patient remains free of disease, without local recurrence and excellent neurological recovery and functional rehabilitation. In stage III soft tissue sarcomas, especially in proximity with major nerve/arterial bundles, a multimodality approach is mandatory; chemoembolization is very effective in shrinking the tumor and defining its margins so as to make feasible a LSS. PMID- 19810147 TI - Synchronous renal cell carcinoma and multiple myeloma: report of two cases and review of the literature. AB - Coexistence of renal cell carcinoma (RCC) and multiple myeloma (MM) is an extremely rare condition. Appearance of synchronous RCC and MM was not reported independently so far. In this brief communication, we report 2 cases of synchronous RCC and MM, discuss common risk factors or pathogenetic mechanisms seen in either RCC or MM, point out the importance of IL-6 in this coexistence and provide some descriptive properties of all reported synchronous RCC and MM cases. PMID- 19810148 TI - Rapid improvement of extensive non-melanoma skin cancers with combination of 13 cis-retinoic acid and radiotherapy: report of three cases. AB - Extensive non-melanoma skin cancer (NMSC) constitutes a therapeutic challenge especially in old and debilitated patients. Applied treatments include surgical excision, MOHS micrographic surgery, cryosurgery, electrodesiccation, and radiotherapy. We present 3 elderly patients with extensive basal or squamous cell carcinomas and poor general condition who were treated with a combination of 13 cis-retinoic acid 1 mg/kg daily and radiotherapy 2.5 Gy daily. The treatment resulted in rapid improvement of the tumors with significant reduction of their size. PMID- 19810149 TI - Erythropoietin and radiotherapy in lung cancer patients. AB - PURPOSE: To evaluate the effectiveness of recombinant human erythropoietin (rhEPO) in relation to low hemoglobin (Hb) level, overall tumor response rates, and rhEPO adverse events in patients with lung cancer undergoing radiotherapy (RT). PATIENTS AND METHODS: Thirteen consecutive patients were included. All of them had measurable tumor before RT. 150 IU/kg of rhEPO-alpha or -beta were administered 3 times per week, 7-10 days before RT. The target Hb value was 13 g/dl. Tumor response was assessed 6 weeks after completion of RT. RESULTS: Response to rhEPO was seen 62% (n=8) of the patients. Weekly mean Hb increment was 0.69 g/dl (range 0.42-1). The mean Hb value during RT was 13.2 g/dl (range 9 14.7) in responding patients, and 10.7 g/dl (range 9.7-11.8) in non-responding patients (p=0.005). Overall response rates to RT were significantly higher in responding than in non-responding patients (p=0.034). CONCLUSION: rhEPO increased Hb levels in lung cancer patients undergoing RT. However, safety, and more importantly, indications need further clarifications. PMID- 19810150 TI - Massive osteoradionecrosis of facial bones and soft tissues. AB - Osteoradionecrosis (ORN) is one of the most serious and uncommon complications in head and neck irradiation for cancer. It is defined as a combination of necrotic soft tissue and bone not being able to heal spontaneously, it demonstrates a general resistance to antibiotics and requires conservative surgical management. Even with modern radiation therapy, its incidence is highly unpredictable and varies between 4-30%. We report on a patient with a huge open cavitation in the cheek, communicating with the mouth and extending to contralateral periodontal gingival and temporal fossa. He had been treated with radiation therapy for nasopharyngeal cancer 5 years ago and presented with restriction of the opening of the mouth. Osteonecrosis complicated with osteomyelitis was evident in bilateral mandible and maxillary bones and the temporal bone. The ramus of the mandible and zygomatic arc were resected, subtotal maxillectomy was performed and the defect was repaired by a free double island flap from the scapular and parascapular osteocutaneous latissimus dorsi muscle flap supplied by subscapular artery. To our knowledge, this is the most extensive bone and soft tissue destruction due to radiation reported in the literature. PMID- 19810151 TI - Alveolar rhabdomyosarcoma mimicking acute leukemia at presentation. PMID- 19810152 TI - Glomus cells tumors in the male genital area. PMID- 19810153 TI - Multiple second primary tumors in a woman following breast cancer. PMID- 19810154 TI - Governmental strategies for cancer control, prevention, care and management in Greece. AB - The Greek government through the Ministry of Health and the World Health Organisation (WHO) are concerned about the increase in cancer cases and are implementing actions related to its control. The WHO produced a guide which is a response to the World Health Assembly Resolution on Cancer Prevention and Control, adopted in May 2005. This WHO guide calls on member states to intensify action against cancer by developing and reinforcing cancer control programs. The Greek Ministry of Health has established a strategy unit and relevant health policies. The need to act comes from the fact that 11 million new cancer cases are diagnosed yearly, worldwide. The most common deaths globally are from lung cancer (1.3 million), from gastric cancer (1 million), liver cancer (662.000), colorectal cancer (665.000) and breast cancer (502.000), yearly. Some cancers in Greece are more common. There are certain risk factors on the basis of which one can take preventative measures related to the numerous chemical, biological and natural factors in the environment. Recently, the Greek Ministry of Health organized a national plan of action against cancer. This starts with monitoring scientific societies and social groups and organizations in order to promote prevention as well as an improvement in the quality of cancer management. The targets are the collection of reliable information, primary and secondary prevention, effective treatment and a better quality of life for the patient. PMID- 19810155 TI - Cancer and Roman medicine. AB - In this article we present the opinions of some eminent Roman physicians about cancer. We emphasize the points of view of Galen, famous Roman physician of Greek origin whose doctrines dominated medicine for more than 15 centuries. We then cite the opinions about cancer of some other outstanding physicians of that period like Celsus, Pliny, Leonidis of Alexandria, Aretaeus of Cappadocia, Soranus of Ephesus etc. We also quote some examples of cancer and we describe what those physicians thought of its pathogenesis, diagnosis and treatment. PMID- 19810156 TI - Primary access to vehicles increases risky teen driving behaviors and crashes: national perspective. AB - OBJECTIVE: The goal was to explore teen driver vehicle access and its association with risky driving behaviors and crashes. METHODS: A nationally representative, school-based survey of 2167 ninth-, 10th-, and 11th-graders examined patterns of vehicle access (primary access [ie, the teen is the main driver of the vehicle] versus shared access) and associated driving exposure, risky driving behaviors, and sociodemographic factors. RESULTS: Seventy percent of drivers reported having primary access to vehicles. They were more likely to be white, to be in 11th grade, to attend schools with higher socioeconomic levels, to have mostly A/B grades, to have a job, to drive a pickup truck, and to drive more hours per week but were not more or less likely to consume alcohol or to wear seat belts while driving. Compared with drivers with shared access, drivers with primary access reported more than twice the crash risk (risk ratio [RR]: 2.05 [95% confidence interval [CI]: 1.41-2.99]) and higher likelihoods of using cellular telephones while driving (RR: 1.23 [95% CI: 1.12-1.35]) and speeding> or =10 mph above the posted limit (RR: 1.24 [95% CI: 1.11-1.40]). CONCLUSIONS: Primary access of novice teen drivers to vehicles is highly prevalent in the United States. This practice is a dangerous norm, because primary access is associated with risky driving behaviors. Healthcare providers and schools should consider counseling parents to discourage giving novice teen drivers primary access to vehicles. In communities where teens require primary access (eg, due to limited public transportation options), greater efforts should be made to promote safe behaviors. PMID- 19810157 TI - US reimbursement systems encourage fraud and overutilisation. PMID- 19810158 TI - Cuts in reimbursement proposed for radiation oncology in the US. PMID- 19810159 TI - International Liver Cancer Association Annual Conference. PMID- 19810160 TI - [Anticancer therapeutics: should a different therapeutic concept be considered?]. PMID- 19810161 TI - [Homage to Olivier Hartmann (1944-2009)]. PMID- 19810162 TI - ["Anapath", prescription and prediction guide in cancerology]. PMID- 19810163 TI - [The inhibition of the Sonic Hedgehog (SHh) pathway improves the effects of chemotherapy in a model of ductal carcinoma of the pancreas in mice]. PMID- 19810164 TI - [Whole body robotic stereotaxic radiotherapy with Cyberknife: introduction ]. PMID- 19810165 TI - Introduction to Brazilian Guidelines to Diagnosis, Treatment,and Monitoring for Gaucher Disease, Fabry Disease, Mucopolysaccharidosis I, and Pompe Disease. PMID- 19810166 TI - Why ceftazidime? PMID- 19810167 TI - The in-vitro activity of ceftazidime against clinically important pathogens. AB - A total of 1414 strains of Gram-negative bacilli and 250 strains of Gram-positive cocci were tested for antimicrobial susceptibility using the broth dilution method. The strains were clinical isolates from 26 hospitals in the Rhein-Main area. All the strains were tested against ceftazidime and ten other cephalosporin antibiotics. Ceftazidime was found to exhibit an in-vitro activity against Enterobacteriaceae similar to that of cefotaxime, moxalactam and other new members of the cephalosporin class, but was more active against Acinetobacter species. Ceftazidime shows extremely high activity both against carbenicillin sensitive and -resistant strains of Pseudomonas aeruginosa. Similarly, ampicillin sensitive and -resistant strains of Haemophilus influenzae are susceptible to ceftazidime. The activity of ceftazidime against Staphylococcus aureus is limited and, like other cephalosporins, it is inactive against Streptococcus faecalis. PMID- 19810168 TI - A comparison of the in-vitro properties of ceftazidime with those of new broad spectrum cephalosporins and gentamicin. PMID- 19810169 TI - In-vitro activity of ceftazidime compared with other beta-lactam antibiotics. AB - Ceftazidime and ceftriaxone (Ro 13-9904), two new parenteral cephalosporins, were compared with other beta-lactam antibiotics against a total of 237 recent clinical isolates. Both ceftazidime and ceftriaxone had good activity against the Enterobacteriaceae, with 90% of isolates being inhibited by 0.5 mg/l of ceftazidime and by 0.25 mg/l of ceftriaxone. Against Pseudomonas aeruginosa, ceftazidime was about four times more active than ceftriaxone which, in turn, was at least eight times more active than carbenicillin. Both ceftazidime and ceftriaxone were highly active against beta-lactamase producing and non-beta lactamase producing Haemophilus influenzae. Against the Bacteroides fragilis group, neither ceftazidime nor ceftriaxone showed good activity, the MIC90 for ceftazidime being > 128 mg/l and the MIC90 for ceftriaxone being 32 mg/l. Both ceftazidime and ceftriaxone were less active than cephalothin against Staphylococcus aureus. PMID- 19810170 TI - In-vitro studies with ceftazidime against aerobic gram-negative bacilli and Bacteroides fragilis group. AB - The in-vitro susceptibility of recent clinical isolates of Gram-negative bacilli has been assessed for ceftazidime and compared to amikacin, gentamicin, carbenicillin, piperacillin, cefoperazone, moxalactam, ceftriaxone and ceftizoxime. Using the ICS-WHO agar dilution method, we found that ceftazidime was the most active beta-lactam agent tested against 147 isolates of Pseudomonas aeruginosa, with a mode MIC=2 mg/l and all but 2% of isolates inhibited at 32 mg/l. 100% of indole-positive and negative Proteus spp., 90% of Citrobacter spp., 100% of Acinetobacter spp. and 98% of Enterobacter spp. were inhibited. A total of 142 isolates from the latter 5 groups of organisms were tested. Cefoperazone and moxalactam were slightly more active by weight than ceftazidime versus Enterobacter spp., but against other Gram-negative bacilli ceftazidime was similar or more potent. Additionally, 72 clinical isolates of Bacteroides fragilis group were tested against ceftazidime, cefoperazone, ceftizoxime, ceftriaxone, and cefoxitin. Against these organisms cefoxitin and ceftizoxime were most active. Ceftazidime demonstrates potent in-vitro activity against Ps. aeruginosa and Enterobacteriaceae, but it is relatively less active against Bacteroides fragilis group than cefoxitin and ceftizoxime. PMID- 19810171 TI - An in-vitro comparison of new cephalosporins with special reference to Pseudomonas aeruginosa. AB - MICs of cefoperazone, cefotaxime, ceftazidime and moxalactam were determined for staphylococci, Enterobacteriaceae, Haemophilus influenza and Neisseria gonorrhoeae. The new compounds are 10 to 100 times more active in vitro against Enterobacteriaceae and H. influenzae (including penicillinase-producing strains) than the older cephalosporins. Against N. gonorrhoeae, including penicillinase producing strains, cefotaxime appeared to be the most active compound (MIC as low as 0.002 mg/l), while the other new cephalosporins were approximately as active as cefuroxime. Cefoperazone, cefotaxime, cefsulodin, ceftazidime, ceftriaxone and moxalactam were tested against carbenicillin susceptible (Cbs) and carbenicillin resistant (Cb(R)) strains of Pseudomonas aeruginosa. Against Cb(S) strains the lowest MIC values were obtained with ceftazidime (1 mg/l) followed by cefsulodin (1-2 mg/l), cefoperazone (2-4 mg/l), ceftriaxone (4 mg/l), moxalactam (4-8 mg/l) and cefotaxime (8-16 mg/l). Usually the susceptibility to cephalosporins of Cb(R) strains was found decreased for all compounds, but the least for ceftazidime. Considerable cross resistance with carbenicillin was noted for cefsulodin and cefoperazone only. Lysates of 19 CbR strains have been screened for the presence of constitutive beta-lactamases. Three enzyme types were found: pI 5.4 (n=8) pl 5.3 (n=5) and pI 5.7 (n=6). PMID- 19810172 TI - An evaluation of the activity of cephalosporins against Pseudomonas aeruginosa. AB - We investigated the in-vitro activity of 14 antibiotics against 200 distinct isolates of Pseudomonas aeruginosa obtained from intensive care patients. Ceftazidime was the most active beta-lactam and 16 mg/l inhibited 88% of the strains. Of 55 two-drug combinations, ceftazidime plus tobramycin was the most effective and 98.5% of strains were sensitive. Synergism between these two drugs was demonstrated for 12 of the 23 strains tested. Ceftazidime-inactivating enzymes may be induced in Ps. aeruginosa by cefoxitin. PMID- 19810173 TI - Comparative in-vitro activity of cefsulodin and ceftazidime against ticarcillin resistant Pseudomonas aeruginosa. AB - The in-vitro antibacterial activity of ceftazidime was compared with that of cefsulodin and ticarcillin against 140 strains of Pseudomonas aeruginosa. Against 123 clinical isolates, geometric mean MICs were 1499mg/l for ticarcillin, 15 mg/l for cefsulodin and 2.1 mg/l for ceftazidime. Against constitutive beta-lactamase producers (93 strains), and according to the type of enzyme (TEM-1, carbenicillinases, oxacillinases), ceftazidime had similar activity but MICs of cefsulodin were particularly affected by highly ticarcillin-resistant (carbenicillinase pIs 53, 5.7 and 575) strains. Against ticarcillin-resistant (constitutive beta-lactamase non-producers) and ticarcillin-susceptible strains, the MICs of ceftazidime were slightly lower than those of cefsulodin. PMID- 19810174 TI - Comparison of the in-vitro activities of ceftazidime and new cephalosporins against 107 strains of Pseudomonas aeruginosa and 249 strains of cefazolin resistant Enterobacteriaceae. AB - Three hundred and fifty-six bacterial strains, including 107 strains of Pseudomonas aeruginosa, were tested with ceftazidime and five other agents. All bacterial strains were resistant to cefazolin but were sensitive to 32 mg/l of ceftazidime or less. PMID- 19810175 TI - In-vitro activity of ceftazidime, a beta-lactamase stable cephalosporin. AB - The in-vitro activity and beta-lactamase stability of ceftazidime were evaluated against 700 Gram-positive and Gram-negative bacteria. Ceftazidime was less active than penicillins or older cephalosporins against Staphylococcus spp. and Streptococcus spp., and it did not inhibit Streptococcusfaecalis. However, ceftazidime was as active as moxalactam and cefoperazone against Escherichia coli, Klebsiella and Proteus mirabilis with MICs less than 0.2 mg/l. Ceftazidime also inhibited Enterobacter, Citrobacter, Salmonella and Shigella at concentrations below 0.2 mg/l. Most Morganella, Pr. rettgeri, Pr. vulgaris and Pr. inconstans were inhibited at concentrations below 1 mg/l similar to concentrations for moxalactam, cefoperazone, and cefotaxime. Ceftazidime was the most active agent tested against Ps. aeruginosa, with a mean MIC of 1.6 mg/l. It inhibited carbenicillin, piperacillin, cefoperazone, cefsulodin resistant Pseudomonas. PMID- 19810176 TI - Behaviour of ceftazidime towards beta-lactamases. AB - from Escherichia coli, Enterobacter cloacae, Proteus morganii and Pseudomonas aeruginosa, but the geometric mean of the inhibition constant Ki used to measure this interaction was 120 and almost 700 times higher with ceftazidime than with cefotaxime and cefuroxime, respectively. Two other beta-lactamases isolated from Pr. vulgaris and Klebsiella oxytoca also showed little or no interaction with ceftazidime. A complementary microbiological technique, the "double-disc" technique, indicated that ceftazidime was more stable than cefotaxime to all beta lactamases tested. Moxalactam, however, appeared to have slightly greater stability. PMID- 19810177 TI - [Pleural empyema. Incidence, puncture therapy, closed pleural cavity drainage. Part 1]. PMID- 19810178 TI - [Pathomorphism of tuberculous pleurisy in elderly and senile individuals]. AB - The pathomorphism of tuberculous exudative pleurisy in elderly and senile individuals has been investigated in the past 30 years. Comparison of clinical, X ray, laboratory, morphological, and immunological data in patients of the first (1978-1987) and second (1998-2007) periods has revealed the signs characteristic of the negative pathomorphism of the disease: progressive latency of tuberculous pleurisy and a lower Mantoux reaction, a lower detection rate for epithelioid giant-cell granulomas in the patients' biopsy specimens, a higher rate of pleural tuberculous inflammation, an increased frequency of comorbidity, a higher drug resistance rate in Mycobacterium tuberculosis, and decreased therapeutic efficiency. In this connection, elderly and senile persons with pleural effusion should be examined in a tuberculosis dispensary, with the bacteriological and morphological verification of the diagnosis being made. Their treatment should comprise the currently available antituberculous and pathogenetic agents. PMID- 19810179 TI - [Diagnostic value of interferon-gamma in the serum of patients with pulmonary tuberculosis and lung cancer diseases]. PMID- 19810180 TI - [Drug resistance in Mycobacterium tuberculosis during chronic intoxication by tobacco: clinical and experimental studies]. AB - Case histories were studied in 405 smoking patients (including 84 inveterate tobacco smokers) (a study group) and 193 nonsmoking patients (a control group) with new-onset pulmonary tuberculosis. Smoking was found to lead to a significant increase in the cases of drug resistance (including multidrug resistance (MDR)) in Mycobacterium tuberculosis (MBT): the resistant strains were 42.9% in the smokers and 54.4% in the inveterate tobacco smokers versus 31.9% in the control group (p < 0.01). The experiments showed that after two MBT strains (H37 Ra and 4688) had grown on the tobacco smoke condensate media, 57.1% of the experimental cultures were resistant to both one and several agents (including MDR), as shown by a study using the absolute concentration method. PMID- 19810181 TI - [Immunomorphological characteristics of experimental granulomatous inflammation caused by Mycobacterium avium]. AB - The results of the topical arrangement of T and B lymphocytes in the lung tissue granulomatous masses of the mice susceptible (C57BL/6Ycit) and resistant (I/StSnEgYCit) to aerosol inoculation with M. avium 724R in a dose of 2 x 10(3) CFU/mice are presented by means of labeled monoclonal antibodies. At week 8 of the inoculation, CD4+ lymphocytes were located both among the granuloma cells in the diffuse fashion and as small clusters and in the interalveolar and interlobular septa in the B6 mice and these were mainly in the granuloma in the I/St mice. At week 16, CD8+ lymphocytes were found in the peripheral granuloma portions in the B6 mice and these were diffusely throughout the area of granulomatous masses in the I/St mice. In the B6 mice, CD19+ lymphocytes formed cell aggregates from week 8 of the inoculation; and in the I/st mice, these were diffusely located among the granuloma cells and abundantly in the interalveolar and interlohular septa. PMID- 19810182 TI - [Complex evaluation of work conditions and health state of oil industry workers]. AB - The authors studied influence of occupational environment and work process factors on health of workers engaged into oil extraction. Analysis covered 7487 oil industry workers. Locomotory system and peripheral nervous system diseases, arterial hypertension, ENT diseases, gastro-intestinal disorders appeared to play leading role in the morbidity structure revealed. The studies helped to specify measures to preserve the oil workers health. PMID- 19810183 TI - [Occupational health in factory workers engaged into iron quartzite enrichment, prophylactic measures]. AB - Outstanding feature of work conditions on the processing plants studied is infrasound present at some workplaces. Relative risk parameters according to case and day shares prove high and nearly complete relationships between the work conditions and ear and mastoid diseases among the processing plant workers. The authors set priority nosologic entities to form general level of transitory disablement morbidity. PMID- 19810184 TI - [Cardio-vascular diseases in individuals having long-term effects of acute sarin and soman poisonings]. AB - Adding complex clinical assessment of cardio-vascular functional capacity to diagnostic workup program for individuals having long-term effects of acute sarin and soman poisonings enables simultaneous characteristic of various hemodynamic changes and their relationships, early diagnosis of hemodynamic disorders, opportune diagnosis of clinical cardiovascular syndromes. PMID- 19810185 TI - Associations between parenting styles and teen driving, safety-related behaviors and attitudes. AB - OBJECTIVE: The goal was to explore the association between parenting style and driving behaviors. METHODS: The 2006 National Young Driver Survey gathered data on driving safety behaviors from a nationally representative sample of 5665 ninth , 10th-, and 11th-graders. A parenting style variable was based on adolescent reports and separated parents into 4 groups, (1) authoritative (high support and high rules/monitoring), (2) authoritarian (low support and high rules/monitoring), (3) permissive (high support and low rules/monitoring), and (4) uninvolved (low support and low rules/monitoring). Associations between parenting style and driving behaviors and attitudes were assessed. RESULTS: One half of parents were described as authoritative, 23% as permissive, 8% as authoritarian, and 19% as uninvolved. Compared with teens with uninvolved parents, those with authoritative parents reported one half the crash risk in the past year (odds ratio [OR]: 0.47 [95% confidence interval [CI]: 0.26-0.87]), were 71% less likely to drive when intoxicated (OR: 0.29 [95% CI: 0.19-0.44]), and were less likely to use a cellular telephone while driving (OR: 0.71 [95% CI: 0.50-0.99]). Teens with authoritative or authoritarian parents reported using seat belts nearly twice as often (authoritative: OR: 1.94 [95% CI: 1.49 -2.54]; authoritarian: OR: 1.85 [95% CI: 1.08 -3.18]) and speeding one half as often (authoritative: OR: 0.47 [95% CI: 0.36-0.61]; authoritarian: OR: 0.63 [95% CI: 0.40-0.99]) as teens with uninvolved parents. No significant differences in crash risk or seat belt use were found between permissive and uninvolved parents. CONCLUSIONS: Clinicians should encourage parents to set rules and to monitor teens' driving behaviors, in a supportive context. PMID- 19810186 TI - Howell-Jolly body-like inclusions in neutrophils. PMID- 19810187 TI - Abstracts of the Annual Meeting of the Swiss Society of Gastroenterology, Swiss Society for Visceral Surgery, and the Swiss Association for the Study of the Liver. September 17-18, 2009. Zurich, Switzerland. PMID- 19810188 TI - Surrogate mobility and orientation affect the early neurobehavioral development of infant rhesus macaques (Macaca mulatta). AB - A biological mother's movement appears necessary for optimal development in infant monkeys. However, nursery-reared monkeys are typically provided with inanimate surrogate mothers that move very little. The purpose of this study was to evaluate the effects of a novel, highly mobile surrogate mother on motor development, exploration, and reactions to novelty. Six infant rhesus macaques (Macaca mulatta) were reared on mobile hanging surrogates (MS) and compared to six infants reared on standard stationary rocking surrogates (RS) and to 9-15 infants reared with their biological mothers (MR) for early developmental outcome. We predicted that MS infants would develop more similarly to MR infants than RS infants. In neonatal assessments conducted at Day 30, both MS and MR infants showed more highly developed motor activity than RS infants on measures of grasping (p = .009), coordination (p = .038), spontaneous crawl (p = .009), and balance (p = .003). At 2-3 months of age, both MS and MR infants displayed higher levels of exploration in the home cage than RS infants (p = .016). In a novel situation in which only MS and RS infants were tested, MS infants spent less time near their surrogates in the first five minutes of the test session than RS infants (p = .05), indicating a higher level of comfort. Collectively, these results suggest that when nursery-rearing of infant monkeys is necessary, a mobile hanging surrogate may encourage more normative development of gross motor skills and exploratory behavior and may serve as a useful alternative to stationary or rocking surrogates. PMID- 19810189 TI - Proceedings of the XXXVI International Congress of Physiological Sciences. July 27- August 1, 2009. Kyoto, Japan. PMID- 19810190 TI - Ardipithecus ramidus and the paleobiology of early hominids. AB - Hominid fossils predating the emergence of Australopithecus have been sparse and fragmentary. The evolution of our lineage after the last common ancestor we shared with chimpanzees has therefore remained unclear. Ardipithecus ramidus, recovered in ecologically and temporally resolved contexts in Ethiopia's Afar Rift, now illuminates earlier hominid paleobiology and aspects of extant African ape evolution. More than 110 specimens recovered from 4.4-million-year-old sediments include a partial skeleton with much of the skull, hands, feet, limbs, and pelvis. This hominid combined arboreal palmigrade clambering and careful climbing with a form of terrestrial bipedality more primitive than that of Australopithecus. Ar. ramidus had a reduced canine/premolar complex and a little derived cranial morphology and consumed a predominantly C3 plant-based diet (plants using the C3 photosynthetic pathway). Its ecological habitat appears to have been largely woodland-focused. Ar. ramidus lacks any characters typical of suspension, vertical climbing, or knuckle-walking. Ar. ramidus indicates that despite the genetic similarities of living humans and chimpanzees, the ancestor we last shared probably differed substantially from any extant African ape. Hominids and extant African apes have each become highly specialized through very different evolutionary pathways. This evidence also illuminates the origins of orthogrady, bipedality, ecology, diet, and social behavior in earliest Hominidae and helps to define the basal hominid adaptation, thereby accentuating the derived nature of Australopithecus. PMID- 19810191 TI - The geological, isotopic, botanical, invertebrate, and lower vertebrate surroundings of Ardipithecus ramidus. AB - Sediments containing Ardipithecus ramidus were deposited 4.4 million years ago on an alluvial floodplain in Ethiopia's western Afar rift. The Lower Aramis Member hominid-bearing unit, now exposed across a > 9-kilometer structural arc, is sandwiched between two volcanic tuffs that have nearly identical 40Ar/39Ar ages. Geological data presented here, along with floral, invertebrate, and vertebrate paleontological and taphonomic evidence associated with the hominids, suggest that they occupied a wooded biotope over the western three-fourths of the paleotransect. Phytoliths and oxygen and carbon stable isotopes of pedogenic carbonates provide evidence of humid cool woodlands with a grassy substrate. PMID- 19810192 TI - Taphonomic, avian, and small-vertebrate indicators of Ardipithecus ramidus habitat. AB - Thousands of vertebrate specimens were systematically collected from the stratigraphic interval containing Ardipithecus ramidus. The carcasses of larger mammals were heavily ravaged by carnivores. Nearly 10,000 small-mammal remains appear to be derived primarily from decomposed owl pellets. The rich avifauna includes at least 29 species, mostly nonaquatic forms. Modern analogs of the most abundant birds and of a variety of rodents are associated with mesic woodland environments distant from large water bodies. These findings support inferences from associated geological, isotopic, invertebrate, and large-vertebrate assemblages. The combined results suggest that Ar. ramidus occupied a wooded Pliocene habitat. PMID- 19810193 TI - Macrovertebrate paleontology and the Pliocene habitat of Ardipithecus ramidus. AB - A diverse assemblage of large mammals is spatially and stratigraphically associated with Ardipithecus ramidus at Aramis. The most common species are tragelaphine antelope and colobine monkeys. Analyses of their postcranial remains situate them in a closed habitat. Assessment of dental mesowear, microwear, and stable isotopes from these and a wider range of abundant associated larger mammals indicates that the local habitat at Aramis was predominantly woodland. The Ar. ramidus enamel isotope values indicate a minimal C4 vegetation component in its diet (plants using the C4 photosynthetic pathway), which is consistent with predominantly forest/woodland feeding. Although the Early Pliocene Afar included a range of environments, and the local environment at Aramis and its vicinity ranged from forests to wooded grasslands, the integration of available physical and biological evidence establishes Ar. ramidus as a denizen of the closed habitats along this continuum. PMID- 19810194 TI - The Ardipithecus ramidus skull and its implications for hominid origins. AB - The highly fragmented and distorted skull of the adult skeleton ARA-VP-6/500 includes most of the dentition and preserves substantial parts of the face, vault, and base. Anatomical comparisons and micro-computed tomography-based analysis of this and other remains reveal pre-Australopithecus hominid craniofacial morphology and structure. The Ardipithecus ramidus skull exhibits a small endocranial capacity (300 to 350 cubic centimeters), small cranial size relative to body size, considerable midfacial projection, and a lack of modern African ape-like extreme lower facial prognathism. Its short posterior cranial base differs from that of both Pan troglodytes and P. paniscus. Ar. ramidus lacks the broad, anteriorly situated zygomaxillary facial skeleton developed in later Australopithecus. This combination of features is apparently shared by Sahelanthropus, showing that the Mio-Pliocene hominid cranium differed substantially from those of both extant apes and Australopithecus. PMID- 19810195 TI - Paleobiological implications of the Ardipithecus ramidus dentition. AB - The Middle Awash Ardipithecus ramidus sample comprises over 145 teeth, including associated maxillary and mandibular sets. These help reveal the earliest stages of human evolution. Ar. ramidus lacks the postcanine megadontia of Australopithecus. Its molars have thinner enamel and are functionally less durable than those of Australopithecus but lack the derived Pan pattern of thin occlusal enamel associated with ripe-fruit frugivory. The Ar. ramidus dental morphology and wear pattern are consistent with a partially terrestrial, omnivorous/frugivorous niche. Analyses show that the ARA-VP-6/500 skeleton is female and that Ar. ramidus was nearly monomorphic in canine size and shape. The canine/lower third premolar complex indicates a reduction of canine size and honing capacity early in hominid evolution, possibly driven by selection targeted on the male upper canine. PMID- 19810196 TI - Careful climbing in the Miocene: the forelimbs of Ardipithecus ramidus and humans are primitive. AB - The Ardipithecus ramidus hand and wrist exhibit none of the derived mechanisms that restrict motion in extant great apes and are reminiscent of those of Miocene apes, such as Proconsul. The capitate head is more palmar than in all other known hominoids, permitting extreme midcarpal dorsiflexion. Ar. ramidus and all later hominids lack the carpometacarpal articular and ligamentous specializations of extant apes. Manual proportions are unlike those of any extant ape. Metacarpals 2 through 5 are relatively short, lacking any morphological traits associable with knuckle-walking. Humeral and ulnar characters are primitive and like those of later hominids. The Ar. ramidus forelimb complex implies palmigrady during bridging and careful climbing and exhibits none of the adaptations to vertical climbing, forelimb suspension, and knuckle-walking that are seen in extant African apes. PMID- 19810197 TI - The pelvis and femur of Ardipithecus ramidus: the emergence of upright walking. AB - The femur and pelvis of Ardipithecus ramidus have characters indicative of both upright bipedal walking and movement in trees. Consequently, bipedality in Ar. ramidus was more primitive than in later Australopithecus. Compared with monkeys and Early Miocene apes such as Proconsul, the ilium in Ar. ramidus is mediolaterally expanded, and its sacroiliac joint is located more posteriorly. These changes are shared with some Middle and Late Miocene apes as well as with African apes and later hominids. However, in contrast to extant apes, bipedality in Ar. ramidus was facilitated by craniocaudal shortening of the ilium and enhanced lordotic recurvature of the lower spine. Given the predominant absence of derived traits in other skeletal regions of Ar. ramidus, including the forelimb, these adaptations were probably acquired shortly after divergence from our last common ancestor with chimpanzees. They therefore bear little or no functional relationship to the highly derived suspension, vertical climbing, knuckle-walking, and facultative bipedality of extant African apes. PMID- 19810198 TI - Combining prehension and propulsion: the foot of Ardipithecus ramidus. AB - Several elements of the Ardipithecus ramidus foot are preserved, primarily in the ARA-VP-6/500 partial skeleton. The foot has a widely abducent hallux, which was not propulsive during terrestrial bipedality. However, it lacks the highly derived tarsometatarsal laxity and inversion in extant African apes that provide maximum conformity to substrates during vertical climbing. Instead, it exhibits primitive characters that maintain plantar rigidity from foot-flat through toe off, reminiscent of some Miocene apes and Old World monkeys. Moreover, the action of the fibularis longus muscle was more like its homolog in Old World monkeys than in African apes. Phalangeal lengths were most similar to those of Gorilla. The Ardipithecus gait pattern would thus have been unique among known primates. The last common ancestor of hominids and chimpanzees was therefore a careful climber that retained adaptations to above-branch plantigrady. PMID- 19810199 TI - The great divides: Ardipithecus ramidus reveals the postcrania of our last common ancestors with African apes. AB - Genomic comparisons have established the chimpanzee and bonobo as our closest living relatives. However, the intricacies of gene regulation and expression caution against the use of these extant apes in deducing the anatomical structure of the last common ancestor that we shared with them. Evidence for this structure must therefore be sought from the fossil record. Until now, that record has provided few relevant data because available fossils were too recent or too incomplete. Evidence from Ardipithecus ramidus now suggests that the last common ancestor lacked the hand, foot, pelvic, vertebral, and limb structures and proportions specialized for suspension, vertical climbing, and knuckle-walking among extant African apes. If this hypothesis is correct, each extant African ape genus must have independently acquired these specializations from more generalized ancestors who still practiced careful arboreal climbing and bridging. African apes and hominids acquired advanced orthogrady in parallel. Hominoid spinal invagination is an embryogenetic mechanism that reoriented the shoulder girdle more laterally. It was unaccompanied by substantial lumbar spine abbreviation, an adaptation restricted to vertical climbing and/or suspension. The specialized locomotor anatomies and behaviors of chimpanzees and gorillas therefore constitute poor models for the origin and evolution of human bipedality. PMID- 19810200 TI - Reexamining human origins in light of Ardipithecus ramidus. AB - Referential models based on extant African apes have dominated reconstructions of early human evolution since Darwin's time. These models visualize fundamental human behaviors as intensifications of behaviors observed in living chimpanzees and/or gorillas (for instance, upright feeding, male dominance displays, tool use, culture, hunting, and warfare). Ardipithecus essentially falsifies such models, because extant apes are highly derived relative to our last common ancestors. Moreover, uniquely derived hominid characters, especially those of locomotion and canine reduction, appear to have emerged shortly after the hominid/chimpanzee divergence. Hence, Ardipithecus provides a new window through which to view our clade's earliest evolution and its ecological context. Early hominids and extant apes are remarkably divergent in many cardinal characters. We can no longer rely on homologies with African apes for accounts of our origins and must turn instead to general evolutionary theory. A proposed adaptive suite for the emergence of Ardipithecus from the last common ancestor that we shared with chimpanzees accounts for these principal ape/human differences, as well as the marked demographic success and cognitive efflorescence of later Plio Pleistocene hominids. PMID- 19810201 TI - Primary progressive aphasia and caregiver issues. PMID- 19810202 TI - Germany to host World Health Summit. PMID- 19810203 TI - Diabetes and the public's health. PMID- 19810204 TI - [3rd communication regarding changes in the guideline for public health determination of organic coatings in contact with drinking water (organic coating guideline)]. PMID- 19810207 TI - Cardiac care is better in the hands of a cardiologist but there are still regional differences within Europe. PMID- 19810208 TI - From mechanics to genetics in cardiology. PMID- 19810209 TI - Clinical conundrum. Intraocular mass. PMID- 19810210 TI - How to... Perform a cystotomy. PMID- 19810212 TI - The companion interview: Noel Fitzpatrick. PMID- 19810213 TI - Flash, bang, what? PMID- 19810214 TI - Differential effects of non-steroidal anti-inflammatory drugs on mitochondrial dysfunction during oxidative stress. AB - We investigated the effects of several non-steroidal anti-inflammatory drugs on swelling related properties of mitochondria, with an emphasis on compounds that are marketed and utilized topically in the eye (nepafenac, ketorolac, diclofenac, bromfenac), and compared these to the effects of amfenac (a metabolite of nepafenac) and to celecoxib (active principle of Celebrex). With the exception of the last compound, none of the drugs promote swelling of normal mitochondria that are well energized by succinate oxidation. However, swelling is seen when the mitochondria are under an oxidative stress due to the presence of t butylhydroperoxide. When used at 200 microM the order of potency is celecoxib > bromfenac > diclofenac > ketorolac > amfenac > nepafenac approximately equal to 0. Again with the exception of celecoxib, this swelling is not seen when mitochondria are depleted of endogenous Ca(2+) and is accelerated when exogenous Ca(2+) is provided. Sr(2+) does not substitute for exogenous Ca(2+) and prevents swelling in the presence of endogenous Ca(2+) only. The same is true for ruthenium red (inhibitor of the Ca(2+) uniporter), for cyclosporin A (inhibitor of the mitochondrial permeability transition), and for a 3.4 kDa polyethylene glycol (polymer that cancels the force which drives swelling following the permeability transition). It is concluded that several non-steroidal anti inflammatory drugs promote the mitochondrial permeability transition under conditions of oxidative stress and in a Ca(2+) dependent fashion, whereas celecoxib functions by another mechanism. Potency of those compounds that promote the transition varies widely with bromfenac being the most potent and nepafenac having almost no effect. The mitochondrial dysfunction which is caused by the transition may underlie side effects that are produced by some of these compounds. PMID- 19810215 TI - Evolution of archosaurian body plans: skeletal adaptations of an air-sac-based breathing apparatus in birds and other archosaurs. AB - Living birds represent the only extant sauropsid group in which pulmonary air sacs pneumatize the postcranial skeleton. Notable in this regard is an extraordinary degree of variability, ranging from species that are completely apneumatic to those characterized by air within the entire postcranial skeleton. Although numerous factors (e.g., body size) have been linked with "relative" pneumaticity, comparative studies examining this system remain sparse. This project sought to (1) characterize whole-body patterns of skeletal pneumaticity in distantly related neognath birds and (2) evaluate putative relationships among relative pneumaticity, body size and locomotor specializations. Pneumaticity profiles were established for 52 species representing 10 higher-level groups. Although comparisons reveal relatively conserved patterns within most lower-level clades, apparent size- and locomotor-thresholds do impart predictable deviations from the clade norm. For example, the largest flying birds (vultures, pelicans) exhibit hyperpneumaticity (i.e., pneumaticity of distal limb segments) relative to smaller members of their respective clades. In contrast, skeletal pneumaticity has been independently lost in multiple lineages of diving specialists (e.g., penguins, auks). The application of pneumaticity profiling to extinct archosaurs reveals similar trends in body size evolution, particularly when examining patterns of pneumaticity in a size-diverse assemblage of pterosaurs (flying "reptiles"). As a fundamental organizing system, skeletal pneumaticity may play a role in relaxing constraints on body size evolution by allowing volumetric increases without concomitant increases in body mass. Not only might this be critical for taxa (birds, pterosaurs) exploiting the energetically costly aerial environment, but could be beneficial for any large-bodied terrestrial vertebrates such as the dinosaurs. PMID- 19810216 TI - BSVA policy on fireworks. PMID- 19810217 TI - Variation shows community services ripe for efficiencies. PMID- 19810218 TI - PCTs gear up for pandemic. PMID- 19810219 TI - Reform: Lessons for the UK in Obama's universal plan. PMID- 19810220 TI - On auld arguments. PMID- 19810221 TI - Nobody can look the other way. PMID- 19810222 TI - Swine flu: Business as usual in a pandemic. PMID- 19810223 TI - Appropriate care: Should this patient still be in hospital? PMID- 19810224 TI - Performance: GPs in the spotlight. AB - PCTs are having to review the level of service they are getting from GP practices. The balanced scorecard tool attempts to combine quality and efficiency indicators. GPs' concerns include publication of the scorecard data without agreement by the GPs. PMID- 19810225 TI - Case study. Dirty blood. Commentary. PMID- 19810226 TI - The new normal. Avoiding financial risk and boosting transparency and governance are all emphasized in the wake of September 2008's meltdown. AB - A year after the financial sector's meltdown, hospitals are still adjusting to the economic landscape. Where once they overlooked potential balance sheet stress from debt portfolios and the riskiness of their investments, they now obsess about risk and how to minimize it. "Systems are spending a lot more time on this than they were a year ago," says Kerry Rudy, of Ziegler Capital Markets. PMID- 19810227 TI - The grumbling commences. Senate Finance bill greeted by critics from all sides. PMID- 19810228 TI - RAC anxiety. Expansion of audits in reform bill worries hospitals. PMID- 19810229 TI - First diseases, now debt. Groups blame CMS for failure of demo project. PMID- 19810230 TI - New dialysis payment plan. Providers worry about effects of bundling payments. PMID- 19810231 TI - No holding back. Report unlikely to slow demand for proton therapy. PMID- 19810232 TI - Two sides of a story. ACHE's Form 990 shows losses; group claims profit. PMID- 19810233 TI - 10 years after. On eve of To Err's anniversary, changing healthcare world becomes priority. PMID- 19810234 TI - School is never out. Learning to learn is the only way to keep up with clinical change. PMID- 19810235 TI - Overlooked ingredient. Nurses provide quality, safety expertise on boards. PMID- 19810236 TI - Crunch time. AB - After an August recess that simmered with anger toward a Democratic effort to reshape the nation's healthcare system, President Barack Obama took action to regain momentum with a speech outlining his goals for reform. Providers are anxiously watching. "We're at the point of no return. We have to do something. We're going to bankrupt our federal treasury if we don't," says William Leaver, of Iowa Health System. PMID- 19810237 TI - Adding it all up. Some caution uninsured figure could be too low. PMID- 19810238 TI - Safety first. Joint Commission's new center pinpoints leadership. PMID- 19810239 TI - Whistle-blower tactic. Lawsuits shouldn't use public sources: groups. PMID- 19810240 TI - Shifts at CHI. Belt-tightening, divestitures could be trend: expert. PMID- 19810241 TI - Universal picture presents... a counterintuitive argument for insuring all and controlling costs. PMID- 19810242 TI - Up & Comers 2009. We're in good hands. Up & Comers ready to fill gap whenever execs retire. PMID- 19810243 TI - McKinsey report--the aftermath. Politicians accused of not facing reality. PMID- 19810244 TI - London acutes could see workload fall by up to 72pc. PMID- 19810245 TI - NHS spending: McKinsey exposes hard choices to save 20bn pounds sterling. PMID- 19810247 TI - Thriving in an austere landscape. PMID- 19810246 TI - On managers fighting back. PMID- 19810248 TI - Breastfeeding: Get the genie out of baby's bottle. PMID- 19810249 TI - Cold weather planning: The flu pandemic that came in from the cold. PMID- 19810250 TI - Human resources: The rough guide to being tough. PMID- 19810251 TI - Technology: Great what you can get on eBay... PMID- 19810252 TI - Overview: The buzz starts here. PMID- 19810253 TI - The Innovation Fund: It's not just about gadgets. PMID- 19810254 TI - Main stage: Speakers' corner. PMID- 19810255 TI - Forum: Up close and personal. PMID- 19810256 TI - Showcase: Regional trailblazers. PMID- 19810257 TI - WIBGI: Want one of them. PMID- 19810258 TI - McKinsey savings report: DH is told 137,000 posts must go in next five years. PMID- 19810259 TI - Mid staffs means a new outlook for regulators. Interview by Dave West. PMID- 19810260 TI - On sacred cows. PMID- 19810262 TI - Accountability: The friendly face of scrutiny. PMID- 19810261 TI - Release the reins on your team. PMID- 19810263 TI - Swine flu: Lessons from the first wave of a pandemic. PMID- 19810264 TI - Beyond the fringe benefits. AB - NHS employers will be looking for savings on staff costs--but room to cut wages is limited. Perks such as cheap staff parking and workplace nurseries could be targeted for cuts. Trusts will be keen to avoid the debacle of 2006 when frontline staff were laid off. As NHS spending is fixed till 2011, trusts have time to prepare--and find new ways of working. PMID- 19810266 TI - PCTs must be ready to pay extra GP costs. PMID- 19810265 TI - 'Organic' reduction in PCTs under Tories. PMID- 19810267 TI - NHS IT. Control delete: a local future for IT. PMID- 19810268 TI - Getting good at governance. PMID- 19810269 TI - Diversity: Commission for all your patients. PMID- 19810270 TI - Payment by results: Get under the skin of nursing's real costs. PMID- 19810271 TI - Pride of the community. AB - District nursing is changing to reflect a modern service. The workforce is ageing and training has been slashed. Renewed commitment to the sector is necessary to deliver the vision laid out in the next stage review. PMID- 19810272 TI - Seminal papers don't age. PMID- 19810273 TI - Hidden risk of cars. PMID- 19810275 TI - Swine flu and meningitis. PMID- 19810276 TI - Abstracts of the 25th Congress of the European Committee for Treatment and Research in Multiple Sclerosis. September 9-12, 2009. Dusseldorf, Germany. PMID- 19810277 TI - Smoking and lung cancer: causality, Cornfield and an early observational meta analysis. PMID- 19810278 TI - Staying attuned to blood pressure. Blood pressure--especially that top number- should get more of our attention. PMID- 19810279 TI - Exercise to go. Away from home? You can still get a good workout, even in your hotel room. PMID- 19810280 TI - Radio wave treatment is effective against Barrett's esophagus. PMID- 19810281 TI - Pelvic organ prolapse: vaginal delivery is not the only cause. PMID- 19810282 TI - By the way, doctor. What exactly happens when I eat something cold and get an ice-cream headache? Is it harmful in any way? PMID- 19810283 TI - Transposon Tn917PF1 mutagenesis in Bacillus licheniformis. AB - The plasmid pTnPFl containing the transposon Tn917PF1 was introduced into the protoplasts of two Bacillus licheniformis strains in the presence of polyethylene glycol. Transpositions were produced at high temperature which inhibited plasmid replication and kanamycin was used for selection.Transposon Tn917PF1 was inserted randomly into the bacterial chromosome,producing different auxotrophic, prophage BLF and bacitracin-non-producing mutants. The auxotrophic mutant phenotypes were characterized by the Holliday-test and some mutations by hybridization with a transposon DNA probe. Insertions for the entire chromosome or for the prophage genophore were found at random, but preferred target sites were detected within limited regions, like the bacitracin synthetase or sulphate reductase genes. The partial physical map of the chromosomal region of bacitracin synthetase was constructed based on the hybridization patterns of insertion mutants. PMID- 19810284 TI - Comment on "An etched track detector for short-term screening measurements of radon". PMID- 19810285 TI - Comments on "Impact of tritium around EDF nuclear power plants". PMID- 19810286 TI - Comment on "Response to 'More on the risk of cancer among nuclear workers'". PMID- 19810287 TI - Comment on "Mammography-oncogenecity at low doses". PMID- 19810288 TI - Comments on "Are the risks from tritium exposures being underestimated?". PMID- 19810289 TI - Curbing teen tobacco use. PMID- 19810290 TI - Interspousal transmission of hepatitis C: concerns and facts. PMID- 19810291 TI - Interspousal transmission of hepatitis C virus. AB - OBJECTIVE: To estimate the sero-prevalence of anti-HCV among the spouses of HCV positive patients. DESIGN: A cross-sectional survey. PLACE AND DURATION OF STUDY: Shifa International Hospital, Islamabad, from January 2001 to January 2004. PATIENTS AND METHODS: Spouses of patients of hepatitis C were studied. Spouses of patients who had hepatitis C by the presence of anti-HCV and HCV RNA (PCR) were tested for anti-HCV by ELISA method. RESULTS: A total of 347 consecutive patients were included in the study. Fifty-three patients were single, 48 were widows or divorced and 19 individuals refused to be tested. Thus, 227 patients were available for anti-HCV testing. Out of these 227 spouses tested, 10 (4.4%) were found to be positive for anti-HCV. The mean age of HCV-positive spouses was 44.2 +/- 8.31 years, while that of HCV-negative spouses was 46.37 +/- 11.31 years (p = 0.55). The mean years of marriage of HCV-positive spouses was 20.1 +/- 9.79 years as compared to 21.1 +/- 9.96 years for HCV-negative spouses (p = 0.75). The male proportion of HCV-positive spouses was 60% as compared to 57.6% for HCV-negative spouses (p = 0.9). The patients and spouses had common risk factor of history of injections. CONCLUSION: A positive anti-HCV was found in 4.4% spouses and common risk factors of history of injection and transfusion were noted in the patients and spouses. PMID- 19810292 TI - Frequency of hepatorenal syndrome among cirrhotics. AB - OBJECTIVE: To determine the frequency of hepatorenal syndrome in patients with chronic liver disease and ascites. DESIGN: A cross-sectional study. PLACE AND DURATION OF STUDY: From July 2001 to March 2002 at Civil Hospital, Karachi. PATIENTS AND METHODS: The study included 240 patients who were admitted in the Medical Wards of Civil Hospital, Karachi, during the study period. The subjects were diagnosed as having chronic liver disease and all of them had ascites. All the patients with renal dysfunction were identified and were worked up for hepatorenal syndrome and other causes of renal dysfunction. RESULTS: Of two hundred and forty (240) patients enrolled in the study, 148 were male (61.7%) and 92 were female (38.3%). Seventy-six (76) patients (31.6%) were found to have renal impairment as diagnosed by serum creatinine level of 1.5 mEq/L or greater. Six (6) patients showed improvement in their serum creatinine level (below 1.5 mg/dL) after receiving 1.5 liter of normal saline infusion and diuretic withdrawal. Eleven (11) patients were diagnosed to have primary renal disease based on urine analysis, urinary protein excretion and ultrasound examination. Six (6) patients were diagnosed to have renal dysfunction secondary to analgesic nephropathy and in seventeen (17) patients renal dysfunction was secondary to spontaneous bacterial peritonitis. The remaining 36 (47.4%) patients with renal dysfunction were diagnosed as suffering from hepatorenal syndrome based on the diagnostic criteria. This represents the frequency of 15% among patients with chronic liver disease and ascites. CONCLUSION: Hepatorenal syndrome is common complication in patients with cirrhosis and ascites. It was the most common cause of renal impairment in this series followed by other causes like spontaneous bacterial peritonitis, primary renal disease, analgesic nephropathy and hypovolemia. PMID- 19810293 TI - The outcome of subtotal abdominal hysterectomy. AB - OBJECTIVE: To determine the outcome of subtotal abdominal hysterectomy in terms of intraoperative and postoperative complications in women with benign uterine conditions. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: The Jinnah Medical and Dental College and Hospital, Karachi from March, 2003 to March, 2004. PATIENTS AND METHODS: The study included 50 women, who underwent subtotal abdominal hysterectomy due to technical difficulties during surgery in benign uterine diseases. Inclusion criteria were menorrhagia and pelvic pain, dysfunctional uterine bleeding, repeated pelvic infection refractory to medical treatment, fibroid uterus and ovarian cyst with adenomyosis of uterus. Patients over 60 years of age, uterine prolapse and suspected cancer of cervix were excluded. Detailed history was taken and examination done. Routine investigations including ultrasound were carried out. Subtotal hysterectomy was performed by clamp-cut and ligate method. RESULTS: The mean age of patients undergoing hysterectomy was 44 years and parity between 4 - 10. Most common complaint was excessive menstrual loss. Fibroid was found in 10 (20%) cases, dysfuntional uterine bleeding in 15 (30%), pelvic pain and menorrhagia in 5 (10%), ovarian cyst with adenomyosis in 13 (26%) and repeated pelvic infections refractory to medical treatment in 7 (14%).There was no injury to adjacent vicera during the procedure. Early complications like temperature was noted in 5 (10%) and late complications like cyclical menstrual bleeding in 1 (2%) and vaginal discharge in 2 (4%) patients. All cases were followed-up 3 and 6 monthly and urinary, sexual and bowel functions were found to be unaffected during this period. There was no mortality associated with the procedure. The histopathology of specimen revealed fibroid followed by adenomyosis to be the commonest pathology. CONCLUSION: Subtotal abdominal hysterectomy resulted in less operative time, rapid recovery, fewer short-term complications but infrequently caused cyclical bleeding and vaginal discharge. PMID- 19810294 TI - Estrogen replacement therapy for depression in perimenopausal women. AB - OBJECTIVE: To determine the effect of a four-week course of oral estrogen therapy on depression in aging women. DESIGN: Quasi experimental. PLACE AND DURATION OF STUDY: The OPD, Bahawal Victoria Hospital, Bahawalpur, from September 2000 to January 2004. PATIENTS AND METHODS: Forty-two depressed women (aged 42-57) who were either perimenopausal (n = 20) or postmenopausal (n = 22) received open label treatment with oral estrogen tablets (1.25 mg/day) for 4 weeks. The Montgomery-Asberg Depression Rating Scale and the Beck Depression Inventory were used to assess depressive symptoms, the Greene Climacteric Scale was used to assess menopause-related symptoms, and the Clinical Global Impression(CGI) was used to assess global clinical improvement in these women at baseline and after treatment. Remission of depression was defined as a score < 10 on the Montgomery Asberg Depression Rating Scale and a score < or = 2 on the CGI at week 4. RESULTS: The women who completed the study had a median Montgomery-Asberg Depression Rating Scale score of 20 (range = 15-32) at study entry and 11.50 (range = 10-31.0) at week 4 (z = -3.43, p < 0.01). This improvement was consistent with that reported by the women themselves on the Beck Depression Inventory (rs = 0.86, n = 20, p < 0.01). The improvement measured by CGI scores was also significant (p < 0.01). Remission of depression was noted in 5 of the 40 women (after 1 year) who completed the study. Anti-depressant response was not associated with severity or subtypes of depression at study entry or with concomitant improvement in menopause-related symptoms. CONCLUSION: Perimenopausal and postmenopausal women benefit from short-term use of estrogen therapy. Antidepressant effect of estrogen therapy is independent of improvement in menopause-related symptoms. PMID- 19810295 TI - Comparison of unfractionated heparin vs low molecular weight heparin in the treatment of acute deep vein thrombosis. AB - OBJECTIVE: To compare the results of unfractionated heparin (UFH) with low molecular weight heparin in the treatment of acute deep vein thrombosis of lower limb. DESIGN: Randomized control trial. PLACE AND DURATION OF STUDY: Bolan Medical Complex Hospital, Quetta from January 2002 to July 2003. PATIENTS AND METHODS: Thirty patients who presented with acute deep vein thrombosis of lower limb, confirmed by either Doppler ultrasonography or venography, were selected for the study. Patients were divided randomly into two groups for treatment. Group 1 was started with unfractionated heparin while group 2 with low molecular weight heparin. Comparison of two treatments to determine the efficacy was done by certain criteria like pain improvement, reduction in swelling, alteration in bleeding profile, complications of therapy, recurrence, morbidity and mortality. RESULTS: The age of the patients ranged from 16-82 years. There were 15 females (50.00%) and 15 males (50.00%). Pain and swelling were present in all patients (100%), while temperature and superficial vein dilation in 43.3% and 30% respectively. The distribution of DVT in left lower limb was in 13 patients (43.33%), right lower limb involvement in 12 patients (40.00%) and both limbs involvement in 5 patients (16.67%). In group 1 improvement in pain occurred after 4th day in 13 patients, while in group 2 before 4th day in 8 patients (p-value = 0.068). Improvement in swelling was observed after 6th day in 11 patients (group 1), while before 6th day in 8 patients (group 2) (p-value = 0.171). Bleeding time was prolonged in 5 patients in group 1 and statistically found significant (p value = 0.014), while in group 2 it was normal. Thromboembolism in 3 patients and major bleeding was observed in 2 patients in group 1, while in group 2 it was normal. Recurrence was reported in 2 patients in group 1 and 1 patient in group 2 (p-value 0.0815). The hospital stay was more than 10 days in group 1 (12 patients), and less than 10 days in group 2 (13 patients). It was found statistically significant (p-value = 0.001). Three patients died in group 1, while no mortality was observed in group 2. CONCLUSION: Treatment with low molecular weight heparin (LMWH) has good patient compliance and is easy to administer. LMWH has an advantage over UFH due to its normal bleeding profile and significantly less hospital stay. PMID- 19810296 TI - Comparison of an integrated problem-based learning curriculum with the traditional discipline-based curriculum in KSA. AB - OBJECTIVE: To compare the integrated problem-based learning (PBL) curriculum of the third year medical students of KSU at the Al-Qaseem campus with the traditional discipline-based teaching curriculum of students of Riyadh campus. DESIGN: A comparative cross-sectional study. PLACE AND DURATION OF STUDY: Al Qaseem and campuses of King Saud University (KSU). A structured questionnaire was used for comparing the students of third year at both campuses of KSU. The student's learning behaviours and their liking for teaching methodologies was evaluated from March to June 2003. SUBJECTS AND METHODS: There were 138 3rd year students of Riyadh campus (59 females and 79 males) and 65, of Al-Qaseem campus (17 females and 48 males) (N = 203). Fifty-eight percent of students' at Al Qaseem campus went to the library two or more times compared to 3.4% of the Riyadh campus (P = 0.000), 18.5% of Al-Qaseem campus used Journals (P = 0.000), and 86% did self-learning (P = 0.000), which was much higher than the students' at Riyadh campus. RESULTS: Almost 75% students at Al-Qaseem campus were satisfied with their system of education compared to 20% at Riyadh campus (P = 0.000). Sixty-nine percent students at Al-Qaseem campus answered problem-solving questions correctly versus 7.25% of the students at Riyadh campus. The students of both campuses liked group teaching almost equally and thought it makes them remember better. The Al-Qaseem campus students liked PBL, clinical skills lab, tutorials, and also lectures delivered at the college. CONCLUSION: PBL improved the students' usage of learning resources, problem-solving abilities and they were satisfied with their learning and the curriculum. PMID- 19810297 TI - Brain abscess in children. AB - OBJECTIVE: To find out the clinical presentation, radiological characteristics, various underlying predisposing conditions and causative organisms of brain abscess in children in our setup. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: The Children's Hospital and the Institute of Child Health, Lahore, over two years from September 2001 to August 2003. PATIENTS AND METHODS: All children (< 16 years) presenting with brain abscess were included to study demographic, clinical and radiological features. In addition, attempts were made to find out underlying predisposing conditions and causative organisms. RESULTS: Twenty-five children with brain abscess were managed over 2 years. The mean age was 7.8 years (range 9 months to 16 years). Male to female ratio was 2.1:1. Most patients (43%) presented with 4 weeks history of illness, with mean duration of illness at presentation of 29.3 days. Main presenting complaints were fever (72%), vomiting (48%), headache (44%) and convulsions (32%). Five patients (20%) had papilledema at presentation, another 4 (16%) had paresis/paralysis and 3 (12%) had cranial nerve palsies. Majority (64%) had solitary abscess, located in parietal, temporal, frontal and occipital lobes in order of frequency. No underlying predisposing condition was identified in 8 (32%) cases; while 8 (32%) had cyanotic congenital heart disease, 5 (20%) patients had otic infection (mastoiditis), 2 (8%) were postoperative cases and one each developed brain abscess secondary to ventriculo-peritoneal (VP) shunt infection and pulmonary tuberculosis. Causative organisms were isolated in 40% cases, which included staphylococcus aureus, Staphylococcus epidermidis, Streptococcal species, Klebsiella, E. coli and Proteus. CONCLUSION: Awareness of predisposing factors, early recognition of clinical features and understanding of the prevalent microbial profile is imperative for better management of children with brain abscess. PMID- 19810298 TI - Serum zinc levels and effects of oral supplementation in pregnant women. AB - OBJECTIVE: To evaluate the effects of oral zinc supplementation on the serum zinc levels of pregnant women. DESIGN: Experimental (double blinded randomized controlled trial). PLACE AND DURATION OF STUDY: PIMS and KRL Hospital, Islamabad, and community in tehsil Kahuta from April 2003 to April 2004. PATIENTS AND METHODS: Pregnant women of 10 to 16 weeks gestation were invited to enter the study on their booking visit. A sample size of 125 in each group was calculated. After taking an informed consent, they were assigned to control or test group by simple random sampling technique. A detailed questionnaire was filled-up by trained staff and initial evaluation along with serum zinc samples was collected. The subjects were given either zinc sulphate powder, equivalent to 20 mg elemental zinc, or were given placebo treatment along with routine supplements. These patients were followed up throughout the pregnancy by health care providers and their compliance was monitored. At delivery, serum samples were again collected for zinc estimation. The data was entered on computer, cleaned and analyzed. Paired t-test was used for comparison of means. RESULTS: The data of 242 subjects was analyzed at the end of the study. The mean age of the study participants was 25.7 +/- 4.8 years (range 16 to 40). Both the groups were similar in other demographic variables as socioeconomic status, education, BMI, height and weight. One-third of the patients had serum zinc levels below 64 microg/dl at the start of the study. A 128 pairs of pre and post-serum zinc levels were analyzed in the two groups (64 pairs in each group) to compare the means. The zinc supplemented women showed a mean increase of 14.7 microg/dl (95% CI 5-23) (P = 0.002). On the other hand the non-supplemented group showed an actual decrease in the serum zinc level which, however, did not reach statistical significance (P = 0.47). CONCLUSION: Oral zinc supplementation of pregnant women with 20 mg elemental zinc was effective in raising the serum levels of zinc. It is suggested that supplementation trials with larger dose of zinc should be carried out. PMID- 19810299 TI - Outcome of mechanical mode of induction in failed primary labor induction. AB - OBJECTIVE: To determine the role of an alternate (mechanical) inducing agent in failed labor induction. DESIGN: Quasi-experimental study. PLACE AND DURATION OF STUDY: Labor Ward at Mother and Child Health Center (MCH), Pakistan Institute of Medical Sciences (PIMS) from May 1999 to July 2003. PATIENTS AND METHODS: Women with singleton term gestation and normal live fetus undergoing indicated induction of labor were included if Bishop score was < or = 5 after 12-18 hours of primary induction with PGE2 vaginal tablets. They were then induced with mechanical method i.e. intracervical foley catheter, if fetal status was reassuring and no evidence of labor onset. The main outcome measure was mode of delivery. Secondary outcome measures were induction labor interval and induction delivery interval after second mode of induction and neonatal Apgar score. RESULTS: Thirty-six women received a second mode of induction for failed induction. Four patients were excluded. Of 32 eligible women, 24 (75%) delivered vaginally. Eight patients (25%) had emergency caesarean section. Mean induction labor interval after second mode of induction was 6.8 hours and mean induction delivery interval was 12.39 hours. Mean Apgar score at 1 minute and 5 minutes was similar in both vaginal deliveries and caesarean sections. CONCLUSION: When an alternate mechanical method was used, 75% of women labeled as "failed induction" delivered vaginally. PMID- 19810300 TI - Comparison of condylar positions in centric relation and centric occlusion in pre treatment malocclusion cases. AB - OBJECTIVE: To compare the centric relation-centric occlusion discrepancy in Angle's class II and class III with Angle's class I malocclusion cases in sagittal (antero-posterior) plane as determined by the difference in the overjet. DESIGN: Comparative; cross-sectional study. PLACE AND DURATION OF STUDY: Orthodontic Outpatient Department, Karachi Medical and Dental College, from June 2003-August 2004. PATIENTS AND METHODS: Patients were divided into 3 groups according to the Angle's classification. Leaf gauges were employed for muscle deprogramming. Bimanual manipulation technique was used to guide the mandible in to centric relation. Softened red modeling wax was used to obtain the centric bite registration. The study casts were trimmed in centric relation using bite registration. Overjet was recorded in centric relation and centric occlusion by dial caliper on casts. RESULTS: The overjet values for centric relation-centric occlusion discrepancy were highly significant for Angle's class II (p-value 0.002) and Angle's class III (p-value 0.005) when compared with Angle's class I malocclusion. CONCLUSION: Centric relation-centric occlusion discrepancy can occur in any dental class regardless of age and gender. However, especial emphasis should be given to Angle's class II and III (with functional shift) cases when evaluating any orthodontic patient. PMID- 19810301 TI - Eclampsia--management and outcome with magnesium sulphate as the anticonvulsant. AB - OBJECTIVE: To study the maternal outcome in terms of morbidity and mortality with the use of magnesium sulphate in eclampsia for seizure control and prophylaxis. DESIGN: A quasi experimental study. PLACE AND DURATION OF STUDY: This study was conducted in Obstetrics and Gynaecology Unit of Hayatabad Medical Complex, Peshawar, from 1st September 2002 to 31st December 2003. PATIENTS AND METHODS: All the patients admitted with eclampsia during this period were included in the study. A detailed history and clinical examination were recorded. Patients were monitored for number of fits, rise in blood pressure (BP), dose of magnesium sulphate administered and its side effects, development of complications of eclampsia, duration of labour, mode of delivery and use of other medications as well as maternal and perinatal outcome. RESULTS: During this period, 52 patients were admitted with eclampsia. Majority were unbooked. Primigravida were 31 (59.6%). Common age group was between 21 and 30 years having 34 (65.4%) patients while 16 (30.7%) patients were 20 years or less than 20 years of age. Thirty three (63.4%) patients had antenatal fits, 9 (17.3%) intrapartum fits and 11 (19.2%) had postpartum eclampsia. Twenty-six patients had term pregnancy, 22 patients with 28-36 weeks gestation while 4 patients developing eclampsia before 28 weeks gestation. Eighteen patients had spontaneous vaginal delivery; labour was induced in 21 patients while 6 patients underwent cesarean section. Maternal complications included pulmonary oedema, tongue bites, HELLP syndrome, placental abruption and coagulopathy. Side effects of magnesium sulphate were low. There were 4 (7.6%) maternal deaths due to eclampsia. Perinatal outcome was good with 33 (63.4%) live births, 3 IUDs and 10 still births, mostly due to prematurity. CONCLUSION: Magnesium sulphate was found to be an effective anti-convulsant both for control and prophylaxis of seizures in eclampsia and improved maternal and fetal outcome. PMID- 19810302 TI - Management of idiopathic rectal prolapse in children. AB - OBJECTIVE: To determine the percentage of patients of idiopathic rectal prolapse improving on spontaneously over the period of observation (phase I), and to determine the outcome of patients with rectal prolapse who received injection sclerotherapy (phase II). DESIGN: Quasi-experimental study. PLACE AND DURATION OF STUDY: National Institute of Child Health, Karachi from April 2001 to March 2002. PATIENTS AND METHODS: The study was conducted in two phases. In phase I of the study, newly diagnosed patients of idiopathic rectal prolapse were followed without any treatment, till the spontaneous resolution of rectal prolapse. The time period at which 50% patients improved clinically was called 'time for spontaneous resolution 50%, (TSR 50%). In phase II, injection sclerotherapy (IST) was given to those patients whose prolapse was of more than three months duration. This was a separate cohort of patients. RESULTS: One hundred patients were inducted in the study. They all had idiopathic rectal prolapse and their ages ranged from 6 months to 12 years with mean age of 5.30 +/- 2.30 years. In phase I, out of a total 50 patients, 40 could be followed with non-interventional strategy. In more than 50% of patients, the prolapse disappeared within 3 months. This was called TSR 50%. In phase II study, out of 50 patients who received IST, 29 improved within 2 weeks of single injection while 12 more improved with second injection within two months. Overall rate of resolution of prolapse at two months (41/50) was highly significant in comparison with proportion of improvement in phase I patients with p-value of 0.001. Four patients received third injection. At the end of three months prolapse disappeared in all patients of this phase (p value < 0.0001). No complication related to injection occurred. CONCLUSION: Both non-operative and injection sclerotherapy are effective in managing idiopathic rectal prolapse in paediatric population, but in terms of early recovery injection sclerotherapy is recommended as it is associated with less morbidity and is cost-effective. PMID- 19810303 TI - Management of postburn digital flexion contractures. AB - OBJECTIVE: To determine the outcome of plantar/palmer skin graft for the management of postburn digital flexion contracture in terms of functional and cosmetic improvements. DESIGN: Interventional quasi-experimental study. PLACE AND DURATION OF STUDY: Department of Plastic and Reconstructive Surgery, PGMI, Hayatabad Medical Complex, Peshawar. The duration of the study was 1-1/2 years (from January 2001 to June 2002). PATIENTS AND METHODS: Fifty patients with postburn digital flexion contracture were recruited in the study. Patients with posttraumatic and postinfective contractures were excluded from the study population. The contracture was released and the deficit produced was replaced by plantar/palmer split skin graft. Active range of motion (ROM) was measured before and after surgery. Postoperative follow-up was done for one year. The outcome measurements were color and texture match of graft with the adjacent palmer skin, sensibility, recurrence rate of contracture and donor site morbidity. The data collected was labeled with the help of statistical package for social sciences (SPSS) and descriptive statistics were calculated accordingly. RESULTS: The graft take was complete (100%) in 48 patients, 50% in one patient (2%) and nil in one (2%) patient. Recurrence of contracture occurred in 5 patients (10%). Pre operative median active range of motion (ROM) at proximal interphalangeal (PIP) joint was 60 degree to 90 degree (extension/flexion). Postoperatively median active range of motion (ROM) at PIP joint was 0 degree to 90 degree at one year follow-up examination. The healed graft showed good colour and texture match with the surrounding palmer skin. The sensory function return was satisfactory and there was no significant donor site morbidity. CONCLUSION: The excellent colour, texture match and the functional advantages in terms of active range of motion and sensibility offered by the split thickness palmer/plantar skin grafts exceeded the expected outcome of conventional techniques (thigh, groin and buttock skin grafts). PMID- 19810304 TI - An analysis of force between attractive magnets used in orthodontics. AB - OBJECTIVE: To compare the results of hard and soft attractive magnet combinations to the gold standard Neodymium Iron Boron Magnets used for extrusion of impacted maxillary canines. DESIGN: Experimental and comparative study. PLACE AND DURATION OF STUDY: Eastman Dental Institute and Hospital, London, UK in collaboration with Department of Physics and Astronomy University College of London (UCL) during 1996-1998. MATERIAL AND METHODS: In vitro experiments with combinations of hard and soft magnets of different morphologies. Load Cell Transducer was used to record the force levels in grams with magnets placed in different vertical, horizontal and angular offsets. RESULTS: Three-dimensional mesh plots were created to visualize force level decay with increasing distance. The results were compared with previous study on hard magnet combinations only and observed for "edge effect". The present study force levels were very low albeit the edge effect was negligible. CONCLUSION: Although the edge effect phenomenon present with powerful hard magnets was minimal or nearly absent with the hard and soft magnet combination--the overall force levels were too low to be of any use in clinical scenario for extrusion of deep impacted canines. PMID- 19810305 TI - Extracorporeal shock wave lithotripsy. AB - OBJECTIVE: To determine the outcome of patients undergoing extracorporeal shock wave lithotripsy (ESWL) for treatment of upper urinary tract calculi (renal and ureteric), and to note role of double-J (DJ) stents in these patients. DESIGN: A cross-sectional analytical study. PLACE AND DURATION OF STUDY: Department of Urology, Rawalpindi General Hospital, Rawalpindi, from February 1999 to July 2001. PATIENTS AND METHODS: Record of patients who underwent ESWL for renal and ureteric stones was retrieved and analyzed using statistical program, SPSS version-10 and Epi-Info 2000. In some patients pre-ESWL DJ stents were placed because of various reasons like solitary kidney, large stone volume etc. Patients were divided in two groups, Group I, in whom DJ stents were not placed, and Group II, in whom DJ stents were placed. ESWL was performed in each subject in standard way employing piezoelectric lithotripter E.D.A.P. LT 02X. Patients were evaluated for stone clearance fortnightly with X-ray or ultrasound. RESULTS: Four hundred and thirty-two patients, 68.8% male and 31.2% female, underwent ESWL. Mean age of patients was 37.7 +/- 13.1 years. Majority of patients (78.47%, n = 339) had renal, while rest had ureteric stones. Group I and II included 408 (94.4%) and 24 (5.6%) patients respectively. Renal stones were present in 78% (n=318) of Group I and 87.5% [n = 21] of Group II patients. Mean size of stones in Group I and II patients was 10.91 +/- 4.6, and 10.4 +/- 4.7mm. Stone clearance was 96.3% and 100% in Group I and Group II patients respectively. Significantly more ESWL sessions were required for stone clearance in Group II (p-value 0.03); in addition Group II patients had significantly more complications (p-value 0.01). CONCLUSION: ESWL is an effective procedure. Pre-ESWL stenting is associated with increased numbers of ESWL sessions and more complications. PMID- 19810306 TI - Greek technique for type 4a coronary artery bifurcation lesions. AB - In routine coronary angiography, bifurcation lesion is not uncommon. Current practice of dealing with type 4a coronary bifurcation lesions (lesions of main branch without significant lesions of the side branch) may lead to true bifurcation lesions after stenting due to axial plaque redistribution. This series describes an experience with Greek technique for treatment of type 4a bifurcation lesions in 18 patients for primary stenting of main vessel with simultaneous kissing balloon of side branch in an effort to avoid snow plough effect. PMID- 19810307 TI - Surgical management and outcome in achalasia cardia. AB - The objective of the evidence based clinical report was to assess the outcome of exramucosal modified Heller's cardiomyotomy in terms of effective palliation of dysphagia with minimum complications. Ten patients were included after exclusion of four patients due to poor follow-up. Due to the non-availability of manometry, barium swallow and endoscopy had to be relied upon for the definitive diagnosis. All patients underwent transabdominal modified Heller's myotomy coupled with anterior partial fundoplication (Dor Patch). Median age of the patients treated was 42.5 years. Seven patients had grade III dysphagia, 2 had grade IV and one patient, an 8 years old child was having grade II dysphagia. In one patient, (10%), mucosa was perforated iatrogenically followed by wound infection. Another patient developed acute coronary syndrome on first postoperative day. In the rest of patients recovery was uneventful. Mean hospital stay was 7.1 days. Immediate relief of dysphagia was noted in all the patients. Mean follow-up was 7.2 months. There was significant improvement in dysphagia following surgery. Outcome was assessed devising a scoring system. Excellent outcome was observed in 80% of cases, good in 10% and fair in 10% of cases. PMID- 19810308 TI - Continuity of patient care among family practice patients at a teaching hospital. PMID- 19810309 TI - Late in-stent restenosis in a patient with drug-eluting stent. AB - Drug coated stents have reduced the incidence of in-stent restenosis with coronary intervention. Whether this effect is long lasting or just delaying the process is not clear. We report here a case of late in-stent restenosis with Rapamycin drug eluting stent. PMID- 19810310 TI - Pseudoxanthoma elasticum. AB - Pseudoxanthoma elasticum is an inherited disorder of connective tissue that is associated with numerous systemic manifestations including various cardiovascular abnormalities. We present a case of a 35 years old lady who presented with abnormal wrinkling, pigmentation and laxity of skin. She also had hypertension, peripheral vascular disease, angioid streaks and mitral valve prolapse. She was thoroughly investigated and managed accordingly. She was also educated to minimize the potential risks involved and advised regular follow-up. PMID- 19810311 TI - Male pseudohermaphroditism. AB - An unusual case of a 40-year-old infertile male is presented. On examination he had cryptorchidism with other normal secondary sexual characters. Ultrasonography and computerized tomography raised the suspicion of tumour in the right abdominal testis. On exploration, a uterus with cervix, fallopian tubes and abdominal testes were detected. This was confirmed histopathologically. Chromosomal analysis revealed 46XY. PMID- 19810312 TI - Leiomyosarcoma of femoral vein in an adult. AB - A middle aged female presented with a swelling in the groin on the left side. On examination, it was an oval mildly tender firm mass, adherent to underlying structures and extending into the pelvis with pulsatility and bruit. It was diagnosed as leiomyosarcoma of femoral vein on fine needle aspiration cytology and selective left common iliac and common femoral angiography of the lesion. En bloc wide excision of the mass including the femoral vessels was carried out with saphenous vein graft interposition for restoration of vascular continuity for both vessels. Femoral nerve was sacrificed for tissue clearance. The defect was large and led to permanent limp and walk with a support. The lady was followed up for two years. She was found to have no local recurrence or detectable metastases. PMID- 19810313 TI - Perimalleolar ulcers in hydroxyurea treated patients with concomitant chronic venous disease: diagnostic pitfalls. AB - We report three male patients, with duplex confirmed chronic venous disease, who were on treatment with hydroxyurea for chronic myelogenous leukaemia (CML) and polycythemia vera (PV), referred to us for the management of non-healing perimalleolar ulcers of varying durations. Treatment of the ulcers, based on standard venous ulcer treatment protocols, proved futile, and one patient actually underwent surgery for varicose veins. The ulcers healed only after the discontinuation of hydroxyurea. In another patient, the ulcer continued to progress along with emergence of ankle oedema, venous eczema and lipodermatosclerosis, even after discontinuation of hydroxyurea. The short duration of treatment with the drug before the onset of ulcer and characteristic skin changes of chronic venous disease (CVD), in this patient, made us reconsider our diagnosis and this patient ultimately benefited from varicose veins surgery. Based on our experience with these cases, we have made an attempt to differentiate between the chronic venous ulcer and the drug-induced ulcer on clinical grounds. PMID- 19810314 TI - Concurrent fibroadenoma of breast and fibromatosis of vulva and thigh. AB - Fibromatosis is a rare locally aggressive fibroproliferative neoplasm. Vulvar fibromatosis is extremely rare tumor. A case of a young woman is reported who presented with large vulvar, thigh and breast masses and was surgically treated. The pathological examination showed fibromatosis of vulvar and thigh masses and fibroadenoma of breast lump. PMID- 19810315 TI - An adult with Morquio syndrome. AB - We report a case of a 25-year-old male who came to endocrine clinic to get medical certificate so as to claim as the shortest man of the world. He was evaluated for short stature and eventually diagnosed with type-IV mucopolysaccharidosis (MPS) or Morquio syndrome. PMID- 19810316 TI - Low dose aspirin depresses human platelet aggregation triggered by activating agents whose mechanisms involve arachidonic acid. PMID- 19810317 TI - Direct observation of size dependent activation of NO on gold clusters. AB - Vibrational spectra of NO ligands adsorbed on cationic gold clusters are obtained using IR multiphoton dissociation spectroscopy. We observe a strong oscillation of the NO stretching frequency with increasing cluster size that reveals a clear odd-even dependence in the strength of the NO bond. For the corresponding CO complexes, no oscillation is found. This behaviour can be understood on the basis of the character and occupation of the frontier orbitals of the complexes. PMID- 19810318 TI - Parallel tempering: theory, applications, and new perspectives. AB - We review the history of the parallel tempering simulation method. From its origins in data analysis, the parallel tempering method has become a standard workhorse of physicochemical simulations. We discuss the theory behind the method and its various generalizations. We mention a selected set of the many applications that have become possible with the introduction of parallel tempering, and we suggest several promising avenues for future research. PMID- 19810319 TI - A short-range gradient-corrected density functional in long-range coupled-cluster calculations for rare gas dimers. AB - A previously proposed scheme for coupling short-range (sr) density functionals with wavefunction-based long-range (lr) ab initio methods has been extended by (a) developing a new gradient-corrected sr functional of the Perdew-Burke Ernzerhof (PBE) type and (b) introducing coupled-cluster (CC) approaches (CC with single and double excitations (CCSD), and with additional perturbative triples (CCSD(T))) at the ab initio side. The results show that mixing-in of lr-ab initio correlation helps to remove deficiencies of currently used density functionals for the treatment of van-der-Waals interactions. Compared to full ab initio calculations, the basis set dependence is weaker so that the accuracy of the mixed results surpasses that of the ab initio ones for basis sets of triple-zeta quality. PMID- 19810320 TI - Ab initio calculation of electronic absorption spectra and ionization potentials of C3H3 radicals. AB - C3H3 isomers play a major role in combustion chemistry and are of fundamental interest in understanding photodissociation processes of hydrocarbons. Nevertheless, their electronic and ionization spectra are largely unknown. This study presents the first extended investigation of the electronically excited states and ionization potentials of the four most stable isomers, namely propargyl, 1-propynyl, cycloprop-2-enyl, and cycloprop-1-enyl. Electronic spectra are studied by ab initio calculations of the vertical excitation energies at the multi-reference configuration interaction (MRCI) level of theory. Vertical and adiabatic ionization energies are determined by coupled-cluster (RCCSD(T)) and multi-reference perturbation theory (CASPT2). The obtained results are suitable to aid the experimental detection of the different C3H3 isomers. Most of all, the calculations show that the assignment of an experimentally observed absorption at 242 nm to the propargyl radical was erroneous. Therefore, some recent experiments, based on this assignment, also need to be re-interpreted because this absorption is due to a yet unknown species. PMID- 19810321 TI - Ab initio study of temporary anions of benzene and fluorobenzenes using the multipartitioning many-body perturbation theory. AB - We present an ab initio study of the lowest states of five temporary anions: C6H6(-), C6H5F(-), 1,4-C6H4F2(-), 1,2,3-C6H3F3(-), and 1,3,5-C6H3F3(-). Vertical positions and widths of anionic resonances have been calculated within the stabilization graph approach using the multipartitioning form of the many-body perturbation theory for state-selective effective Hamiltonians restricted to second order (MPPT-R). Good agreement with experimentally derived estimates justifies application of the MPPT-R method for theoretical investigation of haloaromatic temporary anion radicals. PMID- 19810322 TI - Multireference calculations of the fluorescence, phosphorescence and photodissociation of p-chlorotoluene. AB - Equilibrium geometries and vibrational frequencies of the ground and some excited states of p-chlorotoluene were calculated by the complete active space self consistent field (CASSCF) method. Multi-reference CASSCF second order perturbation theory (MSCASPT2) calculations were performed on the vertical excitation energies of six singlet and triplet excited states. The potential energy curves along the Cl-C6H4CH3 bond distance of a number of low-lying singlet and triplet excited states were calculated by the CASPT2 method based on CASSCF partially optimized geometries. The fluorescence and one component of the dual phosphorescence observed experimentally were clearly explained by the CASPT2 calculated transition energies. According to those CASPT2 potential energy curves, the photodissociation of p-chlorotoluene at 266 nm was attributed to the predissociation of the first triplet excited state after its intersystem crossing with the first singlet excited state. The internal rotation and substitution effects of methyl on the photodissociation were discussed in detail. PMID- 19810323 TI - Bending effect on the stabilities of quadruple hydrogen bonding systems: theoretical study of a series of self-constituted quadruple hydrogen bonded complexes (C9H9N5O2)2. AB - A series of self-constituted quadruple hydrogen bonded (QHB) complexes (C9H9N5O2)2 has been designed and studied systematically using density functional theory (B3LYP/6-31G**) and the Morokuma energy decompose method (HF/6-31G**). Despite very similar structures of these systems, the interaction energies fluctuate significantly from 22.33 to 88.30 kcal mol(-1). To explain this somewhat unexpected observation, several doubly hydrogen bonded (DHB) systems were designed and a "bending effect" hypothesis was presented. According to the hypothesis, the spatial arrangement of hydrogen bonds is less important than their intensity arrangement. PMID- 19810324 TI - Theoretical maximal storage of hydrogen in zeolitic frameworks. AB - Physisorption and encapsulation of molecular hydrogen in tailored microporous materials are two of the options for hydrogen storage. Among these materials, zeolites have been widely investigated. In these materials, the attained storage capacities vary widely with structure and composition, leading to the expectation that materials with improved binding sites, together with lighter frameworks, may represent efficient storage materials. In this work, we address the problem of the determination of the maximum amount of molecular hydrogen which could, in principle, be stored in a given zeolitic framework, as limited by the size, structure and flexibility of its pore system. To this end, the progressive filling with H2 of 12 purely siliceous models of common zeolite frameworks has been simulated by means of classical molecular mechanics. By monitoring the variation of cell parameters upon progressive filling of the pores, conclusions are drawn regarding the maximum storage capacity of each framework and, more generally, on framework flexibility. The flexible non-pentasils RHO, FAU, KFI, LTA and CHA display the highest maximal capacities, ranging between 2.86-2.65 mass%, well below the targets set for automotive applications but still in an interesting range. The predicted maximal storage capacities correlate well with experimental results obtained at low temperature. The technique is easily extendable to any other microporous structure, and it can provide a method for the screening of hypothetical new materials for hydrogen storage applications. PMID- 19810325 TI - Missing levels lead to additional lines: the influence of nuclear spin statistics on femtosecond degenerate four wave mixing spectroscopy of polyatomic systems. AB - We present the results of comprehensive experimental and theoretical studies of the effect of nuclear spin statistics on the structure of rotational recurrences in femtosecond degenerate four-wave mixing (fs DFWM) spectra of asymmetric top molecules. New recurrences of J-type with a period of 1/[4(B + C)] are identified in fs DFWM spectra of near-prolate tops SO2 [H. M. Frey, P. Beaud, T. Gerber, B. Mischler, P. P. Radi and A. P. Tzannis, J. Raman Spectrosc., 2000, 31, 71] and NO2 [I. Pastirk, M. Comstock and M. Dantus, Chem. Phys. Lett., 2001, 349, 71]. Our measurements of the asymmetric oblate-like top CH3NO2 revealed new recurrences of A-type with a period of 1/(8A), A, B and C being the rotational constants.The incorporation of nuclear spin statistics into the fs DFWM simulation-fitting code allowed us to successfully reproduce all these new features and the subsequent theoretical analyses uncovered mechanisms of their appearance. The nuclear spin statistics is shown to have a profound effect on fs DFWM spectra of asymmetric top species, notably with several zero spin nuclei. PMID- 19810326 TI - Ultrafast dynamics of excess electrons in molten salts: part II. Femtosecond investigations of Na-NaBr and Na-NaI melts. AB - Ultrafast dynamics of excess electrons in Na-NaBr and Na-NaI molten solutions at elevated temperatures (T = 953-1128 K) were investigated over an extended wavelength range. Modelling the time profiles resulted in two time constants tau 1 = (200 +/- 40) fs and tau 2 = (2.8 +/- 0.4) ps for both systems at 1073 K. All transients can be understood in terms of dynamical equilibria between polaron and Drude-type electrons as well as polaron and Drude-type electron forming bipolarons. In agreement with our earlier results for K-KCl melts the fast component is assigned to the relaxation of Drude-type electrons into polarons while the longer component, tau 2, represents the time during which Drude-type electrons recombine with polarons leading to bipolarons. In addition, the temperature dependence was studied in Na-Nal: Decreasing the temperature to 953 K resulted in an increase of the time constants to tau 1 = (360 +/- 50) fs and tau 2 = (4.3 +/- 0.7) ps, respectively. At temperatures, where the ionic diffusion in Na-NaI melts becomes comparable to Na-NaBr melts, the time constants for the relaxation processes also coincide. The temperature-dependent investigations resulted in an Arrhenius activation energy of (25 +/- 5) kJ mol(-1) for Na-NaI melts in good agreement with literature data. PMID- 19810327 TI - Uptake of Fe, Na and K atoms on low-temperature ice: implications for metal atom scavenging in the vicinity of polar mesospheric clouds. AB - Ice clouds form in the mesosphere between 80 and 90 km, at high latitudes during summer when the temperature falls below 150 K. There is strong evidence that the water-ice particles in these clouds scavenge metal atoms that are produced in the mesosphere by meteoric ablation. In the present study the uptake of Fe, Na and K on an ice film was studied in a fast flow tube over a temperature range of 80-150 K, covering the temperatures over which ice clouds form in the upper mesosphere. The uptake was found to be highly efficient and mostly in the diffusion-limited regime, requiring accurate measurements of the diffusion coefficients of the metal atoms in He: DFeHe = 366 (+/- 17) (T/296 K)(1.85 +/- 0.07), DNaHe 286 (+/- 13) (T/296 K)(1.68 +/- 0.04) and DKHe = 247 (+/- 15) (T/296 K)(1.69 +/- 0.07) Torr cm2 s(-1). Measured values of the diffusion coefficients in N2 are 112 (+/- 4), 125 (+/- 4) and 88 (+/- 4) Torr cm2 s(-1) at 293 K for Fe, Na and K, respectively. The uptake of Na and K was observed to be extremely efficient from 80-150 K, with lower limits of gamma Na > 0.09 and gamma K > 0.05, although it is likely that gamma is much closer to unity. The uptake of Fe on cubic ice is close to unity efficiency above 135 K, but gamma Fe decreases to only 3 x 10(-3) at 80 K. Uptake of Fe on amorphous ice films is much more efficient than on cubic ice films below 130 K. These results are interpreted using quantum calculations of the metal atoms adsorbed onto a 12-H2O model ice surface. Finally, it is shown that the uptake of Fe, Na and K on low-temperature ice is sufficiently fast to explain the substantial depletions in the mesospheric metal layers that are observed in the presence of mesospheric ice clouds. PMID- 19810328 TI - Mechanistic and kinetic study of the O + CH3OCH2 reaction and the unimolecular decomposition of CH3OCH2O. AB - Potential energy surface for the O + CH3OCH2 reaction is calculated using the coupled cluster theory with single, double, and non-iterative triple substitutions [CCSD(T)] with a complete basis set extrapolation. It is revealed that the reaction of O with CH3OCH2 proceeds dominantly via an addition/elimination mechanism. Other minor mechanisms include direct hydrogen abstraction, which may play a significant role at high temperatures, and a high barrier S(N)2 displacement. The initial adduct is the CH3OCH2O radical, which has many product channels via decomposition and isomerization. It is confirmed that beta-CH bond cleavage is the dominant product channel and all the remaining processes are of marginal significance. The rate coefficients for the barrierless association of O with CH3OCH2 are calculated to be in the range of (2.23-0.86) x 10(-10) cm3 molecule(-1) s(-1) with an apparently negative temperature dependence from 200-2000 K. For the H-atom production channel, the tunneling effect plays an important role and the barrier height is estimated to be 14.2 kcal mol(-1). PMID- 19810329 TI - Identification of possible Lewis acid sites on the beta-AlF3(100) surface: an ab initio total energy study. AB - Strong Lewis acid catalysts are widely used in a variety of industrial processes including Cl/F exchange reactions. Aluminium fluorides (AlF3) have great potential for use in such reactions. Despite the importance of the surface in the catalytic process little is known about the detailed atomic scale structure of AlF3 surfaces. In the current study we employ state of the art total energy calculations based on hybrid-exchange density functional theory to predict the composition and structure of the (100) surface of beta-AlF3 for the first time. We examine six possible terminations of the beta-AlF3 (100) surface and demonstrate that there are two relatively low energy terminations that result in the formation of under co-ordinated Al3+ ions at the surface. Such under co ordinated ions are expected to be strong Lewis acid sites. This is the first ab initio determination of the atomic scale structure of such sites on the surface of beta-AlF3. PMID- 19810330 TI - Adsorption of triblock copolymers and their homopolymers at laponite clay/solution interface. Role played by the copolymer nature. AB - The adsorption thermodynamics of copolymers, based on ethylene oxide (EO) and propylene oxide (PO) units, at the laponite (RD) clay/liquid interface was determined at 298 K. The copolymer nature was tuned at molecular level by changing the hydrophilicity, the architecture and the molecular weight (Mw) keeping constant the EO/PO ratio. Polyethylene (PEGs) and polypropylene (PPGs) glycols with varying Mw and their mixture were also investigated to discriminate the role of the EO and the PO segments in the adsorption process. Enthalpies of transfer of RD, at fixed concentration, from water to the aqueous macromolecule solutions as functions of the macromolecule molality were determined. They were treated quantitatively by means of a model based on two equilibria: (1) one-to one binding between the macromolecule and the site on the solid and (2) two-to one binding following which one macromolecule interacts with another one adsorbed onto the solid. The good agreement between the equilibrium constants obtained from calorimetry and those determined from kinetic experiments confirmed the reliability of the experimental and theoretical approaches. Almost all of the systems investigated are highlighted by the one-to-one binding; the L35 and 10R5 systems present both equilibria. The insights provided by the thermodynamics of adsorption of their homopolymers onto RD were fruitful in obtaining detailed information on the nature of the forces involved between RD and the copolymers. The data obtained in the present work clearly evidenced that for comparable polymer Mw, PPG is more suitable in building up a steric barrier around the RD particles and, indeed, exhibits several advantages and no drawbacks. Moreover, the parent copolymers may properly functionalize the RD surface by exploiting both their high affinity to the solid surface and the ability to self-assemble onto it as L35 and 10R5 clearly showed. PMID- 19810331 TI - Binding of a chiral drug to a protein: an investigation of the 2-(3 benzoylphenyl)propionic acid/bovine serum albumin system by circular dichroism and fluorescence. AB - A combined approach using global analysis of circular dichroism multiwavelength data and time resolved fluorescence was applied to investigate the interaction of R-(-)- and S-(+)-ketoprofen with bovine serum albumin in buffer solution at neutral pH. A characterization of the most stable drug : protein adducts of 1 : 1 and 2 : 1 stoichiometry, as individual chemical species, was obtained. The stability constants and the absolute circular dichroism spectra of the diastereomeric complexes were determined. The spectra of the 1 : 1 conjugates are opposite in sign, those of the 2 : 1 complexes are both negative, but different in shape from each other (peaks at 358 and 342 nm for S-(+)- and R-(-) ketoprofen, respectively). A tryptophan residue was shown to be involved in the binding of the drug, in the primary site for the R-(-) and in the secondary site for the S-(+) enantiomer, thereby showing that chiral recognition by the protein causes the site of highest affinity being not the same for both optical antipodes. PMID- 19810332 TI - Energy dissipation in slipping biological pumps. AB - We describe active transport in slipping biological pumps, using mesoscopic nonequilibrium thermodynamics. The pump operation is characterised by its stochastic nature and energy dissipation. We show how heating as well as cooling effects can be associated with pump operation. We use as an example the well studied active transport of Ca2+ across a biological membrane by means of its ATPase, and use published data to find values for the transport coefficients of the pump under various conditions. Most of the transport coefficients of the pump, including those that relate ATP hydrolysis or synthesis to thermal effects, are estimated. This can give a quantitative description of thermogenesis. We show by calculation that all of these coupling coefficients are significant. PMID- 19810333 TI - Reverse hydrogen spillover in supported subnanosize clusters of the metals of groups 8 to 11. A computational model study. AB - In a recent computational study [G. N. Vayssilov, B. C. Gates and N. Rosch, Angew. Chem., Int. Ed. Eng., 2003, 42, 1391], we found zeolite-supported Rh6 clusters, interacting with hydroxyl groups of the support, to undergo partial oxidation due to reverse spillover of hydrogen onto the metal cluster. Now, we have extended this model study to transition metal clusters M6 of the platinum and gold groups. According to the model calculations, reverse spillover of hydrogen onto the zeolite-supported metal clusters is energetically favored for all 12 metals. For most metals, the clusters M6 exhibit a compact form in either of the two states--bare supported and with hydrogen impurities. However, for Cu and Ag, the structures of the clusters with H impurities were determined to be more open, whereas Au6 exhibited an almost planar structure in either state. The estimated energy for reverse hydrogen spillover is lowest for the clusters Au6 and Ag6, 18 and 52 kJ mol(-1) per transferred hydrogen, and highest for the clusters Ir6 and Os6, 229 and 247 kJ mol(-1), respectively. Because of these model results, one would expect small metal clusters, supported on OH covered surfaces, likely to be oxidized and partially covered by hydrogen, substantially affecting the electron distribution and the chemical reactivity of the clusters. To assist in the experimental discrimination of hydrogen impurities of adsorbed metal clusters, we propose two criteria: metal core levels are predicted to be stabilized in the case of reverse hydrogen spillover and the number of metal oxygen contacts is calculated to be twice as large in clusters with hydrogen impurities. PMID- 19810334 TI - [It is time for the National Registry of Acute Coronary Syndrome]. PMID- 19810335 TI - [What should we know analyzing outcomes of registry of acute coronary events?]. PMID- 19810336 TI - [Project, logistics and methodology of the National Registry of Acute Coronary Syndrome(PL-ACS)]. PMID- 19810337 TI - [Epidemiology, treatment and prognosis of acute coronary syndrome in Silesia. Outcomes of pilot project of the National Registry of Acute Coronary Syndrome PL ACS]. PMID- 19810338 TI - [Acute coronary syndrome in Opole region. Data from the National Registry of Acute Coronary Syndrome PL-ACS]. PMID- 19810339 TI - [Treatment results of patients with acute coronary syndrome. Data form the National Registry of Acute Coronary Syndrome PL-ACS]. PMID- 19810340 TI - [Myocardial infarction with non-ST-segment elevation. Data from the the National Registry of Acute Coronary Syndrome PL-ACS in Silesia]. PMID- 19810341 TI - [Treatment results of myocardial infarction with ST-segment elevation. Data from the the National Registry of Acute Coronary Syndrome PL-ACS in Silesia]. PMID- 19810342 TI - [Treatment results of myocardial infarction with ST-segment elevation complicated by cardiogenic shock. Data from the the National Registry of Acute Coronary Syndrome PL-ACS in Silesia]. PMID- 19810343 TI - [Surgical revascularization in acute coronary syndrome. Data from the the National Registry of Acute Coronary Syndrome PL-ACS in Silesia]. PMID- 19810344 TI - [Female-male. Can they be treated the the same way? Primary analysis of data from the National Registry of Acute Coronary Syndrome PL-ACS]. PMID- 19810345 TI - Field note: hydraulic containment of a BTEX plume using poplar trees. AB - In 1999, 275 poplar trees were planted on a field site near a car factory in order to install a bioscreen. The aim was to combine the biodegradation activities of poplar and its associated rhizosphere and endophytic microorganisms for containing a BTEX contaminated groundwater plume. This BTEX plume occurred as the result of leaking solvents and fuel storage tanks. Monitoring, conducted overa 6-year period (1999-2005) after the planting of the trees suggested that the poplar trees and their associated microorganisms had, once the tree roots reached the contaminated groundwater zone, an active role in the remediation of the BTEX plume, resulting in full containment of the contamination. Analysis of the microbial communities associated with poplar demonstrated that, once the poplar roots got in contact with the BTEX contaminated groundwater, enrichment occurred of both rhizosphere and endophytic bacteria that were able to degrade toluene. Interestingly, once the BTEX plume was remediated, the numbers of toluene degrading rhizosphere and endophytic bacteria decreased below the detection limit, indicating that their population resulted from selective enrichment by the presence of the contaminants. PMID- 19810346 TI - Field note: irrigation of tree stands with groundwater containing 1,4-dioxane. AB - Coniferous and deciduous tree stands totaling 14 ha were recently planted on a closed landfill, and when mature, the stands are expected to be part of a natural treatment system for recovered groundwater. The trees would be irrigated at the rate of 189 L/min year-round with water containing 1,4-dioxane (< 10 mg/L), a compound that would be taken up and phytovolatilized by the trees. The water is moderately saline and contains elevated levels of manganese. This paper describes a concurrent series of preliminary studies, performed prior to the full-scale planting, to assess the feasibility of the phytoremediation system. Greenhouse experiments were carried out to identify tree species that can take up 1,4 dioxane and are tolerant of the water. Estimates were made of the area of the tree stand necessary to transpire the irrigation water plus precipitation. The landfill matrix was characterized in terms of its percolation rate and water holding capacity and based on those results salinity-modeling studies were carried out to estimate the fate and leaching potential of the various inorganic species that would accumulate in the root-zone of the trees. A pilot study, currently in progress on the landfill, suggested that the landfill cap is a suitable matrix for the establishment of large trees, and that the stands could be irrigated without the production of excess drainage. PMID- 19810347 TI - Technical note: nitrogen fertilization effects on the degradation of aged diesel oil in composted drilling wastes. AB - Hydrocarbon-contaminated wastes generated from oil and gas drilling activities may be used as a soil amendment once composted and further decomposition of residual hydrocarbons can be accomplished after the composts are applied to soils. To test if N fertilization may enhance hydrocarbon decomposition, we investigated the effects of N application on hydrocarbon degradation in different aged composts (1-, 2-, 3-, and 4-year-old composts, coded as 1Y, 2Y, 3Y, and 4Y composts, respectively) through a pot experiment planted with white spruce (Picea glauca [Moench] Voss) seedlings. The percentage degradation of total petroleum hydrocarbon (TPH, C11 to C40) in the composts without N fertilization was correlated to initial NH4+ concentrations (R = 0.99, P < 0.001). The percentage degradation of TPH was highest in the 3Y compost (41.1%) that had an initial level of 325.3 mg NH4+ -N kg(-1) and the lowest in the IY compost (9.3%) that had an initial level of 8.3 mg NH4+ -N kg(-1). The degradation of TPH was enhanced by Nfertilization in the 1Y (from 9.3 to 15.3%) and 4Y composts (from 14.3 to 22.6%) that had low initial NH4+ concentrations. Our results show that application of NH4+ -based fertilizers may enhance the degradation of TPH when initial NH4+ concentrations in the compost are low. PMID- 19810348 TI - Phytoremediation in education: textile dye teaching experiments. AB - Phytoremediation, the use of plants to clean up contaminated soil and water, has a wide range of applications and advantages, and can be extended to scientific education. Phytoremediation of textile dyes can be used as a scientific experiment or demonstration in teaching laboratories of middle school, high school and college students. In the experiments that we developed, students were involved in a hands-on activity where they were able to learn about phytoremediation concepts. Experiments were set up with 20-40 mg L(-1) dye solutions of different colors. Students can be involved in the set up process and may be involved in the experimental design. In its simplest forms, they use two week-old sunflower seedlings and place them into a test tube of known volume of dye solution. Color change and/or dye disappearance can be monitored by visual comparison or with a spectrophotometer. Intensity and extent of the lab work depends on student's educational level, and time constraints. Among the many dyes tested, Evan's Blue proved to be the most readily decolorized azo dye. Results could be observed within 1-2 hours. From our experience, dye phytoremediation experiments are suitable and easy to understand by both college and middle school students. These experiments help visual learners, as students compare the color of the dye solution before and after the plant application. In general, simple phytoremediation experiments of this kind can be introduced in many classes including biology, biochemistry and ecological engineering. This paper presents success stories of teaching phytoremediation to middle school and college students. PMID- 19810349 TI - Differences in PAH desorption and sediment organic matter composition between non vegetated and recently vegetated fuel-oiled sediments. AB - We assessed the desorption behavior of pyrene, chrysene, phenanthrene, and tri alkylated (C3) phenanthrene/anthracenes for non-vegetated and recently vegetated (< 2 yrs) fuel-oiled sediments collected from the Indiana Harbor Canal (IHC), Gary, IN. Bulk sediment and humin were analyzed for PAH concentrations, organic matter composition, and PAH desorption behavior. PAH desorption isotherms and kinetics were determined using batch aqueous extractions and a two compartment, first-order kinetic model Vegetated sediments contained more plant carbon and were more nonpolar and less oxidized than non-vegetated sediments. Desorption kinetics indicated that PAH desorption was primarily controlled by a slow PAH desorbing fraction (F2) of IHC sediments. However, in vegetated sediments, particularly humin, PAH release from a faster PAH-desorbing fraction (F1) increased as did the rates (k2) of PAH desorption from the dominant slow PAH desorbing fraction (F2). We propose that vegetation provides aliphatic, nonpolar carbon to IHC sediments that facilitates more rapid PAH desorption from bulk sediment and humin. PMID- 19810350 TI - Cd and Zn accumulation in plants from the Padaeng zinc mine area. AB - Significant cadmium (Cd) contamination In soil and rice has been discovered in Mae Sot, Tak province, Thailand where the rice-based agricultural systems are established in the vicinity of a zinc mine. The prolonged consumption of Cd contaminated rice has potential risks to public health and health impacts of Cd exposed populations in Mae Sot have been demonstrated. The Thai government has prohibited rice cultivation in the area as an effort to prevent further exposure. Phytoextraction, the use of plants to remove contaminants from soil, is a potential option to manage Cd-contaminated areas. However, successful phytoextraction depends on first identifying effective hyperaccumulator plants appropriate for local climatic conditions. Five sampling sites at Padaeng Zinc mine, Tak province were selected to collect plant and soil samples. Total Cd and Zn concentrations in sediments or soils were approximately 596 and 20,673 mg kg( 1) in tailing pond area, 543 and 20,272 mg kg(-1) in open pit area, 894 and 31,319 mg kg(-1) in stockpile area, 1458 and 57,012 mg kg(-1) in forest area and 64 and 2733 mg kg(-1) in Cd contaminated rice field. Among a total of 36 plant species from 16 families, four species (Chromolaena odoratum, Gynura pseudochina, Impatiens violaeflora and Justicia procumbens) could be considered as Cd hyperaccumulators since their shoot Cd concentrations exceeded 100 mg Cd kg(-1) dry mass and they showed a translocation factor >1. Only Justicia procumbens could be considered as a Zn hyperaccumulator (Zn concentration in its shoot more than 10,000 mg Zn kg(-1) dry mass with the translocation factor >1). PMID- 19810351 TI - Chloride and sodium uptake potential over an entire rotation of Populus irrigated with landfill leachate. AB - There is a need for information about the response of Populus genotypes to repeated application of high-salinity water and nutrient sources throughout an entire rotation. We have combined establishment biomass and uptake data with mid- and full-rotation growth data to project potential chloride (Cl-) and sodium (Na+) uptake for 2- to 11-year-old Populus in the north central United States. Our objectives were to identify potential levels of uptake as the trees developed and stages of plantation development that are conducive to variable application rates of high-salinity irrigation. The projected cumulative uptake of Cl- and Na+ during mid-rotation plantation development was stable 2 to 3 years after planting but increased steadily from year 3 to 6. Year six cumulative uptake ranged from 22 to 175 kg Cl- ha(-1) and 8 to 74 kg Na+ ha(-1), while annual uptake ranged from 8 to 54 kg Cl- ha(-1) yr(-1) and 3 to 23 kg Na+ ha(-1) yr(-1). Full-rotation uptake was greatest from 4 to 9 years (Cl-) and 4 to 8 years (Na+), with maximum levels of Cl- (32 kg ha(-1) yr(-1)) and Na+ (13 kg ha(-1) yr(-1)) occurring in year six. The relative uptake potential of Cl- and Na+ at peak accumulation (year six) was 2.7 times greater than at the end of the rotation. PMID- 19810352 TI - Phytoremediation of BTEX hydrocarbons: potential impacts of diurnal groundwater fluctuation on microbial degradation. AB - Volatile hydrocarbons have multiple potential fates in phytoremediation. This research investigated the relationship between biodegradation and plant uptake of BTEX compounds in laboratory and field settings. At a phytoremediation site, preliminary studies revealed minimal uptake into trees and enhanced degradation potential in the rhizosphere and in the bulk soiL Increased oxygen transport to the vadose zone caused by diurnal rise and fall of the water table was hypothesized to enhance degradation in the bulk soil. A detailed greenhouse study was then conducted to investigate potential bioremediation impacts using field site soil and DN34 hybrid poplar trees. In rhizosphere soils, the contaminated planted reactor had significantly higher BTEX degrader populations versus the uncontaminated-planted reactor, as was anticipated. The bulk soil in the planted contaminated reactor had increased degrader populations than the unplanted contaminated soil or planted-uncontaminated soil, and planting increased degradation throughout the soil profile, not just in the limited volume of rhizosphere soils. Oxygen diffusive and advective transport into reactors was modeled and calculated. Oxygen input in planted reactors was at least 3 to 5 times higher than in unplanted reactors, and increasing oxygen input lead to increased degrader populations in a linear manner. These results combined with the knowledge that high-transpiration trees draw the contaminated groundwater to the capillary fringe and the rhizosphere indicate that phytoremediation can aid microbial degradation via multiple mechanisms: increasing degrader populations, increasing oxygen input via groundwater diurnal fluctuations, and transporting contaminants to the biologically-enriched soil profile. PMID- 19810353 TI - Rhizosphere microbial densities and trace metal tolerance of the nickel hyperaccumulator Alyssum serpyllifolium subsp. lusitanicum. AB - In this study we determine culturable microbial densities (total heterotrophs, ammonifiers, amylolytics and cellulolytics) and bacterial resistance to Co, Cr, and Ni in bulk and rhizosphere soils of three populations of the Ni hyperaccumulator Alyssum serpyllifolium subsp. lusitanicum and the excluder Dactylis glomerata from ultramafic sites (two populations in Northeast (NE) Portugal (Samil (S), Morais (M)) and one population in Northwest (NW) Spain (Melide (L)). The relationship between bioavailable metal concentrations (H2O soluble) and microbial densities were analysed. Significant differences in microbial densities and metal-resistance were observed between the two species and their three populations. The hyperaccumulator showed higher microbial densities (except cellulolytics) and a greater rhizosphere effect, but this was only observed in S and M populations. These populations of A. serpyllifolium also showed selective enrichment of Ni-tolerant bacteria at the rhizosphere where Ni solubility was enhanced (densities of Ni-resistant bacteria were positively correlated with H2O-soluble Ni). These rhizobacteria could solubilise Ni in the soil and potentially improve phytoextraction strategies. PMID- 19810354 TI - Phytostabilization of amended soils polluted with trace elements using the Mediterranean shrub: Rosmarinus officinalis. AB - We evaluate the mid-term effects of two amendments and the establishment of R. officinalis on chemical and biochemical properties in a trace element contaminated soil by a mine spill and the possible use of this plant for stabilization purposes. The experiment was carried out using containers filled with trace element polluted soil, where four treatments were established: organic treatment (biosolid compost, OAR), inorganic treatment (sugar beet lime, IAR), control with plant (NAR) and control without plant (NA). Amendment addition and plant establishment contributed to restore soil chemical (pH, total organic carbon, and water soluble carbon) and biochemical properties (microbial biomass carbon and the enzymatic activities: aryl-sulphatase and protease). The presence of rosemary did not affect soluble (0.01 M CaCl2) Cd and Zn and decreased trace element EDTA extractability in amended soils. There were no negative effects found on plant growth and nutrient content on polluted soils (NAR, OAR, and IAR). Trace element contents were within normal levels in plants. Therefore, rosemary might be a reliable option for successful phytostablization of moderate trace element contaminated soils. PMID- 19810355 TI - Comparison of EDTA- and citric acid-enhanced phytoextraction of heavy metals in artificially metal contaminated soil by Typha angustifolia. AB - A pot experiment was conducted to study the performance of EDTA and citric acid (CA) addition in improving phytoextraction of Cd, Cu, Pb, and Cr from artificially contaminated soil by T. angustifolia. T. angustifolia showed the remarkable resistance to heavy metal toxicity with no visual toxic symptom including chlorosis and necrosis when exposed to metal stress. EDTA-addition significantly reduced plant height and biomass, compared with the control, and stunted plant growth, while 2.5 and 5 mM CA addition induced significant increases in root dry weight. EDTA, and 5 and 10 mM CA significantly increased shoot Cd, Pb, and Cr concentrations compared with the control, with EDTA being more effective. At final harvest, the highest shoot Cd, Cr, and Pb concentrations were recorded in the treatment of 5 mM EDTA addition, while maximal root Pb concentration was found at the 2.5 mM CA treatment. However, shoot Cd accumulation in the 10 mM CA treatment was 36.9% higher than that in 2.5 mM EDTA, and similar with that in 10 mM EDTA. Shoot Pb accumulation was lower in 10 mM CA than that in EDTA treatments. Further, root Cd, Cu, and Pb accumulation of CA treatments and shoot Cr accumulation in 5 or 10 mM CA treatments were markedly higher than that of control and EDTA treatments. The results also showed that EDTA dramatically increased the dissolution of Cu, Cr, Pb, and Cd in soil, while CA addition had less effect on water-soluble Cu, Cr, and Cd, and no effect on Pb levels. It is suggested that CA can be a good chelator candidate for T. angustifolia used for environmentally safe phytoextraction of Cd and Cr in soils. PMID- 19810356 TI - Phytoremediation of metal-contaminated soil in temperate humid regions of British Columbia, Canada. AB - The suitability of five plant species was studied for phytoextraction and phytostabilisation in a region with temperate maritime climate of coastal British Columbia, Canada. Pot experiments were conducted using Lolium perenne L (perennial rye grass), Festuca rubra L (creeping red fescue), Helianthus annuus L (sunflower), Poa pratensis L (Kentucky bluegrass) and Brassica napus L (rape) in soils treated with three different metal (Cu, Pb, Mn, and Zn) concentrations. The bio-metric characters of plants in soils with multiple-metal contaminations, their metal accumulation characteristics, translocation properties and metal removal were assessed at different stages of plant growth, 90 and 120 DAS (days after sowing). Lolium was found to be suitable for the phytostabilisation of Cu and Pb, Festuca for Mn and Poa for Zn. Metal removal was higher at 120 than at 90 days after sowing, and metals concentrated more in the underground tissues with less translocation to the aboveground parts. Bioconcentration factors indicate that Festuca had the highest accumulation for Cu, Helianthus for Pb and Zn and Poa for Mn. PMID- 19810357 TI - Effect of pH, temperature, and lead concentration on the bioremoval of lead from water using Lemna minor. AB - This study examined the ability of the aquatic plant Lemna minor (duckweed) to remove soluble lead under various laboratory conditions. In a batch process L. minor was exposed to different pH values (4.5-8.0) and temperature (15-35 degrees C) in presence of different lead concentrations (0.1-10.0 mg L(-1)) for 168 h. The amount of biomass obtained in the study period on a dry weight basis, the concentrations of lead in tissue and in medium and net uptake of lead by Lemna all have been determined in each condition. The percentages of lead uptake ratios (PMU) and bioconcentration factors (BCF) were also calculated for these conditions. Bioaccumulated lead concentrations and the PMU were obtained at lowest pH of 4.5, and at 30 degrees C. The highest accumulated lead concentration was found at pH 4.5 as 3.599 mg Pb g(-1) in 10.0 mg L(-1). It decreased to pH 6.0, but it did not change at pH 6.0-8.0 range. The maximum lead accumulation was obtained at 30 degrees C as 8.622 mg Pb g(-1) in 10 mg L(-1) at pH 5.0, and the minimum was at 15 degrees C as 0.291 mg g(-1) in 0.1 mg L(-1). Lead accumulation gradually increased with increasing lead in medium, but the opposite trend was observed for PMU. Lead accumulation increased up to 50 mg L(-1), but did not change significantly in the 50.0-100.0 mg L(-1) range. The lead uptake from water was modeled and the equation fit the experimental data very well PMID- 19810358 TI - Pyrene effects on rhizoplane bacterial communities. AB - Certain plant species promote biodegradation of polycyclic aromatic hydrocarbons (PAHs), but few studies have examined the microbial populations that are associated with the rhizoplane of these plants. In this study, the bacterial composition of the rhizoplane were characterized for four plant species during in soils with different histories of exposure to PAH and in the presence or absence of a pyrene spike at 100 mg kg(-1) pyrene. Three of the plant species including Andropogon gerrardii, Panicum coloratum and Melilotus officinalis were known to stimulate PAH degradation. Wheat (Triticum aestivum) was used as a reference species. Results showed that after 90 days, approximately 45% of the pyrene spike disappeared from soil without plants. In contrast, cultivation of plants resulted in 95% disappearance of pyrene. There were no significant differences in the extent of pyrene disappearance for different plants. In all cases, 16S rRNA gene profiles of the rhizoplane were less complex in the pyrene-spiked soils, suggesting that richness and evenness of the predominant bacteria were reduced. Our results show that pyrene contamination results in significant shifts in the composition of rhizosphere bacterial communities that are still further influenced by the plant species and prior exposure history to PAH contamination. PMID- 19810359 TI - Phylogenetic variation in the tolerance and uptake of organic contaminants. AB - An investigation into the phylogenetic variation of plant tolerance and the root and shoot uptake of organic contaminants was undertaken. The aim was to determine if particular families or genera were tolerant of or accumulated organic pollutants. Data were collected from sixty-nine studies. The variation between experiments was accounted for using a residual maximum likelihood analysis to approximate means for individual taxa. A nested ANOVA was subsequently used to determine differences at a number of differing phylogenetic levels. Significant differences were observed at a number of phylogenetic levels for the tolerance to TPH, the root concentration factor and the shoot concentration factor. There was no correlation between the uptake of organic pollutants and that of heavy metals. The data indicate that plant phylogeny is an important influence on both the plant tolerance and uptake of organic pollutants. If this study can be expanded, such information can be used when designing plantings for phytoremediation or risk reduction during the restoration of contaminated sites. PMID- 19810360 TI - Survival and growth of the dominant salt marsh grass Spartina alterniflora in an oil industry saline wastewater. AB - Saline oil produced water (PW) is the largest wastewater stream in the oil exploration and production processes. Although eventual disposal of PW into shallow coastal waters occurs nearby coastal wetlands, no studies regarding its toxicity to higher plants were found in our literature review. To fill this knowledge gap and evaluate the potential use of this halophyte for PW phytoremediation the salt marsh grass Spartina alterniflora was grown in five PW concentrations and no PW treatment control for seven weeks. The oil & grease, NaCl, and ammonium (N-NH4+) concentrations in the PW were 120 mg L(-1), 30 g L( 1), and 381 mg L(-1), respectively. Plants grown in 30% PW and 10% PW achieved survival rates (75%) significantly higher than plants grown in 100% PW (35% survival). LT50 of S. alterniflora to raw PW with 120 mg L(-1) of oil & grease (100% PW) was estimated at 30 days. Root and sprout biomass were significantly stimulated by PW; plants grown in 10% to 50% PW concentrations were 70-300% more productive than those in control, 80% PW and 100% PW, respectively. No significant inhibitory effects on survival or growth were detected for concentrations of PW less than 80% when compared to control. Our results pointed out that S. alterniflora grows in saline oil PW and its potential use to phytoremediate this effluent should be evaluated. PMID- 19810361 TI - Save lives: make hospitals safe for emergencies. PMID- 19810362 TI - Penicillium morneffei, an emerging AIDS-related pathogen--a RIMS study. AB - Twenty-seven HIV-infected patients with penicilliosis were included to study the clinical, laboratory and therapeutic features of HIV-infected patients with disseminated Penicillium marneffei infection. Diagnosis was based on microscopy and culture. CD4 counts were assessed by flow cytometry. Males (23 out of 27) predominated while the ages ranged from 20 to 54 years (mean 31.5 years). The most common manifestations were fever, weight loss, anaemia and skin lesions. CD4 counts were low in all the patients. The response to treatment with intravenous amphotericin B followed by itraconazole was excellent. Penicillium marneffei has become an important opportunistic pathogen of HIV infection in Manipur. Penicilliosis should be considered in a susceptible patient with a non-specific febrile illness, a low CD4 count and molluscum contagiosum-like skin lesions. Early diagnosis and treatment improve survival. PMID- 19810363 TI - Anthropometric failure, a new approach to measure undernutrition: an experience from a rural community of West Bengal, India. AB - Undernutrition is the major threat to the health and well-being of preschool children in India. To assess the prevalence of undernutrition along with the risk factors among 1-3 years children, a community-based cross-sectional study was undertaken in villages served by Matigara sub-centre in Darjeeling district of West Bengal, India. Anthropometric measurement of 256 children was done as per WHO guidelines along with collection of sociocultural and healthcare related information. Z-score was calculated (NCHS reference) and the prevalence of stunting, underweight, and wasting was 46.9%, 52.3% and 15.2% respectively. Composite, index of anthropometric failure was 65.6%. Anthropometric failure was significantly associated with low income, more number of siblings, nuclear family, short duration of exclusive breastfeeding, later initiation of complementary feeding, immunisation status, mothers' BMI, episodes of diarrhoea and acute respiratory infection. Multipronged intervention is to be initiated to address this staggering problem of undernutrition. PMID- 19810364 TI - Prevalence of epilepsy in school-going children (6-18 years) in Kashmir Valley of North-west India. AB - Epilepsy is a common neurological disorder which predominantly affects paediatric and young adolescent population. The aim of present study was to evaluate the prevalence of epilepsy in school-going children (6-18 years) in Kashmir valley. Forty-nine cases of epilepsy were detected after screening 15218 school-going children. It included 55.1% (prevalence of 3.74/1000) males and 44.9% (prevalence of 3.13/1000) females. Rural-urban specification revealed a prevalence of 3.49/1000 and 2.96/1000 respectively. Age-specific prevalence was found to be 3.82/1000 (6-10 years), 3.44/1000 (11-14 years) and 2.33/1000 (15-18 years). Highest prevalence of epilepsy was found in upper lower class (prevalence of 3.89/1000) and lowest in lower middle class (prevalence of 2.78/1000). Again, higher prevalence (3.38/1000) was observed in children from government run educational institutions. Generalised tonic-clonic seizures (73.5%) was the commonest type of seizure observed. PMID- 19810365 TI - Comparative evaluation of the efficacy of four topical medications individually or in combination to treat grade I acne vulgaris. AB - A randomised study was carried out to evaluate the efficacy of four topical medications individually and in combination to treat grade I acne vulgaris which is characterised by mild lesions (< 10 in one side of face) consisting of predominantly comedones with occasional pustules in oily skin. Maintaining the inclusion and exclusion criteria 100 patients were selected and divided into 5 groups to receive different topical drugs at random basis in the dermatology OPD. Topical medication given to them is mentioned below against each group: Group I- retinonic acid, group II--benzoyl peroxide, group III--clindamycin, group IV- cleanser and group V--all the four medications. The patients were observed for reduction in number of comedones, suppression of papulopustules with healing rate, effects on facial skin, and rate of recurrence. Results were observed according to the groups. In group I old acne was reduced in size and gradually cleared off (80%). Recurrence was few with appearance of new microcomedones which were cleared off within short time. Skin became smoother and fresh. Texture became lighter in colour. In group II whiteheads were reduced at about 70% in number. Rate of recurrence was normal. Skin became rough and dry. In group III pustular acne healed better and faster. In group IV acne of oily skin healed better and faster. Rate of recurrence was normal. Skin became fresh and oil-free. In group V reduction of lesions was very much significant (90%) with quick healing rate of the comedones. Recurrence was normal but delayed. Skin became smoother, finer and fresher. So, combination therapy is better. Cleanser is always helpful even without medications. PMID- 19810366 TI - Is total thyroidectomy justified in multinodular goitre. AB - Role of total thyroidectomy in treatment of patients with benign diseases of thyroid gland remains controversial. The hypothesis behind the study is that total thyroidectomy can be performed in all benign thyroid diseases with equal rate of immediate and late complications compared with those of subtotal thyroidectomy. The retrospective case control study was done by reviewing the case records of patients with benign thyroid diseases involving both lobes operated during May, 2002 to September, 2004. The minimum follow-up period was 3 years. Of the 189 patients included in the study 94 underwent total thyroidectomy and 95 patients underwent subtotal thyroidectomy. There was no incidence of permanent recurrent laryngeal nerve paralysis in both groups. Temporary unilateral recurrent laryngeal nerve paralysis occurred in 3 patients (3.2%) in total thyroidectomy group and 1 (1.1%) patient in subtotal thyroidectomy group, which was not statistically significant (p = 0.307). Temporary hypocalcaemia was noted in 10 patients of total thyroidectomy group (10.6%) and 3 patients of subtotal thyroidectomy group (3.2%). Permanent hypocalcaemia was noted in 2 patients each in total thyroidectomy group and subtotal thyroidectomy group (p = 0.991). Total thyroidectomy is an acceptable alternative to subtotal thyroidectomy in benign diseases, considering the higher rate of postoperative morbidity in recurrent goitres. PMID- 19810367 TI - Clinical significance of pyloric aperture in the aetiology of peptic ulcer disease: a prospective study. AB - Despite so much contributions reported in the literature, the aetiology of the duodenal ulcer remains an enigmatic subject to the medical profession. Findings of Helicobacter pylori seem to have overshadowed the real issue, in that, how a small area of the duodenal mucosa could be inflicted with the acid-pepsin injury has not been questioned? One hundred and sixty-eight consecutive patients, presented with epigastric pain were included in the endoscopic study. The aim of the study was to find out the prevalence and its clinical importance on the sizes of the pyloric aperture in the aetiology of peptic ulcer disease. Demographic data on the sizes of the pyloric aperture were divided into two groups, in that, those up to 3 mm in diameter were included in one and those over the size of 3 mm in another. Among the 168 cases, the gastric ulcer was found in 12 and duodenal ulcer in 27 patients. The sex ratio of men to women was 1.4:1 found in the former and 8:1 in the latter. Among other findings, a knuckle of duodenal mucoa was noticed prolapsing through the large pyloric aperture. It could be postulated that a knuckle of the mucosa that keeps peeping through the pylorus acts as a mucosal plug in empty stomach, like a cork in the acid bottle. The main physiological function is to protect the mucosa from being damaged by the acid pepsin injury or by the reflux of bile, but the tip of the plug seems to be subjected to such injury. Furthermore, the surface epithelial cells could also be subjected to ischaemic change while prolapsing through the pylorus. This may lead to reduced production of the mucosal gel and bicarbonate secretion, thus exposing the damaged mucosa to acid bath. This supports the concept, how a small area of the stomach or duodenum could be inflicted with ulceration. PMID- 19810368 TI - Successful pregnancy outcome among women with end-stage renal disease requiring haemodialysis. AB - Pregnancy is rare in women with end-stage renal disease, and perinatal outcome remains suboptimal because of prematurity and foetal growth restriction. Successful obstetrical outcome in two women presented with chronic renal failure requiring serial haemodialysis and multiple blood transfusions during pregnancy is reported. Both women had vaginal delivery of low birth weight neonates--2100 g and 1540 g at 33 and 37 weeks' gestations respectively. With specialised neonatal care, both neonates survived, and the mothers were counselled for renal replacement therapy. PMID- 19810369 TI - Unusual branching pattern of facial artery and a short review. AB - A detailed knowledge of the facial blood supply is necessary in planning of different types of orofacial surgeries. However abnormal pattern of facial blood supply may modify the result of the treatment, if it is not predicted in advance. Here we report two rare cases of unusual presentation of facial blood supply and the clinical implications of such findings, correlating with the previous studies. PMID- 19810370 TI - Right sided mammary hamartoma--a case report. AB - Mammary hamartoma is an uncommon benign lesion that presents as a palpable or impalpable mass in the breast. The lesion may be underdiagnosed by the pathologist, as the pathological appearance is not distinctive. Though mammary hamaroma is benign in nature, carcinomas can arise in it. Hence awareness of this lesion and a correct diagnosis is essential. A case of right sided mammary hamartoma clinically presenting as fibro-adenoma in a 38 years old female is reported. Lumpectomy was performed. Histopathological diagnosis confirmed it to be a case of mammary hamartoma. PMID- 19810371 TI - A rare presentation in a case of parathyroid carcinoma. AB - Parathyroid carcinoma is a very rare endocrine malignancy, which usually presents with features of hypercalcaemia and a neck mass. Here a case of parathyroid carcinoma, whose only presenting feature was multiple pathological fractures, without any palpable neck mass is reported. En masse resection of the parathyroid mass along with ipsilateral hemithyroidectomy was performed, while the fractures were treated conservatively. At three years follow-up the patient does not have any recurrence or any evidence of metastasis. PMID- 19810372 TI - Youngest patient with Sydenham's chorea: a case report. AB - Rheumatic fever is common in India and the incidence has not shown the declining trends seen in the developed countries. Two-thirds of children with rheumatic fever are school aged (5-15 years of age). However, rheumatic fever does occur in children < 5 years of age and it is believed that younger children are more likely to have carditis and arthritis and less likely to have chorea. A case of rheumatic chorea occurring in a 3 4/12-year-old boy is reported. The patient was treated with penicillin, prednisolone and sodium valproate and responded favourably. PMID- 19810373 TI - Malignant melanoma of vagina--a case report. AB - Malignant melanoma of vagina called mucosal lentigenous melanomas are extremely malignant. Malignant malanoma is a very rare tumour, it accounts for less than 3% of malignant tumours of vagina. Vaginal malanomas are aggressive and according to some authors they are less suitable for radical excision. Therefore wide local excision may be preferred. Multidisciplinary treatment modalities are required for more advanced lesions particularly in patients who have vaginal melanoma lesion more than 4 mm (AJCC stage IIB) in depth and high risk group. PMID- 19810374 TI - Tuwo cases of empyema necessitatis--experience and management. AB - Two patients were admitted in the paediatric ward with the complaints of fever, cough, loss of appetite and swelling of the chest wall. Both the patients were subjected to a thorough clinical examination with needle aspiration and routine investigations like haemogram, Mantoux test, chest x-ray, computerised tomography scanning of the thorax. Both the patients were diagnosed as cases of empyema necessitatis. The first case had a burst chest wall on the second day of admission and underwent chest intercostal drainage. Later on, thoracotomy, decortication and reconstruction of the chest wall was done. The second patient had got chest drainage and there was no burst chest wall. No thoracotomy was done. Both the patients were discharged after full recovery without antituberculosis therapy. PMID- 19810375 TI - A rare cutaneous manifestation of acute myeloid leukaemia. PMID- 19810376 TI - Medical education. PMID- 19810377 TI - "Safe surgery saves lives"--practice it and change the scenario. PMID- 19810378 TI - Intra-uterine insemination: pregnancy rate in relation to number, size of pre ovulatory follicles and day of insemination. AB - The objective of the study was to analyse the pregnancy rate in intra-uterine insemination (IUI) in relation to pre-ovulatory follicular number, size and day of insemination. A retrospective analysis of 216 completed IUI cycles was used in an attempt to identify significant variables predictive of treatment success. Couples with unexplained infertility and male factor infertility underwent IUI with or without ovarian stimulation. The mean number of IUI cycles per patient was 4.1, the overall pregnancy rate was 27.3% per patient, and the pregnancy rate per cycle was 6.9%. The pregnancy rate was 4.4% when one follicle was produced, whereas with more than two follicles, the rate increased to 21.2%. Hormonal stimulation using clomiphene citrate and/or human menopausal gonadotrophin/follicle stimulating hormone yielded a significant higher pregnancy rate compared to IUI in natural cycles (10.3% versus 3.3%). Although not statistically significant, the pregnancy rate decreased with advancing age of woman. The results suggest that IUI is a useful method of assisted conception in unexplained infertility and higher pregnancy rates can be achieved with good patient selection and ovarian stimulation. PMID- 19810379 TI - Coinfection by human immunodeficiency virus, hepatitis B virus and hepatitis C virus in injecting drug users. AB - Detection of hepatitis B surface antigen (HBsAg), anti hepatitis C virus (HCV) antibody and anti human immunodeficiency virus (HIV) antibody was carried out in serum samples from 250 injecting drug users (IDUs) from a deaddiction centre, Shalom, in Churachanpur district, Manipur during the period of August-October, 2002. One hundred and forty-nine (59.6%) IDUs were positive for HIV antibody, 226 (90.4%) were positive for anti HCV antibody and 27 (10.8%) were positive for HBsAg. There was coinfection of HIV, hepatitis B virus (HBV) and HCV in 15 (6%) of the IDUs. The coinfection of HBV and HCV was found in 12 cases (4.8%) and coinfection of HIV and HCV was found in 131 cases (52.4%). HIV and HCV were the highest among the coinfection of IDUs. The IDUs were in sexually active age group with a risk of infection to their sexual partner. PMID- 19810380 TI - Multidrug-resistant Gram-negative bacteria in postoperative infections. AB - Gram-negative bacteria are responsible in an important way in hospital-associated infections and their reported resistance to multiple antimicrobials is a matter of grave concern. In this study, Gram-negative bacteria isolated from clinical samples of postoperative infected patients were identified and their antibiotic susceptibility patterns determined. The prevalence of multidrug-resistant Gram negative bacteria was 9.45% and the highest prevalence was seen in GI surgery department. Escherichia coli and klebsiella were the most common bacterial isolates. Amongst the 14 antibiotics tested, the percentage sensitivity against the multidrug-resistant Gram-negative isolates was highest for amikacin, meropenem, piperacillin + tazobactam and cefoperazone + sulbactam. PMID- 19810381 TI - Patterns of alcohol consumption in medical students. AB - A prospective study was carried out among undergraduate medical students of three different medical colleges in North India. The aim of this study was to assess the consumption pattern of alcohol among medical students and correlate psychiatric disturbance and parental alcohol consumption with the patterns of alcohol use in them. Using an anonymous, self-administered questionnaire, we surveyed 168 subjects who were at various stages of their undergraduate medical career. Alcohol was the frequently used substance by all groups; 56.57% ever used alcohol and 41.46% showed patterns of problem drinking. Alcohol dependence was found in 6.09% of the students; 71.95% students started consuming alcohol after admission in the medical college. Almost one-third of respondents (37.50%) were found to be clinically depressed, anxious, or experiencing psychiatric disturbances. Such students had a greater frequency of alcohol consumption (p = 0.05). Also, a strong association between positive family history of alcohol use/abuse and use of alcohol among medical students was found (p = 0.001). Alcohol abuse amongst medical students should be taken more seriously because their own attitudes towards substances may influence their professional behaviour. PMID- 19810382 TI - Incidence and outcome of arthritis in meningococcal disease. AB - Meningococcal outbreaks are a major health concern in Delhi and adjoining regions. Besides acute pyogenic meningitis, meningococcal disease can also manifest as vasculitis, dermatitis and arthritis. To study the frequency, characteristics and long-term outcome of joint involvement in May 2005 meningococcal outbreak in New Delhi, 24 patients with proven meningococcal disease admitted to the hospital from May, 2005 through August, 2005 were studied for occurrence and outcome of joint involvement, and were followed up and evaluated for any complications. The frequency of arthritis was found to be 20%, which is much higher than reported. Diplococci could be identified in the joint aspirate of all 5 patients who developed arthritis. All patients had features of acute septic arthritis which healed without residual deformity following arthrotomy. A significant percentage of patients can still be expected to develop acute septic arthritis in an outbreak of meningococcal meningitis, and a high index of suspicion should be kept for the same. Prompt diagnosis and management will lead to healing without complications in most cases. PMID- 19810383 TI - Acute renal failure in intensive care unit. AB - A retrospective study was done to analyse the incidence and outcome of acute renal failure among all patients admitted in intensive care unit (ICU) of Apollo Hospital, Secunderabad during the 12-month period from March 2003 to February 2004. Of the total 1100 patients admitted in ICU during the period, 75 patients were diagnosed with acute renal failure considering the primary markers as the levels of nitrogenous compounds in blood, i.e., blood urea nitrogen and serum creatinine. Sepsis in 24 (32%), hypoperfusion in 46 (61.3%), nephrotoxic drugs in 3, contrast induced in 1 and 20 patients with surgical ailments were the factors responsible for causing acute renal failure in various combinations. Thirty-three patients required dialytic support. Of the 75 patients, 29 died and of those who survived 9 had partial recovery. The overall mortality in ICU was 8.1% as compared to mortality among acute renal failure in ICU setting at 38.7%. Sepsis, oliguria and hypotension and scores over 120 on the Apache III scoring system were indicators of poor outcome. More than one pathogenetic factor playing a role in the development of acute renal failure in the majority. Acute renal failure can be prevented in several cases or the severity reduced by appropriate measures. PMID- 19810384 TI - Clinicomicrobiological profile of women with vaginal discharge. AB - One hundred and ten patients presenting with vaginal discharge were investigated for microbiological spectrum of vaginitis and outcome of treatment. Laboratory documented disorder could be ascertained in 86 patients (78%). Bacterial vaginosis was the commonest aetiology (43.6%), followed by candidiasis (10%), trichomoniasis (9.1%), senile vaginitis (5.4%) and vaginitis of unknown origin (5.4%). Endocervicitis was seen in 13.6% patients and urinary tract infection in 1.8%. Complete relief of symptoms was seen in 77 patients (70%). PMID- 19810385 TI - Quality DOTS management and empowering tuberculosis patients. AB - Central Tuberculosis Division (CTD) has covered whole of India under DOTS. IMA is a proud partner of RNTCP which is managed by CTD. International Standards for Tuberculosis Care is expected from all healthcare providers. The basic principles of care is same worldwide. IMA GFATM RNTCP PPM is completing 2 years of its inception. Sensitisation programme and district training programmes has yielded DOTS/DMC centres in the target states. IMA is having 100% commitment for containing tuberculosis in India. There are International Standards for quality management in tuberculosis control, some of the Standards are elaborated in this write-up. In the Indian context, DOTS needs some innovations that is discussed in this article. PMID- 19810386 TI - Primary hydatidosis of female genital tract: a case report. AB - Hydatid cysts are known to occur in most organs especially in the endemic areas. The diagnosis of primary hydatid cyst in female genital tract is rare and difficult. A high degree of clinical suspicion should be there for doing pre operative investigations to exclude hydatid cyst of female pelvis. The patient presented with a lump in lower part of abdomen, which enlarged gradually over last 2 years. A provisional diagnosis of malignant ovarian cyst was made pre operatively. Total abdominal hysterectomy and bilateral salpingo-oophorectomy was done and whole of the specimen was sent for histopathological examination. On macroscopic examination, one cystic mass was present on the right side of uterus adherent to its anterior wall and second one was in the Douglas' pouch adherent to left adenexa and surrounding structures. The patient responded well to the surgical treatment followed by albendazole administration. PMID- 19810387 TI - Epidcermolytic hyperkeratosis: a case report. AB - Epidermolytic hyperkeratosis is a rare autosomal dominant genodermatosis that presents at birth as generalised erythema, blisters and erosions. In subsequent periods, erythema and blistering improves but patients go on to develop hyperkeratosis scalingthat is especially prominent along joint flexures, neck, hands and feet. The disease is caused by mutations in either keratin 1 or 10. Treatment options include topical emollients containing glycerin, lactic acid, urea and alpha-hydroxy acid and topical and systemic retinoids. Here a rare case in a 23 years old male is reported with epidermolytic hyperkeratosis and treated successfully with mixture of topical emollients with retinoid and systemic isotretinoin. PMID- 19810388 TI - Iatrogenic hypercalcaemia--a case report. AB - A case of iatrogenic hypercalcaemia in a 63-year-old female presenting with low back pain and tingling and numbness in both lower limbs off and on is presented here along with a brief discussion of literature. The purpose of this case reporting is to highlight how vitamins can be dangerous if prescribed inadvertently. PMID- 19810389 TI - Double Meckel's diverticulum. AB - Meckel's diverticulum is present in 2% of the population. Duplications of the alimentary tract are rare congenital malformations. The small bowel is the commonest site of alimentary tract duplication. They mimic other surgical diseases and may result in significant morbidity if left untreated. Pre-operative diagnosis of Meckel's diverticulum is difficult. A case of duplication of Meckel's diverticulum is reported here. PMID- 19810390 TI - Macrodystrophia lipomatosa. PMID- 19810391 TI - Subconjunctival cysticercosis. PMID- 19810392 TI - Angiotensin-converting enzyme inhibitors (ACEIs), not angiotensin receptor blockers (ARBs), are preferred and effective mode of therapy in high cardiovascular risk patients. AB - Blockade of the renin-angiotensin system (RAS) plays an important role in the prevention and correction of cardiovascular diseases. Agents that block the RAS such as angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin receptor blockers (ARBs) are major in this league. There have been numerous clinical trials looking at the use of ACEIs and ARBs in hypertension, heart failure (HF), and other special population who remain at high risk for cardiovascular and cardiometabolic abnormalities. Overall, ACEIs are the first line agents, recommended for high cardiovascular risk patients and are supported suitably by worldwide therapeutic guidelines including class IA recommendation from American College of Cardiology (ACC)/American Heart Association. These recommendations are based on, large body of clinical results which overall supports ACEIs in reducing mortality, MI, stroke, and new-onset congestive heart failure, and their unique cardioprotective benefits in patients with diabetes, independent of coexistent atherosclerosis and concomitant nephropathy. Although, theoretically, ARBs offer improved blockade of the RAS system than ACEIs, their relative effectiveness in the treatment of HF and other comorbid cardiovascular conditions remains controversial as evident from clinical trial and meta-analysis results which shows that ARBs are not as effective in reducing mortality, rate of hospitalisation, prevention of nephropathic progression, etc. The results from the latest ONTARGET 'non-inferiority' trial has further elucidated the fact that ARBs are no better than ACEIs at reducing fatal and non-fatal cardiovascular events including MI and CV death. Although theoretically, combination of ACEIs and ARBs is an attractive therapeutic option as none of them block RAS completely, but it may also open the gate for supplementary collection of adverse events as has been evidenced in recent trials. Although, there are no data at present to precisely suggest the efficacy differences between all available ACEIs, there are trials which support that ramipril, a long acting ACEI with good tissue penetration, potent long-lasting inhibition of ACE may not be applicable to other available ACEIs. Ramipril also specifically reduces major adverse coronary and cerebrovascular events in post MI patients when compared to other ACEIs or placebo. When clinicians are faced with the choice of using either an ACEI or an ARB in high-risk patients, they should be mindful of the unique differences between each class of medication, particularly with respect to MI and CV death, and also the range of indications, cost and individual convenience. PMID- 19810393 TI - 'For the times they are a-changin'. PMID- 19810394 TI - Mastication and swallowing: 2. control. AB - In recent years, it has become clear that the neural mechanisms controlling chewing and swallowing emanate from centres in the brainstem. However, these activities may be modulated by conscious processes and by feedback from peripheral nerves.This review relates this knowledge to clinical dentistry and, in particular, to the possible relationship between craniomandibular dysfunctions and impaired control of the masticatory system. CLINICAL RELEVANCE: Dentists should understand the neuromuscular control of the masticatory apparatus as this can be relevant to the general health of patients as well as to craniomandibular disorders involving the jaw muscles and joints. PMID- 19810395 TI - The current status of materials for posterior composite restorations: the advent of low shrink. AB - Polymerization contraction, and the stresses associated with this, have presented problems with resin composite materials, particularly when used to restore cavities in posterior teeth. This paper summarizes the problems associated with polymerization contraction and examines methods used to overcome this, in particular, by the use of materials which have reduced percentage contraction when compared with traditional materials. CLINICAL RELEVANCE: Use of a material with reduced polymerization contraction should lead to simpler restoration placement. PMID- 19810396 TI - The application of fluoride varnish in the prevention and control of dental caries. AB - The method of applying fluoride varnish is described and the evidence of its effectiveness in the prevention and control of caries is summarized.The application of fluoride varnish should be an integral part of caries preventive programmes. CLINICAL RELEVANCE: This paper describes how to apply fluoride varnish and reviews its effectiveness in preventing caries. PMID- 19810397 TI - Bisphosphonates and osteonecrosis of the jaw--current thoughts. AB - Bisphosphonates are increasingly being used in a number of bone conditions, including osteoporosis and metastatic cancer. As a consequence, the reported cases of bisphosphonate-related osteonecrosis of the jaw (BONJ) have increased. BONJ remains rare in patients being treated for osteoporosis, but is commoner in cancer patients where the bisphosphonate doses are much higher and mode of delivery is intravenous. Recently, much more is known about classification and mode of action of the bisphosphonates. The potency of the various nitrogen containing bisphosphonates is dependent on a number of factors including bone binding, zeta potential and inhibition of the enzyme farnesyl pyrophosphate synthase. There is current debate on why BONJ affects the jaw, particularly in relation to whether the jaw has high bone turnover or not. More is being learnt about the micro-organisms involved in the pathogenesis of BONJ and there is increasing evidence on the role of surgical treatment of this disabling condition. CLINICAL RELEVANCE: Dentists should be aware of the association of bisphosphonates and BONJ. This paper provides recent knowledge on bisphosphonate action, pathogenesis of BONJ and its treatment. PMID- 19810398 TI - Gingival veneers. AB - Gingival veneers have been in clinical use for over 50 years and are a predictable, inexpensive, non-invasive, effective and aesthetically pleasing way of replacing lost tissue. They can also be used as delivery vehicles for topical medicaments. This article describes their indications, contra-indications and construction. CLINICAL RELEVANCE: Dentists in primary and secondary care should be aware of the use and fabrication techniques of gingival veneers. PMID- 19810399 TI - Role of forensic odontology in the world's major mass disasters: facts and figures. AB - Forensic identification of the victims in mass disasters is essential, not only for humanitarian reasons, but also for civil or criminal investigative need. The number of victims identified with the help of forensic odontology in various mass disasters in the world has been analysed and discussed. The result depicts the necessity of keeping proper dental records at institutional and individual level. Thus the use of forensic odontology in a series of mass disasters has been explored. The most common aspect of forensic odontology that a general practitioner is likely to encounter is the supply of ante-mortem records to aid in human identification. CLINICAL RELEVANCE: The need for proper dental record keeping by general dental practitioners is highlighted by discussing the role of forensic odontology in some of the world's major mass disasters. PMID- 19810400 TI - An update on oral hygiene products and techniques. PMID- 19810401 TI - Technique of providing provisional restoration for a badly broken down tooth. PMID- 19810402 TI - Hot topics in special care dentistry. 5. Transplant patients. PMID- 19810403 TI - Physical signs for the general dental practitioner. Verruca vulgaris. PMID- 19810404 TI - Aspects of human disease. 37. Addison's disease (primary hypoadrenocorticism). PMID- 19810405 TI - Interaction of proteins with linear polyelectrolytes and spherical polyelectrolyte brushes in aqueous solution. AB - We review recent experiments on the interaction of proteins with anionic polyelectrolytes in aqueous solution. Data from the literature demonstrate that proteins can form soluble complexes with linear polyelectrolytes even on the "wrong side" of the isoelectric point, that is, for pH values above the isoelectric point of the proteins under which the polyelectrolytes and the proteins are like-charged. All data published so far demonstrate that this type of adsorption becomes weaker with increasing ionic strength. A much stronger interaction is found if the polyelectrolyte chains are grafted onto solid surfaces to form polyelectrolyte brushes. Here it has been shown that spherical polyelectrolyte brushes consisting of a core of ca. 100 nm diameter and long attached polyelectrolyte chains strongly adsorb proteins at low ionic strength ("polyelectrolyte-mediated protein adsorption"; PMPA). Virtually no adsorption takes place onto the spherical polyelectrolyte brushes at high ionic strength. A critical comparison of data obtained on free polyelectrolytes and on polyelectrolyte brushes shows that both phenomena can be traced back to patches of positive charge on the surface of the proteins. Moreover, we discuss the driving force of the PMPA-process in terms of the Donnan pressure inside the brush layer. Here we find a good correlation which demonstrates that release of counterions during the process of adsorption is the main driving force. PMID- 19810406 TI - Ab initio and AIM theoretical analysis of hydrogen-bond radius of HD (D = F, Cl, Br, CN, HO, HS and CCH) donors and some acceptors. AB - Recently, we defined 'hydrogen-bond radii' for various hydrogen-bond donors, DH where D = F, Cl, Br, CN, HO or CCH from an empirical analysis. It was shown that the A...H distances in B...HD complexes could be written as a sum of hydrogen bond radius for DH and a constant acceptor radius for A, which is the bonding atom/centre in B. This manuscript reports the determination of the hydrogen-bond radii for these molecules and H2S from ab initio and atoms in molecules (AIM) theoretical calculations. The results from ab initio calculations are consistent with the empirical estimates for the six molecules noted above and provide the first estimate for hydrogen bond radius (1.08 +/- 0.16 A) for H2S donor. The results from AIM theoretical analysis are in qualitative agreement with ab initio results. However, AIM analysis indicates that both hydrogen bond donor and acceptor radii vary in a systematic way from the strong to weak hydrogen bonds. Irrespective of the method used, the hydrogen bond donor radius increases in the order HF < HCl < H2O < HBr < HCN < HCCH < H2S, but mostly lie between Pauling's covalent and van der Waals radii of H atom. Interestingly, the acceptor radii for A in A...HD also increase in the same order. The AIM theoretical results on about 100 complexes have been reduced to suggest radii for H, F, O, N, C and S that are appropriate for strong, medium and weak hydrogen bonds. It is suggested that the use of a single van der Waals radius for D, H or A in determining the presence/absence of D-H...A hydrogen bonding be discontinued. PMID- 19810407 TI - Density functional theory including dispersion corrections for intermolecular interactions in a large benchmark set of biologically relevant molecules. AB - Density functional theory including dispersion corrections (DFT-D) is applied to calculate intermolecular interaction energies in an extensive benchmark set consisting mainly of DNA base pairs and amino acid pairs, for which CCSD(T) complete basis set limit estimates are available (JSCH-2005 database). The three generalized gradient approximation (GGA) density functionals B-LYP, PBE and the new B97-D are tested together with the popular hybrid functional B3-LYP. The DFT D interaction energies deviate on average by less than 1 kcal mol(-1) or 10% from the reference values. In only six out of 161 cases, the deviation exceeds 2 kcal mol(-1). With one exception, the few larger deviations occur for non-equilibrium structures extracted from experimental geometries. The largest absolute deviations are observed for pairs of oppositely charged amino acids which are, however, not significant on a relative basis due to the huge interaction energies > 100 kcal mol(-1) involved. The counterpoise (CP) correction for the basis set superposition error with the applied triple-zeta AO basis sets varies between 0 and -1 kcal mol(-1) (<5% of the interaction energy in most cases) except for four complexes, where it is up to -1.4 kcal mol(-1). It is thus suggested to skip the laborious calculation of the CP correction in DFT-D treatments with reasonable basis sets. The three dispersion corrected GGAs considered differ mainly for the interactions of the hydrogen-bonded DNA base pairs, which are systematically too small by 0.6 kcal mol(-1) in case of B97-D, while for PBE-D they are too high by 1.5 kcal mol(-1), and for B-LYP-D by 0.5 kcal mol(-1). The all in all excellent results that have been obtained at affordable computational costs suggest the DFT D method to be a routine tool for many applications in organic chemistry or biochemistry. PMID- 19810408 TI - Photoinduced electron transfer sensitization investigated by chemically induced dynamic nuclear polarizatioin (CIDNP). AB - The key step of photoinduced electron transfer sensitization (sensitizer, S; donor substrate, M; quencher, Q) is the transformation of a primarily formed auxiliary radical ion pair M(*+)S(*-) into the desired radical ion pair M(*+)Q(* ). The systems chosen exhibit striking CIDNP effects, which can only be explained by that pair substitution. An inversion of all polarization phases occurs when the quencher diethylfumarate is added to the two-component system 9 cyanophenanthrene/ trans-anethole (S/M) because the g-value differences have opposite signs for the first and second radical pair. With the system 9 cyanoanthracene/trans-anethole/fumarodinitrile (S/M/Q), the pair substitution is a necessary condition for observable CIDNP, and no polarizations can be detected when any of the three components is omitted. Closed-form expressions for the CIDNP intensities as functions of the first-order rate constant of pair substitution k are derived, which use the limiting polarizations from the first and the second radical pair to eliminate any explicit dependence on diffusion related quantities and hyperfine coupling constants, leaving only k and the g value differences of the two pairs as parameters. From fits of these expressions to the dependence of the polarizations on the concentration of Q, the unknown g values of M(*+) and S(*-) are estimated. PMID- 19810409 TI - Exploration of the potential energy surface of C4H4 for rearrangement and decomposition reactions of vinylacetylene: a computational study. Part I. AB - The potential energy surface (PES) of C4H4 was explored using quantum chemical methods (DFT, MP2, MP4, GVB-MP2, CCSD(T), G2M, CBSQ/APNO) and 43 different structures located at global and local minima were identified. The majority of these structures correspond to carbenes, a minority to closed shell systems and biradicals (carbyne structures were not investigated). Whereas the chemistry of the closed shell systems such as vinylacetylene (1), butatriene (2), methylenecyclopropene (3), cyclobutadiene (5) or tetrahedrane (15) is well known, the carbenes represent unusual structural entities. 2-Methyl-cycloprop-2-en-1 ylidene (4) (DeltaDeltaH(298) = 36.2 kcal mol(-1) relative to 1) in its sigma2pi0 electron configuration at the carbene C of the 1A ground state is of comparable stability to cyclobutadiene (5) (DeltaDeltaH(298) = 33.4 kcal mol(-1); exp. value: 32.1 kcal mol(-1) as a result of aromatic 2pi-delocalization; carbene 3 vinylidenecyclopropene (13) (DeltaDeltaH(298) = 53.9 kcal mol(-1) does not possess C(2v) symmetry but has the vinylidene group bent toward the three membered ring (C(s)-symmetry) thus representing a frozen path point of the chelotropic addition of :C=C: to ethene. Allenyl carbene (14) has a triplet ground state and two low lying excited singlet states of closed shell (2.5 kcal mol(-1) higher) and open shell character (14.1 kcal mol(-1)). Carbene 14 is a crossing point on the C4H4 PES connecting closed-shell systems with each other. Because of the stability of 1, its rearrangement reactions are all connected with high activation enthalpies requiring 66 up to 92 kcal mol(-1) so that they energetically overlap with the activation enthalpies typical of decomposition reactions (from 90 kcal mol(-1) upward). The possible rearrangement reactions of 1 are investigated with a view to their relevance for the chemical behavior of the molecule under the conditions of Titan's atmosphere. PMID- 19810410 TI - Experimental and computational study of the ultraviolet photolysis of vinylacetylene. Part II. AB - The ultraviolet photochemistry of vinylacetylene (C4H4) was studied under temperature and pressure conditions similar to Titan's atmosphere by exciting the molecule in a constrained expansion that opens into the ion source region of a time-of-flight mass spectrometer. The primary dissociation products detected by vacuum-ultraviolet ionization were found to be C4H3 and C4H2, in a ratio of 3-10 : 1. Subsequent reaction of the C4H3 radicals with the parent C4H4 produced two major secondary products: C8H6 and C6H4. The former was spectroscopically identified as phenylacetylene, confirming that photochemical reactions of C4H4 can produce aromatic molecules. The primary dissociation reaction was also studied computationally. The results were consistent with the experimental findings for C4H2 and C4H3. However, the major product is C2H2, which is undetected by 118 nm photoionization in the present experiment but should account for roughly two-thirds of the products. Simulations were also performed to confirm that the present experiment accurately represents the 220 nm photochemistry of vinylacetylene at the temperature and pressure of Titan's atmosphere, with a product yield of C2H2 : C4H2 : C4H3 of 66 : 7 : 27. Accounting for the wavelength dependent solar flux on Titan, the estimated absorption cross section of vinylacetylene in the ultraviolet, and the slightly wavelength dependent product distribution, the overall product yield predicted by the simulations for ultraviolet photolysis of vinylacetylene on Titan is C2H2 : C4H2 : C4H3 = 65 : 8 : 27. Finally, a simulation was performed under conditions of a shock tube experiment to examine the differences between thermal and photochemical dissociation. The product yield of this simulation was C2H2 : C4H2: C4H3 = 61 : 1 : 38. PMID- 19810411 TI - Branching ratios for quenching of nitric oxide A 2Sigma+ (nu' = 0) to X 2Pi(nu' = 0). AB - We describe experiments designed to measure the fraction of nitric oxide molecules that undergo quenching from A 2Sigma+ (nu' = 0) directly to X 2Pi(nu" = 0). This quenching channel was investigated for room temperature collisions with O2, CO, CO2, and H2O by measuring recovery of the ground-state population following intense laser excitation. Experiments were conducted in a room temperature flow cell containing dilute mixtures of NO, N2, and the quenching gases. An intense nanosecond laser pulse, tuned to the NO A 2Sigma(+) - X 2Pi(0,0) Q11 + pQ21 bandhead at 226.3 nm, depopulated more than 20% of the equilibrium population in the X 2Pi(nu'' = 0) manifold. A weak, time-delayed, picosecond laser pulse, tuned to the A 2Sigma(+) - 2Pi(1,0) Q11 + pQ21 bandhead at 214.9 nm, probed recovery of population in X(nu'' = 0) via subsequent LIF for each of the investigated quenchers. Remarkably large branching ratios were observed for direct quenching to X 2Pi(nu'' = 0). Water, carbon monoxide, and oxygen quench NO A 2Sigma+ (v' = 0) to X 2Pi(nu" = 0) with branching ratios that are approximately 0.3. The significantly higher branching ratio for quenching by carbon dioxide is 0.6. The results provide insight on the NO quenching process and represent an important step toward a detailed understanding of the effects of collisional energy transfer on saturated laser-induced fluorescence, which is necessary to properly model detection strategies based on high laser fluences. PMID- 19810412 TI - Stepwise conformational cooling towards a single isomeric state in the four internal rotors system 1,2-butanediol. AB - The present work explores the possibilities of the matrix isolation technique in the structural characterisation of highly flexible molecules. To date, most studies of this type were carried out on molecules with three or less internal degrees of freedom and a few (less than 10) possible conformations. The molecule of 1,2-butanediol has four conformationally relevant three-fold rotational axes, which can result in 81 possible conformations. A detailed theoretical study, at the MP2 and DFT(B3LYP) levels of theory with the 6-311 + + G(d,p) basis set, revealed that more than 20 conformers of 1,2-butanediol have relative energies in a 0-10 kJ mol(-1) range and contribute appreciably to the gas phase equilibrium at room temperature. This fact renders conformational studies of the system extremely difficult under normal conditions. However, the method of matrix isolation permits the reduction of the number of populated conformational states in the experiment at low temperature due to the effect known as conformational cooling: low energy barriers promote the relaxation of the higher energy local minima into more stable structures. As a result of massive conformational cooling occurring upon matrix deposition, only five conformers of 1,2-butanediol were retained in the samples at 10 K. These conformers were identified using a combination of FTIR spectroscopy and extensive theoretical calculations of vibrational spectra. Annealing of the matrices up to 50 K resulted in the extreme case of conformational cooling related with the depopulation of all conformers into the most stable unique structure. The observed transformations were rationalized in terms of barriers to intramolecular rotation. PMID- 19810413 TI - The p-type conduction mechanism in Cu2O: a first principles study. AB - Materials based on Cu2O are potential p-type transparent semiconducting oxides. Developing an understanding of the mechanism leading to p-type behaviour is important. An accepted origin is the formation of Cu vacancies. However, the way in which this mechanism leads to p-type properties needs to be investigated. This paper presents a first principles analysis of the origin of p-type semiconducting behaviour in Cu2O with 1.5 and 3% Cu vacancy concentrations. Plane wave density functional theory (DFT) with the Perdew-Burke Ernzerhof (PBE) exchange correlation functional is applied. In order to investigate the applicability of DFT, we firstly show that CuO, with 50% Cu vacancies cannot be described with DFT and in order to obtain a consistent description of CuO, the DFT + U approach is applied. The resulting electronic structure is consistent with experiment, with a spin moment of 0.64 mu(B) and an indirect band gap of 1.48 eV for U = 7 eV. However, for a 3% Cu vacancy concentration in Cu2O, the DFT and DFT + U descriptions of Cu vacancies are similar, indicating that DFT is suitable for a small concentration of Cu vacancies; the formation energy of a Cu vacancy is no larger than 1.7 eV. Formation of Cu vacancies produces delocalised hole states with hole effective masses consistent with the semiconducting nature of Cu2O. These results demonstrate that the p-type semiconducting properties observed for Cu2O are explained by a small concentration of Cu vacancies. PMID- 19810414 TI - Redox competition mode of scanning electrochemical microscopy (RC-SECM) for visualisation of local catalytic activity. AB - In order to locally analyse catalytic activity on modified surfaces a transient redox competition mode of scanning electrochemical microscopy (SECM) has been developed. In a bi-potentiostatic experiment the SECM tip competes with the sample for the very same analyte. This leads to a current decrease at the SECM tip, if it is positioned in close proximity to an active catalyst site on the surface. Specifically, local catalytic activity of a Pt-catalyst modified sample with respect to the catalytic reduction of molecular oxygen was investigated. At higher local catalytic activity the local 02 partial pressure within the gap between accurately positioned SECM tip and sample is depleted, leading to a noticeable tip current decrease over active sites. A flexible software module has been implemented into the SECM to adapt the competition conditions by proper definition of tip and sample potentials. A potential pulse profile enables the localised electrochemically induced generation of molecular oxygen prior to the competition detection. The current decay curves are recorded over the entire duration of the applied reduction pulse. Hence, a time resolved processing of the acquired current values provides movies of the local oxygen concentration against x,y-position. The SECM redox competition mode was verified with a macroscopic Pt disk electrode as a test sample to demonstrate the feasibility of the approach. Moreover, highly dispersed electro-deposited spots of gold and platinum on glassy carbon were visualised using the redox competition mode of SECM. Catalyst spots of different nature as well as activity inhomogeneities within one spot caused by local variations in Pt-loading were visualised successfully. PMID- 19810415 TI - Comment on "Molecular dynamics DFT:B3LYP study of guanosinetriphosphate conversion into guanosinemonophosphate upon Mg2+ chelation of alpha and beta phosphate oxygens of the triphosphate tail" by Alexander A. Tulub, Phys. Chem. Chem. Phys., 2006, 8, 2187. AB - A recent paper in this journal uses molecular dynamics methods to study hydrolysis of guanosine triphosphate (GTP). The author reports that cleavage of the molecule occurs in less than 5 ps, and leads to a number of fragments including a free oxygen atom and a reduced magnesium ion. This conclusion is not in agreement with the known biochemistry and chemical reactivity of GTP or with previous computational studies of its hydrolysis reaction. PMID- 19810416 TI - Anaemia in the elderly. AB - Anaemia is highly prevalent in elderly populations, particularly in long term care facilities and geriatric wards. Even mild anaemia is associated with adverse health outcomes. Although senescence is considered to be a contributing factor, underlying pathology always has to be thoroughly explored. The most frequent etiologies of anaemia in the elderly are anaemia of chronic disease/inflammation; iron, folate and cobalamin deficiency; and myelodysplastic syndrome. Multiple concomitant etiologies are frequently present. The diagnostic workup is straightforward, not very invasive and should be done systematically. Nutrient deficiencies require complete workup and cure. Anaemia of chronic disease/inflammation ideally is taken care of by treating the underlying disease. If this is not possible and if glomerular filtration rate is significantly decreased, treatment with erythropoietin should be considered. Most cases of myelodysplastic syndrome will benefit from supportive care, but a specific subgroup responds particularly well to oral thalidomide analogues. Transfusions should be avoided. PMID- 19810417 TI - Sarcopenia and functional decline: pathophysiology, prevention and therapy. AB - Twenty years ago, the term 'sarcopenia' has been introduced to describe the ageing related loss of skeletal muscle mass. Since then, sarcopenia has been intensively studied and prevalence values have been reported in fifteen papers covering several continents and races. However, consistency regarding the outcome measures and corresponding cut-off values defining sarcopenia is lacking. Most approaches are based on estimations of muscle mass and proposed cut-off values might be too strict, thus reducing their use in daily practice. From a clinical viewpoint, the assessment of muscle performance (grip strength and endurance) can be proposed as a screening tool showing sufficient sensitivity. The pathophysiology of sarcopenia is multifactorial, and important changes at the tissue level have been identified. Close relationships with inflammatory processes have been demonstrated and there is strong evidence for the involvement of a chronic low-grade inflammatory activity. Sarcopenia is aggravated by a complex interaction of several factors among which aging, disuse, immobilization, disease and malnutrition. A comprehensive geriatric assessment should allow the clinician to estimate the relative contribution of these factors and to elaborate appropriate management. From all interventions studied, intensive resistance training seems the most efficient to counter sarcopenia, even in the very old geriatric patients. Significant ameliorations (up to >50% strength gain) can be expected after six weeks of training at a rhythm of 2-3 sessions per week. From a preventive viewpoint, all elderly patients should be advised to start such an exercise program and continue it as long as possible. To date, most pharmacological interventions to counter sarcopenia include drugs with anabolic effects. Unfortunately, their effect is questionable and no clear guidelines exist for the prescription of these products in the context of sarcopenia. PMID- 19810418 TI - Detection and identification of antinuclear antibodies (ANA) in a large community hospital. AB - OBJECTIVE: Antinuclear antibodies (ANA) and their identification are important diagnostic tools in rheumatic diseases. We aimed to determine their prevalence in samples referred for ANA testing and to identify factors predicting more specific reactivities. METHODS: We analyzed the first sample of 6422 consecutive patients for ANA. Positive samples were analyzed by indirect immunofluorescence (IIF) on Crithidia luciliae and by line immunoassay. We used multivariate logistic regression to detect predicting variables. RESULTS: 42.6% of all patients were ANA positive of which 13.0% showed > or = one extractable nuclear antigen (ENA) reactivity with anti-SSA/Ro (5.5%), anti-SSB/La (2.9%), anti-Cenp-B (2.5%) and anti-histones (2.2%) as the most prevalent antibodies. Anti-double-stranded DNA antibodies (dsDNA) were present in 1.0%. The strongest overall predictor was ANA intensity regardless of pattern. Cenp-B however was best predicted by pattern. Anti-dsDNA and anti-histone were more frequent in samples with a homogenous as compared with a speckled pattern. Anti-SSA and anti-SSB were more frequent in females and anti-Sm in patients < or = 30 years. CONCLUSIONS: The best overall predictor of antibodies to ENA or dsDNA is ANA intensity. Anti-Cenp-B is however best predicted by pattern. Samples with low ANA intensity (1+) may not need further testing unless a high clinical suspicion of ANA-associated disease. PMID- 19810419 TI - Influenza outbreak in a well-vaccinated nursing home population in Belgium. AB - Elderly people in nursing home communities are vulnerable to contagious infections, including the influenza virus. Systematic anti-influenza vaccination is an important preventive measure; however, vaccination does not provide absolute protection. We report an outbreak of influenza A infection in a well vaccinated nursing home population. Several factors can facilitate the occurrence of this type of outbreak. This report mainly addresses the discrepancy between the circulating viral strain and strains present in the recommended vaccine. PMID- 19810420 TI - CPAP treatment in the coexistence of obstructive sleep apnea syndrome and metabolic syndrome, results of one year follow up. AB - OBJECTIVE: To investigate the effect of one year continuous positive airway pressure (CPAP) treatment on metabolic syndrome (MS) prevalence and components in patients diagnosed with both obstructive sleep apnea syndrome (OSAS) and metabolic syndrome. METHODS: This was a single center, observational prospective cohort study. 38 patients who were diagnosed with OSAS after polysomnographic analysis in sleep laboratory and diagnosed with MS according to National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) guideline and underwent CPAP treatment were followed for one year. After the 1 year of follow up period on CPAP treatment, the prevalence of MS was evaluated again. RESULTS: 20 (13 male, 7 female) of 38 patients completed the entire study. Mean age was 50+/-7.7.4 patients were under treatment for diabetes mellitus (DM), 9 for hypertension (HT). After one year of follow up on CPAP treatment, the prevalence of MS decreased by 45%. When each components of MS were evaluated, no significant difference was found in fasting blood glucose, triglyceride levels and systolic and diastolic blood pressure after treatment (p>0.05). However, significant difference was observed in waist circumference (p=0.002), HDL cholesterol (p=0.001) and BMI (p=0.01) after treatment. DISCUSSION: If MS accompanies OSAS, which is a cardiovascular risk factor by itself, treatment indications of CPAP should be reevaluated. Thus, if OSA patients meet the criteria of MS even though they do not have obvious DM, HT and hyperlipidemia, initiating CPAP treatment at lower AHI levels may contribute to the prevention and development of cardiovascular disease. PMID- 19810421 TI - Urinary tract infections in spinal cord injury: prevention and treatment guidelines. AB - OBJECTIVES AND METHODS: The literature on prevention and therapy of urinary tract infection (UTI) in patients with spinal cord injury (SCI) was reviewed using 3 levels of evidence. RESULTS: Antibiotic therapy is only indicated in symptomatic bacteriuria or in symptomatic exacerbations of chronic UTI. During the acute phase of a SCI, UTI's are more prevalent and bacteria are different and more resistant to antibiotics compared with the chronic phase of SCI. In SCI in general, routine screening urine cultures are not valuable as a high species turn over is seen. Intermittent catheterisation, tapping or Crede manoeuvre coincide significantly with lower frequency of UTI compared to permanent catheter drainage. No measures are proven efficient in the long term in prevention of bacteriuria or UTI. Methenamine salts are perhaps useful in the prevention of UTI but not in patients with a permanent catheter (level III). Antibiotic prophylaxis was found useful in reducing asymptomatic bacteriuria but not in the prevention of symptomatic infections (level I). However, during prophylaxis a doubling of antibiotic resistance was found. In patients with augmented bladder antibiotic prophylaxis is useless (level II). In chronic SCI the first choice antibiotics are nitrofurantoin or trimethoprim, the second choice are fluoroquinolones (level III) whereas in acute SCI a higher resistance profile to antibiotics is frequent and therefore fluoroquinolones or cefuroxime are suggested (level III). There is no consensus in the literature but we suggest 5 days of antibiotic treatment in UTI during chronic SCI without fever, 7 days in acute SCI without fever and a minimum of 14 days in patients with UTI and fever (level III). PMID- 19810422 TI - Plasma vitamin C for predicting cardiovascular disease: more than a nutritional biomarker. PMID- 19810423 TI - General internal medicine in Belgium: to be or not to be? PMID- 19810424 TI - Campylobacter fetus cellulitis in an immunocompromised patient: case report and review of the literature. AB - Campylobacter fetus is an opportunist Gram-negative bacillus. The most frequent clinical manifestation is bacteriemia but it can also be responsable for soft tissue infections, endovascular infections, meningitis, peritonitis and thrombophlebitis. Campylobacter fetus cellulitis has been described, but rarely identified in subcutaneous puncture samples. We report a case of an immunocompromised patient with Campylobacter fetus bacteriemia associated with a soft tissue infection whose subcutaneous puncture also revealed the bacteria. PMID- 19810425 TI - Minocycline induced eosinophilic pneumonia: case report and review of literature. AB - A 51-year-old woman with eosinophilic pneumonia due to minocycline is described and a review of available literature is added. Until now, only 49 cases have been described, mainly in the Japanese population. Minocycline induced eosinophilic pneumonia is probably underreported and even underdiagnosed. This case highlights the importance of careful history taking, especially the use of drugs. Relatively safe drugs (like minocycline) can cause serious adverse events. On presentation, the disease mimics an infectious pneumonia. Peripheral eosinophilia can occur but isn't obligatory. A bronchoalveolar lavage may provide the first (and sometimes only) sign of eosinophilic lung disease. Withdrawal of minocycline is often enough although sometimes corticosteroids are needed. In general, prognosis is good when the diagnosis is made on time. PMID- 19810426 TI - Common variable immunodeficiency (CVID): case report and review of the literature. AB - Common variable immunodeficiency (CVID) is an immunodeficiency disease characterized by diminished ability to produce immunoglobulins. CVID has an estimated incidence of 1:10,000 to 1:200,000 (male:female 1:1) and usually presents in the second and third decade, although it also has a peak of incidence in childhood. The exact pathophysiology remains unclear. CVID can be associated with autoimmune, granulomatous and gastrointestinal diseases and patients have a predisposition to malignancies (especially non-Hodgkin lymphoma). Since different organ systems can be affected, all clinicians need to be aware of this entity, especially when confronted with patients with recurrent infections and/or multiple autoimmune diseases. PMID- 19810427 TI - Group C streptococcal psoas abscess associated with a homolateral hip joint prosthesis infection: a case report. AB - An abscess in the psoas muscle is rare and frequently misdiagnosed. A delay in the diagnosis can increase its mortality rate. Some clinical signs can help the clinician but they all are not always present, and not at the same time. We describe in this paper a case report of an association between a psoas abscess and a homolateral hip joint prosthesis infection. It was suspected because of no improvement in clinical state despite treatment of the abscess by antibiotics and drainage, and it required finally other complementary therapeutic solutions. The pathogenic microorganism was a group C streptococcus. We discuss all these points and thereafter we suggest some recommendations for the clinician. PMID- 19810428 TI - A not so healthy muesli: a case report. AB - We present a patient with severe anaphylaxis, angioedema, hypotension and shock. The near fatal allergic reaction was caused by eating yogurt with muesli containing pine nuts. The patient developed an acute infero-posterolateral myocardial infarction due to systemic hypotension and shock. Food allergy to pine nut was demonstrated by dosage of specific IgE to pine nut. PMID- 19810429 TI - Memes and suicide. AB - The concept of memes is analyzed, and its applicability to suicidology explored. Proposals are made for possible memes implicated in suicidal behavior. A classification of suicidal memes is proposed and the relationship between memes and archetypes of suicide is discussed. It is suggested that the terminology of meme theory can sharpen research into imitation effects in suicide. PMID- 19810430 TI - Heuristic versus systematic processing of information in detecting deception: questioning the truth bias. AB - Research on nonverbal detection of deception has normally been conducted by asking observers to judge the veracity of a number of videotaped communications. These video clips have typically been very short. Observers have a tendency to judge most of these statements as truthful. An experiment was conducted in which 52 participants (44 women, 8 men; M age = 22.2 yr., SD = 2.2) who were taking a psychology and law course were requested to make judgments of credibility at different points of the senders' statements. A strong truth bias was apparent when judgments were made at the beginning of the statements, suggesting that when exposed to brief communications, the observers make heuristic judgments. Over time, a decrease in the truth bias and an increase in overall accuracy were found, suggesting that later judgments were increasingly based on systematic information processing. These results suggest that the truth bias that has been found in previous deception research may be a result of having used very brief and uninformative behavioral samples as stimuli. PMID- 19810431 TI - Age-related change in emotional experience in a sample of Chinese adults: a preliminary study. AB - Age-related change in emotional experience was explored in a Chinese community sample. 968 healthy adults (483 women, 485 men) ranging in age from 18 to 66 years (M = 37.5, SD = 12.3) took part. The frequency of experiencing negative and positive emotion was self-reported on the general dimension scales of Positive and Negative Affect Scale-Expanded Form (PANAS-X). Regression analyses indicated that the frequency of negative affect decreased with age, and the frequency of positive affect was not significantly associated with age. These findings are similar to those found in Western samples, as discussed in relation to socioemotional selectivity theory. PMID- 19810432 TI - Psychometric properties of the Chinese translation of the proactive personality scale. AB - The purpose of the present study was to examine the psychometric properties of the Chinese translation of the Proactive Personality Scale. Four samples were surveyed. In Sample 1 and Sample 2, exploratory and confirmatory factor analysis results confirmed the unidimensional structure of the 10-item abbreviated version of the scale. In Sample 2, Proactive Personality scores were found to be positively correlated with the personality factors of Extraversion, Conscientiousness, Openness, and Agreeableness, and negatively correlated with Neuroticism. In Sample 3, Proactive Personality was found to be positively related to self-efficacy and political skill. In Sample 4, demographics controlled, Proactive Personality explained significant incremental variance in the employees' self-rated career satisfaction and job performance as rated by immediate supervisors. PMID- 19810433 TI - Estimates of general and emotional intelligence for self and parents in Iran. AB - Estimations of IQ and emotional intelligence for self and parents were investigated. Previous studies in both Western and African cultures have found significant sex differences in self-estimates of IQ and emotional intelligence, while IQ was rated higher for fathers than mothers. These prior results suggest the findings should be invariant across culture, and were expected to be replicated here in a predominantly Islamic society with great sociopolitical changes with respect to the Islamic Revolution. 187 Iranian university students estimated their own and their parents' scores on IQ and 15 facets of emotional intelligence on a normal distribution graph. The present results showed no significant sex differences in self-estimates of these variables, while fathers were rated higher on IQ. The implications of these findings are offered in light of sociopolitical changes during the last three decades in Iran. PMID- 19810434 TI - Male sexual coercion: analysis of a few associated factors. AB - The purpose of this study was to assess, in a sample of 700 university students ages 18 to 40 years in El Salvador, the importance of sociodemographic variables (age, having a partner, and frequency of religious practice), personality traits (anger, hostility, and aggressiveness), sexual attitudes, and male chauvinist attitudes (erotophilia, double standard, and rape supportive attitudes) in explaining male sexual coercion. All these variables were assessed. Once the effect of social desirability had been controlled, the analysis showed the importance of trait anger along with a favorable attitude toward the use of violence against women, double standard, and negative attitudes toward sexuality in explaining male sexual coercion. Religious involvement, however, may act as a protective factor against such violence. Multidimensional models are needed to explain sexual violence against women by men in heterosexual relationships. PMID- 19810435 TI - Work-family conflicts and work performance. AB - Prior research indicates that work-family conflict interferes with family far more than it interferes with work. Conservation of resources provides a possible explanation: when shifting resources from family is no longer sufficient to maintain satisfactory work performance, then workers must acquire additional resources or reduce investments in work. One source of such additional resources could be high performance peers in the work group. The performance of workers with resource-rich peers may be less adversely affected by work-family conflict. In this study, 136 employees of a wholesale distribution firm (61% women, 62% minority) working in groups of 7 to 11 in manual labor and low-level administrative jobs rated their own work-to-family conflict. Their supervisors rated workers' performance. Hierarchical regression analysis indicated that work to-family conflict increasingly adversely affected job performance as work group performance decreased. Hence, work group performance may be an important moderator of the effects of work-family conflict. PMID- 19810436 TI - Therapist training, feedback, and practice for dream work: a pilot study. AB - Thirteen therapists (8 women, 5 men; M age = 29.1 yr., SD = 6.3; M counseling experience 2.7 yr., SD = 6.3, range = 1 semester to 8 yr.) received training in the cognitive-experiential model of dream work through a didactic-experiential workshop, individual feedback, and practice. All participants but one were enrolled in masters' or doctoral level counseling training programs. An examination of effect sizes indicated that (a) therapists' self-efficacy about using the dream model increased substantially and positive attitudes toward dreams increased slightly after the didactic-experiential workshop; (b) the process and outcome of the second dream session were better for therapists who received individual feedback after the first session than for therapists who did not receive feedback after the first session; and (c) for the subgroup of six therapists who conducted five sessions, therapists' attitudes toward using the dream model improved, and their self-efficacy for working with dreams and perceived self-competence in dream work increased. PMID- 19810437 TI - Reliability and factor validity of a Farsi version of The Positive and Negative Perfectionism Scale. AB - This study investigated reliability and factor validity of a Farsi version of the Positive and Negative Perfectionism Scale in 606 undergraduate students (257 men, 349 women) from the University of Tehran. All participants were asked to complete the scale, along with the General Health Questionnaire (Goldberg, 1972) and the Coopersmith Self-Esteem Inventory (Coopersmith, 1967). Findings indicated good internal consistency, test-retest reliability, and construct validity of the two factor structure of the Farsi version of the Positive and Negative Perfectionism Scale. These factors were similar to the factors found in previous studies and were accordingly labeled Positive Perfectionism and Negative Perfectionism. The results provide evidence for applicability of the scale and its cross-cultural validity. PMID- 19810438 TI - Childhood emotional neglect related to posttraumatic stress disorder symptoms and body mass index in adult women. AB - The relationships among the severity of childhood abuse and neglect, posttraumatic stress disorder (PTSD), and adult obesity were investigated. 207 women (M age = 26.5 yr., SD = 6.7) completed the Childhood Trauma Questionnaire, Millon Clinician Multiaxial Inventory, and a demographic questionnaire. Analyses of variance indicated that women who reported moderate-to-extreme emotional neglect (n = 71) had significantly higher PTSD scores and increased BMI compared to women who reported low emotional neglect (n = 84). Women who reported severe sexual or emotional abuse also had higher PTSD scores, but no relationship was found with BMI when other factors were controlled. Although PTSD scores and self reported severity of childhood emotional neglect were strongly correlated (r = .61, p < .001), PTSD was not found to be a mediating factor in obesity in women who reported childhood emotional neglect, although depression was. PMID- 19810439 TI - Postpartum mental illness: perspectives from an Arabian Gulf population. AB - There is growing awareness of the importance of mental health issues in the perinatal period in Western societies, but very little information is available from Muslim and Arab countries. Qualitative information gathered using focus group discussions of women of childbearing age is presented along with additional information obtained from key informant interviews with grandmothers, husbands, and health care professionals in the United Arab Emirates. The participants were women attending a public sector clinic in Al Ain, Abu Dhabi, and a private clinic in Dubai on a particular day. There were 19 women in the age range 17 to 46 years (M age = 29 yr.). The majority of the women who took part in the study did not recognize postnatal depression as a psychological issue but considered the problems a result of "evil eye" or "Jinn." The present findings suggest the need for initiating awareness programs among women and training of health professionals on the detection and management of postnatal depression. PMID- 19810440 TI - An attributional sequence model of jury decision making in civil torts. AB - A jury simulation study was designed to replicate and extend prior research on attributional decision-making sequences in a litigation context. Although previous studies implicated injury severity, causality, responsibility, and punishment, a maximum of three stages in various combinations were tested, and only occasionally in the context of civil litigation. The present effort integrated all four stages into a more inclusive model focused on civil jury decision making. Undergraduate participants (N=91) read six case vignettes and responded to attribution questions in each of four categories. Path analyses supported the hypothesized 4-stage attributional sequence. PMID- 19810441 TI - Death anxiety among Ethiopian undergraduate students. AB - Measurement of Death Anxiety among 151 Ethiopian undergraduate students using Templer's scale and Thorson and Powell's scale revealed that the sample has slightly higher than average death anxiety. The results also indicate that in this largely Orthodox Christian sample, students were afraid of the pain in death and less afraid of what happens to their body after death. As some items in each scale did not work well, the Cronbach alphas were low, .61 for the 12 items of Templer's scale and .78 for the 15 items of Thorson and Powell's scale. The correlation between the two full-scale scores was .58 and between scale scores with only acceptable items was .67, indicating the possibility that both scales measure death anxiety equally well if some items are excluded. Results were not consistent with some previous studies in other cultures. Age was significantly related to both Templer's scale (.29) and Thorson and Powell's scale (.28). Death anxiety dimensions like time, control, and afterlife aspects seemed to have doubtful meanings in the Ethiopian sample. PMID- 19810442 TI - Verbal expressions of confidence and doubt. AB - The development of a taxonomy of expressions expressing the degree of confidence or certainty felt in the correctness of one's judgments, knowledge, or beliefs is reported. 30 phrases expressing confidence and doubt were rated by 96 British participants on a 7-point scale to indicate how much confidence or doubt they felt each phrase expressed. The expressions were rank ordered, based on their mean ratings, to produce a continuum of cues expressing confidence, ranging from high to low. 9 of the 30 expressions were rated as expressing lower confidence when phrased in the past tense than in the present tense. The expressions reported in this study form a useful tool for researchers who are investigating the communication of confidence and degrees of belief, especially in relation to giving advice, influence, and persuasion. PMID- 19810443 TI - Attachment type for adolescents in residential treatment centers. AB - The styles of attachment in two groups of adolescents with different characteristics and life experiences are examined. Participants were 81 adolescents residing inside residential treatment centers (M age = 15.5 yr., SD = 1.1; 48% boys, 52% girls) and 81 adolescents living with their own families (M age =15.6 yr., SD = 1.3; 49% boys, 51% girls). The Italian adaptation of the Separation Anxiety Test was used to measure attachment style. Only the data obtained from the "Self" perspective indicated a significant difference between the two groups, showing higher percentages of Secure attachment among adolescents living with their families. When replies were referred to the "Self," the risk of assessing Insecure attachment in the Residential group was about 2 times higher than in the Family group (OR = 2.28). PMID- 19810444 TI - Do warning signs on electronic gaming machines influence irrational cognitions? AB - Electronic gaming machines are popular among problem gamblers; in response, governments have introduced "responsible gaming" legislation incorporating the mandatory display of warning signs on or near electronic gaming machines. These signs are designed to correct irrational and erroneous beliefs through the provision of accurate information on probabilities of winning and the concept of randomness. There is minimal empirical data evaluating the effectiveness of such signs. In this study, 93 undergraduate students were randomly allocated to standard and informative messages displayed on an electronic gaming machine during play in a laboratory setting. Results revealed that a majority of participants incorrectly estimated gambling odds and reported irrational gambling related cognitions prior to play. In addition, there were no significant between group differences, and few participants recalled the content of messages or modified their gambling-related cognitions. Signs placed on electronic gaming machines may not modify irrational beliefs or alter gambling behaviour. PMID- 19810445 TI - Reactions to prayer at governmental meetings. AB - An interaction of mean ratings of support and intent to vote for officials between scenarios in which the religion of a government official offering a prayer was experimentally manipulated, and the reported reaction of the community, was related to 64 college women's support and intention to vote for the official. Importance of religion to the participants contributed significant variance as a covariate. PMID- 19810446 TI - Love styles and desire for closeness in romantic relationships. AB - This study investigated differences in love styles (i.e., eros, ludus, storge, pragma, agape, mania) associated with the romantic desire for closeness. Participants were 197 undergraduate students (M age = 19.8 yr., SD = 1.9; 92 men, 104 women) currently in a romantic relationship who completed a survey assessing their love styles and current desire for closeness with their partner (i.e., desired less closeness, the same level of closeness, or more closeness). Results indicated small significant differences in individuals' preferences for closeness with the eros and ludus love styles. Specifically, individuals who desired less closeness scored lower on eros love and higher on ludus love than partners who reported an ideal level of closeness or who desired more closeness. PMID- 19810447 TI - Psychometric data for a Farsi translation of the Trait Meta-Mood Scale. AB - This study examined the internal consistency, test-retest reliability, and construct validity of a Farsi version of the Trait Meta-Mood Scale, with a sample of 306 undergraduate students (123 men, 183 women) ages 18 to 51 years. Participants completed Farsi versions of the Trait Meta-Mood Scale, the Satisfaction with Life Scale, and the Depression Anxiety Stress Scale. Analysis confirmed the preliminary reliabilities and construct validity of the Trait Meta Mood Scale. PMID- 19810448 TI - The big five traits as predictors of subjective and psychological well-being. AB - Despite considerable research on personality and "hedonic" or subjective well being, parallel research on "eudaimonic" or psychological well-being is scarce. The current study investigated the relationship between the Big Five traits and subjective and psychological well-being among 211 men and women. Results indicated that the relationship between personality factors and psychological well-being was stronger than the relationship between personality factors and subjective well-being. Extraversion, neuroticism, and conscientiousness correlated similarly with both subjective and psychological well-being, suggesting that these traits represent personality predispositions for general well-being. However, the personality correlates of the dimensions within each broad well-being type varied, suggesting that the relationship between personality and well-being is best modeled in terms of associations between specific traits and well-being dimensions. PMID- 19810449 TI - Impairment in the recognition of facial expression: a case study. AB - In this study, a case (HY) is described. This man, now 25 yr. old, lived in a persistent vegetative state for 6 yr. after encephalitis at the age of 10 yr. He was reportedly impaired at recognizing fear, and in everyday life, apparently had impaired recognition of anger as well. In testing with facial expressions, no obvious differences between HY and normal controls in anger perceptions were found. In this study, Japanese and Caucasian models of facial expression were used; on these tests, HY was impaired at recognizing facial expressions of anger only in the Japanese models. PMID- 19810450 TI - Validation of the Personal Need for Structure Scale in Chinese. AB - To validate the Chinese version of the Personal Need for Structure Scale, questionnaires were administered to 1,418 individuals in three samples. Item total correlations and internal consistency of the scale were acceptable. The test-retest reliability was .79. Confirmatory factor analysis indicated that the Chinese version comprised two dimensions, as did the original version; Desire for Structure and Response to Lack of Structure. Correlation coefficients between the Personal Need for Structure Scale and other related measures indicated that the scale has acceptable discriminant validity and convergent validity. PMID- 19810451 TI - Psychometric properties of Chen, Gully, and Eden's "New General Self-efficacy Scale" in a Greek sample. AB - The aim of the present study was to translate and investigate the psychometric properties of the New General Self-Efficacy Scale with a group of 551 Greek children (269 girls, 282 boys; ages 9 to 12 years, Grades 4, 5, and 6). A confirmatory factor analysis was performed which verified the one-factor model. The reliability, validity, and item analysis appeared to be adequate. PMID- 19810452 TI - Youth religiosity and substance use: a meta-analysis from 1995 to 2007. AB - In this meta-analysis, the magnitude of the protective effects of religiosity on youth involvement in substance use was investigated. Based on 22 studies in peer reviewed journals published between 1995 and 2007, the average weighted mean correlation was Zr = .16, significant regardless of the definitions of religiosity. The homogeneity test of variance showed consistent protective effects of religiosity on four types of substance use, namely, alcohol, cigarette, marijuana, and other illicit drugs. PMID- 19810453 TI - Are women in China sexist toward other women? A study of Chinese college students. AB - This study assessed the extent to which social sexism affects Chinese women's perception and evaluation of other women's performance. A sample of 100 college women was selected in a top university in Beijing, China, and was asked to read six scholastic essays and then evaluate the quality of the essays and competence of the authors. Male and female names were randomly assigned as authors of the essays, and the respondents were blind to the arrangement. Results showed that the essays assumed to be written by male authors did not receive higher scores than those assumed to be written by female authors on quality or competence items. Sexism is not marked among these highly educated young women. PMID- 19810454 TI - Future time orientation and temperament: exploration of their relationship to primary and secondary psychopathy. AB - The present study combines Lykken's theory about the role of reward sensitivity and punishment insensitivity in the development of antisocial behavior with Gjesme's theory of future time orientation. 158 adolescents comprised a target group of 79 adolescents who had defined behavioral problems and a matched referential group of 79 adolescents who did not have notable behavioral problems. The results suggest that attributes related to primary psychopathy are associated with a relatively weak or hyporeactive behavioral inhibition system, behavioral approach reactivity, and low future time orientation. Moreover, attributes related to secondary psychopathy are related to an overly sensitive (hyper reactive) behavioral approach system and low future time orientation. Robust positive associations for behavioral approach reactivity and low future time orientation with primary and secondary psychopathy suggest that high behavioral approach/low future time orientation may represent a core feature common to the two factors of psychopathy. PMID- 19810455 TI - Roles of participation and feedback in group potency. AB - The roles of group participation and group performance feedback were examined as antecedents of group potency, i.e., beliefs shared among a work group's members about the general effectiveness of the work group. Also examined were how group participation and the congruence of the feedback received from different sources about performance predicted convergence in members' beliefs about group effectiveness. The sample comprised 61 work groups of professionals involved in Master in Business Administration (MBA) programs (284 participants). Mean group size was 4.6 members (SD = .58). 65% of participants were male, and 51% were between 30 and 40 years of age. Data were gathered at two measurement times. Increases in group participation were positively related to increases in group potency and the convergence in beliefs about group effectiveness among group members over time. Results supported the premise that group performance feedback is an antecedent of changes in group potency over time. PMID- 19810456 TI - General George S. Patton, Jr. and the conquest of fear. AB - General George S. Patton, Jr. was a highly successful World War II battle commander whose flamboyance and many idiosyncrasies made him a focus of interest for biographers. But he was an enigmatic and complex man whose success came at a high price. Despite his prominence and celebrity, there have been minimal efforts to examine his psychological makeup so crucial to his success on the battlefield. In this essay, Patton's personal story and how it relates to the stresses of war and to his leadership of men in arms is examined. Central to his success was his early triumph over dyslexia, his ability to control the fear and guilt inherent in combat, his intense physical activity, his theatrical skills, and his deep knowledge of the history and methods of warfare. PMID- 19810457 TI - Point/counterpoint. The chief information technology officer in a radiation oncology department should be a medical physicist. PMID- 19810458 TI - An easy method to account for light scattering dose dependence in radiochromic films. AB - PURPOSE: To date no detector can offer the unbeatable characteristics of film dosimetry in terms of spatial resolution and this is why it has been chosen by many institutions for treatment verification and, in that respect, radiochromic films are becoming increasingly popular due to their advantageous properties. It is the aim of this work to suggest an easy method to overcome one of the drawbacks in radiochromic film dosimetry associated with the scanning device, namely, the nonuniform dose dependent response, mainly due to the light scattering effect. METHODS: The suggested procedure consists of building four correction matrices by sequentially scanning one, two, three, and four unexposed blank films. The color level of these four matrices is compatible with four points in the calibration curve dose range. Therefore, the dose dependent correction to the scanned irradiated film will be obtained by interpolating between the four correction matrices. RESULTS: The validity of the suggested method is checked against an ion chamber 2D array. The use of the proposed flattening correction improves considerably the dose agreement when compared with the cases in which no correction is applied. CONCLUSIONS: The method showed to be fast and easy and practically overcomes the dependence on the dose of light scattering of flatbed scanners. PMID- 19810459 TI - EGSnrc-based Monte Carlo dosimetry of CSA1 and CSA2 137Cs brachytherapy source models. AB - PURPOSE: AAPM TG-56 recommends the use of a specific dosimetric dataset for each brachytherapy source model. In this study, a full dosimetric dataset for indigenously developed 137Cs source models, namely, the CSA1 and CSA2, in accordance with the AAPM TG-43U1 formalism is presented. The study includes calculation of dose-to-kerma ratio D/K in water around these sources including stainless steel encapsulated 137Cs sources such as RTR, 3M, and selectron/LDR 137Cs. METHODS: The Monte Carlo-based EGSnrcMP code system is employed for modeling the sources in vacuum and in water. Calculations of air-kerma strength, S(K) for the investigated sources and collision kerma in water along the transverse axis of the RTR source are based on the FLURZnrc code. Simulations of water-kerma and dose in water for the CSA1, CSA2, RTR, 3M, and selectron/ LDR 137Cs sources are carried out using the DOSRZnrc code. In DOSRZnrc calculations, water-kerma and dose are scored in a cylindrical water phantom having dimensions of 80 cm diameter x 80 cm height. RESULTS: The calculated dose-rate constants for the CSA1 and CSA2 sources are 0.945(1) and 1.023(1) cGy/(h U), respectively. The calculated value of S(K) per unit source activity, S(K)/A for the CSA1 and CSA2 sources is 7.393(7) x 10(-8) cGy cm2/(h Bq). The EGSnrcMP-based collision kerma rates for the RTR source along the transverse axis (0.25-10 cm) agree with the corresponding GEANT4-based published values within 0.5%. Anisotropy profiles of the CSA1 and CSA2 sources are significantly different from those of other sources. For the selectron/LDR single pellet 137Cs spherical source (modeled as a cylindrical pellet with dimensions similar to the seed selectron), the values of D/K at 1 and 1.25 mm from the capsule are 1.023(1) and 1.029(1), respectively. The value of D/K at 1 mm from the CSA1, CSA2, RTR, and 3M 137Cs source capsules (all sources have an external radius of 1.5 mm) is 1.017(1) and this ratio is applicable to axial positions z = 0 to z = -L/2. This is in contrast to a published GEANT4-based Monte Carlo dosimetric study on RTR and 3M 137Cs sources wherein the authors have assumed that collision kerma is approximately equal to absorbed dose at 1 mm from the source capsules. Collision kerma is approximately equal to absorbed dose for distances > or = 2 mm from source capsules as opposed to > or = 1 mm reported in published studies. A detailed electron transport is necessary up to 2 mm from source capsules. CONCLUSIONS: The Monte Carlo calculated dose-rate data for the CSA1 and CSA2 sources can be used as input data for treatment planning or to verify the calculations by radiotherapy treatment planning system. PMID- 19810461 TI - An absolute dose determination of helical tomotherapy accelerator, TomoTherapy High-Art II. AB - PURPOSE: A helical tomotherapy accelerator presents a dosimetric challenge because, to this day, there is no internationally accepted protocol for the determination of the absolute dose. Because of this reality, we investigated the different alternatives for characterizing and measuring the absolute dose of such an accelerator. We tested several dosimetric techniques with various metrological traceabilities as well as using a number of phantoms in static and helical modes. METHODS: Firstly, the relationship between the reading of ionization chambers and the absorbed dose is dependent on the beam quality value of the photon beam. For high energy photons, the beam quality is specified by the tissue phantom ratio (TPR20,10) and it is therefore necessary to know the TPR20,10 to calculate the dose delivered by a given accelerator. This parameter is obtained through the ratio of the absorbed dose at 20 and 10 cm depths in water and was measured in the particular conditions of the tomotherapy accelerator. Afterward, measurements were performed using the ionization chamber (model A1SL) delivered as a reference instrument by the vendor. This chamber is traceable in absorbed dose to water in a Co-60 beam to a water calorimeter of the American metrology institute (NIST). Similarly, in Switzerland, each radiotherapy department is directly traceable to the Swiss metrology institute (METAS) in absorbed dose to water based on a water calorimeter. For our research, this traceability was obtained by using an ionization chamber traceable to METAS (model NE 2611A), which is the secondary standard of our institute. Furthermore, in order to have another fully independent measurement method, we determined the dose using alanine dosimeters provided by and traceable to the British metrology institute (NPL); they are calibrated in absorbed dose to water using a graphite calorimeter. And finally, we wanted to take into account the type of chamber routinely used in clinical practice and therefore measured the dose using a Farmer-type instrument (model NE 2571) as well. RESULTS: We found the tomotherapy TPR20,10 value to be around 0.629, which is close to a 4 MV conventional linear accelerator value. During static irradiation, the secondary standard and the alanine dosimeters were compatible within 0.5%. The A1SL relative deviation to the secondary standard was 1.2% and the NE2571 relative deviation to the secondary standard was -1.7%. The measurement in dynamic helical mode found the different dosimeters compatible within 1.4% and the alanine dosimeters and the secondary standard were even found under 0.2%. CONCLUSIONS: We found that the different methods are all within uncertainties as well as globally coherent, and the specific limitations of the various dosimeters are discussed in order to help the medical physicist design an independent reference system. We demonstrated that, taking into account the particular reference conditions, one can use an ionization chamber calibrated for conventional linear accelerators to assert the absolute dose delivered by a tomotherapy accelerator. PMID- 19810460 TI - Comparisons of treatment optimization directly incorporating random patient setup uncertainty with a margin-based approach. AB - The purpose of this study is to incorporate the dosimetric effect of random patient positioning uncertainties directly into a commercial treatment planning system's IMRT plan optimization algorithm through probabilistic treatment planning (PTP) and compare coverage of this method with margin-based planning. In this work, PTP eliminates explicit margins and optimizes directly on the estimated integral treatment dose to determine optimal patient dose in the presence of setup uncertainties. Twenty-eight prostate patient plans adhering to the RTOG-0126 criteria are optimized using both margin-based and PTP methods. Only random errors are considered. For margin-based plans, the planning target volume is created by expanding the clinical target volume (CTV) by 2.1 mm to accommodate the simulated 3 mm random setup uncertainty. Random setup uncertainties are incorporated into IMRT dose evaluation by convolving each beam's incident fluence with a sigma = 3 mm Gaussian prior to dose calculation. PTP optimization uses the convolved fluence to estimate dose to ensure CTV coverage during plan optimization. PTP-based plans are compared to margin-based plans with equal CTV coverage in the presence of setup errors based on dose volume metrics. The sensitivity of the optimized plans to patient-specific setup uncertainty variations is assessed by evaluating dose metrics for dose distributions corresponding to halving and doubling of the random setup uncertainty used in the optimization. Margin-based and PTP-based plans show similar target coverage. A physician review shows that PTP is preferred for 21 patients, margin-based plans are preferred in 2 patients, no preference is expressed for 1 patient, and both autogenerated plans are rejected for 4 patients. For the PTP-based plans, the average CTV receiving the prescription dose decreases by 0.5%, while the mean dose to the CTV increases by 0.7%. The CTV tumor control probability (TCP) is the same for both methods with the exception of one case in which PTP gave a slightly higher TCP. For critical structures that do not meet the optimization criteria, PTP shows a decrease in the volume receiving the maximum specified dose. PTP reduces local normal tissue volumes receiving the maximum dose on average by 48%. PTP results in lower mean dose to all critical structures for all plans. PTP results in a 2.5% increase in the probability of uncomplicated control (P+), along with a 1.9% reduction in rectum normal tissue complication probability (NTCP), and a 0.7% reduction in bladder NTCP. PTP-based plans show improved conformality as compared with margin-based plans with an average PTP-based dosimetric margin at 7100 cGy of 0.65 cm compared with the margin-based 0.90 cm and a PTP-based dosimetric margin at 3960 cGy of 1.60 cm compared with the margin-based 1.90 cm. PTP-based plans show similar sensitivity to variations of the uncertainty during treatment from the uncertainty used in planning as compared to margin-based plans. For equal target coverage, when compared to margin-based plans, PTP results in equal or lower doses to normal structures. PTP results in more conformal plans than margin-based plans and shows similar sensitivity to variations in uncertainty. PMID- 19810462 TI - Monte Carlo analysis of energy dependent anisotropy of bremsstrahlung x-ray spectra. AB - The energy resolved emission angle dependence of x-ray spectra was analyzed by MCNPX (Monte Carlo N particle Monte Carlo) simulator. It was shown that the spectral photon flux had a maximum at a well-defined emission angle due to the anisotropy of the bremsstrahlung process. The higher the relative photon energy, the smaller the emission angle belonging to the maximum was. The trends predicted by the Monte Carlo simulations were experimentally verified. The Monte Carlo results were compared to both the Institute of Physics and Engineering in Medicine spectra table and the SPEKCALCV1.0 code. PMID- 19810463 TI - Considering marker visibility during leaf sequencing for segmental intensity modulated radiation therapy. AB - PURPOSE: Segmental intensity-modulated radiation therapy (IMRT) delivers a sequence of segments to obtain a desired intensity distribution. Many leaf sequencing algorithms for segmental IMRT have been developed with the aim of reducing delivered monitor units (MUs) and (or) number of segments and, consequently, to reduce the total treatment delivery time. With the development of real-time detection technology, it is useful to develop leaf sequencing algorithms that consider the detecting probability of markers implanted into or near the target volume. METHODS: In this study, the authors defined the concept of marker visibility to denote the marker's detecting probability and proposed a new leaf sequencing algorithm based on the Kamath algorithm. The new algorithm first uses the Kamath algorithm to generate an initial leaf sequence and then performs a series of column transformations to obtain a new leaf sequence that is optimal in terms of MU efficiency and marker visibility. The authors evaluated the performance of the new algorithm with six artificial fields that had randomly generated intensity matrices and 15 clinical fields that had intensity matrices from the IMRT plans for three prostate cancer patients. RESULTS: Compared to the Kamath algorithm, the new algorithm does not increase the total delivered intensity but increases the marker visibility. For the artificial fields, the marker visibility increased from 66.67% to 91.67% for small (5 x 5) radiation fields, from 39.29% to 42.86% for medium size (10 x 10) fields, and from 31.48% to 37.04% for large (20 x 20) fields. For the clinical fields, the marker visibility increased 9%-20% for four fields, 20%-30% for three fields, 30%-40% for two fields, and more than 40% for one field. However, the marker visibility did not change for 4 out of 15 fields. CONCLUSIONS: The authors developed a new leaf sequencing algorithm for optimal MU efficiency and marker visibility and also rigorously proved its optimality. PMID- 19810464 TI - A two-step scheme for distortion rectification of magnetic resonance images. AB - The aim of this work is to demonstrate a complete, robust, and time-efficient method for distortion correction of magnetic resonance (MR) images. It is well known that MR images suffer from both machine-related spatial distortions [gradient nonlinearity and main field (B0) inhomogeneity] and patient-related spatial distortions (susceptibility and chemical shift artifacts), and growing interest in the area of MR-based radiotherapy treatment planning has put new requirements on the geometric accuracy of such images. The authors present a two step method that combines a phantom-based reverse gradient technique for measurement of gradient nonlinearities and a patient-based phase difference mapping technique for measurement of B0 inhomogeneities, susceptibility, and chemical shift distortions. The phase difference mapping technique adds only minutes to the total patient scan time and can be used to correct a variety of images of the same patient and anatomy. The technique was tested on several different phantoms, each designed to isolate one type of distortion. The mean distortion was reduced to 0.2 +/- 0.1 mm in both gradient echo and spin echo images of a grid phantom. For the more difficult case of a highly distorted echo planar image, residual distortion was reduced to subvoxel dimensions. As a final step, the technique was implemented on patient images. The current technique is effective, time efficient, and robust and provides promise for preparing distortion-rectified MR images for use in MR-based treatment planning. PMID- 19810465 TI - A hierarchical spectral clustering and nonlinear dimensionality reduction scheme for detection of prostate cancer from magnetic resonance spectroscopy (MRS). AB - Magnetic resonance spectroscopy (MRS) has been shown to have great clinical potential as a supplement to magnetic resonance imaging in the detection of prostate cancer (CaP). MRS provides functional information in the form of changes in the relative concentration of specific metabolites including choline, creatine, and citrate which can be used to identify potential areas of CaP. With a view to assisting radiologists in interpretation and analysis of MRS data, some researchers have begun to develop computer-aided detection (CAD) schemes for CaP identification from spectroscopy. Most of these schemes have been centered on identifying and integrating the area under metabolite peaks which is then used to compute relative metabolite ratios. However, manual identification of metabolite peaks on the MR spectra, and especially via CAD, is a challenging problem due to low signal-to-noise ratio, baseline irregularity, peak overlap, and peak distortion. In this article the authors present a novel CAD scheme that integrates nonlinear dimensionality reduction (NLDR) with an unsupervised hierarchical clustering algorithm to automatically identify suspicious regions on the prostate using MRS and hence avoids the need to explicitly identify metabolite peaks. The methodology comprises two stages. In stage 1, a hierarchical spectral clustering algorithm is used to distinguish between extracapsular and prostatic spectra in order to localize the region of interest (ROI) corresponding to the prostate. Once the prostate ROI is localized, in stage 2, a NLDR scheme, in conjunction with a replicated clustering algorithm, is used to automatically discriminate between three classes of spectra (normal appearing, suspicious appearing, and indeterminate). The methodology was quantitatively and qualitatively evaluated on a total of 18 1.5 T in vivo prostate T2-weighted (w) and MRS studies obtained from the multisite, multi-institutional American College of Radiology (ACRIN) trial. In the absence of the precise ground truth for CaP extent on the MR imaging for most of the ACRIN studies, probabilistic quantitative metrics were defined based on partial knowledge on the quadrant location and size of the tumor. The scheme, when evaluated against this partial ground truth, was found to have a CaP detection sensitivity of 89.33% and specificity of 79.79%. The results obtained from randomized threefold and fivefold cross validation suggest that the NLDR based clustering scheme has a higher CaP detection accuracy compared to such commonly used MRS analysis schemes as z score and PCA. In addition, the scheme was found to be robust to changes in system parameters. For 6 of the 18 studies an expert radiologist laboriously labeled each of the individual spectra according to a five point scale, with 1/2 representing spectra that the expert considered normal and 3/4/5 being spectra the expert deemed suspicious. When evaluated on these expert annotated datasets, the CAD system yielded an average sensitivity (cluster corresponding to suspicious spectra being identified as the CaP class) and specificity of 81.39% and 64.71%, respectively. PMID- 19810466 TI - Technical note: RabbitCT--an open platform for benchmarking 3D cone-beam reconstruction algorithms. AB - PURPOSE: Fast 3D cone beam reconstruction is mandatory for many clinical workflows. For that reason, researchers and industry work hard on hardware optimized 3D reconstruction. Backprojection is a major component of many reconstruction algorithms that require a projection of each voxel onto the projection data, including data interpolation, before updating the voxel value. This step is the bottleneck of most reconstruction algorithms and the focus of optimization in recent publications. A crucial limitation, however, of these publications is that the presented results are not comparable to each other. This is mainly due to variations in data acquisitions, preprocessing, and chosen geometries and the lack of a common publicly available test dataset. The authors provide such a standardized dataset that allows for substantial comparison of hardware accelerated backprojection methods. METHODS: They developed an open platform RabbitCT (www.rabbitCT.com) for worldwide comparison in backprojection performance and ranking on different architectures using a specific high resolution C-arm CT dataset of a rabbit. This includes a sophisticated benchmark interface, a prototype implementation in C++, and image quality measures. RESULTS: At the time of writing, six backprojection implementations are already listed on the website. Optimizations include multithreading using Intel threading building blocks and OpenMP, vectorization using SSE, and computation on the GPU using CUDA 2.0. CONCLUSIONS: There is a need for objectively comparing backprojection implementations for reconstruction algorithms. RabbitCT aims to provide a solution to this problem by offering an open platform with fair chances for all participants. The authors are looking forward to a growing community and await feedback regarding future evaluations of novel software- and hardware-based acceleration schemes. PMID- 19810467 TI - Measurements of dose discrepancies due to inhomogeneities and radiographic contrast in balloon catheter brachytherapy. AB - Recently, a device called MammoSite, consisting of a balloon and a catheter, was developed to perform partial-breast irradiation using a high-dose-rate (HDR) brachytherapy unit with ease and reproducibility. However, the actual dose to the skin does not agree well with the calculated dose by the treatment planning system because of the difference between the calculation condition and the real treatment condition (i.e., homogeneous water and full scatter condition vs contrast solution and lack of full scatter condition). In this study, the authors experimentally estimated dose discrepancies due to contrast and lack of full scatter in breast HDR brachytherapy with MammoSite. Using metal-oxide semiconductor field-effect transistor detectors and a breast phantom, the dose discrepancies between the calculation and the treatment conditions were measured according to contrast concentration (10% and 20% volume ratios), balloon size (35 and 60 cm3), and source to detector distance ranging from 25 to 50 mm. The source was an Ir-192 isotope from Nucletron HDR unit. The dose discrepancies from the calculation condition due to both contrast and lack of full scatter combined ranged from about -1.4 +/- 2.5% to -18.2 +/- 2.0% in the studied cases (error bound is in two sided confidence interval of 80% based on Student's t distribution). In all cases, the effect of lack of full scatter was dominant to that of contrast and significant dose discrepancies existed between the calculation and the real treatment conditions, indicating that the actual skin dose is less than that which is currently calculated. PMID- 19810468 TI - Megavoltage planar and cone-beam imaging with low-Z targets: dependence of image quality improvement on beam energy and patient separation. AB - PURPOSE: The purpose of this study is to investigate the improvement of megavoltage planar and cone-beam CT (CBCT) image quality with the use of low atomic number (Z) external targets in the linear accelerator. METHODS: In this investigation, two experimental megavoltage imaging beams were generated by using either 3.5 or 7.0 MeV electrons incident on aluminum targets installed above the level of the carousel in a linear accelerator (2100EX, Varian Medical, Inc., Palo Alto, CA). Images were acquired using an amorphous silicon detector panel. Contrast-to-noise ratio (CNR) in planar and CBCT images was measured as a function of dose and a comparison was made between the imaging beams and the standard 6 MV therapy beam. Phantoms of variable diameter were used to examine the loss of contrast due to beam hardening. Porcine imaging was conducted to examine qualitatively the advantages of the low-Z target approach in CBCT. RESULTS: In CBCT imaging CNR increases by factors as high as 2.4 and 4.3 for the 7.0 and 3.5 MeV/Al beams, respectively, compared to images acquired with 6 MV. Similar factors of improvement are observed in planar imaging. For the imaging beams, beam hardening causes a significant loss of the contrast advantage with increasing phantom diameter; however, for the 3.5 MeV/Al beam and a phantom diameter of 25 cm, a contrast advantage remains, with increases of contrast by factors of 1.5 and 3.4 over 6 MV for bone and lung inhale regions, respectively. The spatial resolution is improved slightly in CBCT images for the imaging beams. CBCT images of a porcine cranium demonstrate qualitatively the advantages of the low-Z target approach, showing greater contrast between tissues and improved visibility of fine detail. CONCLUSIONS: The use of low-Z external targets in the linear accelerator improves megavoltage planar and CBCT image quality significantly. CNR may be increased by a factor of 4 or greater. Improvement of the spatial resolution is also apparent. PMID- 19810469 TI - Multiple scattering of 13 and 20 MeV electrons by thin foils: a Monte Carlo study with GEANT, Geant4, and PENELOPE. AB - PURPOSE: In this work, recent results from experiments and simulations (with EGSnrc) performed by Ross et al. [Med. Phys. 35, 4121-4131 (2008)] on electron scattering by foils of different materials and thicknesses are compared to those obtained using several Monte Carlo codes. METHODS: Three codes have been used: GEANT (version 3.21), Geant4 (version 9.1, patch03), and PENELOPE (version 2006). In the case of PENELOPE, mixed and fully detailed simulations have been carried out. RESULTS: Transverse dose distributions in air have been obtained in order to compare with measurements. The detailed PENELOPE simulations show excellent agreement with experiment. The calculations performed with GEANT and PENELOPE (mixed) agree with experiment within 3% except for the Be foil. In the case of Geant4, the distributions are 5% narrower compared to the experimental ones, though the agreement is very good for the Be foil. Transverse dose distribution in water obtained with PENELOPE (mixed) is 4% wider than those calculated by Ross et al. using EGSnrc and is 1% narrower than the transverse dose distributions in air, as considered in the experiment. CONCLUSIONS: All the codes give a reasonable agreement (within 5%) with the experimental results for all the material and thicknesses studied. PMID- 19810470 TI - An evaluation of ionization chambers for the relative dosimetry of kilovoltage x ray beams. AB - In this work, the authors have evaluated ten different ionization chambers for the relative dosimetry of kilovoltage x-ray beams in the energy range of 50-280 kVp. Percentage depth doses in water and relative detector response (in Solid Water and in air) were measured for each of the x-ray beams studied using a number of chambers. Measured depth dose data were compared with Monte Carlo calculated depth doses using the EGSnrc Monte Carlo package and the BEAMnrc user code. The accuracy of the phase space files generated by BEAMnrc was verified by calculating the half-value layer and comparing with the measured half-value layer of each x-ray beam. The results indicate that the Advanced Markus, Markus, NACP, and Roos parallel plate ionization chambers were suitable for the measurement of depth dose data in this beam quality range with an uncertainty of less than 3%, including in the regions close to the water surface. While the relative detector response of the Farmer and scanning thimble chambers exhibited a better energy response, they were not suitable for depth dose measurements in the first 5 mm below the water surface with differences of up to 12% in the surface dose measurement for the 50 kVp x-ray beam. These differences were due to dose artifacts generated by the chamber size and the dose gradient. However, at depths greater than 5 mm, the Farmer and thimble scanning chambers gave uncertainties of less than 3% for the depth dose measurements for all beam energies. The PTW PinPoint 31006 chamber was found to give varying dose differences of up to 8% depending on the x-ray beam energy; this was attributed to the steel central electrode. The authors recommend that one of the parallel plate ionization chambers investigated be used to determine depth dose data for kilovoltage x-ray beams in the energy range studied and give correct dose information close to the surface and at depth in the water phantom. PMID- 19810471 TI - A CT-based analytical dose calculation method for HDR 192Ir brachytherapy. AB - PURPOSE: This article presents an analytical dose calculation method for high dose-rate 192Ir brachytherapy, taking into account the effects of inhomogeneities and reduced photon backscatter near the skin. The adequacy of the Task Group 43 (TG-43) two-dimensional formalism for treatment planning is also assessed. METHODS: The proposed method uses material composition and density data derived from computed tomography images. The primary and scatter dose distributions for each dwell position are calculated first as if the patient is an infinite water phantom. This is done using either TG-43 or a database of Monte Carlo (MC) dose distributions. The latter can be used to account for the effects of shielding in water. Subsequently, corrections for photon attenuation, scatter, and spectral variations along medium- or low-Z inhomogeneities are made according to the radiological paths determined by ray tracing. The scatter dose is then scaled by a correction factor that depends on the distances between the point of interest, the body contour, and the source position. Dose calculations are done for phantoms with tissue and lead inserts, as well as patient plans for head-and neck, esophagus, and MammoSite balloon breast brachytherapy treatments. Gamma indices are evaluated using a dose-difference criterion of 3% and a distance-to agreement criterion of 2 mm. PTRAN_CT MC calculations are used as the reference dose distributions. RESULTS: For the phantom with tissue and lead inserts, the percentages of the voxels of interest passing the gamma criteria (Pgamma > or = 1) are 100% for the analytical calculation and 91% for TG-43. For the breast patient plan, TG-43 overestimates the target volume receiving the prescribed dose by 4% and the dose to the hottest 0.1 cm3 of the skin by 9%, whereas the analytical and MC results agree within 0.4%. Pgamma > or = 1 are 100% and 48% for the analytical and TG-43 calculations, respectively. For the head-and-neck and esophagus patient plans, Pgamma > or = 1 are > or = 99% for both calculation methods. CONCLUSIONS: A correction-based dose calculation method has been validated for HDR 192Ir brachytherapy. Its high calculation efficiency makes it feasible for use in treatment planning. Because tissue inhomogeneity effects are small and primary dose predominates in the near-source region, TG-43 is adequate for target dose estimation provided shielding and contrast solution are not used. PMID- 19810472 TI - A comparison of HDR brachytherapy and IMRT techniques for dose escalation in prostate cancer: a radiobiological modeling study. AB - A course of one to three large fractions of high dose rate (HDR) interstitial brachytherapy is an attractive alternative to intensity modulated radiation therapy (IMRT) for delivering boost doses to the prostate in combination with additional external beam irradiation for intermediate risk disease. The purpose of this work is to quantitatively compare single-fraction HDR boosts to biologically equivalent fractionated IMRT boosts, assuming idealized image guided delivery (igIMRT) and conventional delivery (cIMRT). For nine prostate patients, both seven-field IMRT and HDR boosts were planned. The linear-quadratic model was used to compute biologically equivalent dose prescriptions. The cIMRT plan was evaluated as a static plan and with simulated random and setup errors. The authors conclude that HDR delivery produces a therapeutic ratio which is significantly better than the conventional IMRT and comparable to or better than the igIMRT delivery. For the HDR, the rectal gBEUD analysis is strongly influenced by high dose DVH tails. A saturation BED, beyond which no further injury can occur, must be assumed. Modeling of organ motion uncertainties yields mean outcomes similar to static plan outcomes. PMID- 19810473 TI - Dual resolution cone beam breast CT: a feasibility study. AB - PURPOSE: In this study, the authors investigated the feasibility of a dual resolution volume-of-interest (VOI) cone beam breast CT technique and compared two implementation approaches in terms of dose saving and scatter reduction. METHODS: With this technique, a lead VOI mask with an opening is inserted between the x-ray source and the breast to deliver x-ray exposure to the VOI while blocking x rays outside the VOI. A CCD detector is used to collect the high resolution projection data of the VOI. Low resolution cone beam CT (CBCT) images of the entire breast, acquired with a flat panel (FP) detector, were used to calculate the projection data outside the VOI with the ray-tracing reprojection method. The Feldkamp-Davis-Kress filtered backprojection algorithm was used to reconstruct the dual resolution 3D images. Breast phantoms with 180 microm and smaller microcalcifications (MCs) were imaged with both FP and FP-CCD dual resolution CBCT systems, respectively. Two approaches of implementing the dual resolution technique, breast-centered approach and VOI-centered approach, were investigated and evaluated for dose saving and scatter reduction with Monte Carlo simulation using a GEANT4 package. RESULTS: The results showed that the breast centered approach saved more breast absorbed dose than did VOI-centered approach with similar scatter reduction. The MCs in fatty breast phantom, which were invisible with FP CBCT scan, became visible with the FP-CCD dual resolution CBCT scan. CONCLUSIONS: These results indicate potential improvement of the image quality inside the VOI with reduced breast dose both inside and outside the VOI. PMID- 19810474 TI - From the limits of the classical model of sensitometric curves to a realistic model based on the percolation theory for GafChromic EBT films. AB - PURPOSE: Modern radiotherapy uses complex treatments that necessitate more complex quality assurance procedures. As a continuous medium, GafChromic EBT films offer suitable features for such verification. However, its sensitometric curve is not fully understood in terms of classical theoretical models. In fact, measured optical densities and those predicted by the classical models differ significantly. This difference increases systematically with wider dose ranges. Thus, achieving the accuracy required for intensity-modulated radiotherapy (IMRT) by classical methods is not possible, plecluding their use. As a result, experimental parametrizations, such as polynomial fits, are replacing phenomenological expressions in modern investigations. This article focuses on identifying new theoretical ways to describe sensitometric curves and on evaluating the quality of fit for experimental data based on four proposed models. METHODS: A whole mathematical formalism starting with a geometrical version of the classical theory is used to develop new expressions for the sensitometric curves. General results from the percolation theory are also used. A flat-bed-scanner-based method was chosen for the film analysis. Different tests were performed, such as consistency of the numeric results for the proposed model and double examination using data from independent researchers. RESULTS: Results show that the percolation-theory-based model provides the best theoretical explanation for the sensitometric behavior of GafChromic films. The different sizes of active centers or monomer crystals of the film are the basis of this model, allowing acquisition of information about the internal structure of the films. Values for the mean size of the active centers were obtained in accordance with technical specifications. In this model, the dynamics of the interaction between the active centers of GafChromic film and radiation is also characterized by means of its interaction cross-section value. CONCLUSIONS: The percolation model fulfills the accuracy requirements for quality-control procedures when large ranges of doses are used and offers a physical explanation for the film response. PMID- 19810475 TI - Secondary neutron spectra from modern Varian, Siemens, and Elekta linacs with multileaf collimators. AB - Neutrons are a by-product of high-energy x-ray radiation therapy (threshold for [gamma,n] reactions in high-Z material -7 MeV). Neutron production varies depending on photon beam energy as well as on the manufacturer of the accelerator. Neutron production from modern linear accelerators (linacs) has not been extensively compared, particularly in terms of the differences in the strategies that various manufacturers have used to implement multileaf collimators (MLCs) into their linac designs. However, such information is necessary to determine neutron dose equivalents for different linacs and to calculate vault shielding requirements. The purpose of the current study, therefore, was to measure the neutron spectra from the most up-to-date linacs from three manufacturers: Varian 21EX operating at 15, 18, and 20 MV, Siemens ONCOR operating at 15 and 18 MV, and Elekta Precise operating at 15 and 18 MV. Neutron production was measured by means of gold foil activation in Bonner spheres. Based on the measurements, the authors determined neutron spectra and calculated the average energy, total neutron fluence, ambient dose equivalent, and neutron source strength. The shapes of the neutron spectra did not change significantly between accelerators or even as a function of treatment energy. However, the neutron fluence, and therefore the ambient dose equivalent, did vary, increasing with increasing treatment energy. For a given nominal treatment energy, these values were always highest for the Varian linac. The current study thus offers medical physicists extensive information about the neutron production of MLC-equipped linacs currently in operation and provides them information vital for accurate comparison and prediction of neutron dose equivalents and calculation of vault shielding requirements. PMID- 19810476 TI - Effects of tertiary MLC configuration on secondary neutron spectra from 18 MV x ray beams for the Varian 21EX linear accelerator. AB - The effect of the jaw configuration and the presence and configuration of the tertiary multileaf collimator (MLC) on the secondary neutron spectra for an 18 MV Varian 21EX linear accelerator (linac) is investigated in detail. The authors report the measured spectra for four collimator (jaw-and-MLC) configurations. These configurations represent the extreme settings of the jaws and MLC and should therefore describe the range of possible fluence and spectra that may be encountered during use of this linac. In addition to measurements, a Monte Carlo model was used to simulate the four collimator configurations and calculate the energy spectra and fluence at the same location as it was measured. The Monte Carlo model was also used to calculate the sources of neutron production in the linac head for each collimator configuration. They found that photoneutron production in the linac treatment head is dominated by the order in which the primary photon beam intercepts the high-Z material. The primary collimator, which has the highest position in the linac head (in a fixed location), is the largest source of secondary neutrons. Thereafter, the collimator configuration plays a role in where the neutrons originate. For instance, if the jaws are closed, they intercept the beam and contribute substantially to the secondary neutron production. Conversely, if the jaws are open, the MLC plays a larger role in neutron production (assuming, of course, that it intercepts the beam). They found that different collimator configurations make up to a factor of 2 difference in the ambient dose equivalent. PMID- 19810477 TI - Scintillator avalanche photoconductor with high resolution emitter readout for low dose x-ray imaging: lag. AB - PURPOSE: A new concept of indirect conversion flat-panel imager with avalanche gain and field emitter array (FEA) readout is being investigated. It is referred to as scintillator avalanche photoconductor with high resolution emitter readout (SAPHIRE). The present work investigates the temporal performance, i.e., lag, of SAPHIRE. METHODS: Since the temporal performance of the x-ray detection materials, i.e., the structured scintillator and avalanche amorphous selenium (a Se) photoconductor, has been studied previously, the investigation is focused on lag due to the FEA readout method. The principle of FEA readout is similar to that of scanning electron beam readout used in camera tubes, where the dominant source of lag is the energy spread of electrons. Since the principles of emission and beam focusing methods for FEA are different from thermionic emission used in camera tubes, its electron beam energy spread and hence lag is expected to be different. In the present work, the energy spread of the electrons emitted from a FEA was investigated theoretically by analyzing different contributing factors due to the FEA design and operations: The inherent energy spread of field emission, the FEA driving pulse delay, and the angular distribution of emitted electrons. The electron energy spread determined the beam acceptance characteristic curve of the photoconductive target, i.e., the accepted beam current (I(a)) as a function of target potential (V(t)), from which lag could be calculated numerically. Lag calculation was performed using FEA parameters of two prototype HARP-FEA image sensors, and the results were compared with experimental measurements. Strategies for reducing lag in SAPHIRE were proposed and analyzed. RESULTS: The theoretical analysis shows that the dominant factor for lag is the angular distribution of electrons emitted from the FEA. The first frame lags for two prototype sensors with 4 and 25 microm HARP layer thicknesses were 62.1% and 9.1%, respectively. A lag clearance procedure can be implemented by turning on all the FEA pixels simultaneously between subsequent frames without negative impact of readout speed. For large-area SAPHIRE, the bias electrode for the HARP needs to be divided into strips to allow parallel readout. With typical cardiac detector parameters, SAPHIRE with 128 parallel strips can provide real-time readout (30 frames/s) with first frame lag of -4%. CONCLUSIONS: The investigation of lag in SAPHIRE shows that the angular distribution of emitted electrons from FEA can result in substantial lag if the readout was performed pixel by pixel. Effective strategies for reducing lag include dividing the bias electrode into multiple strips to allow parallel readout and the incorporation of rapid charge clearance procedure between subsequent frames or rows. PMID- 19810478 TI - Spatial analysis of magnetic resonance T1rho and T2 relaxation times improves classification between subjects with and without osteoarthritis. AB - PURPOSE: Studies have shown that functional analysis of knee cartilage based on magnetic resonance (MR) relaxation times is a valuable tool in the understanding of osteoarthritis (OA). In this work, the regional spatial distribution of knee cartilage T1rho, and T2 relaxation times based on texture and laminar analyses was studied to investigate if they provide additional insight compared to global mean values in the study of OA. METHODS: Knee cartilage of 36 subjects, 19 healthy controls and 17 with mild OA, was divided into 16 compartments. T1rho and T2 relaxation times were studied with first order statistics, eight texture parameters with four different orientations using gray-level co-occurrence matrices and by subdividing each compartment into two different layers: Deep and superficial. Receiver operating characteristic curve analysis was performed to evaluate the potential of each technique to correctly classify the populations. RESULTS: Although the deep and superficial cartilage layers had in general significantly different T1rho and T2 relaxation times, they performed similarly in terms of subject discrimination. The subdivision of lateral and medial femoral compartments into weight-bearing and non-weight-bearing regions did not improve discrimination. Also it was found that the most sensitive region was the patella and that T1rho discriminated better than T2. The most important finding was that with respect to global mean values, laminar and texture analyses improved subject discrimination. CONCLUSIONS: Results of this study suggest that spatially assessing MR images of the knee cartilage relaxation times using laminar and texture analyses could lead to better and probably earlier identification of cartilage matrix abnormalities in subjects with OA. PMID- 19810479 TI - The M. D. Anderson proton therapy system. AB - PURPOSE: The purpose of this study is to describe the University of Texas M. D. Anderson proton therapy system (PTC-H) including the accelerator, beam transport, and treatment delivery systems, the functionality and clinical parameters for passive scattering and pencil beam scanning treatment modes, and the results of acceptance tests. METHODS: The PTC-H has a synchrotron (70-250 MeV) and four treatment rooms. An overall control system manages the treatment, physics, and service modes of operation. An independent safety system ensures the safety of patients, staff, and equipment. Three treatment rooms have isocentric gantries and one room has two fixed horizontal beamlines, which include a large-field treatment nozzle, used primarily for prostate treatments, and a small-field treatment nozzle for ocular treatments. Two gantry treatment rooms and the fixed beam treatment room have passive scattering nozzles. The third gantry has a pencil beam scanning nozzle for the delivery of intensity modulated proton treatments (IMPT) and single field uniform dose (SFUD) treatments. The PTC-H also has an experimental room with a fixed horizontal beamline and a passive scattering nozzle. The equipment described above was provided by Hitachi, Ltd. Treatment planning is performed using the Eclipse system from Varian Medical Systems and data management is handled by the MOSAIQ system from IMPAC Medical Systems, Inc. The large-field passive scattering nozzles use double scattering systems in which the first scatterers are physically integrated with the range modulation wheels. The proton beam is gated on the rotating range modulation wheels at gating angles designed to produce spread-out-Bragg peaks ranging in size from 2 to 16 g/cm2. Field sizes of up to 25 x 25 cm2 can be achieved with the double scattering system. The IMPT delivery technique is discrete spot scanning, which has a maximum field size of 30 x 30 cm2. Depth scanning is achieved by changing the energy extracted from the synchrotron (energy can be changed pulse to pulse). The PTC-H is fully integrated with DICOM-RT ION interfaces for imaging, treatment planning, data management, and treatment control functions. RESULTS: The proton therapy system passed all acceptance tests for both passive scattering and pencil beam scanning. Treatments with passive scattering began in May 2006 and treatments with the scanning system began in May 2008. The PTC-H was the first commercial system to demonstrate capabilities for IMPT treatments and the first in the United States to treat using SFUD techniques. The facility has been in clinical operation since May 2006 with up time of approximately 98%. CONCLUSIONS: As with most projects for which a considerable amount of new technology is developed and which have duration spanning several years, at project completion it was determined that several upgrades would improve the overall system performance. Some possible upgrades are discussed. Overall, the system has been very robust, accurate, reproducible, and reliable. The authors found the pencil beam scanning system to be particularly satisfactory; prostate treatments can be delivered on the scanning nozzle in less time than is required on the passive scattering nozzle. PMID- 19810480 TI - On the speckle-free nature of photoacoustic tomography. AB - PURPOSE: A long-standing conundrum is why photoacoustic tomography (PAT) possesses the unique ability to produce images devoid of speckle artifacts while all other coherent imaging technologies do not. METHODS: In this paper, we explain the inherent mechanism that suppresses speckle in PAT, and the analysis was validated by simulations based on an experimental PAT system. RESULTS: We found that the speckle-free feature of PAT results directly from the optical absorption contrast. CONCLUSIONS: All optical absorbers expand on laser excitation, and therefore all initial photoacoustic pressure rises are positive, which engenders strong correlations among the photoacoustic waves from the absorbers. As a result, prominent boundaries always build up in photoacoustic images and suppress the interior speckle. PMID- 19810481 TI - An effective correction algorithm for off-axis portal dosimetry errors. AB - Portal dosimetric images acquired for IMRT pretreatment verification show dose errors of up to 15% near the detector edges as compared to dose predictions calculated by a treatment planning system for these off-axis regions. A method is proposed to account for these off-axis effects by precisely correcting the off axis output factors, which calibrate the imager for absolute dose. Using this method, agreement between the predicted and the measured doses improves by up to 15% for fields near the detector edges, resulting in passing rate improvements of as much as 60% for gamma evaluation of 3 mm, 3% within the collimator jaws. PMID- 19810482 TI - Accelerated ray tracing for radiotherapy dose calculations on a GPU. AB - PURPOSE: The graphical processing unit (GPU) on modern graphics cards offers the possibility of accelerating arithmetically intensive tasks. By splitting the work into a large number of independent jobs, order-of-magnitude speedups are reported. In this article, the possible speedup of PLATO's ray tracing algorithm for dose calculations using a GPU is investigated. METHODS: A GPU version of the ray tracing algorithm was implemented using NVIDIA's CUDA, which extends the standard C language with functionality to program graphics cards. The developed algorithm was compared based on the accuracy and speed to a multithreaded version of the PLATO ray tracing algorithm. This comparison was performed for three test geometries, a phantom and two radiotherapy planning CT datasets (a pelvic and a head-and-neck case). For each geometry, four different source positions were evaluated. In addition to this, for the head-and-neck case also a vertex field was evaluated. RESULTS: The GPU algorithm was proven to be more accurate than the PLATO algorithm by elimination of the look-up table for z indices that introduces discretization errors in the reference algorithm. Speedups for ray tracing were found to be in the range of 2.1-10.1, relative to the multithreaded PLATO algorithm running four threads. For dose calculations the speedup measured was in the range of 1.5-6.2. For the speedup of both the ray tracing and the dose calculation, a strong dependency on the tested geometry was found. This dependency is related to the fraction of air within the patient's bounding box resulting in idle threads. CONCLUSIONS: With the use of a GPU, ray tracing for dose calculations can be performed accurately in considerably less time. Ray tracing was accelerated, on average, with a factor of 6 for the evaluated cases. Dose calculation for a single beam can typically be carried out in 0.6-0.9 s for clinically realistic datasets. These findings can be used in conventional planning to enable (nearly) real-time dose calculations. Also the importance for treatment optimization techniques is evident. PMID- 19810483 TI - Effect of prolonged stimulation on cerebral hemodynamic: a time-resolved fNIRS study. AB - PURPOSE: Sustained attention is one of the most important cognitive abilities for the management of everyday life, but it is often studied only at the behavioral level, while functional correlates are scarcely investigated. In this article, the authors address the topic of characterizing the dynamics of cerebral metabolism in the prefrontal cortex during a task of prolonged attention. METHOD: By means of multichannel time-resolved functional near-infrared spectroscopy and generalized linear model based data processing, the authors measured the hemodynamic response of the prefrontal cortex from 19 healthy subjects to a shortened version of a sustained attention task (Conners' Continuous Performance Test), lasting for 10 min. RESULTS: The task elicited significant brain activation, which did not remain constant for the entire task, but showed a drop not correlated with performance decay 4 min after the beginning of the task. Furthermore, oxygenated hemoglobin showed an increasing trend also during the first phase of the recovery, just after the end of the task. CONCLUSION: The results indicate a nontrivial dynamics of neural activation, habituation processes, and hemodynamic/metabolic coupling. These results encourage further studies about continuous stimulation of cognitive functions on both healthy and pathological subjects. PMID- 19810484 TI - Verification and source-position error analysis of film reconstruction techniques used in the brachytherapy planning systems. AB - A method was presented that employs standard linac QA tools to verify the accuracy of film reconstruction algorithms used in the brachytherapy planning system. Verification of reconstruction techniques is important as suggested in the ESTRO booklet 8: "The institution should verify the full process of any reconstruction technique employed clinically." Error modeling was also performed to analyze seed-position errors. The "isocentric beam checker" device was used in this work. It has a two-dimensional array of steel balls embedded on its surface. The checker was placed on the simulator couch with its center ball coincident with the simulator isocenter, and one axis of its cross marks parallel to the axis of gantry rotation. The gantry of the simulator was rotated to make the checker behave like a three-dimensional array of balls. Three algorithms used in the ABACUS treatment planning system: orthogonal film, 2-films-with-variable angle, and 3-films-with-variable-angle were tested. After exposing and digitizing the films, the position of each steel ball on the checker was reconstructed and compared to its true position, which can be accurately calculated. The results showed that the error is dependent on the object-isocenter distance, but not the magnification of the object. The averaged errors were less than 1 mm within the tolerance level defined by Roue et al. ["The EQUAL-ESTRO audit on geometric reconstruction techniques in brachytherapy," Radiother. Oncol. 78, 78-83 (2006)]. However, according to the error modeling, the theoretical error would be greater than 2 mm if the objects were located more than 20 cm away from the isocenter with a 0.5 degrees reading error of the gantry and collimator angles. Thus, in addition to carefully performing the QA of the gantry and collimator angle indicators, it is suggested that the patient, together with the applicators or seeds inside, should be placed close to the isocenter as much as possible. This method could be used to test the reconstruction techniques of any planning system, and the most suitable one can be chosen for clinical use. PMID- 19810485 TI - Uncertainties in effective dose estimates of adult CT head scans: the effect of head size. AB - PURPOSE: This study is an extension of a previous study where the uncertainties in effective dose estimates from adult CT head scans were calculated using four CT effective dose estimation methods, three of which were computer programs (CT EXPO, CTDOSIMETRY, and IMPACTDOSE) and one that involved the dose length product (DLP). However, that study did not include the uncertainty contribution due to variations in head sizes. METHODS: The uncertainties due to head size variations were estimated by first using the computer program data to calculate doses to small and large heads. These doses were then compared with doses calculated for the phantom heads used by the computer programs. An uncertainty was then assigned based on the difference between the small and large head doses and the doses of the phantom heads. RESULTS: The uncertainties due to head size variations alone were found to be between 4% and 26% depending on the method used and the patient gender. When these uncertainties were included with the results of the previous study, the overall uncertainties in effective dose estimates (stated at the 95% confidence interval) were 20%-31% (CT-EXPO), 15%-30% (CTDOSIMETRY), 20%-36% (IMPACTDOSE), and 31%-40% (DLP). CONCLUSIONS: For the computer programs, the lower overall uncertainties were still achieved when measured values of CT dose index were used rather than tabulated values. For DLP dose estimates, head size variations made the largest (for males) and second largest (for females) contributions to effective dose uncertainty. An improvement in the uncertainty of the DLP method dose estimates will be achieved if head size variation can be taken into account. PMID- 19810486 TI - Monte Carlo simulations of the count rate performance of a clinical whole-body MR/PET scanner. AB - The combination of MR and PET scanners can provide a powerful tool for clinical diagnosis and investigation. Among the existing approaches, the most challenging is that of complete hardware integration of both scanners. Such an integrated tomograph would allow simultaneous acquisition of both modalities, which could help solve issues such as cardiac and respiratory motion. Full integration imposes restrictions on the design of the PET part, such as detector configuration and maximum ring diameter. Furthermore, MR components surrounding the PET detector ring may cause gamma ray interactions, thus affecting PET performance. The purpose of this article is to assess the performance of a hypothetical whole-body integrated MR/PET scanner using Monte Carlo simulation techniques and compare it to state-of-the-art PET/CT devices used in clinical routine. The Monte Carlo simulation toolkit used for this study is the GEANT4 application for emission tomography. A hypothetical whole-body MR/PET tomograph fully integrated at hardware level and positioned between gradient and local coils of the MR scanner has been modeled. The NEMA 2-2001 protocol has been used to configure the simulations in order to measure sensitivity, scatter fraction, count losses, and random detections. Global sensitivity values as a function of the lower-level discriminator (LLD) energy are provided for time resolutions of 5 and 2.25 ns. In addition, the scatter fraction of the system is studied as a function of the LLD for energy resolution values of 10%, 15%, and 20%. Finally, true, scatter, random, and noise equivalent count rate curves as a function of activity concentration are given for dead-time values of 136, 432, and 1150 ns and for time resolution values of 2.25 and 5 ns. The influence on the count rate performance of the integrated PET scanner of the new geometry and interfering MR elements has been measured. The results show that the interference of the MR components has a much lower impact than the reduction in the detector ring diameter. Due to the larger solid angle coverage, the sensitivity is higher than that measured for a clinical PET/CT system (6200-10 900 cps/MBq at the center of the scanner) but not enough to compensate the degradation of the noise equivalent count rate due to increased scatter detection. The simulations prove the viability of an integrated MR/PET system and suggest that priority has to be given to either the improvement of the temporal resolution or the correction of triple coincidences if competitive performance is to be achieved. PMID- 19810487 TI - In vivo verification of proton beam path by using post-treatment PET/CT imaging. AB - PURPOSE: The purpose of this study is to establish the in vivo verification of proton beam path by using proton-activated positron emission distributions. METHODS: A total of 50 PET/CT imaging studies were performed on ten prostate cancer patients immediately after daily proton therapy treatment through a single lateral portal. The PET/CT and planning CT were registered by matching the pelvic bones, and the beam path of delivered protons was defined in vivo by the positron emission distribution seen only within the pelvic bones, referred to as the PET defined beam path. Because of the patient position correction at each fraction, the marker-defined beam path, determined by the centroid of implanted markers seen in the posttreatment (post-Tx) CT, is used for the planned beam path. The angular variation and discordance between the PET- and marker-defined paths were derived to investigate the intrafraction prostate motion. For studies with large discordance, the relative location between the centroid and pelvic bones seen in the post-Tx CT was examined. The PET/CT studies are categorized for distinguishing the prostate motion that occurred before or after beam delivery. The post-PET CT was acquired after PET imaging to investigate prostate motion due to physiological changes during the extended PET acquisition. RESULTS: The less than 2 degrees of angular variation indicates that the patient roll was minimal within the immobilization device. Thirty of the 50 studies with small discordance, referred as good cases, show a consistent alignment between the field edges and the positron emission distributions from the entrance to the distal edge. For those good cases, average displacements are 0.6 and 1.3 mm along the anterior-posterior (D(AP)) and superior-inferior (D(SI)) directions, respectively, with 1.6 mm standard deviations in both directions. For the remaining 20 studies demonstrating a large discordance (more than 6 mm in either D(AP) or D(SI)), 13 studies, referred as motion-after-Tx cases, also show large misalignment between the field edge and the positron emission distribution in lipomatous tissues around the prostate. These motion-after-Tx cases correspond to patients with large changes in volume of rectal gas between the post-Tx and the post-PET CTs. The standard deviations for D(AP) and D(SI) are 5.0 and 3.0 mm, respectively, for these motion-after-Tx cases. The final seven studies, referred to as position-error cases, which had a large discordance but no misalignment, were found to have deviations of 4.6 and 3.6 mm in D(AP) and D(SI), respectively. The position-error cases correspond to a large discrepancy on the relative location between the centroid and pelvic bones seen in post-Tx CT and recorded x ray radiographs. CONCLUSIONS: Systematic analyses of proton-activated positron emission distributions provide patient-specific information on prostate motion (sigmaM) and patient position variability (sigmap) during daily proton beam delivery. The less than 2 mm of displacement variations in the good cases indicates that population-based values of sigmap and sigmaM, used in margin algorithms for treatment planning at the authors' institution are valid for the majority of cases. However, a small fraction of PET/CT studies (approximately 14%) with -4 mm displacement variations may require different margins. Such data are useful in establishing patient-specific planning target volume margins. PMID- 19810488 TI - Monte Carlo model for a prototype CT-compatible, anatomically adaptive, shielded intracavitary brachytherapy applicator for the treatment of cervical cancer. AB - PURPOSE: Current, clinically applicable intracavitary brachytherapy applicators that utilize shielded ovoids contain a pair of tungsten-alloy shields which serve to reduce dose delivered to the rectum and bladder during source afterloading. After applicator insertion, these fixed shields are not necessarily positioned to provide optimal shielding of these critical structures due to variations in patient anatomies. The authors present a dosimetric evaluation of a novel prototype intracavitary brachytherapy ovoid [anatomically adaptive applicator (A3)], featuring a single shield whose position can be adjusted with two degrees of freedom: Rotation about and translation along the long axis of the ovoid. METHODS: The dosimetry of the device for a HDR 192Ir was characterized using radiochromic film measurements for various shield orientations. A MCNPX Monte Carlo model was developed of the prototype ovoid and integrated with a previously validated model of a v2 mHDR 192Ir source (Nucletron Co.). The model was validated for three distinct shield orientations using film measurements. RESULTS: For the most complex case, 91% of the absolute simulated and measured dose points agreed within 2% or 2 mm and 96% agreed within 10% or 2 mm. CONCLUSIONS: Validation of the Monte Carlo model facilitates future investigations into any dosimetric advantages the use of the A3 may have over the current state of art with respect to optimization and customization of dose delivery as a function of patient anatomical geometries. PMID- 19810489 TI - Monte Carlo simulation and patient dosimetry for a kilovoltage cone-beam CT unit. AB - PURPOSE: The purpose of this work is to characterize the x-ray volume imager (XVI), the cone-beam computed tomography (CBCT) unit mounted on the Elekta Synergy linac, with F1 bowtie filter and to calculate the three-dimensional dose delivered to patients using volumetric acquisition. METHODS: The XVI is modeled in detail using a new Monte Carlo (MC) code, BEAMPP, under development at the National Research Council Canada. In this investigation, a new component module is developed to accurately model the unit's bowtie filter used in conjunction with the available beam collimators at the clinical energy of 120 kV. The modeling is compared against percentage depth dose (PDD) and profile measurements. Kilovoltage radiation beams' phase space files are also analyzed. The authors also describe a method for the absolute dose calibration of the MC model of the CBCT unit when used in a clinical volumetric acquisition mode. Finally, they calculate three-dimensional patient dose from CBCT image acquisition in three clinical cases of interest: Pelvis, lung, and head and neck. RESULTS: The agreement between measurement and MC is shown to be very good: Within +/- 2% for the PDD and within +/- 3.5% inside the radiation field for all the collimators with the F1 bowtie filter. A full account of the absolute calibration method is given and dose calculation is validated against ion chamber measurements in different locations of a plastic phantom. Calculations and experiments agree within +/- 2% or better in both at the center and the periphery of the phantom, with worst agreement of 4.5% at the surface of the phantom and for one specific combination of collimator and filter. Patient dose from CBCT scan reveals that dose to tissue is between 2 and 2.5 cGy for a pelvis or a lung full acquisition. For H&N dose to tissue is 5 cGy, with the unit presets used in this work. Dose to bony structures can be two to three times higher than dose to tissue. CONCLUSIONS: The XVI CBCT unit has been fully modeled including the F1 bowtie filter. Absolute dose distribution from the unit has been successfully validated. Full MC patient dose calculation has shown that the three-dimensional dose distribution from CBCT is complex. Patient dose from CBCT exposure cannot be completely accounted for by using a numerical factor as an estimate of the dose at the center of the body. Furthermore, additional dose to bone should be taken into account when adopting any IGRT strategy and weighed vs the unquestionable benefits of the technique in order to optimize treatment. Full three-dimensional dose calculation is recommended if patient dose from CBCT is to be integrated in any adaptive planning strategy. PMID- 19810490 TI - Determination of the quality index (Q) for photon beams at arbitrary field sizes. AB - PURPOSE: A commonly used beam quality index (Q) for high-energy photon beams is the tissue phantom ratio (TPR20,10) for a square field of 10 x 10 cm2 and SDD of 100 cm. On some specialized radiotherapy treatment equipment such a reference collimator setting is not achievable. Likewise a flat beam profile, not explicitly required in dosimetry protocols, but certainly influences the measurement of Q, is not always produced. In this work, a method was developed in order to determine Q at any field size, especially for small and nonflattened beams. METHODS: An analytical relationship was derived between TPR20,10 for arbitrary field sizes and Q [the TPR20,10 (10 x 10 cm2)] as quality index. The proposed model equation was fitted to the measured and published data in order to achieve three general fit parameters. The procedure was then tested with published data from TomoTherapy and CyperKnife treatment devices. RESULTS: For standard flattened photon fields, the uncertainty in Q measured at any field size using the parameters derived from this study is better than 1%. In flattening filter free beams, the proposed procedure results in a reliable Q for any field size setting. CONCLUSIONS: A method is introduced and successfully tested in order to measure the beam quality under nonstandard conditions. It can be used, e.g., to get energy dependent correction factors as tabulated in dosimetry codes of practice even if standard conditions are not adjustable. PMID- 19810491 TI - Accuracy of Spencer-Attix cavity theory and calculations of fluence correction factors for the air kerma formalism. AB - EGSnrc calculations of ion chamber response and Spencer-Attix (SA) restricted stopping-power ratios are used to test the assumptions of the SA cavity theory and to assess the accuracy of this theory as it applies to the air kerma formalism for 60Co beams. Consistent with previous reports, the EGSnrc calculations show that the SA cavity theory, as it is normally applied, requires a correction for the perturbation of the charged particle fluence (K(fl)) by the presence of the cavity. The need for K(fl) corrections arises from the fact that the standard prescription for choosing the low-energy threshold delta in the SA restricted stopping-power ratio consistently underestimates the values of delta needed if no perturbation to the fluence is assumed. The use of fluence corrections can be avoided by appropriately choosing delta, but it is not clear how delta can be calculated from first principles. Values of delta required to avoid K(fl) corrections were found to be consistently higher than delta values obtained using the conventional approach and are also observed to be dependent on the composition of the wall in addition to the cavity size. Values of K(fl) have been calculated for many of the graphite-walled ion chambers used by the national metrology institutes around the world and found to be within 0.04% of unity in all cases, with an uncertainty of about 0.02%. PMID- 19810492 TI - A real-time prostate cancer detection technique using needle insertion force and patient-specific criteria during percutaneous intervention. AB - In this article, the authors present a novel real-time cancer detection technique by using needle insertion forces in conjunction with patient-specific criteria during percutaneous interventions. Needle insertion experiments and pathological analysis were performed for developing a computer-aided detection (CAD) model. Backward stepwise regression method was performed to identify the statistically significant patient-specific factors. A baseline force model was then developed using these significant factors. The threshold force model that estimated the lower bound of the cancerous tissue forces was formulated by adding an adjustable classifier to the baseline force model. Tradeoff between sensitivity and specificity was obtained by varying the threshold value of the classifier, from which the receiver-operating characteristic (ROC) curve was generated. Sequential quadratic programming was used to optimize the CAD model by maximizing the area under the ROC curve (AUC) using a set of model-training patient data. When the CAD model was evaluated using an independent set of model-validation patient data, an AUC of 0.90 was achieved. The feasibility of cancer detection in real time during percutaneous interventions was established. PMID- 19810493 TI - A general analytical solution to the geometrical problem of field matching in radiotherapy. AB - PURPOSE: Several authors studied the problem of geometrical matching of fields produced by medical linear accelerators. However, a general solution has yet to be published. Currently available solutions are based on parallelism arguments. This study provides a general solution, considering not only parallelism but also field sizes. METHODS: A fixed field with arbitrary field size, gantry, collimator, and couch angle is considered, and another field with a fixed gantry angle is matched to it. A single reference system attached to the treatment couch is used, and two approaches are followed. In the first approach, fixed field sizes are assumed and parallelism of the adjacent field-side planes is imposed. In the second approach, fixed isocenter positions are considered and both parallelism and coincidence between field-side planes are required. RESULTS: When fixed field sizes are assumed, rotation angles are obtained; however, the isocenters may need to be shifted to make side planes coincident and therefore achieve a proper match. When fixed isocenter positions are considered, solutions for all parameters, including the field size, are obtained and an exact geometrical match is achieved. CONCLUSIONS: General expressions to the field matching problem are found for the two approaches followed, fixed field sizes, and fixed isocenter positions. These results can be applied to any treatment technique and can easily be implemented in modern treatment planning systems. PMID- 19810494 TI - Task Group 142 report: quality assurance of medical accelerators. AB - The task group (TG) for quality assurance of medical accelerators was constituted by the American Association of Physicists in Medicine's Science Council under the direction of the Radiation Therapy Committee and the Quality Assurance and Outcome Improvement Subcommittee. The task group (TG-142) had two main charges. First to update, as needed, recommendations of Table II of the AAPM TG-40 report on quality assurance and second, to add recommendations for asymmetric jaws, multileaf collimation (MLC), and dynamic/virtual wedges. The TG accomplished the update to TG-40, specifying new test and tolerances, and has added recommendations for not only the new ancillary delivery technologies but also for imaging devices that are part of the linear accelerator. The imaging devices include x-ray imaging, photon portal imaging, and cone-beam CT. The TG report was designed to account for the types of treatments delivered with the particular machine. For example, machines that are used for radiosurgery treatments or intensity-modulated radiotherapy (IMRT) require different tests and/or tolerances. There are specific recommendations for MLC quality assurance for machines performing IMRT. The report also gives recommendations as to action levels for the physicists to implement particular actions, whether they are inspection, scheduled action, or immediate and corrective action. The report is geared to be flexible for the physicist to customize the QA program depending on clinical utility. There are specific tables according to daily, monthly, and annual reviews, along with unique tables for wedge systems, MLC, and imaging checks. The report also gives specific recommendations regarding setup of a QA program by the physicist in regards to building a QA team, establishing procedures, training of personnel, documentation, and end-to-end system checks. The tabulated items of this report have been considerably expanded as compared with the original TG-40 report and the recommended tolerances accommodate differences in the intended use of the machine functionality (non-IMRT, IMRT, and stereotactic delivery). PMID- 19810495 TI - Mass preserving nonrigid registration of CT lung images using cubic B-spline. AB - The authors propose a nonrigid image registration approach to align two computed tomography (CT)-derived lung datasets acquired during breath-holds at two inspiratory levels when the image distortion between the two volumes is large. The goal is to derive a three-dimensional warping function that can be used in association with computational fluid dynamics studies. In contrast to the sum of squared intensity difference (SSD), a new similarity criterion, the sum of squared tissue volume difference (SSTVD), is introduced to take into account changes in reconstructed Hounsfield units (scaled attenuation coefficient, HU) with inflation. This new criterion aims to minimize the local tissue volume difference within the lungs between matched regions, thus preserving the tissue mass of the lungs if the tissue density is assumed to be relatively constant. The local tissue volume difference is contributed by two factors: Change in the regional volume due to the deformation and change in the fractional tissue content in a region due to inflation. The change in the regional volume is calculated from the Jacobian value derived from the warping function and the change in the fractional tissue content is estimated from reconstructed HU based on quantitative CT measures. A composite of multilevel B-spline is adopted to deform images and a sufficient condition is imposed to ensure a one-to-one mapping even for a registration pair with large volume difference. Parameters of the transformation model are optimized by a limited-memory quasi-Newton minimization approach in a multiresolution framework. To evaluate the effectiveness of the new similarity measure, the authors performed registrations for six lung volume pairs. Over 100 annotated landmarks located at vessel bifurcations were generated using a semiautomatic system. The results show that the SSTVD method yields smaller average landmark errors than the SSD method across all six registration pairs. PMID- 19810496 TI - X-band EPR imaging as a tool for gradient dose reconstruction in irradiated bones. AB - PURPOSE: Various tools are currently available for dose reconstruction in individuals after accidental exposure to ionizing radiation. Among the available biological analyses, Monte Carlo simulations, and biophysical methods, such as electron paramagnetic resonance (EPR), the latter has proved its usefulness for retrospective dosimetry. Although EPR spectroscopy is probably the most sensitive technique, it does not provide spatial dosimetric data. This information is, however, highly desirable when steep dose gradient irradiations are involved. The purpose of this work was to explore the possibilities of EPR imaging (EPRI) for spatial dose reconstruction in irradiated biological material. METHODS: X-band EPRI was used to reconstruct ex vivo the relative dose distribution in human bone samples and hydroxyapatite phantoms after irradiation with brachytherapy seeds or x rays. Three situations were investigated: Homogeneous, stepwise gradient, and continuous gradient irradiation. RESULTS: EPRI gave a faithful relative spin density distribution in bone samples and in hydroxyapatite phantoms. Measured dose ratios were in close agreement with the actual delivered dose ratios. EPRI was able to distinguish the dose gradients induced by two different sources (125I and 192Ir). However, the measured spatial resolution of the system was 1.9 mm and this appeared to be a limiting factor. The method could be improved by using new signal postprocessing strategies. CONCLUSIONS: This study demonstrates that EPRI can be used to assess the regional relative dose distribution in irradiated bone samples. The method is currently applicable to ex vivo measurements of small size samples with low variation in tissue density but is likely to be adapted for in vivo application using L-band EPRI. PMID- 19810497 TI - 4D in-beam positron emission tomography for verification of motion-compensated ion beam therapy. AB - PURPOSE: Clinically safe and effective treatment of intrafractionally moving targets with scanned ion beams requires dedicated delivery techniques such as beam tracking. Apart from treatment delivery, also appropriate methods for validation of the actual tumor irradiation are highly desirable, In this contribution the feasibility of four-dimensionally (space and time) resolved, motion-compensated in-beam positron emission tomography (4DibPET) was addressed in experimental studies with scanned carbon ion beams. METHODS: A polymethyl methracrylate block sinusoidally moving left-right in beam's eye view was used as target. Radiological depth changes were introduced by placing a stationary ramp shaped absorber proximal of the moving target. Treatment delivery was compensated for motion by beam tracking. Time-resolved, motion-correlated in-beam PET data acquisition was performed during beam delivery with tracking the moving target and prolonged after beam delivery first with the activated target still in motion and, finally, with the target at rest. Motion-compensated 4DibPET imaging was implemented and the results were compared to a stationary reference irradiation of the same treatment field. Data were used to determine feasibility of 4DibPET but also to evaluate offline in comparison to in-beam PET acquisition. RESULTS: 4D in-beam as well as offline PET imaging was found to be feasible and offers the possibility to verify the correct functioning of beam tracking. Motion compensation of the imaged beta(+)-activity distribution allows recovery of the volumetric extension of the delivered field for direct comparison with the reference stationary condition. Observed differences in terms of lateral field extension and penumbra in the direction of motion were typically less than 1 mm for both imaging strategies in comparison to the corresponding reference distributions. However, in-beam imaging retained a better spatial correlation of the measured activity with the delivered dose. CONCLUSIONS: 4DibPET is a feasible and promising method to validate treatment delivery of scanned ion beams to moving targets. Further investigations will focus on more complex geometries and treatment planning studies with clinical data. PMID- 19810498 TI - Matching extended-SSD electron beams to multileaf collimated photon beams in the treatment of head and neck cancer. AB - PURPOSE: Matching the penumbra of a 6 MeV electron beam to the penumbra of a 6 MV photon beam is a dose optimization challenge, especially when the electron beam is applied from an extended source-to-surface distance (SSD), as in the case of some head and neck treatments. Traditionally low melting point alloy blocks have been used to define the photon beam shielding over the spinal cord region. However, these are inherently time consuming to construct and employ in the clinical situation. Multileaf collimators (MLCs) provide a fast and reproducible shielding option but generate geometrically nonconformal approximations to the desired beam edge definition. The effects of substituting Cerrobend for the MLC shielding mode in the context of beam matching with extended-SSD electron beams are the subject of this investigation. METHODS: Relative dose beam data from a Varian EX 2100 linear accelerator were acquired in a water tank under the 6 MeV electron beam at both standard and extended-SSD and under the 6 MV photon beam defined by Cerrobend and a number of MLC stepping regimes. The effect of increasing the electron beam SSD on the beam penumbra was assessed. MLC stepping was also assessed in terms of the effects on both the mean photon beam penumbra and the intraleaf dose-profile nonuniformity relative to the MLC midleaf. Computational techniques were used to combine the beam data so as to simulate composite relative dosimetry in the water tank, allowing fine control of beam abutment gap variation. Idealized volumetric dosimetry was generated based on the percentage depth-dose data for the beam modes and the abutment geometries involved. Comparison was made between each composite dosimetry dataset and the relevant ideal dosimetry dataset by way of subtraction. RESULTS: Weighted dose difference volume histograms (DDVHs) were produced, and these, in turn, summed to provide an overall dosimetry score for each abutment and shielding type/angle combination. Increasing the electron beam SSD increased the penumbra width (defined as the lateral distance of the 80% and 20% isodose contours) by 8-10 mm at the depths of 10-20 mm. Mean photon beam penumbra width increased with increased MLC stepping, and the mean MLC penumbra was approximately 1.5 times greater than that across the corresponding Cerrobend shielding. Intraleaf dose discrepancy in the direction orthogonal to the beam edge also increased with MLC stepping. CONCLUSIONS: The weighted DDVH comparison techniques allowed the composite dosimetry resulting from the interplay of the abovementioned variables to be ranked. The MLC dosimetry ranked as good or better than that resulting from beam matching with Cerrobend for all except large field overlaps (-2.5 mm gap). The results for the linear-weighted DDVH comparison suggest that optimal MLC abutment dosimetry results from an optical surface gap of around 1 +/- 0.5 mm. Furthermore, this appears reasonably lenient to abutment gap variation, such as that arising from uncertainty in beam markup or other setup errors. PMID- 19810499 TI - Evaluation of high-energy brachytherapy source electronic disequilibrium and dose from emitted electrons. AB - PURPOSE: The region of electronic disequilibrium near photon-emitting brachytherapy sources of high-energy radionuclides (60Co, 137CS, 192Ir, and 169Yb) and contributions to total dose from emitted electrons were studied using the GEANT4 and PENELOPE Monte Carlo codes. METHODS: Hypothetical sources with active and capsule materials mimicking those of actual sources but with spherical shape were examined. Dose contributions due to source photons, x rays, and bremsstrahlung; source beta-, Auger electrons, and internal conversion electrons; and water collisional kerma were scored. To determine if conclusions obtained for electronic equilibrium conditions and electron dose contribution to total dose for the representative spherical sources could be applied to actual sources, the 192Ir mHDR-v2 source model (Nucletron B.V., Veenendaal, The Netherlands) was simulated for comparison to spherical source results and to published data. RESULTS: Electronic equilibrium within 1% is reached for 60Co, 137CS, 192Ir, and 169Yb at distances greater than 7, 3.5, 2, and 1 mm from the source center, respectively, in agreement with other published studies. At 1 mm from the source center, the electron contributions to total dose are 1.9% and 9.4% for 60Co and 192Ir, respectively. Electron emissions become important (i.e., > 0.5%) within 3.3 mm of 60Co and 1.7 mm of 192Ir sources, yet are negligible over all distances for 137Cs and 169Yb. Electronic equilibrium conditions along the transversal source axis for the mHDR-v2 source are comparable to those of the spherical sources while electron dose to total dose contribution are quite different. CONCLUSIONS: Electronic equilibrium conditions obtained for spherical sources could be generalized to actual sources while electron contribution to total dose depends strongly on source dimensions, material composition, and electron spectra. PMID- 19810500 TI - SPECT imaging of high energy isotopes and isotopes with high energy contaminants with rotating slat collimators. AB - PURPOSE: Quantitative SPECT imaging is limited by many factors, including penetration of high energy photons through the collimator. Even small peaks of high energy yield extensive contamination in the photopeak energy window. Rotating slat collimators typically offer a 40- to 50-fold increase in geometric detection efficiency compared to parallel hole collimators, while the amount of high-energy contamination is expected to remain the same. The aim of this study is to show that SPECT for isotopes with high energy peaks benefits from the use of a rotating slat collimator by significantly reducing the effect of high energy contamination. METHODS: Monte Carlo simulations were performed for I-123 and I 131. The simulation of a single point source in a scattering medium was used to investigate the history of each detected photon both for the rotating slat and the parallel hole collimator. Next, realistic simulations of an image quality phantom in combination with iterative image reconstruction enabled us to study the gain in image quality which can be expected for the rotating slat collimator. RESULTS: The results show that the rotating slat collimator is able to reduce the contribution of high energy photons from about 60% to about 8% of all the detected photons in the photopeak energy window. This results in a recovered contrast gain of about 25%. For I-131, there was a reduction in contamination from 65% to 49% when using a rotating slat instead of a parallel hole collimator. This resulted in an average hot contrast improvement of 12% and average cold contrast improvement of 8% for tomographic images. CONCLUSIONS: Rotating slat collimators have an additional advantage regarding image quality for isotopes with high energy emissions, especially if the contamination arises from photons with energies higher than the main emission energy, due to the relatively lower number of measured collimator-penetrating photons with respect to geometrically collimated photons. PMID- 19810501 TI - Evaluation of nonrigid registration models for interfraction dose accumulation in radiotherapy. AB - PURPOSE: Interfraction dose accumulation is necessary to evaluate the dose distribution of an entire course of treatment by adding up multiple dose distributions of different treatment fractions. This accumulation of dose distributions is not straightforward as changes in the patient anatomy may occur during treatment. For this purpose, the accuracy of nonrigid registration methods is assessed for dose accumulation based on the calculated deformations fields. METHODS: A phantom study using a deformable cubic silicon phantom with implanted markers and a cylindrical silicon phantom with MOSFET detectors has been performed. The phantoms were deformed and images were acquired using a cone-beam CT imager. Dose calculations were performed on these CT scans using the treatment planning system. Nonrigid CT-based registration was performed using two different methods, the Morphons and Demons. The resulting deformation field was applied on the dose distribution. For both phantoms, accuracy of the registered dose distribution was assessed. For the cylindrical phantom, also measured dose values in the deformed conditions were compared with the dose values of the registered dose distributions. Finally, interfraction dose accumulation for two treatment fractions of a patient with primary rectal cancer has been performed and evaluated using isodose lines and the dose volume histograms of the target volume and normal tissue. RESULTS: A significant decrease in the difference in marker or MOSFET position was observed after nonrigid registration methods (p < 0.001) for both phantoms and with both methods, as well as a significant decrease in the dose estimation error (p < 0.01 for the cubic phantom and p < 0.001 for the cylindrical) with both methods. Considering the whole data set at once, the difference between estimated and measured doses was also significantly decreased using registration (p < 0.001 for both methods). The patient case showed a slightly underdosed planning target volume and an overdosed bladder volume due to anatomical deformations. CONCLUSIONS: Dose accumulation using nonrigid registration methods is possible using repeated CT imaging. This opens possibilities for interfraction dose accumulation and adaptive radiotherapy to incorporate possible differences in dose delivered to the target volume and organs at risk due to anatomical deformations. PMID- 19810502 TI - Gamma knife output factor measurements using VIP polymer gel dosimetry. AB - PURPOSE: Water equivalent polymer gel dosimeters and magnetic resonance imaging were employed to measure the output factors of the two smallest treatment fields available in a Gamma Knife model C radiosurgery unit, those formed employing the 4 and 8 mm final collimator helmets. METHODS: Three samples of the VIP normoxic gel formulation were prepared and irradiated so that a single shot of the field whose output factor is to be measured and a single shot of the reference 18 mm field were delivered in each one. Emphasis is given to the development and benchmarking of a refined data processing methodology of reduced uncertainty that fully exploits the 3D dose distributions registered in the dosimeters. RESULTS: Polymer gel results for the output factor of the 8 mm collimator helmet are found to be in close agreement with the corresponding value recommended by the vendor (0.955 +/- 0.007 versus 0.956, respectively). For the 4 mm collimator helmet, however, polymer gel results suggest an output factor 3% lower than the value recommended by the vendor (0.841 +/- 0.009 versus 0.870, respectively). CONCLUSIONS: A comparison with corresponding measurements published in the literature indicates that output factor results of this work are in agreement with those obtained using dosimetric systems which, besides fine spatial resolution and lack of angular and dose rate dependence of the dosimeter's response, share with polymer gels the favorable characteristic of minimal radiation field perturbation. PMID- 19810503 TI - Spatial registration of temporally separated whole breast 3D ultrasound images. AB - The purpose of this study was to evaluate the potential for use of image volume based registration (IVBaR) to aid in measurement of changes in the tumor during chemotherapy of breast cancer. Successful IVBaR could aid in the detection of such changes in response to neoadjuvant chemotherapy and potentially be useful for routine breast cancer screening and diagnosis. IVBaR was employed in a new method of automated estimation of tumor volume in studies following the radiologist identification of the tumor region in the prechemotherapy scan. The authors have also introduced a new semiautomated method for validation of registration based on Doppler ultrasound (U.S.) signals that are independent of the grayscale signals used for registration. This Institutional Review Board approved study was conducted on 10 patients undergoing chemotherapy and 14 patients with a suspicious/unknown mass scheduled to undergo biopsy. Reasonably reproducible mammographic positioning and nearly whole breast U.S. imaging were achieved. The image volume was registered offline with a mutual information cost function and global interpolation based on a thin-plate spline using MIAMI FUSE software developed at the University of Michigan. The success and accuracy of registration of the three dimensional (3D) U.S. image volume were measured by means of mean registration error (MRE). IVBaR was successful with MRE of 4.3 +/- 1.7 mm in 9 out of 10 reproducibility automated breast ultrasound (ABU) studies and in 12 out of 17 ABU image pairs collected before, during, or after 115 +/- 14 days of chemotherapy. Semiautomated tumor volume estimation was performed on registered image volumes giving 86 +/- 8% mean accuracy compared to the radiologist hand-segmented tumor volume on seven cases. Doppler studies yielded fractional volume of color pixels in the region surrounding the lesion and its change with changing breast compression. The Doppler study of patients with detectable blood flow included five patients with suspicious masses and three undergoing chemotherapy. Spatial alignment of the 3D blood vessel data from the Doppler studies provided independent measures for the validation of registration. In 15 Doppler image volume pairs scanned with differing breast compression, the mean centerline separation value was 1.5 +/- 0.6 mm, while MRE based on a few identifiable structural points common to the two grayscale image volumes was 1.1 +/- 0.6 mm. Another measure, the overlap ratio of blood vessels, was shown to increase from 0.32 to 0.59 (+84%) with IVBaR for pairs at various compression levels. These results show that successful registration of ABU scans may be accomplished for comparison and integration of information. PMID- 19810504 TI - [Research on AlInGaN quaternary alloys as MQW barriers in GaN-based laser diodes]. AB - InGaN/GaN, InGaN/InGaN and InGaN/AlInGaN multi-quantum-well (MQW) laser diodes (LDs) were grown on (0001) sapphire substrate by metalorganic chemical vapor deposition (MOCVD). The GaN (0002) synchrotron X-ray diffraction (XRD), electroluminescence (EL) and optical power-current (L-I) measurement reveal that AlInGaN quaternary alloys as barriers in MQWs can improve the crystal quality, optical emission performance, threshold current and slope efficiency of the laser diode structure to a large extent compared with other barriers. The relevant mechanisms are that: 1. The Al component increases the barrier height of the MQWs so that more current carriers will be caught in. 2. The In component counteracts the strain in the MQWs that decreases the dislocations and defects, thereby the nonradiative recombination centers are decreased. 3. The In component decreases the piezoelectric electric field that makes the electrons and the holes recombine more easily. PMID- 19810505 TI - [Applications of terahertz spectroscopy and imaging techniques in food safety inspection]. AB - With the development of ultra-fast laser technology and the further understanding of the mechanism behind the interaction of terahertz radiation and materials, THz radiation is actively developed as a new-style technology for a wide range of applications. Currently THz spectroscopy and imaging techniques show great promise in security detection, medical diagnosis, wireless communication, pharmaceutical and many other areas. Because of the growing importance of food safety issues, the detection technology of food safety is also faced with great opportunities and challenges, while THz spectroscopy and imaging techniques provide a new solution to the problem. Compared to other technologies, THz radiation with higher signal-to-noise ratio and wide dynamic range can simultaneously obtain both frequency-domain and time-domain information of the sample which relates to the information of physical structure and chemical composition of materials. Above all THz radiation features unique advantages in food safety inspection. In the present paper, the brief concept of terahertz radiation was introduced, while the properties and technical methods of THz wave were also discussed. The most recent progress in THz technology used in food safety inspection was summarized. The prospect and restricted factors of this novel technology in food industry were also discussed. PMID- 19810506 TI - [A method for resolving spectra shift in the urban air quality monitoring system (DOAS)]. AB - In the urban air quality monitoring system, there is spectra shift which is caused by environment factors on the optical part (temperature and optic fiber position), or by the self-change of Xe-lamp. Relative spectra shift will occur if the shift of lamp-spectrum and air-spectrum is inconsistent which has direct influences on the accuracy of the measurement results. So the match of wavelength between lamp-spectrum and air-spectrum should be considered when we retrieve pollutants concentration measurement of trace gas in the atmosphere through DOAS method. Based on the study of the unique structures for Xe-lamp emitting spectrum, a method for the calibration of two signal spectra using Xe-lamp emitting peak and least square fitting is given. The results show that, the impact of spectrum shift can be reduced by this method for retrieving results. PMID- 19810507 TI - [Micro- and nano-structures on metal induced by femtosecond laser radiation]. AB - In the present study the authors performed surface treatment of stainless steel 65Mn (the thickness is 60 microm) by femtosecond laser (pulse duration 148 fs, wavelength 775 nm). The single-pulse threshold could be obtained directly to be about 0. 2 J x cm(-2). The authors found that the femtosecond laser produced a large number of micro-structures such as nano-pores and nano-protrusions. Then the authors discussed the influence of pulse power and the number of shots on the formed surface structures. The authors found that with the change in the power and the number of pulses, the period of multiple parallel grooved surface patterns remained unchanged, which is about on the sub-micron level. Finally the authors processed the array of holes and the lines with different speed and number of pulses. PMID- 19810508 TI - [Time-resolved photoluminescence of stimulated emission from ZnO nanoparticles]. AB - The detailed room-temperature stimulated emission including its optical characteristics from nanosized ZnO particles prepared by homogenous precipitation method was investigated by time-resolved spectroscopy both from the frequency domain and time domain. As the excitation power was increased, sharp lasing peaks with the full width at half maximum less than 0.5 nm, similar to the FP lasing mode resonator mode, emerged rapidly from the emission spectra. Additionally, a narrow emission line intensity increased rapidly with increasing the excitation intensity, which was due to an exciton-exciton collision recombination, and the threshold excitation intensity was 7.2 GW x cm(-2). Upon higher excitation power, the lasing mechanism switched to electron-hole plasma (EHP). The EHP emission in the case of a higher excitation intensity appeared at a lower energy side of the E-E emission and replaced completely the E-E emission at the higher intensity. The EHP emission was red-shifted when further increasing the excitation intensity, compared to the E-E emissions. It was demonstrated that the red-shift of the EHP peak was attributed to the relevant band gap renormalization effect in the electron-hole plasma regime. At the same time, the emission lifetime was drastically reduced. Time-resolved spectrum of P band suggested a Gaussian-like decay time with only a few tens of picoseconds, compared to 2 ps, which is the limit of streak camera time resolution. The dynamic processes of lasing behavior and characteristics of the lasing emission in ZnO nanoparticles could be valuable and provide the information on crystal quality, exciton and lasing action in ZnO. PMID- 19810509 TI - [Study of CO2 spectroscopic parameters at high temperature near 1.57 microm]. AB - Measurements strategies based on absorption spectroscopy techniques, especially the measurements in high temperature, require accurate values of important spectroscopic parameters of the probed species. Sometimes the parameters listed in widely used HITRAN and HITEMP2004 database are uncertain to some extent. In order to validate the spectroscopic parameters of 9 selected CO2 lines which should be used in combustion diagnosis, spectra of those lines were recorded in a high temperature experiment setup as a function of temperature (in the range of 300-800 K) and pressure (in the range of 9-450 torr) using a distributed feed back (DFB) diode laser. The recorded absorption spectra were fitted to Voigt profile. Line intensity, air-broadening coefficient and temperature exponent of each line were deduced from those data. Through comparison of experimental results and those listed in HITRAN and HITEMP2004 database, the discrepancies of most line intensities, air-broadening coefficients and their temperature exponents are less than 3%, 5% and 2% respectively. Those results show good consistency between the experimental data and that in HITRAN and HITEMP2004 database. The discrepancy in line intensities may be caused by the fitting of absorption spectra, the reading of thermocouple and pressure gage, uniformity of temperature in the heated cell, and uncertainty of the optical path. Those factors also cause the discrepancy in air-broadening coefficients and their temperature exponent. CO2 contained in air also introduces error in air broadening coefficients and their temperature exponent beside those factors. Though we have deducted them in data-processing, the little change of CO2 in partial region also exists. Those results will be helpful to the measurement of CO2 concentration in combustion diagnosis in the future. PMID- 19810510 TI - [Dual-wavelength Mie lidar observations of tropospheric aerosols]. AB - A new dual-wavelength Mie lidar (DWL) is introduced. The DWL can be used to monitor the optical properties of tropospheric aerosol at 532 and 1 064 nm wavelength and their spatial and temporal variations, and to research aerosol size distribution with altitude. This lidar adopted four channels to receive the far and near range backscattering signal at 532 and 1 064 nm wavelength respectively. In order to enhance the capability of daytime measurement, the system employed a narrow band interference filter to separate the main backscattering signal of lidar return, including Mie backscattering signal and Rayleigh backscattering signal from the total backscattering signal including non elastic scattering signal and solar spectrum, by cooperating with an iris to depress the majority of sky background noise. Overall structure and specifications of the lidar, as well as data processing method, were described. The lidar system has been operated in Hefei (117. 16 degrees E, 31.90 degrees N). The profile of extinction coefficient of tropospheric aerosol and its temporal spatial distribution were obtained. Angstrom exponent and optical depth of aerosol were also discussed. The observational results have shown that this lidar works well both during the day and at night and has the ability to measure the tropospheric aerosols and to manifest the temporal and spatial distributions of the aerosols with high precision. PMID- 19810512 TI - [MTF of line-array CCD on detection performance for hyperspectral imager]. AB - As an image-spectrum merging technology, hyperspectral imaging has been used in battlefield reconnaissance rapidly. According to the definition of MTF based on contrast, the theoretical model of the MTF of line-array CCD was presented. The effect of the relative position of linearray CCD and input signal on MTF was analyzed. The results indicate that at the frequency-division of Nyquist frequency, the relative position has a strong effect on the MTF, which becomes much stronger with frequency increase, while at other frequency, the effect of the relative position on MTF depends on the percent error zeta, e.g. the bigger the value of [zeta], the weaker the effect of the relative position on MTF. In the frequency range of [zeta] > 0.1%, the effect of the relative position on MTF can be ignored. The above conclusions have directive significance in the design of MTF measure system of hyperspectral imager. PMID- 19810511 TI - [Colon adenoma detection using Kubelka-Munk spectral function of DNA and protein bands]. AB - Differential diagnosis of human colon adenoma was studied using the Kubelka-Munk spectral function of the DNA and protein absorption bands at 260 and 280 nm in vitro. Diffuse reflectance spectra of tissue were measured using a spectrophotometer with an integrating sphere attachment. The results of measurement showed that for the spectral range from 590 to 1 064 nm pathological changes of colon epithelial tissues were induced so that there were significant differences in the averaged values of the Kubelka-Munk function f(r infinity) and logarithmic Kubelka-Munk function log [f(r infinity)] of the DNA absorption bands at 260 nm between normal and adenomatous colon epithelial tissues, and the differences were 218% (p < 0.05) and 68.5% (p < 0.05) respectively. Pathological changes of colon epithelial tissues were induced so that there were significant differences in the averaged values of the Kubelka-Munk function f(r infinity) and logarithmic Kubelka-Munk function log [f(r infinity)] of the protein absorption bands at 280 nm between normal and adenomatous colon epithelial tissues, and the differences were 208% (p < 0.05) and 59.0% (p < 0.05) respectively. Pathological changes of colon epithelial tissues were induced so that there were significant differences in the averaged values of the Kubelka-Munk function f(r infinity) and logarithmic Kubelka-Munk function log [f(r infinity)] of the beta-carotene absorption bands at 480 nm between normal and adenomatous colon epithelial tissues, and the differences were 41.7% (p < 0.05) and 32.9% (p < 0.05) respectively. Obviously, pathological changes of colon epithelial tissues were induced so that there were significant changes in the contents of the DNA, protein and beta-carotene of colon epithelial tissues. The conclusion can be applied to rapid, low-cost and noninvasive optical biopsy of colon adenoma, and provides a useful reference. PMID- 19810513 TI - [Synthesis of 1, 2, 5-triphenyl-pyrrole and effect of its aggregation degree on photoluminescence intensity]. AB - 1,2,5-triphenylpyrrole (TPP) was firstly prepared by the Schulte-Reisch reaction of 1,4-diphenylbuta-1,3-diyne with aniline catalyzed by copper chloride. Compared to solution reaction in DMF as solvent, the bulk reaction modified in this paper not only increased the yield and reduced the reaction temperature, but also shortend the reaction time. The pi-pi stacking interaction and the restriction of intramolecular rotation are were involved at the same time when TPP was aggregated in the THF-water mixtures. When the water volume fraction was under 60%, the PL intensity of TPP was independent on the water fraction in THF-water mixture. When the water fraction was added to 70%, which induced the non-tight aggregation of TPP, the strong pi-pi stacking interaction toned the nonradiative deactivation process and led to quenching the fluorescence of TPP; if the water fraction was further increased to 80%, which induced the tight aggregation of TPP, the restriction of intramolecular rotation was, however, preponderant over pi-pi stacking interaction. Thus the nonradiative channel was blocked and the photoluminescence intensity of TPP was enhanced. The compact degree of aggregation was influenced by acetonitrile solvent due to the charge transfer interaction between TPP and acetonitrile. The aggregation-induced emission enhancement of TPP in THF-water mixture disappeared in acetonitrile-water mixture. PMID- 19810514 TI - [Effect of Ce3+ implantation on photoluminescence intensity of Si nanocrystals embedded in superlattices]. AB - In the present paper, SiO/SiO2 superlattices samples were prepared on Si substrates by electron beam evaporation. The samples were annealed in nitrogen atmosphere at high temperature subsequently. And then, Ce3+ ions with a dose of 2.0 x 10(14) and 2.0 x 10(15) cm(-2) respectively were implanted into these samples with formed Si nanocrystals. The photoluminescence (PL) spectra showed that the PL intensities of samples with Ce3+ implanting dropped sharply compared with the samples without Ce3+ implanting. The PL intensity increased gradually with increasing re-annealing temperature, but dropped again when the temperature exceeded 600 degrees C. The PL intensity even could be higher than that of samples without Ce3+ implanting if only the dose of Ce3+ was 2.0 x 10(14) cm(-2). When the dose of Ce3+ was 2.0 x 10(15) cm(-2), the PL intensity couldn't exceed that of samples without Ce3+ implanting even when the re-annealing temperature was 600 degrees C. Further investigations showed that the varieties of the PL intensities were mainly dependent on the re-annealing temperature, which had the best point at 600 degrees C, and the dose of Ce3+ had the right value. Furthermore, the experiment results proved that there was energy transfer from Ce3+ to Si nanocrystals in this kind of structure. PMID- 19810515 TI - [Upconversion luminescence properties of ZBLAN : Nd3+, Tm3+, Yb3+]. AB - The ZBLAN : Nd3+, Tm3+, Yb3+ upconversion materials were prepared by high temperature solid method. The concentrations of Tm3+ and Yb3+ were 0.01% and 0.3% respectively. The concentration of Nd3+ changed in the range from 0.1% to 2%. The absorption spectrum of ZBLAN : Nd3+, Tm3+, Yb3 in all samples from 300 to 1 000 nm was measured at room temperature. The upconversion emission was observed when excited by 798 nm infrared light. The material samples emitted relatively strong multi-band (the red, blue and green color) visible light. Based on the experiment, the multi-band visible spectral lines were analyzed, and the energy level transition mechanism was given. The blue light results from the transition of 1G4 --> 3 H6 of Tm3+, the green light is from the transition of 2 H7/2 --> 4 I9/2 of Nd3+, and the red light originates from the transition of 2 H11/2 --> I9/2 of Nd3+. The results show that the upconversion mechanism includes excited state absorption, energy transfer, cross relaxation and so on. The energy transfer processes between Nd3+, Tm3+ and Yb3+ ions both forward and backward prove to be the origin of upconversion emissions. The effect of rare earth doping concentration on the energy upconversion efficiency was analyzed, according to the different upconversion light emitting intensity from samples with different Tm3+ mol concentrations. The upconversion luminescence increased with the Nd3+ concentration and got its peak at 1.5%. PMID- 19810516 TI - [Stray light and bandpass correction in the spectral measurement for light emitting diodes]. AB - The present paper introduces the stray light and bandpass correction methods for spectrometers. The line spread function of spectrometer is characterized by a He Ne laser. Assuming that the spectrometer is a wavelength invariable system, the stray light distribution matrix is constructed by the derived line spread functions. The stray light correction matrix is then derived by matrix conversion from the stray light distribution matrix. The measured signals of the spectrometer are finally multiplied by the stray light correction matrix to correct the stray light errors. The bandpass functions of the spectrometer are characterized in three different wavelength ranges, respectively. And then three groups of bandpass correction coefficients are calculated accordingly. The calculation is divided into several steps. Given the measurement results at the target wavelength position and the ones on the neighbor bandwidths, the bandpass correction results are obtained by weight averaging of them. The bandpass correction coefficients are used as the weights. The two correction methods are applied to a multi-channel fast spectrometer to measure LEDs of different color. The results show that the stray light and the bandpass errors can be corrected effectively. The chromaticity coordinates of the LEDs are corrected by (-0.003, 0.007) for the maximum. Furthermore, the method introduced in this paper can reduce the application cost, simplify the calculation under a reasonable precision, and make the application of the correction easier. PMID- 19810517 TI - [Study on Aconitum kusnezoffii Reichb. and its processed products by 2D-IR correlation spectroscopy]. AB - 2D-IR correlation spectroscopy and dynamic spectra tracked during mutative temperature process were used to do the research. The FTIR spectra of aconitum kusnezoffii Reichb. and its processed products are similar, the result by second derivative spectrum was analyzed, the absorption peaks at 1 745, 1 468 and 1 337 cm(-1) shifted to the low wave number, while the absorption peak at 1 657 cm(-1) shifted to the high wave number after being processed with acidophilus milk. Obvious differences are observed between 2D-IR spectra of them, active peaks of aconitum kusnezoffii Reichb. 1 650, 1 560 and 1 470 cm(-1) were recorded in the region of 1 300-800 cm(-1), and the strongest was at 1 560 cm(-1). Automatic peaks and cross peaks presented a symmetric distribution of 3 x 3, and all were positive correlation. Automatic peaks of processed aconitum kusnezoffii Reichb. in the region were in four areas, one was 1 220 and 1 200 cm(-1), the second was sharp peaks of 1 140 and 1 070 cm(-1), the third was wide peaks in the region of 1 000-900 cm(-1), and all the automatic peaks were positive correlation. Therefore, aconitum kusnezoffii Reichb. and its processed products can be identified intuitively by automatic and cross peaks of 2D-IR spectra, and the change laws of functional groups of them can be revealed. The method is rapid and exact, and can provide the means to analyze the structures' change laws of aconitum kusnezoffii Reichb. after being processed. PMID- 19810518 TI - [Identification of Cistanche deserticola from Boschniakla rossica and Cynomorium songaricum using FTIR and two-dimensional correlation IR spectroscopy]. AB - In the present paper, Fourier transform infrared spectroscopy (FTIR) combined with two-dimensional correlation spectroscopy (2D-IR) was used for the identification of Cistanche deserticola from its adulterants, Boschniakla rossica and Cynomorium songaricum. The results showed that these IR spectra showed their different macro-fingerprint feathers: the charactersistic peaks for Cistanche deserticola were located at -1 730 and 931 cm(-1). While absorption peaks at -1 510, 1 375 and 1 266 cm(-1) were only found in Boschniakla rossica. And the strongest peak of Cynomorium songaricum was at 1 614 cm(-1). Differences were more obvious in the second derivative IR spectra: the intensities of the four peaks of Cistanche deserticola at -1 453,1 336, 931, and 892 cm(-1) were strong, among which the peak at -931 cm(-1) was sharp and the strongest in intensity. There was the strongest peak at -1 509 cm(-1) for Boschniakla rossica. In addition, the shape of peaks at -1 633 and 1 161 cm(-1) was wider and the intensities were stronger. While the shape of peak at -1 682 cm(-1) was sharp, and also the intensity of peak at 1 605 cm(-1) was stronger. According to the features of the IR spectra of each kind, a quick and accurate method based on FTIR for the identification of Cistanche deserticola was established. PMID- 19810519 TI - [Effects of different managements on soil humic acid structural features in chestnut soil on typical Leymus chinensis steppe, Inner Mongolia, China]. AB - Grazing has a lot of effects on grassland soil properties. The objective of the present study is to investigate the effects of different managements on soil humic acids. Soil samples were collected from the surface soil of four adjacent plots ( I , II , III and IV) at a long-term experiment site of the Inner Mongolia Grassland Ecosystem Research Station (IMGERS), the Chinese Academy of Sciences. The structure features of humic acids were investigated by cross-polarization magic angle spinning 13C-nuclear magnetic resonance spectroscopy (CP/MAS 13C NMR), FTIR and elemental analysis. The results indicated that the humic acid extracted from the grazing plots were characterized by a higher degree of humification, including loss of polysaccharides, decrease in lignin content and increase in aromaticity. Compared with the humic substances from the grazing plots, the humic substances from fenced plots showed a lower degree of humification and were considerably more aliphatic in nature. PMID- 19810520 TI - [Research on systematic evolution of ginkgo biloba based on chemical composition of wood]. AB - In the present article, the authors mensurated the infrared spectra of 22 conifer and broadleaf trees such as ginkgo biloba L., cycas revoluta thunb., Populus tomentosa Carr and so on using FTIR, analyzed the difference of a number of absorb peaks in fingerprint area, the characteristic absorb peak position of cellulose and lignin of these trees and their alternation regulations of relative intensity, then discussed the evolutional issue of ginkgo biloba. The results show that ginkgo biloba is different from cycas in the infrared spectra of wood but similar to the conifer trees, which illustrates the ginkgo biloba. is more evolutional than cycas in the timber's chemical composition but with similar affiliation to conifer trees. The content of lignin in the ginkgo biloba wood is more than that in other conifer trees, which may be a original character of ginkgo biloba. Meanwhile, the ginkgo biloba has some syringyl lignins in addition to guaiacyl, and it has the evolutional tendency for broadleaf trees. PMID- 19810521 TI - [Temperature compensation for calibration model of apple fruit soluble solids contents by near infrared reflectance]. AB - The detection precision of soluble solids in apple fruit by near infrared reflectance (NIR) spectroscopy was affected by sample temperature. The NIR technique needs to be able to compensate for fruit temperature fluctuations. In the present study, it was observed that the sample temperature (2-42 degrees C) affects the NIR spectrum in a nonlinear way. The temperature model was built with R2 = 0.985, RMSEC = 1.88, and RMSEP = 2.32. When no precautions are taken, the error in the SSC reading may be as large as 2.55% degrees Brix. Two techniques were found well suited to control the accuracy of the calibration models for soluble solids with respect to temperature fluctuations, such as temperature variable--eliminating calibration model and global robust calibration model to cover the temperature range. And an improved genetic algorithms (GAs) was used to implement an automated variables selection procedure for use in building multivariate calibration models based on partial least squares regression (PLS). The two compensation methods were found to perform well with RMSEP1 = 0.72/0.69 and RMSEP2 = 0.74/0.68, respectively. This work proved that the compensation techniques could emend the temperature effect for NIR spectra and improve the precision of models for apple SSC by NIR. PMID- 19810522 TI - [Study on analysis of copy paper by Fourier transform infrared spectroscopy]. AB - A new method of fast identification of copy papers by Fourier transform infrared spectroscopy (FTIR) was developed. The kinds of filler and the cellulosic degree of crystallinity were analyzed by FTIR, and the ageing curves of cellulosic paper were studied with heating and ultraviolet light. The cellulosic degree of crystallinity was showed by the ratio of absorbance at 1 429 cm(-1) to that at 893 cm(-1), the standard deviation of different brands of copy papers was 0.010 7 0.016 0, and the standard deviation of the same brands of copy papers was 0.014 8. The kinds of filler and the cellulosic degree of crystallinity were different in copy papers from different brands of different manufacturing plants, different brands of same manufacturing plants and different manufacturing times of the same brands from the same manufacturing plants, and the curves of ageing were different with heating and ultraviolet light. The results of fast identification of copy papers by FTIR are satisfactory. PMID- 19810523 TI - [Study on relationship between on-the-go near-infrared spectroscopy and soil texture]. AB - Near infrared spectroscopy (NIRS) is a rapid, pioximal-sensed method that has proven useful in quantifying soil constituents mainly in laboratory. However, very little is known about how NIRS performs in a field setting by newly developed on-the-go NIRS measurements. The objective of the present study was to evaluate the relationship between on-the-go field NIRS measurements and soil texture in a glacial till soil. It was found that NIRS band combination based on difference, normalized difference and ratio could apparently improve the coefficient of relationship between NIRS and soil texture, and this might be a new and effective analytical procedure for field NIRS measurements. PMID- 19810524 TI - [Structural characteristics of humic acids from a long-term petroleum contaminated soil]. AB - Petroleum contamination in soil decreases with the increase in the distance of soil to the drilling well. Accordingly, an abandoned petroleum well which had been exploited for about twenty years in Songyuan city of Jilin Province, China, was selected to investigate the structural characteristics of soil humic acids (HAs) under different petroleum contamination levels. Surface (0-20 cm) soil samples were collected at 0.5, 1.5, 3.5, 5.5 and 7.5 m deep from well head, and the petroleum contents were respectively 153.3, 148.4, 129.2, 50.5 and 5.62 g x kg(-1). HAs were extracted with 0.1 mol x L(-) NaOH and 0.1 mol x L(-1) Na4 P2O7 and were characterized with elemental analysis, Fourier transformed infrared (FTIR) spectroscopy and solid-state 13C cross polarization magic angle spinning nuclear magnetic resonance (13C CPMAS NMR) spectroscopy. Results showed that the atomic C/H, O/C and (N+O) /C ratios of HAs increased from 0.74, 0.41 and 0.45 for 7.5 m to 0.80, 0.83 and 0.88 for 0.5 m, respectively. The relative intensity of the peaks assigned to aliphatic carbon (2 921, 2 851 and 1 454 cm(-1)) in the FTIR spectra gradually decreased with increasing contamination levels, while that of the peak assigned to aromatic C(1 600 cm(-1)) increased, and the calculated absorption intensity ratio of 2 921 to 1 600 cm(-1) (2 921/1 600) declined from 0.22 for 7.5 m to 0.11 for 0.5 m. The solid-state 13C NMR data suggested that the relative content of alkyl C(0-50 ppm ) decreased from 49.9% for 7.5m to 30.9% for 0.5 m, while that of O-alkyl C(50-110 ppm), aromatic C(110-160 ppm) and carboxyl C(160-190 ppm) increased respectively from 20.1%, 13.1% and 14.3% to 28.0%, 18.8% and 19.3%. These results showed substantial chemical, structural, and molecular differences among these HAs. The aliphaticity and hydrophobicity of HAs decreased while aromaticity and polarity increased with the increase in petroleum content. Namely, HAs tended to become aged in molecular structure. Therefore, it is imperative to renew and activate the aged HAs by adopting appropriate measures for the remediation of petroleum contaminated soil. PMID- 19810525 TI - [Study on Vis/NIR spectra detecting system for watermelons and quality predicting in motion]. AB - To make Vis/NIR diffuse transmittance technique applied to quality prediction for watermelon in motion, the dynamic spectra detecting system was rebuilt. Spectra detecting experiments were conducted and the effects of noises caused by motion on spectra were analyzed. Then the least--square filtering method and Norris differential filtering method were adopted to eliminate the effects of noise on spectra smoothing, and statistical models between the spectra and soluble solids content were developed using partial least square method. The performance of different models was assessed in terms of correlation coefficients (r) of validation set of samples, root mean square errors of calibration (RMSEC) and root mean square errors of prediction (RMSEP). Calibration and prediction results indicated that Norris differential method was an effective method to smooth spectra and improve calibration and prediction results, especially, with r of 0.895, RMSEC of 0.549, and RMSEP of 0.760 for the calibration and prediction result of the first derivative spectra. PMID- 19810526 TI - [Identification of varieties of cashmere by Vis/NIR spectroscopy technology based on PCA-SVM]. AB - One mixed algorithm was presented to discriminate cashmere varieties with principal component analysis (PCA) and support vector machine (SVM). Cashmere fiber has such characteristics as threadlike, softness, glossiness and high tensile strength. The quality characters and economic value of each breed of cashmere are very different. In order to safeguard the consumer's rights and guarantee the quality of cashmere product, quickly, efficiently and correctly identifying cashmere has significant meaning to the production and transaction of cashmere material. The present research adopts Vis/NIRS spectroscopy diffuse techniques to collect the spectral data of cashmere. The near infrared fingerprint of cashmere was acquired by principal component analysis (PCA), and support vector machine (SVM) methods were used to further identify the cashmere material. The result of PCA indicated that the score map made by the scores of PC1, PC2 and PC3 was used, and 10 principal components (PCs) were selected as the input of support vector machine (SVM) based on the reliabilities of PCs of 99.99%. One hundred cashmere samples were used for calibration and the remaining 75 cashmere samples were used for validation. A one-against-all multi-class SVM model was built, the capabilities of SVM with different kernel function were comparatively analyzed, and the result showed that SVM possessing with the Gaussian kernel function has the best identification capabilities with the accuracy of 100%. This research indicated that the data mining method of PCA-SVM has a good identification effect, and can work as a new method for rapid identification of cashmere material varieties. PMID- 19810527 TI - [FTIR analysis of products derived from wood liquefaction with 1-octanol]. AB - Solvolysis is one of the important processes of biomass liquefaction. To produce superior quality liquid biofuel from biomass under mild conditions, it is essential to exploit novel reactive liquid solvent. Furthermore, the evaluation of liquefaction efficiency is carried out mainly by the means of analysis of the products derived from biomass liquefaction. In the present study, liquefaction of poplar wood powder in acidified 1-octanol was investigated with a stainless steel autoclave. Residue, heavy oil and light oil were separated from the liquefaction products by extraction with acetone and n-hexane successively. FTIR analysis was carried out on these liquefaction compositions to illuminate the liquefaction regularities and mechanisms of cellulose, hemicellulose and lignin of wood. The results showed that liquefaction oils were complex mixture containing hydroxide, carbonyl, methoxyl, aromatic and aether. Liquefactions of cellulose and hemicellulose were easier than that of lignin. Cellulose and hemicellulose were converted to light oil, however, lignin was mainly converted to heavy oil. At 150 degrees C, lignin was depolymerized and degraded into micromolecular aromatic compounds, among which condensation reactions took place when reaction temperature increased. PMID- 19810528 TI - [Application of wavelet packet analysis in estimating soil parameters based on NIR spectra]. AB - Using the method of wavelet analysis, the NIR spectra of soil samples were decomposed and reconstructed, and higher precision PLS models were established to estimate soil parameter (TN, SOM). One hundred fifty soil samples were collected from a winter wheat field and the NIR spectra of all samples were measured. Firstly, experiment statistic features were analyzed aiming at all soil samples, and the system clustering was carried out for TN and SOM respectively. Then 50 new TN samples and their corresponding spectra, and 50 new SOM samples and their corresponding spectra were obtained. Secondly, the PLS models were established with these new samples based on their corresponding spectra. The models showed a certain amount of accuracy, but it was still not practical. Therefore, wavelet analysis of NIR spectra was tried. The wavelet packet decomposing by eight-level biorthogonal algorithm was carried out, and 256 nodes were gotten. The lowest approximation signal is corresponding to soil moisture and soil texture spectrum trend. The maximal detail signal is corresponding to the high-frequency turbulence caused by the soil particle size, precision of spectrometer, and other uncertainties. After reconstructing these two nodes and then removed from the original spectra, the characteristic spectra corresponding to each soil parameter were acquired. Finally, the PLS models were established for TN and SOM content respectively: for TN content, the calibration coefficient of the PLS model is 0.960, the validation coefficient is 0.920; and for SOM content, the calibration coefficient of the PLS model is 0.922, and the validation coefficient is 0.883. It was showed that the accuracy of each model was highly improved and the models were able to meet the needs of actual production. The research results conclude that wavelet analysis can eliminate or substantially reduce the factors outside the parameters. It can also remove the obstacles in establishing linear models of soil parameters, and it is feasible and potential method for the real-time estimation of soil parameters. PMID- 19810529 TI - [Application of probabilistic neural networks method to gastric endoscope samples diagnosis based on FTIR spectroscopy]. AB - In the present paper, probabilistic neural network method was applied to the classification of gastric endoscope samples based on FTIR spectroscopy for higher discrimination correctness than the conventional linear discriminant analysis algorithm. The probabilistic neural network (PNN) is a kind of radial basis network suitable for discriminant analysis. There are several advantages of PNN method: less time is needed to train the model, higher correctness could be achieved, global optimal solution could be obtained and so on. In this paper, PNN method was utilized to classify gastric endoscopic biopsies into healthy, gastritis, and malignancy. Firstly, principal component analysis was carried out for the pretreated sample spectra. Principal components analysis is a quantitatively rigorous method for achieving the simplification. The method generates a new set of variables, called principal components. Each principal component is a linear combination of the original variables. All the principal components are orthogonal to each other, so there is no redundant information. The principal components as a whole form an orthogonal basis for the space of the data. And then, the scores of principal components were selected as input to train the PNN model. Finally, PNN model was established. In this experiment, a total of 118 gastric endoscopic biopsies, including 35 cases of cancer, 64 cases of gastritis, and 19 healthy tissue samples, were obtained at the First Hospital of Xi'an Jiaotong University, China. Fifty nine samples were selected to establish the PNN classification model. The rest of the samples were used as the test set to valid the discriminant analysis model. The total discrimination correctness of normal, inflammation and gastric cancer achieved 81.4%. PMID- 19810530 TI - [Qualitative and quantitative analysis of Shuanghuanglian fenzhenji by FTIR]. AB - Qualitative and quantitative analysis of Shuanghuanglian fenzhenji was studied in the present paper by FTIR. The fingerprint characteristics of main components in the injections were assigned and can be used as the criterion in the qualitative quality control. Calibration models by PLS algorithm were built to get the quantitative information of baicalin and chlorogenic acid in the injection samples. The R2 of calibration models are both above 0.99 and the average relative deviations between predicted and actual value measured by HPLC are less than 4% for the both properties. The qualitative and quantitative analysis by FTIR is simple and rapid, so it may provide a new on-line approach to the quality control of main constituent in Shuanghuanglian fenzhenji and some other traditional Chinese medicines. PMID- 19810531 TI - [Study of reference and order parameter of Raman spectroscopy research on alkane phase transition]. AB - Raman spectroscopy was used to investigate the order-disorder phase transition of n-eicosane for real-time cooling run. At the phase transition temperature, the integrated intensity of the 1 300 cm(-1) spectra range changes greatly, which requires further consideration of 1 300 cm(-1) as a reference in the research on polymer and biomembrane using Raman spectroscopy. The experiment support was provided for doubt about order parameter based on 1 130 cm(-1) that it did not refer to generally accepted interpretation of an order parameter and it proved that the order definition could just be used as a relative measure method for disorder. PMID- 19810532 TI - [Temperature dependence of additional peaks characteristics of single mode silica fiber stimulated Raman scattering]. AB - Ten meter single mode silica fiber was used to study the temperature characteristics of stimulated Raman scattering (SRS), and additional peaks (double-humped structure) were observed at both sides of pump light and first order Stokes light in the experiment. The peak intensity increased first, and then decreased as the temperature increased from 80 K to 295 K. The first-order Stokes double-humped wave peaks disappeared when the temperature was 295 K. The double-humped peaks phenomenon was caused by simulated four photon mixing (SFPM), according to stimulated four-photon mixing theoretical calculation. At the same time, the phenomenon that the frequency shift of first-order Stokes spectrum line in SRS increased from 706.9 to 712.9 cm(-1) and its half width increased from 1.75 to 2.18 nm was theoretically explained, and the theoretical results are well consistent with experiments. PMID- 19810533 TI - [Discovery of anatase in atmospheric inhalable particles and its significance]. AB - Laser Raman microprobe (LRM) is a reliable technique for phase identification to analyze the molecular composition and microstructure on 1 microm2 area of samples, and therefore, it is well-suited for identifying the mineral phases of single fine particles. In the present paper, we utilized LRM to identify the mineral phases of the single inhalable particles (PM10) from samples in Beijing City and compared the Raman microscopic spectra of samples with the standard spectra of mineral and inorganic material of Renishaw's database. Then we confirmed, for the first time, that the mineral phase of Ti-rich particles in the environmental atmosphere is the anatase TiO2, whose Raman spectrum has a strong O Ti-O band at 638 cm(-1) and two medium O-Ti-O bands at 398 and 517 cm(-1) respectively. Thus it ensures the existence of TiO2 particles in PM10. Anatase is an important photocatalyst which can speed up the heterogeneous reaction between mineral particles, especially the calcium carbonates, when carried by these particles. Furthermore, the crystal structure of anatase, relative humidity of environment and the surface pH value can significantly influence the photocatalysis of anatase in atmosphere. PMID- 19810534 TI - [Evidence of discontinuities in the Raman spectra of aqueous NaCl solution at high pressure]. AB - In-situ Raman spectra measurement for aqueous NaCl solution was conducted at the temperature of 21 degrees C and the pressures of 50-1 100 MPa using a SiC anvil cell. It is shown that the decomposed bands of aqueous NaCl solution shift to lower wavenumber with increasing pressure initially and reaches the minimum at about 300 MPa, and increases at higher pressure up to about 800 MPa, then decreases again with increasing pressure. Similarly, the ratio of band-area and the width at half maximum of the decomposed bands of the solution exhibit discontinuities at about 300 and 800 MPa. This finding demonstrates that the structure of aqueous NaCl solution is discontinuous at high pressure and O-H...Cl bonds change correspondingly, which suggests the existence of rearrangement and the appearance of more complicated configuration in the interior structrure of aqueous NaCl solution. PMID- 19810535 TI - [Raman spectra analysis of Nd : YAG single crystal and its nano-powder]. AB - In the present paper, the authors measured the Raman spectra of YAG/Nd : YAG single crystal, Nd : YAG precursor and the powder sintered at different temperatures. The bands of these Raman spectra were assigned and analyzed. The results show that there is a structure transformation process in the course of sintering Nd : YAG precursor. The powder sintered at 700 degrees C was amorphous and it is of AlO4 tetrahedron structure. With the increase in sintering temperatures, the Raman spectra varied mainly in two respects. One is the decrease in FWHM with the increase in the bands intensity; the other is the bands shift. These should be due to the increase in the order degree of the interface component. Additionally, the difference in the lattice vibration modes between the powders sintered at 800 degrees C and the Nd : YAG single crystal powder was caused by the contribution of the interface component. PMID- 19810536 TI - [Study of the relationship between apoptosis and intracellular pH in single living cells using a three-channel real-time fluorescence imaging method]. AB - Using a Dual-View wavelength splitter, a custom-made filter block and only a single intensified charge-coupled device (ICCD), a three-channel real-time fluorescence imaging method for single living cell research was established based on an ICCD fast fluorescence micro-imaging system. The relevant calibrating method for images was also developed to eliminate the influence introduced by spectral crosstalk. Double-labeled with two fluorescent indicators, Annexin V FITC and SNARF-1, mouse thymocyte was treated with S-nitrosoglutathione (GSNO) and the relationship between their apoptosis and intracellular pH changes was studied in real-time at the single living cell level. The results not only showed the different regularities of intracellular pH changes between apoptosis induced by GSNO and spontaneous apoptosis, but also provided evidence for the application of the three-channel real-time fluorescence imaging method to bio-medical studies. PMID- 19810537 TI - [Measurement of near-UV absorption cross sections of CS2]. AB - The UV-absorption cross sections of CS2 are reported. The absorption spectra were measured by using a diode array spectrometer over the wavelength range of 220-380 nm at room temperature (298 +/- 3)K. The maximum absorption cross section appeared around 315 nm with the value of sigma (315 nm) = 7.184 x 10(-20) cm2 x molecule(-1). The measured data by this study were in good agreement with that reported in the literature, and used to estimate the photolysis of the CS2 in the atmosphere. PMID- 19810538 TI - [Preparation of nanocrystalline goethite (alpha-FeOOH) by gel-network precipitation method and spectral properties]. AB - Iron oxyhydroxides (FeOOH), as an environmental mineral material, can adsorb and coprecipitate the contamination from the medium. The ability of removing contamination is decided by the morphology and structural characteristic and the synthesis methods of the obtained mineral. In the present, the used synthesis methods of iron oxyhydroxides (FeOOH) include ferric iron hydrolyzation and ferrous chemical oxidation. But the products of iron oxyhydroxides prepared by these two chemical methods are easily agglomerated and form bigger particles. Thus, in the present study, a novel gel-network precipitation method was developed to synthesize the nanoparticles of goethite (alpha-FeOOH) as environmental mineral material. During formation of alpha-FeOOH nanoparticles by this method, FeCl3 acted as the reaction material and glutin played a role of the reaction medium, which prevents the presence of agglomeration of precipitate particles. So the obtained alpha-FeOOH nanoparticles had smaller size, no aggregation and basic monodispersity, compared with that prepared by the coprecipitation method. At the same time, we introduced the spectrum analysis measures, and studied the effect of different concentration of glutin and FeCl3 solution on the crystallizability and morphology of products. The structure and morphology of alpha-FeOOH nanocrystallites were determined by means of XRD, FTIR and SEM. The results of the spectrum analysis showed that the particle sizes and shapes and crystallizability of the obtained alpha-FeOOH precipitate products were highly related to the network structure of gelatin. And the goethite particles with better monodispersity, prepared by the optimum concentrations of glutin (12%) and FeCl3 solution (0.6 mol x L(-1)), had a short rod-type shape approximately 110 nm in length with an average diameter of about 35 nm. PMID- 19810539 TI - [Study on the modeling of hyperspectral polarized reflection of clove lamina with chlorophyll content]. AB - In the present study, the authors detected clove laminas in different states with multi-angle hyperspectral polarized reflections and measured their chlorophyll content at the same time. The authors analyzed hyperspectral polarized reflections of clove laminas from various viewing zenith angles, incidence angles, the relative azimuth angles, polarized states and chlorophyll content. The authors calculated quantitatively clove laminas in different states with multi-angle hyperspectral polarized reflection by USB2000, bidirectional polarized reflectance and polarized equipment, and built the regression models of polarized information-chlorophyll content. The result indicated that when the polarized angle was 0 degrees, the model of chlorophyll content with polarization reflectance was built as Y = 4. 506 4e(-0.56 8x) (R2 = 0.895 8); while as the polarized angle was 90 degrees, the model of chlorophyll content with polarization reflectance was built as Y = 145.79X(-1.204 1)(R2 = 0.479 8); when the incidence angle was 50 degrees, the model of chlorophyll content with degree of polarization was built as Y = 7 206.7X(6) -20 160 X(5) + 22 547X(4)-12 788X(5) + 3 822.4X(2) -553.72X + 30.429 (R2 = 0.646 4). The authors found that a significant functional relationship between the polarized information and chlorophyll content exists. The study provides a theoretical base for vegetation remote sensing. PMID- 19810540 TI - [Determination of DNA using Eu-PPA as fluorescence probe by time-resolved fluorescence]. AB - In the Tris-HCl buffer solution, europium ion combined with pipemidic acid (PPA) to form a Eu-PPA complex. The fluorescence intensity was dramatically enhanced when herring sperm DNA was added in the Eu-PPA system. Furthermore, the fluorescence intensity was increased linearly with the concentrations of added DNA within a proper range. Based on this, a simple new method for the determination of herring sperm DNA was developed. Time-resolved fluorescence excitation and emission spectra and the fluorescence lifetime of the system were studied in detail and compared with that from conventional fluorescence method. The experimental results indicated that the time-resolved fluorescence method was superior over the conventional fluorescence method. The reaction conditions, measurement parameters and the adding order of reactants were optimized. The best concentration of both europium ion standard solution and PPA standard solution was 1.00 x 10(-5) mol x L(-1) in the Tris-HCl buffer solution of 0.005 mol x L( 1) with the optimal pH value of 7.2. A detection limit (3 s) of 0.03 mg x L(-1) was obtained with a RSD of 0.3% (4.0 mg x L(-1), n = 11). The linear relationship between deltaI = 89.58c (mg x L(-1)) +0.920 5 and the regression factor of 0.999 6 was obtained in the range of 0.1-6 mg x L(-1). This method was applied to the determination of DNA in synthetic samples. The results and recoveries of spiked standard solution in the sample were satisfied. PMID- 19810541 TI - [Spectroscopic investigation on the interaction of protocatechuic acid and veratric acid with biomacromolecules]. AB - Protocatechuic acid (P) and veratric acid (V) are phenolic acidic compounds and have a wide biological and pharmaceutical activities, and their interaction with biomacromolecule has been a hot topic. The interaction mechanism of P and V with fsDNA was investigated by fluorescence and UV absorption spectroscopic methods. The UV results showed that P and V have three strong absorption bands at 190-230 nm (K band), 230-270 nm (B band) and 270-310 nm (R band) respectively. When the excitation wavelength was 280 nm, the fluorescence emission bands of P and V were at 338 and 334 nm, respectively, while the fluorescence emission band of DNA was very weak and had little influence on those of the P and V. The fluorescence intensities of the P and V were strongly quenched after interacting with fsDNA, and their Stern-Volmer quenching rate constants were 1.03 x 10(12) and 0.61 x 10(12) L x mol(-1) x s(-1), respectively. It was illustrated that the fluorescence quenching was mainly static and the complex was formed between the drug and fsDNA. When the concentration of DNA was high, their Stern-Volmer curves were not linear, and it was indicated that the quenching mechanism was complex and may contain dynamic quenching process. Their binding constants were calculated based on the static fluorescence quenching, with K(fsDNA/P) = 6.22 x 10(6) L x mol(-1) and K(fsDNA/v) = 1.57 x 10(4) L x mol(-1). The investigation showed that the molecular ratio of V-fsDNA was 1 : 1, while that of P-fsDNA was 1: 2. It was demonstrated that the protocatechuic acid can bind with two bases of fsDNA, which was related to the two hydroxyl groups on the drug molecule. The results showed that the structure of P and V greatly influenced their binding mode with DNA molecules. PMID- 19810542 TI - [SPAD prediction of leave based on reflection spectroscopy]. AB - Handheld SPAD meter is often used to measure chlorophyll content of plant and nitrogen level for some species. For plant production automation, however, it loses its popularity due to its point-by-point checking. The authors need to monitor the growing conditions of plant remotely, instantly and nondestructively. In the test, we examined optical fiber reflection spectroscopy used to measure chlorophyll content of some plant leaves, or for their SPAD prediction. The authors picked 120 leaves randomly from our campus ground or trees, among which 70 samples were chosen as calibration set and others as verification set. Each sample was water-cleaned and air-dried. To locate each measuring point precisely when using SPAD meter and spectrometer, the authors drew a circle with a diameter of 10 mm on each leave to be measured. By comparing the spectral curves of various leaves, the authors found that the spectral band between 650-750 nm was significant for SPAD modeling since this range of spectral data of leaves with the same SPAD reading was close to each other. It was showed that leave color was an unnecessary factor for SPAD prediction by reflection spectroscopy. Besides, the authors discovered that LED's narrow spectral range used by SPAD meter should be concerned because optical fiber spectrometer has much more wide spectral range. Based on this awareness, the authors designed an adjustment factor of light to linearly rebuild spectrometer's reflective intensity so that it reached zero outside the band 650-750 nm. Moreover, leave thickness was another influential factor for SPAD prediction since the light of SPAD meter goes through the leave while the reflective spectrometer does not. First, an equation for SPAD prediction was built with uncertain parameters. Then, a standard genetic algorithm was designed with Visual Basic 6.0 for parameter optimization. As a result, the optimal reflection band was narrowed within 683.24-733.91 nm. The result showed that leave thickness strongly affects the precision of SPAD prediction. Through the modification of leave thickness, the regression coefficient (R2) of calibration set and verification set reached 0.865 8 and 0.916 1 respectively. The test showed that optical fiber reflection spectroscopy is useful for SPAD prediction and can be used to develop remote SPAD sensor. PMID- 19810543 TI - [Advance in nondestructive detection of fruit internal quality based on hyperspectral imaging]. AB - The present paper reviews the development in the field of hyperspectral imaging technology for nondestructive detection of fruit internal quality in recent years up to the year 2007. With the increasing maturity of hyperspectral imaging technology, decline of cost for its hardware and software, and improvement in hyperspectral image data processing algorithms, hyperspectral imaging technology for fruit quality nondestructive detection has become a hot research topic. In order to track the latest research developments at home and abroad, the fruit internal quality (maturity, firmness, soluble solid content, water content) detection with hyperspectral imaging was reviewed, which would provide reference for Chinese researchers. PMID- 19810544 TI - [Review of digital ground object spectral library]. AB - A higher spectral resolution is the main direction of developing remote sensing technology, and it is quite important to set up the digital ground object reflectance spectral database library, one of fundamental research fields in remote sensing application. Remote sensing application has been increasingly relying on ground object spectral characteristics, and quantitative analysis has been developed to a new stage. The present article summarized and systematically introduced the research status quo and development trend of digital ground object reflectance spectral libraries at home and in the world in recent years. Introducing the spectral libraries has been established, including desertification spectral database library, plants spectral database library, geological spectral database library, soil spectral database library, minerals spectral database library, cloud spectral database library, snow spectral database library, the atmosphere spectral database library, rocks spectral database library, water spectral database library, meteorites spectral database library, moon rock spectral database library, and man-made materials spectral database library, mixture spectral database library, volatile compounds spectral database library, and liquids spectral database library. In the process of establishing spectral database libraries, there have been some problems, such as the lack of uniform national spectral database standard and uniform standards for the ground object features as well as the comparability between different databases. In addition, data sharing mechanism can not be carried out, etc. This article also put forward some suggestions on those problems. PMID- 19810545 TI - [Fabrication and photocatalytic activity of Pt-inserted titania nanotubes]. AB - Titania nanotubes (TNTs) were synthesized by hydrothermal treatment of rutile phase TiO2 nanoparticals in NaOH solution at 110 degrees C for 24 hours. After drying in aceton for 36 h, the TNTs were under vacuum drying for 24 h at room temperature. The Pt-inserted titania nanotubes (Pt/TNTs) were obtained by filling H2 PtCl6 ethanol solution into the TNTs after vacuum drying. The characterizations of the as-synthesized samples were confirmed by TEM, XRD, and UV-Vis. The photocatalytic activity of the Pt/TNTs was investigated by photo induced decomposition of methyl orange(MO)under the main 365 nm UV-light. In order to comparison, the photocatalytic activity of both the rutile-phase TiO2 nanoparticles and pure TNTs were also investigated at the same time under the same experimental conditions. The TEM images show that the TNTs are hollow, a few hundred nanometers long, and the inner/outer diameter is about 6/10 nm. The crystal structure of TNTs is H2Ti2O5 x H2O with a little Na. Both the shape and the crystalline of the TNTs are not changed after the modification. The oval or round Pt0 nanoparticals, about 3 nm in diameter, are found only in the nanotubes. Pt/TNTs exhibit enhanced absorption at the visible range in the UV-Vis spectra and its start absorption band edge(lambda0 approximately 457 nm)is obviously redshifted compared to the rutile-phase TiO2 nanoparticals and pure TNTs. The Pt nanoparticles are found to significantly enhance the photocatalytic activity of TNTs. Pt/TNTs are demonstrated to be highly efficient for the UV-light induced photocatalytic decomposition of MO compared to both the rutile-phase TiO2 nanoparticals and pure TNTs. After irradiation for 60 min, the photocatalysis decomposition rate of MO in rutile-phase TiO2 nanoparticals, TNTs and Pt/TNTs are 46.8%, 57.2% and 84.6% respectively. PMID- 19810546 TI - [Detecting land use change using PCA-enhancement and multi-source classifier from SPOT images]. AB - Concomitant with the rapid global urbanization process, land use change detection has been the focus and "hot spot" of global change research all the time. In the present study, the rigorous orthorectification was first applied to the SPOT-5 data to guarantee precise geometric correction and image registration. Afterwards, a methodology integrating PCA-enhancement and multi-source classifier was adopted to detect the land use changes in urban area. The results show that the first three PCs derived from multi-temporal-PCA contain most of the spectral information among which unchanged land use is highlighted in PC1 and PC2, and changed land use is mainly enhanced in PC3. The following multi-source classifier integrating unsupervised classifier (ISODATA) and supervised classifier (Maximum Likelihood) accurately extracts all the information. The findings from accuracy assessment demonstrate that the overall accuracy for the proposed method reaches 92.58, KAPPA coefficient is 0.92, and proving figures are also produced for the user's and producer's accuracies. It was further found that the proposed method yielded better accuracy than that of traditional post-classification comparison approach. PMID- 19810547 TI - [Time resolved UV-Vis absorption spectra of quercetin reacting with various concentrations of sodium hydroxide]. AB - A real time investigation of chemical reaction process of quercetin with various concentrations of sodium hydroxide was performed by using an intensified spectroscopic detector ICCD. The time resolved UV-Vis absorption spectra of 5 x 10(-5) mol x L(-1) quercetin respectively reacting with sodium hydroxide at concentrations of 2, 0.2, 0.1, 0.04 and 0.02 mol x L(-1) were acquired. A total of 200 spectra with the same exposure time of 0.1 ms for each spectrum but different time interval between two consecutive spectra were recorded for each reaction. The first 50 spectra have the time interval of 20 ms, the next 50 have 1 s, and the last 100 have 2 s. Results indicate that quercetin reacted with sodium hydroxide easily and there was an intermediate product formed during the reaction, with different concentrations of reactants, the changes of absorption bands were the same, but the moments at which the changes happened were different and the total reaction time was various from 1 s to 100 s. Spectra recorded showed the disappearing process of the typical bands centered at 254 and 374 nm of pure quercetin, the growing and disappearing processes of a new band centered at 427 nm of the intermediate product, and the growing process of the new band centered at 314 nm of the final product obviously. No other transient spectroscopic data are currently available on the reaction of quercrtin with sodium hydroxide, the results obtained in the present work provide useful experimental data for the study of the microscopic process of the reaction. PMID- 19810548 TI - [Application of hyperspectral remote sensing in research on ecological boundary in north farming-pasturing transition in China]. AB - Studies of ecological boundaries are important and have become a rapidly evolving part of contemporary ecology. The ecotones are dynamic and play several functional roles in ecosystem dynamics, and the changes in their locations can be used as an indicator of environment changes, and for these reasons, ecotones have recently become a focus of investigation of landscape ecology and global climate change. As the interest in ecotone increases, there is an increased need for formal techniques to detect it. Hence, to better study and understand the functional roles and dynamics of ecotones in ecosystem, we need quantitative methods to characterize them. In the semi-arid region of northern China, there exists a farming-pasturing transition resulting from grassland reclamation and deforestation. With the fragmentation of grassland landscape, the structure and function of the grassland ecosystem are changing. Given this perspective; new image processing approaches are needed to focus on transition themselves. Hyperspectral remote sensing data, compared with wide-band remote sensing data, has the advantage of high spectral resolution. Hyperspectral remote sensing can be used to visualize transitional zones and to detect ecotone based on surface properties (e. g. vegetation, soil type, and soil moisture etc). In this paper, the methods of hyperspectral remote sensing information processing, spectral analysis and its application in detecting the vegetation classifications, vegetation growth state, estimating the canopy biochemical characteristics, soil moisture, soil organic matter etc are reviewed in detail. Finally the paper involves further application of hyperspectral remote sensing information in research on local climate in ecological boundary in north farming-pasturing transition in China. PMID- 19810549 TI - [A new immuno-nanosliver resonance scattering spectral probe for assay of IgG]. AB - Using tri-sodium citrate as reducer, stable silver nanoparticles the size of about 20 nm were prepared by the microwave high pressure procedure with simplicity and rapidity. At pH 9.0, sliver nanoparticle was used to label goat anti-human IgG (GIgG) to obtain an immuno-nanosilver resonance scattering spectral probe (AgGIgG) for IgG. In pH 6.0 buffer solution and in the presence of polythylene glycol (PEG) and KCl, the immune reaction of IgG with AgGIgG took place, the silver nanoparticles released from AgGIgG produced aggregations, and the resonance scattering intensity at 485 nm (I(485 nm)) was enhanced greatly. The influence factors such as pH value, buffer solution volume, concentration of AgGIgG, KCl, PEG-4000, PEG-6000, PEG-10000 and PEG-20000, incubation temperature and time were considered, respectively. Under the conditions of 0.40 mL of pH 6.0 phosphate buffer solution, 1.20 mL of 9.8 microg x mL(-1) AgGIgG, 0.20 mL of 20% PEG-6000, 0.50 mL of 10% KCl, and ultrasonic irradiation for 25 min at room temperature, the increased intensity deltaI(485 nm), was proportional to the IgG concentration (c(IgG)) from 0.004 to 0.48 microg x mL(-1), with a detection limit of 2.4 ng x mL(-1). The regress equation was deltaI485 nm = 76.8c(IgG) + 4.7. The effect of foreign substances such as 20 microg x mL(-1) Ni2+, Fe2+, Pb2+ and BSA,60 microg x mL(-1) Cu2+, Ca2+ and HSA,60 microg x mL(-1) Mg2+ and Mn2+, 320 microg x mL(-1) Zn2+, glucose and urea on the deltaI(485 nm) was examined, respectively. Results showed that there was no interference. This assay showed high sensitivity and good selectivity for quantitative determination of IgG in human serum with satisfactory results. The analytical results were in agreement with that of the reference results. PMID- 19810550 TI - [Determination of the daily changes curve of nitrogen oxides in the atmosphere by digital imaging colorimetry method]. AB - From the digital images of the red complex which resulted in the interaction of nitrite with N-(1-naphthyl) ethylenediamine dihydrochloride and P-Aminobenzene sulfonic acid, it could be seen that the solution colors obviously increased with increasing the concentration of nitrite ion. The JPEG format of the digital images was transformed into gray-scale format by origin 7.0 software, and the gray values were measured with scion image software. It could be seen that the gray values of the digital image obviously increased with increasing the concentration of nitrite ion, too. Thus a novel digital imaging colorimetric (DIC) method to determine nitrogen oxides (NO(x)) contents in air was developed. Based on the red, green and blue (RGB) tricolor theory, the principle of the digital imaging colorimetric method and the influential factors on digital imaging were discussed. The present method was successfully applied to the determination of the daily changes curve of nitrogen oxides in the atmosphere and NO2- in synthetic samples with the recovery of 97.3%-104.0%, and the relative standard deviation (RSD) was less than 5.0%. The results of the determination were consistent with those obtained by spectrophotometric method. PMID- 19810551 TI - [Differentially expressed protein study on unstable angina blood-stasis syndrome by fluorescence labelling method]. AB - Fluorescence labelling method was used seeking differentially expressed plasma proteins of unstable angina blood-stasis syndrome. Three fluorescence labelling dyes (Cy2, Cy3 and Cy5) were used on two classes of species (plasma of 12 patients with unstable angina blood-stasis syndrome and 12 healthy volunteers). Two dimensional gel electrophoresis was carried out, and three different fluorescence scans were performed. By analyzing the graphs and mass spectrometry, the differentially expressed plasma proteins of unstable angina were obtained. Blood-stasis syndrome. Results show that fibrinogen beta chain, fibrinogen gamma chain, alpha1-antitrypsin, haptoglobin beta chain and haptoglobin alpha2 chain were significantly highly expressed in the plasma of unstable angina blood-stasis syndrome patient, while ApoA-IV, ApoA-I and transthyretin were decreased in the plasma of unstable angina blood-stasis syndrome patient. These identified proteins could be divided into two categories according to their functions: (1) Acute phase reactive protein; (2) Apolipoprotein. In conclusion, fibrinogen beta chain, fibrinogen gamma chain, alpha1-antitrypsin, transthyretinc, haptoglobin beta chain, ApoA-IV, ApoA-I and haptoglobin alpha2 chain are differentially expressed in the patients with unstable angina blood-stasis syndrome and control group, some differentially expressed proteins were correlated with inflammatory reaction or lipid metabolic disorder, and these proteins could provide clues to the study and discovery of new protein targets for antianginal drugs. The fluorescence labelling method is suitable for discovery of differentially expressed proteins. PMID- 19810552 TI - [Research on the autofluorescence spectroscopy of heart tissues]. AB - The present study investigated the three-dimensional spectra and emission spectra of the autofluorescence of rabbit hearts. The results suggested that the three dimensional spectra of the iced atria and ventricle were observed more evidently different from that of the fresh tissue compared to the main artery, which indicated that the amount of flavins and NADHs changed. Also, the atria, ventricle and main artery have different specific excitation spectra at the wavelength of 340 nm. The main fluorescence peaks were of NADH (at about 460 nm), collagen and elastin (at about 290-400 nm). The Gauss spectra of atria and ventricle were different in the peak value, relative intensity and half width. So the ratios of fluorescence intensities of peaks may be used to distinguish different heart tissues. Furthermore, a phenomenon was firstly uncovered that the autofluorescence intensity of NADH in ventricle decays with the time of death and it could be a useful method for the estimation of postmortem interval. PMID- 19810553 TI - [Enteromorpha prolifera underwater spectral research based on simulation of radiation transmission]. AB - The accumulation of Enteromorpha prolifera in huge amount in the Yellow sea in June, 2008 draws the attention from all over the world. It is an urgent requirement to monitor the wide range of Enteromorpha prolifera distribution by remote sensing. As to the Enteromorpha prolifera floating on the sea surface, effective monitoring by optical remote sensing has been basically achieved. As far as the underwater suspended Enteromorpha prolifera is concerned, the present paper carried out the radiative transfer simulation research on the above water spectral response, its variation with the suspending depth, the water turbidity and environmental conditions. It was found that with the increase in Enteromorpha prolifera suspending depth and water turbidity as well as the decrease in the thickness of Enteromorpha prolifera, the Enteromorpha prolifera information contained in the surface spectra would decrease. The influence of environmental factors such as water-gas interface roughness, cloud cover extent and sun zenith angle on the underwater suspended Enteromorpha prolifera spectra can be ignored. The maximum Enteromorpha prolifera depth that can cause surface spectrum changes is about 30 m in clean water and about 1 m in turbidity water. PMID- 19810554 TI - [Degradation of organic contaminant in landfill leachate by photo-Fenton process]. AB - In the present work, the treatment of landfill leachate was studied by photo Fenton process. All photocatalytic experiments were carried out under similar conditions on July or August sunny days between 12 a. m. and 14 p. m. in Chongqing based on the change in solar light intensity with time. The effect of operating conditions such as Fe/H2O2, pH value and Fenton's reagent dosage on color removal and UV254 was investigated. The UV-Vis absorption spectra of 1.00 X 10(3) mg x L(-1) landfill leachate were considered before and after photo-Fenton experiment. The experiment result indicated that photo-Fenton process can effectively remove color. The oxidation of organic materials in the leachate was pH dependent and the optimal pH was 2.5. The favorable Fe2+ to H2O2 molar ratio was 1/95. The optimal conditions in this research were Fe2+ concentration of 5.00 mmol x L(-1) and H2O2 concentration of 5.70 x 10(2) mmol x L(-1). Meanwhile, the analysis of reaction kinesics was carried out in the research. Second-order kinetic was observed for the degradation of landfill leachate, and the rate equation of photo-Fenton process was -dc/dt = -4.34[c]1.92. The result of UV-Vis spectra of landfill leachate showed that photo-Fenton process is an effective method for removal of organic compounds. PMID- 19810555 TI - [Spectroscopic investigation of the poly[(silylene)acetylene silanes]]. AB - In the present paper, a series of novel conjugated polymers, poly[(silylene)acetylene silanes] with different structure, were prepared, and the luminescence performance of the poly[(silylene)acetylene silanes] with different substituents were investigated by the fluorescence and UV absorption spectroscopic approaches. The effect of different substituents and the number of acetylene in the main chain on the luminescence was discussed in depth. The result showed that these polymers have moderate absorbance in the range from 219 to 260 nm. The red shift was showed for the maximum absorbance wavelength following the increase in the number of the acetylene and the degree of the conjugation in the main chain. There was insignificant difference in the maximum absorbance wavelength of the polymers with dimethy and diphenyl. In short, the influence of the substituents is insignificant. However, the remarkable effect was induced by the conjugated structure in the main chain. The conjugated polymers with different structure in the main chain have moderate fluorescence and emission quantum yields. The authors studied the influence of the polymers' structure on the luminescence performance. As a result, the influence of the substituents is insignificant for emission spectra. The influence of the conjugated groups in the main chain of the polymers is remarkable. The maximum emission wavelength of the polymers showed an evident shift to red range with the enhancement of the conjugated extent. These polymers had good thermal properties for the special structures. So they have potential applications for emission materials with good thermal stability. PMID- 19810556 TI - [Analysis of the extraction of red pellicle of walnut (Juglans regia L.) by ultraviolet-visible spectra and HPLC-ESI-MS(n)]. AB - The extraction of the red pellicle of walnut (Juglans regia L.) was analyzed by UV-visible spectra and HPLC-ESI-MS(n) (high-performance-liquid-chromatography coupled with electrospray ionization mass spectrometry). The extraction in ethanol-HCl showed two absorption peaks at 560 and 591 nm respectively in the UV Vis spectrum; after purified by lead acetate and thin-layer-chromatography, the extraction in ethanol-HCl showed 4 absorption peaks at 340, 370, 552 and 585 nm respectively. These results testified that the anthocyanin was in the extraction. Six molecular ion peaks (m/z) occurred on MS: 301, 481, 633, 783, 785 and 950, which was identified as ellagic acid, Hexahydroxydiphenoyl(HHDP)-glucose, Galloyl HHDP-glucose, Di-HHDP-glucose, Di-Galloyl-HHDP-glucose, and HHDP-Valoneoyl glucose respectively. PMID- 19810557 TI - [LED illuminant-based detection of trace NO2 gases]. AB - Light-emitting diode was used as illuminant in the present paper. The light emitting diode was chosen according to the high absorption in the range of absorption spectrum of NO2. The pollutant concentration can be calculated by measuring the absorption spectrum, and fitting the experimental absorption spectrum and normal absorption spectrum by means of least square fitting method. The method of measuring nitrogen dioxide concentration making use of absorption spectra was studied, the basic principle was analyzed, the process of data analysis was studied including the fitting of experimental spectrum and standard spectrum, wiping off the slow change, using least-square fitting in reverse calculating of concentration etc. The detecting precision was improved by the method of absorption spectrum. In this experiment the light emitting diode was used as illuminant, which made the detecting become not only convenient but also fast, and online real time supervising can be realized. Taking advantage of the characteristic that NO2 has strong absorption in the range of 300-500nm, the NO2 concentration of sample was measured. PMID- 19810558 TI - [Determination of as in industrial wastewater by laser-induced breakdown spectroscopy]. AB - The wastewater from industrial smelting process contains heavy metals such as arsenic (As) that produce serious environmental pollution and cause actual harm to the health of people. It is necessary to control the pollution at the source and achieve a real-time and online monitoring. The laser-induced breakdown spectroscopy (LIBS) is a new elemental analysis technique, and has the advantage of rapid detection. An LIBS setup has been established. The Nd : YAG laser beam is focused onto the sample, then the plasmas are produced. The emission spectra of plasmas are dispersed by an Echelle spectrograph and detected by an intensified charge-coupled device (ICCD). Experiments have been carried out on the industrial wastewater collected from the scene. The spectral lines of As element were obtained. The calibration curve of the line intensities versus the concentrations of the As element was acquired by the experiment. The calibration curve can be used for the quantitative analysis of arsenic element with an unknown concentration in the industrial wastewater. The results showed that the LIBS technique can be applied in the rapid detection of As element in industrial wastewater, and has wide range of applications. PMID- 19810559 TI - [Quantitative analysis of the concentration of Br-doping in micro-shell coating with XRF]. AB - In inertial confinement fusion (ICF) physics experiment, the micro-shell that contains Br-doped CH coating must be characterized for doping Br concentration level. X-ray fluorescence (XRF), with its unique capability to quantitatively determine concentrations of most elements simultaneously and non-destructively, is generally the method of choice for total dopant (Z > 11) concentration. In the present paper, a method to determine the dopant concentration in ICF micro-shell coating with X-ray fluorescence spectrometry is described, and the calibration model is founded by the calculation of fluorescence intensity of film and micro shell sample. Based on the calibration model, the fluorescence intensity vs concentration of Br-doped CH coating of micro-shell was obtained. The experiment result shows that X-ray fluorescence spectrometry is a nondestructive and accurate method of measurement of coating dopant in the inertial confinement fusion micro-shell sample, and the measuring error is about 5% for Br doped CH coating of micro-shells with 10 micron thickness coating. PMID- 19810560 TI - [Comparison of four kinds of measurement techniques for wood microfibril angle]. AB - X-ray diffraction technique, pit aperture observing technique, polarized light microscope technique and near infrared spectrum technique were used separately to test the wood microfibril angle of Chinese fir, and the results were as follows: For the identical tree of Chinese fir, the average microfibril angle obtained by the polarized light microscope technique was the biggest, followed by those obtained by X-ray diffraction technique, and then by polarized light microscope technique, but the difference in the average microfibril angle was not significant. For the different annual rings, the microfibril angle obtained by X ray diffraction technique became the biggest after the 20th annual ring, the fluctuation of the microfibril angle got by the pit aperture observing technique were big, but the fluctuation got by the polarized light microscope technique was just the reverse. The measurement value curves got by the three techniques were fitted well, and the difference in the average microfibril angle was not significant between different annual rings. For the microfibril angle of the same annual ring, the difference between the maximum and the minimum value tested by the polarized light microscope technique was less than 4 degrees, but the difference tested by the pit aperture observing technique achieved 21.53 degrees, and the standard deviation achieved 4.75. The near infrared spectrum and the Xray diffraction techniques were all called nondestructive testing techniques. The model set up by the two techniques was very good because of its high prediction and repeatability, and the model was also good for online analysis. The R2 of calibration model and tested model achieved 0.81 and 0.75 respectively,and the standard error of calibration and prediction were 1.79 and 2.02 respectively. In addition, the near infrared spectrum technique could be also used with the other three techniques to predict the wood microfibril angle, showing the superiority of the near infrared spectrum technique. Meanwhile, the advantages and shortcomings were analyzed for the four kinds of measurement techniques. PMID- 19810561 TI - [Determination of trace elements in electrical absorption prospecting polyform sample by inductively coupled plasma mass spectrometry]. AB - An ICP-MS method was established for the determination of sixteen trace elements, V, Cr, Mn, Co, Ni, Cu, Zn, Ga, Nb, Mo, Ag, Cd, Au, Tl, Pb and Bi in electrical absorption prospecting polyform. Three methods for polyform samples (ashing method, extraction by HNO3 + H2O2 and digestion with aqua regia) were compared and the results showed that the second method is the best one. The best operational paramenters of X series ICP-MS were confirmed, the inner standard 103Rh and 185Re were selected for the determination of elements, and analysis of isotopes interference correction equations was established. Satisfactory linearity of working curves of the sixteen trace elements was obtained, giving all their correlation coefficients over 0.999 8. The determination limit of the analytes was in the range of 0.001-2.2 microg x g(-1). The precision was 1.39% 4.84%, and the recoveries were between 94.86% and 105.2%. The method is sensitive, quick and simple and has been applied to the analysis of a great number of polyform samples. PMID- 19810562 TI - [The zonal characterization of elemental concentrations in brown rice of core collection for rice landrace in Yunnan Province by ICP-AES]. AB - In the present paper, the contents of 8 elements in brown rice of 789 accessions core collection for rice landrace from 16 prefectures of five rice regions in Yunnan province were determined by ICP-AES technique. The method proves to be simple, rapid, highly sensitive and accurate, and can be used to determine many elements at the same time. Its recovery ratio obtained by standard addition method ranged from 97.1% to 110.2%, and its RSD was from 0.7% to 4.4%. The analytical results showed that the elemental concentrations (mg x kg(-1) ) in brown rice are in turn of P(3 834.83 +/- 486.49) > K(2 567.72 +/- 336.74) > Mg (2 567.72 +/- 336.74) > Ca (153.67 +/- 55.90) > Zn( 33.35 +/- 13.65) > Fe(32.08 +/- 25.51) > Cu (14.22 +/- 11.85) > Mn(13.58 +/- 3.22). The highest P content is in brown rice from the northwest Yunnan with the rich nonferrous metals and most abundance of biodiversity in the world, the highest Ca, Mg, Fe and Zn concentrations are in brown rice from the middle Yunnan with early cambrian fauna and phosphorite enrichment, and the highest Cu and Mn contents are in brown rice from the southwest Yunnan with the prominent crop diversity. The distributing zones with the highest P and K, middle Ca, Mg and Mn and lowest Fe and Zn in Yunnan are the enrichment zone of minal resources and largest biodiversity. As far as we know, this is the first report that the zonal characterstics of mineral elemental concentractions in brown rice are associated with a lot of factors, such as biodiversity center, enrichment zone of minal resources, origin of life, mountain ranges and rivers and so on, and further it was deduced that the asymmetry of distribution for mineral elements and its reciprocity of mountain ranges and rivers are the key of origin of life. The above results provided reliable data and theory bases for the malnourished Fe and Zn and Ca for 4 billion people in the world, origin of life, the genetic breeding and production of functional rice. PMID- 19810563 TI - [Content of rare earth elements in wild Hypericum japonicum Thunb]. AB - Rare earth elements are important nutritional elements for human health, and today more and more attention has been paid to the effective components in Chinese traditional medicine, especially to rare earth elements. Fifteen rare earth elements in wild hypericum japonicum Thunb were analyzed by the methods of ICP-MS. The results showed that the concentrations of La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Yb, Tm, Lu and Y ranged from 6 ng x g(-1) x DW to 14 522 ng x g( 1) x DW, and among them the concentrations of La, Ce and Nd were higher than 2 000 ng x g(-1) x DW. Compared with the concentration of rare earth elements in rice, corn, wheat and barley, the total concentration of rare earth elements in hypericum japonicum Thunb was much higher, which could be the mechanism of curative effect of hypericum japonicum Thunb on liverish diseases. The character of elements and the content of rare earth elements in soil should be responsible for the difference, but the distributive mechanism of rare earth elements in hypericum japonicum Thunb should be further studied. PMID- 19810564 TI - [Study on the adsorption behavior of nickel on potassium tetratitanate whisker by flame atomic absorption spectrometry]. AB - The adsorption behavior of Ni (II) on potassium tetratitanate whisker was studied systemically by FAAS, seeing about the reasons for absorption, disentanglement and the effect of interfering ions on the recovery. The preconcentration method was simple, quick and had good selectivity. The adsorption rate of Ni(II) by potassium tetratitanate whisker was 100% at pH 5.0 and Ni(II) could be eluted from potassium tetratitanate whisker with hydrochloric acid (C: 0.5 mol x L(-1)), shake time exceeding 5 min, stick time exceeding 1.5 h, and the disentanglement ratio exceeding 90%. The relative standard deviation (RSD) was 2.6%. The results obtained indicate that the potassium tetratitanate whisker has good regenerate capability. PMID- 19810565 TI - [RVM supervised feature extraction and Seyfert spectra classification]. AB - With recent technological advances in wide field survey astronomy and implementation of several large-scale astronomical survey proposals (e. g. SDSS, 2dF and LAMOST), celestial spectra are becoming very abundant and rich. Therefore, research on automated classification methods based on celestial spectra has been attracting more and more attention in recent years. Feature extraction is a fundamental problem in automated spectral classification, which not only influences the difficulty and complexity of the problem, but also determines the performance of the designed classifying system. The available methods of feature extraction for spectra classification are usually unsupervised, e. g. principal components analysis (PCA), wavelet transform (WT), artificial neural networks (ANN) and Rough Set theory. These methods extract features not by their capability to classify spectra, but by some kind of power to approximate the original celestial spectra. Therefore, the extracted features by these methods usually are not the best ones for classification. In the present work, the authors pointed out the necessary to investigate supervised feature extraction by analyzing the characteristics of the spectra classification research in available literature and the limitations of unsupervised feature extracting methods. And the authors also studied supervised feature extracting based on relevance vector machine (RVM) and its application in Seyfert spectra classification. RVM is a recently introduced method based on Bayesian methodology, automatic relevance determination (ARD), regularization technique and hierarchical priors structure. By this method, the authors can easily fuse the information in training data, the authors' prior knowledge and belief in the problem, etc. And RVM could effectively extract the features and reduce the data based on classifying capability. Extensive experiments show its superior performance in dimensional reduction and feature extraction for Seyfert classification. PMID- 19810566 TI - [Comparison of dispersion parts of conventional spectrometers]. AB - Innovation of conventional spectrometers is of actual technical and economical value. It is also an important way to accelerate the development of spectroscopic instruments. When improving a conventional spectrometer, its dispersion part is pivotal, because it is decisive to the basic performance of the spectrometer. In the present paper, the typical dispersion parts of conventional spectrometers are compared to feature them and find the evolution force among them. The basic characters of the dispersion parts, including spectral range, dispersion power, resolution and throughput, are compared separately and comprehensively by reviewing their decisive factor, formula and typical data. The results not only conclude the feature and the complementariness of the dispersion parts, but also indicate that the trade-off between resolution and throughput is ubiquitous in traditional spectrometers. Further reviewing from this point, the evolution history of traditional spectrometers shows that the conflict between resolution and throughput is an important evolution force. This is a new way to understand the evolution of traditional spectrometers. Moreover, dealing with the trade-off between resolution and throughput correctly will help to analyze and settle the core problem of spectrometers. PMID- 19810567 TI - [A micro-silicon multi-slit spectrophotometer based on MEMS technology]. AB - A new mini-spectrophotometer was developed by adopting micro-silicon slit and pixel segmentation technology, and this spectrophotometer used photoelectron diode array as the detector by the back-dividing-light way. At first, the effect of the spectral bandwidth on the tested absorbance linear correlation was analyzed. A theory for the design of spectrophotometer's slit was brought forward after discussing the relationships between spectrophotometer spectrum band width and pre-and post-slits width. Then, the integrative micro-silicon-slit, which features small volume, high precision, and thin thickness, was manufactured based on the MEMS technology. Finally, a test was carried on linear absorbance solution by this spectrophotometer. The final result showed that the correlation coefficients were larger than 0.999, which means that the new mini spectrophotometer with micro-silicon slit pixel segmentation has an obvious linear correlation. PMID- 19810568 TI - [The analysis of the sampling modulation transfer function and the influence on the Gaussian spectra]. AB - As the parameters of the photoelectric detector have important effects on the performance of the dispersive spectrometers, it is necessary to detail the discrete sampling process of the photoelectric detector array. In the present paper, the sampling model was setup, and the effects on the sampling results caused by the spatial frequency of the cosine signal, the width of the sampling pixel, and the initial phase of the sampling pixel position to the crest of the input cosine signal were discussed thoroughly in the frequency domain. By introducing the integral function, a general expression of the sampling modulation transfer function was given, and the concept and expression of the average sampling modulation transfer function was proposed. Since that expression eliminates the effect of initial phase, it is much more convenient to the practical applications. For the typical Gaussian spectrum produced by the dispersive spectrometer, the Fourier transform result of that spectrum was multiplied by the average sampling modulation transfer function to produce a functional expression of the modulation transfer function of the whole system. The average aliasing error of the sampling process was expressed as a function of spatial frequencies; the relationship between the peak value of the average aliasing error and the width of the Gaussian spectrum was discussed; and the critical value of the spectrum width to restore this spectrum precisely was proposed. That critical value is significant for providing guidance to the design and fabrication of dispersive spectrometers. PMID- 19810569 TI - [Design and simulation of the NIR micro-spectrometer optical system]. AB - Based on the working principle of analyzing spectrometer and the theory of optical design, to make the system miniature, and with a certain spectral range and resolution requirements as specific design objectives, a cross-C-T micro structure based on a plane diffraction grating spectrophotometer of the near infrared spectrometer was put forward; ZEMAX was used for design and optimizing and simulating analysis of micro-near-infrared spectrometer in the optical system. The result showed that the micro-optical system's spectral range was 900 1 700 nm, resolution was < 10 nm, spectrum broadening was 12.74 mm, F number was 8.128 388, and the system volume was 51.26 mmX41.81 mmX22 mm, and these all satisfy the requirements. It solved the micromation of the optical system of the spectrometer, and therefore, contributed to the micromation of the spectrometer itself. PMID- 19810570 TI - Vindication is not the business of science. PMID- 19810571 TI - Yogic exercises and health--a psycho-neuro immunological approach. AB - Relaxation potential of yogic exercises seems to play a vital role in establishing psycho-physical health in reversing the psycho-immunology of emotions under stress based on breath and body awareness. However, mechanism of yogic exercises for restoring health and fitness components operating through psycho-neuro-immunological pathways is unknown. Therefore, a hybrid model of human information processing-psycho-neuroendocrine (HIP-PNE) network has been proposed to reveal the importance of yogic information processing. This study focuses on two major pathways of information processing involving cortical and hypothalamo-pituitary-adrenal axis (HPA) interactions with a deep reach molecular action on cellular, neuro-humoral and immune system in reversing stress mediated diseases. Further, the proposed HIP-PNE model has ample of experimental potential for objective evaluation of yogic view of health and fitness. PMID- 19810572 TI - Ulcer healing properties of ethanolic extract of Eugenia jambolana seed in diabetic rats: study on gastric mucosal defensive factors. AB - Diabetes has been reported to cause an increase in offensive and decrease in defensive gastric mucosal factors, the imbalance of which can cause ulceration and delay the ulcer healing. Eugenia jambolana has been documented to have both antidiabetic and antiulcer activities. The present study evaluates the effects of ethanolic extract of E. jambolana on gastric ulcer healing and on rat gastric mucosal defensive factors in gastric ulcer with co-occurring diabetes. E. jambolana extract was administered orally in the dose of 200 mg/kg once daily for 10 days. E. jambolana extract increased mucin secretion, mucosal glycoprotein and glutathione levels and decreased the lipid peroxidation in gastric mucosa of diabetic rats. Its treatment also reversed the decrease in life span of gastric mucosal cells as indicated by decreased cell shedding in the gastric juice but found to have no effect on cell proliferation, indicating enhanced defensive status. E. jambolana extract was effective in reversing the delayed healing of gastric ulcer in diabetic rats near to the normal level. E. jambolana showed better ulcer healing effect than glibenclamide, because of its both antihyperglycemic and mucosal defensive actions. It could thus, be a better choice for treating gastric ulcers co-occurring with diabetes. PMID- 19810573 TI - Immunomodulatory activity of Acacia catechu. AB - The immunomodulatory effect of aqueous extract of Acacia catechu commonly known as Katha or Karangali was studied at two doses of 5 mg/kg and 50 mg/kg orally. The effect was studied in neutrophil adhesion test, mice lethality test, carbon clearance assay, cyclophosphamide induced neutropenia, serum immunoglobulin levels and the heamagglutination test. Acacia catechu extract showed an increase in the neutrophil adhesion to the nylon fibres, produced a significant increase in the phagocytic index and a significant protection against cyclophosphamide induced neutropenia indicating its effect on cell mediated immunity. On the other hand, Acacia catechu extract produced a significant increase in the serum immunoglobulin levels, increase in the haemagglutination titre values and decreased the mortality ratio in mice, suggesting its effect on the humoral arm of the immune system. From the above results, it was concluded that the aqueous extract of Acacia catechu has a significant effect on both cell mediated and humoral immunity. PMID- 19810574 TI - Homocysteine--risk factor for ischemic stroke? AB - The present study was undertaken to compare the homocysteine levels in patients of ischemic stroke with controls. Our study included 117 patients of ischemic stroke and 101 controls. The mean homocysteine levels in patients with ischemic stroke were 16.80 +/- 6.71 micromol/L while in controls it was 12.30 +/- 4.68 micromol/L, the difference being statistically significant (P < 0.01). The increased homocysteine levels in patients with ischemic stroke are independent of diabetes mellitus, age and sex. The homocysteine levels were higher in hypertensive subjects than non-hypertensive (P < 0.05). PMID- 19810575 TI - Protective effect of curcumin against kainic acid induced seizures and oxidative stress in rats. AB - The effect of curcumin, a dietary antioxidant was studied against kainic acid (KA)-induced seizures and on markers of oxidative stress. Rats were administered KA (10 mg/kg, ip) and observed for behavioral changes, incidence and latency of convulsions and mortality over four hours. The rats were thereafter sacrificed for estimation of oxidative stress parameters; malondialdehyde (MDA) and glutathione (GSH). Curcumin was administered 30 min before KA at doses of 50, 100 and 200 mg/kg, ip. KA induced long-lasting seizures and associated symptoms. The brain level of MDA was significantly (P < 0.05) raised after KA administration (536 +/- 44 nmol/g wet tissue) as compared to saline treated group (200 +/- 36 nmol/g wet tissue) and significantly decreased the levels of GSH. Pretreatment with curcumin (100 and 200 mg/kg, ip) significantly increased the latency of seizures (120 + 20 min and 11 5+/- 5.7 min respectively) as compared to the vehicle treated KA group. Curcumin (100 and 200 mg/ kg, ip) significantly prevented the increase in MDA levels and ameliorated the fall in glutathione. Curcumin at the dose of 50 mg/kg had no effect on any of oxidative stress parameters. The study reports the potential antiepileptic effect of antioxidant curcumin. PMID- 19810576 TI - Comparison of effects of olanzapine and risperidone on body mass index and blood sugar level in schizophrenic patients. AB - Schizophrenia is one of the most debilitating disorders with devastating effects on its victims and their families. Atypical antipsychotics (AAPs) because of their superior efficacy, reduced side effects, & better compliance, have rapidly become the mainstay of treatment. But, because of paucity of research & literature on the long-term metabolic side effect profile of these AAPs in Indian setup, this prospective study has been carried out to compare the effects of olanzapine & risperidone on body weight, body mass index, & blood sugar level in schizophrenic patients. Among 60 newly diagnosed DSM-IV patients of schizophrenia enrolled, it was observed that mean body weight & BMI were significantly increased from baseline to 6 & 12 weeks in both olanzapine (n = 30) & risperidone groups (n = 30) (P < 0.001). Also, mean blood sugar was found to be significantly elevated after 6 & 12 weeks of treatment with olanzapine (P < 0.001) but not in risperidone group. Thus, the present study underscores the need for baseline and six weekly monitoring of body weight and blood glucose in routine clinical practice with AAPs. PMID- 19810577 TI - High fat diet induces obesity in British Angora rabbit: a model for experimental obesity. AB - A reliable and cost-effective animal model for human obesity with its manifested disorders is yet to be established in the context of increased morbidity and mortality due to obesity and its related problems. Therefore, an attempt was made to produce obesity in locally available British Angora Rabbits (BAR) and examine the effect on metabolic and cardiovascular parameters. Adult male BARs weighing nearly 2 kg were randomly divided into two groups, one of the groups was fed with high fat diet (HFD) ad libitum for 10 weeks and the control group received standard normal rabbit chow for same period. Body weight, skinfold thickness, serum cholesterol, serum glucose and resting heart rate were measured before and after the dietary regimens. After 10 weeks, HFD group of rabbits demonstrated significant (P < 0.05) increase in body weight (+24%) and skinfold thickness (+37%). The gain in body weight was positively correlated to skinfold thickness (r = 0.61). Serum cholesterol, serum glucose and resting heart rate were also increased by 46%, 52% and 15%, respectively. Whereas no such increases in any of these parameters were observed in control group of rabbits. Our results suggest that obesity can be produced in BARs by feeding HFD. The obesity manifests with cardiovascular and metabolic changes. It is proposed that this may serve as a valid and reliable model of experimental obesity. PMID- 19810578 TI - Evaluation of antitussive activity of formulations with herbal extracts in sulphur dioxide (SO2) induced cough model in mice. AB - Cough is the most common symptom of respiratory diseases. When cough becomes serious, opioids are effective, but they have side effects like sedation, constipation, some addiction liability and also compromise the respiratory function. Therefore, there is need to have effective anti-tussive agent which do not have respiratory suppressant activity. The present study was carried out to evaluate anti-tussive activity of combination of herbal drugs as formulations in sulphur dioxide (SO2)-induced cough model in mice. Albino mice of either sex, weighing 25-30 g were divided into eight groups, (n = 6). Group 1 served as normal control, group 2 mice were given distilled water, group 3 was positive control and received codeine sulphate (10 mg/kg, p.o.) and group 4, 5, 6, 7 received coded 1 formulations 1, 2, 3 and 4 respectively at a dose of 0.3 ml/mice, orally, while group VIII was the vehicle control. Thirty minutes later, the mice were exposed to sulphur dioxide again for 45 sec. The mice were then placed in an observation chamber for counting of cough bouts, by two independent observers, for five minutes. All the formulations used showed significant antitussive activity in sulphur dioxide induced cough model. Thus, these formulations can prove to be useful for alleviating cough. PMID- 19810579 TI - Immediate effects of two relaxation techniques on healthy volunteers. AB - This controlled study compared immediate effects of two relaxation techniques on state anxiety and sustained attention in healthy subjects. 86 volunteers (56 men and 30 women) were divided into two groups: the first 43 volunteers (age range 18 to 64) practiced 20 minutes of yoga-based Deep Relaxation Technique (DRT), while the second group of 43 volunteers (same age range), practiced 20 minutes Supine Rest (SR). State anxiety was assessed using the State Trait Anxiety Inventory (STAI A-State), and sustained attention was assessed using the Six Letter Cancellation (SLC) and Digit Letter Substitution (DLS) tests. All tests were administered immediately before, and immediately after, practice. A significant reduction in State Anxiety score (P < 0.001) was observed for the group practicing DRT, but not for the group practicing SR. For the sustained attention tests, however, there were significant increases in scores by both DRT and SR groups (P < 0.001). The results suggest that both interventions improve attention, but that only DRT reduces State Anxiety. PMID- 19810580 TI - Study of plasma level of atorvastatin and its effect on lipid profile. AB - Atorvastatin, being one of the most commonly used antihyperlipidemic agents, is prescribed frequently by physicians all over the world but only a few data is available stating its effect in different ethnic population, more so from this part of India. The present study was designed focusing mainly on local population and was planned to determine plasma level of atorvastatin 10 mg and its effect on lipid profile in newly diagnosed hyperlipidemic patients attending Medical OPD of Shri Krishna Hospital, Karamsad (Gujarat). Study-I was carried out in 6 healthy volunteers to determine t(max) after single dose of atorvastatin 10 mg under fasting conditions, on the basis of which Study-II was conducted in 15 patients, collecting blood samples at a particular time (i.e t(max) of Study-I) after administration of atorvastatin 10 mg/day on day 1 as well as on the last day after 8 weeks of treatment. The plasma concentrations were determined by RP-HPLC system. Atorvastatin 10 mg/day for 8 weeks with a plasma level range (7.45, 12.08) ng/mL significantly (P < 0.05) reduced all the parameters of lipid profile from the study population. The mean decrease in HDL-C triggers a question on the effect of atorvastatin on HDL-C, which requires further study on a larger population of our country. PMID- 19810581 TI - Obesity attenuates formalin-induced tonic pain in British Angora rabbits. AB - Obesity is known to alter various physiological parameters including the pain sensitivity. There are conflicting reports on the pain sensitivity in obesity. In this context, the present study was aimed to investigate the tonic pain response in obese rabbit model. To achieve this aim, two groups of adult male British Angora rabbits were used. One of the groups was fed with standard rabbit chow and served as control. The other group was fed high fat diet (HFD) for 10 weeks to produce obesity. The standard formalin test was performed at the start and after 10 weeks of dietary regimen in both the groups. Timed behavioral responses (limping, elevation of paw, licking, biting, grooming etc.) were categorized and quantified with the help of standard pain rating scale. The total average pain rating score decreased significantly from 2.01 +/- 0.02 to 1.47 +/- 0.08 (P < 0.05) in HFD group after 10 weeks of dietary regimen, whereas there was no change in the control group. A significant negative correlation was observed between body weight and pain rating score in HFD group of rabbits (P < 0.05, r = -0.62). Results suggest that obesity attenuates the tonic pain responses induced by formalin in British Angora rabbits. PMID- 19810582 TI - Changes in hypothalamic and body temperatures during 24 hours in rats. AB - The available information on simultaneous assessment of body and hypothalamic temperatures in rats are lacking. In the present study these temperatures were recorded for 24 h, on three alternate days, in rats maintained at an ambient temperature of 26 +/- 1 degrees C. Hypothalamic temperature was significantly higher than body temperature during the night. In nocturnal rats the magnitude of circadian variation in hypothalamic temperature was higher than body temperature. Though maintained at different levels, both the temperatures showed closely associated diurnal changes throughout the 24 h period. PMID- 19810583 TI - Acute effects of formalin on pulmonary functions in gross anatomy laboratory. AB - Formaldehyde is a common indoor pollutant with irritant properties. The present study was undertaken in first year medical students to study the acute effects of it on their first exposure in gross anatomy laboratory. Twenty randomly selected subjects aged between 18 to 20 years and non smokers had undergone pulmonary function testing thrice. First being baseline values then immediately after their first dissection class (2 hrs.) and lastly 24 hrs after their first exposure. All the tests were performed on Siblemed 120 B portable spirometer with inbuilt computer programme under standard laboratory conditions. It was observed that FVC decreased in subjects immediately after their first exposure but reverted back to normal within 24 hrs. While all other lung function parameters remained unchanged, indicating some mild transient bronchoconstriction on acute exposure to formalin. The study needs to be extended with larger number of subjects. PMID- 19810584 TI - Effect of yoga-nidra on blood glucose level in diabetic patients. AB - Diabetes is a metabolic disorder, which has become a major health challenge worldwide. South East Asian countries have a highest burden of diabetes. In India the prevalence of diabetes is rising rapidly especially in the urban population because of increasing obesity and reduced physical activity. An objective of this study is to evaluate the effect of Yoga-Nidra on blood glucose level in diabetic patients. This study was conducted on 41, middle aged, type-2 diabetic patients, who were on oral hypoglycaemic. These patients were divided in to two groups: (a) 20 patients on oral hypoglycaemic with yoga-nidra, and (b) 21 were on oral hypoglycaemic alone. Yoga-nidra practiced for 30 minutes daily up to 90 days, parameters were recorded every. 30th day. Results of this study showed that most of the symptoms were subsided (P < 0.004, significant), and fall of mean blood glucose level was significant after 3-month of Yoga-nidra. This fall was 21.3 mg/dl, P < 0.0007, (from 159 +/- 12.27 to 137.7 +/- 23.15,) in fasting and 17.95 mg/dl, P = 0.02, (from 255.45 +/- 16.85 to 237.5 +/- 30.54) in post prandial glucose level. Results of this study suggest that subjects on Yoga-nidra with drug regimen had better control in their fluctuating blood glucose and symptoms associated with diabetes, compared to those were on oral hypoglycaemics alone. PMID- 19810585 TI - Comments on the short communication "Immediate effects of right and left nostril breathing on verbal and spatial scores" by Meesha Joshi and Shirley Telles in IJPP 2008; 52: 197-200. PMID- 19810586 TI - Nosocomial tuberculosis in the era of drug resistant tuberculosis. PMID- 19810587 TI - Utilization of RNTCP services in rural areas of Bellary District, Karnataka, by gender, age and distance from health centre. AB - BACKGROUND: Knowledge on utilization pattern of RNTCP shall provide important inputs towards its strengthening in rural areas. AIMS: To find out the utilization of RNTCP services by age, sex and distance from residence to designated microscopy centres and treating health centres. METHODS: The study was carried out in Sandur TU of Bellary District, Karnataka. Information on age, sex and residence of persons with pulmonary symptoms and detected new sputum smear positive cases during third quarter 2003 to second quarter 2004; and their treatment outcome was obtained from the respective RNTCP records. Age and sex distribution of out-patients was collected from OPD registers of one randomly selected DMC and its PHCs. RESULTS: A lesser number of males accessed the health care services. However, larger number of males with pulmonary symptoms and new sputum smear positive cases utilized RNTCP services than females in the ratio of 1.6:1 and 2.5:1 respectively. This was due to higher prevalence of persons with pulmonary symptoms and sputum positivity rate among males. Sputum positivity rates were also lower among the elderly. Male symptomatics and cases were on an average older than females. About 70% symptomatics and 53% cases resided at more than four kilometers from the respective DMCs and treating health centres. Treatment outcome was poorer among males with higher proportion of initial defaulters and among those residing at more than 20 kms. CONCLUSION: There is need to make health services available to the male working population at convenient hours and to be more vigilant to screen persons with pulmonary symptoms among the elderly. Collection of sputum specimen from eligible persons may be undertaken at PHCs which may later be transported to DMC. Supervision and motivation of treatment for male TB cases and those residing more than 20 kms from the treating health centres requires to be strengthened. PMID- 19810588 TI - Post-HAART tuberculosis in adults and adolescents with HIV in India: incidence, clinical and immunological profile. AB - BACKGROUND: Highly Active Antiretroviral Therapy (HAART) was introduced in National AIDS Control Programme in 2004 to reduce the morbidity and mortality among those affected with HIV/AIDS. Tuberculosis, being an important coinfection, its emergence/occurrence in post-HAART period has potential implications. OBJECTIVE: Primary objectives were to study the incidence of post-HAART tuberculosis in HIV patients and to identify the possible risk factors. It was also intended to understand the clinical and immunological profile of this important condition. METHODOLOGY: Eligible adults and adolescents with HIV disease enrolled on HAART at Government Hospital of Thoracic Medicine, Tambaram Sanatorium, Chennai, from April, 2004 to March, 2007, formed the study population. They were monitored and screened for the occurrence of tuberculosis after commencing HAART. Clinical details and immunological profile of these patients were analysed. RESULTS: Two hundred and sixty-two patients (5.1%) of 5099 patients followed-up for one to four years were found to have Post HAART TB with 100-person year risk of 2.83. Post HAART TB occurred predominantly in men (67.6%) and in 31-44 years age group (69.8%) with 100-person year risk being 3.26 and 2.83 respectively. Pulmonary, Extra-pulmonary and disseminated tuberculosis were found to occur in the frequencies of 78%, 16% and 6% respectively. A total of 144 patients (54.9%) developed tuberculosis within six months and this number increased to 202 (77%) by 12 months. 230 patients (87.7%) had base level CD4 cell count < 200 / mm3. CONCLUSION: Tuberculosis was found to occur pre-dominantly in adult male patients with HIV during the first year after the initiation of HAART. Significantly, occurrence of Post HAART TB remained almost the same (5%) among patients treated for TB prior to the initiation of HAART. PMID- 19810589 TI - Tuberculosis mortality trends in Delhi after implementation of RNTCP. AB - OBJECTIVES: To study the impact of Revised National TB Control Programme on mortality among tuberculosis patients in Delhi and to correlate mortality trends with programme indicators. METHODS: Record based evaluation of mortality trends from TB registers of all chest clinics of Delhi after implementation of Revised National TB Control Programme. RESULTS: The study showed a statistically significant decline in tuberculosis mortality among new smear positive cases after the implementation of Revised National TB Control Programme (z = 4.478 p < 0.05). The mortality among new smear negative and extra pulmonary cases also showed reduction, though not statistically significant. CONCLUSION: Mortality due to tuberculosis has been considerably reduced in Delhi over the years with the Revised National TB Control Programme implementation since 1997. PMID- 19810590 TI - Comparison of phenotypic and genotypic methods for pyrazinamide susceptibility testing. AB - OBJECTIVES: To evaluate Pyrazinamide (PZA) susceptibility results obtained by phenotypic MGIT 960 TB system against enzymatic Pyrazinamidase assay and genotypic pncA gene sequencing. To find the prevalence of infections caused by M. bovis in PZA resistant M. tuberculosis complex isolates. METHODS: 33 consecutive PZA resistant and 30 consecutive PZA susceptible isolates reported for PZA susceptibility testing by MGIT 960 TB system were included in this study. Presence of active pyrazinamidase enzyme was sought by using the Wayne assay. The pncA gene was amplified by PCR and then sequenced to screen mutations. All the PZA resistant isolates were further spoligotyped to identify M. bovis, if present. RESULTS: Of 33 PZA resistant strains by MGIT 960, 31 were Wayne assay negative and two were positive. Of the 30 susceptible PZA strains six were Wayne assay negative reporting false resistance. PncA gene sequencing revealed that 32 of the 33 MGIT PZA resistant isolates had diverse nucleotide changes scattered throughout the pncA gene (one isolate did not show any mutation). Of the 30 phenotypically susceptible isolates, 21 were wild types whilst nine isolates showed the presence of a silent mutation C-T at codon 195. Fifteen mutations found in this study has not been described earlier. Not a single isolate of M. bovis was detected among PZA resistant M. tuberculosis complex isolates. CONCLUSION: MGIT 960 showed better concordance with sequencing results in comparison with Wayne assay. In present study, a high proportion (85%) of MDR-TB isolates from patients receiving anti-TB treatment were found to be resistant to PZA. PMID- 19810591 TI - Status report on RNTCP. PMID- 19810592 TI - Two case reports of ultrasonography features in male genital tuberculosis. AB - Genitourinary tuberculosis is the common manifestation of extra-pulmonary tuberculosis. In the male genital tract, the epididymis followed by seminal vesicle, prostate, vas deferens and testis are commonly affected sites. Ultrasonography (USG) is the best imaging modality for the diagnosis of the diseases of male genital tract. We are presenting USG findings in two cases of male genital tuberculosis with involvement of the prostate, seminal vesicle, epididymis and vas deferens. PMID- 19810593 TI - Spina ventosa discharging tubercle bacilli--a case report. AB - Tubercular dactylitis is an unusual form of skeletal tuberculosis. Radiographic features of cystic expansion of the short tubular bones have led to the name of "Spina Ventosa" for tuberculosis dactylitis of the short bones. We report here a case of a Spina Ventosa in a 15 year-old female, who presented with complaint of draining sinus on the dorsum of the third digit of the left foot. PMID- 19810594 TI - Bcg induced mycobacterial spindle cell pseudotumor in an infant. AB - Mycobacterial Spindle cell Pseudotumor (MSP) is a rare complication of mycobacterial infection, especially the atypical variety. It is characterized by an exuberant spindle cell proliferation. This has been reported in the lymph nodes, skin, spleen, lungs, brain, etc. The incidence is higher in immuno compromised patients, especially those with acquired immunodeficiency syndrome. It is rare to encounter this lesion in infants. We report a case of MSP in the axillary lymph node of a 7-month-old infant, following Bacillus Calmette Guerin (BCG) vaccination due to Mycobacterium tuberculosis complex, which was proved by PCR. PMID- 19810595 TI - Outcome of directly observed treatment short course (DOTS) in a case of pulmonary tuberculosis with hyperuricemia. PMID- 19810596 TI - Growth of Cymbopogon citratus and Vetiveria zizanioides on Cu mine tailings amended with chicken manure and manure-soil mixtures: a pot scale study. AB - The Rakha Cu mines are located at East Singhbhum, Jharkhand, India and their activities ceased in 2001. The tailings (residue) were permanently stored in tailings ponds that require vegetation to reduce their impact on the environment. A pot scale study was conducted to evaluate the suitability of Cymbopogon citratus (DC.) Ex Nees and Vetiveria zizanioides (L) Nash for the reclamation of Cu tailings and to evaluate the effects of chicken manure and soil-manure mixtures on the revegetation of such tailings. Application of manure and soil manure mixtures resulted in significant increase in pH, EC, OC, CEC and nutritional status of Cu tailings. The environmentally available and DTPA extractable Cu and Ni concentration reduced in amended tailings, while Mn and Zn content increased significantly. Plants grown on amended tailings accumulated lesser Cu and Ni but higher Mn and Zn. Plant biomass increased proportionally to manure and soil-manure mixtures application rates. Lemon grass produced more biomass than vetiver grass in either of the amended tailings. From the pot experiment, it can be suggested that application of chicken manure @ 5% (w/w) and in combination with lemon grass, could be a viable option for reclamation (phytostabilization) of toxic tailings. PMID- 19810597 TI - Vetiver grass, Vetiveria zizanioides: a choice plant for phytoremediation of heavy metals and organic wastes. AB - Glasshouse and field studies showed that Vetiver grass can produce high biomass (>100t/ tha(-1) year(-1)) and highly tolerate extreme climatic variation such as prolonged drought, flood, submergence and temperatures (-15 degrees - 55 degrees C), soils high in acidity and alkalinity (pH 3.3-9.5), high levels of Al (85% saturation percentage), Mn (578 mg kg(-1)), soil salinity (ECse 47.5 dS m(-1)), sodicity (ESP 48%), anda wide range of heavy metals (As, Cd, Cr, Cu, Hg, Ni, Pb, Se, and Zn). Vetiver can accumulate heavy metals, particularly lead (shoot 0.4% and root 1%) and zinc (shoot and root 1%). The majority of heavy metals are accumulated in roots thus suitable for phytostabilization, and for phytoextraction with addition of chelating agents. Vetiver can also absorb and promote biodegradation of organic wastes (2,4,6-trinitroluene, phenol, ethidium bromide, benzo[a]pyrene, atrazine). Although Vetiver is not as effective as some other species in heavy metal accumulation, very few plants in the literature have a wide range of tolerance to extremely adverse conditions of climate and growing medium (soil, sand, and railings) combined into one plant as vetiver. All these special characteristics make vetiver a choice plant for phytoremediation of heavy metals and organic wastes. PMID- 19810598 TI - Effects of inoculation with arbuscular mycorrhizal fungi on maize grown in multi metal contaminated soils. AB - Pot culture experiments were established to determine the effects of colonization by arbuscular mycorrhizal fungi (AMF) (Glomus mosseae and G. sp) on maize (Zea mays L.) grown in Pb, Zn, and Cd complex contaminated soils. AMF and non-AMF inoculated maize were grown in sterilized substrates and subjected to different soil heavy metal (Pb, Zn, Cd) concentrations. The root and shoot biomasses of inoculated maize were significantly higher than those of non-inoculated maize. Pb, Zn, and Cd concentrations in roots were significantly higher than those in shoots in both the inoculated and non-inoculated maize, indicating the heavy metals mostly accumulated in the roots of maize. The translocation rates of Pb, Zn, and Cd from roots to shoots were not significantly difference between inoculated and non-inoculated maize. However, at high soil heavy metal concentrations, Pb, Zn, and Cd in the shoots and Pb in the roots of inoculated maize were significantly reduced by about 50% compared to the non-inoculated maize. These results indicated that AMF could promote maize growth and decrease the uptake of these heavy metals at higher soil concentrations, thus protecting their hosts from the toxicity of heavy metals in Pb, Zn, and Cd complex contaminated soils. PMID- 19810599 TI - Implantation of an atelocollagen sheet for the treatment of vocal fold scarring and sulcus vocalis. AB - OBJECTIVES: The management of vocal fold scarring and sulcus vocalis is challenging. These disorders are thought to be fibroplastic anomalies in the cover portion of the vocal fold that cause deterioration of the vibratory properties of the vocal fold mucosa. Histologic studies have revealed disorganization of extracellular matrix that needs to be addressed in the treatment of scarred vocal folds. Replacement of scar tissues with an appropriate implant may lead to regeneration of the vocal fold mucosa and its tissue properties. This retrospective case study examined the feasibility of using an atelocollagen sheet as a regenerative implant. METHODS: Six patients with a post cordectomy scar or sulcus vocalis underwent implantation of an atelocollagen sheet into the lamina propria of the vocal folds. The procedure consisted of elevation of a microflap, dissection and removal of scar tissue, implantation of the material, and wound closure. Vocal function was evaluated before and after surgery by stroboscopic examination and by aerodynamic and acoustic analyses. RESULTS: The postoperative changes of aerodynamic and acoustic parameters varied among patients; however, gradual improvement was seen in most cases over a year. Stroboscopic findings also revealed gradual improvement of vibratory properties in most cases. CONCLUSIONS: Implantation of an atelocollagen sheet may have restorative effects on vocal fold scarring and sulcus vocalis in terms of tissue properties and function of the mucosa. PMID- 19810600 TI - Piezoelectric surgical device in endoscopic sinus surgery: an initial clinical experience. AB - Revision endoscopic sinus surgery presents special challenges, such as the need to remove thick osteoneogenic bone close to the orbit or skull base. Currently available drills and microdebriders have shortcomings for this task, including risk to the underlying periorbita or dura. Recently, piezoelectric ultrasound technology has been used to remove bone while preserving adjacent soft tissue structures. This technology has been effective in several areas of the body, and it logically follows that it may have beneficial rhinologic applications. The purpose of this medical communication is to report an initial clinical experience with piezoelectric technology in revision sinus surgery. The piezoelectric surgical device was used during revision endoscopic sinus surgery in 14 patients in the author's practice from June 2006 to January 2009. All patients had an underlying bone component to their sinus condition, such as osteoneogenesis adjacent to the orbit or skull base. The piezoelectric surgical device performed successfully in removing osteoneogenic bone, and no complications were noted from its use. Piezoelectric surgical technology generates low-frequency ultrasound that dissects bone and appears to offer an option to mechanical drill instrumentation when used during endoscopic sinus surgery to address thick osteoneogenic bone. The clinical experience reported herein shows initial feasibility of the technology in selected cases of sinus surgery. On the basis of the favorable observations from this clinical experience, further exploration and discussion would appear to be valuable. PMID- 19810601 TI - Monitored anesthesia care with target-controlled infusion in vibroplasty. AB - OBJECTIVES: Correct positioning of a floating mass transducer during middle ear implant surgery is often problematic. With the use of monitored anesthesia care (MAC), however, deep sedation is maintained during surgery, followed by conscious sedation in which the patient can respond to test questions that investigate correct device position and function. The main aim of this study was to determine whether intraoperative audiometric assessment was feasible with MAC with target controlled infusion in vibroplasty. An additional aim was to determine whether MAC was sufficiently comfortable for patients during the procedure. METHODS: The study group comprised 8 patients who underwent vibroplasty under sedation. Before suturing, audiometric assessment was done by stimulating the external auditory processor with pure tones at 0.5, 1,2, and 4 kHz. Blood pressure, arterial oxygen saturation level, heart rate, and end tidal carbon dioxide level were monitored during the procedure and at awakening. RESULTS: Audiometric assessment was successfully completed in all 8 patients. The selected parameters indicated that no patient experienced pain or discomfort during surgery; the absence of discomfort was confirmed 1 to 2 hours after the operation by simple questioning. CONCLUSIONS: We found MAC to be an efficient and relatively safe technique for verifying the correct coupling of the floating mass transducer with the middle ear during vibroplasty. The patients were able to respond appropriately to questions and commands; moreover, none reported having experienced pain or discomfort during the operation. PMID- 19810602 TI - Endoscopic approach for pituitary surgery improves rhinologic outcomes. AB - OBJECTIVES: We hypothesized that the endoscopic approach to pituitary surgery improves rhinology-specific quality of life and has satisfactory tumor outcomes compared with the open approach. METHODS: Cases of pituitary surgery from the Department of Neurosurgery database included an inception cohort of all patients who had endoscopic procedures and consecutive patients who had open procedures between January 1998 and February 2008. The Sino-Nasal Outcome Test-22 was mailed. RESULTS: Since January 1998, 71 endoscopic and 122 open pituitary surgeries had been performed. The mean followup was longer for open procedures (49.3 months) than for endoscopic procedures (18.8 months). Recurrence was more common after open surgery (28.4%) than after endoscopic surgery (18.2%; p = 0.219). The most common diagnosis was macroadenoma (77.1% of endoscopic procedures and 93.4% of open procedures). The mean hospital stay was shorter for endoscopic procedures (4.1 days) than for open procedures (6.0 days; p <0.001). Of patients who presented with visual deterioration, 53.8% with endoscopic surgery and 46.7% with open surgery had improvement. Among patients with normal preoperative hormonal function, 27.5% of patients in the endoscopy group and 29.4% of patients in the open group required medication for more than 2 months after surgery. Complications occurred in 33.3% of endoscopic procedures and 43.4% of open procedures. Cerebrospinal fluid leaks were more common in the endoscopy group (p = 0.035), and diabetes insipidus lasting more than 30 days was more common in the open group (p = 0.017). The mean Sino-Nasal Outcome Test-22 score was lower for patients in the endoscopy group (20.4) than for those in the open group (23.2; p = 0.41). Patients in the endoscopy group had a significantly lower rhinology-specific mean score (6.5) than did patients in the open group (9.2; p = 0.03). CONCLUSIONS: The endoscopic approach to pituitary surgery offers tumor outcomes comparable to those of open surgery, with no greater incidence of complications and an improved rhinology-specific quality of life. PMID- 19810603 TI - Delayed cerebrospinal fluid leak following septoplasty. AB - We report on the clinical syndrome of delayed cerebrospinal fluid leak following septoplasty. We describe 2 such cases that we treated, and 1 other case described in the literature. A review of these 3 cases indicates a characteristic clinical and radiologic presentation and a consistent site of cerebrospinal fluid leak. All cases presented with cerebrospinal fluid leak 12 to 22 weeks after septoplasty and had slit-shaped dehiscences at the horizontal lamella of the cribriform plate. Endoscopic repair was successful in all. Delayed cerebrospinal fluid leaks may occur as a consequence of septal surgery. It is probable that uncontrolled twisting and rocking manipulations of the perpendicular plate of the ethmoid bone are transmitted to, and injure, the cribriform plate. PMID- 19810604 TI - Sphenopalatine and anterior ethmoidal artery ligation for severe epistaxis. AB - OBJECTIVES: We describe the surgical treatment of severe epistaxis and evaluate the recurrence of bleeding in a nonrandomized retrospective trial. METHODS: We performed a retrospective study comparing bilateral endoscopic ligation of the sphenopalatine artery alone (ELSPA) and bilateral endoscopic ligation of the sphenopalatine artery with concomitant bilateral external ligation of the anterior ethmoidal artery (ELSPEA) in the management of persistent epistaxis. Clinical and hematologic information, preoperative and surgical care, and short- and long-term outcomes were analyzed. The main outcome measure was recurrence of epistaxis in the short- and long-term follow-up periods. RESULTS: Forty-five patients were enrolled in the study. There were 20 patients in group A (ELSPA) and 25 in group B (ELSPEA). Three patients in group A and no patients in group B had long-term (more than 2 weeks after surgery) rebleeding. The difference between the two groups was not statistically significant (p > 0.05). CONCLUSIONS: We conclude that ELSPA and ELSPEA are effective, well-tolerated, reliable procedures if performed by an experienced surgeon. Their failure can be explained by anatomic lateral nasal wall variations and perioperative technical difficulties. They can be appropriate methods to treat severe recurrent epistaxis refractory to repeated nasal packing. PMID- 19810605 TI - Thyroid lymphosonography: a novel method for evaluating lymphatic drainage. AB - OBJECTIVES: We evaluated lymphosonography, or contrast-enhanced, ultrasonography (US)-guided sentinel lymph node (SLN) detection, as a technique for demonstrating the lymphatic drainage of the thyroid gland. METHODS: In this prospective animal study, four 50-kg Yorkshire swine underwent transcutaneous injection of a US contrast agent and methylene blue dye into the thyroid gland. Contrast-enhanced US was used to identify draining lymphatic channels and SLNs. Sentinel node biopsy was conducted. Subsequently, bilateral neck and upper mediastinal dissection was carried out. RESULTS: In 3 of 4 cases, a blue dye-positive and US contrast-positive SLN was identified. We identified SLNs in level IV in 2 cases. One case revealed 2 adjacent nodes in the superior mediastinum. In 1 case, a lymphatic channel was identified traveling into the mediastinum, but exposure of the SLN could not be obtained. No residual blue dye-positive or US contrast positive nodes were identified on subsequent dissection. CONCLUSIONS: Lymphosonography of the thyroid gland in a porcine model correlates well with blue dye-guided sentinel node biopsy and is technically feasible, although in some cases access to the SLN may be difficult. This technique could potentially enable a detailed analysis of thyroidal lymphatic drainage if applied to humans. PMID- 19810606 TI - Additional disease burden from hay fever and sinusitis accompanying asthma. AB - OBJECTIVES: We sought to determine the additional disease burden imparted by sinusitis and hay fever (allergic rhinitis) to patients with asthma. METHODS: Patients with a diagnosis of asthma, hay fever, or sinusitis were extracted from the National Health Interview Survey for the 1997 to 2006 adult sample. Disease groups consisting of patients with asthma alone, asthma + hay fever, asthma + sinusitis, and asthma + hay fever + sinusitis were assembled. Disease groups were then compared according to total health-care visits per year, emergency room visits per year, health-care spending per year, and number of workdays lost per year to determine the disease burden. RESULTS: We identified 11,813 patients (mean age, 45.5 years) who reported active asthma with or without hay fever or sinusitis comorbidity. Of these, 5,931 patients (50%) were identified with asthma alone, 1,134 (10%) with combined asthma + hay fever, 2,461 (21%) with asthma + sinusitis, and 2,287 (19%) with combined asthma + hay fever + sinusitis. Patients with asthma + sinusitis and those with asthma + sinusitis + hay fever had more total health-care visits and emergency room visits than did those with asthma alone (p <0.001). All three groups with comorbidities had higher healthcare expenditures than did the group with asthma alone (p < or = 0.002). Patients with asthma + sinusitis and those with asthma + hay fever + sinusitis missed more workdays than did patients in the group with asthma alone (10.0 and 13.1 versus 7.2, respectively; p <0.001). Comorbid hay fever alone did not increase workdays lost (6.6 days; p = 0.983). CONCLUSIONS: The additional disease burden of sinusitis on asthma is greater than that of hay fever. These data highlight the importance of identifying comorbid diagnoses with asthma. PMID- 19810607 TI - Clinical and electrophysiological characteristics of larynx in myasthenia gravis. AB - OBJECTIVES: We investigated the clinical and electrophysiological characteristics of the larynx in patients with myasthenia gravis (MG). METHODS: Thirty-two cases of MG were analyzed. The laryngeal behaviors and characteristics of laryngeal electromyography were evaluated, and voice assessment and repetitive nerve stimulation (RNS) tests were conducted. RESULTS: The initial symptoms of patients were ptosis and/or diplopia in 28, dysphagia and slurred speech in 3, and limb weakness in 1. Only 8 patients had slight hoarseness and vocal fatigue, and 4 patients had positive laryngeal signs. When compared with normal subjects, the patients with MG had worse results on the acoustic analysis of shimmer, their normalized noise energy was greater, and the harmonics-to-noise ratio and maximum phonation time were significantly lower. The RNS findings were positive in 28 patients and negative in 4 patients. The average (+/-SD) amplitude decrement was 31.9% +/- 19.1%. The mean number of involved laryngeal muscles was 2.22 +/- 1.35. Cricothyroid muscles were involved in 26 cases, thyroarytenoid muscles were involved in 14 cases, posterior cricoarytenoid muscles were involved in 6 cases, and lateral cricoarytenoid muscles were involved in 2 cases. CONCLUSIONS: Although few patients presented with laryngeal symptoms as their initial symptoms, most patients with MG exhibited asymmetry and abnormal findings on laryngeal electromyography. The RNS test for laryngeal muscles is a more sensitive indicator for the diagnosis of MG, especially in the cricothyroid muscle. PMID- 19810608 TI - Impact of distant metastasis in patients with cervical lymph node metastases from cancer of an unknown primary site. AB - OBJECTIVES: Treatment methods for patients with cervical cancer of an unknown primary site (CUP) are still under discussion. The purpose of this retrospective study was to analyze the oncological follow-up of 58 patients treated for cervical CUP. METHODS: From 1986 to 2006, 58 patients with cervical CUP were treated at the authors' institution. Treatment consisted of neck dissection alone in 8, irradiation or chemoradiation in 5, combined surgery and radiotherapy in 28, and surgery and radiochemotherapy in 17. RESULTS: The 3-year and 5-year overall survival rates for all patients were 52.9% and 40.9%, respectively. The 3 year and 5-year disease-specific survival rates were 50.9% and 39.7%, respectively. The 3-year and 5-year neck control rates were 73.7% and 67.3%, respectively. Neck dissection followed by radiochemotherapy was associated with the best 3-year and 5-year locoregional control rates. Extracapsular extension was a predictor of survival, but not of neck control. Distant metastases developed in about one third of all patients and were the most frequent cause of tumor-related death in cases of advanced neck disease. CONCLUSIONS: Despite the fact that regional control can be achieved in many cases, survival rates may be limited by distant metastasis, especially in patients with advanced neck disease. PMID- 19810609 TI - Locus for familial migrainous vertigo disease maps to chromosome 5q35. AB - OBJECTIVES: Migrainous vertigo (episodic vertigo associated with migraine) is sometimes inherited as an autosomal dominant trait. However, neither disease genes nor loci that might be responsible have been reported. We sought to map the genetic locus for familial migrainous vertigo in a 4-generation family and to define the progression of disease in this family. METHODS: We studied 23 members in a family in whom migrainous vertigo was inherited as an autosomal dominant trait. Clinical information obtained included case histories and results of otolaryngological, neurologic, audiometric, and imaging evaluations. Genome-wide linkage analysis was performed with Affymetrix Genechip Human Mapping 10K microarrays. Genotyping of family members' DNA with microsatellite markers was used to further assess candidate loci identified from the whole-genome scan. RESULTS: Of 23 family members, 10 suffered from migrainous vertigo beginning after 35 years of age. Migraine headaches usually preceded the onset of vertigo by 15 to 20 years. Longitudinal audiometric studies over 12 years showed stable, high-frequency sensorineural hearing loss consistent with presbycusis. Low frequency or fluctuating hearing loss was not observed. The results of vestibular testing and imaging studies were unremarkable. Genetic analysis defined a 12.0 MB interval on chromosome 5q35 between loci rs244895 and D5S2073 that contained the disease gene (logarithm of odds score, 4.21). CONCLUSIONS: We report the first locus for familial migrainous vertigo, which mapped to 5q35. PMID- 19810610 TI - Pepsin in nonacidic refluxate can damage hypopharyngeal epithelial cells. AB - OBJECTIVES: Studies using combined multichannel intraluminal impedance with pH monitoring reveal a role for nonacidic reflux in laryngopharyngeal symptoms and injury. We have discovered that pepsin is taken up by laryngeal epithelial cells by receptor-mediated endocytosis. This finding reveals a novel mechanism by which pepsin could cause cell damage, potentially even in nonacidic refluxate. The objective of this study was to determine whether pepsin, at pH 7.4 and thus in nonacidic refluxate, causes cell damage. METHODS: Cultured hypopharyngeal epithelial (FaDu) cells were exposed to human pepsin (0.1 mg/mL) at pH 7.4 for either 1 hour or 12 hours at 37 degrees C and analyzed by electron microscopy, cytotoxicity assay, and SuperArray. RESULTS: We report mitochondrial and Golgi complex damage in cells exposed to pepsin at neutral pH, observed by electron microscopy. We also report cell toxicity of pepsin at pH 7.4, measured by a cytotoxicity assay. Furthermore, using SuperArray, we found that pepsin at pH 7.4 significantly alters the expression levels of multiple genes implicated in stress and toxicity. CONCLUSIONS: These findings are perhaps the first to explain why many patients have symptoms and injury associated with nonacidic reflux, and could have important implications for the development of new therapies for reflux, such as pepsin receptor antagonists and/or irreversible inhibitors of peptic activity. PMID- 19810611 TI - Re: Preoperative and postoperative intracranial complications of acute mastoiditis. PMID- 19810612 TI - [From economic difficulties to social suffering of people living with HIV in Burkina Faso]. AB - A qualitative study conducted in 2006 shows that many people in Burkina Faso, living with HIV and receiving antiretroviral treatment (ART), have difficulties in meeting the expenses related to care. This anthropological analysis considers their perceptions, the causes and the social impact of these economic problems. This research is based on semi-structured interviews with 35 people living with HIV (PLWHIV) infection, contacted through HIV care programs in Ouagadougou, Bobo Dioulasso and Ouahigouya. After recording, transcribing and indexing the interviews, we analysed them. Even when they did not have to pay for treatment, most of the subjects faced economic problems dealing with follow-up care and monitoring, transportation, and other expenses, and these problems occur when PLWHIV have already faced the long, complex and expensive therapeutic itinerary before the test that diagnosed HIV infection. The new diet required by the treatment is also mentioned. Moreover, work problems often due to the disease (low output at work, inability to work, job loss) have already decreased patients' income by the time they get treatment. It is not easy for them to return to the level of resources they had before their disease, even if they can find a job. This financial exhaustion frequently leads to the exclusion of PLWHIV from their former solidarity networks, mostly because they can no longer participate in collective contributions or return what they were given when they were totally dependent. The analysis of these conversations shows the relevance of the concept of "social suffering" in describing the social effects of the economic problems that face many PLWHIV on ART, even when treatment is subsidized. PMID- 19810613 TI - [Maternal mortality in Libreville, Gabon: assessment and challenges]. AB - OBJECTIVE: To analyze the course over time of the rates and causes of maternal mortality in Libreville, Gabon. MATERIAL AND METHODS: This prospective descriptive study covers the 6-year period from January 1, 2001, to December 31, 2006. It was performed at the Libreville Hospital Center (LCH) maternity ward, the leading obstetrical care facility in the country. A maternal mortality registry began recording deaths on January 1, 2001. All maternal deaths are listed there, and a department meeting focusing on the death is organized after each. In this study, we analysed the epidemiologic characteristics, circumstances and cause of each death. RESULTS: In all, 136 women died. Their mean age was 25 +/- 5 years with a range of 16 to 41 years. Mean parity was 4 +/- 3, and ranged from 1 to 7. Most of the women were not employed (86%), but had at least some secondary schooling (88%). There were 126 cases (93%) of intrapartum deaths. They occurred more often at night (72% versus 28% during the day) and during the week (77% versus 23% on weekends). The maternal mortality rate was cut in half between 2000 and 2006 (p<0.005), dropping from 444 to 248 per 100 000 live births. Hemorrhages and eclampsia were the two principal causes of mortality. CONCLUSION: Faster access to care and improvement in the quality of emergency obstetrical care are essential conditions for reducing maternal mortality. PMID- 19810614 TI - [Prenatal diagnosis of trisomy 21: the Tunisian experience]. AB - Down syndrome (Trisomy 21) is the most common fetal chromosomal abnormality in humans. Its clinical signs are now well known. Methods for prenatal screening have advanced substantially in the past two decades. OBJECTIVE: To describe our experience with prenatal diagnosis of Down syndrome, including the indications, methods and results. MATERIAL AND METHODS: This retrospective study examined cases over a 4-year period. We adopted a sequential screening strategy for patients followed in our department since the beginning of their pregnancies after informed consent. We proposed first trimester ultrasound that measured nuchal translucency thickness and followed it with maternal serum screening. Some patients underwent screening during the second trimester or third trimester ultrasound. To assess the results, we studied the mothers' epidemiological characteristics and analysed the circumstances of prenatal diagnosis of trisomy 21 (T21). RESULTS/DISCUSSION: We identified 22 cases of T21 during the study period, for a total prevalence of 0.98 per thousand. The diagnosis was prenatal in 13 cases, mainly due to ultrasound signs. Of the 14 patients seen prenatally, only 8 were followed from early pregnancy. Five had enlarged nuchal translucency (> 95th percentile). Three had positive second trimester serum screening tests. One patient had amniocentesis planned because of her age (table 1). T21 was diagnosed in the second trimester in two cases and in the third trimester in three. The major morphological abnormalities observed were cardiac. We found an atrioventricular canal defect in four cases, and fetal hydrops in two cases (Table 2). The median gestational age at diagnosis of T21 in this study was 21 weeks. The diagnosis was missed in one patient followed throughout pregnancy in our unit. The median gestational age at termination of pregnancy was 22 weeks. Only one patient chose not to terminate the pregnancy. Her fetus, delivered at term, had no major pathologies. CONCLUSION: The establishment of a screening strategy for trisomy 21 in Tunisia is necessary to reduce handicaps. It should begin by expanding first-trimester ultrasound with nuchal translucency measurement. At the same time, serum marker testing should be offered to all patients. Routine amniocentesis for advanced maternal age should be avoided. PMID- 19810615 TI - [Digestive burns by simultaneous oral and rectal self-administration of ingestion sulphuric acid: an unusual mode of suicide]. AB - The authors report an unusual mode of suicide in two patients admitted to the Teaching Hospital of Yopougon. One 26-year-old woman swallowed sulphuric acid and then self-administered more of the same product rectally. She presented with abdominal pain, vomiting, bloody diarrhoea, without fever (T=37,9 degrees C). The abdomen did not appear to require surgery. Endoscopy showed erosive cesophagitis on day 5 (D5) and ulcerated bleeding inflammation of the entire colon and rectum on D26. She died on D30, during generalized convulsions. The second patient, a 31 year-old women, ingested sulphuric acid in the same way. She presented initially with diffuse abdominopelvic pains; clinical examination showed neither guarding nor contraction. Her temperature was 37,8 degrees C. Oesogastric endoscopy observed cesophagitis stage II B and III A and gastritis III A and III B. On D9, acute and generalized peritonitis was discovered, leading to surgical exploration. The patient died at the end of the operation. This unusual mode of suicide induces grave clinical presentations despite their apparent calm. The prognosis is worse because of the double localization of the lesions, the toxicity of the product, and the underestimation of its gravity. PMID- 19810616 TI - [Acanthamoeba keratitis: diagnosis to consider]. AB - INTRODUCTION: Acanthamoeba keratitis is a rare but severe disease that can cause blindness. The objective of this study is to call attention to its severity and conditions of development and to emphasize the importance of early treatment. CASE REPORT: We report a case of a 27-year-old woman. After wearing soft contact lenses for a month, she consulted for redness, pain and poor visual acuity in her left eye. The ophthalmological examination showed an epithelial corneal abscess measuring 2 mm in diameter in the left eye and very tiny epithelial corneal abscesses in the right eye. Microscopic examination and culture of samples from the contact lenses and the contact lens solution found Acanthamoeba spp. Rapid anti-amoeba treatment led to disappearance of the corneal abscesses and improvement of her vision. CONCLUSION: This case confirmed that the prognosis of Acanthamoeba keratitis depends on rapid diagnosis and early treatment. PMID- 19810617 TI - [Prevalence and polyparasitism of intestinal protozoa and spatial distribution of Entamoeba histolytica, E. dispar and Giardia intestinalis from pupils in the rural zone of Man in Cote d'Ivoire]. AB - Diseases caused by environmental contamination by micro-organisms, including intestinal helminths and protozoa, are prevalent in developing countries. According to some authors, their strong expansion in some zones of these countries is due primarily to favourable climatic conditions, combined with inadequate hygiene measures and cleaning and the generally low socio-economic level. Progress in disease control has resulted from new studies that improve our understanding of the epidemiology of helminthiases and from the availability of simple tools that are inexpensive and effective against these diseases (chemotherapy with albendazole and mebendazole). On the other hand, surprisingly few such studies have looked at intestinal protozoa, although the WHO reports that approximately 480 million individuals throughout the world are infested by amoebiasis caused by the protozoon Entamoeba histolytica and that 40,000-110,000 people die from it each year. Giardiasis, a cosmopolitan parasitosis, is due to another intestinal protozoon called Giardia intestinalis. To help develop a database on these parasites, we conducted a cross-sectional epidemiological survey in the Man region in western Cote d'Ivoire. Its objectives were to determine the prevalence of intestinal protozoa, to evaluate polyparasitism and to assess the spatial distribution of the pathogenic protozoal species, E. histolytica and G. intestinalis. Overall, 4466 stools samples taken from pupils aged 6 to 16 years of age at 57 different schools were analyzed under an optical microscope by the formol-ether stool concentration method, after preservation in sodium acetate-acetic acid-formalin (SAF). The most common protozoa species in this area were Endolimax nanus (83.8%) and E. coli (74.7%). The regional prevalence of G. intestinalis was 17.5% and of E. histolytica/E. dispar 11.3%. Both species were found in each of the 57 schools. The prevalence of E. histolytica/E. dispar exceeded 15% in six schools, and its spatial distribution was highly disparate. On the other hand, three large areas with substantial G. intestinalis transmission were identified. Polyparasitism of intestinal protozoa is very frequent in these areas; 80.2% of pupils carried at least 2 species. This work confirms the presence and breadth of protozoa species in the Man region. Understanding the spatial distribution of the principal areas where pathogenic protozoa species are transmitted may help to develop programmes for disease control that combine chemotherapy and preventive measures. PMID- 19810618 TI - [Causes of iron-deficiency anaemia in the internal medecine department of the national teaching hospital of Ouagadougou]. AB - This retrospective study in the internal medicine department of the national teaching hospital of Ouagadougou was conducted to identify the main causes of iron-deficiency anaemia. Among the 65 subjects meeting the inclusion and exclusion criteria, mean haemoglobin was 7.5 g/dl, with mean serum ferritin 8.9 microg/l among women and 15.5 microg/l among men. The most common cause was chronic blood loss, and hookworm was a major cause in 19.6% of cases. These results suggest the need for preventive measures against iron deficiency and for reinforcement of the fight against diseases producing fecal blood loss. PMID- 19810619 TI - [Causes of anaemia in African children aged 5 to 10 years]. AB - This study was conducted among two groups of 105 children in a hospital in Yaounde, Cameroon, to determine the causes of anaemia in children aged 5- 10 years-old. The study group included children presenting clinical evidence of anaemia while the control group was made of children without anaemia. Both sets of children were matched for neighbourhoods within the city of Yaounde. Each child had a complete physical examination, full blood count, haemoglobin electrophoresis, reticulocyte counts, and stool samples examined for parasites and hemoparasites. The mean haemoglobin level was 7.77 +/- 1.54 g/dL in the study group and 12.37 +/- 1.23 in the control group. The anaemia was mainly normocytic, normochromic and regenerative in the study group, usually associated with evidence of haemolysis. Patients with homozygous sickle cell disease were found only in the study group (n = 13; 12.3%). Malaria parasites were noted in 89 children (84.7%) in the study group, compared with 34 (32.4%) in the control group (p = 0.0004). Prevalence of intestinal parasites was similar in both groups (50.8%). Protein-calorie malnutrition was rare and was not associated with anemia. We conclude that both malaria and sickle cell anaemia play an important role in anaemia in children aged 5-10 years. PMID- 19810620 TI - [Aseptic osteonecrosis of the femoral head in children with sickle-cell disease]. AB - PURPOSE: To identify the epidemiological, clinical and therapeutic aspects of hip joint disorders in children with sickle cell disease, to point out the diagnostic problems, and to stress the necessity of early diagnosis for optimal outcome. MATERIAL AND METHODS: This retrospective study, conducted from January 1987 through December 2006, included children with at least one haemoglobin S gene and hospitalised for osteonecrosis of the femoral head. Ficat staging was used. RESULTS: The study included 14 children (12 boys and 2 girls) with a mean age of 14 years, all hospitalised and treated for osteonecrosis of the femoral head during the study period 8 SS and 6 SC. Osteonecrosis of the femoral head was diagnosed at Ficat stage 3 in 8 cases and at stage 4 in 6. Ten children (8 at stage 3 and 2 at stage 4) had orthopaedic treatment (continuous traction for 30 days and then a Thomas's splint for a mean 14 months). Outcome was good for 6 of the stage-3 cases (mean follow-up period 9 years) and poor for the other patients with purely orthopaedic treatment (mean follow-up period 11 years). Four children at stage 4 underwent surgery (varus osteotomy, immobilization in a cast for 8 weeks, and then Thomas's splint for a mean 12 months) with good results (mean follow-up period 10 years). CONCLUSION: Early diagnosis, assisted by computed tomography, magnetic resonance imaging or scintigraphy, makes it possible to provide better treatment and preserve hip function. PMID- 19810621 TI - [PubMed 2009. How to work with the new "advanced search" interface]. AB - PubMed/MEDLINE has recently changed the interface of its search function, especially the new web page called "Advanced Search", which combines on one page many of the functionalities and search options previous located at several different access points. We present here a tutorial to help French-speaking users work with this new interface. PMID- 19810622 TI - The Comarr Memorial Award for Distinguished Clinical Service: the legacy of A. Estin Comarr, MD. PMID- 19810623 TI - Bisphosphonate use in acute and chronic spinal cord injury: a systematic review. AB - BACKGROUND/OBJECTIVE: Bone density loss occurs rapidly after traumatic spinal cord injury (SCI) and is associated with low-energy fractures below the level of injury, commonly occurring around the knee. Bisphosphonates have been tested as potential agents to prevent bone loss after SCI, but no guidelines exist for clinical use of bisphosphonates in these patients. The objective of this study was to systematically review and evaluate evidence quality in studies of bisphosphonate use in patients with post-treatment follow-up of sublesional bone mineral density. METHODS: Literature search in MEDLINE/PubMed and ISI database using key words bisphosphonates, spinal cord injury, quadriplegia, paraplegia, and tetraplegia. RESULTS: The search identified 6 experimental studies and 1 quasi-experimental study of bisphosphonate therapy in patients with acute and chronic SCI. The studies were small and of fair or poor quality, and none included fracture outcomes. Mild attenuation of bone density loss with acute administration of bisphosphonates after SCI was found at some measurement sites but was not always maintained during follow-up. CONCLUSIONS: Data were insufficient to recommend routine use of bisphosphonates for fracture prevention in these patients. Current studies are limited by heterogeneity of patient populations and outcome measures. Uniform bone density measurement sites with rigorous quality control and compliance monitoring are needed to improve reliability of outcomes. Future studies should address specific populations (acute or chronic SCI) and should assess fracture outcomes. PMID- 19810625 TI - Association of mode of locomotion and independence in locomotion with long-term outcomes after spinal cord injury. AB - BACKGROUND/OBJECTIVE: To explore the association of mode of locomotion (ambulation vs wheelchair use) and independence in locomotion (independent vs require assistance) with health, participation, and subjective well-being (SWB) after spinal cord injury (SCI). RESEARCH DESIGN: Secondary analysis was conducted on survey data collected from 2 rehabilitation hospitals in the Midwest and a specialty hospital in the southeastern United States. The 1,493 participants were a minimum of 18 years of age and had traumatic SCI of at least 1 year duration at enrollment. MAIN OUTCOME MEASURES: Three sets of outcome measures were used: SWB, participation, and health. SWB was measured by 8 scales and a measure of depressive symptoms, participation by 3 items, health by general health ratings, days in poor health, hospitalizations, and treatments. RESULTS: Small but significant associations were observed between independence in locomotion and every outcome. Ambulation was associated with greater participation but a mixed pattern of favorable and unfavorable health and SWB outcomes. Supplemental analyses were conducted on those who ambulated but who were dependent on others to do so (n = 117), because this group reported poor outcomes in several areas. Individuals who were independent in wheelchair use reported substantially better outcomes than nonwheelchair users and those dependent on others in wheelchair use. CONCLUSIONS: Although ambulation is often a recovery goal, individuals with SCI who ambulate do not uniformly report better outcomes than wheelchair users, and those who depend on others for assistance with ambulation may experience a unique set of problems. PMID- 19810624 TI - Measurement of sexual functioning after spinal cord injury: preferred instruments. AB - BACKGROUND/OBJECTIVE: To determine the utility of certain instruments to assess sexuality and fertility after SCI, an expert panel identified key areas to study and evaluated available instruments. These were rated according to certain predefined criteria. METHODS: The authors divided sexual issues into male and female sexual function, male reproductive function, and female reproductive function. The instruments that have been used most frequently to measure these aspects of sexual function over the past 5 years were identified by expert consensus. Finally, these instruments were subjected to a critical review. RESULTS: The Female Sexual Function Index (FSFI), measurement of vaginal pulse amplitude (VPA), the International Index of Erectile Function (IIEF), and the measurement of ejaculatory function and semen quality were considered appropriate measures to assess sexual responses and reproductive function after SCI. There were no measures identified to assess female reproductive function. CONCLUSIONS: For clinical trials aiming to improve sexual function after SCI, the FSFI or the IIEF is currently preferred. Although VPA is an appropriate means to assess female sexual responses, it is only useful for laboratory studies and is too invasive for use in clinical trials. For assessment of male fertility potential, assessment of ejaculatory capacity and semen analysis are recommended. PMID- 19810626 TI - The SCIRehab Project: classification and quantification of spinal cord injury rehabilitation treatments. Preface. PMID- 19810628 TI - Clinical taxonomy development and application in spinal cord injury research: the SCIRehab Project. AB - BACKGROUND/OBJECTIVE: Applying practice-based evidence research methodology to spinal cord injury (SCI) rehabilitation requires taxonomy (typology or classification) of rehabilitation interventions provided by every discipline contributing to SCI rehabilitation. The rehabilitation field currently lacks such taxonomy. METHODS: SCIRehab project researchers and clinicians representing 7 rehabilitation disciplines from 6 US inpatient SCI rehabilitation facilities worked in discipline groups during 2 face-to-face meetings and weekly discipline specific teleconferences for 9 months to identify key contributions of each discipline to SCI rehabilitation and to develop a classification of treatment interventions used by each discipline. These clinician groups were charged with designing documentation systems that collected enough details to describe treatment adequately while not imposing an unrealistic data collection burden on clinicians. Completed documentation systems were programmed onto handheld personal digital assistants (PDAs) to facilitate data entry by clinicians at the point of care. RESULTS: Seven discipline-specific SCI rehabilitation taxonomies were developed that describe and quantify intervention activities (major categories of treatment offered by the discipline) and the activity-specific details (variables deemed important to fully describe the interventional process). Much treatment information is unique to each discipline; some is common across disciplines. CONCLUSIONS: The taxonomies provide a format with which clinicians document actual interventions performed with or for patients. The SCIRehab project has developed the first comprehensive multidisciplinary taxonomy for describing the details of the SCI rehabilitation process and designed a PDA based documentation system based on that taxonomy that allows clinicians to describe the specifics of their interactions with their patients. PMID- 19810627 TI - New approach to study the contents and outcomes of spinal cord injury rehabilitation: the SCIRehab Project. AB - BACKGROUND/OBJECTIVE: Limited research evidence is available to show the effectiveness of the many specific interventions provided in spinal cord injury (SCI) rehabilitation; what is available typically focuses on effects of the full rehabilitation package but not specific therapy interventions, medical procedures, patient education, or counseling. Given the problems of conducting randomized controlled trials (RCTs) in rehabilitation, practice-based evidence (PBE) research has been suggested as an alternative methodology for identifying which rehabilitation interventions are associated most strongly with positive outcomes, after controlling for patient differences. Using the PBE research methodology, the SCIRehab project attempts to "open the black box" of acute SCI rehabilitation, provide detailed information on treatments delivered by all rehabilitation disciplines, and contribute to outcomes-based guidelines for clinical decision-making. METHODS: The SCIRehab project includes 1,500 patients with acute SCI, consecutively admitted to 1 of 6 US inpatient rehabilitation facilities. Details of the rehabilitation process are captured by clinicians from multiple disciplines documenting their interventions in handheld personal digital assistants after sessions with their patients. Outcome data are abstracted from medical records (clinical outcomes data) and obtained from patient interviews at 6 and 12 months after injury. Extensive patient, injury, and other treatment characteristics are abstracted from medical records. SCIRehab is the first research project to collect detailed information on individual interventions offered by the full rehabilitation team. RESULTS: SCIRehab is the first research project to collect detailed information on individual interventions offered by the full rehabilitation team. These findings are presented in a series of 9 articles. CONCLUSIONS: To date, SCIRehab's major contribution is a system for categorizing specific contributions of each discipline and a technology for documenting that detail. After data collection is complete, future manuscripts will relate those process elements to outcomes. The SCIRehab Project is an important step toward establishing outcomes-based guidelines for SCI rehabilitation. PMID- 19810629 TI - SCIRehab Project series: the physical therapy taxonomy. AB - BACKGROUND/OBJECTIVE: Outcomes research is in need of a classification system of physical therapy (PT) interventions for acute traumatic spinal cord injury (SCI) rehabilitation in the United States. The objective of this study was to describe a taxonomy (system to categorize and classify interventions) to examine the effects of PT interventions on rehabilitation outcomes. METHODS: The SCIRehab study uses the rigorous observational practice-based evidence methodology to examine current treatment processes without changing existing practice. PT clinicians and researchers from 6 centers developed a taxonomy to describe details of each PT session. RESULTS: The PT taxonomy consists of 19 treatment activities (eg, bed mobility, transfers, wheelchair mobility, strengthening and stretching exercises) and supplementary information to describe the associated therapeutic interventions. Details that focus on patient assistance needs and family involvement are included as additional descriptors to help to describe and justify PT activity selection. Time spent on each activity is used as the measure of intensity. CONCLUSION: The detailed PT taxonomy documentation process, which offers efficiency in data collection, is being used for all PT sessions with 1,500 patients with acute traumatic SCI at the 6 participating centers. It might be the first attempt to document the many details of the PT rehabilitation process for patients with SCI in the United States. PMID- 19810630 TI - SCIRehab Project series: the occupational therapy taxonomy. AB - BACKGROUND/OBJECTIVE: Lack of a classification system for occupational therapy (OT) rehabilitation interventions for traumatic spinal cord injury (SCI) rehabilitation in the United States makes conducting outcomes research difficult. This article describes an OT SCI rehabilitation taxonomy (system to categorize and classify treatments). METHODS: OT clinicians and researchers from 6 SCI rehabilitation centers developed a taxonomy to describe details of each OT session. This effort is part of the SCIRehab study, which uses the practice-based evidence, observational research methodology to examine current treatment processes without changing existing practice. RESULTS: The OT taxonomy consists of 26 OT activities (eg, training on activities of daily living, communication, home management skills, wheelchair mobility, bed mobility, transfers, balance, strengthening, stretching, equipment evaluation, and community reintegration). Time spent on each activity is documented along with therapeutic interventions used to facilitate the activity. Treatment descriptions are enhanced further with identification of assistance needs, patient direction of care, and family involvement, which help to describe and guide OT activity selection. The OT taxonomy documentation process includes all OT rehabilitation interventions for patients with SCI while maintaining efficiency in data collection. CONCLUSION: The electronic documentation system is being used at 6 centers for all OT sessions with 1,500 patients with acute traumatic SCI. It is the largest known attempt to document details of the comprehensive OT rehabilitation process for patients with SCI in the United States. PMID- 19810631 TI - SCIRehab Project series: the therapeutic recreation taxonomy. AB - BACKGROUND/OBJECTIVE: Outcomes research of therapeutic recreation (TR) activities and interventions for spinal cord injury (SCI) rehabilitation is made more difficult by a lack of uniform descriptions and the absence of a formal treatments classification system (taxonomy). The objective of this study was to describe a taxonomy developed by Certified Therapeutic Recreation Specialists. METHODS: TR lead clinicians and researchers from 6 SCI rehabilitation centers developed a TR documentation system to describe the details of each TR session involving patients with SCI enrolled in the SCIRehab study. The SCIRehab study uses the practice-based evidence methodology, a rigorous observational methodology that examines current practice without introducing additional treatments, to capture details of each TR session for 1,500 SCI rehabilitation patients at 6 US inpatient SCI rehabilitation facilities. This may be the first attempt to document the many details of the TR rehabilitation process for patients with SCI. RESULTS: The TR taxonomy consists of 6 activities (eg, leisure education and counseling, outings, and leisure skill work in center) and activity specific interventions, as well as time spent on each activity. Activity descriptions are enhanced with additional details that focus on assistance needs for each activity, patient ability to direct care, and patient/family involvement, which may help to determine TR activity selection. CONCLUSION: Development and application of a TR taxonomy, which is comprehensive for patients with SCI and efficient to use, are feasible despite significantly different TR programs at the 6 SCIRehab centers. PMID- 19810632 TI - SCIRehab Project series: the speech language pathology taxonomy. AB - INTRODUCTION: Outcomes research for speech language pathology (SLP) interventions for acute traumatic spinal cord injury (SSC) rehabilitation in the US is difficult because of the lack of a treatment classification system (taxonomy). OBJECTIVE: To describe a taxonomy developed by speech language pathologists (SLPs) to examine the effects of SLP interventions on SCI rehabilitation outcomes. METHODS: The SCIRehab study uses practice-based evidence, a rigorous observational methodology that examines treatment processes without specifying or requiring specific therapeutic interventions. Speech language pathology lead clinicians and researchers at 6 US SCI centers developed a detailed SLP taxonomy documentation process that is comprehensive of SLP interventions for patients with SCI. RESULTS: The SLP taxonomy consists of 7 intervention categories that address deficits (speech production for patients with artificial airway, motor speech and voice, swallowing, cognitive-communication, and communication) and the associated exercises and tasks that patients perform. Time is recorded for each category, and supplementary information focuses on cueing needs and family involvement that helps to describe and guide intervention selection. The SCIRehab project is enrolling 1,500 patients with acute traumatic SCI at 6 inpatient rehabilitation facilities. CONCLUSIONS: Speech language pathology taxonomy information is being captured for the SCIRehab patients who are referred for SLP services; this may be the first attempt to document the many details of the SLP rehabilitation process for patients with SCI in the US. PMID- 19810633 TI - SCIRehab Project series: the psychology taxonomy. AB - CONTEXT: The integration of psychologists as members of the rehabilitation team has occurred in conjunction with the evolution and adoption of interdisciplinary teams as the standard of care in spinal cord injury (SCI) rehabilitation. Although the value of psychological services during rehabilitation is endorsed widely, specific interventions and their association with patient outcomes have not been examined adequately. OBJECTIVE: To address this shortcoming, psychologists from 6 SCI centers collaborated to develop a psychology intervention taxonomy and documentation framework. METHODS: Utilizing an interactive process, the lead psychologists from 6 centers compiled an inclusive list of patient characteristics assessed and interventions delivered in routine psychological practice at the participating rehabilitation facilities. These were systematically grouped, defined, and compared. RESULTS: The resulting taxonomy became the basis of a documentation framework utilized by psychologists for the study. The psychology taxonomy includes 4 major clinical categories (assessment, psychotherapeutic interventions, psychoeducational interventions, and consultation) with 5 to 10 specific activities in each category. CONCLUSIONS: Examination of psychological interventions and their potential association with positive outcomes for persons who sustain SCI requires the development of a taxonomy. Results of these efforts illustrate similarities and differences in psychological practice among SCI centers and offer the opportunity to blend research and clinical practice in an innovative approach to evidence-based practice improvement. The established taxonomy provides a basic framework for future studies on the effect of psychological interventions. PMID- 19810634 TI - SCIRehab Project series: the supplemental nursing taxonomy. AB - CONTEXT: Spinal cord injury (SCI) rehabilitation nurses document the occurrence of educational and care management efforts in traditional nursing documentation methods but not the intensity (or dose) of such interactions. This article describes a process to capture these nursing interventions. METHODS: Nurses at 6 US inpatient SCI centers used 2 in-person meetings and weekly telephone calls over 9 months to develop a taxonomy of nursing patient education efforts and care management. RESULTS: This was subsequently incorporated into a point-of-care documentation system and used to capture details of nursing care for 1,500 SCI rehabilitation patients enrolled in the SCIRehab study. The taxonomy consists of 10 education and 3 care management categories. The point-of-care system includes time spent on each category along with an indication of whether the patient and/or family received the education/care management. In addition, a subjective measure of patient participation in nursing activities is included. CONCLUSIONS: Creation of a SCI rehabilitation nursing taxonomy is feasible, and its use has had an impact on nursing practice. It also has implications for future clinical documentation, because greater accuracy and details of patient education and care management will be a permanent practice in the participating systems at the conclusion of the study. PMID- 19810635 TI - SCIRehab Project series: the social work/case management taxonomy. AB - CONTEXT: Social work and case management (SW/CM) are integral components of acute inpatient spinal cord injury (SCI) rehabilitation. However, evidence is sparse regarding the impact of SW/CM interventions on outcomes. To advance research on SW/CM clinical practice in SCI rehabilitation, SW/CM providers and researchers first must have standard classifications for SW/CM interventions. BACKGROUND/OBJECTIVE: To develop a taxonomy (classification) of the various interventions and services that comprise SW/CM. METHODS: A group of SW/CM clinicians compiled a list of activities performed as routine practice at the participating rehabilitation facilities. These activities were grouped and defined systematically. RESULTS: The resulting taxonomy includes 8 major activity topics (financial planning, discharge planning, discharge services, supportive counseling, information about and referral to peer/advocacy groups, education about SCI and other relevant topics, information about and referral to community/in-house services, and team conferences), which were further stratified into specific content areas. Interactions with the patient, family, or other team members and resources, along with descriptions of the interactions that are applicable to each of the 8 activity topics, were included as well. CONCLUSION: An intervention taxonomy is required to study the SW/CM interventions and the potential association with positive rehabilitation outcomes for patients with SCI. The SW/CM taxonomy developed for the SCIRehab project, which will be used with 1,500 patients admitted to 6 SCIRehab centers over 2.5 years, will provide an infrastructure for such research. PMID- 19810636 TI - Fatal fever of unknown origin in acute cervical spinal cord injury: five cases. AB - BACKGROUND/OBJECTIVE: Patients with traumatic upper thoracic and cervical spinal cord injuries are at increased risk for the development of autonomic dysfunction, including thermodysregulation. Thermoregulation is identified as an autonomic function, although the exact mechanisms of thermodysregulation have not been completely recognized. Quad fever is a hyperthermic thermoregulatory disorder that occurs in people with acute cervical and upper thoracic spinal cord injuries. First described in 1982, it has not been widely discussed in the literature. METHODS: Case reports of 5 patients with cervical spinal cord injury (SCI). RESULTS: Five of 18 patients (28%) with acute cervical SCI who were admitted during a 1-year period had fatal complications caused by persistent hyperthermia of unknown origin. CONCLUSIONS: Patients with acute traumatic cervical and upper thoracic SCI are at risk for thermoregulatory dysfunction. Changes in the hypothalamic axis may be implicated, especially in the light of modification in hypothalamic afferent nerves, but this hypothesis has not yet been explored. Thermodysregulation may be an early sign of autonomic dysfunction. A comprehensive guideline is needed for the management of elevated body temperature in critically ill patients with cervical SCI, because this condition may be fatal. PMID- 19810637 TI - Anterior spinal artery syndrome in a girl with Down syndrome: case report and literature review. AB - BACKGROUND/OBJECTIVE: Anterior spinal artery syndrome is an extremely rare cause of acute ischemic cord infarction in children. It is caused by hypoperfusion of the anterior spinal artery, leading to ischemia in the anterior two thirds of the spinal cord. The presentation is usually with an acute and painful myelopathy with impaired bladder and bowel control. Pain and temperature sensation below the lesion are lost, whereas vibration and position sense is intact because of the preservation of the posterior columns. METHODS: Case report. RESULTS: A 16-year old girl with Down syndrome presented with urinary retention and acute complete flaccid paralysis of the legs with absent deep tendon and abdominal reflexes. Magnetic resonance imaging showed a signal abnormality in the anterior half of the thoracic cord from T5 to T12, consistent with anterior spinal artery infarction. CONCLUSIONS: Pediatricians should consider anterior spinal artery syndrome in the child who presents with acute, painful myelopathy. We summarize the etiology, neurological findings and outcomes of 19 children found in the literature with anterior spinal artery syndrome. PMID- 19810638 TI - Free lunches and influence. PMID- 19810639 TI - Moral courage. PMID- 19810640 TI - Smile savers system may make a dentist's office smile. PMID- 19810641 TI - Specialist's failure to follow up leads to litigation. PMID- 19810642 TI - The business of dentistry. AB - There are many reasons why someone chooses to become a dentist: a family member was in the profession, they have a proclivity for the sciences of health care, or they like the autonomy of having their own practice. Mark Gonthier, assistant dean for admissions of Tufts University School of Dental Medicine, offered this description of the ideal dental school applicant, "We are looking for mature, well-rounded students capable of handling the rigor of the basic sciences curriculum and are equally adept and committed to providing quality comprehensive patient oral health care" Few dentists will report that they chose their profession because they wanted to run a small retail business. PMID- 19810643 TI - The legal business of dentistry. AB - Upon graduation and licensure, most dentists anticipate going into the profession of providing dental heath care to patients in an office or clinic setting. The profession is also the business of dentistry. Failure to appreciate documentation requirements for the business of dentistry can result in legal battles that are time-consuming and emotionally draining. This article provides an introduction, issue spotting, and tips to avoid those legal battles. PMID- 19810644 TI - Practical employment strategies: win-win solutions for dentists and their employees. AB - The profession of dentistry is also the business of dentistry and subject to the same employment laws as any other small business. Wrongful termination, discrimination, and harassment claims are on the rise in large part because dental employers often do not understand their basic employment law obligations. Good intentions do not offset inadequate office policies and documentation. Implementing, communicating, and evenly enforcing employment policies will reduce claims and promote office moral and productivity. PMID- 19810645 TI - Making standard of caring part of the standard of care. AB - Effective risk management focuses on consistent patient communication to inform patients of the benefits of dental health and the breadth of alternative treatments. When education is effective, it helps patients develop new understandings of health and disease. New understandings make appropriate treatment choices possible, and those choices reduce the chances of legal action and contribute to the health of the patient as well as the health of the practice. PMID- 19810646 TI - Complication or substandard care? Risks of inadequate implant training. AB - The use of dental implants has increased substantially over the past decade, as have the number of implant-related complications. One of the contributing factors is inadequate training in the prevention, recognition, and treatment of complications. Comprehensive training on implant surgical procedures that includes patient selection, risk management, and complications allows dentists to incorporate implant placement into their practices with less risk to patients and less risk of serious legal consequences. PMID- 19810647 TI - Cone beam CT--anatomic assessment and legal issues: the new standards of care. AB - Until the recent introduction of cone beam computed tomography scanners, standard 2-D imaging provided a moderate contribution to overall treatment planning when considering the diagnostic potential, costs of study, and risks to the patient. Cone beam computed tomography-dedicated maxillofacial imaging scanners provide broader imaging tools for anatomic assessment and have become widely available. This article discusses the uses and benefits of 3-D imaging, as well as the impact on the standard of care. PMID- 19810648 TI - H2Ohhhhhhhhhh. PMID- 19810649 TI - What can we really expect from healthcare reform? AB - Sorting through all of the charges and countercharges that have been raised against the various healthcare reform bills is a challenge as we enter what is likely to be the final critical period that will determine the fate of healthcare legislation in 2009. It's probably not surprising that this is where we find ourselves. PMID- 19810650 TI - Laboratory and the art of enterprise integration. AB - Many healthcare organizations are weighing the advantages of investing in a new laboratory information system (LIS) that promises greater accuracy and efficiency against the challenges of replacing the organization's existing LIS system. A recent KLAS survey considers the ROI of replacing an LIS with a next-generation solution, and whether providers are trending toward single-vendor integration or best-of-breed feature sets. Almost 70 percent of the 266 providers interviewed believe there is no "best" LIS. PMID- 19810651 TI - The value of value-based purchasing. AB - In the first four years of the Hospital Quality Incentive Demonstration (HQID) project operated by the Centers for Medicare & Medicaid Improvement (CMS) and Premier, Inc., HQID hospitals improved their composite quality scores by 17.2 percent across five clinical areas. If every hospital were able to replicate this level of performance, an estimated $4.5 billion and 70,000 lives would be saved each year, according to CMS and Premier. PMID- 19810652 TI - David Walker. Reduce costs first, then expand coverage. Interview by Richard L. Clarke. PMID- 19810653 TI - Do you know the fair market value of quality? AB - To develop a physician compensation package that includes fair-market-value incentive payments for their efforts to improve healthcare quality, a hospital first needs to: Evaluate current market data on quality incentive payments. Be familiar with the existing regulatory guidelines related to paying for quality. Understand the requirements for complying with the regulations. PMID- 19810654 TI - The business case for quality. AB - Among the lessons St. Joseph Medical Center learned in implementing a diabetes care management program were that: There is a sound business case for quality with such a program. A sound business plan based on a track record of accomplishments by other organizations is key to gaining clinician buy-in. Deploying dwindling resources to simultaneously improve care and the organization's bottom line requires collaboration between clinicians and finance. PMID- 19810655 TI - Using business intelligence to improve performance. AB - Cleveland Clinic's enterprise performance management program offers proof that comparisons of actual performance against strategic objectives can enable healthcare organization to achieve rapid organizational change. Here are four lessons Cleveland Clinic learned from this initiative: Align performance metrics with strategic initiatives. Structure dashboards for the CEO. Link performance to annual reviews. Customize dashboard views to the specific user. PMID- 19810656 TI - The impact of comparative effectiveness on U.S. healthcare providers. AB - Comparative effectiveness research (CER) has the potential to improve health care and reduce costs, yet it is controversial. Consumers need to learn about CER to understand its benefits and limitations. Providers need to manage changing evidence about what constitutes best practices. PMID- 19810657 TI - What is the hospital industry's exposure from the ARS collapse? AB - Auction rate securities (ARSs) were introduced 1988 as a means to allow tax exempt/municipal issuers to enjoy the advantage of short-term interest rates on long-term debt issues. The recent collapse in the ARS market caused many hospitals and healthcare systems to be confronted with a much higher cost of debt. In the context of the larger economic crisis facing our nation and healthcare systems, the problems in the ARS market may have only exacerbated the difficulties many U.S. hospitals face in accessing cash needed to fund essential capital projects. PMID- 19810658 TI - Depreciating and stating the value of hospital buildings what you need to know. AB - Healthcare financial executives of not-for-profit hospitals may be overdepreciating and understating the value of the hospital building on their financial statements. Changing the remaining lives of assets and their depreciation will help enhance the bottom line for many organizations. Ensuring that they are correctly stating the investment value of their assets is one way CFOs can have a positive impact on their organization's bottom line in a tough economy. PMID- 19810659 TI - 5 ways to keep your physicians engaged. PMID- 19810660 TI - How should a hospital expand its cancer services? PMID- 19810661 TI - Replacement hospitals: variance in operating margin compared with the national average. PMID- 19810662 TI - Persistent subepithelial haze in thin-flap LASIK. AB - PURPOSE: To report persistent subepithelial haze in two patients following femtosecond LASIK associated with creation of a thin flap. METHODS: Subepithelial haze was assessed by slit-lamp photography, high-resolution Scheimpflug imaging, and corneal confocal microscopy. RESULTS: Two patients showed distinct subepithelial haze and reduced corrected distance visual acuity at 3 months after LASIK with a think-flap generated by a femtosecond laser. The extent of haze was documented, and the haze was treated topically with steroids up to 12 weeks. The haze was localized approximately 20 to 40 microm below Bowman's layer and dissolved slowly during the 2 months of treatment. At 6 months after surgery, uncorrected visual acuity was 20/20. CONCLUSIONS: Subepithelial haze formation represents a new potential complication in the thin-flap LASIK. PMID- 19810663 TI - Intraocular straylight and contrast sensitivity after contralateral wavefront guided LASIK and wavefront-guided PRK for myopia. AB - PURPOSE: To compare intraocular straylight measurements and contrast sensitivity after wavefront-guided LASIK (WFG LASIK) in one eye and wavefront-guided photorefractive keratectomy (WFG PRK) in the fellow eye for myopia and myopic astigmatism correction. METHODS: A prospective, randomized study of 22 eyes of 11 patients who underwent simultaneous WFG LASIK and WFG PRK (contralateral eye). Both groups were treated with the NIDEK Advanced Vision Excimer Laser System, and a microkeratome was used for flap creation in the WFG LASIK group. High and low contrast visual acuity, wavefront analysis, contrast sensitivity, and retinal straylight measurements were performed preoperatively and at 3, 6, and 12 months postoperatively. A third-generation straylight meter, C-Quant (Oculus Optikgerate GmbH), was used for measuring intraocular straylight. RESULTS: Twelve months postoperatively, mean uncorrected distance visual acuity was -0.06 +/- 0.07 logMAR in the WFG LASIK group and -0.10 +/- 0.10 logMAR in the WFG PRK group. Mean preoperative intraocular straylight was 0.94 +/- 0.12 logs for the WFG LASIK group and 0.96 +/- 0.11 logs for the WFG PRK group. After 12 months, the mean straylight value was 1.01 +/- 0.1 log s for the WFG LASIK group and 0.97 +/- 0.12 log s for the WFG PRK group. No difference was found between techniques after 12 months (P = .306). No significant difference in photopic and mesopic contrast sensitivity between groups was noted. CONCLUSIONS: Intraocular straylight showed no statistically significant increase 1 year after WFG LASIK and WFG PRK. Higher order aberrations increased significantly after surgery for both groups. Nevertheless, WFG LASIK and WFG PRK yielded excellent visual acuity and contrast sensitivity performance without significant differences between techniques. PMID- 19810664 TI - The relationship between simulation in nursing education and medication safety. AB - This experimental study examined whether the use of clinical simulation in nursing education could help reduce medication errors. Fifty-four student volunteers were randomly assigned to an experimental (treatment) group (24 students) or a clinical control group (30 students). The treatment replaced some early-term clinical placement hours with a simulated clinical experience. The control group had all normally scheduled clinical hours. Treatment occurred prior to opportunities for medication administration. PMID- 19810665 TI - Incorporating community-based clinical experiences into a maternal-women's health nursing course. AB - Many barriers to increasing enrollment in nursing programs currently exist. One barrier is increased competition for clinical site placement from competing nursing and multidisciplinary programs. A critical component of nursing educational programs is providing clinical experiences that reinforce didactic learning. Incorporating women's health in a maternity course required clinical experiences to reflect this content. Faculty incorporated new strategies to help students integrate relevant theoretical concepts of maternity and women's health nursing by placing students in more community sites. Although labor and delivery and mother-baby units are located in the hospital, a wealth of community experiences for maternal and women's health nursing students exists. This article discusses strategies implemented by faculty at Youngstown State University to meet these challenges and promote appropriate clinical learning experiences for student nurses enrolled in a maternal-women's health nursing course. PMID- 19810666 TI - Using simulation to teach patient safety behaviors in undergraduate nursing education. AB - The purpose of this article is to describe how our college of nursing began to integrate patient safety instruction into simulation experiences for undergraduate nursing students. A system for evaluating and grading students was developed. Data on student safety behaviors were collected before and after implementation of instruction designed to improve adherence to hand washing and patient identification procedures. In the first semester in which data were collected, students did not demonstrate satisfactory performance of either hand hygiene or patient identification 61% of the time. After instruction, students still did not perform these procedures consistently 38% of the time. Lessons learned and future plans for addressing these problems with basic patient safety behaviors are discussed. PMID- 19810667 TI - Promoting empathy through a creative reflective teaching strategy: a mixed-method study. AB - The effects of a creative reflective clinical experience on psychiatric nursing students' empathy was examined using a mixed-method approach. The Interpersonal Reactivity Index (IRI) was used to measure empathy in students (N = 73) in a baccalaureate nursing program. The control group (n = 44) completed a conventional 14-week clinical experience, whereas the comparison group (n = 29) experienced a community psychosocial program for 4 of the weeks. Comparison group participants completed a creative reflective assignment to represent the life experience of a client with mental illness. To further understand empathy, a constructivist approach with purposive sampling was used to examine students' perceptions. Four themes emerged: having expectations, building relationships, changing perceptions, and gaining understanding through creative expression. Trends were noted, although no statistically significant differences in IRI scores were found for either group. However, this article argues that the use of a creative reflective assignment reduces stigma and promotes empathy. PMID- 19810668 TI - Equivalence testing of traditional and simulated clinical experiences: undergraduate nursing students' knowledge acquisition. AB - Although simulated clinical experience is being used increasingly in nursing education, vital evidence related to knowledge acquisition associated with simulated clinical experience does not exist. This intervention study used a 2*2 crossover design and equivalence testing to explore the effects of simulated clinical experiences on undergraduate students' (n = 74) knowledge acquisition in a fundamentals of nursing course. Following random assignment, students participated in laboratory-based simulated clinical experiences with high fidelity human patient simulators and traditional clinical experiences and completed knowledge pretests and posttests. Analysis identified significant knowledge gain associated with both simulated and traditional clinical experiences, with the groups' knowledge scores being statistically significantly equivalent. A priori equivalence bounds around the difference between the groups were set at +/- 5 points. Simulated clinical experience was found to be as effective as traditional clinical experience in promoting students' knowledge acquisition. PMID- 19810669 TI - Development and testing of a portfolio evaluation scoring tool. AB - This study focused on development of a portfolio evaluation tool to guide the assignment of valid and reliable scores. Tool development was facilitated by a literature review, guidance of a faculty committee, and validation by content experts. Testing involved a faculty team that evaluated 60 portfolios. Calculation of interrater reliability and a paired-samples t test were used to judge effectiveness. Interrater reliability was 0.78 for overall scores, 0.81 for the seven program outcomes criteria scores, and more than 0.65 for scores assigned by 11 of 13 pairs of raters. There were no significant differences between raters' scores in 10 of 13 pairs. The portfolio evaluation tool demonstrated high reliability and should be tested by other schools using portfolio evaluation. PMID- 19810670 TI - Innovative approach to teaching communication skills to nursing students. AB - This study assessed the effectiveness of a learner-centered simulation intervention designed to improve the communication skills of preprofessional sophomore nursing students. An innovative teaching strategy in which communication skills are taught to nursing students by using trained actors who served as standardized family members in a clinical learning laboratory setting was evaluated using a two-group posttest design. In addition to current standard education, the intervention group received a formal training session presenting a framework for communication and a 60-minute practice session with the standardized family members. Four domains of communication-introduction, gathering of information, imparting information, and clarifying goals and expectations-were evaluated in the control and intervention groups in individual testing sessions with a standardized family member. The intervention group performed better than the control group in all four tested domains related to communication skills, and the difference was statistically significant in the domain of gathering information (p = 0.0257). PMID- 19810671 TI - Benefits of using undergraduate teaching assistants throughout a baccalaureate nursing curriculum. AB - The Residency Model of Nursing Education was put into practice at our institution to provide more active teaching-learning strategies, make use of innovative clinical approaches, and accommodate more students. A unique aspect of this creative curricular change is the use of undergraduate teaching assistants (UTAs) to provide mentor and mentee opportunities for nursing students and assist faculty with course logistics. This article describes the development, challenges, and benefits of implementing a UTA program in a baccalaureate school of nursing. PMID- 19810672 TI - The development of evidence-based clinical simulation scenarios: guidelines for nurse educators. AB - Clinical simulation has been recognized as a teaching method using learning exercises that closely mimic real-life situations. The development of evidence based clinical simulation scenarios and guidelines for nurses is an important step in redesigning nursing education. These scenarios are created for students to learn in a safe environment. Simulated clinical experience requires immersing students in a representative patient-care scenario, a setting that mimics the actual environment with sufficient realism to allow learners to suspend disbelief. The purpose of this article is to discuss the Bay Area Simulation Collaborative's development of guidelines for effective evidence-based scenarios for use in hospitals and nursing schools. Six scholarly articles were reviewed and evaluated to determine whether evidence-based guidelines for scenario development exist and whether consensus in the literature regarding best practice is evident. PMID- 19810675 TI - Glucose-responsive micelles from self-assembly of poly(ethylene glycol)-b poly(acrylic acid-co-acrylamidophenylboronic acid) and the controlled release of insulin. AB - Poly(ethylene glycol)-block-poly(acrylic acid-co-acrylamidophenylboronic acid) [PEG(114)-b-(PAA(63)-co-PAAPBA(107))] was synthesized by the modification of poly(ethylene glycol)-block-poly(acrylic acid) (PEG(114)-b-PAA(170)) with 3 aminophenylboronic acid (APBA). Glucose-responsive PEG(114)-b-(PAA(63)-co PAAPBA(107)) self-assembled into core-shell micelles with the hydrophobic core composed of PAAPBA and hydrophilic shell composed of PEG in aqueous solution. The swelling and disaggregating behaviors of micelles responding to glucose were investigated by using light scattering in aqueous solution at pH 7.4. Characterization of insulin-loaded micelles and their drug release in solutions with various glucose concentrations were further studied. The results demonstrated that the drug release rate can be controlled by variation of glucose concentration. PMID- 19810673 TI - Regulating immune response using polyvalent nucleic acid-gold nanoparticle conjugates. AB - The immune response of macrophage cells to internalized polyvalent nucleic acid functionalized gold nanoparticles has been studied. This study finds that the innate immune response (as measured by interferon-beta levels) to densely functionalized, oligonucleotide-modified nanoparticles is significantly less (up to a 25-fold decrease) when compared to a lipoplex carrying the same DNA sequence. The magnitude of this effect is inversely proportional to oligonucleotide density. It is proposed that the enzymes involved in recognizing foreign nucleic acids and triggering the immune response are impeded due to the local surface environment of the particle, in particular high charge density. The net effect is an intracelluar gene regulation agent that elicits a significantly lower cellular immune response than conventional DNA transfection materials. PMID- 19810674 TI - Discovery of a novel class of potent coumarin monoamine oxidase B inhibitors: development and biopharmacological profiling of 7-[(3-chlorobenzyl)oxy]-4 [(methylamino)methyl]-2H-chromen-2-one methanesulfonate (NW-1772) as a highly potent, selective, reversible, and orally active monoamine oxidase B inhibitor. AB - In an effort to discover novel selective monoamine oxidase (MAO) B inhibitors with favorable physicochemical and pharmacokinetic profiles, 7-[(m halogeno)benzyloxy]coumarins bearing properly selected polar substituents at position 4 were designed, synthesized, and evaluated as MAO inhibitors. Several compounds with MAO-B inhibitory activity in the nanomolar range and excellent MAO B selectivity (selectivity index SI > 400) were identified. Structure-affinity relationships and docking simulations provided valuable insights into the enzyme inhibitor binding interactions at position 4, which has been poorly explored. Furthermore, computational and experimental studies led to the identification and biopharmacological characterization of 7-[(3-chlorobenzyl)oxy]-4 [(methylamino)methyl]-2H-chromen-2-one methanesulfonate 22b (NW-1772) as an in vitro and in vivo potent and selective MAO-B inhibitor, with rapid blood-brain barrier penetration, short-acting and reversible inhibitory activity, slight inhibition of selected cytochrome P450s, and low in vitro toxicity. On the basis of this preliminary preclinical profile, inhibitor 22b might be viewed as a promising clinical candidate for the treatment of neurodegenerative diseases. PMID- 19810676 TI - Solid-state (49/47)Ti NMR of titanium-based MCM-41 hybrid materials. AB - Titanium solid-state NMR spectroscopy data for a series of organic-inorganic titanium MCM-41 based materials have been collected. These materials have been synthesized by first modifying the mesoporous silica MCM-41 in one step with a mixture of silanes: a triazine propyl triethoxysilane acting as functional linker and methyltrimethoxysilane or hexamethyldisilizane as capped agents to mask the remaining silanol groups. Second, the appropiate titanium precursor Ti(OPr(i))(4), [{Ti(OPr(i))(3)(OMent)}(2)] (OMent = 1R,2S,5R-(-)-menthoxo), Ti(OPr(i))(4), or [Ti(eta(5)-C(5)HMe(4))Cl(3)], has been immobilized by reaction with the modified MCM-41. Finally, after Ti(OPr(i))(4) immobilization onto the organomodified support the reaction with the chiral (+)-diethyl-l-tartrate was accomplished. The materials without functional linker have been also prepared by reaction in one step of the capped agent and the titanium precursor with the mesoporous silica. Relevant correlations of titanium NMR resonance chemical shifts and line widths can be inferred depending on different factors. The immobilization procedure used to prepare titanium-based MCM-41 hybrid materials and the choice of the silylating reagents employed to mask the silanol groups present on the silica surfaces produce significant differences in the Ti NMR spectra. Furthermore, depending on the electronic and sterical influence of the substituents directly attached to the titanium center, chemical shifts and line widths are modified providing novel information about titanium structure. PMID- 19810678 TI - Imidazole-fe interaction in an aqueous chloride medium: effect of cathodic reduction of the native oxide. AB - The effect of the reduction of the native surface oxide of Fe on the binding of imidazole (as a corrosion inhibitor) with Fe in an aqueous brine solution has been addressed here. The surface interactions and corrosion inhibition efficiency were studied using X-ray photoelectron spectroscopy (XPS) and electrochemical impedance spectroscopy (EIS). It was shown that imidazole dissolved in brine bonds with the unreduced iron oxide surface via pyrrole-type nitrogen. However, surface interactions with Fe occur via both pyridine-type and pyrrole-type nitrogen atoms when imidazole is added to brine containing a cathodically reduced iron surface. The packing density of imidazole is found to be higher in the latter case with a corresponding increase in the corrosion inhibition efficiency. PMID- 19810677 TI - Simple large-scale synthesis of hydroxyapatite nanoparticles: in situ observation of crystallization process. AB - The noble synthesis method for hydroxyapatite (HAp) nanoparticles was exploited using a fairly simple reaction of Ca(OH)(2) and H(3)PO(4), which does not generate residual harmful anions and consequently does not need an additional washing process. HAp nanoparticles were found to yield from dicalcium phosphate dehydrate (DCPD) as the only intermediate phase, which was monitored by in situ observation study using X-ray diffraction (XRD), Fourier transform infrared (FT IR), (1)H and (31)P magic-angle spinning (MAS) NMR. Furthermore, we found that the phase evolution of HAp was preceded by heteronucleation of HAp onto the DCPD surface. The combination of scanning electron microscopy (SEM) and inductively coupled plasma atomic emission spectroscopy (ICP-ES) analysis gave more information on the HAp crystallization process, which was found to be retarded by the residual Ca(OH)(2) and slow diffusion process of Ca ions into the interface between HAp and DCPD. These results demonstrate that the synthesis of pure HAp nanoparticles with high throughput can be achieved by controlling the residual Ca(OH)(2) and diffusion process of Ca ions. PMID- 19810679 TI - Iron-induced cyclodextrin self-assembly into size-controllable nanospheres. AB - Iron-induced self-assembly of beta-cyclodextrin, beta-CD, into size controllable nanospheres with a well-defined spherical morphology and a relatively narrow size distribution was formed when acetone was added to a solution of beta-CD with iron(II) acetate, Fe(OAc)(2), in DMF. Thermogravimetric analysis, inductively coupled plasma atomic emission spectroscopy, and high resolution transmission electron microscopy showed that the irons were present as a well-dispersed state in the beta-CD nanospheres. In the (1)H NMR spectrum of the beta-CD/Fe(OAc)(2), beta/Fe, solution before adding acetone, the peaks corresponding to beta-CD were broadened and their spin-spin splitting had disappeared. In particular, the beta/Fe solutions were found to remain in a clean solution state at 1 week after solution preparation. These findings indicate the isolation of individual iron ions caused by the surrounding of each ion with the beta-CD molecules in the solution before the addition of acetone. X-ray crystal structure analysis, morphological observations, and N(2) adsorption and desorption experiments showed that the beta-CD nanospheres were generated by the formation of iron-embedded beta-CD primary particles with disordered cage type structure and simultaneous spherical assembly of the primary particles during the addition of acetone to the beta/Fe solution with appropriate mole ratio between beta-CD and Fe(OAc)(2). Interestingly, the size of the beta-CD nanospheres could be simply controlled by changing the speed at which acetone was added to the solution, with higher acetone addition speeds yielding smaller particles. PMID- 19810680 TI - Surface-mounted molecular rotors with variable functional groups and rotation radii. AB - A strategy for designing and activating surface-mounted molecular rotors with variable rotation radii and functional groups is proposed and demonstrated. The key point of the strategy is to separate the anchor and the rotating functional group from each other by using a connector of adjustable length. The three independent parts of the molecule are responsible for different functions to support the rotating movement of the molecule as a whole. In this way, one can easily change each part to obtain molecular rotors with different sizes, anchors, and functional rotating groups. PMID- 19810681 TI - New pathways in transannular cyclization of germacrone [germacra-1(10),4,7(11) trien-8-one]: evidence regarding a concerted mechanism. AB - The transannular cyclization of germacrone with HSO(3)Cl at -78 degrees C by means of a concerted and regioselective mechanism gives rise to a bicyclo[6.2.0]decan-2-ylium intermediate ion, which evolves to unusual skeletons through subsequent cyclization and Wagner-Meerwein rearrangements. This novel germacra-1(10),4-diene cyclization could suggest the existence of a new biosynthetic pathway to sesquiterpenes. PMID- 19810682 TI - Stereoselective synthesis of 2,6-syn-dimethyl-tetrahydropyran derivatives, important segments of marine polycyclic ethers, by unique insertion of the methyl group. AB - Treatment of 6-methyl-tetrahydropyran derivatives, which have a 1'-mesyloxy group at the C2-side chain, with Me(3)Al effected stereoselective insertion of a methyl group at the C2-position to give 2,6-syn-dimethyl-tetrahydropyran derivatives. This reaction proceeds via removal of the mesyloxy group, 1,2-hydride shift, and stereoselective insertion of a methyl group into the resulting oxonium ion. PMID- 19810683 TI - Nickel-catalyzed cross-coupling reaction of alkenyl methyl ethers with aryl boronic esters. AB - The Ni(0)-catalyzed cross-coupling of alkenyl methyl ethers with boronic esters is described. Several types of alkenyl methyl ethers can be coupled with a wide range of boronic esters to give the stilbene derivatives. PMID- 19810684 TI - Interplay between the transport of solutes across nanofiltration membranes and the thermal properties of the thin active layer. AB - The thin active layer (TAL) of seven nanofiltration (NF) membranes was studied using differential scanning calorimetry, and the membranes were classified into two groups according to the polymer physical state (amorphous or semicrystalline). NF membrane performance in terms of permeate volume flux density and rejection of neutral solutes was investigated in temperature cycles. The modeling of rejection using a hindered transport theory showed irreversible and opposite pore size changes for the two groups of NF membranes when the maximum operating temperature of the cycle exceeded the glass-transition temperature of the TAL. A mechanism of pore deformation is proposed to explain the variation of the solute transport properties as a function of the temperature and the polymer physical state in the TAL. PMID- 19810685 TI - Influence of surfactants on the force between two bubbles. AB - The retardation of the interfacial velocity due to the presence of surface-active species is a key feature that determines the magnitude of the dynamic interaction force between colliding bubbles. Here we derive simple measures to quantify the influence of a surface-active species during a head-on collision between bubbles to be used as guidelines in the design and analysis of emulsion stability and related experiments. These measures are derived from a theoretical model that was found to be consistent with experiment and are shown to characterize the interfacial dynamics without the need to use numerical analysis. It is shown that a surface mobility may change with the geometry of the film between the bubbles for a specific amount of a surface-active species. However, small amounts of surface-active species are sufficient to immobilize the interfaces under most physical conditions as found in earlier studies. PMID- 19810686 TI - Steady-state and time-resolved studies of the photocleavage of lysozyme by Co(III) complexes. AB - Steady-state and time-resolved studies of site-selective photocleavage of lysozyme by cobalt(III) complexes [Co(NH(3))(5)Br](2+) and ([Co(NH(3))(4)CO(3)](+) are reported. Photocleavage resulted in two fragments of molecular masses approximately 10.5 kDa and approximately 3.5 kDa, and the yield increased (8-33%) with irradiation time (0.16-0.8 h) as well as with the metal complex concentration (0.1-5 mM). The reaction proceeded to a significant extent even when nearly stoichiometric amounts of the reagents were used. Photocleavage was effective at wavelengths ranging from 310 to 390 nm, and cleavage was inhibited by the addition of selected metal ions such as Gd(III) at moderate concentrations (2 mM). Gd(III) is known to bind at Asp52/Glu35 residues on lysozyme, and these residues are located at the enzyme active site. Current and previous studies suggest that Co(III) metal complexes bind at this site on lysozyme. Consistent with this hypothesis, [Co(NH(3))(4)CO(3)](+) (8 mM) inhibited lysozyme activity by 67%. Laser flash photolysis studies show that excitation of the metal complexes [Co(NH(3))(5)Br](2+) and ([Co(NH(3))(4)CO(3)](+) (308 nm, 20 ns pulse width) resulted in the corresponding ligand-derived radical intermediates. For example, photoexcitation of an aqueous solution of [Co(NH(3))(5)Br](2+) at 308 nm resulted in the formation of Br(2)( *). When the excitation was carried out in the presence of lysozyme, Br(2)(-*) was quenched with a bimolecular rate constant of 1.4 x 10(9) M(-1) s(-1). Quenching resulted in protein-derived radicals (Trp(+*) and Tyr(+*)), as identified by their characteristic known transient absorption bands. Steady-state studies correlated with the time-resolved data, and taken together, these illustrated the reactivities of Co(III) metal complexes to direct protein photocleavage with high selectivity. PMID- 19810687 TI - Enantioselective enzymatic desymmetrization of highly functionalized meso tetrahydropyranyl diols. AB - The enantioselective enzymatic desymmetrization of several highly substituted meso-tetrahydropyranyl diols is described. This transformation leads to valuable building blocks containing up to five stereogenic centers, which are revealed in a single step with both high yields and excellent enantiomeric excesses. Moreover, it was shown that this kind of building blocks could provide an easy access to both enantiomers of highly functionalized stereotetrads. PMID- 19810688 TI - Substrate redox potential controls superoxide production kinetics in the cytochrome bc complex. AB - The Q-cycle mechanism of the cytochrome bc(1) complex maximizes energy conversion during the transport of electrons from ubiquinol to cytochrome c (or alternate physiological acceptors), yet important steps in the Q-cycle are still hotly debated, including bifurcated electron transport, the high yield and specificity of the Q-cycle despite possible short-circuits and bypass reactions, and the rarity of observable intermediates in the oxidation of quinol. Mounting evidence shows that some bypass reactions producing superoxide during oxidation of quinol at the Q(o) site diverge from the Q-cycle rather late in the bifurcated reaction and provide an additional means of studying initial reactions of the Q-cycle. Bypass reactions offer more scope for controlling and manipulating reaction conditions, e.g., redox potential, because they effectively isolate or decouple the Q-cycle initial reactions from later steps, preventing many complications and interactions. We examine the dependence of oxidation rate on substrate redox potential in the yeast cytochrome bc(1) complex and find that the rate limitation occurs at the level of direct one-electron oxidation of quinol to semiquinone by the Rieske protein. Oxidation of semiquinone and reduction of cyt b or O(2) are subsequent, distinct steps. These experimental results are incompatible with models in which the transfer of electrons to the Rieske protein is not a distinct step preceding transfer of electrons to cytochrome b, and with conformational gating models that produce superoxide by different rate-limiting reactions from the normal Q-cycle. PMID- 19810689 TI - Regioselective ruthenium-catalyzed direct benzylations of arenes through C-H bond cleavages. AB - Highly regioselective ruthenium-catalyzed direct benzylations through C-H bond cleavages were accomplished under remarkably mild, nonacidic reaction conditions, for which experimental studies suggested a S(E)Ar-type mechanism not to be operative. PMID- 19810690 TI - Apparent copper(II)-accelerated azide-alkyne cycloaddition. AB - Cu(II) salts accelerate azide-alkyne cycloaddition reactions in alcoholic solvents without reductants such as sodium ascorbate. Spectroscopic observations suggest that Cu(II) undergoes reduction to catalytic Cu(I) species via either alcohol oxidation or alkyne homocoupling, or both, during an induction period. The reactions involving 2-picolylazide are likely facilitated by its chelation to Cu(II). The highly exothermic reaction between 2-picolylazide and propargyl alcohol completes within 1-2 min in the presence of as low as 1 mol % Cu(OAc)(2). PMID- 19810691 TI - Synthesis of densely substituted pyrimidine derivatives. AB - The direct condensation of cyanic acid derivatives with N-vinyl/aryl amides affords the corresponding C4-heteroatom substituted pyrimidines. The use of cyanic bromide and thiocyanatomethane in this chemistry provides versatile azaheterocycles poised for further derivatization. The synthesis of a variety of previously inaccessible C2- and C4-pyrimidine derivatives using this methodology is described. PMID- 19810692 TI - Emission color tuning and deep blue dopant materials based on 1,6-bis(N-phenyl-p (R)-phenylamino)pyrene. AB - Panchromatic 1,6-bis(N-phenyl-p-(R)-phenylamino)pyrenes, 2R, were obtained from Buchwald-Hartwig coupling reactions between N-phenyl-p-(R)-phenylamines and 1,6 dibromopyrene. The photophysical properties of 2R corresponded well to the electron-withdrawing and -donating nature of the diarylamine substituents, exhibiting a full color visible range between 454 and 620 nm. In particular, a deep blue 2CN showed a high radiative rate constant of 2.85 x 10(8) s(-1) with high emission quantum efficiency of 79%. Further applications of 2CN as a blue dopant were attempted using multilayer organic light-emitting devices. A maximum efficiency of 3.98 cd/A with CIE coordinates of x = 0.14, y = 0.10 were obtained. PMID- 19810693 TI - Synthesis of a cyclic pentapeptide mimic of the active site His-Tyr cofactor of cytochrome c oxidase. AB - Arylboronic acid based technology provides a mild, regioselective, and nontoxic N arylation procedure for accessing the unusual N-arylated side chain histidine found in the active site of cytochrome c oxidase (CcO). The N-arylated histidine is elaborated to the complete cytochrome c oxidase cyclic pentapeptide cofactor. Molecular modeling of the cofactor provides insight into the dynamic character of the N-aryl bond. PMID- 19810695 TI - Strong specific hydroxide ion binding at the pristine oil/water and air/water interfaces. AB - Despite claims, based largely on molecular dynamics simulations, that the surface of water at the air/water interface is acidic, with a positive charge, there is compelling experimental evidence that it is in fact basic, with a negative charge due to the specific adsorption of hydroxide ions. The oil/water interface behaves similarly. The pH dependence of the zeta potentials of oil drops has been measured by two very different techniques: on a single drop in a rotating electrophoresis cell and on about 10(14) submicrometer drops in a 2 vol % emulsion by an electroacoustic method to give similar results with a sigmoidal pH dependence characterized by an isoelectric point at pH 2-3 and a half adsorption point about pH 5.5, or at 10(-8.5) M hydroxide ion. This indicates that hydroxide ion is absorbed much more strongly than other anions. The pH dependence of a single N(2) bubble has also been measured and has the same pH dependence, independently of whether HCl or HI is used to adjust the pH. These similarities between the pH dependences of the zeta potentials of air bubbles and oil drops, as well as those reported from streaming potentials on solid inert surfaces such as Teflon, indicate that water behaves similarly, with only subtle differences, at each of these low dielectric hydrophobic surfaces, with an isoelectric point of pH 2-4. In acidic solutions at pH's below the isoelectric point, the surface is indeed positive, consistent with spectroscopic observations of the adsorption of hydrogen ions. PMID- 19810694 TI - Comparing the properties of electrochemical-based DNA sensors employing different redox tags. AB - Many electrochemical biosensor approaches developed in recent years utilize redox labeled (most commonly methylene blue or ferrocene) oligonucleotide probes site specifically attached to an interrogating electrode. Sensors in this class have been reported that employ a range of probe architectures, including single- and double-stranded DNA, more complex DNA structures, DNA and RNA aptamers, and, most recently, DNA-small molecule chimeras. Signaling in this class of sensors is generally predicated on binding-induced changes in the efficiency with which the covalently attached redox label transfers electrons with the interrogating electrode. Here we have investigated how the properties of the redox tag affect the performance of such sensors. Specifically, we compare the differences in signaling and stability of electrochemical DNA sensors (E-DNA sensors) fabricated using either ferrocene or methylene blue as the signaling redox moiety. We find that while both tags support efficient E-DNA signaling, ferrocene produces slightly improved signal gain and target affinity. These small advantages, however, come at a potentially significant price: the ferrocene-based sensors are far less stable than their methylene blue counterparts, particularly with regards to stability to long-term storage, repeated electrochemical interrogations, repeated sensing/regeneration iterations, and employment in complex sample matrices such as blood serum. PMID- 19810696 TI - Fluorescence-based detection of point mutation in DNA sequences by CdS quantum dot aggregation. AB - We present a novel method for the detection of single base mismatch based on fluorescence quenching that unmodified CdS quantum dots exhibit upon aggregation. Target DNA sequences of interest are breast cancer 2 (BRCA2) and signal-induced proliferation-associated gene 1 (Sipa1) sequences. We monitor aggregation of CdS quantum dots upon addition of double-stranded DNAs at different salt concentration using quasi-elastic light scattering (QELS), transmission electron microscopy (TEM), photoluminescence spectroscopy, and zeta potential measurement. Our results indicate that the double-stranded DNA with a perfectly matched sequence can easily be discerned by naked eye from the single base mismatched one due to the fluorescence quenching phenomenon caused by selective aggregation of the CdS quantum dots. PMID- 19810697 TI - Expedited solid-phase synthesis of fluorescently labeled and biotinylated aminoalkane diphenyl phosphonate affinity probes for chymotrypsin- and elastase like serine proteases. AB - In this study, we report on a novel, expedited solid-phase approach for the synthesis of biotinylated and fluorescently tagged irreversible affinity based probes for the chymotrypsin and elastase-like serine proteases. The novel solid phase biotinylation or fluorescent labeling of the aminoalkane diphenyl phosphonate warhead using commercially available Biotin-PEG-NovaTag or EDANS NovaTag resin permits rapid, facile synthesis of these reagents. We demonstrate the kinetic evaluation and utilization of a number of these irreversible inactivators for chymotrypsin-like (chymotrypsin/human cathepsin G) and elastase like serine proteases. Encouragingly, these compounds display comparable potency against their target proteases as their N-benzyloxycarbonyl (Cbz)-protected parent compounds, from which they were derived, and function as efficient active site-directed inactivators of their target proteases. We subsequently applied the biotinylated reagents for the sensitive detection of protease species via Western blot, showing that the inactivation of the protease was specifically mediated through the active site serine. Furthermore, we also demonstrate the successful detection of serine protease species with the fluorescently labeled derivatives "in-gel", thus avoiding the need for downstream Western blotting. Finally, we also show the utility of biotinylated and pegylated affinity probes for the isolation/enrichment of serine protease species, via capture with immobilized streptavidin, and their subsequent identification via de novo sequencing. Given their selectivity of action against the serine proteases, we believe that these reagents can be exploited for the direct, rapid, and selective identification of these enzymes from biological milieu containing multiple protease subclasses. PMID- 19810698 TI - Analysis of the dynamics of assembly and structural impact for a histidine tagged FGF1-1.5 nm Au nanoparticle bioconjugate. AB - Whether assembling proteins onto nanoscale, mesoscopic, or macroscropic material surfaces, maintaining a protein's structure and function when conjugated to a surface is complicated by the high propensity for electrostatic or hydrophobic surface interactions and the possibility of direct metal coordination of protein functional groups. In this study, the assembly of a 1.5 nm CAAKA passivated gold nanoparticle (AuNP) onto FGF1 (human acidic fibroblast growth factor) using an amino terminal His(6) tag is analyzed. The impact of structure and time-dependent changes in the structural elements in FGF1and FGF1-heparin in the presence of the AuNP is probed by a molecular beacon fluorescence assay, circular dichroism, and NMR spectroscopy. Analysis of the results indicates that a time-dependent evolution of the protein structure without loss of FGF1 heparin binding occurs following the formation of the initial FGF1-AuNP complex. The time-dependent changes are believed to reflect protein sampling of the AuNP surface to minimize the free energy of the AuNP-FGF1 complex without impacting FGF1 function. PMID- 19810699 TI - Thermodynamics, optical properties, and coordination modes of Np(V) with dipicolinic acid. AB - Complexation of NpO(2)(+) with dipicolinic acid (DPA) has been investigated in 1 M NaClO(4) at 25 degrees C. Two complexes, NpO(2)(DPA)(-) and NpO(2)(DPA)(2)(3-), were identified, and the stability constants (log beta(1) and log beta(2)) were determined to be 8.68 and 12.31, respectively, by spectrophotometry. The enthalpies of the complexation (DeltaH(1) and DeltaH(2)) were measured to be 25.2 and -45.9 kJ/mol by microcalorimetry. The entropies (DeltaS(1) and DeltaS(2)) were calculated to be 81.6 and 81.8 J/(K mol) accordingly. The strong complexation of NpO(2)(+) with DPA is driven by both positive entropies and highly exothermic enthalpies. The crystal structure of Na(3)NpO(2)(DPA)(2)(H(2)O)(6)(s) shows that, in the NpO(2)(DPA)(2)(3-) complex, the Np atom sits at a center of inversion and the two DPA ligands symmetrically coordinate to Np in a tridentate mode. Due to the centrosymmetric structure of the NpO(2)(DPA)(2)(3-) complex, the f --> f transitions of Np(V) are forbidden, and the sharp bands originating from the f --> f transitions either disappear or become very weak in the optical absorption or diffuse reflectance spectra of the NpO(2)(DPA)(2)(3-) complex. PMID- 19810700 TI - Chemical interaction in the invasiveness of cogongrass (Imperata cylindrica (L.) Beauv.). AB - From gas chromatography-mass spectrometry (GC-MS), numerous plant growth inhibitors were found in the rhizome and root exudates of cogongrass, one of the most problematic weeds in the world. iso-Eugenol, iso-ferulic acid, linoleic acid, ferulic acid, and vanillin were the major chemicals in the rhizome (88.1 392.2 microg/g of fresh root), while 4-acetyl-2-methoxyphenol was the principle substance (872.6 microg/plant) in the root exudates. In fields, the use of cutting and plowing reduced weed biomass and weed density of cogongrass >70%. However, the alternative invasion of beggar tick might be a problem, because its density and biomass increased 33.3 and 62.5%, respectively. Chemicals from cogongrass showed selective effects against tested invasive species. Of them, 2,4 di-tert-butylphenol was the most potent (78.3-100% of inhibition), followed by iso-eugenol and 4-acetyl-2-methoxyphenol. These compounds may play important roles in the invasiveness of cogongrass and might be promising parent constituents of synthesis to develop novel herbicides for control of invasive plants. PMID- 19810701 TI - Meso-unsubstituted iron corrole in hemoproteins: remarkable differences in effects on peroxidase activities between myoglobin and horseradish peroxidase. AB - Myoglobin (Mb) and horseradish peroxidase (HRP) were both reconstituted with a meso-unsubstituted iron corrole and their electronic configurations and peroxidase activities were investigated. The appearance of the 540 nm band upon incorporation of the iron corrole into apoMb indicates axial coordination by the proximal histidine imidazole in the Mb heme pocket. Based on (1)H NMR measurements using the Evans method, the total magnetic susceptibility of the iron corrole reconstituted Mb was evaluated to be S = 3/2. In contrast, although a band does not appear in the vicinity of 540 nm during reconstitution of the iron corrole into the matrix of HRP, a spectrum similar to that of the iron corrole reconstituted Mb is observed upon the addition of dithionite. This observation suggests that the oxidation state of the corrole iron in the reconstituted HRP can be assigned as +4. The catalytic activities of both proteins toward guaiacol oxidation are quite different; the iron corrole reconstituted HRP decelerates H(2)O(2)-dependent oxidation of guaiacol, while the same reaction catalyzed by iron corrole reconstituted Mb has the opposite effect and accelerates the reaction. This finding can be attributed to the difference in the oxidation states of the corrole iron when these proteins are in the resting state. PMID- 19810702 TI - Understanding photocatalytic metallization of preadsorbed ionic gold on titania, ceria, and zirconia. AB - A nonaqueous photodeposition procedure for forming Au nanoparticles on semiconducting supports (TiO(2), CeO(2), and ZrO(2)) was investigated. Intrinsic excitation of the support was sufficient to induce Au(0) nucleation, without the need for an organic hole-scavenging species. Photoreduction rates were higher over TiO(2) and ZrO(2) than over CeO(2), likely due to a lower rate of photogenerated electron recombination. Illumination resulted in metallization of the adsorbed Au species and formation of crystalline Au nanoparticles dispersed across the oxide surfaces. On the basis of transmission electron microscopy (TEM) evidence of a strong Au particle-metal oxide interaction, it is proposed that Au deposit formation proceeds via the nucleation of highly dispersed clusters which can diffuse and amalgamate at room temperature to form larger surface-defect immobilized clusters, with the final particle size being significantly smaller than that achieved by conventional aqueous photodeposition. From this work, it is possible to draw several new fundamental insights, with regards to both the nonaqueous photodeposition process and the general mechanism by which dispersed metallic Au nanoparticles are formed from ionic precursors adsorbed upon metal oxide supports. PMID- 19810703 TI - Knowledge-based characterization of similarity relationships in the human protein tyrosine phosphatase family for rational inhibitor design. AB - Tyrosine phosphorylation, controlled by the coordinated action of protein tyrosine kinases (PTKs) and protein-tyrosine phosphatases (PTPs), is a fundamental regulatory mechanism of numerous physiological processes. PTPs are implicated in a number of human diseases, and their potential as prospective drug targets is increasingly being recognized. Despite their biological importance, until now no comprehensive overview has been reported describing how all members of the human PTP family are related. Here we review the entire human PTP family and present a systematic knowledge-based characterization of global and local similarity relationships, which are relevant for the development of small molecule inhibitors. We use parallel homology modeling to expand the current PTP structure space and analyze the human PTPs based on local three-dimensional catalytic sites and domain sequences. Furthermore, we demonstrate the importance of binding site similarities in understanding cross-reactivity and inhibitor selectivity in the design of small molecule inhibitors. PMID- 19810704 TI - Metabolic effects of dark chocolate consumption on energy, gut microbiota, and stress-related metabolism in free-living subjects. AB - Dietary preferences influence basal human metabolism and gut microbiome activity that in turn may have long-term health consequences. The present study reports the metabolic responses of free living subjects to a daily consumption of 40 g of dark chocolate for up to 14 days. A clinical trial was performed on a population of 30 human subjects, who were classified in low and high anxiety traits using validated psychological questionnaires. Biological fluids (urine and blood plasma) were collected during 3 test days at the beginning, midtime and at the end of a 2 week study. NMR and MS-based metabonomics were employed to study global changes in metabolism due to the chocolate consumption. Human subjects with higher anxiety trait showed a distinct metabolic profile indicative of a different energy homeostasis (lactate, citrate, succinate, trans-aconitate, urea, proline), hormonal metabolism (adrenaline, DOPA, 3-methoxy-tyrosine) and gut microbial activity (methylamines, p-cresol sulfate, hippurate). Dark chocolate reduced the urinary excretion of the stress hormone cortisol and catecholamines and partially normalized stress-related differences in energy metabolism (glycine, citrate, trans-aconitate, proline, beta-alanine) and gut microbial activities (hippurate and p-cresol sulfate). The study provides strong evidence that a daily consumption of 40 g of dark chocolate during a period of 2 weeks is sufficient to modify the metabolism of free living and healthy human subjects, as per variation of both host and gut microbial metabolism. PMID- 19810705 TI - Surface modification of PLGA particles: the interplay between stabilizer, ligand size, and hydrophobic interactions. AB - Therapeutic and diagnostic carriers can be functionalized with active targeters to induce tissue-specific delivery. However, the possible impact of adsorbed steric stabilizer such as the frequently used poloxamers (Pluronics) on surface modification of poly(D,L-lactide-co-glycolide) (PLGA) particles has not been examined so far. Therefore, three model ligands of different molecular weights (653; 36,000; 155,000 g/mol) covering the size range of important targeters were conjugated to the surface of PLGA microparticles in the presence of different concentrations of Pluronic F68 (0.01-5%, w/v). Flow cytometry and fluorimetric quantification revealed for all tested ligands that high Pluronic concentrations decreased the coupling efficiency to a half or even one-third of that achieved in the absence of stabilizer. Moreover, the reduction strongly depends on the ligand size and its propensity for hydrophobic interactions. Apart from that, a high degree of particle aggregation was observed with Pluronic concentrations below 0.1% (w/v). Thus, a compromise has to be found, which combines sufficient stability with the best possible ligand coupling efficiency. For the studied system, 0.1% (w/v) turned out to be the optimum concentration of Pluronic F68. PMID- 19810706 TI - The SufBCD Fe-S scaffold complex interacts with SufA for Fe-S cluster transfer. AB - Iron-sulfur clusters are key iron cofactors in biological pathways ranging from nitrogen fixation to respiration. Because of the toxicity of ferrous iron and sulfide to the cell, in vivo Fe-S cluster assembly transpires via multiprotein biosynthetic pathways. Fe-S cluster assembly proteins traffic iron and sulfide, assemble nascent Fe-S clusters, and correctly transfer Fe-S clusters to the appropriate target metalloproteins in vivo. The Gram-negative bacterium Escherichia coli contains a stress-responsive Fe-S cluster assembly system, the SufABCDSE pathway, that functions under iron starvation and oxidative stress conditions that compromise Fe-S homeostasis. Using a combination of protein protein interaction and in vitro Fe-S cluster assembly assays, we have characterized the relative roles of the SufBCD complex and the SufA protein during Suf Fe-S cluster biosynthesis. These studies reveal that SufA interacts with SufBCD to accept Fe-S clusters formed de novo on the SufBCD complex. Our results represent the first biochemical evidence that the SufBCD complex within the Suf pathway functions as a novel Fe-S scaffold system to assemble nascent clusters and transfer them to the SufA Fe-S shuttle. PMID- 19810707 TI - Coverage dependence of the adsorption structure of alanine on Ge(100). AB - The variations with coverage and annealing temperature in the adsorption structure of alanine on Ge(100) have been investigated using high-resolution core level photoemission spectroscopy (HRCLPES). The C 1s, N 1s, and O 1s core-level spectra at a low initial coverage show that both the carboxyl and amine groups of the alanine molecules participate in bonding with the Ge(100) surface in an "intrarow O-H dissociated and N dative bonded structure". However, at higher coverage we found that in addition to this structure an "O-H dissociation structure" is present. Moreover, we systematically monitored the variation of the bonding features of alanine adsorbed on Ge(100) with annealing temperature and thus were able to track the desorption processes. By analyzing the C 1s, N 1s, and O 1s spectra at 420 K, we conclude that the principal adsorption structure at this temperature is the "O-H dissociation structure" because of the disconnection of Ge-N dative bonding. At higher temperatures, the "O-H dissociation structure" is converted into various fragments such as Ge oxide (or Ge-CO), Ge nitride (Ge cyanide), and Ge carbide. PMID- 19810708 TI - High density labeling of polymerase chain reaction products with the fluorescent base analogue tCo. AB - Fluorescent DNA of high molecular weight is an important tool for studying the physical properties of DNA and DNA-protein interactions, and it plays a key role in modern biotechnology for DNA sequencing and detection. While several DNA polymerases can incorporate large numbers of dye-linked nucleotides into primed DNA templates, the amplification of the resulting densely labeled DNA strands by polymerase chain reaction (PCR) is problematic. Here, we report a method for high density labeling of DNA in PCR reactions employing the 5'-triphosphate of 1,3 diaza-2-oxo-phenoxazine (tCo) and Deep Vent DNA polymerase. tCo is a fluorescent cytosine analogue that absorbs and emits light at 365 and 460 nm, respectively. We obtained PCR products that were fluorescent enough to directly visualize them in a gel by excitation with long UV light, thus eliminating the need for staining with ethidium bromide. Reactions with Taq polymerase failed to produce PCR products in the presence of only small amounts of dtCoTP. A comparative kinetic study of Taq and Deep Vent polymerase revealed that Taq polymerase, although it inserts dtCoTP with high efficiency opposite G, is prone to forming mutagenic tCo A base pairs and does not efficiently extend base pairs containing tCo. These kinetics features explain the poor outcome of the PCR reactions with Taq polymerase. Since tCo substitutes structurally for cytosine, the presented labeling method is believed to be less invasive than labeling with dye-linked nucleotides and, therefore, produces DNA that is ideally suited for biophysical studies. PMID- 19810709 TI - Preparation and optimization of iridium complexes with quinazolinone ligands on solid supports. PMID- 19810711 TI - Mathematical approach by a selectivity model for rationalization of pH- and selector concentration-dependent reversal of the enantiomer migration order in capillary electrophoresis. AB - A separation selectivity model for capillary electrophoresis enantioseparations of weak bases in the presence of uncharged chiral selectors was described as a function of buffer pH and chiral selector concentration. On the basis of the selectivity at the extreme pH and selector concentration values, 15 principal cases could be distinguished describing the migration behavior of the analytes when increasing either pH or selector concentration. A pH-dependent reversal of the enantiomer migration order can be observed (1) when the complex mobility ratio is reversed due to an enantioselective complexation-induced pK(a)-shift, (2) in the case of an inversion of the ratio of the mobilities of the analyte selector complex and the free analyte due to significantly weaker complexation of the neutral species, and (3) in the case of opposite chiral recognition of the protonated and uncharged species by the chiral selector. Reversal of enantiomer migration order as a function of the selector concentration at a fixed buffer pH is caused by the opposing effects of complexation constants and complex mobilities at the respective pH value. The model was applied to rationalize the enantioseparations of dipeptides, especially the pH-dependent reversal of the migration order of the LL- and DD-enantiomers of Ala-Tyr in the presence of heptakis-(2,6-di-O-methyl)-beta-cyclodextrin as the chiral selector based on the complexation constants of the protonated and zwitterionic forms of the analytes. PMID- 19810712 TI - Diffraction patterns of a water-submerged superhydrophobic grating under pressure. AB - We report on a study of superhydrophobic surfaces submerged in water in a fluidic chamber. A surface-treated transmission grating was used as a superhydrophobic layer that had a well-defined diffraction pattern when a laser beam passed through the water-submerged grating sample, indicating a Cassie-Baxter state with trapped air between the water and grating interfaces. By applying pressure to the water in the fluidic chamber, the diffraction pattern can be changed because of the volume reduction of trapped air or water penetration into the grating. Depending on the maximum value of applied pressure in the fluidic chamber, the diffraction pattern change can be either reversible or irreversible after the release of the pressure. We attribute the irreversible change under high applied pressure to the switching from a Cassie-Baxter state to a Wenzel state. PMID- 19810710 TI - Lipid profiles of canine invasive transitional cell carcinoma of the urinary bladder and adjacent normal tissue by desorption electrospray ionization imaging mass spectrometry. AB - Desorption electrospray ionization mass spectrometry (DESI-MS) was used in an imaging mode to interrogate the lipid profiles of thin tissue sections of canine spontaneous invasive transitional cell carcinoma of the urinary bladder (a model of human invasive bladder cancer) as well as adjacent normal tissue from four different dogs. The glycerophospholipids and sphingolipids that appear as intense signals in both the negative ion and positive ion modes were identified by tandem mass spectrometry product ion scans using collision-induced dissociation. Differences in the relative distributions of the lipid species were present between the tumor and adjacent normal tissue in both the negative and positive ion modes. DESI-MS images showing the spatial distributions of particular glycerophospholipids, sphinoglipids, and free fatty acids in both the negative and positive ion modes were compared to serial tissue sections that were stained with hematoxylin and eosin (H&E). Increased absolute and relative intensities for at least five different glycerophospholipids and three free fatty acids in the negative ion mode and at least four different lipid species in the positive ion mode were seen in the tumor region of the samples in all four dogs. In addition, one sphingolipid species exhibited increased signal intensity in the positive ion mode in normal tissue relative to the diseased tissue. Principal component analysis was also used to generate unsupervised statistical images from the negative ion mode data, and these images are in excellent agreement with the DESI images obtained from the selected ions and also the H&E-stained tissue. PMID- 19810713 TI - Bifunctionalized hybrid silica spheres by hydrolytic cocondensation of 3 aminopropyltriethoxysilane and vinyltriethoxysilane. AB - Facile, surfactant free synthetic strategy for bifunctionalized hybrid silica spheres (HS) with structural ordering is presented. HS was prepared by casting and drying of stable siloxane solution from hydrolytic co-condensation of 3 aminopropyltriethoxysilane (AS) and vinyltriethoxysilane (VS) with AS:VS mole ratio of 1:3 in ethanol/water mixture. Spheres of size in the range of 250 nm to 2.5 microm were produced by adjusting the concentration of reacted siloxane solution using ethanol. Characterization by FTIR, XRD, TGA, and DSC revealed that the HS was formed from coprecipitation of fully condensed polyhedral oligomeric silsesquioxane (POSS) bilayer and incompletely condensed siloxanes (SIL) produced during drying. Formation of POSS bilayer was confirmed by intercalating and stabilizing POSS within a smectite clay and characterizing the modified clay. XRD, FTIR, SEM, and HRTEM of HS heated to 170 degrees C revealed transformation from disordered into ordered lamellar structure of POSS bilayer assembly and siloxane network due to rearrangement and densification of low melting SIL. PMID- 19810714 TI - From classical adducts to frustrated Lewis pairs: steric effects in the interactions of pyridines and B(C6F5)3. AB - The pyridine adducts of B(C(6)F(5))(3), (4-tBu)C(5)H(4)NB(C(6)F(5))(3) 1, ((2 Me)C(5)H(4)N)B(C(6)F(5))(3) 2, ((2-Et)C(5)H(4)N)B(C(6)F(5))(3) 3, ((2 Ph)C(5)H(4)N)B(C(6)F(5))(3) 4, ((2-C(5)H(4)N)C(5)H(4)N)B(C(6)F(5))(3) 5, (C(9)H(7)N)B(C(6)F(5))(3) 6, and ((2-C(5)H(4)N)NH(2-C(5)H(4)N))B(C(6)F(5))(3) 7, were prepared and characterized. The B-N bond lengths in 2-7 reflect the impact of ortho-substitution, increasing significantly with sterically larger and electron-withdrawing substituents. In the case of 2-amino-6-picoline, reaction with B(C(6)F(5))(3) affords the zwitterionic species (5-Me)C(5)H(3)NH(2 NH)B(C(6)F(5))(3) 8. In contrast, lutidine/B(C(6)F(5))(3) yields an equilibrium mixture containing both the free Lewis acid and base and the adduct (2,6 Me(2)C(5)H(3)N)B(C(6)F(5))(3) 9. This equilibrium has a DeltaH of -42(1) kJ/mol and DeltaS of -131(5) J/mol x K. Addition of H(2) shifts the equilibrium and yields [2,6-Me(2)C(5)H(3)NH][HB(C(6)F(5))(3)] 10. The corresponding reactions of 2,6-diphenylpyridine or 2-tert-butylpyridine with B(C(6)F(5))(3) showed no evidence of adduct formation and upon exposure to H(2) afforded [(2,6 Ph(2))C(5)H(3)NH][HB(C(6)F(5))(3)] 11 and [(2-tBu)C(5)H(4)NH][HB(C(6)F(5))(3)] 12, respectively. The energetics of adduct formation and the reactions with H(2) are probed computationally. Crystallographic data for compounds 1-10 are reported. PMID- 19810715 TI - Phase transition in salt-free catanionic surfactant mixtures induced by temperature. AB - Aggregate transitions in salt-free catanionic surfactant mixtures of tetradecyltrimethylammonium hydroxide (TTAOH)/fatty acid were investigated as a function of surfactant concentration and temperature. Lauric acid (LA), myristic acid (MA), and palmitic acid (PA) were chosen for the current study. The TTAOH/LA mixture exhibited rich phase behavior at room temperature. With increasing total surfactant concentration (c(T)), a bluish vesicular (L(alphav)) phase, an isotropic micellar (L(1)) phase, and a birefringent lamellar (L(alpha)) phase were observed. Between the L(alphav) phase and the L(1) phase, a narrow L(alpha)'/L(1) two-phase region was determined. With increasing temperature, a transition from the L(alpha) phase to the L(1) phase was induced at higher c(T) whereas at lower c(T) an opposite transition from the L(1) phase to the L(alphav) phase was noticed. Thus surprisingly, we observed bilayer-to-micelle and micelle to-bilayer transitions in the same catanionic surfactant system, both induced by the temperature increase. Replacing LA by MA and PA caused a continuous increase in the average Krafft point of the mixture. The L(alphav)-phase region and phase separated region become larger. Moreover, a single L(1)-phase region was absent within the investigated temperature range. PMID- 19810716 TI - Deterministic lateral displacement as a means to enrich large cells for tissue engineering. AB - The enrichment or isolation of selected cell types from heterogeneous suspensions is required in the area of tissue engineering. State of the art techniques utilized for this separation include preplating and sieve-based approaches that have limited ranges of purity and variable yield. Here, we present a deterministic lateral displacement (DLD) microfluidic device that is capable of separating large epithelial cells (17.3 +/- 2.7 in diameter) from smaller fibroblast cells (13.7 +/- 3.0 microm in diameter) as a potential alternative approach. The mixed suspension examined is intended to represent the content of digested rat cardiac tissue, which contains equal proportions of cardiomyocyte (17.0 +/- 4.0 microm diameter) and nonmyocyte populations (12.0 +/- 3.0 microm diameter). High purity separation (>97%) of the larger cell type is achieved with 90% yield in a rapid and single-pass process. The significance of this work lies in the recognition that DLD design principles can be applied for the microfluidic enrichment of large cells, up to the 40 microm diameter level examined in this work. PMID- 19810717 TI - Improved enzyme immobilization on an ionic-complementary peptide-modified electrode for biomolecular sensing. AB - An ionic-complementary peptide, EFK16-II, was used to modify a highly ordered pyrolytic graphite (HOPG) electrode surface. The nanofiber structure of the self assembling peptide on the electrode surface was characterized by atomic force microscopy (AFM). Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) spectra showed that, upon addition of 1-ethyl-3-(3 dimethylaminopropyl) carbodiimide (EDC), EFK16-II molecules tend to be cross linked among themselves. Cross-linking of the peptide diminishes the number of carboxyl groups available for immobilizing a sensing enzyme, which is glucose oxidase (GOx) in the present case. A simple method based on premixing of the carbodiimide and GOx was developed; it inhibited peptide cross-linking and significantly improved enzyme immobilization. Biosensors constructed in this way showed increased overall signal intensity and a much higher sensitivity at 4.94 mA M(-1) cm(-2), a 6-fold increase compared to the previously reported peptide modified electrodes. The results presented here demonstrate significant potential for ionic-complementary peptides for constructing electrochemical biosensors. PMID- 19810718 TI - Site-selective biofunctionalization of aluminum nitride surfaces using patterned organosilane self-assembled monolayers. AB - Surface biochemical functionalization of group-III nitride semiconductors has recently attracted much interest because of their biocompatibility, nontoxicity, and long-term chemical stability under demanding physiochemical conditions for chemical and biological sensing. Among III-nitrides, aluminum nitride (AlN) and aluminum gallium nitride (AlGaN) are particularly important because they are often used as the sensing surfaces for sensors based on field-effect transistor or surface acoustic wave (SAW) sensor structures. To demonstrate the possibility of site-selective biofunctionalization on AlN surfaces, we have fabricated two dimensional antibody micropatterns on AlN surfaces by using patterned self assembled monolayer (SAM) templates. Patterned SAM templates are composed of two types of organosilane molecules terminated with different functional groups (amino and methyl), which were fabricated on AlN/sapphire substrates by combining photolithography, lift-off process, and self-assembly technique. Because the patterned SAM templates have different surface properties on the same surface, clear imaging contrast of SAM micropatterns can be observed by field-emission scanning electron microscopy (FE-SEM) operating at a low accelerating voltage in the range of 0.5-1.5 kV. Furthermore, the contrast in surface potential of the binary SAM microstructures was confirmed by selective adsorption of negatively charged colloidal gold nanoparticles (AuNPs). The immobilization of AuNPs was limited on the positively charged amino-terminated regions, while they were scarcely found on the surface regions terminated by methyl groups. In this work, selective immobilization of green fluorescent protein (GFP) antibodies was demonstrated by the specific protein binding of enhanced GFP (EGFP) labeling. The observed strong fluorescent signal from antibody functionalized regions on the SAM-patterned AlN surface indicates the retained biological activity of specific molecular recognition resulting from the antibody-EGFP interaction. The results reported here show that micropatterning of organosilane SAMs by the combination of photolithographic process and lift-off technique is a practical approach for the fabrication of reaction regions on AlN-based bioanalytical microdevices. PMID- 19810719 TI - A photocontrolled molecular switch regulates paralysis in a living organism. AB - Using light to modulate biochemical agents in living organisms has a significant impact on photodynamic therapy and drug release. We demonstrate that a photoresponsive system can reversibly induce paralysis in nematodes as a model for living organisms when two different wavelengths of light are used to toggle the molecular switch between its two structural forms. This example illustrates how photoswitches offer great potential for advancing biomedical technologies. PMID- 19810720 TI - UV-defined flat PDMS stamps suitable for microcontact printing. AB - We report a simple method of creating well-defined micropatterns on the surface of a flat PDMS stamp, making it suitable for microcontact printing of proteins. This method only requires a UV lamp (254 nm) and a TEM grid (as a photomask) to modify the surface of PDMS for creating desired micropatterns. By using the UV modified stamp, a printed protein micropattern that resembles the original TEM grid can be obtained. Surprisingly, unlike the oxygen-plasma-treated PDMS, the UV modified flat stamp is also long-lasting (>1 week). The method reported herein is very economical for microcontact printing applications because expensive silicon masters and microstructured PDMS are no longer required. PMID- 19810721 TI - Quantitative photochemical formation of [Ru(tpy)(bpy)H]+. AB - Quantitative photochemical production of [Ru(tpy)(bpy)H](+) (Ru-H(+)) was achieved by irradiation of [Ru(tpy)(bpy)(DMF)](2+) (Ru-DMF(2+); DMF = N,N dimethylformamide) in a tetrahydrofuran (THF) solution containing excess triethylamine (NEt(3)). The mechanism of the Ru-H(+) formation was investigated in detail. A photochemical ligand substitution reaction of Ru-DMF(2+) in THF proceeded to give [Ru(tpy)(bpy)(THF)](2+) (Ru-THF(2+)) with a quantum yield of (7.6 +/- 0.7) x 10(-2). In the presence of NEt(3), a similar photochemical ligand substitution reaction also proceeded quickly, but the products were an equilibrium mixture of Ru-THF(2+) and [Ru(tpy)(bpy)(NEt(3))](2+) (Ru-NEt(3)(2+)) with a considerable amount of Ru-H(+) even in the first stage of the photochemical reaction. The equilibrium constant between Ru-THF(2+) and Ru NEt(3)(2+) was determined as 6.9 +/- 2.1. Irradiation to Ru-NEt(3)(2+) gave Ru H(+) with a quantum yield of (9.1 +/- 0.5) x 10(-3). An important intermediate, Ru-NEt(3)(2+), was isolated, and its properties were investigated in detail. PMID- 19810722 TI - Phase space prediction of product branching ratios: canonical competitive nonstatistical model. AB - We present a new model for predicting branching ratios of chemical reactions when a branching of the reaction path occurs after the dynamical bottleneck, including the case where it occurs after an intermediate. The model is based on combining nonstatistical phase space theory for the direct component of a reaction with variational transition-state theory for an indirect component of reaction. The competition between direct and indirect processes is treated by an extension of the unified statistical model. This new method provides a way to understand the factors that control this kind of chemical reaction and to perform calculations using high-level electronic structure methods for complex systems. The model is based on quantized energy levels of transition states and products, and it involves the same information as required for calculating transition-state rate constants and equilibrium constants plus a phenomenological relaxation time, which was taken from previous work. For the textbook reaction of the hydroboration of propene by BH(3) it has recently been inferred that the selectivity can only be understood by consideration of dynamical trajectories. However, the calculated branching fraction of this prototype reaction increases from 2%-3% when calculated under the inappropriate assumption of complete equilibration of the intermediate to from 8%-9% when calculated with the new theory, which requires only limited information about the system and does not involve running trajectories. The calculated result is in reasonable agreement with experiment (approximately 10%). PMID- 19810723 TI - Wrinkled, dual-scale structures of diamond-like carbon (DLC) for superhydrophobicity. AB - We present a simple two-step method to fabricate dual-scale superhydrophobic surfaces by using replica molding of poly(dimethylsiloxane) (PDMS) micropillars, followed by deposition of a thin, hard coating layer of a SiO(x)-incorporated diamond-like carbon (DLC). The resulting surface consists of microscale PDMS pillars covered by nanoscale wrinkles that are induced by residual compressive stress of the DLC coating and a difference in elastic moduli between DLC and PDMS without any external stretching or thermal contraction on the PDMS substrate. We show that the surface exhibits superhydrophobic properties with a static contact angle over 160 degrees for micropillar spacing ratios (interpillar gap divided by diameter) less than 4. A transition of the wetting angle to approximately 130 degrees occurs for larger spacing ratios, changing the wetting from a Cassie Cassie state (C(m)-C(n)) to a Wenzel-Cassie state (W(m)-C(n)), where m and n denote micro- and nanoscale roughness, respectively. The robust superhydrophobicity of the Cassie-Cassie state is attributed to stability of the Cassie state on the nanoscale wrinkle structures of the hydrophobic DLC coating, which is further explained by a simple mathematical theory on wetting states with decoupling of nano- and microscale roughness in dual scale structures. PMID- 19810724 TI - STM investigation of temperature-dependent two-dimensional supramolecular architectures of C60 and amino-tetraphenylporphyrin on Ag(110). AB - Multicomponent supramolecular self-assemblies of exceptional long-range order and low defectivity are obtained if C(60) and 5-(4-aminophenyl)-10,15,20 triphenylporphyrin (TPP-NH2) are assembled on Ag(110) by sequential evaporation in the submonolayer range of TPP-NH2 and fullerene on the substrate surface and subsequent annealing. A (+/-2 -3, 6 +/- 3) array consisting of supramolecular stripes of a 1:1 C(60)/TPP-NH2 2D adduct develops at 410 K (the low temperature, LT, phase). If the LT phase is annealed at 470 K, then a 3:1 fullerene/TPP-NH2 (+/-3 -5, 5 +/- 5) nanoporous array (the HT phase) forms, with each pore containing a single porphyrin molecule. Phase separation occurs by annealing the HT phase at 520 K. Structural models are proposed and discussed on the basis of the experimental scanning tunneling microscopy results. PMID- 19810725 TI - Surface modification of upconverting NaYF4 nanoparticles with PEG-phosphate ligands for NIR (800 nm) biolabeling within the biological window. AB - We present a technique for the replacement of oleate with a PEG-phosphate ligand [PEG = poly(ethylene glycol)] as an efficient method for the generation of water dispersible NaYF(4) nanoparticles (NPs). The PEG-phosphate ligands are shown to exchange with the original oleate ligands on the surface of the NPs, resulting in water-dispersible NPs. The upconversion intensity of the NPs in aqueous environments was found to be severely quenched when compared to the original NPs in organic solvents. This is attributed to an increase in the multiphonon relaxations of the lanthanide excited state in aqueous environments due to high energy vibrational modes of water molecules. This problem could be overcome partially by the synthesis of core/shell NPs which demonstrated improved photophysical properties in water over the original core NPs. The PEG-phosphate coated upconverting NPs were then used to image a line of ovarian cancer cells (CaOV3) to demonstrate their promise in biological application. PMID- 19810727 TI - Exciton-dissociation and charge-recombination processes in pentacene/C60 solar cells: theoretical insight into the impact of interface geometry. AB - The exciton-dissociation and charge-recombination processes in organic solar cells based on pentacene/C(60) heterojunctions are investigated by means of quantum-mechanical calculations. The electronic couplings and the rates of exciton dissociation and charge recombination have been evaluated for several geometrical configurations of the pentacene/C(60) complex, which are relevant to bilayer and bulk heterojunctions. The results suggest that, irrespective of the actual pentacene-fullerene orientation, both pentacene-based and C(60)-based excitons are able to dissociate efficiently. Also, in the case of parallel configurations of the molecules at the pentacene/C(60) interface, the decay of the lowest charge-transfer state to the ground state is calculated to be very fast; as a result, it can compete with the dissociation process into mobile charge carriers. Since parallel configurations are expected to be found more frequently in bulk heterojunctions than in bilayer heterojunctions, the performance of pentacene/C(60) bulk-heterojunction solar cells is likely to be more affected by charge recombination than that of bilayer devices. PMID- 19810726 TI - Structural and functional studies of Aspergillus oryzae cutinase: enhanced thermostability and hydrolytic activity of synthetic ester and polyester degradation. AB - Cutinases are responsible for hydrolysis of the protective cutin lipid polyester matrix in plants and thus have been exploited for hydrolysis of small molecule esters and polyesters. Here we explore the reactivity, stability, and structure of Aspergillus oryzae cutinase and compare it to the well-studied enzyme from Fusarium solani. Two critical differences are highlighted in the crystallographic analysis of the A. oryzae structure: (i) an additional disulfide bond and (ii) a topologically favored catalytic triad with a continuous and deep groove. These structural features of A. oryzae cutinase are proposed to result in an improved hydrolytic activity and altered substrate specificity profile, enhanced thermostability, and remarkable reactivity toward the degradation of the synthetic polyester polycaprolactone. The results presented here provide insight into engineering new cutinase-inspired biocatalysts with tailor-made properties. PMID- 19810728 TI - Incomplete exciton harvesting from fullerenes in bulk heterojunction solar cells. AB - We investigate the internal quantum efficiencies (IQEs) of high efficiency poly-3 hexylthiophene:[6,6]-phenyl-C(61)-butyric acid methyl ester (P3HT:PCBM) solar cells and find them to be lower at wavelengths where the PCBM absorbs. Because the exciton diffusion length in PCBM is too small, excitons generated in PCBM decay before reaching the donor-acceptor interface. This result has implications for most state of the art organic solar cells, since all of the most efficient devices use fullerenes as electron acceptors. PMID- 19810729 TI - Electrogenerated IrO(x) nanoparticles as dissolved redox catalysts for water oxidation. AB - We describe the first example of redox catalysis using a dissolved electroactive nanoparticle, based on the oxidation of water by electrogenerated IrO(x) nanoparticles containing Ir(VI) states, in pH 13 solutions of 1.6 +/- 0.6 nm (dia.) Ir(IV)O(x) nanoparticles capped solely by hydroxide. At potentials (ca. +0.45 V) higher than the mass transport-controlled plateau of the nanoparticle Ir(V/IV) wave, rising large redox catalytic currents reflect electrochemical generation of Ir(VI) states, which by +0.55 V and onward to +1.0 V are shown by rotated ring disk electrode experiments to lead with 100% current efficiency to the oxidation of water to O(2). O(2) production at +0.55 V corresponds to an overpotential eta of only 0.29 V, relative to thermodynamic expectations of the four electron H(2)O-->O(2) reaction. The Ir site turnover frequency (TO, mol O(2)/Ir sites/s) is 8-11 s(-1). Controlled potential coulometry shows that all Ir sites in these nanoparticles (average 66 Ir each) are electroactive, meaning that the nanoparticles are small enough to allow the required electron and proton transport throughout. Both the overpotential and TO values are nearly the same as those observed previously for films electroflocculated from similar IrO(x) nanoparticles, providing the first comparison of electrocatalysis by nanoparticle films with redox catalysis by dissolved, diffusing nanoparticles. PMID- 19810730 TI - Virtual high throughput screening confirmed experimentally: porous coordination polymer hydration. AB - Hydrothermal stability is a pertinent issue to address for many industrial applications where percent levels of water can be present at temperatures ranging from subambient to several hundred degrees. Our objective is to understand relative stabilities of MOF materials through experimental testing combined with molecular modeling. This will enable the ultimate design of materials with improved hydrothermal stability, while maintaining the properties of interest. The tools that we have employed for these studies include quantum mechanical calculations based upon cluster models and combinatorial steaming methods whereby a steam stability map was formulated according to the relative stability of different materials. The experimental steaming method allows for high throughput screening of materials stability over a broad range of steam levels as well as in depth investigation of structural transformations under more highly resolved conditions, while the cluster model presented here yields the correct trends in hydrothermal stability. Good agreement was observed between predicted relative stabilities of materials by molecular modeling and experimental results. Fundamental information from these studies has provided insight into how metal composition and coordination, chemical functionality of organic linker, framework dimensionality, and interpenetration affect the relative stabilities of PCP materials. This work suggests that the strength of the bond between the metal oxide cluster and the bridging linker is important in determining the hydrothermal stability of the PCP. Although the flexibility of the framework plays a role, it is not as important as the metal-linker bond strength. This demonstration of alignment between experimental and calculated observations has proven the validity of the method, and the insight derived herein insight facilitates direction in designing ideal MOF materials with improved hydrothermal stability for desired applications. PMID- 19810732 TI - Hydrogen storage properties of nanosized MgH2-0.1TiH2 prepared by ultrahigh energy-high-pressure milling. AB - Magnesium hydride (MgH(2)) is an attractive candidate for solid-state hydrogen storage applications. To improve the kinetics and thermodynamic properties of MgH(2) during dehydrogenation-rehydrogenation cycles, a nanostructured MgH(2) 0.1TiH(2) material system prepared by ultrahigh-energy-high-pressure mechanical milling was investigated. High-resolution transmission electron microscope (TEM) and scanning TEM analysis showed that the grain size of the milled MgH(2) 0.1TiH(2) powder is approximately 5-10 nm with uniform distributions of TiH(2) among MgH(2) particles. Pressure-composition-temperature (PCT) analysis demonstrated that both the nanosize and the addition of TiH(2) contributed to the significant improvement of the kinetics of dehydrogenation and hydrogenation compared to commercial MgH(2). More importantly, PCT cycle analysis demonstrated that the MgH(2)-0.1TiH(2) material system showed excellent cycle stability. The results also showed that the DeltaH value for the dehydrogenation of nanostructured MgH(2)-0.1TiH(2) is significantly lower than that of commercial MgH(2). However, the DeltaS value of the reaction was also lower, which results in minimum net effects of the nanosize and the addition of TiH(2) on the equilibrium pressure of dehydrogenation reaction of MgH(2). PMID- 19810731 TI - Synthesis and biochemical analysis of complex chain-elongation intermediates for interrogation of molecular specificity in the erythromycin and pikromycin polyketide synthases. AB - The 6-deoxyerythronolide B synthase (DEBS) and pikromycin (Pik) polyketide synthase (PKS) are unique multifunctional enzyme systems that are responsible for the biosynthesis of the erythromycin and pikromycin 14-membered ring aglycones, respectively. Together, these natural product biosynthetic systems provide excellent platforms to examine the fundamental structural and catalytic elements that govern polyketide assembly, processing, and macrocyclization. In these studies, the native pentaketide intermediate for DEBS was synthesized and employed for in vitro chemoenzymatic synthesis of macrolactone products in engineered monomodules Ery5, Ery5-TE, and Ery6. A comparative analysis was performed with the corresponding Pik module 5 (PikAIII) and module 6 (PikAIV), dissecting key similarities and differences between these highly related PKSs. The data revealed that individual modules in the DEBS and Pik PKSs possess distinctive molecular selectivity profiles and suggest that substrate recognition has evolved unique characteristics in each system. PMID- 19810733 TI - Chemical modification of the insecticidal briantheins X and Y. AB - Manipulation of the allylic chloride functionality in briantheins X and Y provided 10 new analogues of these gorgonian diterpenes as part of a continuing study of structure-activity relationships in this family of insecticidal compounds. Modified Finkelstein reaction conditions led not only to halogen substitution products but also to rearrangement, dehydrohalogenation, and dehydration products. None of the new compounds showed superior insecticidal activity to brianthein X or Y, although most did result in lower weight gains versus controls. PMID- 19810735 TI - Lanthanide germanate cluster organic frameworks constructed from {Ln(8)Ge(12)} or {Ln(11)Ge(12)} cage cluster building blocks. AB - Through the use of bis(carboxyethylgermanium) sesquioxide as the germanium source, lanthanide has been successfully introduced into germanate systems under hydrothermal conditions, resulting in a novel lanthanide germanate cluster organic framework with twofold-interpenetrating nets built from Nd(8)Ge(12) cluster units. To avoid the interpenetration, a second ligand, 2-picolinic acid, was introduced into the above system, resulting in the formation of a series of noninterpenetrating networks constructed from Ln(11)Ge(12) cluster units, as expected. PMID- 19810734 TI - Nanoassembled capsules as delivery vehicles for large payloads of high relaxivity Gd3+ agents. AB - Nanoassembled capsules (NACs) that incorporate a polymer aggregate inside a silica shell may be loaded with agents that are of particular interest for therapeutic or diagnostic applications. NACs formed using the MRI contrast agent GdDOTP(5-) in the internal polymer aggregate are reported herein, the smaller of which show promise as potential MRI contrast agents. Unlike many other nanoencapsulated systems, water access to the inner core of these NACs does not appear to be limited and consequently the water relaxivity per Gd(3+) agent can reach as high as 24 mM(-1) s(-1). Robust, spherical capsules were formed using polyallylamine or poly-L-lysine ranging from 0.2 to 5 microm in diameter. The greatest gains in relaxivity were observed for smaller NACs, for which water accessibility remained high but molecular rotation of the Gd(3+) chelate was effectively restricted. Larger NACs did not afford such large gains in relaxivity, the result of poorer water accessibility combined with less-effective rotational restriction. PMID- 19810736 TI - Synthesis and characterization of biodegradable poly(ester amide)s with pendant amine functional groups and in vitro cellular response. AB - The purpose of this study was to use a convenient synthetic strategy to prepare a new family of biodegradable amino acid-based poly(ester amide)s (PEAs) with pendant amine groups along the polymer backbone, and investigate the applications of the new polymers in the biomedical area. Two amino acids, L-phenylalanine (Phe) and L-lysine (Lys), were used as the model amino acid compounds to illustrate the synthesis, characterization, and biological property of this new family of functional PEAs. These new PEAs were obtained by two-step reactions, the ring-opening reaction of epsilon-(benzyloxycarbonyl)-L-lysine N carboxyanhydride (Z-LysNCA) with L-phenylalanine hexane-1,6-diol diester p toluenesulfonate (Phe-6), and subsequently solution polycondensation with di-p nitrophenyl sebacoyl (NS). The benzyloxycarbonyl (Z) protective groups of the resulting polymer (PEA-Z-Lys) were completely removed to produce the new functional PEAs having free pendant amine groups (PEA-Lys-NH(2)). The level of the pendant amine groups on the PEA-Lys-NH(2) could be tailored by adjusting the Phe-6 to Z-LysNCA feed ratio. Analyses of FTIR, (1)H NMR, (13)C NMR spectra, and DSC revealed the desired chemical structures and thermal property of PEA-Z-Lys as well as the final functional PEA-Lys-NH(2). The free pendant amine groups were used to chemically conjugate a fluorescent dye to demonstrate the utility of this new family of functional PEA. An in vitro cell culture study of these functional PEAs showed that they supported the proliferation of bovine aortic endothelial cell slightly better than gelatin-coated glass coverslips. This new family of biodegradable functional PEA with free amine groups may have great potential applications for biomedical and pharmacological fields. PMID- 19810737 TI - How the substituent at O-3 of N-acetylglucosamine impacts glycosylation at O-4: a comparative study. AB - An assessment of the relative reactivities of the 4-OH of N-acetylglucosamine acceptors bearing simple protecting groups, beta-linked or alpha-linked D or L sugars at O-3 is presented, using a per-O-acetylated alpha-D-glucosyl trichloroacetimidate donor under activation by BF(3) x OEt(2). The presence of either an acyl or carbonate protecting group at O-3 did not impact the reactivity at O-4 with all glycosylations proceeding successfully. On the other hand, the presence of peracetylated sugars at O-3 of N-acetylglucosamine acceptors did impact the reactivity of the 4-OH. The acceptors with an alpha-D-Man, beta-D-Gal, or beta-D-Glc at O-3 reacted promptly. In comparison, the acceptors bearing a beta-L-Fuc, alpha-L-Fuc, or alpha-L-Rha underwent glucosylation slowly, and unreacted acceptor was recovered from the reaction mixtures. Systematic searches carried out on the disaccharide acceptors and trisaccharide products carrying either a peracetylated beta-D-Gal or beta-L-Fuc at O-3 of the glucosamine residue suggest that, for these two acceptors, a conformational reorientation necessary around the fucosidic linkage contributes to the lower reactivity of the beta fucosylated acceptor. The acceptors bearing a beta-linked D-Gal, D-Glc, or L-Fuc residue at O-3 each gave trisaccharide products that were mostly stable in the reaction conditions. In contrast, the alpha-linked residues at O-3 were rather unstable in these reaction conditions and the degradation of either the acceptors or trisaccharide products led to low glycosylation yields. In these later reactions, it was impossible to clearly assess which of the acceptor or product underwent degradation as comigration and detection issues prevented us from following these glycosylations by TLC or RP-HPLC. In contrast, the glycosylation of an acceptor carrying an alpha-linked perbenzylated L-Fuc residue at O-3 could be easily monitored by RP-HPLC. The data obtained when monitoring this glycosylation showed that the acceptor underwent prompt glycosylation but a decrease in the absorbance peak corresponding to the trisaccharide along with the appearance of a peak corresponding to a perbenzylated fucose hemiacetal indicated that the trisaccharide product was unstable in the reaction conditions. PMID- 19810738 TI - Kinetics of oxygen adsorption and initial oxidation on Cu(110) by hyperthermal oxygen molecular beams. AB - Oxygen adsorption and subsequent oxide formation on Cu(110) using a hyperthermal oxygen molecular beam (HOMB) were investigated using X-ray photoelectron spectrometry. The O-uptake curves, which were determined from the evolution of the O-1s peaks, indicate that simple Langmuir type kinetics can describe dissociative adsorption of O(2) with an incident energy (E(i)) below 0.5 eV under Theta < or = 0.5 ML. The reaction order dependence on E(i) implies two competing dissociation mechanisms, trapping-mediated and directly activated adsorption. Oxidation at Theta > or = 0.5 ML proceeds rather effectively using highly energetic HOMB at E(i) > or = 1.0 eV. The azimuthal dependence of the sticking probability during the effective oxidation using HOMB incidence suggests that the added rows, which consist of the Cu-O structure, shade the reactive hollow sites in the trough where oxygen penetrates into the subsurface. The surface Cu(2)O formed with highly energetic HOMB incidence decomposes with desorbing subsurface oxygen even at room temperature, demonstrating that HOMB can induce a metastable surface structure that cannot be produced in the thermal equilibrium process. PMID- 19810739 TI - Low density lipoprotein detection based on antibody immobilized self-assembled monolayer: investigations of kinetic and thermodynamic properties. AB - Human plasma low density lipoprotein (LDL) immunosensor based on surface plasmon resonance (SPR) and quartz crystal microbalance (QCM) was fabricated by immobilizing antiapolipoprotein B (AAB) onto self-assembled monolayer (SAM) of 4 aminothiophenol (ATP). The AAB/ATP/Au immunosensor can detect LDL up to 0.252 microM (84 mg/dL) and 0.360 microM (120 mg/dL) with QCM and SPR, respectively. The SPR and QCM measurements were further utilized to study the reaction kinetics of the AAB-LDL interaction. The adsorption process involved was explored using Langmuir adsorption isotherm and Freundlich adsorption models. The thermodynamic parameters such as change in Gibb's free energy (DeltaG(ads)), change in enthalpy (DeltaH(ads)), and change in entropy (DeltaS(ads)) determined at 283, 298, and 308 K revealed that the AAB-LDL interaction is endothermic in nature and is governed by entropy. Kinetic, thermodynamic, and sticking probability studies disclosed that desorption of the water molecules from the active sites of AAB and LDL plays a key role in the interaction process and increase in temperature favors binding of LDL with the AAB/ATP/Au immunosensor. Thus, the studies were utilized to unravel the most important subprocess involved in the adsorption of LDL onto AAB-modified ATP/Au surface that may help in the fabrication of LDL immunosensors with better efficiency. PMID- 19810740 TI - Quantum chemical calculation of type-1 cu reduction potential: ligand interaction and solvation effect. AB - Using active site model molecules consisting of approximately 100 atoms, the reduction potentials of five type-1 Cu centers in cucumber stellacyanin, fern dryopteris crassirhizoma plastocyanin, Met148Gln rusticyanin, wild type rusticyanin, and Met148Leu rusticyanin were calculated with a heterogeneous conductor-like polarizable continuum model and the B3LYP/6-311++G(2df,p) method. The results are 242, 366, 522, 667, and 825 mV, respectively, in good agreement with experimental values 260, 376, 563, 667, and 798 mV. Ligand interaction ( approximately 250 mV) and solvation effect ( approximately 250 mV) are found to be the main determinants of the relative E(0) of these five type-1 Cu centers. PMID- 19810741 TI - Modification of coal as a fuel for the direct carbon fuel cell. AB - As a promising high-temperature fuel cell, the direct carbon fuel cell (DCFC) has a much higher efficiency and a lower emission as compared with conventional coal fired power plants. To develop an increased understanding of the relationship between the microstructure, surface chemistry, and electrochemical performance of coal as a fuel for the DCFC, a coal sample from Central Queensland has been subjected to various pretreatments, including acid washing, air oxidation, and pyrolysis. It has been found that an acid treatment of the coal enhanced its electrochemical reactivity due to an increase in oxygen-containing surface functional groups. By contrast, heat treatment of the coal results in a sharp decrease in the electrochemical reactivity in the DCFC due to a decrease in the oxygen-containing surface functional groups, particularly CO(2)-yielding surface groups. A higher surface area of coal may also be helpful, but much less important than surface chemistry. PMID- 19810742 TI - Generalized indirect covariance NMR formalism for establishment of multidimensional spin correlations. AB - Multidimensional nuclear magnetic resonance (NMR) experiments measure spin-spin correlations, which provide important information about bond connectivities and molecular structure. However, direct observation of certain kinds of correlations can be very time-consuming due to limitations in sensitivity and resolution. Covariance NMR derives correlations between spins via the calculation of a (symmetric) covariance matrix, from which a matrix-square root produces a spectrum with enhanced resolution. Recently, the covariance concept has been adopted to the reconstruction of nonsymmetric spectra from pairs of 2D spectra that have a frequency dimension in common. Since the unsymmetric covariance NMR procedure lacks the matrix-square root step, it does not suppress relay effects and thereby may generate false positive signals due to chemical shift degeneracy. A generalized covariance formalism is presented here that embeds unsymmetric covariance processing within the context of the regular covariance transform. It permits the construction of unsymmetric covariance NMR spectra subjected to arbitrary matrix functions, such as the square root, with improved spectral properties. This formalism extends the domain of covariance NMR to include the reconstruction of nonsymmetric NMR spectra at resolutions or sensitivities that are superior to the ones achievable by direct measurements. PMID- 19810743 TI - Denaturation resistance of beta-lactoglobulin in monomolecular films at the air water interface. AB - Using X-ray reflectometry we report strong differences in the denaturation response of beta-lactoglobulin adsorbed as a monomolecular film at the air-water interface from that observed in mixed denaturant/beta-lactoglobulin bulk solutions. Using the "flow trough" technique an isolated monomolecular film of the protein showed little change in structure when subjected to a 4.0 M guanidinium hydrochloride substrate. Unlike the bulk solution where a new protein layer structure appears, small changes in the protein packing and the roughness of the film are the only evidence of change. These parameters have been studied as a function of denaturant concentration and film quality. The strength of the response depends on the degree of perfection of the originally formed film; quickly formed films are more easily denatured. As the response is so subtle, possible interfering effects such as denaturant release of protein adsorbed on the trough have been quantified. PMID- 19810744 TI - Stepwise hydration of protonated carbonic acid: a theoretical study. AB - The gas-phase geometries, binding energies (BEs), and sequential binding energies (SBEs) of protonated carbonic acid (PCA)-water (W) clusters (PCAW(n), where n = 1 6) have been calculated using density functional theory (DFT) with Becke's three parameter hybrid exchange functional and the Lee-Yang-Parr correlation functional (B3LYP) and M05-2X methods. The presence of wirelike structures of protonated water in PCAW(n)(x) clusters is evident from the results. The results indicate that a proton is transferred from PCA to its immediate water molecule in the linear and monohydroxy clusters of PCA. The involvement of the Eigen cation and Grotthuss type of mechanism in the proton transport is observed from the sequential hydration energies and from the calculated vibrational spectra. Although geometrical parameters clearly reveal the presence of the Eigen core, calculated lower-energy vibrational modes provide clues about the involvement of the sequence Eigen --> Zundel --> Eigen in the proton transfer. PMID- 19810745 TI - Enantioselective HF loss promoted by resonant two-photon ionization of supersonically expanded (R)-1-phenyl-2,2,2-trifluoroethanol clusters. AB - (R)-1-phenyl-2,2,2-trifluoroethanol and its hydrogen bonded adducts with achiral (water, tetrahydrofuran) and chiral solvent molecules ((R)- and (S)-butan-2-ol, (R)- and (S)-3-hydroxy-tetrahydrofuran) have been ionized by resonant two-photon absorption. The presence of photofragments, attributable to the occurrence of a hydrogen fluoride loss reaction, has been interpreted with the aid of theoretical predictions at the DFT level of theory with the inclusion of dispersive terms. The HF elimination process takes place by a mechanism involving the preliminary C(alpha)-H hydrogen transfer to an hydroxyl oxygen assisted by the solvent molecule which is followed by extrusion of the HF molecule. The calculated energy barriers depend on the type of solvent as well as on its configuration and are consistent with the observed fragmentation ratios. PMID- 19810746 TI - Erythrocyte-endothelium adhesion can be induced by dextran. AB - Plasma proteins have been identified to play a key role in the increased adhesiveness of red blood cells (RBC) to endothelial cells (EC) in various diseases associated with vascular complications. However, the underlying mechanisms on how plasma proteins facilitate adhesion remain unclear. In this study, we investigated if macromolecular depletion is able to induce adhesion of RBC to EC. RBC were suspended in solutions containing the neutral polyglucose dextran to mimic the effects of nonadsorbing macromolecules and the dynamics of RBC-EC adhesion were recorded using a parallel plate flow chamber system. Cell adhesion was markedly increased in the presence of dextran with a molecular mass larger than 70 kDa, with this increase reflected by both the number of cells adhering and the strength of the adhesion. This increased adhesiveness is attributed to reduced surface concentrations of the large polymers and hence attractive forces due to depletion interaction. Our results thus provide a rational explanation on how nonadsorbing plasma proteins could play a significant role in abnormal RBC-EC adhesion in vivo. PMID- 19810747 TI - Hydrogenolysis of unstrained carbon-carbon sigma bonds: stereoselective entry into benzylic tertiary centers. AB - The modification of sp(3)-hybridized carbon centers through Pd-catalyzed reductive cleavage of unstrained carbon-carbon sigma bonds is described. From the hydrogenolysis of benzyl Meldrum's acids bearing an all-carbon benzylic quaternary center, Meldrum's acid and aromatics substituted with a tertiary benzylic stereocenter were obtained in good to excellent yields. Mechanistic studies showed that the reductive cleavage of enantioenriched benzylic quaternary centers proceeded with inversion of configuration, supporting a "loose" S(N)2 pathway. PMID- 19810748 TI - Thermoelectric effect across the metal-insulator domain walls in VO2 microbeams. AB - We report on measurements of Seebeck effect in single-crystal VO(2) microbeams across their metal-insulator phase transition. One-dimensionally aligned metal insulator domain walls were reversibly created and eliminated along single VO(2) beams by varying temperature, which allows for accurate extraction of the net contribution to the Seebeck effect from these domain walls. We observed significantly lower Seebeck coefficient in the metal-insulator coexisting regime than predicted by a linear combination of contributions from the insulator and metal domains. This indicates that the net contribution of the domain walls has an opposite sign from that of the insulator and metal phases separately. Possible origins that may be responsible for this unexpected effect were discussed in the context of complications in this correlated electron material. PMID- 19810749 TI - Metal speciation dynamics in dispersions of soft colloidal ligand particles under steady-state laminar flow condition. AB - A theory is presented for metal speciation dynamics in a swarm of soft, spherical core-hell colloidal ligand particles under steady-state laminar flow condition. Mass transfer and subsequent complexation of metal species within the reactive, permeable particle shell are governed by the interplay between (i) convective diffusion of free metal ions M within and around the shell where ligands L are distributed, and (ii) kinetics of ML complex formation/dissociation in the shell. The local concentrations of metal M and complex ML are determined by the convective-diffusion equations with appropriate chemical source term and full account of radial and angular concentration polarization contributions. The steady-state flow field is determined from the solution of Navier-tokes equation including convective acceleration term for the fluid external to the particle, and from Brinkman equation for the internal fluid flow. The confined location of ligands within the particle shell leads to ML formation/dissociation rate constants (denoted as ka* and kd*, respectively) that differ significantly from their counterparts (ka and kd) defined for homogeneous ligand distribution throughout the solution. The relationship between ka,d* and ka,d is derived from the numerical evaluation of the spatial, time-dependent distributions of free and bound metal within and/or outside the particle. The dramatic dependence of ka,d* on hydrodynamic particle softness, Peclet number, soft surface layer thickness, and particle radius are analyzed in the steady-state nonequilibrium chemical regime within the context of dynamic features for colloidal complexes. The analysis covers the limiting cases of hydrodynamically impermeable, hard particles where binding sites are located at the very surface of the particle core (e.g., functionalized latex colloids) and free draining, polymeric ligand particles devoid of a hard core (e.g., porous gel particles). The formalism further applies to any values of the Peclet number, that is, for speciation dynamics determined by kinetic processes coupled to diffusion and/or convection metal mass transfer(s). A discussion is provided for the comparison between the exact numerical results and the analytical formulation based on the approximate Levich expression for convective-diffusion metal flux at the surface of hard ligand particles. PMID- 19810750 TI - Sortase activity is controlled by a flexible lid in the pilus biogenesis mechanism of gram-positive pathogens. AB - Pili are surface-linked virulence factors that play key roles in infection establishment in a variety of pathogenic species. In Gram-positive pathogens, pilus formation requires the action of sortases, dedicated transpeptidases that covalently associate pilus building blocks. In Streptococcus pneumoniae, a major human pathogen, all genes required for pilus formation are harbored in a single pathogenicity islet which encodes three structural proteins (RrgA, RrgB, RrgC) and three sortases (SrtC-1, SrtC-2, SrtC-3). RrgB forms the backbone of the streptococcal pilus, to which minor pilins RrgA and RrgC are covalently associated. SrtC-1 is the main sortase involved in polymerization of the RrgB fiber and displays a lid which encapsulates the active site, a feature present in all pilus-related sortases. In this work, we show that catalysis by SrtC-1 proceeds through a catalytic triad constituted of His, Arg, and Cys and that lid instability affects protein fold and catalysis. In addition, we show by thermal shift analysis that lid flexibility can be stabilized by the addition of substrate-like peptides, a feature shared by other periplasmic transpeptidases. We also report the characterization of a trapped acyl-enzyme intermediate formed between SrtC-1 and RrgB. The presence of lid-encapsulated sortases in the pilus biogenesis systems in many Gram-positive pathogens points to a common mechanism of substrate recognition and catalysis that should be taken into consideration in the development of sortase inhibitors. PMID- 19810751 TI - Picosecond fluorescence relaxation spectroscopy of the calcium-discharged photoproteins aequorin and obelin. AB - Addition of calcium ions to the Ca(2+)-regulated photoproteins, such as aequorin and obelin, produces a blue bioluminescence originating from a fluorescence transition of the protein-bound product, coelenteramide. The kinetics of several transient fluorescent species of the bound coelenteramide is resolved after picosecond-laser excitation and streak camera detection. The initially formed spectral distributions at picosecond-times are broad, evidently comprised of two contributions, one at higher energy (approximately 25,000 cm(-1)) assigned as from the Ca(2+)-discharged photoprotein-bound coelenteramide in its neutral state. This component decays much more rapidly (t(1/2) approximately 2 ps) in the case of the Ca(2+)-discharged obelin than aequorin (t(1/2) approximately 30 ps). The second component at lower energy shows several intermediates in the 150-500 ps times, with a final species having spectral maxima 19 400 cm(-1), bound to Ca(2+)-discharged obelin, and 21 300 cm(-1), bound to Ca(2+)-discharged aequorin, and both have a fluorescence decay lifetime of 4 ns. It is proposed that the rapid kinetics of these fluorescence transients on the picosecond time scale, correspond to times for relaxation of the protein structural environment of the binding cavity. PMID- 19810752 TI - Identification and characterization of binding sites on S100A7, a participant in cancer and inflammation pathways. AB - S100A7 (psoriasin) is a member of the S100 family of signaling proteins. It is implicated in and considered a therapeutic target for inflammation and cancer, yet no small molecule ligands for S100A7 have been identified. To begin the development of specific small molecule inhibitors of S100A7 function, we have used a series of surface binding fluorescent dyes to probe the surface hydrophobic sites. Two naphthalene-based dyes (2,6-ANS and 1,8-ANS) were found to bind S100A7 in a distinct cleft. We characterized the binding interaction by determining both the structure of S100A7 bound to 2,6-ANS and the structure of S100A7 bound to 1,8-ANS to 1.6 A. In both cases, two molecules of dye were docked such that the naphthalene groups were positioned in two symmetry-related grooves that are formed by the N-terminal helices of each monomer. We observed that Met12 acts as a gatekeeper to the binding cleft, adopting an "open" conformation for the more elongated 2,6-ANS while remaining in a "closed" conformation for the more compact 1,8-ANS. Steady-state fluorescence experiments revealed that S100A7 binds two copies of 2,6-ANS, each with a K(d) of 125 muM. Time-resolved fluorescence lifetime measurements indicated that the two molecules of 2,6-ANS bind in two independent binding sites with different fluorescence lifetimes, suggesting that the S100A7 homodimer is not perfectly symmetric in solution. Isothermal titration calorimetry studies demonstrate that S100A7 has a higher affinity for 2,6-ANS than 1,8-ANS. Yeast two-hybrid studies were also used to probe contributions of individual residues of an S100A7 triple mutant with respect to Jab1 binding. Mutation of Leu78, which forms part of the Met12 cleft occupied by 2,6-ANS, reduced the level of Jab1 binding, suggesting a potentially important role for the Met12 hydrophobic pocket in defining a Jab1 interface. Additional Y2H studies also delineate contributions of Gln88 and in particular Asp56 that shows the most significant abrogated binding to Jab1. Collectively, these data suggest a complex interaction between S100A7 and the much larger Jab1. These studies form the basis for the development of small molecule reporters and modifiers of S100A7 form and function. PMID- 19810753 TI - Multipotent adult germline stem cells and embryonic stem cells: comparative proteomic approach. AB - Spermatogonial stem cells isolated from the adult mouse testis acquire under certain culture conditions pluripotency and become so-called multipotent adult germline stem cells (maGSCs). They can be differentiated into somatic cells of the three germ layers. We investigated a subset of the maGSCs and ESCs proteomes using cell lines derived from two different mouse strains, narrow range immobilized pH gradients to favor the detection of less abundant proteins, and DIGE to ensure confident comparison between the two cell types. 2-D reference maps of maGSCs and ESCs in the pI ranges 3-6 and 5-8 were created, and protein entities were further processed for protein identification. By peptide mass fingerprinting and tandem mass spectrometry combined with searches of protein sequence databases, a set of 409 proteins was identified, corresponding to a library of 166 nonredundant stem cell-associated proteins. The identified proteins were classified according to their main known/postulated functions using bioinformatics. Furthermore, we used DIGE to highlight the ESC-like nature of maGSCs on the proteome scale. We concluded that the proteome of maGSCs is highly similar to that of ESCs as we could identify only a small subset of 18 proteins to be differentially expressed between the two cell types. Moreover, comparative analysis of the cell line proteomes from two different mouse strains showed that the interindividual differences in maGSCs proteomes are minimal. With our study, we created for the first time a proteomic map for maGSCs and compared it to the ESCs proteome from the same mouse. We confirmed on the proteome level the ESC like nature of maGSCs. PMID- 19810755 TI - Thermoreversible gelation of poly(vinylidene fluoride-co-hexafluoro propylene) in phthalates. AB - The thermoreversible gelation of poly(vinylidene fluoride-co-hexafluoro propylene) copolymer have been studied in a series of phthalates, Ph-(COO C(n)H(2n+1))(2) with n = 1-8. The gelation rate increases with increasing aliphatic chain length up to n = 6, and the gelation phenomena does not occur for higher n > 6. The fibrillar morphology is evident for dried gels whose dimension (both lateral and thickness) becomes shorter and thinner with increasing n. The structures of the gels formed in various phthalates have been investigated by small-angle neutron scattering and small-angle X-ray scattering techniques, suggesting sheet-like structure, where the interplanner distance increases with increasing aliphatic chain length. The scattering intensity I(q) decreases with q according to the Ornstein-Zernike model, where q = (4pi/lambda) sin theta (2theta and lambda are scattering angle and wavelength of neutron) and the correlation length, xi, assigned to the average distance between the neighboring crystallites, also increases with increasing aliphatic chain length of diesters. The detailed thermal analyses and phase diagrams of the copolymer gels have been studied in a wide range of phthalates. Further, polymer-solvent complexes leading to the formation of two distinct compounds have been reported. A systematic change of compound composition has also been observed in the whole range of phthalates studied here. On the basis of electronic structure calculation, a model has been proposed to elucidate the conformation of copolymer chain in presence of various phthalates and their complexes, which offer the cause of higher gelation rate for longer aliphatic chain length up to n = 6, no gelation phenomena occurs for n > 6, and formation of two copolymer-solvent compounds. The mechanical properties (storage modulus and viscosity) decrease with increasing aliphatic chain length of phthalates and realignment of fibrils occurs at particular frequency depending on the strength of fibrillar gels. PMID- 19810754 TI - Structural basis for the lack of E2 interaction in the RING domain of TRAF2. AB - TRAF proteins are intracellular signal transducers for a number of immune receptor superfamilies. Specifically, TRAF2 interacts with members of the TNF receptor superfamily and connects the receptors to downstream signaling proteins. It has been assumed that TRAF2 is a ubiquitin ligase like TRAF6 and mediates K63 linked polyubiquitination of RIP1, a kinase pivotal in TNFalpha-induced NF-kappaB activation. Here we report the crystal structure of the RING and the first zinc finger domains of TRAF2. We show that the TRAF2 RING structure is very different from the known TRAF6 RING structure. The differences are multifaceted, including amino acid differences at the critical Ubc13-interacting site, local conformational differences, and a unique nine-residue insertion between the RING domain and the first zinc finger in TRAF2. These structural differences prevent TRAF2 from interacting with Ubc13 and other related E2s via steric clash and unfavorable interfaces. Our structural observation should prompt a re-evaluation of the role of TRAF2 in TNFalpha signaling and may indicate that TRAF2-associated proteins such as cIAPs may be the ubiquitin ligases for NF-kappaB signaling. PMID- 19810756 TI - Silicon oxide cluster formation and stability in the laser ablation of SiO targets. AB - The formation mechanism and stability of silicon oxide clusters observed in the ablation of SiO targets at 266 nm were investigated by time-of-flight mass spectrometry, laser-induced fluorescence (LIF), and DFT calculations. Neutral and positively charged Si(n)(+/0) and Si(n)O(m)H(0,1)(+) clusters were identified in the plume, but neutral Si(n)O(m) could not be observed. The time distribution of SiO in the plume measured by postionization with an ArF laser (Delta lambda approximately 1 nm, tau approximately 14 ns) and mass spectrometric detection was compared with that obtained by LIF with narrowband dye laser selective excitation of one specific rovibronic transition in SiO. Postionization leads to a multicomponent distribution that extends up to times near 100 micros after ablation, whereas LIF measurements obtain time distributions shorter than 20 micros. DFT calculations of several Si(n)O(m)(0/+) were performed, showing that one photon absorption of the postionization laser makes available low-energy dissociation channels of the neutrals, whereas two photon absorption is required for ionization. DFT calculations were carried out for stoichiometric H-containing clusters Si(n)O(n)H(+) (n = 1-4). For n = 1,2, the optimized geometries involve bonding of hydrogen to one oxygen atom in the clusters; for n = 3 and 4, the structures containing H-Si bonds are more stable. PMID- 19810757 TI - Computational investigation of the oxidative deboronation of boroglycine, H2N-CH2 B(OH)2, Using H2O and H2O2. AB - We report results from a computational investigation of the oxidative deboronation of boroglycine, H2N-CH2-B(OH)2, using H2O and H2O2 as the reactive oxygen species (ROS) to yield aminomethanol, H2N-CH2-OH; these results complement our study on the protodeboronation of boroglycine to produce methylamine, H2N-CH3 (Larkin et al. J. Phys. Chem. A 2007, 111, 6489-6500). Second-order Moller Plesset (MP2) perturbation theory with Dunning-Woon correlation-consistent (cc) basis sets were used for the calculations with comparisons made to results from density functional theory (DFT) at the PBE1PBE/6-311++G(d,p)(cc-pVDZ) levels. The effects of a bulk aqueous environment were also incorporated into the calculations employing PCM and CPCM methodology. Using H2O as the ROS, the reaction H2O + H2N-CH2-B(OH)2 --> H2N-CH2-OH + H-B(OH)2 was calculated to be endothermic; the value of DeltaH(298)(0) was +12.0 kcal/mol at the MP2(FC)/cc pVTZ computational level in vacuo and +13.7 kcal/mol in PCM aqueous media; the corresponding value for the activation barrier, DeltaH(double dagger), was +94.3 kcal/mol relative to the separated reactants in vacuo and +89.9 kcal/mol in PCM aqueous media. In contrast, the reaction H2O2 + H2N-CH2-B(OH)2 --> H2N-CH2-OH + B(OH)3 was calculated to be highly exothermic with an DeltaH(298)(0) value of 100.9 kcal/mol at the MP2(FC)/cc-pVTZ computational level in vacuo and -99.6 kcal/mol in CPCM aqueous media; the highest-energy transition state for the multistep process associated with this reaction involved the rearrangement of H2N CH2-B(OH)(OOH) to H2N-CH2-O-B(OH)2 with a DeltaH(double dagger) value of +23.2 kcal/mol in vacuo relative to the separated reactants. These computational results for boroglycine are in accord with the experimental observations for the deboronation of the FDA approved anticancer drug bortezomib (Velcade, PS-341), where it was found to be the principle deactivation pathway (Labutti et al. Chem. Res. Toxicol. 2006, 19, 539-546). PMID- 19810758 TI - Computational study on the stacking interaction in catechol complexes. AB - The stability and electron density topology of catechol complexes (dimers and tetramer) were studied using the MPW1B95 functional. The QTAIM analysis shows that both dimers (face to face and C-H/pi one) display a different electronic origin. The formation of the former is accompanied by a significant change in the values of atomic electron dipole and quadrupole components, flattening the most diffuse part of the electron density distribution toward the molecular plane. A small electron population transfer is observed between catechol monomers connected by C-H/pi interactions, whose QTAIM characterization does not differ from that of a weak hydrogen bond. Cooperative effects in the tetramer on binding energies are small and negligible for bond properties and charge transfer. Nevertheless, they are significant on atomic electron populations. PMID- 19810759 TI - Reactions of ionized methyl benzoate with methyl isocyanide in the gas phase: nucleophilic aromatic substitutions vs hydrogen migrations. AB - The chemistry leading to the competitive eliminations of H, CH(3), and OCOCH(3) from adducts of ionized methyl benzoate and neutral methyl isocyanide has been explored using density functional theory molecular orbital calculations. The energies of the various reactants and transition structures were estimated at the B3LYP/6-31+G(d,p) level of theory. Nucleophilic aromatic substitution is proposed to account for the H and OCOCH(3) eliminations. The corresponding sigma-complex intermediates, B(1ipso) and B(1ortho), are stable species lying in deep energy wells situated 70 and 120 kJ/mol, respectively, below the reactants, ionized methyl benzoate and methyl isocyanide. The latter complex, B(1ortho), may be also at the origin of a multistep rearrangement involving hydrogen migrations and methyl elimination from the original methoxy group of the benzoate moiety. PMID- 19810760 TI - Enantioselective copper-catalyzed reductive Michael cyclizations. AB - In the presence of siloxanes as stoichiometric reductants, chiral copper bisphosphine complexes catalyze highly enantioselective reductive Michael cyclizations of substrates containing two alpha,beta-unsaturated carbonyl moieties. The diastereochemical outcome of these reactions is dependent upon whether biaryl- or ferrocene-based chiral bisphosphines are employed. PMID- 19810761 TI - Catalytic enantioselective Cr-mediated propargylation: application to halichondrin synthesis. AB - A catalytic enantioselective propargylation in the presence of 10 mol % of Cr catalyst prepared from Cr(III) bromide and (R)-sulfonamide E furnishes homopropargyl alcohol 8 in 78% yield with 90% ee. Coupled with the workup based on Amano-lipase, this method provides a practical synthesis of optically pure 8 on a multigram scale. With maintenance of its optical purity, 8 has been converted to 1b, the C14-C19 building block of halichondrins and E7389, in two steps. PMID- 19810762 TI - Progress toward the total synthesis of (+/-)-actinophyllic acid. AB - This paper describes ongoing progress toward the synthesis of the novel indole alkaloid actinophyllic acid via a synthetic strategy that allows for the installation of all C-atoms (highlighted in red) requisite for completion of a total synthesis. PMID- 19810764 TI - Highly efficient oxidation of amines to imines by singlet oxygen and its application in Ugi-type reactions. AB - A variety of secondary benzylic amines were oxidized to imines in 90% to >99% yields by singlet oxygen generated from oxygen and a porphyrin photosensitizer. On the basis of these reactions, a protocol was developed for oxidative Ugi-type reactions with singlet oxygen as the oxidant. This protocol has been used to synthesize C1- and N-functionalized benzylic amines in up to 96% yields. PMID- 19810763 TI - Organocatalytic aziridine synthesis using F+ salts. AB - This paper describes a unique application of the fluoronium cation (F+) as an organocatalyst for mediating the reaction between N-substituted imines and ethyl diazoacetate affording excellent yields of N-substituted aziridines. PMID- 19810765 TI - Aliphatic imines in titanium-mediated reductive cross-coupling: unique reactivity of Ti(O-i-Pr)4/n-BuLi. AB - A procedure for the coupling of aliphatic imines with allylic and allenic alkoxides is described. The success of these studies was enabled by a unique reactivity profile of Ti(IV) isopropoxide/n-BuLi compared to well-known Ti(IV) isopropoxide/RMgX systems. PMID- 19810766 TI - Aerobic hydroxylation of N-borylenamine: triethylborane-mediated hydroxyalkylation of alpha,beta-unsaturated oxime ether. AB - Intermolecular hydroxyalkylation of alpha,beta-unsaturated imines involving Et3B mediated regioselective alkyl radical addition and subsequent hydroxylation with molecular oxygen has been developed, in which N-borylenamine generated by trapping of the enaminyl radical with Et3B was a key intermediate in the proposed aerobic hydroxylation mechanism. PMID- 19810767 TI - Silyl-substituted spirodiepoxides: stereoselective formation and regioselective opening. AB - A short synthesis of the natural product epi-citreodiol and the method developed to gain access to this target are described. Key advances focus on silyl substituted allenes. Upon exposure to dimethyldioxirane, spirodiepoxides form with high face selectivity and subsequently react at the silyl terminus. PMID- 19810768 TI - Catalytic enantioselective synthesis of chiral phthalides by efficient reductive cyclization of 2-acylarylcarboxylates under aqueous transfer hydrogenation conditions. AB - A new diamine ligand for asymmetric transfer hydrogenation (ATH) was discovered. The reductive cyclization of 2-acylarylcarboxylates was found to proceed highly stereoselectively by the new Ru complex-catalyzed ATH and subsequent in situ lactonization under aqueous conditions. It enables efficient access to a wide variety of 3-substituted phthalides in enantiomerically pure form. PMID- 19810769 TI - Pd-catalyzed C=C double-bond formation by coupling of N-tosylhydrazones with benzyl halides. AB - Pd-catalyzed reaction of N-tosylhydrazones with benzyl halides affords di- and trisubstituted olefins in high yields with excellent stereoselectivity. This coupling reaction is supposed to proceed through a migratory insertion of Pd carbene species. PMID- 19810770 TI - Stereoselective construction of substituted chromans by palladium-catalyzed cyclization of propargylic carbonates with 2-(2-hydroxyphenyl)acetates. AB - Highly substituted chromans have been constructed in a highly stereoselective manner by a palladium-catalyzed reaction of propargylic carbonates with 2-(2 hydroxyphenyl)acetates. Enantioselective reactions also successfully proceeded to give the optically active chromans with high enantioselectivity. PMID- 19810771 TI - Systems metabolic effects of a necator americanus infection in Syrian hamster. AB - Hookworms (Ancylostoma duodenale and Necator americanus) are blood-feeding intestinal nematodes that infect approximately 700 million people worldwide. To further our understanding of the systems metabolic response of the mammalian host to hookworm infection, we employed a metabolic profiling strategy involving the combination of (1)H NMR spectroscopic analysis of urine and serum and multivariate data analysis techniques to investigate the biochemical consequences of a N. americanus infection in the hamster. The infection was characterized by altered energy metabolism, consistent with hookworm-induced anemia. Additionally, disturbance of gut microbiotal activity was associated with a N. americanus infection, manifested in the alterations of microbial-mammalian cometabolites, including phenylacetylglycine, p-cresol glucuronide, 4-hydroxy-3-methyl phenylpropionic acid, hippurate, 4-hydroxyphenylactate, and dimethylamine. The correlation between worm burden and metabolite concentrations also reflected a changed energy metabolism and gut microbial state. Furthermore, elevated levels of urinary 2-aminoadipate was a characteristic feature of the infection, which may be associated with the documented neurological consequences of hookworm infection. PMID- 19810773 TI - Do biological medicinal products pose a risk to the environment?: a current view on ecopharmacovigilance. AB - The occurrence of active pharmaceutical substances in the environment is of growing concern. The vast majority of the compounds in question are of low molecular weight, intended for oral use and designed to tolerate, for example, the digestive enzymes in the upper alimentary tract, the harsh milieus found in the acidic stomach, or the microbe rich intestine. Accordingly, these xenobiotic compounds may, due to their inherent biological activity, constitute a risk to the environment. Biological medicinal products, for example recombinant human insulin or monoclonal antibodies, however, are different. They are primarily made up of oligomers or polymers of amino acids, sugars or nucleotides and are thus readily metabolized. They are therefore generally not considered to pose any risk to the environment. Certain classes of biological medicinal products, however, are associated with specific safety issues. Genetically modified organisms as vectors in vaccines or in gene therapy products have attracted much attention in this regard. Issues include the degree of attenuation of the live recombinant vaccine, replication restrictions of the vaccine vector, alteration of the host and tissue tropism of the vector, the possibility of reversion to virulence, and risk to the ecosystem. In this review we discuss the fate and the potential environmental impact of biological medicinal products following clinical use from an ecopharmacovigilance point of view, and review relevant policy documents and regulatory statements. PMID- 19810772 TI - Annexin A5 directly interacts with amyloidogenic proteins and reduces their toxicity. AB - Protein misfolding is a central mechanism for the development of neurodegenerative diseases and type 2 diabetes mellitus. The accumulation of misfolded alpha-synuclein protein inclusions in the Lewy bodies of Parkinson's disease is thought to play a key role in pathogenesis and disease progression. Similarly, the misfolding of the beta-cell hormone human islet amyloid polypeptide (h-IAPP) into toxic oligomers plays a central role in the induction of beta-cell apoptosis in the context of type 2 diabetes. In this study, we show that annexin A5 plays a role in interacting with and reducing the toxicity of the amyloidogenic proteins, h-IAPP and alpha-synuclein. We find that annexin A5 is coexpressed in human beta-cells and that exogenous annexin A5 reduces the level of h-IAPP-induced apoptosis in human islets by approximately 50% and in rodent beta-cells by approximately 90%. Experiments with transgenic expression of alpha synuclein in Caenorhabditis elegans show that annexin A5 reduces alpha-synuclein inclusions in vivo. Using thioflavin T fluorescence, electron microscopy, and electron paramagnetic resonance, we provide evidence that substoichiometric amounts of annexin A5 inhibit h-IAPP and alpha-synuclein misfolding and fibril formation. We conclude that annexin A5 might act as a molecular safeguard against the formation of toxic amyloid aggregates. PMID- 19810774 TI - A benefit-risk assessment of imatinib in chronic myeloid leukaemia and gastrointestinal stromal tumours. AB - Targeting constitutively activated tyrosine kinases, such as BCR-ABL, in chronic myeloid leukaemia (CML) and c-KIT in gastrointestinal stromal tumours (GIST) has substantially changed the clinical management of both diseases. The introduction of imatinib, a tyrosine kinase inhibitor mainly targeting BCR-ABL, c-KIT and PDGFR, has profoundly improved the prognosis of both entities, while being surprisingly well tolerated. This article summarizes recent data on clinical efficacy as well as safety aspects of imatinib for treatment of CML and GIST, including a final benefit-risk assessment. Imatinib induces high rates of cytogenetic and molecular responses in all phases of CML and also has substantial activity in GIST patients. In both diseases, only a few adverse effects, such as musculoskeletal and joint pain, muscle cramps, oedema and gastrointestinal symptoms, occur. Most of these are grade I or II toxicities and generally occur during the early phase of treatment (i.e. within the first 2 years). Thus, in view of the low rates of severe toxicities and the extraordinary efficacy of the drug in both diseases, imatinib represents an oral drug with a high benefit-risk ratio for the treatment of CML and GIST. PMID- 19810775 TI - Predicting the clinical relevance of drug interactions from pre-approval studies. AB - Drug interactions (DIs) may result in adverse drug events that could be prevented, but in many cases the available information on potential DIs is not easily transferable to clinical practice. The majority of studies date from preclinical or premarketing phases, using animals or human-derived sources that may not accurately reflect the growing clinical complexity of high-risk populations, such as the elderly, women, children, patients with chronic disease, polytherapy and impaired organ functions. Thus, at the time of approval of a new drug the information in the summary of product characteristics refers to potential DIs, but lacks specific management recommendations and is of limited clinical utility. Therefore, we set out to review in vitro and in vivo methods to predict and quantify potential DIs, to see whether these studies could help the physician tackle daily problems of the assessment and choice of combined drug therapies, and to propose, from a clinical point of view, how premarketing studies could be improved so as to help the physician at the patient's bedside. Preclinical and premarketing study design needs to be improved to make information easily accessible and clinically transferable. Studies should also take into account appropriate sample size, duration, co-morbidity, number of coadministered drugs, within- and between-subject variability, specific at-risk populations and/or drugs with a relatively narrow therapeutic window, and clinical endpoints. After premarketing development in situations where there is potential high risk of serious adverse events, specific phase IV studies (and/or active pharmacovigilance studies) should be required to monitor and quantitatively assess their clinical impact. PMID- 19810778 TI - Consumer reporting of adverse drug reactions: a retrospective analysis of the Danish adverse drug reaction database from 2004 to 2006. AB - BACKGROUND: Reporting adverse drug reactions (ADRs) has traditionally been the sole province of healthcare professionals. Since 2003 in Denmark, consumers have been able to report ADRs directly to the authorities. The objective of this study was to compare ADRs reported by consumers with ADRs reported from other sources, in terms of their type, seriousness and the suspected medicines involved. METHODS: The number of ADRs reported to the Danish ADR database from 2004 to 2006 was analysed in terms of category of reporter, seriousness, category of ADRs by system organ class (SOC) and the suspected medicines on level 1 of the anatomical therapeutic chemical (ATC) classification system. ADR reports from consumers were compared with reports from other sources (physicians, pharmacists, lawyers, pharmaceutical companies and other healthcare professionals). Chi-square and odds ratios (ORs) were calculated to investigate the dependence between type of reporter and reported ADRs (classified by ATC or SOC). FINDINGS: We analysed 6319 ADR reports corresponding to 15 531 ADRs. Consumers reported 11% of the ADRs. Consumers' share of 'serious' ADRs was comparable to that of physicians (approximately 45%) but lower than that of pharmacists and other healthcare professionals. When consumer reports were compared with reports from other sources, consumers were more likely to report ADRs from the following SOCs: 'nervous system disorders' (OR = 1.27; 95% CI 1.05, 1.53); 'psychiatric disorders' (OR = 1.70; 95% CI 1.31, 2.20) and 'reproductive system and breast disorders' (OR = 2.02; 95% CI 1.13, 3.61) than other sources. Compared with other sources, consumers reported fewer ADRs from the SOCs 'blood and lymphatic system disorders' (OR = 0.22; 95% CI 0.08, 0.59) and 'hepatobiliary system disorders' (OR = 0.14; 95% CI 0.04, 0.57). Consumers were more likely to report ADRs from the ATC group N (nervous system) [OR = 2.72; 95% CI 2.34, 3.17], ATC group P (antiparasitic products) [OR = 2.41; 95% CI 1.32, 4.52] and ATC group S (sensory organs) [OR = 4.79; 95% CI 2.04, 11.23] than other sources. Consumers reported fewer ADRs from the ATC group B (blood and blood-forming organs) [OR = 0.04; 95% CI 0.006, 0.32] and the ATC groups J (anti-infective for systemic use) [OR = 0.44; 95% CI 0.33, 0.58], L (antioneoplastic and immunomodulating agents) [OR = 0.19; 95% CI 0.12, 0.30] and V (various) [OR = 0.03; 95% CI 0.004, 0.21] than other sources. In the SOC 'nervous system disorders', consumers reported seven categories of ADRs that were not reported by the other sources. CONCLUSION: This study showed that compared with other sources, consumers reported different categories of ADRs for different types of medicines. Consumers should be actively included in systematic drug surveillance systems, including clinical settings, and their reports should be taken as seriously as reports from other sources. PMID- 19810777 TI - Risk management policy and black-box warnings: a qualitative analysis of US FDA proceedings. AB - BACKGROUND: The US FDA increasingly applies risk management to drug safety policy. Little is known about the process by which the FDA approves labelling changes. Although advisory committees can recommend any of the risk management tools, including the use of 'black-box warnings', it is unknown whether they deliberate on these questions or how they apply the principles of risk minimization or management during their considerations of drug licensing. OBJECTIVE: To examine the process by which risk management is considered by the FDA, including the role of FDA advisory committees. We also aimed to identify and describe drug labelling changes and additions, including the prevalence of black box warnings. METHODS: We electronically obtained publicly available information regarding drug approvals, drug revisions and advisory committee meetings over 3 years (2004-6) from the FDA. Data in the form of meeting transcripts and full histories of labelling changes were collected on drugs discussed by advisory committees. We then searched and qualitatively analysed the meeting transcripts to identify themes in the discussion. We also created a database of all prescription drug labelling changes for 3 years and examined which drugs have had the most changes. We describe the risk management consideration process and report the frequency and characteristics of labelling changes. Excerpts from the transcripts are selected to illustrate both typical and atypical features of the discussion. FINDINGS: A total of 174 black-box changes were made in the 3-year period of our study, of which 77 were new black-box warnings and 97 were revisions in black-box warnings. Of 77 new black-box warning additions, only 11 drugs were discussed by the advisory committees. Of the 17 most frequently revised drug labels in these 3 years, two were discussed in the advisory committee meetings. Advisory meeting discussions revealed confusion about black box warnings and emphasized potential consequences of the warnings rather than their content. CONCLUSION: The safety labelling of drugs on the market is changed often. Panels of advisors consider only a few drugs, rarely discuss the labelling requirements, and display confusion about applying black-box warnings. The creation and application of black-box warnings on prescription medications should receive closer attention from the FDA and its advisors. PMID- 19810776 TI - Differences in adverse effect reporting in placebo groups in SSRI and tricyclic antidepressant trials: a systematic review and meta-analysis. AB - BACKGROUND: Biases in adverse effect reporting in randomized controlled trials (RCTs) [e.g. due to investigator expectations or assessment quality] can be quantified by studying the rates of adverse events reported in the placebo arms of such trials. OBJECTIVE: We compared the rates of adverse effects reported in the placebo arms of tricyclic antidepressant (TCA) trials and placebo arms of selective serotonin reuptake inhibitor (SSRI) trials. METHODS: We conducted a literature search for RCTs across PUBMED, Scopus and the Cochrane Central Register of Controlled Trials (CENTRAL). Only studies allowing adverse effect analysis were included. Publication year ranged from 1981 to 2007. RESULTS: Our systematic review and meta-analysis included 143 placebocontrolled RCTs and data from 12,742 patients. Only 21% of studies used structured and systematic adverse effect ascertainment strategies. The way in which trials recorded adverse events influenced the rate of adverse effects substantially. Systematic assessment led to higher rates than less systematic assessment. Far more adverse effects were reported in TCA-placebo groups compared with SSRI-placebo groups, e.g. dry mouth (odds ratio [OR] = 3.5; 95% CI 2.9, 4.2); drowsiness (OR = 2.7; 95%CI 2.2, 3.4); constipation (OR= 2.7; 95%CI 2.1, 3.6); sexual problems (OR =2.3; 95%CI 1.5, 3.5). Regression analyses controlling for various influencing factors confirmed the results. CONCLUSION: Adverse effect profiles reported in clinical trials are strongly influenced by expectations from investigators and patients. This difference cannot be attributed to ascertainment methods. Adverse effect patterns of the drug group are closely related to adverse effects of the placebo group. These results question the validity of the assumption that adverse effects in placebo groups reflect the 'drug-unspecific effects'. PMID- 19810779 TI - Effectiveness of safety warnings in atypical antipsychotic drugs: an interrupted time-series analysis in Spain. AB - BACKGROUND: Studies conducted to obtain drug authorization are often of short duration and based on small sample sizes in selected populations. Policies on drug safety rely on the validity of the methods used to achieve rapid and effective communication of new information. No formal evaluation has ever been made of the Spanish communications system, although indirect data have raised questions about its effectiveness. OBJECTIVE: To evaluate the impact of two safety warnings issued by the Spanish Drug Agency, and of a later prior authorization requirement involving the use of atypical antipsychotic drugs in the elderly. METHODS: The study was based on a time-series analysis constructed with data corresponding to monthly invoicing from 2000 to 2006 for olanzapine and risperidone in the Region of Valencia, Spain. Because the safety warnings and the prior authorization policy applied exclusively to prescriptions of these drugs for elderly patients with dementia, we investigated whether these interventions were successful and therefore changed prescription patterns for pensioners receiving low-strength formulations (the available proxy for elderly subjects with dementia), without altering patterns for those receiving the highest strength formulations (typically used in schizophrenic patients) or for prescriptions for non-pensioners (any strength formulations). These two latter groups were therefore established as the control groups. RESULTS: Defined daily doses (DDDs) for olanzapine in low-strength pharmaceutical forms showed a clear levelling off after the first warning, while that for risperidone showed less pronounced decline. The prior authorization policy had a dramatic effect on the consumption of risperidone, but not on that of olanzapine. DDDs for low-strength formulations between the 12 months prior to the first warning and the 12 months following the prior authorization showed a substantial reduction (22% for risperidone and 33% for olanzapine). In the high-strength forms and in non pensioners the upward trends in DDDs remained unaltered after both interventions. CONCLUSION: The safety warnings concerning atypical antipsychotic drugs were effective in reducing the prescribing of risperidone and olanzapine in low strength doses in pensioner prescriptions, and the implementation of a prior authorization policy had a dramatic effect on the prescribing of risperidone. PMID- 19810780 TI - Mortality associated with attention-deficit hyperactivity disorder (ADHD) drug treatment: a retrospective cohort study of children, adolescents and young adults using the general practice research database. AB - BACKGROUND: Following reports of sudden death in patients taking medication to treat attention-deficit hyperactivity disorder (ADHD), this study aimed to identify cases of death in patients prescribed stimulants and atomoxetine and to determine any association between these and sudden death. METHOD: The UK General Practice Research Database (GPRD) was used to identify patients aged 2-21 years from 1 January 1993 to 30 June 2006 with a prescription for methylphenidate, dexamfetamine or atomoxetine. Patients were followed from the date of first prescription until whichever came first: date of death, transferred-out date, age >21 years or end of the study period. RESULTS: From a cohort of 18 637 patient years, seven patients died. Cause of death was obtained for six of the patients. None were deemed to be cases of sudden death. Compared with a general population rate of sudden death in the literature, the worst-case scenario of one case of sudden death gave an incident rate ratio of 1.63 (95% CI 0.04, 9.71). Although it was not the primary outcome of the study, following reports of suicide in the cohort the standardized mortality ratio (SMR) of suicide was calculated. Due to differences in the definition of suicide used for children and adults, patients were categorized into two age groups: 11-14 years and 15-21 years. The SMR for suicide for patients aged 11-14 years was 161.91 (95% CI 19.61, 584.88). The SMR for suicide for patients aged 15-21 years was 1.84 (95% CI 0.05, 10.25). CONCLUSION: This study demonstrated no increase in the risk of sudden death associated with stimulants or atomoxetine. However, an increased risk of suicide was seen. Although we cannot exclude that the medications may contribute to the increased risk of suicide, there are other factors such as depression and antisocial behaviour that frequently co-exist with ADHD, which can also predispose to teenage suicide. Clinicians should identify patients at increased risk of cardiovascular events and identify those patients at increased risk of suicide, particularly males with co-morbid conditions, and monitor them appropriately. PMID- 19810781 TI - Attention-deficit hyperactivity disorder (ADHD) treatment and sudden death. PMID- 19810783 TI - Accumulation of visual information across multiple fixations. AB - Humans often redirect their gaze to the same objects within a scene, even without being consciously aware of it. Here, we investigated what type of visual information is accumulated across recurrent fixations on the same object. On each trial, subjects viewed an array comprised of several objects and were subsequently asked to report on various visual aspects of a randomly chosen target object from that array. Memory performance decreased as more fixations were directed to other objects, following the last fixation on the target object (i.e. post-target fixations). In contrast, performance was enhanced with increasing number of fixations on the target object. However, since the number of post-target fixations and the number of target fixations are usually anti correlated, memory gain may simply reflect fewer post-target fixations, rather than true accumulation of information. To rule this out, we conducted a second experiment, in which the stimulus disappeared immediately after performing a predefined number of target fixations. Additional fixations on the target object resulted in improved memory performance even under these strict conditions. We conclude that, under the present conditions, various aspects of memory monotonically improve with repeated sampling of the same object. PMID- 19810782 TI - Contrast sensitivity in natural scenes depends on edge as well as spatial frequency structure. AB - The contrast sensitivity function is routinely measured in the laboratory with sine-wave gratings presented on homogenous gray backgrounds; natural images are instead composed of a broad range of spatial and temporal structures. In order to extend channel-based models of visual processing to more natural conditions, we examined how contrast sensitivity varies with the context in which it is measured. We report that contrast sensitivity is quite different under laboratory than natural viewing conditions: adaptation or masking with natural scenes attenuates contrast sensitivity at low spatial and temporal frequencies. Expressed another way, viewing stimuli presented on homogenous screens overcomes chronic adaptation to the natural environment and causes a sharp, unnatural increase in sensitivity to low spatial and temporal frequencies. Consequently, the standard contrast sensitivity function is a poor indicator of sensitivity to structure in natural scenes. The magnitude of masking by natural scenes is relatively independent of local contrast but depends strongly on the density of edges even though neither greatly affects the local amplitude spectrum. These results suggest that sensitivity to spatial structure in natural scenes depends on the distribution of local edges as well as the local amplitude spectrum. PMID- 19810784 TI - From disparity to depth: how to make a grating and a plaid appear in the same depth plane. AB - Even though binocular disparity is a very well-studied cue to depth, the function relating disparity and perceived depth has been characterized only for the case of horizontal disparities. We sought to determine the general relationship between disparity and depth for a particular set of stimuli. The horizontal disparity direction is a special case, albeit an especially important one. Non horizontal disparities arise from a number of sources under natural viewing condition. Moreover, they are implicit in patterns that are one-dimensional, such as gratings, lines, and edges, and in one-dimensional components of two dimensional patterns, where a stereo matching direction is not well-defined. What function describes perceived depth in these cases? To find out, we measured the phase disparities that produced depth matches between a reference stimulus and a test stimulus. The reference stimulus was two-dimensional, a plaid; the test stimulus was one-dimensional, a grating. We find that horizontal disparity is no more important than other disparity directions in determining depth matches between these two stimuli. As a result, a grating and a plaid appear equal in depth when their horizontal disparities are, in general, unequal. Depth matches are well predicted by a simple disparity vector calculation; they survive changes in component parameters that conserve these vector quantities. The disparity vector rule also describes how the disparities of 1-D components might contribute to the perceived depth of 2-D stimuli. PMID- 19810785 TI - Recovery of a crowded object by masking the flankers: determining the locus of feature integration. AB - Object recognition is a central function of the visual system. As a first step, the features of an object are registered; these independently encoded features are then bound together to form a single representation. Here we investigate the locus of this "feature integration" by examining crowding, a striking breakdown of this process. Crowding, an inability to identify a peripheral target surrounded by flankers, results from "excessive integration" of target and flanker features. We presented a standard crowding display with a target C flanked by four flanker C's in the periphery. We then masked only the flankers (but not the target) with one of three kinds of masks-noise, metacontrast, and object substitution-each of which interferes at progressively higher levels of visual processing. With noise and metacontrast masks (low-level masking), the crowded target was recovered, whereas with object substitution masks (high-level masking), it was not. This places a clear upper bound on the locus of interference in crowding suggesting that crowding is not a low-level phenomenon. We conclude that feature integration, which underlies crowding, occurs prior to the locus of object substitution masking. Further, our results indicate that the integrity of the flankers, but not their identification, is crucial for crowding to occur. PMID- 19810786 TI - The relationship between object spatial profile and accommodation microfluctuations in emmetropes and myopes. AB - The accommodation microfluctuations are thought to be used by the accommodation controller to obtain information about the direction and magnitude of the required response by monitoring changes in the contrast gradient of this image. The contrast gradient can be altered by presenting different spatial frequency (SF) targets to the eye. Twelve myopes (MYOs) and 12 emmetropes (EMMs) viewed sine and square wave targets of SF 0.5, 1, 2, 4, 8, 16 cpd in a Badal optical system. Accommodation responses were recorded continuously using the Shin-Nippon SRW-5000 autorefractor. There is no change in magnitude of the accommodation microfluctuations as the SF of square waves is altered. While viewing sine wave targets, the microfluctuations are smallest for mid (2, 4 cpd) SFs and increase for low (0.5 cpd) and high (16 cpd) SFs. MYOs show a significantly larger increase in the microfluctuations for the 16 cpd target compared to the EMMs. MYOs have significantly larger microfluctuations than the EMMs throughout. The microfluctuations seem to be monitoring the contrast gradient of the cortical image, which is likely to be used by the accommodation control system during error detection. The results indicate that MYO subjects may have a shallower contrast gradient and the potential reasons and implications of this are discussed. PMID- 19810787 TI - The contributions of central versus peripheral vision to scene gist recognition. AB - Which region of the visual field is most useful for recognizing scene gist, central vision (the fovea and parafovea) based on its higher visual resolution and importance for object recognition, or the periphery, based on resolving lower spatial frequencies useful for scene gist recognition, and its large extent? Scenes were presented in two experimental conditions: a "Window," a circular region showing the central portion of a scene, and blocking peripheral information, or a "Scotoma," which blocks out the central portion of a scene and shows only the periphery. Results indicated the periphery was more useful than central vision for maximal performance (i.e., equal to seeing the entire image). Nevertheless, central vision was more efficient for scene gist recognition than the periphery on a per-pixel basis. A critical radius of 7.4 degrees was found where the Window and Scotoma performance curves crossed, producing equal performance. This value was compared to predicted critical radii from cortical magnification functions on the assumption that equal V1 activation would produce equal performance. However, these predictions were systematically smaller than the empirical critical radius, suggesting that the utility of central vision for gist recognition is less than predicted by V1 cortical magnification. PMID- 19810788 TI - The precision of visual working memory is set by allocation of a shared resource. AB - The mechanisms underlying visual working memory have recently become controversial. One account proposes a small number of memory "slots," each capable of storing a single visual object with fixed precision. A contrary view holds that working memory is a shared resource, with no upper limit on the number of items stored; instead, the more items that are held in memory, the less precisely each can be recalled. Recent findings from a color report task have been taken as crucial new evidence in favor of the slot model. However, while this task has previously been thought of as a simple test of memory for color, here we show that performance also critically depends on memory for location. When errors in memory are considered for both color and location, performance on this task is in fact well explained by the resource model. These results demonstrate that visual working memory consists of a common resource distributed dynamically across the visual scene, with no need to invoke an upper limit on the number of objects represented. PMID- 19810789 TI - Globally inconsistent figure/ground relations induced by a negative part. AB - Figure/ground interpretation is a dynamic and complex process involving the cooperation and competition of a number of perceptual factors. Most research has assumed that figure/ground assignment is globally consistent along the entire contour of a single figure, meaning that the one side of each boundary is interpreted as figure along the entire length of the boundary, and the other side interpreted as ground. We investigated a situation that challenges this assumption, because local cues to figure/ground conflict with global cues: a "negative part," a contour region that appears locally convex but that the global form requires be concave. To measure figure/ground assignment, we use a new task based on local contour motion attribution that allows us to measure border ownership locally at points along the contour. The results from two experiments showed that the more salient a negative part is, the more border ownership tended to locally reverse within it, creating an inconsistency in figure/ground assignments along the contour. This suggests that border ownership assignment is not an all-or-none process, but rather a locally autonomous process that is not strictly constrained by global cues. PMID- 19810790 TI - Recruitment of a novel cue for active control depends on control dynamics. AB - We investigated how the visual-motor system recruits a novel visual feedback cue for a manual control task. We presented conditions in which an arbitrary cue (color) was coupled with task-relevant feedback (position or velocity), and measured the effect of the novel cue on performance. Participants used a joystick to keep a moving horizontal line centered on a display under velocity or acceleration control dynamics. Participants normally rely primarily on line position feedback for velocity control and line velocity feedback for acceleration control. The novel color cue was coupled with either line position (becoming red as it deviates from center) or line velocity (becoming red as it moves faster). For velocity control, performance error was smaller and response gain was larger when the novel color cue was coupled with line position than when it was coupled with line velocity. Conversely, for acceleration control, performance was better when color was coupled with line velocity than with line position. Our findings show that the visual-motor system can recruit a novel arbitrary cue to improve active control performance, but the effectiveness of the novel cue depends on its relationship to the feedback appropriate for control dynamics. PMID- 19810792 TI - Perception of limited-lifetime biological motion from different viewpoints. AB - Studies with time-limited point-lights suggested that biological motion does not require local motion detection. These studies used walkers seen from the side, but biological motion perception excels also when walkers are oriented toward the observer, or in intermediate, half-profile views. In perspective projection, the local motion of points on the body provides a cue to the 3D structure of the walker. Thus, local point motion that was irrelevant for walkers in profile view may become important for biological motion perception in perspective projection. We compared performance on forward/backward walking discrimination of walkers in orthographic and perspective projection when view orientations and with point lifetime was varied. We found no difference between orthographic and perspective projections. Walkers with point lifetime 1 allowed forward-backward discrimination reliably in non-profile views, suggesting that local image motion is not required. Discrimination performance became extremely difficult in the frontal view, however. Follow-up experiments that tested lifetime, view orientation, and specific information from the feet indicated that this dependence on viewing angle can be explained by the reliance of the forward/backward discrimination on information about the movement of the lower legs, which is difficult to ascertain in the frontal view. PMID- 19810791 TI - The gender-specific face aftereffect is based in retinotopic not spatiotopic coordinates across several natural image transformations. AB - In four experiments, we measured the gender-specific face-aftereffect following subject's eye movement, head rotation, or head movement toward the display and following movement of the adapting stimulus itself to a new test location. In all experiments, the face aftereffect was strongest at the retinal position, orientation, and size of the adaptor. There was no advantage for the spatiotopic location in any experiment nor was there an advantage for the location newly occupied by the adapting face after it moved in the final experiment. Nevertheless, the aftereffect showed a broad gradient of transfer across location, orientation and size that, although centered on the retinotopic values of the adapting stimulus, covered ranges far exceeding the tuning bandwidths of neurons in early visual cortices. These results are consistent with a high-level site of adaptation (e.g. FFA) where units of face analysis have modest coverage of visual field, centered in retinotopic coordinates, but relatively broad tolerance for variations in size and orientation. PMID- 19810793 TI - Temporal whitening: transient noise perceptually equalizes the 1/f temporal amplitude spectrum. AB - Naturally occurring luminance distributions are approximately 1/f in their spatial and temporal amplitude spectra. By systematically varying the spatio temporal profile of broadband noise stimuli, we demonstrate that humans invariably overestimate the proportion of high spatial and temporal frequency energy. Critically, we find that that the strength of this bias is of a magnitude that predicts a perceptually equalized response to the spatio-temporal fall off in the natural amplitude spectrum. This interpretation is supported by our finding that the magnitude of this transient response bias, while evident across a broad range of narrowband spatial frequencies (0.25-8 cycles/deg), decreases above 2 cycles/deg, which itself compensates for the increase in temporal frequency energy previously observed at high spatial frequencies as a consequence of small fixational eye movements (M. Rucci, R. Iovin, M. Poletti, & F. Santini, 2007). Additional temporal masking and adaptation experiments reveal a transiently biased asymmetry. Whereas temporal frequencies >4 Hz mask and adapt 1 and 15-Hz targets, lower masking and adaptation frequencies have much less effect on sensitivity to 15-Hz compared with 1-Hz targets. These results imply that the visual system over-represents its transient input to an extent that predicts an equalized temporal channel response to the low-frequency-biased structure of natural scenes. PMID- 19810794 TI - The aperture problem in contoured stimuli. AB - A moving object elicits responses from V1 neurons tuned to a broad range of locations, directions, and spatiotemporal frequencies. Global pooling of such signals can overcome their intrinsic ambiguity in relation to the object's direction/speed (the "aperture problem"); here we examine the role of low-spatial frequencies (SF) and second-order statistics in this process. Subjects made a 2AFC fine direction-discrimination judgement of 'naturally' contoured stimuli viewed rigidly translating behind a series of small circular apertures. This configuration allowed us to manipulate the scene by randomly switching which portion of the stimulus was presented behind each aperture or by occluding certain spatial frequency bands. We report that global motion integration is (a) largely insensitive to the second-order statistics of such stimuli and (b) is rigidly broadband even in the presence of a disrupted low SF component. PMID- 19810795 TI - The role of orientation and position in shape perception. AB - This study investigates the contributions of position versus orientation information in shape perception by putting the two in conflict. Sampling the orientation of, e.g., a rounded pentagon and positioning the samples on a circle creates a stimulus in which element positions are consistent with a circle but element orientations with a pentagon. Whether orientation or position dominates the percept depends on a number of factors. First, perceived shape shows a band pass relationship with respect to number of samples. Element orientation captures element position unless elements are widely separated or very closely spaced. This effect is scale invariant. Second, increasing element envelope size or decreasing carrier wavelength strengthens the influence of element orientation, while other parameters such as the phase and polarity of the carrier or the scale of the Gabor are irrelevant. Third, the overall shape of the contour modulates the effect. The strength of the positional signal rises as the orientation difference between adjacent elements increases. Consequently, the computation underlying contour shape relies on a weighted combination of element orientation and position with weights, not fixed, but dependent on stimulus details. When orientation is dominant, its signal is strong enough to alter positional information, giving rise to the illusion of, e.g., a pentagon despite elements being on a circle. PMID- 19810796 TI - Myopia and peripheral ocular aberrations. AB - We measured wave aberrations over the central 42 degrees x 32 degrees visual field for a 5-mm pupil for groups of 10 emmetropic (mean spherical equivalent = 0.11 +/- 0.50 D) and 9 myopic (MSE = -3.67 +/- 1.91 D) young adults. Relative peripheral refractive errors over the measured field were generally myopic in both groups. Mean values of C(4)(0) were almost constant across the measured field and were more positive in emmetropes (+0.023 +/- 0.043 microm) than in myopes (-0.007 +/- 0.045 microm). Coma varied more rapidly with field angle in myopes: modeling suggested that this difference reflected the differences in mean anterior corneal shape and axial length in the two groups. In general, however, overall levels of RMS aberration differed only modestly between the two groups, implying that it is unlikely that high levels of aberration contribute to myopia development. PMID- 19810797 TI - Intermittent occlusion enhances the smoothness of sampled motion. AB - Discrete sequences of sampled motion often appear flickering and jerky. We present evidence showing that a target in sampled motion is perceived as smoother when structure in the background appears and disappears synchronously with the target. Specifically, we found that target flicker is turned into permanent target visibility at short interstimulus intervals and jerkiness is replaced by smoothly accelerating and decelerating motion at longer ones. We argue that this "smoothening" effect is essentially a form of amodal completion in space-time being evoked by spatiotemporal cues to occlusion. The effect highlights the importance of amodal representations in perception. PMID- 19810798 TI - Improved classification images with sparse priors in a smooth basis. AB - Classification images provide compelling insight into the strategies used by observers in psychophysical tasks. However, because of the high-dimensional nature of classification images and the limited quantity of trials that can practically be performed, classification images are often too noisy to be useful unless denoising strategies are adopted. Here we propose a method of estimating classification images by the use of sparse priors in smooth bases and generalized linear models (GLMs). Sparse priors in a smooth basis are used to impose assumptions about the simplicity of observers' internal templates, and they naturally generalize commonly used methods such as smoothing and thresholding. The use of GLMs in this context provides a number of advantages over classic estimation techniques, including the possibility of using stimuli with non Gaussian statistics, such as natural textures. Using simulations, we show that our method recovers classification images that are typically less noisy and more accurate for a smaller number of trials than previously published techniques. Finally, we have verified the efficiency and accuracy of our approach with psychophysical data from a human observer. PMID- 19810799 TI - Sensitivity and perceptual awareness increase with practice in metacontrast masking. AB - Can practice effects on unconscious stimuli lead to awareness? Can we "learn to see"? Recent evidence suggests that blindsight patients trained for an extensive period of time can learn to discriminate and consciously perceive stimuli that they were previously unaware of. So far, it is unknown whether these effects generalize to normal observers. Here we investigated practice effects in metacontrast masking. Subjects were trained for five consecutive days on the stimulus onset asynchrony (SOA) that resulted in chance performance. Our results show a linear increase in sensitivity (d') but no change in bias (c) for the trained SOA. This practice effect on sensitivity spreads to all tested SOAs. Additionally, we show that subjects rate their perceptual awareness of the target stimuli differently before and after training, exhibiting not only an increase in sensitivity, but also in the subjective awareness of the percept. Thus, subjects can indeed "learn to see." PMID- 19810801 TI - Occlusion-related lateral connections stabilize kinetic depth stimuli through perceptual coupling. AB - Local sensory information is often ambiguous forcing the brain to integrate spatiotemporally separated information for stable conscious perception. Lateral connections between clusters of similarly tuned neurons in the visual cortex are a potential neural substrate for the coupling of spatially separated visual information. Ecological optics suggests that perceptual coupling of visual information is particularly beneficial in occlusion situations. Here we present a novel neural network model and a series of human psychophysical experiments that can together explain the perceptual coupling of kinetic depth stimuli with activity-driven lateral information sharing in the far depth plane. Our most striking finding is the perceptual coupling of an ambiguous kinetic depth cylinder with a coaxially presented and disparity defined cylinder backside, while a similar frontside fails to evoke coupling. Altogether, our findings are consistent with the idea that clusters of similarly tuned far depth neurons share spatially separated motion information in order to resolve local perceptual ambiguities. The classification of far depth in the facilitation mechanism results from a combination of absolute and relative depth that suggests a functional role of these lateral connections in the perception of partially occluded objects. PMID- 19810802 TI - A comprehensive transcriptomic analysis of the effect of silicon on wheat plants under control and pathogen stress conditions. AB - The supply of soluble silicon (Si) to plants has been associated with many benefits that remain poorly explained and often contested. In this work, the effect of Si was studied on wheat plants under both control and pathogen stress (Blumeria graminis f. sp. tritici) conditions by conducting a large transcriptomic analysis (55,000 unigenes) aimed at comparing the differential response of plants under four treatments. The response to the supply of Si on control (uninfected) plants was limited to 47 genes of diverse functions providing little evidence of regulation of a specific metabolic process. Plants reacted to inoculation with B. graminis f. sp. tritici by an upregulation of many genes linked to stress and metabolic processes and a downregulation of genes linked to photosynthesis. Supplying Si to inoculated plants largely prevented disease development, a phenotypic response that translated into a nearly perfect reversal of genes regulated by the effect of B. graminis f. sp. tritici alone. These results suggest that Si plays a limited role on a plant's transcriptome in the absence of stress, even in the case of a high-Si-accumulating monocot such as wheat. On the other hand, the benefits of Si in the form of biotic stress alleviation were remarkably aligned with a counter-response to transcriptomic changes induced by the pathogen B. graminis f. sp. tritici. PMID- 19810800 TI - Effects of gestational length, gender, postnatal age, and birth order on visual contrast sensitivity in infants. AB - To investigate effects of visual experience versus preprogrammed mechanisms on visual development, we used multiple regression analysis to determine the extent to which a variety of variables (that differ in the extent to which they are tied to visual experience) predict luminance and chromatic (red/green) contrast sensitivity (CS), which are mediated by the magnocellular (M) and parvocellular (P) subcortical pathways, respectively. Our variables included gestational length (GL), birth weight (BW), gender, postnatal age (PNA), and birth order (BO). Two month-olds (n = 60) and 6-month-olds (n = 122) were tested. Results revealed that (1) at 2 months, infants with longer GL have higher luminance CS; (2) at both ages, CS significantly increases over a approximately 21-day range of PNA, but this effect is stronger in 2- than 6-month-olds and stronger for chromatic than luminance CS; (3) at 2 months, boys have higher luminance CS than girls; and (4) at 2 months, firstborn infants have higher CS, while at 6 months, non-firstborn infants have higher CS. The results for PNA/GL are consistent with the possibility that P pathway development is more influenced by variables tied to visual experience (PNA), while M pathway development is more influenced by variables unrelated to visual experience (GL). Other variables, including prenatal environment, are also discussed. PMID- 19810803 TI - AGB1 and PMR5 contribute to PEN2-mediated preinvasion resistance to Magnaporthe oryzae in Arabidopsis thaliana. AB - Rice blast, caused by Magnaporthe oryzae, is a devastating disease of rice (Oryza sativa). The mechanisms involved in resistance of rice to blast have been studied extensively and the rice-M. oryzae pathosystem has become a model for plant microbe interaction studies. However, the mechanisms involved in nonhost resistance (NHR) of other plants to rice blast are still poorly understood. Here, we investigated interactions between Arabidopsis thaliana and M. oryzae to identify the genetic basis of NHR. In A. thaliana accessions, preinvasion resistance to M. oryzae in Col-0 was stronger than that of Ler. To examine the genetic basis underlying the natural variation in the responses, we used a well established set of recombinant inbred lines (RIL) derived from a Col x Ler cross and identified three quantitative trait loci that govern the expression of NHR in A. thaliana against M. oryzae. Among the penetration (pen) mutants, only the pen2 mutant allowed increased penetration into epidermal cells by M. oryzae. Double mutant analysis indicated that AGB1 and PMR5 contribute to PEN2-mediated preinvasion resistance to M. oryzae in A. thaliana, suggesting a complex genetic network regulating the resistance. Our results demonstrate that A. thaliana can be used to study mechanisms of NHR to M. oryzae. PMID- 19810804 TI - Pseudomonas syringae pv. tomato DC3000 type III effector HopAA1-1 functions redundantly with chlorosis-promoting factor PSPTO4723 to produce bacterial speck lesions in host tomato. AB - The ability of Pseudomonas syringae pv. tomato DC3000 to cause bacterial speck disease in tomato is dependent on the injection, via the type III secretion system, of approximately 28 Avr/Hop effector proteins. HopAA1-1 is encoded in the conserved effector locus (CEL) of the P. syringae Hrp pathogenicity island. Transiently expressed HopAA1-1 acts inside Saccharomyces cerevisiae and plant cells to elicit cell death. hopAA1 homologs were cloned and sequenced from the CEL of seven P. syringae strains representing diverse pathovars. Analysis of the sequences revealed that HopAA1-1 carries a potential GTPase-activating protein (GAP) domain, GALRA, which is polymorphic (FEN instead of LRA) in HopAA1-2, a paralogous DC3000 effector. Deleting hopAA1-1 from DC3000 reduces the formation of necrotic speck lesions in dip-inoculated tomato leaves if effector-gene cluster IX or just PSPTO4723 within this region has been deleted. A HopAA1-1 mutant in which the putative catalytic arginine in the GAP-like domain has been replaced with alanine retains its ability to kill yeast and promote the formation of speck lesions by the DeltahopAA1-1DeltaIX mutant, but a HopAA1-1 mutant carrying the FEN polymorphism loses both of these abilities. Unexpectedly, PSPTO4723 does not appear to encode an effector and its deletion also reduces disease-associated chlorosis. PMID- 19810805 TI - Agrobacterium-mediated T-DNA transfer and integration by minimal VirD2 consisting of the relaxase domain and a type IV secretion system translocation signal. AB - The VirD2 protein of Agrobacterium tumefaciens is essential for processing and transport of the T-DNA. It has at least three functional domains: a relaxase domain at the N terminus, a bipartite nuclear localization signal (NLS), and a sequence called omega at the C terminus. We confirm here that deletions of the C terminal part of VirD2 led to lack of transfer of T-DNA but, for the first time, we report that virulence is restored when these truncations are supplemented at the C terminus by a short translocation signal from the VirF protein. The lack of virulence of C-terminal deletions suggests that the C-terminal part contains all or part of the translocation signal of VirD2. Using a novel series of mutant VirD2 proteins, the C-terminal half of VirD2 was further investigated. We demonstrate that the C-terminal 40 amino acids of VirD2, which include the NLS and omega, contain all or part of the translocation domain necessary for transport of VirD2 into plant cells, while another element is present in the middle of the protein. The finding that a type IV secretion system transport signal at the C terminus of VirD2 is necessary for virulence provides evidence for the role of VirD2 as a pilot protein driving translocation of the T-strand. PMID- 19810806 TI - Transcriptome analysis of a wheat near-isogenic line pair carrying Fusarium head blight-resistant and -susceptible alleles. AB - Fusarium head blight (FHB), caused primarily by Fusarium graminearum, decreases grain yield and quality in wheat and barley. Disease severity, deoxynivalenol (DON), fungal biomass, and transcript accumulation were examined in a wheat near isogenic line pair carrying either the resistant or susceptible allele for the chromosome 3BS FHB-resistance quantitative trait locus (Fhb1). Fhb1 restricts spread of disease symptoms but does not provide resistance to initial infection or initial DON accumulation. Wheat exhibits both induction and repression of large sets of gene transcripts during F. graminearum infection. In addition, a difference in the general timing of transcript accumulation in plants carrying either the resistant or susceptible allele at the Fhb1 locus was detected, and 14 wheat gene transcripts were detected that exhibited accumulation differences between the resistant and susceptible alleles. These results indicate that these may be host responses that differentiate the resistant from the susceptible interaction. Comparative analysis of the wheat-F. graminearum and the barley-F. graminearum interactions revealed a large set of conserved transcript accumulation patterns. However, we also detected gene transcripts that were repressed in wheat but not in barley. Based on the disease symptoms, transcript accumulation data, and comparative analysis of the barley and wheat host response to F. graminearum infection, we developed an integrated model for the interactions of wheat and barley with F. graminearum. PMID- 19810807 TI - The two conserved cysteine residues of the triple gene block protein 2 are critical for both cell-to-cell and systemic movement of Bamboo mosaic virus. AB - The triple gene block protein 2 (TGBp2) of Bamboo mosaic virus (BaMV) is a transmembrane protein which is known to be required for the cell-to-cell movement of potexviruses. This protein has two conserved Cys residues, Cys-109 and Cys 112, at its C-terminal tail, which is supposed to be exposed on the outer surface of the endoplasmic reticulum (ER) membrane and ER-derived granular vesicles. In this study, we investigated the importance of these two Cys residues on the cell to-cell and systemic movement of BaMV. Our results indicate that the Cys-to-Ala substitutions in TGBp2 make the cell-to-cell movement of BaMV relatively inefficient and the systemic movement of BaMV severely inhibited. Moreover, the defect in systemic movement is attributed to the inefficient transport of viral RNA in the phloem of petiole. Clearly, TGBp2 is critical not only for the cell-to cell but also for the systemic movement of BaMV. In addition, the conserved Cys residues are important for the functioning of TGBp2. PMID- 19810808 TI - The pepper calmodulin gene CaCaM1 is involved in reactive oxygen species and nitric oxide generation required for cell death and the defense response. AB - Calcium signaling has emerged as an important signal transduction pathway of higher plants in response to biotic and abiotic stresses. Ca2+-bound calmodulin (CaM) plays a critical role in decoding and transducing stress signals by activating specific targets. Here, we isolated and functionally characterized the pathogen-responsive CaM gene, Capsicum annuum calmodulin 1 (CaCaM1), from pepper (C. annuum) plants. The cellular function of CaCaM1 was verified by Agrobacterium spp.-mediated transient expression in pepper and transgenic overexpression in Arabidopsis thaliana. Agrobacterium spp.-mediated transient expression of CaCaM1 activated reactive oxygen species (ROS), nitric oxide (NO) generation, and hypersensitive response (HR)-like cell death in pepper leaves, ultimately leading to local acquired resistance to Xanthomonas campestris pv. vesicatoria. CaCaM1 overexpression (OX) Arabidopsis exhibited enhanced resistance to Pseudomonas syringae and Hyaloperonospora parasitica, which was accompanied by enhanced ROS and NO generation and HR-like cell death. Treatment with the calcium-channel blocker suppressed the oxidative and NO bursts and HR-like cell death that were triggered by CaCaM1 expression in pepper and Arabidopsis, suggesting that calcium influx is required for the activation of CaCaM1-mediated defense responses in plants. Upon treatment with the CaM antagonist, virulent P. syringae pv. tomato induced NO generation was also compromised in CaCaM1-OX leaves. Together, these results suggest that the CaCaM1 gene functions in ROS and NO generation are essential for cell death and defense responses in plants. PMID- 19810809 TI - Identification of six type III effector genes with the PIP box in Xanthomonas campestris pv. campestris and five of them contribute individually to full pathogenicity. AB - Xanthomonas campestris pv. campestris is the pathogen of black rot of cruciferous plants. The pathogenicity of the pathogen depends on the type III secretion system (T3SS) that translocates directly effector proteins into plant cells, where they play important roles in the molecular interaction between the pathogen and its hosts. The T3SS of Xanthomonas spp. is encoded by a cluster of hypersensitive response and pathogenicity (hrp) genes. It has been demonstrated that the expression of hrp genes and some type III secreted (T3S)-effector genes is coactivated by the key hrp regulatory protein HrpX. The regulation by HrpX can be mediated by the binding of HrpX protein to a cis-regulatory element named the plant-inducible promoter (PIP) box present in the promoter region of HrpX regulated genes. A genome screen revealed that X. campestris pv. campestris 8004 possesses 56 predicted genes with the PIP box. Nine of these genes have been shown to encode T3S effectors, Hrp, and Hrp-associated proteins. In this study, we employed an established T3S effector translocation assay with the hypersensitive-reaction-inducing domain of X. campestris pv. campestris AvrBs1 as a reporter to characterize the remaining 47 genes with the PIP box and showed that 6 of them, designated as XopXccE1, XopXccP, XopXccQ, XopXccR1, XopXccLR, and AvrXccB, harbor a functional translocation signal in their N-terminal regions, indicating that they are T3S effectors of X. campestris pv. campestris. We provided evidence to demonstrate that all these effectors are expressed in an HrpX-dependent manner and their translocation into plant cells relies on the translocon protein HrpF and the chaperone HpaB. Mutational analyses demonstrated that all these effectors, except AvrXccB, are individually required for full virulence and growth of X. campestris pv. campestris in the host plant Chinese radish. PMID- 19810810 TI - SOD1-targeted gene disruption in the ericoid mycorrhizal fungus Oidiodendron maius reduces conidiation and the capacity for mycorrhization. AB - The genome sequences of mycorrhizal fungi will provide new opportunities for studying the biology and the evolution underlying this symbiotic lifestyle. The generation of null mutants at the wild-type loci is one of the best methods for gene-function assignment in the post-genomic era. To our knowledge, the generation of superoxide dismutase 1 (SOD1)-null mutants in the ericoid mycorrhizal fungus Oidiodendron maius is the first example of a gene-targeted disruption via homologous recombination in a mycorrhizal fungus. The disruption of OmSOD1 by Agrobacterium-mediated transformation resulted in the presence of oxidative stress markers, even in the absence of external superimposed stresses, and an increased sensitivity to reactive oxygen species (ROS)-generating substances, especially to menadione. A reduction in conidiation and in the percentage of mycorrhization of Vaccinium myrtillus roots was also observed. The latter findings establish the pivotal role of SOD1 as an important factor in the relationship between O. maius and its symbiotic partner. The lack of this ROS scavenger may cause an imbalance in the redox homeostasis during host colonization and an alteration in the delicate dialogue between the fungus and its host plant. PMID- 19810811 TI - Genetic analysis of the rkp-3 gene region in Sinorhizobium meliloti 41: rkpY directs capsular polysaccharide synthesis to KR5 antigen production. AB - Rhizobial surface polysaccharides, including capsular polysaccharides (KPS), are involved in symbiotic infection. The rkp-3 locus of Sinorhizobium meliloti 41 is responsible for the production of pseudaminic acid, one of the components of the KR5 antigen, a strain-specific KPS. We have extended the sequence determination and genetic dissection of the rkp-3 region to clarify the structure and function of the rkpY gene and to identify additional rkp genes. Except for rkpY, no other genes were found where mutation affected the KPS structure and symbiosis. These mutants show a unique phenotype producing a low molecular weight polysaccharide (LMW PS). Creating double mutants, we have shown that biosynthesis genes of the KR5 antigen except rkpZ are not necessary for the production of this LMW PS. Polysaccharide analysis of genetically modified strains suggests that rkpY has pleiotropic effects on polysaccharide production. It directs KPS synthesis to the KR5 antigen and influences lipo-oligo 3-deoxy-d-manno-2 octulosonic acid (Kdo) production in S. meliloti 41. In addition, rkpY suppresses the lipo-oligoKdo production when it is introduced into S. meliloti 1021. PMID- 19810812 TI - Transcriptional changes and oxidative stress associated with the synergistic interaction between Potato virus X and Potato virus Y and their relationship with symptom expression. AB - Many virus diseases of economic importance to agriculture result from mixtures of different pathogens invading the host at a given time. This contrasts with the relatively scarce studies available on the molecular events associated with virus host interactions in mixed infections. Compared with single infections, co infection of Nicotiana benthamiana with Potato virus X (PVX) and Potato virus Y (PVY) resulted in increased systemic symptoms (synergism) that led to necrosis of the newly emerging leaves and death of the plant. A comparative transcriptional analysis was undertaken to identify quantitative and qualitative differences in gene expression during this synergistic infection and correlate these changes with the severe symptoms it caused. Global transcription profiles of doubly infected leaves were compared with those from singly infected leaves using gene ontology enrichment analysis and metabolic pathway annotator software. Functional gene categories altered by the double infection comprise suites of genes regulated coordinately, which are associated with chloroplast functions (downregulated), protein synthesis and degradation (upregulated), carbohydrate metabolism (upregulated), and response to biotic stimulus and stress (upregulated). The expressions of reactive oxygen species-generating enzymes as well as several mitogen-activated protein kinases were also significantly induced. Accordingly, synergistic infection induced a severe oxidative stress in N. benthamiana leaves, as judged by increases in lipid peroxidation and by the generation of superoxide radicals in chloroplasts, which correlated with the misregulation of antioxidative genes in microarray data. Interestingly, expression of genes encoding oxylipin biosynthesis was uniquely upregulated by the synergistic infection. Virus-induced gene silencing of alpha-dioxygenase1 delayed cell death during PVX-PVY infection. PMID- 19810813 TI - The absence of Nops secretion in Sinorhizobium fredii HH103 increases GmPR1 expression in Williams soybean. AB - Sinorhizobium fredii HH103 secretes through the type III secretion system at least eight nodulation outer proteins (Nops), including the effector NopP. These proteins are necessary for an effective nodulation of soybean. In this work, we show that expression of the nopP gene depended on flavonoids and on the transcriptional regulators NodD1 and TtsI. Inactivation of nopP led to an increase in the symbiotic capacity of S. fredii HH103 to nodulate Williams soybean. In addition, we studied whether Nops affect the expression of the pathogenesis-related genes GmPR1, GmPR2, and GmPR3 in soybean roots and shoots. In the presence of S. fredii HH103, expression of pathogenesis-related (PR) gene PR1 was induced in soybean roots 4 days after inoculation and it increased 8 days after inoculation. The absence of Nops provoked a higher induction of PR1 in both soybean roots and shoots, suggesting that Nops function early, diminishing plant defense responses during rhizobial infection. However, the inactivation of nopP led to a decrease in PR1 expression. Therefore, the absence of NopP or that of the complete set of Nops seems to have opposite effects on the symbiotic performance and on the elicitation of soybean defense responses. PMID- 19810814 TI - Hexanoic acid-induced resistance against Botrytis cinerea in tomato plants. AB - We have demonstrated that root treatment with hexanoic acid protects tomato plants against Botrytis cinerea. Hexanoic acid-induced resistance (Hx-IR) was blocked in the jasmonic acid (JA)-insensitive mutant jai1 (a coi1 homolog) and in the abscisic acid (ABA)-deficient mutant flacca (flc). Upon infection, the LoxD gene as well as the oxylipin 12-oxo-phytodienoic acid and the bioactive molecule JA-Ile were clearly induced in treated plants. However, the basal ABA levels were not altered. Hexanoic acid primed callose deposition against B. cinerea in a cultivar-dependent manner. Treated plants from Ailsa Craig, Moneymaker, and Rheinlands Ruhm showed increased callose deposition but not from Castlemart. Hexanoic acid did not prime callose accumulation in flc plants upon B. cinerea infection; therefore, ABA could act as a positive regulator of Hx-IR by enhancing callose deposition. Furthermore, although hexanoic acid protected the JA deficient mutant defensless1 (def1), the priming for callose was higher than in the wild type. This suggests a link between JA and callose deposition in tomato. Hence, the obtained results support the idea that callose, oxylipins, and the JA signaling pathway are involved in Hx-IR against B. cinerea. Moreover our data support the relevance of JA-signaling for basal defense against this necrotroph in tomato. Hexanoic acid also protected against Pseudomonas syringae, indicating a broad-spectrum effect for this new inducer. PMID- 19810815 TI - RNA interference highlights the role of CCaMK in dissemination of endosymbionts in the Aeschynomeneae legume Arachis. AB - In legume-rhizobia symbiosis, Ca2+/calmodulin-dependent protein kinase (CCaMK) is essential for rhizobial invasion through infection threads in the epidermis and nodule organogenesis in the cortex. Though CCaMK is actively transcribed in the infected zone of nodules, its role in the later stages of nodule development remain elusive because of the epidermal arrest of "loss-of-function" mutants. In Aeschynomeneae legumes such as Arachis hypogea, rhizobia directly access the cortex, where nodule organogenesis as well as endosymbiont dissemination take place by multiplication of infected cortical cells. We characterized CCaMK (GI:195542474) from A. hypogea and downregulated the kinase through RNA interference (RNAi) to understand its role during organogenesis of its characteristic aeschynomenoid nodules. In CCaMK downregulated plants, the inception of nodules was delayed by approximately 4 weeks and nodulation capacity was decreased (>90%). The infected zones of the RNA interference nodules were scattered with uninfected or binucleated cells as opposed to the homogeneous infection zone in empty-vector-transformed nodules. Symbiosomes in RNAi nodules were pleomorphic with diverse geometrical shapes or arrested during division in the final stages of their fission as opposed to uniform-sized, spherical symbiosomes in empty-vector-transformed nodules. Together, our results reveal CCaMK to be essential for development of functional aeschynomenoid nodules, with a critical role in rhizobial dissemination during nodule organogenesis. PMID- 19810816 TI - IMPDH2 genetic polymorphism: a promoter single-nucleotide polymorphism disrupts a cyclic adenosine monophosphate responsive element. AB - Inosine 5'-monophosphate dehydrogenase (IMPDH), which catalyzes a key step in the de novo biosynthesis of guanine nucleotide, is mediated by two highly conserved isoforms, IMPDH1 and IMPDH2. In this study, IMPDH2 genetic polymorphism was investigated in 96 individuals of Caucasian origin. Four single-nucleotide polymorphisms were identified, comprising one previously described single base pair substitution in the close vicinity of the consensus donor splice site of intron 7 (IVS7+10T>C), and three novel polymorphisms, one silent substitution in exon 9 (c.915C>G), one single base-pair insertion (g.6971_6972insT) within the 3' untranslated region of the gene, and one substitution located in the promoter region (c.-95T>G) in a transcription factor binding site CRE(A) (cyclic adenosine monophosphate [cAMP] response element). Considering the nature and location of this latter polymorphism, its functional relevance was examined by transfecting HEK293 and Jurkat cell lines with constructs of the related region of IMPDH2/luciferase reporter gene. The c.-95T>G mutation leads to a significant decrease of luciferase activity (HEK293: 55% decrease, p < 0.05; Jurkat: 65% decrease, p < 0.05) compared with the wild-type promoter sequence and, therefore, is likely to determine interindividual differences in IMPDH2 transcriptional regulation. These results might contribute to a better understanding of the variability in clinical outcome and dose adjustments of certain immunosuppressors that are metabolized through the IMPDH pathway or that are IMPDH inhibitors. PMID- 19810817 TI - An unusual melting curve on a LightCycler due to a new mutation in methylenetetrahydrofolate reductase. AB - Methylenetetrahydrofolate reductase (MTHFR; E.C. 1.5.1.20) catalyzes the reduction of 5,10-methylenetetrahydofolate to 5-methytetrahydrofolate, and its activity is affected by a number of polymorphisms and mutations, the best known being MTHFR c.677C > T (OMIM 607093.003). The c.677C > T mutation is often analyzed by melting curve analysis after real-time polymerase chain reaction. We observed in one patient upon dual hybridization probe analysis in a Roche LightCycler 1.2 an unusual melting curve peak at 53.4 degrees C instead of one expected at either 51.6 degrees C or 60.4 degrees C. We sequenced the region of MTHFR interrogated by the dual hybridization probes, which revealed a c.685A > G base substitution that would result in an Ile225Val substitution. In conclusion, we identified the cause of an unusual melting curve in a commonly used genotyping technique for MTHFR c.677C > T as due to a novel mutation highlighting the ability of melting analysis to identify new sequence variants. PMID- 19810818 TI - Combined hepatic lipase -514C/T and cholesteryl ester transfer protein I405V polymorphisms are associated with the risk of coronary artery disease. AB - Hepatic lipase (LIPC) and cholesteryl ester transfer protein (CETP) are important components of high-density lipoprotein (HDL) metabolism and reverse cholesterol transport. Therefore, their genes are promising candidate genes for cardiovascular disease. The aim of the present study was to investigate whether combined LIPC -514C/T and CETP I405V polymorphisms correlate with angiographically documented coronary artery disease (CAD). Genotyping was performed in 317 patients who underwent clinically indicated coronary angiography. The patients were classified with significantly diseased arteries if one or more coronary arteries had a stenosis >50% and with minimally diseased arteries if there was no significant stenosis (<40%) in any artery. There were no significant associations of individual polymorphisms with the risk of significant CAD. In a multivariate logistic regression analysis including cardiovascular risk factors, simultaneous presence of both LIPC -514T and CETP 405V alleles was an independent predictor of significantly diseased arteries (odds ratio = 2.04; p = 0.022). This association was not significant in women with combined genotype who had the highest HDL-cholesterol. In conclusion, the combined T allele of LIPC 514C/T and V allele of CETP I405V are associated with the risk of CAD. Further, the higher HDL-cholesterol and female gender may reduce the effect of combined genotype on CAD risk. PMID- 19810819 TI - Significant genetic differentiation within the population of the Island of Corsica (France) revealed by y-chromosome analysis. AB - Using 10 Y-chromosome short tandem repeat allelic and haplotypic frequencies, we examined genetic variation within the population of Corsica and its relationship with other Mediterranean populations. The most significant finding is the high level of genetic differentiation within Corsica, with strong evidence of an effective barrier to male-mediated gene flow between the south and the rest of the island. This internal differentiation most probably results from low exogamy among small isolated populations and also from the orography of the island, with a central mountain chain running the length of the island restricting human movement. This physical barrier is reflected not only in present-day intraisland linguistic and genetic differences but also in the relatedness of Corsican regions to other Mediterranean groups. Northwest and Central Corsica are much closer to West Mediterranean populations, whereas South Corsica is closer to Central-North Sardinia and East Mediterranean populations. PMID- 19810820 TI - Significance of genome-wide association studies in molecular anthropology. AB - The successful advent of a genome-wide approach in association studies raises the hopes of human geneticists for solving a genetic maze of complex traits especially the disorders. This approach, which is replete with the application of cutting-edge technology and supported by big science projects (like Human Genome Project; and even more importantly the International HapMap Project) and various important databases (SNP database, CNV database, etc.), has had unprecedented success in rapidly uncovering many of the genetic determinants of complex disorders. The magnitude of this approach in the genetics of classical anthropological variables like height, skin color, eye color, and other genome diversity projects has certainly expanded the horizons of molecular anthropology. Therefore, in this article we have proposed a genome-wide association approach in molecular anthropological studies by providing lessons from the exemplary study of the Wellcome Trust Case Control Consortium. We have also highlighted the importance and uniqueness of Indian population groups in facilitating the design and finding optimum solutions for other genome-wide association-related challenges. PMID- 19810821 TI - The p.Arg258Gly mutation in intracellular loop 2 of CFTR is associated with CFTR related disorders. AB - Missense mutations account for approximately 50% of the mutations described in the CFTR gene. However, their proportion is higher in CFTR-related disorders (CFTR-RD) than in cystic fibrosis (CF), suggesting a different mutational spectrum. The uncertainty surrounding many of these mutations prevents suitable genetic counseling. Thus, it is crucial to determine whether a missense mutation has clinical expression, and if it does, to then define the associated phenotype. Herein we have assessed the phenotype associated with the p.Arg258Gly (R258G) mutation, checking our cohorts of patients (CF and CFTR-RD) and control subjects (CF carriers, fertile males, and general population). We also performed in silico predictive studies on the possible consequences of this mutation at the protein level. Lastly, we exhaustively reviewed the literature on this mutation. To date, R258G has only been found in six patients: a French congenital bilateral absence of vas deferens patient, reported in 1995 and five unrelated subjects from our cohort of non-CF patients, described here. Based on these findings, we postulate that R258G is primarily a CFTR-RD-associated mutation. PMID- 19810822 TI - Single-nucleotide polymorphisms in genes encoding toll-like receptor -2, -3, -4, and -9 in case-control study with breast cancer. AB - BACKGROUND: Genetic susceptibility to cancer is multifactorial, and it is known that impairment of the immune system could contribute to risk for getting cancer. AIM OF THE STUDY: Single-nucleotide polymorphisms (SNPs) of Toll-like receptor (TLR) 2, TLR3, TLR4, and TLR9 genes, which are important for innate immunity, were analyzed for the association with breast cancer. METHODS: The SNPs comprised TLR2 (c.597T>C), TLR2 (c.1350T>C), TLR3 (c.1377C>T), TLR4 (c.896A>G), and TLR9 (c.1635A>G). The allelic and genotypic frequencies of these TLR SNPs were compared between patients (n = 130) and controls (n = 101) in a case-control study from Croatia. RESULTS: TLR SNPs were not significantly different. From the population genetics viewpoint, we found that a hypomorphic variant of TLR4 (p.Asp299Gly) allele has no specific allelic frequency (8.4%) in the Croatian population (n = 496) compared to other Caucasians (6.5-10%). CONCLUSION: These results suggest that polymorphisms in tested TLR genes are not likely to be associated with increased risk for developing breast cancer. PMID- 19810823 TI - Presymptomatic diagnosis in Huntington's disease: the Mexican experience. AB - Huntington's disease (HD) is an autosomal dominant progressive, disabling neurodegenerative disorder, for which there is no effective treatment. Predictive testing (PT) for this illness began in 1986 and by 1993 it became more precise after cloning of the gene and the discovery of a CAG repeat expansion as the underlying cause. The objective of this paper is to illustrate the implementation and results of a PT program in a group of at-risk Mexican individuals with 12 years of follow-up. Our PT program conforms to the guidelines proposed by the International Huntington Association and the HD Working group of the World Federation of Neurology. Seventy-five individuals requested the testing, four of them did not fulfill the inclusion criteria, and five abandoned the program voluntarily before receiving the test results. Therefore, 66 results were delivered to 41 noncarriers and 25 mutation carriers. We did not have any catastrophic event, but 4 individuals with normal results and 11 mutation carriers were depressed. Even if this is a small sample, it is the first report of PT in a Latin-American population in which we have been faced with the same problems referred to in larger series. PMID- 19810824 TI - Methylenetetrahydrofolate reductase and angiotensin-converting enzyme gene polymorphisms among Saudi population from Qassim region. AB - BACKGROUND: Hypertension and cardiac diseases are multifactorial disorders with genetic background determined by multiple gene polymorphisms. METHODS: This work included 273 healthy unrelated subjects ethnically belonging to the Qassim region. Their age ranged from 18 to 60 years, with a median age of 20 years. They included 152 (55.7%) men and 121 (44.3%) women. Their DNAs were analyzed for genetic polymorphisms of the methylenetetrahydrofolate reductase (MTHFR; 677C/T and 1298 A/C) as well as angiotensin-converting enzyme genes (ACE; insertion/deletion [I/D]) using real-time polymerase chain reaction. RESULTS: Carriers of the mutant MTHFR 677 T allele (CT + TT) and that of the 1298 C allele (CC + AC) constituted 33.7% and 48.9% of studied subjects, respectively, whereas carriers of ACE gene mutant D allele (DD + ID) represented 93.3% of subjects. The allele frequencies of MTHFR 677T, 1298C, and ACE D alleles were 18.7%, 29.45%, and 72.5%, respectively. Haplotype analysis of characterized chromosomes revealed that 2.5% were likely to carry the three mutant alleles together, 30.91% were likely to carry two of the three mutant alleles, and 51.92% were likely to carry one mutant allele. CONCLUSION: The Saudi population from the Qassim region is a carrier of a relatively large number of genetic alleles predisposing them to hypertension and cardiac diseases. This gives a warning to local health authorities for adoption of competent programs for prevention as well as early diagnosis and management. PMID- 19810825 TI - Attitudes toward genetic testing in a german population. AB - AIMS: Population-representative or population-based studies on attitudes toward molecular genetic testing have so far been conducted in the United States, Great Britain, the Netherlands, and Finland. In 2001 our group conducted a representative survey for a German population among a total of 2076 respondents aged 14-95 years. The questionnaire administered consisted of 13 items covering the advantages and disadvantages of molecular genetic testing for hereditary diseases. We conducted a factor analysis to explore potential sub-dimensions of attitudes, and describe here the influence of sociodemographic variables on these sub-dimensions and how the attitudes of the Germans compare to those of the Finns. RESULTS: Our factor analysis of the results showed three dimensions: approval, disapproval, and concern regarding genetic testing. In the German sample we found an overall positive attitude toward genetic testing. There was no influence of sex, but there were marked differences regarding age, educational backgrounds, and religious affiliation. Finns are more in favor of genetic testing and disapprove less than Germans. We can summarize that Germans have a markedly more skeptical view than Finns. In our opinion this could well be understood in the context of German history. We assume that this created a less conducive environment for the population to form a clearer attitude toward genetic testing as compared to the Finns. CONCLUSIONS: In the light of our results there is certainly a need for more information and education. Further, it would in our opinion be most useful if the German development regarding attitudes to genetic testing, especially after reunification, is studied longitudinally to provide a better understanding of possible developments. PMID- 19810826 TI - High-risk fragile x screening in Guatemala: use of a new blood spot polymerase chain reaction technique. AB - BACKGROUND: Because fragile X syndrome (FXS) is prevalent, it has become the subject of newborn and high-risk screening efforts. International screening, however, can be financially and logistically prohibitive, particularly in countries where resources may be scarce. Recently, we have developed a screening test on blood spot that can detect expanded alleles from the normal through the full mutation range in both males and females. It is accurate, rapid, inexpensive, and applicable on blood spots and therefore ideal for international screening. The use of this blood spot screening technique was piloted in "a high risk screening" study of individuals in Guatemala. METHODS: One hundred and five blood spots from subjects from Guatemala were screened for the Fragile X Mental Retardation 1 mutation. They were classified as "high-risk" through placement into one of the following five categories: (a) relatives of someone with a previous FXS diagnosis, (b) individuals with confirmed autism, (c) individuals with confirmed intellectual disability, (d) individuals with Parkinson's-like presentation, and (e) individuals with a family history of intellectual disability but no confirmed cases of FXS. RESULTS: Fifteen of the individuals tested yielded an expanded allele, 10 premutations and 5 full mutations. All 15 expansions were found in individuals with a relative with a confirmed FXS diagnosis. No expansions were found in the other clinical groups. CONCLUSIONS: Blood spot polymerase chain reaction screening is an effective, cost-efficient method to conduct cascade testing in families with a known history of FXS, even in small screening cohorts. PMID- 19810830 TI - Does soy-based infant formula cause ADHD? PMID- 19810827 TI - Emerging infections in burns. AB - BACKGROUND: Patients who suffer severe burns are at higher risk for local and systemic infections. In recent years, emerging resistant pathogens have forced burn care providers world wide to search for alternative forms of treatment. Multidrug-resistant Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter spp., and various fungal strains have been the major contributors to the increase in morbidity and mortality rates. Multi-drug-resistant S. aureus remains the major cause of gram-positive burn wound infections world wide. Treatment strategies include rigorous isolation protocols and new types of antibiotics where necessary. METHODS: We reviewed 398 severely burned patients (burns >40% total body surface area [TBSA]) admitted to our hospital between 2000 and 2006. Patients who did not contract multi-drug-resistant gram-negative organisms during their hospital course and received our standard antibiotic regimen-vancomycin and piperacillin/tazobactam-served as controls (piperacillin/tazobactam; n = 280). The treatment group consisted of patients who, during their acute hospital stay, developed infections with multi-drug-resistant gram-negative pathogens and were treated with vancomycin and colistin for at least three days (colistin; n = 118). RESULTS: Gram-negative organisms continue to cause the most severe infections in burn patients. Colistin has re-emerged as a highly effective antibiotic against multiresistant Pseudomonas and Acinetobacter infections of burns. Patients who required colistin therapy had a significantly larger average total and full thickness burn than patients treated with piperacillin/tazobactam and vancomycin, and the mortality rate was significantly higher in the colistin group (p < 0.05). However, there was no significant difference between the colistin and piperacillin/tazobactam groups in the incidence of neurotoxicity, hepatic toxicity, or nephrotoxicity. The main fungal pathogens in burn patients are Candida spp., Aspergillus spp., and Fusarium spp. A definitive diagnosis is more difficult to obtain than in bacterial infections. Amphotericin B and voriconazole remain the two most important anti-fungal substances in our practice. CONCLUSIONS: Innovations in fluid management, ventilatory support, surgical care, and antimicrobial therapy have contributed to a significant reduction in morbidity and mortality rates in burn patients. Vancomycin and clindamycin are the two most important reserve antibiotics for methicillin-resistant Staphylococcus aureus infection. Oxazolidinones and streptogramins have showed high effectiveness against gram-positive infections. Colistin has re-emerged as a highly effective antibiotic against multiresistant Pseudomonas and Acinetobacter infections. Current challenges include Candida, Aspergillus, and molds. The development of new agents, prudent and appropriate use of antibiotics, and better infection control protocols are paramount in the ongoing battle against multi resistant organisms. PMID- 19810832 TI - Clinical study results of tolterodine in patients with overactive bladder. AB - Overactive bladder is a chronic condition that affects millions of people worldwide. Antimuscarinic agents are the predominant pharmacotherapy for overactive bladder. Tolterodine is a potent, competitive muscarinic receptor antagonist that exhibits in vivo selectivity for the bladder over other tissues that contain muscarinic receptors (such as the salivary glands and the eye), which helps explain why it has such a good tolerability profile compared to oxybutynin. Tolterodine is the first drug specifically developed for the treatment of overactive bladder and is available as both an immediate- and extended-release formulation. This review summarizes the primary antimuscarinic profile of tolterodine and reviews the efficacy, safety and tolerability of tolterodine in studies of patients with overactive bladder. PMID- 19810833 TI - Amitriptyline in the treatment of primary headaches. AB - Prophylactic treatment of primary headaches should be considered if the patient has severe or frequent attacks and if management of the acute attack is inadequate. The tricyclic antidepressant, amitriptyline, is widely used for the prophylactic treatment of migraine and tension-type headache. In spite of its extensive use, amitriptyline has only been studied in few controlled trials in migraine. These studies indicate, but do not prove, that amitriptyline is effective in migraine. Amitriptyline has been shown to be moderately effective in chronic tension-type headache in several controlled studies, but the use is often hampered by side effects. Conclusive studies regarding the efficacy of amitriptyline in migraine are much wanted. There is great need for development of more efficacious drugs with fewer side effects for the prophylactic treatment of primary headaches. PMID- 19810834 TI - Role of acetaminophen in the treatment of acute stroke. AB - The effect of core body and brain temperature on stroke outcomes has been addressed in many animal and human studies. From these, it is clear that elevated temperature worsens ischemic neuronal injury and that hypothermia may be neuroprotective. Based on these data, a few investigators have examined the role of fever prevention and induced hypothermia in the treatment of acute stroke patients. Since mechanical cooling techniques face inherent practical and safety problems, recent attention has been focused on pharmacological means. Specifically, a handful of studies have looked at whether acetaminophen is useful in preventing fever, or inducing hypothermia in stroke patients. These studies demonstrated only marginal effects on body temperature and none were sufficiently powered to address functional outcomes. In this review article, the rationale for acetaminophen therapy in acute stroke patients and the data regarding its potential though modest clinical role, will be discussed. PMID- 19810835 TI - Second-generation antiepileptic medications in children: efficacy and cognitive effects. AB - This paper is from the perspective of a user (prescriber) of antiepileptic medications in children who is attempting to practice evidence-based medicine. This is a review of published data comparing antiepileptic efficacy and cognitive effects from only randomized comparative trials. Data are almost nonexistent in young children, although adolescents are frequently included in the adult trials presented. These studies are inevitably flawed and the ideal study is suggested, but will in reality, never be performed. The review provides enough detail of previous studies so that the reader considering future antieplileptic drug trials can recognize how to avoid many of the problems in an era requiring proof of claims. Comparative studies will allow the supplier of the product (the physician) to make rational choices that can only improve market share. PMID- 19810836 TI - Current therapeutic strategies for anxious depressives. AB - Anxious depressives refers to a group of patients with a primary depressive disorder, who also have significant symptoms of anxiety. Despite the fact that this group of patients is widely recognized and understood to have a poorer prognosis, this group has been poorly studied to date. In this review, we briefly examine the pathophysiology and neurophysiology of this condition. We then reviewed in detail the relevant clinical studies which have been carried out in this condition and published over the last 5 years. Taken together, the results of published studies begin to suggest that patients with anxious depression may respond better to antidepressants that are dual action drugs (such as serotonin and noradrenaline re-uptake inhibitors), rather than single action drugs (such as specific serotonin re-uptake inhibitors). However, further controlled studies are warranted to test this hypothesis. PMID- 19810837 TI - Mood disorders in patients with COPD: overview of current research and future needs. AB - Depression is major problem for the medically ill population but few recommendations have been made regarding treatment with specific compounds in many of these illnesses. This review attempts to consider the inter-relationship between smoking, pulmonary disease and mood disorders. It has been reported that nearly 50% of patients with chronic pulmonary illnesses report depressive symptoms at some time during the course of their illness, but with the complex issues regarding respiratory drive and substance dependence, little in the way of specific clinical information have been made for this group. In this review, the theorectical relationship that exists between dopamine, serotonin and nicotine and how the biochemical nicotine dependence issues (with regard to mood) may actually be a key element in the understanding and treatment of depression later in the life of the development of chronic lung disease and depression, are discussed. PMID- 19810838 TI - Microalbuminuria in cerebrovascular diseases. AB - Microalbuminuria was originally introduced to clinical practice as a useful marker of incipient diabetic nephropathy. The growing body of evidence suggests that increased urinary albumin excretion is related to the increased risk of cardiovascular events and all causes mortality in subjects with or without diabetes. More over, transient increases of albuminuria were found in numerous acute diseases, including trauma, meningitis, myocardial infarction and stroke. Microalbuminuria was found as an important prognostic variable in these settings, but the exact mechanism of its transient occurrence remains unknown. The measurement of daily albumin excretion is a relatively easy and inexpensive diagnostic tool. The predictive value of increased albuminuria is promising but requires further detailed studies. This paper reviews the relevance of microalbuminuria for cerebrovascular diseases, stressing the future research plans and clinical implications. PMID- 19810839 TI - Interventional techniques for the treatment of head and neck pain. AB - Daily head and neck pain is the bain of existence for thousands of people every day. The economic burden is immense. The treatment of migraine headaches alone amounts to billions of dollars per year in direct and indirect costs. Conservative measures help many, but for the unfortunate few their search continues. Thankfully, therapeutic options exist in the hands of invasive pain management physicians. Minimally to highly invasive pain procedures can provide relief that allows the patient some aspect of normality. This article will review available options and support them with recent studies. PMID- 19810840 TI - Treatment of chronic inflammatory demyelinating polyneuropathy. AB - Chronic inflammatory demyelinating polyradiculoneuropathy is a neuromuscular disorder for which immunological factors are undoubtedly important in the etiopathogenesis. As clinical recognition of this disorder has improved, immunosuppressive and immunomodulating treatments have emerged as the treatments of choice and the prognosis for patients with chronic inflammatory demyelinating polyneuropathy has improved. This review discusses the treatments most frequently employed and summarizes the evidence that establishes the beneficial impact these treatments have had on the clinical course of patients with chronic inflammatory demyelinating polyneuropathy. PMID- 19810841 TI - Cerebrospinal fluid and serum antimicroglial antibodies: prospects for early diagnosis of Alzheimer's disease. AB - The acceptance that chronic inflammation plays a role in the pathogenesis of Alzheimer's disease has widen the window of opportunity for novel therapeutics. The need to establish markers to detect early and preclinical symptoms of Alzheimer's disease is critical as this could allow early intervention. Activated microglia, now considered as the immune cell of the CNS, have gained recognition as participating in the cascade of early events leading to Alzheimer's disease pathology. The serendipitous findings of microglia antibodies in the serum and cerebrospinal fluid of Alzheimer's disease patients could be a means to distinguish Alzheimer's disease patients from other dementias. Further investigations revealing the presence of these particular antibodies in patients at earlier stages suggested that they may be a source to monitor the progression of the disorder. Combining the detection of these antibodies with clinical trials could provide essential feedback about the efficacy of the therapies to interfere with the disease process. PMID- 19810843 TI - Bupropion for attention deficit hyperactivity and comorbid disorders. AB - Bupropion is a unique antidepressant with noradrenergic and to a lesser extent dopaminergic effects. These have led investigators to explore bupropion's efficacy in attention deficit hyperactivity disorders, which are believed to be related to aberrations in catecholamines. An expanding body of research has demonstrated bupropion's efficacy in attention deficit hyperactivity disorders over the lifespan. Despite early reports of seizures in select samples, bupropion is generally well-tolerated. While stimulants remain the first-line pharmacological treatments for attention deficit hyperactivity disorders, bupropion offers promise as a medication that treats not only attention deficit hyperactivity disorders, but also common comorbid disorders, including unipolar and bipolar depression, anxiety and substance abuse. PMID- 19810844 TI - Buspirone in Alzheimer's disease. AB - Anxiety symptoms are experienced by the majority of patients with Alzheimer's disease. Generalized anxiety disorder may occur in 5-6% of patients with Alzheimer's disease. Anxiety symptoms may underlie agitation and aggression. Anxiety and agitation cause significant morbidity, caregiver distress and may even precipitate institutionalization. Benzodiazepines, although frequently used to treat anxiety and agitation in patients with Alzheimer's disease, should be avoided because of the high morbidity associated with their use. Disturbances in serotonergic neurotransmission may underlie anxiety symptoms and agitation. Preliminary evidence suggests that buspirone may be a good non sedating alternative to treat Alzheimer's disease patients with persistent anxiety symptoms and agitation-aggression. Effective doses reported range from 15 to 60 mg/day is generally well-tolerated. Large, randomized controlled trials are needed to confirm the potential benefits of buspirone for anxiety symptoms and other behavioral disturbances in Alzheimer's disease patients. PMID- 19810845 TI - Neuronal glucose metabolism and schizophrenia: therapeutic prospects? AB - This review will focus on the treatment and prevention of schizophrenia in children and adolescents. Neurodevelopmental theories suggest that loss of gray matter and defective synaptic function are major etiological factors in this disease. The efficacy of current antipsychotic medications has been discussed, however, these drugs produce serious side effects and may adversely affect the developing brain. We propose a novel therapeutic approach, termed neuroenhancement, that aims to promote neuronal survival and optimize neuronal function through the use of drugs. The goal is to enhance glucose metabolism in the brain, which would support higher functional activity in neurons and provide neuroprotection. Future drug development for the treatment of childhood schizophrenia should focus more on optimization of neuronal function rather than tranquilization and symptomatic relief. PMID- 19810846 TI - Genetic study of schizophrenia: new targets for therapy. AB - Recent advances in genomic and genetic knowledge and technology have opened the way to deciphering the extremely complex molecular mechanisms underlying schizophrenia. Despite accumulated pharmacological and clinical evidence for the dopamine hypothesis of schizophrenia, genetic studies have shown that monoamine related genes, including dopamine D3 and 5-HT(2A) receptors, are associated with only a modest risk of susceptibility to schizophrenia. However, recent studies have revealed many attractive candidate genes, such as: cytosolic phospholipase A2, potassium intermediate/small conductance calcium-activated channel, subfamily N, member 3, sigma receptor type 1, cannabinoid receptor 1, dihydropyridimdinase related protein 2, proline dehydrogenase 2, Disc-1, dysbindin and neuregulin 1. These molecules are involved in a wide range of brain functions and may become targets for novel therapy of schizophrenia in the immediate future. PMID- 19810847 TI - Population-based gene discovery in psychiatric diseases. AB - There is strong evidence that psychiatric disorders, such as schizophrenia and bipolar, have a significant genetic component. Yet, despite many attempts, the susceptibility genes that predispose to these illnesses have not been revealed. We describe an approach to the discovery of genes involved in such complex diseases. The prospect of gene discovery is optimized by considering elements, such as the applicability of a family- versus population-based paradigm, sample size and use of an isolated population. We suggest that a high level of statistical significance can be achieved by efficient scanning of a dense map of polymorphisms followed by intensive genotyping of informative variations and haplotype analysis. PMID- 19810848 TI - Weight gain associated with atypical antipsychotic drugs: mechanisms and management. AB - Being overweight or obese are medical conditions that are very difficult to treat. There is compelling evidence that obesity is commonly seen in patients with schizophrenia. Recently, a number of publications have focused on the ability of atypical antipsychotic drugs to induce obesity. All antipsychotic drugs produce weight gain, but their potential varies. Many studies overwhelmingly confirm that atypical antipsychotic drugs produce substantially more weight gain in comparison with typical antipsychotic drugs. Clozapine and olanzapine have the most weight-inducing potential. Even ziprasidone, which is considered to be weight neutral, produces weight gain in some patients. The pathophysiology of weight gain is complicated. Many neurohormones, neuropeptides and intestinal hormones, as well as adipose tissue and hair root-derived hormones, interact with environmental factors to produce weight gain. The basis of weight-gain treatment is an understanding of the etiology. Drug-induced obesity provides a unique opportunity to psychiatrists to understand this true psychosomatic problem. In the absence of this knowledge, prevention is the best hope. Education, diet control and simple behavioral measures may prevent excessive weight gain. In those with weight gain, treatment can be attempted with pharmacotherapy. PMID- 19810849 TI - Management of psychiatric side effects associated with corticosteroids. AB - Corticosteroids are given for a wide variety of inflammatory or immune response related illnesses. Dose-dependent psychiatric symptoms, including mania, depression, cognitive impairment and occasionally psychosis, have been reported during corticosteroid therapy. As many patients require long-term therapy with corticosteroids, treatments for these symptoms are sometimes needed. One small controlled study suggested that lithium prevents psychiatric symptoms with corticosteroids. Case reports suggest the efficacy of several different anticonvulsants and antipsychotics. The anticonvulsant lamotrigine may be useful for depression and cognitive impairment associated with corticosteroid therapy. Literature on antidepressant use is mixed, with reports of symptom worsening with tricyclic antidepressants but improvement with selective serotonin reuptake inhibitors. Larger controlled studies are needed to determine optimal treatment for the psychiatric side effects of corticosteroids. PMID- 19810850 TI - Use of antipsychotic medication in chemotherapy-induced nausea and vomiting. AB - Nausea and vomiting continue to present significant problems for cancer patients undergoing chemotherapy. Initial work suggested an important role for central dopamine transmission in the underlying pathophysiology. However, recent evidence has implicated central and peripheral serotonin release. Although the implementation of 5-HT(3) receptor antagonists has resulted in significant improvement in acute symptoms of nausea and vomiting, they have not demonstrated optimal efficacy in anticipatory and delayed emetic syndromes and are significantly more expensive than other antiemetic therapies. The use of typical antipsychotic medication to reduce nausea and vomiting in cancer patients is discussed, as is the potential efficacy of newer atypical antipsychotics, which have activity at receptors implicated in the control of chemotherapy-induced nausea and vomiting and demonstrate an improved side-effect profile. PMID- 19810851 TI - Antipsychotics for aggressive adolescents: barriers to best practice. AB - In adolescents, antipsychotics are most often used to treat complex, comorbid conditions with core disruptive features. However, the literature guiding such practices is limited. Best practice guidelines bridging the gap between the evidence and clinical practice have been developed to promote the appropriate and safe use of antipsychotics in aggressive youths. Due to complex barriers that exist at the level of the physician, patient/family and organization, merely disseminating these guidelines will not likely change antipsychotic prescribing practices. Negative attitudes, time constraints, lack of staff training and resources, or adolescent/family nonadherence can impede the translation of best practice guidelines into routine practices. Efforts to implement best practice guidelines must address these barriers if changes in prescribing practices are to occur and be sustained. PMID- 19810852 TI - Provisional diagnostic criteria for depression of Alzheimer's disease: description and review. AB - This review centers on the development of diagnostic criteria for depression of Alzheimer's disease. It describes: risk-factors and neurobiological correlates, epidemiology, clinical characteristics and course, assessment, treatment, economics, a description of the criteria and future research directions. Overall, there is substantial evidence for depression of Alzheimer's disease. Further research is needed to better define core symptoms, clinical course and efficacy of treatments. PMID- 19810853 TI - The evolution of antiepileptic drugs for mood stabilization and their main mechanisms of action. AB - Although much progress has been made in successfully treating bipolar disorder, there is increasing awareness of the limitations of traditional treatment regimes, such as lithium or antipsychotics and the possible beneficial use of antiepileptic drugs. After the first generation of antiepileptic drugs such as phenytoin and clonazepam, the second generation is comprised of the frequently used substances carbamazepine/oxcarbazepine and valproate. Lamotrigine, gabapentin, tiagabine, levetiracetam, zonisamide and topiramate will represent the third generation 5 years from now. Drugs such as retigabine might represent the next generation. However, the efficacy of antiepileptic drugs investigated in the treatment of bipolar disorder differs and most promising effects are seen in combination therapy with mood stabilizers. The authors review the main mechanisms of action of these drugs which may, in turn, improve our understanding of the pathophysiology of bipolar disorder. PMID- 19810854 TI - Current therapeutic approaches for dysthymic disorder. AB - The purpose of this article is to review the literature regarding current treatments for dysthymic disorder. We will first discuss definitions of dysthymic disorder, demographics of these patients, clinical course and family history. We will then review treatment studies for psychotherapy, pharmacotherapy and combined treatment, discuss treatment guidelines for dysthymic disorder, render an 'expert opinion' regarding problems in the definition of dysthymic disorder, elaborate on areas for future research and speculate on a 'five-year view.' PMID- 19810855 TI - What makes epilepsy drug refractory? AB - About 20-40% of patients with epilepsy will be refractory to medical treatment with antiepileptic drugs. It is unclear whether patients are already drug resistant at the time of their initial presentation, or whether they become so over the course of their illness. Identifying predictors for drug-refractory epilepsy may be important for directing epilepsy patients to an effective nonpharmacological treatment, such as surgery or the vagus nerve stimulator, in a timely manner. In addition, understanding the factors that lead to the drug refractory state may facilitate the development of new therapies that are effective in the resistant subgroup. This paper identifies various predictors that have been associated with drug-refractory epilepsy, discusses the evidence behind each factor and recommends strategies for clarifying predictors of refractoriness. PMID- 19810856 TI - Diagnosis, pathogenesis and treatment of Miller Fisher syndrome and related disorders: clinical significance of antiGQ1b IgG antibody. AB - Miller Fisher syndrome is a variant of Guillain-Barre syndrome, characterized by ophthalmoplegia, ataxia and areflexia. The antiGQ1b immunoglobulin G antibody is a specific marker of Miller Fisher syndrome and related disorders, such as Guillain-Barre syndrome with ophthalmoplegia, atypical Miller Fisher syndrome characterized by acute ophthalmoplegia or acute ataxia and Bickerstaff's brainstem encephalitis. The antiGQ1b immunoglobulin G antibody may play some important roles in the pathogenesis of Miller Fisher syndrome and related disorders. Possible mechanisms are discussed. Molecular mimicry between an infectious agent of the antecedent infection and the ganglioside may be a mechanism of the antibody production. Plasmapheresis or intravenous immunoglobulin therapy may be warranted for Miller Fisher syndrome and Bickerstaff's brainstem encephalitis, as well as Guillain-Barre syndrome with ophthalmoplegia. PMID- 19810857 TI - Current and future therapeutic strategies for Alzheimer's disease and related dementias. PMID- 19810859 TI - Glatiramer acetate for multiple sclerosis: a comprehensive review of mechanisms and clinical efficacy. AB - The 'Decade of the Brain' (1990-2000) saw unprecedented advances in neurosciences including multiple sclerosis. It could have not been more aptly named, as it produced a shift in the paradigm of multiple sclerosis management, making multiple sclerosis a treatable disorder with the availability of several therapeutic options. For a chronic progressive neurological disorder like multiple sclerosis, this change in the understanding and treatment touched the lives of hundreds of thousands of patients worldwide and many more who provided care and counsel as family and friends. Of the four agents available for the treatment of the most common type of multiple sclerosis - relapsing-remitting - three are beta-interferons and one is a noninterferon polypeptide of four amino acids (glatiramer acetate) with a distinct immunomodulating profile. Glatiramer acetate is now approved and available in North America, Europe and many other countries. It has been tested in pivotal trials as well as long term extension trials for almost 10 years (8 years published) providing remarkable evidence of efficacy and safety. This review will highlight the immune mechanisms and clinical data reported with glatiramer acetate in multiple sclerosis over the past three decades. PMID- 19810860 TI - Update on therapies for cluster headache. AB - Cluster headache is one of the primary headache disorders. It is considered the most severe headache syndrome known to humans. In most instances, this disorder is readily treatable when the correct medications are utilized at the correct dosages. Cluster headache treatment involves abortive, transitional and preventive therapy strategies. PMID- 19810861 TI - Key factors in the primary care diagnosis of migraine. AB - Classification systems for migraine and related daily headache syndromes represent expert consensus and lack biologic markers. These diagnostic categories have been essential to standardize research but might not be ideal in primary care settings. New paradigms which may be more useful for primary care physicians are explored. PMID- 19810862 TI - Glucose regulation in headache: implications for dietary management. AB - Dietary factors are frequently cited as 'triggers' of migraine and other types of headache. A review of the literature indicates that two specific dietary factors frequently induce headache: fasting and the relatively mild reactive hypoglycemia that can follow large carbohydrate ingestions. The maintenance of serum glucose levels to supply the energy for the nervous system is a primary responsibility of the sympathetic nervous system. It is suggested that migraine and other headache attacks may, at least in part, result from or be exacerbated by the sympathetic nervous system activation required to provide a steady supply of serum glucose for brain energy needs. Dietary patterns that provide a steady source of serum glucose, while minimizing serum glucose fluctuations, are hypothesized to help prevent and/or treat migraine and other headache attacks. PMID- 19810863 TI - Update on the diagnosis of multiple sclerosis. AB - The diagnosis of multiple sclerosis has been increasingly standardized over the years and has evolved to incorporate new diagnostic modalities. The gold standard for diagnosing multiple sclerosis remains clinical, with dissemination of typical white matter symptoms and signs in time and space. The Schumacher criteria in 1965 attempted to standardize clinical criteria for diagnosing multiple sclerosis. The Poser criteria in 1983 added evoked potential and cerebrospinal parameters and the McDonald criteria in 2001 added MRI parameters. All criteria for diagnosing multiple sclerosis include the caveat that no alternative diagnosis better explains the clinical picture, making the differential diagnosis of multiple sclerosis critical. Recent availability of first generation immunotherapies for MS has increased pressure to make an early and accurate diagnosis of multiple sclerosis and to use the diagnostic work-up to try to prognosticate a future disease course. PMID- 19810864 TI - Genetic susceptibility to multiple sclerosis: implications of genetic research on MS therapy. AB - Multiple sclerosis is a complex disease with a multifactorial etiology. Although environmental risk-factors are clearly involved in the occurrence of multiple sclerosis, the importance of genetic factors has been strongly supported by the results of previous reports. Here, we review the results of recent studies and hypotheses concerning genes related to multiple sclerosis and the modifying role of different genes in the course and the severity of multiple sclerosis. To date, these data have not been considered in the therapy of multiple sclerosis. We describe the applicability of these results toward designing specific therapies for multiple sclerosis. PMID- 19810865 TI - Childhood multiple sclerosis: course and management. AB - Multiple sclerosis in children and adolescents carries clinical and prognostic differences from the disease in adults. Although new disease-modifying therapeutic agents which became available in recent years have improved the course and prognosis of multiple sclerosis, the indications and use of these drugs in childhood have not been evaluated systematically. In addition, studies of the complex immunopathology of the disease provide many new therapeutic possibilities to be investigated. While early treatment is preferred because immunomodulatory drugs appear more efficient in early disease, the insufficiency of the current knowledge about the safety, dosage and adverse effects of new drugs in children restrict clinicians in their decision. Collection and systematic analysis of data from multiple centers may bring clarity to the course and management of childhood multiple sclerosis. PMID- 19810866 TI - Cyclic AMP response element-binding protein and depression. AB - Depression is one of the most common and most devastating psychiatric disorders. Although a variety of treatment strategies is available, a major problem in its therapy consists of the unpredictability of the drug response. Furthermore, most antidepressant drugs, which usually increase 5-HT and norepinephrine levels in the synaptic cleft, are likely to produce side effects. Therefore, the quest for new options in antidepressant treatment is urgent. A novel therapeutic approach beyond manipulating the neurotransmitter-receptor interaction consists of targeting signal transduction and gene expression pathways. One of the best investigated pathways is the cyclic AMP second messenger system which ultimately influences gene expression by activating the transcription factor cyclic AMP response element binding protein via phosphorylation. There is evidence that this cAMP-PKA-CREB system is disturbed in depression and that an increased cyclic AMP response element binding protein activity may result in an improved neural plasticity, which in turn could contribute to amelioration of the clinical symptoms of depression. PMID- 19810867 TI - Atypical antipsychotics in the treatment of schizophrenic patients. AB - Atypical antipsychotics show a broader spectrum of efficacy and fewer adverse effects than older/typical neuroleptics. Therefore, they are used as the first choice treatment for patients suffering from schizophrenia. Concerning the acute effects, no differences among the group of new antipsychotics have been demonstrated so far. Thus, the most important criteria for choosing an antipsychotic for an individual patient are the expected side effects resulting from different receptor-binding properties. Today, no new antipsychotic can be considered as first-choice treatment for all patients. Outcome criteria should not only include psychopathology, relapse and rehospitalization, but also side effects, neuropsychological parameters and compliance, as well as self-rated well being and quality of life. PMID- 19810868 TI - Atypical antipsychotics in the therapy of bipolar disorders: efficacy and safety. AB - Bipolar disorders are characterized by recurrent manic and depressive phases with early onset and poor prognosis for chronic symptoms and premature death. Almost a third of patients do not improve with the available treatments. Atypical antipsychotics have been tested in the treatment of bipolar disorder as an alternative to typical neuroleptics due to their safer side effect profile and have been hypothesized to be mood stabilizers on the basis of their activity on affective symptoms of schizophrenia. The most significant studies published up to now, concerning the use of clozapine, olanzapine, risperidone, quetiapine and ziprasidone in bipolar disorders are reviewed here. The receptor profile of these drugs is related to their side effects. Finally, the current indications of atypical antipsychotics and future research are discussed. PMID- 19810869 TI - Virtual reality: a new tool for panic disorder therapy. AB - The use of a multicomponent cognitive-behavioral treatment strategy for panic disorder with agoraphobia is actually one of the preferred therapeutical approach for this disturbance. This method involves a mixture of cognitive and behavioral techniques which are intended to help patients identify and modify their dysfunctional anxiety-related thoughts, beliefs and behavior. Emphasis is placed on reversing the maintaining factors identified in the cognitive and behavioral patterns. The treatment protocol includes exposure to the feared situation, interoceptive exposure and cognitive restructuring. The paper presents a treatment protocol for panic disorder and agoraphobia, named experiential cognitive therapy, that integrates the use of virtual reality in a multicomponent cognitive-behavioral treatment strategy. The goal of experiential-cognitive therapy is to decondition fear reactions, to modify misinterpretational cognition related to panic symptoms and to reduce anxiety symptoms. PMID- 19810870 TI - Stroke in systemic lupus erythematosus. AB - With improved control of the immunological phenomena of systemic lupus erythematosus, the epidemiology of this disease is changing. While both cardiac and neurological manifestations of systemic lupus erythematosus have been consistently described, there is increasing awareness of the need for general cardiovascular prevention in systemic lupus erythematosus. An excess of stroke is seen in systemic lupus erythematosus especially, in those cases with anticardiolipin antibodies. There is increasing evidence for the contributory role of other cardiovascular risk-factors, including hypertension, hyperlipidemia and inflammatory markers, to the epidemiology of stroke and recognition of the role of endothelial dysfunction may play in systemic lupus erythematosus. Therapeutic approaches currently rely on anticoagulation alone. This review suggest that the full panoply of stoke interventions, including antihypertensive therapy and lipid-lowering, should be applied in the case of stroke in systemic lupus erythematosus just as in atherosclerotic stroke. PMID- 19810871 TI - Treatment of epilepsies associated with typical absences. AB - This review concentrates on the treatment of typical absences, which are distinct generalized epileptic seizures occurring mainly in the context of idiopathic generalized epilepsies. As typical absences may occur in combination with other generalized seizures, often in a syndrome-related pattern and vary in terms of frequency and severity, a brief discussion of the various different epileptic syndromes with typical absences and some principles of differential diagnosis are necessary. The diagnostic role of the electroencephalogram and its usefulness in monitoring the response to treatment are also outlined. To better comprehend the usefulness of the first-choice antiabsence drugs, but also the risks from those that are contraindicated, a brief section on the neurobiology of the mechanisms underlying absence seizures is also included. PMID- 19810872 TI - Neuroprotection in Parkinson's disease: love story or mission impossible? AB - Parkinson's disease was the first neurodegenerative disease in which a deficient neurotransmitter (dopamine) was successfully replaced by a systemically administered drug (levodopa). However, chronic levodopa therapy is associated with the development of motor and psychiatric complications. Furthermore, the progressive course of the disease is not halted and levodopa-resistant symptoms, such as dementia or postural instability, eventually appear. Most of these problems are related to the progressive nature of Parkinson's disease. Therefore, stopping or slowing the progression of Parkinson's disease is one of the main therapeutic objectives. Since the discovery of1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced parkinsonism in the early 1980s, the possibility of protecting dopaminergic nigral neurons against the action of toxic insults has attracted the attention of the neurological community. Could Parkinson's disease also be the first neurodegenerative condition with a potential neuroprotective treatment? The promise of rational neuroprotective therapy is generating much excitement and some frustration among researchers. In this article, the state of the art of neuroprotection in Parkinson's disease is reviewed. PMID- 19810873 TI - Therapy of neurodegenerative diseases using neurotrophic factors: cell biological perspective. AB - Neurotrophic growth factors are a structurally diverse group of endogenous proteins, which profoundly regulate survival and multiple other cellular functions of healthy and diseased neurons. Their physiological targets include all neuronal cell types affected by neurodegenerative diseases. Initial clinical trials, which were based on strong and convincing neuroprotective effects in animal models of neurodegenerative diseases, have failed so far. In this review, we adopt a cell biological perspective to outline both the potential and problems of the therapeutic use of trophic factors in the nervous system. Emphasis is given to the functional interplay of neurotrophic factors in the regulation of cell death, as well as to the effects these proteins exert on neuronal morphology and synaptic transmission. PMID- 19810874 TI - Possible role of nondopaminergic drugs on levodopa-induced dyskinesias in Parkinson's disease. AB - Levodopa remains the gold standard for the treatment of Parkinson's disease. However, the long-term use of this drug often leads to the development of motor fluctuations and dyskinesias. The main strategies for the prevention and therapy of levodopa-induced dyskinesias are related to modifications of levodopa administration or the early use of dopamine agonists. However, recent evidence from experimental models of parkinsonism and a number of clinical studies suggest that some nondopaminergic drugs could be useful in the prophylaxis or pharmacotherapy of levodopa-induced dyskinesias. Therapeutic approaches based on nondopaminergic drugs rely on the alterations of some neurotransmitters or neuromodulators induced by nigrostriatal denervation. PMID- 19810875 TI - Gene therapy in Parkinson's disease. PMID- 19810877 TI - Rasagiline: an anti-Parkinson drug with neuroprotective activity. AB - Mitochondria are now recognized as potent integrators and co-ordinators of cell survival/death and apoptosis. Therefore, they are pharmacological targets for induction or correction of excessive cell death in human pathology, which includes cancer and the neurodegenerative disorders Parkinson's and Alzheimer's disease. One such agent is the anti-Parkinson drug, rasagiline, a novel neuroprotective-antiapoptotic second-generation potent irreversible selective inhibitor of monoamine oxidase B. It prevents the neurotoxin-initiated demise of mitochondria via modulation of cell survival/death Bcl-2 family proteins, Bcl-2 Bax, at the mitochondria permeability transition pore, which regulates voltage dependent anion channels. It may also induce long-term potentiation - a consequence of its protein kinase C-dependent mitogen-activated protein kinase pathway activation. Rasagiline, unlike selegiline, is devoid of sympathomimetic action and is not metabolized to neurotoxic methamphetamine, but to aminoindan. Its neuroprotective activity is attributed to the aminoindan and propargylamine moieties and structural resemblance to certain metabotrophic glutamate 1 receptor antagonists. Ragagiline has been chosen for neuroprotective studies by the National Institutes of Health as a possible disease-modifying agent. PMID- 19810878 TI - Levetiracetam: its use in partial-onset seizure. AB - Levetiracetam is one of the newer antiepileptic drugs released within the past decade and is indicated for use as adjunctive therapy in adults with partial onset seizures. It is a novel antiepileptic drug with a unique activity profile and is chemically unrelated to existing antiepileptic drugs. The mechanism of action of levetiracetam does not involve conventional modulation of any of the three main mechanisms underlying classical antiepileptic drug activity. Levetiracetam pharmacokinetics are linear and time invariant. In addition, levetiracetam has shown clinical efficacy in randomized controlled trials for treatment of partial-onset seizures. Levetiracetam also has favorable pharmacokinetic and safety profiles. The purpose of this article is to examine the drug profile of levetiracetam, based on information derived from its preclinical and clinical studies. Furthermore, this article seeks to highlight key characteristics of levetiracetam that clinicians should consider in order to deliver individualized care to patients with epilepsy. PMID- 19810879 TI - Topiramate in migraine prevention. AB - Migraine is a common episodic headache disorder characterized by attacks that consist of various combinations of headache and neurologic, gastrointestinal and autonomic symptoms. Migraine with aura may be due to neuronal hyperexcitability, perhaps from cortical disinhibition. It is likely that headache results from the activation of meningeal and blood vessel nociceptors combined with a change in central pain modulation. Antiepileptic drugs are increasingly being recommended for migraine prevention since placebo-controlled, double-blind trials prove them to be effective. Topiramate is a structurally unique antiepileptic drug that was discovered by serendipity. It is a derivative of the naturally occurring monosaccharide D-fructose and contains sulfamate functionality. Topiramate may bind to membrane channel complexes at phosphorylation sites in the inner loop and, thereby, allosterically modulate ionic conductance through the channels. The topiramate MIGR-001, MIGR-002 and MIGR-003 trials represent the largest controlled trials of a migraine preventive agent ever performed. Treatment with 100 or 200 mg/day of topiramate was associated with significant reductions in migraine frequency, migraine days and the number of migraine attacks per month. Topiramate was also associated with a reduced use of acute medications. The most common adverse events were difficulty with concentration and attention, difficulty with memory, mood problems, anorexia and weight loss, paresthesias, hypoesthesia and language problems. Topiramate is a first-line migraine preventive drug and should be considered preferentially for all patients in whom weight gain is a concern, who are currently overweight or who have coexistent epilepsy or bipolar disease. PMID- 19810880 TI - Botulinum toxin treatment of adult spasticity. AB - Spasticity is a common complication of CNS injury and a cause of considerable discomfort and disability for the patient and difficulty for caregivers. It is estimated that over half a million people in the USA are affected by spasticity. In recent years, advances in the treatment of spasticity include the use of intrathecal baclofen, addition of tizanidine to oral medication and the introduction of intramuscular botulinum toxin injections. This review aims to give an overview of one of those advances, the use of botulinum toxin. The term spasticity refers only to a type of increased muscle tone due to overactive stretch reflexes, but there are other forms of muscle overactivity that follow brain or spinal cord injury which cause problems. Generally, however, the clinical picture is dominated by the neurological deficits that result from CNS injury. A useful concept in understanding the motor consequences of injury to the CNS is that of the upper motor neuron syndrome. PMID- 19810881 TI - Sertraline for social anxiety disorder. AB - Social anxiety disorder is a common, chronic and often debilitating condition. Selective serotonin reuptake inhibitors have been widely used for the treatment of anxiety disorders, with more data currently available for paroxetine. With its beneficial side-effect profile and proven efficacy in social anxiety disorder, sertraline can be considered a valuable addition to the variety of treatments currently available for this disorder. PMID- 19810882 TI - Neuroprotective treatment for Parkinson's disease. AB - Parkinson's disease is a progressive disorder that may ultimately lead to severe disability. Its course may be slowed or arrested by neuroprotective interventions that influence the basic mechanisms involved in neuronal death. In the last 2 years, claims have been made for several possible neuroprotective treatments, and patients and physicians are increasingly enquiring about such a therapy. The present report summarizes the basis of various potential neuroprotective strategies and the progress that has been made in establishing their validity and clinical utility. PMID- 19810883 TI - Dopamine agonists and their role in Parkinson's disease treatment. AB - Dopamine agonists directly activate dopamine receptors, bypassing the presynaptic synthesis of dopamine. There are two main classes of dopamine receptors: the D1 class linked to the enzyme adenyl cyclase and the D2 coupled to G-proteins that inhibit adenyl cyclase. Dopamine agonists are effective as monotherapy in early Parkinson's disease and as an adjunctive treatment to levodopa in the advanced stages of the disease. Dopamine agonists are increasingly used early, particularly in young-onset Parkinson's disease due to their levodopa-sparing effects and putative role as neuroprotective agents. The potential neuroprotective property of agonists has been the focus of research and debate in recent years. This review summarizes the use of dopamine agonists in Parkinson's disease and reviews the laboratory, clinical and functional imaging evidence of neuroprotection in this class of drug and discusses the clinical implications of their use in Parkinson's disease management. PMID- 19810884 TI - Complications and adverse effects of deep brain stimulation in Parkinson's patients. AB - Deep brain stimulation of subcortical structures, such as the subthalamic nucleus, internal pallidum or ventral intermediate nucleus of the thalamus, are currently used for the treatment of advanced Parkinson's disease with motor fluctuations or during resistant tremor. This review discusses the various adverse events occurring with this surgery by focusing on each stage of the procedure and the various stimulated targets. Intraoperative complications are rare whichever technique or type of target is used. The most frequent postoperative side effects are observed during stimulation of the subthalamic nucleus. It is likely that the small size of the nucleus and the possible spreading of current to the surrounding functional areas contribute to this phenomenon. However, these side effects are generally mild and transient compared with the dramatic improvement in motor symptoms and fluctuations. Deep brain stimulation of the internal pallidum and thalamus are not effective on all the motor symptoms in Parkinson's disease but adverse effects appear less frequently. Whatever the target, intraoperative complications and secondary side effects are being reduced by the use of new neuroimaging technology associated with an accurate intraoperative neurophysiological investigation. PMID- 19810885 TI - Hippocampal sclerosis and the syndrome of medial temporal lobe epilepsy. AB - Medial temporal lobe epilepsy due to hippocampal sclerosis is the most common epileptic syndrome and, if medically refractory, is a progressive disorder. Advances over the past decade allow this clinicopathological syndrome to be diagnosed in vivo. Many patients with hippocampal sclerosis become refractory to antiepileptic medications and are at risk of progressive hippocampal damage, cognitive deterioration and other disabling manifestations of refractory epilepsy. Fortunately, if hippocampal sclerosis is detected early and treated surgically, most patients with this syndrome can be rendered seizure-free, thus sparing them from further progression and disability. PMID- 19810886 TI - Role of clobazam in the treatment of epilepsies. AB - The antiepileptic effect of clobazam was first described in 1973. The main advantages of clobazam as an antiepileptic drug are its fast-onset of action and effect against a wide variety of seizures. Despite sometimes losing its effectiveness after a previous period in which it has been effective, clobazam may keep its antiepileptic effect when used intermittently. Tolerance is much more common with benzodiazepines than the other antiepileptic drugs. Although tolerance to clobazam does occur, sustained responders have been identified, and many patients maintain their controlled seizures for several months. Clobazam is a safe drug, and its efficacy is probably equivalent to that of the new antiepileptic drugs. Therefore, it should be considered as add-on medication in the treatment of epilepsy. Clobazam is rapidly and completely absorbed and can be used intermittently, especially for catamenial epilepsy and in patients with clusters of seizures, but only during periods of seizure exacerbation. PMID- 19810887 TI - Rasmussen's encephalitis: update on pathogenesis and treatment. AB - Rasmussen's encephalitis is a devastating disease characterized by refractory epilepsy and progressive atrophy of one cerebral hemisphere. The only treatment option able to prevent seizures is the surgical disconnection of the affected hemisphere. The etiology of the disease remains unknown, but several laboratory findings have recently indicated the involvement of the immune system. Whether these findings represent an initiating event or the consequence of a not yet identified trigger factor must be elucidated. Nevertheless, these observations suggest the use of different immunomodulatory approaches (including corticosteroids, plasmapheresis, intravenous immunoglobulins and immunosuppressive drugs) that proved to be of variable efficacy in some patients, confirming the potential role of the immune system, at least in the perpetuation of the disease. PMID- 19810888 TI - Current and emerging treatment options for migraine and other primary headache disorders. AB - Primary headache disorders are highly prevalent worldwide. The impact of primary headaches to the individual is significant and reflects physical suffering and decreased social and occupational functioning. The economic burden to society is enormous and represents direct healthcare costs and the indirect costs associated with decreased workplace productivity and work absences. The last decade has witnessed tremendous advances both in our understanding of the biology of headache and in our therapeutic armamentarium. This review outlines how these developments may be rationally implemented by highlighting individual treatment options and general treatment strategies. The state-of-the-art methods for the abortive and prophylactic treatment of tension-type headache, migraine and cluster headache are reviewed. PMID- 19810889 TI - Therapeutic implications of the modular headache theory. AB - A theoretical approach to understanding the primary headaches not yet classified by the International Headache Society classification system has been developed by the authors. It is proposed that groups of neurons, called modules, become activated to produce each symptom of a primary headache disorder and these modules are linked together to produce a headache. Headaches develop phenotypic stability through the process of learned stereotypy. This theory explains the huge diversity of headache phenomenology. It has implications for the classification, research and treatment of headache patients. The modular headache theory has therapeutic implications by directing us to focus on treatable modules and avoiding unnecessary treatment for less treatable symptoms. This allows for rational approaches to CNS hyperexcitability and incorporates the temporal patterns of modular activation into the patient's treatment plan. PMID- 19810890 TI - Use of neural stem cells as therapeutic vehicles for the treatment of malignant glioma. AB - The prognosis for patients with malignant glioma, the most common primary intracranial neoplasm, remains dismal despite significant progress in neuro oncological therapies and technology. This stems from the inability of current treatment strategies to address the highly invasive nature of this disease. Malignant glial cells often disseminate throughout the brain, making it exceedingly difficult to target and treat all intracranial neoplastic foci with the result that tumor recurrence is inevitable despite aggressive surgery and adjuvant radio- and/or chemotherapy. The use of neural stem cells as delivery vehicles for tumor toxic molecules is a novel experimental strategy aimed specifically at targeting disseminated tumor pockets. It has been demonstrated that neural stem cells possess a robust tropism for infiltrating tumor cells and that they can be used to deliver therapeutic agents directly to tumor satellites with significant therapeutic benefit. The aim is to develop these findings into a clinically viable technology which would not be hindered by ethical and tissue rejection related concerns stemming from the use of fetal or embryonic tissue transplants. A novel technology whereby neural progenitors similar in morphology, phenotype and behavior to fetal neural stem cells can be isolated from adult bone marrow is also discussed. These technologies represent important progress in the development of a treatment strategy that can specifically target disseminated neoplastic pockets within the brain. However, despite encouraging results in preclinical models, there are significant impediments that must be overcome prior to clinical implementation of this strategy. The key among these is understanding the specific tropic mechanisms that govern neural stem cells migration towards tumors and refining the processes used to generate neural progenitors from adult bone marrow in a clinically implementable fashion. Despite these limitations, the use of neural stem cells for brain tumor therapy holds significant promise and may emerge as an important therapeutic modality for patients with malignant glioma. PMID- 19810891 TI - Chronic migraine and medication overuse: still a debate. PMID- 19810893 TI - Use of botulinum toxin type A in poststroke spasticity. AB - The use of botulinum toxin type A to relax the overactive muscles occurring after stroke can improve the quality of life for affected patients. Botulinum toxin is injected locally into tight muscles and allows the patient improved function, such as cleaning the hand or assisting with dressing. A large multicenter, double blind, placebo-controlled trial demonstrated improvement in tone and functional disability in those patients with poststroke spasticity of the hand and wrist treated with botulinum toxin. Benefits were maintained throughout the 12-week study. Side effects of the injections were mild and self-limited. Botulinum toxin type A injections for upper extremity spasticity are safe and efficacious, and should be considered as first choice in the treatment of this disorder. PMID- 19810894 TI - Pathological glial reactions in neurodegenerative disorders: prospects for future therapeutics. AB - Pathological activation of the immune-competent glial cells is an obligatory event in neurodegenerative diseases. The secondary recruitment of astrocytes, resulting from an upgraded microglial activation, represents a critical point. Reactive astrocytes have to give up physiologically important functions (control of extracellular homeostasis and of synaptic transmission) and build a synergistic alliance with microglia in promoting oxidative, excitotoxic and beta amyloid-induced neuronal damage. Growing understanding of the pathogenically relevant molecular signaling pathways opens new possibilities of pharmacological corrections at the second messenger level. Here, the respective know-how of endogenous modulators, such as adenosine, might be used. The aim should be a titration of the glia reaction in order to maintain supposed beneficial functions of reactive microglia and to prevent the dangerous involvement of astrocytes. PMID- 19810895 TI - Nummular headache: diagnosis and treatment. AB - Nummular headache (coin-shaped cephalgia) has an unusual distinct feature: it is characterized by mild-to-moderate pressure-like pain exclusively felt in a rounded or elliptical area typically 2-6 cm in diameter. Although any region of the head may be affected, the parietal area is the common localization of nummular headache. The pain remains confined to the same symptomatic area which does not change in shape or size with time. The pain is continuous but lancinating exacerbations lasting for several seconds or gradually increasing from 10 mins to 2 h may superimpose the baseline pain. The temporal pattern is either chronic or remitting. Pseudoremissions may be observed when the pain reaches a very low grade or only discomfort (not pain) in the affected area is reported. At times, discomfort may prevail. Either during symptomatic periods or interictally, the affected area may show a variable combination of hypoethesia, dysesthesia, paresthesia or tenderness. Physical and supplementary examinations are normal. Nummular headache emerges as a primary clear-cut clinical picture. The particular topography and signs of sensory dysfunction make it reasonable to vent the idea that nummular headache is an extracranial headache, probably stemming from epicranial tissues such as terminal branches of sensitive nerves. Nummular headache may seem to be the paradigm of epicranias (group of headaches and pericranial neuralgias stemming from epicranial tissues). Nummular headache must be distinguished from head pain secondary to local processes and from tender points of more extensive headaches. Although nummular headache may frequently coexist with other primary headaches, it has an independent course. Treatment is seldom necessary and in most cases simple reassurance is sufficient. PMID- 19810896 TI - Treatment of chronic overmedicated headache patients. AB - The effective treatment for chronic headache with analgesic overuse is examined in this review. The first step of treatment is how to stop daily use of the analgesic, supporting the withdrawal syndrome and the worsening headache. In this phase there are different strategies from no treatment to antimigraine drugs. This step is followed by preventive antimigraine treatment sometimes followed or accompanied by antidepressant drug treatment. The most widely used drug in this class is amitriptyline [Tryptizol, Merck & Co, NY, USA. However, newer selective serotonin reuptake inhibitor antidepressants are also proposed. Unfortunately, few good clinical practice studies with long-term follow-up are available. In conclusion, there are no definite guidelines to treat these headaches and probably the main aim of the therapy for these patients should be to enable them to feel in control of their migraine, rather than feeling that migraine or analgesic drugs control them. PMID- 19810897 TI - Genetic factors in cluster headache. AB - Genetic factors are likely to play a role in cluster headache but are not simply accounted for. A small number of family studies have illustrated the role of genetics, and a few inconclusive studies assessed candidate genes for cluster headache. These clinical genetic studies are reviewed and the author's results from a large database of cluster headache patients are reported. The importance of genetic factors in cluster headache pathophysiology and comment on difficulties in genetic research of cluster headache are discussed. PMID- 19810898 TI - Treatment of migrainous vertigo. AB - Migraine and balance disorders are becoming increasingly recognized as comorbid conditions. Diagnostic criteria have recently been developed to facilitate the identification of patients for whom balance complaints are likely to be symptoms of migraine. The significant impact that associated balance complaints add to migraine requires evaluation for specific treatment of balance disorder symptoms in migraineurs. Treatment of migraine-related balance symptoms includes both migraine medications and nonpharmacologic therapy. Vestibular rehabilitation is an effective nonpharmacologic therapy for migraine-related vertigo and is optimally effective in severely affected patients when combined with migraine preventive medications. PMID- 19810899 TI - Assessing disability using the migraine disability assessment questionnaire. AB - The World Health Organization defines disability as the consequences of illness on ability to work, and ability to function in work and nonwork activities. For migraine, disability is the major determinant of the cost of illness. The most frequently used disability instrument in headache research is the migraine disability assessment questionnaire. Migraine disability assessment consists of questions that focus on lost time in three domains: school work or work for pay; household work or chores; and family, social and leisure activities. All questions ask about either days of missed activity or days where productivity was reduced by at least half. The main attributes of the migraine disability assessment questionnaire are reviewed. The applications of the migraine disability assessment questionnaire in clinical practice as well as public health initiatives are also discussed. PMID- 19810900 TI - Cannabinoids for the treatment of multiple sclerosis: no smoke without fire? AB - This review evaluates the most recent evidence available on the potential of cannabinoid drugs to relieve spasticity and pain in multiple sclerosis. Many different drugs are used to control the symptoms of multiple sclerosis, periods of relapse and the progression of the disease but it is in the potential management of spasticity, pain and lower urinary tract symptoms that cannabinoids have excited interest. At present, there are relatively few published, controlled clinical studies that support the efficacy of cannabinoids in this context and the evidence that is available is contradictory. On the other hand, the results of unpublished clinical trials are more encouraging and there are other major clinical trials underway. The results of animal studies are also consistent with the hypothesis that endogenous cannabinoids and their receptors are involved in the generation of spasticity and pain. PMID- 19810901 TI - Appraisal of the multiple sclerosis functional composite. AB - Multiple sclerosis is a chronic disease with widely varying clinical manifestations and a clinical course that fluctuates over short periods of time. Ideal outcome measures for assessment of multiple sclerosis-related impairment have been lacking but are increasingly needed as new therapeutic agents are developed and active control groups are used in clinical trials. The multiple sclerosis functional composite was developed in an attempt to create a better outcome measure for multiple sclerosis and indeed it has many of the desired features of an ideal outcome measure. The methods used to develop and validate this outcome measure are discussed, as well as potential areas for improvement. PMID- 19810902 TI - Delirium: pathophysiology, recognition, prevention and treatment. AB - Delirium, a clinical syndrome characterized by the acute onset of confusion, indicates that there is a disruption in physiological equilibrium that may lead to death unless quick and appropriate action is taken. Despite its frequent appearance, delirium often goes unrecognized, undetected and untreated, leading to poorer outcomes including longer lengths of hospitalization, persistent deficits in cognitive function and the need for nursing home placement following acute treatment. Prevention and treatment strategies do not need to be complex or expensive, but require well co-ordinated interventions from multiple disciplines including nursing, psychiatry, neurology and primary care, as well as the co operation and significant effort of family and friends who know the affected individual well. PMID- 19810903 TI - Prospects for the treatment of stroke using gene therapy. AB - Recent advances have demonstrated the use of gene therapy in the treatment of stroke in experimental animal models of focal ischemia, global ischemia and subarachnoid hemorrhage. Several different vectors for gene transfer have been studied including herpes simplex virus, adenovirus, adeno-associated virus and liposomes. Genetically modified cell lines (e.g., bone marrow-derived cells) have been studied for ex vivo gene therapy. The effects of gene transfer to several brain regions including the striatum, cortex, hippocampus, subarachnoid space and blood vessels are reviewed. Targets of gene therapy, such as molecular cascades after ischemia onset (Ca2+ influx, ATP loss, increased nitric oxide) and events associated with apoptosis are also reviewed, in addition to how gene transfer may be used to understand pathomechanisms underlying ischemic injury and the temporal therapeutic windows following ischemia within which protective effects of gene therapy have been achieved. The prospects for gene therapy for stroke are discussed in light of these findings and it is concluded that solutions to key technological problems will allow gene therapy to be a viable treatment modality. PMID- 19810904 TI - Neuronal plasticity in poststroke aphasia: insights by quantitative electroencephalography. AB - After stroke, the interhemispheric reorganization of the neural network implicated in language is hypothesized to be a function not only at the site of the lesion but also of dynamic right hemispheric and subcortical neural systems. These neural systems may have different functional shares in the utilization of particular language tasks. Important insights in language rehabilitation have been gained by quantitative topographical electroencephalography. It has been demonstrated that abnormalities within and outside speech relevant areas are related to restitution of poststroke aphasia. In the ischemic regions they indicate local disturbances, outside they reflect failures in neuronal networks involved in the generation and propagation of the alpha-rhythm. Treatment by a neuromodulatory agent has shown a significant shift of alpha-rhythm from frontal to occipital regions which may be due to a restitution of corticothalamic circuits in parallel to an improvement of neuropsychological scores in different language domains. PMID- 19810905 TI - Natriuretic peptides and panic disorder: therapeutic prospects. AB - Natriuretic peptides differentially modulate endocrine and behavioral stress responses in preclinical and human studies. While atrial natriuretic peptide inhibits the hypothalamic-pituitary-adrenocortical axis, C-type natriuretic peptide exerts stimulatory activity. In rodents, atrial natriuretic peptide reduces anxiety, whereas C-type natriuretic peptide has anxiogenic effects (mediated via corticotropin-releasing hormone). Patients with panic disorder show lower basal ANP plasma levels but a more pronounced release during experimentally induced panic attacks compared with controls. This could explain the absent pituitary-adrenocortical activation during panic anxiety and its paroxysmal nature. Furthermore, the effects of the panicogen cholecystokinin-tetrapeptide are attenuated by ANP pretreatment in panic patients, while C-type natriuretic peptide demonstrates anxiogenic action in healthy humans. Atrial natriuretic peptide agonists and C-type natriuretic peptide antagonists may have potential as a new class of antipanic and anxiolytic psychotherapeutic medication. PMID- 19810909 TI - Increased patient autonomy through long-term antipsychotic delivery systems for the treatment of schizophrenia. PMID- 19810906 TI - Pharmacotherapy for refractory schizophrenia patients. AB - Most schizophrenic patients experience morbidity over the course of their illness, as the illness runs a chronic course and full remissions are infrequent. Therefore, defining treatment resistance among schizophrenia is problematic. Not all patients respond to antipsychotic medication treatment and an estimated 30 50% are considered resistant to treatment. Treatment resistance normally occurs along a continuum and most patients manifest varying degrees of resistance to antipsychotic medications. Essock and colleagues discovered that more than 60% of the patients in state hospitals met the criteria for clozapine therapy and, therefore, they may qualify for treatment resistance. PMID- 19810911 TI - Clomipramine use in obsessive-compulsive disorder. AB - The serotonin reuptake inhibitor, clomipramine and newer selective serotonin reuptake inhibitors have become recognized as the most effective monotherapy for obsessive-compulsive disorder. Meta-analyses have suggested that clomipramine may be superior to other selective serotonin reuptake inhibitors in treating obsessive-compulsive disorder. Double-blind, direct comparisons of the selective serotonin reuptake inhibitors and clomipramine, have demonstrated equal efficacy, fewer side effects and lower medication discontinuation rates in selective serotonin reuptake inhibitor patients. However, clomipramine continues to play a vital role in the pharmacotherapy of obsessive-compulsive disorder, being used when two or more selective serotonin reuptake inhibitor trials have not been sufficiently effective, which can occur in up to a third of patients. Recent investigation suggests potential roles for intravenous clomipramine and the combination of oral clomipramine with selective serotonin reuptake inhibitors and other medications in treatment-refractory obsessive-compulsive disorder patients. PMID- 19810912 TI - New advances in botulinum toxin therapy for pain. AB - Myofascial pain syndrome is a chronic pain syndrome that affects a focal or regional portion of the body, accompanied by manifestations of neuropathy. The main treatment goal is to desensitize supersensitive structures and restore motion and function, releasing muscle shortening and promoting healing. Therapeutic approach include MTP injections using botulinum toxin type A and stretch, treatment of psychological or behavioral abnormalities, physical therapy, electrical stimulation and massage. Spasticity is defined as a motor disorder characterized by a velocity-dependent increase in tonic stretch reflexes (muscle tone) with exaggerated tendon jerks resulting from hyperexcitability of the stretch reflex. This physiological events resulted in uncontrolled reflex activity (spasms) and increased muscle tone (rigidity). When used as part of an integrated antispasticity program, the dose of botulinum toxin type A may be adjusted to provide the precise degree of weakness needed to overcome spasticity, while preserving some strength for normal function. The benefits botulinum toxin type A can offer any particular patient depend on the location and degree of spasticity, but improvements in daily activities are usually obtained. In conclusion, botulinum toxin is currently an alternative to consider in the treatment of pain associated with myofascial pain syndrome and/or spasticity, based on a correct diagnosis and patient schedule program. PMID- 19810913 TI - Drug interactions in epilepsy care: perspective on the newer generation antiepileptic drugs. AB - Pharmacokinetic interactions involving the antiepileptic drugs have long been considered to be an unavoidable component of epilepsy treatment. Many of the 'older' generation of antiepileptic drugs, including carbamazepine, phenytoin and phenobarbital, are recognized to cause hepatic induction of drug-metabolizing enzyme systems, such as the cytochrome P450 and UDP-gluculronyltransferase. Such interactions are not uncommonly implicated in resulting in clinically significant treatment complications. During the latter half of 1990s, a number of new antiepileptic drugs have become available to clinicians. Generally speaking, a common feature of these 'newer' generation medications are improved pharmacokinetic characteristics, including an improved drug interaction profile. The aim of this review is to summarize the data, both experimental and clinical, regarding pharmacokinetic interactions with the newer antiepileptic drugs. PMID- 19810914 TI - Immunotherapy for epilepsy. AB - The study of antibodies to the glutamate AMPA receptor subtype 3 in the pathogenesis of severe epilepsy associated with Rasmussen's encephalitis helped to crystallize the concept of autoimmune epilepsy. This work has been used as a paradigm to investigate autoimmunity, especially the humoral components, in several severe childhood epilepsies and more recently, in some of the more common acquired adult epilepsies. In addition, it helped prompt many open-label trials of immunotherapy for various epilepsies. We discuss the evidence that antiglutamate AMPA receptor subtype 3 antibodies are pathogenic and review the findings of the studies of immunotherapy for epilepsy. We conclude that immune treatments can have useful disease-modifying effects in some rare epilepsies, such as Rasmussen's encephalitis, at least in the short term and that their potential should be studied in the management of some forms of severe adult epilepsy. PMID- 19810915 TI - Surgical management of intractable epilepsy associated with neuronal tumor. AB - Neuronal tumors, such as gangliocytoma/gangliogliomas and cerebral neurocytomas, are slow-growing, indolent tumors consiting of neoplastic neuronal cells. They are likely to be present with seizures, often medically intractable epilepsy. During surgery for neuronal tumors associated intractable epilepsy, an enlargement of a resection area beyond the tumor boundaries, with recordings of intraoperative and/or chronic electrocorticography, may improve the postoperative seizure outcome, since tumor surrounding areas may have 'cerebral microdysgenesis' and may thus be epileptogenic. In addition, when the mesiotemporal lobe structures are involved with the tumor on the neuroimaging and have epileptiform electrocorticography activities, tumor removal and additional removal of the hippocampus are recommended. PMID- 19810916 TI - Pharmacological approaches to disease-modifying therapies in Parkinson's disease. AB - Parkinson's disease is a movement disorder resulting from neurodegeneration of the basal ganglia. Parkinson's disease is usually diagnosed at approximately 55 60 years of age and affects approximately 1% of the population over 60 years. Dopaminergic cells located in the substantia nigra and whose terminals extend to the striatum degenerate slowly such that 60% of cells are already lost when clinical motor symptoms first become evident. In addition to the classic triad of Parkinson's disease symptoms, rest tremor, muscular rigidity and bradykinesia, abnormalities in postural reflexes, dementia and depression are important comorbid conditions. Current therapies are aimed primarily at replacing dopamine with the dopamine precursor L-dopa or by the use of direct acting dopamine receptor agonists. Adjunctive treatments with monoamine oxidase inhibitors, catechol-O-methyl transferase inhibitors and amantadine are also used. While providing very effective symptomatic therapy in early stages of the disease, these agents fail to halt disease progression. Thus, while these treatments generally provide excellent results for 2-5 years, quality of life for Parkinson's disease patients becomes increasingly poor 5-10 years after diagnosis. Symptoms that become increasingly problematic with disease progression include inconsistencies in motor control (response fluctuations), gait and balance abnormalities, cognitive loss, hypophonia and dysphagia. Therefore, in order to maintain an acceptable quality of life for patients with Parkinson's disease, therapies that provide not only symptomatic improvement, but also slow or stop disease progression are greatly needed. In this review, we will discuss possible mechanisms of cell death in Parkinson's disease and related potentially disease-modifying therapies. These therapies include dopaminergic cell tranplantation and the use of growth factors. Small molecules that may act as antioxidants, nicotinic receptor agonists, nitric oxide synthase inhibitors, immunophilins, excitatory amino acid-related (iGluR and mGluR agonists and antagonists) drugs and anti-inflammatory drugs will also be discussed. PMID- 19810917 TI - Managing the critical problems of advanced Parkinson's disease. AB - The symptoms of Parkinson's disease can become increasingly difficult to control as the disease advances, particularly with the development of motor complications, such as end-of-dose wearing-off and dyskinesias, following long term therapy. At this stage, the patient is frequently referred to a Parkinson's disease specialist for advice on their disease management. In this review, we provide an overview of the Parkinson's disease specialist's strategies for coping with such problems. This includes establishing the optimum and most rational L dopa treatment schedule, improving L-dopa absorption, use of catechol-O-methyl transferase inhibition, the addition of oral dopaminergic agonists and the use of subcutaneous injections or infusions of apomorphine or lisuride. The role of new surgical procedures to treat Parkinson's disease is also reviewed. Finally, we highlight the increasing importance of treatment strategies that stimulate dopamine receptors in a more continuous, less pulsatile manner as a way of reducing the risk of developing treatment-associated motor complications. PMID- 19810918 TI - SNRIs: mechanism of action and clinical features. AB - Specific serotonin and norepinephrine reuptake inhibitors are a relatively recent class of antidepressants which have specificities making them a therapeutic choice. They are characterized by a mixed action on both major neuroamines of depression: norepinephrine and serotonin. The double polarity of the reuptake inhibition of serotonin and norepinephrine ensures a profile of effectiveness comparable to tricyclic antidepressants and higher than selective serotonin reuptake inhibitors, especially in severe depression. The absence of affinity for muscarinic, histaminic and alpha1-adrenergic receptors and the absence of action on monoamine oxidase limits their adverse effects and allows them to be better tolerated than tricyclic antidepressants and similar to selective serotonin reuptake inhibitors. Currently, two drugs of this class are available, milnacipran and venlafaxine, but several more are in development. They are active on depressive symptoms, as well as on certain comorbid symptoms (anxiety, sleep disorders) frequently associated with depression. Serotonin and norepinephrine reuptake inhibitors allow an improved rate of response and a significant rate of remission, decreasing the risk of relapse and recurrence in the medium and long term. Due to their good tolerance, they can also be prescribed for long-term treatment and in high doses in refractory depression or with strong potential of relapse. For all these reasons, one can reasonably conclude that serotonin and norepinephrine reuptake inhibitors represent the therapeutic of choice in depression and that their prescription is likely to strongly increase in the years to come. PMID- 19810919 TI - Use of adrenocorticosteroid hormones in the treatment of multiple sclerosis. AB - Despite the fact that adrenocorticosteroid hormones have been used in the treatment of multiple sclerosis for over 50 years, the extent to which they affect clinical disease activity remains unclear. Their administration results in the modification of a host of immune functions, including the downregulation of inflammatory cytokines and adhesion molecules involved in the trafficking of lymphocytes and macrophages across the blood-brain barrier. This leads to a rapid suppression of inflammation in multiple sclerosis lesions and to the restoration of the integrity of the blood-brain barrier. Clinical studies of adrenocorticosteroids in the treatment of multiple sclerosis have demonstrated a more rapid resolution of relapse, but have not uniformly demonstrated an improvement in the extent of recovery. More recent studies suggest that adrenocorticosteroid hormones may bring about a dose-dependent decrease in MRI measures of disease activity lasting 6 months, delay the time to second relapse and delay the time to sustained progression in relapsing-remitting and secondary progressive multiple sclerosis. Though it is clear that adrenocorticosteroid hormones may have greater effects than previously realized, their role in the treatment of multiple sclerosis remains controversial because significant methodological differences between studies have often led to conflicting results. PMID- 19810920 TI - Assessment, pathophysiology and treatment of fatigue in multiple sclerosis. AB - This review will update current views of the physiopathology and treatment of fatigue in multiple sclerosis. Fatigue is a common symptom in multiple sclerosis, being reported by about a third of the patients. For many of them it is the most disabling symptom, with negative consequences on working activity and daily life. There are no objective measures of fatigue which is essentially based on subjective complaints. Even if fatigue may be influenced by motor disturbances and depression, it is largely independent from both. Peripheral mechanisms, such as muscular disuse and deconditioning, joint abnormalities and metabolic changes of muscular fibers, have very little role in multiple sclerosis fatigue. All the available data indicate that fatigue is a 'central' phenomenon, due to multiple causes. Neurophysiological studies revealed an impairment of volitional drive to the descending motor pathways and functional imaging studies fund a selective involvement of frontal cortex and basal ganglia. Therefore, a dysfunction of the circuits between thalamus, basal ganglia and frontal cortex, affected by the multiple sclerosis lesions and/or disturbed in their function by the products of inflammation could be the substrate of fatigue. No specific treatments are available - management strategies include medications, exercise and behavioral therapy - in most cases a combined approach is suitable. Enhancers of vigilance, like amantadine and modafinil, were shown to be effective in class I and II trials, however their effects are modest. Aminopyridines may indirectly influence fatigue by reducing nerve conduction block in motor fibers. Some recent studies suggest the positive effects of drugs on fatigue may be via reducing the inflammatory activity, such as for glatiramer acetate. PMID- 19810921 TI - Variants and differential diagnosis of Guillain-Barre syndrome. AB - Guillain-Barre syndrome is characterized by acute progressive weakness, areflexia and maximal motor disability that occur within 4 weeks of onset. Its clinical and pathological spectra range extends from classical acute inflammatory demyelinating polyneuropathy to axonal variants with (acute motor sensory axonal neuropathy) or without (acute motor axonal neuropathy) sensory involvement. Moreover, several clinical variants (Fisher syndrome, ataxic Guillain-Barre syndrome and pharyngeal-cervical-brachial weakness) have been proposed. The association of clinical characteristics of Guillain-Barre syndrome with specific infections and the presence of antiganglioside antibodies also has been reported. Guillain-Barre syndrome now is recognized as a heterogeneous disorder with various clinical manifestations and recent neurophysiological and pathological findings have led to its reclassification. This review summarizes current concepts about the clinical spectrum of Guillain-Barre syndrome and its variants and about differential diagnosis. PMID- 19810922 TI - Pathogenesis and treatment of human T-cell lymphotropic virus Type I-associated myelopathy. AB - Human T-cell lymphotropic virus Type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a chronic progressive inflammatory neurological disease. It is considered that virus-host immunological interactions are an important cause of this disease. Coexisting high HTLV-I proviral load and HTLV-I specific T-cells is an important feature of HAM/TSP. The immune response to HTLV I probably contributes to the inflammatory process of CNS lesions in HAM/TSP patients. Therefore, treatments for HAM/TSP aim to reduce HTLV-I proviral load and normalize the abnormal immune responses to HTLV-I. PMID- 19810924 TI - Acetylcholinesterase-inhibitors in the treatment of autistic disorders. PMID- 19810923 TI - Pathogenesis and treatment of paraneoplastic neurologic syndrome. AB - Paraneoplastic neurologic syndrome, a rare complication of carcinoma, includes various neurologic disorders, such as encephalomyelitis, paraneoplastic cerebellar degeneration, subacute sensory neuronopathy, retinal paraneoplastic syndrome, opsoclonus-myoclonus syndrome and stiff-person syndrome. Several antibodies to malignant tumor cells and neurons are detected in sera and cerebrospinal fluids of patients with this syndrome, however, there is no direct evidence of antiYo or antiHu antibodies' causative roles in neuronal loss. Recent studies showed cytotoxic T-cell activities against peptides of an antigen protein recognized by antibodies in the peripheral blood of patients with paraneoplastic cerebellar degeneration and antiYo antibodies, as well as in patients with antiHu syndrome. Treatment of paraneoplastic neurologic syndrome with plasmapheresis, immunosuppresive drugs, or intravenous immunoglobulin therapy has been attempted. Here, we discuss previous reports and theoretical treatments based on recent etiological hypothesis of paraneoplastic cerebellar degeneration. PMID- 19810926 TI - Treatment of depressive symptoms with quetiapine. AB - Depressive symptoms are a frequent component of schizophrenia and other psychotic illnesses. The treatment of psychoses with conventional (typical) antipsychotic agents may worsen depressive symptoms and many patients only partially respond to treatment. Typical antipsychotics are also associated with serious side effects, such as extrapyramidal symptoms, and sexual and menstrual dysfunction. Many of these pitfalls, however, can be avoided with atypical antipsychotics. Quetiapine, an atypical antipsychotic with proven efficacy in the treatment of psychotic symptoms in schizophrenia, also has efficacy for treating depressive symptoms in patients with schizophrenia and other psychiatric disorders. This suggests that quetiapine may also be effective in treating and preventing depressive symptoms in patients with affective disorders, such as bipolar disorder. A review of the evidence base supports the hypothesis that quetiapine does not cause treatment emergent depression and may even be useful in the treatment and prevention of depressive symptoms in patients with bipolar disorder. PMID- 19810927 TI - Mirtazapine in the treatment of mood and anxiety disorders. AB - Mirtazapine is a new antidepressant whose effects on presynaptic adrenergic receptors leads to increased serotonergic transmission, and thus its antidepressant and antianxiety effects. It is equal in practical effectiveness to any currently marked antidepressant but may exert its effects earlier than some others. It is safe, well-tolerated and a useful addition to the drugs currently available for the treatment of mood and anxiety disorders. PMID- 19810928 TI - Risperidone long-acting injection. AB - Risperidone long-acting is the first atypical antipsychotic medication available in an injectable sustained-action formulation, potentially conferring the advantages of an atypical antipsychotic together with assured medication delivery. In addition, unlike depots of conventional antipsychotics composed of a prodrug in a viscous oil vehicle, this formulation uses microsphere technology to encapsulate risperidone in a biodegradable polymer. Progressive hydrolysis of the microspheres results in gradual release of risperidone leading to predictable and sustained plasma levels with repeated injections. PMID- 19810929 TI - Duloxetine: a review. AB - A developing concept is that antidepressant strategies which combine multiple mechanisms of action may have advantages over agents with single mechanisms. Duloxetine is a novel potent dual reuptake inhibitor of noradrenaline and serotonin. The antidepressant efficacy of duloxetine is reviewed in three trials. Results that emerge confirm the acute efficacy of the agent in major depressive disorder. In particular, remission rates in the comparative trials are higher with duloxetine than with either paroxetine or fluoxetine. Duloxetine appears to have specific efficacy in patients with somatic symptoms and there is some clinical evidence of analgesic properties. Modest weight loss was consistently described in the three acute trials and modest weight gain was seen in an open label follow-up study. Duloxetine was well-tolerated in all three trials with similar patterns of adverse events. No significant safety issues emerged from these trials. PMID- 19810930 TI - Recent genetic advances in ADHD and diagnostic and therapeutic prospects. AB - Attention deficit hyperactivity disorder (ADHD) is a common behavioral disorder of a complex nature. Genetic and environmental factors are thought to be involved in precipitating the disorder. Pharmacological, animal model and recent molecular studies support the role of genes (of minor or medium effect) from dopamine, serotonin and norepinephrine neurotransmitter systems in ADHD. Several investigations have pointed to the dopamine transporter, the dopamine receptors D4 and D5 and the serotonin transporter as genes of minor effect for ADHD. In addition, recent molecular analysis have also implicated synaptosomal-associated protein-25 and the serotonin receptor5-Hydroxtryptamine 1B as potential susceptibility loci for ADHD. An understanding of the genetics of ADHD will further facilitate refinement and validation of the ADHD diagnosis, and the development of reliable disease markers in the prediction of disease risk. A knowledge of genes that determine treatment response has the potential to be of predictive value and may also assist in rationalizing drug treatment in the ADHD population. However, increased understanding of inheritance brings its own challenges in the interpretation of new knowledge and in its wise and ethical use, especially in relation to future screening of at-risk individuals. This article attempts to review recent genetic advances and their possible implications for improved diagnosis and treatment. PMID- 19810931 TI - Preschool ADHD: exploring uncertainties in diagnostic validity and utility, and treatment efficacy and safety. AB - The current scientific and clinical status of preschool attention deficit hyperactivity disorder and its management is reviewed. Recent clinical and neuropsychological research supporting the utility of the construct will be presented along with a critical analysis of diagnostic issues. The published literature on treatment efficacy (both pharmacological and nonpharmacological) will be reviewed with a special focus on the issue of the safety and side effects of psycho-stimulants. The need for early identification and preventative intervention is indicated but caution should be employed in the use of psychostimulants with this age group. PMID- 19810932 TI - Fetal alcohol spectrum disorder and ADHD: diagnostic implications and therapeutic consequences. AB - The first association between prenatal alcohol exposure and a teratogenic effect in the developing fetus was made in 1968 by pediatrician Paul Lemoine in France. A few years later, Jones and Smith defined the fetal alcohol syndrome which combined dysmorphic facial, growth and CNS features. The classic fetal alcohol syndrome has been recently reviewed. Current research is beginning to acknowledge that exposure to differing levels of prenatal alcohol cause a spectrum of clinical conditions which can be grouped under the umbrella term fetal alcohol spectrum disorder. There are three subtypes to fetal alcohol spectrum disorder: full fetal alcohol syndrome, partial fetal alcohol syndrome (where there is only partial facial dysmorphology) and alcohol-related neurodevelopmental disorder (where there is no facial dysmorphology and often absence of growth features). The term alcohol-related neurodevelopmental disorder is gradually replacing the older term fetal alcohol effects. These subtypes were initially described by Stratton and colleagues in 1996. It is also becoming clearer that the full fetal alcohol syndrome as originally described by Jones and Smith, and diagnosed in dysmorphic clinics, is the least common clinical subtype resulting from prenatal alcohol exposure. In addition, current and ongoing research on brain structure, shape and function indicates that the brain abnormality is not correlated with the facial dysmorphology or intelligence quotient. Therefore, the presence of full fetal alcohol syndrome does not necessarily mean that the patient has the most severe CNS impairments. Genetic research has suggested that genetic factors may have a role in a modulating a woman's susceptibility to having a baby with fetal alcohol spectrum disorder (specifically the full fetal alcohol syndrome subtype). Thus, genetic factors may increase the variability of the teratogenic effect of prenatal exposure to alcohol on the developing fetus. PMID- 19810933 TI - Evaluation of radiotherapy for pediatric CNS tumors. AB - Despite the recent progress in neurosurgery and development of new chemotherapy drugs, radiotherapy is still an essential method of combined treatment for pediatric central nervous system tumors. The new approach of radiotherapy, such as conformal and stereotactic methods have recently been developed. These new methods and recommendations for treatment of pediatric brain tumors, such as astrocytoma, medulloblastoma, and ependymoma are presented. The side effects of treatment are also considered. In addition, the perspective of future development of radiotherapy in central nervous system tumors is presented. PMID- 19810934 TI - Neuromuscular symptoms in cancer patients. AB - The neuromuscular system is often affected in patients with cancer. This can be due to a direct effect of cancer, neurotoxic treatment, radiation therapy or paraneoplastic phenomena. This review focuses on the sites of involvement, such as cranial nerves, nerve roots, and plexus, mono- and polyneuropathies disorders at neuromuscular transmission, and muscle disease. At each site of the peripheral nervous system, the most common types of involvement and therapeutic issues are discussed. Knowledge of neuromuscular symptoms is important for medical oncologists, neurologists and neuro-oncologist alike, as precise diagnosis may have therapeutic consequences for the patients. Immunologic therapies, or in some cases anticancer treatment may decrease the individual's burden of disease in paraneoplastic syndromes. Conventional cancer therapies, such as chemotherapy and radiation therapy, also have neurotoxic side effects which need to be outweighed against their therapeutic effect. Several other less well-defined complications such as infection, hemorrhage or metabolic disorders have to be considered as a differential diagnosis. Complications of surgical and other interventions will not be discussed in this context. PMID- 19810935 TI - Immunotherapy of glioblastoma multiforme. AB - Glioblastoma multiforme is immunogenic and several glioblastoma multiforme related antigens have now been identified. In addition, the immunologic characteristics of the tumor microenvironment that may affect tumor growth are becoming increasingly understood. The type of immune-based approach selected to treat glioblastoma multiforme will depend on the tumor burden. For minimal disease states, active vaccination may be useful for generating adequate protection from relapse. However, for more advanced stage disease states, more rigorous strategies may need to be applied, such as adoptive T-cell therapy, antibody therapy or a combination of different techniques. The immunosuppressive environment observed during advanced malignancy may need to be reversed for improved efficacy of immune-based therapies. PMID- 19810936 TI - Molecular genetic alterations in malignant peripheral nerve sheath tumors. AB - Malignant peripheral nerve sheath tumor is a highly aggressive soft tissue tumor which is derived from neoplastic Schwann cells in most cases. Although sporadic malignant peripheral nerve sheath tumors are infrequent, their occurrence is considerably raised in the setting of neurofibromatosis Type 1. Allelic losses at the neurofibromatosis Type 1 locus on chromosome 17q are the best studied genetic alterations underlying malignant peripheral nerve sheath tumor so far. However, recent studies have suggested that alterations in cell cycle regulators, such as p53, are also involved in malignant peripheral nerve sheath tumor tumorigenesis. This review will focus on new findings about the molecular genetic alterations in both sporadic and neurofibromatosis Type 1-related malignant peripheral nerve sheath tumors, and will discuss their impact on further diagnostic advances, as well as on the clinical course of this tumor. PMID- 19810937 TI - Local delivery of antineoplastic agents using biodegradable polymers for the treatment of malignant brain tumors. AB - The prognosis for patients diagnosed with malignant brain tumors has remained dismal despite advances in both neuroimaging and conventional treatment modalities. The use of biodegradable polymers for controlled local delivery of antineoplastic agents represents a major advance in the treatment of brain tumors. By implanting polymers loaded with chemotherapy agents directly onto the brain tumor resection bed, therapeutic doses of a drug can be administered intracranially for prolonged periods of time meaning high systemic doses associated with debilitating toxicities can be avoided. This technological advance has expanded the spectrum of available treatments for neoplasms of the CNS and has facilitated new approaches for the treatment of malignant brain tumors. PMID- 19810938 TI - Liver transplantation as treatment for neurological disorders. AB - Liver transplantation has been performed for patients with incurable liver diseases but this therapeutic technique can be extended to neurological disorders, such as familial amyloid polyneuropathy and adult-onset type II citrullinemia. An amyloid precursor in familial amyloid polyneuropathy, a variant form of transthyretin, is produced mainly in the liver. After liver transplantation, it has been shown that this amyloid precursor disappears from the sera of patients once progression of the disease has halted. Type II citrullinemia is caused by a deficiency of liver-specific argininosuccinate synthetase activity and shows various neurological manifestations closely resembling those of hepatic encephalopathy, resulting in fatal outcome. All metabolic abnormalities in this disease, which include elevated plasma concentrations of citrulline and ammonia, can be immediately corrected by liver transplantation. PMID- 19810939 TI - Challenges in the pharmacotherapy of the epilepsies. PMID- 19810941 TI - Modafinil: past, present and future. AB - Modafinil is a novel wake-promoting agent approved for the treatment of excessive sleepiness associated with narcolepsy that holds significant promise as an alternative treatment to traditional psychostimulants for excessive fatigue associated with medical and psychiatric disorders and as augmentation medication for treatment-resistant depression. PMID- 19810942 TI - Heparanase: a target for therapy of brain invasive tumors? AB - Mechanisms responsible for the progression of malignant melanoma to highly aggressive brain-metastatic disease remain largely unknown. Brain neurotrophins can modulate the brain invasion of melanoma cells and the activity of an enzyme called heparanase. Heparanase is an endo-beta-D-glucuronidase that degrades the heparan sulfate chains of heparan sulfate proteoglycans, essential and ubiquitous macromolecules associated with the cell surface and the extracellular matrix of a wide range of cells and tissues. Human heparanase has been recently cloned as a single gene family and found to be a potential target for anticancer drugs because of its critical roles in angiogenic and invasive processes. The potential relevance of heparanase, as a cellular switch from noninvasive to the invasive phenotype and novel therapeutic target for invasive brain tumors, is introduced. PMID- 19810943 TI - Treatment of central neurocytomas. AB - Central neurocytomas are uncommon tumors of the CNS, representing approximately 0.1-0.5% of all primary CNS tumors. Patients most often present with symptoms of increased intracranial pressure (headache, nausea/vomiting, diplopia) due to obstructive hydrocephalus. Rarely, central neurocytomas may present with a hemorrhage. Central neurocytomas are intraventricular tumors with a predilection for arising in either the lateral or third ventricles. CT or MR cranial imaging demonstrates a circumscribed mass in the ventricles, frequent calcification and moderate contrast enhancement. Surgery provides definitive treatment, as little evidence exists as to response of these tumors to either radiotherapy or chemotherapy. Histopathology reveals a homogenous neoplastic cell population with neuronal differentiation, frequent calcification, occasional perivascular pseudorosettes and infrequent mitoses. Uncommonly, anaplastic variants of central neurocytomas (malignant central neurocytomas) are encountered and are distinguished by frequent mitoses, necrosis and endothelial cell proliferation. Following complete resection, central neurocytomas have a favorable prognosis usually obviating the need for either radiotherapy or chemotherapy. PMID- 19810944 TI - Epidemiology of pediatric tumors of the central nervous system. AB - Tumors of the CNS are the second most frequent malignancy in childhood and the most common of the solid tumors. An increase in CNS tumor rates during the past decades has been observed in a number of countries. One possibility is that improvements in diagnostics of CNS tumors contributed to the increasing trend. Another concern is that changes in environmental exposures may be responsible for the increasing incidence rates and a number of epidemiologic studies investigated factors like exposure to ionizing radiation, electromagnetic fields, or pesticides. In addition, prenatal exposures, for example from maternal diet, or the role of infectious agents were of particular interest. In this review, the current knowledge on causes of pediatric CNS tumors is summarized. PMID- 19810945 TI - Use of gamma knife radiosurgery for intracranial tumors. AB - Gamma knife radiosurgery of intracranial tumors is a noninvasive treatment with zero mortality and low and acute late morbidity. While microsurgery prevents growth of the tumor in the case of radical resection, radiosurgery prevents its growth, although the tumor is left in place. Contrary to microsurgery, the result of treatment using radiosurgery is not empty space in place of the tumor. In follow-up images, the tumor remains in place, but does not increase in volume. Eventually, regression of the tumor can be observed. Due to its epidemiology and incidence in the population, the most frequent tumor indications that lend themselves to gamma knife radiosurgery are meningiomas, acoustic neurinomas, pituitary adenomas and brain metastases. PMID- 19810946 TI - Molecular genetics of ADHD: prospects for novel therapies. AB - Attention deficit hyperactivity disorder has been shown to be a highly heritable disorder, leading to an increasing interest in genetic studies. While multiple genes may be involved, the candidate gene approach is based on postulated neurotransmitter mechanisms. Molecular genetic advances in relation to dopaminergic (dopamine transporter, dopamine D4 receptor and dopamine D5 receptor) genes, adrenergic, serotonergic and nicotinic receptor genes are reviewed. Comorbidity of attention deficit hyperactivity disorder with learning disability is discussed and possible genetic influences briefly reviewed. Recent pharmacogenomic studies of ADHD are reviewed and promising pathways suggested. Treatments 5 years from now may be more individually tailored in terms of gene/phenotype relationships. PMID- 19810947 TI - Recent advances in the pharmacotherapy of autism. AB - This review examines the most recent evidence for the pharmacologic management of severe maladaptive behaviors often observed in patients with autistic disorder. Although a multimodal approach is imperative in the management of autism, medications are frequently required for the individual to benefit from behavioral and educational interventions. Several classes of medications appear helpful in diminishing specific target symptoms and include the atypical antipsychotics, selective serotonin reuptake inhibitors, mirtazapine, mood stabilizers and alpha(2)-adrenergic agonists. Controlled research and longitudinal studies are needed to expand the knowledge base of these compounds as well as other medications under investigation. It is anticipated that future research regarding neuroimmunology, as well as the cholinergic and glutamatergic neurotransmitter systems, will contribute to our understanding of the pathophysiology of autistic disorder. PMID- 19810948 TI - Developmental pathways to severe antisocial behavior: interventions for youth with callous-unemotional traits. AB - There is an emerging body of research that has focused on understanding the different causal pathways through which children develop severe aggressive and antisocial behavior. In addition, there is a substantial body of research indicating that certain models of intervention have some demonstrated level of effectiveness for preventing and treating antisocial behavior, albeit with some significant limitations. The focus of this paper is to integrate these two bodies of research in an effort to improve the effectiveness of the next generation of interventions for antisocial youth, especially those designed to prevent and treat certain subgroups of antisocial youth who have largely been unresponsive to existing approaches to intervention. One such group consists of those antisocial youths who show a callous and unemotional interpersonal style and who seem to be at risk for showing a particularly severe and aggressive pattern of antisocial behavior. PMID- 19810949 TI - Efficacy of newer antidepressants for childhood anxiety disorders. AB - Anxiety disorders as a whole are the most common psychiatric disorders in children and adolescents, with a reported prevalence ranging from 6 to 18%. Childhood anxiety disorders predict adult disorders and the majority of adult anxiety disorders are antedated by childhood anxiety. Furthermore, social dysfunction may result from misdiagnosis of these early-onset disorders. Anxiety disorders can be managed using nonpharmacological and pharmacological options, or a combination of them. Different classes of medications have been used in pediatric anxiety disorders, including benzodiazepines, tricyclics and buspirone. Newer antidepressants (SSRIs and beyond) have fewer side effects, lower toxicity in overdose and a broader range of indications. This review will focus on the efficacy of these new drugs in the management of anxiety disorders in children and adolescents (obsessive-compulsive disorder, panic disorder, separation anxiety disorder/school refusal, generalized anxiety disorder, social phobia, post-traumatic stress disorder). PMID- 19810950 TI - Impact of smoking on anxiety disorders in adolescents. AB - Current literature suggests that cigarette smoking is associated with anxiety disorders, but the direction of the association between smoking and anxiety disorders has not been determined. This article provides an overview of the literature in the area of smoking and anxiety disorders in adolescents and current treatment strategies. PMID- 19810951 TI - Prospects for the use of cognitive-behavioral therapy in childhood obsessive compulsive disorder. AB - Obsessive-compulsive disorder is a common and debilitating anxiety disorder that is often seen in children and adolescents. In adults, cognitive-behavioral therapy has been demonstrated to be an efficacious and cost-effective treatment. Research with children is ongoing and preliminary results point toward cognitive behavioral therapy as an appropriate first-line treatment. Successful cognitive behavioral therapy with children requires not only skills in the application of behavioral techniques, such as exposure and ritual prevention, but also an understanding of how developmental issues impact treatment response and how to modify the therapeutic approach to match the child's needs. In this article, we review the outcome literature on cognitive-behavioral therapy for pediatric obsessive-compulsive disorder and discuss its successful application. PMID- 19810952 TI - Pharmacological treatment of pediatric obsessive-compulsive disorder. AB - Obsessive-compulsive disorder is a far from rare neuropsychiatric disorder in children and adolescents which affects approximately 1-1.5% of the pediatric population. Obsessive-compulsive disorder is defined by the presence of obsessions and/or compulsions. It often takes a chronic course with either persistent, constantly distressing obsessive-compulsive symptoms or a waxing and waning course. The best documented and most efficient treatment is cognitive behavioural therapy and antidepressant medication with serotonin specific agents. In most cases, an improvement rate of 20-50% is reported following medication alone. PMID- 19810953 TI - Current therapies for prophylaxis of pediatric migraine. AB - Migraine headaches are a frequent occurrence in the pediatric population. The prevalence in children and adolescents ranges from 3.2 to 10.6%. Frequent migraine attacks negatively impact on family dynamics and school attendance. Effective treatment strategies in adults with migraine are being developed through clinical trials. However, little information on the utility of current medication in pediatric migraine prophylaxis is available. With advent development of neurogenic hypothesis for migraine symptomatology, much recent interest involves the use of antiepileptic drugs for migraine prophylaxis. The ability to study these drugs in childhood migraine is made easier, especially since safety in children has already been addressed in pediatric epilepsy studies. PMID- 19810954 TI - Genetic prenatal and preimplantation diagnosis of trinucleotide repeat disorders. AB - This review examines the history, present status and future of genetic antenatal diagnosis for the trinucleotide repeat disorders, including Huntington's disease, Fragile X syndrome and myotonic dystrophy. Conventional prenatal diagnosis and the relatively new field of preimplantation genetic diagnosis, which can diagnose an affected embryo before a pregnancy is established, are described. Genetic diagnosis for these late onset diseases has inherent difficulties and many controversies. However, antenatal diagnosis is an important service for an individual with one of these mutations, who wishes to prevent the birth of an affected child. PMID- 19810955 TI - Cognitive therapy in bipolar disorder. AB - Stress-vulnerability models are increasingly viewed as plausible explanations of recurrence in severe affective disorders. This has promoted greater interest in the application of evidence-based psychological treatments, such as cognitive therapy, as an adjunct to medication for patients with bipolar disorder. This paper reviews the results from outcome studies of combined treatment approaches. Preliminary findings indicate that cognitive therapy reduces symptoms, enhances social adjustment and functioning and reduces relapses and hospitalizations in patients with bipolar disorder. However, the lack of published data from large scale randomized controlled trials and the absence of an adequate psychological model of manic relapse means that the role of cognitive therapy in bipolar disorders will be the subject of intense debate for some time to come. PMID- 19810956 TI - Preventing and treating Alzheimer's disease: strategies and prospects. PMID- 19810958 TI - Delivering safer services: can suicide and homicide among people in contact with mental health services be predicted? AB - Enquiries into incidents of violent deaths involving people in contact with mental health services have reinforced the view that such incidents can be predicted and prevented. However, concerns have been expressed that predicting those who will be involved in such incidents is impractical in clinical practice. Findings from studies that have attempted to develop methods for identifying those at greatest risk of violence are reviewed and the implications of the findings for efforts to develop safer mental health services are considered. PMID- 19810959 TI - Measuring the severity of chronic pain: a research perspective. AB - The authors of this review are members of the Aberdeen Pain Group, a group of multidisciplinary researchers who have been researching chronic pain for over 10 years. This review draws upon their experience to consider the measurement of chronic pain severity from a research perspective. The first half summarizes the requirements of a measure of pain severity for epidemiological research, describes a number of existing measures of pain severity and discusses the appropriateness of these instruments for measuring chronic pain as part of a postal epidemiological survey. The second half focuses on the use of the Chronic Pain Grade questionnaire by the Aberdeen Pain Group. The reliability and validity of this instrument as part of a postal questionnaire, its sensitivity to change over time, and how the use of such an instrument compares with retrospective perceptions of patients are all investigated. The review concludes with a brief discussion of future issues relating to the measurement of chronic pain severity, again from a research perspective. PMID- 19810960 TI - Neuromodulation for central pain. AB - Central pain, which follows brain and spinal cord injury remains an ill-treated entity affecting approximately 1-2 million people worldwide. When oral drugs fail, either immediately or in the long run, the only therapeutic option is neuromodulation, via either electrical or chemical means. The whole spectrum of neuromodulatory techniques for central pain is poorly appreciated by pain therapists. Results of all these techniques provide rational guidelines for the treating physician and are detailed in this review. Reviewed techniques include cortical, deep brain and spinal cord stimulation, transcutaneous electrical nerve stimulation, electroconvulsion, and pump infusion of intrathecal drugs. PMID- 19810961 TI - Acid-sensing ion channels and pain: therapeutic potential? AB - Many painful inflammatory and ischemic conditions such as rheumatoid arthritis, cardiac ischemia and exhausted skeletal muscles are accompanied by local tissue acidosis. In such acidotic states, extracellular protons provoke the pain by opening excitatory cation channels in nociceptors. From recent molecular investigations into sensory neurons, two important candidate genes (or gene families) encoding proton-activated cation channels - acid-sensing ion channel and transient receptor potential vanilloid receptor subtype-1 have emerged. Since both channels can be major pharmacological targets for pain therapy and therefore, for the development and screening of new analgesics, it is of great importance to confirm their involvement in acid-evoked pain in humans and to know their relative contribution to the nociception. The authors' recent psychophysical studies focused on these points and the data is presented and discussed in this review. PMID- 19810962 TI - Alzheimer's disease: from molecular pathogenesis to innovative therapies. AB - Alzheimer's disease is the most common neurodegenerative disorder and most prevalent cause of dementia in the elderly. Current standard treatment of Alzheimer's disease is based on the use of acetylcholinesterase inhibitors, which have shown symptomatic benefits on cognitive, functional and behavioral symptoms of the disease. A growing body of evidence suggests that the accumulation of amyloid beta-peptides may play a pivotal role in the pathogenesis of Alzheimer's disease (amyloid hypothesis). The incremental progress in elucidating the molecular basis of Alzheimer's disease is pointing the way towards more targeted and pathogenically specific treatment approaches. This review analyzes the available data on the new directions of Alzheimer's disease therapy, with particular focus on secretase inhibitors, amyloid beta-peptide vaccination,anti inflammatory agents, metal chelators and cholesterol-lowering drugs. PMID- 19810963 TI - Therapy of depression in dementia. AB - This is a comprehensive review of the existing diagnostic and treatment approaches to depression in Alzheimer's disease and related dementias. The existing evidence for the efficacy of pharmacological and nonpharmacological treatments are reviewed with an emphasis on the paucity of existing data and the need for more definitive controlled studies. In addition, data on the efficacy of such novel experimental treatments, such as various brain stimulation techniques (e.g., vagal nerve stimulation) are discussed. Alternative and psychosocial treatments are also reviewed. PMID- 19810964 TI - Therapies for neurologic erectile dysfunction. AB - A subset of men with erectile dysfunction have an underlying neurologic or neurodegenerative condition which directly causes their impairment. The authors briefly discuss the interplay of the CNS, peripheral nervous system and corporal bodies of the penis integral to normal erectile function and how these interactions are disrupted in the setting of various neurologic conditions. Effective therapies for these patients exist in a variety of different forms to include oral or injectable drugs and even surgery. Future treatment strategies will focus on the improvement of existing therapies as well as the invention of new long-term therapeutic options. PMID- 19810965 TI - Gender issues in multiple sclerosis. AB - Multiple sclerosis is an inflammatory demyelinating disorder of the CNS that also has features of a neurodegenerative disease. Gender influences both susceptibility to multiple sclerosis and the clinical course of disease. The basis for these differences may include genetic and immunological factors. The immunological differences between men and women may be mediated through the effects of the sex hormones. It can be speculated that there may also be gender differences in the ability of males and females to repair damage in the CNS. Multiple sclerosis is also influenced by pregnancy, with fewer relapses in pregnancy and increased numbers of relapses post-partum. This effect is likely to be mediated by the hormones produced in pregnancy, which include the steroid and other pregnancy-specific hormones. Pregnancy may also lead to a more favorable clinical course of multiple sclerosis. These observations lead to the possibility of treating multiple sclerosis patients with steroid or pregnancy hormones or the growth factors found in pregnancy. PMID- 19810966 TI - Therapy of trigeminal neuralgia secondary to multiple sclerosis. AB - Trigeminal neuralgia occurs in approximately 1-2% of patients suffering from multiple sclerosis. Its pathophysiology is common to idiopathic forms and resides in altered properties of the sensory axonal membrane at the root entry zone into the pons, leading to parossistic firing. Antiepileptic drugs of the sodium channel blocker type, such as carbamazepine, lamotrigine, and phenytoin are highly effective in controlling pain. However, side effects on the CNS may, at higher doses, severely worsen the already impaired neurologic conditions in multiple sclerosis patients. Baclofen, a presynaptic muscle relaxant is also beneficial in trigeminal pain. Whatever the drug, habituation and loss of efficacy are likely to occur sooner or later. Symptomatic, neurolesive surgery is indicated in cases resistant or intolerant to medical therapy. Radiofrequency thermorhizotomy, either monitored by trigeminal evoked potentials or not, is the recommended procedure, as it may be considered the most reliable as far as localization and degree of lesion are concerned. PMID- 19810967 TI - Current therapeutic approaches to primary CNS lymphoma. AB - The best therapeutic management in primary CNS lymphomas remains to be defined due to the small size and short follow-up of retrospective series, methodological pitfalls and limited number of prospective studies, and the paucity of randomized trials. This review focuses on the current therapeutic approaches, most commonly used drugs, role of intrathecal chemotherapy and indications for consolidation radiotherapy, providing recommendations for ordinary clinical practice. Some important therapeutic issues, such as the management of meningeal and intraocular lymphomas, as well as the relevance of salvage therapy as a playground for evaluation of new drugs, are also analyzed. Finally, the main open questions, as well as current and expected investigation trends are discussed. PMID- 19810968 TI - Gene therapy for malignant brain tumors. AB - Malignant primary and metastatic brain tumors have remained fatal in spite of major advances in diagnostic tools and the improvement of conventional therapies. Recent discoveries in the molecular basis of the disease have allowed increased understanding of the events that lead to the development of brain tumors and have also brought a new spectrum of alternatives for treatment. By using gene therapy, brain tumors can be treated by targeting their fundamental molecular defects, delivering gene-drugs to the malignant cells. The possible targets for this type of treatment are progressively increasing but abundant clinical success has yet to be obtained, in part due to imperfect delivery systems. In this review, the genetic fundamentals of various cerebral neoplasms and neurogenetic syndromes, different strategies used for gene therapy, various available DNA delivery systems, status of ongoing clinical trials, and possible prospects for the future are discussed. PMID- 19810969 TI - Clinical diagnosis of subcortical cerebral infarction. AB - Clinical diagnosis of subcortical infarction, chiefly lacunar stroke, has been considered important for arriving at the most rational stroke diagnosis and treatment strategy. This review focuses on the lacunar hypothesis and potential inaccuracy of clinical diagnosis and computed tomography in subcortical stroke. Modern magnetic resonance imaging techniques have increased sensitivity and specificity for subcortical infarcts and may help in determining the pathogenesis in individual patients. Full investigation is now favored for all stroke patients regardless of the clinical diagnosis. Acute stroke and secondary prevention treatments are discussed in reference to a diagnosis of subcortical infarction. PMID- 19810972 TI - Does gender play a role in Alzheimer therapy? PMID- 19810970 TI - Cluster headache mechanism and treatment. AB - There have been remarkable strides in the last decade in unraveling the mystery of primary headache disorders, such as migraine and cluster headache. The vascular theory has been superseded by the neurovascular phenomena which seems to be the permissive triggering factor in migraine and cluster headache. This has been achieved through new imaging modalities, such as positron imaging tomography and functional magnetic resonance imaging. Prior to these imaging techniques, it was impossible to study primary headache disorders since they have no structural basis. There is now an increasing body of evidence for the brain to be involved primarily in cluster and migraine and the vessel dilatation as an epiphenomenon. There are also better strategies for treating cluster headache. Most of the targets for abortive treatment remain vascular, however, for prevention neuromodulators, such as antiepileptics may be used in addition to calcium channel blockers. This review will discuss the treatment strategies based upon the pathophysiological basis for this disorder. PMID- 19810974 TI - Amantadine treatment for cocaine-dependent patients with severe withdrawal symptoms. AB - Despite a tremendous research effort, there are no medications currently approved for the treatment of cocaine dependence. Cocaine-dependent patients vary widely in clinical characteristics and response to treatment. This heterogeneity may partially account for the inability to find an effective medication. The identification of specific traits that predict response to particular therapies would be extremely useful in the search for a medication. Cocaine withdrawal symptom severity may predict response to medications that relieve cocaine withdrawal symptoms, such as amantadine. This paper describes a method of measuring cocaine withdrawal symptom severity and presents preliminary evidence demonstrating that amantadine may assist patients with more severe cocaine withdrawal symptoms achieve abstinence from cocaine. PMID- 19810975 TI - Advances in the therapy of high-grade glioma at relapse: pegylated liposomal doxorubicin. AB - Approaches to improve the prognosis of patients with high-grade glioma, which is still grim despite combined therapy and major advances in the development of new drugs, are highly important. Chemotherapeutic agents have not consistently produced favorable results in the relapse setting so far, with tumor responses reported in a minority of cases. Among them, doxorubicin has not shown significant efficacy, despite being one of the most effective substances in vitro and in animal models. Nevertheless, encapsulation of doxorubicin using polyethylene-glycol significantly improves penetration across the blood-brain barrier. Moreover, recent clinical trials in other entities have demonstrated that similar clinical responses can be achieved using approximately half of the dose of polyethylene-glycol-liposomes compared with conventional liposomes. Considering these facts, polyethylene-glycol-liposomal doxorubicin with and without tamoxifen was evaluated within two sequential Phase II trials performed at our institution. Polyethylene-glycol-liposomal doxorubicin (Caelyx) was efficient in a reasonable number of patients with equivalent results in comparison with other successful Phase II studies. This article will discuss the literature and our own results on polyethylene-glycol-liposomal doxorubicin, with a special focus on toxicity and efficacy data raised in clinical trials on patients with high-grade glioma. PMID- 19810976 TI - Nefazodone in the treatment of patients with post-traumatic stress disorder. AB - Post-traumatic stress disorder occurs in patients who have undergone a traumatic experience and manifests itself through a cluster of symptoms, including re experiencing, avoidance and hyperarousal. Post-traumatic stress disorder is commonly found among veterans of war and victims of sexual trauma, natural disasters and accidents. Nefazodone is a medication that has an FDA-approved indication for treating depression. Nefazodone has also been reported to be efficacious in treating post-traumatic stress disorder. Despite recent reports of hepatotoxicity, when used appropriately, nefazodone is generally as well tolerated as the medications currently FDA-indicated for post-traumatic stress disorder, the selective serotonin reuptake inhibitors. Through its mechanism inhibiting neuronal uptake of serotonin and norepinephrine and as a potent postsynaptic serotonergic antagonist, nefazodone has proven to be effective in treating post-traumatic stress disorder in several open-label trials. The results of such trials warrant its study in larger, double-blind, placebo-controlled clinical trials. PMID- 19810977 TI - Oral transmucosal fentanyl citrate: a novel agent for breakthrough pain related to cancer. AB - Pain is problematic for many cancer patients. As more experience has been gained in treating cancer pain, breakthrough pain has been recognized as a phenomenon that occurs frequently in this population. Breakthrough pain is generally treated with a variety of short-acting opioids via a number of routes. Actiq (oral transmucosal fentanyl citrate) is a formulation of fentanyl citrate delivered via the transmucosal route in a sucrose base. Patients using this agent are instructed to paint the unit on the buccal mucosa in order to achieve relief. The onset of Actiq is reported to be equal to intravenous morphine. There are a wide variety of dosage forms available, allowing 74% of patients involved in clinical studies of the agent to titrate to an appropriate dose. The proper use, side effect profile and advantages/disadvantages will be reviewed for this agent. PMID- 19810978 TI - Intrathecal steroid therapy for postherpetic neuralgia: a review. AB - Two recent publications advocated intrathecal injection of methylprednisolone acetate for postherpetic neuralgia. Conceptually, these therapeutic trials were based on the failed assumption that chronic inflammation persists in dorsal root ganglia even after the lesions of acute zoster have healed. Constructionally, these publications can be criticized on numerous grounds: cranial zoster was excluded, follow up periods were too short, dangers or reactivating the varicella virus were ignored, no methylprednisolone acetate-only group was included, pharmacological dangers of intrathecal lidocaine were overlooked. In addition to these basic conceptual deficiencies, multiple dangers of intrathecal methylprednisolone (therapeutically and by accidental injection) are manifest in a literature dating back over two decades in which several hundred serious complications are described. In summary, all rational evidence indicates that this treatment should not be prescribed. PMID- 19810979 TI - Genomics of immune molecules: early detection of cognitive decline and new therapeutic interventions. AB - Inflammation plays an important role in progressive degenerative diseases of the CNS, such as Alzheimer's disease, which represents the major cause of cognitive impairment in old age. Alzheimer's disease is a complex disease with multifactorial etiology and may represent a good model to investigate the multifaceted functions of immune responses in relation to cognitive decline. This review will discuss the following topics: relationships between inflammation and brain degenerative processes in Alzheimer's disease, the influence of genetic variations in cytokine genes upon the risk of developing Alzheimer's disease and/or the age at onset of the disease, the effects of gene allele polymorphism of immune molecules upon the turnover of these factors in the blood and brain of Alzheimer's disease patients and the association of genetic variations in some of this molecules with cognitive decline. PMID- 19810980 TI - Acetyl-l-carnitine as a possible therapy for Alzheimer's disease. AB - Alterations have been demonstrated in membrane phospholipid metabolism in Alzheimer's disease. Alterations in membrane phospholipid metabolite levels have been observed by 31P magnetic resonance spectroscopy 4 years before onset of cognitive changes. Implications of phospholipid membrane changes in Alzheimer's disease are presented. Augmentation of brain levels of acetyl-l-carnitine, an endogenous molecule, may help to reverse membrane phospholipid changes in Alzheimer's disease. Neurobiological effects of acetyl-l-carnitine and potential molecular mechanisms of acetyl-l-carnitine activity are reviewed. Clinical trials of acetyl-l-carnitine indicate that it is well-tolerated and may be effective in younger Alzheimer's disease patients and may be an effective adjuvant treatment. Acetyl-l-carnitine may especially benefit those Alzheimer's disease patients with depressive disorders. PMID- 19810981 TI - Pharmacotherapy of agitation and psychotic symptoms in Alzheimer's disease. AB - Behavioral disorders, such as agitation and psychosis, are the main reasons for nursing home placement of Alzheimer's disease patients. This review article discusses the efficacy and adverse effects rates of antipsychotics, the most effective and widely-used medications for psychosis in elderly patients and the most commonly prescribed class of psychotropic medicines in long-term care institutions. In addition, the efficacy of selective serotonin reuptake inhibitors is discussed. Recommendations are provided regarding these existing and potential new clinical strategies. A future perspective is provided, taking into consideration prospective developments in preventative treatments of Alzheimer's disease over the next 5 years. PMID- 19810982 TI - Neurosyphilis: is it still a clinically relevant form of dementia? AB - Though syphilis was thought by many to be a disease on the verge of distinction, a resurgence of cases since the 1980s has proven that this is not so. With this return of 'the great imitator,' a resurgence of neurosyphilis may soon be on its way. Relatively insensitive testing and more atypical presentations mean clinicians should continue to be vigilant for neurosyphilis. This article reviews the pathophysiology of syphilis and where neurosyphilis fits in the picture. It also reviews treatment options and recent guidelines submitted by the Center for Disease Control and Prevention. PMID- 19810983 TI - Bristol Activities of Daily Living Scale: a critical evaluation. AB - This article discusses the role of scales to measure activities of daily living, in particular in Alzheimer's disease, in the assessment and management of dementia. It explains the principles of activities of daily living measurement before introducing the reader to the Bristol Activities of Daily Living Scale. Many scales have been developed for measuring activities of daily living, but few were specifically designed for individuals with mild dementia living in the community for completion by caregivers. Fewer still were designed with the assistance of caregivers themselves. The article considers the use of the Bristol Activities of Daily Living Scale since its development and potential future changes. It also takes a longer look forwards at possible, more general advances in activities of daily living measurement. PMID- 19810984 TI - Transcriptional mechanisms underlying neuropathic pain: DREAM, transcription factors and future pain management? AB - Injury to peripheral nerves triggers a large-scale alteration in gene expression in sensory neurons as well as glial and immune cells. Neuropathic pain is believed to be the culmination of the plastic changes brought on by this 'genetic reprogramming'. However, an individual gene alteration may contribute to different processes induced by nerve injury, such as sensitization, regeneration or adaptation of endogenous analgesic mechanisms. Knowing which genes are altered in expression following nerve damage and perhaps, more importantly, understanding the transcription factors responsible for these changes, is a critical step toward more efficacious treatments for neuropathic pain. Recent genetic studies have revealed an important role for the novel transcriptional repressor, DREAM, in modulating acute and chronic pain and suggested the possibility of targeting this or other transcription factors for pain management in the future. PMID- 19810985 TI - New pharmacological strategies for pain relief. AB - Persistent or chronic pain is the primary reason people seek medical advice. Despite major advances in the neurobiology of pain, many patients with chronic pain still remain insufficiently relieved. The urgent medical need for novel and safe analgesics with high efficacy has led to intense research for new targets and we want to give a comprehensive overview on the current strategies in molecular pain research. The recently-discovered or re-evaluated targets that yielded compounds in clinical development will be summarized. In addition, we want to present emerging molecular strategies for pain relief, along with a mechanism-based classification of pain as the underlying concept for future diagnosis and therapy of chronic pain. PMID- 19810986 TI - Therapeutic options in neurosarcoidosis. AB - Neurosarcoidosis is an uncommon disorder that requires careful clinical evaluation to reach the diagnosis. It may involve the CNS and peripheral nervous system and in the majority of cases it is associated with systemic disease. The demonstration of the activity of the immune system, as suggested by routine and immunopathological studies, forms a theoretical basis for the common forms of immunosuppressive therapies that are employed in sarcoidosis. Classic treatment consists of steroids either intravenously or orally. In refractory cases or in order to reduce or eliminate steroids, methotrexate, cyclophosphamide, azathioprine, cyclosporin, chlorambucil, chloroquine and hydroxychloroquine use may be an option. PMID- 19810987 TI - Neurobehavioral and psychological changes induced by hyperthyroidism: diagnostic and therapeutic implications. AB - Clinical studies have demonstrated that neuropsychiatric symptoms may be common and often the only symptoms of patients affected by hyperthyroidism. The correct diagnosis of hyperthyroidism is crucial for optimal treatment because the neuropsychiatric symptoms tend to parallel resolution as the biochemical parameters of hyperthyroidism are normalized. For these reasons, a routine evaluation in every patient consulting for emotional pathologies is recommended, independent of other manifestations of thyroid disease. The effect of high levels of thyroid hormones in the CNS has been related to changes in the presence of alpha- and beta-adrenergic postsynaptic receptors, or more recently to changes in the serotonin concentration. However, the available information is incomplete and many questions remain to be answered in order to explain how the thyroid hormone modifies the CNS response. Increased knowledge of the molecular targets (i.e., genes) of thyroid hormones in the brain may help to provide an answer to these questions. PMID- 19810988 TI - Selective serotonin reuptake inhibitors in the treatment of generalized anxiety disorder. AB - Selective serotonin reuptake inhibitors have proven efficacy in the treatment of panic disorder, obsessive-compulsive disorder, post-traumatic stress disorder and social anxiety disorder. Accumulating data shows that selective serotonin reuptake inhibitor treatment can also be efficacious in patients with generalized anxiety disorder. This review summarizes the findings of randomized controlled trials of selective serotonin reuptake inhibitor treatment for generalized anxiety disorder, examines the strengths and weaknesses of other therapeutic approaches and considers potential new treatments for patients with this chronic and disabling anxiety disorder. PMID- 19810989 TI - Phenomenology and therapeutic options for dermatotillomania. AB - Dermatotillomania (also called psychogenic excoriation, neurotic excoriation, acne excoriee, skin picking and pathological or compulsive skin picking) is characterized by excessive scratching or picking of normal skin or skin with minor surface irregularities. Until recently, there have been few therapeutic options for patients with this disorder, which causes substantial distress and functional impairment in many patients. Recent pharmacological studies have found that some selective serotonin reuptake inhibitors may be effective in the treatment of this disorder. Furthermore, phenomenological studies have better characterized the heterogeneity of the behavior associated with dermatotillomania. Clinicians may now be able to develop more effective treatments for patients by targeting the specific behavioral symptoms of the disorder in individual patients. PMID- 19810990 TI - Cognitive dysfunction in multiple sclerosis: current approaches to clinical management. AB - In the last two decades clinicians have become increasingly aware of the frequency, importance and functional impact of cognitive dysfunction in multiple sclerosis. There is, however, a dearth of research in this area. In order to further progress in the field of neuropsychological diagnosis it is essential to validate sensitive, cost-effective and reliable screening instruments that can be used in clinical settings to identify patients appropriate for extensive neuropsychological assessment. As for the therapeutic approach, there is preliminary evidence that treatment with both disease-modifying and symptomatic agents may influence positively also the cognitive outcome of the patient. Few studies have assessed the efficacy of cognitive rehabilitation programs using an experimental design. Therefore, further research is clearly needed in this field: in particular, there is a need for tailor-made rehabilitation techniques for multiple sclerosis subjects, which take into account the course and stage of the disease, as well as the psychosocial problems of the individual patient. PMID- 19810991 TI - Treatment of myasthenia gravis: current practice and future directions. AB - Myasthenia gravis is the best understood of the autoimmune diseases and a number of treatments are currently used to produce clinical improvement. However, due to the scarcity of evidence-based and comparative data, there is still no consensus on many therapeutic issues. Even a widely accepted treatment like thymectomy has never been proven effective by a well-designed trial. These are just some of the unanswered questions: What is the best treatment algorithm and safest long-term management of myasthenia gravis? What patients are likely to benefit from thymectomy? How long should myasthenia gravis patients be treated? Is it possible to discontinue immunotherapy once remission has been achieved? What are the risks associated with long-term immunosuppression? In this article, we review current therapeutic strategies and these unresolved questions about myasthenia gravis treatment. PMID- 19810992 TI - Behavioral treatment strategies for migraine and tension-type headache: a review of the evidence and future directions. AB - Behavioral treatments have become widely accepted interventions for migraine and tension-type headache. Ample empirical evidence supports the effectiveness of interventions, such as relaxation training, biofeedback and cognitive-behavioral (i.e., stress-management) treatments. Such interventions may be utilized alone or in combination with the more widely used pharmacological treatments for headache. Behavioral approaches are often the treatments of choice for patients who have contraindications to or who are refractory to traditional pharmacologic interventions. Recent research has attempted to increase the availability of behavioral treatments to patients through more cost-effective forms of treatment administration. Ample empirical evidence now indicates that these efficient interventions are just as effective as more time-intensive, individually administered clinic-based treatments. Important future directions for behavioral headache treatments include: integration into the primary care setting, developing algorithms for matching behavioral and pharmacologic treatments to patient characteristics for maximum effectiveness, behavioral skills training to enhance outcomes with pharmacologic interventions and reduce complications (e.g., rebound migraine), as well as identifying and treating comorbid conditions that undermine treatment. PMID- 19810995 TI - Schizophrenia: a personal testament. PMID- 19810997 TI - Valproate: a practical review of its uses in neurological and psychiatric disorders. AB - Valproate is an antiepileptic drug with a wide spectrum efficacy in the management of various types of seizures and epileptic syndromes. In addition to its efficacy in epilepsy, valproate has been found to have therapeutic effects in various psychiatric disorders, including manic states, bipolar, panic and schizophreniform disorders and in the prevention and acute treatment of migraine. Valproate is available in various formulations that differ primarily in their absorption profiles. The purpose of this review article is to highlight the most important pharmacokinetic and pharmacodynamic properties of this drug that all clinicians should be aware of when using it as an antiepileptic, antimigraine, or mood-stabilizing agent. We review its use with other common antiepileptics or psychotropic drugs and the pharmacokinetic interactions with commonly used drugs. We also review its tolerance and toxicity profile in the various age-groups and devote a section to the particular problems associated with its use in patients with renal and hepatic failure and in women of gestational age. PMID- 19810998 TI - Adenosine as a neuroprotectant: therapeutic perspectives. AB - The potential for exploiting the neuroprotective properties of the purine nucleoside, adenosine, in a variety of CNS disorders, including: ischemic and traumatic injuries, neurodegenerative disorders, epilepsy and pain, has aroused considerable interest in both academic and pharmaceutical circles. A variety of approaches have been employed, ranging from the development of new selective agonists and antagonists for adenosine receptors, to compounds which can either potentiate extracellular levels of endogenously released adenosine or enhance its actions at receptors. Although many of these approaches were successful in animal studies, clinical trials have been delayed by the need to develop more potent and selective agents. With the recent promising advances in this area, future prospects for the development of new neurotherapeutic agents now appear promising. PMID- 19810999 TI - Risk, mechanisms and prevention of stroke after an acute myocardial infarction. AB - This review focuses on risk, mechanisms and prevention of stroke after an acute myocardial infarction. Pathophysiological aspects are covered and in particular, the relationship between ventricular thrombi and stroke is critically reviewed. New aspects concerning platelet activation and thrombotic risk is discussed. Epidemiological data are reviewed to form a background for a discussion concerning the absolute risk of stroke in different age-groups and the importance of different risk-factors. The common view that anterior location of an infarction is of major importance is challenged. Treatment options for prevention of ischemic, as well hemorrhagic, stroke is covered in detail. The author's opinion regarding practical clinical strategies, as well as more speculative comments about the future in the field, is included. PMID- 19811000 TI - Systemic thrombolytic therapy of acute ischemic stroke with rtPA. AB - Systemic thrombolysis with recombinant tissue plasminogen activator is the first therapy proven to be effective in acute ischemic stroke. Nevertheless, its efficacy and feasibility are still a matter of debate. Much of this debate derives from uncertainties regarding patient eligibility and problems implementing this effective, but potentially harmful therapeutic approach into daily stroke care. Whether a patient with acute ischemic stroke will receive thrombolytic treatment depends on several factors and prerequisites, of which time and expertise are most important. This review is focused on what is known and what remains to be understood in order to integrate thrombolysis into the routine treatment of a disease, which ranks third in mortality and first in acquired disability among adults in Western countries. PMID- 19811001 TI - New developments in pharmacotherapy of pediatric anxiety disorders. AB - Anxiety disorders affect about 10% of children and adolescents, are often chronic and impair psychological, academic and social functioning. Recent studies have clearly demonstrated the efficacy of selective serotonin reuptake inhibitors in young people with obsessive-compulsive disorder and mixed or generalized anxiety disorders. Evidence is emerging for effectiveness in social phobia and its variant, selective mutism. Since the effect size for selective serotonin reuptake inhibitors in anxiety greatly exceeds that in depression, studies with small numbers readily show superiority of medication over placebo. Hence, further positive studies are predicted in the next few years. As a result, the rate of prescription of selective serotonin reuptake inhibitors in young people has been rising and will continue to do so. These medications are well-tolerated, but motor and behavioral side effects are more common in young people. Long-term research and postmarketing surveillance are needed. PMID- 19811002 TI - Nonpharmacological therapies for trichotillomania. AB - Contrary to prior belief, trichotillomania is not a rare condition and may occasionally prove resistant to therapy. Although for practical reasons, pharmacological agents are used as first-line treatment, there are a variety of nonpharmacological therapeutic approaches available. In the pediatric age-group - in which the disorder is common - there is a strong tendency among clinicians and parents towards the avoidance of medications and the use of nonpharmacological strategies as the treatment of choice. This article reviews the multimodal approach to the trichotillomania patient and emphasizes the efficacy and safety of different behavioral, hypnotic, educational and family therapy techniques. Further controlled outcome studies are needed to substantiate this clinical impression. PMID- 19811003 TI - Monoamine and neuropeptide-related function: prospects for novel therapeutics of depression. AB - Treatment of depression has been dominated by monoamine hypotheses for nearly half a century. Although the ultimate downstream targets of manipulation of biogenic amine transport and metabolism are signal transduction cascades, neurotrophic factors and ultimately genomic fine-tuning, much of drug development has remained focused on strategies to facilitate a rapid initial augmentation of synaptic neurotransmitter levels. Both monoaminergic agents that are highly specific in their target receptor stimulation and agents that modify multiple monoamine systems will move through the stages of drug development in coming years. Furthermore, recent landmark successes and initial pilot data indicates that small molecule peptide receptor antagonists for substance P, corticotrophin releasing factor and eventually others will be of increasing importance, since they may more subtlety modulate neurotransmission and only act on already deranged neural circuits. Eventually treatment strategies may begin to target intracellular kinase and phosphatase activity and other signal systems responsible for transporter and receptor trafficking patterns, as well as via more direct pathways toward mobilization of neurotrophic factor activity. In the near future however, refinement of monoamine strategies to treat depression will continue to guide thinking and development of novel drugs for depression. This review focuses primarily on developments relevant for the evolution of monaminergic and neuropeptide-based drugs for depression. PMID- 19811004 TI - Management of melancholia. AB - There is increasing recognition that melancholic depression can be distinguished from other depressive types and that it differs from the nonmelancholic disorders in terms of its natural history, spontaneous remission rate, response to placebo and response to psychosocial and physical treatments. As it shows a preferential response to physical therapies, the psychotherapies are best viewed as adjunctive rather than primary treatments for melancholia. Of distinct importance, melancholia appears to show a differential response to differing antidepressant drugs, responding less well to the newer narrow-action antidepressant drug classes. For all these reasons, the management of melancholic depression needs to be distinguished from the management of 'depression' per se. PMID- 19811005 TI - Role of mood stabilizers in bipolar disorder. AB - Bipolar affective disorder is a common severe illness which is associated with a significant morbidity and mortality. The area of pharmacotherapy for bipolar disorders is an exciting one with a number of recent new developments. This article summarises the current state of evidence regarding the efficacy of lithium, valproate and carbamazepine for the treatment of bipolar disorders and also highlights the emerging role of the atypical antipsychotics and newer anticonvulsants. PMID- 19811006 TI - Chronobiology and insomnia: pathophysiology and treatment of circadian rhythm sleep disorders. AB - This review summarizes current knowledge of sleep disorders with a chronobiological basis, including: delayed sleep phase syndrome, advanced sleep phase syndrome, non24 h sleep-wake syndrome and irregular sleep-wake pattern disorder. These circadian rhythm sleep disorders are characterized by a misalignment between the timing of the sleep period with respect to the day-night cycle and as a consequence of patients attempting to maintain 'normal' social hours, reduced sleep quality. In addition to the specific circadian rhythm sleep disorders, this review will also examine current drug (e.g., hypnotics and melatonin) and nondrug (e.g., bright light therapy and chronotherapy) treatments, the overlap with psychophysiological insomnia and future directions. PMID- 19811007 TI - Inverse agonism at beta-adrenergic receptors: therapeutic implications. AB - Ligands for G-protein-coupled receptors constitute important means in therapeutic applications. Therefore, developing new ligands for these receptors has been one of the major therapeutic aim. In classical pharmacology, ligands for G-protein coupled receptors were classified as agonist, partial agonist and antagonist. Under this classification, it was believed that three different choices of ligands are available in the treatment of diseases or in the strategies to develop new drugs. Recently this classification has been extended to include a spectrum of (full) agonist, partial agonist, neutral antagonist, partial inverse agonist and (full) inverse agonist. The new classification has brought new perspectives in the understanding of pathophysiology, in the strategies of developing new drugs and in the treatment of diseases. Now the challenge is to figure out the appropriate choice of ligand for the treatment of relevant diseases. Inverse agonists became particularly interesting with the understanding that G-protein coupled receptors may have spontaneous activity that can be inhibited by these ligands. In this review, developments in the concept of inverse agonism at G-protein-coupled receptors and beta-adrenergic receptors and therapeutic implications of inverse agonists are discussed. PMID- 19811008 TI - Global impact of schizophrenia. PMID- 19811010 TI - Mental and behavioral dysfunction in movement disorders: an international symposium. PMID- 19811011 TI - 'Atypical' antipsychotics: where does ziprasidone fit? AB - Identified as the prototype of 'atypicality', clozapine heralded a new generation of antipsychotics intent on providing greater efficacy than 'typical' antipsychotics, while diminishing the risk of D2-related side effects such as extrapyramidal symptoms and hyperprolactinemia. Ziprasidone is the most recent in this new class of antipsychotics to enter the clinical market. PMID- 19811012 TI - Hormone treatment for mood disorders in women. AB - Periods of intense hormonal fluctuations have been associated with heightened prevalence and exacerbation of underlying psychiatric illness, particularly the occurrence of premenstrual dysphoria, puerperal depression and depressive symptoms during the menopausal transition. It has been speculated that sex steroids, such as estrogens, progestogens, testosterone and dehydroepiandrosterone, exert a significant modulation of brain functioning, possibly through interactions with various neurotransmitter systems. It is therefore intuitive that abrupt alterations of these hormones would interfere with mood and behavior. On the other hand, accumulating data suggest that hormonal interventions may also promote relief or even remission of depressive symptoms, as already demonstrated in studies on perimenopausal women treated with transdermal estradiol. This article reviews the potential role of sex hormones for the treatment of depressive disorders in women. There are preliminary, but promising data on the use of estradiol and testosterone for specific subpopulations. Further studies would help to better delineate the usage of these compounds as an antidepressant strategy (monotherapy or augmenting treatment). PMID- 19811013 TI - Factors affecting the use and selection of antidepressants in the elderly. AB - Depression is a common problem for the elderly. It is critical that clinicians become aware of this fact and that appropriate therapy is prescribed. Both pharmacotherapy and psychotherapy play roles in the treatment of depression in these patients. The appropriate use of medications requires an understanding of factors specific to the elderly that affect their selection and use of these medications. In particular, concurrent medical conditions, potential drug-drug interactions and pharmacokinetic changes need to be considered. The intent of this paper is to describe these factors and suggest some guidelines for the appropriate use of antidepressants in the elderly. PMID- 19811014 TI - Management of pediatric disruptive disorders. AB - Disruptive behavior disorders include conduct disorder and oppositional defiant disorder. Aggression, hostility, rule breaking, defiance of authority and violation of social norms are the primary behaviors seen in these disorders. There are no established fully effective psychosocial or behavioral therapies. This review of both behavioral and pharmacological approaches to treatment documents some degree of short-term benefit in research settings. Studies suggest a lack of significant long-term benefit to behavioral therapy when the conduct disorder is of any significant severity or when delivered in community settings. The long-term safety and effectiveness of pharmacotherapy of disruptive behavior disorder is not established. PMID- 19811015 TI - Prenatal risk factors and schizophrenia. AB - This article highlights major research findings that indicate a relation between prenatal environmental factors and the risk of adult schizophrenia. Evidence suggests that schizophrenia may have its origins in early maldevelopment of the brain. However, the factors responsible for this devastating disorder remain unknown. While genes clearly play a causal role, there is likely an environmental contribution as well. We discuss the evidence for several candidate environmental exposures that occur during prenatal life and that may modify the risk for developing schizophrenia. The main challenge of this type of research, termed risk-factor epidemiology, is to generate and test such exposures in subjects with schizophrenia, in order to develop preventive strategies and better understand the mechanisms that lead to this illness. PMID- 19811016 TI - New targets for antipsychotics. AB - Atypical antipsychotic drugs have a favorable side effect profile compared with older neuroleptics. Clozapine exhibits superior clinical efficacy in resistant schizophrenia. The neurochemical profile of these atypical agents suggests two distinct strategies in the development of novel antipsychotic drugs. Better targeted dopaminergic agents may avoid the side effects associated with profound D2 receptor blockade. Current approaches include D3 receptor antagonists and drugs combining partial agonist activity at dopamine autoreceptors with conventional D2 antagonism. The D1 receptor may be a potential target for improving cognitive function in schizophrenia. Secondly drugs acting at glutamate (NMDA) and serotonin (5-HT(2A), 5-HT(1A), 5-HT(2C), 5-HT(3)) receptors may offer improved treatment particularly against resistant schizophrenic symptoms, as well as improving the side effect profile of antipsychotic drugs. This research in receptor pharmacology is set in the context of the increasingly recognised importance of the clinical care for patients with first onset schizophrenia and recent developments in pharmacogenomics. PMID- 19811017 TI - Atypical antipsychotics: revolutionary or incremental advance? AB - The discovery of chlorpromazine half a century ago and the subsequent emergence of other first generation antipsychotics, heralded a new advance in the treatment of schizophrenia. However, these new medications were not always effective. Even when they reduced the positive symptoms of schizophrenia, they were not as helpful in the relief of other symptom domains of schizophrenia, such as negative symptoms, impaired cognition and persistent aggressivity. Clozapine was the first of the new second generation of antipsychotics. It was introduced in the USA specifically for the indication of treatment-refractory schizophrenia. However, clozapine's side effect burden has led to a search for its replacement. This quest has pointed out the limitations of our treatments for refractory patients, but has made available a variety of second generation antipsychotics that have raised our expectations. Furthermore, the atypical antipsychotics hold promise for the treatment of the nonpsychotic patient with mood dysregulation or acute agitation. PMID- 19811018 TI - Therapeutic potential of nervous system growth factors for neurodegenerative disease. AB - Fifty years after the discovery of nerve growth factor, the first gene therapy trial for the treatment of Alzheimer's disease utilizing nerve growth factor has begun. This review focuses on the history of nerve growth factor and the rationale behind targeting nerve growth factor therapy to rescue degenerating neurons of the cholinergic basal forebrain system in Alzheimer's disease. Further, the future steps necessary to turn the promise of gene therapy into a practical approach for treating Alzheimer's disease and other neurodegenerative disorders are discussed, including key issues, such as regulatable gene expression and making the transition from an ex vivo to an in vivo gene therapy approach. PMID- 19811019 TI - Management of Parkinson's disease. AB - Parkinson's disease is one of the most common and debilitating movement disorders. It affects a large number of the aging population and as the life expectancy of the average person increases, it will affect ever increasing numbers of people. Diagnostic accuracy remains problematic and treatment for some is unsatisfactory. New understanding of the mechanism behind neurodegeneration in this disease and the pathophysiology of the basal ganglia has helped in designing therapeutic paradigms. Patients diagnosed today have a far better quality of life than those diagnosed even 10 years ago, while patients diagnosed 10 years from now will fair even better. Research is advancing rapidly and will help to cure this disease in the next century. PMID- 19811020 TI - Adverse effects in the treatment of Parkinson's disease. AB - Side effects to antiparkinsonian drugs constitute an important component of the daily management of patients with Parkinson's disease. Treatment with levodopa frequently leads to motor fluctuations and dyskinesias. Hallucinosis, nausea and vomiting, drowsiness, orthostatic hypotension and peripheral edema can be managed by dose reduction, medication substitution and specific counteractive measures. Anticholinergics frequently cause cognitive or autonomic symptoms while ergot derived dopamine agonists carry unique, albeit rare, risks of fibrotic, vasoconstrictive and dermatological side effects. Current areas of controversy include: dopamine agonist-induced sleep attacks, increased mortality with the combination of selegiline and levodopa and the association of levodopa with melanoma. PMID- 19811021 TI - Pharmacological approaches to the treatment of excessive drinking and alcohol dependence. AB - Excessive drinking is common in Western societies and leads to a wide range of physical, psychological and social problems. These problems are costly to society. The 1-year prevalence of dependence is 4-5% and lifetime prevalence 13 16%. A wide range of pharmacological approaches to attenuate drinking or promote abstinence has been studied. Clinical research in this area has been affected by poor methodological quality, with some important exceptions and there is a need for agreed standards of research conduct. Nevertheless, several pharmacological agents have emerged as having potential. Acamprosate (a synthetic GABA analog) and naltrexone (an opioid anatagonist) have some evidence of efficacy in excessive drinking and alcohol dependence. However, precise mechanisms of effect and identification of drinkers most likely to benefit have still to be established. PMID- 19811022 TI - Behavioral treatment of insomnia. AB - Insomnia is a prevalent and potentially serious condition that compromises the functioning, health status and quality of lives of millions of individuals worldwide. Chronic insomnia is often perpetuated by psychological and behavioral factors, such as dysfunctional beliefs about sleep, heightened anxiety and sleep disruptive compensatory practices. Whereas sedative hypnotic medications are often prescribed for this condition, such treatment is symptom-focused and fails to address underlying factors sustaining the sleep problems. In contrast, behavioral insomnia therapies are specifically designed to address these perpetuating mechanisms. This article describes the main behavioral treatment approaches and their efficacy in treating insomnia. Current literature concerning treatment implementation factors (cost-effectiveness; treatment accessibility, delivery and dosage; patient response and adherence) is reviewed. PMID- 19811024 TI - Estrogen therapy in postmenopausal women with Parkinson's disease. PMID- 19811026 TI - Botulinum toxin type B (Myobloc, NeuroBloc): a new choice in cervical dystonia. AB - Botulinum toxin has dramatically improved the treatment of cervical dystonia. Prior to the use of botulinum toxin for many neurologic disorders, patients had few effective therapeutic options. Botulinum toxin type B (Myobloc, NeuroBloc) is a new antigenically distinct botulinum toxin with a unique structure and mechanism of action. Preclinical studies have demonstrated that im. injections of botulinum toxin type B effectively induce a dose-dependent paralysis. Controlled clinical trials have shown that it is safe and effective in alleviating symptoms associated with cervical dystonia. Given its efficacy and safety profile, the clinical use of type B toxin is anticipated to expand into other therapeutic areas. PMID- 19811027 TI - Galantamine: a new treatment for Alzheimer's disease. AB - Galantamine (Reminyl) is a tertiary alkaloid, originally isolated from the Caucasian snowdrop and several Amaryllidaceae plants. It has been used for over 40 years in anesthetics to reverse the effects of curarization. In vitro and in vivo studies have confirmed that galantamine is an orally-active, reversible, competitive inhibitor of brain cholinesterase, which is 50 times more selective for acetylcholinesterase, compared with butyrylcholinesterase. Galantamine has also been shown to allosterically potentiate sub-maximal nicotinic responses to acetylcholine. This direct effect on pre- and postsynaptic nicotinic receptors may represent a second mechanism of action for enhancing cholinergic function, but as yet it is unclear as to whether that has any clinical significance. Galantamine has been licensed for use in the symptomatic treatment of Alzheimer's disease patients, after proving its efficacy in a number of well-designed clinical trials. It has to be administered twice daily and has shown good tolerability, which typically for drugs of this class, is improved by slow titration. The most common side effects are: nausea, vomiting, diarrhea and anorexia. Galantamine appears to be an effective treatment for Alzheimer's disease comparable with the other acetylcholinesterase inhibitors in improving cognition and function with growing evidence for its effects on behavior and caregiver burden. PMID- 19811028 TI - Comparison of older and newer neuroleptics for the treatment of schizophrenia. AB - This article reviews the current antipsychotic treatments available for schizophrenia. After a review of its history - from the beginning of last century - emphasis is placed on the distinguishing features of the new antipsychotic therapies against typical neuroleptic agents. An attempt is made to relate the different mechanisms of antipsychotic events in these groups of drugs with their efficacy and safety profiles before going on to develop more extensively the prospects provided by new antipsychotic drugs in the areas of acute clinical efficacy and maintenance, safety and quality of life for the schizophrenic patient. PMID- 19811029 TI - Newer antipsychotics: comparative review of drug interactions. AB - The new atypical antipsychotics are subject to drug-drug interactions with other psychotropic agents or with medications used in the treatment of concomitant somatic illnesses - usually at the pharmacokinetic or pharmacodynamic level. While novel antipsychotics are unlikely to interfere with the elimination of other drugs, coadministration of inhibitors or inducers of the cytochrome P450 isoenzymes responsible for their metabolism may modify plasma antipsychotic concentrations, leading to potentially (clinically) significant effects. Newer antipsychotics have binding affinity at a variety of neurotransmitter receptors and might therefore be involved in pharmacodynamic interactions when given in combination with agents acting on the same systems. Differences in the interaction potential among the current novel antipsychotics may be predicted based on their pharmacokinetic and pharmacodynamic properties. Avoidance of unnecessary polytherapy, knowledge of the interaction profiles of individual agents and careful individualization of dosage based on close evaluation of clinical response and possibly plasma drug concentrations are essential to prevent and minimize potentially adverse drug interactions in patients receiving newer antipsychotics. PMID- 19811030 TI - Risk factors for schizophrenia. AB - Over the last decade, much progress has been made towards identifying risk factors for schizophrenia. It is now thought that many genes of small effect contribute towards risk of developing schizophrenia and some of these probably confer susceptibility to environmental factors. Large population-based studies, using record linkage, have investigated environmental risk factors and found that urbanization is an important factor. Other population-based studies have followed up birth cohorts and found evidence for abnormal neurodevelopment in those who subsequently developed schizophrenia. Meta-analyses have clarified the role of obstetric and perinatal complications especially hypoxia. Low IQ has emerged as a risk factor and suggests that cognitive factors are important in the etiology of psychosis. Social risk factors are now being investigated with some evidence that discrimination and isolation are significant. The role of recreational drug use is still unclear. Finally, evidence suggests that there is a continuum of psychosis both within the disorders that present clinically, but also in the general population. PMID- 19811031 TI - Teratogenesis with antiepileptic drugs: practical management of young women with epilepsy. AB - Children born to mothers who take antiepileptic drugs during pregnancy have a 2-4 fold increased risk for birth defects and also a risk of growth retardation and possibly impaired psychomotor development. Despite their teratogenic effects, antiepileptic drugs are used during pregnancy because uncontrolled epileptic seizures are considered more hazardous to the fetus than the drug therapy. The aim is to maintain seizure control with the appropriate drug given in monotherapy at the lowest effective dosage. These issues have implications for the management of young women with epilepsy in general, since major changes in drug therapy and other attempts to reduce the teratogenic risks should be accomplished before conception in order to be effective and safe. PMID- 19811032 TI - Antiepileptic drugs in the treatment of neuropathic pain. AB - Antiepileptic drugs were initially designed to treat epileptic seizures, but are increasingly used in the treatment of neuropathic pain. Efficacy of the newer antiepileptic drugs has been confirmed in placebo-controlled studies for trigeminal neuralgia, postherpetic neuralgia, painful polyneuropathy and central poststroke pain. While standard antiepileptic drugs act as sodium channel blockers, the newer antiepileptics display additional mechanisms of action, including a potentiation of GABAergic inhibition, specific binding on a subtype of calcium channels, a reduced release of glutamate and a blockade of AMPA/kainate receptors. These mechanisms could account for their antiallodynic and antihyperalgesic effects demonstrated in animal models of neuropathic pain and also suggested for gabapentin and lamotrigine in humans. Additional studies are required to compare efficacy of newer versus standard antiepileptic drugs and to better define their place in comparison with other analgesics in the treatment of neuropathic pain. PMID- 19811033 TI - Metabotropic glutamate receptors: novel targets for pain relief. AB - Metabotropic glutamate receptors (mGluRs) are a relatively new family of G protein coupled receptors that can be activated by the major excitatory neurotransmitter, L-glutamate. The eight known mGluR subtypes are classified into three groups based on their sequence homology, signal transduction mechanisms and receptor pharmacology. Extensive research has implicated mGluRs in neuroplasticity associated with normal brain functions, but also in various neurological and psychiatric disorders. Evidence is accumulating to suggest an important role of mGluRs in nociception and pain. With the availability in recent years of selective pharmacological tools, behavioral and electrophysiological studies have shown that mGluR subgroups and subtypes mediate and modulate nociceptive processing at different levels of the nervous system: periphery, spinal cord and brain. Thus, mGluRs may provide important novel therapeutic targets for the relief of pain associated with altered neurotransmission and neuronal excitability. PMID- 19811034 TI - Diabetic neuropathy: pathogenetic background, current and future therapies. AB - Diabetic neuropathy is the most common complication in diabetes. To date, there are few efficacious therapies for this complication. Several groups of drugs have undergone clinical testing, the outcome of which has not yielded convincing benefits. Diabetic neuropathy is a dynamic chronic degenerative disorder with multiple interactive pathogenetic mechanisms complicating the design of biologically-meaningful therapies. New insights into this disorder and the fact that it differs in the two types of diabetes has drawn attention to the etiological effects of insulin/C-peptide deficiency in diabetic neuropathy. Recent data regarding regulation of the aldose reductase gene has fueled new interest in aldose reductase inhibitors. A more downstream metabolic abnormality, oxidative stress, provoked by several mechanisms has become a therapeutic target of great interest. Therefore, despite previous therapeutic failures, there is now a renewed interest and reassessment of old therapies, as well as development of totally new strategies. PMID- 19811035 TI - Surgical treatment of Parkinson's disease: ablation, stimulation and transplantation. AB - Clinicians now have an extensive and effective pharmacopeia to treat the symptoms of Parkinson's disease - especially in the early and moderate stages of the disorder. Ultimately, a significant number of patients with the disorder find that, as their disease progresses, pharmacotherapy fails to control symptoms or produces unacceptable adverse effects as higher medication doses are used. For patients with Parkinson's disease whose symptoms are insufficiently controlled by medical therapy, surgical treatments can reduce symptoms, enhance functional capacity and sometimes reduce medication requirements. This article reviews the current state-of-the-art in surgical interventions for Parkinson's disease, with an emphasis on deep brain stimulation therapy, an evolving treatment that accomplishes symptom control in a reversible, adjustable and nondestructive manner. PMID- 19811036 TI - Inherited muscle and brain channelopathies. AB - In the past 5 years, advances in the complementary fields of neurogenetics and cellular electrophysiology have resulted in an explosion of knowledge about a group of disorders now known as the neurological channelopathies. These advances have resulted in more accurate DNA-based diagnosis and have increased our understanding of cellular pathophysiology. This is leading to more tailored therapies for patients with these disorders. PMID- 19811037 TI - Stem cells as therapeutics for neurodegenerative disorders? AB - Aging is associated with a progressive increase in the risk of several prominent neurodegenerative disorders, including Alzheimer's disease, Parkinson's disease, Huntington's disease, stroke and amyotrophic lateral sclerosis. In each of these disorders specific populations of neurons become dysfunctional, then die and are not replaced. The adult brain and spinal cord contain populations of so-called neural stem cells (self-renewing and multipotent) and neural precursor cells (specified to a certain fate, but still mitotic) that may provide a continuing source of new neurons and glial cells during successful aging and after injury to the nervous system. Recent studies have shown that stem cells from embryos and adults can be transplanted into the nervous system, differentiate into neurons and glia and restore lost function in experimental models of neurodegenerative diseases. Embryonic stem cells may be a particularly effective donor cell type for transplantation-based therapies. Efficacy of stem cell therapies remains to be established in clinical trials in humans. Another approach is to mobilize endogenous neural stem cells. Animals studies have shown that dietary and behavioral modifications can indeed stimulate neurogenesis. Molecular and cellular mechanisms that regulate the proliferation, differentiation and survival of neural stem cells and neural precursor cells are being elucidated and are revealing novel targets for the development of pharmaceuticals that promote neurogenesis. PMID- 19811040 TI - Challenging views of Alzheimer's disease. PMID- 19811038 TI - Will neurogenomics revolutionize neurotherapeutics? PMID- 19811041 TI - Donepezil in the treatment of patients with Alzheimer's disease. AB - Donepezil is the most widely used acetylcholinesterase inhibitor licensed for symptomatic treatment of mild-to-moderate Alzheimer's disease. Clinical placebo controlled trials and open-label extension studies have consistently shown that donepezil is well tolerated and gives rise to statistically significant improvements in cognition, global function and activities of daily living for at least 12 months and to less deterioration of function possibly for more than 4 years. Furthermore, donepezil may reduce neuropsychiatric symptoms and caregiver burden. Health economic studies suggest that treatment with donepezil may reduce resource utilization. Evidence-based international management guidelines recommend that treatment with a cholinesterase inhibitor should be considered in all patients with mild-to-moderate Alzheimer's disease and based on a proper diagnostic evaluation. Treatment cannot replace continuous advice to patients and caregivers. PMID- 19811042 TI - Almotriptan: a novel 5-HT1B/D agonist for the symptomatic treatment of migraine. AB - Currently available oral triptans are not ideal, at least for 20-30% of migraine patients, due to either response failure or adverse events. Almotriptan is a novel selective 5-HT1B/D receptor agonist exhibiting the highest bioavailability among triptans, both outside and within a migraine attack. The tolerability of the therapeutic oral dose of almotriptan, 12.5 mg, is similar to that of placebo, with a remarkably low incidence of chest symptoms. At this dose, efficacy parameters remain comparable to those of sumatriptan 100 mg, while the recurrence rate is in the lower range. This balanced profile makes almotriptan 12.5 mg a good choice for the symptomatic treatment of the typical migraine patient. PMID- 19811043 TI - IM olanzapine in the treatment of agitation and aggression. AB - Agitation or aggressive behaviors are frequently associated with an acute psychotic episode. Patients experiencing severe auditory hallucinations, paranoia or acute mania are particularly prone to agitation. In the elderly, agitation and aggression are often associated with delirium, dementia or various medical illnesses. Agitated patients urgently require rapid control of symptoms. Traditionally, treatment has included administration of iv. or im. benzodiazepines or neuroleptics. However, each of these is associated with adverse effects that may be serious in nature. An injectable formulation of a novel antipsychotic may effectively treat agitation and aggression while placing the patient at a lower risk for more serious side effects frequently associated with parenteral neuroleptics. PMID- 19811044 TI - Lamotrigine. AB - Lamotrigine is a newly available antiepileptic drug with a broad spectrum of efficacy and a tolerability profile that is favorable for use in many patients. Problems with rash are related to rate of dose ascension and drug interactions with valproate. Efficacy in treatment of mood disorder and some pain syndromes will broaden the use of this unique compound. PMID- 19811045 TI - Pramipexole: a nonergot dopamine agonist as drug therapy in Parkinson's disease. AB - Pramipexole is a dopamine agonist that was recently introduced for the treatment of both early and advanced Parkinson's disease. Pramipexole is a synthetic aminobenzothiazole compound with a high affinity to D(3) receptors. It directly stimulates dopamine receptors without requiring metabolic conversion. Pramipexole is rapidly absorbed with bioavailability greater than 90% and is actively secreted by the renal tubules. The drug's plasma half-life is 8-12 h. Shown to be safe and effective as add-on therapy, pramipexole can also be used as initial dopaminergic therapy in Parkinson's disease, delaying the introduction of levodopa thereby postponing levodopa's side-effects. PMID- 19811046 TI - Chronic inflammation in Alzheimer's disease offers therapeutic opportunities. AB - Postmortem studies have revealed a state of chronic inflammation in affected regions of the brain in Alzheimer's disease (AD). Chronic inflammation can be damaging to host cells and the brain may be particularly vulnerable as neurons are not replaced. Evidence suggests that the inflammation is killing neurons in AD brain, so anti-inflammatory agents might slow the process of the disease. More than 20 epidemiological studies have shown that persons taking nonsteroidal anti inflammatory drugs (NSAIDs) have a greatly reduced incidence of AD. In one clinical trial, indomethacin appeared to halt the progression of memory loss in AD patients. NSAIDs inhibit synthesis of prostaglandins, which are fringe players in the inflammatory process. Agents that would block the more important actors, such as the complement system, activated microglia and inflammatory cytokines, might have important therapeutic benefits in AD as well as in other conditions, such as heart disease and stroke, where inflammation also plays a deleterious role. PMID- 19811047 TI - Use of acetylcholinesterase inhibitors in Alzheimer's disease. AB - Alzheimer's disease is a growing problem in an aging Western world, estimated to have cost the US economy USD 1.75 trillion. Until recently, the management of Alzheimer's disease largely comprised support for the family, nursing care and the use of unlicensed medication to control behavioral disturbances. The three new acetylcholinesterase inhibitors licensed to treat Alzheimer's disease (donepezil, rivastigmine and galantamine) have provided clinicians with a major impetus to their desire to diagnose and treat this lethal disease. Their effects on cognition are proven. More recent work on the effects of acetylcholinesterase inhibitors on behavioral symptoms, activities of daily living and caregiver burden have also been encouraging. Emerging work indicates their likely efficacy in other dementias (e.g., vascular dementia, dementia with Lewy bodies). This review summarizes the evidence concerning the impact of acetylcholinesterase inhibitors in dementia both currently and over the next 5 years. PMID- 19811048 TI - Treatment of behavioral/psychological symptoms in Alzheimer's disease. AB - This review examines the evidence for the use of drugs as part of the treatment of depressive/psychotic symptoms and behavioral disturbances in persons with Alzheimer's disease. Although conventional antidepressants appear to relieve depressive symptoms in Alzheimer's disease patients, placebo seems equally effective. The use of most medications used to control psychotic symptoms and disturbed/disturbing behaviors in Alzheimer's disease is off-label and not strongly supported by randomized, placebo-controlled trials, especially for so called agitation. For these reasons, trials of psychotropic drugs in Alzheimer's disease need a placebo condition in addition to active comparators and in the case of nondrug treatments, a sham treatment condition. Cholinesterase inhibiting drugs may reduce behavioral symptoms in addition to slowing cognitive decline. Selective (for brain region) GABA agonists seem to offer the greatest theoretical promise for reducing agitated behaviors not driven by depression or psychosis. PMID- 19811049 TI - Contemporary and emergent pharmacological therapies for chronic pain: nonopioid analgesia. AB - The American Pain Society estimates that 50 million Americans are partially or totally disabled by pain. This striking statistic is certain to increase as our population continues to age. In order to combat this growing problem, healthcare professionals must arm themselves with information. By developing the appropriate pain assessment skills and by staying abreast of the rapidly-changing therapies used in pain management, clinicians can dramatically impact the quality of life of those living with pain. PMID- 19811050 TI - Cannabinoids and pain relief. AB - Understanding of the structure and function of the endocannabinoid system is rapidly evolving. Physiological and pharmacological manipulations based on cannabinoid receptors, ligands and endocannabinoids have explained some medicinal attributes of cannabinoids as used across the world for thousands of years. Plant derived and synthetic cannabinoids are available for therapeutic use. Small clinical trials have demonstrated analgesic potential in acute and chronic pain. Regulatory and pharmacological limitations of these agents have hindered pain research in humans. Selective agonists, antagonists and metabolic targets to enhance endogenous cannabinoid activity are in development. Government reports in Europe and North America have encouraged research into the use of cannabinoids for pain relief and endorsed the clinical trials in acute and chronic pain. The results of large clinical trials into cannabinoid use for acute pain is expected to be the catalyst for wider studies and possible changes in legislation. Long term effects of psychoactive cannabinoids require close monitoring and international cooperation to define their role, if any, in CNS disorders. PMID- 19811051 TI - Complex regional pain syndrome: an actual survey. AB - Complex regional pain syndrome - in the past called sympathetic reflex dystrophy is, in its pathophysiology, still not fully understood. However, research in the last few years has led to a better understanding of the illness and the beginning of a pathophysiologically- orientated therapy. The core hypothesis is based on neuropeptide release, neurogenic inflammation and its sympathetic dependence. Therapy should be based on current pathophysiological concepts regarding CRPS and neuropathic pain and could thereby lead to a good outcome of the illness. PMID- 19811052 TI - Neuroprotective therapy for Huntington's disease: new prospects and challenges. AB - Extraordinary advances in understanding of the molecular bases of neurodegeneration have occurred since the Huntington's disease genetic mutation was discovered. Many relevant routes to neuronal demise in Huntington's disease have been identified including: glutamatergic stress, metabolic insufficiency, oxidative stress, proapoptotic signaling, inflammatory signaling, altered proteolysis, protein aggregation, transcriptional dysregulation, abnormal protein folding and neurotrophin insufficiency. Each represents specific therapeutic opportunities, which are being tested in high-throughput screens as well as in genetic models of Huntington's disease, transgenic mouse models and human clinical trials. Challenges include the uncertain power of these preclinical studies to predict therapeutic efficacy in humans, prioritizing the many approaches for human clinical trials and learning how to perform neuroprotective trials in presymptomatic individuals while protecting them from unwanted genetic information. PMID- 19811053 TI - Management of Guillain-Barre syndrome. AB - Guillain-Barre syndrome is now the most common cause of acute flaccid paralysis. Although modern intensive care facilities have significantly reduced the mortality associated with this monophasic immune mediated neuropathy, up to 5% of patients do not survive and 10% are left unable to walk unaided. Supportive medical care remains the most important aspect of management but both plasmapheresis and iv. immunoglobulin have been shown to hasten recovery when given in the early stages of the illness. This article summarizes the current management of Guillain-Barre syndrome, including both supportive measures and immune modulating therapy. PMID- 19811055 TI - Aeromonas sepsis as a marker of devitalized tissue in traumatic injuries. PMID- 19811056 TI - Chronic surgical site infection due to suture-associated polymicrobial biofilm. AB - BACKGROUND: Surgical site infection (SSI) is a common surgical complication; culture-negative SSI presents a particular problem in management. METHODS: Examination of explanted foreign bodies (sutures) using confocal laser scanning microscopy (CLSM) and fluorescent in situ hybridization (FISH) after surgical exploration of a chronic culture-negative SSI. RESULTS: Confocal microscopy (CM) demonstrated bacilli and cocci attached to the surface of the explanted sutures in a mixed biofilm. Fluorescent in situ hybridization confirmed that Staphylococci were components of the mixed biofilm. Removal of the foreign bodies (sutures) resolved the chronic infection. CONCLUSION: Chronic SSI can arise from underlying bacterial biofilms, which can invest implanted foreign bodies and associated soft tissue surfaces. PMID- 19811057 TI - Use of a modified syringe barrel to ensure control of the Amplatz sheath during percutaneous nephrolithotripsy in obese patients. AB - Percutaneous nephrolithotripsy (PNL) has been shown to be safe and effective in obese patients. One technical problem specifically encountered in these patients is migration of the Amplatz sheath beneath the skin or muscle fascia. We describe a simple technique, making use of a modified 10-cc syringe barrel, to facilitate retrieval of a migrated access sheath in obese patients undergoing percutaneous nephrolithotripsy. This can also be adopted to prevent loss of the working sheath in the first place, as well as to provide some extra length to access the collecting system avoiding the need to convert to longer instrumentation. This technique is cheap, safe, and effective. It avoids the need to extend the skin incision, resulting in improved cosmesis and reduced postoperative pain. PMID- 19811058 TI - Experience and learning curve of retroperitoneal laparoscopic ureterolithotomy for upper ureteral calculi. AB - PURPOSE: To summarize our experience and evaluate the learning curve of retroperitoneal laparoscopic ureterolithotomy of the upper ureter. PATIENTS AND METHODS: Between May 2004 and May 2007, 40 patients underwent retroperitoneal laparoscopic ureterolithotomy of the upper ureter. We divided the first and last 20 patients into group I and group II. There was no statistical difference in stone size between groups. Operative time and complications were measured as a basis for the assessment of the learning curve. RESULTS: In group I, the complication rate was 15% (3/20), including two patients whose procedure was converted to open surgery because of intraoperative bleeding, and one patient who experienced urine leakage because of a displaced Double-J ureteral stent. In group II, no postoperative complications occurred, while the mean operative time was significantly shorter compared with the earlier operations (65 vs 120 min). CONCLUSION: Retroperitoneal laparoscopic ureterolithotomy is safe and effective for large or impacted stones of the upper ureter. It is associated with a short learning curve in the setting of an active laparoscopic practice for selected patients. PMID- 19811059 TI - Management of benign ureteral strictures in the endoscopic era. AB - BACKGROUND AND PURPOSE: During the past decade, endoscopic management has emerged as the first-line treatment of benign ureteral strictures. We reviewed our experience with the management of benign ureteral strictures to determine the success rate of endoscopic surgery in a contemporary series and assessed the viability of surgical reimplantation in the modern era. PATIENTS AND METHODS: We identified 75 patients with a diagnosis of ureteral stricture between 2000 and 2005 via electronic medical records search and excluded those with completely obliterated, external compressive, malignant, or ureteroenteric strictures, ureteropelvic junction obstruction, and those with follow-up less than 2 months. RESULTS: Thirty-four patients who were treated endoscopically (balloon dilation and/or holmium laser endoureterotomy) were identified. Mean stricture length in each patient was 1.6 +/- 1 cm (range 0.5-4 cm), and the mean number of procedures per patient was 1.7 +/- 0.8. Endoscopic success was achieved in 29 (85%), while 5 (15%) patients experienced endoscopic management failure and ultimately needed ureteral reimplantation. When comparing the endoscopically treated and reimplant groups, there was no significant difference in mean stricture length (1.38 +/- 1.13 vs 2 +/- 1.1 cm, P = 0.14), yet mean number of procedures performed (1.41 +/ 0.85 vs 3.6 +/- 1.5; P = 0.002) reached statistical significance. There were no clinical or radiographic signs of obstruction in 100% of patients who received endoscopic therapy only and 100% of patients who needed open surgical management at a mean follow-up of 25.2 +/- 19.3 and 7.7 +/- 3.2 months, respectively. CONCLUSIONS: Endoscopic surgery is clearly a successful primary treatment modality in the management of benign ureteral strictures with minimal morbidity. In the modern era of endoscopic surgery, however, ureteral reimplantation remains a viable option in treating the small subset of patients with benign ureteral strictures for whom endoscopic management fails. PMID- 19811060 TI - Factors affecting the fluoroscopic screening time during percutaneous nephrolithotomy. AB - PURPOSE: Percutaneous nephrolithotomy (PNL) is established in urology practice with routine use of fluoroscopic guidance. Herein, we retrospectively evaluated the potential factors affecting the fluoroscopic screening time (FST) during PNL. MATERIALS AND METHODS: A total of 282 patients with documented FST in detail during PNL performed by one surgeon were enrolled in this study. In all cases, PNL was performed on patients in prone position under C-armed fluoroscopy unit. The correlation between the FST and patient and also stone factors, including body mass index (BMI), previous open renal surgery, characteristics and size of the stone, and presence of hydronephrosis, as well as procedural factors such as percutaneous access number and location and operation time were analyzed. RESULTS: The mean age of the patients was 43 +/- 14.7 (range: 18-67) years. The mean operation time was 78.9 +/- 22.0 (range: 45-150) minutes, and the mean FST was 10.19 +/- 6.3 (range: 3-50) minutes. The mean stone size was 8.46 +/- 5.11 cm(2) (range: 2-30). The FST was significantly prolonged in patients with increased stone burden (p = 0.001) and necessitating multiple accesses procedures (p = 0.007). However, BMI of the patient, stone configuration, degree of hydronephrosis, history of open renal surgery, and access location did not have an impact on FST (p > 0.05). CONCLUSION: In patients with large stones necessitating multiple accesses, FST is significantly prolonged; therefore, they have to be managed by experienced surgeons to diminish the radiation exposure. However, BMI, stone configuration, presence of hydronephrosis, history of open renal surgery, and access location did not have effect on FST in our study. PMID- 19811061 TI - Subcutaneous fixation of gastrostomy tube is superior to temporary fixation. AB - PURPOSE: The placement of gastrostomy tubes (GTs) in infants and children to provide enteral access over the past decade has shifted toward a minimally invasive, safer direction with the development of various new techniques. We have developed a modified technique, utilizing subcutaneous (S.C.) tunneling stay sutures to prevent complications, such as GT dislodgment and wound infection. The aim of this study was to identify and describe complications of the modified procedure and compare it with the standard laparoscopic GT placement. MATERIALS AND METHODS: A retrospective 4-year review of 153 patients who underwent laparoscopic GT placement was conducted. Eighty-nine patients underwent the modified S.C. tunneled technique, and 64 patients had standard U-stitch with temporary fixation. RESULTS: The overall complication rate was significantly higher with the temporary fixation group (20%) versus the modified technique group (2.5%). Six patients from the temporary group and 2 patients from the modified group developed cellulitis. Seven patients underwent reoperation secondary to GT dislodgment in the temporary group. No patients were identified with tube-related pressure necrosis or procedure-related deaths. CONCLUSION: S.C. placement of stay sutures in the modified technique is associated with a lower complication rate than temporary fixation. The modified technique should be the preferred approach when performing laparoscopic GT insertion. PMID- 19811062 TI - Pediatric laparoscopic-assisted gastric transposition: early experience and outcomes. AB - astric transposition in children is a well-established treatment for long-gap esophageal atresia and long caustic strictures when preservation of the native esophagus is impossible, but does involve laparotomy and, often, thoracotomy incisions. The minimally invasive approach to this operation has been attempted in 7 patients to date in the UK, all at Great Ormond Street Hospital (London, UK). Four patients were male, 3 were female, with a mean age of 3 years and 7 months (range, 5 months to 13 years). Indications were long-gap esophageal atresia where primary anastomosis was impossible (5 patients) and caustic ingestion with long esophageal strictures (2 patients). The stomach was mobilized laparoscopically by using 5-mm instruments and a pneumoperitoneum of 10 mm Hg. Following pyloromyotomy or pyloroplasty, a tunnel in the posterior mediastinum was developed. The fundus of the stomach was drawn to the neck and sutured to the cervical esophagus. The patients were ventilated on our intensive care unit and then returned to the surgical ward before discharge. Gastric transposition was completed laparoscopically in 6 of 7 patients. The mean duration of surgery was 327 minutes (range, 240-455). All patients survived. Minor complications were pneumonia (2), pleural effusion (1), and hiatus hernia (2). The mean hospital stay was 16 days (range, 8-36). Our preliminary experience would suggest that laparoscopic gastric transposition is a safe alternative to open surgery with satisfactory results. PMID- 19811063 TI - Video-assisted thoracoscopy in compromised pediatric patients. AB - The use of video-assisted techniques (VATs) in the paediatric field has become increasingly more frequent, based on reports of prompter recovery following VATs in respect to standard techniques. Specific advantages have been documented, in particular for pediatric patients undergoing chemioradiotherapic treatment. We retrospectively reviewed data of severely compromised patients who underwent VAT for lung wedge resections and biopsies carried out in our center over a 7-year period. As far as the area of therapeutic tumor resection is concerned, the present data are consistent with the view that thoracoscopy is both an effective and safe tool in diagnostic procedures. PMID- 19811064 TI - Laparoscopic cholecystectomy in the obese: results with the traditional and fundus-first technique. AB - INTRODUCTION: The aim of this study was to assess retrospectively the results of laparoscopic cholecystectomy (LC) performed in obese patients at our institution with the traditional technique and with the fundus-first (FF) technique. PATIENTS AND METHODS: We performed a retrospective analysis of 194 obese patients that underwent LC between 1994 and December 2007 at our institution. Surgical techniques were compared with respect to operative times, conversion to open cholecystectomy, postoperative complications, mortality, and length of postoperative stay. RESULTS: In the reviewed period, LC was performed in 113 (58.2%) patients with obesity type I (OTI), 55 (28.3%) patients with obesity type II (OTII), and 26 (13.5%) patients with obesity type III (OTIII). None of the differences among obese groups treated with the two techniques were statistically significant, with the exception of the lower operative times in the OTIII patients treated with the FFLC. The median operating time in the OTIII group was, respectively, 90 minutes for traditional LC and 65 (range, 45-130) for FFLC (P < 0.05). DISCUSSION AND CONCLUSIONS: This study achieved to conclude that LC in the obese is a safe, feasible, and efficient operation, but remains a demanding procedure even in experienced hands. FFLC can support the traditional LC in the treatment of obese patients, yielding a complication rate comparable with the traditional technique. In our study, it significantly reduced the operative time in OTIII patients, simplifying all the intra-abdominal maneuvers and the gallbladder dissection. PMID- 19811066 TI - Role of proteomics in nutrigenomics and nutrigenetics. PMID- 19811065 TI - Extraluminal laparoscopic wedge-resection of submucosal tumors on the posterior wall of the gastric fundus close to the esophagocardiac junction. AB - PURPOSE: Laparoscopic resection of submucosal tumors in the gastric fundus, especially in the posterior wall near the esophagocardiac junction (ECJ), is difficult and time consuming and is and likely to cause esophageal stenosis and splenic injury. In this article, we report an extraluminal laparoscopic wedge resection (ELWR) that minimizes these problems. METHODS: Thirty-seven patients with submucosal tumors in the posterior wall of the gastric fundus received ELWR. The operation consisted of four steps: 1) localization of the tumor, 2) dissection of the omentum, 3) mobilization of the gastric fundus/upper pole of the spleen and exposure of the ECJ, and 4) resection of the gastric fundus with a linear endoscopic gastrointestinal anastomosis stapler. RESULTS: None of the cases needed conversion to open surgery. Mean postoperative hospital stay was 5.5 +/- 1.0 days. The distance between the tumor and the incision margin ranged from 0.7 to 2.5 cm toward the ECJ. Pathologic examination revealed 7 cases of leiomyomas, 29 cases of stromal tumors (4 were low-grade malignant tumors), and 1 case of neurofibroma. There was no recurrence, metastasis, esophageal stenosis, or any other severe adverse event during the follow-up period (52 +/- 3.1 months). CONCLUSIONS: ELWR is a safe, effective treatment for submucosal tumors in the posterior wall of the gastric fundus. PMID- 19811067 TI - Mass spectrometry-based clinical proteomics profiling: current status and future directions. PMID- 19811068 TI - Vascular proteomics, a translational approach: from traditional to novel proteomic techniques. PMID- 19811070 TI - Framework for a systems approach to proteomic biomarker profiling in polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is the most common endocrine disorder in females of reproductive age, and its prevalence ranges between 6 and 8%. Associated problems include infertility, menstrual disorders, hirsutism and obesity. In addition, individuals with PCOS may be at increased risk of diabetes, endometrial cancer and, possibly, cardiovascular disease and breast cancer in later life. Biomarkers identified from proteomic analyses may help to improve the clinical management of PCOS, provided that new proteomic data can be integrated with existing knowledge and/or pathways implicated in disease etiology. In this study, a database of identity, descriptions and functions/pathways has been developed from 148 published proteomic biomarkers in PCOS. From analysis of the database, a variety of pathways possibly implicated in PCOS were determined, including those related to fibrinolysis, thrombosis, the antioxidant pathway and the immune system. This database, if developed further, will provide a framework for a systems approach to profiling biomarkers in the future. PMID- 19811071 TI - Relieving the first bottleneck in the drug discovery pipeline: using array technologies to rationalize membrane protein production. AB - The slow down in the drug discovery pipeline is, in part, owing to a lack of structural and functional information available for new drug targets. Membrane proteins, the targets of well over 50% of marketed pharmaceuticals, present a particular challenge. As they are not naturally abundant, they must be produced recombinantly for the structural biology that is a prerequisite to structure based drug design. Unfortunately, however, obtaining high yields of functional, recombinant membrane proteins remains a major bottleneck in contemporary bioscience. While repeated rounds of trial-and-error optimization have not (and cannot) reveal mechanistic details of the biology of recombinant protein production, examination of the host response has provided new insights. To this end, we published an early transcriptome analysis that identified genes implicated in high-yielding yeast cell factories, which has enabled the engineering of improved production strains. These advances offer hope that the bottleneck of membrane protein production can be relieved rationally. PMID- 19811073 TI - Cell-type-specific proteome and interactome: using HIV-1 Tat as a test case. AB - HIV-1 is a small retrovirus that wreaks havoc on the human immune system. It is a puzzle to the scientific community how a virus that encodes only nine proteins can take complete control of its host and redirect the cell to complete replication or maintain latency when necessary. One way to explain the control elicited by HIV-1 is through numerous protein partners that exist between viral and host proteins, allowing HIV-1 to be intimately involved in virtually every aspect of cellular biology. In addition, we postulate that the complexity exerted by HIV-1 can not merely be explained by the large number of protein-protein interactions documented in the literature but, rather, cell-type-specific interactions and post-translational modifications of viral proteins must be taken into account. We use HIV-1 Tat and its influence on viral transcription as an example of cell-type-specific complexity. The influence of post-translational modifications (acetylation and methylation), as well as subcellular localization on Tat binding partners, is also discussed. PMID- 19811072 TI - Urinary proteomic profiling for diagnostic bladder cancer biomarkers. AB - The ability to detect and monitor bladder cancer in noninvasively obtained urine samples is a major goal. While a number of protein biomarkers have been identified and commercially developed, none have greatly improved the accuracy of sample evaluation over invasive cystoscopy. The ongoing development of high throughput proteomic profiling technologies will facilitate the identification of molecular signatures that are associated with bladder disease. The appropriate use of these approaches has the potential to provide efficient biomarkers for the early detection and monitoring of recurrent bladder cancer. Identification of disease-associated proteins will also advance our knowledge of tumor biology, which, in turn, will enable development of targeted therapeutics aimed at reducing morbidity from bladder cancer. In this article, we focus on the accumulating proteomic signatures of urine in health and disease, and discuss expected future developments in this field of research. PMID- 19811074 TI - In silico methods for predicting T-cell epitopes: Dr Jekyll or Mr Hyde? AB - In silico tools offer an attractive alternative strategy to the cumbersome experimental approaches to identify T-cell epitopes. These computational tools have metamorphosed over the years into complex algorithms that attempt to efficiently predict the binding of a plethora of peptides to HLA alleles. In recent years, the scientific community has embraced these techniques to reduce the burden of wet-laboratory experimentation. Although there are some splendid examples of the utility of these methods, there are also evidences where they fall short and remain inconsistent. Hence, are these computational tools 'Dr Jekyll' or 'Mr Hyde' to the researcher, who wishes to utilize them intrepidly? This article reviews the progress and pitfalls of the in silico tools that identify T-cell epitopes. PMID- 19811075 TI - Role of proteomic technologies in understanding risk of arterial thrombosis. AB - Arterial thrombosis is a pivotal event in the development of cardiovascular diseases. Plasma and cellular proteins have the potential to influence thrombus morphology and function. This review summarizes the latest studies to use proteomic technologies to characterize the cellular and plasma components involved in arterial thrombosis, with a view to understanding the pathogenesis and treatment of acute cardiovascular diseases. Proteomic approaches have been extensively used to profile the proteome of endothelial cells, leukocytes, vascular smooth muscle cells, platelets and plasma in the search for risk factors for cardiovascular disease; however, further work is required to validate the direct contribution of these proteins to arterial thrombosis. PMID- 19811076 TI - Current advances in tumor proteomics and candidate biomarkers for hepatic cancer. AB - Tumor proteomics apply proteomics techniques to tumor biological research, mainly by screening candidate biomarkers for early tumor diagnosis, prognosis and treatment. Hepatocellular carcinoma (HCC) is a type of malignant tumor with one of the highest death rates in the world. With the advent of the post-genomic age, tumor biological research developing the technology of proteomics has become a major focus of researchers. The discovery of novel candidate biomarkers is one of crucial problems for the early diagnosis of HCC. In general, there are three distinct types of candidate biomarkers for HCC based on different areas: biochemical biomarkers, antigenic biomarkers and epigenetic biomarkers. This review mainly discusses current advances in the problems and prospects of candidate biomarker for the early diagnosis of HCC, discovered by technologies of tumor proteomics. PMID- 19811077 TI - Disulfide-related proteomic studies on food allergens. AB - An increasing proportion of the population is affected by food allergy. Some food allergens ('incomplete' allergens) elicit clinical symptoms only owing to their homology with other allergens, while others ('complete') can both sensitize and elicit clinical symptoms. To challenge food allergy, it should be effective to invest more energy in tackling the complete ones. Post-translational modification proteomics allows a comprehensive investigation of complete allergens. It provides clues to elucidate mechanisms of the structures that contribute to allergenicity, which thus, in turn, to help alleviate food allergens. This review summarizes the recent proteomic challenges to identify and alleviate food allergens, especially through disulfide-related studies. PMID- 19811078 TI - Emerging affinity-based techniques in proteomics. AB - Proteomes of interest, such as the human proteome, have such complexity that no single technique is adequate for the complete analysis of the constituents. Depending on the goal (e.g., identification of a novel protein vs measurement of the level of a known protein), the tools required can vary significantly. While existing methods provide valuable information, their limitations drive the development of complementary, innovative methods to achieve greater breadth of coverage, dynamic range or specificity of analysis. We will discuss affinity based methods and their applications, focusing on their unique advantages. In addition, we will describe emerging methods with potential value to proteomics, as well as the challenges that remain for proteomic studies. PMID- 19811080 TI - Effectiveness of 980-mm diode and 1064-nm extra-long-pulse neodymium-doped yttrium aluminum garnet lasers in implant disinfection. AB - OBJECTIVE: To evaluate the potential of 980-nm gallium aluminum arsenide (GaAlAs) and 1064-nm neodymium-doped yttrium aluminum garnet (Nd:YAG) lasers to reduce bacteria after irradiation of implant surfaces contaminated with Enterococcus faecalis and Porphyromonas gingivalis and on irradiated implant surface morphology. BACKGROUND: Despite the frequency of implant success, some implant loss is related to peri-implantitis because of difficulty in eliminating the biofilm. METHODS: Implants (3.75 x 13 mm) with machined surfaces, surfaces sand blasted with titanium oxide (TiO(2)), and sand-blasted and acid-etched surfaces were exposed to P. gingivalis and E. faecalis cultures and irradiated with 980-nm GaAlAs or 1064-nm Nd:YAG lasers. After laser treatments, the number of remaining colony-forming units and implant surface morphology were analyzed using scanning electron microscopy (SEM). RESULTS: The Nd:YAG laser was able to promote a total contamination reduction on all implants irradiated. The results with the GaAlAs laser showed 100% bacteria reduction on the implants irradiated with 3 W. Irradiation with 2.5 W and 3 W achieved 100% of bacteria reduction on P. gingivalis-contaminated implants. Decontamination was not complete for the sand blasted TiO(2) (78.6%) and acid-etched surfaces (49.4%) contaminated with E. faecalis and irradiated with 2.5 W. SEM showed no implant surface changes. CONCLUSION: The wavelengths used in this research provided bacteria reduction without damaging implant surfaces. New clinical research should be encouraged for the use of this technology in the treatment of peri-implantitis. PMID- 19811081 TI - A comparative study of the effects of ab externo superpulse carbon dioxide laser assisted trabeculectomy with conventional trabeculectomy in rabbits. AB - OBJECTIVE: The aim of this investigation was to evaluate the efficacy of reduction in intraocular pressure (IOP) and complications in ab externo superpulse carbon dioxide laser-assisted trabeculectomy. BACKGROUND: Reduction of IOP remains the immediate goal of glaucoma treatment. Although trabeculectomy has been considered the gold standard for the surgical treatment of glaucoma, many complications are associated with this procedure. Laser surgery has been studied as an alternative way to control IOP in glaucoma. MATERIALS AND METHODS: Sixty adult albino rabbits were enrolled in this study. The animals underwent one unilateral surgery of either trabeculectomy or ab externo superpulse carbon dioxide laser-assisted trabeculectomy and were divided into two subgroups, control or experimental. Follow-up examinations included ocular anterior segments, filtration blebs and IOP. Five eyes in each group were analyzed histologically one month postoperatively. RESULTS: Anterior chamber hyphema in the laser group was milder than that in the surgery group, and disappeared on the third day. In the surgery group, it disappeared by the end of the first month. Filtration blebs in the laser group formed earlier than in the surgery group. Comparing the laser group and the surgery group, IOP values in the laser group were more stable. Histological examination showed that the sclera tract was blocked by scar tissue, inflammation had infiltrated, and the anterior chamber angle had adhered in the surgery group. CONCLUSIONS: Ab externo superpulse carbon dioxide laser-assisted trabeculectomy was suggested as a feasible technique to control IOP with the advantages of more steady IOP reduction and fewer complications. PMID- 19811082 TI - The effect of laser irradiation on proliferation of human breast carcinoma, melanoma, and immortalized mammary epithelial cells. AB - OBJECTIVE: This study compared the effects of different doses (J/cm(2)) of laser phototherapy at wavelengths of either 780, 830, or 904 nm on human breast carcinoma, melanoma, and immortalized human mammary epithelial cell lines in vitro. In addition, we examined whether laser irradiation would malignantly transform the murine fibroblast NIH3T3 cell line. BACKGROUND: Laser phototherapy is used in the clinical treatment of breast cancer-related lymphoedema, despite limited safety information. This study contributes to systematically developing guidelines for the safe use of laser in breast cancer-related lymphoedema. METHODS: Human breast adenocarcinoma (MCF-7), human breast ductal carcinoma with melanomic genotypic traits (MDA-MB-435S), and immortalized human mammary epithelial (SVCT and Bre80hTERT) cell lines were irradiated with a single exposure of laser. MCF-7 cells were further irradiated with two and three exposures of each laser wavelength. Cell proliferation was assessed 24 h after irradiation. RESULTS: Although certain doses of laser increased MCF-7 cell proliferation, multiple exposures had either no effect or showed negative dose response relationships. No sign of malignant transformation of cells by laser phototherapy was detected under the conditions applied here. CONCLUSION: Before a definitive conclusion can be made regarding the safety of laser for breast cancer related lymphoedema, further in vivo research is required. PMID- 19811083 TI - Venous lake of the lips treated using photocoagulation with high-intensity diode laser. AB - OBJECTIVE: To evaluate the effectiveness of photocoagulation with high-intensity diode laser in the treatment of venous lake (VL) lesions. BACKGROUND DATA: VL is a common vascular lesion characterized by elevated, usually dome-shaped papules, ranging in color from dark blue to dark purple, seen more frequently in elderly patients. They often occur as single lesions on the ears, face, lips, or neck. Once formed, lesions persist throughout life. Although these lesions are usually asymptomatic, they can bleed if injured. METHODS: Seventeen patients (7 men and 10 women) with VL on the lip were treated using a noncontact diode laser (wavelength 808 nm, power output 2-3 W in continuous wave). RESULTS: After only one irradiation exposure, all lesions were successfully treated. Healing was completed in approximately 2 to 3 weeks, and none of the patients experienced complications. Postoperative discomfort and scarring were not present or were minimal. CONCLUSION: Photocoagulation with high-intensity diode laser is an effective, bloodless procedure for the treatment of VL. PMID- 19811084 TI - A focus on clinical immunology. PMID- 19811085 TI - Fine specificity of autoantibodies induced by mouse hepatitis virus A59. AB - We have shown that mice infected with mouse hepatitis virus A59 (MHV-A59) develop autoantibodies (autoAb) to liver and kidney fumarylacetoacetate hydrolase (FAH). The autoAb recognized conformational as well as linear antigenic determinants in the enzyme, and the autoimmune response was not entirely restricted to molecular mimicry and/or epitope spreading. Since the N- and C-terminal portions of the enzyme were the most reactive with autoAb, the fine specificity of these Ab was investigated. Immobilized 15-mer linear peptides (overlapping by 14 amino acids) spanning the N-terminal FAH sequence 1-49 were recognized by Ab from MHV-infected mice. The pattern of reactivity indicated the existence of two major epitope cores (i.e., sequences 9-23 and 30-44), and sequence comparison permitted the identification of two minimal epitopes, DSDFPIQ (amino acids 9-15) and IGDQILD (amino acids 36-42). Mutational analysis of sequences 9-23 and 30-44 indicated that residues 9-12 (DSDF) from the first major N-terminal epitope, and residue 36 (I) from the second, were the key amino acids energetically important for Ab contact. Interestingly, those residues were inside the two minimal epitopes previously predicted. The C-terminal portion of the enzyme (sequence 390-419) presented only one major epitope, located between residues 390 and 409. In this case, the minimal epitope had nine amino acids, CQGDGYRVG, corresponding to the FAH sequence 396-404 that outlines a loop specific for the enzyme. Data indicated that neither the FAH minimal epitopes nor the key residues important for binding to Ab from MHV-infected mice have their counterparts in the viral proteins. However, location of the energetically important residues in the tertiary structure of the enzyme originates a virtual conformational epitope. Such hypothetical B-cell epitopes could be present in any viral protein, originating a cross-reaction leading to the autoimmune response induced by MHV. PMID- 19811086 TI - Downregulation of HLA class II molecules by G1896A pre-core mutation in chronic hepatitis B virus infection. AB - Over the past decade, increasing attention has been focused on the contribution of naturally occurring mutations in the hepatitis B virus (HBV) genome to the clinical course of the chronic infection. The aim of this study was to investigate the effect of the HBV pre-core mutation G1896A on the expression of HLA class II molecules and the core protein of hepatitis B in liver biopsies of chronic hepatitis B (CHB) infection. In 30 HBeAg-negative CHB patients the pre core region of the HBV genome was amplified and sequenced to determine the presence of the mutation G1896A. Liver biopsies were scored based on the Histology Activity Index (HAI) system and immunohistochemistry (IHC) was performed to study the expression of HLA class II molecules on the antigen presenting cells and the core protein in hepatocytes. We found that 19 of the 30 patients (63%) harbored the G1896A mutation. Compared to the patients without this mutation, those with G1896A had lower HAI scores (5.0 +/- 2.8 versus 7.9 +/- 4, p = 0.03). The study of the expression of HLA-II molecules in our patients revealed that subjects with the G1896A mutation had lower expression of HLA-II compared to wild-type infected subjects (1.87 +/- 0.6 versus 3.27 +/- 1.5, p < 0.01). Core protein expression was present in four patients (13.3%) who had higher HBV DNA levels than patients without core protein expression (3.81 +/- 0.78 versus 2.02 +/- 0.16, p < 0.001). These results suggest that the G1896A pre core mutation may directly interfere with antigen presentation, most likely by decreasing the availability of HLA class II molecules, and this may result in less aggressive liver disease in HBeAg-negative CHB infection. PMID- 19811087 TI - Correlation of circulating TLR2/4 expression with CD3+/4+/8+ T cells and Treg cells in HBV-related liver cirrhosis. AB - Toll-like receptors (TLRs) 2 and 4 play a key role in chronic hepatitis B virus (HBV) infection. However, the role of TLRs in the pathogenesis of HBV-related liver cirrhosis and their regulation of the innate immune response of patients with liver cirrhosis remain unknown. To assess the contribution of TLR2/4 in HBV related liver cirrhosis, we examined the expression of circulating TLR2 and TLR4 on peripheral blood mononuclear cells (PBMCs), CD4(+)CD25(+)CD127(low/-) Treg proportions, and CD3(+)/CD4(+)/CD8(+) T-cell counts in 30 liver cirrhosis patients, 21 chronic hepatitis B (CHB) patients, and 16 normal controls (NC). Furthermore, we analyzed the relationship between TLR2/4 expression and Treg proportions and T-cell counts. We show that the expression of TLR2 and TLR4 was significantly upregulated in patients with liver cirrhosis compared to NC. TLR4 expression was significantly increased in patients with liver cirrhosis compared to patients with CHB, and for TLR2 expression there were no differences between them. TLR4 expression showed a significant positive correlation with the frequency of Tregs in liver cirrhosis patients. TLR2 expression negatively correlated with CD3(+)/CD4(+)/CD8(+) T-cell counts and HBV viral load in patients with liver cirrhosis. These findings indicate that TLR may be involved in the pathogenesis of HBV-related liver cirrhosis, and may interact with Tregs and CD3(+)/CD4(+)/CD8(+) T cells in the immune response during HBV-related liver cirrhosis. PMID- 19811088 TI - Preclinical evaluation of the immunogenicity of C-type HIV-1-based DNA and NYVAC vaccines in the Balb/C mouse model. AB - As part of a European initiative (EuroVacc), we report the design, construction, and immunogenicity of two HIV-1 vaccine candidates based on a clade C virus strain (CN54) representing the current major epidemic in Asia and parts of Africa. Open reading frames encoding an artificial 160-kDa GagPolNef (GPN) polyprotein and the external glycoprotein gp120 were fully RNA and codon optimized. A DNA vaccine (DNA-GPN and DNA-gp120, referred to as DNA-C), and a replication-deficient vaccinia virus encoding both reading frames (NYVAC-C), were assessed regarding immunogenicity in Balb/C mice. The intramuscular administration of both plasmid DNA constructs, followed by two booster DNA immunizations, induced substantial T-cell responses against both antigens as well as Env-specific antibodies. Whereas low doses of NYVAC-C failed to induce specific CTL or antibodies, high doses generated cellular as well as humoral immune responses, but these did not reach the levels seen following DNA vaccination. The most potent immune responses were detectable using prime:boost protocols, regardless of whether DNA-C or NYVAC-C was used as the priming or boosting agent. These preclinical findings revealed the immunogenic response triggered by DNA-C and its enhancement by combining it with NYVAC-C, thus complementing the macaque preclinical and human phase I clinical studies of EuroVacc. PMID- 19811089 TI - MHC expression on spleen lymphocyte subsets in genetically resistant and susceptible chickens infected with Marek's disease virus. AB - Resistance and susceptibility to Marek's disease (MD) are strongly influenced by the chicken major histocompatibility complex (MHC). In this study, splenic lymphocytes from MD-resistant and MD-susceptible chickens of three MHC genotypes (B21/B21, B19/B21, and B19/B19) were analyzed by flow cytometry for MHC surface expression. In the spleen, constitutive MHC class I surface expression was found to be highest in homozygous B19, lowest in homozygous B21, and intermediate in heterozygous B19/B21 animals. This was observed on CD4(+), CD8(+), and Bu-1(+) splenic lymphocytes. Chickens of all three genotypes were subjected to infection with MD virus (GA strain) and spleen samples from infected as well as MHC-matched negative controls were analyzed at 1, 4, and 8 wk post-infection (p.i.). It was observed that MDV induced an increase in MHC class I expression late in the infection. Thus, MHC class I was increased on the surface of CD4(+) cells from infected chickens of all genotypes at 4 and 8 wk p.i. compared with negative controls. Also, MHC class I expression was increased on CD8(+) cells from infected chickens of all genotypes at 4 and 8 wk p.i., except for the homozygous B19 animals, that showed no increase at 8 wk p.i. MDV-induced differences in MHC class II surface levels were also found. Thus, MHC class II expression was increased on CD4(+) cells from infected B19/21 and B21/B21 chickens at 4 wk p.i., and also at 8 wk p.i. on CD4(+) cells from infected B19/B21 animals. MHC class II expression was increased on CD8(+) cells from infected chickens of all genotypes at 4 wk p.i. These findings suggest that MDV infection in vivo increases the level of MHC surface expression on splenic T cells, indicating a possible role in immunity against MDV. PMID- 19811090 TI - Evaluation of the blood stabilizers TransFix and Cyto-Chex BCT for low-cost CD4 T cell methodologies. AB - TransFix(TM) and Cyto-Chex((R)) BCT (blood collection tube) reagents have been shown to maintain whole blood integrity for delayed immunophenotyping by flow cytometry. We evaluated the ability of these blood-stabilizing reagents to preserve HIV-seropositive blood for delayed CD4(+) T-cell quantification utilizing the Dynal((R)) Biotech T4 Quant Kit. TransFix was added to EDTA anticoagulated whole blood and tested at a 1:10 dilution over 7 d using the Dynal (n = 21) manual method. Compared to baseline analysis, a significant decrease in mean CD4(+) counts was observed over time. Cyto-Chex BCT-preserved samples (n = 20) were tested for CD4(+) counts by Dynal over 7 d, with storage at varying temperatures: room temperature (21 degrees C), 37 degrees C, and 37 degrees C with intermittent storage at 42 degrees C. A significant decline in mean CD4(+) counts was observed in samples at all temperatures compared to baseline (p < 0.05). Increases in temperature to and above 37 degrees C resulted in a greater decline in mean CD4(+) counts over time. Our findings indicated that neither TransFix or Cyto-Chex BCT was a suitable blood stabilizer when used for delayed CD4(+) quantification with a low-cost manual CD4(+) bead-based method. PMID- 19811091 TI - The CD94/NKG2C-expressing NK cell subset is augmented in chronic lymphocytic leukemia patients with positive human cytomegalovirus serostatus. AB - Human cytomegalovirus (HCMV) manipulates the host immune system in various ways. Allegedly, HCMV infection is associated with increased percentages of a particular natural killer (NK) cell subset expressing the activating receptor CD94/NKG2C in both healthy individuals and in patients infected with human immunodeficiency virus (HIV). Whether the HCMV-mediated induction of this specific NK cell subset is also apparent for other diseases characterized by abnormal immune responses, such as malignant blood diseases, is unknown. By comparing the fractions of CD94/NKG2C(+) NK cells in B-cell chronic lymphocytic leukemia (B-CLL) patients having either positive or negative HCMV serostatus, a proportional increase of this cell subset was obvious in the HCMV-seropositive subjects. Therapeutic intervention in the patients with positive HCMV serostatus did not seem to reduce the percentage of CD94/NKG2C-expressing NK cells. Thus, HCMV infection seemingly shapes the NK cell system in healthy individuals, HIV patients, and B-CLL patients in a uniform manner, even though these involve different immunological challenges. PMID- 19811092 TI - Tumor necrosis factor alpha enhances glutamatergic transmission onto spinal motoneurons. AB - The early stages of spinal cord injury (SCI) start with excitotoxic damage caused by a massive release of glutamate. However, glutamate release is not the only factor to consider. Inflammatory molecules like tumor necrosis factor alpha (TNFalpha), belonging to a group of cytokines initially identified and named for their ability to kill tumor cells, is also a key factor in neuronal death and inflammation. TNFalpha is released from macrophages and activated microglia following a SCI, reaching a peak 1 h after the primary injury. Motoneurons whose survival is necessary for successful rehabilitation are especially vulnerable to the effects of TNFalpha release. While TNFalpha has been postulated to increase glutamatergic synaptic transmission, evidence for this has been indirect. Here, we show using whole-cell recording from lumbar motoneurons that AMPA and NMDA receptor-mediated excitatory postsynaptic currents are rapidly increased following bath application of TNFalpha. Concurrently, the single-channel open probability of AMPA and NMDA channels were also augmented by TNFalpha. Overall, our data lead us to propose the idea that motoneuronal vulnerability to excitotoxicity is not only due to the excessive release of glutamate, but may also be attributable to the increased sensitivity of AMPARs and NMDARs to the proinflammatory factor, TNFalpha, released after SCI. PMID- 19811093 TI - Both hypoxemia and extreme hyperoxemia may be detrimental in patients with severe traumatic brain injury. AB - An association between hypoxemia and poor outcomes from traumatic brain injury (TBI) is well documented. However, it is unclear whether hyperoxygenation is beneficial. This registry-based analysis explores the relationship between early hypoxemia and hyperoxemia on outcome from moderate-to-severe TBI. TBI patients (Abbreviated Injury Scale score 3+) were identified from the San Diego County trauma registry. Patients were stratified by arrival partial oxygen pressure (Po(2)) value. Trauma and injury severity score (TRISS) was then used to calculate predicted survival for each patient, with the mean observed-predicted survival differential determined for each arrival Po(2) stratification. Logistic regression was used to quantify the relationship between hypoxemia, hyperoxemia, and outcome from TBI after adjusting for multiple variables including intubation and ventilation status. A total of 3420 patients were included in the analysis. TRISS calculations revealed worse outcomes than predicted for both hypoxemia and extreme hyperoxemia. Logistic regression revealed an optimal Po(2) range (110-487 mm Hg), with an independent association observed between decreased survival and both hypoxemia (OR 0.54; 95% CI 0.42, 0.69; p < 0.001) and extreme hyperoxemia (OR 0.50; 95% CI 0.36, 0.71; p < 0.001). The association between hypoxemia and extreme hyperoxemia and worse outcomes was also present with use of "good outcomes" as the outcome variable (discharge to home, rehabilitation, jail, or psychiatric facility, or leaving against medical advice). We conclude that both hypoxemia and extreme hyperoxemia are associated with increased mortality and a decrease in good outcomes among TBI patients. PMID- 19811094 TI - A panel of neuron-enriched proteins as markers for traumatic brain injury in humans. AB - Surrogate markers have enormous potential for contributing to the diagnosis, prognosis, and therapeutic evaluation of acute brain damage, but extensive prior study of individual candidates has not yielded a biomarker in widespread clinical practice. We hypothesize that a panel of neuron-enriched proteins measurable in cerebrospinal fluid (CSF) and blood should vastly improve clinical evaluation and therapeutic management of acute brain injuries. Previously, we developed such a panel based initially on the study of protein release from degenerating cultured neurons, and subsequently on rodent models of traumatic brain injury (TBI) and ischemia, consisting of 14-3-3beta, 14-3-3zeta, three distinct phosphoforms of neurofilament H, ubiquitin hydrolase L1, neuron-specific enolase, alpha-spectrin, and three calpain- and caspase-derived fragments of alpha-spectrin. In the present study, this panel of 11 proteins was evaluated as CSF and serum biomarkers for severe TBI in humans. By quantitative Western blotting and sandwich immunoassays, the CSF protein levels were near or below the limit of detection in pre-surgical and most normal pressure hydrocephalus (NPH) controls, but following TBI nine of the 11 were routinely elevated in CSF. Whereas different markers peaked coordinately, the time to peak varied across TBI cases from 24-96 h post-injury. In serum, TBI increased all four members of the marker panel for which sandwich immunoassays are currently available: a calpain-derived NH(2)-terminal alpha-spectrin fragment and the three neurofilament H phosphoforms. Our results identify neuron-enriched proteins that may serve as a panel of CSF and blood surrogate markers for the minimally invasive detection, management, mechanistic, and therapeutic evaluation of human TBI. PMID- 19811095 TI - Dynamic three-dimensional culture methods enhance mesenchymal stem cell properties and increase therapeutic potential. AB - Mesenchymal stem cells (MSCs) are capable of self-renewal and differentiation along the osteogenic, chondrogenic, and adipogenic lineages and have potential applications in a range of therapies. MSCs can be cultured as monolayers on tissue culture plastic, but there are indications that they lose cell-specific properties with time in vitro and so poorly reflect in vivo MSC behavior. We developed dynamic three-dimensional (3D) techniques for in vitro MSC culture using spinner flasks and a rotating wall vessel bioreactor. We characterized the two methods for dynamic 3D MSC culture and compared the properties of these cultures with monolayer MSCs. Our results showed that under optimal conditions, MSCs form compact cellular spheroids and remain viable in dynamic 3D culture. We demonstrated altered cell size and surface antigen expression together with enhanced osteogenic and adipogenic differentiation potential in MSCs from dynamic 3D conditions. By microarray analysis of monolayer and spinner flask MSCs, we identified many differences in gene expression, including those confirming widespread changes to the cellular architecture and extracellular matrix. The upregulation of interleukin 24 in dynamic 3D cultures was shown to selectively impair the viability of prostate cancer cells cultured in medium conditioned by dynamic 3D MSCs. Overall, this work suggests a novel therapeutic application for dynamic 3D MSCs and demonstrates that these methods are a viable alternative to monolayer techniques and may prove beneficial for retaining MSC properties in vitro. PMID- 19811096 TI - Roles of integrins in human induced pluripotent stem cell growth on Matrigel and vitronectin. AB - Human induced pluripotent stem cells (iPSCs) hold promise as a source of adult derived, patient-specific pluripotent cells for use in cell-based regenerative therapies. However, current methods of cell culture are tedious and expensive, and the mechanisms underlying cell proliferation are not understood. In this study, we investigated expression and function of iPSC integrin extracellular matrix receptors to better understand the molecular mechanisms of cell adhesion, survival, and proliferation. We show that iPSC lines generated using Oct-3/4, Sox 2, Nanog, and Lin-28 express a repertoire of integrins similar to that of hESCs, with prominent expression of subunits alpha5, alpha6, alphav, beta1, and beta5. Integrin function was investigated in iPSCs cultured without feeder layers on Matrigel or vitronectin, in comparison to human embryonic stem cells. beta1 integrins were required for adhesion and proliferation on Matrigel, as shown by immunological blockade experiments. On vitronectin, the integrin alphavbeta5 was required for initial attachment, but inhibition of both alphavbeta5 and beta1 was required to significantly decrease iPSC proliferation. Furthermore, iPSCs cultured on vitronectin for 9 passages retained normal karyotype, pluripotency marker expression, and capacity to differentiate in vitro. These studies suggest that vitronectin, or derivatives thereof, might substitute for Matrigel in a more defined system for iPSC culture. PMID- 19811097 TI - PCR-quality DNA isolation from human bronchial aspirates and buccal and eyelid swabs by a simple procedure based on alkaline lysis. AB - There are only a few systematic reports about DNA extraction from routine diagnostic cytological specimens. An inevitable drawback of such techniques is increased spending of time and funds required for obligatory DNA purification. To implement a simple protocol for human DNA isolation from cytological specimens related to lung cancer, bronchial aspirates together with samples collected by swabbing of the inner cheek and eyelid were used. By combining alkaline and temperature lyses it was possible to isolate DNA solution ready for PCR in less than an hour. Testing the method used for amplification of sex chromatin gene fragments showed that it is highly efficient. The presented protocol preserves high-quality DNA that is suitable for PCR-based assays. PMID- 19811098 TI - Chronic exercise reduces illness severity, decreases viral load, and results in greater anti-inflammatory effects than acute exercise during influenza infection. AB - BACKGROUND: It is assumed that moderate exercise may improve resistance to infection and reduce inflammation, but there are limited data to support this assumption in an infection model. METHODS: BALB/cJ mice were assigned to the following groups: no exercise (NON-EX), 1 session of acute exercise (A-EX), or chronic exercise for approximately 3.5 months (C-EX). Mice were infected with influenza (C-EX mice infected at rest; A-EX mice infected 15 min after exercise). RESULTS: C-EX mice demonstrated the lowest severity of infection, assessed by body weight loss and food intake. There was less virus in the lungs at day 5 after infection in C-EX and A-EX mice compared with NON-EX mice (P = .02) and less virus at day 2 after infection only in C-EX mice (P = .07). Soon after infection (day 2), interleukin 6 (IL-6), monocyte chemoattractant protein 1 (MCP 1), macrophage inflammatory protein 1beta, and tumor necrosis factor alpha in the bronchoalveolar lavage (BAL) fluid were lower in C-EX and A-EX than in NON-EX mice. At day 5 after infection, the BAL fluid from C-EX (but not A-EX) mice had less IL-6, interleukin 12p40, granulocyte colony-stimulating factor, keratinococyte-derived chemokine, and MCP-1 than that from NON-EX mice. A trend toward reduced immunopathologic response was found in C-EX mice. CONCLUSIONS: Chronic exercise resulted in reduced symptoms, virus load, and levels of inflammatory cytokine and chemokines. Acute exercise also showed some benefit, which was limited to the early phase of infection. PMID- 19811100 TI - Noninvasive real-time monitoring of liver-stage development of bioluminescent Plasmodium parasites. AB - BACKGROUND: The morbidity and mortality associated with malaria are heightened because of the spread of drug-resistant parasites and the lack of an effective vaccine. Plasmodium liver stages are the targets of new chemotherapeutics and vaccines, but there are limited tools available to study this stage in vivo. METHODS: To overcome this obstacle, we developed a method with which to study Plasmodium liver stages by means of bioluminescent imaging (BLI) of the rodent malaria parasite Plasmodium yoelii. We created a P. yoelii YM strain (PyLuc) that stably expresses firefly luciferase driven by a constitutive promoter. RESULTS: Using BLI, we performed imaging of the Plasmodium liver stages of mice infected with PyLuc sporozoites and monitored parasite dissemination during blood-stage infection. Because PyLuc luciferase activity is proportional to the number of parasites, BLI can be used to quantify the effect of drugs on liver-stage development. Moreover, using BLI, we demonstrated that immunization with blood stage parasites confers partial protective immunity against the development of liver stages. CONCLUSIONS: BLI is a noninvasive technique that is useful for screening potential drugs and candidate vaccines with which to combat malaria. The prospect of cross-stage protective immunity increases the number of avenues to be explored in the development of an effective vaccine against malaria. PMID- 19811101 TI - Hospital costs in patients with nosocomial methicillin-resistant or methicillin susceptible Staphylococcus aureus bloodstream infection. PMID- 19811099 TI - Heterogeneous vancomycin-intermediate susceptibility phenotype in bloodstream methicillin-resistant Staphylococcus aureus isolates from an international cohort of patients with infective endocarditis: prevalence, genotype, and clinical significance. AB - BACKGROUND: The significance of heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) is unknown. Using a multinational collection of isolates from methicillin-resistant S. aureus (MRSA) infective endocarditis (IE), we characterized patients with IE with and without hVISA, and we genotyped the infecting strains. METHODS: MRSA bloodstream isolates from 65 patients with definite IE from 8 countries underwent polymerase chain reaction (PCR) for 31 virulence genes, pulsed-field gel electrophoresis, and multilocus sequence typing. hVISA was defined using population analysis profiling. RESULTS: Nineteen (29.2%) of 65 MRSA IE isolates exhibited the hVISA phenotype by population analysis profiling. Isolates from Oceania and Europe were more likely to exhibit the hVISA phenotype than isolates from the United States (77.8% and 35.0% vs 13.9%; P < .001). The prevalence of hVISA was higher among isolates with a vancomycin minimum inhibitory concentration of 2 mg/L (P = .026). hVISA-infected patients were more likely to have persistent bacteremia (68.4% vs 37.0%; P = .029) and heart failure (47.4% vs 19.6%; P = .033). Mortality did not differ between hVISA- and non-hVISA-infected patients (42.1% vs 34.8%, P = .586). hVISA and non-hVISA isolates were genotypically similar. CONCLUSIONS: In these analyses, the hVISA phenotype occurred in more than one-quarter of MRSA IE isolates, was associated with certain IE complications, and varied in frequency by geographic region. PMID- 19811102 TI - Translating evidence into practice: the importance of continuing local bacterial surveillance even when national data are available. PMID- 19811104 TI - Comparison of methods of measuring pharmacy sales of antibiotics without prescriptions in Pratumthani, Thailand. PMID- 19811105 TI - Influenza pseudoinfection. PMID- 19811106 TI - Measuring hand hygiene compliance: a new frontier for improving hand hygiene. PMID- 19811107 TI - Adjuvant effect of zinc oxide on Th2 but not Th1 immune responses in mice. AB - BACKGROUND AND AIM: We investigated the effect of zinc oxide (ZnO) on Th1 and Th2 immune responses in mice. MATERIAL AND METHODS: Mice were intraperitoneally administered with ovalbumin (OVA) with or without varying doses of ZnO (day 0). On day 21, anti-OVA IgG, IgG2a, IgG1, and IgE antibodies in sera, OVA-specific proliferative responses of spleen cells, and production of Th1 cytokines including IFN-gamma as well as Th2 cytokines such as IL-4 and IL-5 were measured. RESULTS: The results showed that administration of OVA with ZnO was followed by greater increases in anti-OVA IgG and the antigen-specific splenocyte proliferation compared to that of OVA alone. The production of anti-OVA IgG1 and IgE and secretion of IL-4 and IL-5 were markedly enhanced by ZnO. The enhancing effect of ZnO on these Th2 responses was as strong as aluminium hydroxide (Alum) that was widely used as an adjuvant. In contrast, treatment with OVA plus ZnO failed to affect production of anti-OVA IgG2a as well as IFN-gamma. It was also observed that ZnO had a stimulating effect on the secretion of the proinflammatory cytokine IL-17 from a new lineage of effector Th cells. CONCLUSION: These results suggest that ZnO appears to have an adjuvant effect on the immune system, especially Th2 but not Th1 immune responses. PMID- 19811108 TI - Methanol extract of Phellodendri cortex alleviates lipopolysaccharide-induced acute airway inflammation in mice. AB - BACKGROUND AND AIM: The effects of methanol extract of Phellodendri cortex on acute airway inflammation induced by intranasal administration of lipopolysaccharide (LPS, 300mug/kg) were investigated in female BALB/c mice. MATERIALS AND METHODS: At 2 h after LPS exposure, mice were treated orally with methanol extract of Phellodendri cortex (100, 200 and 400 mg/kg). At the end of this study, bronchoalveolar lavage fluids (BALF) were collected and number of total cells, macrophages and neutrophils, protein concentration were analyzed. Tumor necrosis factor-alpha (TNF-alpha), macrophage inflammatory protein (MIP-2), IL-10 levels and nitric oxide (NO) production in BALF were also determined. RESULTS: Methanol extract of Phellodendri cortex dose-dependently alleviated LPS induced acute airway inflammation via decreasing the infiltration of inflammatory cells and the release of inflammatory mediators. CONCLUSION: The relief of airway inflammation provides a possible therapeutic application of Phellodendri cortex for the treatment of infectious pulmonary diseases. PMID- 19811109 TI - Examining and interpreting responsiveness of the Diabetes Medication Satisfaction measure. AB - OBJECTIVE: Treatment satisfaction (TS) is an important patient reported outcome (PRO) in diabetes as it is correlated with outcomes necessary for optimal treatment (e.g., compliance, self-management behaviour). The objective of this study was to examine the responsiveness of the DiabMedSat, a disease-specific PRO measure, assessing Overall, Burden, Efficacy and Symptom TS. METHODS: The DiabMedSat was included in an open label, observational study of the safety and efficacy of biphasic insulin aspart 30 (NovoMix 30) in routine practice with type 2 diabetes. Responsiveness analyses, examining both internal and external responsiveness, were conducted and minimally important differences (MID) assessed. RESULTS: In 18,817 patients, all TS scores significantly improved after 26 weeks of treatment (p<0.001). The effect sizes for these changes were above 0.5 indicating that the ability to detect change was moderate-to-large in size. Significant differences were found for all TS scores comparing patients who met their HbA(1c) goal, who improved but did not meet goal and who did not improve (p<0.01), and for patients who experienced a minor hypoglycaemic event and those who did not (p<0.001). DiabMedSat scores were able to detect changes in patients' own global rating of satisfaction (MID ranging from 5.3 to 11.7) and in physician rated satisfaction with patients' HbA(1c) improvement (MID ranging from 5.3 to 10.2). CONCLUSIONS: In the context of an observational study, the DiabMedSat has been shown to be highly responsive to change and can be considered as an acceptable PRO measure for TS in diabetes. PMID- 19811110 TI - Cost effectiveness of palivizumab in children with congenital heart disease in Germany. AB - OBJECTIVES: To assess the cost effectiveness of palivizumab, a humanised monoclonal antibody, used as prevention against severe respiratory syncytial virus (RSV) infection requiring hospitalisation, in infants with haemodynamically significant congenital heart disease (CHD) in the German healthcare setting. STUDY DESIGN: A decision-tree model was used to estimate the cost effectiveness of palivizumab for a hypothetical cohort of patients. The analysis was based on a lifetime follow-up period in order to capture the impact of palivizumab on long term morbidity and mortality resulting from an RSV infection. Data sources included published literature, the palivizumab pivotal trials, official price/tariff lists and national population statistics. The study was conducted from the perspective of society (primary analysis) and the healthcare purchaser (secondary analysis). RESULTS: From the societal perspective, use of palivizumab results in an incremental cost-effectiveness ratio (ICER) of ?2,615 per quality adjusted life-year (QALY) without discounting, which increases to ?9,529/QALY after discounting. From the perspective of the German healthcare purchaser, use of palivizumab results in an ICER of ?4,576/QALY without discounting, which increases to ?16,673/QALY after discounting. Probabilistic sensitivity analyses confirmed the robustness of the model. The study is limited by a number of conservative assumptions. It was assumed that palivizumab only affects the occurrence of RSV hospitalisation and does not influence the severity of the RSV infection. Another assumption was that international clinical trial data and data on utilities could be applied to the German healthcare setting. CONCLUSION: This analysis showed that palivizumab represents a cost-effective means of prophylaxis against severe RSV infection requiring hospitalisation in infants with haemodynamically significant CHD. PMID- 19811111 TI - Cost effectiveness of palivizumab for RSV prevention in high-risk children in the Netherlands. AB - BACKGROUND: Respiratory syncytial virus (RSV) is a common pathogen that is the leading cause of lower respiratory tract infections in young children. High-risk children are at risk of severe infection, which may require hospitalisation. RSV is also associated with a high risk for respiratory morbidity and mortality, which may have long-term clinical and economic consequences. OBJECTIVE: To assess the cost effectiveness of palivizumab, a humanised monoclonal antibody, used as prevention against severe respiratory syncytial virus (RSV) infection requiring hospitalisation, in the indication of preterm infants and infants with preterm/bronchopulmonary dysplasia and in the second indication of children with congenital heart disease in the Dutch healthcare setting. METHODS: A decision tree model was used to estimate the cost effectiveness of palivizumab, used as a preventative treatment against severe respiratory syncytial virus (RSV) infection, in high-risk groups of children in the Netherlands. The analysis was based on a lifetime follow-up period in order to capture the impact of palivizumab on long-term morbidity and mortality resulting from an RSV infection. Data sources included published literature, the palivizumab pivotal trials, official price/tariff lists and national population statistics. The study was conducted from the perspective of society in the Netherlands. RESULTS: The use of palivizumab results in undiscounted incremental cost-effectiveness ratios of ?12,728/QALY and ?4,256/QALY in preterm/bronchopulmonary dysplasia and congenital heart disease indications, respectively. Inclusion of indirect costs leads to even more favourable cost-effectiveness outcomes. The study is limited by a number of conservative assumptions. It was assumed that palivizumab only affects the occurrence of RSV hospitalisation and does not influence the severity of the RSV infection. Another assumption was that international clinical trial data and data on utilities could be applied to the Dutch healthcare setting. CONCLUSION: Palivizumab provides cost-effective prophylaxis against RSV in high-risk infants. The use of palivizumab in these children results in positive short- and long-term health-economic benefits. PMID- 19811112 TI - Clinical study of the effects on asthma-related QOL and asthma management of a medical food in adult asthma patients. AB - BACKGROUND: Asthma can have a negative impact on quality of life although this is not well correlated with objective evaluations of pulmonary function. A medical food, EFF1009, containing the fatty acids gamma-linolenic acid (GLA) and eicosapentaenoic acid (EPA) decreases leukotriene B(4) synthesis in patients with asthma. Two previous clinical studies with EFF1009 provided preliminary evidence that the medical food improves asthma-related quality of life (ARQOL) and asthma management. OBJECTIVE: To evaluate the impact on ARQOL of EFF1009 in adults with asthma. RESEARCH DESIGN AND METHODS: The study was a randomized, prospective, double-blind, placebo-controlled, parallel group study in twenty-one (N = 21 evaluable) subjects with mild to moderate persistent asthma who consumed the medical food emulsion or placebo emulsion daily for 28 days. All participants continued their asthma medications throughout the study. ARQOL, including asthma signs and symptoms, and asthma control were measured using the Mini Asthma Quality of Life Questionnaire (MiniAQLQ) and the Asthma Control Questionnaire (ACQ), administered at baseline, Day 14 and Day 28. Safety and tolerability parameters, including adverse events, were monitored. RESULTS: Baseline ARQOL scores, forced expiratory volume in one second (FEV(1)) and other characteristics were balanced between both groups. Mean (standard error) total MiniAQLQ scores changed by 0.73 (0.38) and -0.22 (0.36) in the EFF1009 and placebo groups, respectively, (p < 0.05). The MiniAQLQ symptom domain score was improved in the EFF1009 group (p < 0.05). Total scores for the ACQ were not significantly improved in either group. Levels of the fatty acid EPA in plasma increased in the EFF1009 group but not the placebo group (p < 0.03). The medical food was well tolerated and no safety concerns were identified. CONCLUSIONS: The dietary addition of the medical food EFF1009 to asthma management regimens can improve patient perceived, ARQOL and can also improve asthma management as evidenced by reduced asthma symptoms. An additional study of the medical food, with larger subject population and longer treatment duration, is warranted to confirm these findings. PMID- 19811113 TI - Biomarkers of cellular reaction to pesticide exposure in a rural population. AB - In the present study we report data obtained from the evaluation of subjects occupationally exposed to pesticide mixtures from Santa Fe province, Argentina, using biomarkers for butyrylcholinesterase (BChE) and acetylcholinesterase (AChE) activities, catalase (CAT), lipid peroxidation (by TBARS assay) and the Damage Index Comet Assay (DICA). Our results showed an AChE inhibition (25% and 15% in directly and indirectly groups, respectively) in relation to controls with no significant modifications in BChE. TBARS levels were higher (51%) in pesticide sprayers while CAT activity was reduced in both, applicators (61%) and non applicators (43%). DICA was significantly increased in direct (83%) and indirect (98%) exposed groups, compared with controls. These results showed modifications in lipid peroxidation, antioxidant defence system, and DNA damage in lymphocytes of exposed workers. Further investigations are suggested in order to link our findings with adverse health effects observed in chronic pesticide toxicity, where oxidative damage plays a pathophysiological role. PMID- 19811114 TI - National, European licensing examinations or none at all? PMID- 19811115 TI - Online eAssessment: AMEE guide no. 39. AB - In this guide, the authors outline the advantages of online eAssessment and examine the intellectual, technical, legal and cost issues that arise from its use. This guide outlines the major assessment types that are suitable for online assessment and makes a key distinction between formative and summative assessment. The focus is primarily on the latter since that is where the difficulties are most acute and robust systems most critical. A range of practical issues relating to the key stages in running a summative e-exam are explored and advice given on system requirements and on how to ensure that the exam runs smoothly when you 'go live'. This section includes consideration of the way that using eAssessment might affect the standard setting and results analysis process. The section on future trends in online assessment explores possibilities such as computer adaptive testing and the automated assessment of free text answers. Finally, there is a consideration of the implications of these trends for management. PMID- 19811116 TI - Asking the right questions and getting meaningful responses: 12 tips on developing and administering a questionnaire survey for healthcare professionals. AB - Questionnaires provide a useful and versatile tool for new and occasional researchers, and can be applied to a wide range of topics. This paper provides simple guidance on some of the potential pitfalls in developing and running a questionnaire study, and how to avoid them. Each tip is illustrated with a real life example from the development of a UK-wide questionnaire survey of trainee doctors and their educational supervisors. PMID- 19811117 TI - Licensing examinations in North America: is external audit valuable? AB - The United States and Canada both have long-standing, highly developed national systems of assessment for medical-licensure based outside the institutions of medical education. This commentary reviews those programs and explores some of the reasons for their implementation and retention for nearly a century. The North American experience may be relevant to dialog about national or European assessments for medical practice. PMID- 19811118 TI - European licensing examinations--the only way forward. PMID- 19811119 TI - Five myths and the case against a European or national licensing examination. AB - The introduction of a European licensing examination or national examinations, where these do not already exist, offers significant advantages. These are more than offset, however, by the disadvantages and the collateral damage incurred. Five myths about centralizing examinations are explored. Myth 1: The claim that a central examination will ensure that candidates are assessed in important areas of medical practice is unfounded. What tends to be assessed are learning outcomes that can be easily assessed. These are often not the important outcomes related to the overall competence of a doctor. Myth 2: It is claimed that a central examination will lead to improvements in assessment practice. The evidence is that this is not the case and that, in fact, a central examination stifles change and inhibits innovation. Myth 3: A central examination, it is suggested, will meet a need for greater uniformity. There is also an important need to recognize diversity. Myth 4: Central examinations are seen as an indicator that will track the performance of the system. The limitations of the data, however, are usually not recognized and there maybe unfortunate and unintended consequences if the results are used in this way. Myth 5: Finally, a major argument proposed for a European or national examination is that it will lead to safer medical practice and that this will protect the patient from substandard practitioners. There is, in fact, no evidence to support this argument. There is a need for further work and new initiatives on standards and quality improvement in assessment approaches. This can be achieved in a number of ways including monitoring the assessment process and sharing tools and assessment approaches between schools. PMID- 19811120 TI - The utility of (European) licensing examinations. AMEE Symposium, Prague 2008. PMID- 19811122 TI - Radiology: an underutilized resource for undergraduate curricula. PMID- 19811123 TI - Undergraduate research internships: veterinary students' experiences and the relation with internship quality. AB - The learning environment of undergraduate research internships has received little attention, compared to postgraduate research training. This study investigates students' experiences with research internships, particularly the quality of supervision, development of research skills, the intellectual and social climate, infrastructure support, and the clarity of goals and the relationship between the experiences and the quality of students' research reports and their overall satisfaction with internships. METHOD: A questionnaire (23 items, a 5-point Likert scale) was administered to 101 Year five veterinary students after completion of a research internship. Multiple linear regression analyses were conducted with quality of supervision, development of research skills, climate, infrastructure and clarity of goals as independent variables and the quality of students' research reports and students' overall satisfaction as dependent variables. RESULTS: The response rate was 79.2%. Students' experiences are generally positive. Students' experiences with the intellectual and social climate are significantly correlated with the quality of research reports whilst the quality of supervision is significantly correlated with both the quality of research reports and students' overall satisfaction with the internship. CONCLUSION: Both the quality of supervision and the climate are found to be crucial factors in students' research learning and satisfaction with the internship. PMID- 19811124 TI - I came, I saw, I reflected: a qualitative study into learning outcomes of international electives for Japanese and British medical students. AB - BACKGROUND AND AIM: Although medical students have increasingly more opportunities to participate in international electives, their experiences are usually unstructured and the literature referring to their learning outcomes, educational environment, and assessment is scanty. This study was undertaken to clarify qualitatively what students learn from their international electives. METHODS: We carried out semi-structured individual interviews with 15 Japanese students studying clinical medicine in British medical schools and six British students studying in Japanese medical schools. The thematic synthesis method was used in analysing the transcribed data and triangulation by multiple researchers was used to achieve higher reliability. RESULTS: The main learning outcomes identified were skills in history taking and physical examination with clinical reasoning and in management of diseases rarely seen in the students' own countries; awareness of clinical ethics and merits and demerits of different systems of healthcare and medical education; sensitivity to issues in doctor patient relationships and work ethics; enhancement of cultural competence; and personal development. CONCLUSIONS: Most learning outcomes of international electives are culture- or system-dependent. Students achieved outcomes related closely to medical professionalism, mainly through reflection. International electives may give students opportunities to learn both professionalism and cultural competence. PMID- 19811125 TI - Large group high-fidelity simulation enhances medical student learning. AB - BACKGROUND: Previous work shows feasibility for large group high-fidelity simulation with correlation to basic science in the preclinical curriculum. AIMS: This project studies whether large group simulation leads to enhanced basic science learning. METHODS: This was an educational performance study before and after high-fidelity simulation for first-year medical students. Basic neuroscience concepts were reinforced with simulation, and pretesting and posttesting were analysed along with summative exam results. The number correct was compared on a contingency table using the Mantel-Haenszel chi-squared test and same student correlation was accounted for with a 'Generalized Estimating Equations' model. RESULTS: The study included 112 students; three were excluded for missing data. Students showed statistically significant improvement on two of the four questions, and a nonsignificant improvement or equivalent performance on two questions. Students were significantly more likely to get all four responses correct on the posttest than on the pretest. Summative testing 11 days later had >80% correct responses for three factual recall questions and 58% correct responses for a single knowledge application question. CONCLUSIONS: Simulation is an effective teaching method for preclinical basic science education. Students demonstrated significant improvements after participating in a live interactive simulation scenario. PMID- 19811126 TI - SOMOSAT: Utility of a web-based self-assessment tool in undergraduate medical education. AB - BACKGROUND: Relatively few studies have rigorously assessed the effectiveness of computer-based self-assessment in medical education. AIM: To assess whether an online self-assessment tool can be an effective adjunct to a traditional curriculum for second-year medical students. METHODS: The NYU School of Medicine Online Self-Assessment Tool (SOMOSAT) consists of >450 multiple-choice questions spanning disciplines of internal medicine, administered as separate modules focused on individual organ systems. Questions are coded on multiple dimensions, permitting second-year medical students to receive low-stakes, highly specific feedback regarding their knowledge and performance. Students can also review their answers to guide future study. We employed data collected during SOMOSAT operation to assess its utility and effectiveness. RESULTS: Overall, SOMOSAT accurately predicted student performance on future exams. SOMOSAT participants generally performed better than non-participants on subsequent graded course examinations (p < 0.05). Students using SOMOSAT subsequently experienced greater improvement in areas in which they initially performed poorly, compared with those in which they initially performed well. Students reported that SOMOSAT was most helpful in filling knowledge gaps, and providing opportunities to practice exam-style questions. CONCLUSION: The ability of SOMOSAT to enhance learning and exam performance suggests that web-based self-assessment tools can be effective adjuncts to traditional educational methods. PMID- 19811127 TI - Mapping the teacher's role: the value of defining core competencies for teaching. PMID- 19811128 TI - The place of anatomy in medical education: AMEE Guide no 41. AB - This Guide, a combined work by three authors from different countries, provides perspectives into the history of teaching gross anatomy, briefly, from the earliest of times, through to a detailed examination of curricula in both traditional didactic approaches and Problem-Based Learning (PBL) curricula. The delivery of a module within a curriculum in tertiary education is interplay between the content (knowledge and skills) of a subject, the teaching staff involved, the students and their approaches to learning, and the philosophy underpinning the delivery of the learning material. The work is divided into sections that deal with approaches to learning anatomy from the perspective of students, to delivery of the content of the curriculum by lecturers, including the assessment of knowledge, and itemises the topics that could be considered important for an appropriate anatomy module in an integrated course, delivered in a way that emphasises clinical application. The work concludes by looking to the future, and considering what measures may need to be addressed to ensure the continued development of anatomy as a clinically relevant subject in any medical curriculum. PMID- 19811129 TI - A framework of teaching competencies across the medical education continuum. AB - BACKGROUND: The quality of teachers in higher education is subject of increasing attention, as exemplified by the development and implementation of guidelines for teacher qualifications at Universities in The Netherlands. AIM: Because medical education takes a special position in higher education the Council of Deans of Medical Schools in The Netherlands installed a national task force to explore a method to weigh criteria for teacher qualifications of medical teachers. METHODS: A framework was developed covering competencies of teachers throughout the medical education continuum and including medicine, dentistry and veterinary medicine. RESULTS: The framework distinguishes 3 dimensions: (a) six domains of teaching (development - organization - execution - coaching - assessment - evaluation); (b) three levels in the organization at which teachers perform (micro, meso and macro level) and (c) competencies as integration of knowledge, skills and attitude and described as behaviour in specific context. The current framework is the result of several cycles of descriptions, feedback from the field and adaptations. It is meant as a guideline, leaving room for local detailing. CONCLUSION: The framework provides a common language that may be used not only by teachers and teacher trainers, but also by quality assurance committees, human resource managers and institutional boards. PMID- 19811130 TI - Medical interns' view of their undergraduate medical education in Uppsala: an alumnus study with clear attitude differences between women and men. AB - BACKGROUND: An alumni study of graduates from the medical school in Uppsala, was performed to give input into an ongoing reform process. AIMS: This study aimed to investigate how medical interns view their undergraduate medical education and the extent to which they felt that the curriculum prepared them for their current positions. METHODS: A web-based questionnaire was sent out via mail in 2005 to all past graduates who had qualified in Uppsala in 2003. RESULTS: Replies were obtained from 69 of 102 students (68%). The most apparent suggested change of the education was increased integration of preclinical and clinical teaching. Correlations were found between student satisfaction with the medical school and perceived teacher attitude, encouragement to reflect, and the graduates' perception of having sufficient practical abilities. Significant gender differences were found regarding perceived clinical ability and concerning feedback and encouragement from the teachers. CONCLUSIONS: Our results suggest more direct feedback from the teachers and more integration between basic sciences and clinical education. Female and male students may have different needs. A key question is therefore to encourage teachers to learn about gender since female and male students should equally experience respectful encounters with teachers and doctors acting as role models. PMID- 19811131 TI - What is a good doctor? The impact of 'fruitful irrelevance' in medical education. AB - Medicine is a science and evidence-based profession. Therefore, medical education and training should offer optimal conditions to help students to become a medical expert. However, in order to become a 'good doctor' the development of additional skills should also be stimulated. By including subjects like philosophy of science, medical history and sociology, the link between literature and medicine, and other 'unrelated and irrelevant' academic subjects, medical education programmes can stimulate that future doctors will broaden their mind. This will contribute to their professional performance. PMID- 19811132 TI - e-Learning in medical education: Guide supplement 32.5--Viewpoint. PMID- 19811133 TI - e-Learning in medical education: Guide supplement 32.6--Practical application. PMID- 19811134 TI - Needs assessment for continuing medical education in Nepal. PMID- 19811135 TI - Teaching psychiatric diagnostics to general practitioners: Educational methods and their perceived efficacy. AB - INTRODUCTION: Psychiatric disorders are hard to detect in a primary care setting. The vocational training for general practitioners (GPs) of the University of Maastricht, the Netherlands, intends to create a new comprehensive programme on diagnosing psychiatric disorders. AIM: We consulted the literature to obtain an answer to the question: is evidence available for the effectiveness of specific educational methods to teach GP trainees psychiatric diagnostic skills? METHODOLOGY: We searched in four databases for studies on a variety of search terms (39) referring to primary care, psychiatry, diagnosis, education and quality. As selection criterium for outcome measure we took change in diagnostic competence. RESULTS: From a number of 769 articles 27 methodologically sound studies remained. This article presents several of their research characteristics. No conclusive evidence has been found for the effectivity of an isolated educational method. The combination of methods seems promising. However, no specific mix of methods is a guarantee for success. CONCLUSIONS: We made some recommendations for training psychiatric diagnostic competency. The literature endorses our own idea that education in this field should include reflection on attitude and barriers. PMID- 19811136 TI - The art and science of debriefing in simulation: Ideal and practice. AB - OBJECTIVES: Describing what simulation centre leaders see as the ideal debriefing for different simulator courses (medical vs. crisis resource management (CRM) oriented). Describing the practice of debriefing based on interactions between instructors and training participants. METHODS: Study 1 - Electronic questionnaire on the relevance of different roles of the medical teacher for debriefing (facilitator, role model, information provider, assessor, planner, resource developer) sent to simulation centre leaders. Study 2 - Observation study using a paper-and-pencil tool to code interactions during debriefings in simulation courses for CRM for content (medical vs. CRM-oriented) and type (question vs. utterance). RESULTS: Study 1 - The different roles were seen as equally important for both course types with the exception of 'information provider' which was seen as more relevant for medical courses. Study 2 - There were different interaction patterns during debriefings: line - involving mostly the instructor and one course participant, triangle - instructor and two participants, fan - instructor and all participants in a dyadic form and net - all participants and the instructor with cross references. CONCLUSION: What simulation centre heads think is important for the role mix of simulation instructors is (at least partly) not reflected in debriefing practice. PMID- 19811137 TI - Shaping professionalism in pre-clinical medical students: Professionalism and the practice of medicine. AB - BACKGROUND: Increasing emphasis is placed on teaching and assessment of professionalism in the continuum of medical education. Consistent and longitudinal instruction and assessment are crucial factors that learners need in order to internalize the tenets of professionalism. AIM: We aimed to develop a novel longitudinal course in professionalism spanning the first 2 years in a medical curriculum. METHODS: This is a description of the process undertaken over the past 7 years to develop and implement a professionalism curriculum. We used the conceptual framework of constructivism, principles of adult learning, experiential learning and reflective practice to integrate learning with experience. We included student input in session development. Faculty mentors serve as role models to guide, assist and counsel students. Assessment of learners is accomplished using self, peer and mentor evaluation, and a student portfolio. Program evaluation is by course and faculty evaluation. RESULTS: Students are given a final grade of pass or fail, together with a brief narrative. Course evaluations were positive. A survey questionnaire showed that more than 60% of the students reported gaining skills related to course goals. CONCLUSIONS: A longitudinal curriculum for the pre-clinical years was successfully launched. Plans are under way to expand this into the clinical years. PMID- 19811138 TI - Influence of educational programs on attitudes of medical students towards psychiatry: Effects of psychiatric experience, gender, and personality dimensions. AB - BACKGROUND: Attitudes of medical students form the basis for medical actions. Because of the specific characteristics of psychiatric patients, positive attitudes of medical students towards psychiatry should be a higher goal in medical education. AIM: We hypothesize that medical students in different educational programs develop different attitudes towards psychiatry. METHODS: In a cross-sectional study, students enrolled in different educational programs completed the 'attitudes towards psychiatry' questionnaire (ATP-30). Data concerning experiences in psychiatry, personality traits and socio-demographic variables including gender were also analyzed. RESULTS: The response rate of students in the PBL-curriculum (n = 61) was >90%, in the traditional curriculum (n = 280) >75%. Attitudes towards psychiatry of male students in the Problem Based Learning program were equal to the female students' attitudes in both programs. Female students' attitudes in the traditional curriculum reached comparably good results while male students' displayed the worst attitudes. The personality factors 'openness to experience' and 'agreeableness' correlated significantly with positive attitudes towards psychiatry. PBL-students showed significantly more 'openness to experience'. CONCLUSION: Educational programs might play a role for the development of attitudes towards psychiatry, especially in male students. Factors influencing enrollment into special educational programs should also have been taken into account. An independent study with a larger number of participants will be required to support these findings. PMID- 19811139 TI - Are medical students agreeable? An exploration of personality in relation to clinical skills training. AB - BACKGROUND: Clinical competencies like trust, empathy, and cooperation are emphasized in medical school curricula. Agreeableness, a personality domain, reflects these competencies. It is unclear, however, whether medical student personality is intrinsically agreeable. AIM: We explored whether medical student personality reflects Agreeableness, and compared student Agreeableness with that of police officer recruits, a group in which high Agreeableness is not preferred. METHODS: Students and recruits completed the Revised NEO Personality Inventory, which measures domains of the five-factor model: Neuroticism, Extraversion, Openness, Agreeableness, and Conscientiousness. RESULTS: Medical student Agreeableness was at average levels. Students were high in Extraversion and Openness, reflecting personal growth, leadership, problem solving, and influencing. Relative to recruits, students had higher Neuroticism and Openness and lower Conscientiousness. Agreeableness and Extraversion did not differ. Using discriminant analysis, Neuroticism, Openness, and Conscientiousness accurately classified 77% of students and recruits. CONCLUSION: Medical students were not inordinately agreeable. They were ambitious, intellectually-creative problem solvers with a preference to direct/influence. Clinical skills training that acknowledges this style may enhance clinical education processes. Model-based methods for clinical skills--including agenda-setting, conflict resolution, and alliance making - that require mastery of techniques and have evidence-based relevance to patient care may be useful adjuncts to conventional clinical training. PMID- 19811140 TI - Clinical supervisors' perceived needs for teaching communication skills in clinical practice. AB - BACKGROUND: Lack of faculty training is often cited as the main obstacle to post graduate teaching in communication skills. AIMS: To explore clinical supervisors' needs and perceptions regarding their role as communication skills trainers. METHODS: Four focus group discussions were conducted with clinical supervisors from two in-patient and one out-patient medical services from the Geneva University Hospitals. Focus groups were audio taped, transcribed verbatim and analyzed in a thematic way using Maxqda software for qualitative data analysis. RESULTS: Clinical supervisors said that they frequently addressed communication issues with residents but tended to intervene as rescuers, clinicians or coaches rather than as formal instructors. They felt their own training did not prepare them to teach communication skills. Other barriers to teach communication skills include lack of time, competing demands, lack of interest and experience on the part of residents, and lack of institutional priority given to communication issues. Respondents expressed a desire for experiential and reflective training in a work-based setting and emphasised the need for a non-judgmental learning atmosphere. CONCLUSIONS: Results suggest that organisational priorities, culture and climate strongly influence the degree to which clinical supervisors may feel comfortable to teach communication skills to residents. Attention must be given to these contextual factors in the development of an effective communication skills teaching program for clinical supervisors. PMID- 19811141 TI - The influence of early clinical experiences on career preference of male and female medical students. AB - BACKGROUND: Clinical experience is considered to affect medical students' career preferences. It is not known whether the sequence of the clinical rotations influences these preferences. AIM: To explore whether the first clinical clerkship has more impact on career preference than the second by examining the association between the first clinical clerkship and the choice of an elective sixth-year internship. METHOD: University Medical Center Utrecht students are assigned to either a surgical or a medical ward for the first third-year clerkship and to the other ward for the second clerkship. In a retrospective cohort study, internship data of 488 sixth-year students were related to their first clerkship 3 years earlier. RESULTS: For the group as a whole, no association was found between third-year clerkship and sixth-year internship. However, male students who had been assigned to surgery first more often chose a surgical internship than those who had been assigned to medical clerkship first and vice versa (p < 0.02). Within the female subgroup, no association was found. CONCLUSION: A positive association between the nature of the clerkship and the sixth-year internship preference among male students suggest that the first clinical experience can affect later specialty preference. PMID- 19811142 TI - Cinemeducation: A pilot student project using movies to help students learn medical professionalism. AB - BACKGROUND: Using movies has been accepted worldwide as a tool to help students learn medical professionalism. In the second year, a group of medical students conducted the "Cinemeducation" project to promote professionalism in the "Medical Ethics and Critical Thinking" course. METHOD: Five movies with professionalism issues were screened with 20-30 students attending each session. After the show, participants then were asked to reflect on what they had learned in terms of professionalism. Two students led group discussion emphasizing questioning and argumentation for 60 min. Additional learning issues emerging from each session were also explored in more depth and arranged into a report. RESULTS: In the Cinemeducation Project, medical students have learned five main ethical issues in each film, which were the doctor-patient relationship, informed consent and clinical trials in patients, management of genetic disorders, patient management, and brain death and organ transplantation. In addition to issues of professionalism, they also developed critical thinking and moral reasoning skills. CONCLUSION: Using a case-based scenario in movies has proven to be an effective and entertaining method of facilitating students with learning on professionalism. PMID- 19811143 TI - Scholarship, publication, and career advancement in health professions education: AMEE Guide No. 43. AB - Scholarship and publication are key contributors to career advancement in health professions education worldwide. Scholarship is expressed in many ways including original research; integration and synthesis of ideas and data, often across disciplines; application of skill and knowledge to problems that have consequences for health professionals, students, and patients; and teaching in many forms. Professional publication also has diverse outlets ranging from empirical articles in peer reviewed journals, textbook chapters, videos, simulation technologies, and many other means of expression. Scholarship and publication are evaluated and judged using criteria that are consensual, public, and transparent. This three-part AMEE Guide presents advice about how to prepare and publish health professions education research reports and other forms of scholarship in professional journals and other outlets. Part One addresses scholarship-its varieties, assessment, and attributes of productive scholars and scholarly teams. Part Two maps the road to publication, beginning with what's important and reportable and moving to manuscript planning and writing, gauging manuscript quality, manuscript submission and review, and writing in English. Part Three offers 21 practical suggestions about how to advance a successful and satisfying career in the academic health professions. Concluding remarks encourage health professions educators to pursue scholarship with vision and reflection. PMID- 19811144 TI - Medical education in Germany. AB - Following the changes made to the medical licensing regulations of 2002, medical education in Germany has been subject to radical modification, especially at undergraduate level. The implementation of the Bologna Process is still a matter of intense political debate, whilst positive movement has occurred in developing the professionalisation of teaching staff through a Masters Degree in Medical Education. In the area of postgraduate medical education, major restructuring of programmes is occurring, whilst the debate in continuing medical education is related to the amount of practical clinical education that is required. PMID- 19811145 TI - Twelve tips for studying medical education at doctoral level. AB - BACKGROUND: Medical educators increasingly need to develop their research skills to produce robust medical education research, resulting in publication. This need has fueled rising enrollment of medical educators on doctoral programmes and the proliferation of courses, each with its own strengths and weaknesses which need to be considered before making a commitment to a long period of study. AIMS: We aimed to provide advice about the important steps in studying for a doctorate in medical education, from the decision to undertake the doctorate, through the practicalities of engaging supervisors and choosing research questions, to what to do after the doctorate is completed. METHOD: Critical reflection upon our experiences of studying for each type of medical education doctorate (the PhD, the MD and the EdD), combined with evidence from the literature where available. RESULTS AND CONCLUSIONS: The journey to achieving a doctorate in medical education can be long and bumpy with periods of disbelief and despondency. By being realistic and honest with oneself at the outset, analysing one's motivations, deciding which of the different types of doctorate best suits one's needs, finding an appropriate supervisor, creating a researchable question and getting the right support both from one's supervisor and your peers, the journey can be made manageable and even enjoyable. PMID- 19811146 TI - Maturity and medical students' ease of transition into the clinical environment. AB - BACKGROUND: Medical education has been characterized in terms of points of transition, which are accentuated by lack of relevant prior experience and can lead to extreme positive and negative emotions. AIMS: Quantify the effect of maturity on medical students' transitions into the clinical environment and identify how experiences of transition might be improved. METHOD: Eleven weeks after entering the clinical environment, 29 mature students (age over 21 at entry, median age 22) in a horizontally-integrated, predominantly undergraduate entry, problem-based curriculum offering little early clinical exposure and 58 matched non-mature students (median age 18 years) rated their experiences of transition and wrote free text comments about them. RESULTS: 62% of mature students compared with 24% of controls described 'good transitions' (odds ratio [OR] for a good transition 6.1; p = 0.002) and mature students were more likely than controls to describe how they drew on their previous years in medical school (OR 2.7, p = 0.04) and their wider life experiences in making the transition (OR 3.9, p = 0.01). They were less likely to feel confused or daunted. Whether mature or not, prior workplace experience, having learned the theory of medicine by PBL, and being confident in their knowledge and skills helped students' transitions. Both mature and non-mature students valued the support of teachers and peers and would have valued clinical experience earlier. CONCLUSIONS: The fact that just a few extra years of life experience made such a large difference to students' experiences of transition illustrates how important social factors are in the personal development of medical students. In respondents' views, early clinical experience and early skills training could ease students' passage into the clerkship phase of their education. PMID- 19811147 TI - Medical student involvement in website development. AB - BACKGROUND: The digital management of educational resources and information is becoming an important part of medical education. AIMS: At Vanderbilt University School of Medicine, two medical students sought to create a website for all medical students to act as each student's individual homepage. METHOD: Using widely available software and database technology, a highly customized Web portal, known as the VMS Portal, was created for medical students. Access to course material, evaluations, academic information, and community assets were customized for individual users. Modular features were added over the course of a year in response to student requests, monitoring of usage habits, and solicitation of direct student feedback. RESULTS: During the first 742 days of the VMS Portal's release, there were 209,460 student login sessions (282 average daily). Of 348 medical students surveyed (71% response rate), 84% agreed or strongly agreed that 'consolidated student resources made their lives easier' and 82% agreed or strongly agreed that their needs were represented by having medical students design and create the VMS Portal. CONCLUSION: In the VMS Portal project, medical students were uniquely positioned to help consolidate, integrate, and develop Web resources for peers. As other medical schools create and expand digital resources, the valuable input and perspective of medical students should be solicited. PMID- 19811148 TI - Medical faculty and curriculum design - 'No, no, it's like this: You give your lectures ...'. AB - BACKGROUND AND AIMS: The purpose of this study was to understand more completely the (tacit) curriculum design models of medical faculty. We report on two research questions: (1) Can medical faculty give an account of their curriculum design assumptions? and (2) What are their assumptions concerning curriculum design? METHOD: We conducted an explorative, qualitative case study. We interviewed educational decision makers at the three Danish medical schools and associate professors from different courses concerning curriculum design. We carried out four individual, in-depth interviews and four focus groups with 20 participants in all. RESULTS AND CONCLUSIONS: Only one decision maker had an explicit curriculum design model. However, all participants had assumptions concerning curriculum design. We displayed their assumptions as five essentially different and increasingly complex models: the method-driven, pragmatically driven content-driven, outcome-driven and vision-driven curriculum design models. In the five models, the role of learning outcomes differs. The differences range from a belief that learning outcomes are essential, to a belief that learning outcomes are unimportant, to a belief that learning outcomes are incompatible with higher education. Finally, we found that teachers do not necessarily play a clear, central role in curriculum design. PMID- 19811149 TI - Social anxiety in medical students: Implications for communication skills teaching. AB - BACKGROUND: Social anxiety manifests as a fear of social situations, including being observed by others (Bruce & Saeed 1999). Communication skills workshops frequently involve student performance being observed by others, therefore social anxiety may impact upon attitudes to this style of teaching. AIMS: To determine the levels of social anxiety amongst medical undergraduates and investigate whether this influenced attitudes towards communication skills teaching. METHODS: 247 medical students (three year groups, 60% female) from the University of St Andrews completed a questionnaire survey measuring levels of social anxiety and attitudes to communication skills teaching (Mattick & Clarke 1998; Rees et al. 2002 ). RESULTS: Average social anxiety scores in the students were lower (t tests, P < 0.001) than other groups measured by Mattick & Clarke (1998). A minority (8%) of medical undergraduates however experienced high levels of social anxiety. High social anxiety scores correlated with negative attitudes to communication skills teaching, especially among female students (r = 0.359, P < 0.0001). CONCLUSIONS: Social anxiety contributes to a negative attitude towards communication skills teaching and may impact on participation in group workshops. This information could influence the methods tutors use for the provision of feedback in such workshops. PMID- 19811150 TI - Being able to be a Don Quixote! A different way of teaching clinical skills. PMID- 19811151 TI - The use of popular movies during lectures to aid the teaching and learning of undergraduate pharmacology. AB - BACKGROUND: The role of the lecturer has changed to one where they must engage and motivate students to learn the subject material. AIMS: To investigate whether the use of short movie references to pharmacology during lectures could stimulate learning in undergraduate students. METHODS: One- to two-min film clips from popular movies containing a reference to the subject being covered were incorporated into Powerpoint presentations and shown at different times during pharmacology lectures. At the end of the lecture series, a student survey was conducted to assess the impact of the movies on student motivation, engagement and learning. RESULTS: Three positive effects were noted. First, students related theory to (simulated) practice by recognising that what they had learnt was actually being used. Second, students were motivated to attend lectures to see what clip would be used. Third, the clips provided a sectioning break, which helped to maintain the engagement of students throughout the lecture as well as the organisation of the lecture by the lecturer. CONCLUSIONS: The use of short popular movie references was a novel way to motivate and maintain the interest of large classes of undergraduate students throughout lectures. PMID- 19811152 TI - Reliability of multi-station interviews in selection of junior doctors for specialty training. AB - BACKGROUND: Interviews for junior doctor training posts in the UK are considered high stakes assessments following the introduction of Modernizing Medical Careers. AIMS: To examine the reliability of paediatric recruitment in our Deanery for specialty training (ST) in 2008. METHODS: A total of 225 candidates were interviewed for 88 posts. There were three interview stations (presentation, structured interview and communication) each lasting for 10 min. Two interviewers independently marked candidates at each station on a 5 point Likert scale. Interview scores were analysed to evaluate reliability. RESULTS: Reliability for ST1 interviews was excellent, reasonable for ST2 and ST3 and poor for ST4 (Cronbach's alpha of 0.908, 0.749, 0.759 and 0.648). Generalizability coefficient was good for ST1 interviews (>0.8) but for ST2-ST4 interviews was below acceptable (<0.8). Decision (D) study calculations showed up to 26 stations that were required to achieve minimum reliability. CONCLUSIONS: This three station interview process was not reliable enough for a high stakes assessment. An increased number of stations are required to improve reliability. Further work is required to clarify the optimum number of stations to achieve minimum reliability and feasibility. PMID- 19811153 TI - Improving clinical examination, diagnosis and management skills in junior doctors managing hand injuries in the emergency department. PMID- 19811154 TI - Psychometric analyses and internal consistency of the PHEEM questionnaire to measure the clinical learning environment in the clerkship of a Medical School in Chile. AB - BACKGROUND AND AIMS: The Spanish version of the Postgraduate Hospital Educational Environment Measure (PHEEM) was evaluated in this study to determine its psychometric properties, validity and internal consistency to measure the clinical learning environment in the hospital setting of Pontificia Universidad Catolica de Chile Medical School's Internship. METHODS: The 40-item PHEEM questionnaire was translated from English to Spanish and retranslated to English. Content validity was tested by a focus group and minor differences in meaning were adjusted. The PHEEM was administered to clerks in years 6 and 7. Construct validity was carried out using exploratory factor analysis followed by a Varimax rotation. Internal consistency was measured using Cronbach's alpha. RESULTS: A total of 125 out of 220 students responded to the PHEEM. The overall response rate was 56.8% and compliances with each item ranged from 99.2% to 100%. Analyses indicate that five factors instrument accounting for 58% of the variance and internal consistency of the 40-item questionnaire is 0.955 (Cronbach's alpha). The 40-item questionnaire had a mean score of 98.21 +/- 21.2 (maximum score of 160). CONCLUSIONS: The Spanish version of PHEEM is a multidimensional, valid and highly reliable instrument measuring the educational environment among undergraduate medical students working in hospital-based clerkships. PMID- 19811155 TI - Initiation of a pediatric mock code program at a children's hospital. AB - BACKGROUND: Pediatric cardiopulmonary arrests are rare. Mock codes were instituted to bridge the gap between opportunity and reality. AIM: The goal was to improve medical caregivers' skills in pediatric resuscitation. METHODS: All pediatric and internal medicine/pediatric (med/peds) residents were anonymously surveyed pre- and post-intervention about confidence level about codes and code skills. Twenty mock codes were conducted during the 1 year intervention period. Statistical comparisons were made between each resident pre- and post-survey, graduating third-year residents (PGY3s) prior to intervention versus PGY3s with mock codes and pediatric versus med/peds residents. RESULTS: All residents significantly improved in their perception of overall skill level during the study (p < 0.0001). PGY3s were significantly more confident in their skills than PGY2s or PGY1s and PGY2s were significantly more confident than PGY1s both pre- and post-mock codes (p < 0.0001). Med/peds residents were significantly more confident in their skills than pediatric residents both pre- (p = 0.041) and post intervention (p = 0.016). The two skills with the lowest score post-intervention were the ability to place an interosseous line and the ability to manage cardiac dysrhythmias. CONCLUSIONS: Pediatric mock codes can improve resident confidence and self-assessment of their resuscitation skills. Data from surveys such as this can be used to design future skill-based educational initiatives. PMID- 19811156 TI - Meaningful learning: students' perceptions of a new form of case seminar in pathology. AB - BACKGROUND: Student-centred activities have been developed in a pathology course for medical students. AIM: This study reports on students' perceptions of a new form of case seminar as a way to learn pathology. METHOD: The seminar was evaluated through open-ended questionnaires and the data was analysed with a qualitative content analysis approach. RESULTS: All students reported that the case seminar was a positive learning experience. Four aspects of importance for learning were identified: motivational, knowledge construction, contextual and collaborative aspects. The motivational aspects concerned an increase in interest and motivation to learn, while the knowledge construction aspects included enhancing memory formation and facilitation of understanding. The case seminar also seems to help the students relate the textbook knowledge to a real world context and future profession, which can be described as the contextual aspects of learning. According to the students in our study, the work in small groups resulted in positive collaborative aspects of learning. CONCLUSIONS: The new case seminar could be an effective teaching and learning activity. It can be used in a traditional course as a complement to lectures and does not require a major change in the course design. It is also well suited for integrated curricula. PMID- 19811157 TI - Medical students' attitudes towards science and involvement in research activities: a comparative study with students from a reformed and a traditional curriculum. AB - BACKGROUND AND AIMS: Little is known about medical students' attitudes towards science and scientific methodology. We aimed to evaluate these attitudes and students' involvement in research activities. METHODS: Cross-sectional study comparing fifth-year medical students from the reformed and the traditional curriculum of the Charite University Medical Centre Berlin, Germany. Students filled out a standardised questionnaire containing three domains: research project for a dissertation; self-reported behaviour, knowledge/attitudes towards evidence-based medicine (EBM) and scientific methodology; and attitudes towards science. RESULTS: Two-thirds of the students had already started research for their dissertation and 70% agreed that reading articles and conducting research was challenging. Reformed curriculum students showed a higher involvement in scientific activities and felt more secure about their own scientific competencies. The odds for involvement in different research activities were significantly higher (odds ratios from 1.8 to 2.4) for students who agreed that 'science enables medical progress' or who felt 'secure in understanding medical articles and statistics' compared with students without these attitudes. CONCLUSION: The general attitude towards science and scientific methodology was positive among students from both the traditional and the reformed medical curriculum. Specific attitudes predicted involvement in research activities; however, they should be examined in other settings and student populations. PMID- 19811158 TI - Evaluation of a national process of reforming curricula in postgraduate medical education. AB - CONTEXT: A national reform of the postgraduate medical education in Denmark introduced (1) Outcome-based education, (2) The CanMEDS framework of competence related to seven roles of the doctor, and (3) In-training assessment. OBJECTIVES: The purpose of the study was to evaluate the process of developing new curricula for 38 specialist training programmes. The research question was: which conditions promote and which conditions impede the process? METHODS: Evaluation of the process was conducted among 76 contact-persons, who were chairing the curriculum development process within the specialties. Quantitative and qualitative data from a questionnaire survey and telephone interviews were triangulated for data analysis. RESULTS: The response rate of the questionnaire survey was 83% (63/76). Twenty-six telephone interviews were conducted. Identified promoting factors included positive attitude and motivation in faculty and support from written guidelines and seminars. Identified impeding factors included insufficient pedagogical support, poor introduction to the task, changing and inconsistent information from authorities, replacement of advisors, and stressful deadlines. CONCLUSIONS: This study identified promoting and impeding factors in a national postgraduate curriculum development process. Surprisingly the study indicates that pedagogical support provided throughout a process in some aspects might not be useful. General suggestions regarding curriculum reform processes are formulated. PMID- 19811159 TI - General physicians graduated from a PBL undergraduate medical curriculum: how well do they perform as PBL tutors? AB - BACKGROUND: A study was conducted on the effectiveness of general physicians recently graduated from a medical school with Problem-Based Learning (PBL) curriculum as PBL tutors to expand the school's tutor pool. AIMS: This study aims to investigate these non-staff tutors' effectiveness in terms of student satisfaction and learning outcomes. METHOD: An experimental study was conducted of 12 PBL groups of second-year medical students (n = 40). Four PBL groups were led by non-staff tutors; the other eight groups were led by staff tutors during the two PBL units. Tutor evaluation and student satisfaction questionnaires were administered and student performance scores were analysed to compare between groups led by staff tutors and non-staff tutors. RESULTS: The students' overall satisfaction with the non-staff tutors on a five-point Likert-scale was high (M = 4.5 +/-.638). Additionally, the student scores on written tests were comparable between groups. Yet, in one unit, the groups led by staff tutors received significantly higher scores on the group evaluation than those led by non-staff tutors. CONCLUSIONS: The results of this study show that the non-staff tutors performed as effectively as the staff tutors did with regard to student achievement in written exams. Still, the findings of this study suggest that different tutor backgrounds and experiences might affect student performance beyond the written exam scores. PMID- 19811160 TI - Medical students' attitudes towards disability and support for disability in medicine. AB - BACKGROUND: The number of medical students disclosing a disability is lower than the number of disabled doctors. AIMS: This study aimed to examine rates and types of disability in medical students, whether students disclosed this disability and their support needs, and, if not, possible reasons for non-disclosure. METHODS: This was a cross-sectional questionnaire study of all medical students registered at the University of Aberdeen in October 2007 (n = 944). RESULTS: Six percent of respondents considered they had a disability on application to medical school, but only 4% had disclosed this on admission. After reading the legal definition of disability, the percentage of respondents considering themselves as having a disability increased to 13%. Disabilities included: specific learning difficulties; mental health issues; sensory impairment; chronic illness; and mobility problems. Challenges to their studies due to disability were reported by nearly half of the disabled respondents but two-thirds had not sought support. Twelve percent reported experiencing discrimination due to their disability. Broader attitudes to disability indicated that many (75%) respondents believed some disabilities would prevent the study of medicine. CONCLUSIONS: Non disclosure of disability in medical students may be due to several factors including narrow definitions of disability and negative attitudes towards disability from the wider student body. PMID- 19811161 TI - What costs complexity and what price simplicity? PMID- 19811162 TI - The use of simulated patients in medical education: AMEE Guide No 42. AB - Medical training has traditionally depended on patient contact. However, changes in healthcare delivery coupled with concerns about lack of objectivity or standardization of clinical examinations lead to the introduction of the 'simulated patient' (SP). SPs are now used widely for teaching and assessment purposes. SPs are usually, but not necessarily, lay people who are trained to portray a patient with a specific condition in a realistic way, sometimes in a standardized way (where they give a consistent presentation which does not vary from student to student). SPs can be used for teaching and assessment of consultation and clinical/physical examination skills, in simulated teaching environments or in situ. All SPs play roles but SPs have also been used successfully to give feedback and evaluate student performance. Clearly, given this potential level of involvement in medical training, it is critical to recruit, train and use SPs appropriately. We have provided a detailed overview on how to do so, for both teaching and assessment purposes. The contents include: how to monitor and assess SP performance, both in terms of validity and reliability, and in terms of the impact on the SP; and an overview of the methods, staff costs and routine expenses required for recruiting, administrating and training an SP bank, and finally, we provide some intercultural comparisons, a 'snapshot' of the use of SPs in medical education across Europe and Asia, and briefly discuss some of the areas of SP use which require further research. PMID- 19811163 TI - A ten-year review of the literature on the use of standardized patients in teaching and learning: 1996-2005. AB - BACKGROUND: Although there is a growing body of literature on the educational use of standardized patients (SP) in teaching and learning, there have been no reviews on their value. OBJECTIVE: To determine whether the educational use of SPs has an effect on the knowledge, skills, and behaviour of learners in the health professions. METHODS: English-language articles covering the period 1996 2005 were reviewed to address the issue of to what extent has the use of SPs affected the knowledge, skills and performance of learners. Out of 797 abstracts, 69 articles, which met the review criteria, were selected. An adaptation of Kirkpatrick's model was used to classify and analyse the articles. RESULTS: Most of the learners were students in medicine and nursing. SPs were used mostly to teach communication skills and clinical skills. The study designs were case control (29%), pre-test/post-test (24.6%), post-test only (26.1%) and qualitative studies (20.3%). METHODOLOGICAL ISSUES: Most of the studies had weak research designs. More rigorous designs with control or comparison groups should be used in future research. CONCLUSIONS: Most studies reported that the educational use of SPs was valuable. More rigorous studies would support the evidence-based use of SPs in teaching and learning. PMID- 19811164 TI - Medical education in Libya: the challenges. AB - The history of medical education in Libya spans over a period of 40 years. Medical schools had a good and promising start in the 1970s. The graduates of the first few classes had a good impact on the health services in Libya. However, the medical schools did not embrace the immense changes that medical education experienced over the last two decades. This article aims to give a background on the medical education in Libya and explore the challenges facing it, which may help in gaining the initial momentum that seems to have been lost. PMID- 19811165 TI - Questionnaire survey for challenging cases of medical professionalism in Japan. AB - BACKGROUND: Little information is available on evaluation of medical professionalism among Japanese physicians and on its education in Japanese medical schools. AIMS: To assess professionalism and its education in Japan. METHOD: We analysed the responses to challenges to professionalism for Japanese residents and physicians, using the Barry Questionnaire, and to survey the extent of education related to professionalism during medical school curricula. The survey was conducted at 14 teaching hospitals in Kyushu and Okinawa, using existing hospital conferences. RESULTS: We collected data from 175 participants (60 residents and 115 faculty physicians). The most challenging was the sexual harassment scenario, in which 51.4% provided the best or 2nd best answers, followed by the honesty scenario with 69.7% and the confidentiality scenario with 76.0%. Participants were more likely to provide the best or 2nd best responses to the scenarios involving physician impairment (87.4%), conflict of interest (81.1%), and acceptance of gifts (78.3%). Five (3%) participants reported learning experiences about professionalism during the curricula and the median hours for its course work were two hours. Only one resident reported that she was satisfied with these educational sessions. CONCLUSIONS: Many Japanese physicians were unable to provide an acceptable response to challenges to professionalism in several issues and few had received education in professionalism during school curricula. Greater teaching of professionalism is needed in medical education in Japan. PMID- 19811166 TI - Randomized controlled trial comparing lecture versus self studying by an online tool. AB - OBJECTIVES: Since 2001, an e-learning tool has been developed for dentistry. To compare online self study with traditional lectures, a module about instrumental occlusal analysis was offered by oral lecture and by a unit in the virtual library of the e-learning tool. METHODS: 85 pre-clinical dental students were randomly divided into two groups: the computer assisted learning (CAL) group (n = 48) and the lecture group (n = 37). A pre-test was made to assess the students' basic knowledge. The first post-test and the scoring of the teaching methods were performed immediately after the lecture or the self studying by the online tool and a second post-test six weeks later. RESULTS: The oral lecture got better educational and enjoyment values. The students prefer CAL in addition to traditional lectures. The results of the pre-tests were not different between the two groups (p = 0.706). The lecture group significantly improved their scores in the first post-test (p = 0.011), but the scores of the second post-test did not differ significantly (p = 0.157). CONCLUSIONS: In the short term, knowledge acquisition seems to be better in oral lectures but in the long term there is no difference in knowledge retention between the two learning scenarios. PMID- 19811167 TI - Improving ethics education during residency training. AB - BACKGROUND: Trainees struggle with the evaluation and management of inpatient clinical ethical dilemmas. AIM: To meet their needs for both conceptual clarification and practical management, we designed a program to teach medical residents a systematic approach to resolving clinical ethical dilemmas. METHODS: We instituted monthly resident ethics educational case conferences to clarify residents' understanding of key concepts of medical ethics and to teach an 8-step systematic approach to resolving ethical dilemmas. We surveyed learners on the appropriateness, immediate utility, and potential for future usefulness of the approach. RESULTS: The vast majority of residents found the approach to be applicable and helpful with clinical decisions and interactions with patients and their family members. CONCLUSIONS: Teaching residents to use a systematic approach in understanding and resolving ethical dilemmas can facilitate their management of the ethical dilemmas that arise in clinical practice. Providing trainees with a concise structure for the thought process involved gives them confidence in their ability to address the issues directly and to act for reasons that are explicit, transparent, and reflect medical professionalism. PMID- 19811168 TI - Resources medical students use to derive a differential diagnosis. AB - BACKGROUND: Deriving an appropriate differential diagnosis is a key clinical competency, but there is little data available on how medical students learn this skill. Software resources designed to complement clinical reasoning might be asset in helping them in this task. AIMS: The goals of this study were to identify the resources third year medical students use to solve a challenging diagnostic case, and specifically to evaluate the usefulness of Isabel, a second generation electronic diagnosis support system. METHODS: Third year medical students (n = 117) were presented a challenging case and asked to identify and prioritize their top 3 diagnoses, report the time devoted to the exercise, and list the resources they used and their relative usefulness. Students were randomized to receive (or not) free access, instruction, and encouragement to use to a web-based decision support system (Isabel). RESULTS: Students who identified the correct diagnosis as their first choice spent significantly more time on the case than did the other students (3.75 +/- 0.28 hours vs 2.88 +/- 0.15 hours, p < 0.05). Students used electronic resources extensively, in particular Google. Students who self-reported use of Isabel had greater success identifying the correct diagnosis (24/33 = 73% for users vs 45/84 = 53% for non-users) a difference of borderline statistical significance. CONCLUSIONS: These findings indicate that medical trainees use a wide range of electronic decision support products to solve challenging cases. Medical education needs to adapt to this reality, and address the need to teach future clinicians how to use these tools to advantage. PMID- 19811169 TI - Talking with patients and peers: medical students' difficulties with learning communication skills. AB - BACKGROUND: Patient-centered communication skills, such as an empathic attitude towards patients and a holistic perspective on health, are difficult to acquire. Designing effective courses requires better understanding of the difficulties that students perceive with learning to talk with patients AIMS: The study aimed at exploring students' common difficulties with learning patient-centered communication skills. METHODS: Group discussions about student-patient interviews were videotaped and analyzed with regard to issues that students perceived as difficult and to their reflections about these difficulties. RESULTS: The students reported feeling intrusive as they explored the patient's psychosocial situation. They avoided being empathic and felt insecure about coping adequately with emotionally loaded topics. Their difficulties were mainly due to insufficient understanding of the functional relations between psychosocial issues and health conditions. Moreover, students were insecure concerning the function of affective feedback in the diagnostic process. However, the group discussions generated a language for analyzing and structuring interviews that helped develop the students' professional identities. CONCLUSIONS: Students experienced moral qualms about applying major aspects of patient-centered interviewing. Instruction in communication skills should aim at filling the students' knowledge gaps and fostering their awareness and expression of emotional perceptions. Long-term relationships with patients could help develop patient-centered communication. PMID- 19811170 TI - Comparing staff and student perceptions of the student experience at a new medical school. AB - AIM: The aim was to compare staff and student perceptions of student experience with the UEA MB/BS educational environment. METHODS: All MB/BS students were asked to complete a standard course evaluation at the end of the 2005/06 academic year, which included the Dundee Ready Education Environment Measure (DREEM). MB/BS teaching staff were asked to complete a revised version of the DREEM. RESULTS: Data from 403 students and 73 teaching staff were compared. The overall DREEM was 144/200 for staff and 141/200 for students. Initial analysis suggested that staff believed that students' were experiencing more positive learning and teaching environments than students actually were, and they viewed the students' social experience as more negative than it actually was. However, staff were unable to comment on number of aspects of the educational environment and scored these as 'unsure'. This distorted the mean values for the subscales. When this was adjudged for, the differences disappeared. CONCLUSIONS: Staff did not believe that students were experiencing a perfect educational environment; in most cases their views were closely aligned. But staff 's unfamiliarity with aspects of the educational environment suggests a continuing need to provide feedback to staff about students' actual experiences, to enable provision of a better environment for students. PMID- 19811171 TI - Freeware eLearning Flash-ECG for learning electrocardiography. AB - BACKGROUND: Electrocardiographic (ECG) analysis can be taught in eLearning programmes with suitable software that permits the effective use of basic tools such as a ruler and a magnifier, required for measurements. AIMS: The Flash-ECG (Research & Development Unit for Medical Education, University of Helsinki, Finland) was developed to enable teachers and students to use scanned and archived ECGs on computer screens and classroom projectors. RESULTS: The software requires only a standard web browser with a Flash plug-in and can be integrated with learning environments (Blackboard/WebCT, Moodle). The Flash-ECG is freeware and is available to medical teachers worldwide. PMID- 19811172 TI - DREEM, PHEEM, ATEEM and STEEM in Japanese. PMID- 19811173 TI - A game for teaching antimicrobial mechanisms of action. AB - BACKGROUND: Alternative teaching tools have proved to enhance students' interest and knowledge skills. AIM: To integrate basic Bacteriology with mechanisms of action of antimicrobial agents. METHODS: The board has 121 squares, including squares with question marks and antimicrobial agents. Each student receives a card with a clinical case, identification of the bacterium and its resistance to antimicrobials. The student rolls a dice and moves the corresponding number of squares. The game depends on the dice values rolled, the bacterial resistance profile, and the questions the student has to answer each time he/she lands on a question mark. Previously, the students were given a lecture about the subject. On the day of the game, students answered a pre-test and a post-test. The paired t-test was used for the statistical analysis. RESULTS: The game was applied to 78 students of the Medicine and Pharmacy undergraduate courses of the Universidade Federal do Rio Grande do Sul, Brazil. There was an increase in the number of right answers and a decrease in the number of unknown answers. There were no significant differences between the courses. CONCLUSION: The game could be applied to other undergraduate courses in the field of Health Sciences. PMID- 19811174 TI - Is learning anatomy facilitated by computer-aided learning? A review of the literature. AB - BACKGROUND: There is ongoing debate concerning the best way to teach anatomy. Computer-assisted learning (CAL) is one option for teaching anatomy and these resources are increasingly available. AIMS: To assess the use of such resources in undergraduate medical student anatomy tuition. METHOD: Literature review. RESULTS: Eight quantitative studies were found and these tended to report favourably. Though these educational packages can show improvement in knowledge, the studies tended to cover small areas of anatomy or were assessed in short courses. There were also several assessments of learner's attitudes to CAL which tended to report favourably in terms of educational satisfaction and enjoyment. CONCLUSIONS: There is insufficient evidence to show that these resources have a true place for replacing traditional methods in teaching anatomy. Further research should be conducted to determine how to use these resources in conjunction with current teaching methods or how their use can be integrated into the current anatomy curriculum. PMID- 19811175 TI - Medical education in Korea: the e-learning consortium. AB - This article reports the latest development in e-learning in Korean medical education. The Korean Consortium for e-Learning in Medical Education was formed for collaboration in providing quality online learning resources for medical schools around the nation. This e-learning strategy is aimed at improving the quality of medical education at the national level by providing students with equal access to quality learning resources and fostering students' self-directed learning and, in doing so, enhancing the effectiveness and efficiency of developing online learning resources by sharing necessary resources among the medical schools. The consortium also plans to share e-learning content with medical schools in other countries by engaging more medical schools in the consortium and also by sharing e-learning content developed by other institutions or consortiums. The consortium is also dedicated to the research and development of effective online learning strategies for medical education, including interactive virtual patient cases and other innovative pedagogies using Web 2.0 technologies. PMID- 19811176 TI - A study of physiotherapy students' and clinical educators' perceptions of learning and teaching. AB - BACKGROUND: Clinical education is a key component to learning in the health professions. AIMS: This qualitative study investigated how physiotherapy students and clinical educators perceived their respective roles in learning and teaching clinical skills during students' first clinical placements. METHODS: Separate physiotherapy student and educator focus groups were conducted in two major teaching hospitals in Melbourne, Australia during students' first clinical placements. RESULTS: The key difference between students' and educators' perspectives of their role was their description of how to build knowledge within clinical placement settings. Clinical educators' focused on steps involved in their teaching tasks, rather than ways to facilitate learning. Their conception of teaching was to impart structured knowledge to students in response to knowledge deficits. Students also identified knowledge gaps but they proposed alternative practical ways to build their knowledge. Their conception of learning was to move from an initial static identification of knowledge deficits, to a more dynamic search for methods and people that might build their knowledge and understanding. CONCLUSIONS: The findings of this research were used to develop a set of teaching and learning statements and strategies that are grounded in the perspectives and experiences of students and educators in the clinical education setting. PMID- 19811177 TI - What criteria do faculty use when rating students as potential house officers? AB - Third-year medical students' grades are a combination of faculty observations, National Board of Medical Examiners examinations, and other departmental specific course requirements. Faculty evaluations include assessment of students' clinical skills, and a global rating for potential as house officer. We wished to better understand the 'potential as house officer' and to understand if these competencies were shared across the third year or unique to a given discipline. We then examined the relationship between house officer potential and performance on traditional measures of success. We analyzed the narrative comments from faculty evaluations of third-year students who faculty rated as 'Outstanding' in the house officer potential category. The low correlations found between house officer potential and traditional measures of academic success indicate that items beyond the stated learning objectives are influencing faculty evaluation of clinical students. Our data suggest that the awarding of 'potential for house officer' reflects the student's ability to work as part of a health care team. Although there appear to be common elements among the house officer comments, we also observed discipline specific differences. Given the importance placed on house officer potential, more conversation is needed to develop a common language across the third-year courses. PMID- 19811178 TI - Factors that influence first year medical students' choice of student selected component. AB - BACKGROUND: Undergraduate medical training should ensure students have choice and autonomy in the learning process, including the student selected components (SSCs) which should comprise up to about a third of the curriculum. Students' choices of SSC will influence the knowledge, skills and attitudes they acquire. AIM: To investigate how motivations and personality in first year medical students influence their choice of SSCs. METHOD: A questionnaire regarding motivations for SSC choice and the NEO-FFI personality measure was administered to all first year students at a London medical school. Relationships with type of SSC were examined. RESULTS: A total of 82% (268/329) students responded. Six motivational factors arose from a principle components analysis of the questionnaire: future achievements, prior information, internal motivation, personal recommendation, convenience and certainty. Students with different motivational factors chose different SSCs, and had different personality traits. Weak but significant correlations were found between personality traits and motivational factors, but not between personality and SSC choice, or sex and SSC choice. CONCLUSIONS: This offers insight into medical student choices of SSC and is the first step towards ensuring appropriate provision of modules that students wish to study to enable them to meet the demands of the medical profession. PMID- 19811179 TI - Effects of "minimally invasive curricular surgery" - a pilot intervention study to improve the quality of bedside teaching in medical education. AB - BACKGROUND: Bedside teaching is an important element of undergraduate medical education. However, the impact of curricular course structure on student outcome needs to be determined. AIMS: This study assessed changes in fourth-year medical students' evaluations of clinical teaching sessions before and after the introduction of a new course format. METHOD: The curricular structure of bedside teaching sessions in cardiology was modified without changing the amount of teaching time. Clinical teachers were instructed about the new teaching format and learning objectives. The new format implemented for adult but not paediatric cardiology sessions was piloted with 143 students in winter 2007/08. By computing effect sizes, evaluation results were compared to data obtained from 185 students before the intervention. RESULTS: Significant rating increases were observed for adult cardiology teaching sessions (Cohen's d = 0.66) but not paediatric cardiology sessions (d = 0.22). In addition to improving the structure and organization of the course, the intervention significantly impacted on students' perceptions of their learning outcome regarding practical skills (d = 0.69). CONCLUSIONS: Minimal curricular changes combined with basic faculty development measures significantly increase students' perception of learning outcome. Curricular structure needs to be considered when planning bedside teaching sessions in medical undergraduate training. PMID- 19811180 TI - Web-based collaborative training of clinical reasoning: a randomized trial. AB - BACKGROUND: Clinical reasoning skills are essential for medical practice. Problem based collaborative learning via the internet might prove useful in imparting these skills. AIM: This randomized study assessed whether web-based learning (WBL) is superior to face-to-face problem-based learning (PBL) in the setting of a 6-week cardio-respiratory course. METHODS: During winter term 2007/08, all 148 fourth-year medical students enrolled in the 6-week course consented to be randomized in small groups to diagnose a patient complaining of dyspnoea either using a virtual collaborative online module or a traditional PBL session. Clinical reasoning skills were assessed by means of a key feature examination at the end of the course. RESULTS: No significant difference between the mean scores of both study groups was detected (p = 0.843). In virtual learning groups, costs for diagnostic tests were significantly correlated to the number of contributions to online group discussions (r = 0.881; p = 0.002). Evaluation data favored traditional PBL sessions over virtual collaborative learning. CONCLUSION: While virtual collaborative learning was as effective as traditional PBL regarding the acquisition of clinical reasoning skills, it was less well accepted than traditional PBL. Future research needs to determine the ideal format and time point for computer-assisted learning in medical education. PMID- 19811181 TI - Effects of a supplementary final year curriculum on students' clinical reasoning skills as assessed by key-feature examination. AB - BACKGROUND: The final year of medical education is considered crucial in making students 'fit for purpose'. Studies have shown that many students leave medical school without having experienced sufficient preparation for their upcoming professional life. AIM: The aim of this study was to examine the effectiveness of a supplementary internal medicine final year curriculum on clinical reasoning skills. METHOD: Final year internal medicine students from two universities participated in the study which was based on a static-group design. The experimental group (n = 49) took part in a final year student curriculum with interactive case-based seminars and skills training sessions. The comparison group (n = 25) did not receive any additional training beyond working on the ward. Clinical reasoning skills were assessed using a key-feature pre-post test. RESULTS: Prior to their clinical rotation, the two groups did not differ in the key-feature examination (p < 0.924). The experimental group performed significantly better than the comparison group (p < 0.028) in the post intervention key-feature examination. CONCLUSIONS: Supplementary interactive case based seminars and skills training sessions are effective and significantly improve the clinical reasoning skills of final year students in internal medicine. Further study is warranted and should look to examine the effectiveness of a final year student curriculum on other performance measures. PMID- 19811182 TI - Basic sciences in medical education: why? How? When? Where? PMID- 19811183 TI - Portfolios for assessment and learning: AMEE Guide no. 45. AB - In 1990, Miller wrote that no tools were available for assessment of what a learner does when functioning independently at the clinical workplace (Miller 1990 ). Since then portfolios have filled this gap and found their way into medical education, not only as tools for assessment of performance in the workplace, but also as tools to stimulate learning from experience. We give an overview of the content and structure of various types of portfolios, describe the potential of electronic portfolios, present techniques and strategies for using portfolios as tools for stimulating learning and for assessment, and discuss factors that influence the success of the introduction. We conclude that portfolios have a lot of potential but that their introduction also often leads to disappointment, because they require a new perspective on education from mentors and learners and a significant investment of time and energy. PMID- 19811184 TI - Training of physicians for the twenty-first century: role of the basic sciences. AB - BACKGROUND: Rapid changes in the healthcare environment and public dissatisfaction with the cost and quality of medical care have prompted a critical analysis of how physicians are trained in the United States. Accrediting agencies have catalyzed a transformation from a process based to a competency based curriculum, both at the undergraduate and the graduate levels. AIM: The objective of this overview is to determine how these changes are likely to alter the role of basic science in medical education. METHODS: Policy statements related to basic science education from the National Board of Medical Examiners (NBME), the Accreditation Council for Graduate Medical Education (ACGME), American Board of Medical Specialties (ABMS), and the Federation of State Medical Boards (FSMB) were reviewed and assessed for common themes. RESULTS: Three primary roles for the basic sciences in medical education are proposed: (1) basic science to support the development of clinical reasoning skills; (2) basic science to support a critical analysis of medical and surgical interventions ("evidence-based medicine"); and (3) basic and translational science to support analysis of processes to improve healthcare ("science of healthcare delivery"). CONCLUSION: With these roles in mind, several methods to incorporate basic sciences into the curriculum are suggested. PMID- 19811185 TI - Teaching basic science to optimize transfer. AB - BACKGROUND: Basic science teachers share the concern that much of what they teach is soon forgotten. Although some evidence suggests that relatively little basic science is forgotten, it may not appear so, as students commonly have difficulty using these concepts to solve or explain clinical problems: This phenomenon, using a concept learned in one context to solve a problem in a different context, is known to cognitive psychologists as transfer. The psychology literature shows that transfer is difficult; typically, even though students may know a concept, fewer than 30% will be able to use it to solve new problems. However a number of strategies to improve transfer can be adopted at the time of initial teaching of the concept, in the use of exemplars to illustrate the concept, and in practice with additional problems. AIM: In this article, we review the literature in psychology to identify practical strategies to improve transfer. METHODS: Critical review of psychology literature to identify factors that enhance or impede transfer. RESULTS: There are a number of strategies available to teachers to facilitate transfer. These include active problem-solving at the time of initial learning, imbedding the concept in a problem context, using everyday analogies, and critically, practice with multiple dissimilar problems. Further, mixed practice, where problems illustrating different concepts are mixed together, and distributed practice, spread out over time, can result in significant and large gains. CONCLUSION: Transfer is difficult, but specific teaching strategies can enhance this skill by factors of two or three. PMID- 19811186 TI - No content without context: integrating basic, clinical, and social sciences in a pre-clerkship curriculum. AB - BACKGROUND: The basic science curricula in medical schools ultimately succeed or fail at the bedside when students must draw on their pre-clerkship experiences as they learn to form nuanced clinical decisions. Given this expectation, learning context becomes as decisive as content in determining students' recall and application. AIMS: Using the pre-clerkship medical curriculum at the University of California, Los Angeles, as an example, we illustrate how traditional biomedical sciences can be integrated with clinical sciences in a comprehensive foundational curriculum following curricular design features and teaching methods based on learning principles from cognitive psychology and education. METHODS: Multiple planning teams of faculty and students collaborated in the design of the Human Biology and Disease (HB&D) curriculum. Broad participation, careful selection of course chairs, the assistance of educational consultants, ongoing oversight structures, and faculty development were used to develop and sustain the curriculum. RESULTS: The resulting HB&D curriculum features an interdisciplinary spiral block structure including interactive lecture formats, integrative formative and summative examinations, self- and peer-taught laboratories, and problem-based learning with innovative variations. CONCLUSION: Our fully integrated, spiral, pre-clerkship curriculum built on repeating interdisciplinary blocks and longitudinal threads has yielded encouraging results as well as some specific innovations that other schools or individual teachers may find valuable to adapt for use in their own settings. PMID- 19811187 TI - Key steps for integrating a basic science throughout a medical school curriculum using an e-learning approach. AB - Basic sciences can be integrated into the medical school curriculum via e learning. The process of integrating a basic science in this manner resembles a curricular change. The change usually begins with an idea for using e-learning to teach a basic science and establishing the need for the innovation. In the planning phase, learning outcomes are formulated and a prototype of the program is developed based on the desired requirements. A realistic concept is formed after considering the limitations of the current institute. Next, a project team is assembled to develop the program and plan its integration. Incorporation of the e-learning program is facilitated by a well-developed and communicated integration plan. Various course coordinators are contacted to determine content of the e-learning program as well as establish assessment. Linking the e-learning program to existing course activities and thereby applying the basic science into the clinical context enhances the degree of integration. The success of the integration is demonstrated by a positive assessment of the program including favourable cost-benefit analysis and improved student performance. Lastly, when the program becomes institutionalised, continuously updating content and technology (when appropriate), and evaluating the integration contribute to the prolonged survival of the e-learning program. PMID- 19811188 TI - Development of basic medical sciences in a new medical school with an integrated curriculum: the ANU experience. AB - BACKGROUND: The development of a basic medical science curriculum in a new medical school with a problem-based focus in Australia has been subject to a number of constraints. We describe the process and early evaluation. AIM: To describe the development of a basic medical science curriculum in an Australian medical school with a problem-based curriculum. METHODS: We describe the process we used for curriculum development and the benefits and constraints that arose from pre-existing strong biomedical science on the Australian National University (ANU) campus. We outline methods we used to inform our curriculum content and report on accreditation and early internal evaluation. RESULTS: Australian medical schools design their curriculum within a relatively restrictive framework put forward by a national accreditation system. The curriculum achieved accreditation from the external accrediting agency, but early student evaluation has been mixed. CONCLUSION: Although our internal faculty evaluation and external review by the accrediting agency has supported the view that this aspect of the curriculum has performed reasonably well, student feedback is mixed and further evaluation is needed and adjustments probably warranted. PMID- 19811189 TI - Case method teaching: an effective approach to integrate the basic and clinical sciences in the preclinical medical curriculum. AB - BACKGROUND: Recent efforts to identify the essential skills and competencies required for medical practice have resulted in an expansion of the educational outcomes for which medical schools are accountable. Teachers in the preclinical years, formerly focused on the transmission of biomedical principles and factual information, are now charged with presenting discipline-specific concepts with an emphasis on clinical relevance while advancing active learning, critical thinking, communication skills, and other professional competencies. Problem based learning has been widely introduced to support these educational goals but other, less resource-intensive, discussion methodologies have not been extensively explored. AIM: To examine the feasibility of case-method teaching (CMT) during the preclinical curricula to integrate basic science concepts in the management of clinical problems. METHODS: CMT sessions were conducted with students during the first- and second-year of hybrid curricula at two US medical schools. RESULTS: First- and second-year medical classes of 40-95 students prepared for and actively engaged in single session case discussions and were able to productively apply basic science principles in clinical problem-solving. CONCLUSION: CMT represents a feasible and resource-conservative pedagogical format to promote critical thinking and to integrate basic science principles during the preclinical curriculum. PMID- 19811190 TI - The use of simulation in medical education to enhance students' understanding of basic sciences. AB - Simulation! The deans talk about its use in medical education regularly. But how can it be used to teach the basic sciences to first- and second-year medical students? This article will help answer that question by providing information about various types of simulation activities being used in medical education and examples of their application to basic science education. The next step depends on your creativity. PMID- 19811191 TI - Teaching of medical pharmacology: the need to nurture the early development of desired attitudes for safe and rational drug prescribing. AB - Pharmacology, as a basic medical science discipline, provides the scientific basis of therapeutics, i.e. the scientific foundation for safe and rational prescribing of drugs. The public, lay media, and the medical profession have raised serious concerns over the high incidence of errors of drug prescribing which compromise patient safety, including death of some patients, attributed mainly to inadequate teaching of medical pharmacology and, consequently, to medical graduates lacking skills in safe and effective drug prescribing. There is also overwhelming evidence that the pervasive and prevalent doctor-drug industry relationships have a strong influence over the prescribing habits and drug education of doctors. The British Pharmacological Society and American Association of Medical Colleges have crafted some insightful guidelines, including the learning of desired attitudes, for designing a medical pharmacology curriculum aimed at enhancing patient safety. This article will critically review the major issues relating to errors of drug prescribing, including the need to nurture the early development of desired attitudes which foster safe and rational drug prescribing. A simple educational approach, using a task analysis of drug prescribing, is applied to identify desired attitudes which should be incorporated into a basic pharmacology course for medical students in the twenty first century. PMID- 19811192 TI - Restructuring a basic science course for core competencies: an example from anatomy teaching. AB - Medical schools revise their curricula in order to develop physicians best skilled to serve the public's needs. To ensure a smooth transition to residency programs, undergraduate medical education is often driven by the six core competencies endorsed by the Accreditation Council for Graduate Medical Education (ACGME): patient care, medical knowledge, practice-based learning, interpersonal skills, professionalism, and systems-based practice. Recent curricular redesign at Mayo Medical School provided an opportunity to restructure anatomy education and integrate radiology with first-year gross and developmental anatomy. The resulting 6-week (120-contact-hour) human structure block provides students with opportunities to learn gross anatomy through dissection, radiologic imaging, and embryologic correlation. We report more than 20 educational interventions from the human structure block that may serve as a model for incorporating the ACGME core competencies into basic science and early medical education. The block emphasizes clinically-oriented anatomy, invites self- and peer-evaluation, provides daily formative feedback through an audience response system, and employs team-based learning. The course includes didactic briefing sessions and roles for students as teachers, leaders, and collaborators. Third-year medical students serve as teaching assistants. With its clinical focus and competency based design, the human structure block connects basic science with best-practice clinical medicine. PMID- 19811193 TI - Intercultural competence in medical education - essential to acquire, difficult to assess. AB - Determining student acquisition of intercultural competence (awareness, skills, knowledge and attitudes) is necessary in medical courses. However, addressing students' learning needs and developing effective tools to measure development of intercultural competence is challenging. Where this is done inadequately, skills may be overlooked or simplistic, one dimensional notions of culture be reinforced. This article examines aspects of the OSCE station development process, raising questions about how and when to assess acquisition of IC in undergraduate medical courses. It cautions against development of assessment tools which may lack authenticity and require students to engage in interactions which are unnatural. It argues for skills consistent with IC to be viewed as part of, and not separate from, the broad spectrum of skills which are a feature of any sensitive and appropriate doctor-patient interaction. Finally it advocates careful consideration of the optimum time to assess students' capacity to demonstrate IC in their interactions with patients, peers and staff, suggesting that this should come in later rather than earlier years, following theoretical, experiential and reflective learning. PMID- 19811194 TI - Teaching chronic condition management to undergraduate medical students: utilising the patient-as-teacher approach. PMID- 19811195 TI - Education format and resource preferences among registrants of a pediatric focused CME website. AB - BACKGROUND: Despite dramatic increase in Internet-based CME activities, little is known about physician Internet CME preferences. AIMS: To identify the education format and resource preferences among registrants of a pediatric-focused CME website. METHODS: Preferences of physician registrants at PedsEducation.org between July 2000-November 2007 (n = 1388) were assessed via survey. A secondary analysis of respondent demographics vs. reported preferences was conducted. RESULTS: A total of 345 physicians participated (25% response rate). The majority (73%, n = 252) identified free CME as a highly important feature of an Internet CME resource; monthly case series was identified as the least important. Seventy five percent of respondents (n = 260) identified practice guideline updates as a highly useful practice resource; practice feedback was identified as the least useful. Respondents with < or =10 years practice experience were more likely to identify case-based CME as highly useful to their daily practice (p < 0.001); respondents who spend > or = 90% working time on patient care were more likely to identify Internet CME as a highly useful CME format (p < 0.001). CONCLUSION: Internet CME preferences of PedsEducation.org registrants differ from those typically associated with knowledge gains and behavior changes. Demographic characteristics may influence these preferences. PMID- 19811196 TI - New simulation-based airway management training program for junior physicians: Advanced Airway Life Support. AB - BACKGROUND: All junior physicians in Taiwan were enrolled into a 3-month post graduate year 1 (PGY1) course after Severe Acute Respiratory Syndrome (SARS) attack in 2003. AIMS: To develop and evaluate a new airway management training protocol by using an integrated course of lectures, technical workshops and medical simulations. METHODS: In each PGY1 course, the trainees participated in the Advanced Airway Life Support (AALS) program. After 2 h lecture, the trainees were divided into three groups for 4 h technical workshop, including 10 skill stations and medical simulation at the Clinical Skills Resources Center of the hospital at different times. Video-based debriefing and feedback were performed after each simulation. The same scenario was re-simulated after debriefing. Participants' performance was assessed by single global rating and a 5 key actions scoring. RESULTS: A total of 266 junior physicians have been trained with this AALS programs in 2 years. They learned the techniques of airway management, passed the performance checklist of technical workshop, and received higher scores during re-simulation regardless of scoring methods. CONCLUSIONS: The AALS training program can provide methodical and systematic training for junior residents to mature with specialized technical skills and higher-order cognitive skills, behaviors and leadership in airway management. PMID- 19811197 TI - Training the intern: The value of a pre-intern year in preparing students for practice. AB - AIMS: To evaluate the clinical and professional development that occurs during a New Zealand trainee intern year in preparation for the first house officer role. METHODS: A quantitative questionnaire was distributed to all trainee interns (year 6) and year 5 medical students in New Zealand at the end of the 2007 academic year. This survey assessed self-reported competency and performance across clinical, professional and role development domains. RESULTS: Response rate was 65% (457/702). Compared to year 5 students, trainee interns reported significantly greater competence and performance levels across all three domains. The greatest improvement occurred in the independent performance of procedural skills (trainee interns: 77%, year 5: 35%, p < 0.001) and clinical tasks (trainee interns: 94%, year 5: 56%, p < 0.001) and in the level of clinical responsibility taken (p < 0.001). At the end of the trainee intern year, 92% of students felt prepared to be a junior doctor, versus only 53% at the end of their 5th year (p < 0.0001). CONCLUSIONS: The trainee intern year is important in preparing graduates for the intern role. The year affords increased responsibility and practical experience, whilst retaining an educational focus, facilitating the move from competence towards performance. Preparedness for practice was substantially higher following the New Zealand trainee intern year than has been reported with other pre-intern placements. PMID- 19811198 TI - Medical informatics in the internal medicine clerkship: Results of a national survey. AB - BACKGROUND: Medical Informatics (MI) is increasingly a critical aspect of medical education and patient care. AIMS: This study assessed the status of MI training, perception of needs and barriers for the implementation of MI curricula and utilization of information technology (IT) in patient care and medical education. METHOD: The MI questionnaire was a part of the 2006 Clerkship Directors in Internal Medicine survey of 110 institutional members. Descriptive statistics were calculated using Statistical Package for the Social Sciences (SPSS), version 12 and all p-values are two-tailed. RESULTS: Eighty-three (75%) members responded. Out of this, 52, 32.5 and 12% report that students receive MI training for patient care activities during pre-clinical years, third-year internal medicine clerkship or intersession, respectively. House staff critiques (46.4%), patient billing (44.1%), radiographic imaging (40.8%), accessing clinical data (37.3%), and student evaluations (36.1%) were areas in which 35% of respondents use IT 'all the time.' Fifty-one percent of respondents rate the adequacy of training in MI as average. Cost, time and lack of trained faculty were primary barriers for the implementation. CONCLUSIONS: Significant variations exist in timing of MI curricula. IT is utilized more frequently for non-patient activities. Studies are needed to examine the needs, processes and outcomes of MI curricula. PMID- 19811199 TI - Measuring professionalism in residency training programs in Iran. AB - BACKGROUND AND AIM: To assess the Farsi (Persian) translated and modified version of the questionnaire of the American Board of Internal Medicine (ABIM) for measuring professionalism, and also, measuring the professional attitudes and behaviors associated with the medical residency training environment in Iran. METHODS: After a pilot study, a 17-item questionnaire was distributed to 282 medical residents of two major universities of Iran, from December 2006 through February 2007. RESULTS: Of the 282 distributed questionnaires, 259 (95.2%) were used in the analysis. Based on the analyses, two items were omitted and 15 items were retained for further analysis. The mean score was 106 (Standard Deviation (SD), 22.4) out of maximum 150, whereas the item mean was 6.12 (SD, 0.37) out of maximum 10. Corrected item-to-total correlations ranged from low to moderate. The internal reliability of the scale, based on Cronbach's alpha, meets Nunnally's minimal requirement. A factor analysis was performed, based on principal components and varimax rotation. The solution identified three factors (subscales) including excellence, honor/integrity and altruism/respect. Together these factors represented 58.8% of the common variance. CONCLUSION: This study showed the content validity and internal reliability of the Farsi version of the ABIM questionnaire. Therefore, it can be considered as an encouraging step toward developing a short, reliable and valid instrument for measuring professionalism in medical environments. PMID- 19811200 TI - The role of observational research in improving faculty lecturing skills: A qualitative study in an Italian dental school. AB - BACKGROUND: This pilot study is based on observational research of lecturing skills during the annual Oral Medicine course at the Milan Dentistry School. AIMS: Our goals were to explore how teachers exhibited desirable lecturing skills, to observe how their attitudes and lecturing skills affected students' attention and thereby learning, and to provide feedback. METHOD: We prepared a structured observational grid divided into four categories: explaining, questioning, visual aids, and lecturer attitude. The grid was filled in by a participant, nonactive researcher. RESULTS: Two main types of lecture were observed: "traditional" and "interactive". Both of these can result in a high level of attention among students. Among the categories, only "lecturer attitude" appeared to affect student attention. In particular, the skills of "speaking aloud" and "sustaining verbal communication with vocal inflection" appeared to have the greatest impact on lecturer attitude. The data were then presented blindly to the five lecturers, who were able to identify their own lesson. CONCLUSIONS: Our grid proved to be a valid instrument although it was very expensive. When integrated with other strategies for improving lecturing, such as student scoring, peer evaluation, and microteaching, observational research can be a cost-effective method to stimulate guided reflection and to improve the lecturing skills of faculty members. PMID- 19811201 TI - Attitudes to patient safety amongst medical students and tutors: Developing a reliable and valid measure. AB - BACKGROUND: Patient safety education is an increasingly important component of the medical school curricula. AIMS: This study reports on the development of a valid and reliable patient safety attitude measure targeted at medical students, which could be used to compare the effectiveness of different forms of patient safety education delivery. METHODS: The Attitudes to Patient Safety Questionnaire (APSQ) was developed as a 45-item measure of attitudes towards five patient safety themes. In Study 1, factor analysis conducted on the responses of 420 medical students and tutors, revealed nine interpretable factors. The revised 37 item APSQ-II was then administered to 301 students and their tutors at two further medical schools. RESULTS: Good stability of factor structure was revealed with reliability coefficients ranging from 0.64 to 0.82 for the nine factors. The questionnaire also demonstrated good criterion validity, being able to distinguish between tutors and students across a range of domains. CONCLUSIONS: This article reports on the first attempt to develop a valid and reliable measure of patient safety attitudes which can distinguish responses between different groups. The predictive validity of the measure is yet to be assessed. The APSQ could be used to measure patient safety attitudes in other healthcare contexts in addition to evaluating changes in undergraduate curricula. PMID- 19811202 TI - Clinical teaching improvement: The transportability of the Stanford Faculty Development Program. AB - BACKGROUND: The Stanford Faculty Development Center (SFDC) at Stanford University developed a teaching improvement course for medical teachers that has been widely disseminated using a train-the-trainer model. We were curious to see if cultural factors might influence the applicability and impact of the course when delivered to non-American participants by a facilitator from that culture. METHODS: A Swedish anaesthesiologist at Uppsala University Hospital, Sweden, was trained in October 2004 at Stanford University. From January 2005 to March 2007 he delivered five faculty development seminar series at Uppsala University Hospital to 40 physicians from different departments. Participants rated the usefulness of the seminar series and retrospective pre- and post-seminar ratings were used to assess effects on participants' teaching skills and behaviours. RESULTS: Participants rated the seminars as highly useful (M = 4.8, SD = 0.4). Participants' ratings of their teaching ability indicated significant increases across a variety of clinical and non-clinical teaching settings (p < 0.001), and positive changes in teaching behaviours were found for all seven educational categories assessed (p < 0.001). CONCLUSIONS: This faculty development model is highly transportable to medical teachers in Sweden, and capable of producing positive results, consistent with those found in the United States. PMID- 19811203 TI - Virtual patients come of age. AB - On-screen simulations of clinical settings have been used for educational purposes since the 1970s. Despite this, it is only now that these 'virtual patients' are increasingly forming a part of the medical education mainstream. Enabling factors for these changes include a requirement for more assured clinical encounters, changes in patient availability (in particular, in tertiary contexts), diminishing technical and cost barriers and ongoing changes in educational practices as a whole. This special edition of Medical Teacher presents a number of papers covering key factors in the development, use and evaluation of virtual patients in contemporary medical education practice. PMID- 19811204 TI - The use of reflection in medical education: AMEE Guide No. 44. AB - Reflection is a metacognitive process that creates a greater understanding of both the self and the situation so that future actions can be informed by this understanding. Self-regulated and lifelong learning have reflection as an essential aspect, and it is also required to develop both a therapeutic relationship and professional expertise. There are a variety of educational approaches in undergraduate, postgraduate and continuing medical education that can be used to facilitate reflection, from text based reflective journals and critical incident reports to the creative use of digital media and storytelling. The choice of approach varies with the intended outcomes, but it should also be determined by the user since everyone has a preferred style. Guided reflection, with supportive challenge from a mentor or facilitator, is important so that underlying assumptions can be challenged and new perspectives considered. Feedback also has an important role to enhance reflection. There is little research evidence to suggest that reflection improves quality of care but the process of care can be enhanced. PMID- 19811205 TI - Medical education in Ireland. AB - Medical education in Ireland has undergone significant recent reform. The entire continuum from undergraduate, postgraduate and independent practitioner has been included in this reform process. This article looks at these changes within the Irish medical education system and what impact these changes are likely to have on the future training and practice of medicine within the country. PMID- 19811206 TI - The role of intellectual property in creating, sharing and repurposing virtual patients. AB - Medical schools are integrating more technology into the training of health care practitioners. Electronic Virtual Patients (VPs) provide interactive simulations to facilitate learning. The time, cost and effort required to create robust VPs on an individual school basis are significant; sharing of VPs by medical schools allows for access to a broad range of VPs across a variety of disciplines with lower investment. When this digital content is shared with other schools and distributed widely, digital copyright issues come into play. Unless all intellectual property rights (IPRs) and plans of the authors regarding the VP are confirmed upfront, the ability of the school to share the VP may be inhibited. Schools should also identify under what licensing/sharing model they plan to distribute the VPs - how do you plan to share the VPs and what will allow users to do with the VPs in the context of IPRs? This article highlights the role of IPRs in VPs and discusses a case-study of a European Virtual Patient collaboration to demonstrate how IPRs were managed. PMID- 19811207 TI - Virtual patients in a virtual world: Training paramedic students for practice. AB - Collaborative learning through case-based or problem-based learning (PBL) scenarios is an excellent way for students to acquire knowledge and develop decision-making skills. However, the process is threatened by the movement towards more self-directed learning and the migration of students from campus based to workplace-based learning. Paper-based PBL cases can only proceed in a single direction which can prevent learners from exploring the impact of their decisions. The PREVIEW project, outlined in this article, trialled a replacement to traditional paper PBL with virtual patients (VPs) delivered through a virtual world platform. The idea was that an immersive 3D environment could provide (a) greater realism (b) active decision-making and (c) a suitable environment for collaboration amongst work-based learners meeting remotely. Five VP scenarios were designed for learners on a Paramedic Foundation Degree within the virtual world second life (SL). A player using the MedBiquitous VP international standard allowed cases to be played both within SL and on the web. Three testing days were run to evaluate the scenarios with paramedic students and tutors. Students unfamiliar with the SL environment worked through five PBL scenarios in small groups, shadowed by 'in-world' facilitators. Feedback indicated that the SL environment engages students effectively in learning, despite some technology barriers. Students believed SL could provide a more authentic learner environment than classroom-based PBL. PMID- 19811208 TI - Online virtual patients - A driver for change in medical and healthcare professional education in developing countries? AB - The development of online virtual patients has proved to be an effective vehicle for pedagogical and technological skills transfer and capacity building for medical and healthcare educators in Malawi. A project between the University of Edinburgh and the University of Malawi has delivered more than 20 collaboratively developed, virtual patients, contextualised for in-country medical and healthcare education and, more significantly, a cadre of healthcare professionals skilled in developing digital resources and integrating these into their emerging curricula. The process of engaging with new approaches to teaching and delivering personalised, context sensitive content via a game-informed, technology-supported process has contributed to the ability of healthcare educators in Malawi to drive pedagogical change, meet the substantial challenges of delivering new curricula, cope with increasing student numbers and promote teacher professional development. This initial phase of the project has laid the foundation for a broader second phase that focuses on promoting curriculum change, developing educational infrastructure and in-country capacity to create, and integrate digital resources into education and training across multi-professional groups and across educational levels. PMID- 19811209 TI - Practica continua: Connecting and combining simulation modalities for integrated teaching, learning and assessment. AB - Simulation modalities are generally used independently of one another, largely due to physical and operational limitations to integration. Recent developments are enabling simulators and simulation environments to progress beyond single intervention models towards integrated continua of simulation. Moving to greater integration can improve contextualisation, better management of the transition from individual simulation to clinical practice, and provide wider opportunities to synthesise skills and approaches to practice. Simulation integration may involve experiential, modelling, technical, narrative, and evaluation dimensions; it includes both direct actions and activities, and technical and systems designs. The work in developing these integration continua is ongoing and takes many forms in many places. The framework of 'practica continua' proposed in this article links theoretical approaches and practical examples of integrated uses of simulation in education. PMID- 19811210 TI - Cross-cultural use and development of virtual patients. AB - Three major issues drive the cross-cultural use of virtual patients (VPs): an increased mobility of healthcare professionals, students and patients; limited resources for developing VPs; and emerging standards for the exchange of VPs across institutions. Many students are trained in countries other than where they were born. In addition, healthcare professionals often move between countries and are today meeting more and more patients from cultures different from their own. VPs can be used both for learning a new "medical" language as well as for illustrating different perspectives on illness in the new culture. Therefore, it may be important to develop cases reflecting patients from a wide variety of regions and cultures to prepare these professionals to understand both the background of these patients as well as the different medical conditions they may present. However, the benefits of using VPs may be limited at many universities by insufficient resources to develop all the VPs needed for their curricula. The option to acquire VPs from other universities may therefore be appealing, but as these may only be available in English, it is important to consider whether VPs reflecting the local illness panoramas and medical procedures are needed. PMID- 19811211 TI - The use of virtual patients to assess the clinical skills and reasoning of medical students: initial insights on student acceptance. AB - BACKGROUND: Web-based clinical cases ("virtual patients", VPs) provide the potential for valid, cost-effective teaching and assessment of clinical skills, especially clinical reasoning skills, of medical students. However, medical students must embrace this teaching and assessment modality for it to be adopted widely. METHOD: We examined student acceptance of a web-based VP system, Web-SP, developed for teaching and assessment purposes, in a group of 15 second-year and 12 fourth-year medical students. RESULTS: Student acceptance of this web-based method was high, with greater acceptance in pre-clinical (second-year) compared with clinical (fourth-year) medical students. Students rated VPs as realistic and appropriately challenging; they particularly liked the ability of VPs to show physical abnormalities (such as abnormal heart and lung sounds, skin lesions, and neurological findings), a feature that is absent in standardized patients. CONCLUSIONS: These results document high acceptance of web-based instruction and assessment by medical students. VPs of the complexity used in this study appear to be particularly well suited for learning and assessment purposes in early medical students who have not yet had significant clinical contact. PMID- 19811212 TI - Towards a typology of virtual patients. AB - INTRODUCTION: Although on-screen "virtual patients (VPs)" have been around for decades it is only now that they are entering the mainstream, and as such they are new to most of the medical education community. There is significant variety in the form, function, and efficacy of different VPs and there is, therefore, a growing need to clarify and distinguish between them. This article seeks to clarify VP concepts and approaches using a typology of VP designs. METHODS: The authors developed a VP design typology based on the literature, a review of existing VP systems, and their personal experience with VPs. This draft framework was refined using a Delphi study involving experts in the field, and was then validated by applying it in the description of different VP designs. RESULTS: Nineteen factors were synthesized around four categories: general (title, description, language, identifier, provenance, and typical study time); educational (educational level, educational modes, coverage, and objectives); instructional design (path type, user modality, media use, narrative use, interactivity use, and feedback use); technical (originating system, format, integration, and dependence). CONCLUSION: This empirically derived VP design typology provides a common reference point for all those wishing to report on or study VPs. PMID- 19811213 TI - Creation of virtual patients from CT images of cadavers to enhance integration of clinical and basic science student learning in anatomy. AB - OBJECTIVE: The goal of this study was to determine whether computerized tomographic (CT) images of cadavers could be used in addition to images from patients to develop virtual patients (VPs) to enhance integrated learning of basic and clinical science. METHODS: We imaged 13 cadavers on a Siemens CT system. The DICOM images from the CT were noted to be of high quality by a radiologist who systematically identified all abnormal and pathological findings. The pathological findings from the CT images and the cause of death were used to develop plausible clinical cases and study questions. Each case was designed to highlight and explain the abnormal anatomic findings encountered during the cadaveric dissection. A 3D reconstruction was produced using OsiriX and then formatted into a QuickTime movie which was then stored on the Tufts University Sciences Knowledgebase (TUSK) as a VP. RESULTS AND CONCLUSIONS: We conclude that CT scanning of cadavers produces high-quality images that can be used to develop VPs. Although the use of the VPs was optional and fewer than half of the students had an imaged cadaver for dissection, 59 of the 172 (34%) students accessed and reviewed the cases and images positively and were very encouraging for us to continue. PMID- 19811214 TI - The replacement of 'paper' cases by interactive online virtual patients in problem-based learning. AB - St George's University of London (SGUL) has a Problem-Based Learning (PBL) curriculum for its undergraduate medicine course, using traditional paper-based patient cases. To counter the limitation that paper cases are linear and do not allow students to explore the consequences of decisions, interactive online virtual patients (VPs) were developed which allowed students to consider options as the cases unfold, and allow students to explore the consequences of their actions. A PBL module was converted to VPs, and delivered to 72 students in 10 tutorial groups, with 5 groups each week receiving VPs with options and consequences, and 5 groups receiving online VPs but without options. A comprehensive evaluation was carried out, using questionnaires, and interviews.Both tutors and students believed that the ability to explore options and consequences created a more engaging experience and encouraged students to explore their learning. They regretted the loss of paper and neither group could see any value in putting cases online without the options. SGUL is now adapting its transitional year between the early campus years and the clinical attachment years. This will include the integration of all technology-based resources with face-to-face learning and create a more adaptive, personalised, competency-based style of learning. PMID- 19811215 TI - Improving assessment with virtual patients. AB - Assessments should accurately predict future performance in a wide variety of settings yet be feasible to conduct. In medical education a robust and comprehensive system of assessment is essential to protect the public from inadequate professionals. The parameters for devising such an assessment are well defined, and good practice for writing examinations well-established. However even excellent written assessments are limited in their predictive validity, and limited in sampling, face and construct validity. The increasing availability and power of computing has led to growing interest in computer simulations for use in examinations, creating assessment virtual patients (AVPs). They can potentially test knowledge and data interpretation, incorporate images, sound or video and test decision making. Such AVPs could represent the most comprehensive, integrated assessment possible that is both objective and feasible. This article focuses on AVP design, distinguishing between linear and branched models, choice and consequence driven designs. It reviews the use of AVPs in the context of assessment theory. It presents different AVP designs discussing their benefits and problems. AVPs can become valuable components in high stakes medical exams, particularly in later years of courses. However this requires application of established assessment principles to AVP design. PMID- 19811216 TI - Training staff to create simple interactive virtual patients: the impact on a medical and healthcare institution. AB - BACKGROUND: Virtual patients (VPs) are excellent teaching tools for developing clinical decision-making skills and improving clinical competency, but are believed to be very expensive and time consuming to make. AIM: The aim of this study was to establish whether it was possible to design a workshop for VP creation, which would enable teaching staff to create interactive, immersive VPs quickly, and with limited technical support. METHODS: The Centre for Medical and Healthcare Education at St George's University of London's (SGUL) medical school developed an ergonomic and generic 'model' for VP creation, simple enough for clinicians and educators to use, yet flexible enough to simulate real decisions through non-linear pathways. One-day workshops were set up to support the development of VPs by medical and healthcare educators. RESULTS: VP creation workshops have been successfully trialled, attracting a large number of clinicians and educators from a range of medicine and healthcare courses. Feedback from participants was very positive. Educators, organised into small groups, were unable to complete VPs within the workshop, but many groups completed a VP after the workshop. Interest was highest in mental health. DISCUSSION: The workshops catalysed a change in the awareness of the value of VPs, with staff directly integrating VPs into the curriculum. PMID- 19811217 TI - Educational institutions' Information Technology policies have the potential to impede e-learning research. PMID- 19811219 TI - Assessing symptoms of hypogonadism by self-administered questionnaire: qualitative findings in patients and controls. AB - BACKGROUND: Current screening instruments for hypogonadism lack adequate specificity and diagnostic accuracy. A new self-administered questionnaire of hypogonadism symptoms is being developed to address this need. The process for questionnaire development and results from the first (qualitative) phase are presented. METHODS: Qualitative interviews were conducted based on a new conceptual model of hypogonadism and according to standards for questionnaire development. An item pool was generated from focus groups and in-depth interviews with two groups of hypogonadal patients, treated (N = 26) and untreated (N = 26), and age-equivalent controls (N = 28). Standardized scoring of the qualitative interviews was used to confirm conceptual domains in the model and to generate questionnaire items for further validation. RESULTS: Key domains identified in both patients and controls included: (a) physical function; (b) bodily signs and symptoms; (c) sexual function and libido; (d) sleep function; (e) mood and affective function; (f) memory and cognitive function. The final domain is distress or bother associated with hypogonadism symptoms. This domain was only relevant to the patient groups. CONCLUSIONS: The first stage in the design of a new hypogonadism screener has been completed. Seven domains were identified and draft items were developed in each domain according to current standards of patient-reported outcomes. PMID- 19811220 TI - Microparticles in deep venous thrombosis, antiphospholipid syndrome and Factor V Leiden. AB - Microparticles (MPs) are blebs released from cellular surfaces during activation/apoptosis. They are procoagulant, pro-inflammatory and could contribute to pathogenesis of deep venous thrombosis (DVT). This study compared the number, cellular origin and procoagulant activity of MPs on DVT patients in different clinical situations: at diagnosis (n = 9, 5F/4M; mean age = 41.11), 1-3 years after warfarin withdrawal (n = 10, 7F/3M; mean age = 32.90), associated to antiphospholipid syndrome (APS; n = 11, 9F/2M; mean age = 33.82), or asymptomatic carriers of Factor V Leiden (FVL; n = 7, 7F/0M; mean age = 34.00) vs healthy controls (CTR). The quantification and characterization were performed by flow cytometry using CD235, CD61, CD45, CD31, CD14, CD45, anti-TF and Annexin V. The plasmatic procoagulant activity was investigated by prothrombin fragment 1 + 2 (F1 + 2) determination. The MPs procoagulant activity was analyzed by D-dimer (DD2) and Thrombin Generation Test (TGT) on a healthy pool of plasmas adjusted or not by their number (10,000 MPs). The MPs percentages were not different between the groups, but absolute number was increased in patients 1-3 years after warfarin withdrawal vs CTR (P = 0.02). There was no difference of the MPs cellular origin comparing patients to controls. TGT using 10,000 MPs was lower on these patients (P = 0.01). APS patients showed a reduction of plasmatic procoagulant activity (P = 0.004), but they were under warfarin therapy. DD2 in the presence of MPs, independently of its number, was higher in patients with DVT at diagnosis (P < 0.0001). MPs of patients with spontaneous DVT at diagnosis can promote coagulation activation demonstrated by increased DD2. Even the increased MPs from patients 1-3 years after thrombotic episode generated lower amount of thrombin, they can have a protective effect by activation of Protein C anticoagulant pathway. PMID- 19811221 TI - Metabolism of 2-acylglycerol in rabbit and human platelets. Involvement of monoacylglycerol lipase and fatty acid amide hydrolase. AB - The endocannabinoids 2-arachidonoylglycerol (2-AG) and anandamide (N arachidonoylethanolamine, AEA) are produced by neurons and other cells, including platelets, in a stimulus-dependent manner and act as signaling molecules; they are then inactivated through transport into cells followed by enzymatic degradation. A number of studies showed that monoacylglycerol lipase (MAGL) plays an important role in the degradation of 2-AG. In this study we investigated the enzymatic degradation of 2-acylglycerols in rabbit platelets and we characterized the responsible enzyme(s). [(3)H]2-AG and [(3)H]2-oleoylglycerol (2-OG) were both metabolized to [(3)H]glycerol and the respective fatty acid in a time and protein concentration-dependent manner, apparently by the action of MAGL activity. In the presence of the specific fatty acid amide hydrolase (FAAH) inhibitors URB597 and AM374, though, 2-OG hydrolysis was inhibited up to 55% in a concentration dependent manner (IC(50) = 129.8 nM and 20.9 nM respectively). These results indicate the involvement of both MAGL and FAAH on 2-acylglycerol hydrolysis. MAGL was further characterized in the presence of URB597 and it was found that 2 monoacylglycerols were hydrolyzed in a time, pH and protein concentration dependent manner and hydrolysis followed Michaelis-Menten kinetics, with an apparent K(M) of 0.11 microM and V(max) of 1.32 nmol/min*mg protein. Subcellular fractionation of platelet homogenate showed that MAGL activity was present in both the cytosolic and membrane fractions. In conclusion, the endocannabinoid 2 AG, as well as other 2-acylglycerols, are substrates of both FAAH and MAGL; the latter was characterized for the first time in platelets. In human platelets, under the same experimental conditions, the hydrolysis of 2-acylglycerols was higher and MAGL activity showed a different sensitivity against the inhibitors mentioned above. Finally, immunoblot analysis revealed the presence of MAGL, both in rabbit and human platelets, with a molecular mass of approximately 33 kDa. PMID- 19811222 TI - Effects of clopidogrel on "aspirin specific" pathways of platelet inhibition. AB - The most widely accepted methods of assessing response to clopidogrel involve isolated ADP-induced platelet aggregation. Whilst poor response determined by these assays correlates with adverse clinical events, the number of "poor responders" is far higher than the number of events attributed to treatment failure. Clopidogrel may have effects that cannot be assessed using isolated ADP induced aggregation. We have investigated the effect of clopidogrel on Arachidonic Acid (AA) induced platelet activation-an "aspirin specific" pathway using a novel near patient assay. Thirty four volunteers on no medication and 36 patients, on maintenance therapy with aspirin 75 mg daily, were recruited. Blood tests for Thrombelastogram PlateletMapping were taken immediately prior to and 6 hours after administration of a 600 mg clopidogrel loading dose. Changes in the area under the response curve at 15 minutes (AUC15) with both ADP- and AA stimulation were calculated as were the corresponding percentage platelet and percentage clotting inhibition (%PIn and %CIn). There were predictable and significant changes in the AUC15 of the ADP channel in response to clopidogrel and the corresponding %PIn and %CIn in both volunteers and patients. There were also significant reductions in the AUC15 of the AA channel (presented as Mean +/- 95%CI), by 27.2 +/- 11.8%, p = 0.005 in volunteers and 35.0 +/- 8.2%, p < 0.001 in patients) and increases in the %PIn and %CIn calculated using the AA channel in volunteers (by 20.0 +/- 11.4%, p + 0.02 and 32.3 +/- 12.8%, p < 0.001 respectively) and patients (by 24.2 +/- 8.6%, p < 0.001 and by 18.0 +/- 8.6, p < 0.001 respectively). Clopidogrel has both independent and aspirin-synergistic effects on AA-induced platelet activation suggesting potentiation of the antiplatelet activity of aspirin. This effect may be clinically important and is not detected by current "gold standard" methods of assessing response to clopidogrel. PMID- 19811223 TI - MicroRNA expression profiling of megakaryocytes in primary myelofibrosis and essential thrombocythemia. AB - In primary myelofibrosis (PMF) and essential thrombocythemia (ET) the megakaryocytic lineage characteristically shows aberrant proliferation and maturation in which the regulatory microRNA (miR) system might be involved. Laser microdissected PMF and ET megakaryocytes were analysed with real-time polymerase chain reaction (PCR) low density arrays comprising 365 microRNAs. The highest megakaryocytic expression levels were observed for miR-223, which is known to be expressed also in granulopoiesis. Cluster analysis revealed a tendency of disease specific megakaryocytic microRNA expression profiles indicating that a complex shift of microRNA expression appears to be the underlying defect. Increased accumulation of miR-146b was observed in cellular stage PMF (p = 0.0125) but not ET megakaryopoiesis. In conclusion, this is the first microRNA profiling of in situ-derived PMF, ET and normal megakaryocytes. PMID- 19811224 TI - Platelet-rich plasma in combination with bovine derived xenograft in the treatment of deep intrabony periodontal defects: a report of 20 consecutively treated patients. AB - There is currently great interest concerning the use of platelet-rich plasma (PRP) in combination with bone grafts for predictably obtaining periodontal regeneration. The aim of the present study was to investigate the effectiveness of PRP and bovine derived xenograft (BDX) combination in the treatment of deep intrabony defects with an emphasis on the evaluation of early wound healing. A total of 85 intrabony defects with an intrabony component of > or =3 mm were selected in 20 advanced chronic periodontitis patients. Defects were surgically treated with PRP/BDX. At baseline and 12 months after surgery, the following parameters were recorded: plaque and sulcus bleeding indices, probing depth (PD), relative attachment level, marginal recession (REC), probing bone and radiographic bone levels. Postoperative healing was evaluated by an early healing index at 1 and 2 weeks after surgery. At 12 months, all clinical and radiographic parameters were improved (p < 0.0001). The mean changes at 12 months were: PD reduction of 4.78 +/- 1.20 mm, attachment gain of 4.24 +/- 1.03 mm, REC of 0.54 +/- 0.34 mm, clinical bone gain of 3.75 +/- 0.97 mm, and radiographic bone gain of 3.79 +/- 1.02 mm, respectively. Two weeks after surgery, primary closure was maintained in 95% of the defect sites. Treatment with a combination of PRP and BDX leads to a significantly favorable clinical and radiographic improvement in deep intrabony periodontal defects. PMID- 19811225 TI - Preliminary evidence for a matrix metalloproteinase-2 (MMP-2)-dependent shedding of soluble CD40 ligand (sCD40L) from activated platelets. AB - Platelets are the major source of soluble CD40 ligand (sCD40L) in the blood. It has been demonstrated that CD40L is cleaved from the surface of activated platelets to release sCD40L. However, the enzyme involved in sCD40L shedding has not been identified yet. Using a panel of pharmacological inhibitors of serine, cysteine, aspartate, or metalloproteinases, preliminary evidence is presented for the hypothesis that matrix metalloproteinase-2 (MMP-2) might be the protease, primarily responsible for CD40L cleavage from platelet surface. PMID- 19811226 TI - New trends in hormone therapy. PMID- 19811228 TI - IMS consensus statements - 'what is their role?'. PMID- 19811230 TI - Acupuncture for hot flushes. PMID- 19811229 TI - Aging, menopause, cardiovascular disease and HRT. International Menopause Society Consensus Statement. PMID- 19811231 TI - Menopause and hormone replacement. Introduction. PMID- 19811232 TI - Incidence and mortality of cardiovascular disease in postmenopausal women world wide and relevance for preventive strategies. AB - World-wide, cardiovascular disease is mainly seen as a male problem, whereas breast cancer is considered to be the most important chronic disease in women. However, cardiovascular disease is the largest single cause of death also in women, accounting for approximately one-third of the mortality, while breast cancer is responsible for less than 5% of the mortality. Physicians and women are equally unaware of these numbers. From a public health viewpoint, morbidity might be even more important, given the fact that therapeutic strategies have improved in the last decades, and disease fatality has been decreasing. In this contribution, the mortality and disability-adjusted life-years of cardiovascular disease world-wide will be summarized, and the relevance for prevention and the specific issues relevant to postmenopausal women will be discussed. PMID- 19811233 TI - Mechanisms of action of estrogen receptors in vascular cells: relevance for menopause and aging. AB - Estrogen exerts pleiotropic functions on the cardiovascular system through binding to estrogen receptors (ERs). Traditionally, ERs have been recognized as transcription factors regulating the expression of target genes. In the past decades, however, numerous studies have revealed rapid actions of estrogen in different systems, especially in non-reproductive tissues such as the cardiovascular system. At this level, estrogen triggers rapid vasodilatation, exerts anti-inflammatory effects, regulates vascular cell growth and migration, and confers protection to cardiomyocytes. These so-called 'extranuclear actions' do not require gene expression or protein synthesis and are independent of the nuclear localization of ERs. Indeed, some of these actions are elicited by ERs residing at or near the plasma membrane. Through complex interactions with membrane-associated signaling molecules such as ion channels, G proteins and the tyrosine kinase c-Src, liganded extranuclear ERs lead to the activation of downstream cascades such as mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-OH kinase (PI3K). These cascades are responsible for important cardiovascular actions of estrogen, for instance, the activation of nitric oxide synthesis or the remodeling of the endothelial actin cytoskeleton. Moreover, these cascades play crucial roles in regulating the expression of target proteins implicated in cell proliferation, apoptosis, differentiation, movement and homeostasis. Recent advancements in the characterization of the molecular basis of the extranuclear signaling of estrogen help us to understand the biological functions of estrogen and would be beneficial in elucidating current controversies on estrogen's clinical efficacy in the cardiovascular system. PMID- 19811234 TI - Estrogen receptor actions on vascular biology and inflammation: implications in vascular pathophysiology. AB - Whereas hormonal therapy (HT) may increase the risk of coronary heart disease (CHD) and stroke in menopausal women, epidemiological studies (protection in premenopausal women) suggest and experimental studies (prevention of fatty streak development in animals) demonstrate a major atheroprotective action of estradiol (E2). The understanding of the deleterious and beneficial effects of estrogens is thus required at both a cellular and molecular level. Both the endothelium and the immuno-inflammatory system play a key role in the development of fatty streak deposit as well as in the rupture of the atherosclerotic plaque. Whereas E2 favors an anti-inflammatory effect in vitro (cultured cells), it rather elicits a pro-inflammatory response in vivo at the level of several subpopulations of the immuno-inflammatory system, which could contribute to plaque destabilization. E2 promotes beneficial actions on the endothelium such as nitric oxide and prostacyclin production. E2 actions are essentially mediated by two molecular targets: estrogen receptor alpha (ER-alpha) and beta (ER-beta), but the former appears to mediate most of the actions of E2 on the endothelium and on the immune system. ER-alpha modulates target gene transcription through two activation functions (AF), AF-1 and AF-2, even though signalling via ER-alpha located at the plasma membrane (responsible for membrane-initiated steroid signalling (MISS)/(extra-genomic)) can also lead to an indirect effect on gene transcription. Recently, we demonstrated that ER-alpha AF-1 is not required for the vasculoprotective actions of E2, whereas it is necessary for the effects of E2 on its reproductive targets. These results suggest that selective estrogen receptor modulators stimulating ER-alpha with minimal activation of ER-alpha AF-1 could retain beneficial vascular actions, while minimizing the sexual effects. PMID- 19811235 TI - Estrogen actions in the cardiovascular system. AB - This brief review summarizes the current state of the field for estrogen receptor actions in the cardiovascular system and the cardiovascular effects of hormone replacement therapy (HRT). It is organized into three parts: a short Introduction and overview of the current view of how estrogen works on blood vessels; a summary of the current status of clinical information regarding HRT and cardiovascular effects; and an update on state-of-the-art mouse models of estrogen action using estrogen receptor knockout mice. PMID- 19811236 TI - Cardiovascular risk and events in polycystic ovary syndrome. AB - Young women with polycystic ovary syndrome (PCOS) present a high risk for cardiovascular disease because of the presence of abdominal obesity, insulin resistance and androgen excess. In addition, they present with endothelial dysfunction and early signs of atherosclerosis (increased carotid intima-media thickness and increased coronary calcium). However, the evidence of increased cardiovascular events during the postmenopausal age is relatively small, although some recent studies have indicated a slight increase in the severity of cardiovascular disease in women who had PCOS during their fertile age. The discrepancy between cardiovascular risk in young age and postmenopausal cardiovascular events may depend on changes in the risk that happen during the late fertile age or on overestimation of early atherosclerotic signs. PMID- 19811237 TI - Premature menopause increases cardiovascular risk. AB - Premature menopause and bilateral oophorectomy in young women are associated with an increased incidence of cardiovascular disease, myocardial infarction and overall mortality. Observational studies suggest an interval of 5-10 years between loss of ovarian function and the increased risk of cardiovascular disease. This finding is consonant with a published autopsy study of women who had undergone bilateral oophorectomy. The progression of atherosclerosis is retarded with the use of estrogen replacement therapy in non-human primates and women. Hormone therapy reduced the incidence of cardiovascular disease in women following bilateral oophorectomy. These findings support the use of hormone therapy in young women who have lost ovarian function. PMID- 19811238 TI - Vasomotor symptoms and cardiovascular risk. AB - Climacteric complaints are the main indication for hormone replacement therapy (HRT) in the clinical practice. Observational studies demonstrating a protective effect of HRT on cardiovascular disease (CVD) were conducted in early menopausal, young, symptomatic women. Vasomotor symptoms correlate with lower level of plasma antioxidant activity, an increased cardiovascular reactivity to stressful situations, elevated cholesterol, higher sympathetic nerve activity, impaired flow-mediated dilation, hypertension and a higher risk of aortic calcification. All the available findings indicate that hot flushes can be seen as a marker for underlying vascular changes among mid-life, otherwise healthy, climacteric women. Thus, young, healthy symptomatic postmenopausal women differ from those without vasomotor symptoms with regard to cardiovascular risk factors. Therefore, responses to HRT can change in terms of cardiovascular outcomes according to the baseline vasomotor complaints. This point may explain, at least in part, the negative/null effects of HRT on cardiovascular disease observed in the trials where HRT was given to largely asymptomatic, elderly women. PMID- 19811239 TI - Blood pressure through aging and menopause. AB - Together with the aging process, hypertension is the main risk factor contributing to the increase in cardiovascular morbidity and mortality in postmenopausal women, with a prevalence of around 60% in women older than 65 years. Considering that hypertension is a modifiable risk factor, the understanding of its epidemiology and pathophysiology and the development of appropriate therapeutic strategies are conceivably crucial in reducing cardiovascular risk. The high prevalence of hypertension in older women is largely due to the progressive stiffening of the arterial structure which accompanies the aging process in both sexes. However, the abrupt fall in circulating estrogen levels might independently contribute to the rise in blood pressure, through partly unknown mechanisms, such as a direct effect on the arterial wall, the activation of the renin-angiotensin system and of the sympathetic nervous system. Postmenopausal hypertension fosters the development of left ventricular hypertrophy and is the main factor contributing to coronary artery disease, chronic heart failure and stroke in older women. Recent analysis demonstrates that men and women receive a similar benefit from antihypertensive therapy in terms of reduction of cardiovascular morbidity and mortality and, considering that generally the response to different drugs is not different between the sexes, currently there is no need to use specific antihypertensive drug classes after menopause. Finally, although observational studies have shown that hormone replacement therapy is associated with lower cardiovascular risk and lower blood pressure values, randomized clinical trials have conversely denied this benefit and demonstrated rather that this therapeutical approach increases the risk of cardiovascular events. PMID- 19811240 TI - Cardiovascular aspects of menopausal hormone replacement therapy. AB - Cardiovascular disease is the leading cause of death in postmenopausal women in Western countries. Despite preventive strategies, in the past decades, the incidence of cardiovascular events has shown a decline in men but a rise in women, matching the growth of the population of postmenopausal women. Several epidemiological findings suggest the causative role of ovarian hormone deficiency in the development of cardiovascular disease in women. Observational and randomized studies have suggested that initiation of hormone replacement therapy in early postmenopause could be beneficial from a cardiovascular point of view. Conversely, aging, time since menopause and presence of cardiovascular risk factors or cardiovascular disease may decrease its efficacy and increase the risk of cardiovascular events. It is plausible that the unfavorable effects of the estrogen/progestin combination used in the randomized studies are not related to the hormone preparation per se but rather to the use of hormones in the less receptive group of women, older and with cardiovascular risk factors. Clinical judgement, choice of the right dose and estrogen/progestin combination are of pivotal importance to maximize the beneficial effect of estrogen or estrogen progestin replacement therapy, especially if given within a reasonably short time after the menopause to those women that need this therapy for the relief of menopausal symptoms. In these women, continuation of estrogen or estrogen progestin replacement therapy may be beneficial. PMID- 19811241 TI - The hemostatic system through aging and menopause. AB - The process of aging is accompanied by several modifications in the hemostatic system at different levels (blood coagulation, fibrinolysis, platelet activity, vascular endothelium). These changes may explain the higher incidence of arterial and venous thrombosis in the elderly compared to young people. Genetic and environmental factors modulate in different combinations the expression of proteins involved in the hemostatic process. Among the latter, diet and smoking habits play an important role, as well as physical exercise and, for women, hormonal status. A gradual and progressive development of a low-grade inflammatory state (clearly demonstrated in the elderly) is also an important factor that influences hemostasis during aging. In spite of the fact that the increased hypercoagulable state observed with aging may account for the higher incidence of thrombosis in the elderly, the finding of a similar pattern of coagulation activation in healthy centenarians suggests that a hypercoagulable state is compatible with health and longevity. Taking also into consideration that no laboratory parameters of hemostasis are predictive of thrombosis on an individual basis, a physician's behavior towards aging patients (e.g. prescription of hormonal replacement therapy to a woman during menopause) should not be affected by laboratory tests, but mainly by a patient's clinical history and the presence of strong risk factors for thrombosis other than age (e.g. diabetes mellitus, arterial hypertension, dyslipidemia, obesity, smoking). PMID- 19811242 TI - Quality of life and sexuality issues in aging women. AB - Quality of life may decrease after menopause. Hormone replacement therapy remains the first-line and most effective treatment for menopausal symptoms and improvement of low quality of life due to estrogen deficiency. The decrease of health-related quality of life in women suffering from cardiovascular disease may be superimposed on the decrease of quality of life induced by menopause itself. Postmenopausal women with acute cardiovascular disease have a significantly higher probability of death than men of the same age. Quality of life predicts long-term mortality. A myocardial infarction does not automatically interdict sexual activity. The Princeton guidelines classify patients suffering from cardiovascular diseases in three categories. Most patients belong to the low-risk category. In general, these patients can be safely encouraged to initiate or resume sexual activity or to receive treatment for sexual dysfunction. Patients at intermediate (or indeterminate) levels of risk should further receive cardiologic evaluation to be classified into either the low- or high-risk group. Patients in the high-risk category have to be stabilized by specific treatment for their cardiac condition before resumption of sexual activity, or initiation of treatment for sexual dysfunction. PMID- 19811243 TI - Cardiovascular risk assessment in postmenopausal women: the role of the gynecologist. AB - The gynecologist is often the only physician a woman consults on a regular basis and therefore gynecologists play a crucial role in the primary prevention of cardiovascular disease. Cardiovascular risk factors in women entering the menopausal transition are poorly managed, partly due to the fact that primary care physicians, gynecologists and cardiovascular physicians often fail to identify cardiovascular risk factors and also undertreat women at increased cardiovascular risk. Furthermore, most women are not well informed about their cardiovascular risk profile. Gynecologists and cardiologists should work together as a team in identifying and managing cardiovascular risk factors. European cardiologists and gynecologists have written a Consensus statement and a short guide to help menopause physicians to assess and manage the cardiovascular risk in women. PMID- 19811244 TI - Lifestyle and diet in postmenopausal women. AB - Diet and nutritional habits have a pivotal role in maintaining human health. Unhealthy eating, obesity and nutritional deficiencies may lead to various diseases. However, a most important component of lifestyle relates to physical activity. Sedentary people fare less well than those who exercise regularly. The benefits of exercise can be demonstrated in many bodily organs. The most frequently studied effect of exercise is reduction in cardiovascular morbidity and mortality, but positive effects on the musculoskeletal system, breast cancer risk, mood and cognition, and quality of life have been recorded as well. In many cases, a dose-response was evident, and even a mild to moderate degree of activity, performed only a few times weekly, may carry significant merits. The following article reviews this topic and brings updated information on dietary advice and the benefits of exercise in postmenopausal women. PMID- 19811245 TI - Comparative effects of conventional vs. novel hormone replacement therapy on blood pressure in postmenopausal women. AB - Menopause is commonly characterized by an increase in blood pressure. Higher blood pressure may partially explain the elevated risk for cardiovascular events observed in postmenopausal women. There is a graded relationship between blood pressure and cardiovascular risk which extends to levels of blood pressure well below 140/90 mmHg, the classical established blood pressure limit for the diagnosis of hypertension. Despite this knowledge and the wide availability of consensus treatment guidelines for hypertension, high blood pressure remains untreated or poorly treated in many postmenopausal patients. It is clear that novel and innovative population strategies for lowering blood pressure in postmenopausal women are warranted. Clinical trials suggest that oral estrogen administration may produce either a neutral effect or a small increase in blood pressure in postmenopausal women. Transdermal estrogen administration has not been studied as extensively but may produce a decrease in blood pressure. The mechanisms for the differences observed between oral and transdermal estrogen have not been completely elucidated. Drospirenone (DRSP) is a novel progestin with aldosterone receptor antagonist activity that has been developed for hormone therapy as DRSP/17beta-estradiol (DRSP/E2). In several clinical trials, DRSP/E2 has demonstrated a significant antihypertensive effect as well an additive effect when combined with existing antihypertensive therapy. Despite the wide availability of treatment guidelines, a wide array of antihypertensive agents, and the introduction of hormone replacement therapy that can lower blood pressure, optimizing blood pressure control remains a serious issue confronting the physician who cares for the postmenopausal woman. PMID- 19811246 TI - Coronary heart disease and hormone replacement therapy after the menopause. AB - The discordance in coronary heart disease (CHD) outcome between randomized, controlled trials and observational studies of hormone replacement therapy (HRT) is likely the result of the dissimilar cohorts studied. This observation led to the formation of the timing hypothesis that benefits and risks of HRT depend upon age of HRT initiation and/or time of HRT initiation in relation to menopause. This hypothesis has been supported with data from large, randomized, controlled trials that have studied HRT and selective estrogen receptor modulators (SERMs) in the prevention of CHD. Initiation of HRT in women <60 years of age and/or within 10 years of menopause reduces both CHD and total mortality; SERMs have been shown to reduce CHD in women <60 years. What has become clear from the cumulated literature is that in young postmenopausal women who initiate HRT in close proximity to menopause, the adverse effects of HRT are rare and no greater than those of other commonly used pharmacological agents and that the primary prevention benefits for CHD are at least equivalent to those of other commonly used therapies. Additionally, the literature indicates that the duration of HRT confers greater CHD benefit especially in women <60 years who initiate therapy. The cumulated literature dispels misperceptions concerning HRT and CHD. PMID- 19811247 TI - Hormone therapy and risk of venous thromboembolism among postmenopausal women. AB - Venous thromboembolism, either deep vein thrombosis or pulmonary embolism, is a serious side-effect of postmenopausal hormone therapy. Current use of oral estrogens increases the risk of venous thromboembolism, especially during the first year of treatment, but past users of hormone therapy have a similar risk as never-users. Among women at high risk for venous thromboembolism (for example, thrombogenic mutations, obesity), oral estrogens use further enhances the thrombotic risk. Recent studies have shown that transdermal estrogens might be safe with respect to thrombotic risk. The difference in thrombotic risk between oral and transdermal estrogens may be partially explained by changes in hemostasis. Few data are currently available regarding the impact of progestogens on venous thromboembolism risk, but norpregnane derivatives might be thrombogenic. Individual assessment of the benefit-risk ratio is needed before initiating treatment and oral estrogens should be avoided among women at high risk for venous thromboembolism. PMID- 19811248 TI - The risk of stroke in postmenopausal women receiving hormonal therapy. AB - Stroke affects one in five women and is the third leading cause of death in the United States. The incidence increases with age, and the number of females experiencing a stroke is similar to the number of men, unlike in coronary disease, where the number of females suffering from the disease is lower before menopause and then slowly becomes equal to the number of men. There are multiple risk factors for stroke, but the most significant ones influencing the relationship between hormones and stroke are obesity, hypertension and smoking. There have been multiple studies looking at the relationship between hormones and stroke. Among observational studies, 21 have shown no effect, while six have shown a decreased risk and four studies, an increased risk. It is clear that the risk of stroke with hormones is only for ischemic and not hemorrhagic stroke. Among randomized trials, the Women's Health Initiative data, because of the study's size, have provided the greatest weight in meta-analytic reviews. Overall, an increase of 20-40% has been found, and, because of various confounding factors, it has been difficult to determine whether this is statistically significant, although several studies have confirmed this for all postmenopausal women treated. However, in younger women, aged 50-59 years, there is little evidence that the risk is significantly increased, and the overall background prevalence is low: 6-8/10 000 women/year. Recent studies have suggested the risk is not increased significantly with transdermal therapy and with lower doses, although use of progestogen does not appear to influence the risk assessment. There is lack of consistent data on whether early age of initiation of therapy affects the risk. Because this risk may pertain to younger postmenopausal women, and there is a very small risk of stroke reported for young women taking oral contraceptives, it is suggested that the small risk in younger women is through an inflammatory/thrombosis mechanism in susceptible women, rather than one of atherosclerosis, as with coronary disease. Nevertheless, in younger postmenopausal women, with a lower baseline prevalence rate, even if there is a 40% increased risk, the attributable risk would result in 1-2 additional cases/10 000 women/year, which would be considered a very rare event. PMID- 19811249 TI - Type and route of estrogen administration. AB - Hormone replacement therapy (HRT) can be administered orally and non-orally. Providing equivalent doses are given, all forms of HRT can equally relieve menopausal symptoms and prevent bone loss and osteoporosis. Different routes of administration will have differing metabolic effects, with oral HRT producing a hepatic first-pass effect not seen with non-oral HRT. The first-pass effect can produce benefits including larger reductions in low density lipoprotein cholesterol, lipoprotein(a) and insulin resistance, and larger increases in high density lipoprotein cholesterol. Unwanted effects are seen in increases in triglycerides and in coagulation activation. Cardiovascular effects of oral and transdermal HRT appear to be fairly similar, with improvements in vascular endothelial function, angiotensin-converting-enzyme activity, and in most markers of inflammation. There is a paucity of studies on the effects of transdermal HRT on cardiovascular outcomes, but the few data available suggest similar effects to oral HRT, and dose rather than route of administration is probably more important in this respect. Oral HRT may be preferred in women with evidence of insulin resistance, such as in metabolic syndrome or maturity-onset diabetes mellitus. Transdermal HRT may be preferred in women with coagulation disturbances. But, for the majority of women, personal preference should determine their choice of HRT route. PMID- 19811250 TI - Estrogen dose: the cardiovascular impact. AB - One of the main concerns from the large secondary and primary prevention cardiovascular trials such as the Heart and Estrogen/progestin Replacement Study and the Women's Health Initiative (WHI) study was that there appeared to be an increased risk of coronary events in the first years of usage. These findings were initially surprising bearing in mind that observational trials had previously shown benefit. Subsequent analyses have confirmed that the risks were primarily in the over-seventies age group. In the younger age groups (50-59 years), there was a trend towards reduction of risk and a significant reduction in all-cause mortality. Why did this occur? There are a number of possible reasons but, in the opinion of the author, the dose of estrogen was the overriding factor. Although 0.625 mg of conjugated equine estrogens is a standard dose for women in their early fifties, this was a relative overdose for women with an average age of initiation of 63 years (WHI). Inappropriately high doses of estrogen could have caused cardiovascular harm due to disturbances in thrombogenesis and vascular remodelling. Lower doses of hormone replacement therapy, or more accurately, more appropriately targeted doses for each age group, could avoid initial harm, thus allowing for long-term benefits to predominate. PMID- 19811251 TI - Different cardiovascular effects of progestins according to structure and activity. AB - Following the publication of the Women's Health Initiative study, the controversy was raised regarding the role of progestins in hormonal replacement therapy (HRT). Some of the most prescribed molecules, with partial androgenic or glucocorticoid activity, have been shown to oppose partially the beneficial effect of estrogens on surrogate markers of cardiovascular disease risk. Unfortunately, this concern has been directed towards progestins as a class effect, although striking differences exist among the types of molecules used. The synthetic progestins used in HRT have varying pharmacologic properties depending on the molecules from which they are derived, either testosterone or progesterone. Very small structural changes in these molecules may induce considerable difference in their effects on various targets and especially on the surrogate markers of cardiovascular disease risk, where some molecules may reverse the beneficial effects of estrogen. Natural progesterone and some of its derivatives such as the 19-norprogesterone molecules or the new molecules drospirenone, a potent antimineralocorticoid agent with a beneficial effect on blood pressure, and dienogest do not exert any androgenic effect and have no negative effect on the lipids or on the endothelial cells. Although it is likely that the new progestins may be neutral on the coronary disease risk, when administered to the younger postmenopausal woman, this has not as yet been documented by large, randomized, controlled trials. PMID- 19811252 TI - Androgen replacement therapy and cardiovascular function. AB - Androgens play a primary role in female physiopathology. The age-related decline in the production of ovarian and adrenal androgens may significantly affect women's health. Scanty studies have been designed to establish whether androgen deficiency might directly affect cardiovascular biology and whether it might be a risk factor for cardiovascular disease in women during reproductive aging. The present short review examines the effect of androgen on the cardiovascular system, with particular regard to dehydroepiandrosterone and testosterone replacement in postmenopausal women. PMID- 19811253 TI - Clinical issues regarding cardiovascular disease and selective estrogen receptor modulators in postmenopausal women. AB - Selective estrogen receptor modulators (SERMs) have offered the promise of reducing the burden of coronary artery disease (CAD) in postmenopausal women, based on the positive effects recorded on intermediate markers (blood lipids and markers of inflammation). The effects of raloxifene, bazedoxifene and lasofoxifene on cardiovascular endpoint markers are presented as reported in recent, randomized, controlled trials. Raloxifene failed to significantly lower the risk of CAD in postmenopausal osteoporotic women, without any effect on stroke or early harm, but doubling the risk of venous thromboembolism. The risk of CAD was lowered in a subgroup of patients at risk of CAD. In a large randomized, controlled trial with CAD as the primary endpoint in patients at risk of CAD, raloxifene failed to significantly reduce CAD, while significantly increasing the incidence of fatal stroke and venous thromboembolism. Bazedoxifene, in an osteoporosis trial, had similar effects on the cardiovascular system when compared to raloxifene. Lasofoxifene has only been studied in postmenopausal osteoporotic women in the PEARL trial. Lasofoxifene reduced the risk of coronary heart disease events as well as the risk of stroke, while the risk of deep vein thrombosis remained in line with other SERMs. These results will need to be confirmed in a study with primary cardiovascular endpoints. Until then, it is unlikely that SERMs will play a major role in strategies aimed at the prevention of coronary heart disease in postmenopausal women. PMID- 19811254 TI - Local estrogens for quality of life and sexuality in postmenopausal women with cardiovascular disease. AB - Urogenital aging and female sexual dysfunction (FSD) are significant problems following menopause. Estrogen decline is one of the key factors contributing to sexual functioning because of its crucial role for genital arousal (vasocongestion and lubrication) and other domains of the sexual response. Several common medical conditions, including cardiovascular disease (CVD), may interfere with women's sexual response across the aging process. FSD is one of the most common CVD-related quality-of-life complications with a major impact on patients' and their sexual partners' life. There is no evidence that FSD may represent an early indication of cardiovascular risk in postmenopausal women. In spite of the high prevalence, FSD remains largely under-recognized and sexual counseling is an important consideration for the proper management of postmenopausal women with CVD. Many local estrogen products are available (creams, tablets, suppositories, pessaries and rings) and are equally effective for treatment of vaginal atrophy. When a history of CVD is present, local estrogens may be safely used to treat urogenital atrophy with a significant improvement of sexual health and quality of life. PMID- 19811255 TI - Anti-nuscleosome antibody, a reliable indicator for lupus nephritis. AB - Systemic lupus erythematosus (SLE) is characterized by the presence of antinuclear antibodies (ANA). ANAs are aimed against native DNA, as well as histone proteins and small nuclear ribonucleoproteins, also could be targeted against the entire chromatin, consisted of nucleosomes. Herein, we assessed the frequency and the concentration of anti-nucleosome antibodies (aNS) in a group of 107 Hungarian patients with SLE. Moreover, correlations between these autoantibody concentrations and disease activity, as well as organ manifestations, mainly renal symptoms were evaluated. The occurrence of positive aNS, anti-dsDNA and anti-histone (aH) antibodies were 39.2, 28.0 and 47.6%, respectively. All the three autoantibodies were represented with significantly higher titers in patients with lupus nephritis. A positive correlation was found between the concentration of these autoantibodies and disease activity. The long disease duration (mean 8.5 years) and consequent low disease activity (mean disease activity index: 3.28) could be the reason for the relatively low occurrence of ANAs amongst patients. Besides anti-dsDNA, the assessment of aNS and aH antibodies could be useful in the diagnosis and follow-up of SLE. The authors discuss the possible role of aNS autoantibodies in the pathogenesis of lupus nephritis. PMID- 19811256 TI - Abnormal distribution of dendritic cells in (NZW x BXSB)F1 mice. AB - (NZW x BXSB)F1 mice (W/BF1 mice) have been reported to develop autoimmune diseases with aging. We have also reported that the number of dendritic cells (DCs) increases in the various organs, and that the B-cell response to LPS or interleukin-4 plus anti-mu increase with aging in W/BF1 mice. In the present experiment, we show that many DCs exist not only in the T-cell area but also in the B-cell area and the sinus in the spleen of aged W/BF1 mice, and that the coculturing of DCs from aged W/BF1 mice and B cells from disease-free young W/BF1 mice produces much more IgG and IgM than normal mice. These results suggest that an abnormal distribution of DCs and the interaction of DCs and B cells induce the hyperproduction of immunoglobulin in aged W/BF1 mice. PMID- 19811257 TI - Functional polymorphisms of DEFB1 gene in type 1 diabetes Brazilian children. AB - We analyzed three functional 5' un-translated region beta-defensin 1 (DEFB1) single nucleotide polymorphism (SNPs) in a group of 170 type 1 diabetes (T1D) patients. In order to evaluate the SNPs influence on the disease onset and the development of other autoimmune disorder, such as celiac disease (CD) and autoimmune thyroid disease (AITD), patients were stratified according to the presence of AITD, CD, and both AITD and CD. As control group, we studied 191 healthy children and adolescent not presenting a familiar historic of T1D, CD or AITD. DEFB1 SNPs were in Hardy-Weinberg equilibrium both in healthy controls and T1D patients, as well in the T1D patients stratified according to the presence of other autoimmune disorder(s). Allele, genotype, and haplotype frequencies of T1D patients globally considered were comparable to healthy controls ones. No evidence of any association of DEFB1 SNPs with the onset of AIDT, CD, and both AITD and CD on T1D patients was evidenced. Only a minor trend was found for an increased frequency of the - 20 G allele in T1D patients only presenting AITD vs. T1D patients not presenting AITD or CD, as well as an increase of those haplotypes comprising the - 20 G allele when compared with the GCA haplotype. We also evaluated the influence of functional DEFB1 SNPs on the age of T1D onset: no significant statistical conclusion was achieved. Further studies are envisaged, in order to elucidate the possible role of functional DEFB1 polymorphisms in the onset of TD1 and other autoimmune-related disorders. PMID- 19811258 TI - In vitro model of annexin A5 crystallization on natural phospholipid bilayers observed by atomic force microscopy. AB - Annexin A5 is a potent anticoagulant protein with a thrombomodulatory function. It is frequently mentioned with systemic inflammatory autoimmune disease, which share higher vulnerability to cardiovascular diseases. The protein has the ability to bind to membranes containing negatively charged phospholipids in a calcium-dependent manner. The potent anticoagulant properties of the protein are a consequence of this crystallization, which forms the lattice of annexin A5 over phospholipid surface, blocking its availability for coagulation reactions. Crystallization of annexin A5 has been proven on homogeneous synthetic phospholipids. However, the crystallization of annexin A5 on inhomogeneous, naturally derived phospholipid surfaces, in p3 and p6 crystal form, has now been reported for the first time. Atomic force microscopy was chosen for the observation of the crystallization of annexin A5 on different solid supported phospholipid bilayers. In this study model, the optimal results were obtained by using: 0.5 mg/ml lipid vesicles suspension (70% phosphatidylcholine, 30% phosphatidylserine) in HEPES buffer saline (HBS) with 2 mM CaCl(2), large unilamellar vesicles with sizes around 200 nm, 41 degrees C of phase transition temperature and 21 microg/ml of native annexin A5 in HBS with 2 or 20 mM CaCl(2). Results were evaluated by imaging and force measurements. Demonstration that native annexin A5 is able to spontaneously crystallize on naturally derived, inhomogeneous phospholipids is supporting the putative role of annexin A5 crystal structures as possible antithrombotic shield. This in vitro system is probably more appropriate for studying the pathogenetic role of antiphospholipid antibodies. PMID- 19811259 TI - Transplantation of NIT-1 cells with ectopic FADDdel-GFP expression for treatment of streptozotocin-induced diabetes. AB - Islet transplantation is considered a therapeutic option for type 1 diabetes (T1D). However, allorejection is one major barrier for the successful islet transplantation. In the present study, we have tested the feasibility of a deletion construct for Fas-associated death domain protein (FADD; without the death effecter domain) fused with green fluorescent protein (FADDdel-GFP) for blocking the Fas-FasL signaling pathway in prevention of transplanted beta cell destruction by allo-rejection in T1D. In vitro studies have shown that NIT-1 cells with ectopic FADDdel expression were resistant to cytokine-induced apoptosis and CTL-mediated lysis. Diabetic Balb/c mice reached normoglycemia promptly and gained weight after transplantation of NIT-1 cells with ectopic FADDdel-GFP expression. These recipients showed a significant longer survival time than that of recipients transplanted with NIT cells with ectopic GFP expression only. Our results together suggest that FADDdel could be a useful target for the improvement of islet transplantation for T1D. PMID- 19811260 TI - The infectious etiology of vasculitis. AB - Infectious agents have been implicated in the etiopathogenesis of various vasculitides via numerous and overlapping mechanisms including direct microbial invasion of endothelial cells, immune complex mediated vessel wall damage and stimulation of autoreactive B and/or T cells through molecular mimicry and superantigens. While the causative role of hepatitis B virus in polyarteritis nodosa and hepatitis C virus in mixed cryoglobulinemia is clearly established, evidence for the association of other infectious agents with vasculitis, including human immunodeficiency virus, parvovirus B19, cytomegalovirus, varicella zoster virus, Staphylococcus aureus, rickettsiaceae, Treponema pallidum and Borrelia burgdorferi, among numerous others, is accumulating. The spectrum of association of infectious agents; bacteria, viruses and parasites, with systemic vasculitides, will be reviewed herewith. PMID- 19811261 TI - Infection and thyroid autoimmunity: A seroepidemiologic study of TPOaAb. AB - Infectious agents have been implicated as triggers of autoimmunity. Prospective epidemiologic studies of infection with specific pathogens and the subsequent elevation of specific autoantibodies are difficult and costly to conduct. As a result, a solid body of evidence regarding this theoretically intriguing connection remains to be accrued. We studied term sera from 1807 pregnancies in 1591 women for whom IgG status for cytomegalovirus, Epstein-Barr virus, herpes simplex virus type 1, herpes simplex virus type 2, and/or Toxoplasma gondii was available from prior analyses. We tested the sera (masked regarding infectious status) for autoantibodies to thyroid peroxidase (TPOaAb) and then unmasked and linked them. Adjusted for other cofactors, prior infection with T. gondii was associated significantly with the elevation of TPOaAb, whereas seropositivity for other infections was not. Negative and positive findings for suspected triggers of autoimmunity should be reported to build the evidentiary basis needed to advance our understanding of the disease process. The positive association observed between prior infection with T. gondii and the elevation of TPOaAb is supported by an almost simultaneous study. These findings require further investigation. We believe that if T. gondii is in fact confirmed to trigger or enhance a TPOaAb response, the most likely mechanism involved is the bystander effect. PMID- 19811262 TI - The p85beta regulatory subunit of phosphoinositide 3-kinase has unique and redundant functions in B cells. AB - Phosphoinositide kinase (PI3K) is activated by various receptors on lymphocytes and regulates development, activation, and tolerance. Genetic ablation of PI3K function in T cells leads to the appearance of autoimmune disorders. In B cells, loss of the class IA regulatory subunit p85alpha causes a partial defect in B cell development and proliferation, whereas loss of p85beta alone causes no apparent changes in B cell function. Here we investigate further the consequences of p85beta deletion in B cells, in the presence or absence of p85alpha. We demonstrate that p85beta partially compensates for loss of p85alpha in B cell development and peripheral survival, with greater defects observed when both isoforms are absent. BCR-mediated AKT phosphorylation is partially reduced in p85alpha-deficient B cells and further diminished with concomitant loss of p85beta. Unexpectedly, loss of p85beta results in increased BCR-mediated proliferation and ERK phosphorylation. These results indicate that the p85beta regulatory isoform has partially overlapping functions with p85alpha in B cells as well as a unique role in opposing BCR responses. PMID- 19811263 TI - Selected angiogenic cytokines in systemic lupus erythematosus patients. AB - The objective of this study was to determine the serum concentration of angiogenic factors (vascular endothelial growth factor, VEGF; transforming growth factor beta, TGF-beta1; hepatic growth factor, HGF; basic fibroblast growth factor, bFGF; tumor necrosis factor alpha, TNF-alpha; soluble vascular endothelial growth factor receptor 1, sVEGF-R1; soluble vascular endothelial growth factor receptor 2, sVEGF-R2), the relationships among them and to assess the relation of their levels with the applied therapy in 48 females with systemic lupus erythematosus (SLE; 37 long-term treated +11 newly diagnosed). The control group consisted of 24 healthy women. A statistically significant increase of sVEGF-R2 and significant decrease of sVEGF-R1 were observed in the subgroup of newly diagnosed SLE patients as compared to the control subjects. No significant differences were found between serum angiogenic factors in the long-term treated subgroup and the control, the long-term treated subgroup and the newly diagnosed SLE patients after a 3-month treatment, and the subgroup of newly diagnosed SLE patients before therapy and after a 3-month treatment. The significant decrease in the serum of sVEGF-R2 was revealed in the subgroup treated for a long-time as compared to the subgroup of newly diagnosed untreated SLE patients. The analysis of relationships between serum concentration of sVEGF-R1 and other cytokines levels revealed positive correlation with concentration of VEGF and TNF-alpha in the total group of patients. In the newly diagnosed untreated subgroup, a strong positive correlation between concentration of sVEGF-R1 and bFGF was observed. Furthermore, a moderate positive correlation between concentration of sVEGF-R1 and the level of VEGF was revealed in the long-term treated patients. The association between sVEGF-R2 and HGF was also noted in this subgroup. The obtained data suggest the necessity of further investigations to determine the importance of angiogenic factors in pathogenesis and therapy of SLE. PMID- 19811264 TI - Retinoid X receptor beta polymorphisms do not explain functional differences in vitamins D and A response in Antineutrophil cytoplasmic antibody associated vasculitis patients. AB - It has been suggested that the retinoid X receptor beta (RXRB) gene is a risk factor for Wegener's granulomatosis. We addressed if there is a functional difference in the response to retinoic acid (RA) and vitamin D in Antineutrophil cytoplasmic antibody (ANCA) associated systemic vasculitis (AASV) patients and if this was associated with RXRB genotypes. TNFalpha and IL-10 production were measured in whole blood assay from AASV patients (n = 51) and healthy controls (HC, n = 67). One micromolar of 1,25-(OH)(2) D3, 9-cis RA (9c-RA) or all-trans RA (ATRA) was added to the assay. Genotyping was performed for exons 7 and 2 of the RXRB gene and for a microsatellite in vicinity of the RXRB gene. Lipopolysaccharide (LPS) mediated TNFalpha production and IL-10 were significantly lower in patients. Addition of 1,25-(OH)(2) D3, ATRA or 9c-RA, blunted TNFalpha production, more pronounced in patients. Although all three compounds inhibited IL-10 production significantly in HC, only 1,25-(OH)(2) D3 was found to be effective in patients. Allele distribution of the RXRB microsatellite differed significantly between patients and HC. This was not found for the SNP in exons 2 and 7. Genotype of the latter correlated with the ability of 1,25-(OH)(2) D3 and ATRA to inhibit IL-10 production. We provide immunological evidence for a functional difference in vitamins D and A responsiveness in AASV patients. Since the inhibition of TNFalpha was more effective in patients, vitamin D supplementation might be an additional therapeutical approach. PMID- 19811265 TI - Constant dripping wears away the stone. PMID- 19811266 TI - Waste: an important immune modulator. PMID- 19811267 TI - Apo material as a trigger for inflammation in systemic lupus erythematosus. AB - While several characteristics of systemic lupus erythematosus (SLE) have been investigated, the distinct pathogenetic mechanisms leading to autoimmunity and chronic inflammation are not understood yet. A central role for apo has been implicated in the pathogenesis of SLE and an increased rate of apo or a defective clearance of apo cells have repeatedly been described in SLE patients, which show elevated levels of alpha-interferon (alphaIFN) as well as an enhanced expression of alphaIFN-alpha inducible genes referred to as alphaIFN signature. Recent publications link alphaIFN and apo: apo cell-derived microparticles can directly stimulate plasmacytoid dendritic cells to secret alphaIFN. This review highlights the role of apo material as source for AAg and as a trigger for chronic inflammation in the pathogenesis of SLE. PMID- 19811268 TI - Nph, DC and chromatin-mediated pathogenesis in systemic lupus erythematosus. AB - Nucleosomes (Nuc) are key lupus autoantigen and have been shown to activate several types of immune cells as well as the complement system, resulting in inflammation. The elevated concentrations and the immunostimulatory capacities of circulating Nuc may favour the break of peripheral tolerance in genetically predisposed individuals. Nevertheless, in most cases the signalling pathways involved are not elucidated yet, although Tir8/Sigirr deficiency has been recently shown to enhance cell activation upon exposure to Nuc-containing immune complexes. PMID- 19811269 TI - The interplay between the antiphospholipid syndrome and systemic lupus erythematosus. AB - Antiphospholipid syndrome (APS) was firstly described in systemic lupus erythematosus (SLE), but it was recognized also as a primary APS (PAPS) form. These forms are not always distinguishable, since they show some common clinical/serological manifestations. We actually may deal with: (1) patients initially classified as PAPS gradually developing SLE; (2) patients with SLE and associated APS, whose complications generally affect morbidity and mortality; (3) patients with SLE and positive antiphospholipid antibodies without APS manifestations; the relevant issue in such patients is to provide effective prophylaxis. The close relationship between PAPS and SLE is also supported by: (i) nuclear autoimmunity and (ii) complement activation at least in animal models of APS. Future studies on the genetic background and/or on regulatory suppressive mechanisms may clarify how and why PAPS can evolve into SLE. PMID- 19811270 TI - Role of CCR5 Delta32 bp deletion in RA and SLE. AB - CCR5 and its ligands play important roles in rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). A deletion of 32 bp in its gene leads to the production of a non-functional receptor. Although a protective effect of CCR5 Delta32 for the development of RA has been suggested, future study is required to establish the exact role of this deletion for susceptibility to SLE and lupus nephritis. Also, with regard to the association of CCR5 Delta32 with disease severity in RA and SLE, more data are needed to draw firm conclusions. This might become even more relevant as a CCR5 blocking agent is now available. PMID- 19811271 TI - Total and biologically active CD154 in patients with SLE. AB - CD154, a member of the tumor necrosis factor receptor family, is involved in several biological responses. In the sera of systemic lupus erythematosus (SLE) patients, the levels of sCD154 have been shown to be increased, however, few reports have dealt with the biologically active tetramer. Here, we assessed the biological activity of the serum CD154 tetramer using bioassays for BC activation and production nitrite or peroxide. The patients showed a markedly increased total sCD154 serum concentration (12.5 +/- 8.2 vs. 3.9 +/- 1.2 ng/ml; p < 0.001). ba-sCD154 was significantly increased in non-treated patients (7.4 +/- 3.4 ng/ml, n = 22; p < 0.001) and patients with the highest SLE disease activity index (SLEDAI) scores (5.3 +/- 2.9 ng/ml, n = 8), but not in stable patients (1.3 +/- 1.2 ng/ml, n = 30) whose values were similar to normal healthy donors (NHD; 0.8 +/- 0.2 ng/ml). Patients with SLEDAI above 8 that recovered after successful treatment displayed significantly decreased levels of ba-sCD154. We conclude that the bioassay is a useful tool discriminating active and stable SLE, as well as non-treated patients. PMID- 19811272 TI - Decreased transcription, expression and function of low-density lipoprotein receptor in leukocytes from patients with systemic lupus erythematosus. AB - The low-density lipoprotein receptor (LDLR) is directly involved in the metabolism of low-density lipoprotein (LDL) and its impairment causes accumulation of plasmatic LDL leading to atherosclerosis, a prevalent disease in patients with systemic lupus erythematosus (SLE). We studied LDLR transcription, expression and function in leukocytes patients with SLE and normal healthy donors (NHD). The ratio LDLR/glyceraldehyde-3-phosphate dehydrogenase (GADPH) mRNAs the expression of LDLR and the uptake of LDL-DiI were significantly lower (p < 0.001) in the patients with SLE. On the contrary, patients with SLE had significantly higher (p < 0.0001) levels of total cholesterol, LDL cholesterol and anti-oxLDL autoantibodies (AAb) as compared to NHD. No correlation between LDLR transcription, expression and function with the SLE disease activity index or with treatment was found. The decreased function of LDLR was independent of treatment. It seems dependent on the sterol regulatory binding protein and may be responsible for the increase of plasma LDL cholesterol and oxLDL AAb further increasing the risk of vascular diseases. PMID- 19811273 TI - MoMa from patients with systemic lupus erythematosus show altered adhesive activity. AB - The phagocytosis of apo cells in patients with systemic lupus erythematosus (SLE) has been shown to be impaired. Recognition and engulfment of dying cells are mediated by molecules and receptors that are also involved in cell adhesion. Therefore, we analysed the adhesion capabilities of MoMa from patients with SLE. We employed a colorimetric assay to quantify the adhesion of MoMa to various substrates. In addition, we evaluated the morphological characteristics of the MPhi adhesion cultures by phase contrast microscopy. We have observed various morphologic characteristics depending on the adhesive substrate. The typical spindle shaped MoMa adhered to plastic and a bigger and rounder formed MoMa adhered to immunoglobulin (Ig) coated surfaces. Quantitatively, adhesion to plastic was significantly decreased in some SLE patients. Those patients showing a defective adhesion to plastic simultaneously showed an enhanced adhesion to Ig coated surfaces. These findings suggest that there is an alteration in the integrin and/or FcgammaR mediated stimulation of MoMa from SLE patients and this alteration may be associated with the impaired clearance of apo cells seen in this disease. PMID- 19811274 TI - Serum adipokine levels in patients with systemic lupus erythematosus. AB - In patients with systemic lupus erythematosus (SLE) metabolic alterations are often observed, which may be due to either the disease, the genetic background or the treatment. We studied the serum levels of the adipokines leptin, adiponectin, resistin, visfatin and ghrelin in patients with SLE and controls. Leptin levels were lower and adiponectin, ghrelin and visfatin levels were higher in the patients. No significant differences were encountered for resistin. The values of adipokines were independent of treatment, even after correction for body mass index. Inverse correlations were found among leptin and adiponectin, ghrelin and visfatin. We conclude that adipokines are involved in the metabolic imbalance of patients with SLE. PMID- 19811275 TI - Clearance of apo Nph induces an immunosuppressive response in pro-inflammatory type-1 and anti-inflammatory type-2 MPhi. AB - Nph are crucial for proper host defence. Paradoxically, they also contribute to pathology in various inflammatory diseases. Hence, apo of Nph and subsequent removal from inflamed sites is critical for resolution of inflammation. Apo Nph are recognised and cleared by MPhi, supposedly in a "silent" fashion. MPhi show large heterogeneity, comprising various subsets with different functional and biochemical properties. The contribution of these distinct populations to clearance of apo Nph is as yet unknown. Here, we investigated phagocytosis and subsequent functional responses of in vitro generated pro-inflammatory MPhi1 and anti-inflammatory MPhi2. Although only MPhi2 were capable of efficient Nph phagocytosis, we found that contact with apo Nph excerts immunosuppressive effects on both subsets, skewing them towards an anti-inflammatory state. PMID- 19811276 TI - Altered cell-cell and cell-matrix interactions in the development of systemic autoimmunity. AB - MPhi of mice from the major inbred models of systemic lupus erythematosus (SLE) have an identical defect affecting the activity of the cytoskeletal regulator and G-protein Rho. This abnormality is triggered by apo cells. Strikingly, SLE-prone MPhi show normal Rho activity when cultured in the absence of apo cells. We used gene arrays to identify adhesion-related gene products that are abnormally expressed by MPhi from prediseased 4-6-week-old SLE-prone MRL mice in the presence of serum lipids mimicking apo cells (SL-Apo). MPhi of MRL mice differentially expressed 42 adhesion-related genes in the presence of SL-Apo. Of these, 32 were expressed normally in the absence of SL-Apo. As adhesive interactions play a major role in lymphocyte activation, the detected apo cell dependent abnormality could predispose to the development of autoimmunity. Indeed, several recent genetic studies support a role for adhesion-related genes in the pathogenesis of chronic autoimmunity. PMID- 19811277 TI - C-reactive protein in systemic lupus erythematosus. AB - C-reactive protein (CRP) is an acute phase protein that plays a major role in the regulation of the inflammatory response. It activates the classical complement pathway in a controlled fashion, enhancing the capacity for defence against bacterial infections. It promotes the regulation of MPhi activity through FcgammaR, and is associated with the clearance of apo cells and nuclear antigen, thus becoming a protective molecule against pathogenic autoimmune responses in general, and systemic lupus erythematosus in particular. CRP is also associated with atherosclerosis, both in the general population and in different auto-immune conditions. It plays a double role as a biomarker for vascular risk and as an independent risk factor as it can also perpetuate the inflammatory response. Its multi-task behaviour makes it a pivotal structure both in the comprehension of the pathogenesis of auto-immune and inflammatory responses as well as an important tool in the clinical management of patients. PMID- 19811278 TI - Treatment with DNAse I fosters binding to nec PBMC of CRP. AB - CRP is an important inflammatory marker, however, CRP levels are relatively low in patients with SLE. In addition patients with SLE often display low activities and serum levels for DNase I and complement, respectively. Here we show that DNase I treatment of nec PBMC increased their binding of CRP. Consequently, reduced DNase I activity in patients with SLE may contribute to the impaired opsonisation by CRP of dead cells, exacerbating the clearance defect in SLE of apo and nec cells. PMID- 19811279 TI - oxLDL/beta2GPI complex and anti-oxLDL/beta2GPI in SLE: prevalence and correlates. AB - High levels of oxidized low-density liprotein/beta2 glycoprotein 1 (oxLDL/beta2GPI) complexes and anti-complex IgG as well as IgM have been reported in SLE. We analysed this complex and Ab against the complex in SLE patients and evaluated their relationship with clinical and serological findings, traditional risk factors for atherosclerosis, and subclinical atherosclerosis. The prevalence and the levels of the complex and of anti-complex Ab were significantly higher in systemic lupus erythematosus (SLE) patients than in normal healthy donors (NHD). The titers of oxLDL/beta2GPI were significantly higher in patients with renal involvement and previous thromboembolic episodes and were correlated with the number of risk factors for atherosclerosis, whereas they were significantly lower in patients with neurological involvement. Both IgG and IgM anti-complex Ab were associated with antiphospholipid (APL). In conclusion, the oxLDL/beta2GPI complex as well as Ab against the complex are prevalent in SLE where they seem to be involved in organ damage. PMID- 19811280 TI - T cells demonstrate a Th1-biased response to native beta2-glycoprotein I in a murine model of anti-phospholipid antibody induction. AB - Anti-phospholipid syndrome (APS) is an autoimmune disorder characterized by the presence of autoantibody (AAb) to phospholipid (PL)-binding proteins, such as beta2-glycoprotein I (beta2GPI), and clinical manifestations including thrombosis and/or recurrent pregnancy loss. beta2GPI-reactive T cells are clearly implicated in the generation of these AAb, but the mechanism responsible for their activation remains unclear. We hypothesized that immunization of mice with human beta2GPI, in the context of a potent innate immune activator lipopolysaccharide (LPS), would generate not only high titers of anti-PL AAb, but also a strong beta2GPI-specific T cell response. Healthy, nonautoimmune C57BL/6 mice were immunized repeatedly with human beta2GPI in the presence of LPS. High titers of anti-PL to beta2GPI appeared after the second immunization, with T cell reactivity to beta2GPI detectable only after the fourth immunization. Splenic T cells from these mice proliferated in response to native beta2GPI, alone or bound to anionic PL. These T cells produced IL-2 and IFN-gamma, but not IL-4 or IL-10, indicating a Th1 bias of the beta2GPI-specific response. These findings suggest that T cells responsive to beta2GPI may become activated in APS patients by exposure to their cognate Ag in the context of innate immune activation and a pro inflammatory environment. PMID- 19811281 TI - Elevation of plasma levels of the long pentraxin 3 precedes preeclampsia in pregnant patients with type 1 diabetes. AB - Pregnancy complications, such as preeclampsia (PEc), have an increased incidence among patients with type 1 diabetes (T1DM), possibly because of maternal vascular involvement. The prototypic long pentraxin, pentraxin 3 (PTX3) is an acute phase reactant critically associated with vascular injury. PTX3 concentrations selectively increase in pregnant women with PEc. Here, we measured PTX3 levels in 37 consecutive pregnant patients with T1DM. Compared with PTX3 levels in healthy pregnant women at identical gestational ages, PTX3 was significantly elevated in pregnant women with diabetes. Patients with pre-existing nephropathy, a well characterized microvascular complication of diabetes, have even higher PTX3 concentrations and worse maternal and fetal outcomes. Six/thirty-four diabetic non-nephropatic patients developed PEc: PTX3 levels rose abruptly weeks before PEc manifested (p = 0.0375). PTX3 may represent a valuable marker for early detection and prediction of PEc in patients with T1DM. PMID- 19811282 TI - The translocation of HMGB1 during cell activation and cell death. AB - High-mobility group box protein 1 (HMGB1) is a non-histone nuclear protein with alarmin activity. When present in an extracellular location, HMGB1 can activate the innate immune system and promote inflammation in conditions such as sepsis. To exert these activities, HMGB1 must transit from the nucleus, through the cytoplasm, to the outside of the cell. This process can occur during cell activation as well as cell death. In murine macrophages (MPhi), stimulation of TLR3 and TLR4, but not TLR9, can cause HMGB1 translocation. With cell death, necrosis can lead to extracellular HMGB1 by a passive mechanism. With apoptosis, HMGB1 is only released during secondary necrosis, when cell permeability barriers break down. Since agents that stimulate MPhi can also induce apoptosis, HMGB1 release following TLR stimulation may also reflect a contribution from dead cells, suggesting a common mechanism for protein release in activation and death. PMID- 19811283 TI - AGE and their receptor RAGE in systemic autoimmune diseases: an inflammation propagating factor contributing to accelerated atherosclerosis. AB - Systemic autoimmune diseases are associated with inflammation, and oxidative stress favouring the formation of advanced glycation endproducts (AGE), able to modulate cellular functions by activation of receptor for advanced glycation endproducts (RAGE). As RAGE expression is increased in an inflammatory milieu, present in patients with systemic autoimmune diseases, these patients are especially prone for the deleterious effects of AGE. Interaction of AGE with RAGE leads to intracellular signalling, and subsequent expression of adhesion molecules, chemokines, pro-inflammatory cytokines and up-regulation of RAGE itself. The AGE-RAGE interaction might act as a pro-inflammatory loop in these patients, contributing to chronic low grade inflammation rendering these individuals susceptible for development of accelerated atherosclerosis. PMID- 19811284 TI - Oxidation of the alarmin high-mobility group box 1 protein (HMGB1) during apoptosis. AB - The architectural chromosomal protein high-mobility group box 1 protein (HMGB1) acts as an alarmin when released from cells. It is involved in the pathogenesis of inflammatory and autoimmune diseases. HMGB1 can undergo post-translational modifications including oxidation. However, the mechanisms and functional relevance of HMGB1 oxidation are not yet understood. Increased concentrations of reactive oxygen species (ROS) have been reported during apoptosis and necrosis. Hence, we investigated the oxidative status of HMGB1 in dead cells. Immunoblot analyses under reducing and non-reducing conditions revealed that HMGB1 is oxidized in dead cells. Moreover, tagging of oxidized cysteine residues by a maleimide moiety linked to polyethylene glycol showed that HMGB1 passively released from primary and secondary necrotic cells was predominantly oxidized. Also HMGB1 in plasma of patients with systemic lupus was reversibly oxidized. In conclusion, HMGB1 undergoes reversible oxidative modifications at cysteine residues during cell death, which may modulate its biological properties. PMID- 19811285 TI - High mobility group B2 is secreted by myeloid cells and has mitogenic and chemoattractant activities similar to high mobility group B1. AB - High mobility group B box (HMGB) proteins are a family of chromatin proteins made up of two basic DNA binding domains, HMG box A and B, and a C-terminal acidic tail. HMGB have a highly conserved sequence, but different expression pattern: HMGB1 is almost ubiquitous, whereas the others are highly expressed in only a few tissues in adults. We previously demonstrated that HMGB1 is released by necrotic cells and has chemoattractant activity for inflammatory and stem cells, via binding to receptor for advanced glycation endproducts (RAGE). HMGB1 can be actively secreted by inflammatory cells. Here, we report that also HMGB2 can be secreted by THP-1 cells, and promotes proliferation and migration of endothelial cells. These functions of HMGB2 are exerted via engagement of RAGE, whose blockade completely abrogates cell responses. Since extracellular HMGB2 has been detected in the blood and other biological fluids, it might be necessary to target HMGB2 at the same time as HMGB1 for therapeutical efficacy. PMID- 19811286 TI - Hyperthermia in combination with X-irradiation induces inflammatory forms of cell death. AB - Autoimmune diseases and cancer can be treated by influencing the immune system. Apo and nec cells are strong modulators of the immune system contributing to anti inflammatory and pro-inflammatory responses, respectively. We examined which form of cell death was induced by HT and X-irradiation. Nec was the prominent form of cell death after combined treatment and the amount of dead cells was higher when exposing the cells to radiation before HT. Combined applications further led to an increased percentage of cells in a more radioresponsive G2 cell cycle phase. The danger signal HMGB1 is released when combining HT with radiation, a further hint that those treatments may induce inflammation and immune activation. We conclude that immune responses are appropriately adapted to the damage that has occurred and may contribute to anti-cancer immunity or chronic autoimmunity, respectively. PMID- 19811287 TI - Sodium and potassium urate crystals differ in their inflammatory potential. AB - Uric acid (UA) is identified as a danger signal released from dying cells. It precipitates in sodium-rich extracellular fluid and in potassium-rich intracellular fluid as monosodium urate (MSU) and monopotassium urate (MPU), respectively. Here, we examined the structural and functional features of these crystals. In contrast to MPU MSU crystals induced reactive oxygen species production and release of pro-inflammatory cytokines in whole blood ex vivo assays. These results show that the cation of urate crystals determines the response of innate immune cells, indicating that the micromilieu at the site of crystal formation is important for their inflammatory potential. PMID- 19811288 TI - Apoptotic and necrotic cells as sentinels of local tissue stress and inflammation: response pathways initiated in nearby viable cells. AB - Virtually all cells in the body have the capacity to recognize and respond to dead cells. Viable cells discriminate apo from nec targets via distinct cell surface receptors. Engagement of these receptors induces "recognition-dependent" signaling events in viable responding cells that differ for apo vs. nec targets. Although "engulfment-dependent" signaling events also contribute to the response by viable cells, these events do not differ for apo vs. nec targets. While many signaling events are conserved across diverse cell lineages, other signaling events, especially those involving Akt, demonstrate lineage-specific variation. Whereas apo targets activate Akt in MPhi, they inhibit Akt in kidney epithelial cells. Differences in the responses to dead targets by viable migratory cells, such as MPhi, and viable fixed cells, such as kidney epithelial cells, permit cell-specific adaptations to local environmental change or stress. We propose that dead cells (apo and nec) act as sentinels to alert nearby viable cells to local environmental change or stress. PMID- 19811289 TI - Apototic cell-derived membrane vesicles induce CD83 expression on human mdDC. AB - A characteristic of apo cell death is the shedding of membrane vesicles from the dying cell. This process is also referred to as apo membrane blebbing. These apo particles contain various autoantigens and are effectively engulfed by phagocytes. A defective phagocytosis has been described in autoimmune diseases like systemic lupus erythematosus and this defect might lead to an accumulation of apo cells and bodies. Thus, we investigated the interactions between apototic cell-derived membrane vesicles (ACdMV), and myeloid dendritic cell (DC). ACdMV were isolated from the supernatant of apo lymphocytes. These isolated ACdMV were morphologically characterized as membrane coated vesicles of an average size of 500 nm. Coincubating isolated ACdMV with iDC we observed CD83 surface expression of the latter. Accumulation of ACdMV may contribute to an inflammatory immune response in autoimmune diseases. PMID- 19811290 TI - Both early and late apoptotic blebs are taken up by DC and induce IL-6 production. AB - During apo blebs, containing nuclear components, are formed at the cells' surfaces. When these blebs separate from the dying cell an apo cell body remains. The contents of apo blebs are modified and can be released, especially in patients with systemic lupus erythematosus (SLE) since impaired clearance of apo material has been observed in this autoimmune condition. Accordingly, autoantibodies present in subjects with SLE bind to apo blebs. Based on AnxA5 binding, and permeability for PI, we show that apo blebs can be categorized as early (AnxA5(+)/PI(- )) or late (AnxA5(+)/PI(+)) apo ones. Both forms of blebs contain apo-induced chromatin modifications and are efficiently phagocytosed by dendritic cell (DC). Uptake by DC of late, but also early apo blebs, stimulate DC to produce IL-6. This bleb-induced effect on DC may be an important step in the initiation of the autoimmune responses in SLE. PMID- 19811291 TI - Cells induced to undergo apo in the presence of the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone stimulate Mo derived phagocytes to secrete proinflammatory cytokines. AB - In contrast to nec, the apo is not accompanied by local inflammation. The immunosuppressive effects of apo cells have been repeatedly reported and a dysregulation of apo is discussed to play a major role in the pathogenesis of autoimmune disorders. The intracellular executioners of apo are the cysteine aspartic acid proteases, also known as caspases that cleave a variety of intracellular substrates and mediate the morphological changes observed during apo. The association of autoimmune diseases with defects in caspase function indicates the necessity for functional integrity of caspases in the apo cell death machinery. Here, we describe that cells undergoing apo in presence of the pancaspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone stimulated MPhi to secrete proinflammatory cytokines. These findings indicate that caspase signalling is of central importance for silent and non- or anti-inflammatory cell death. PMID- 19811292 TI - Lipoxin A4 receptor dependent leishmania infection. AB - The lipoxin A4 receptor (ALX) is an important target of LxA4 in synovial tissues of patients with inflammatory arthritis. Previously this receptor was known as the FPRL-1 on PMN and shown to interact with acute phase proteins and a variety of peptides. ALX signalling can either activate or deactivate PMN functions. In this study, we found that both LxA4 and a chemotactic lipid leishmania chemotactic factor released by the parasite leishmania increased infectivity of this pathogen in an ALX dependent fashion. This functional characterization of ALX could lead to development of novel, therapeutic targets for treatment inflammatory diseases. PMID- 19811293 TI - Apoptosis-related changes in plasma membrane glycoconjugates of peripheral blood lymphocytes in rheumatoid arthritis. AB - Control of apoptosis (apo) is very important for diagnosis, prognosis and treatment of rheumatoid arthritis (RA). Recently, we found that an appearance of specific cell surface GC is attributable to apo and developed lectin-induced agglutination test for apo evaluation. The aim of current study was to estimate changes in surface GC expression in peripheral blood lymphocytes (PBL) of normal healthy donors (NHD) and patients with RA, measured by cell agglutination with the galactose-specific VAA lectin and by lectin-cytochemical analysis. In parallel, these changes in apo incidence were evaluated by the detection of cells with sub-G1 DNA content. The data revealed an increased level of apo in lymphocytes of RA patients (n = 29), and increased cell surface GC expression in lymphocytes of NHD (n = 18). A correlation (R = 0.708) was observed between these two indicators. Specific changes in cell surface GC can be effectively used for the detection of apo cells in RA and other autoimmune disorders. PMID- 19811294 TI - Radio-immunological mechanisms of anti-inflammatory treatment: is there a way from the past into the future? AB - The anti-inflammatory efficiency of low-dose radiotherapy (LD-RT) for degenerative joint disorders was demonstrated over decades but had no explanation on a cellular or molecular level. As inflammatory diseases are the results of complex and pathologically unbalanced cellular and molecular interactions more recent in-vivo and in-vitro data will be discussed for possible explanation of the mechanism underlying ant-inflammatory LD-RT.2. PMID- 19811295 TI - Activation-induced cell death and total Akt content of granulocytes show a biphasic course after low-dose radiation. AB - Low-dose radiation (single doses from 0.3 to 1.0 Gy) in clinical practice is mostly used to treat patients with several inflammatory diseases and painful degenerative disorders. Low-dose radiation is known to exert anti-inflammatory effects. However, the molecular and cellular mechanism are not fully analysed and most of the observed effects are based on empirical studies. We investigated the effects of low-dose radiation on the activation-induced cell death of polymorph nuclear granulocytes (PMN). A biphasic appearance of cell death in irradiated PMN was observed, displaying a relative maximum at 0.3 Gy and minimum at 0.5 Gy, respectively. This biphasic course of cell death was coincident with the protein level of total cellular Akt. We conclude that low-dose radiation exerts immunomodulatory effects on PMN contributing to the observed anti-inflammatory effects in clinical applications. PMID- 19811296 TI - Activator protein 1 shows a biphasic induction and transcriptional activity after low dose X-irradiation in EA.hy.926 endothelial cells. AB - Low dose radiotherapy (LD-RT) is known to exert an anti-inflammatory effect. The underlying molecular mechanisms, however, are still a matter of actual research. We have recently shown that LD-RT of stimulated EA.hy.926 endothelial cells (EC) resulted in a biphasic DNA-binding and transcriptional activity of NF-kappaB in parallel with a biphasic course of leukocyte adhesion. Here we report, that following low dose X-irradiation, an increased activator protein 1 (AP-1) DNA binding activity was observed in EC with a first relative maximum at 0.3 Gy as analysed by electrophoretic mobility shift assay. AP-1 activity then decreased at doses between 0.5 and 1 Gy and subsequently increased again at 3 Gy. This biphasic profile was confirmed on the transcriptional level by an AP-1 specific chemoluminescence reporter assay. In conclusion, the discontinuous dose response of AP-1 activation may add a further facet to the plethora of mechanisms contributing to the anti-inflammatory efficacy of LD-RT. PMID- 19811297 TI - Whole body low dose irradiation improves the course of beginning polyarthritis in human TNF-transgenic mice. AB - Rheumatoid arthritis (RA) displays a chronic inflammatory joint disease, accompanied by symmetric polyarthritis (PA) which evokes synovial inflammation, cartilage damage, and bone erosion. Patients with RA are routinely treated by immunosuppressive drugs. The therapy of inflammatory diseases and degenerative disorders with Low-dose radiotherapy (LD-RT) (single doses from 0.3 to 1.0 Gy) represents a low cost therapy with low toxicity, and is able to substitute at least in part treatment with drugs. The efficiency of LD-RT has already been proven in several animal models of inducible arthritis. In the present study we used a human TNF transgenic mouse model to examine the effects of LD-RT on PA. We observed a significant temporal improvement of the clinical progression of disease when mice were irradiated at the beginning of the disease. These data emphasize the role of LD-RT in clinical settings to treat patients with chronic and degenerative disorders and diseases. PMID- 19811298 TI - Endogenous type I interferon inducers in autoimmune diseases. AB - Type I interferon (IFN) is produced by the innate immune system in several autoimmune diseases, such as systemic lupus erythematosus (SLE), polymyositis, and systemic sclerosis. In these diseases, immune complex (IC)-containing DNA or RNA may act as endogenous IFN inducers. The abilities of these IC to reach the endosomes in the plasmacytoid dendritic cells (PDC) cause the intracellular toll like receptor (TLR) to initiate a cascade of transcription factors--a critical step in triggering type I IFN production. A special configuration of the nucleic acid (NA), such as CpG-rich non-methylated DNA or GU-rich RNA, appears crucial. However, other components of the IC, like HMGB1, may also be necessary. Studies regarding the genetic background of autoimmune diseases suggest that variants of genes involved in both IFN production and response are associated with disease susceptibility. This knowledge is important for the development of new therapeutic strategies in autoimmune diseases. PMID- 19811299 TI - PDC expressing CD36, CD61 and IL-10 may contribute to propagation of immune tolerance. AB - Human plasmacytoid dendritic cells (PDC) are blood dendritic cell antigen 2 (BDCA2) and blood dendritic cell antigen 4 (BDCA4) positive leukocytes that do not express common lineage markers. They have been described as proinflammatory innate immune cells and are the major source of alphaIFN in the human body. PDC derived secretion of type I IFNs upon triggering of nucleic acid-sensing toll like receptors (TLR) primes immune cells to rapidly respond to microbial stimuli and promotes a Th1 response. Here, we report that human PDC express CD36 and CD61 (beta3 integrin), both involved in uptake of apoptotic cells and in induction of tolerance. Freshly isolated PDC and PDC within human blood leukocytes constitutively express IL-10. Thus, PDC may possess a so far neglected role in propagation of immune tolerance. PMID- 19811300 TI - Relationship between serum levels of TGF-beta1 and clinical parameters in patients with rheumatoid arthritis and Sjogren's syndrome secondary to rheumatoid arthritis. AB - TGF-beta1 is a pleiotropic cytokine, which prevents inappropriate autoimmune responses and balances the requirements of proper immune cell levels during pathologic states that trigger the immune response. We assessed the serum levels of TGF-beta1 and determined the relationship between TGF-beta1 and clinical parameters in patients with rheumatoid arthritis (RA) and Sjogren's syndrome (SS) secondary to RA (SS + RA). Comparison of the serum levels of TGF-beta1 in patients with RA, SS + RA and NHD differed significantly (51.7 +/- 12.4 ng/ml (RA); 33.0 +/- 3.1 ng/ml (SS + RA) and versus 31.6 +/- 2.0 ng/ml (NHD)). We further found correlations between TGF-beta1 levels and radiologically defined joint damage determined by the Steinbrocker scoring system, symptoms and signs of SS. We conclude that serum levels of TGF-beta1 may reflect ongoing autoimmune inflammation and correlate with joint damage in RA. PMID- 19811301 TI - DNA-hydrolyzing Ab: is catalytic activity a clue for physiological significance? AB - Ab with DNA-hydrolyzing properties were described in autoimmune pathologies, such as SLE and RA, and in other autoimmune diseases, including chronic lymphocytic leukemia, AIDS, and others. The disease-associated DNA-binding AAb penetrate cell membrane and enter the nucleus. Intracellular entry of anti-DNA was linked to cellular damage and apo. Here we discuss the possible pathological process induced by DNA-cleaving Ab in the nucleus, where these Ab may induce apo. Apo processes, e.g., induced by DNA-hydrolyzing Ab, may underlie a number of syndromes observed in autoimmunity thus providing the grounds for the pathological role of DNA-hydrolyzing Ab in these diseases. PMID- 19811302 TI - Suppression of ongoing experimental allergic encephalomyelitis in DA rats by novel peptide drug, structural part of human myelin basic protein 46-62. AB - Previously, we demonstrated that autoantibodies (AAb) in multiple sclerosis (MS) reveal site-specific binding and cleavage toward myelin basic protein (MBP) epitope library. We have found several fragments of MBP immunodominant in terms of AAb binding. Here, we applied these peptides to DA rats with induced protracted relapsing experimental allergic encephalomyelitis (EAE) most closely related to MS. DA rats with EAE induced by syngenic spinal cord homogenate in complete Freund's adjuvant were treated by nasal route with human MBP 46-62, 81 102, 124-139, 147-170, and Copaxone. MBP 124-139 and 147-170 displayed only mild therapeutic effects but MBP 46-62 significantly reduced EAE, reflected by lower clinical scores and shorter EAE duration compared to controls. PMID- 19811303 TI - Protein destabilizing agents induce polyreactivity and enhanced immunomodulatory activity in IVIg preparations. AB - Normal pooled human IVIg are produced using various blood protein fractionation technologies and as a result they may well differ in their biological properties. We have demonstrated that exposure of IVIg, for a period as short as 15 min, to protein-destabilizing agents like acidic pH, ROS or pro-oxidative ferrous ions dramatically increases the panel of recognized Ag including pro-inflammatory cytokines. We now show that exposure of IVIg to ferrous ions modifies some IgG molecules without denaturating them and enhances the protective activity of the preparation in experimental septic shock. PMID- 19811304 TI - Imaging inflammation in real time--future of nanoparticles. AB - The detection of subclinical early inflammation in autoimmune diseases is an important but currently technically demanding approach to direct initial diagnosis and subsequent choice of therapy. Recent advances in imaging using NP provides the potential to detect cellular recruitment, vascular activation or leakage at a subclinically stage of disease and may provide predictive "biomarkers" of future pathogenesis. The NP used are either untargeted and taken up by phagocytic cells, or are linked to a ligand, targeting localisation to the site of inflammation. Techniques, varying from MRI and fluorescence to Raman spectroscopy are being employed. In this short review, we summarise many of the recent developments in the field of NP imaging related to inflammation. PMID- 19811305 TI - Do B cells play a role in the pathogenesis of juvenile idiopathic arthritis? AB - Juvenile idiopathic arthritis (JIA) is the most common cause of chronic arthritis in childhood and adolescents and encompasses a heterogeneous group of diseases. The role of B cells (BC) in autoimmune diseases has been put in a new perspective due to the promising results of BC depleting therapies in RA. Experiments in mouse models have shed new light on the Ab-independent role of BC in the pathogenesis of autoimmune diseases. We discuss whether BC play a role in the pathogenesis of JIA appraising the question for an immunological basis of BC directed therapy. PMID- 19811306 TI - Pathogenesis of Churg-Strauss syndrome: recent insights. AB - Churg-Strauss syndrome (CSS) is a rare systemic necrotizing vasculitis associated with granuloma formation and severe blood and tissue eosinophilia. CSS occurs almost exclusively in patients with asthma. Its pathogenesis remains largely unknown, as triggering factors for CSS development have not been identified so far. AAb, such as anti-neutrophil cytoplasmic autoantibodies, are found in less than half of patients and possibly constitute a subtype of CSS with different clinical behaviour. On a cellular level, CSS is characterized by a strong Th2 type immune response. Th2-associated cytokines such as IL-4, IL-13 and IL-5 may precipitate the severe eosinophilia in CSS, while migration of Eos to inflammatory sites is possibly mediated by eotaxin-3. This review summarizes recent advances in the knowledge on epidemiology, clinical features, and pathogenesis of CSS. PMID- 19811307 TI - Lentiviral gene transfer of CTLA4 generates B cells with reduced costimulatory properties. AB - Peripheral T-cell (TC) tolerance can be induced by tolerogenic antigen-presenting cell (APC). A prerequisite is the reduction or blockade of B7 of APC. Besides dendritic cell, B cells can be used as APC. Here, we show the generation B cells with reduced B7 expression by lentiviral transduction of endoplasmic reticulum (ER)-directed CTLA4. Vectors coding for the human CTL4-Ig were used for the human B-cell line Raji. Transduction efficiency was over 90% (MOI = 3). For the murine B-cell line A20 and for primary mouse B cells, murine CTLA4 was used. We show that B cells with reduced B7 expression reduce the antigen (Ag) specific TC proliferation in vitro. B cells expressing an ER-directed CTLA4 may reduce Ag specific immune responses. PMID- 19811308 TI - The 12/15-lipoxygenase pathway promotes osteoclast development and differentiation. AB - Although 12/15-lipoxygenase (12/15-LO) has been implicated as negative regulator of systemic bone mass in mice and humans, the underlying mechanisms remain elusive. Here, we show that 12/15-LO is a positive regulator of osteoclast (OC) development. Enzymatic inhibition as well as genetic ablation of 12/15-LO significantly impaired osteoclastogenesis. Conversely, addition of the 12/15-LO derived eicosanoids 12- and 15-HETE augmented differentiation of precursors into fully matured OCs. Together these data point towards a crucial role of 12/15-LO in the regulation of OC development. Therefore, 12/15-LO and its human homologue 15-LO may display novel targets for the treatment of diseases such as osteoporosis and rheumatoid arthritis. PMID- 19811309 TI - Leukocyte and platelet activation in patients with giant cell arteritis and polymyalgia rheumatica: a clue to thromboembolic risks? AB - Ischemia is a leading causes of morbidity in giant cell arteritis (GCA). We studied circulating platelets and leukocytes in patients with GCA and with polymyalgia rheumatica. Normal healthy donors (>60 a) served as controls. Patients had a significantly greater fraction of platelets expressing P-selectin, of platelet-Nph and platelet-Mo aggregates, and of Nph and Mo expressing tissue factor. These differences were correlated with the percentage of platelets expressing P-selectin and were not influenced by clinical features or by systemic inflammation. Activated circulating leukocytes and platelets could contribute to indolent vessel inflammation and possibly to thromboembolic events in patients with systemic large vessel vasculitis. PMID- 19811310 TI - HLA-DQA1 gene expression profiling in oligoarticular JIA. AB - Polymorphisms in the upstream regulatory region of the HLA class II DQA1 gene are currently defined by 10 different alleles. Two of them carrying a Y-box mutation are associated with susceptibility to oligoarticular juvenile idiopathic arthritis (OA-JIA). We investigated allele-dependent differences in HLA-DQA1 gene expression in OA-JIA patients. In cells from affected joints compared to peripheral blood, gene expression of HLA-DRA as well as total HLA-DQA1 was significantly upregulated. Differential analyses of HLA-DQA1 allelic expression showed DQA1*02 and *04 to be comparatively increased. Intra-articular upregulation of HLA-DQA1 was predominantly observed for the OA-JIA associated allele HLA-DQA1*04. Nevertheless, the Y-box mutation of the disease-associated allele DQA1*0401 was not a common denominator for expression behaviour. PMID- 19811311 TI - In this issue: on miRNAs connecting cancer with inflammation and novel concepts in immunology and vaccine design. PMID- 19811312 TI - miR-155: on the crosstalk between inflammation and cancer. AB - MicroRNAs are short non-coding RNAs that posttranscriptionally modulate the expression of multiple target genes and are thus implicated in a wide array of cellular and developmental processes. miR-155 is processed from BIC, a non-coding transcript highly expressed in both activated B and T cells and in monocytes/macrophages. miR-155 levels change dynamically during both hematopoietic lineage differentiation and the course of the immune response. Different mouse models developed recently indicate that miR-155 plays a critical role during hematopoiesis and regulates lymphocyte homeostasis and tolerance. A moderate increase of miR-155 levels is observed in many types of malignancies of B cell or myeloid origin, and transgenic over-expression of miR-155 in mice results in cancer. While the high levels of miR-155 reached transiently during the course of the immune response remain unharmful for the organism, the reason why a moderate up-regulation of miR-155 can lead to cancer remains obscure. As prolonged exposure to inflammation can lead to cancer, the permanent up regulation of miR-155 might be a link between the two. Therefore, designing miR 155 based therapies will require a better understanding of the molecular basis of its action as well as of how miR-155 levels are regulated in a cell-specific manner. PMID- 19811313 TI - HIV-1 Tat-based vaccines: an overview and perspectives in the field of HIV/AIDS vaccine development. AB - The HIV epidemic continues to represent one of the major problems worldwide, particularly in the Asia and Sub-Saharan regions of the world, with social and economical devastating effects. Although antiretroviral drugs have had a dramatically beneficial impact on HIV-infected individuals that have access to treatment, it has had a negligible impact on the global epidemic. Hence, the inexorable spreading of the HIV pandemic and the increasing deaths from AIDS, especially in developing countries, underscore the urgency for an effective vaccine against HIV/AIDS. However, the generation of such a vaccine has turned out to be extremely challenging. Here we provide an overview on the rationale for the use of non-structural HIV proteins, such as the Tat protein, alone or in combination with other HIV early and late structural HIV antigens, as novel, promising preventative and therapeutic HIV/AIDS vaccine strategies. PMID- 19811314 TI - Neutropenia and primary immunodeficiency diseases. AB - Primary immunodeficiency diseases (PID) are a heterogeneous group of congenital disorders of the immune system leading to recurrent infections, autoimmunity, malignancies, and hematological disorders. This review focuses specifically on inherited disorders associated with neutropenia, which may occur in isolation or as a feature of more complex immune disorders. It has been known for a long time that defined immunodeficiency syndromes, such as CD40L deficiency, WHIM syndrome, or Chediak Higashi syndrome, may be associated with neutropenia even though the mechanisms are poorly understood. In some PID, neutropenia may result from chronic viral infection or from autoimmunity. Recently, the identification of several novel genetic defects (e.g., p14-deficiency, HAX1-deficiency, AK2 deficiency) has shed light on the pathophysiology of congenital neutropenia. This review summarizes the clinical, immunological, and genetic features of congenital neutropenia syndromes. PMID- 19811315 TI - Can proliferation signal inhibitor-induced Tregs really reflect transplantation tolerance in clinical solid organ transplantation? AB - Exploring new immunosuppressive strategies inducing donor-specific hyporesponsiveness is a great challenge in transplantation. For this purpose, monitoring the alloimmune response is a critical step to carrying out protolerogenic immunosuppressive protocols. Regulatory T cells (Tregs), known as controlling various immune responses, were found to play an important part in allograft transplant tolerance. It is said that Rapamycin (RAPA), one of the proliferation signal inhibitors (PSIs), could achieve true tolerance through the induction of Tregs in clinical trials. Can PSI-induced Tregs in peripheral blood mononuclear cells (PBMC) really reflect transplantation tolerance in clinical solid organ transplantation? PMID- 19811316 TI - Molecular mechanisms of viral immune evasion proteins to inhibit MHC class I antigen processing and presentation. AB - Viral products inhibit MHC class I antigen processing and presentation via three major pathways: inhibition of major histocompatibility complex (MHC) class I expression on cells, blockade of peptide trafficking and loading on MHC class I molecules, and inhibition of peptide generation in host cells. Viral products also interfere with IFN-gamma -mediated JAK/STAT signal transduction in cells. These results imply that viral proteins probably inhibit the function of IFN gamma in MHC class I antigen presentation via inactivation of JAK/STAT signal transduction in host cells. Mechanisms of viral products to inhibit IFN-gamma mediated MHC class I antigen presentation were summarized in this literature review. PMID- 19811317 TI - In this issue: new topics in miRNA and transplantation immunology. PMID- 19811318 TI - Logic and extent of miRNA-mediated control of autoimmune gene expression. AB - Over the past few decades, multiple mechanisms have emerged that operate to prune the lymphocyte repertoire of self-reactive specificities and maintain immunological tolerance. Multiple families of small noncoding RNAs known as microRNAs (miRNAs) target immune transcripts to fine-tune gene expression and turn on negative feedback loops. Both of these actions are crucial to limit co stimulation, set precise cellular activation thresholds, curtail inflammation, control lymphocyte growth, and maintain regulatory T cell homeostasis and suppressive function. Analysis of predicted miRNA-mediated regulation of 72 lupus susceptibility genes in humans and mice reveals most contain numerous target sites for over 140 miRNAs conserved in mammals. MECP2, ROQUIN/RC3H1, BCL2, BIM, and PTEN contain over 50 miRNA target sites each, highlighting the need to control their final protein products with enormous precision to maintain the balance between immunity and tolerance. Overlap among targets of individual miRNAs is considerable, with each miRNA targeting a median of nine autoimmune genes. Three miRNAs--miR-181, miR-186, and miR-590-3p--together are predicted to target over 50% of all lupus genes. Also, a single miRNA cluster located at 14q32.31 containing 11 miRNAs is predicted to regulate 48 lupus susceptibility genes. Dysregulation of single or a few miRNAs or miRNA clusters can result from genetic variation, hormonal influences, or environmental triggers including EBV infection. In the light of this vast and promiscuous miRNA-mediated regulation of autoimmune genes it is anticipated that changes in miRNA levels or their target sequences will help explain susceptibility to complex autoimmune diseases. PMID- 19811319 TI - MicroRNAs and Epithelial Immunity. AB - MicroRNAs are required for development and maintenance of the epithelial barrier. It is hypothesized that microRNAs are involved in regulating epithelial anti microbial defenses by targeting key epithelial effector molecules and/or influencing intracellular signaling pathways. Additionally, aberrant microRNA expression has been implicated in the pathogenesis of various diseases at the skin and mucosa. Increased understanding of the role of microRNAs in epithelial immunoregulation and identification of microRNAs of pathogenetic significance will enhance our understanding of epithelial immunobiology and immunopathology. PMID- 19811320 TI - Clinical relevance of immune monitoring in solid organ transplantation. AB - Transplant recipients receive immunosuppressive drugs to prevent graft rejection and graft loss. Immunosuppressive drugs are dosed according to empiric treatment protocols, which consider the risk of under- or oversuppression, rejection, infection, cancer, and drug side effects. An individualized immunosuppressive protocol based on immune monitoring would be useful for avoiding undesired effects. In retrospective studies, numerous immune parameters were shown to be associated with clinical events. Their suitability for clinical application has to be proven in prospective studies. A multiparametric immune monitoring program, performed frequently and lifelong, might be the appropriate approach to predict dysfunction of the graft before clinical symptoms occur. PMID- 19811321 TI - Role of IDO in organ transplantation: promises and difficulties. AB - Induction of donor-antigen-specific immunological tolerance still remains the "holy grail" in organ transplantation. Recently, Indoleamine-2,3 Dioxygenase (IDO)--a tryptophan degrading enzyme--has been shown to be implicated in one of nature's most impressive examples of tolerance, which is maternal acceptance of the semi-allogeneic foetus. Although many experimental findings propose IDO as a key player in induction and maintenance of peripheral tolerance, scepticism exists as to whether IDO represents a promising therapeutic target with clinical relevance. In this review article we will discuss the role of IDO in transplantation and take a critical look at IDO-based therapeutic strategies. PMID- 19811322 TI - Innate immune signaling pathways in animals: beyond reductionism. AB - The immune system plays a crucial role in the maintenance of the stability and equilibrium of the internal environment in living organisms. The field of animal innate immunity has been the global focus of immunological research for decades. It is now known that the functions of innate immunity inevitably rely on the action of the molecular machines of the cascades or network of immune signaling pathways. Up to date, many researches on the immune signaling pathways in animals were focused on identifying the component functions or cascade molecules in details, which essentially followed a reductionist paradigm without paying high attention to the integrated features. The main purpose of this article was dedicated to accentuating the shift of this field from a reductionist to a systemic view. First, the former part of this article made efforts to summarize the main aspects of the signaling pathways of animal innate immunity including the web resources, the recapitulation of highlighted pathways, the cross-talks, and the evolutionary considerations, which heavily emphasized the integrated characteristics of the immune signaling pathways. Subsequently, the later part of this article was based on the holistic feature of the immune signaling pathways, mainly dedicated to propose a novel hypothesis. From a whole perspective, the oscillating balance hypothesis was deliberately formulated to characterize the holistic pattern of the signaling transduction network of animal innate immune system, which might help to understand some immunological phenomena through the integral principle of the immune network. PMID- 19811323 TI - Molecular mechanisms of IFN-gamma to up-regulate MHC class I antigen processing and presentation. AB - IFN-gamma up-regulates MHC class I expression and antigen processing and presentation on cells, since IFN-gamma can induce multiple gene expressions that are related to MHC class I antigen processing and presentation. MHC class I antigen presentation-associated gene expression is initiated by IRF-1. IRF-1 expression is initiated by phosphorylated STAT1. IFN-gamma binds to IFN receptors, and then activates JAK1/JAK2/STAT1 signal transduction via phosphorylation of JAK and STAT1 in cells. IFN-gamma up-regulates MHC class I antigen presentation via activation of JAK/STAT1 signal transduction pathway. Mechanisms of IFN-gamma to enhance MHC class I antigen processing and presentation were summarized in this literature review. PMID- 19811324 TI - Bendamustine in chronic lymphocytic leukemia: the future is combination therapy. PMID- 19811325 TI - Molecular determinants of prognosis in acute myeloid leukemia (AML) with normal karyotype. PMID- 19811326 TI - Haploidentical hematopoietic stem-cell transplantation for non-Hodgkin lymphoma with bone marrow involvement. AB - Here, we report the preliminary results of haploidentical hematopoietic stem cell transplantation (Haplo-HSCT) with granulocyte-colony-stimulating factor (G-CSF) mobilized bone marrow grafts without T-cell depletion for 10 patients with refractory non-Hodgkin lymphoma accompanied by bone marrow involvement. Eight patients received a conditioning regimen consisting of high-doses of cytarabine and cyclophosphamide with total body irradiation, whereas two cases were preconditioned with busulfan, thiotepa, and cyclophosphamide. All patients had rapid hematopoietic engraftment with the mean time for neutrophil and platelet recovery being 16.6 days and 19.2 days, respectively. Three cases died within 6 months after transplantation from severe acute graft-versus-host disease, fungal infection, or relapse. The others are currently alive in complete remission at a median follow-up of 60.71 months (range: 44-81 months). The results here suggest that haplo-HSCT might provide an opportunity of myeloablative therapy for refractory lymphoma with marrow infiltration. PMID- 19811328 TI - Diet and non-Hodgkin lymphoma risk--food for thought. PMID- 19811329 TI - Autoimmune cytopenia in chronic lymphocytic leukemia/small lymphocytic lymphoma: changes in clinical presentation and prognosis. AB - Improved medical care could have altered the clinical presentation and survival of patients with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) complicated by autoimmune disease cytopenia (AID cytopenia). We reviewed the clinical characteristics, treatment, and outcome of AID cytopenia that was diagnosed in 75 (4.3%) of 1750 patients with CLL seen at a single institution over 10 years. When compared with the historical reported data, our study shows a lower rate of autoimmune hemolytic anemia (2.3%), and similar rates of immune thrombocytopenia (2.0%), and pure red blood cell aplasia (0.5%). AID cytopenia occurred at all stages of CLL, responded well to treatment, did not alter overall survival, and contributed to death in only 6 (12%) patients. We propose that more sensitive and accurate diagnostic methods for CLL have decreased the perceived prevalence of AID cytopenia and that improvements in management could have increased the survival of these patients. PMID- 19811330 TI - Dietary patterns and the risk of non-Hodgkin lymphoma: the Multiethnic Cohort. AB - We examined dietary patterns and the risk of non-Hodgkin lymphoma (NHL) in the Multiethnic Cohort, which includes more than 215,000 Caucasians, African Americans, Japanese-Americans, Native Hawaiians, and Latinos aged 45-75 at baseline. All subjects completed a validated food-frequency questionnaire. After a median follow-up time of 10 years, we identified 939 incident NHL cases through linkages with tumor registries. Three patterns, "Vegetables,""Fruit and Milk," and "Fat and Meat," were analyzed using Cox regression. None of the patterns was significantly associated with the risk of NHL in the total population. However, the Vegetables pattern was inversely related to risk in Caucasian women with a hazard ratio of 0.56 (P(trend) = 0.04), and the Fat and Meat pattern was associated with a fivefold higher risk of follicular lymphoma in men (P(trend) = 0.03). The lack of significant results in men and women indicates a limited role of diet in NHL etiology, but dietary patterns might have ethnic- and subgroup specific effects on NHL. PMID- 19811331 TI - Immunophenotyping of mature B-cell non Hodgkin lymphoma involving bone marrow and peripheral blood: critical analysis and insights gained at a tertiary care cancer hospital. AB - We evaluated the diagnostic utility of flow cytometry immunophenotyping in bone marrow aspirates and peripheral blood, in the assessment of mature B-cell non Hodgkin lymphoma (MBNHL). We analyzed 356 cases of MBNHL received for immunophenotyping over a 4 year period. All cases were reviewed, correlated with biopsy specimen (lymph node and splenectomy). Discrepant cases were re-evaluated. Common subtypes included chronic lymphocytic leukemia (CLL) (243 cases, 68.5%), follicular lymphoma (30 cases, 8.5%), mantle cell lymphoma (20 cases, 5.5%), splenic marginal zone lymphoma (18 cases, 5%), hairy cell leukemia (18 cases, 5%). CD5+/CD23+ had a high positive predictive value (PPV) for diagnosing CLL whereas CD5+/CD23- had a high negative predictive value (NPV) for diagnosing mantle-cell lymphoma (MCL). Limited panel of 9 antibodies mainly CD19, CD5, CD23, CD10, FMC7, kappa, lambda, CD3 and CD20 help diagnose more than 92% of cases of MBNHL. Minimal diagnostic panels become important in countries with limited resources. PMID- 19811332 TI - Lack of prognostic value of MMP-9 expression and immunohistochemically defined germinal center phenotype in patients with diffuse large B-cell lymphoma treated with modern chemotherapy with or without CD20 antibody. AB - Diffuse large B-cell lymphomas (DLBCLs) are a heterogeneous group of lymphomas, with no accepted biological prognostic markers in routine clinical practice. Previously, matrix metalloproteinase (MMP-9), tissue inhibitors of matrix metalloproteinase (TIMP)- 1 and non-germinal center (GC) phenotype have been shown to associate with poor prognosis in DLBCL patients. The aim of this study was to find out whether tissue expression of gelatinases (MMP-2 and MMP-9) or their tissue inhibitors (TIMP-1 and TIMP-2) or immunohistochemically defined GC phenotype could act as prognostic markers in patients treated with modern treatments. Additionally, correlations between these proteins and GC phenotype were investigated. GC phenotype and tissue expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 were analyzed by immunohistochemistry in tissue samples from 114 DLBCL patients. In this study, in patients treated with modern lymphoma treatments (5 year cause-specific survival 69.8%) MMP-2, MMP-9, TIMP-1 or TIMP-2 expression or GC phenotype did not correlate with survival. International Prognostic Index (IPI) and stage were the only factors, which retained their prognostic significance in this patient material. Gelatinases or TIMPs did not correlate with GC phenotype, either. Prognostic markers are dependent on the lymphoma treatments used. In DLBCL patients treated with modern chemotherapy with or without rituximab, MMP-9, TIMP-1 and GC phenotype seem to have lost their prognostic value. PMID- 19811333 TI - Wilms' tumor gene 1 expression analysis by real-time quantitative polymerase chain reaction for monitoring of minimal residual disease in acute leukemia. AB - Wilms' tumor gene 1 (WT1) gene expression was analyzed in 32 patient with acute myeloid leukemia (AML) and 18 with acute lymphoblastic leukemia (ALL) to investigate whether it could serve as a MRD marker. Ninety-four percent of patients with acute leukemia showed high WT1 expression at presentation. WT1 expression as a MRD marker was evaluated in 36 patients. The rise of WT1 expression preceded the hematological relapse by approximately 4 months (mean time 129 days; range 6-298). The prognostic significance of WT1 expression was analyzed in 30 patients with AML. WT1 expression higher than 20 WT1 copies /10(4)ABL copies after induction and consolidation chemotherapy was associated with shorter OS. WT1 expression analysis could be a useful tool for MRD monitoring in acute leukemia. PMID- 19811334 TI - Meta-analysis study of glutathione-S-transferases (GSTM1, GSTP1, and GSTT1) gene polymorphisms and risk of acute myeloid leukemia. AB - To investigate the association of glutathione-S-transferase (GST) polymorphisms with the risk of acute myeloid leukemia (AML), a meta-analysis of case-control studies published between 1998 and 2009 was performed. Pooled odds ratios (ORs) were assessed using both fixed- and random-effects models. Heterogeneity across studies was calculated, and funnel plots were constructed to test for publication bias. Overall, the random-effects OR with GSTM1 null genotype, GSTP1 Val105 allele and GSTT1 null genotype were 1.30 (95% confidence intervals (CI) 1.04 1.62, p = 0.018), 1.03 (95% CI 0.80-1.33, p = 0.80) and 1.24 (95% CI 0.98-1.58, p = 0.06), respectively. Statistically, significant increased risk of AML was observed with GSTM1 while borderline significance was seen with GSTT1 null genotypes. However, fixed-effects model showed significant risk of AML in the presence of null genotypes of GSTM1 and GSTT1(p < 0.05). Significant heterogeneity was found between studies relating to GSTP1 (p = 0.162), however, no heterogeneity was seen in studies that evaluated GSTM1 (Q-value = 44; I(2) = 70.9; p-value < 0.01]; and GSTT1 (Q-value = 26.03; I(2) = 57.74; p-value < 0.01] polymorphisms. From the limited studies on the association of GSTP1 with risk of AML, the role of this gene cannot be ascertained fully. Significant association of these three genes with risk of AML must be evaluated further with respect to population, smoking, eating habits, ethnicity, and race. PMID- 19811335 TI - Increased serum bilirubin level without jaundice in patients with monoclonal gammopathy. PMID- 19811337 TI - Propofol sedation for upper gastrointestinal endoscopy in patients with liver cirrhosis as an alternative to midazolam to avoid acute deterioration of minimal encephalopathy: a randomized, controlled study. AB - OBJECTIVE: Midazolam sedation for upper gastrointestinal (GI) endoscopy exacerbates minimal hepatic encephalopathy (HE) in patients with liver cirrhosis, therefore an alternative drug regimen for these patients is warranted. The aim of this randomized, controlled study was to assess whether the use of the short acting propofol as a sedative for GI endoscopy could prevent the exacerbation of minimal HE in patients with liver cirrhosis. MATERIAL AND METHODS: The study comprised patients with liver cirrhosis without clinical HE who had undergone upper GI endoscopy for therapeutic purposes (intended variceal band ligation). Sixty patients were randomly assigned into two groups to receive propofol (n=40) or midazolam (n=20) for upper GI endoscopy. The study groups were matched for age, gender and Child-Pugh score. All patients completed number connecting tests (NCTs), as well as a porto-systemic encephalopathy (PSE) syndrome test before and at 2 h after completion of the endoscopic procedure. Time needed to fulfill the tests was documented. Baseline results of the psychomotor test batteries were compared with the post-interventional evaluations. Data were also compared with the results of a healthy control group (n=20) that did not undergo endoscopic sedation. Recovery time and quality (score system) were evaluated. RESULTS: The differences in the NCT times before and after sedation (median delta NCT, midazolam group, 11 s (95% CI, -1.2 to 16.1 s) versus the propofol group, -9.5 s (95% CI, -15.7 to -4.6 s), p=0.002) and in the PSE scores (median delta PSE, midazolam group, -1 (95% CI, -1.5 to 0.2) versus the propofol group, 1 (95% CI, 0.5 to 1.5), p=0.0009) differed significantly between the two groups. In addition, the recovery time and quality in patients receiving propofol were significantly improved compared with in the midazolam group (7.8+/-2.9 min versus 18.4+/-6.7 min, 6.1+/-1.1 versus 8.2+/-1.3, both p<0.001). CONCLUSIONS: The study demonstrates that propofol sedation for upper GI endoscopy does not cause acute deterioration of minimal hepatic encephalopathy and is associated with improved recovery in patients with liver cirrhosis. Propofol should be recommended for these patients as an alternative to midazolam. PMID- 19811340 TI - Biliary sphincterotomy does not relate to diarrhoea or major changes in bile acid synthesis or plasma lipids. AB - OBJECTIVES: Bile acid (BA) malabsorption may occur after cholecystectomy. Bile may flow more freely into the duodenum after endoscopic sphincterotomy (EST), in part resembling the situation after cholecystectomy. The (75)SeCHAT test used to diagnose BA malabsorption correlates inversely with synthesis and faecal excretion of BAs. The BA intermediate 7alpha-hydroxy-4-cholesten-3-one (C4) mirrors BA and lathosterol cholesterol synthesis. The aim was to study whether EST causes BA diarrhoea and alterations in BA synthesis or lipid profiles. MATERIAL AND METHODS: Twelve patients underwent the (75)SeHCAT test prior to and 3 months after undergoing EST and a further 22 only after EST. The Gastrointestinal Symptom Rating Scale (GSRS), 1 week daily stool frequency and consistency, C4, lathosterol, cholesterol and triglycerides were investigated. The (75)SeHCAT values of 29 healthy subjects served as controls. RESULTS: Stool frequency (median 1/day, IQR (interquartile range): 0.7) and consistency (median: 3, IQR: 0.65) were normal and none reported diarrhoea after EST (n=34). The GSRS scores were normal. There was no significant change in (75)SeHCAT (median 22%, IQR 29% versus 19.5%, IQR 25, n=12). There was a trend towards lower (75)SeHCAT after EST compared with the controls (median 26%, IQR 32, n=34 versus median 38%, IQR 19.5, n=29, p=0.075) and higher lathosterol (median 47.1 mg/mole, IQR 32.7 versus median 52.5 mg/mole, IQR 35.6, n=14, p=0.055). The C4 and lipids did not change significantly. CONCLUSIONS: EST did not induce diarrhoea and in line with this BA synthesis and serum lipids are unaltered. PMID- 19811342 TI - Predicting the clinical response to cytapheresis in steroid-refractory or dependent ulcerative colitis using contrast-enhanced ultrasonography. AB - OBJECTIVE: To evaluate the usefulness of transabdominal ultrasound (US), including contrast-enhanced ultrasonography (CEUS), in predicting the response to cytapheresis therapy in patients with steroid-refractory or -dependent ulcerative colitis (UC). MATERIAL AND METHODS: Between January 2005 and June 2008, 26 consecutive patients with steroid-refractory or -dependent UC were treated with granulocyte and monocyte adsorption apheresis (GCAP) or leukocytapheresis (LCAP) at our institute. The clinical activity of UC was evaluated by patients' C reactive protein (CRP) levels and clinical activity index (CAI) scores. All patients were evaluated by grey-scale US, power Doppler US (PDUS), and CEUS. In CEUS, the color signal patterns were classified as 1 of 2 patterns. In pattern 1, color signals were partially detected in the bowel wall (excluding muscularis propria, the outer thin layer of the bowel wall), whereas in pattern 2, color signals were detected in the entire bowel wall (excluding muscularis propria). Differences between remission or clinical response (group R) and no response (group N) were ascertained for clinical features, clinical activities, and US findings. RESULTS: Differences between the two groups were not considered significant for the clinical features, clinical activities, and grey-scale US and PDUS findings. Using CEUS, 4 patients in group R showed pattern 2 (21%), while in group N, all patients showed this pattern, indicating a significant difference between the two groups (p<0.01). CONCLUSION: CEUS findings may be helpful in predicting the clinical response to cytapheresis for steroid-refractory or dependent UC. PMID- 19811343 TI - Metformin in patients with non-alcoholic fatty liver disease: a randomized, controlled trial. AB - OBJECTIVE: The antidiabetic agent metformin is regularly discussed as a promising treatment for non-alcoholic fatty liver disease (NAFLD), which is characterized by insulin resistance. However, the evidence for its beneficial effects is limited, and conflicting reports have been published. The purpose of this study was to conduct a randomized, double-blind, placebo-controlled trial to test whether metformin improves liver histology in patients with non-alcoholic fatty liver disease. MATERIAL AND METHODS: Forty-eight patients with biopsy-proven NAFLD were randomized to treatment with metformin (n=24) or placebo (n=24) for 6 months. A second liver biopsy was obtained in all subjects who completed the trial (n=44). Data analyses are restricted to this group (per-protocol analyses). The primary outcome was changes in histologically assessed liver steatosis. Secondary outcomes were changes in NAFLD activity (NAS)-score, liver steatosis assessed by computed tomography (CT), liver transaminases, body-weight, metabolic variables and inflammatory markers. RESULTS: No significant differences between treatment with metformin or placebo were observed for changes in liver steatosis, assessed either histologically or by CT, NAS-score, liver transaminases or on markers of insulin resistance or inflammation. In contrast, beneficial effects of metformin were observed on changes in body-weight (p<0.001), serum levels of cholesterol (p=0.004), LDL-cholesterol (p<0.001), glucose (p=0.032) and on HbA1c (p=0.020). CONCLUSIONS: Treatment with metformin for 6 months was no better than placebo in terms of improvement in liver histology in patients with NAFLD. Nevertheless, the use of metformin could still be beneficial in this group as it is associated with a reduction in serum levels of lipids and glucose. (ClinicalTrials.gov number, NCT00303537). PMID- 19811344 TI - Abstracts of the 44th Annual Scientific Meeting of the Society for the Hypertension Related Disease Model Research, November 21-22, 2008, Izumo, Japan. PMID- 19811349 TI - Genetic variation in the renin-angiotensin system and arterial stiffness. The Rotterdam Study. AB - We studied the associations of three renin-angiotensin system polymorphisms, angiotensin-converting enzyme (ACE) I/D, angiotensinogen 235 M/T, and angiotensin II receptor type I 573 C/T, with arterial stiffness. The study was embedded in the Rotterdam Study, a population-based study older adults. The association of the polymorphisms with pulse wave velocity, the carotid distensibility, and pulse pressure was investigated in 3706 subjects. We found no association of the ACE I/D polymorphism with pulse wave velocity, but the D-allele was associated with a lower distensibility coefficient (p = 0.05) and higher pulse pressure (p = 0.01). For the angiotensinogen 235 M/T polymorphism, no significant associations with either pulse wave velocity (p = 0.71), the distensibility coefficient (p = 0.16) or pulse pressure (p = 0.34) were found. Also, we found no significant associations of pulse wave velocity (PWV) (p = 0.32), the distensibility coefficient (p = 0.08), and pulse pressure (p = 0.09) with the angiotensin II receptor type 1 573 C/T polymorphism. No epistatic effects were observed between the three renin-angiotensin system (RAS) genes with arterial stiffness. Our findings suggest that genetic variation in the renin-angiotensin system may play a role in determining carotid distensibility and pulse pressure. PMID- 19811350 TI - Influences of hypertension and diabetes on normal age-related changes in left ventricular function as assessed by tissue Doppler echocardiography. AB - Although the impact of hypertension (HT) and type 2 diabetes mellitus (DM) on left ventricular (LV) function has recently been studied using tissue Doppler echocardiography (TDE), there are few studies discriminating between the impact of the disease and that of normal aging on LV function. The purpose of the present study was to elucidate the LV function in patients with HT and DM in various age strata in order to assess the independent roles of HT and DM on normal age-related changes in cardiac function. The population of the study consisted of four groups: 20 control subjects (Control), 20 patients with hypertension alone (HTN), 20 patients with type 2 diabetes alone (DM), and 20 patients with both hypertension and diabetes (HTN+DM) in each of five age strata the 40s, 50s, 60s, 70s, and 80s. The strain and strain rate, which reflected both LV systolic and diastolic function, were assessed by TDE. The strain and strain rate decreased with advancing age in healthy control subjects and in all the patient groups. The strain and strain rate in the HTN group and the DM group showed lower values than those in the healthy control subjects in each age stratum. Furthermore, the strain and strain rate in the HTN+DM group showed the lowest values among all four groups in each age stratum. These results indicate that LV function as assessed by TDE demonstrates age-related deterioration with normal aging. Although HT or DM affects normal age-related changes in LV function, the co-existence of HT and DM has a more harmful effect on the normal age-related changes than HT alone or DM alone. PMID- 19811351 TI - Simultaneous determination of losartan and hydrochlorothiazide in human plasma by LC/MS/MS with electrospray ionization and its application to pharmacokinetics. AB - A method based on a simple liquid-liquid extraction (LLE) followed by high performance liquid chromatography with negative ion electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) detection was developed for the simultaneous determination of losartan (LOS) and hydrochlorothiazide (HCTZ) in human plasma, using valsartan (VAL) and chlorthalidone (CHTD) as an internal standard, respectively. The acquisition was performed in multiple reactions monitoring (MRM) and the limit of quantification was 4 ng/mL for both LOS and HCTZ. The method was linear in the studied range (4-800 ng/mL for LOS and 4-500 ng/mL for HCTZ). The intra-assay precisions ranged from 2.6-11.9% for LOS and 1.4 8.2% for HCTZ, while the inter-assay precisions ranged from 1.0-8.0% for LOS and 2.5-7.7% for HCTZ. The intra-assay accuracies ranged from 91.3 to 107.6% for LOS and 91.5 to 105.8% for HCTZ, while the inter-assay accuracies ranged from 99.9 to 106.4% for LOS and 97.4 to 101.4% for HCTZ. The analytical method was applied to a bioequivalence study, in which 28 healthy adult volunteers (14 men) received single oral doses (100 mg LOS + 25 mg HCTZ) of reference and test formulations, in an open, two-period, balanced randomized, crossover protocol. Based on the 90% confidence interval of the individual ratios for Cmax and AUC0-inf, it was concluded that the test formulation is bioequivalent to the reference Hyzaar formulation with respect to the rate and extent of absorption of both LOS and HCTZ. PMID- 19811352 TI - Lack of an association of GNB3 C825T polymorphism and blood pressure in patients with rheumatoid arthritis. AB - G-protein beta 3 subunit (GNB3) C825T (rs5443) single nucleotide polymorphism (SNP) has been implicated as a risk factor for essential hypertension in the general population. The effects of this SNP may be more prominent in subjects with endothelial dysfunction (ED). Rheumatoid arthritis (RA) is associated with ED and has a high prevalence of hypertension. Thus far, this SNP has not been studied in RA patients. We genotyped 383 RA patients and 432 controls. GNB3 C825T was identified using real-time polymerase chain reaction (PCR) and melting curve analysis. There were no differences in the frequencies of the GNB3 C825T genotype and alleles between RA and controls. Within RA patients, prevalence of hypertension did not differ across genotypes. The TT versus CC+CT contrast yielded an adjusted odds ratio (OR) of 0.92 (95% CI: 0.49 to 1.76, p = 0.813), the contrast of TT+CT versus CC an adjusted OR of 2.17 (95% CI: 0.885 to 5.30, p = 0.091), whereas that of the T allele versus C allele an adjusted OR of 1.11 (95% CI: 0.76 to 1.61, p = 0.604). Systolic and diastolic blood pressure levels were not significantly different across the three genotypic groups. No significant interaction was observed between GNB3 825C/T polymorphism and serum endothelin levels. Data from the present study suggest that the T825 variant of the G protein beta 3 subunit gene is unlikely to constitute major susceptibility loci for essential hypertension in Caucasian RA patients. Further larger studies are required to confirm our findings and assess the interaction of rs5443 with environmental factors. PMID- 19811353 TI - Hypotensive effect of Chamaemelum nobile aqueous extract in spontaneously hypertensive rats. AB - This study aims to evaluate the hypotensive effect of Chamaemelum nobile aqueous extract (CNAE) in spontaneously hypertensive rats (SHRs). Single oral administration of CNAE (140 mg/kg) produced a significant reduction (p < 0.05) in systolic blood pressure (SBP) after 24 h of the administration. Daily oral administration of CNAE (140 mg/kg) during 3 weeks produced a significant reduction in SBP in the day 8 (p < 0.01) of treatment. Furthermore, CNAE produced a significant increase in urinary output and electrolytes excretion (p < 0.01) from the day 8 to the end of treatment. We conclude that CNAE possesses a hypotensive and diuretic effect in SHR. PMID- 19811354 TI - Effect of high fat loading in Dahl salt-sensitive rats. AB - Salt sensitivity of blood pressure (BP) is speculated to be a characteristic in obesity-induced hypertension. To elucidate the influence of obesity on salt sensitive hypertension, we examined the effect of fat loading on BP, renal damage, and their progression induced by salt excess in Dahl salt-sensitive (S) rats. High fat (HF: 45% fat diet: 8 weeks) diet increased BP with greater weight gain and visceral fat accumulation than low fat (10% fat) diet. In HF-fed rats, plasma glucose, plasma insulin, and urinary catecholamine increased, and urinary protein tended to be elevated. Moreover, excessive salt (8% salt diet: 8 weeks) induced hypertension and proteinuria was accelerated in HF-fed rats. Therefore, fat loading increased BP in Dahl S rats possibly through insulin-resistance and sympathetic excitation. Moreover, fat loading accelerated salt-induced BP elevation and renal damage, suggesting excessive intake of both fat and salt, such as a civilized diet, exert the synergic harmful effects. PMID- 19811355 TI - Early diabetes in WKY and SHR produces decrease of the responses to angiotensin II and 5-HT and changes in the NO-GMPc pathway. AB - Early stages of diabetes have been related to arterial impairment in the vasoconstriction to norepinephrine. For that reason, the aim of this work was to investigate possible changes in the reactivity to angiotensin II and 5-HT in pithed rats and to evaluate the responses of aortic rings to acetylcholine and sodium nitroprussiate in streptozotocin-induced diabetes with 4 weeks evolution in both Wistar-Kyoto (WKY) and Spontaneously hypertensive rats (SHR). Our results suggest that hypertension produces a greater decrease in the vasoconstrictor response to angiotensin II and 5-HT in early stages of diabetes, while the NO GMPc pathway could be involved such effect. PMID- 19811356 TI - ACE I/D and G2350A polymorphisms in Pakistani hypertensive population of Punjab. AB - Several studies in different populations have shown mixed association of hypertension with ACE I/D and G2350A polymorphisms. To assess the link of these two polymorphisms with hypertension in population of Punjab--most populated province of Pakistan--we carried out this retrospective case control study in a mixed sample of 344 hypertensive and normotensive controls. Genotype of the ACE I/D was determined by nested PCR and G2350A. polymorphism was determined by amplification of a small fragment containing SNP and digesting it with the restriction enzyme. Statistical analysis revealed that I/D polymorphism is not associated with hypertension in the Punjabi population chi(2) (df = 2) = 5.611, P or = 65 years). According to the Japanese guidelines, goal LDL cholesterol (LDL-C) levels were defined based on the patient category. Average BP level and the number of anti-hypertensive drugs were 133 + or - 12/74 + or - 9 mmHg and 2.1 + or - 1.1, respectively, and the LDL-C level was 119 + or - 27 mg/dl. Goal BP was achieved in 40% of the patients of < 65 years and 67% of the elderly patients. Goal LDL-C was achieved in 65% of the patients. Even in the patients taking lipid-lowering agents (n = 178), 30% failed to achieve goal LDL-C levels. In the patients who achieved BP < 130/85 mmHg, 67% also achieved goal LDL C, whereas 61% of the patients whose BP > or = 140/90 mmHg achieved goal LDL-C. Both goal BP and LDL-C were achieved in 39% of the male and 36% of the female patients. In contrast, neither goal BP nor goal LDL-C was achieved in 16% of the male and 17% of the female patients. Results suggest that intensive intervention should be required to achieve satisfactory BP and lipid control in hypertensive patients. PMID- 19811359 TI - White coat effect and its clinical implications in the elderly. AB - The aim of this study was to investigate the frequency and correlated factors of white coat effect (WCE) in the elderly. Geriatric patients who were known as normotensive and office BP exceeding 140/90 mmHg underwent 24-hour ambulatory blood pressure monitoring (ABPM). Correlation of WCE with clinical parameters, geriatric assessment scales, co-existing diseases, and laboratory results were analyzed. Within 61 patients 72.1% were diagnosed as white coat hypertension (WCH). Independent correlates of systolic WCE were activities of daily living, instrumental activities of daily living scores, creatinine; independent correlate of diastolic WCE was Geriatric Depression Scale score. White coat hypertension constitutes a major part of office-detected hypertension in geriatric patients. Ambulatory blood pressure monitoring should be performed on geriatric patients with office-measured hypertension in order to avoid overtreatment. PMID- 19811360 TI - Is prehypertension really different from normotension and hypertension? A case control pilot proteomic study in Chinese. AB - This study was designed to investigate the differences in the plasma proteome of healthy normotensive, prehypertensive, and hypertensive subjects. A case-control pilot study was conducted among well-matched subjects. Plasma protein was analyzed by two-dimensional electrophoresis combined with MALDI-TOF mass spectrometry. The results showed that there were 22 differentially expressed spots among the three groups, which corresponded to 18 proteins involved in inflammation and immunity, lipid metabolism, transport, coagulation and fibrinolysis, cell proliferation and apoptosis, and anti-oxidation. With an increase in blood pressure, these proteins were differentially expressed which indicated that prehypertension probably was an early stage of hypertension. PMID- 19811361 TI - Spectral analysis of heart rate variability for early prediction of pregnancy induced hypertension. AB - The early prediction of pregnancy-induced hypertension (PIH), a common morbid disorder of pregnancy is unsatisfactory. Therefore, in the present study we have investigated the role of spectral analysis of heart rate variability (HRV) in the early prediction of PIH. Spectral analysis of HRV was performed in three groups of subjects (Group I: normal pregnant women; Group II: pregnant women with risk factors, but did not develop PIH; Group III: pregnant women with risk factors and developed PIH). It was observed that the LF-HF ratio, the most sensitive indicator of sympathovagal balance, was significantly high (p < 0.01) since early pregnancy in group III compared to other groups, which was significantly correlated with heart rate and blood pressure. It was suggested that the predictive knowledge of sympathovagal imbalance should be utilized in designing the prevention and management of PIH. PMID- 19811362 TI - Anti-hypertensive effect of gamma-aminobutyric acid (GABA)-rich Chlorella on high normal blood pressure and borderline hypertension in placebo-controlled double blind study. AB - The anti-hypertensive effect of GABA-rich Chlorella was studied after oral administration for 12 weeks in the subjects with high-normal blood pressure and borderline hypertension in the placebo-controlled, double-blind manner in order to investigate if GABA-rich Chlorella, a dietary supplement, is useful in control of blood pressure. Eighty subjects with Systolic blood pressure (SBP) 130-159 mmHg or diastolic blood pressure (DBP) 85-99 mmHg (40 subjects/group) took the blinded substance of GABA-rich Chlorella (20 mg as gamma-aminobutyric acid) or placebo twice daily for 12 weeks, and had follow-up observation for an additional 4 weeks. Systolic blood pressure in the subjects given GABA-rich Chlorella significantly decreased compared with placebo (p < 0.01). Diastolic blood pressure had the tendency to decrease after intake of GABA-rich Chlorella. Neither adverse events nor abnormal laboratory findings were reported throughout the study period. Reduction of SBP in the subjects with borderline hypertension was higher than those in the subjects with high-normal blood pressure. These results suggest that GABA-rich Chlorella significantly decreased high-normal blood pressure and borderline hypertension, and is a beneficial dietary supplement for prevention of the development of hypertension. PMID- 19811363 TI - The functional changes of the perivascular adipose tissue in spontaneously hypertensive rats and the effects of atorvastatin therapy. AB - The aim of this study was to examine the function of perivascular adiposa tissue (PVAT) on vascular relaxation response in spontaneously hypertensive rats (SHR) and the modulatory effects of the atorvastatin therapy on the PVAT functions. We investigated the mechanisms of the perivascular adipocyte-derived relaxation factor (PVRF) by using isolated rat's aortic rings and isometric contraction measurements. We found that contraction of the thoracic aorta induced by phenylephrine was significantly attenuated in the presence of PVAT from normotensive Wistar-Kyoto rats (WKY group) or the spontaneously hypertensive rats treated with atorvastatin (SHR-A group, atorvastatin 50mg/kg/day), whereas this effect was not observed in the thoracic aortic rings from the control SHR (SHR group). Transferring the solution incubated with PVAT-intact thoracic aorta to PVAT-free thoracic aorta, it induced a remarkable relaxation response in the WKY but not in the control SHR. Tetraethylammoniumchloride (TEA) could block the above relaxation. It was also shown that the PVRF function was likely, depending on the extracellular [Ca(2+)]; the anti-contractile effect of PVAT could be reduced by the inhibitor of the adenosine triphosphate (ATP)-dependent potassium channels, glibenclamide, and could be reduced by the inhibitor of cyclooxygenase by indomethacin. We thus infer that the PVAT function was distorted in hypertension rats, and the lipid-lowering treatment with atorvastatin could restore the PVAT function. The function of the PVRF may involve the Ca(2+) activated potassium channels, the ATP-dependent potassium channels in vascular smooth muscle cell (SMC), and the release of PVRF from PVAT may involve prostaglandins (PGs) and the calcium metabolism. These results provide an insight into the pathological mechanisms of hypertension development, and indicate that the PVAT may be a potential new target for the hypertensive therapy. PMID- 19811364 TI - Regular exercise produced cardioprotective effects on rat's heart with hypertension induced by L-NAME administration. AB - This study investigated the effects of exercise training on apoptosis, angiotensin II expression, and eNOS expression in rats' heart with hypertension induced by N (omega)-nitro-L-arginine methylester (L-NAME) in vivo. Forty rats were divided into control (C), L-NAME treated (L), L-NAME treated with exercise (L + Ex), and exercise (Ex) groups. Systolic (S) and diastolic (D) blood pressures (BP) were elevated from the first week in L. In L + Ex, SBP and DBP were decreased from the fourth and second week, respectively. In L + Ex, decreased apoptosis and angiotensin II expression and increased eNOS expressions were detected. Consequently, exercise reduces BP by alternation of angiotensin II expression and eNOS expression, and inhibits apoptotic changes in the heart. PMID- 19811365 TI - The hypertensiogenetic steroid 19-nor-progesterone does not influence cortisol inactivation by 11beta-hydroxysteroid dehydrogenase type 2. AB - 19-nor-progesterone (19-nor-P) has the characteristics of a potent mineralocorticoid in adrenalectomized or salt-loaded rats and is capable of causing hypertension. In human placenta, progesterone is converted to 19-hydroxy progesterone, a precursor of 19-nor-P. In some states of pregnancy hypertension, 19-nor-P may inhibit renal 11beta-hydroxysteroid dehydrogenase type 2 (11beta HSD2), thus allowing cortisol to bind to the mineralocorticoid receptor (MR). Therefore, we investigated the ability of 19-nor-P to inhibit human 11beta-HSD2. Fetal kidney cells (HEK 293) were transfected with human 11beta-HSD2 and incubated with increasing concentrations of 19-nor-P, labelled and unlabelled cortisol. Steroids were extracted, separated by TLC, and radioactivity was measured using a TLC scanner. 19-nor-P treatment did not significantly reduce 11beta-HSD2 activity (430 to 300 pmol/mg protein/h) in the range of tested concentrations. In conclusion, 19-nor-P did not inhibit human 11beta-HSD2 and seems not to be involved in human hypertension. Nevertheless, 19-nor-P may be converted by extra-adrenal tissues into 19-nor-deoxycorticosterone (DOC) or 19 nor-corticosterone, which are potent mineralocorticoids and may be involved in the pathogenesis of hypertension during pregnancy. PMID- 19811366 TI - Coronary flow velocity reserve and aortic distensibility indices in hypertensive patients with hypercholesterolaemia and normal epicardial coronary arteries. AB - The aim of the present study was to assess the relationship between coronary flow velocity reserve (CFR), aortic distensibility indices, and levels of plasma total cholesterol (TC) in arterial hypertension. We studied 39 consecutive hypertensive patients free of coronary artery disease. Eighteen of them had normal TC and 21 had 1(8) high TC. Coronary flow velocity reserve and aortic elastic moduli were measured during stress transoesophageal echocardiography. Reduced CFR (2.41 + or 0.75 vs. 1.97 + or - 0.60, p < 0.05), increased aortic elastic modulus (in 10(3) mmHg) (0.56 + or - 0.41 vs. 0.77 + or - 0.29, p < 0.05) and Young's modulus (in 10(3) mmHg) (5.42 + or - 4.03 vs. 8.57 + or - 4.79, p < 0.05) were found in hypercholesterolaemia. PMID- 19811367 TI - Augmentation therapy in alpha-1 antitrypsin deficiency. PMID- 19811368 TI - COPD and coronary heart disease: challenges in understanding the natural history of common complex chronic diseases. PMID- 19811369 TI - Average age at death from COPD in the United States: 1980-85, 1990-95, 2000-05. AB - COPD represents an important public health challenge, in the US and globally, that is both preventable and treatable. We describe the average age at death from COPD, a leading cause of death in the US, using data from the National Vital Statistics System for the periods 1980-85, 1990-95, and 2000-05. Average age at death from COPD increased 3-4 years between 1980-85 and 2000-05 for men and women as well as for Whites and Blacks. PMID- 19811370 TI - Cardiac troponin-I predicts long-term mortality in chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease patients are at increased risk for mortality, particularly from cardiovascular conditions. Acute exacerbation increases heart burden and may lead to release of troponin I. This study investigates the long-term prognostic value of elevated troponin I detected during acute exacerbation of chronic obstructive pulmonary disease. The records of 182 patients with acute exacerbation in whom troponin I levels were sampled during their hospitalization were reviewed retrospectively. Receiver operator curve was used to determine the cut-off level for troponin I that discriminated survivors and non-survivors, and predictors for all-cause mortality were tested in a multivariate analysis. The results showed that, during a mean observation time of 50.1 +/- 45.6 months, 66 (36.3%) patients died, providing 1, and 3-year survival rates of 84%, and 54%, respectively. Troponin I levels were significantly higher in non-survivors compared with survivors, mean troponin I +/ SD in microg x L(-1): 1.35 +/- 3.17 vs. 0.53 +/- 2.08, respectively, p = 0.0033. ROC curve analysis identified troponin I > 0.03 microg x L(-1) as the optimal cut off level for prediction of mortality. Kaplan-Meier survival analysis revealed that the probability of survival was significantly lower in patients with troponin I > 0.03 microg x L(-1) (log-rank test p = 0.0058). On multivariate analysis, only ischemic heart disease (HR = 2.335, p = 0.0017) and troponin I level (HR = 1.31541, p = 0.2513) were independent predictors of mortality. In conclusion, it was found that a mildly elevated troponin I level measured in patients with chronic obstructive pulmonary disease during acute exacerbation is a strong independent predictor of mortality following discharge. PMID- 19811371 TI - Distal airway function assessed by oscillometry at varying respiratory rate: comparison with dynamic compliance. AB - Distal airways disease causes significant morbidity yet remains insufficiently identified. We hypothesize that: ( [1] ) when spirometry is normal impulse oscillometry may provide information about mechanical properties of the distal airways in a manner comparable to dynamic compliance and ( [2] ) variation of breathing frequency will influence oscillometric measurements similar to effects of breathing frequency on dynamic compliance. Fifty-three symptomatic subjects with normal large airway function (spirometry) were studied; distal airway dysfunction was identified by presence of frequency dependence of compliance (FDC). Oscillometric parameters evaluated were resistance at 20 Hz (R20) and 5-20 Hz (R(5-20)), reactance at 5 Hz (X5), and reactance area (AX). R20 correlated with airway resistance by esophageal manometry (r = 0.74, p < 0.001); X5 correlated with dynamic compliance at a respiratory rate of 60 bpm (r = 0.61, p < 0.001); R(5-20) and AX correlated with FDC (r = 0.48, p < 0.001; r = 0.53, p < 0.01). IOS indices were further evaluated at increased respiratory rate (RR40). Oscillometric parameters changed minimally at RR40 in normal subjects DeltaR20 = 0.20 +/- 0.08 cmH2O/L/s, DeltaR(5-20) = 0.10 +/- 0.03 cmH2O/L/s, DeltaAX = 0.33 +/- 0.19 cmH2O/L). However, in symptomatic subjects, while R20 increased minimally at RR40 (DeltaR20 = 0.37 +/- 0.10 cmH2O/L/s), R(5-20) and AX increased markedly (DeltaR(5-20) = 0.54 +/- 0.06 cmH2O/L/s, DeltaAX = 4.28 +/- 0.67 cmH2O/L) and reversed post bronchodilator. IOS evaluates physiology of the distal airways in a manner comparable to dynamic compliance. Elevated respiratory rate influences oscillometric parameters and must be considered when interpreting oscillometric data. IOS provides a non-invasive tool for assessment of distal airway function when spirometry is normal, which can be applied to various clinical settings including early diagnosis of COPD (GOLD stage 0), asthma in clinical remission and occupational/ environmental irritant exposure. PMID- 19811372 TI - Non-invasive ventilation (NIV) in the clinical management of acute COPD in 233 UK hospitals: results from the RCP/BTS 2003 National COPD Audit. AB - Non-invasive ventilation (NIV) is a clinically proven, cost-effective intervention for acidotic exacerbations of COPD that is recommended by UK national guidelines. This study examines the extent to which these recommendations are being followed in the UK. Between August and October 2003 a national audit of COPD exacerbations was conducted by the Royal College of Physicians and the British Thoracic Society. 233 (94%) UK hospitals submitted data for 7,529 prospectively recruited acute COPD admissions, documenting process of care and outcomes from a retrospective case note audit. They also completed a resources and organisation of care proforma. Nineteen hospitals (8%) reported they did not offer NIV. There was no access to NIV in 92 (39%) intensive care units in 88 (36%), high-dependency units or on general wards of 85 (34%) hospitals. In 74 (30%) NIV was available on all 3 sites. A low pH (<7.35) was noted at some time during admission for 26% (1714/6544) of patients and NIV was administered to 31%. Patients receiving NIV were more often admitted under a respiratory physician, or seen at some stage by a respiratory specialist and had more severe disease (higher PaCO2 (median 9.8 v 7.8 kPa), lower oxygen tension (median 8.8 v 9.8 kPa), higher incidence of peripheral oedema (48% v 39%), of pneumonia (27% v 16%), higher in-hospital mortality (26% v 14%) and at 90 days (37% v 24%) and longer hospital stays (median 9 v 7 days) than those not receiving NIV. Hospitals with least usage of NIV had similar mortality rates to those using NIV more often. A comprehensive NIV service is not available in many hospitals admitting patients with acute respiratory failure secondary to COPD. Access to acute NIV is inadequate and does not conform with NICE and BTS guidelines. These observational audit data do not demonstrate benefits of NIV on survival when compared to conventional management, contrary to results from randomised trials. Reasons for this are unclear but unmeasured confounding factors and poor patient selection for NIV are likely explanations. PMID- 19811373 TI - Augmentation therapy for alpha1 antitrypsin deficiency: a meta-analysis. AB - BACKGROUND: Augmentation with exogenous alpha1-antitrypsin (alpha1-AT) is the only specific therapy for alpha1-AT deficiency. Uncertainty persists concerning its effectiveness. PURPOSE: To test the hypothesis that augmentation therapy in patients with alpha1-AT deficiency slows the decline in FEV1. STUDY SELECTION: Randomized and nonrandomized clinical studies with either parallel-group design or single cohort pre-post design were eligible if they compared augmentation therapy with a control regimen and if long-term (> 1 y) longitudinal FEV1 follow up data were collected. DATA SYNTHESIS: FEV1 data from five trials with 1509 patients were combined by random effects meta-analysis. The decline in FEV1 was slower by 23% (absolute difference, 13.4 ml/year; CI, 1.5 to 25.3 ml/year) among all patients receiving augmentation therapy. This overall protective effect reflected predominantly the results in the subset of patients with baseline FEV1 30-65% of predicted. In that subset, augmentation was associated with a 26% reduction in rate of FEV1 decline (absolute difference, 17.9 ml/year; CI, 9.6 to 26.1 ml/year). Similar trends amongst patients with baseline FEV1 percent of predicted < 30% or > 65% were not statistically significant. CONCLUSIONS: This meta-analysis supports the conclusion that augmentation can slow lung function decline in patients with AAT deficiency Patients with moderate obstruction are most likely to benefit. PMID- 19811374 TI - Roflumilast increases Clara cell secretory protein in cigarette smoke-exposed mice. AB - Decreased Clara cell secretory protein (CCSP) levels have been found in smokers and chronic obstructive pulmonary disease (COPD) patients, which may be related to the development of COPD. A phosphodiesterase-4 (PDE4) inhibitor, roflumilast, appears to have therapeutic value for COPD. However, its effect on CCSP in cigarette smoke (CS)-exposed lungs has not been investigated. AKR/J mice were treated as follows: air control, CS, roflumilast plus CS, and roflumilast. Mice underwent four weeks of air or CS exposure. Roflumilast was administrated at 5mg/kg via gavage once daily for the duration of the study. CCSP levels in bronchoalveolar lavage (BAL) fluid and ERK1/2 activation in lungs were examined. CS exposure tended to decrease CCSP levels in BAL fluid compared to air controls. Treatment with roflumilast significantly reversed CS-induced downward trend of CCSP in BAL fluid. Roflumilast significantly inhibited CS-induced upward trend of ERK1/2 activation in lungs, and the levels of activated ERK1/2 in lungs negatively correlated with CCSP levels of BAL fluid in CS, and CS plus roflumilast groups. Our results demonstrate that one of the therapeutic mechanisms of roflumilast is to reverse CS-induced downward trend in CCSP levels of BAL fluid, which may be mediated by down-regulating ERK1/2 activity. PMID- 19811375 TI - Activities of life: the COPD patient. AB - Chronic obstructive pulmonary disease (COPD) is a chronic, progressive and debilitating disease. With worsening disease, a COPD patient will progressively become less physically active, will have reduced social contacts, and this will create a downward vicious circle. This review discusses the significant sequelae of physical inactivity in the COPD patient, approaches to assessment and management, and the impact of COPD exacerbations on the activities of daily life. Patients with COPD are markedly inactive in daily life. Referral to pulmonary rehabilitation and self-management education to promote increase physical activities should be an important part of the management of COPD. Acute exacerbations in COPD patients are severely distressing events that impact greatly on health status, loss of symptom control and functional state with limitation on activities of daily living. If we are to progress in the treatment of acute exacerbation of COPD, it is evident that the effects of interventions on activities of daily living should be considered as an essential component of our goal of treatment. PMID- 19811378 TI - Long-term oxygen therapy: whatever happened to transtracheal oxygen? PMID- 19811376 TI - Mesenchymal cell fate and phenotypes in the pathogenesis of emphysema. AB - Emphysema is characterized by the destruction of alveolar parenchymal tissue and the concordant loss of lung epithelial cells, endothelial cells, and interstitial mesenchymal cells. Key features in the pathobiology of emphysema include inflammation, alveolar epithelial cell injury/apoptosis, and excessive activation of extracellular matrix (ECM) proteases. Mesenchymal cells are versatile connective tissue cells that are critical effectors of wound-repair. The excessive loss of connective tissue and the destruction of alveolar septae in emphysema suggest that the mesenchymal cell reparative response to epithelial injury is impaired. Yet, the mechanisms regulating mesenchymal cell (dys)function in emphysema remain poorly understood. We propose that mesenchymal cell fate, modulated by transforming growth factor beta-1 (TGF-beta1) and the balance of ECM proteases and antiproteases, is a critical determinant of the emphysema phenotype. We examine emphysema in the context of wound-repair responses, with a focus on the regulation of mesenchymal cell fate and phenotype. We discuss the emerging evidence supporting that genetic factors, inflammation and environmental factors, including cigarette smoke itself, collectively impair mesenchymal cell survival and function, thus contributing to the pathogenesis of emphysema. PMID- 19811377 TI - Defining disease modification in chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a debilitating condition characterized by airflow limitation that is not fully reversible. It is a major cause of morbidity and mortality and represents substantial economic and social burden throughout the world. A range of interventions has been developed that decrease symptoms and address complications associated with COPD. However, to date few interventions have been unequivocally demonstrated to modify disease progression. Assessment of the potential for interventions to modify disease progression is complicated by the lack of a clear definition of disease modification and disagreement over appropriate markers by which modification should be evaluated. To clarify these issues, a working group of physicians and scientists from the USA, Canada and Europe was convened. The proposed working definition of disease modification resulting from the group discussions was "an improvement in, or stabilization of, structural or functional parameters as a result of reduction in the rate of progression of these parameters which occurs whilst an intervention is applied and may persist even if the intervention is withdrawn". According to this definition, pharmacologic interventions may be considered disease-modifying if they provide consistent and sustained improvements in structural and functional parameters. Smoking cessation and lung volume reduction surgery would both qualify as disease-modifying interventions. PMID- 19811380 TI - Smoking and COPD-mechanisms and prevention. PMID- 19811381 TI - Reduced phosphorylation of the TGF-Beta signal transducer Smad2 in emphysematous human lung. AB - Chronic obstructive pulmonary disease (COPD) is characterized by inflammatory immune response, emphysematous destruction of alveolar structures and obstruction in the small conducting airways. Transforming growth factor (TGF)-beta is involved in the maintenance of normal lung tissue homeostasis as a regulator of extracellular proteolysis, tissue repair and inflammatory functions. This study was undertaken to characterize TGF-beta signaling in pathologically distinct areas of COPD lungs. Using Smad2 phosporylation (P-Smad2) as an indicator of TGF beta signaling activity we analyzed COPD patient tissues and controls by immunohistochemistry. Emphysematous lung showed significantly reduced P-Smad2 immunoreactivity both in the alveolar and bronchiolar epithelium, which is evidence of reduced TGF-beta signaling activity. On the contrary, in the thickened peribronchial areas there was an increase in the amount of P-Smad2 positive cells. Isolated COPD lung fibroblasts also displayed increased TGF-beta signaling and target gene expression suggesting that the fibroblasts are characteristic to the small airway disease phenotype. Our results indicate that TGF-beta signaling activity is differentially regulated in distinct areas of COPD lung and likely contributes to both emphysematous development and small airway obstruction. PMID- 19811382 TI - Long-term smoking behavior patterns predicting self-reported chronic bronchitis. AB - We examined the effects of long-term smoking patterns on self-reported symptoms of chronic bronchitis within the Finnish adult twin cohort including 21, 609 individuals responding to questionnaires in 1975 and 1981, of which 11,015 respondents participated also in 1990. We also explored the association between smoking and bronchitis among discordant twin pairs. Among those without chronic bronchitis at baseline we examined incidence of chronic bronchitis in 1981 both by 1975 smoking status, but also based on subgroups formed according to change in smoking behaviors between 1975 and 1981. We conducted similar analyses in the longitudinal data including three consecutive measurements of smoking status. Logistic regressions demonstrated that among current smokers, the risk of chronic bronchitis increased about 1.5-fold by each amount category of daily cigarettes. When analyzing change of smoking status between 1975 and 1981, daily moderate and heavy smokers, smoking increasers and decreasers, as well as re-current smokers demonstrated elevated risks. In the analysis among discordant twin pairs the smoking co-twins had a 14-fold likelihood for chronic bronchitis compared to their never-smoking co-twins. Panel analyses showed that, not only moderate and heavy, but also former and light smokers, had significant risks for chronic bronchitis. Those with late smoking initiation, leisure time physical activity or over 10 years of smoking cessation were less likely to have chronic bronchitis. We conclude that in long term evaluation no safe level of smoking exists. Abstinence from tobacco seems to be the public health message justified by these results in prevention of chronic bronchitis. PMID- 19811383 TI - Hypertension, systemic inflammation and body weight in relation to lung function impairment-an epidemiological study. AB - Recent reports on the simultaneous occurrence of systemic inflammation and airflow obstruction are usually based on a highly selective patient population, but their importance warrants further evaluation in the general population. The objectives were to study the interrelationship between airflow obstruction, smoking, hypertension, obesity and CRP as a marker of systemic inflammation in a randomly selected sample of the general Icelandic population (n = 939). This study comprised 758 randomly selected men and women 40 years and older living in Reykjavik, Iceland, and who were participating in the Burden of Obstructive Lung Disease (BOLD) study (81% response rate). In addition to the BOLD protocol, which included post-bronchodilator spirometry, they answered questions about general health and medication. Serum samples were taken for measurement of C-reactive protein (CRP). In the sample-245 individuals (33%) reported having hypertension. Subjects with hypertension were older, had a higher BMI and higher CRP levels. Subjects with hypertension had lower values of FEV(1) than predicted (89.9 +/- 18.5 vs. 94.5 +/- 14.4%) (p < 0.001) and FVC (92.2 +/- 15.1 vs. 95.3 +/- 12.3%) (p = 0.002). These differences remained significant after adjusting for age, BMI, CRP and smoking. Hypertension and CRP levels above the median were both independently and additively associated with lower FEV(1) and FVC. In addition a lower FVC% was also associated with a higher BMI (> 30 mg/m2). Use of betablocking antihypertensives was not related to lung function. Hypertension, BMI and systemic inflammation affect lung function independently of each other. All three variables have a negative effect on FVC, while hypertension and high CRP were independently associated with impaired FEV(1). PMID- 19811384 TI - Real-world clinical data identifies gender-related profiles in chronic obstructive pulmonary disease. AB - This study aims to compare diagnostic and clinical outcomes of Chronic Obstructive Pulmonary Disease (COPD) from the gender perspective using retrospective health care data and patient reported outcomes in a real-world setting. An electronic database was constructed from complete medical records of 844 COPD patients who were recruited in Helsinki and Turku University Central Hospitals during the years 2005-07. The patients were identified from the hospital discharge registries by ICD10 code J44.8 in the age group of 18-75 years of age. The medical history was obtained from all healthcare providers who had treated these patients during the previous 5-10 years; the study intends to continue their follow-up annually for the next 10 years. Thirty-six percent (N = 266) of the participants were women. The COPD diagnosis had been made at the same age for both genders. Women, however, reported significantly less pack-years than men. Compared to the men, the women displayed less advanced airway obstruction, but more severe gas transfer impairment. Parenchymal damage when evaluated by diffusion capacity correlated significantly stronger with FEV(1) (% of predicted) in women than men. The BMI index of the women was lower than that of the men. Cardiovascular diseases, diabetes and alcoholism were significantly more common in men, but women suffered more psychiatric conditions, especially depression. This cohort showed several significant gender dependent differences in the clinical presentation that need to taken under consideration in the assessment of COPD progression and the disease management. PMID- 19811385 TI - A 20-year follow-up of a population study-based COPD cohort-report from the obstructive lung disease in Northern Sweden studies. AB - Mortality and other long-term outcomes of COPD from epidemiological studies of cohorts based on the general population are still rare. In contrast, data from follow-ups of patients from hospitals and general practices are more common and demonstrate often a 5-year mortality of about 50% and even higher. The aim was to study 20-year outcomes, mainly mortality, in a COPD cohort derived from a population study. The Obstructive Lung Disease in Northern Sweden (OLIN) Study's first postal survey was performed in 1985, and 5698 subjects (86%) responded. A stratified sample of symptomatic subjects and controls was invited to clinical examinations including lung function tests in 1986, 1506 (91%) of the invited participated and 266 subjects fulfilled the GOLD criteria of COPD. All alive and possible to trace had participated at least at two follow-up examinations. Of the 266 subjects with COPD 46% were still alive after 20 years. The proportion of survived among subjects with severe and very severe COPD at entry was 19%. Death was significantly related to age, male sex, disease severity and concomitant ischemic heart disease or cardiac failure at entry. Socioeconomic status (manual workers) was significant in the univariate analysis, but failed to reach statistical significance in the multivariate model. The annual decline in FEV(1) among survivors was low to normal. Long-term follow-ups of subjects with COPD derived from population studies provide data reflecting the course of COPD in society better than follow-ups of hospital recruited patients, who represent the top of the iceberg. Surprisingly many with severe COPD were still alive after 20 years. PMID- 19811386 TI - Therapeutic strategies to optimize the efficacy of nicotine replacement therapies. AB - Smoking is estimated to be responsible for at least 2/3 of chronic obstructive pulmonary disease (COPD) deaths. Mortality rates due to all causes and to COPD decline progressively after smoking cessation compared with continuing smoking. Nicotine replacement therapies (NRT) increase the likelihood of smoking abstinence by only 60%. Optimization of NRT is of importance in COPD patients because they may be more nicotine dependent and have more difficulties to quit than smokers without COPD. The objective was to critically review pharmacotherapeutic strategies to optimize the efficacy of NRT. Findings revealed that fixed high dose NRT does not convincingly result in higher abstinence rate compared with standard dose and increases the likelihood of adverse effects in smokers with low need for nicotine. Combination of NRT of different routes of administration versus single NRT provides a statistically significant benefit over a single NRT. A 2-week treatment by nicotine patch before quit day approximately doubles post-quit day abstinence. NRT augmentation with burpropion or nortriptyline, antidepressants with demonstrated efficacy for smoking cessation, does not seem to ameliorate further abstinence rates. Three months' and 6 months' NRT exposure was compared by only one but sufficiently powered study and found similar abstinence rates. Optimization strategies to increase the efficacy of NRT include combining NRT of different routes of administration and use of nicotine patch before target quit day. Uncertainties exist about the optimal length of NRT administration. Co-administration of NRT with bupropion or nortriptyline does not seem to lead to higher abstinence rate than NRT alone. PMID- 19811387 TI - The multiple facets of cigarette addiction and what they mean for encouraging and helping smokers to stop. AB - Addiction involves powerful motivation to engage in an activity repeatedly to an extent that is harmful often accompanied by impaired capacity for self-control. To effectively combat addiction to cigarettes requires an understanding that there are several mechanisms underlying it. The PRIME Theory of motivation aims to provide a model that can encapsulate these mechanisms. It recognises that evolution has led to multiple levels of motivation from basic impulses and inhibitions, through 'motives' (feelings of want and need), to 'evaluations' (beliefs about what is good or bad), and plans (intentions regarding future actions). Self-control involves self-consciously generating motives from evaluations or plans; it requires and depletes mental energy. Nicotine from cigarettes generates the motivation to smoke and undermines self-control by interacting with all of the level of motivation. It: creates stimulus-impulse associations resulting in cue-driven urges; impairs inhibitory control; gives enjoyment resulting in 'wanting' to smoke; it leads to 'nicotine hunger', withdrawal symptoms and beliefs about benefits of smoking (e.g. stress relief) all of which can result in a 'need' to smoke. Evidence is emerging that wanting to smoke (because of enjoyment) is a major deterrent to making quit attempts but does not influence success, while cue-driven impulses to smoke, nicotine hunger and adverse mood and beliefs about the benefits of smoking are important in relapse. Combating cigarette addiction requires attention to all of these factors. PMID- 19811388 TI - The role of dendritic cells in the pathogenesis of COPD: liaison officers in the front line. AB - Dendritic cells are professional antigen presenting cells linking innate and adaptive immune responses. Different dendritic cell subsets were identified in human lung, each with their own functional characteristics. As innate and adaptive immune responses are activated in Chronic Obstructive Pulmonary Disease (COPD), dendritic cells could play a role in the pathogenesis of this disease. Indeed, cigarette smoke appears to modulate dendritic cell function in vitro and alters dendritic cell numbers and function in cigarette smoke exposed mice. The number of pulmonary dendritic cells differs between COPD patients, smokers and non-smokers. Moreover, the number of Langerhans-type dendritic cells increases with the severity of the disease. In this review we will discuss the scientific evidence regarding the role of dendritic cells in COPD and we will put forward the concept of modulation of dendritic cell differentiation and function as a crucial step in the pathogenesis of COPD. PMID- 19811389 TI - Current perspectives on role of chromatin modifications and deacetylases in lung inflammation in COPD. AB - Chromatin modifications and epigenetic regulation are critical for sustained and abnormal inflammatory response seen in lungs of patients with chronic obstructive pulmonary disease (COPD) because the activities of enzymes that regulate these epigenetic modifications are altered in response to cigarette smoke. Cigarette smoke induces chromatin modifications and epigenetic changes by causing post translational modifications of histone acetyltransferases, and histone/non histone deacetylases (HDACs), such as HDAC2 and sirtuin 1 (SIRT1), which leads to chromatin remodeling. In this review, we discussed the current knowledge on cigarette smoke/oxidants-induced post-translational modifications of deacetylases (HDAC2 and SIRT1), disruption of HDAC2/SIRT1-RelA/p65 corepressor complex associated with acetylation of RelA/p65, and chromatin modifications (histone H3 phospho-acetylation) leading to sustained pro-inflammatory gene transcription. Knowledge on molecular mechanisms of epigenetic changes in abnormal lung inflammation will help in understanding the pathophysiology of COPD which may lead to the development of novel epigenetic therapies in the near future. PMID- 19811390 TI - From proteomics to prescription-the search for COPD biomarkers. AB - Biomarkers that describe the severity and progression of COPD and the responses of patients to treatment are a desirable addition to clinical measures of disease. In this review, we describe the current state of knowledge of biomarkers used for the diagnosis, staging and therapeutic response of COPD patients. The nature of these biomarkers is considered in relation to their intended use, and the desirable qualities of such entities are examined. Examples of biased and unbiased discovery platforms for COPD biomarker discovery are given, and the major findings of these studies are discussed. Cutting edge technology used for biomarker discovery, quantitation in biofluids and imaging biomarkers in whole body systems is reviewed. PMID- 19811391 TI - Genetic epidemiology of smoking behavior and nicotine dependence. AB - The genetic epidemiology of smoking behavior and nicotine dependence is reviewed. Twin, family and adoption studies show consistent evidence for genetic effects on many aspects of smoking behavior and nicotine dependence, while molecular genetic analyses have only recently identified genes consistently associated with nicotine dependence and amount smoked. Further studies with more sophisticated phenotypes, larger sample sizes and better measures of the environment are needed. PMID- 19811392 TI - Extracellular superoxide dismutase and risk of COPD. AB - Chronic obstructive pulmonary disease (COPD) is a major public health problem worldwide. COPD is strongly related to cigarette smoke exposure, but not all smokers develop the disease. It is thought that COPD progresses slowly over time stimulated by environmental exposures, including free radicals from cigarette smoke, which ultimately establish chronic inflammation and result in a progressive destruction of lung tissues. COPD is known to occur in family clusters, which has prompted interest in determining genetic risk factors for the disease. Several genetic studies have identified an association between extracellular superoxide dismutase (ECSOD) polymorphisms and risk for developing COPD. ECSOD is an antioxidant protein that scavenges superoxide free radicals from cigarette smoke and protects the lungs from free radical damage and chronic inflammation. PMID- 19811394 TI - Biophysical cancer transformation pathway. AB - Coherent vibration states in biological systems excited in nonlinear electrically polar structures by metabolic energy supply were postulated by H. Frohlich. Frohlich's requirements for coherent vibrations and generation of electromagnetic field are satisfied by microtubules whose subunits are electric dipoles. Static electric field around mitochondria and "wasted energy" efflux from them provide nonlinear conditions and coherent excitation. Numerical models are used for analysis of coherent vibration states. A hypothesis is presented that dysfunction of mitochondria (i.e., extinction of the zones of the static electric field and of the efflux of "wasted energy") and disintegration of the cytoskeleton on the pathway of cancer transformation result in disturbances of coherence of the cellular electrically polar oscillations and of the generated electromagnetic field with consequences in cellular organization and interactions between cells. Local invasion, detachment, and metastasis of cancer cells are subsequent events of disturbed electromagnetic interactions. PMID- 19811395 TI - Electric polarization and the viability of living systems: ion cyclotron resonance-like interactions. AB - Wellness can be described in physical terms as a state that is a function of the organism's electric polarization vector P(r, t). One can alter P by invasive application of electric fields or by non invasive external pulsed magnetic fields (PMF) or ion cyclotron resonance (ICR)-like combinations of static and sinusoidal magnetic fields. Changes in human (total) body bioimpedance are significantly altered during exposure to ICR magnetic field combinations. The conductivities of polar amino acids in solution exhibit sharp discontinuities at ICR magnetic fields tuned to the specific charge to mass ratio of the amino acid. It has been reported that protein peptide bonds are broken by such amino acid ICR fields. Remarkably, some of these effects are only found at ultra-low AC magnetic intensities, on the order of .05 muT. This is approximately 10(3) below accepted levels determined by engineering estimates. Such strikingly low magnetic intensities imply the existence of physically equivalent endogenous weak electric field oscillations. These observations not only make claims related to electromagnetic pollution more credible but also provide a basis for future electromagnetic applications in medicine. They also reinforce the notion that physical factors acting to influence the electric polarization in living organisms play a key role in biology. PMID- 19811396 TI - Do field-free electromagnetic potentials play a role in biology? AB - All bio-systems are imperfect dielectrics. Their general properties however cannot be described by conventional simple electrodynamics; the system is more complex. A central question in our present paper is centered on a controversial debate of the possible effect of the zero fields (only potentials exist). We show that the identical use of the "field-free," "curl-free," and "force-free" terminologies is incorrect, there have definitely different meanings. It is shown that the effective electro-dynamical parameters that describe and modify living systems are the potentials and not the fields. We discuss how the potentials have a role in biological processes even in field-free cases. PMID- 19811397 TI - Oncothermia treatment of cancer: from the laboratory to clinic. AB - Oncothermia is a long-time applied method (since 1989) in oncology. Its clinical results excellently show its advantages, however the details of its mechanism are under investigation even today. The method is based on a self-selective process of energy concentration and targets the membrane of the malignant cell, using the temperature gradient and the beta-dispersion of the membrane proteins. To prove the theory we show the experimental evidences in vitro experiments where we showed the definite difference between the conventional heating and the oncothermia at the same temperature. In the next step, we studied some xenograft nude-mice models, verifying the temperature-dependent and non temperature dependent factors. In addition, the synergic effect with some chemotherapies were studied, having more efficacy of the oncothermia with drugs than the conventional heating. These experiments show the definite advantages of the oncothermia compared to its classical counterpart, acting on the same temperature. We have also proved the beneficial effect of oncothermia treatment in the veterinary practice Oncothermia is applied in numerous clinics and hospitals, and we would like to show some characteristic case-reports and also the clinical benefit on the survival time elongation of liver-, pancreas-, brain-, and lung-tumor lesions. PMID- 19811398 TI - Effect of curl-free potentials on water. AB - Living objects are complex systems with various harmonized chemical, thermodynamical, and quantum-mechanical processes in aqueous electrolyte environment. We had studied the effect of curl-free magnetic vector-potential on the matrix of the living matter, on the water. The discussed theoretical considerations are in harmony with the presented simple experiments. It is shown that the vector-potential is actually an effective electro-dynamical parameter which could modify the processes in living systems. PMID- 19811399 TI - Inapplicability of small-polaron model for the explanation of infrared absorption spectrum in acetanilide. AB - The applicability of small-polaron model for the interpretation of infrared absorption spectrum in acetanilide has been critically reexamined. It is shown that the energy difference between the normal and anomalous peak, calculated by means of small-polaron theory, displays pronounced temperature dependence which is in drastic contradiction with experiment. It is demonstrated that self-trapped states, which are recently suggested to explain theoretically the experimental absorption spectrum in protein, cannot cause the appearance of the peaks in absorption spectrum for acetanilide. PMID- 19811400 TI - Metal, EMF, and brain energy metabolism. AB - Some implications of cooperative potential of metal ions and electromagnetic fields' radiation (EMF) in carcinogenic processes are discussed. It is known that these factors, chemical and physical individually have connections with processes of oxidative stress. Special attention was paid to possible manifestation within the brain. Therefore, the entry of a few potentially neurotoxic metals into the brain is discussed. PMID- 19811401 TI - Occupation-dependent access to metabolic energy in Frohlich systems. AB - Frohlich systems model ensembles of electromagnetic-radiation-emitting molecular oscillators residing inside biological cells. The core piece of this theory are rate equations for the quantum-mechanical occupation numbers of the collective oscillation modes that take into account the flow of metabolic energy through the ensemble. In continuation of prior study of feedback patterns implicit in this model, oriented multigraphs representation is used to investigate the case when the pumping source-in analogy with the effect of spectral hole burning in injection lasers-exhibits depletion phenomena dependent on occupation numbers and the eigenfrequencies of the modes. As a result, in comparison with the case when all modes have equal access to pumping source, the onset of energy condensation in the fundamental mode is shifted to higher pumping levels and new feedback loops appear in the representative diagram. The modified theory provides a more realistic description of electromagnetic emission in biological cells. PMID- 19811402 TI - Electromagnetic fields and the induction of DNA strand breaks. AB - The International Agency for Research on Cancer (IARC) has classified the extremely low-frequency (ELF) electromagnetic fields (EMF) as "possible carcinogenic" based on the reported effects. The purpose of this work is to review and compare the recent findings related to the induction of DNA strand breaks (DNA-SB) by magnetic field (MF) exposure. We found 29 studies (genotoxic and epigenetic) about the induction of DNA-SB by MF. 50% showed effect of MF and 50% showed no DNA-SB. Nevertheless, considering only genotoxic or only epigenetic studies, 37.5% and 69.2% found induction of DNA-SB by MF, respectively. In relation to these data it seems that MF could act as a co-inductor of DNA damage rather than as a genotoxic agent per se. Nevertheless, the published results, in some cases conflicting with negative findings, do not facilitate to obtain a common consensus about MF effects and biophysical interaction mechanisms. PMID- 19811403 TI - Blood-brain barrier disruption by continuous-wave radio frequency radiation. AB - The increasing use of cellular phones and the increasing number of associated base stations are becoming a widespread source of non ionizing electromagnetic radiation. Some biological effects are likely to occur even at low-level EM fields. This study was designed to investigate the effects of 900 and 1,800 MHz Continuous Wave Radio Frequency Radiation (CW RFR) on the permeability of Blood Brain Barrier (BBB) of rats. Results have shown that 20 min RFR exposure of 900 and 1,800 MHz induces an effect and increases the permeability of BBB of male rats. There was no change in female rats. The scientific evidence on RFR safety or harm remains inconclusive. More studies are needed to demonstrate the effects of RFR on the permeability of BBB and the mechanisms of that breakdown. PMID- 19811404 TI - Measurement of electrical oscillations and mechanical vibrations of yeast cells membrane around 1 kHz. AB - Frohlich postulated coherent polar oscillations as a fundamental biophysical property of biological systems. Recently, Pelling et al. (2004, 2005) detected mechanical vibrations of yeast cell membrane with atomic force microscope (AFM) and analyzed by Fourier analysis in the frequency range 0.5-2 kHz with amplitudes of the order of 1 nm. This article describes the measurement of electric activity of yeast cells in the acoustic frequency range and of mechanical vibrations of cell membrane. Spectrum analyzer and electrically and electromagnetically screened box with point sensor and amplifiers fed by batteries were used for measurement of synchronized and non synchronized tubulin mutants of yeast cells. We show that the electric activity of synchronized cells in the M phase is greater that of non synchronized cells. That corresponds to the findings of Pohl et al. (1981). Obtained results of measurement of cell electric activity are in good agreement with AFM findings. PMID- 19811405 TI - Autologous Hsp70 induces antigen specific Th1 immune responses in a murine T-cell lymphoma. AB - Heat Shock protein-70 derived from tumor cells is highly immunogenic and induces specific anti-tumor immune response by directly activating cytotoxic CD8(+) T cells. Additionally, Hsp70 is known to be a strong activator of antigen presenting cells and therefore, up regulates the production of pro-inflammatory cytokines and chemokines. In this study, we have shown the effect of tumor derived Hsp70 on the induction of delayed type hypersensitivity reaction in a T cell lymphoma bearing mice. The autologous Hsp70 augments contact hypersensitivity and delayed type hypersensitivity responses in mice challenged with allergen in vehicle and antigens respectively. The adoptive transfer of splenocytes derived from Hsp70 immunized mice is able to enhance delayed type hypersensitivity response in antigen challenged normal and DL-bearing host. Furthermore, adoptive transfer of macrophages incubated with autologous Hsp70 also enhances DTH reactivity in mice. The pro-inflammatory cytokines and C-C chemokines are found to be elevated in the DTH footpad extract of antigen challenged normal and DL-bearing mice. Increased production of IFN-gamma and MIP 1alpha+/- suggest that autologous Hsp70 augments the recruitment of antigen specific Th1 cells, which further secretes pro-inflammatory cytokines and C-C chemokines mediating the hypersensitivity reaction upon challenge with antigens. PMID- 19811406 TI - In vivo proliferation of rat lamina propria T lymphocytes: general hyporesponsiveness but increased importance of the CD2 and CD28 pathways. AB - Lamina propria T lymphocytes (LPL-T) have a low proliferative potential in vitro. We asked whether LPL-T are also hyporesponsive in vivo and whether this is specific for the alphabeta T cell receptor (TCR). Mitogenic mAb directed at the alphabeta TCR, CD2, CD28, or control mAbs plus IL-2 were injected into rats. Proliferation and/or apoptosis were detected by double staining using 5-bromo-2' deoxyuridine/TUNEL and the alphabeta TCR. LPL-T were hyporesponsive to various stimuli compared to other T cells. The strongest proliferation was found upon CD2/CD28 stimulation (LPL-T: 281 +/- 6%; spleen: 642 +/- 31%). LPL-T proliferation was only detectable at 24 h while proliferation in other compartments also occurred later. Hyporesponsiveness was not caused by enhanced T cell apoptosis upon alphabeta TCR stimulation. In conclusion, stimulation of LPL T results in much shorter and weaker in vivo proliferation than in other lymphoid organs. Overall, CD2/CD28 costimulation is the strongest T cell stimulus in vivo. PMID- 19811407 TI - The essential oil isolated from Artemisia capillaris prevents LPS-induced production of NO and PGE(2) by inhibiting MAPK-mediated pathways in RAW 264.7 macrophages. AB - Artemisia capillaris (A. capillaris) is used in traditional Korean herbal medicine for its believedanti-inflammatory activities. Previous studies have suggested that the essential oil of A. capillaris contains the active components responsible for its pharmacological effect, even though the mechanism for its action is unclear. This study examined the inhibitory effects of the essential oil of A. capillaris on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)). The essential oil significantly inhibited the production of NO in the LPS-stimulated RAW 264.7 macrophages, which was mediated by the down-regulation of inducible NO synthase (iNOS) expression but not by its direct cytotoxic activity. The essential oil also blocked the secretion of PGE(2) and the expression of cyclooxygenase-2 (COX-2) in the LPS stimulated cells. Western blot analysis showed that the essential oil inhibited the phosphorylation of IkappaB-alpha, nuclear translocation of p65, and subsequent activation of NF-kappaB. In addition, the essential oil suppressed the LPS-stimulated activation of mitogen-activated protein kinases (MAPKs) as well as the AP-1 DNA-binding activity. Moreover, MAPK inhibitors significantly reduced the LPS-induced production of NO and PGE(2). Collectively, we suggest that the oil inhibits the expression and production of inflammatory mediators by blocking the MAPK-mediated pathways and inhibiting the activation of NF-kappaB and AP-1. PMID- 19811408 TI - Involvement of T(H)1/T(H)2 cytokines in the pathogenesis of autoimmune skin disease-Pemphigus vulgaris. AB - Pemphigus vulgaris (PV) is classical example of antigen-driven severe autoimmune bullous skin disorder. Auto reactive T cells are critical for the induction and regulation of antibody production. With regard to cytokine production profiles, it has been reported that qualitative as well as quantitative alterations in cytokine production can result in activation of inefficacious effector mechanisms and therefore, complex and severe impairment in immune functions. The purpose of this study was to observe the alterations in the levels of T(H)1 [Interleukin-2 (IL-2), Interferon-gamma (IFN-gamma)] and T(H)2 (IL-4 and IL-10) cytokines in the sera from patients affected with PV and compared with Pemphigus foliaceus and healthy subjects. This work is aimed to comprehend the involvement of T(H)1 and T(H)2 cells as inflammatory infiltrate in the modulation of acantholysis and production of pemphigus lesions. Seventy PV, 13 PF and 50 healthy, age-matched individuals without any generalized skin diseases were included in this study. The diagnosis of PV and PF patients was confirmed by histopathology (hematoxylin and eosin) and / direct immunofluorescence. The levels of T(H)1 cytokines (IL-2 and IFN-gamma) and T(H)2 cytokine markers (IL-4 and IL-10) were estimated by high sensitivity ELISA kits. All patients with PV and PF showed significantly (p < 0.000) elevated levels of T(H)2 cytokines (IL-10 and IL-4) as compared with healthy controls. However, the mean concentration of T(H)1 cytokines (IL-2 and IFN-gamma) was significantly decreased in patients as compared to healthy individuals. Both T(H)1 and T(H)2 cytokines did not show any significant difference between PV and PF cases. Current concepts support the idea that PV, induced by autoantibodies against Dsg3, is the consequence of an imbalance between Dsg3-reactive T(H)2 and T(H)1 cells that may be critical for the maintenance of tolerance against Dsg3. Cytokine profile for confirmed PV cases showed direct evidences for involvement of T cell responses. Increase in IL-4 and IL-10 shows induction of T(H)2 cells in the pathogenesis of autoimmune disorders Pemphigus vulgaris. The decreased levels of IL-2 and IFN-gamma might demonstrate the inhibitory effects by IL-4 and IL-10, which suppress the expansion of T(H)1 population. PMID- 19811409 TI - Development of an indirect enzyme-linked immunosorbent assay for the organophosphorus pesticide paraoxon-methyl. AB - In the present study, the synthesis of hapten for the organophosphorus (OP) pesticide paraoxon-methyl was developed, with a spacer arm (aminocarboxylic acid) attached at the aromatic ring. It was conjugated to bovine serum albumin (BSA) for use as an immunogen and to ovalbumin (OVA) for coating antigen for ELISA testing. Rabbits were immunized with the immunogen and two polyclonal antisera were produced and screened against the coating antigen using competitive indirect enzyme-linked immunosorbent assay (ELISA). For application to textile samples, the influence of several factors such as organic solvent, ionic strength, and pH on the ELISA results were studied. Under optimized conditions, the quantitative working range was 0.012-1.158 microg/mL with a limit of detection (LOD) of 0.005 microg/mL and the IC(50) was 0.115 microg/mL.There was negligible cross reactivity (CR) with other OP pesticides. The recoveries obtained by standard paraoxon-methyl addition to the different textile samples such as cotton, wool and muslin delaine were all from 86.0% to 108.0%. Therefore, the optimized ELISA may become a new convenient and economical analytical tool for monitoring paraoxon-methyl residues in textile samples. PMID- 19811410 TI - Proteomic analyses of the effects of drugs of abuse on monocyte-derived mature dendritic cells. AB - Drug abuse has become a global health concern. Understanding how drug abuse modulates the immune system and how the immune system responds to pathogens associated with drug abuse, such hepatitis C virus (HCV) and human immunodeficiency virus (HIV-1), can be assessed by an integrated approach comparing proteomic analyses and quantitation of gene expression. Two-dimensional (2D) difference gel electrophoresis was used to determine the molecular mechanisms underlying the proteomic changes that alter normal biological processes when monocyte-derived mature dendritic cells were treated with cocaine or methamphetamine. Both drugs differentially regulated the expression of several functional classes of proteins including those that modulate apoptosis, protein folding, protein kinase activity, and metabolism and proteins that function as intracellular signal transduction molecules. Proteomic data were validated using a combination of quantitative, real-time PCR and Western blot analyses. These studies will help to identify the molecular mechanisms, including the expression of several functionally important classes of proteins that have emerged as potential mediators of pathogenesis. These proteins may predispose immunocompetent cells, including dendritic cells, to infection with viruses such as HCV and HIV-1, which are associated with drug abuse. PMID- 19811411 TI - Effects of aging on antioxidant response and phagocytosis in senescent erythrocytes. AB - Red blood cell (RBC) aging is a complex process affected by immunological and biochemical parameters. In this work we studied the antioxidant response in RBC of different ages. We also investigated their interaction with peripheral blood monocytes. Anticoagulated blood samples from 19 O RhD+ volunteers' donors were processed. Young (Y) RBC and Senescent (Se) RBC were obtained by self-formed gradients of Percoll. The fractionation of the erythrocytes suspensions was demonstrated by statistically significant density-related changes in hematological determinations. Activities of glucose-6-phosphate dehydrogenase (G6PD), of soluble NADH-cytochrome b5 reductase (b5Rs) and membrane-bound b5R (b5Rm) were determined spectrophotometrically. The interaction between monocytes and different RBC suspensions was evaluated by the erythrophagocytosis assay. The G6PD and b5Rm activities in SeRBC were significantly lower than that observed in YRBC. No differences were found in the b5Rs of both groups. We observed an increased rate of erythrophagocytosis the SeRBC compared to YRBC. The decline in the activities of G6PD and b5Rm would indicate a decrease in the antioxidant response associated to RBC aging. These findings would signify that the oxidative changes of membrane occurring during the life span of the RBC might be relevant in the process of removal of SeRBC from the circulation. PMID- 19811412 TI - Evaluation of the contribution of major T cell subsets to IFN-gamma production in TB infection by ELISPOT. AB - Interferon gamma remains a key effector molecule that is still widely used as the most informative biomarker for screening human immune responses against tuberculosis, particularly in ELISPOT assays. We investigated the participation of CD4(+) and CD8(+) T lymphocytes in the PBMC responses to Mycobacterium tuberculosis (Mtb) specific antigens in 33 TB cases and 49 contacts. Responses to ESAT-6 were higher than CFP-10. There was no significant difference in responses to both Mtb antigens between cases and contacts. PBMCs response to ESAT-6 but not CFP-10 in cases was significantly reduced by depletion of CD4(+) cells whereas CD8(+) cell depletion had no impact. In conclusion, ESAT-6 is a more recognized antigen in this population, and CD4(+) lymphocytes are the main participants in IFN-gamma response by ELISPOT. Thus, a decline of CD4(+) T lymphocytes below a critical level might affect the sensitivity of IFN-gamma release assays for detecting Mtb infection. PMID- 19811413 TI - Decreased expression of T lymphocyte co-stimulatory molecule CD26 on invariant natural killer T cells in systemic lupus erythematosus. AB - CD26, a T cell co-stimulatory molecule and dipeptidyl peptidase IV for the degradation of interferon-gamma-induced chemokine, participates in multiple immunopathological roles in leukocyte homing and inflammation. Decreased circulating concentration of soluble (s)CD26 in patients with systemic lupus erythematosus (SLE), rheumatoid arthritis and murine model of arthritis and encephalomyelitis have been reported. In the present study, the plasma concentration of sCD26 and chemokines, and cell surface expression of CD26 on monocytes, CD4+T lymphocytes, CD8+T lymphocytes, CD19+B lymphocytes and invariant natural killer T (iNKT) lymphocytes were analyzed using ELISA and flow cytometry, respectively, in 23 SLE patients and 14 sex- and age-matched control subjects. Although there was no significant difference between plasma concentrations of soluble CD26 in SLE patients with controls (p > 0.05), there was significant elevated Th1 chemokines CXCL10 and CXCL9 but not Th2 chemokine CCL2, and down regulation in iNKT lymphocytes number and cell surface expression of CD26 on CD4+T and iNKT lymphocytes of SLE patients compared with controls (all p < 0.05). Decreased circulating number of iNKT cells and CD26 on iNKT cells can be important for the immunopathogenesis by exacerbating Th1-related inflammation in SLE. PMID- 19811414 TI - Dietary vitamin D3 restriction influences tumor growth, but not the ability to generate an antigen-specific immune response in OTII transgenic mice. AB - Initially, we wanted to know whether dietary vitamin D(3) restriction would influence growth and metastasis of the 4T1 murine mammary carcinoma. We confirmed serum 25(OH)D levels were modulated by dietary vitamin D(3) restriction, mice were healthy, and when challenged with the 4T1 tumor alterations in tumor growth, but not metastasis were evident. Tumors grew more rapidly in mice on the vitamin D(3) restricted diet. To delineate whether dietary vitamin D(3) restriction influenced the ability to generate an antigen-specific immune response we used OTII transgenic mice which express a T cell receptor specific for ovalbumin. We found that dietary vitamin D(3) restriction did not influence the health of OTII mice, the number of circulating CD3/CD4(+), CD3/CD8(+), CD4/CD25(+) T cells, nor the ability to generate CD11c(+) bone-marrow derived dendritic cells. T cells from OTII mice maintained on the vitamin D(3) restricted diet also exhibited no significant alterations in proliferative capacity or ability to secrete IFN-gamma or IL-4 in an antigen-specific manner. Yet, EL-4 tumors grew more rapidly in OTII mice on the vitamin D(3) restricted diet. These data show that dietary vitamin D(3) restriction impacts tumor growth, but not the ability to generate an antigen specific immune response. PMID- 19811415 TI - Alopecia areata universalis after phenobarbital-induced anti-convulsant hypersensitivity syndrome. AB - Alopecia is an adverse effect in those patients taking aromatic anti-convulsant drugs but is rarely reported after discontinuing such medications in the convalescent status of anti-convulsant hypersensitivity syndrome (AHS). A 3-year old boy developed alopecia areata (AA) universalis in the convalescent status of phenobarbital-induced AHS, compatible to the evidences of increased lymphocyte proliferation and increased dead cells percentages while his peripheral blood mononuclear cells were incubated with phenobarbital. Skin histology revealed peri follicular, peri-bublar and supra-bublar lymphocyte infiltration. By searching for the key words AHS, alopecia areata (AA, punctuate absence of terminal scalp hair), AA totalis (complete absence of terminal scalp hair), and AA universalis (total loss of terminal scalp and body hair) using PubMed, only 2 cases, to date, developed alopecia in the convalescent status of phenobarbital-induced AHS. Among these 3 cases, all had favorable prognosis despite having jaundiced hepatitis. Their hair grew back after 2-3 months steroid therapy. Alopecia does rarely develop in the convalescent status of phenobarbital-induced AHS after stopping phenobarbital and its mechanism is related to lymphocyte infiltration into the peri-bulbar, supra-bulbar and peri-follicular regions. PMID- 19811416 TI - Serum levels of soluble fas in patients with multinodular goiter. AB - OBJECTIVE: Fas is a cell-surface receptor responsible for induction of apoptosis in human thyrocytes upon interaction with Fas Ligand. Fas protein expression on thyroid cells and Fas-mediated apoptosis is decreased in multinodular goiter (MNG) resulting in thyroid cell proliferation. The soluble form of Fas (sFas) produced by alternative mRNA splicing may inhibit Fas-Fas Ligand binding and apoptosis. The aim of this study was to examine whether sFas is differentially expressed in multinodular goiter (MNG), which is associated with decreased Fas mediated apoptosis. METHOD: We determined serum sFas levels using enzyme-linked immunosorbent assay (ELISA) in 42 patients with MNG and 23 normal controls. RESULTS: Serum sFas levels were increased in patients with MNG (7.47 +/- 2.55 ng/ml) compared to normal controls (2.26 +/- 0.9 ng/ml). Levels of sFas were not significantly correlated with age, sex or clinical parameters, such as serum levels of FT4 or TSH. DISCUSSION: Increased sFas in MNG may indicate increased expression of alternatively spliced Fas mRNA variant and decreased expression of cell-surface Fas protein, and may enhance thyroid cell proliferation by protecting thyroid cells from Fas-mediated apoptosis. PMID- 19811417 TI - Soluble PD-1 is associated with aberrant regulation of T cells activation in aplastic anemia. AB - Engagement of the membrane program death-1 (PD-1) receptor by its ligands suppresses T cell proliferation and cytokine production. Aberrant over-expression of costimulatory molecules, including PD-1, has been associated with persistent activation of self-reactive T cells in autoimmune diseases. However, the mechanism underlying the dysfunction of PD-1 in the regulation of T-cell activation in such diseases remains unclear. Here, we report the overexpression of CD4(+) and CD8(+) T cell PD-1 and elevated serum levels of soluble PD-1 in aplastic anemia (AA) patients. Detailed characterization of soluble PD-1 revealed that it corresponded to an alternative splice variant PD-1Deltaex3, which lacks the transmembrane domain but has a soluble extracellular domain of the PD-1 molecule. In a further study, PD-1 fusion protein displayed the ability of increasing the proliferation of T cells in vitro, which suggested that soluble PD 1 might serve as an autoimmune antibody to block the function of membrane-bound PD-1 on T cells and lead to aberrant T cell proliferation. Our study revealed a novel pathogenic pathway in which the function of overexpressed PD-1 to restrict over-self-reaction is counteracted by the excessive production of soluble PD-1. PMID- 19811418 TI - Epitope mapping of monoclonal antibody 1B9 against plasmodium falciparum-derived macrophage migration inhibitory factor. AB - A homologue of mammalian macrophage migration inhibitory factor (MIF) has been identified in Plasmodium falciparum (PfMIF). This parasite-derived cytokine and its antiserum were detected in the circulation of patients with malaria. Using a monoclonal antibody, designated mAb1B9, against PfMIF, we performed biopanning using two phage display peptide libraries to screen for the main sequence of the epitope recognized by the antibody. We then expressed a series of truncated peptides in order to identify the precise sequence of the epitope. The epitope recognized by mAb1B9 is 22 amino acids long and has the following sequence: (36)LGYIMSNYDYQKNLRFGGSNEA(57). Western analysis showed that the residues (36)LG(37) and (52)G differentiated PfMIF from the rodent malaria parasite derived MIFs, and the residues (43)Y and (48)NL(49) differentiated PfMIF from P. vivax- and P. knowlesi-derived MIFs. The precise identification of this epitope, the first identified for PfMIF, will increase the specificity of the sandwich ELISA assays used to evaluate patients with malaria. These results indicate that mAb1B9 is useful for investigating the function of PfMIF in immune responses to malaria. Both the epitope and the monoclonal antibody against it will be valuable tools in epidemiological studies concerning this P. falciparum-derived cytokine. PMID- 19811419 TI - Characteristics of expanded CD4+CD28null T cells in patients with chronic hepatitis B. AB - Expanded CD4(+)CD28(null) T cells have not been described in the circulation of patients with chronic hepatitis B (CHB). The aim of the present was to detect and characterize the surface phenotype and functional capacity of these cells in CHB patients. Expanded CD4(+)CD28(null) T cells were detected in the circulation of CHB patients with high viral load and elevated aminotransferase levels. Most CD4(+)CD28(null) T cells showed a CD27-CD45RA(-)CD45RO(+) surface phenotype. The markers CD56, CD57 and killer immunoglobulin-like receptor (KIR) were detected on CD4(+)CD28(null) T cells, but the majority were positive for CD57. Functionally, CD4(+)CD28(null) T cells were found to be potent cytotoxic T lymphocytes with perforin and granzyme B secretion profiles. These findings indicate that the expanded CD4(+)CD28(null) T cells are cytotoxic memory T cells and display a distinct functional phenotype in comparison with CD4(+)CD28(+) T cells. The presence of these cells appears to be associated with inflammatory conditions, suggesting that these elevated CD4(+)CD28(null) T cells might be involved in the pathogenesis of CHB. PMID- 19811421 TI - Allotransplantation of sulphate glucomannan-alginate barium (SGA) microencapsulated rat islets for the treatment of diabetes mellitus. AB - To offer a more effective microencapsulation technique of islets for the treatment of diabetes, we have developed a new type of microcapsule comprising sulphate glucomannan-alginate barium (SGA). We compared it with traditional microencapsulated APA (alginate-poly-L-lysine-alginate) and ABa (Ba(2+)-alginate) microencapsulated islets. These three types of microencapsulated islets were prepared and cultured in vitro and we studied their morphology and activity. To determine their effects on insulin secretion and cytokine production (MCP-1, IL 1, IFN-gamma, TNF-alpha) the islets were transplanted into diabetic rats. There was no difference in the morphologies of the three types of microencapsulated islets or their insulin secretory capacity in vitro. However, the SGA microencapsulated islets had higher activity and produced more insulin than the APA and ABa microencapsulated islets after transplantation. Normoglycemia was maintained for longer in the SGA-transplanted group than in the other two groups. The concentrations of cytokines in the peritoneal fluid were significantly decreased in the SGA group, as was the infiltration of inflammatory cells around the microcapsules. In conclusion, the novel SGA microencapsulated islets can maintain normoglycemia in diabetic rats without immunosuppression for longer than APA and ABa microencapsulated islets. PMID- 19811422 TI - Asymmetric IgG antibodies induced by different immunotherapies in a murine model of allergy. AB - Specific immunotherapy (SIT) is the only potentially curative treatment for those allergic processes mediated by IgE. We compared the effects of different SITs in mice sensitised with ovalbumin (OVA) Al (OH)(3) : 1) OVA entrapped in particles of poly (D,L-lactic-co-glycolic acid) (PLGA-OVA), 2) Soluble OVA (OVA-sol) and 3) Polymerised OVA (OVA-pol). Serum levels of specific IgE, IgG1, IgG2a and asymmetric IgG, the cutaneous anaphylaxis test (PCA), and the IL-10, IFNgamma and IL-4 levels in culture supernatants of splenocytes challenged with OVA were assessed. Mice treated with PLGA-OVA had higher levels of asymmetric antibodies than non-desensitised mice; a low IgG1 and high IgG2a level was observed together with inhibitory effect in the PCA reaction that reversed in the absence of asymmetric IgG. IL-10 and IFNgamma levels were higher in supernatants from mice treated with PLGA-OVA and OVA-sol than those obtained from non-desensitised controls. Our results suggest that among the different SITs evaluated, PLGA-OVA is the one that best showed an increase in the asymmetric IgG molecules and an effective deviation of the immune response. Furthermore, the increase in the proportion of asymmetric antibodies would be of importance when designing new vaccination strategies for allergy. PMID- 19811423 TI - Rapid immunosorting of transmembrane proteins of Lymphocytes from a cDNA expression library of COS-1 cells. AB - The proteins on lymphocyte surface play important roles in a wide range of immunological processes, but the profile and characterization of surface proteins remain to be further investigated, among which the method for fast screening of surface proteins needs to be established. In this study, a conventional cDNA clone library of hepatic lymphocytes from C57BL/6 mouse was constructed, and then the cDNA was inserted into a recombinant expression vector pSecTag-attR with a signal peptide and tag protein for fluorescence screening. The recombinant cDNA expression library was transfected into COS-1 cells, and the transfected cells with the expressed membrane proteins were labeled by fluorescence antibodies and isolated by fluorescence activated cell sorting. After two cycles of sorting, the purity of fluorescence positive cells with membrane proteins was up to 98%, and the representative membrane molecules on lymphocytes such as CD3, CD4, CD8, NK1.1 and NKG2D were detected in the library. These results demonstrated that the cDNA expression library containing transmembrane proteins provided an efficient and fast tool for the study of transmembrane proteins on hepatic lymphocytes. PMID- 19811424 TI - Macrolide antibiotics aggravate experimental autoimmune encephalomyelitis and inhibit inducible nitric oxide synthase. AB - Recent studies have implicated Chlamydia pneumoniae (C. pneumoniae) is present in a subset of patients with multiple sclerosis (MS) in which C. pneumoniae could act as a cofactor in the development of the disease. Macrolide antibiotics are most widely used anti-chlamydial agents and have immunomodulatory effect independently of their anti-bacterial activity. To investigate their effects on experimental autoimmune encephalomyelitis (EAE), EAE was induced by immunization with MBP68-86 peptide emulsified in complete Freund's adjuvant (CFA). Clarithromycin (CM) or azithromycin (AM, 50 mg/100 g body weight) was administrated daily from day 2 before immunization. All rats developed and survived EAE, but the groups administrated CM or AM had more severe symptoms. On day 11 post-immunization, mononuclear cells (MNCs) were prepared from the spleen of control group and cultured with or without macrolide antibiotics (10mug/ml). We evaluated nitric oxide (NO) production in the serum and culture supernatant. Inducible nitric oxide synthase (iNOS) mRNA and protein expression in the spinal cords and cultured MNCs were measured. The results showed that CM and AM similarly inhibited NO production and iNOS mRNA and protein expression in vivo and in vitro. Macrolide antibiotics may aggravate EAE by inhibiting iNOS mRNA and protein expression. Further studies are needed to investigate the effect of macrolide antibiotics on MS and to compare the effect of different anti chlamydial antibiotics on MS. PMID- 19811425 TI - Can mutational GC-pressure create new linear B-cell epitopes in herpes simplex virus type 1 glycoprotein B? AB - We showed that GC-content of nucleotide sequences coding for linear B-cell epitopes of herpes simplex virus type 1 (HSV1) glycoprotein B (gB) is higher than GC-content of sequences coding for epitope-free regions of this glycoprotein (G + C = 73 and 64%, respectively). Linear B-cell epitopes have been predicted in HSV1 gB by BepiPred algorithm ( www.cbs.dtu.dk/services/BepiPred ). Proline is an acrophilic amino acid residue (it is usually situated on the surface of protein globules, and so included in linear B-cell epitopes). Indeed, the level of proline is much higher in predicted epitopes of gB than in epitope-free regions (17.8% versus 1.8%). This amino acid is coded by GC-rich codons (CCX) that can be produced due to nucleotide substitutions caused by mutational GC-pressure. GC pressure will also lead to disappearance of acrophobic phenylalanine, isoleucine, methionine and tyrosine coded by GC-poor codons. Results of our "in-silico directed mutagenesis" showed that single nonsynonymous substitutions in AT to GC direction in two long epitope-free regions of gB will cause formation of new linear epitopes or elongation of previously existing epitopes flanking these regions in 25% of 539 possible cases. The calculations of GC-content and amino acid content have been performed by CodonChanges algorithm ( www.barkovsky.hotmail.ru ). PMID- 19811426 TI - Intrahepatic PD-1/PD-L1 up-regulation closely correlates with inflammation and virus replication in patients with chronic HBV infection. AB - Chronic hepatitis B was characterized by fluctuant immune response to infected hepatocytes resulting in hepatic inflammation and virus persistence. Recently, Programmed Death-1 (PD-1) and its ligand PD-L1 have been demonstrated to play an essential role in balancing antiviral immunity and inflammation in the livers of acute hepatitis B patients, significantly influencing disease outcome. PD-1 up regulation in peripheral T cells is associated with immune dysfunction in chronic hepatitis B patients. However, the effect of PD-1/PD-L1 on hepatic damage and chronic infective status is still unknown in patients with chronic HBV infection. Here, we report up-regulation of PD-1 and PD-L1 in liver biopsies from 32 chronic HBV patients compared to 4 healthy donors. PD-1/PD-L1 up-regulation was significantly associated with hepatic inflammation and ALT elevation. Moreover, appropriate up-regulation but not overexpression of PD-L1 in the active phase of chronic hepatitis B as well as lower expression of PD-L1 in the inactive phase in liver residential antigen presenting cells (including Kupffer cells and sinusoidal endothelial cells) may contribute to viral inhibition. Our data suggest that the intrahepatic interaction of PD-1 and PD-L1 might play an important role in balancing the immune response to HBV and immune-mediated liver damage in chronic HBV infection. PMID- 19811427 TI - Combined costimulation blockade inhibits accelerated rejection mediated by alloantigen-primed memory T cells in mice. AB - Donor-reactive memory T cells threaten the survival of transplanted organs via multiple pathways. This study was undertaken to induce tolerance of cardiac allografts in mice, in which alloreactive memory T cells were adoptively transferred, by combined costimulatory blockade of both effector and memory T cells. We found that the median survival time (MST) of the grafts was 5.17 days in the untreated group, 10.33 days in the CTLA4Ig- and anti-CD40L-treated (2 combined) group, and more than 100 days in the CTLA4Ig-, anti-CD40L-, anti-LFA-1 , and anti-OX40L-treated (4-combined) group. Histological analysis revealed that the mean rejection level was Grade 4 in the untreated group, Grade 3 in the 2 combined treatment group, and Grade 0 in the 4-combined treatment group. CD44(high) T cells were detected only in the untreated group. The in vitro proliferation of lymphocytes of both untreated and 2-combined group was higher than that of the 4-combined treatment group (p < 0.01). Compared with the untreated group, the expression levels of IL-2, IFN-gamma, and Foxp3 were lower in the 2-combined treatment group; the expression levels of these genes were the lowest in the 4-combined treatment group. IL-10 expression was significantly higher in the 4-combined treatment group than in the other groups. These results demonstrate the inhibition efficacy of combined costimulation blockade in accelerated-rejection models and the possible mechanisms underlying the suppression of cellular immunity in mice receiving grafts as well as in inducing the activation of IL-10-producing Tr1 cells in grafts. PMID- 19811428 TI - Activation of peripheral Th17 lymphocytes in patients with asthma. AB - A recently identified interleukin (IL)-17-producing T-helper (Th) lymphocyte subset, which comprises Th17 cells producing hallmark cytokines IL-17A, IL-17F and IL-22, is involved in chronic inflammatory diseases. Elevated gene and protein expressions of IL-17 are manifested in allergic asthma. We further characterized the activation of Th17 cells in asthmatic patients. Peripheral blood mononuclear cells (PBMC) were purified from 31 asthmatic patients and 20 sex- and age-matched control subjects. The number of IL-17A secreting cells in peripheral blood was enumerated by enzyme-linked immunosorbent spot assay. Cell surface expression of Th17-related chemokine receptor CCR6, and plasma level of IL-17A, IL-17F and IL-22, and ex vivo production of IL-17A and IL-22 were measured by flow cytometry and enzyme-linked immunosorbent assay, respectively. The number of peripheral Th17 lymphocytes, expression of CCR6 on Th cells, and ex vivo IL-23, anti-CD3 and anti-CD28 induced production of IL-22 by PBMC were significantly elevated in asthmatic patients compared with control subjects (all p < 0.01). This clinical study further confirmed increased number of peripheral Th17 lymphocytes and cell surface expression of CCR6 receptors on Th cells in asthmatic patients. Pro-inflammatory cytokine IL-23 can exacerbate disease severity by activating pathogenic Th17 lymphocytes to release downstream inflammatory cytokine IL-22 in asthma. PMID- 19811429 TI - An autologous therapeutic dendritic cell vaccine transfected with total lung carcinoma RNA stimulates cytotoxic T lymphocyte responses against non-small cell lung cancer. AB - OBJECTIVE: The development of immunotherapy for malignancy is greatly limited by the characteristic weak antigenicity of tumors. The primary goal of this study was to circumvent the isolation and purification of tumor-specific antigen determinants by producing a vaccine using lung tumor RNA-loaded dendritic cells (DCs), and to test the response against lung cancer. METHODS: Total RNA was isolated from 18 lung carcinomas with positive carcinoembryonic antigen (CEA) and mucin-1 (MUC1) staining, as identified by immunohistochemistry. DCs and T-cells from peripheral blood mononuclear cells were generated in vitro, and then DCs in different stages were transfected with RNA using several different methods. The expression of CEA and MUC1 in RNA-transfected DCs was measured using flow cytometry. T-cells stimulated by DCs were harvested as effectors, and primary tumor cells cultured in vitro were used as targets. Cytotoxicity was determined by lactic dehydrogenase detection assay. RESULTS: Immature RNA-transfected DCs significantly increased the expression of CEA and MUC1, compared to mature transfected DCs. RNA transfection via electroporation resulted in significantly greater CEA and MUC1 expression than did transfection via lipofection or passive pulsing. Lymphocytes stimulated by DCs transfected with lung tumor RNA initiated a cytotoxic T lymphocyte (CTL) tumor-specific response. CONCLUSION: Immature DCs transfected with total lung carcinoma RNA by electroporation in vitro effectively stimulate antigen-specific CTL responses against tumor cells. PMID- 19811430 TI - Immunological investigations. PMID- 19811431 TI - Genetic polymorphisms and immune responses. PMID- 19811432 TI - Interleukin 1 gene cluster, myocardial infarction at young age and inflammatory response of human mononuclear cells. AB - The objective was to investigate whether genotypes, haplotypes and haplotype pairs of interleukin (IL) gene cluster are associated with risk of Myocardial Infarction (MI) at young age and with the release of IL-1B and expression of tissue factor pro-coagulant activity (TFPCA), after stimulation in vitro with lipopolysaccharide (LPS) of human peripheral blood mononuclear cells (PBMCs). Patients with MI at young age, frequency-matched for age, sex and recruitment centre, with healthy population-based controls and PBMCs from healthy volunteers were studied. Five single nucleotide polymorphisms (SNPs), identifying two haplotype-blocks, in IL-1B gene and one SNP in IL-1A and IL-RA genes were genotyped. In multivariate analyses, haplotype A2 (122) and A4 (112) were associated with decreased risk of MI [OR = 0.62 (95% CI = 0.40-0.95), p = 0.01; OR = 0.69 (95% CI = 0.51-0.92), p = 0.03, respectively]. Haplotype-pair A2/A2 showed significant reduction in the risk of MI [OR = 0.38 (95% CI = 0.18-0.81); p = 0.01]. Haplotype A2 and A4 were associated with lower levels of IL-1B (respectively p = 0.01; p = 0.04, multivariate analysis) and haplotype-pair A2/A4 showed decreased levels of IL-1beta (p = 0.02). No association was found between block "B" IL-1B haplotypes or IL-1A and IL-RA polymorphisms and risk of MI. IL-1B haplotypes influence the inflammatory response of human mononuclear cells to LPS and affect the risk of MI at young age. PMID- 19811433 TI - Interleukin-1 receptor antagonist gene polymorphism and susceptibility to ischemic stroke. AB - Cytokines gene polymorphisms have been implicated in susceptibility to ischemic stroke. This study aims to determine the influence of the polymorphism within the intron 2 of the interleukin- 1 receptor antagonist (IL-1Ra) gene on the susceptibility to stroke. A variable number of tandem repeats (VNTR) in intron 2 of the IL-1Ra gene was analyzed in 148 patients with stroke and 161 healthy volunteers from the same area. The carriage rate of allele 2 of IL-1Ra gene, low producer, was significantly higher in patients with stroke compared to the controls (29% vs 21% p = 0.02). Frequency of IL1RN1/IL1RN1 genotype in the patients was significantly lower than the controls (49% vs 66% p = 0.003). The distribution of homozygous genotypes of IL1RN2 was not different between the controls and stroke patients while the heterozygous genotype was more frequent among the patients. (39% vs 25%, respectively). Multiple logistic regression analysis demonstrated that individuals who carry allele 2 for IL-1Ra gene had a significantly higher risk for ischemic stroke with an odds ratio of 2.48 (95% CI, 1.67, 3.51, p = 0.006). These data suggest that allele 2 of the IL-1Ra intron 2 gene represents a susceptibility factor in the development of ischemic stroke. PMID- 19811434 TI - Association between IL-1B and IL-1RN gene polymorphisms and Chagas' disease development susceptibility. AB - Though it is known that the immune system exerts some influence on the resistance against T. cruzi infection its precise role in this process is not well understood. Some IL-1B alleles and haplotypes have been associated with susceptibility to inflammatory, autoimmune and infectious diseases. The objective of this study was to determine and compare the distribution of IL-1B and IL-1 receptor antagonist (IL-1RN) polymorphisms among T. cruzi seropositive patients, patients with idiopathic dilated cardiomyopathy (IDC) and healthy individuals. We studied 86 individuals seropositive for T. cruzi (58 patients with chronic chagasic cardiomyopathy (CCC) and 28 asymptomatics), 50 seronegative individuals with IDC and 109 healthy individuals. IL-1B-511, IL-1F10.3 IL-1RN.4, IL-1RN 6/1, and IL-1RN 6/2 polymorphisms were analyzed using real-time PCR allelic discrimination technology. Infected patients presented an increased frequency of the CC genotype of the IL-1RN.4 polymorphism when compared to IDC (pC = 0.028; OR = 11.46). The C allele of this polymorphism was found increased in CCC when compared with IDC (pC = 0.036; OR = 0.5) and with controls (pC = 0.035; OR = 1.87). CC genotype of IL-1RN.4 polymorphism was increased in patients with CCC when compared to IDC (pC = 0.0018; OR = 16.74) and healthy individuals (pC = 0.011; OR = 3.6). There is an evident association between the IL1RN.4 polymorphism, T. cruzi infection and CCC development. PMID- 19811435 TI - Single nucleotide polymorphisms in TNF-alpha, TNFR2 gene and TNF-alpha production in Asian Indians. AB - Polymorphisms in the TNF-alpha and TNF receptor type 2 (TNFR2) genes in Asian Indians are not well understood. We investigated single nucleotide polymorphisms in the TNF-alpha 5'-flanking promoter/enhancer region and in exons 6, 9 and 10 of TNFR2 gene by PCR-restriction length polymorphism (PCR-RFLP) and PCR-sequence specific primer (SSP) techniques, respectively. The results showed single bi allelic polymorphism in TNF-alpha (-308G > A) and in TNFR2 exons 6 (676 T > G) and 9 (1176 G > A). Additionally, three bi-allelic polymorphisms (1663 G > A, 1668G > T and 1690C > T) were observed in the exon 10 of TNFR2 gene. The distribution of polymorphic alleles distinctly differed in Aryan and Dravidian Indian communities. The TNF-alpha and TNFR2 genotype and allele frequencies of Asian Indians stand closer to other Asian populations but distant from Caucasians. Furthermore, TNF-alpha -308 GA genotype was associated with significantly higher production of TNF-alpha as compared to GG genotype. These polymorphisms may be related to variation in TNF-alpha and TNFR2 expression during immune response to various stimuli in Asian Indians. PMID- 19811436 TI - The proinflammatory cytokine TNF-alpha -308 AA genotype is associated with polyglandular autoimmunity. AB - Data regarding polymorphisms of immunoregulatory genes in polyglandular autoimmunity (PGA) are lacking. We have analyzed whether the polymorphism of the proinflammatory cytokine gene TNF-alpha; -308 and mutations of the autoimmune regulator (AIRE) gene were associated with PGA in adults. Sixty-seven patients with PGA and 209 healthy controls were genotyped by multiplex minisequencing with capillary electrophoresis on an ABI PRISM-310 genetic analyzer. HLA DRB1 typing was performed using polymerase-chain-reaction-amplified DNA hybridized with sequence-specific-oligonucleotide probes (PCR-SSO). The TNF-alpha; -308*A allele occurred more frequently in patients (0.269) than in controls (0.163, P = 0.008, P(c) = 0.016). Also, TNF-alpha; -308*A carriers were more frequent in patients than controls (47.8% vs. 31.1%, OR = 1.89, 95%CI = 1.19-3.00). The frequency of the AA genotype was increased in PGA (P = 0.014, P(c) = 0.042). PGA patients with autoimmune thyroid disease and the TNF-alpha; -308 AA genotype showed the highest prevalence of thyroid autoantibodies (TPO, P = 0.04; Tg, P = 0.003). HLA-DRB1*03 and TNF-alpha; -308*A alleles were strongly associated in patients with PGA (87.5%, P(c) < 0.00001). The AIRE R257X and 13bpdel mutations were not observed in patients with PGA. The association of TNF-alpha; -308*A with PGA might be directly or indirectly due to the association with HLA-DRB1*03. PMID- 19811437 TI - Distribution of HLA-DRB1 alleles in Argentinean patients with Chagas' disease cardiomyopathy. AB - Chronic Chagas' disease occurs in a variable number of infected individuals and mainly manifests as an inflammatory cardiomyopathy that may lead to a fatal course. The factors underlying the establishment of chronic myocardial lesions are not fully understood. The study included 71 unrelated individuals serologically positive for T. cruzi. A group of 81 no related healthy individuals with neither symptoms nor previous diagnosis of Chagas' disease was studied as control group. Genomic DNA was extracted from peripheral blood using the standard salting out method and used as a template to amplify by the PCR the polymorphic second exon of the HLA-DRB1. PCR products were hybridized separately with sequence-specifics oligonucleotides (SSOP). DRB1*0409 and DRB1*1503 alleles were significantly more prevalent in seropositives (pC = 0.002, OR: 26.17 and 24.87 respectively). The prevalence of DRB1*1103 allele was statistically significant in the group control and could be associated with resistance Chagas' disease (pC = 0.026, OR: 0.19). Increased significance frequency of DRB1*1503 allele was found among cardiomyopathy patients suggesting that this antigen might be related with the genetic susceptibility to cardiac damage in these patients (pC = 0.014, OR: 9.22). PMID- 19811438 TI - HLA-DR allele frequencies in Mexican mestizos with autoimmune liver diseases including overlap syndromes. AB - Autoimmune liver diseases are sometimes difficult to differentiate from hepatic overlap syndromes (OS). The objective of this study was to use polymorphic genetic markers to better distinguish clinical heterogeneity in autoimmune liver disease. Since autoimmunity is the result of autoantibody production we studied HLA-DR alleles in 20 patients with autoimmune hepatitis (AIH), 16 with primary biliary cirrhosis (PBC), 10 with OS, and in 99 ethnically matched healthy individuals. Patients with OS had significantly higher alkaline phosphatase and total bilirubin levels than patients with AIH. OS patients had a higher prevalence of positive antinuclear antibodies and a higher AIH score than patients with PBC. Patients with OS also had higher total immunoglobulin levels (IgG isotype) as compared to patients with PBC. We found in PBC patients a higher gene frequency of HLA-DR4 and DR1 as compared to healthy controls (p = 0.03, OR = 2.2 and p = 0.004, OR = 4.3, respectively) and to OS patients (p = 0.01, OR = 6.8, and p = 0.004, OR = 10.0, respectively). On the other hand, the gene frequency of HLADR5 was significantly decreased in the total group of patients as compared to healthy controls suggesting a protective role of this allele for developing autoimmune liver disease. PMID- 19811439 TI - Effects of the CCR5-Delta32 mutation on hepatitis C virus-specific immune responses in patients with haemophilia. AB - In hepatitis C virus (HCV) infection antiviral T cells express the CC chemokine receptor 5 (CCR5). Their recruitment to the liver is an important step in the immune response. A 32 base pair deletion in the CCR5 gene leads to reduced expression and total loss of CCR5 in CCR5-Delta32 heterozygous and homozygous subjects, respectively. However, the role of this mutation for antiviral immunity remains unclear. Here, we analysed proliferation, IFN-gamma and IL-4 secretion (ELISpot) induced by the HCV antigens core, NS3, NS4, and NS5a in 21 anti-HCV positive haemophiliac patients in relationship to their CCR5 genotypes (CCR5 wildtype n = 10, CCR5-Delta32 heterozygous n = 5 and CCR5-Delta32 homozygous n = 6). Furthermore, T cell migration in response to the CCR5 ligands CCL3, -4 and -5 was studied. Overall IFN-gamma responses to HCV proteins were only slightly greater in CCR5 wild-type patients than in CCR5-Delta32 carriers (0.6 versus 0.24 SFC/10(4) PBMC; p = 0.043). This difference was consistently seen with all tested HCV antigens. In contrast, neither T cell migration, nor PBMC proliferation, nor IL-4 production differed between CCR5 genotypes. Interruption of the CCR5 signalling pathway due to CCR5-Delta32 may potentially result in subtle reduction of HCV specific IFN-gamma responses in anti-HCV-positive haemophiliac patients. PMID- 19811440 TI - TLR4 and IL-18 gene variants in chronic periodontitis: impact on disease susceptibility and severity. AB - The aim of the study was to assess whether genotypes in the Toll-like receptor 4 gene and in the promoter of the interleukin-18 gene are associated with the susceptibility to chronic periodontitis. 108 chronic periodontitis patients and 76 controls were genotyped for c.896A>G/1196C>T (TLR4 gene) and for c.-368G>C/ c. 838C>A (IL-18 promoter). There were no significant differences in genotype and allele distributions between the study groups. Periodontitis severity in patients with TLR4 c.896AG/1196CT genotype was significantly higher than wildtype carriers. The percentage of teeth with clinical attachment loss > or = 5 mm was 77.3% and 58.8%, respectively (p < or = 0.006, t-test). All subjects were further classified into carriers and non-carriers of at least one variant of each gene. A logistic regression analysis adjusted for gender, smoking, and age showed no association between gene variant carrier status and periodontitis (OR = 1.98, 95% CI 0.61-6.39). The results did not show that IL-18 and TLR4 variants have an effect on the susceptibility to chronic periodontitis. Considering the low number of periodontitis patients carrying TLR4 variants (11%), a comparison of the periodontitis severity depending on the genotype has to be interpreted cautiously. PMID- 19811441 TI - The functional effects of the -455G/A polymorphism on the IL-6-induced expression of the beta-fibrinogen gene may be due to linkage disequilibrium with other functional polymorphisms. AB - Elevated plasma fibrinogen levels have been identified as an independent risk factor for coronary heart diseases, stroke and peripheral artery disease. The 455G/A polymorphism in the promoter region of the beta-fibrinogen gene has been associated with increased plasma fibrinogen levels. However, the functional effect of this polymorphism has been controversial and other polymorphisms in the fibrinogen gene have also been implicated in higher fibrinogen levels. In this study, we evaluated the transcriptional activity of 4 natural haplotypes and 6 artificial haplotypes in the promoter region of the beta-fibrinogen gene. Significantly lower IL-6-induced activity was observed in the -1420A and -148T alleles. In contrast, the -854A allele had significantly higher activity. Artificial haplotypes containing the -1420A, -854A and -148T alleles were also analyzed to confirm individual functional effects. The -1420A and -148T alleles significantly lowered the activities, while the -854A allele significantly raised the activity. From this study we conclude that the -1420G/A, -854G/A and -148C/T polymorphisms in the beta-fibrinogen promoter region are functional polymorphisms while the -455G/A polymorphism may not be a functional one, and that the association of the -455G/A polymorphism with higher fibrinogen levels may actually be due to linkage disequilibrium between the -455G/A polymorphism and other truly functional polymorphisms. PMID- 19811442 TI - Immuno-modulatory gene polymorphisms and outcome in breast and ovarian cancer. AB - Breast and ovarian cancer continue to be a significant source of morbidity and mortality. Improved understanding of signalling pathways related to growth and apoptosis has led to targeted treatments and modest improvement in long term outcomes. However, it has become increasingly clear that tumor factors alone are not the sole determinants of outcome in patients with breast and ovarian cancer. The tumor microenvironment and other immunologic host processes play an integral role in the overall interactions between disease, host and treatment. Cytokines play a major role in the immune response to tumors. Single nucleotide polymorphisms (SNPs) in the regulatory or coding regions of many cytokine genes lead to functional alterations in the transcriptional regulation of these genes or the proteins they encode. This review examines the current literature linking functional variants in cytokine and other immune genes to outcomes in breast and ovarian cancer. We have focused on those involved in the proinflammatory response (IL-6, TNF-alpha), apoptosis (TGF-beta, Fas, FasL, C1QA), angiogenesis (IL-8) and autoimmunity (IL-10). While much remains to be learned about the mechanisms underlying these variants and their impact on tumor behavior, this area holds promise for future development of prognostic profiles and therapeutics exploiting the immune response. PMID- 19811443 TI - Transglutaminase-catalyzed crosslinking in neurological disease: from experimental evidence to therapeutic inhibition. AB - Diseases of polyglutamine expansion, Alzheimer's disease and Parkinson's disease are neurodegenerative diseases associated with insoluble protein aggregates and neuronal death. These diseases constitute a group of devastating diseases for which there is currently little treatment. The protein aggregates may be the cause of neuronal death, although there is some controversy as to which form of aggregation (oligomers, polymers or microscopic aggregates) is the most toxic. More than a decade ago, the participation of transglutaminases in the formation of the abnormal protein aggregates was proposed. Transglutaminases are a large family of enzymes that catalyze the formation of N(sigma) (gamma-glutamyl)-lysine isodipeptide crosslinks between proteins. In this review, we summarize the evidence supporting the participation of transglutaminase in diseases of the central nervous system. We also describe newly developed transglutaminase inhibitors and their potential use as therapeutic agents in neurological disease. PMID- 19811446 TI - Disease-modifying agents in the treatment of multiple sclerosis: a review of long term outcomes. AB - Multiple sclerosis (MS) is a chronic, debilitating condition mediated by inflammation and neurodegeneration. The ultimate goal of treatment is to delay or halt the progression of irreversible disability. Disease-modifying drugs (DMDs), including beta interferon and glatiramer acetate during phase III trails, have been shown to reduce relapse rates in relapsing-remitting multiple sclerosis (RRMS) as detected by magnetic resonance imaging (MRI). However, the long-term effects of DMDs on MS progression are not very clear; therefore, the aim of this paper is to evaluate the evidence available of the long-term effects of DMDs on reducing the progression of multiple sclerosis. A number of open-label, prospective extensions that followed a cohort of patients enrolled in double blind, placebo-controlled trials were examined. Methodological difficulties faced in designing a trial of extended duration were hard to overcome, however, and long-term, open-label extensions of interferon and glatiramer acetate failed to show significant beneficial effects in delaying disability progression, questioning the cost-effectiveness of these therapies in the long-term. PMID- 19811447 TI - The roles of corticotropin releasing factor (CRF) in responses to emotional stress: is CRF release a cause or result of fear/anxiety? AB - Corticotropin releasing factor (CRF) is a neuropeptide that is a major regulator of the hypothalamic-pituitary-adrenal system. Recent findings have shown that CRF exists in extrahypothalamic areas in the brain as well as in the hypothalamus, and extrahypothalamic CRF is also deeply involved in stress responses. Therefore, CRF has been a major target of drug development for treatment of stress-related disorders. However, whether CRF is a cause or a result of fear/anxiety has not been investigated extensively, even though this issue is extremely important to the development of treatments for stress-related disorders. This article aims to 1) introduce readers to several functional aspects of CRF, focusing on aspects that have been missed or ignored when determining the roles of CRF in responses to emotional stress; 2) critically review previous studies regarding the roles of CRF in responses to emotional stress, considering functional aspects of CRF described in 1); and 3) put forward a hypothesis about the roles of CRF in stress responses. Considering different functional aspects of CRF, it was suggested that CRF is a result of fear/anxiety, rather than a cause. In other words, CRF could be responsible for stress responses to cope with dangerous situations but not for fear/anxiety itself. CRF as a potential target of drug development for treatment of stress-related disorders is also discussed. PMID- 19811448 TI - No evidence of direct binding between ursodeoxycholic acid and the p53 DNA binding domain. AB - UDCA (ursodeoxycholic acid) is used increasingly for the treatment of cholestatic liver diseases. Among other cytoprotective effects, this endogenous bile acid is a potent inhibitor of apoptosis, interfering with both intrinsic and extrinsic apoptotic pathways. In previous studies, we have demonstrated that the transforming growth factor beta1-induced E2F-1/Mdm2 (murine double minute 2)/p53 apoptotic pathway was an upstream molecular target of UDCA. In agreement with this, we have recently established p53 as a key molecular target in UDCA prevention of cell death. The tumour suppressor p53 is a well-described transcription factor that induces the expression of multiple different pro apoptotic gene products. Its regulation involves a variety of signalling proteins and small molecules, and occurs at multiple levels, including transcription, translation and post-translation levels. In the present study, by using different biophysical techniques, we have investigated the possibility of a direct interaction between the p53 core domain, also referred to as the DNA-binding domain, and UDCA. Our in vitro analysis did not provide any evidence for direct binding between the bile acid UDCA and the p53 core domain. PMID- 19811449 TI - Cholesterol-dependent phase separation in cell-derived giant plasma-membrane vesicles. AB - Cell-derived GPMVs (giant plasma-membrane vesicles) enable investigation of lipid phase separation in a system with appropriate biological complexity under physiological conditions, and in the present study were used to investigate the cholesterol-dependence of domain formation and stability. The cholesterol level is directly related to the abundance of the liquid-ordered phase fraction, which is the majority phase in vesicles from untreated cells. Miscibility transition temperature depends on cholesterol and correlates strongly with the presence of detergent-insoluble membrane in cell lysates. Fluorescence correlation spectroscopy reveals two distinct diffusing populations in phase-separated cell membrane-derived vesicles whose diffusivities correspond well to diffusivities in both model systems and live cells. The results of the present study extend previous observations in purified lipid systems to the complex environment of the plasma membrane and provide insight into the effect of cholesterol on lipid phase separation and abundance. PMID- 19811450 TI - Clopidogrel, independent of the vascular P2Y12 receptor, improves arterial function in small mesenteric arteries from AngII-hypertensive rats. AB - The P2Y12 receptor antagonist clopidogrel blocks platelet aggregation, improves systemic endothelial nitric oxide bioavailability, and has anti-inflammatory effects. Since P2Y12 receptors have been identified in the vasculature, we hypothesized that clopidogrel ameliorates angiotensin II (Ang II) -induced vascular functional changes by blockade of P2Y12 receptors in the vasculature. Male Sprague Dawley rats were infused with Ang II (60 ng.min-1) or vehicle for 14 days. The animals were treated with clopidogrel (10mg*kg-1*day-1) or vehicle. Vascular reactivity was evaluated in second-order mesenteric arteries. Clopidogrel treatment did not change systolic blood pressure [(mmHg) control vehicle, 117+/-7.1 vs. control- Clopidogrel, 125+/-4.2; AngII-vehicle, 197+/-10.7 vs. AngII-Clopidogrel, 198+/-5.2], but it normalized increased phenylephrine induced vascular contractions [(%KCl) vehicle-treated, 182.2+/-18 vs. Clopidogrel, 133+/-14%), as well as impaired vasodilation to acetylcholine [(%) vehicle-treated, 71.7+/-2.2 vs. Clopidogrel, 85.3+/-2.8) in Ang II-treated animals. Vascular expression of P2Y12 receptor was determined by western blot. Pharmacological characterization of vascular P2Y12 was performed with the P2Y12 agonist 2-MeS-ADP. Although 2-MeSADP induced endothelium-dependent relaxation [(Emax %) = 71%+/-12), as well as contractile vascular responses (Emax %= 83+/ 12) these actions are not mediated by P2Y12 receptor activation. 2-MeS-ADP produced similar vascular responses in control and Ang II rats. These results indicate potential effects of Clopidogrel, such as improvement of hypertension related vascular functional changes that are not associated with direct actions of clopidogrel in the vasculature, supporting the concept that activated platelets contribute to endothelial dysfunction, possibly via impaired NO bioavailability. PMID- 19811451 TI - A putrescine-anthracene conjugate: a paradigm for selective drug delivery. AB - Increased polyamine concentrations play an important role in the development of cancer at all stages, from initiation through to maintenance of the transformed phenotype. One way cancer cells accumulate increased concentrations of polyamines is by increased uptake of preformed polyamines via their PTS (polyamine transport system). The PTS is promiscuous and will transport a range of polyamine-based molecules. Therefore it may be that cytotoxic drugs could be attached to polyamine vectors and targeted selectively to cancer cells by utilizing the PTS. The aim of the present study was to investigate the potential of Ant 4, a putrescine-anthracene conjugate, to target cytotoxic agents to human cancer cells as a paradigm for a novel method of selective drug delivery. Ant 4 induced cytotoxicity after only 24 h exposure. Apoptosis was the predominant type of cell death, with mechanistic studies revealing that oxidative stress and DNA damage may have a part to play. For the first time, uptake of Ant 4 via the PTS was demonstrated both directly and indirectly in human cell lines. In addition, Ant 4 significantly reduced putrescine uptake, demonstrating that this conjugate not only used the PTS, but also could successfully compete with its native polyamine for uptake. However, the most interesting finding was the intracellular depletion of the polyamine pools, providing an additional mode of toxicity for Ant 4 and the possibility that this molecule may act as a 'double-edged sword': preventing cell growth by delivery of the toxic moiety and by depletion of intracellular polyamine content. PMID- 19811452 TI - C/EBP transcription factors regulate SREBP1c gene expression during adipogenesis. AB - The transcription factor SREBP1c (sterol-regulatory-element-binding protein 1c) is highly expressed in adipose tissue and plays a central role in several aspects of adipocyte development including the induction of PPARgamma (peroxisome proliferator-activated receptor gamma), the generation of an endogenous PPARgamma ligand and the expression of several genes critical for lipid biosynthesis. Despite its significance, the regulation of SREBP1c expression during adipogenesis is not well characterized. We have noted that in several models of adipogenesis, SREBP1c expression closely mimics that of known C/EBPbeta (CCAAT/enhancer-binding protein beta) targets. Inhibition of C/EBP activity during adipogenesis by expressing either the dominant-negative C/EBPbeta LIP (liver-enriched inhibitory protein) isoform, the co-repressor ETO (eight-twenty one/MTG8) or using siRNAs (small interfering RNAs) targeting either C/EBPbeta or C/EBPdelta significantly impaired early SREBP1c induction. Furthermore, ChIP (chromatin immunoprecipitation) assays identified specific sequences in the SREBP1c promoter to which C/EBPbeta and C/EBPdelta bind in intact cells, demonstrating that these factors may directly regulate SREBP1c expression. Using cells in which C/EBPalpha expression is inhibited using shRNA (short hairpin RNA) and ChIP assays we show that C/EBPalpha replaces C/EBPbeta and C/EBPdelta as a regulator of SREBP1c expression in maturing adipocytes. These results provide novel insight into the induction of SREBP1c expression during adipogenesis. Moreover, the findings of the present study identify an important additional mechanism via which the C/EBP transcription factors may control a network of gene expression regulating adipogenesis, lipogenesis and insulin sensitivity. PMID- 19811453 TI - A di-arginine motif contributes to the ER localization of the type I transmembrane ER oxidoreductase TMX4. AB - The thiol-disulfide oxidoreductases of the PDI (protein disulfide isomerase) family assist in disulfide-bond formation in the ER (endoplasmic reticulum). In the present study, we have shown that the previously uncharacterized PDI family member TMX4 (thioredoxin-like transmembrane 4) is an N-glycosylated type I membrane protein that localizes to the ER. We also demonstrate that TMX4 contains a single ER-luminal thioredoxin-like domain, which, in contrast with similar domains in other PDIs, is mainly oxidized in living cells. The TMX4 transcript displays a wide tissue distribution, and is strongly expressed in melanoma cells. Unlike many type I membrane proteins, TMX4 lacks a typical C-terminal di-lysine retrieval signal. Instead, the cytoplasmic tail has a conserved di-arginine motif of the RXR type. We show that mutation of the RQR sequence in TMX4 to KQK interferes with ER localization of the protein. Moreover, whereas the cytoplasmic region of TMX4 confers ER localization to a reporter protein, the KQK mutant of the same protein redistributes to the cell surface. Overall, features not commonly found in other PDIs characterize TMX4 and suggest unique functional properties of the protein. PMID- 19811455 TI - Helmet-delivered continuous positive airway pressure with heliox in respiratory syncytial virus bronchiolitis. AB - AIM: The objective of this study was to check the feasibility and efficacy of helmet-delivered heliox-continuous positive airway pressure (CPAP) in infants with bronchiolitis. METHODS: Children <3 months of age diagnosed with respiratory syncytial virus bronchiolitis and recurrent apnoeas or a venous PCO(2) >55 mmHg or a transcutaneous oxygen saturation <92% in room air were eligible for inclusion in the study. CPAP was delivered by a noninvasive ventilator connected to a heliox port. The interface was a helmet. RESULTS: Eight consecutive infants fulfilled the inclusion criteria. Apnoeas were present in six children before respiratory support was started; they disappeared in five of them. Two infants had to be changed to pressure support noninvasive ventilation, and one of them required intubation. No side effects were recorded. CONCLUSION: We propose a relatively new device to deliver heliox-CPAP in small infants with bronchiolitis. Although this is just a descriptive study with a short sample, this system seems to be feasible and effective. PMID- 19811456 TI - Chronic fatigue could be a marker of ADHD in children and adolescents. PMID- 19811457 TI - Longitudinal correlates of the persistence of irregular eating from age 5 to 14 years. AB - AIM: To report the stability of parent-perceived child irregular eating from 6 months to 14 years of age and to investigate a predictive model inclusive of child and parent factors. METHODS: Of the 7223 singleton children in a birth cohort, 5122 children were re-interviewed at 5 years and 4554 for the 14-year analysis. Information was obtained from structured interviews including questions answered by parents of the child at birth, 6 months, 5 years and 14 years; and by teenagers at age 14 years and from physical measures of the child. The mother's perception that the child was an irregular eater at age 14 years was the major outcome variable of interest. RESULTS: Approximately 40% of irregular eaters at age 5 will still be irregular eaters at age 14 years. This was not related to maternal education or socio-economic class. Significant at multivariate analysis were infant feeding problems and the children's ability to regulate their sleep and mood. Significant maternal factors were greater age, not feeling positive about the baby and persistent maternal anxiety during the child's early years. CONCLUSION: Irregular eating behaviour displays considerable continuity from childhood to mid-adolescence. Independent contributions to this behavioural phenotype include child biological and psychological factors and maternal anxiety during the child's early years. PMID- 19811458 TI - Enuresis in hyperthyroidism: a temporary lack of central control mechanism leads to nocturnal enuresis. AB - We report on a 9-year-old boy who suffered from hyperthyroidism and a new appearance of enuresis. Bedwetting ceased and prepulse inhibition (PPI) - measured as a parameter of central control - increased during the course of therapy. CONCLUSION: The increase in PPI is an indication that enuresis in hyperthyroidism could be as a result of a temporary loss of central control on brainstem reflexes. The case conveys new insights into the correlation between thyroid hormones and micturition patterns and the aetiology of enuresis. PMID- 19811459 TI - Transcutaneous bilirubinometry reduces the need for blood sampling in neonates with visible jaundice. AB - OBJECTIVES: We determined usefulness of transcutaneous bilirubinometry to decrease the need for blood sampling to assay serum total bilirubin (STB) in the management of jaundiced healthy Indian neonates. METHODS: Newborns, > or =35 weeks' gestation, with clinical evidence of jaundice were enrolled in an institutional approved randomized clinical trial. The severity of hyperbilirubinaemia was determined by two non-invasive methods: i) protocol-based visual assessment of bilirubin (VaB) and ii) transcutaneous bilirubin (TcB) determination (BiliCheck). By a random allocation, either method was used to decide the need for blood sampling, which was defined to be present if assessed STB by allocated method exceeded 80% of hour-specific threshold values for phototherapy (2004 AAP Guidelines). RESULTS: A total of 617 neonates were randomized to either TcB (n = 314) or VaB (n = 303) groups with comparable gestation, birth weight and postnatal age. Need for blood sampling to assay STB was 34% lower (95% CI: 10% to 51%) in the TcB group compared with VaB group (17.5% vs 26.4% assessments; risk difference: -8.9%, 95% CI: -2.4% to -15.4%; p = 0.008). CONCLUSION: Routine use of transcutaneous bilirubinometry compared with systematic visual assessment of bilirubin significantly reduced the need for blood sampling to assay STB in jaundiced term and late-preterm neonates. (ClinicalTrials.gov number, NCT00653874). PMID- 19811461 TI - The role of cytokine expression in different subgroups of subfertile men. AB - PROBLEM: The aim of this study was to evaluate the levels of seminal plasma cytokines interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 10 (IL-10), interleukin 11 (IL-11), interleukin 12 (IL-12), tumour necrosis factor alpha (TNF alpha) and interferon gamma (IFN-gamma) in male subfertility. METHOD OF STUDY: A total of 73 male partners of an infertile couple attending a regional andrology unit were recruited into this prospective study and subdivided into the various groups based on semen analysis. Concentrations of cytokines such as IL-6, IL-8, IL-10, IL-11, IL-12, TNF-alpha and IFN-gamma in the seminal plasma were determined using enzyme linked immunosorbent assay (ELISA). RESULTS: Significant higher concentrations (P < 0.05) of IL-6 in the mild and severe oligospermic group, IL-8 and IL-10 in the asthenospermic group and IL-6, IL-10, TNF-alpha and IFN-gamma in the obstructed azoospermic group were determined. IL-10 concentrations correlated significantly with other cytokines in the obstructed azoospermic group and the asthenospermic group. CONCLUSION: Our study confirms that cytokines rarely act in isolation, but rather in a network of other cytokines and may affect sperm function directly or indirectly. The presence of increased levels of cytokines in the obstructed azoospermic group suggests that the cytokines may not originate from the testis. PMID- 19811460 TI - A role for the volume regulated anion channel in volume regulation in the murine CNS cell line, CAD. AB - AIM: The role of the volume regulated anion channel (VRAC) in a model CNS neuronal cell line, CAD, was investigated. METHODS: Changes in cell volume following hypotonic challenges were measured using a video-imaging technique. The effect of the Cl(-) channel antagonists tamoxifen (10 microm) and 4,4' diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS; 100 microm) on regulatory volume decrease (RVD) were measured. The whole-cell voltage-clamp technique was used to characterize ICl(swell), the current underlying the VRAC. RESULTS: Using the video-imaging technique, CAD cells were found to swell and subsequently exhibit RVD when subjected to a sustained hypotonic challenge from 300 mOsmol kg( 1) H(2)O to 210 mOsmol kg(-1) H(2)O. In the presence of tamoxifen (10 microm) or DIDS (100 microm) RVD was abolished, suggesting a role for the VRAC. A hypotonic solution (230 mOsmol kg(-1) H(2)O) evoked ICl(swell), an outwardly rectifying current displaying time-independent activation, which reversed upon return to isotonic conditions. The reversal potential (E(rev)) for ICl(swell) was -14.7 + or - 1.4 mV, similar to the theoretical E(rev) for a selective Cl(-) conductance. ICl(swell) was inhibited in the presence of DIDS (100 microm) and tamoxifen (10 microm), the DIDS inhibition being voltage dependent. CONCLUSIONS: Osmotic swelling elicits an outwardly rectifying Cl(-) conductance in CAD cells. The ICl(swell) observed in these cells is similar to that observed in other cells, and is likely to provide a pathway for the loss of Cl(-) which leads to water loss and RVD. As ischaemia, brain trauma, hypoxia and other brain pathologies can cause cell swelling, CAD cells represent a model cell line for the study of neuronal cell volume regulation. PMID- 19811462 TI - PD-1 but not CTLA-4 blockage abrogates the protective effect of regulatory T cells in a pregnancy murine model. AB - PROBLEM: Regulatory T cells (Treg) play an important role in fetal protection. They expand during normal pregnancy and protect paternal/fetal antigens from rejection by maternal effector cells. Accordingly, the transfer of Treg obtained from BALB/c-mated CBA/J females prevents abortion in DBA/2J-mated animals. The actual mechanism through which Treg mediate their protective effect is still inconclusive. Cytotoxic T lymphocyte antigen-4 (CTLA-4) and Programmed cell death 1 (PD-1) are some of known Treg-associated molecules; however, their role in Treg mediated fetal protection in murine model has not been investigated. METHOD OF STUDY: Treg obtained from normal pregnant animals (NP; CBA/J x BALB/c) on day 14 were adoptively transferred into abortion-prone mice (AP; CBA/J x DBA/2J) intravenously on day 2 of pregnancy. An amount of 250 microg of either anti-PD-1 or anti-CTLA-4 mAb were injected intraperitoneally on days 0, 3, 6 and 9 of pregnancy. Controls received Treg + IgG or Treg + PBS. NP or AP treated with PBS served as additional controls. RESULTS: Blocking PD-1 abrogated the protective effect of Treg, resulting in a higher median abortion rate in comparison with the Treg/isotype-treated control while CTLA-4 blockage did not interfere with the protective effect of Treg. This was associated with a diminished number of vascular endothelial growth factor-A(+) cells, previously reported as stimulators of lymphocyte extravasation in preterm labor. CONCLUSION: Our data suggest PD-1 as an important mediator in Treg-induced fetal protection in the CBA/J x DBA/2J murine model. PMID- 19811463 TI - Pre-eclampsia, placental abruption and increased risk of atopic sensitization in male adolescent offspring. AB - PROBLEM: The aim of this study was to investigate maternal pre-eclampsia (PE), placental abruption (PA) and atopy among offspring. METHOD OF STUDY: Obstetric data was recorded from 378 women who were included into the Caesarean delivery birth cohort during 1990-1992. When their children were 15-17 years old skin prick tests (SPTs) and inhalant allergen-specific (IAS) immunoglobulin E (IgE) were determined among offspring. RESULTS: Adolescents whose mothers had either PE or PA at the time of delivery had more atopy and especially more severe atopy as measured by the incidence of SPTs positive for > or = 5 allergens (RR: 4.28, 95% CI: 1.54-11.92; P < 0.005) and elevated IAS IgE levels (> or = 1.0 kU/L) (RR: 4.27, 95% CI: 1.08-16.99; P < 0.039). This increased risk was particularly observed in male adolescents. CONCLUSION: Maternal PE and PA were associated with an increased risk of severe atopy, especially in male adolescent offspring. PMID- 19811464 TI - A high dose of intravenous immunoglobulin increases CD94 expression on natural killer cells in women with recurrent spontaneous abortion. AB - PROBLEM: A high dose of intravenous immunoglobulin (HIVIg) therapy is effective in various diseases such as autoimmune diseases, and also is expected to have efficacy in recurrent spontaneous abortion (RSA). The aim of this study was to understand immunological mechanisms of this therapy. METHOD OF STUDY: By flowcytometric analyses, we examined phenotypic changes of a variety of immunological cells including natural killer (NK) cells, cytotoxic T cells, regulatory T cells and macrophages in peripheral blood of RSA women with HIVIg therapy (n = 8). RESULTS: Expression percentages of inhibitory CD94 on NK cells significantly (P = 0.01) increased after the therapy (58.8 +/- 21.4% versus 71.0 +/- 17.6%). CONCLUSION: Mechanisms of possible efficacy of HIVIg therapy for RSA may include enhancement of CD94 expression and subsequent suppression of NK cell cytotoxicity. PMID- 19811465 TI - An interleukin-23 binding protein in mid-trimester amniotic fluid. AB - PROBLEM: The binding of mid-trimester amniotic fluid to cytokines was evaluated. METHOD OF STUDY: Purified tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-10, IL-12, and IL-23 were incubated with amniotic fluid from 25 women undergoing a mid-trimester amniocentesis, or with bovine serum albumin or saline, and cytokine binding to monoclonal antibodies was quantitated by ELISA. Aliquots of amniotic fluid were heated to 95 degrees C for 15 min and then retested for IL 23 binding. The effect of amniotic fluid dilution on IL-23 quantitation was evaluated. RESULTS: All amniotic fluids had a negligible effect on TNF-alpha, IL 10, and IL-12 detection. In marked contrast, pre-incubation with amniotic fluid from each subject reduced the subsequent ability to detect IL-23 by >50%. The extent of inhibition was directly proportional to the amniotic fluid dilution and was markedly reduced following heating at 95 degrees C for 15 min. Amniotic fluids from White, Black, Asian, East Indian, and Hispanic women were equally effective. CONCLUSION: Interleukin-23 and IL-12 share a common p40 subunit and no inhibition of IL-12 was apparent. It appeared that a component of mid-trimester amniotic fluid specifically interacts with the p19 subunit unique to IL-23. Mid trimester amniotic fluid reactivity with IL-23 may be a mechanism to limit intra amniotic neutrophil-derived inflammation. PMID- 19811466 TI - Live birth rate according to maternal age and previous number of recurrent miscarriages. AB - PROBLEM: In Japan, marital age and women's age at the first pregnancy are continuing to increase year by year. However, information concerning subsequent live birth rate according to maternal age and number of previous recurrent miscarriages is limited. METHOD OF STUDY: We studied a total of 1250 unexplained patients suffering two or more consecutive miscarriages. We examined the live birth rate at the first pregnancy and the cumulative success rate for birth of at least one child after examination. RESULTS: The live birth rate of women in their 40s was 58.1%, which was similar to that of women who were 35-39 years old (58.4%) at the first pregnancy, as found after examination. From logistic regression, women's age and the number of previous miscarriages independently decreased the live birth rate in subsequent pregnancies (p(s)) as well as cumulative pregnancies (p(c)), as follows: logit (p(s)) = 3.964 - 0.0652 x (age) 0.408 x (previous number of miscarriages), logit(p(c)) = 6.806 - 0.1130 x (age) 0.514 x (previous number of miscarriages). CONCLUSION: The information concerning the live birth rate can be given to each patient before subsequent pregnancy. PMID- 19811467 TI - Soluble human leukocyte antigen-G isoforms in maternal plasma in early and late pregnancy. AB - PROBLEM: Human Leukocyte Antigen (HLA)-G is a class Ib gene located in the human major histocompatibility complex (MHC). Several lines of investigation indicate that the HLA-G molecule is involved in the maternal acceptance of the semi allogenic fetus during pregnancy and in the development of tolerance. Expression of soluble HLA-G (sHLA-G) is positively correlated with successful in vitro fertilization (IVF) treatments, and aberrant expression of HLA-G in certain complications of pregnancy, such as pre-eclampsia and spontaneous abortion, has been reported. The main purpose of this study was to investigate the levels of different soluble HLA-G isoforms in maternal plasma in early and late pregnancy. METHOD OF STUDY: Soluble HLA-G (sHLA-G) can be detected in maternal blood, and in this study, two different isoforms of sHLA-G, namely sHLA-G1 generated by shedding of membrane-bound HLA-G1 and HLA-G generated by specific HLA-G transcripts, have been investigated early [median of 16.4 weeks of gestation (GW)] and late (median: 38.9 GW) in pregnancy in an original cohort of 580 pregnant Caucasian women. RESULTS: Lower concentrations of sHLA-G1 were found late in pregnancy (>32 GW) in a group of women with severe pre-eclampsia compared with controls with uncomplicated pregnancies (P = 0.029, P(C) = 0.09; Mann Whitney; Logistic regression analysis: P = 0.024, OR = 0.920, 95% CI: 0.855 0.989). However, this was not the case with HLA-G5, and significantly more of the cases with severe pre-eclampsia had detectable plasma HLA-G5 compared with that of the control group (P = 0.013, P(C) = 0.04; Mann-Whitney). Similar findings were not observed in women with gestational hypertension or existing hypertension continuing into pregnancy. Furthermore, there was a trend toward lower maternal plasma sHLA-G1 in a group of women with premature birth (<37 GW) compared with that of the control group (P = 0.028, P(C) = 0.17; Mann-Whitney). On the contrary, HLA-G5 was lower in the control group compared with that in the premature group (P = 0.004, P(C) = 0.02; Mann-Whitney). CONCLUSION: This study shows in line with other published studies that a high, detectable soluble HLA-G concentration in maternal plasma or serum is not mandatory for a successful pregnancy. However, complications during pregnancy, such as (severe) pre eclampsia, spontaneous abortion, IUGR, and premature birth, are associated with a low or undetectable level of soluble HLA-G in the maternal blood circulation. Also, this study indicates that sHLA-G1 is the interesting soluble HLA-G isoform in pre-eclampsia, and that low or undetectable levels of HLA-G5 at the end of pregnancy seem to be associated with an uncomplicated normal pregnancy, whereas in severe pre-eclampsia and possibly other pregnancy complications, such as preterm birth and IUGR, the level of HLA-G5 is higher. PMID- 19811468 TI - Multiple cytokine profile in plasma and amniotic fluid in a mouse model of pre term labor. AB - PROBLEM: The rate of pre-term birth in the United States continues to rise despite several interventions. Induction of pro-inflammatory cytokines and chemokines has been implicated in the activation of the cascade of events resulting in pre-term labor. To date, no comprehensive panel of the cytokine profile in PTL has been published. METHOD OF STUDY: To address cytokine profiles in pre-term labor, levels of 19 plasma and amniotic fluid cytokines were measured using a multiplex immunoassay in an inflammation-induced murine model of pre-term labor. RESULTS: Pro-inflammatory mediators, RANTES, KC, IL-6, and IL-12p40 were increased by 3 hr and remained high at 15 hr. Concentrations of KC, IL-6, IL 1beta, and MIP-1alpha were increased in the amniotic fluid at 15 hr. Plasma levels of anti-inflammatory mediators IL-10 and IL-13 at 15 hr were unchanged and decreased respectively. CONCLUSION: These results suggest that stimulation of several pro-inflammatory cytokines occurs very early in the cascade of events and remains increased, whereas anti-inflammatory cytokines are either unchanged or decreased until the onset of delivery in an inflammation-induced mouse model of pre-term labor. PMID- 19811470 TI - Abstracts of Future Challenges in Fertility Treatment, the Fertility Society of Australia. October 25-28, 2009. Perth, Australia. PMID- 19811472 TI - Are we really committed to making a difference? Reflections on Indigenous health research and dissemination. PMID- 19811473 TI - Qualitative method and the curse the illustrative quotation. PMID- 19811474 TI - Was it good for you too? Impediments to conducting university-based collaborative research with communities experiencing disadvantage. AB - OBJECTIVE: Collaborative and participatory research (CPR) models are increasingly recognised as methodologically, ethically and practically appropriate to conducting health and welfare research involving disadvantaged communities. This paper identifies impediments to CPR and proposes measures to support and encourage future CPR in Australian universities. METHODS: This paper draws on a small qualitative study of university-based CPR projects in Melbourne. The study involved a literature review and interviews with 23 participants, comprising university-based researchers and community liaison officers, and community representatives involved in university-based research projects. RESULTS: The paper outlines four main difficulties encountered by university-based researchers and community liaison staff in conducting CPR. These are: managing community sensitivities, the time-consuming nature of the work and diverse tasks involved, difficulty securing adequate research funding, and a concern that CPR was detrimental to academic careers. CONCLUSION: CPR in universities might be supported in the future through providing CPR training for researchers, employing additional community liaison staff, recognising community reports within the Australian research quality evaluation system Excellence in Research for Australia, adopting supportive policies within universities and provision of dedicated CPR funding. IMPLICATIONS: In the current Australian university context of competitive funding, further research into CPR nationally, alongside dedicated resources and policies are required to maximise the benefits of this approach. PMID- 19811475 TI - Baseline indicators for measuring progress in preventing falls injury in older people. AB - OBJECTIVE: Over recent years, there has been increasing attention given to preventing falls and falls injury in older people through policy and other initiatives. This paper presents a baseline set of fall injury outcome indicators against which these preventive efforts can be assessed in terms of monitoring the rate of fall-related deaths and hospitalisations. METHODS: ICD-10-AM coded hospital separations, Australian Bureau of Statistics (ABS) mortality and ABS population data were used to determine the rate of fall-related injury mortality and hospitalisations occurring in people aged 65+ years in New South Wales (NSW), Australia, over the six-year period from 1998/99 to 2003/04, inclusive. RESULTS: Baseline trends for one fatality and five separations-based metrics are presented. Overall, fall mortality rates increased over the six years, with higher rates in males. Falls hospitalisation rates also increased slightly, with higher rates in females. The rates of hip fracture and pelvic fracture hospital separations generally declined over the six years and were highest in females. The level of unspecified and missing information about the place where falls occur increased by 1.5%. CONCLUSION: Baseline trends in fall injury outcome metrics highlight the severity and frequency of fall injuries before wide scale implementation of the Management Policy to Reduce Fall Injury Among Older People in NSW. IMPLICATIONS: Future use of these metrics will help to evaluate and monitor the progress of falls prevention in older people in NSW. They could also be adopted in other jurisdictions. PMID- 19811476 TI - A new index of access to primary care services in rural areas. AB - OBJECTIVE: To outline a new index of access to primary care services in rural areas that has been specifically designed to overcome weaknesses of using existing geographical classifications. METHODS: Access was measured by four key dimensions of availability, proximity, health needs and mobility. Population data were obtained through the national census and primary care service data were obtained through the Medical Directory of Australia. All data were calculated at the smallest feasible geographical unit (collection districts). The index of access was measured using a modified two-step floating catchment area (2SFCA) method, which incorporates two necessary additional spatial functions (distance decay and capping) and two additional non-spatial dimensions (health needs and mobility). RESULTS: An improved index of access, specifically designed to better capture access to primary care in rural areas, is achieved. These improvements come from: 1) incorporation of actual health service data in the index; 2) methodological improvements to existing access measures, which enable both proximity to be differentiated within catchments and the use of varying catchment sizes; and 3) improved sensitivity to small-area variations. CONCLUSION: Despite their recognised weaknesses, the Australian government uses broad geographical classifications as proxy measures of access to underpin significant rural health funding programs. This new index of access could provide a more equitable means for resource allocation. IMPLICATIONS: Significant government funding, aimed at improving health service access inequities in rural areas, could be better targeted by underpinning programs with our improved access measure. PMID- 19811477 TI - The capacity of dental therapists to provide direct restorative care to adults. AB - INTRODUCTION: In Victoria, dental therapists are restricted to treating patients under the age of 26 years. Removing this age restriction from dental therapists' scope of practice may assist significantly in addressing workforce shortages, particularly in rural Victoria. OBJECTIVES: This study aims to assess the capacity of dental therapists to provide direct coronal restorations (dental fillings) to patients older than 25 years, on the prescription of a dentist. Its objectives include determining the success rate of restorations placed by dental therapists six months post placement; and patients' and dental therapists' satisfaction with the services provided. METHODS: The project was carried out in 2007 at the Royal Dental Hospital of Melbourne. Seven dental therapists participated in the study, placed 356 restorations (115 patients) with the support of a dentist. These restorations were reviewed six-months post placement by dentists blinded as to which restorations were placed by the dental therapists. Patients' age ranged from 26 to 82 years (82% were >40 years). RESULTS: At six months post-treatment, 258 restorations (80 patients) were reviewed. At review, 94.6% of the restorations were successful. Patients and dental therapists were satisfied with the experience. CONCLUSIONS: The standard of restorations provided by dental therapists was considered to be at least similar to that expected of a newly graduated dentist. IMPLICATIONS: Broadening the dental therapists scope of practice would create opportunities to design more flexible 'oral health' clinical teams enabling dentists to provide more complex procedures for patients most in need. This is significant in the public sector and rural areas where workforce shortages are most acute. PMID- 19811478 TI - Cost and cost-effectiveness of digital mammography compared with film-screen mammography in Australia. AB - OBJECTIVE: A systematic review assessed the relative safety and effectiveness of digital mammography compared with film-screen mammography. This study utilised the evidence from the review to examine the economic value of digital compared with film-screen mammography in Australia. METHODS: A cost-comparison analysis between the two technologies was conducted for the overall population for the purposes of breast cancer screening and diagnosis. In addition, a cost effectiveness analysis was conducted for the screening subgroups where digital mammography was considered to be more accurate than film-screen mammography. RESULTS: Digital mammography in a screening setting is $11 more per examination than film-screen mammography, and $36 or $33 more per examination in a diagnostic setting when either digital radiography or computed radiography is used. In both the screening and diagnostic settings, the throughput of the mammography system had the most significant impact on decreasing the incremental cost/examination/year of digital mammography. CONCLUSION: Digital mammography is more expensive than film-screen mammography. Whether digital mammography represents good value for money depends on the eventual life-years and quality adjusted life-years gained from the early cancer diagnosis. IMPLICATIONS: The evidence generated from this study has informed the allocation of public resources for the screening and diagnosis of breast cancer in Australia. PMID- 19811479 TI - Prevalence and impact of musculoskeletal disorders in New Zealand nurses, postal workers and office workers. AB - OBJECTIVES: To describe the prevalence, characteristics and impact of musculoskeletal disorders (MSDs) in New Zealand nurses, postal workers and office workers. METHODS: A postal survey asked participants about MSDs, (low back, neck, shoulder, elbow, wrist/hand or knee pain lasting longer than one day), and demographic, physical and psychosocial factors. Nurses were randomly selected from the Nursing Council database, postal workers from their employer's database and office workers from the 2005 electoral roll. RESULTS: The response rate of potentially eligible participants was 58% (n=443). Participants were aged 20-59 years; 86% were female. Over the 12 months prior to the survey 88% of respondents had at least one MSD lasting longer than a day and 72% reported an MSD present for at least seven days. Of the 1,003 MSDs reported, 18% required time off work and 24% required modified work duties. In the month prior to the survey 17% of MSDs made functional tasks difficult or impossible. Low back, neck and shoulder pain prevalence did not differ by occupation. Postal workers had the highest prevalence of elbow and wrist/hand pain; nurses of knee pain. CONCLUSIONS: The high prevalence of MSDs among these workers indicates that they are indeed in 'at risk' occupations. In each occupational group MSDs encompass a range of anatomical sites, however the overall pattern of MSDs differs by occupation. MSDs have a significant impact on activities at work and home. IMPLICATIONS: Primary and secondary prevention strategies should encompass a range of anatomical sites and specifically target different occupational groups. PMID- 19811480 TI - Hepatitis C virus infection rates and risk factors in an Australian hospital endoscopy cohort. AB - OBJECTIVE: To determine the reservoir and risk factors of HCV infection in a hospital population. METHODS: The presence of anti-HCV in 2,119 endoscopy patients was related to putative risk factors for exposure using the SAS statistical package. RESULTS: Most of the 4.7% of anti-HCV positive patients had multiple risk factors for HCV exposure. The risk was significantly increased in patients; with a previous history of hepatitis (36.4 fold), past history of injecting drugs (IDU) (32.1 fold), those born in North Africa, Middle East and Mediterranean countries (4.3 fold), had been tattooed before 1980s (3.3 fold), from 1980s-1990s (5.9 fold), had acupuncture before 1980s (3.8 fold), had a blood transfusion (3.6 fold), had clotting factors or growth hormone (4 fold), had contact with someone diagnosed with hepatitis in 1990s (4.1 fold). Of the anti HCV patients 38 had a history of IDU, 43 were migrants and 10 were both. CONCLUSION: Anti-HCV prevalence was five times higher than predicted by the passive surveillance scheme and 20% of patients were unaware of their infection. Only one of these patients reported IDU. The evidence of HCV intersecting epidemics between developing and developed countries in Australia was strongly supported. IMPLICATIONS: The study provides a rational basis for targeted programs to identify asymptomatic HCV carriers who might benefit from the new antiviral treatment. PMID- 19811481 TI - Absence of significant dissent should be sufficient for deceased donor organ procurement in New Zealand. AB - OBJECTIVE: New Zealand's organ donation rates are among the lowest in the OECD. In a bid to increase organ availability, the New Zealand Human Tissue Act 2008 introduces new consent arrangements for deceased donor organ procurement. This article assesses these new arrangements and presents the case for further reform. APPROACH: Our assessment and arguments are based on philosophical analysis informed by empirical data on the effectiveness of alternative consent systems. We: 1) Identify widely held ethical judgments about policies and practices relevant to organ donation (e.g. those relating to coronial post-mortems), 2) Assess the implications of these judgments for the Human Tissue Act and the assumptions that underpin it, and 3) Derive policy recommendations that are consistent with the judgments. CONCLUSION: The Human Tissue Act 2008 retains a strong consent requirement for organ procurement: organs may not be transplanted unless either the deceased or the family consents. We argue that organ availability could and should be increased by shifting from a model that requires consent to one that requires the absence of significant dissent. IMPLICATIONS: We recommend that New Zealand adopt either 1) an organ donation system similar to the existing system for ordering coronial post-mortems, or 2) a variant of the 'opt-out' system already in place in several other countries. PMID- 19811482 TI - Commentary: Absence of significant dissent should be sufficient for deceased organ procurement in New Zealand. PMID- 19811484 TI - Is inequity undermining Australia's 'universal' health care system? Socio economic inequalities in the use of specialist medical and non-medical ambulatory health care. AB - OBJECTIVES: To quantify need-adjusted socio-economic inequalities in medical and non-medical ambulatory health care in Australia and to examine the effects of specific interventions, namely concession cards and private health insurance (PHI), on equity. METHODS: We used data from a 2004 survey of 10,905 Australian women aged 53 to 58 years. We modelled the association between socio-economic status and health service use--GPs, specialists, hospital doctors, allied and alternative health practitioners, and dentists--adjusting for health status and other confounding variables. We quantified inequalities using the relative index of inequality (RII) using Poisson regression. The contribution of concession cards and PHI in promoting equity/inequity was examined using mediating models. RESULTS: There was equality in the use of GP services, but socio-economically advantaged women were more likely than disadvantaged women to use specialist (RII=1.41, 95% CI:1.26-1.58), allied health (RII=1.21,1.12-1.30), alternative health (RII=1.29,1.13-1.47) and dental services (RII=1.61,1.48-1.75) after adjusting for need, and they were less likely to visit hospital doctors (RII=0.74,0.57-0.96). Concession cards reduced socio-economic inequality in GP but not specialist care. Inequality in dental and allied health services was partly explained by inequalities in PHI. CONCLUSIONS AND IMPLICATIONS: Substantial socio-economic inequity exists in use of specialist and non-medical ambulatory care in Australia. This is likely to exacerbate existing health inequalities, but is potentially amenable to change. PMID- 19811485 TI - Matching health needs of refugee children with services: how big is the gap? AB - OBJECTIVES: To document the health needs of refugee children accessing comprehensive refugee health services in New South Wales (NSW), to match needs with available services and establish gaps in services. METHODS: We collated clinical data on all children aged under 14 years attending the three refugee specific clinics seeing children in NSW in 2005. We compared these data to the number of refugee children settling in NSW in 2005. RESULTS: NSW received 1,557 refugee children (<14 years) in 2005. Around one in five (n=331) was seen in a refugee specific clinic. Most were asymptomatic. Of those tested, 25% had anaemia, 27% were serology positive for schistosomiasis, 16% had evidence of current or recent malaria, 25% were tuberculin skin test positive, 69% were hepatitis B non-immune and 20% had low vitamin D levels. Most children needed catch up immunisation. Other problems included chronic health, developmental and behavioural problems. Screening tests varied across sites. Follow up was problematic for most. CONCLUSIONS: A small proportion of refugee children arriving in NSW have access to comprehensive screening and assessment, in spite of significant health needs. There is variation in screening practices, and follow up is poor. There is a high pick up rate for diseases of personal and public health significance. IMPLICATIONS: There is a strong moral and public health imperative to provide appropriately resourced, culturally competent and comprehensive health care to optimise refugee children's wellbeing. PMID- 19811486 TI - Fruit consumption among people living in a high deprivation New Zealand neighbourhood. AB - OBJECTIVE: To investigate fruit consumption in a high deprivation population in New Zealand. METHOD: In 2007, 99 door-to-door interviews were conducted in a high deprivation neighbourhood in Auckland with a focus on measuring self-reported fruit consumption. RESULTS: On average, participants reported eating a serving of fresh fruit five to six times per week. At the time of the interview, more than a third of participants (38%) did not have any fresh fruit in the house and 60% of respondents reported that in the past month they had thrown out fruit between one to four times per week because it was considered to be past its best in terms of eating quality. Fruit juice was consumed on average one to two times per week. Self-efficacy for fruit consumption was positively associated with consumption. Relative to participants with lower levels of self-efficacy for fruit consumption, those with higher levels of self-efficacy were more likely to achieve the target of consuming two or more servings of fruit daily. CONCLUSION: Strategies that aim to increase self-efficacy beliefs for fruit consumption may contribute to improving compliance with the recommended two or more servings daily. Together with strategies that give consideration to the social and cultural context and community level interventions (involving schools, churches and local community groups) they represent a holistic approach that is likely to be necessary for improving fruit consumption in high deprivation populations. PMID- 19811487 TI - Is lack of retail competition in the grocery sector a public health issue? AB - OBJECTIVES: The economic implications of a lack of competition in the grocery retail sector are hotly contested. However, there are also significant health implications of such anti-competitive practices that seldom receive attention. This paper hopes to draw attention to the potential public health issues that arise as a result of lack of competition in the grocery retail sector. METHOD: Relevant supporting literature was reviewed to explore the possible effects of market concentration on various health outcomes. RESULTS: High retailer concentration may adversely affect affordability, accessibility, quality, and choice of healthy food options to consumers. In turn this has significant implications for public health. IMPLICATIONS: Unless these upstream factors are addressed through the development of healthy competition, policy public health programs aimed purely at encouraging the public to consume higher quantities of healthful foods may be rendered ineffective. PMID- 19811488 TI - Surveillance of STI risk behaviour among young people attending a music festival in Australia, 2005-08. AB - OBJECTIVE: To explain rising rates of sexually transmitted infections it is necessary to monitor trends among high risk groups, such as youth. Surveillance of risk behaviours and testing among a variety of populations in different settings is required. We monitored self-reported sexual behaviour among music festival attendees. METHODS: Cross-sectional studies of young people's behaviour were conducted annually at a music festival between 2005 and 2008 using self administered questionnaires. Logistic regression, adjusted for age and gender, determined trends in risk behaviours. RESULTS: More than 5,000 questionnaires were completed. The proportion reporting multiple sexual partners in the past year remained stable from 2005 to 2008 and condom use with these partners increased. Reporting a new sexual partner in the past three months decreased, while condom use with new partners increased. Reporting a casual sexual partner increased and condom use with casual partners remained stable. Reporting a recent STI test increased from 23% in 2006 to 32% in 2008. CONCLUSIONS AND IMPLICATIONS: Despite increases in STI notifications, most risk behaviours are decreasing in this group, possibly as a function of increased STI testing. Music festivals are a useful setting for monitoring behaviour trends within a sub-population of young people at relatively high risk of STIs. PMID- 19811489 TI - Insect repellents and sunscreen: implications for personal protection strategies against mosquito-borne disease. AB - OBJECTIVE: To determine the protection times provided by insect repellent and sunscreen in combined formulations against biting mosquitoes. To determine if concurrent use of repellent and sunscreen influenced protection times. METHODS: Insect repellent containing comparable concentrations of N, N-diethyl-3 methylbenzamide (DEET) with and without sunscreen were tested on human skin to determine the mean protection time (MPT) against Aedes aegypti (L.) in the laboratory. Further trials were undertaken to determine the effect on MPT of sunscreen reapplication over repellent every two hours. RESULTS: There was no significant difference in the MPT provided by 80% DEET with (MPT+/-SE=770+/-54.8 minutes) and without (MPT+/-SE=830+/-20.2 minutes) sunscreen or 7.14% DEET with (MPT+/-SE =240+/-15.5 minutes) and 6.98% DEET without (MPT+/-SE =230+/-18.4 minutes) sunscreen. Reapplication of sunscreen resulted in a significantly lower MPT of a 17.0% DEET formulation when sunscreen was reapplied concurrently (MPT+/ SE=330+/-25.2 minutes), compared with DEET alone (MPT+/-SE =400+/-12.7 minutes). CONCLUSIONS: When combined in a single formulation with sunscreen, the MPT provided by both high and low concentrations of DEET is not reduced. However, if sunscreen is reapplied over insect repellent, protection times can be reduced significantly. IMPLICATIONS: In areas of endemic mosquito-borne disease, the reapplication of a low concentration repellent and sunscreen formulation may provide the most effective protection from biting mosquitoes while minimising the risk of overexposure to DEET. PMID- 19811490 TI - Changes in solarium numbers in Australia following negative media and legislation. AB - OBJECTIVE: To monitor changes in the number of indoor tanning facilities in Australia's capital cities following widespread negative publicity and the introduction of legislation in some states. METHODS: An audit of listings under Solarium/Tanning Centres in the most recent hard copy Yellow Pages for Australia's capital cities was conducted, and results were compared with those from a previous audit from 2006. RESULTS: There was a 32% drop in solarium listings for Australia's capital cities between 2006 and 2008/09. In most cases, larger decreases were observed in states where legislation was introduced. CONCLUSIONS: Despite underestimating solarium numbers, regular audits of business listings can be a useful way of monitoring trends, particularly when more accurate figures are not available. Solarium numbers can decrease following a combination of negative publicity and legislation. IMPLICATIONS: To achieve and maintain predicted reductions in skin cancer incidence, mortality and costs to the health system, solarium legislation should be a priority for those states where it has not yet been introduced, accompanied by compliance monitoring and enforcement to ensure it is effective. PMID- 19811492 TI - The substance use profiles of a sample of obligatory and non-obligatory exercisers. PMID- 19811497 TI - Stratum corneum integrity as a predictor for peristomal skin problems in ostomates. AB - BACKGROUND: Peristomal skin problems are common, most often the result is disruption of the skin barrier and this may account for more than one in three visits to ostomy nurses. Therefore a specific assessment of individual risk factors relating to the skin barrier function would be of great interest. METHODS: Skin barrier integrity in ostomy patients with peristomal skin problems (PSP) was compared with that of ostomy patients with normal skin (controls) using transepidermal water loss (TEWL). Mechanical barrier disruption was determined by a tape stripping test and chemical barrier disruption [sodium lauryl sulphate (SLS) 0.25%]. RESULTS: Patients and controls had a highly significant increase in TEWL value in the peristomal area compared with nonperistomal contralateral abdominal skin (P < 0.0001 for both groups). The skin barrier of normal-looking contralateral skin of ostomates was found to be borderline impaired in patients with PSP compared with those without. A linear association was seen between the number of tape strips removed and TEWL for both cases and controls. Tape stripping suggested that patients with PSP had less resilient skin (P = 0.002). A significant difference in TEWL value between cases and controls was also seen for the SLS patch test on the dorsal skin (P = 0.02). CONCLUSION: Successive tape stripping, a situation analogous to the normal use of a pouching system, caused a higher degree of barrier damage more rapidly in patients with PSP, indicating an impaired mechanical quality of the barrier. The SLS exposure test suggested a generally increased susceptibility to irritant dermatitis as assessed by TEWL. Our findings suggest tape stripping and SLS testing may have a role as predictive tests to identify patients at risk of PSP. PMID- 19811498 TI - Significance of thymidylate synthase for resistance to pemetrexed in lung cancer. AB - Pemetrexed (MTA) is a multitargeted antifolate with promising clinical activity in lung cancer. We exposed the small cell lung cancer cell line PC6 to stepwise increasing pemetrexed concentrations of 0.4, 1.6, and 4.0 microm, and established three pemetrexed-resistant lung cancer cell lines: PC6/MTA-0.4, PC6/MTA-1.6, and PC6/MTA-4.0 cells. To investigate the mechanisms of acquired resistance to pemetrexed, we measured the expression levels of the thymidylate synthase (TS), reduced folate carrier (RFC), and folylpoly-gamma-glutamate synthetase (FPGS) genes. TS gene expression was significantly increased in PC6/MTA-1.6 and PC6/MTA 4.0 cells relative to parental cells in a pemetrexed dose-dependent manner. In contrast, the levels of RFC gene expression in PC6/MTA-0.4 cells and FPGS in PC6/MTA-1.6 cells were significantly decreased, whereas the levels of both genes were restored in PC6/MTA-4.0 cells. Knockdown of TS expression using siRNA enhanced pemetrexed cytotoxicity in PC6/MTA-4.0 cells. The expression level of the TS gene was significantly correlated with the concentration of pemetrexed for 50% cell survival (IC(50)) in 11 non-small cell lung cancer cell lines. These results suggest that the alteration of molecular pharmacological factors in relation with pemetrexed resistance is dose-dependent, and that up-regulation of the expression of the TS gene may have an important role in the acquired resistance to pemetrexed. In addition, TS may be a predictive marker for pemetrexed sensitivity in lung cancer. PMID- 19811499 TI - Independent and cooperative roles of tumor necrosis factor-alpha, nuclear factor kappaB, and bone morphogenetic protein-2 in regulation of metastasis and osteomimicry of prostate cancer cells and differentiation and mineralization of MC3T3-E1 osteoblast-like cells. AB - The molecular mechanisms involved in prostate cancer (PC) metastasis and bone remodeling are poorly understood. We recently reported that phosphoinositide 3 kinase (PI3K)/protein kinase B (Akt) mediates transcriptional regulation and activation of bone morphogenetic protein (BMP)-2 signaling by nuclear factor (NF) kappaB in bone metastatic prostate cancer cells. In the present study, we demonstrate that NF-kappaB, whether activated by recombinant human tumor necrosis factor (TNF)-alpha or by ectopic expression of the p65 subunit, is involved in extracellular matrix adhesion and invasion of osteotropic PC-3 and C4-2B, but not LNCaP, cells. The enhanced metastatic potential was associated with transcriptional upregulation of osteopontin, osteocalcin, and collagen IA1 in osteotropic PC cells, suggesting their role in osteomimicry of PC cells. Unlike BMP-4, BMP-2 protein enhanced the invasive properties of C4-2B cells, but not in LNCaP cells. Also, this effect was nullified by Noggin. In addition, BMP-2 mediates TNF-alpha-induced invasion of C4-2B cells in a NF-kappaB-dependent fashion. TNF-alpha or conditioned media (CM) of TNF-alpha-stimulated C4-2B cells upregulated BMP-2 and BMP-dependent Smad transcripts and inhibited receptor activator of NF-kappaB ligand transcripts in RAW 264.7 preosteoclast cells, respectively, implying that this factor may contribute to suppression of osteoclastogenesis via direct and paracrine mechanisms. In contrast, CM of TNF alpha-stimulate or BMP2-stimulated C4-2B cells induced in vitro mineralization of MC3T3-E1 osteoblast cells in a BMP-2-dependent and NF-kappaB-dependent manner, respectively. Taken together, the results suggest that mutual interactions between these factors may be pivotal not only in enhancing the osteomimicry and metastatic potential of PC cells, but also in bone remodeling and in shifting the balance from osteoclastogenesis towards osteoblastogenesis. PMID- 19811500 TI - Autophagy supports Candida glabrata survival during phagocytosis. AB - The opportunistic human fungal pathogen Candida glabrata is confronted with phagocytic cells of the host defence system. Survival of internalized cells is thought to contribute to successful dissemination. We investigated the reaction of engulfed C. glabrata cells using fluorescent protein fusions of the transcription factors CgYap1 and CgMig1 and catalase CgCta1. The expression level and peroxisomal localization of catalase was used to monitor the metabolic and stress status of internalized C. glabrata cells. These reporters revealed that the phagocytosed C. glabrata cells were exposed to transient oxidative stress and starved for carbon source. Cells trapped within macrophages increased their peroxisome numbers indicating a metabolic switch. Prolonged phagocytosis caused a pexophagy-mediated decline in peroxisome numbers. Autophagy, and in particular pexophagy, contributed to survival of C. glabrata during engulfment. Mutants lacking CgATG11 or CgATG17, genes required for pexophagy and non-selective autophagy, respectively, displayed reduced survival rates. Furthermore, both CgAtg11 and CgAtg17 contribute to survival, since the double mutant was highly sensitive to engulfment. Inhibition of peroxisome formation by deletion of CgPEX3 partially restored viability of CgATG11 deletion mutants during engulfment. This suggests that peroxisome formation and maintenance might sequester resources required for optimal survival. Mobilization of intracellular resources via autophagy is an important virulence factor that supports the viability of C. glabrata in the phagosomal compartment of infected innate immune cells. PMID- 19811502 TI - Variable expression of surface-exposed polymorphic membrane proteins in in vitro grown Chlamydia trachomatis. AB - The hypothesized variable expression of polymorphic membrane proteins (PmpA-PmpI) in Chlamydia trachomatis-infected patients was tested by examination of the expression of each Pmp subtype in in vitro-grown C. trachomatis. A panel of monospecific polyclonal and monoclonal antibodies was used to demonstrate surface exposure of Pmps of each subtype by differential immunofluorescence (IF) with and without prior detergent permeabilization of paraformaldehyde-fixed inclusions and for selected Pmps by immunogold labelling. Although specific transcript was detected for each pmp gene late in development, IF experiments with Pmp subtype specific antibodies reveal that a number of inclusions in a single infection do not express Pmps of a given subtype. Coexpression experiments suggest that pmp genes are shut off independently from one another in non-expressing inclusions, i.e. different inclusions are switched off for different Pmps. Overall, these studies establish the existence of an efficient shutoff mechanism independently affecting the expression of each member of the pmp gene family in in vitro-grown C. trachomatis. Like other paralogous gene families of bacterial pathogens, the pmp gene family of C. trachomatis may serve the critical dual function of a highly adaptable virulence factor also providing antigenic diversity in the face of the host adaptive immune response. PMID- 19811501 TI - ATP-dependent activation of an inflammasome in primary gingival epithelial cells infected by Porphyromonas gingivalis. AB - Production of IL-1beta typically requires two-separate signals. The first signal, from a pathogen-associated molecular pattern, promotes intracellular production of immature cytokine. The second signal, derived from a danger signal such as extracellular ATP, results in assembly of an inflammasome, activation of caspase 1 and secretion of mature cytokine. The inflammasome component, Nalp3, plays a non-redundant role in caspase-1 activation in response to ATP binding to P2X(7) in macrophages. Gingival epithelial cells (GECs) are an important component of the innate-immune response to periodontal bacteria. We had shown that GECs express a functional P2X(7) receptor, but the ability of GECs to secrete IL-1beta during infection remained unknown. We find that GECs express a functional Nalp3 inflammasome. Treatment of GECs with LPS or infection with the periodontal pathogen, Porphyromonas gingivalis, induced expression of the il-1beta gene and intracellular accumulation of IL-1beta protein. However, IL-1beta was not secreted unless LPS-treated or infected cells were subsequently stimulated with ATP. Conversely, caspase-1 is activated in GECs following ATP treatment but not P. gingivalis infection. Furthermore, depletion of Nalp3 by siRNA abrogated the ability of ATP to induce IL-1beta secretion in infected cells. The Nalp3 inflammasome is therefore likely to be an important mediator of the inflammatory response in gingival epithelium. PMID- 19811503 TI - Short-term frovatriptan for the prevention of difficult-to-treat menstrual migraine attacks. AB - The efficacy of a 6-day regimen of frovatriptan for menstrual migraine (MM; attacks starting on day -2 to +3 of menses) prevention in women with difficult-to treat MM was assessed. Women with a documented inadequate response to triptans for acute MM treatment were included in this placebo-controlled, parallel-group trial. Women were randomized to double-blind treatment for three perimenstrual periods (PMPs) with either frovatriptan 2.5 mg (q.d. or b.i.d.) or placebo initiated 2 days before anticipated MM. The efficacy analysis included 410 women with 85% completing three double-blind PMPs. The mean number of headache-free PMPs was 0.92 with frovatriptan b.i.d., 0.69 with frovatriptan q.d. and 0.42 with placebo [P < 0.001 (b.i.d.) and P < 0.02 (q.d.) vs. placebo]. When migraine occurred, severity was reduced with frovatriptan q.d. (P < 0.001) and b.i.d. (P < 0.001) vs. placebo. Both frovatriptan regimens were well tolerated. In women with difficult-to-treat MM, a 6-day regimen of frovatriptan significantly reduced MM incidence and severity. PMID- 19811504 TI - The prevalence of premonitory symptoms in paediatric migraine: a questionnaire study in 103 children and adolescents. AB - The prevalence and characterization of premonitory symptoms have not been rigorously studied in children and adolescents. Using a questionnaire, we retrospectively studied the prevalence of 15 predefined premonitory symptoms in a clinic-based population. In 103 children and adolescents fulfilling the International Classification of Headache Disorders, 2nd edn criteria for paediatric migraine, at least one premonitory symptom was reported by 69 (67%). The most frequently reported premonitory symptoms were face changes, fatigue and irritability. The mean number of premonitory symptoms reported per subject was 1.8 (median 2.2). Age, migraine subtype (with or without aura) and mean attack frequency per month had no effect on the mean number of premonitory symptoms reported per subject. In conclusion, premonitory symptoms are frequently reported by children and adolescents with migraine. Face changes seem to be a premonitory symptom peculiar to paediatric migraine. PMID- 19811505 TI - Sphenoid fungus balls: clinical presentation and long-term follow-up in 24 patients. AB - Fungus balls are a non-invasive form of fungal infection involving the maxillary sinus in most cases. Sphenoid sinus fungus balls (SSFB) are rare and their clinical presentation is not well described. We intended to define the clinical presentation of sphenoid fungus balls, and retrospectively reviewed 24 cases of SSFB seen at our institution over a 10-year period, identified through pathological reports. Presenting symptoms were separated into three groups: headache, rhinological and asymptomatic. Headaches were subdivided into acute and chronic, unilateral and diffuse. Radiological clues leading to diagnosis were reviewed. Prognosis was determined from medical files or by phone calls. Sixty seven per cent of patients were female. The mean age at presentation was 65 years. Sixty-two per cent presented with headache, 36% unilateral, mainly in the first trigeminal branch territory. Rhinological symptoms were seen in 21%. In 16% of patients the SSFB was asymptomatic and found during routine tests. SSFB, even if non-invasive, did lead to recurrent bacterial infections and central nervous system complications in three patients. Of 15 patients presenting with headache, 10 were significantly improved post surgery. The prognosis is good, with no recurrence of fungal infection after a main follow-up of 2.3 years. Our study underlines that SSFB present with headaches, often unilateral and in the fronto orbital region. Proper imaging of the sphenoid sinus is useful in patients with unexplained headache. The neurologist has to be aware of radiological clues suggesting fungal sinus infection, since surgery is the main treatment, with good prognosis and frequent resolution of headaches. PMID- 19811506 TI - Atomic interactions and profile of small molecules disrupting protein-protein interfaces: the TIMBAL database. AB - Growing evidence of the possibility of modulating protein-protein interactions with small molecules is opening the door to new approaches and concepts in drug discovery. In this paper, we describe the creation of TIMBAL, a hand-curated database holding an up to date collection of small molecules inhibiting multi protein complexes. This database has been analysed and profiled in terms of molecular properties. Protein-protein modulators tend to be large lipophilic molecules with few hydrogen bond features. An analysis of TIMBAL's intersection with other structural databases, including CREDO (protein-small molecule from the PDB) and PICCOLO (protein-protein from the PDB) reveals that TIMBAL molecules tend to form mainly hydrophobic interactions with only a few hydrogen bonding contacts. With respect to potency, TIMBAL molecules are slightly less efficient than an average medicinal chemistry hit or lead. The database provides a resource that will allow further insights into the types of molecules favoured by protein interfaces and provide a background to continuing work in this area. Access at http://www-cryst.bioc.cam.ac.uk/timbal. PMID- 19811507 TI - Interaction energy analysis of peptide can predict the possibilities of mimetics by its retroinverso isomer. AB - It has been previously reported that the retroinverso analog of S peptide cannot mimic the S peptide, whereas the retroinverso analog of foot-and-mouth disease virus antigen can mimic the foot-and-mouth disease virus antigen. The structures of S peptide, foot-and-mouth disease virus antigen, and their retroinverso analogs are known. Here, we have attempted to explain the structural basis of mimetics at the level of atomic interactions by elaborating upon the Guptasarma's hypothesis. Using interaction energy analysis of S peptide and foot-and-mouth disease virus antigen, we propose that if the energy of the CO and NH backbone atoms' non-covalent interactions with all other atoms is negligible as compared with the energy of other non-covalent interactions, then the retroinverso isomer can mimic the original peptide/protein. Previous work has established that the structure of the inverso analog of a protein will be the mirror image of the protein, and it will only recognize the respective mirror image substrate/binding partner. The retro peptide conformation that can be superimposed on all side chains in any conformation of the original peptide does not exist in the conformational space of the peptides. PMID- 19811508 TI - The concordance between serum anti-Mullerian hormone and testosterone concentrations depends on duration of hCG stimulation in boys undergoing investigation of gonadal function. AB - BACKGROUND: In boys undergoing investigation of gonadal function, the relationship between a single measurement of serum anti-Mullerian hormone (AMH) and hCG stimulated serum testosterone is unclear. AIM: The aim of the study was to assess concordance between serum AMH and testosterone concentrations following hCG stimulation of two different durations. METHODS: Samples from 284 children (M : F, 154 : 130) with a median age of 8 years (10th, 90th centiles, 0.25, 14) were used to establish an AMH reference range. Clinical data were reviewed in boys undergoing investigation of gonadal function and who had an AMH measurement and a hCG stimulated (3-day or 3-week) (n = 26) testosterone. Of these 26 boys, 11 had combined genital anomalies, whereas the rest had conditions such as isolated hypospadias, undescended testes or microphallus. Normal testosterone response to hCG stimulation was defined as a level greater than 3.5 nmol at day 4 and 9.5 nmol/l at day 22. RESULTS: In the reference group, the 5th centile AMH for boys below 1 year was 215 pmol/l and between 1 and 8 years 180 pmol/l. The 95th centile for girls for these respective age groups was 30 pmol/l and 25 pmol/l. In those cases where serum testosterone concentrations were available at day 1, day 4 and day 22 of the 3 week-hCG test, five cases had a normal serum testosterone at day 4 and three cases only showed such a response by day 22. In those where serum AMH was less than 180 pmol/l, a poor testosterone response of less than 3.5 nmol was observed in approximately seven of eight (88%) cases with a 3-day hCG stimulation test or the 3-week test. An AMH of greater than 180 pmol/l was associated with a normal testosterone response at day 4 in 10 out of 15 (67%) cases and at day 22 in eight of 11 (73%) cases. However, a low serum testosterone concentration of less than 3.5 nmol after the 3-day hCG test was only associated with a likelihood of a low AMH in three of eight (37%) cases. With the 3-week hCG test, a low day 22 testosterone of 9.5 mmol/l or less was associated with a low AMH of 180 pmol/l or less in four of seven (57%) cases. CONCLUSION: In boys undergoing investigation of gonadal function, the concordance between AMH and testosterone is better at day 22 than day 4. A normal AMH may provide useful information on overall testicular function but does not exclude the need for an hCG stimulation test. PMID- 19811509 TI - 3 Tesla magnetic resonance imaging with and without corticotropin releasing hormone stimulation for the detection of microadenomas in Cushing's syndrome. AB - OBJECTIVE: We sought to determine if higher resolution 3 Tesla (T) magnetic resonance imaging (MRI) with or without ovine corticotropin releasing hormone (o CRH) stimulation would increase the sensitivity for detection of pituitary microadenomas in ACTH-dependent Cushing's syndrome (CS). DESIGN AND PATIENTS: We prospectively identified 23 patients over a 2-year period with clinical and biochemical evidence of ACTH-dependent CS with no lesion (n = 11) or equivocal lesion (n = 10) on 1.5T MRI. Subsequently, two additional MRIs were performed in random order: 3T nonstimulated MRI or 3T MRI with o-CRH in all patients. Three neuroradiologists reviewed all examinations in a randomized blinded fashion. Patients were divided into four groups, depending on the outcome of their evaluation and treatment for CS. Two patients had to be excluded, and so we report on 21 subjects. MEASUREMENTS AND RESULTS: Both 3T MRI without (P < 0.016) and with o-CRH stimulation (P < 0.013) was significantly more sensitive for detection of pituitary microadenomas than 1.5T MRI for Group 1 (definitive proof of Cushing's disease, n = 10). Group 2 (those in group 1, plus three patients where dynamic/invasive testing suggested pituitary source) also showed a significant (P < 0.012) advantage for 3T. There was no difference between the 3T and the 3T o-CRH examinations for any of the pulse sequences. We did not observe a statistically significant difference in other patient groups [patients with recurrent CD (n = 6) and patients with ectopic CS (n = 2)]. CONCLUSIONS: The results of our prospective blinded studies suggest that 3T MRI of pituitary gland should be considered in evaluation of patients with ACTH-dependent CD when 1.5T imaging is negative or equivocal. PMID- 19811510 TI - Social and behavioral risk factors for maxillary incisor trauma in an adolescent Arab population. AB - BACKGROUND/AIM: The majority of dental traumas are due to falls or blows during regular activity, personal interaction and play inside or outside the home, suggesting that behaviors associated with certain social conditions are risk factors. Our purpose was to explore that hypothesis. MATERIALS AND METHODS: We examined a population-based sample of 1583 13-14-year-old subjects in a classroom setting. Incisor trauma was scored according to the National Institute of Dental Research index, and overjet was measured to the nearest 0.5 mm. Presence of mesial migration and/or loss of first molars and/or open caries in permanent teeth were recorded and categorized as increased caries experience. Body mass index (BMI) was calculated from standardized measurements of height and weight, as weight in kg height(-1) in m(2). Obesity was scored if BMI exceeded 30. Information on participation in physical activities, family income, and number of siblings was collected through subject and family interview. RESULTS AND CONCLUSIONS: Univariate logistic regression detected reduced trauma risk among subjects of high income families (P = 0.009) and among subjects attending private schools (P = 0.026). Sports and physical activities were more prevalent in boys than in girls (P = 0.000) and associated with trauma in the sample as a whole (P = 0.001), but not separately in boys and girls. No effect was detected of obesity, number of siblings and increased caries experience. Univariate analyzes also detected increased trauma risk in boys (P = 0.000) and in subjects with increasing intervals of overjet (P = 0.000). Following use of multiple regressions with forward selection, only male gender, increasing overjet, and high family income were included in the final prediction model. Our findings suggest that behaviors associated with high family income and private school attendance reduce the risk of incisor trauma among adolescents in Kuwait, and that the gender difference in sports activities is an unlikely explanation for the gender difference in trauma. PMID- 19811511 TI - Contradictions in the treatment of traumatic dental injuries and ways to proceed in dental trauma research. AB - Almost all treatment procedures used for dental traumas are still today not evidence-based, a fact, which makes it difficult to analyse the long-term outcome of healing and its relationship to treatment. Crown fractures with extensive dentin exposure represent a dominant injury in the permanent dentition. Accepted treatment philosophy is dentin coverage (dental liner and/or dentin bonded restoration) to prevent bacteria penetration into the pulp. Today there is, apart from deep proximal fractures, no evidence that this treatment is necessary to protect the pulp. In case of luxation injuries, the accepted treatment principles appear to be anatomically correct repositioning, stabilization with a splint and sometimes antibiotic coverage. In clinical studies, these principles could not be proven to optimize either periodontal or pulpal healing, the explanation possibly being that both reposition and application of splints in certain cases add extra damage to the pulp and periodontal ligament. In case of root fractures with dislocation, fast and optimal repositioning and rigid long-term splinting (i.e. 3 months) have been considered the principle of treatment. However, a recent clinical study has shown that short-term splinting with a semi-rigid splint appears to optimize fracture healing. In tooth avulsion with subsequent replantation, cleansing of the root surface for contamination and systemic antibiotics has been considered essential for pulp and periodontal healing. These treatment concepts have been derived from experimental studies in animals. However, their importance could not be verified in large clinical studies. Ideally, randomized clinical studies are needed in the future for selected trauma types. The influences of repositioning, splinting and the role of infection and antibiotics should be further investigated. However, for ethical reasons, it will be difficult to perform randomized studies on trauma victims and we will be forced in the future to rely on experimental animal studies supported by clinical observational studies. PMID- 19811512 TI - Recurrent acute necrotizing encephalopathy following influenza A in a genetically predisposed family. AB - Acute necrotizing encephalopathy (ANE) typically affects young, healthy children who develop rapid-onset severe encephalopathy triggered by viral infections. This disease is more commonly reported in Japan but occurs worldwide, although it remains under-recognized in Western countries. An autosomal dominant form, ANE1, was recently identified. We report the details of a 9-year-old Caucasian female who experienced recurrent ANE episodes at the ages of 9 months and 9 years. Brain magnetic resonance imaging findings were characteristic of ANE during both episodes, although more extensive in the recent episode, which resulted in severe neurological sequelae; influenza A was identified on bronchoalveolar lavage during this episode. Interestingly, there was evidence of peripheral polyneuropathy during the recent episode, which has not previously been described in sporadic ANE. Both the patient and her mother, who had also had postviral polyneuritis in the past, harbour a mutation in Ran-binding protein 2 (RANBP2); this occurred de novo in the mother and confers genetic susceptibility to ANE. Our case suggests that recurrent disease and/or an expanded clinical phenotype raises the possibility of ANE1; positive family history, although supportive, is not necessary as the mutation can occur de novo. Increased awareness may lead to earlier recognition and better treatment options. PMID- 19811513 TI - Upper limb activity measures for 5- to 16-year-old children with congenital hemiplegia: a systematic review. AB - AIM: This systematic review aimed to compare the validity, reliability, evaluative validity, and clinical utility of upper limb activity measures for children aged 5 to 16 years with congenital hemiplegia. METHOD: Electronic databases were searched to identify assessments that measure upper limb activity available for use and for which published validity and reliability data for the population are obtainable. Assessment items were coded according to the International Classification of Functioning, Disability and Health (ICF) categories to determine if at least 35% of the assessment items fell within the activity component of the ICF. Assessments that met these criteria were included in the review. RESULTS: Thirty-eight measures were identified, and five met the inclusion criteria. The best measure of unimanual capacity was the Melbourne Assessment of Unilateral Upper Limb Function (MUUL); however, the Shriners Hospital Upper Extremity Evaluation (SHUEE) and the Quality of Upper Extremity Skills Test (QUEST) could also be considered, depending on the type of information required. The performance-based measure of bimanual upper limb activity in children with hemiplegia with the best psychometric properties was the Assisting Hand Assessment (AHA). The ABILHAND-Kids is a parent-report, performance-based questionnaire with excellent clinical utility and psychometric properties. INTERPRETATION: Clinicians may choose to use more than one of these measures to detect changes in unimanual or bimanual upper limb activity. PMID- 19811514 TI - Sporadic hemiplegic migraine and delayed cerebral oedema after minor head trauma: a novel de novo CACNA1A gene mutation. PMID- 19811515 TI - Relationship between neuromuscular body functions and upper extremity activity in children with cerebral palsy. AB - AIM: Our aim was to investigate the relationship between the dimensions of neuromuscular body function and elbow, forearm, and hand activity in the upper extremities in children/adolescents with spastic cerebral palsy (CP), within the framework of the World Health Organization International Classification of Functioning, Disability and Health. METHOD: Twenty-three participants (10 males, 13 females, mean age 13y, SD 3y, range 8-18y) with spastic CP (21 with hemiplegia, two with diplegia) at Manual Ability Classification System levels I to III participated in the study. Neuromuscular body function measures were (1) muscle strength in the elbow, forearm, and grip, (2) muscle tone in elbow flexors and forearm supinators, (3) active supination range and elbow extension range, and (4) force control at submaximal level in elbow flexion. Activity measures were actual use of the affected hand in bimanual activities (Assisting Hand Assessment) and instructed use of the affected hand (Melbourne Assessment of Unilateral Upper Limb Function). RESULTS: Nearly all the neuromuscular body function variables were significantly correlated with activity. The combination of active supination range and strength explained 74% of the variance in actual use, and the combination of active supination range and force control explained 74% of the variance in instructed use. INTERPRETATION: In high-functioning children and adolescents with CP, limited active supination range and difficulties in generating and modulating force are strongly related to limitations in hand activity. Further studies are needed to establish cause and effect in this relationship. PMID- 19811516 TI - Development and validation of item sets to improve efficiency of administration of the 66-item Gross Motor Function Measure in children with cerebral palsy. AB - AIM: To develop an algorithmic approach to identify item sets of the 66-item version of the Gross Motor Function Measure (GMFM-66) to be administered to individual children, and to examine the validity of the algorithm for obtaining a GMFM-66 score. METHOD: An algorithmic approach was used to identify item sets of the GMFM-66 (GMFM-66-IS) using data from 95 males and 79 females with cerebral palsy (CP; mean age 14y 7mo, SD 1y 8mo, range 12y 7mo to 17y 8mo). The GMFM-66-IS scores were then validated using combined data from three Dutch studies involving 134 males and 92 females with CP (mean age 7y, SD 4y 6mo, range 1y 4mo to 13y 8mo), representing all levels of the Gross Motor Function Classification System. RESULTS: The final algorithm contains three decision items from the GMFM-66 that determine which one of four item sets to administer. The GMFM-66-IS has excellent agreement with the full GMFM-66 both at a single assessment (intraclass correlation coefficient [ICC]=0.994, 95% confidence intervals [CI] 0.993-0.996) and across repeat assessments (ICC=0.92, 95% CI 0.89-0.95). INTERPRETATION: The GMFM-66-IS is a promising alternative to the full GMFM-66. Users should be consistent in their choice of measure (GMFM-66 or GMFM-66-IS) on repeat testing and clearly identify which method was used. PMID- 19811517 TI - Development of the Hypertonia Assessment Tool (HAT). PMID- 19811518 TI - Assessment and correction of skinfold thickness equations in estimating body fat in children with cerebral palsy. AB - AIM: To assess the accuracy of skinfold equations in estimating percentage body fat in children with cerebral palsy (CP), compared with assessment of body fat from dual energy X-ray absorptiometry (DXA). METHOD: Data were collected from 71 participants (30 females, 41 males) with CP (Gross Motor Function Classification System [GMFCS] levels I-V) between the ages of 8 and 18 years. Estimated percentage body fat was computed using established (Slaughter) equations based on the triceps and subscapular skinfolds. A linear model was fitted to assess the use of a simple correction to these equations for children with CP. RESULTS: Slaughter's equations consistently underestimated percentage body fat (mean difference compared with DXA percentage body fat -9.6/100 [SD 6.2]; 95% confidence interval [CI] -11.0 to -8.1). New equations were developed in which a correction factor was added to the existing equations based on sex, race, GMFCS level, size, and pubertal status. These corrected equations for children with CP agree better with DXA (mean difference 0.2/100 [SD=4.8]; 95% CI -1.0 to 1.3) than existing equations. INTERPRETATION: A simple correction factor to commonly used equations substantially improves the ability to estimate percentage body fat from two skinfold measures in children with CP. PMID- 19811519 TI - Survival of individuals with cerebral palsy receiving continuous intrathecal baclofen treatment: a matched-cohort study. AB - AIM: To determine whether intrathecal baclofen (ITB) changes mortality risk in persons with cerebral palsy (CP). METHOD: Records were reviewed for all persons with CP who were managed with ITB for hypertonicity at a specialty hospital in Minnesota between May 1993 and August 2007. A comparison cohort was randomly selected from clients of the California Department of Developmental Services who were initially evaluated between 1987 and 1990 and were matched to those with ITB for age, sex, Gross Motor Function Classification System (GMFCS) level, presence or absence of epilepsy, and feeding-tube use. Survival probabilities were estimated using the Kaplan-Meier method, and differences were tested via log rank. RESULTS: Three hundred and fifty-nine persons with CP (202 males, 157 females) receiving ITB for hypertonicity (mean age 12y 8mo, SD 7y 9mo, range 3y 1mo to 39y 9mo) were matched to 349 persons without ITB pumps (195 males, 154 females; mean age 12y 7mo, SD 8y 4mo, range 2y 7mo to 40y). The proportion of patients at different GMFCS levels in the ITB and in the non-ITB cohorts, respectively, was as follows: level II 3% and 3%, level III 16% and 16%, level IV 38% and 37%, and level V 43% and 44%. Survival at 8 years of follow-up was 92% (SD 1.9%) in the ITB cohort and 82% (SD 2.4%) in the non-ITB cohort (p<0.001). After adjustment to account for recent trends in improved survival in CP, 8-year survival in the non-ITB cohort was 88%, which was not significantly different from the ITB cohort (p=0.073). INTERPRETATION: ITB therapy does not increase mortality in individuals with CP and may suggest an increase in life expectancy. PMID- 19811521 TI - Ultrasound imaging: a promising method for assessment of body composition. PMID- 19811520 TI - White matter abnormalities and dystonic motor disorder associated with mutations in the SLC16A2 gene. AB - AIM: Mutations in the SLC16A2 gene have been implicated in Allan-Herndon-Dudley syndrome (AHDS), an X-linked learning disability* syndrome associated with thyroid function test (TFT) abnormalities. Delayed myelination is a non-specific finding in individuals with learning disability whose genetic basis is often uncertain. The aim of this study was to describe neuroimaging findings and neurological features in males with SLC16A2 gene mutations. METHOD: We reviewed brain magnetic resonance imaging (MRI) findings and neurological features in a cohort of five males aged between 1 year 6 months and 6 years (median 4y) from four families harbouring SLC16A2 gene mutations. RESULTS: The participants presented aged between 4 and 9 months with initial hypotonia and subsequent spastic paraparesis with dystonic posturing and superimposed paroxysmal dyskinesias. Dystonic cerebral palsy was the most common initial clinical diagnosis, and AHDS was suspected only retrospectively, considering the characteristically abnormal thyroid function tests, with high serum tri iodothyronine (T(3)), as the most consistent finding. Brain MRI showed absent or markedly delayed myelination in all five participants, prompting the suspicion of Pelizaeus-Merzbacher disease in one patient. INTERPRETATION: Our findings indicate a consistent association between defective neuronal T(3) uptake and delayed myelination. SLC16A2 involvement should be considered in males with learning disability, an associated motor or movement disorder, and evidence of delayed myelination on brain MRI. Although dysmorphic features suggestive of AHDS are not always present, T(3) measurement is a reliable screening test. PMID- 19811522 TI - Reading performance correlates with white matter properties in preterm and term children. PMID- 19811523 TI - Social functioning and communication in children with cerebral palsy: association with disease characteristics and personal and environmental factors. AB - AIM: The objective of this longitudinal study was to describe the course of social functioning and communication in children with cerebral palsy (CP) over a 3-year period, its difference with the normative course, and its relationship with disease characteristics and personal and environmental factors. METHOD: Participants in this study were 110 children with CP (70 males, 40 females) with a mean age of 11 years and 3 months (SD 1y 8mo). Social functioning and communication were measured with the Vineland Adaptive Behavior Scales. Comparisons were made with normative data; data were analysed with generalized estimating equations. According to the Gross Motor Function Classification System (GMFCS), 50 of the 110 children were categorized as GMFCS level I, 16 as level II, 13 as level III, 13 as level IV, and 18 as level V. RESULTS: The course of social functioning over a 3-year period showed an increase in restrictions in children with CP (p<0.001). Restrictions in communication increased more in children with the most severe forms of CP (p<0.001). In addition to disease characteristics (GMFCS category, presence of epilepsy, and speech problems), personal factors (externalizing behaviour problems) and environmental factors (having no siblings, low parental level of education, and parental stress) were associated with greater restrictions in social functioning and communication. INTERPRETATION: The results indicate that it is important to focus not only on the medical treatment of children with CP, but also on their behavioural problems and social circumstances, and to support the parents so that social functioning and communication in these children may be improved. PMID- 19811524 TI - Two hands are better than one: bimanual skill development in children with hemiplegic cerebral palsy. PMID- 19811525 TI - Palliative care, public health and justice: setting priorities in resource poor countries. AB - Many countries have not considered palliative care a public health problem. With limited resources, disease-oriented therapies and prevention measures take priority. In this paper, I intend to describe the moral framework for considering palliative care as a public health priority in resource-poor countries. A distributive theory of justice for health care should consider integrative palliative care as morally required as it contributes to improving normal functioning and preserving opportunities for the individual. For patients requiring terminal care, we are guided less by principles of justice and more by the duty to relieve suffering and society's commitment to protecting the professional's obligation to uphold principles of beneficence, compassion and non abandonment. A fair deliberation process is necessary to allow these strong moral commitments to serve as reasons when setting priorities in resource poor countries. PMID- 19811526 TI - High plasma levels of the soluble receptor for advanced glycation endproducts in patients with symptomatic carotid atherosclerosis. AB - BACKGROUND: Advanced glycation endproducts (AGEs), particularly carboxymethyl(lysine)-adducts (CML), exert part of their cellular effects by binding to a receptor, named receptor for AGEs (RAGE). The soluble form of this receptor (sRAGE) has been shown to have an athero-protective role. We hypothesized the existence of a relationship between the AGE-RAGE axis and the occurrence of symptoms related to carotid atherosclerosis in nondiabetic conditions. MATERIALS AND METHODS: We evaluated plasma levels of CML and sRAGE (by ELISA), and tissue levels (tAGEs and tRAGE, semiquantitatively, by immunohistochemistry) in endarterectomy carotid plaque tissue in 29 nondiabetic patients. At the time of surgery, 10 patients were asymptomatic and 19 were symptomatic. RESULTS: Plasma levels of sRAGE were higher in symptomatic patients than in asymptomatic patients [median (interquartile range): 676 (394-858) pg mL( 1) vs. 347 (284-479) pg mL(-1), P = 0.009]. In symptomatic patients, plasma levels of sRAGE correlated positively with CML (r = 0.60, P < 0.01), C-reactive protein (CRP) (r = 0.618, P < 0.01) and fibrinogen (r = 0.522, P<0.005), while in asymptomatic patients, no correlation was observed. Although tissue and plasma levels of AGEs and RAGE did not correlate between each other, tAGEs and tRAGE were also positively correlated only in symptomatic patients (chi(2) = 8.93, P = 0.003). CONCLUSIONS: Plasma levels of sRAGE are higher in symptomatic than asymptomatic carotid atherosclerosis. Higher levels of sRAGE in symptomatic patients may be markers of a higher degree of vascular inflammation in such patients. PMID- 19811527 TI - MMP/ TIMP balance is modulated in vitro by 15dPGJ(2) in fetuses and placentas from diabetic rats. AB - BACKGROUND: Maternal diabetes is associated with morphological placental abnormalities and foeto-placental impairments. These alterations are linked with a dysregulation of the activity of matrix metalloproteinases (MMPs). We investigated the action of 15deoxyDelta(12,14) prostaglandin J(2) (15dPGJ(2)), a natural ligand of the peroxisome proliferator activated receptor (PPAR) gamma, on MMP-2 and MMP-9 activities and tissue inhibitors of matrix metalloproteinases (TIMP) levels in foetuses and placentas from diabetic rats. MATERIALS AND METHODS: Diabetes was induced in rat neonates by a single streptozotocin administration (90 mg kg(-1) s.c.). At 13.5 days of gestation, foetal and placental homogenates were prepared for the determination of PPARgamma levels (western blot) and 15dPGJ(2) concentration (enzyme-immunoassay), whereas the in vitro effect of 15dPGJ(2) (2 microM) was evaluated on placental and foetal MMPs and TIMP activities (zymography and reverse zymography), nitrate/nitrite concentrations (Griess method) and thiobarbituric acid reactive substances (TBARS). RESULTS: PPARgamma was increased while 15dPGJ(2) was decreased in placentas and foetuses from diabetic rats. 15dPGJ(2) additions were able to reduce the high activities of MMP-2 and MMP-9 present in diabetic placental tissues. 15dPGJ(2) additions reduced MMP-2 activity in control and diabetic foetuses. TIMP-3 levels were decreased in diabetic placentas and 15dPGJ(2) was able to enhance them to control values. Nitrates/nitrites and TBARS, metabolites of MMPs activators, were increased in the diabetic placenta and reduced by 15dPGJ(2). CONCLUSIONS: This study demonstrates that 15dPGJ(2) is a potent modulator of the balance between MMP activities and TIMP levels, which is needed in the correct formation and function of the placenta and foetal organs. PMID- 19811528 TI - Intrinsic activity and positive feedback in motor circuits in organotypic spinal cord slice cultures. AB - In co-cultures of embryonic rat spinal cord slices and skeletal muscle, spinal motoneurons innervate muscle fibres and drive muscle contractions. However, multi electrode array (MEA) recordings show that muscle contractions often appear in the absence of population activity in the spinal cord networks. Such uncorrelated muscle activity persists when the population bursts in the neuronal networks are prevented by un-coupling the network with the glutamatergic antagonists CNQX and D-APV. By contrast, the uncorrelated muscle activity is fully suppressed by the muscular nicotinic antagonist D-tubocurarine. Together, these findings confirm the previous finding that motoneurons drive muscle fibres in this preparation and suggest that they are intrinsically spiking in the absence of synaptic input. Intracellular recordings from spinal neurons support this suggestion. Analysing the correlated muscle activity, we found that in 15% of the population bursts, muscle activity appears at the beginning or before neuronal activity, suggesting that in these cases motoneurons initiate the population activity. Both the total number of population bursts and the percentage of such bursts that are initiated by muscle activity are reduced by a block of nicotinic receptors. Uncorrelated muscle and neuronal activity is reduced by the gap junction blocker carbenoxolone, suggesting that electrical coupling is involved in the generation of this activity. Together, these findings suggest that intrinsic firing of motoneurons may contribute to the activation of population bursts through cholinergic positive feedback loops in cultured spinal networks. PMID- 19811529 TI - IL1RAPL1 controls inhibitory networks during cerebellar development in mice. AB - Abnormalities in the formation and function of cerebellar circuitry potentially contribute to cognitive deficits in humans. In the adult, the activity of the sole output neurons of the cerebellar cortex - the Purkinje cells (PCs) - is shaped by the balance of activity between local excitatory and inhibitory circuits. However, how this balance is established during development remains poorly understood. Here, we investigate the role of interleukin-1 receptor accessory protein-like 1 (IL1RAPL1), a protein linked to cognitive function which interacts with neuronal calcium sensor 1 (NCS-1) in the development of mouse cerebellum. Using Il1rapl1-deficient mice, we found that absence of IL1RAPL1 causes a transient disinhibition of deep cerebellar nuclei neurons between postnatal days 10 and 14 (P10/P14). Upstream, in the cerebellar cortex, we found developmental perturbations in the activity level of molecular layer interneurons (MLIs), resulting in the premature appearance of giant GABAA-mediated inhibitory post-synaptic currents capable of silencing PCs. Examination of feed-forward recruitment of MLIs by parallel fibres shows that during this P10/P14 time window, MLIs were more responsive to incoming excitatory drive. Thus, we conclude that IL1RAPL1 exerts a key function during cerebellar development in establishing local excitation/inhibition balance. PMID- 19811530 TI - Hypocretin/orexin contributes to the expression of some but not all forms of stress and arousal. AB - Hypocretin/orexin has a well-established role in wakefulness and in the maintenance of arousal. Because stress is associated with arousal, it has been proposed that hypocretin is also involved in stress. However, it is not clear if this is true for all forms of stress. To clarify this issue, we compared four conditions combining high arousal with no or low stress (wakefulness and exploration) or high stress (contextual fear and restraint) in the rat. We looked at Fos expression in hypocretin neurons, hypocretin-1 levels in cerebrospinal fluid and cardiovascular and behavioural changes after pharmacological blockade with the dual hypocretin receptor antagonist, almorexant. Fos expression in hypocretin neurons was highest with wakefulness and exploration, also high with fear but not significant with restraint. Hypocretin-1 levels were consistent with this pattern, although the differences were not as marked. Hypocretin receptor blockade with almorexant reduced the pressor, tachycardic and locomotor responses of wakefulness and exploration as well as the pressor and sympathetic component of the tachycardic response of fear. In contrast, almorexant did not reduce the pressor and tachycardic responses of restraint and nor did it reduce the pressor, tachycardic and locomotor responses of another stressor, i.e. cold exposure. Thus, hypocretin is not involved in all forms of stress. Comparison of the different conditions suggests that, regardless of stress, hypocretin involvement occurs when the arousal associated with the response includes increased attention to environmental cues. When it does, hypocretin will at least contribute to the cardiovascular response. The findings are of clinical relevance to some forms of psychological stress. PMID- 19811531 TI - Fear conditioning enhances spontaneous AMPA receptor-mediated synaptic transmission in mouse hippocampal CA1 area. AB - AMPA receptor-mediated synaptic modifications in the amygdala have been reported to sustain cued fear conditioning. However, the hippocampal formation is also critically involved in fear learning. Therefore, we examined whether fear conditioning is also accompanied by changes in AMPA receptor-mediated synaptic transmission in the hippocampus. We focused on spontaneous miniature excitatory post-synaptic currents (mEPSCs). Young adult mice were trained using tone/footshock pairings and contextual/cued memories were tested 3-4 h and 1 day later. We found that the mEPSC frequency was significantly enhanced when recorded 3 h, but not 24 h, after fear conditioning training. Fear training induced a slight enhancement in the mEPSC amplitude at 3 h after training. The increased mEPSC frequency and amplitude were absent in animals that were only exposed to footshock or novelty or unpaired tone/footshock training. This implies that learning the association between context, tone and footshock transiently enhances hippocampal CA1 spontaneous synaptic transmission, which may contribute to the encoding of the fearful event. PMID- 19811532 TI - Fasting biases brain reward systems towards high-calorie foods. AB - Nutritional state (e.g. fasted vs. fed) and different food stimuli (e.g. high calorie vs. low-calorie, or appetizing vs. bland foods) are both recognized to change activity in brain reward systems. Using functional magnetic resonance imaging, we have studied the interaction between nutritional state and different food stimuli on brain food reward systems. We examined how blood oxygen level dependent activity within a priori regions of interest varied while viewing pictures of high-calorie and low-calorie foods. Pictures of non-food household objects were included as control stimuli. During scanning, subjects rated the appeal of each picture. Twenty non-obese healthy adults [body mass index 22.1 +/- 0.5 kg/m(2) (mean +/- SEM), age range 19-35 years, 10 male] were scanned on two separate mornings between 11:00 and 12:00 h, once after eating a filling breakfast ('fed': 1.6 +/- 0.1 h since breakfast), and once after an overnight fast but skipping breakfast ('fasted': 15.9 +/- 0.3 h since supper) in a randomized cross-over design. Fasting selectively increased activation to pictures of high-calorie over low-calorie foods in the ventral striatum, amygdala, anterior insula, and medial and lateral orbitofrontal cortex (OFC). Furthermore, fasting enhanced the subjective appeal of high-calorie more than low calorie foods, and the change in appeal bias towards high-calorie foods was positively correlated with medial and lateral OFC activation. These results demonstrate an interaction between homeostatic and hedonic aspects of feeding behaviour, with fasting biasing brain reward systems towards high-calorie foods. PMID- 19811533 TI - Tissue-type plasminogen activator-plasmin-BDNF modulate glutamate-induced phase shifts of the mouse suprachiasmatic circadian clock in vitro. AB - The mammalian circadian clock in the suprachiasmatic nucleus (SCN) maintains environmental synchrony through light signals transmitted by glutamate released from retinal ganglion terminals. Brain-derived neurotrophic factor (BDNF) is required for light/glutamate to reset the clock. In the hippocampus, BDNF is activated by the extracellular protease, plasmin, which is produced from plasminogen by tissue-type plasminogen activator (tPA). We provide data showing expression of proteins from the plasminogen activation cascade in the SCN and their involvement in circadian clock phase-resetting. Early night glutamate application to SCN-containing brain slices resets the circadian clock. Plasminogen activator inhibitor-1 (PAI-1) blocked these shifts in slices from wild-type mice but not mice lacking its stabilizing protein, vitronectin (VN). Plasmin, but not plasminogen, prevented inhibition by PAI-1. Both plasmin and active BDNF reversed alpha(2)-antiplasmin inhibition of glutamate-induced shifts. alpha(2)-Antiplasmin decreased the conversion of inactive to active BDNF in the SCN. Finally, both tPA and BDNF allowed daytime glutamate-induced phase resetting. Together, these data are the first to demonstrate expression of these proteases in the SCN, their involvement in modulating photic phase-shifts, and their activation of BDNF in the SCN, a potential 'gating' mechanism for photic phase-resetting. These data also demonstrate a functional interaction between PAI 1 and VN in adult brain. Given the usual association of these proteins with the extracellular matrix, these data suggest new lines of investigation into the locations and processes modulating mammalian circadian clock phase-resetting. PMID- 19811534 TI - IQ-ArfGEF/BRAG1 is associated with synaptic ribbons in the mouse retina. AB - IQ-ArfGEF/BRAG1 is a guanine nucleotide exchange factor for ADP ribosylation factors (Arfs), which are implicated in membrane trafficking and actin cytoskeleton dynamics. In this study, we examined the immunohistochemical localization of IQ-ArfGEF/BRAG1 in the adult mouse retina using light and electron microscopy. IQ-ArfGEF/BRAG1 was distributed in a punctate manner and colocalized well with RIBEYE in both the outer and inner plexiform layers. Immunoelectron microscopic analysis showed that IQ-ArfGEF/BRAG1 was localized at the synaptic ribbons of photoreceptors. When heterologously expressed in HeLa cells, IQ-ArfGEF/BRAG1 was recruited to RIBEYE-containing clusters and formed an immunoprecipitable complex with RIBEYE. Furthermore, immunoprecipitation analysis showed that anti-IQ-ArfGEF/BRAG1 antibody efficiently pulled down RIBEYE from retinal lysates. These findings indicate that IQ-ArfGEF/BRAG1 is a novel component of retinal synaptic ribbons and forms a protein complex with RIBEYE. PMID- 19811535 TI - Numerical discrimination in newborn infants as revealed by event-related potentials to tone sequences. AB - Humans are able to attentively discriminate number from 6 months of age. However, the age of the emergence of this ability at the pre-attentive stage of processing remains unclear. Event-related potentials (ERPs) were recorded in newborn human infants aged from 3 to 5 days. At 500-ms intervals, the infants were passively exposed to 200-ms sequences of four tones. Each tone could be either 1000 or 1500 Hz in frequency. In most sequences (standards), the ratio of the tones of one frequency to those of the other frequency in a sequence was 2 : 2. In the remaining sequences (deviants, P = 0.1), this ratio was either 3 : 1 or 4 : 0. The mismatch response of ERPs could not be found for 3 : 1 deviants, but it was a robust finding for 4 : 0 deviants, showing the neurophysiological ability of the infants to register the larger deviant-standard difference. The findings suggest very early sensitivity to auditory numerical information in infancy. PMID- 19811536 TI - Daily rhythms and sex differences in vasoactive intestinal polypeptide, VIPR2 receptor and arginine vasopressin mRNA in the suprachiasmatic nucleus of a diurnal rodent, Arvicanthis niloticus. AB - Diurnal and nocturnal animals differ with respect to the time of day at which the ovulatory surge in luteinizing hormone occurs. In some species this is regulated by the suprachiasmatic nucleus (SCN), the primary circadian clock, via cells that contain vasoactive intestinal polypeptide (VIP) and vasopressin (AVP). Here, we evaluated the hypothesis that chronotype differences in the timing of the luteinizing hormone surge are associated with rhythms in expression of the genes that encode these neuropeptides. Diurnal grass rats (Arvicanthis niloticus) were housed in a 12/12-h light-dark cycle and killed at one of six times of day (Zeitgeber time 1, 5, 9, 13, 17, 21; ZT 0 = lights-on). In-situ hybridization was used to compare levels of vip, avp and VIP receptor mRNA (vipr2) in the SCN of intact females, ovariectomized females, ovariectomized females given estradiol and intact males. We found a sex difference in vip rhythms with a peak occurring at ZT 13 in males and ZT 5 in intact females. In all groups avp mRNA rhythms peaked during the day, from ZT 5 to ZT 9, and had a trough in the dark at ZT 21. There was a modest rhythm and sex difference in the pattern of vipr2. Most importantly, the patterns of each of these SCN rhythms relative to the light-dark cycle resembled those seen in nocturnal rodents. Chronotype differences in timing of neuroendocrine events associated with ovulation are thus likely to be generated downstream of the SCN. PMID- 19811537 TI - Effect of siRNA terminal mismatches on TRBP and Dicer binding and silencing efficacy. AB - To enhance silencing and avoid off-target effects, siRNAs are often designed with an intentional bias to ensure that the end of the siRNA that contains the guide strand 5' end is less stably hybridized relative to the end containing the passenger strand 5' end. One means by which this is accomplished is to introduce a terminal mismatch, typically by changing the passenger strand sequence to impair its hybridization with the guide strand 5' end. However, there are conflicting reports about the influence of terminal mismatches on the silencing efficacy of siRNAs. Here, the silencing efficiency of siRNAs with a terminal mismatch generated either by altering the guide strand (at the 5' end, nucleotide 1) or the passenger strand (nucleotide 19 from the 5' end) was examined. Subsequently, we studied the relationship between the silencing efficiency of the siRNAs and their binding to the RNA-induced silencing complex loading complex proteins HIV transactivating response RNA-binding protein and Dicer in H1299 cytoplasmic extracts. Binding of siRNA and the transactivating response RNA binding protein was significantly reduced by terminal mismatches, which largely agrees with the reduction in eventual silencing efficacy of the siRNAs. Single terminal mismatches led to a small increase in Dicer binding, as expected, but this did not lead to an improvement in silencing activity. These results demonstrate that introduction of mismatches to control siRNA asymmetry may not always improve target silencing, and that care should be taken when designing siRNAs using this technique. PMID- 19811538 TI - Phylogenetic and metagenomic analysis of Verrucomicrobia in former agricultural grassland soil. AB - The bacterial phylum Verrucomicrobia has a widespread distribution, and is known to be one of the most common and diverse phyla in soil habitats. However, members of this phylum have typically been recalcitrant to cultivation methods, hampering the study of this presumably important bacterial group. In this study, we examine the phylogenetic diversity of the Verrucomicrobia in a former agricultural field and gain access to genomic information via a metagenomic approach. We examined Verrucomicrobia-like 16S rRNA gene sequences recovered from general bacterial and phylum-specific libraries, revealing a dominance of subdivisions 1 and 2. A PCR based screening method was developed to identify inserts containing verrucomicrobial 16S rRNA genes within a large-insert metagenomic library, and on screening of 28,800 clones, four fosmids were identified as containing verrucomicrobial genomic DNA. Full-length sequencing of fosmid inserts and gene annotation identified a total of 98 ORFs, representing a range of functions. No conservation of gene order was observed adjacent to the ribosomal operons. Fosmid inserts were further analyzed for tetranucleotide frequencies to identify remnants of past horizontal gene transfer events. The metagenomic approach utilized proved to be suitable for the recovery of verrucomicrobial genomic DNA, thereby providing a window into the genomes of members of this important, yet poorly characterized, bacterial phylum. PMID- 19811539 TI - Links between methanotroph community composition and CH oxidation in a pine forest soil. AB - The main gap in our knowledge about what determines the rate of CH(4) oxidation in forest soils is the biology of the microorganisms involved, the identity of which remains unclear. In this study, we used stable-isotope probing (SIP) following (13)CH(4) incorporation into phospholipid fatty acids (PLFAs) and DNA/RNA, and sequencing of methane mono-oxygenase (pmoA) genes, to identify the influence of variation in community composition on CH(4) oxidation rates. The rates of (13)C incorporation into PLFAs differed between horizons, with low (13)C incorporation in the organic soil and relatively high (13)C incorporation into the two mineral horizons. The microbial community composition of the methanotrophs incorporating the (13)C label also differed between horizons, and statistical analyses suggested that the methanotroph community composition was a major cause of variation in CH(4) oxidation rates. Both PLFA and pmoA-based data indicated that CH(4) oxidizers in this soil belong to the uncultivated 'upland soil cluster alpha'. CH(4) oxidation potential exhibited the opposite pattern to (13)C incorporation, suggesting that CH(4) oxidation potential assays may correlate poorly with in situ oxidation rates. The DNA/RNA-SIP assay was not successful, most likely due to insufficient (13)C-incorporation into DNA/RNA. The limitations of the technique are briefly discussed. PMID- 19811540 TI - Programmed cell death and the pathogenesis of tissue injury induced by type A Francisella tularensis. AB - Francisella tularensis is a highly virulent bacterial species that causes various forms of tularemia in humans. The urgency in understanding the pathogenesis of these diseases has stimulated unprecedented interest in this bacterial species over the past few years. Recent findings underscore a number of important distinctions between the Francisella ssp. and emphasize the importance of using type A F. tularensis strains when characterizing pathophysiological responses that are relevant to the lethal forms of human disease. This review focuses on the mediators of cell death induction in infected tissues and the implications of these processes on the pathophysiological changes observed in various host species. PMID- 19811541 TI - Unusual binding properties of the dockerin module of Clostridium thermocellum endoglucanase CelJ (Cel9D-Cel44A). AB - Cellulosomes are cellulolytic complexes produced by anaerobic bacteria, and are composed of a scaffolding protein and several catalytic components. The complexes are formed by highly specific interactions of one of the reiterated cohesin modules of the scaffolding protein with a dockerin module of the catalytic components. The affinities of a dockerin module of Clostridium thermocellum CelJ (Cel9D-Cel44A) for several cohesin modules from C. thermocellum and Clostridium josui scaffolding proteins were quantitatively measured by surface plasmon resonance analysis. The recombinant CelJ dockerin-containing protein interacted with three recombinant C. josui cohesin proteins as well as recombinant C. thermocellum cohesin proteins beyond the so-called 'species specificity' of the dockerin and cohesin interactions. However, this protein did not recognize a second cohesin module from the C. josui scaffolding protein, suggesting that the catalytic components are not necessarily arranged randomly on a scaffolding protein in native cellulosomes. PMID- 19811542 TI - 2-Methylhopanoids are maximally produced in akinetes of Nostoc punctiforme: geobiological implications. AB - 2-Methylhopanes, molecular fossils of 2-methylbacteriohopanepolyol (2-MeBHP) lipids, have been proposed as biomarkers for cyanobacteria, and by extension, oxygenic photosynthesis. However, the robustness of this interpretation is unclear, as 2-methylhopanoids occur in organisms besides cyanobacteria and their physiological functions are unknown. As a first step toward understanding the role of 2-MeBHP in cyanobacteria, we examined the expression and intercellular localization of hopanoids in the three cell types of Nostoc punctiforme: vegetative cells, akinetes, and heterocysts. Cultures in which N. punctiforme had differentiated into akinetes contained approximately 10-fold higher concentrations of 2-methylhopanoids than did cultures that contained only vegetative cells. In contrast, 2-methylhopanoids were only present at very low concentrations in heterocysts. Hopanoid production initially increased threefold in cells starved of nitrogen but returned to levels consistent with vegetative cells within 2 weeks. Vegetative and akinete cell types were separated into cytoplasmic, thylakoid, and outer membrane fractions; the increase in hopanoid expression observed in akinetes was due to a 34-fold enrichment of hopanoid content in their outer membrane relative to vegetative cells. Akinetes formed in response either to low light or phosphorus limitation, exhibited the same 2 methylhopanoid localization and concentration, demonstrating that 2 methylhopanoids are associated with the akinete cell type per se. Because akinetes are resting cells that are not photosynthetically active, 2 methylhopanoids cannot be functionally linked to oxygenic photosynthesis in N. punctiforme. PMID- 19811543 TI - Efficacy and safety of highly purified, doubly virus-inactivated VWF/FVIII concentrates in inherited von Willebrand's disease: results of an Italian cohort study on 120 patients characterized by bleeding severity score. AB - The efficacy of highly purified VWF/FVIII concentrates with standardized ristocetin cofactor content (VWF:RCo) has been already proven in patients with von Willebrand's disease (VWD). Aim of this retrospective study is to confirm efficacy and safety of two highly purified, doubly virus-inactivated VWF/FVIII concentrates in a large cohort of patients with VWD who were characterized at enrolment by bleeding severity score. Study drugs Alphanate or Fanhdi were given to 120 cases (51 males, 69 females, median age 50 years, range 6-83 years). Patients had VWD3 (10), VWD2A (19), VWD2B (25), VWD2M (10) and DDAVP-unresponsive VWD1 (56) and a median bleeding severity score of 8 (range 0-27). A total of 114 bleeding episodes in 55 cases and 131 surgical procedures in 85 cases could be analysed. Excellent-good clinical responses were seen in 97% of bleeding episodes and in 99% of surgical procedures. To prevent recurrent gastrointestinal (GI) bleeding, cerebral (CNS) haemorrhage, haemarthroses, urogenital or multisite bleeding in more severe patients, secondary prophylaxis was also carried out in 15 cases with VWD3 (3), VWD2A (3), VWD2B (2), VWD1 (7). A median dose of 42 IU VWF:RCo kg(-1) given every other day or twice a week over a median period of 334 days (range 24-799) prevented bleeding completely in 13 cases and reduced its incidence in the remaining two. These results confirm the efficacy and safety of the study concentrates, not only in the management of bleeding and surgery but also in secondary prophylaxis of severe VWD. PMID- 19811544 TI - Commentary on Knight et al.: A systematic review of the cost-effectiveness of rFVIIa and APCC in the treatment of minor/moderate bleeding episodes for haemophilia patients with inhibitors. PMID- 19811545 TI - Patient factors in the implementation of decision aids in general practice: a qualitative study. AB - BACKGROUND: Decision aids (DAs) have been developed to help patients make treatment decisions. Research shows that they are effective in increasing patients' knowledge of treatment options without raising anxiety or conflict. However, they have not been routinely adopted for use in general practice in the UK and there are few reports addressing strategies to introduce them. OBJECTIVE: To examine patients' views about a variety of DAs for different conditions (heart disease, osteoporosis, osteoarthritis and breast cancer) in order to inform a strategy to introduce them into general practice. SETTING AND PARTICIPANTS: General practice patients over the age of 18 years being or having been treated for one of the conditions above. METHODS: Qualitative study involving 12 focus groups with 77 patients evaluating decision aids relevant to their conditions. A semi-structured interview guide was used to generate discussions about the applicability of the DAs in routine general practice. RESULTS: Patients welcomed DAs for their educational and informational content. Reactions to the DAs were influenced by patients' own personal desires for involvement. The main concerns were that the use of DAs would potentially shift the onus of decision making responsibility on to the patient and about the practical challenges to implementation. CONCLUSIONS: Clinicians will need to make explicit to patients that DAs are an adjunct to routine care and not a replacement, and therefore do not represent a derogation of responsibility. DAs need to be used as an integral part of the communication and support process for patients who want them. PMID- 19811546 TI - Sperm characteristics and teratology in rats following vas deferens occlusion with RISUG and its reversal. AB - The functional success of the reversal of vas occlusion by styrene maleic anhydride (RISUG), using the solvent vehicle, Dimethyl Sulphoxide (DMSO), has been investigated. Reversal with DMSO was carried out in Wistar albino rats 90 days after bilateral vas occlusion. The body weight, organ weight, sperm characteristics, fertility test and teratology, including skeletal morphology were evaluated in vas occlusion and reversal animals and in F(1) progenies to assess the functional success of the occlusion and reversal. Body weight, organ weight and the cauda epididymal sperm characteristics of vas occlusion and reversal animals and of F(1) progenies were comparable to control. Ejaculated spermatozoa in the vaginal smear showed detached head/tail, acrosomal damage, bent midpiece, bent tail and morphological aberrations in sperm head after vas occlusion, which returned to normal, 90 days after reversal. Monthly fertility test, post-injection showed 0% fertility, which improved gradually and 100% fertility was achieved 90 days after reversal. The fertility/pregnancy/implantation record and skeletal morphology of the offspring were comparable to control. The results suggest functional success and safety of vas occlusion reversal by DMSO. PMID- 19811547 TI - Clinical guideline for male lower urinary tract symptoms. AB - This article is a shortened version of the clinical guideline for lower urinary tract symptoms (LUTS), which has been developed in Japan for symptomatic men aged 50 years and over irrespective of presumed diagnoses. The guideline was formed on the PubMed database between 1995 and 2007 and other relevant sources. The causes of male LUTS are diverse and attributable to diseases/dysfunctions of the lower urinary tract, prostate, nervous system, and other organ systems, with benign prostatic hyperplasia, bladder dysfunction, polyuria, and their combination being most common. The mandatory assessment should comprise medical history, physical examination, urinalysis, and measurement of serum prostate-specific antigen. Symptom and quality of life questionnaires, bladder diary, residual urine measurement, urine cytology, urine culture, measurement of serum creatinine, and urinary tract ultrasonography would be optional tests. The Core Lower Urinary Tract Symptom Score Questionnaire may be useful in quickly capturing important symptoms. Severe symptoms, pain symptoms, and other clinical problems would indicate urological referral. One should be careful not to overlook underlying diseases such as infection or malignancy. The treatment should be initiated with conservative therapy and/or medicine such as alpha(1)-blockers. Treatment with anticholinergic agents should be reserved only for urologists, considering the risk of urinary retention. The present guideline should help urologists and especially non-urologists treat men with LUTS. PMID- 19811548 TI - Angiogenesis in renal cell carcinoma: the role of tumor-associated macrophages. AB - OBJECTIVE: To explore vascularity and associated molecules in renal cell carcinoma (RCC) and to study their correlations to disease outcome. METHODS: Tissue samples from 51 Japanese patients with renal cell carcinoma (RCC) were obtained between November 1997 and August 2000. Pyrimidine nucleoside phosphorylase and vascular endothelial growth factor (VEGF) levels of RCC and normal kidney tissue were determined by enzyme-linked immunosorbent assay. Microvessel density (MVD) was measured by immunohistochemistry using anti-factor VIII-related antigen and CD34. The number of infiltrating tumor-associated macrophages (TAM) was measured by immunohistochemistry using anti-CD68 antibody. RESULTS: Pyrimidine nucleoside phosphorylase and VEGF levels were significantly higher in RCC than in normal kidney tissue. The VEGF level was higher in more progressive (high grade, larger or symptomatic) RCC. Although MVD as determined by the factor VIII level was higher in larger tumors, MVD determined by CD34 was higher in low-grade and low-stage tumors. Patients with symptoms, large tumor or high stage showed higher numbers of TAM. VEGF level and TAM were significantly higher in patients with recurrence than in those without recurrence. In univariate analysis, VEGF, TAM and CD34 tumor grade and stage were identified as prognostic factors. Moreover, TAM was the only independent prognostic factor by multivariate analysis. Among these parameters, only TAM and MVD as determined by factor VIII showed significant correlations. CONCLUSION: TAM and VEGF are substantially involved in tumor progression of RCC. As the TAM count is well correlated to the MVD, the main mechanism of tumor progression by TAM might be angiogenesis. PMID- 19811549 TI - Simple prophylactic procedure of inguinal hernia after radical retropubic prostatectomy: isolation of the spermatic cord. AB - To reduce the incidence of inguinal hernia (IH) after radical retropubic prostatectomy (RRP), a simple prophylactic procedure was carried out during RRP. A consecutive 82 patients who had undergone RRP for clinically localized prostate cancer between July 2002 and October 2006 at Toride Kyodo General Hospital were enrolled. From July 2002 to November 2003, 20 patients underwent conventional RRP. Thereafter, 62 patients underwent conventional RRP with blunt dissection of the peritoneum at the internal inguinal ring and isolation of the spermatic cord from the peritoneum as a prophylactic procedure for IH. There was no significant difference in patient characteristics between the two groups. In the conventional RRP group, IH occurred in 10 patients during a median range follow-up period of 41 (1 to 73) months. In contrast, in the RRP plus prophylactic procedure group, IH occurred in one patient (1.6%) during a median range follow-up period of 41 (25 to 59) months. The incidence of IH after RRP plus the prophylactic procedure was significantly lower than that after conventional RRP, indicating the efficacy of the presented procedure. PMID- 19811550 TI - A clinical review of infected wound treatment with Vacuum Assisted Closure (V.A.C.) therapy: experience and case series. AB - Over the last decade Vacuum Assisted Closure((R)) (KCI Licensing, Inc., San Antonio, TX) has been established as an effective wound care modality for managing complex acute and chronic wounds. The therapy has been widely adopted by many institutions to treat a variety of wound types. Increasingly, the therapy is being used to manage infected and critically colonized, difficult-to-treat wounds. This growing interest coupled with practitioner uncertainty in using the therapy in the presence of infection prompted the convening of an interprofessional expert advisory panel to determine appropriate use of the different modalities of negative pressure wound therapy (NPWT) as delivered by V.A.C.((R)) Therapy and V.A.C. Instill((R)) with either GranuFoam() or GranuFoam Silver() Dressings. The panel reviewed infected wound treatment methods within the context of evidence-based medicine coupled with experiential insight using V.A.C.((R)) Therapy Systems to manage a variety of infected wounds. The primary objectives of the panel were 1) to exchange state-of-practice evidence, 2) to review and evaluate the strength of existing data, and 3) to develop practice recommendations based on published evidence and clinical experience regarding use of the V.A.C.((R)) Therapy Systems in infected wounds. These recommendations are meant to identify which infected wounds will benefit from the most appropriate V.A.C.((R)) Therapy System modality and provide an infected wound treatment algorithm that may lead to a better understanding of optimal treatment strategies. PMID- 19811551 TI - Translation and validation of a Chinese language version of the Early Childhood Oral Health Impact Scale (ECOHIS). AB - OBJECTIVE: This study aimed to adapt the Early Childhood Oral Health Impact Scale (ECOHIS) for pre-school children in a Chinese speaking community and to investigate its psychometric properties (validity and reliability). METHODS: A Chinese language version of the ECOHIS was derived through a forward-backward translation and tested for face and content validity among a focus group. A convenient sample of pre-school children (n = 111) was recruited (including a sub sample with early childhood caries and caries-free children). Parents of the children self-completed the derived Chinese-ECOHIS measure. Validity of the measure was assessed by investigating the relationship between dental caries status and Chinese-ECOHIS scores (construct and criterion validity). A sub-sample of the parents repeated the ratings of the measure to enable reliability assessments. Both internal and test-retest reliability were determined. RESULTS: A Chinese version of ECOHIS was derived with minor modification to the original version. Chinese-ECOHIS scores were associated with children's caries experience (dmft) (r = 0.66, P < 0.05) supporting convergent validity. In addition, variations in ECOHIS scores were apparent with respect to caries and caries-free groups (P < 0.001), supporting the ability to distinguish between patient groups. Cronbach's alpha values (internal reliability) for total ECOHIS score were 0.91 and intraclass correlation coefficient value (test-retest reliability) was 0.64. CONCLUSIONS: A Chinese version of the ECOHIS was developed and demonstrated acceptable validity and reliability. These findings can enable assessments of pre school child oral health-related quality of life in Chinese speaking communities. PMID- 19811552 TI - Obstructive sleep apnoea--an update. AB - Obstructive sleep apnoea (OSA) is a common disorder characterized by the repetitive complete or partial collapse of the upper airway during sleep. It results in intermittent hypoxaemia and hypercapnia, cortical arousals and surges of sympathetic activity. The occurrence of OSA has also been linked to serious long-term adverse health consequences; such as hypertension, metabolic dysfunction, cardiovascular disease, neurocognitive deficits and motor vehicle accidents. There have been several advances in the field of particular clinical importance: (i) the development of portable monitoring as part of a simplified clinical algorithm for the diagnosis of OSA in selected patients; (ii) growing awareness of the cardio-metabolic health consequences of OSA and (iii) emerging evidence to support a range of non-continuous positive airway pressure (CPAP) treatment modalities, such as oral appliances. PMID- 19811553 TI - A survey of Australian clinical registries: can quality of care be measured? AB - BACKGROUND: Clinical quality registries gather and analyse information to monitor and enhance the quality of care received by patients. The aim of the present study was to determine the attributes of Australian clinical registries to identify their capacity to accurately assess quality of care. METHODS: A survey was distributed to registry custodians managing multi-site clinical outcome registries. They were asked to self-report on general aspects of registries, including coverage, length of operation, data collection process, data management, quality of data and registry governance structures. RESULTS: A total of 28 registries were identified and all provided responses to the survey. The majority of the registries require modifications to their procedures in order to provide useful and reliable information for quality improvement purposes. Thirteen registries (46%) did not assess or recruited fewer than 80% of the eligible population and 23 (82%) did not formally audit reliability of coding at the clinical level. Five (18%) did not collect the information required for basic risk adjustment of outcome measures. While most registries produced reports for providers and interested parties, the approach to disseminating this information was highly variable. CONCLUSION: Clinical registries provide the most credible information about quality of care. However, most key registries in Australia require some adaptation of procedures in order to accomplish this task. Funding should be provided to enable registries to make the necessary changes. PMID- 19811554 TI - Exploratory study of the 'weekend effect' for acute medical admissions to public hospitals in Queensland, Australia. AB - AIMS: To determine whether in-hospital deaths of patients admitted through emergency departments with acute exacerbations of chronic obstructive pulmonary disease (COPD), acute myocardial infarction, intracerebral haemorrhage and acute hip fracture are increased by weekend versus weekday admission (the 'weekend effect'). METHODS: We performed a retrospective analysis of statewide administrative data from public hospitals in Queensland, Australia, during the 2002/2003-2006/2007 financial years. The primary outcome was 30-day in-hospital mortality. The secondary outcome of 2-day in-hospital mortality helped determine whether increased mortality of weekend admissions was closely linked to weekend medical care. RESULTS: During the study period, there were 30 522 COPD, 17 910 acute myocardial infarction, 4183 acute hip fracture and 1781 intracerebral haemorrhage admissions. There was no significant weekend effect on 30-day in hospital mortality for COPD (adjusted risk ratio = 0.92, 95% CI: 0.81-1.04, P= 0.222), intracerebral haemorrhage (adjusted risk ratio = 1.01, 95% CI: 0.86-1.16, P= 0.935) or acute hip fracture (adjusted risk ratio = 0.78, 95% CI: 0.54-1.03, P= 0.13). There was a significant weekend effect for acute myocardial infarction (adjusted risk ratio = 1.15, 95% CI: 1.03-1.26, P= 0.007). Two-day in-hospital mortality showed similar results. CONCLUSION: This is the first Australian study on the 'weekend effect' (in a cohort other than neonates), and the first study worldwide to assess specifically the weekend effect among COPD patients. Observed patterns were consistent with overseas research. There was a significant weekend effect for myocardial infarction. Further research is needed to determine whether location (e.g. rural), clinical (e.g. disease severity) and service provision factors (e.g. access to invasive procedures) influence the weekend effect for acute medical conditions in Australia. PMID- 19811555 TI - What is the role of invasive versus non-invasive coronary angiography in the investigation of patients suspected to have coronary heart disease? AB - The improved technology of multi-slice cardiac CT angiography (CTA) has enabled production of high quality images of the coronary arteries. The sensitivity and specificity of the test in identifying patients with obstructive coronary lesions in 64-slice and later generations of scanners is high. To enable effective use of CTA in the clinical setting, a better understanding of this technology, particularly in comparison to invasive coronary angiography, is needed. In this article we discuss the characteristics of CTA in comparison to invasive coronary angiography and discuss the role of CTA in the diagnosis of coronary artery disease (CAD). Newer CTA scanners and individualized scanning protocols can minimize radiation exposure from CTA. CTA can provide more information than invasive coronary angiography on the type and burden of atherosclerotic plaque in the coronary tree. CTA is most useful in those at intermediate risk of CAD. PMID- 19811556 TI - Emergency department management of exacerbation of chronic obstructive pulmonary disease: audit of compliance with evidence-based guidelines. AB - BACKGROUND: Emergency departments (ED) play a key role in management of exacerbation of chronic obstructive airways disease (COPD). Current guidelines for management of exacerbation of COPD showed highest levels of evidence (Level A and B) were related to use of medications and non-invasive positive pressure ventilation (NIPPV). AIMS: The aim of this study was to examine compliance with high level evidence for management of exacerbation of COPD during the first 4 h of ED care. METHODS: A retrospective medical record audit was conducted at four public and one private ED in Melbourne, Australia. Participants were adult patients with COPD presenting to the ED with a primary complaint of shortness of breath from July 2006 to July 2007. Outcome measures were compliance with evidence-based recommendations regarding use of bronchodilators, methylxanthines, steroids and NIPPV. RESULTS: Of 273 patients in this study, 72.4% received short acting beta-agonist bronchodilators, 37.8% received an inhaled short-acting anticholinergic medication and 56.6% received systemic steroid therapy. NIPPV was used in 21 patients, 15 of whom had documentation of acidosis and/or hypercapnia). CONCLUSIONS: There was variation in the use of high level evidence for the ED management of exacerbation of COPD. The highest rate of compliance was non-use of methylxanthines and the greatest deficit was poor compliance with evidence related to NIPPV. There was also scope for improvement in the use of bronchodilators and systemic steroids. PMID- 19811557 TI - Referral patterns to a palliative care service in rural Australia servicing indigenous Australians. AB - BACKGROUND: Territory palliative care (TPC) of the Northern Territory covers a population of 150000 over 400000 km2 in urban, rural and remote settings. Thirty two per cent of the population are indigenous Australians. There is little documentation of palliative services to the indigenous Australians. METHODS: We retrospectively reviewed the referral characteristics of patients who died in or were discharged from the TPC over 6 months in order to define unique characteristics of palliative care of such a population. RESULTS: The records of 151 consecutive patients were analysed, with a mean age 63 +/- 17 years, 62% male, 59% non-indigenous, 70% urban dwellers and 71% referred for malignancies. Median time to separation was 69 days, with 33% discharged within 30 days, while 28% survived beyond 6 months. Eighty-two per cent of patients died, and of these 52% in the hospice. indigenous patients were more likely to be younger (54 vs 70 years), female (52% vs 29%), living rurally (52% vs 12%) and to die at home (47% vs 11%). CONCLUSION: These data showed major differences from the largest Australian study published so far, based on a population of predominantly non indigenous Australians. This study identifies for the first time how TPC needs to have an encompassing service plan and delivery model to cater for indigenous patients and people in remote locations. PMID- 19811558 TI - Impact of troponin 1 on long-term mortality after emergency orthopaedic surgery in older patients. AB - OBJECTIVES: To determine the association between post-operative troponin rises and longer term (2-year) mortality after emergency orthopaedic surgery in patients over 60 years of age. METHODS: One hundred and two patients were recruited in 2006 and had inpatient troponin 1 measurements. These patients were followed up by a telephone call annually for complications. RESULTS: At 2 years, 29.4% (30/102) of patients had died. Twenty-five patients (25/54 or 49.3%) with a troponin rise were dead at 2 years compared with five patients without a troponin rise (5/48 or 10.4%), which was significantly different P < 0.0001. Patients with a higher troponin level (>0.1 ug/L) were more likely to be dead at 2 years compared with those with a lower level of troponin. However, when adjusted for other comorbidities the association between troponin elevation and death at 2 years did not persist. Using Cox regression multivariate analysis, only one factor, sustaining an in-hospital cardiac event odds ratio 4.3 (95% confidence interval 1.8-10.3, P = 0.001), was associated with 2 years all-cause mortality . Furthermore, patients who sustained a symptomatic troponin rise (P < 0.0001) or asymptomatic troponin rise (P = 0.004) were more likely to have died at 2 years compared with those with no troponin rise. Three factors were significantly associated with a cardiac event during the second year: (i) post-operative troponin rise (P = 0.05); (ii) pre-morbid atrial fibrillation (P = 0.04); and (iii) post-operative renal failure (P < 0.001). CONCLUSION: Elevated post operative troponin levels are predictive of 1-year but not 2-year mortality in older patients undergoing emergency orthopaedic surgery. PMID- 19811559 TI - Use of the 'CURB 65' score in hospital practice. AB - BACKGROUND: The 'CURB 65' score is a simple well-validated tool for the assessment of severity in community acquired pneumonia (CAP). Whether it is used routinely is unknown. The aim of this study was to determine the frequency of use of the score in routine hospital practice and correlate this with clinical decision making and patient outcome. METHODS: Retrospective cohort study of all patients with CAP (n=174) presenting in three winter months. Demographic and clinical outcome data were recorded and comparisons were made between those patients who had score applied on admission with those that did not. A CURB 65 score was assigned to all patients using data from the patient record, and admission decisions were compared. RESULTS: Only 9 (5.2%) CAP patients had the 'CURB 65' score applied at admission. The overall mortality rate was 3.4%. On applying a score to all cases retrospectively, appropriate admission decisions were made for patients with moderate or severe pneumonia and outcome was in accordance with published results. However, 23 (13%) patients age<65 with mild CAP and no comorbidities were admitted in spite of guideline recommendation for community care. CONCLUSIONS: These data demonstrate that clinical decision making in respect of moderate or severe CAP is the same whether or not a pneumonia severity score is applied. However, routine use of the score will identify patients with mild CAP thus potentially reducing unnecessary admission. PMID- 19811560 TI - Prognostic value of trisomy 8 in primary myelodysplastic syndrome. AB - BACKGROUND: According to the International Prognostic Scoring System (IPSS), sole +8 is categorized as intermediate cytogenetic subgroup. But as some myelodysplastic syndrome (MDS) patients with +8 perhaps progress quickly to acute leukaemia and have shorter survival, some reports have suggested that +8 should be categorized into poor risk cytogenetic group. The aim of this study was to clarify the prognostic role of +8 in MDS patients by comparing patients with normal karyotype, 20q- and -7/7q-. METHODS: The consecutive samples of 435 MDS patients in Shanghai were collected by prospective methods and diagnosed according to World Health Organization classification. Cytogenetic analysis was performed using conventional G-banding karyotyping and fluorescence in situ hybridization techniques. Prognosis was estimated by univariate Log-rank method and multivariate Cox proportional hazard models. RESULTS: Of 424 cases completing the cytogenetic analysis, 71 (16.7%) had +8, including 38 patients with sole +8 (9.0%). No significant difference in median survival was observed between patients with sole +8 and that with +8 and one of other abnormalities. The +8 clone size was not linked to survival. The median survival of patients with +8, normal karyotype and complex karyotype was 25months, 38.1months and 5.9months respectively. However, no significant difference was observed between patients with 20q- (21.4months) and -7/7q- (25.8months). Trisomy 8 was an independent prognostic factor by Cox regression model. CONCLUSION: There is no significant difference in prognosis between patients with +8 and patients with 20q- or -7/7q-. Trisomy 8, 20q- and -7/7q- are categorized as intermediate cytogenetic risk according to our primary study. PMID- 19811561 TI - Prevalence of hepatitis B virus infection in non-Hodgkin lymphoma: a systematic review and meta-analysis. AB - BACKGROUND AND AIM: A recent meta-analysis has demonstrated an association between hepatitis C virus and non-Hodgkin lymphoma (NHL). There is also evidence on the association between hepatitis B virus (HBV) and NHL. The aim of this study was to evaluate this evidence using a meta-analytic approach. METHODS: We searched the MEDLINE database from 1962 to 2008 for case-control studies that have reported the association of HBV with NHL. We calculated the odds ratio (OR) and 95% confidence intervals (CI) to assess the prevalence of HBV infection and pooled the results using three different statistical models. RESULTS: Our search yielded 12 studies with 11 studies (3262 NHL patients, 1,523,205 controls) evaluating HBV infection in NHL and one study (3888 HBV-infected individuals, 205,203 controls) that had investigated for NHL in HBV infection. The OR of detecting HBV infection in NHL when compared with the control population was 2.56 (95% CI, 2.24-2.92) by the fixed effects model; 2.61 (95% CI, 2.29-2.98) by the exact method and 2.67 (95% CI, 2.04-3.49) by the random effects model suggesting a high prevalence of HBV carrier state in lymphoma. There was evidence of statistical heterogeneity which disappeared after exclusion of retrospective studies on sensitivity analysis. CONCLUSIONS: The results of this study suggest a possible causal relation between HBV infection and NHL which needs to be confirmed by experimental and epidemiological studies. In countries where prevalence of HBV infection is 1% or more, it may be prudent to screen patients with NHL for occult HBV infection. PMID- 19811562 TI - Lack of association between elevated mean red cell volume and haematological toxicity in patients receiving long-term methotrexate for rheumatoid arthritis. AB - AIMS: It has been suggested that elevated mean red cell volume (MCV) may be a predictor of haematological toxicity in rheumatoid arthritis (RA) patients receiving methotrexate (MTX). We wished to identify whether there was an association between MCV, red cell folate and haematological toxicity in patients on MTX monotherapy for the long-term management of RA. METHODS: Evidence of haematological toxicity was sought by note review of patients recruited in a cross-sectional study of MTX monotherapy in RA. Retrospective data included MCVs from before MTX initiation and after 3 and 6 months of treatment. Data were collected prospectively every 6 months for up to 2 years after enrolment. Any record of cytopenia or the development of haematological malignancy was recorded from commencement of MTX until the present day. Red cell folate concentrations were tested on enrolment to the study. RESULTS: A total of 165 patients was included, 74.5% female, median disease duration 7 years (range 3 months-57 years). The median duration of MTX treatment was 74.9 months (range 10-241 months) giving 1030.2 patient-years of MTX exposure. Twenty-four patients (14.5%) had a MCV > 98 fL on study entry. Evidence of haematological abnormality was found in six patients (3.6%); chronic lymphocytic leukaemia (1), persistent lymphocytosis (1), persistent monocytosis (1) and neutropenia (3). There was no association between red cell folate or MCV and haematological toxicity. CONCLUSION: Neutropenia and pancytopenia are rare side-effects of MTX therapy in this cohort. Elevated MCV or low mean red cell folate does not appear to be associated with haematological malignancy or toxicity in this cohort of patients on long-term MTX therapy. PMID- 19811563 TI - Functional characteristics of the rat jaw muscles: daily muscle activity and fiber type composition. AB - Skeletal muscles have a heterogeneous fiber type composition, which reflects their functional demand. The daily muscle use and the percentage of slow-type fibers have been shown to be positively correlated in skeletal muscles of larger animals but for smaller animals there is no information. The examination of this relationship in adult rats was the purpose of this study. We hypothesized a positive relationship between the percentage of fatigue-resistant fibers in each muscle and its total duration of use per day. Fourteen Wistar strain male rats (410-450 g) were used. A radio-telemetric device was implanted to record muscle activity continuously from the superficial masseter, deep masseter, anterior belly of digastric and anterior temporalis muscles. The degree of daily muscle use was quantified by the total duration of muscle activity per day (duty time) exceeding specified levels of the peak activity (2, 5, 20 and 50%). The fiber type composition of the muscles was examined by the myosin heavy chain content of the fibers by means of immunohistochemical staining. At lower activity levels (exceeding 2 and 5% of the peak activity), the duty time of the anterior belly of digastric muscle was significantly (P < 0.01) longer than those of the other muscles. The anterior belly of digastric muscle also contained the highest percentage of slow-type fibers (type I fiber and hybrid fiber co-expressing myosin heavy chain I + IIA) (ca. 11%; P < 0.05). By regression analysis for all four muscles, an inter-muscular comparison showed a positive relationship between the duty time (exceeding 50% of the peak activity) and the percentage of type IIX fibers (P < 0.05), which demonstrate intermediate physiological properties relative to type IIA and IIB fibers. For the jaw muscles of adult male rats, the variations of fiber type composition and muscle use suggest that the muscle containing the largest amounts of slow-type fibers (the anterior belly of digastric muscle) is mainly involved in low-amplitude activities and that the amount of type IIX fibers is positively related to the generation of large muscle forces, validating our hypothesis. PMID- 19811564 TI - Differential gene expression of bgp and mgp in trabecular and compact bone of Atlantic salmon (Salmo salar L.) vertebrae. AB - The tissue-specific gene expression of the vitamin K-dependent proteins bone gamma-carboxyglutamate-protein (BGP) and matrix gamma-carboxyglutamate-protein (MGP) in Atlantic salmon (Salmo salar L.) was investigated. In previous studies, BGP, the most abundant non-collagenous protein of bone, was almost exclusively associated with bone, whereas the non-structural protein MGP has a more widespread tissue distribution. In-situ hybridization of juvenile Atlantic salmon ( approximately 40 g, fresh water) vertebrae demonstrated expression of bgp and mgp mRNA in osteoblasts lining the trabecular bone, whereas no staining was observed in the compact bone. By separating the trabecular and compact bone of both juvenile ( approximately 40 g, fresh water) and adult ( approximately 1000 g, sea water) Atlantic salmon, we observed that the two vertebral bone compartments displayed different levels of bgp, whereas no such differences were seen for mgp. Measurements of the mineral content and Ca/P molar ratio in adult salmon revealed no significant differences between trabecular and compact bone. In conclusion, the osteoblasts covering the salmon vertebrae have unique gene expression patterns and levels of bgp and mgp. Further, the study confirms the presence of mRNA from the vitamin K-dependent proteins BGP and MGP in the vertebrae, fin and gills of Atlantic salmon. PMID- 19811565 TI - The diagnosis of the cause of the death of Venerina. AB - Venerina (little Venus) is the name given to a wax model representing a pregnant young woman that was created in Florence (Italy) by Clemente Susini (1754-1814) in 1782. It is currently located in the historic Science Museum of the University of Bologna. The model was constructed so as to enable removal of the thoracic and abdominal walls and various organs, exposing the heart, diaphragm and an opened uterus with a well-developed fetus. The woman is small, about 145 cm (4' 9') tall and of delicate build; she looks like a teenage girl. We know that Clemente Susini worked directly with the cadaver and copied the anatomical preparation exactly. This artist often represented the true structure using a wax mould; the existence of two other versions of this specimen suggests that this model was made in this way. Therefore, Venerina's body may be a faithful representation of a young woman who died while pregnant. Observation of the body confirms that the organs are normal, except for the heart and great vessels. The walls of both ventricles are of equal thickness and the ventricles themselves of approximately equal size. The arch of the aorta and the enlarged pulmonary trunk are connected by a short duct about 3.5 mm in diameter. If this structure represents an open arterial duct, we can deduce that the two ventricles worked under the same conditions of blood pressure, hence their equal wall thickness. If the young woman died from this congenital disease, the cause of death has been diagnosed on a wax model of her body after more than two centuries. PMID- 19811566 TI - Mechanism of skull suture maintenance and interdigitation. AB - Skull sutures serve as growth centers whose function involves multiple molecular pathways. During periods of brain growth the sutures remain thin and straight, later developing complex fractal interdigitations that provide interlocking strength. The nature of the relationship between the molecular interactions and suture pattern formation is not understood. Here we show that by classifying the molecules involved into two groups, stabilizing factors and substrate molecules, complex molecular networks can be modeled by a simple two-species reaction diffusion model that recapitulates all the known behavior of suture pattern formation. This model reproduces the maintenance of thin sutural tissue at early stages, the later modification of the straight suture to form osseous interdigitations, and the formation of fractal structures. Predictions from the model are in good agreement with experimental observations, indicating that the model captures the essential nature of the interdigitation process. PMID- 19811567 TI - Development of a quantitative Real-Time TaqMan PCR assay for determination of the minimal dose of Mycoplasma hyopneumoniae strain 116 required to induce pneumonia in SPF pigs. AB - AIMS: A triplex real-time PCR assay to quantify Mycoplasma hyopneumoniae in specimens from live and dead pigs was developed and validated. The minimal dose of Myc. hyopneumoniae required to induce pneumonia in specific pathogen-free pigs was determined. METHODS AND RESULTS: This TaqMan test simultaneously detected three genes encoding the proteins P46, P97 and P102. All Myc. hyopneumoniae strains analysed were detected, including strains isolated in three countries (France, England and Switzerland) and from several pig farms (n = 33), and the test was specific. The estimated detection thresholds were 1.3 genome equivalents (microl(-1)) for the targets defined in p97 and p102 genes and 13 genome equivalents (microl(-1)) for the segment defined in the p46 gene. This test was used to quantify Myc. hyopneumoniae in specimens sampled from experimentally infected pigs. In live pigs, c. 10(7), 10(8) and 10(10) genome equivalents (ml( 1)) of Myc. hyopneumoniae were detected in the nasal cavities, tonsils and trachea samples, respectively. In dead pigs, 10(8)-10(10) genome equivalents (ml( 1)) of Myc. hyopneumoniae were detected in the lung tissue with pneumonia. The estimated minimal dose of Myc. hyopneumoniae required to induce pneumonia was 10(5) colour-changing units (CCU) per pig (corresponding to 10(8) mycoplasmas). CONCLUSION: The triplex RT-PCR test was validated and can be used for testing samples taken on the pig farms. SIGNIFICANCE AND IMPACT OF THE STUDY: This test should be a very useful tool in pig herds to control enzootic pneumonia or healthy carrier pigs and to study the dynamics of Myc. hyopneumoniae infections. PMID- 19811568 TI - Typing of Listeria monocytogenes strains isolated in Italy by inlA gene characterization and evaluation of a new cost-effective approach to antisera selection for serotyping. AB - AIMS: In this study, 105 Listeria monocytogenes strains isolated from humans, foods and environmental samples were characterized using several typing methods. Moreover, serotyping procedure was evaluated, and a cost-effective methodological approach based on preliminary PCRs screening was proposed. METHODS AND RESULTS: The isolates were analysed by conventional serotyping, multiplex-PCRs for serogroup and lineage identification and PCR-RFLP of inlA gene to identify potentially noninvasive L. monocytogenes. Among the strains, only the serotypes 1/2a, 1/2c, 1/2b, 4b and 3a were identified. The isolates were classified into serogroups I (58.10%), II (22.85%), III (12.38%) and IV (6.67%). Among clinical strains, lineage I was more represented (68.75%) than lineage II; whereas, lineage II was more associated with food (90.24%) and environmental (85.72%) isolates. Most of food (89.02%) and environmental (85.71%) isolates were classified into truncated InlA profiles, whereas the 93.75% of clinical strains were associated with a complete form of the protein. CONCLUSION: Molecular techniques were sensitive and specific for classifying strains into serogroup and lineage and in agreement with the serotyping. Moreover, a preliminary PCRs-based screening was proposed to select only the necessary antisera by a flow chart; this methodological approach allows cost saving up to 42%. Our results further suggest the role of InlA protein in human listeriosis, particularly in immunocompetent individuals, and a correlation between truncated protein and serotype. SIGNIFICANCE AND IMPACT OF THE STUDY: This study further validates molecular methods for L. monocytogenes analysis and proposed a new cost-effective approach for serotyping. It could help to improve a national surveillance network for L. monocytogenes infections in Italy. PMID- 19811569 TI - Antimicrobial activity of honey from the stingless bee Trigona carbonaria determined by agar diffusion, agar dilution, broth microdilution and time-kill methodology. AB - AIMS: The aim of this study was to determine the spectrum of antimicrobial activity of 11 samples of stingless bee honey compared to medicinal, table and artificial honeys. METHODS AND RESULTS: Activity was assessed by agar diffusion, agar dilution, broth microdilution and time-kill viability assays. By agar dilution, minimum inhibitory concentration (MIC) ranges were 4% to >10% (w/v) for Gram-positive bacteria, 6% to >16% (w/v) for Gram-negative bacteria and 6% to >10% (w/v) for Candida spp. By broth microdilution, all organisms with the exception of Candida albicans and Candida glabrata were inhibited at 3 log for Pseudomonas aeruginosa and <1 log for C. albicans. Similar treatment with each control honey resulted in decreases of <1 log for all organisms. CONCLUSIONS: Stingless bee honey has broad-spectrum antibacterial activity although activity against Candida was limited. Stingless bee honey samples varied in activity and the basis for this remains to be determined. SIGNIFICANCE AND IMPACT OF THE STUDY: Stingless bee honey had similar activity to medicinal honey and may therefore have a role as a medicinal agent. PMID- 19811570 TI - In vitro studies as supporting evidence on the efficacy of antimicrobials for mastitis treatment. PMID- 19811572 TI - New protocols for the extraction of nucleic acids from soil. PMID- 19811573 TI - Octreotide for management of chylothorax following bidirectional glenn in a three month-old infant. Comment. PMID- 19811574 TI - Heterogeneous double cardiac tumor: myxoma concomitant with papillary fibroelastoma. AB - Heterogeneous double cardiac tumor is rarely encountered. We present a case of simultaneous resection of myxoma and papillary fibroelastoma. Transesophageal echocardiography showed typical characteristics of each tumor. The myxoma was completely excised with a surgical margin, and the papillary fibroelastoma was shaved, preserving the cusp of the aortic valve. Pathohistologic examination confirmed the diagnosis. We discuss the echocardiographic characteristics and preferred surgical approaches for the two tumors. PMID- 19811575 TI - Open thoracic aortic repair for dysphagia in patients with Kommerell's diverticulum and right-sided aortic arch with aberrant left subclavian artery. AB - Kommerell's diverticulum of an aberrant left subclavian artery associated with a right-sided aortic arch is a rare congenital aortic anomaly. This communication reports two cases of this congenital anomaly leading to severe dysphagia. Both cases were successfully repaired with a staged open approach rather than an endovascular approach, which would not have addressed the presenting symptom of dysphagia. PMID- 19811576 TI - Distortion of the proximal circumflex artery during mitral valve repair. AB - A 74-year-old woman showed electrocardiographic signs of severe lateral ischemia with no hemodynamic consequence after mitral valve repair for severe mitral regurgitation. An angiogram showed interruption of the proximal circumflex artery. The patients then underwent an on-pump beating heart marginal branch revascularization. A new angiogram performed before discharge showed a widely patent graft. PMID- 19811577 TI - Two-stage surgical repair for truncus arteriosus with unilateral absence of the left proximal pulmonary artery: translocation of the left pulmonary artery to the right pulmonary artery. AB - A two-stage surgical repair of a one-month-old infant with truncus arteriosus with unilateral absence of a proximal pulmonary artery (PA) with a closed ipsilateral ductus arteriosus was successfully performed. In the first palliation, translocation of the discontinuous, closed, and undeveloped PA to the adjacent area of the other PA was useful for making a pulmonary arterial confluence at timing of the subsequent radical operation. PMID- 19811578 TI - Reversal of paraplegia following repair of an ascending aortic dissection. AB - We report the case of a patient who presented with sudden onset of paraplegia following an acute ascending aortic dissection extending to the arch, descending, and abdominal aorta. The paraplegia was fully reversed following an emergent replacement of the ascending aorta and hemi-arch repair. PMID- 19811579 TI - Expression of inducible nitric oxide synthase in conduits used in patients with diabetes mellitus undergoing coronary revascularization. AB - BACKGROUND: Expression of inducible nitric oxide synthase (iNOS) is a marker of vascular inflammation which can result in thrombosis and atherosclerosis. This study was undertaken to examine the difference in iNOS expression in the internal mammary artery (IMA) and saphenous veins (SVs) of patients with diabetes mellitus undergoing coronary artery bypass graft (CABG) surgery using both qualitative and quantitative methodology. METHODS: Segments of IMA and SV harvested in 100 diabetic patients with diabetes mellitus undergoing CABG surgery were fixed in formalin and immunostained to detect the presence of iNOS. Sections were graded using a qualitative score (0 = absence of iNOS expression to 3 = extensive expression of iNOS) and a quantitative computer-aided image analysis (area of staining/area of endothelium). Linear regression analyses were performed to assess the association of the degree of iNOS expression in both the IMA and SV with the type of diabetes control (insulin, oral, diet), and the serum levels of HbAlc, glucose, free fatty acids (ffa), C-reactive protein (CRP), and low-density liproprotein (LDL) at the time of conduit harvest. RESULTS: The degree of iNOS expression was significantly lower in the IMA compared to the SV by both qualitative (0.88 +/- 0.74 SD IMA vs. 1.38 +/- 0.68 SV; p < 0.0001) and quantitative (11.76 +/- 3.34% IMA vs. 17.10 +/- 2.54% SV; p = 0.01) methods. The Spearman rank correlation analysis showed a highly statistically significant association between the two methodologies (p < 0.0001). There was no correlation between iNOS expression in either the IMA or SV and the type of diabetes control, or levels of HA1c, glucose, ffa, and CRP. However, there was a significant (p = 0.04) correlation between LDL and iNOS expression in the SV graft, but not the IMA. CONCLUSIONS: iNOS expression is significantly decreased in the IMA compared to the SV in patients with diabetes mellitus undergoing CABG surgery. The degree of iNOS expression is unrelated to the level of glycemic control at the time of conduit harvest, but is associated with serum LDL levels in the SV, but not in the IMA grafts. PMID- 19811580 TI - The gingival biotype assessed by experienced and inexperienced clinicians. AB - AIM: A recent cluster analysis has identified three gingival biotypes among 100 periodontally healthy subjects based on different combinations of morphometric data related to maxillary front teeth and surrounding soft tissues. Patients with a thin-scalloped biotype are considered at risk because they have been associated with a compromised soft tissue response following surgical and/or restorative therapy. Hence, an accurate identification of these high-risk patients is warranted. The purpose of the present study was to evaluate the precision of simple visual inspection as a method to identify the gingival biotype by experienced and inexperienced clinicians. MATERIAL AND METHODS: Fifteen clinicians (five Restorative Dentists, five Periodontists and five Students) were invited to assess the gingival biotype (thin-scalloped, thick-flat, thick scalloped) of 100 periodontally healthy subjects based on clinical slides. Cluster analysis on these subjects was used as the gold standard and the accuracy in identifying the gingival biotype was determined using percentile agreement and kappa statistics. Intra- and inter-examiner reliability were also calculated. RESULTS: The gingival biotype was accurately identified only in about half of the cases irrespective of the clinician's experience. The thick-flat biotype was mostly recognized especially by experienced clinicians (> or =70% of the cases). Nearly half of the thin-scalloped cases were misclassified. The intra-examiner repeatability was fair to substantial (kappa: 0.328-0.670) and the inter-examiner reproducibility was slight to moderate (kappa: 0.127-0.547). CONCLUSIONS: Simple visual inspection may not be considered a valuable method to identify the gingival biotype as nearly half of the high-risk patients are overlooked. PMID- 19811581 TI - Characteristics of 2000 patients who visited a halitosis clinic. AB - AIMS: The aim of this paper was to analyse the aetiology and characteristics of 2000 patients who visited a multidisciplinary bad breath clinic in Leuven, Belgium and to correlate organoleptic ratings with portable device measurements. MATERIALS AND METHODS: The characteristics and aetiology of breath malodour of two thousand consecutive patients who visited a halitosis consultation were explored by means of a standard questionnaire and a clinical examination, including organoleptic scores provided by a trained and calibrated judge, and a portable bad breath detector (Halimeter). RESULTS: Most patients came without referral and had complaints for several years (mean: 7 years, SD: 8 years). For 76% of the patients, an oral cause was found [tongue coating (43%), gingivitis/periodontitis (11%) or a combination of the two (18%)]. Pseudo halitosis/halitophobia was diagnosed in 16% of the cases; and ear, nose and throat/extra-oral causes were found in 4% of the patients. Most patients had an organoleptic score <3 and a Halimeter value <240 p.p.b. CONCLUSIONS: Even though it was observed that halitosis has a predominantly oral origin, a multidisciplinary approach remains necessary to identify ear, nose and throat or extra-oral pathologies and/or pseudo-halitosis/halitophobia. PMID- 19811582 TI - Low prevalence of subgingival viruses in periodontitis patients. AB - BACKGROUND: Viruses such as Human Cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) have been proposed to be periodontal pathogens. The aim of this study was to analyse the presence of herpesvirus DNA in subgingival plaque samples of patients with different forms of periodontitis and in healthy periodontia. MATERIALS AND METHODS: A total of 140 ethnically mixed (prevalently Caucasian) subjects took part in the study. Sixteen were affected by localized aggressive periodontitis (LAgP), 64 by generalized aggressive periodontitis (GAgP), 20 by chronic periodontitis (CP) and 40 were periodontally healthy. Polymerase chain reaction (PCR) analyses were performed to detect HCMV and EBV. Sera were tested for anti-HCMV and EBV IgG antibodies. PCRs for herpes simplex (HSV) and varicella zoster virus (VZV) were performed in subgingival samples from a subset of 20 AgP subjects. RESULTS: HCMV DNA was not detected in any plaque samples. EBV DNA was detected in four LAgP (25%), two GAgP (3%) subjects and four healthy individuals (10%). HSV DNA and VZV DNA were not detected in the subset of studied individuals. CONCLUSIONS: This study challenges the previously reported high prevalence of herpesvirus DNA in subgingival samples from periodontitis patients and so questions whether they act as pathogens in such patients. PMID- 19811583 TI - Partial least squares path modelling for relations between baseline factors and treatment outcomes in periodontal regeneration. AB - BACKGROUND: Some clinical outcome variables in periodontal research are mathematically coupled, and it is not feasible to include all the mathematically coupled variables in an ordinary least squares (OLS) regression analysis. The simplest solution to this problem is to drop at least one of the mathematically coupled variables. However, this solution is not satisfactory when the mathematically coupled variables have distinctive clinical implications. MATERIAL AND METHODS: Partial least squares (PLS) methods were used to analyse data from a study on guided tissue regeneration. Relationships between characteristics of baseline lesions and treatment outcomes after 1 year were analysed using PLS, and the results were compared with those from OLS regression. RESULTS: PLS analysis suggested that there were multiple dimensions in the characteristics of baseline lesion: vertical dimension was positively associated with probing pocket depth (PPD) reduction and clinical attachment level (CAL) gain, whilst horizontal dimension was negatively associated with the outcome. Baseline gingival recession had a negative association with PPD reduction but a small positive one with CAL gain. CONCLUSION: PLS analysis provides new insights into the relationships between baseline characteristics of infrabony defects and periodontal treatment outcomes. The hypothesis of multiple dimensions in baseline lesions needs to be validated by further analysis of different datasets. PMID- 19811584 TI - The impact of powered and manual toothbrushing on incipient gingival recession. AB - AIM: To compare clinical effects of manual and powered toothbrushes on sites of localized gingival recession over 12 months. To evaluate patterns and the extent of toothbrush bristle wear. METHODS: A longitudinal, single-blind, randomized, parallel group clinical trial compared the effects of one manual and one powered toothbrush on incipient lesions of localized gingival recession. Toothbrush wear was evaluated concurrently by wear index and wear rating. RESULTS: Sixty patients were recruited and randomized to two groups with 52 (26 per group) attending the final visit at month 12. There were no differences between groups for full-mouth plaque index, pocket depth or bleeding on probing at baseline and month 12. There were no differences at target sites for clinical attachment level, pocket depth, bleeding on probing, plaque index, width of keratinized gingiva or maximal height of recession. There were no differences between the wear of the brushes as measured by wear index or wear rating. CONCLUSION: There was no progression of gingival recession in subjects using either toothbrush over 12 months. There was no difference in the overall wear of the powered and manual toothbrushes over successive 3-month periods. PMID- 19811585 TI - Correspondence between physiological and self-report measures of emotion dysregulation: a longitudinal investigation of youth with and without psychopathology. AB - BACKGROUND: Several theoretical perspectives suggest that emotion dysregulation is a predisposing risk factor for many psychiatric disorders. Yet despite a rapidly evolving literature, difficulties with emotion regulation (ER) are often measured inconsistently across studies, with little regard to whether different approaches capture the same construct. In this study, we evaluate the correspondence between two widely used measures of emotion dysregulation that cut across self-report and physiological levels of analysis. Our objectives were to (1) evaluate whether youth self-reports of ER difficulties correspond with physiological measures of emotion dysregulation collected at baseline and during sad emotion induction, and (2) validate the Difficulties in Emotion Regulation Scale (DERS) in a youth sample. METHOD: We measured emotion dysregulation among a sample of youth with depression, conduct problems, comorbid depression/conduct problems, or no psychiatric condition. Youth were assessed initially at ages 8-12 (Year 1) and followed up at Years 2 and 3. Respiratory sinus arrhythmia (RSA), a widely used physiological index of emotion regulation, was measured across all three years during sad emotion induction. At Year 3, the DERS was also administered. RESULTS: Multilevel modeling analyses indicated that slopes in RSA collected across the three assessments were associated with later self-reported ER abilities at the transition into adolescence. These findings were replicated across contexts (baseline and emotional challenge), suggesting that adolescents whose physiological responding to emotional challenge improves also experience fewer difficulties with emotion regulation as they mature. PMID- 19811586 TI - Chronic and acute stress, gender, and serotonin transporter gene-environment interactions predicting depression symptoms in youth. AB - BACKGROUND: Many recent studies of serotonin transporter gene by environment effects predicting depression have used stress assessments with undefined or poor psychometric methods, possibly contributing to wide variation in findings. The present study attempted to distinguish between effects of acute and chronic stress to predict depressive symptoms at age 20 among 346 youth varying in polymorphisms of the 5HTT gene who had been assessed at ages 15 and 20. METHODS: Interview measures assessed major acute life events between 15 and 19, and multiple interviews and questionnaires with youths and their parents at youth age 15 provided an index of chronic family stress. Lg alleles were reclassified as S. RESULTS: Chronic family stress at age 15 predicted higher depression scores at 20 among those with one or two S alleles, and the effects of genetic moderation were significant only for females. Gene-environment interactions with acute stress were nonsignificant. CONCLUSIONS: Careful measurement and separation of the effects of chronic and acute stress, and gender, are encouraged in the study of mechanisms of the stress-depression association. PMID- 19811587 TI - Primary cutaneous myxoid spindle cell squamous cell carcinoma: a clinicopathologic study and review of the literature. AB - Mucocutaneous squamous cell carcinoma (SCC) may rarely exhibit intracellular mucin production. Extracellular mucin production is an even rarer finding in SCC that is not well documented in the literature. Here, we report six cases of primary cutaneous and mucocutaneous SCC with prominent extracellular stromal mucin deposition and an epithelial spindle cell component. We propose the term 'yxoid spindle cell SCC' (MSC SCC) to describe the histologic characteristics of these six cases. We also propose a set of histologic and immunohistochemical findings for distinguishing MSC SCC from primary cutaneous and metastatic spindle cell neoplasms including other sarcomatoid carcinomas, myxoid sarcomas and the spindle cell variant of atypical fibroxanthoma (AFX). The criteria can also help discern MSC SCC from spindle cell melanomas, which may rarely show a prominent myxoid stroma. Given the small numbers of cases reported to date, the presence of prominent myxoid stroma in primary cutaneous spindle cell SCC has unknown prognostic significance at this time. PMID- 19811588 TI - Non-invasive risk stratification of coronary artery disease: an evaluation of some commonly used statistical classifiers in terms of predictive accuracy and clinical usefulness. AB - OBJECTIVE: To compare different prediction models for assessing outcome of patients undergoing non-invasive risk stratification of suspected or known coronary artery disease. METHODS: Six statistical classifiers and data mining models were applied to the prospective data bank of two different institutions. Of these, one represented the training (n = 2777) and the other one the test (n = 2679) set, each set consisting of usual clinical and stress echo information of patients followed-up for the combined endpoint of all-cause mortality and non fatal acute coronary syndromes. The following models were used: Logistic regression, Generalized Additive Model, Projection Pursuit Regression, Linear Discriminant Analysis, Quadratic Discriminant Analysis and Artificial Neural Networks. Models were selected using the Akaike Information Criterion and compared in terms of accuracy and Negative Predictive Value, overall Misclassification Rate and ROC Area Under Curve. RESULTS: During a median follow up of 31 months, 573 events occurred: 271 in the training and 302 in the test set respectively. All models selected the same subset of covariates as significantly associated with the outcome. The comparison of model performance showed that: (1) Quadratic Discriminant Analysis and Artificial Neural Networks provided a worse prediction of outcome than models more closely bonded to the hypothesis of linearity of the covariates effect; (2) overall predictive capability of the best performing models was excellent (>90% and >85% for training and test set respectively); and (3) there was a substantial lack of agreement among model indications in the individual patient. CONCLUSIONS: The selection of variables and predictive models are not independent processes and may affect the performance of risk scoring systems or algorithms designed to transfer general prognostic rules into clinical practice. Thus, caution must be used in translating model prediction into strict clinical indications. PMID- 19811589 TI - Do pathways lead to better organized care processes? AB - AIM: Clinical pathways are used worldwide to (re)organize care processes. They are used by multidisciplinary teams in their search towards excellence. The goal of this study is (1) to assess differences in the perception of health professionals in their evaluation of care processes; (2) to assess whether care processes supported by clinical pathways perform better than those not supported by clinical pathways; and (3) to assess the sensitivity and specificity of clinical pathways in predicting well-organized care processes. METHODS: A cross sectional, multi-centre study was performed comprising 309 healthcare workers, 103 care processes and 49 hospitals. The Care Process Self Evaluation Tool (CPSET) was used to score care processes according to their organization. Processes were also scored according to the level of pathway implementation. RESULTS: (1) Significant differences between healthcare professionals were found on two of five CPSET subscales. No significant differences were found among the overall CPSET scores. (2) Care processes supported by pathways had the highest CPSET scores. Nonetheless, continuous follow-up is necessary. (3) Clinical pathways have significant impact on the coordination of care (odds ratio: 8.92), follow-up (odds ratio: 6.65) and overall CPSET score (odds ratio: 4.26). Clinical pathways have a positive impact on the organization of care processes. Not all pathways have high CPSET scores, and care processes without pathways can also be well organized. Continuous evaluation is essential. This is the first study to analyse how healthcare teams perceive the organization of care processes with respect to clinical pathways. Our findings are important for other quality improvement methods. PMID- 19811590 TI - First-trimester Down syndrome screening in women younger than 35 years old and cost-effectiveness analysis in Taiwan population. AB - OBJECTIVES: Outcome of the first-trimester Down syndrome screening in younger population was less reported before. We present the outcome of this screening in Taiwanese women younger than 35 years old. We also test whether or not the first trimester Down syndrome screening of women <35 years of age and women >35 years old routinely receiving amniocentesis is cost-effective compared with all pregnant women screened with this test in the setting of increased maternal age. METHODS: From 1999 to 2007, the first-trimester Down syndrome screening including nuchal thickness, pregnancy-associated plasma protein A and free beta-hCG are provided to 10 811 singleton women <35 years of age with the cut-off of 1/270. A cost-effectiveness analysis of young women receiving this screening and older women undergo amniocentesis versus all women undergo this screening was performed in Taiwan population from 1987 to 2006, in which advanced age pregnancies increased from 2.8% to 11.6% of total pregnancies. RESULTS: Detection rates of trisomy 21, trisomy 18, Turner syndrome and other chromosome anormalies in women <35 years of age are 87.5% (14/16), 50% (2/4), 80% (8/10) and 63% (12/19), respectively, with a false-positive rate of 5.5% (590/10 811). As advanced age pregnancies reached 11.6%, the average cost per one case averted for all women screened ranged from $77 204 to $98 421, while the cost ranged from $99 647 to $116 433 for only women <35 years of age receiving this screening. CONCLUSIONS: In an aging population, the first-trimester Down syndrome screening should be implemented for all pregnant women when it is available. PMID- 19811591 TI - Computer-aided system for health economic evaluation. AB - BACKGROUND: Health policy makers are usually stranded by the complicated infrastructure and intensive computation related to economic evaluation. OBJECTIVES: It is therefore valuable to develop a computer-aided tool to help health personnel to perform economic evaluation with ease. METHOD: The infrastructure for economic evaluation was first designed. Markov process with micro-simulation was applied to model the disease natural history or lifetime sequale to project the effectiveness by comparing all possible decisions. All the essential elements of economic evaluation together with sensitivity analysis are encoded in this computer-aided software written with SAS Screen Control Language in user-defined menu style. ILLUSTRATION: Screening versus no screening for colorectal cancer was used as an example. CONCLUSION: The computer-aided model for economic evaluation was developed in this study. It is anticipated that the flexibility and user-defined menu style facilitate the wide application of economic evaluation to health care intervention program. PMID- 19811592 TI - Misleading one detail: a preventable mode of diagnostic error? AB - RATIONALE, AIMS AND OBJECTIVES: Despite advances in our understanding of cognitive biases in clinical practice, little is known about correction or prevention of diagnostic errors. The presence of a single misleading detail may lead clinicians down a cognitive and actual path toward an incorrect diagnosis. METHODS: In a large teaching hospital, we surveyed 51 attending doctors in internal medicine, presenting each with 10 clinical vignettes and soliciting their diagnosis of the condition leading to the presentation. Each of the 10 clinical cases included a single misleading detail. RESULTS: This survey elicited a wrong diagnosis in 90% of cases, which was reduced to 30% when omitting the misleading detail from the vignette. Diagnostic accuracy did not improve by warning doctors about potentially misleading information. Asking doctors to identify a leading diagnostic detail and then to formulate an alternative diagnosis after omission of the detail, significantly reduced diagnostic error rate by nearly 50%. CONCLUSION: Systematic re-examination of leading diagnostic clues may help to reduce errors in diagnosis. PMID- 19811593 TI - A RCT evaluating the effectiveness and cost-effectiveness of academic detailing versus postal prescribing feedback in changing GP antibiotic prescribing. AB - PURPOSE: The aim of this study is to evaluate the effectiveness of academic detailing (AD) plus postal prescribing feedback versus postal prescribing feedback alone in reducing: (i) the overall rate of antibiotic; and (ii) proportion of second-line antibiotic prescribing. In addition, the cost effectiveness of an outreach prescriber adviser service versus a postal prescribing feedback service was evaluated. METHODS: Volunteer general practitioner practices (n = 98) were randomized to receive prescribing feedback via postal bulletin (PB) (n = 50) or academic detailing plus postal bulletin (AD) (n = 48). Data analysis was based on the HSE-primary care reimbursement service (HSE-PCRS) prescribing database. Regression (beta) coefficients, indicating proportion change in prescribing per month, and 95% confidence intervals (CIs) are presented. The cost-effectiveness ratio was calculated from the total cost of the intervention divided by percentage change in antibiotic prescribing in AD versus PB group. RESULTS: Immediately post intervention PB (beta = -0.02, 95% CI 0.04, -0.001) and AD (beta = -0.02, 95% CI -0.03, -0.001) practices significantly decreased overall antibiotic prescribing. Second-line antibiotic prescribing was also significantly decreased by 2-3% in both groups. However, there were no significant differences in antibiotic prescribing between the randomized groups in the immediate or long-term post-intervention period. In the cost-effectiveness analysis a postal prescribing feedback service would cost euro 88 per percentage change in prescribing practice compared with euro 778 for a prescriber adviser service. CONCLUSION: Prescribing feedback significantly reduced overall and second-line antibiotic prescribing, but academic detailing was not significantly more effective than postal bulletin in changing antibiotic prescribing practice. PMID- 19811594 TI - Effectiveness of internal quality assurance programmes in improving clinical practice and reducing costs. AB - RATIONALE, AIMS AND OBJECTIVES: To assess the effectiveness of internal initiatives to improve quality as compared with external feedback, and as compared with a control group. METHOD: Ten primary health centres were randomly selected from the centres in the Murcia's Region and were randomly assigned into three groups: G1 committed themselves to the improvement; G2 composed by two subgroups: with and without quality improvement (QI) activities - received external feedback; G3 received no intervention. Quality of common cold management was measured in a random sample of 50 patients per centre before and after interventions. Effect was assessed comparing criteria compliance and the cost of treatments for common cold in the three groups. RESULTS: G1 and G2 subgroup with internal QI improved significantly in all criteria, and in G1 average treatment cost decreased by 60% (P < 0.01). G3 improved only in one criterion. Estimated yearly savings in treatment costs for common cold, for a centre with internal QI, were euro 12,193.83 and it would be euro 1,817 004.65 for the 30 centres in the Region. CONCLUSIONS: Internally assumed QI activities were significantly more effective than external feedback. Besides, the high cost of deficient quality provides a wide margin to invest with benefits in the promotion of internal QI programmes. PMID- 19811596 TI - The utility of the Illness Perception Questionnaire in the evaluation of mental health practitioners' perspectives on patients with schizophrenia. AB - RATIONALE, AIMS, OBJECTIVES: The Illness Perception Questionnaire (IPQ) was designed as a measure of illness perception and has been used extensively for measuring the perceptions of people with physical illness. Latterly modified versions of the IPQ have been used to measure the illness perceptions of people with mental health problems and their carers. This study examined the utility of a modified version of the IPQ to measure changes in mental health practitioners' illness perceptions about schizophrenia after undertaking psychosocial intervention training. METHOD: A total of 245 mental health practitioners who undertook psychosocial intervention training completed a modified version of the IPQ before and after training. The structure of the questionnaire was tested using confirmatory factor analysis. The internal consistency of sub-scales embedded in the instrument was also examined. RESULTS: The hypothesized structure failed to account for the data. Model fit indices revealed a poor fit to the data across all models evaluated. Cronbach's alpha revealed a number of sub-scales in the instrument to have mediocre internal consistency characteristics. CONCLUSIONS: The modified version of the IPQ is not suitable for evaluating the impact of psychosocial intervention training on changes in illness perceptions of schizophrenia in mental health practitioners. However, the study has highlighted the need to develop a valid and reliable measure to assess the illness perceptions health professionals have of patients in their care. PMID- 19811597 TI - A combined diabetes renal clinic improves risk factor management and progression of renal disease in a district general hospital. AB - AIMS: Aggressive management of vascular risk factors reduces the rate of progression to end-stage renal failure in patients with diabetic nephropathy. The aim of our audit was to clarify whether improvements in patient care could be demonstrated within a short time period after establishing a combined diabetes renal clinic in a district general hospital. METHODS: A retrospective analysis of weight, glycaemic control, blood pressure control, rate of decline in renal function and appropriate use of medications for vascular risk management for patients in a district general hospital was performed before and after attendance at a combined diabetes renal clinic. RESULTS: Data were complete for 45 patients. There was no significant change in weight from 14 months before attending the combined clinic, referral to the clinic and after 14 months of mean follow-up. An improvement in mean systolic blood pressure was achieved at the combined diabetes renal clinic (from 147 to 134 mmHg with an average fall of 16.5 mmHg, P < 0.01). The mean glycated haemoglobin (HbA1c) improved from 8.6% to 8% (P = 0.002). The rate of decline in estimated glomerular filtration rate of 1.16 mL min(-1) per month prior to referral improved to 0.21 mL min(-1) per month on attending the combined clinic (P = 0.002). CONCLUSIONS: Vascular risk factor management was improved and decline in renal function slowed in patients with diabetic nephropathy within a short period of establishing a combined diabetes renal clinic in a district general hospital. PMID- 19811598 TI - Do pre-printed clerking templates improve environmental history taking in the medical assessment unit? PMID- 19811595 TI - Responsiveness of the EQ-5D to HADS-identified anxiety and depression. AB - RATIONALE: The use of generic measures of health-related quality of life enables cost effectiveness comparisons of different health care interventions to be made. Nevertheless, there exists a concern that generic instruments may be insufficiently sensitive to detect the differences or changes in outcome identified by condition-specific instruments. This paper compares the psychometric properties of the EQ-5D generic instrument with a widely used specific measure of distress, the Hospital Anxiety and Depression Scale (HADS). METHOD: The analysis was based on data obtained from a large sample of women (n = 3119) with low-grade cervical cytological abnormalities detected at routine screening. These women completed EQ-5D and HADS questionnaires at recruitment and at 12 months thereafter. We examined the strength of association between HADS determined severity of distress and EQ-5D scores at recruitment and between changes in severity and in scores over time. RESULTS: A higher likelihood of HADS identified anxiety and/or depression was associated with significantly lower EQ 5D index and visual analogue scores. Over time, the EQ-5D score rose significantly when the likelihood of an individual representing a HADS-defined anxiety and/or depression case decreased. CONCLUSION: We conclude that the EQ-5D has shown itself to be responsive to differing degrees of HADS-assessed distress, although generalization beyond the UK context requires further investigation. PMID- 19811599 TI - Philosophy, freedom and the public good: a review and analysis of 'Public health ethics' Holland, S. (2007): Polity Press, Cambridge. ISBN-13: 978-07456-3302-2, pp. 222. PMID- 19811600 TI - Understanding and changing health systems--an instinctive and natural process? PMID- 19811601 TI - Part 1--unravelling primary health care conceptual predicaments through the lenses of complexity and political economy: a position paper for progressive transformation. AB - OBJECTIVE: To disentangle the concepts of primary health care and primary care as well as their conceptual and empirical ramifications for progressive transformation. METHODS: over 400 international and interdisciplinary abstracts and papers with 96 annotated bibliography abstracts of literature across multiple dimensions relating to the knowledge base around mechanisms in PHC development were reviewed. The text is confronted with the reality, as it exists in the field and makes the case for complexity perspectives to assess this phenomenon in its context. CONCLUSION: PHC complexity is an important analytical tool to interrogate the ways in which this phenomenon is socially constructed as well as in the matrices in which it is embedded. It is also a potent analytical tool to assist in the deconstruction of prevalent linear thinking built around PHC as a whole. PMID- 19811602 TI - Health promotion as a systems science and practice. AB - RATIONALE: Health promotion is where clinical practice and prevention science intersect to address complex or 'wicked' problems that have multiple sources and require a broad perspective to address. This means focusing on the social determinants of health and the complex individual, community and environmental interactions that influence health and wellbeing. Health promotion research and practice recognizes that social change is not linear and involves multiple communities of interest working together in a coordinated manner in order to address health problems. An approach that acknowledges this non-linear system of interaction in its data gathering, strategic planning, and program implementation is necessary to addressing this complexity in practice. METHODS: Concepts such as chaos theory, self-organization, social emergence can inform how health promotion is practiced at multiple levels. Evaluation approaches such as social network analysis, system dynamics modeling combined with social organizing strategies like communities of practice and unconferences provide opportunities to leverage social capital effectively to promote health in complex environments with diverse populations. CONCLUSION: Health promotion's focus on the multi-layered, complex interactions that create or limit health and wellbeing require knowledge and action that match this complexity. Approaches to engagement and evaluation that are based on systems theories and methodologies provide the means of addressing this complexity, while framing health promotion as a systems science and practice. PMID- 19811603 TI - Revitalizing primary health care and family medicine/primary care in India- disruptive innovation? AB - CONTEXT: India has rudimentary and fragmented primary health care (PHC) and family medicine systems, yet it also has the policy expectation that PHC should meet the needs of extremely large populations with slums and difficult to reach groups, rapid social and epidemiological transition from developing to developed nation profiles. Historically, the system has lacked impetus to achieve PHC. OBJECTIVE: To provide an overview of PHC approaches and the current state of PHC and family medicine in India in order to assess the opportunities for their revitalization. METHODS: A narrative review of the published and grey literature on PHC, family medicine, Web2.0 and health informatics key papers and policy documents, pertinent to India. OUTCOMES: A conceptual framework and recommendations for policy makers and practitioner audiences. FINDINGS: PHC is constructed through systems of local providers who address individual, family and local community basic health needs with strong community participation. Successful PHC is a pre-eminent strategy for India to address the determinants of health and the almost chaotic of massive social transition in its institutions and health care sector. There is a lack of an articulated comprehensive framework for the publicly stated goals of improving health and implementing PHC. Also, there exists a very limited education and organization of a medical and PHC workforce who are trained and resourced to address individual, family and local community health and who have become increasingly specialized. However, emerging technology, Health2.0 and user generated health care informatics, which are largely conducted through mobile phones, are co-evolving patient-driven health systems, and potentially enhance PHC and family medicine workforce development. CONCLUSIONS: In order to improve health outcomes in an equitable manner in India, there is a pressing need for a framework for implementing PHC. The co-emergence of information technologies accessible to the mass population and user-driven health care provide a potential catalyst or innovation for this transition. PMID- 19811604 TI - Time and the consultation--an argument for a 'certain slowness'. AB - When natural time sequences were replaced by clocks, time became a measurable commodity and the 'speedy use of time' a virtue. In medical practice shorter consultations allow more patients to be seen, whereas longer consultations result in a better understanding of the patient and her problems. Crossing the line of time-efficiency and time-effectiveness compromises the balance between short-term turnover and long-term outcomes. The consultation has all the hallmarks of a complex adaptive system whose characteristics are not determined by the characteristics of the components, but by the patterns of interaction among the components. Systems are dynamic and change over time; the dynamic nature is not incidental, but necessary as complex systems operate at conditions far from equilibrium. The central notion when we talk of time and complexity is that of 'memory'. Memory is carrying something from the past over into the future. Memory is filtered/interpreted, separating noise from information. Memory therefore is not an instantaneous thing, it takes time to develop; it is slow. The dynamics between the participating agents in the consultation will create shared memories that live on to shape future interactions. Shared memories are stronger and contain more relevant knowledge if they are based on frequent interactions and ongoing doctor-patient relationships, leading to a better understanding of the whole person - a process that takes time. Sufficient time, that is, 'a certain slowness', is an essential element of the healing relationship in the consultation. It creates a sufficiently stable, but adaptive, environment that can withstand changing demands. Hence a more complete understanding of the consultation and its time demands will not only lead to more effective treatment, it will also humanize a situation which has become to a large extent purely instrumental. This process of humanization is important not only for the patient, but also for the doctor. PMID- 19811605 TI - Analyses of amino acid sequences in hypervariable region-1 of hepatitis C virus (HCV) in sera from chimpanzees infected three times with the same HCV strain. AB - BACKGROUND: To investigate whether or not the same strain of hepatitis C virus (HCV) can twice re-infect the same chimpanzee, we analyzed nucleic and amino acid sequences in HCV hypervariable region-1 (HVR1). Two chimpanzees were inoculated, three times each, with the same HCV strain during the 1983-1991. After each inoculation, chimpanzees developed acute hepatitis C, and then recovered. METHODS: Using sera, HVR1 cloning and antibody to HVR1 major clone measurement were performed. RESULTS: Clones from the first inoculum were divisible into major and minor types. Clones from the second and third inocula, as well as all post inoculation sera, were essentially identical to the major type. Titers of antibody to HVR1 major clone were consistently low in pre- and post-inoculation sera. CONCLUSIONS: Both chimpanzees were re-infected twice with the same strain of HCV. The sequences from the second and third infections were similar to the major sequences in the first inoculum. PMID- 19811606 TI - Amyloid precursor protein 695 associates with assembled NR2A- and NR2B-containing NMDA receptors to result in the enhancement of their cell surface delivery. AB - This is a study of the interaction between the two NMDA neurotransmitter receptor subtypes, NR1/NR2A and NR1/NR2B, and amyloid precursor protein (APP) 695, the major APP variant expressed in neurones. APP695 co-immunoprecipitated with assembled NR1-1a/NR2A and NR1-1a/NR2B NMDA receptors following expression in mammalian cells. Single NR1-1a, NR1-2a, NR1-4b(c-Myc), or NR2 subunit transfections revealed that co-association of APP695 with assembled NMDA receptors was mediated via the NR1 subunit; it was independent of the NR1 C1, C2, and C2' cassettes and, the use of an NR1-2a(c-Myc)-trafficking mutant suggested that interaction between the two proteins occurs in the endoplasmic reticulum. The use of antibodies directed against extracellular and intracellular NR2 subunit epitopes for immunoprecipitations suggested that APP/NMDA receptor association was mediated via N-terminal domains. Anti-APP antibodies immunoprecipitated NR1, NR2A, and NR2B immunoreactive bands from detergent extracts of mammalian brain; reciprocally, anti-NR1 or anti-NR2A antibodies co immunoprecipitated APP immunoreactivity. Immune pellets from brain were sensitive to endoglycosidase H suggesting that, as for heterologous expression, APP and NMDA receptor association occurs in the endoplasmic reticulum. Co-expression of APP695 in mammalian cells resulted in enhanced cell surface expression of both NR1-1a/NR2A and NR1-1a/NR2B NMDA receptors with no increase in total subunit expression. These findings are further evidence for a role of APP in intracellular trafficking mechanisms. Further, they provide a link between two major brain proteins that have both been implicated in Alzheimer's disease. PMID- 19811607 TI - Functional effects of somatostatin receptor 1 activation on synaptic transmission in the mouse hippocampus. AB - Somatostatin-14 (SRIF) co-localizes with GABA in the hippocampus and regulates neuronal excitability. A role of SRIF in the control of hippocampal activity has been proposed, although the exact contribution of each SRIF receptor (sst(1) sst(5)) in mediating SRIF action requires some clarification. We used hippocampal slices of wild-type and sst(1) knockout (KO) mice and selective pharmacological tools to provide conclusive evidence for a role of sst(1) in mediating SRIF inhibition of synaptic transmission. With single- and double-label immunohistochemistry, we determined the distribution of sst(1) in hippocampal slices and we quantified sst(1) colocalization with SRIF. With electrophysiology, we found that sst(1) activation with CH-275 inhibited both the NMDA- and the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-mediated responses. Results from sst(1) KO slices confirmed the specificity of CH-275 effects; sst(1) activation did not affect the inhibitory transmission which was in contrast increased by sst(4) activation with L-803,087 in both wild-type and sst(1) KO slices. The AMPA-mediated responses were increased by L-803,087. Functional interaction between sst(1) and sst(4) is suggested by the finding that their combined activation prevented the CH-275-induced inhibition of AMPA transmission. The involvement of pre-synaptic mechanisms in mediating inhibitory effects of sst(1) on excitatory transmission was demonstrated by the finding that CH-275 (i) increased the paired-pulse facilitation ratio, (ii) did not influence the AMPA depolarization in the presence of tetrodotoxin, and (iii) inhibited glutamate release induced by epileptiform treatment. We conclude that SRIF control of excitatory transmission through an action at sst(1) may represent an important contribution to the regulation of hippocampal activity. PMID- 19811608 TI - Central amylin expression and its induction in rat dams. AB - Amylin, or islet amyloid polypeptide, is known as a satiating signal expressed in pancreatic beta-cells but not in the brain. In this study, regulations of postpartum mRNA expressions were investigated in the preoptic area of the hypothalamus. mRNA levels of lactating dams and mothers whose pups were removed immediately after delivery were compared in microarray experiments. There were 20 genes identified with significantly increased and 14 with decreased expression 9 days postpartum. Amylin mRNA level demonstrated the largest change, a 25.7 times increase. Quantitative RT-PCR measurements validated the increase in the mRNA level of amylin in the preoptic area of lactating dams while the expression level of other members of the calcitonin gene-related peptide family did not change. In situ hybridization histochemistry for amylin further verified its induction in lactating mothers and demonstrated the distribution of amylin mRNA in the medial preoptic nucleus, parts of the medial preoptic area, and the ventral part of the bed nucleus of the stria terminalis but nowhere else in the rat brain. Immunolabeling verified the postpartum induction of amylin in the preoptic area at the peptide level, as well. The results suggest that amylin may play a part in maternal regulations. PMID- 19811609 TI - A multicenter analysis of "time to microcatheter" for endovascular therapy in acute ischemic stroke. AB - PURPOSE: At present, no time recommendation for initiation of endovascular treatment in acute ischemic stroke is available. A multicenter analysis was designed to identify variables that prolong "time to microcatheter," defined as the time interval from computed tomographic scan to microcatheter placement in the cerebral circulation. METHODS: Consecutive acute ischemic stroke patients from 3 academic stroke centers were included. Analysis of covariance was used to evaluate different variables that prolong "time to microcatheter." RESULTS: Ninety-one patients underwent emergent endovascular treatment for acute ischemic stroke. Mean "time to microcatheter" was 174+/-60 minutes. No significant time difference was found in patients who were intubated, presented at night or weekends, were administered intravenous recombinant tissue plasminogen activator, or underwent additional imaging prior to endovascular treatment. "Time to microcatheter" was significantly longer in nonlevel I trauma centers and in patients with National Institutes of Health Stroke Scale Score of 10 to 19. CONCLUSION: Wide variability of "time to microcatheter" among institutions highlights the need for standardized time goals. PMID- 19811610 TI - Ongoing transmission of a single hepatitis B virus strain among men having sex with men in Amsterdam. AB - For the past decade, a specific hepatitis B virus (HBV) genotype A strain has been prevalent among men having sex with men (MSM) in Amsterdam, the Netherlands. At what point in time this strain was introduced in the MSM population, and why only this specific strain continues to be transmitted, remains unclear. Between 1984 and 2003, sera of 1862 MSM were retrospectively screened for anti-HBc in the context of the Amsterdam Cohort studies. After 2003, most MSM participating in this study were vaccinated, making further testing less useful. HBV DNA from anti HBc seroconverters was amplified and sequenced. Poisson regression was used to test for temporal trends in HBV and HIV incidence. Of the 1042 MSM who were negative for anti-HBc at entry, 64 had seroconverted during follow-up at a median age of 32. At the point of seroconversion, 31 MSM were HIV positive. HBV incidence declined dramatically in the first years and then remained stable throughout the study period. The HBV and HIV incidence ran almost in parallel. With the exception of three MSM, all were infected with genotype A. Fifteen of these (41%) were infected with an identical genotype A strain. For the past two decades, an identical genotype A strain has been circulating among MSM in the Netherlands. Although HBV is generally considered more infectious than HIV, this study shows that the trend and magnitude in HBV and HIV incidence among MSM are similar. PMID- 19811611 TI - Autonomic dysfunction: often present but usually ignored in patients with liver disease. PMID- 19811612 TI - Pretransplantation calcineurin activity: towards individualized therapy? PMID- 19811613 TI - Pharmacological actions of curcumin in liver diseases or damage. AB - Since 1900 bc, several therapeutic activities have been attributed to the rhizomes of the plant Curcuma longa for a variety of diseases, including liver disorders. Curcumin, the main active compound obtained from this plant, was first isolated two centuries ago and its structure as diferuloylmethane was determined in 1910. Curcumin has shown anti-inflammatory, anti-oxidant, antifungal, antibacterial and anticancer activities. The pharmacological properties of curcumin were reviewed recently and focused mainly on its anticancer properties. However, its beneficial activity on liver diseases (known centuries ago, and demonstrated recently utilizing animal models) has not being reviewed in depth until now. The curcumin ability to inhibit several factors like nuclear factor kappaB, which modulates several pro-inflammatory and profibrotic cytokines as well as its anti-oxidant properties, provide a rational molecular basis to use it in hepatic disorders. Curcumin attenuates liver injury induced by ethanol, thioacetamide, iron overdose, cholestasis and acute, subchronic and chronic carbon tetrachloride (CCl(4)) intoxication; moreover, it reverses CCl(4) cirrhosis to some extent. Unfortunately, the number of studies of curcumin on liver diseases is still very low and investigations in this area must be encouraged because hepatic disorders constitute one of the main causes of worldwide mortality. PMID- 19811614 TI - Subcutaneous phaeohyphomycosis in immunocompetent patients: two new cases caused by Exophiala jeanselmei and Cladophialophora carrionii. AB - Phaeohyphomycosis is a distinct mycotic infection of the skin or internal organs caused by darkly pigmented (dematiaceous) fungi, which are widely distributed in the environment. Phaeohyphomycosis is most frequently an opportunistic infection in immunosuppressed patients (HIV, corticotherapy, transplant patients) or is frequently associated with chronic diseases and diabetes. The spectrum of the disease is broad and includes superficial infections, onychomycosis, subcutaneous infections, keratitis, allergic disease, pneumonia, brain abscesses and disseminated disease. Rarely, immunocompetent patients may be affected. We describe two new cases of subcutaneous phaeohyphomycosis in immunocompetent patients: in the first patient, the causative agent was Exophiala jeanselmei, a common cause of phaeohyphomycosis; and in the second, Cladophialophora carrionii, which could be identified by culture. Cladophialophora carrionii is mainly the aetiological agent of chromoblastomycosis and only rarely the cause of phaeohyphomycosis. The first patient was treated with surgical excision and oral itraconazole, and the second patient responded to oral itraconazole only. Lesions improved in both patients and no recurrence was observed at follow-up visits. PMID- 19811615 TI - Therapeutic efficacy of posaconazole against Candida glabrata in a murine model of vaginitis. AB - The frequency of mucosal infections caused by Candida glabrata has increased significantly. Candida glabrata infections are often resistant to many azole antifungal agents, especially fluconazole. The purpose of this study was to compare the efficacies of posaconazole (PSC) and fluconazole (FLC) in the treatment of experimental C. glabrata vaginitis caused by isolates with different FLC susceptibilities. A battery of 36 vaginal isolates of C. glabrata was tested against PSC and FLC to determine their in vitro susceptibilities. The 48-h geometric mean MICs for all isolates tested were 0.156 and 4.238 MUg ml(-1) for PSC and FLC respectively. Two strains of C. glabrata for which FLC MICs were different were selected for in vivo study. The treatment regimens for the vaginal murine infection model were PSC or FLC at 10 or 20 mg kg(-1) of body weight/day and 20 mg kg(-1) twice a day. Regimens with PSC at 20 mg kg(-1) once or twice a day were effective in reducing the load of both the FLC-susceptible and resistant isolates of C. glabrata. FLC at 20 mg kg(-1) twice a day was effective in reducing the load of both the isolates of C. glabrata. PSC displayed a more effective in vivo activity than FLC in the treatment of murine C. glabrata vaginitis. PMID- 19811616 TI - Nuclear localization of Ca(v)2.2 and its distribution in the mouse central nervous system, and changes in the hippocampus during and after pilocarpine induced status epilepticus. AB - AIMS: To investigate the subcellular localization of Ca(v)2.2 calcium channel in the mouse central nervous system (CNS), and changes of Ca(v)2.2 at acute and chronic stages during and after pilocarpine-induced status epilepticus (PISE), in order to find out the roles it may play in epileptogenesis. METHODS: Combined immunocytochemistry at both light and electron microscopic levels with real-time reverse transcription polymerase chain reaction (RT-PCR), cell transfection approach were used in this study. RESULTS: N-type calcium channel Ca(v)2.2 subunit was distributed in different regions of the mouse CNS. It was mainly localized in the nuclei in different types of neurones and in astrocytes. At acute stages during and after PISE, Ca(v)2.2 expression decreased in the stratum pyramidale of CA3 area and in the stratum granulosum of the dentate gyrus, but increased in the stratum lucidum of CA3 area and in the hilus of the dentate gyrus. At chronic stage at 2 months after PISE, increased expression of Ca(v)2.2 in both the strata granulosum and molecular of the dentate gyrus was observed. CONCLUSIONS: Ca(v)2.2 is a nuclear protein in neurones and astrocytes in the mouse CNS. Its translocation occurs at acute stages during and after PISE. The increased expression of Ca(v)2.2 in both the strata granulosum and moleculare of the dentate gyrus at chronic stage at 2 months after PISE may be involved in the occurrence of spontaneously recurrent seizures. PMID- 19811617 TI - Silent subtype 3 carcinoma of the pituitary: a case report. PMID- 19811619 TI - Probing differentially expressed genes against a microarray database for in silico suppressor/enhancer and inhibitor/activator screens. AB - High-density oligonucleotide arrays are widely used for analysis of gene expression on a genomic scale, but the generated data remain largely inaccessible for comparative analysis purposes. Similarity searches in databases with differentially expressed gene (DEG) lists may be used to assign potential functions to new genes and to identify potential chemical inhibitors/activators and genetic suppressors/enhancers. Although this is a very promising concept, it requires the compatibility and validity of the DEG lists to be significantly improved. Using Arabidopsis and human datasets, we have developed guidelines for the performance of similarity searches against databases that collect microarray data. We found that, in comparison with many other methods, a rank-product analysis achieves a higher degree of inter- and intra-laboratory consistency of DEG lists, and is advantageous for assessing similarities and differences between them. To support this concept, we developed a tool called MASTA (microarray overlap search tool and analysis), and re-analyzed over 600 Arabidopsis microarray expression datasets. This revealed that large-scale searches produce reliable intersections between DEG lists that prove to be useful for genetic analysis, thus aiding in the characterization of cellular and molecular mechanisms. We show that this approach can be used to discover unexpected connections and to illuminate unanticipated interactions between individual genes. PMID- 19811618 TI - Reverse breeding: a novel breeding approach based on engineered meiosis. AB - Reverse breeding (RB) is a novel plant breeding technique designed to directly produce parental lines for any heterozygous plant, one of the most sought after goals in plant breeding. RB generates perfectly complementing homozygous parental lines through engineered meiosis. The method is based on reducing genetic recombination in the selected heterozygote by eliminating meiotic crossing over. Male or female spores obtained from such plants contain combinations of non recombinant parental chromosomes which can be cultured in vitro to generate homozygous doubled haploid plants (DHs). From these DHs, complementary parents can be selected and used to reconstitute the heterozygote in perpetuity. Since the fixation of unknown heterozygous genotypes is impossible in traditional plant breeding, RB could fundamentally change future plant breeding. In this review, we discuss various other applications of RB, including breeding per chromosome. PMID- 19811620 TI - A petunia chorismate mutase specialized for the production of floral volatiles. AB - In Petunia x hybrida cv. 'Mitchell Diploid' floral fragrance is comprised of 13 volatile benzenoids/phenylpropanoids derived from the aromatic amino acid phenylalanine. Several genes involved in the direct synthesis of individual floral volatile benzenoid/phenylpropanoid (FVBP) compounds, i.e. at the end of the pathway, have been isolated and characterized in petunia through reverse genetic and biochemical approaches. In an effort to understand the regulation of 'upstream' components in the FVBP system, we have cloned and characterized two CHORISMATE MUTASE (PhCM1 and PhCM2) cDNAs from petunia. PhCM1 has a transcript accumulation profile consistent with known FVBP genes, while PhCM2 showed a constitutive transcript accumulation profile. The plastid-localized PhCM1 is allosterically regulated by tryptophan but not phenylalanine or tyrosine. The total FVBP emission in PhCM1 RNAi knockdown petunias is reduced by approximately 60-70%, and total chorismate mutase activity in corolla tissue is reduced by 80 85% compared to control plants. These results show that PhCM1 is the principal CHORISMATE MUTASE responsible for the coupling of metabolites from the shikimate pathway to the synthesis of FVBPs in the corolla of Petunia x hybrida cv. 'Mitchell Diploid'. PMID- 19811621 TI - Heritable targeted mutagenesis in maize using a designed endonuclease. AB - The liguleless locus (liguleless1) was chosen for demonstration of targeted mutagenesis in maize using an engineered endonuclease derived from the I-CreI homing endonuclease. A single-chain endonuclease, comprising a pair of I-CreI monomers fused into a single polypeptide, was designed to recognize a target sequence adjacent to the LIGULELESS1 (LG1) gene promoter. The endonuclease gene was delivered to maize cells by Agrobacterium-mediated transformation of immature embryos, and transgenic T(0) plants were screened for mutations introduced at the liguleless1 locus. We found mutations at the target locus in 3% of the T(0) plants, each of which was regenerated from independently selected callus. Plants that were monoallelic, biallelic and chimeric for mutations at the liguleless1 locus were found. Relatively short deletions (shortest 2 bp, longest 220 bp) were most frequently identified at the expected cut site, although short insertions were also detected at this site. We show that rational re-design of an endonuclease can produce a functional enzyme capable of introducing double-strand breaks at selected chromosomal loci. In combination with DNA repair mechanisms, the system produces targeted mutations with sufficient frequency that dedicated selection for such mutations is not required. Re-designed homing endonucleases are a useful molecular tool for introducing targeted mutations in a living organism, specifically a maize plant. PMID- 19811622 TI - Blame apportioning and the emergence of zoonoses over the last 25 years. AB - Over the last 25 years zoonoses have emerged and re-emerged worldwide without a specific pattern or trend. The most recent ones (1998-2008) are geographically limited to Southeast Asia, making this salient region the most immediate culprit. The media, experts and politicians have helped shape public perceptions regarding the origin of emerging zoonoses. Assigning blame to certain countries or regions can results in social, political, cultural and economic consequences that turn out to be unjustified, unfair and ultimately detrimental. PMID- 19811623 TI - The analysis of crow population dynamics as a surveillance tool. AB - West Nile virus (WNV) infection, a zoonotic disease for which birds act as a reservoir, first appeared in North America in August 1999. It was first reported in Quebec in 2002. The Quebec surveillance system for WNV has several components, including the surveillance of mortality in corvid populations, which includes the American crow (Corvus brachyrhynchos). The main objectives of this study are to better understand the population dynamics of this species in Quebec and to evaluate the impact of WNV on these dynamics. We obtained observation data for living crows in this province for the period of 1990-2005 and then conducted a spectral analysis of these data. To study changes in crow population dynamics, the analysis was carried out before and after the appearance of WNV and space was divided in two different areas (urban and non-urban). Our results show the importance of cycles with periods of less than 1 year in non-urban areas and cycles with periods of greater than 1 year in urban areas in the normal population dynamics of the species. We obtained expected fluctuations in bird densities using an algorithm derived from spectral decomposition. When we compared these predictions with data observed after 2002, we found marked perturbations in population dynamics beginning in 2003 and lasting up to 2005. In the discussion, we present various hypotheses based on the behaviour of the American crow to explain the normal population dynamics observed in this species and the effect of type of area (urban versus non-urban). We also discuss how the predictive algorithm could be used as a disease surveillance tool and as a measure of the impact of a disease on wild fauna. PMID- 19811624 TI - Interdisciplinary diabetes care teams operating on the interface between primary and specialty care are associated with improved outcomes of care: findings from the Leuven Diabetes Project. AB - BACKGROUND: Type 2 diabetes mellitus is a complex, progressive disease which requires a variety of quality improvement strategies. Limited information is available on the feasibility and effectiveness of interdisciplinary diabetes care teams (IDCT) operating on the interface between primary and specialty care. A first study hypothesis was that the implementation of an IDCT is feasible in a health care setting with limited tradition in shared care. A second hypothesis was that patients who make use of an IDCT would have significantly better outcomes compared to non-users of the IDCT after an 18-month intervention period. A third hypothesis was that patients who used the IDCT in an Advanced quality Improvement Program (AQIP) would have significantly better outcomes compared to users of a Usual Quality Improvement Program (UQIP). METHODS: This investigation comprised a two-arm cluster randomized trial conducted in a primary care setting in Belgium. Primary care physicians (PCPs, n = 120) and their patients with type 2 diabetes mellitus (n = 2495) were included and subjects were randomly assigned to the intervention arms. The IDCT acted as a cornerstone to both the intervention arms, but the number, type and intensity of IDCT related interventions varied depending upon the intervention arm. RESULTS: Final registration included 67 PCPs and 1577 patients in the AQIP and 53 PCPs and 918 patients in the UQIP. 84% of the PCPs made use of the IDCT. The expected participation rate in patients (30%) was not attained, with 12,5% of the patients using the IDCT. When comparing users and non-users of the IDCT (irrespective of the intervention arm) and after 18 months of intervention the use of the IDCT was significantly associated with improvements in HbA1c, LDL-cholesterol, an increase in statins and anti-platelet therapy as well as the number of targets that were reached. When comparing users of the IDCT in the two intervention arms no significant differences were noted, except for anti-platelet therapy. CONCLUSION: IDCT's operating on the interface between primary and specialty care are associated with improved outcomes of care. More research is required on what team and program characteristics contribute to improvements in diabetes care. TRIAL REGISTRATION: NTR 1369. PMID- 19811625 TI - Estimation of physician supply by specialty and the distribution impact of increasing female physicians in Japan. AB - BACKGROUND: Japan has experienced two large changes which affect the supply and distribution of physicians. They are increases in medical school enrollment capacity and in the proportion of female physicians. The purpose of this study is to estimate the future supply of physicians by specialty and to predict the associated impact of increased female physicians, as well as to discuss the possible policy implications. METHODS: Based on data from the 2004 and 2006 National Survey of Physicians, Dentists and Pharmacists, we estimated the future supply of physicians by specialty, using multistate life tables. Based on possible scenarios of the future increase in female physicians, we also estimated the supply of physicians by specialty. RESULTS: Even if Japan's current medical school enrollment capacity is maintained in subsequent years, the number of physicians per 1000 population is expected to increase from 2.2 in 2006 to 3.2 in 2036, which is a 46% increase from the current level. The numbers of obstetrician/gynecologists (OB/GYNs) and surgeons are expected to temporarily decline from their current level, whereas the number of OB/GYNs per 1000 births will still increase because of the declining number of births. The number of surgeons per 1000 population, even with the decreasing population, will decline temporarily over the next few years. If the percentage of female physicians continues to increase, the overall number of physicians will not be significantly affected, but in specialties with current very low female physician participation rates, such as surgery, the total number of physicians is expected to decline significantly. CONCLUSION: At the current medical school enrollment capacity, the number of physicians per population is expected to continue to increase because of the skewed age distribution of physicians and the declining population in Japan. However, with changes in young physicians' choices of medical specialties and as the percentage of female physicians increases, patterns of physician supply will vary between specialties. Specialties less often chosen by young physicians and where males have dominated will face a decline in physician supply. These results highlight the necessity for developing a work environment that attracts female physicians to these types of specialties. This will also lead to improved gender equality in the workforce and more effective use of human resources. PMID- 19811626 TI - A vision of the future for BMC Medicine: serving science, medicine and authors. AB - In June 2009, BMC Medicine received its first official impact factor of 3.28 from Thomson Reuters. In recognition of this landmark event, the BMC Medicine editorial team present and discuss the vision and aims of the journal. PMID- 19811627 TI - Retroperitoneal abscess complicated with necrotizing fasciitis of the thigh in a patient with sigmoid colon cancer. AB - BACKGROUND: Necrotizing fasciitis of the thigh due to the colon cancer, especially during chemotherepy, has not been previously reported. CASE PRESENTATION: A 67-year-old man admitted to the hospital was diagnosed with sigmoid colon cancer that had spread to the left psoas muscle. Multiple hepatic metastases were also found, and combination chemotherapy with irinotecan and S-1 was administered. Four months after the initiation of chemotherapy, the patient developed gait disturbance and high fever and was therefore admitted to the emergency department of our hospital. Blood examination revealed generalized inflammation with a high C-reactive protein level. Computed tomography of the abdomen and pelvis showed gas and fluid collection in the retroperitoneum adjacent to the sigmoid colon cancer. The abscess was locally drained under computed tomographic guidance; however, the infection continued to spread and necrotizing fasciitis developed. Consequently, emergent debridement was performed. The patient recovered well, and the primary tumor was resected after remission of the local inflammation. CONCLUSION: Necrotizing fasciitis of the thigh due to the spread of sigmoid colon cancer is unusual, but this fatal complication should be considered during chemotherapy for patients with unresectable colorectal cancer. PMID- 19811628 TI - Spatial and temporal investigations of reported movements, births and deaths of cattle and pigs in Sweden. AB - BACKGROUND: Livestock movements can affect the spread and control of contagious diseases and new data recording systems enable analysis of these movements. The results can be used for contingency planning, modelling of disease spread and design of disease control programs. METHODS: Data on the Swedish cattle and pig populations during the period July 2005 until June 2006 were obtained from databases held by the Swedish Board of Agriculture. Movements of cattle and pigs were investigated from geographical and temporal perspectives, births and deaths of cattle were investigated from a temporal perspective and the geographical distribution of holdings was also investigated. RESULTS: Most movements of cattle and pigs were to holdings within 100 km, but movements up to 1200 km occurred. Consequently, the majority of movements occurred within the same county or to adjacent counties. Approximately 54% of the cattle holdings and 45% of the pig holdings did not purchase any live animals. Seasonal variations in births and deaths of cattle were identified, with peaks in spring. Cattle movements peaked in spring and autumn. The maximum number of holdings within a 3 km radius of one holding was 45 for cattle and 23 for pigs, with large variations among counties. Missing data and reporting bias (digit preference) were detected in the data. CONCLUSION: The databases are valuable tools in contact tracing. However since movements can be reported up to a week after the event and some data are missing they cannot replace other methods in the acute phase of an outbreak. We identified long distance transports of cattle and pigs, and these findings support an implementation of a total standstill in the country in the case of an outbreak of foot-and-mouth disease. The databases contain valuable information and improvements in data quality would make them even more useful. PMID- 19811629 TI - Reduction of liver macrophage transduction by pseudotyping lentiviral vectors with a fusion envelope from Autographa californica GP64 and Sendai virus F2 domain. AB - BACKGROUND: Lentiviral vectors are well suited for gene therapy because they can mediate long-term expression in both dividing and nondividing cells. However, lentiviral vectors seem less suitable for liver gene therapy because systemically administered lentiviral vectors are preferentially sequestered by liver macrophages. This results in a reduction of available virus and might also increase the immune response to the vector and vector products.Reduction of macrophage sequestration is therefore essential for efficient lentiviral liver gene therapy. RESULTS: Fusions were made of Autographa californica GP64 and the hepatocyte specific Sendai Virus envelope proteins. Lentiviral vectors were produced with either wild type GP64, Sendai-GP64, or both wild type GP64 and Sendai-GP64 and tested in vitro and in vivo for hepatocyte and macrophage gene transfer.Sendai-GP64 pseudotyped vectors showed specific gene transfer to HepG2 hepatoma cells, with no detectable transduction of HeLa cervical carcinoma cells, and a decreased affinity for RAW mouse macrophages. Co-expression of wild type GP64 and Sendai-GP64 resulted in improved viral titers while retaining increased affinity for HepG2 cells.In vivo, the Sendai-GP64 vectors also showed decreased transduction of murine liver macrophages. CONCLUSION: We demonstrate reduced macrophage transduction in vitro and in vivo with GP64/Sendai chimeric envelope proteins. PMID- 19811630 TI - Patients' experiences of living with and receiving treatment for fibromyalgia syndrome: a qualitative study. AB - BACKGROUND: Fibromyalgia syndrome (FMS) presents a challenge for patients and health care staff across many medical specialities. The aetiology is multi dimensional, involving somatic, psychological and social factors. Patients' views were obtained to understand their experience of living with this long-term condition, using qualitative interviews. METHODS: 12 patients were recruited and stratified by age, gender and ethnicity from one rheumatology outpatient clinic, and a departmental held database of patients diagnosed with FMS. RESULTS: Patients' accounts of their experience of FMS resonated well with two central concepts: social identity and illness intrusiveness. These suggested three themes for the analytical framework: life before and after diagnosis (e.g. lack of information about FMS, invisibility of FMS); change in health identity (e.g. mental distress, impact on social life) and perceived quality of care (e.g. lack of contact with nurses, attitudes of specialists). The information provided from one male participant did not differ from the female patients, but black and ethnic community patients expressed a degree of suspicion towards the medication prescribed, and the attitudes displayed by some doctors, a finding that has not been previously reported amongst this patient group. Patients expected more consultation time and effective treatment than they received. Subjective experiences and objective physical and emotional changes were non-overlapping. Patients' accounts revealed that their physical, mental and social health was compromised, at times overwhelming and affected their identity. CONCLUSION: FMS is a condition that intrudes upon many aspects of patients' lives and is little understood. At the same time, it is a syndrome that evokes uneasiness in health care staff (as current diagnostic criteria are not well supported by objective markers of physiological or biochemical nature, and indeed because of doubt about the existence of the condition) and places great demands on resources in clinical practice. Greater attention needs to be paid to the links between the explanatory models of patients and staff, and most important, to the interrelationship between the complex physical, psychological and social needs of patients with FMS. Taking a less medical but more holistic approach when drawing up new diagnostic criteria for FMS might match better individuals' somatic and psycho social symptom profile and may result in more effective treatment. PMID- 19811631 TI - Occurrence of Mycobacterium avium subspecies paratuberculosis across host species and European countries with evidence for transmission between wildlife and domestic ruminants. AB - BACKGROUND: Mycobacterium avium subspecies paratuberculosis (Map) causes an infectious chronic enteritis (paratuberculosis or Johne's disease) principally of ruminants. The epidemiology of Map is poorly understood, particularly with respect to the role of wildlife reservoirs and the controversial issue of zoonotic potential (Crohn's disease). Genotypic discrimination of Map isolates is pivotal to descriptive epidemiology and resolving these issues. This study was undertaken to determine the genetic diversity of Map, enhance our understanding of the host range and distribution and assess the potential for interspecies transmission. RESULTS: 164 Map isolates from seven European countries representing 19 different host species were genotyped by standardized IS900- restriction fragment length polymorphism (IS900-RFLP), pulsed-field gel electrophoresis (PFGE), amplified fragment length polymorphisms (AFLP) and mycobacterial interspersed repeat unit-variable number tandem repeat (MIRU-VNTR) analyses. Six PstI and 17 BstEII IS900-RFLP, 31 multiplex [SnaBI-SpeI] PFGE profiles and 23 MIRU-VNTR profiles were detected. AFLP gave insufficient discrimination of isolates for meaningful genetic analysis. Point estimates for Simpson's index of diversity calculated for the individual typing techniques were in the range of 0.636 to 0.664 but a combination of all three methods increased the discriminating power to 0.879, sufficient for investigating transmission dynamics. Two predominant strain types were detected across Europe with all three typing techniques. Evidence for interspecies transmission between wildlife and domestic ruminants on the same property was demonstrated in four cases, between wildlife species on the same property in two cases and between different species of domestic livestock on one property. CONCLUSION: The results of this study showed that it is necessary to use multiple genotyping techniques targeting different sources of genetic variation to obtain the level of discrimination necessary to investigate transmission dynamics and trace the source of Map infections. Furthermore, the combination of genotyping techniques may depend on the geographical location of the population to be tested. Identical genotypes were obtained from Map isolated from different host species co-habiting on the same property strongly suggesting that interspecies transmission occurs. Interspecies transmission of Map between wildlife species and domestic livestock on the same property provides further evidence to support a role for wildlife reservoirs of infection. PMID- 19811632 TI - Generation of Gene Ontology benchmark datasets with various types of positive signal. AB - BACKGROUND: The analysis of over-represented functional classes in a list of genes is one of the most essential bioinformatics research topics. Typical examples of such lists are the differentially expressed genes from transcriptional analysis which need to be linked to functional information represented in the Gene Ontology (GO). Despite the importance of this procedure, there is a little work on consistent evaluation of various GO analysis methods. Especially, there is no literature on creating benchmark datasets for GO analysis tools. RESULTS: We propose a methodology for the evaluation of GO analysis tools, which consists of creating gene lists with a selected signal level and a selected number of independent over-represented classes. The methodology starts with a real life GO data matrix, and therefore the generated datasets have similar features to real positive datasets. The user can select the signal level for over representation, the number of independent positive classes in the dataset, and the size of the final gene list. We present the use of the effective number and various normalizations while embedding the signal to a selected class or classes and the use of binary correlation to ensure that the selected signal classes are independent with each other. The usefulness of generated datasets is demonstrated by comparing different GO class ranking and GO clustering methods. CONCLUSION: The presented methods aid the development and evaluation of GO analysis methods as they enable thorough testing with different signal types and different signal levels. As an example, our comparisons reveal clear differences between compared GO clustering and GO de-correlation methods. The implementation is coded in Matlab and is freely available at the dedicated website http://ekhidna.biocenter.helsinki.fi/users/petri/public/POSGODA/POSGODA.html. PMID- 19811633 TI - Expressions of ECE-CYC2 clade genes relating to abortion of both dorsal and ventral stamens in Opithandra (Gesneriaceae). AB - BACKGROUND: ECE-CYC2 clade genes known in patterning floral dorsoventral asymmetry (zygomorphy) in Antirrhinum majus are conserved in the dorsal identity function including arresting the dorsal stamen. However, it remains uncertain whether the same mechanism underlies abortion of the ventral stamens, an important morphological trait related to evolution and diversification of zygomorphy in Lamiales sensu lato, a major clade of predominantly zygomorphically flowered angiosperms. Opithandra (Gesneriaceae) is of particular interests in addressing this question as it is in the base of Lamiales s.l., an early representative of this type zygomorphy. RESULTS: We investigated the expression patterns of four ECE-CYC2 clade genes and two putative target cyclinD3 genes in Opithandra using RNA in situ hybridization and RT-PCR. OpdCYC gene expressions were correlated with abortion of both dorsal and ventral stamens in Opithandra, strengthened by the negatively correlated expression of their putative target OpdcyclinD3 genes. The complement of OpdcyclinD3 to OpdCYC expressions further indicated that OpdCYC expressions were related to the dorsal and ventral stamen abortion through negative effects on OpdcyclinD3 genes. CONCLUSION: These results suggest that ECE-CYC2 clade TCP genes are not only functionally conserved in the dorsal stamen repression, but also involved in arresting ventral stamens, a genetic mechanism underlying the establishment of zygomorphy with abortion of both the dorsal and ventral stamens evolved in angiosperms, especially within Lamiales s.l. PMID- 19811634 TI - ST3Gal.I sialyltransferase relevance in bladder cancer tissues and cell lines. AB - BACKGROUND: The T antigen is a tumor-associated structure whose sialylated form (the sialyl-T antigen) involves the altered expression of sialyltransferases and has been related with worse prognosis. Since little or no information is available on this subject, we investigated the regulation of the sialyltransferases, able to sialylate the T antigen, in bladder cancer progression. METHODS: Matched samples of urothelium and tumor tissue, and four bladder cancer cell lines were screened for: ST3Gal.I, ST3Gal.II and ST3Gal.IV mRNA level by real-time PCR. Sialyl-T antigen was detected by dot blot and flow cytometry using peanut lectin. Sialyltransferase activity was measured against the T antigen in the cell lines. RESULTS: In nonmuscle-invasive bladder cancers, ST3Gal.I mRNA levels were significantly higher than corresponding urothelium (p < 0.001) and this increase was twice more pronounced in cancers with tendency for recurrence. In muscle-invasive cancers and matching urothelium, ST3Gal.I mRNA levels were as elevated as nonmuscle-invasive cancers. Both non-malignant bladder tumors and corresponding urothelium showed ST3Gal.I mRNA levels lower than all the other specimen groups. A good correlation was observed in bladder cancer cell lines between the ST3Gal.I mRNA level, the ST activity (r = 0.99; p = 0.001) and sialyl-T antigen expression, demonstrating that sialylation of T antigen is attributable to ST3Gal.I. The expression of sialyl-T antigens was found in patients' bladder tumors and urothelium, although without a marked relationship with mRNA level. The two ST3Gal.I transcript variants were also equally expressed, independently of cell phenotype or malignancy. CONCLUSION: ST3Gal.I plays the major role in the sialylation of the T antigen in bladder cancer. The overexpression of ST3Gal.I seems to be part of the initial oncogenic transformation of bladder and can be considered when predicting cancer progression and recurrence. PMID- 19811635 TI - BIRI: a new approach for automatically discovering and indexing available public bioinformatics resources from the literature. AB - BACKGROUND: The rapid evolution of Internet technologies and the collaborative approaches that dominate the field have stimulated the development of numerous bioinformatics resources. To address this new framework, several initiatives have tried to organize these services and resources. In this paper, we present the BioInformatics Resource Inventory (BIRI), a new approach for automatically discovering and indexing available public bioinformatics resources using information extracted from the scientific literature. The index generated can be automatically updated by adding additional manuscripts describing new resources. We have developed web services and applications to test and validate our approach. It has not been designed to replace current indexes but to extend their capabilities with richer functionalities. RESULTS: We developed a web service to provide a set of high-level query primitives to access the index. The web service can be used by third-party web services or web-based applications. To test the web service, we created a pilot web application to access a preliminary knowledge base of resources. We tested our tool using an initial set of 400 abstracts. Almost 90% of the resources described in the abstracts were correctly classified. More than 500 descriptions of functionalities were extracted. CONCLUSION: These experiments suggest the feasibility of our approach for automatically discovering and indexing current and future bioinformatics resources. Given the domain independent characteristics of this tool, it is currently being applied by the authors in other areas, such as medical nanoinformatics. BIRI is available at http://edelman.dia.fi.upm.es/biri/. PMID- 19811636 TI - Health-care associated infections rates, length of stay, and bacterial resistance in an intensive care unit of Morocco: findings of the International Nosocomial Infection Control Consortium (INICC). AB - BACKGROUND: Most studies related to healthcare-associated infection (HAI) were conducted in the developed countries. We sought to determine healthcare associated infection rates, microbiological profile, bacterial resistance, length of stay (LOS), and extra mortality in one ICU of a hospital member of the International Infection Control Consortium (INICC) in Morocco. METHODS: We conducted prospective surveillance from 11/2004 to 4/2008 of HAI and determined monthly rates of central vascular catheter-associated bloodstream infection (CVC BSI), catheter-associated urinary tract infection (CAUTI) and ventilator associated pneumonia (VAP). CDC-NNIS definitions were applied. device-utilization rates were calculated by dividing the total number of device-days by the total number of patient-days. Rates of VAP, CVC-BSI, and CAUTI per 1000 Device-days were calculated by dividing the total number of HAI by the total number of specific Device-days and multiplying the result by 1000. RESULTS: 1,731 patients hospitalized for 11,297 days acquired 251 HAIs, an overall rate of 14.5%, and 22.22 HAIs per 1,000 ICU-days. The central venous catheter-related bloodstream infections (CVC-BSI) rate found was 15.7 per 1000 catheter-days; the ventilator associated pneumonia (VAP) rate found was 43.2 per 1,000 ventilator-days; and the catheter-associated urinary tract infections (CAUTI) rate found was 11.7 per 1,000 catheter-days.Overall 25.5% of all Staphylococcus aureus HAIs were caused by methicillin-resistant strains, 78.3% of Coagulase-negative-staphylococci were methicillin resistant as well. 75.0% of Klebsiella were resistant to ceftriaxone and 69.5% to ceftazidime. 31.9% of E. Coli were resistant to ceftriaxone and 21.7% to ceftazidime. 68.4% of Enterobacter sp were resistant to ceftriaxone, 55.6% to ceftazidime, and 10% to imipenem; 35.6% of Pseudomonas sp were resistant to ceftazidime and 13.5% to imipenem.LOS of patients was 5.1 days for those without HAI, 9.0 days for those with CVC-BSI, 10.6 days for those with VAP, and 13.7 days for those with CAUTI.Extra mortality was 56.7% (RR, 3.28; P =< 0.001) for VAP, 75.1% (RR, 4.02; P = 0.0027) for CVC-BSI, and 18.7% (RR, 1.75; P = 0.0218) for CAUTI. CONCLUSION: HAI rates, LOS, mortality, and bacterial resistance were high. Even if data may not reflect accurately the clinical setting of the country, programs including surveillance, infection control, and antibiotic policy are a priority in Morocco. PMID- 19811637 TI - Phase I dose escalation pharmacokinetic assessment of intravenous humanized anti MUC1 antibody AS1402 in patients with advanced breast cancer. AB - INTRODUCTION: MUC1 is a cell-surface glycoprotein that establishes a molecular barrier at the epithelial surface and engages in morphogenetic signal transduction. Alterations in MUC1 glycosylation accompany the development of cancer and influence cellular growth, differentiation, transformation, adhesion, invasion, and immune surveillance. A 20-amino-acid tandem repeat that forms the core protein of MUC1 is overexpressed and aberrantly glycosylated in the majority of epithelial tumors. AS1402 (formerly R1550) is a humanized IgG1k monoclonal antibody that binds to PDTR sequences within this tandem repeat that are not exposed in normal cells. AS1402 is a potent inducer of antibody-dependent cellular cytotoxicity (ADCC), specifically against MUC1-expressing tumor cells. The objective of this study was to determine the safety, tolerability, and pharmacokinetic (PK) characteristics of AS1402 monotherapy in patients with locally advanced or metastatic MUC1-positive breast cancer that had progressed after anthracyclines- and taxane-based therapy. METHODS: Patients received AS1402 over a 1- to 3-hour intravenous (i.v.) infusion at doses between 1 and 16 mg/kg, with repeated dosing every 1 to 3 weeks (based on patient-individualized PK assessment) until disease progression. Serum AS1402 levels were measured at multiple times after i.v. administration. Human anti-human antibody (HAHA) responses were measured to determine the immunogenicity of AS1402. Noncompartmental pharmacokinetic parameters were determined and were used to assess dose dependency across the dose range studied. RESULTS: Twenty-six patients were treated. AS1402 was generally well tolerated. Two grade 3/4 drug related adverse events were reported, both at the 3-mg/kg dose. Neither was observed in expanded or subsequent dosing cohorts. No anti-human antibodies were detected. Plasma concentrations of AS1402 appeared to be proportional to dose within the 1- to 16-mg/kg dose range assessed, with a mean terminal half-life of 115.4 +/- 37.1 hours. CONCLUSIONS: Repeated iv administration of AS1402 was well tolerated, with a maximum tolerated dose (MTD) exceeding 16 mg/kg, the highest dose administered in this study. The half-life and exposure of AS1402 were such that weekly dosing could achieve plasma concentrations corresponding to the maximal ADCC activity observed in vitro. A phase II study is ongoing to evaluate the clinical activity of AS1402 in patients with advanced breast cancer. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT00096057. PMID- 19811638 TI - Future oriented group training for suicidal patients: a randomized clinical trial. AB - BACKGROUND: In routine psychiatric treatment most clinicians inquire about indicators of suicide risk, but once the risk is assessed not many clinicians systematically focus on suicidal thoughts. This may reflect a commonly held opinion that once the depressive or anxious symptoms are effectively treated the suicidal symptoms will wane. Consequently, many clients with suicidal thoughts do not receive systematic treatment of their suicidal thinking. There are many indications that specific attention to suicidal thinking is necessary to effectively decrease the intensity and recurrence of suicidal thinking. We therefore developed a group training for patients with suicidal thoughts that is easy to apply in clinical settings as an addition to regular treatment and that explicitly focuses on suicidal thinking. We hypothesize that such an additional training will decrease the frequency and intensity of suicidal thinking.We based the training on cognitive behavioural approaches of hopelessness, worrying, and future perspectives, given the theories of Beck, McLeod and others, concerning the lack of positive expectations characteristic for many suicidal patients. In collaboration with each participant in the training individual positive future possibilities and goals were challenged. METHODS/DESIGN: We evaluate the effects of our program on suicide ideation (primary outcome measure). The study is conducted in a regular treatment setting with regular inpatients and outpatients representative for Dutch psychiatric treatment settings. The design is a RCT with two arms: TAU (Treatment as Usual) versus TAU plus the training. Follow up measurements are taken 12 months after the first assessment. DISCUSSION: There is a need for research on the effectiveness of interventions in suicidology, especially RCT's. In our treatment program we combine aspects and interventions that have been proven to be useful in the treatment of suicidal thinking and behavior. TRIAL REGISTRATION: ISRCTN56421759. PMID- 19811639 TI - Aquaporin-6 is expressed along the rat gastrointestinal tract and upregulated by feeding in the small intestine. AB - BACKGROUND: Several aquaporins (a family of integral membrane proteins) have been recently identified in the mammalian gastrointestinal tract, and their involvement in the movement of fluid and small solutes has been suggested. In this direction we investigated, in some regions of the rat gastrointestinal tract, the presence and localization of aquaporin-6, given its peculiar function as an ion selective channel. RESULTS: RT-PCR and immunoblotting experiments showed that aquaporin-6 was expressed in all the investigated portions of the rat gastrointestinal tract. The RT-PCR experiments showed that aquaporin-6 transcript was highly expressed in small intestine and rectum, and less in stomach, caecum and colon. In addition, jejunal mRNA expression was specifically stimulated by feeding. Immunoblotting analysis showed a major band with a molecular weight of about 55 kDa corresponding to the aquaporin-6 protein dimer; this band was stronger in the stomach and large intestine than in the small intestine. Immunoblotting analysis of brush border membrane vesicle preparations showed an intense signal for aquaporin-6 protein. The results of in situ hybridization experiments demonstrate that aquaporin-6 transcript is present in the isthmus, neck and basal regions of the stomach lining, and throughout the crypt-villus axis in both small and large intestine. In the latter regions, immunohistochemistry revealed strong aquaporin-6 labelling in the apical membrane of the surface epithelial cells, while weak or no labelling was observed in the crypt cells. In the stomach, an intense staining was observed in mucous neck cells and lower signal in principal cells and some parietal cells. CONCLUSION: The results indicate that aquaporin-6 is distributed throughout the gastrointestinal tract. Aquaporin-6 localization at the apical pole of the superficial epithelial cells and its upregulation by feeding suggest that it may be involved in movements of water and anions through the epithelium of the villi. PMID- 19811640 TI - Extraction of average neck flexion angle during swallowing in neutral and chin tuck positions. AB - BACKGROUND: A common but debated technique in the management of swallowing difficulties is the chin tuck swallow, where the neck is flexed forward prior to swallowing. Natural variations in chin tuck angles across individuals may contribute to the differential effectiveness of the technique. METHODOLOGY: To facilitate the study of chin tuck angle variations, we present a template tracking algorithm that automatically extracts neck angles from sagittal videos of individuals performing chin tuck swallows. Three yellow markers geometrically arranged on a pair of dark visors were used as tracking cues. RESULTS: The algorithm was applied to data collected from 178 healthy participants during neutral and chin tuck position swallows. Our analyses revealed no major influences of body mass index and age on neck flexion angles during swallowing, while gender influenced the average neck angle only during wet swallows in the neutral position. Chin tuck angles seem to be independent of anthropometry and gender in healthy adults, but deserve further study in pathological populations. CONCLUSION: The proposed neck flexion angle extraction algorithm may be useful in future studies where strict participant compliance to swallowing task protocol can be assured. PMID- 19811641 TI - Discovering multiple realistic TFBS motifs based on a generalized model. AB - BACKGROUND: Identification of transcription factor binding sites (TFBSs) is a central problem in Bioinformatics on gene regulation. de novo motif discovery serves as a promising way to predict and better understand TFBSs for biological verifications. Real TFBSs of a motif may vary in their widths and their conservation degrees within a certain range. Deciding a single motif width by existing models may be biased and misleading. Additionally, multiple, possibly overlapping, candidate motifs are desired and necessary for biological verification in practice. However, current techniques either prohibit overlapping TFBSs or lack explicit control of different motifs. RESULTS: We propose a new generalized model to tackle the motif widths by considering and evaluating a width range of interest simultaneously, which should better address the width uncertainty. Moreover, a meta-convergence framework for genetic algorithms (GAs), is proposed to provide multiple overlapping optimal motifs simultaneously in an effective and flexible way. Users can easily specify the difference amongst expected motif kinds via similarity test. Incorporating Genetic Algorithm with Local Filtering (GALF) for searching, the new GALF-G (G for generalized) algorithm is proposed based on the generalized model and meta-convergence framework. CONCLUSION: GALF-G was tested extensively on over 970 synthetic, real and benchmark datasets, and is usually better than the state-of-the-art methods. The range model shows an increase in sensitivity compared with the single-width ones, while providing competitive precisions on the E. coli benchmark. Effectiveness can be maintained even using a very small population, exhibiting very competitive efficiency. In discovering multiple overlapping motifs in a real liver-specific dataset, GALF-G outperforms MEME by up to 73% in overall F-scores. GALF-G also helps to discover an additional motif which has probably not been annotated in the dataset. http://www.cse.cuhk.edu.hk/%7Etmchan/GALFG/ PMID- 19811642 TI - Medicine and psychiatry in Western culture: Ancient Greek myths and modern prejudices. AB - The origins of Western culture extensively relate to Ancient Greek culture. While many ancient cultures have contributed to our current knowledge about medicine and the origins of psychiatry, the Ancient Greeks were among the best observers of feelings and moods patients expressed towards medicine and toward what today is referred to as 'psychopathology'. Myths and religious references were used to explain what was otherwise impossible to understand or be easily communicated. Most ancient myths focus on ambiguous feelings patients may have had towards drugs, especially psychotropic ones. Interestingly, such prejudices are common even today. Recalling ancient findings and descriptions made using myths could represent a valuable knowledge base for modern physicians, especially for psychiatrists and their patients, with the aim of better understanding each other and therefore achieving a better clinical outcome. This paper explores many human aspects and feelings towards doctors and their cures, referring to ancient myths and focusing on the perception of mental illness. PMID- 19811643 TI - Ability of Group IVB metallocene polyethers containing dienestrol to arrest the growth of selected cancer cell lines. AB - BACKGROUND: Monomeric Group IVB (Ti, Zr and Hf) metallocenes represent a new class of antitumor compounds. There is literature on the general biological activities of some organotin compounds. Unfortunately, there is little information with respect to the molecular level activity of these organotin compounds. We recently started focusing on the anti-cancer activity of organotin polymers that we had made for other purposes and as part of our platinum anti cancer effort. METHODS: For this study, we synthesized a new series of metallocene-containing compounds coupling the metallocene unit with dienestrol, a synthetic, nonsteroidal estrogen. This is part of our effort to couple known moieties that offer antitumor activity with biologically active units hoping to increase the biological activity of the combination. The materials were confirmed to be polymeric using light scattering photometry and the structural repeat unit was verified employing matrix assisted laser desorption ionization mass spectrometry and infrared spectroscopy results. RESULTS: The polymers demonstrated the ability to suppress the growth of a series of tumor cell lines originating from breast, colon, prostrate, and lung cancers at concentrations generally lower than those required for inhibition of cell growth by the commonly used antitumor drug cisplatin. CONCLUSION: These drugs show great promise in vitro against a number of cancer cell lines and due to their polymeric nature will most likely be less toxic than currently used metal-containing drugs such as cisplatin. These drugs also offer several addition positive aspects. First, the reactants are commercially available so that additional synthetic steps are not needed. Second, synthesis of the polymer is rapid, occurring within about 15 seconds. Third, the interfacial synthetic system is already industrially employed in the synthesis of aromatic nylons and polycarbonates. Thus, the ability to synthesize large amounts of the drugs is straight forward. PMID- 19811644 TI - Management of a massive thoracoabdominal impalement: a case report. AB - A 26 year old male was impaled through his chest and upper abdomen with an iron angle, one and half meter long and five centimeters thick. The iron angle entered the chest, through the epigastrium and exited posteriorly just inferior to the angle of left scapula. The patient was transported to hospital with the iron angle in situ. Positioning the patient for intubation proved a major challenge. An unconventional position for intubation allowed a successful airway management. Paucity of time prevented us from gauging the nature and extent of injury. The challenges posed by massive impalement could be successfully managed due to rapid pre-hospital transfer and co-ordinated team effort. PMID- 19811645 TI - Hedgehog pathway responsiveness correlates with the presence of primary cilia on prostate stromal cells. AB - BACKGROUND: Hedgehog (Hh) signaling from the urogenital sinus (UGS) epithelium to the surrounding mesenchyme plays a critical role in regulating ductal formation and growth during prostate development. The primary cilium, a feature of most interphase vertebrate cell types, serves as a required localization domain for Hh signaling transducing proteins. RESULTS: Immunostaining revealed the presence of primary cilia in mesenchymal cells of the developing prostate. Cell-based assays of a urongenital sinus mesenchymal cell line (UGSM-2) revealed that proliferation limiting (serum starvation and/or confluence) growth conditions promoted cilia formation and correlated with pathway activation associated with accumulation of Smoothened in primary cilia. The prostate cancer cell lines PC-3, LNCaP, and 22RV1, previously shown to lack demonstrable autocrine Hh signaling capacity, did not exhibit primary cilia even under proliferation-limiting growth conditions. CONCLUSION: We conclude that paracrine Hedgehog signaling activity in the prostate is associated with the presence of primary cilia on stromal cells but that a role in autocrine Hh signaling remains speculative. PMID- 19811646 TI - An occasional diagnosis of myasthenia gravis--a focus on thymus during cardiac surgery: a case report. AB - BACKGROUND: Myasthenia gravis, an uncommon autoimmune syndrome, is commonly associated with thymus abnormalities. Thymomatous myasthenia gravis is considered to have worst prognosis and thymectomy can reverse symptoms if precociously performed. CASE REPORT: We describe a case of a patient who underwent mitral valve repair and was found to have an occasional thymomatous mass during the surgery. A total thymectomy was performed concomitantly to the mitral valve repair. CONCLUSION: The diagnosis of thymomatous myasthenia gravis was confirmed postoperatively. Following the surgery this patient was strictly monitored and at 1-year follow-up a complete stable remission had been successfully achieved. PMID- 19811647 TI - Whole genome single nucleotide polymorphism based phylogeny of Francisella tularensis and its application to the development of a strain typing assay. AB - BACKGROUND: A low genetic diversity in Francisella tularensis has been documented. Current DNA based genotyping methods for typing F. tularensis offer a limited and varying degree of subspecies, clade and strain level discrimination power. Whole genome sequencing is the most accurate and reliable method to identify, type and determine phylogenetic relationships among strains of a species. However, lower cost typing schemes are necessary in order to enable typing of hundreds or even thousands of isolates. RESULTS: We have generated a high-resolution phylogenetic tree from 40 Francisella isolates, including 13 F. tularensis subspecies holarctica (type B) strains, 26 F. tularensis subsp. tularensis (type A) strains and a single F. novicida strain. The tree was generated from global multi-strain single nucleotide polymorphism (SNP) data collected using a set of six Affymetrix GeneChip resequencing arrays with the non repetitive portion of LVS (type B) as the reference sequence complemented with unique sequences of SCHU S4 (type A). Global SNP based phylogenetic clustering was able to resolve all non-related strains. The phylogenetic tree was used to guide the selection of informative SNPs specific to major nodes in the tree for development of a genotyping assay for identification of F. tularensis subspecies and clades. We designed and validated an assay that uses these SNPs to accurately genotype 39 additional F. tularensis strains as type A (A1, A2, A1a or A1b) or type B (B1 or B2). CONCLUSION: Whole-genome SNP based clustering was shown to accurately identify SNPs for differentiation of F. tularensis subspecies and clades, emphasizing the potential power and utility of this methodology for selecting SNPs for typing of F. tularensis to the strain level. Additionally, whole genome sequence based SNP information gained from a representative population of strains may be used to perform evolutionary or phylogenetic comparisons of strains, or selection of unique strains for whole-genome sequencing projects. PMID- 19811648 TI - Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations. AB - BACKGROUND: The establishment of mutant populations together with the strategies for targeted mutation detection has been applied successfully to a large number of organisms including many species in the plant kingdom. Considerable efforts have been invested into research on tomato as a model for berry-fruit plants. With the progress of the tomato sequencing project, reverse genetics becomes an obvious and achievable goal. RESULTS: Here we describe the treatment of Solanum lycopersicum seeds with 1% EMS and the development of a new mutated tomato population. To increase targeted mutant detection throughput an automated seed DNA extraction has been combined with novel mutation detection platforms for TILLING in plants. We have adapted two techniques used in human genetic diagnostics: Conformation Sensitive Capillary Electrophoresis (CSCE) and High Resolution DNA Melting Analysis (HRM) to mutation screening in DNA pools. Classical TILLING involves critical and time consuming steps such as endonuclease digestion reactions and gel electrophoresis runs. Using CSCE or HRM, the only step required is a simple PCR before either capillary electrophoresis or DNA melting curve analysis. Here we describe the development of a mutant tomato population, the setting up of two polymorphism detection platforms for plants and the results of the first screens as mutation density in the populations and estimation of the false-positives rate when using HRM to screen DNA pools. CONCLUSION: These results demonstrate that CSCE and HRM are fast, affordable and sensitive techniques for mutation detection in DNA pools and therefore allow the rapid identification of new allelic variants in a mutant population. Results from the first screens indicate that the mutagen treatment has been effective with an average mutation detection rate per diploid genome of 1.36 mutation/kb/1000 lines. PMID- 19811649 TI - Placental malaria and low birth weight in pregnant women living in a rural area of Burkina Faso following the use of three preventive treatment regimens. AB - BACKGROUND: The weekly chemoprophylaxis of malaria during pregnancy with chloroquine (CQ) has become problematic with the increasing resistance of Plasmodium falciparum to this drug. There was a need to test the benefits of new strategies over the classical chemoprophylaxis. This study was conducted to provide data to the National Malarial Control Programme for an evidence-based policy change decision making process. It compares the efficacy of two IPT regimens, using chloroquine (CQ) or sulphadoxine/pyrimethamine (SP), with the classical chemoprophylaxis regimen using CQ in reducing the adverse outcomes of malaria infection, for the mother and the foetus. METHODS: Pregnant women attending the first antenatal care visit were randomly assigned to one of the three treatment regimens. They were subsequently followed up till delivery. Maternal, placental and cord blood samples were obtained upon delivery to check for P. falciparum infection. RESULTS: A total of 648 pregnant women were enrolled in the study. Delivery outcome were available for 423 of them. Peripheral maternal P. falciparum infection at delivery was found in 25.8% of the women. The proportion of women with maternal infection was significantly lower in the IPTp/SP group than in the CQ group (P << 0.000). The prevalence of placental malaria was 18.8% in the CWC/CQ group; 15.9% in the IPTp/CQ group and 10.6% in the IPTp/SP group. The incidence of LBW (weight < 2,500 g) was significantly higher among infants of mothers in the CWC/CQ group (23.9%) as compared with those of mothers in the IPTp/CQ (15.6%) and IPTp/SP (11.6%) groups (p = 0.02) CONCLUSION: Intermittent preventive treatment with SP has shown clear superiority in reducing adverse outcomes at delivery, as compared with intermittent preventive treatment with CQ and classical chemoprophylaxis with CQ. PMID- 19811650 TI - Miscarriage rates after dehydroepiandrosterone (DHEA) supplementation in women with diminished ovarian reserve: a case control study. AB - BACKGROUND: Dehydroepinadrosterone (DHEA) supplementation improves pregnancy chances in women with diminished ovarian reserve (DOR), by possibly reducing aneuploidy. Since a large majority of spontaneous miscarriages are associated with aneuploidy, one can speculate that DHEA supplementation may also reduce miscarriage rates. METHODS: We retroactively compared, utilizing two independent statistical models, miscarriage rates in 73 DHEA supplemented pregnancies at two independent North American infertility centers, age-stratified, to miscarriages reported in a national U.S. in vitro fertilization (IVF) data base. RESULTS: After DHEA supplementation the miscarriage rate at both centers was 15.1% (15.0% and 15.2%, respectively). For DHEA supplementation Mantel-Hanszel common odds ratio (and 95% confidence interval), stratified by age, was significantly lower, relative to odds of miscarriage in the general IVF control population [0.49 (0.25 0.94; p = 0.04)]. Miscarriage rates after DHEA were significantly lower at all ages but most pronounced above age 35 years. DISCUSSION: Since DOR patients in the literature are reported to experience significantly higher miscarriage rates than average IVF patients, the here observed reduction in miscarriages after DHEA supplementation exceeds, however, all expectations. Miscarriage rates after DHEA not only were lower than in an average national IVF population but were comparable to rates reported in normally fertile populations. Low miscarriage rates, comparable to those of normal fertile women, are statistically impossible to achieve in DOR patients without assumption of a DHEA effect on embryo ploidy. Beyond further investigations in infertile populations, these data, therefore, also suggest the investigations of pre-conception DHEA supplementation in normal fertile populations above age 35 years. PMID- 19811651 TI - Testing mitochondrial sequences and anonymous nuclear markers for phylogeny reconstruction in a rapidly radiating group: molecular systematics of the Delphininae (Cetacea: Odontoceti: Delphinidae). AB - BACKGROUND: Many molecular phylogenetic analyses rely on DNA sequence data obtained from single or multiple loci, particularly mitochondrial DNA loci. However, phylogenies for taxa that have undergone recent, rapid radiation events often remain unresolved. Alternative methodologies for discerning evolutionary relationships under these conditions are desirable. The dolphin subfamily Delphininae is a group that has likely resulted from a recent and rapid radiation. Despite several efforts, the evolutionary relationships among the species in the subfamily remain unclear. RESULTS: Here, we compare a phylogeny estimated using mitochondrial DNA (mtDNA) control region sequences to a multi locus phylogeny inferred from 418 polymorphic genomic markers obtained from amplified fragment length polymorphism (AFLP) analysis. The two sets of phylogenies are largely incongruent, primarily because the mtDNA tree provides very poor resolving power; very few species' nodes in the tree are supported by bootstrap resampling. The AFLP phylogeny is considerably better resolved and more congruent with relationships inferred from morphological data. Both phylogenies support paraphyly for the genera Stenella and Tursiops. The AFLP data indicate a close relationship between the two spotted dolphin species and recent ancestry between Stenella clymene and S. longirostris. The placement of the Lagenodelphis hosei lineage is ambiguous: phenetic analysis of the AFLP data is consistent with morphological expectations but the phylogenetic analysis is not. CONCLUSION: For closely related, recently diverged taxa, a multi-locus genome-wide survey is likely the most comprehensive approach currently available for phylogenetic inference. PMID- 19811652 TI - ERG-associated protein with SET domain (ESET)-Oct4 interaction regulates pluripotency and represses the trophectoderm lineage. AB - BACKGROUND: Pluripotency, the capacity for indefinite self-renewal and differentiation into diverse cell types is a unique state exhibited by embryonic stem (ES) cells. Transcriptional regulators, such as Oct4, are critical for pluripotency, but the role of epigenetic modifiers remains to be fully elucidated. RESULTS: Here, we show that ERG-associated protein with SET domain (ESET), a histone methyltransferase enzyme, maintains pluripotency through repression of Cdx2, a key trophectoderm determinant, by histone H3 lysine 9 trimethylation (H3K9me3) of the promoter region. Notably, this repression is mediated through the synergistic function of small ubiquitin-related modifier (SUMO)ylated ESET and Oct4. ESET localises to the promyelocytic leukaemia (PML) nuclear bodies and is SUMOylated in ES cells. Interaction of ESET with Oct4 depends on a SUMO-interacting motif (SIM) in Oct4, which is critical for the repression of Cdx2. CONCLUSION: Loss of ESET or Oct4 results in strikingly similar phenotypes both in ES cells with their differentiation into trophectoderm cells, and in early embryos where there is a failure of development of the pluripotent inner cell mass (ICM) of blastocysts. We propose that SUMOylated ESET Oct4 complex is critical for both the initiation and maintenance of pluripotency through repression of differentiation, particularly of the trophectoderm lineage by epigenetic silencing of Cdx2. PMID- 19811653 TI - Reports of evidence planting by police among a community-based sample of injection drug users in Bangkok, Thailand. AB - BACKGROUND: Drug policy in Thailand has relied heavily on law enforcement-based approaches. Qualitative reports indicate that police in Thailand have resorted to planting drugs on suspected drug users to extort money or provide grounds for arrest. The present study sought to describe the prevalence and factors associated with this form of evidence planting by police among injection drug users (IDU) in Bangkok. METHODS: Multivariate logistic regression was used to identify factors associated with evidence planting of drugs by police among a community-based sample of IDU in Bangkok. We also examined the prevalence and average amount of money paid by IDU to police in order to avoid arrest. RESULTS: 252 IDU were recruited between July and August, 2008, among whom 66 (26.2%) were female and the median age was 36.5 years. In total, 122 (48.4%) participants reported having drugs planted on them by police. In multivariate analyses, this form of evidence planting was positively associated with midazolam use (Adjusted Odds Ratio [AOR] = 2.84; 95% Confidence Interval [CI]: 1.58 - 5.11), recent non fatal overdose (AOR = 2.56; 95%CI: 1.40 - 4.66), syringe lending (AOR = 2.08; 95%CI: 1.19 - 3.66), and forced drug treatment (AOR = 1.88; 95%CI: 1.05 - 3.36). Among those who reported having drugs planted on them, 59 (48.3%) paid police a bribe in order to avoid arrest. CONCLUSION: A high proportion of community recruited IDU participating in this study reported having drugs planted on them by police. Drug planting was found to be associated with numerous risk factors including syringe sharing and participation in government-run drug treatment programs. Immediate action should be taken to address this form of abuse of power reportedly used by police. PMID- 19811654 TI - Molecular models for intrastrand DNA G-quadruplexes. AB - BACKGROUND: Independent surveys of human gene promoter regions have demonstrated an overrepresentation of G(3)X(n1)G3X(n2)G(3)X(n3)G(3) motifs which are known to be capable of forming intrastrand quadruple helix structures. In spite of the widely recognized importance of G-quadruplex structures in gene regulation and growing interest around this unusual DNA structure, there are at present only few such structures available in the Nucleic Acid Database. In the present work we generate by molecular modeling feasible G-quadruplex structures which may be useful for interpretation of experimental data. RESULTS: We have used all quadruplex DNA structures deposited in the Nucleic Acid Database in order to select a list of fragments entailing a strand of three adjacent G's paired with another strand of three adjacent G's separated by a loop of one to four residues. These fragments were further clustered and representative fragments were finally selected. Further fragments were generated by assemblying the two strands of each fragment with loops from different fragments whenever the anchor G's were superimposable. The fragments were used to assemble G quadruplex based on a superimposability criterion. CONCLUSION: Molecular models have been generated for a large number of G(3)X(n1)G(3)X(n2)G3X(n3)G(3) sequences. For a given sequence not all topologies are possible with the available repertoire of fragments due to steric hindrance and low superimposability. Since all molecular models are generated by fragments coming from observed quadruplex structures, molecular models are in principle reliable and may be used for interpretation of experimental data. Some examples of applications are given. PMID- 19811655 TI - Detection of NP, N3 and N7 antibodies to avian influenza virus by indirect ELISA using yeast-expressed antigens. AB - BACKGROUND: Avian influenza viruses, belonging to the family Orthomyxoviridae, possess distinct combinations of hemagglutinin (H) and the neuraminidase (N) surface glycoproteins. Typing of both H and N antigens is essential for the epidemiological and surveillance studies. Therefore, it is important to find a rapid, sensitive, and specific method for their assay, and ELISA can be useful for this purpose, by using recombinant proteins. RESULTS: The nucleoprotein (NP) and truncated neuraminidase subtype 3 and 7 of avian influenza virus (AIV) were expressed in Saccharomyces cerevisiae and used to develop an indirect enzyme linked immunosorbent assay for antibody detection. The developed assays were evaluated with a panel of 64 chicken serum samples. The performance of NP-ELISA was compared with the commercially available ProFlok AIV ELISA kit. The results showed comparable agreement and sensitivity between the two tests, indicating that NP-ELISA assay can be used for screening the influenza type A antibody in AIV infected birds. The N3 and N7- ELISAs also reacted specifically to their type specific sera and did not exhibit any cross-reaction with heterologous neuraminidase subtype specific sera. CONCLUSION: The study demonstrates the expression of the NP, N3, and N7 proteins of AIV in yeast (S. cerevisiae) and their application in developing an indirect ELISA for detecting NP, N3 and N7 antibodies from AIV-infected chicken sera. The described indirect ELISAs are rapid, sensitive, specific and can be used as promising tests during serological surveillance. PMID- 19811656 TI - Six host range variants of the xenotropic/polytropic gammaretroviruses define determinants for entry in the XPR1 cell surface receptor. AB - BACKGROUND: The evolutionary interactions between retroviruses and their receptors result in adaptive selection of restriction variants that can allow natural populations to evade retrovirus infection. The mouse xenotropic/polytropic (X/PMV) gammaretroviruses rely on the XPR1 cell surface receptor for entry into host cells, and polymorphic variants of this receptor have been identified in different rodent species. RESULTS: We screened a panel of X/PMVs for infectivity on rodent cells carrying 6 different XPR1 receptor variants. The X/PMVs included 5 well-characterized laboratory and wild mouse virus isolates as well as a novel cytopathic XMV-related virus, termed Cz524, isolated from an Eastern European wild mouse-derived strain, and XMRV, a xenotropic-like virus isolated from human prostate cancer. The 7 viruses define 6 distinct tropisms. Cz524 and another wild mouse isolate, CasE#1, have unique species tropisms. Among the PMVs, one Friend isolate is restricted by rat cells. Among the XMVs, two isolates, XMRV and AKR6, differ from other XMVs in their PMV like restriction in hamster cells. We generated a set of Xpr1 mutants and chimeras, and identified critical amino acids in two extracellular loops (ECLs) that mediate entry of these different viruses, including 3 residues in ECL3 that are involved in PMV entry (E500, T507, and V508) and can also influence infectivity by AKR6 and Cz524. CONCLUSION: We used a set of natural variants and mutants of Xpr1 to define 6 distinct host range variants among naturally occurring X/PMVs (2 XMV variants, 2 PMVs, 2 different wild mouse variants). We identified critical amino acids in XPR1 that mediate entry of these viruses. These gammaretroviruses and their XPR1 receptor are thus highly functionally polymorphic, a consequence of the evolutionary pressures that favor both host resistance and virus escape mutants. This variation accounts for multiple naturally occurring virus resistance phenotypes and perhaps contributes to the widespread distribution of these viruses in rodent and non-rodent species. PMID- 19811657 TI - Detailed analysis of X chromosome inactivation in a 49,XXXXX pentasomy. AB - BACKGROUND: Pentasomy X (49,XXXXX) has been associated with a severe clinical condition, presumably resulting from failure or disruption of X chromosome inactivation. Here we report that some human X chromosomes from a patient with 49,XXXXX pentasomy were functionally active following isolation in inter-specific (human-rodent) cell hybrids. A comparison with cytogenetic and molecular findings provided evidence that more than one active X chromosome was likely to be present in the cells of this patient, accounting for her abnormal phenotype. RESULTS: 5 bromodeoxyuridine (BrdU)-pulsed cultures showed different patterns among late replicating X chromosomes suggesting that their replication was asynchronic and likely to result in irregular inactivation. Genotyping of the proband and her mother identified four maternal and one paternal X chromosomes in the proband. It also identified the paternal X chromosome haplotype (P), indicating that origin of this X pentasomy resulted from two maternal, meiotic non-disjunctions. Analysis of the HUMANDREC region of the androgen receptor (AR) gene in the patient's mother showed a skewed inactivation pattern, while a similar analysis in the proband showed an active paternal X chromosome and preferentially inactivated X chromosomes carrying the 173 AR allele. Analyses of 33 cell hybrid cell lines selected in medium containing hypoxanthine, aminopterin and thymidine (HAT) allowed for the identification of three maternal X haplotypes (M1, M2 and MR) and showed that X chromosomes with the M1, M2 and P haplotypes were functionally active. In 27 cell hybrids in which more than one X haplotype were detected, analysis of X inactivation patterns provided evidence of preferential inactivation. CONCLUSION: Our findings indicated that 12% of X chromosomes with the M1 haplotype, 43.5% of X chromosomes with the M2 haplotype, and 100% of the paternal X chromosome (with the P haplotype) were likely to be functionally active in the proband's cells, a finding indicating that disruption of X inactivation was associated to her severe phenotype. PMID- 19811658 TI - GITR signaling potentiates airway hyperresponsiveness by enhancing Th2 cell activity in a mouse model of asthma. AB - BACKGROUND: Allergic asthma is characterized by airway hyperresponsiveness (AHR) and allergic inflammation of the airways, driven by allergen-specific Th2 cells. The asthma phenotypes and especially AHR are sensitive to the presence and activity of regulatory T (Treg) cells in the lung. Glucocorticoid-induced tumor necrosis factor receptor (GITR) is known to have a co-stimulatory function on effector CD4+ T cells, rendering these cells insensitive to Treg suppression. However, the effects of GITR signaling on polarized Th1 and Th2 cell effector functions are not well-established. We sought to evaluate the effect of GITR signaling on fully differentiated Th1 and Th2 cells and to determine the effects of GITR activation at the time of allergen provocation on AHR and airway inflammation in a Th2-driven mouse model of asthma. METHODS: CD4+CD25- cells were polarized in vitro into Th1 and Th2 effector cells, and re-stimulated in the presence of GITR agonistic antibodies to assess the effect on IFNgamma and IL-4 production. To evaluate the effects of GITR stimulation on AHR and allergic inflammation in a mouse asthma model, BALB/c mice were sensitized to OVA followed by airway challenges in the presence or absence of GITR agonist antibodies. RESULTS: GITR engagement potentiated cytokine release from CD3/CD28-stimulated Th2 but not Th1 cells in vitro. In the mouse asthma model, GITR triggering at the time of challenge induced enhanced airway hyperresponsiveness, serum IgE and ex vivo Th2 cytokine release, but did not increase BAL eosinophilia. CONCLUSION: GITR exerts a differential effect on cytokine release of fully differentiated Th1 and Th2 cells in vitro, potentiating Th2 but not Th1 cytokine production. This effect on Th2 effector functions was also observed in vivo in our mouse model of asthma, resulting in enhanced AHR, serum IgE responses and Th2 cytokine production. This is the first report showing the effects of GITR activation on cytokine production by polarized primary Th1 and Th2 populations and the relevance of this pathway for AHR in mouse models for asthma. Our data provides crucial information on the mode of action of the GITR signaling, a pathway which is currently being considered for therapeutic intervention. PMID- 19811659 TI - Chromophobe renal cell cancer--review of the literature and potential methods of treating metastatic disease. AB - Chromophobe renal cell carcinoma (ChRCC) is a subtype of renal cell carcinoma (RCC). ChRCC is diagnosed mainly in 6th decade of life. An incidence of ChRCC is similar in both men and woman. Eighty six percent of ChRCCs cases are diagnosed in stage 1 or 2. Prognosis of ChRCC is better than in other types of RCC. Five- and 10-year disease free survival (DFS) for ChRCC was 83.9% and 77.9%, respectively. Expression of immunohistological markers: cytokeratins (CK), vimentin, epithelial membrane antigen (EMA), CD10 could be potentially helpful in diagnosis of different subtypes of RCC. From all conventional RCC, CD 117 was detected (overexpression) in membrane of cells ChRCC.Overexpression of CD117 on cellular membranes of ChRCC could be a potential target for kinase inhibitors like: imatinib, dasatinib, nilotinib. The potential targets for other kinase inhibitors (sunitinib and sorafenib) in ChRCC seem to be VEGFR and PDGFR. On the basis for formulating research hypotheses which should be verified by prospective studies. PMID- 19811660 TI - Poly-paraphyly of Hirudinidae: many lineages of medicinal leeches. AB - BACKGROUND: Medicinal leeches became infamous for their utility in bloodletting popularized in the 19th century, and have seen a recent resurgence in post operative treatments for flap and replantation surgeries, and in terms of characterization of salivary anticoagulants. Notorious throughout the world, the quintessential leech family Hirudinidae has been taken for granted to be monophyletic, as has the non-bloodfeeding family Haemopidae. RESULTS: This study is the first to evaluate molecular evidence from hirudinid and haemopid leeches in a manner that encompasses the global scope of their taxonomic distributions. We evaluated the presumed monophyly of the Hirudinidae and assessed previous well accepted classification schemes. The Hirudinidae were found not to be monophyletic, falling instead into two distinct and unrelated clades. Members of the non-bloodfeeding family Haemopidae were scattered throughout the tree and among traditional hirudinid genera. A combination of nuclear 18S rDNA and 28S rDNA with mitochondrial 12S rDNA and cytochrome c oxidase I were analyzed with Parsimony and with Bayesian methods. CONCLUSION: The family Hirudinidae must be refined to include only the clade containing Hirudo medicinalis (European medicinal leech) and related leeches irrespective of bloodfeeding behavior. A second clade containing Macrobdella decora (North American medicinal leech) and its relatives may yet be recognized in Semiscolecidae in order to avoid paraphyly. The African distribution of species from each of the divergent hirudinid clades suggests that a deep divergence took place in the history of the medicinal leeches hundreds of millions of years ago. PMID- 19811661 TI - Refinement of lentiviral vector for improved RNA processing and reduced rates of self inactivation repair. AB - BACKGROUND: Lentiviral gene therapy vectors are now finding clinical application. In order to fully exploit their potential it is important that vectors are made as efficient and as safe as possible. Accordingly, we have modified a previously reported vector to improve RNA processing, minimise Human Immunodeficiency Virus Type-1 (HIV-1) sequence content and reduce repair of the self inactivating (SIN) deletion. RESULTS: HIV-1 sequence in the vector was reduced by substituting the polyadenylation signal with a heterologous signal. Mutation of splice donor sites was undertaken to prevent the majority of splicing within the vector genomic RNA. In addition, a number of other sequences within the vector were deleted. The combination of these modifications was able to significantly reduce the rates of both vector mobilisation and repair of the self inactivating deletion after two rounds of marker rescue. CONCLUSION: RNA processing can be improved by mutation of the major and minor HIV-1 splice donor sites in the vector. In addition the rate of vector mobilisation and repair of SIN vectors can be successfully reduced by careful vector design that reduces homology between the 5' and 3' long terminal repeats (LTRs) to a minimum. PMID- 19811662 TI - Limited copy number-high resolution melting (LCN-HRM) enables the detection and identification by sequencing of low level mutations in cancer biopsies. AB - BACKGROUND: Mutation detection in clinical tumour samples is challenging when the proportion of tumour cells, and thus mutant alleles, is low. The limited sensitivity of conventional sequencing necessitates the adoption of more sensitive approaches. High resolution melting (HRM) is more sensitive than sequencing but identification of the mutation is desirable, particularly when it is important to discriminate false positives due to PCR errors or template degradation from true mutations.We thus developed limited copy number - high resolution melting (LCN-HRM) which applies limiting dilution to HRM. Multiple replicate reactions with a limited number of target sequences per reaction allow low level mutations to be detected. The dilutions used (based on Ct values) are chosen such that mutations, if present, can be detected by the direct sequencing of amplicons with aberrant melting patterns. RESULTS: Using cell lines heterozygous for mutations, we found that the mutations were not readily detected when they comprised 10% of total alleles (20% tumour cells) by sequencing, whereas they were readily detectable at 5% total alleles by standard HRM. LCN-HRM allowed these mutations to be identified by direct sequencing of those positive reactions.LCN-HRM was then used to review formalin-fixed paraffin-embedded (FFPE) clinical samples showing discordant findings between sequencing and HRM for KRAS exon 2 and EGFR exons 19 and 21. Both true mutations present at low levels and sequence changes due to artefacts were detected by LCN-HRM. The use of high fidelity polymerases showed that the majority of the artefacts were derived from the damaged template rather than replication errors during amplification. CONCLUSION: LCN-HRM bridges the sensitivity gap between HRM and sequencing and is effective in distinguishing between artefacts and true mutations. PMID- 19811663 TI - Relevance of MRI in prediction of malignancy of musculoskeletal system--a prospective evaluation. AB - BACKGROUND: The purpose of this study is to evaluate the role of MRI in musculoskeletal tumours, especially, in prediction of malignancy & to compare whether the diagnosis made on MRI correlates with the cytological/histopathological diagnosis. METHODS: 50 consecutive patients presenting in the Outpatient and Inpatient department of the Department of Orthopaedics or Casualty of Jawaharlal Nehru Medical College and Hospital, AMU, Aligarh, India were included in this study. They were subjected to MR examination on 1.5 Tesla superconducting system (MAGNETOM Avanto, Siemens). After localizer sequences, T1W and STIR images were obtained in longitudinal planes followed by T2W and post contrast T1W images in axial planes. Additional sequences were taken when required. Various imaging characteristics of tumours were evaluated statistically and their respective sensitivity and specificity in prediction of malignancy were obtained. RESULTS: Features associated with benign diagnosis in a large percentage of cases, are size less than 8 cm, sharp margination, homogeneous T2 signal, absence of oedema, necrosis, calcification and fluid-fluid levels. Similarly, malignant tumours are commonly associated with presence of irregular margins, inhomogeneous signal intensity, oedema, necrosis, haemorrhage, fascial penetration, bone changes and neurovascular involvement. A correct histological diagnosis is reached on the basis of imaging studies alone in 65% to 75% of cases. The sensitivity for a MRI diagnosis of malignant tumour was 95% and specificity was 84%. CONCLUSION: Differentiation of malignant from benign lesions of musculoskeletal system is best made by a combination of clinical and imaging parameters rather than by any single MR characteristic. When a lesion has a non specific MR imaging appearance, it is useful to formulate a suitably ordered differential diagnosis based on tumour prevalence, patient age, and anatomic location. A systematic approach markedly improves diagnostic results. PMID- 19811664 TI - Low validity of self-report in identifying recent mental health diagnosis among U.S. service members completing Pre-Deployment Health Assessment (PreDHA) and deployed to Afghanistan, 2007: a retrospective cohort study. AB - BACKGROUND: Since 1998, the U.S. Armed Forces has used the mandatory Pre Deployment Health Assessment (PreDHA) screening questionnaire as a means of assessing the health and suitability of U.S. service members for deployment. Limited data exists to quantify the validity of the self-reported PreDHA. This study was conducted to assess the validity of self-reporting in PreDHA to identify deployed service members who have had a recent mental health disorder diagnosis. METHODS: A retrospective cohort study was conducted on 15,195 U.S. service members deployed in support of combat and reconstruction operations in Afghanistan. The Defense Medical Surveillance System (DMSS), the DoD's longitudinal medical surveillance database, was queried to identify cases among the cohort with a recent diagnosis of a pertinent mental health disorder and to obtain those subjects' responses to the PreDHA. RESULTS: Of the study cohort, 11,179 (73.6%) subjects had a PreDHA available within the DMSS at the time of analysis. A total of 615 subjects (4.0%) had one or more mental health disorder diagnoses during the pre-deployment period. Out the 615 subjects with diagnosed mental health disorders, 465 had a PreDHA. Among these, only 224, not quite half, answered in the affirmative to the PreDHA question: "During the past year, have you sought counseling or care for your mental health?" CONCLUSION: This study demonstrates that the self-reported PreDHA has low validity for identifying service members with diagnosed mental health disorders. The development of electronic decision-support systems which automatically screen electronic health records to identify high-risk service members may prove a valuable component of improved pre-deployment screening processes. PMID- 19811665 TI - Anhedonia in schizophrenia and major depression: state or trait? AB - BACKGROUND: In schizophrenia and major depressive disorder, anhedonia (a loss of capacity to feel pleasure) had differently been considered as a premorbid personological trait or as a main symptom of their clinical picture. The aims of this study were to examine the pathological features of anhedonia in schizophrenic and depressed patients, and to investigate its clinical relations with general psychopathology (negative, positive, and depressive dimensions). METHODS: A total of 145 patients (80 schizophrenics and 65 depressed subjects) were assessed using the Physical Anhedonia Scale and the Social Anhedonia Scale (PAS and SAS, respectively), the Scales for the Assessment of Positive and Negative Symptoms (SAPS and SANS, respectively), the Calgary Depression Scale for Schizophrenics (CDSS), and the Hamilton Depression Rating Scale (HDRS). The statistical analysis was performed in two steps. First, the schizophrenic and depressed samples were dichotomised into 'anhedonic' and 'normal hedonic' subgroups (according to the 'double (PAS/SAS) cut-off') and were compared on the general psychopathology scores using the Mann-Whitney Z test. Subsequently, for the total schizophrenic and depressed samples, Spearman correlations were calculated to examine the relation between anhedonia ratings and the other psychopathological parameters. RESULTS: In the schizophrenic sample, anhedonia reached high significant levels only in 45% of patients (n = 36). This 'anhedonic' subgroup was distinguished by high scores in the disorganisation and negative dimensions. Positive correlations of anhedonia with disorganised and negative symptoms were also been detected. In the depressed sample, anhedonia reached high significant levels in only 36.9% of subjects (n = 24). This 'anhedonic' subgroup as distinguished by high scores in the depression severity and negative dimensions. Positive correlations of anhedonia with depressive and negative symptoms were also been detected. CONCLUSION: In the schizophrenic sample, anhedonia seems to be a specific subjective psychopathological experience of the negative and disorganised forms of schizophrenia. In the depressed sample, anhedonia seems to be a specific subjective psychopathological experience of those major depressive disorder forms with a marked clinical depression severity. PMID- 19811667 TI - XVII International AIDS Conference: From Evidence to Action - Introduction. PMID- 19811668 TI - XVII International AIDS Conference: From Evidence to Action - Epidemiology. AB - As the epidemic matures, accurate information about where new infections are occurring, and in which populations, is becoming increasingly critical in designing effective, targeted interventions relevant to current epidemiological trends. Although the quality and accuracy of HIV surveillance data and methodology have improved, in many cases the second generation WHO/UNAIDS surveillance system has not been fully implemented at the national level. National surveillance systems in many low and middle-income countries often do not collect disaggregated data on some most at risk populations, which is critical to developing targeted prevention interventions.While the majority of new infections occur in low- and middle-income countries, the dynamic situation in high-income countries demands renewed attention. PMID- 19811669 TI - XVII International AIDS Conference: From Evidence to Action - Basic science. AB - This article focuses on the sessions in which basic science research was presented at the XVII International AIDS Conference (AIDS 2008). It also provides an analysis of basic science issues which generated significant discussion and debate at the conference and are likely to have implications for future laboratory and clinical research. Data presented at AIDS 2008 confirmed the speed with which HIV establishes latent viral reservoirs following infection and the resulting challenges to viral eradication given how effectively HIV proviral RNA inserts itself into human DNA within these reservoirs. Studies also raised questions about the source of residual viremia and how these might be targeted by novel therapeutic approaches. PMID- 19811666 TI - Consanguinity and reproductive health among Arabs. AB - Consanguineous marriages have been practiced since the early existence of modern humans. Until now consanguinity is widely practiced in several global communities with variable rates depending on religion, culture, and geography. Arab populations have a long tradition of consanguinity due to socio-cultural factors. Many Arab countries display some of the highest rates of consanguineous marriages in the world, and specifically first cousin marriages which may reach 25-30% of all marriages. In some countries like Qatar, Yemen, and UAE, consanguinity rates are increasing in the current generation. Research among Arabs and worldwide has indicated that consanguinity could have an effect on some reproductive health parameters such as postnatal mortality and rates of congenital malformations. The association of consanguinity with other reproductive health parameters, such as fertility and fetal wastage, is controversial. The main impact of consanguinity, however, is an increase in the rate of homozygotes for autosomal recessive genetic disorders. Worldwide, known dominant disorders are more numerous than known recessive disorders. However, data on genetic disorders in Arab populations as extracted from the Catalogue of Transmission Genetics in Arabs (CTGA) database indicate a relative abundance of recessive disorders in the region that is clearly associated with the practice of consanguinity. PMID- 19811670 TI - XVII International AIDS Conference: From Evidence to Action - Clinical and biomedical prevention science. AB - The question of whether to initiate ART at higher CD4+ cell counts than currently recommended by World Health Organization (WHO) treatment guidelines received much attention at the XVII International AIDS Conference (AIDS 2008). If studies presented at the conference ultimately lead to a revision of WHO treatment guidance, the estimated number of people who will need ART globally will increase substantially. Task-shifting is emerging as an important strategy for dealing with the acute shortage of health care workers in many high-burden countries, and several studies presented at AIDS 2008 demonstrated the impressive health system efficiencies garnered by using nurses or other health care providers to deliver HIV care and treatment. Other key presentations and discussion at the conference focused on the optimal time to start TB treatment in HIV-infected patients, the growing risk of resistance in high-burden countries, including its impact on future treatment options, and several large cohort trials testing optimal drug regimens in resource-limited settings.Biomedical prevention research continues to confirm the long-term, protective benefits of circumcision. Several studies involving HIV serodiscordant heterosexual couples have produced data suggesting a strong protective effect of ART for HIV-negative partners. Disappointing results from recent vaccine and non-ARV based microbicides trials are nevertheless providing important data to this field, and the expanding number of pre-exposure prophylaxis (PrEP) trials and ARV-based microbicides appear to provide the best hope for a new, efficacious biomedical prevention intervention. PMID- 19811671 TI - XVII International AIDS Conference: From Evidence to Action - Social, behavioural and economic science and policy and political science. AB - AIDS 2008 firmly established stigma and discrimination as fundamental priorities in the push for universal access to HIV prevention, treatment, care and support. Conference sessions and discussions reinforced the tangible negative effects of stigma on national legislation and policies. A strong theme throughout the conference was the need to replace prevention interventions that focus exclusively on individual behaviour change or biomedical prevention interventions with "combination prevention" approaches that address both individual and structural factors that increase vulnerability to HIV infection.Several high level sessions addressed various aspects of the debate over "vertical" (disease specific) versus "horizontal" (health systems) funding. The majority of evidence presented at the conference suggests that HIV investments strengthen health systems through the establishment of clinical and laboratory infrastructure, strengthened supply and procurement systems, improvements in health care worker training, and increased community engagement.Human rights were a focal point at the conference; several presentations emphasized the importance of securing human rights to achieve universal access goals, including workplace discrimination, travel restrictions, gender inequality, and the criminalization of homosexuality, drug use, sex work, and HIV transmission and/or exposure. PMID- 19811672 TI - XVII International AIDS Conference: From Evidence to Action - Regional focus. AB - This article summarizes the challenges, opportunities and lessons learned from presentations, discussions and debates addressing major policy and programmatic responses to HIV in six geographical regions: Sub-Saharan Africa, Asia and the Pacific, Eastern Europe and Central Asia, Latin America, Caribbean, and Middle East and North Africa. It draws from AIDS 2008 Leadership and Community Programmes, particularly the six regional sessions, and Global Village activities.While the epidemiological, cultural and socio-economic contexts in these regions vary considerably, several common, overarching principles and themes emerged. They include: advancing basic human rights, particularly for vulnerable and most at risk populations; ensuring the sustainability of the HIV response through long-term, predictable financing; strengthening health systems; investing in strategic health information; and improving accountability and the involvement of civil society in the response to AIDS.Equally important is the need to address political barriers to implementing evidence-based interventions such as opioid substitution therapy (OST), needle and syringe programmes (NSPs), comprehensive sexuality education for youth, and sexual and reproductive rights. Finally, these regional discussions emphasized the need for legislative and policy reforms related to structural barriers facing women and girls, MSM, IDUs, sex workers and migrant populations. PMID- 19811673 TI - XVII International AIDS Conference: From Evidence to Action - AIDS 2008 and the global response to AIDS. AB - The impact of the XVII International AIDS Conference (AIDS 2008) was reflected in a number of commitments from political and business leaders, who announced initiatives ranging from implementing comprehensive sexual education for young people in Latin America to reducing regulatory barriers and the price of drugs in the host country. The unprecedented media coverage brought attention and public awareness to the epidemic in Latin America.Several meetings and sessions at AIDS 2008 also addressed the potential for the International AIDS Conference to play an even stronger role in tracking progress towards universal access and in improving accountability in the global response to AIDS, particularly given some of the inherent weaknesses in the United Nations General Assembly Special Session (UNGASS) review process. The impact of AIDS 2008 was strongest in Mexico, the host country, and in Latin America. Highlights included the policy changes announced by President Calderon on pharmaceutical manufacturing to the focus on sex workers and gay and other MSM in marches, activism and the conference programme.The next two years will determine whether the successes reported in Mexico are sustained and whether there is progress in addressing the barriers that continue to hamper an evidence-based response to HIV/AIDS. The next International AIDS Conference is scheduled for the universal address deadline of 2010. PMID- 19811675 TI - Analysis and modeling of time-course gene-expression profiles from nanomaterial exposed primary human epidermal keratinocytes. AB - BACKGROUND: Nanomaterials are being manufactured on a commercial scale for use in medical, diagnostic, energy, component and communications industries. However, concerns over the safety of engineered nanomaterials have surfaced. Humans can be exposed to nanomaterials in different ways such as inhalation or exposure through the integumentary system. RESULTS: The interactions of engineered nanomaterials with primary human cells was investigated, using a systems biology approach combining gene expression microarray profiling with dynamic experimental parameters. In this experiment, primary human epidermal keratinocytes cells were exposed to several low-micron to nano-scale materials, and gene expression was profiled over both time and dose to compile a comprehensive picture of nanomaterial-cellular interactions. Very few gene-expression studies so far have dealt with both time and dose response simultaneously. Here, we propose different approaches to this kind of analysis. First, we used heat maps and multi dimensional scaling (MDS) plots to visualize the dose response of nanomaterials over time. Then, in order to find out the most common patterns in gene-expression profiles, we used self-organizing maps (SOM) combined with two different criteria to determine the number of clusters. The consistency of SOM results is discussed in context of the information derived from the MDS plots. Finally, in order to identify the genes that have significantly different responses among different levels of dose of each treatment while accounting for the effect of time at the same time, we used a two-way ANOVA model, in connection with Tukey's additivity test and the Box-Cox transformation. The results are discussed in the context of the cellular responses of engineered nanomaterials. CONCLUSION: The analysis presented here lead to interesting and complementary conclusions about the response across time of human epidermal keratinocytes after exposure to nanomaterials. For example, we observed that gene expression for most treatments become closer to the expression of the baseline cultures as time proceeds. The genes found to be differentially-expressed are involved in a number of cellular processes, including regulation of transcription and translation, protein localization, transport, cell cycle progression, cell migration, cytoskeletal reorganization, signal transduction, and development. PMID- 19811674 TI - Proceedings of the 2009 MidSouth Computational Biology and Bioinformatics Society (MCBIOS) conference. Introduction. PMID- 19811676 TI - Site-specific impacts on gene expression and behavior in fathead minnows (Pimephales promelas) exposed in situ to streams adjacent to sewage treatment plants. AB - BACKGROUND: Environmental monitoring for pharmaceuticals and endocrine disruptors in the aquatic environment traditionally employs a variety of methods including analytical chemistry, as well as a variety of histological and biochemical endpoints that correlate with the fish fitness. It is now clear that analytical chemistry alone is insufficient to identify aquatic environments that are compromised because these measurements do not identify the biologically available dose. The biological endpoints that are measured are important because they relate to known impairments; however, they are not specific to the contaminants and often focus on only a few known endpoints. These studies can be enhanced by looking more broadly at changes in gene expression, especially if the analysis focuses on biochemical pathways. The present study was designed to obtain additional information for well-characterized sites adjacent to sewage treatment plants in MN that are thought to be impacted by endocrine disruptors. RESULTS: Here we examine five sites that have been previously characterized and examine changes in gene expression in fathead minnows (Pimephales promelas) that have been caged for 48 h in each of the aquatic environments. We find that the gene expression changes are characteristic and unique at each of the five sites. Also, fish exposed to two of the sites, 7 and 12, present a more aggressive behavior compared to control fish. CONCLUSION: Our results show that a short-term exposure to sewage treatment plant effluents was able to induce a site-specific gene expression pattern in the fathead minnow gonad and liver. The short-term exposure was also enough to affect fish sexual behavior. Our results also show that microarray analysis can be very useful at determining potential exposure to chemicals, and could be used routinely as a tool for environmental monitoring. PMID- 19811677 TI - Microarray platform consistency is revealed by biologically functional analysis of gene expression profiles. AB - BACKGROUND: Several different microarray platforms are available for measuring gene expression. There are disagreements within the microarray scientific community for intra- and inter-platform consistency of these platforms. Both high and low consistencies were demonstrated across different platforms in terms of genes with significantly differential expression. Array studies for gene expression are used to explore biological causes and effects. Therefore, consistency should eventually be evaluated in a biological setting to reveal the functional differences between the examined samples, not just a list of differentially expressed genes (DEG). In this study, we investigated whether different platforms had a high consistency from the biologically functional perspective. RESULTS: DEG data without filtering the different probes in microarrays from different platforms generated from kidney samples of rats treated with the kidney carcinogen, aristolochic acid, in five test sites using microarrays from Affymetrix, Applied Biosystems, Agilent, and GE health platforms (two sites using Affymetrix for intra-platform comparison) were input into the Ingenuity Pathway Analysis (IPA) system for functional analysis. The functions of the DEG lists determined by IPA were compared across the four different platforms and two test sites for Affymetrix platform. Analysis results showed that there is a very high level of consistency between the two test sites using the same platform or among different platforms. The top functions determined by the different platforms were very similar and reflected carcinogenicity and toxicity of aristolochic acid in the rat kidney. CONCLUSION: Our results demonstrate that highly consistent biological information can be generated from different microarray platforms. PMID- 19811678 TI - Automatic identification of angiogenesis in double stained images of liver tissue. AB - BACKGROUND: To grow beyond certain size and reach oxygen and other essential nutrients, solid tumors trigger angiogenesis (neovascularization) by secreting various growth factors. Based on this fact, several researches proposed that density of newly formed vessels correlate with tumor malignancy. Vessel density is known as a true prognostic indicator for several types of cancer. However, automated quantification of angiogenesis is still in its primitive stage, and deserves more intelligent methods by taking advantages accruing from novel computer algorithms. RESULTS: The newly introduced characteristics of subimages performed well in identification of region-of-angiogenesis. The proposed technique was tested on 522 samples collected from two high-resolution tissues. Having 0.90 overall f-measure, the results obtained with Support Vector Machines show significant agreement between automated framework and manual assessment of microvessels. CONCLUSION: This study introduces a new framework to identify angiogenesis to measure microvessel density (MVD) in digitalized images of liver cancer tissues. The objective is to recognize all subimages having new vessel formations. In addition to region based characteristics, a set of morphological features are proposed to differentiate positive and negative incidences. PMID- 19811679 TI - NATbox: a network analysis toolbox in R. AB - BACKGROUND: There has been recent interest in capturing the functional relationships (FRs) from high-throughput assays using suitable computational techniques. FRs elucidate the working of genes in concert as a system as opposed to independent entities hence may provide preliminary insights into biological pathways and signalling mechanisms. Bayesian structure learning (BSL) techniques and its extensions have been used successfully for modelling FRs from expression profiles. Such techniques are especially useful in discovering undocumented FRs, investigating non-canonical signalling mechanisms and cross-talk between pathways. The objective of the present study is to develop a graphical user interface (GUI), NATbox: Network Analysis Toolbox in the language R that houses a battery of BSL algorithms in conjunction with suitable statistical tools for modelling FRs in the form of acyclic networks from gene expression profiles and their subsequent analysis. RESULTS: NATbox is a menu-driven open-source GUI implemented in the R statistical language for modelling and analysis of FRs from gene expression profiles. It provides options to (i) impute missing observations in the given data (ii) model FRs and network structure from gene expression profiles using a battery of BSL algorithms and identify robust dependencies using a bootstrap procedure, (iii) present the FRs in the form of acyclic graphs for visualization and investigate its topological properties using network analysis metrics, (iv) retrieve FRs of interest from published literature. Subsequently, use these FRs as structural priors in BSL (v) enhance scalability of BSL across high-dimensional data by parallelizing the bootstrap routines. CONCLUSION: NATbox provides a menu-driven GUI for modelling and analysis of FRs from gene expression profiles. By incorporating readily available functions from existing R-packages, it minimizes redundancy and improves reproducibility, transparency and sustainability, characteristic of open-source environments. NATbox is especially suited for interdisciplinary researchers and biologists with minimal programming experience and would like to use systems biology approaches without delving into the algorithmic aspects. The GUI provides appropriate parameter recommendations for the various menu options including default parameter choices for the user. NATbox can also prove to be a useful demonstration and teaching tool in graduate and undergraduate course in systems biology. It has been tested successfully under Windows and Linux operating systems. The source code along with installation instructions and accompanying tutorial can be found at http://bioinformatics.ualr.edu/natboxWiki/index.php/Main_Page. PMID- 19811680 TI - Protein local 3D structure prediction by Super Granule Support Vector Machines (Super GSVM). AB - BACKGROUND: Understanding the relationship between the protein sequence and the 3D structure is a major research area in bioinformatics. The prediction of complete protein tertiary structure based only on sequence information is still an impractical work. This paper aims at revealing the hidden knowledge of the sequence motifs and the local tertiary structure. RESULTS: In this paper, we propose a Super Granule Support Vector Machine (Super GSVM) model to obtain the high quality protein sequence motifs and to predict local tertiary structure information based on purely sequence information. CONCLUSION: The proposed model overcomes the innate shortcoming of using the SVM on such a large data set, which is the inherent computational complexity involved in training support vectors for huge datasets including half million of samples. The satisfactory prediction results show the Super GSVM model generates decent protein sequence clusters and has the ability to capture the hidden sequence-to-structure information. This model also has a strong potential in the application of SVMs on other research areas with huge datasets. PMID- 19811681 TI - Novel software package for cross-platform transcriptome analysis (CPTRA). AB - BACKGROUND: Next-generation sequencing techniques enable several novel transcriptome profiling approaches. Recent studies indicated that digital gene expression profiling based on short sequence tags has superior performance as compared to other transcriptome analysis platforms including microarrays. However, the transcriptomic analysis with tag-based methods often depends on available genome sequence. The use of tag-based methods in species without genome sequence should be complemented by other methods such as cDNA library sequencing. The combination of different next generation sequencing techniques like 454 pyrosequencing and Illumina Genome Analyzer (Solexa) will enable high-throughput and accurate global gene expression profiling in species with limited genome information. The combination of transcriptome data acquisition methods requires cross-platform transcriptome data analysis platforms, including a new software package for data processing. RESULTS: Here we presented a software package, CPTRA: Cross-Platform TRanscriptome Analysis, to analyze transcriptome profiling data from separate methods. The software package is available at http://people.tamu.edu/approximately syuan/cptra/cptra.html. It was applied to the case study of non-target site glyphosate resistance in horseweed; and the data was mined to discover resistance target gene(s). For the software, the input data included a long-read sequence dataset with proper annotation, and a short read sequence tag dataset for the quantification of transcripts. By combining the two datasets, the software carries out the unique sequence tag identification, tag counting for transcript quantification, and cross-platform sequence matching functions, whereby the short sequence tags can be annotated with a function, level of expression, and Gene Ontology (GO) classification. Multiple sequence search algorithms were implemented and compared. The analysis highlighted the importance of transport genes in glyphosate resistance and identified several candidate genes for down-stream analysis. CONCLUSION: CPTRA is a powerful software package for next generation sequencing-based transcriptome profiling in species with limited genome information. According to our case study, the strategy can greatly broaden the application of the next generation sequencing for transcriptome analysis in species without reference genome sequence. PMID- 19811682 TI - An automated proteomic data analysis workflow for mass spectrometry. AB - BACKGROUND: Mass spectrometry-based protein identification methods are fundamental to proteomics. Biological experiments are usually performed in replicates and proteomic analyses generate huge datasets which need to be integrated and quantitatively analyzed. The Sequest search algorithm is a commonly used algorithm for identifying peptides and proteins from two dimensional liquid chromatography electrospray ionization tandem mass spectrometry (2-D LC ESI MS(2)) data. A number of proteomic pipelines that facilitate high throughput 'post data acquisition analysis' are described in the literature. However, these pipelines need to be updated to accommodate the rapidly evolving data analysis methods. Here, we describe a proteomic data analysis pipeline that specifically addresses two main issues pertinent to protein identification and differential expression analysis: 1) estimation of the probability of peptide and protein identifications and 2) non-parametric statistics for protein differential expression analysis. Our proteomic analysis workflow analyzes replicate datasets from a single experimental paradigm to generate a list of identified proteins with their probabilities and significant changes in protein expression using parametric and non-parametric statistics. RESULTS: The input for our workflow is Bioworks 3.2 Sequest (or a later version, including cluster) output in XML format. We use a decoy database approach to assign probability to peptide identifications. The user has the option to select "quality thresholds" on peptide identifications based on the P value. We also estimate probability for protein identification. Proteins identified with peptides at a user-specified threshold value from biological experiments are grouped as either control or treatment for further analysis in ProtQuant. ProtQuant utilizes a parametric (ANOVA) method, for calculating differences in protein expression based on the quantitative measure SigmaXcorr. Alternatively ProtQuant output can be further processed using non-parametric Monte-Carlo resampling statistics to calculate P values for differential expression. Correction for multiple testing of ANOVA and resampling P values is done using Benjamini and Hochberg's method. The results of these statistical analyses are then combined into a single output file containing a comprehensive protein list with probabilities and differential expression analysis, associated P values, and resampling statistics. CONCLUSION: For biologists carrying out proteomics by mass spectrometry, our workflow facilitates automated, easy to use analyses of Bioworks (3.2 or later versions) data. All the methods used in the workflow are peer-reviewed and as such the results of our workflow are compliant with proteomic data submission guidelines to public proteomic data repositories including PRIDE. Our workflow is a necessary intermediate step that is required to link proteomics data to biological knowledge for generating testable hypotheses. PMID- 19811683 TI - PathBinder--text empirics and automatic extraction of biomolecular interactions. AB - MOTIVATION: The increasingly large amount of free, online biological text makes automatic interaction extraction correspondingly attractive. Machine learning is one strategy that works by uncovering and using useful properties that are implicit in the text. However these properties are usually not reported in the literature explicitly. By investigating specific properties of biological text passages in this paper, we aim to facilitate an alternative strategy, the use of text empirics, to support mining of biomedical texts for biomolecular interactions. We report on our application of this approach, and also report some empirical findings about an important class of passages. These may be useful to others who may also wish to use the empirical properties we describe. RESULTS: We manually analyzed syntactic and semantic properties of sentences likely to describe interactions between biomolecules. The resulting empirical data were used to design an algorithm for the PathBinder system to extract biomolecular interactions from texts. PathBinder searches PubMed for sentences describing interactions between two given biomolecules. PathBinder then uses probabilistic methods to combine evidence from multiple relevant sentences in PubMed to assess the relative likelihood of interaction between two arbitrary biomolecules. A biomolecular interaction network was constructed based on those likelihoods. CONCLUSION: The text empirics approach used here supports computationally friendly, performance competitive, automatic extraction of biomolecular interactions from texts. AVAILABILITY: http://www.metnetdb.org/pathbinder. PMID- 19811684 TI - Graph ranking for exploratory gene data analysis. AB - BACKGROUND: Microarray technology has made it possible to simultaneously monitor the expression levels of thousands of genes in a single experiment. However, the large number of genes greatly increases the challenges of analyzing, comprehending and interpreting the resulting mass of data. Selecting a subset of important genes is inevitable to address the challenge. Gene selection has been investigated extensively over the last decade. Most selection procedures, however, are not sufficient for accurate inference of underlying biology, because biological significance does not necessarily have to be statistically significant. Additional biological knowledge needs to be integrated into the gene selection procedure. RESULTS: We propose a general framework for gene ranking. We construct a bipartite graph from the Gene Ontology (GO) and gene expression data. The graph describes the relationship between genes and their associated molecular functions. Under a species condition, edge weights of the graph are assigned to be gene expression level. Such a graph provides a mathematical means to represent both species-independent and species-dependent biological information. We also develop a new ranking algorithm to analyze the weighted graph via a kernelized spatial depth (KSD) approach. Consequently, the importance of gene and molecular function can be simultaneously ranked by a real-valued measure, KSD, which incorporates the global and local structure of the graph. Over-expressed and under-regulated genes also can be separately ranked. CONCLUSION: The gene function bigraph integrates molecular function annotations into gene expression data. The relevance of genes is described in the graph (through a common function). The proposed method provides an exploratory framework for gene data analysis. PMID- 19811685 TI - Facilitating functional annotation of chicken microarray data. AB - BACKGROUND: Modeling results from chicken microarray studies is challenging for researchers due to little functional annotation associated with these arrays. The Affymetrix GenChip chicken genome array, one of the biggest arrays that serve as a key research tool for the study of chicken functional genomics, is among the few arrays that link gene products to Gene Ontology (GO). However the GO annotation data presented by Affymetrix is incomplete, for example, they do not show references linked to manually annotated functions. In addition, there is no tool that facilitates microarray researchers to directly retrieve functional annotations for their datasets from the annotated arrays. This costs researchers amount of time in searching multiple GO databases for functional information. RESULTS: We have improved the breadth of functional annotations of the gene products associated with probesets on the Affymetrix chicken genome array by 45% and the quality of annotation by 14%. We have also identified the most significant diseases and disorders, different types of genes, and known drug targets represented on Affymetrix chicken genome array. To facilitate functional annotation of other arrays and microarray experimental datasets we developed an Array GO Mapper (AGOM) tool to help researchers to quickly retrieve corresponding functional information for their dataset. CONCLUSION: Results from this study will directly facilitate annotation of other chicken arrays and microarray experimental datasets. Researchers will be able to quickly model their microarray dataset into more reliable biological functional information by using AGOM tool. The disease, disorders, gene types and drug targets revealed in the study will allow researchers to learn more about how genes function in complex biological systems and may lead to new drug discovery and development of therapies. The GO annotation data generated will be available for public use via AgBase website and will be updated on regular basis. PMID- 19811686 TI - Integrating phenotype and gene expression data for predicting gene function. AB - BACKGROUND: This paper presents a framework for integrating disparate data sets to predict gene function. The algorithm constructs a graph, called an integrated similarity graph, by computing similarities based upon both gene expression and textual phenotype data. This integrated graph is then used to make predictions about whether individual genes should be assigned a particular annotation from the Gene Ontology. RESULTS: A combined graph was generated from publicly available gene expression data and phenotypic information from Saccharomyces cerevisiae. This graph was used to assign annotations to genes, as were graphs constructed from gene expression data and textual phenotype information alone. While the F-measure appeared similar for all three methods, annotations based upon the integrated similarity graph exhibited a better overall precision than gene expression or phenotype information alone can generate. The integrated approach was also able to assign almost as many annotations as the gene expression method alone, and generated significantly more total and correct assignments than the phenotype information could provide. CONCLUSION: These results suggest that augmenting standard gene expression data sets with publicly available textual phenotype data can help generate more precise functional annotation predictions while mitigating the weaknesses of a standard textual phenotype approach. PMID- 19811688 TI - Threshold selection in gene co-expression networks using spectral graph theory techniques. AB - BACKGROUND: Gene co-expression networks are often constructed by computing some measure of similarity between expression levels of gene transcripts and subsequently applying a high-pass filter to remove all but the most likely biologically-significant relationships. The selection of this expression threshold necessarily has a significant effect on any conclusions derived from the resulting network. Many approaches have been taken to choose an appropriate threshold, among them computing levels of statistical significance, accepting only the top one percent of relationships, and selecting an arbitrary expression cutoff. RESULTS: We apply spectral graph theory methods to develop a systematic method for threshold selection. Eigenvalues and eigenvectors are computed for a transformation of the adjacency matrix of the network constructed at various threshold values. From these, we use a basic spectral clustering method to examine the set of gene-gene relationships and select a threshold dependent upon the community structure of the data. This approach is applied to two well-studied microarray data sets from Homo sapiens and Saccharomyces cerevisiae. CONCLUSION: This method presents a systematic, data-based alternative to using more artificial cutoff values and results in a more conservative approach to threshold selection than some other popular techniques such as retaining only statistically significant relationships or setting a cutoff to include a percentage of the highest correlations. PMID- 19811687 TI - Comparative genome analysis of lignin biosynthesis gene families across the plant kingdom. AB - BACKGROUND: As a major component of plant cell wall, lignin plays important roles in mechanical support, water transport, and stress responses. As the main cause for the recalcitrance of plant cell wall, lignin modification has been a major task for bioenergy feedstock improvement. The study of the evolution and function of lignin biosynthesis genes thus has two-fold implications. First, the lignin biosynthesis pathway provides an excellent model to study the coordinative evolution of a biochemical pathway in plants. Second, understanding the function and evolution of lignin biosynthesis genes will guide us to develop better strategies for bioenergy feedstock improvement. RESULTS: We analyzed lignin biosynthesis genes from fourteen plant species and one symbiotic fungal species. Comprehensive comparative genome analysis was carried out to study the distribution, relatedness, and family expansion of the lignin biosynthesis genes across the plant kingdom. In addition, we also analyzed the comparative synteny map between rice and sorghum to study the evolution of lignin biosynthesis genes within the Poaceae family and the chromosome evolution between the two species. Comprehensive lignin biosynthesis gene expression analysis was performed in rice, poplar and Arabidopsis. The representative data from rice indicates that different fates of gene duplications exist for lignin biosynthesis genes. In addition, we also carried out the biomass composition analysis of nine Arabidopsis mutants with both MBMS analysis and traditional wet chemistry methods. The results were analyzed together with the genomics analysis. CONCLUSION: The research revealed that, among the species analyzed, the complete lignin biosynthesis pathway first appeared in moss; the pathway is absent in green algae. The expansion of lignin biosynthesis gene families correlates with substrate diversity. In addition, we found that the expansion of the gene families mostly occurred after the divergence of monocots and dicots, with the exception of the C4H gene family. Gene expression analysis revealed different fates of gene duplications, largely confirming plants are tolerant to gene dosage effects. The rapid expansion of lignin biosynthesis genes indicated that the translation of transgenic lignin modification strategies from model species to bioenergy feedstock might only be successful between the closely relevant species within the same family. PMID- 19811689 TI - HPD: an online integrated human pathway database enabling systems biology studies. AB - BACKGROUND: Pathway-oriented experimental and computational studies have led to a significant accumulation of biological knowledge concerning three major types of biological pathway events: molecular signaling events, gene regulation events, and metabolic reaction events. A pathway consists of a series of molecular pathway events that link molecular entities such as proteins, genes, and metabolites. There are approximately 300 biological pathway resources as of April 2009 according to the Pathguide database; however, these pathway databases generally have poor coverage or poor quality, and are difficult to integrate, due to syntactic-level and semantic-level data incompatibilities. RESULTS: We developed the Human Pathway Database (HPD) by integrating heterogeneous human pathway data that are either curated at the NCI Pathway Interaction Database (PID), Reactome, BioCarta, KEGG or indexed from the Protein Lounge Web sites. Integration of pathway data at syntactic, semantic, and schematic levels was based on a unified pathway data model and data warehousing-based integration techniques. HPD provides a comprehensive online view that connects human proteins, genes, RNA transcripts, enzymes, signaling events, metabolic reaction events, and gene regulatory events. At the time of this writing HPD includes 999 human pathways and more than 59,341 human molecular entities. The HPD software provides both a user-friendly Web interface for online use and a robust relational database backend for advanced pathway querying. This pathway tool enables users to 1) search for human pathways from different resources by simply entering genes/proteins involved in pathways or words appearing in pathway names, 2) analyze pathway-protein association, 3) study pathway-pathway similarity, and 4) build integrated pathway networks. We demonstrated the usage and characteristics of the new HPD through three breast cancer case studies. CONCLUSION: HPD http://bio.informatics.iupui.edu/HPD is a new resource for searching, managing, and studying human biological pathways. Users of HPD can search against large collections of human biological pathways, compare related pathways and their molecular entity compositions, and build high-quality, expanded-scope disease pathway models. The current HPD software can help users address a wide range of pathway-related questions in human disease biology studies. PMID- 19811690 TI - Computational analysis of gene expression space associated with metastatic cancer. AB - BACKGROUND: Prostate carcinoma is among the most common types of cancer affecting hundreds of thousands people every year. Once the metastatic form of prostate carcinoma is documented, the majority of patients die from their tumors as opposed to other causes. The key to successful treatment is in the earliest possible diagnosis, as well as understanding the molecular mechanisms of metastatic progression. A number of recent studies have identified multiple biomarkers for metastatic progression. However, most of the studies consider only direct comparison between metastatic and non-metastatic classes of samples. RESULTS: We propose an alternative concept of analysis that considers the entire multidimensional space of gene expression and identifies the partition of this space in which metastatic development is possible. To apply this concept in cancer gene expression studies we utilize a modification of high-dimension natural taxonomy algorithm FOREL. Our analysis of microarray data containing primary and metastatic cancer samples has revealed not only differentially expressed genes, but also relations between different groups of primary and metastatic cancer. Metastatic samples tend to occupy a distinct partition of gene expression space. Further pathway analysis suggests that this partition is delineated by a specific pattern of gene expression in cytoskeleton remodeling, cell adhesion and apoptosis/cell survival pathways. We compare our findings with both report of original analysis and recent studies in molecular mechanism of metastasis. CONCLUSION: Our analysis indicates a single molecular mechanism of metastasis. The new approach does not contradict previously reported findings, but reveals important details unattainable with traditional methodology. PMID- 19811691 TI - Exploratory visual analysis of conserved domains on multiple sequence alignments. AB - BACKGROUND: Multiple alignment of protein sequences can provide insight into sequence conservation across many species and thus allow identification of those sections of the sequence most critical to protein function. This insight can be augmented by joint display of conserved domains along the sequences. By fusing this metadata visually, biologists can analyze sequence conservation and functional motifs simultaneously and efficiently. RESULTS: We present MSAVis, a new approach combining luminance and hue for simultaneous visualization of conserved motifs and sequence alignment. Input for the algorithm is a multiple sequence alignment in a standard format. The NCBI Conserved Domain Database (CDD) is used for finding conserved domains along the alignment. The visualization quickly identifies conserved domains, and allows both macro (sequence-long) and micro (small amino-acid neighborhood) views. CONCLUSION: MSAVis utilizes two visual cues, luminance and hue, to facilitate at-a-glance summary of the conservation of a user-provided protein alignment while enabling multiple comparisons among functional domains. These visual cues are preattentive and separable so that the relationship between conservation strength and domain membership can be understood. The MSAVis software, written in Python and using BioPython and OpenGL, can be found at http://agbase.msstate.edu/tools/MSAVis.html. PMID- 19811692 TI - Structural and functional-annotation of an equine whole genome oligoarray. AB - BACKGROUND: The horse genome is sequenced, allowing equine researchers to use high-throughput functional genomics platforms such as microarrays; next generation sequencing for gene expression and proteomics. However, for researchers to derive value from these functional genomics datasets, they must be able to model this data in biologically relevant ways; to do so requires that the equine genome be more fully annotated. There are two interrelated types of genomic annotation: structural and functional. Structural annotation is delineating and demarcating the genomic elements (such as genes, promoters, and regulatory elements). Functional annotation is assigning function to structural elements. The Gene Ontology (GO) is the de facto standard for functional annotation, and is routinely used as a basis for modelling and hypothesis testing, large functional genomics datasets. RESULTS: An Equine Whole Genome Oligonucleotide (EWGO) array with 21,351 elements was developed at Texas A&M University. This 70-mer oligoarray was designed using the approximately 7 x assembled and annotated sequence of the equine genome to be one of the most comprehensive arrays available for expressed equine sequences. To assist researchers in determining the biological meaning of data derived from this array, we have structurally annotated it by mapping the elements to multiple database accessions, including UniProtKB, Entrez Gene, NRPD (Non-Redundant Protein Database) and UniGene. We next provided GO functional annotations for the gene transcripts represented on this array. Overall, we GO annotated 14,531 gene products (68.1% of the gene products represented on the EWGO array) with 57,912 annotations. GAQ (GO Annotation Quality) scores were calculated for this array both before and after we added GO annotation. The additional annotations improved the meanGAQ score 16-fold. This data is publicly available at AgBase http://www.agbase.msstate.edu/. CONCLUSION: Providing additional information about the public databases which link to the gene products represented on the array allows users more flexibility when using gene expression modelling and hypothesis-testing computational tools. Moreover, since different databases provide different types of information, users have access to multiple data sources. In addition, our GO annotation underpins functional modelling for most gene expression analysis tools and enables equine researchers to model large lists of differentially expressed transcripts in biologically relevant ways. PMID- 19811693 TI - Comparing gene annotation enrichment tools for functional modeling of agricultural microarray data. AB - The widespread availability of microarray technology has driven functional genomics to the forefront as scientists seek to draw meaningful biological conclusions from their microarray results. Gene annotation enrichment analysis is a functional analysis technique that has gained widespread attention and for which many tools have been developed. Unfortunately, most of these tools have limited support for agricultural species. Here, we evaluate and compare four publicly available computational tools (Onto-Express, EasyGO, GOstat, and DAVID) that support analysis of gene expression datasets in agricultural species. We use AgBase as the functional annotation reference for agricultural species. The selected tools were evaluated based on i) available features, usage and accessibility, ii) implemented statistical computational methods, and iii) annotation and enrichment performance analysis. Annotation was assessed using a randomly selected test gene annotation set and an experimental differentially expressed gene-set--both from chicken. The experimental set was also used to evaluate identification of enriched functional groups.Comparison of the tools shows that they produce different sets of annotations for the two datasets and different functional groups for the experimental dataset. While DAVID, GOstat and Onto-Express annotate comparable numbers of genes, DAVID provides by far the most annotations per gene. However, many of DAVID's annotations appear to be redundant or are at very high levels in the GO hierarchy. The GOSlim distribution of annotations shows that GOstat, Onto-Express and EasyGO provide similar GO distributions to those found in AgBase while annotations from DAVID show a different GOSlim distribution, again probably due to duplication and many non specific terms. No consistent trends were found in results of GO term over/under representation analysis applied to the experimental data using different tools. While GOstat, David and Onto-Express could retrieve some significantly enriched terms, EasyGO did not show any significantly enriched terms. There was little agreement about the enriched terms identified by the tools. CONCLUSION: Different tools for functionally annotating gene sets and identifying significantly enriched GO categories differ widely in their results when applied to a test annotation gene set and an experimental dataset from chicken. These results emphasize the need for care when interpreting the results of such analysis and the lack of standardization of approaches. PMID- 19811694 TI - The red wine polyphenol resveratrol reduces polycyclic aromatic hydrocarbon induced DNA damage in MCF-10A cells. AB - Polycyclic aromatic hydrocarbons (PAH) are procarcinogens that can be commonly found in our food and environment. Upon biotransformation in our body system, they can cause DNA damage through the generation of genotoxic species and oxidative stress. Phase I and II enzymes are pivotal in the process of proximate carcinogen formation and elimination. Some dietary phytochemicals are strong inhibitors to the phase I enzymes. In the present study, we investigated the effect of the red wine compound resveratrol on DNA damage induced by PAH in a non tumorigenic breast cell line MCF-10A. Resveratrol ranging from 1 to 5 microm could significantly suppress the expressions of cytochrome P450 (CYP) 1A1, CYP1B1 and UDP-glucuronosyltransferase (UGT) 1A1 induced by 7,12 dimethylbenz[a]anthracene (DMBA). The comet assay indicated that DMBA introduced DNA damage to these cells, and co-treatment of resveratrol at 5 or 10 microm could alleviate the damage. Further investigation illustrated that resveratrol reduced the binding of DMBA metabolites to DNA with no effect on DMBA-induced oxidative DNA damage. Since the phase II enzyme UGT1A1 was suppressed, the elimination of DMBA metabolites would not have contributed to the reduction in the DMBA metabolite-DNA binding. In summary, resveratrol might protect breast cells against PAH-induced DNA damage. The underlying mechanism was mediated by phase I enzyme suppression rather than phase II enzyme induction or oxidative DNA repair. PMID- 19811695 TI - Long-term dietary patterns and carotid artery intima media thickness: the Cardiovascular Risk in Young Finns Study. AB - A whole-diet approach has proven useful for characterising dietary exposure in cardiovascular epidemiology research. In our previous analyses, we found dietary patterns to be significant determinants of CVD risk factor levels among the Cardiovascular Risk in Young Finns cohort. We investigated the associations of major dietary patterns with carotid intima media thickness (IMT), a subclinical predictor of CVD, in healthy adults. The Young Finns Study is an ongoing, prospective cohort study with a 21-year follow-up to date. The subjects were children and adolescents at baseline in 1980 (aged 3-18 years), and all had reached adulthood by the latest follow-up in 2001 (aged 24-39 years). Complete dietary data from the years 1980, 1986 and 2001 and outcome data from the year 2001 were obtained from 785 subjects. The long-term average pattern score for a traditional dietary pattern (characterised by high consumption of rye, potatoes, butter, sausages, milk and coffee) was associated with IMT especially among subjects with a low score for the health-conscious dietary pattern (characterised by high consumption of vegetables, legumes and nuts, rye, tea, cheese and other dairy products). In multivariable regression analyses using long-term pattern scores as predictors, the traditional dietary pattern was independently associated with IMT in men (P < 0.01), but not in women (P = 0.66). Long-term adherence to traditional food choices seems to increase the risk of developing subclinical atherosclerosis among Finnish men. PMID- 19811696 TI - Dietary intake and urinary excretion of lignans in Finnish men. AB - Intake of lignans has been assessed in different study populations, but so far none of the studies has compared the daily intake of lignans and the urinary excretion of plant and enterolignans. We assessed the intake of lariciresinol, pinoresinol, secoisolariciresinol and matairesinol in 100 Finnish men consuming their habitual omnivorous diet, and measured the 24 h urinary excretion of plant and enterolignans to compare the intake and metabolism. Dietary determinants of lignan intake and their urinary excretion were also determined. The mean intake of lignans was 1224 (sd 539) mug/d, of which lariciresinol and pinoresinol covered 78 %. Almost half (47 %) of the intake of lignans was explained by the intake of rye products, berries, coffee, tea and roots. The urinary excretion of plant lignans corresponded to 17 % and enterolignans to 92 % of the intake of lignans. The urinary excretion of plant lignans was explained 14 % by the intake of rye products and intake of coffee, and consequently 3-7 % by the intake of water-insoluble fibre. The urinary excretion of enterolactone was explained 11 % by the intake of vegetables and rye products, 14 % by the intake of water-soluble fibre and only 4 % by the intake of lariciresinol. Although the assessed intake of lignans corresponded well with the urinary excretion of lignans, the enterolactone production in the human body depended more on the dietary sources of lignans than the absolute intake of lignans. PMID- 19811697 TI - Serovars of Leptospira isolated from dogs and rodents. AB - We determined the frequency of isolation of Leptospira from dogs and rodents, the serovars of Leptospira, and the clinical, gross and histological manifestations in dogs with leptospirosis in Trinidad. From dogs, samples of urine, blood and kidney were collected while only kidney and blood samples of trapped rodents were used. Isolates were cultured and serotyped using a panel of 23 international serovars and monoclonal antibodies. The risk factors for leptospirosis were also determined in owned dogs using a standard questionnaire. Of a total of 468 animals investigated for Leptospira, 70 (15.0%) were positive, comprising nine (18.0%) of 50 suspected canine leptospirosis cases, seven (3.4%) of 207 stray dogs and 54 (25.6%) of 211 rodents. The observation that rodents have a statistically (P<0.05, chi2) higher frequency of isolation emphasizes the importance of rodents as reservoirs of leptospirosis in the country. Copenhageni was the predominant serovar found in 100.0% (7/7), 33.3% (2/6) and 68.5% (37/54) of isolates from suspected canine leptospirosis cases, stray dogs and rodents, respectively. Serovars Icterohaemorrhagiae and Canicola, the two serovars present in the commercial vaccines used locally, were detected in one (1.5%) and zero (0.0%) isolates respectively of the 67 tested. Data provided suggest that the apparent vaccine failure may be a consequence of the fact that the predominant serovar (Copenhageni) detected in sick, apparently healthy dogs and in rodents is not contained in the vaccines used locally to protect dogs against canine leptospirosis. PMID- 19811698 TI - Ultrastructural observations reveal the presence of channels between cork cells. AB - The ultrastructure of phellem cells of Quercus suber L. (cork oak) and Calotropis procera (Ait) R. Br. were analyzed using electron transmission microscopy to determine the presence or absence of plasmodesmata (PD). Different types of Q. suber cork samples were studied: one year shoots; virgin cork (first periderm), reproduction cork (traumatic periderm), and wet cork. The channel structures of PD were found in all the samples crossing adjacent cell walls through the suberin layer of the secondary wall. Calotropis phellem also showed PD crossing the cell walls of adjacent cells but in fewer numbers compared to Q. suber. In one year stems of cork oak, it was possible to follow the physiologically active PD with ribosomic accumulation next to the aperture of the channel seen in the phellogen cells to the completely obstructed channels in the dead cells that characterize the phellem tissue. PMID- 19811699 TI - Support for the mutual maintenance of pain and post-traumatic stress disorder symptoms. AB - BACKGROUND: Pain and post-traumatic stress disorder (PTSD) are frequently co morbid in the aftermath of a traumatic event. Although several models attempt to explain the relationship between these two disorders, the mechanisms underlying the relationship remain unclear. The aim of this study was to investigate the relationship between each PTSD symptom cluster and pain over the course of post traumatic adjustment. METHOD: In a longitudinal study, injury patients (n=824) were assessed within 1 week post-injury, and then at 3 and 12 months. Pain was measured using a 100-mm Visual Analogue Scale (VAS). PTSD symptoms were assessed using the Clinician-Administered PTSD Scale (CAPS). Structural equation modelling (SEM) was used to identify causal relationships between pain and PTSD. RESULTS: In a saturated model we found that the relationship between acute pain and 12 month pain was mediated by arousal symptoms at 3 months. We also found that the relationship between baseline arousal and re-experiencing symptoms, and later 12 month arousal and re-experiencing symptoms, was mediated by 3-month pain levels. The final model showed a good fit [chi2=16.97, df=12, p>0.05, Comparative Fit Index (CFI)=0.999, root mean square error of approximation (RMSEA)=0.022]. CONCLUSIONS: These findings provide evidence of mutual maintenance between pain and PTSD. PMID- 19811700 TI - Disorder-specific automatic self-associations in depression and anxiety: results of The Netherlands Study of Depression and Anxiety. AB - BACKGROUND: Cognitive theory points to the importance of negative self-schemas in the onset and maintenance of depression and anxiety disorders. Hereby, it is important to distinguish between automatic and explicit self-schemas, reflecting different cognitive-motivational systems. This study tested whether patients with a current major depression and/or anxiety disorder are characterized by automatic self-anxious and self-depressive associations and whether these associations are disorder specific. METHOD: Patients (n=2329) and non-clinical controls (n=652) were tested as part of The Netherlands Study of Depression and Anxiety, a multi center, longitudinal, cohort study with patients from different health care settings. Patient groups and non-clinical controls (18-65 years of age) were compared with regard to automatic self-anxious and self-depressive associations measured with the Implicit Association Test. RESULTS: Individuals with an anxiety disorder showed enhanced self-anxious associations, whereas individuals with a depression showed enhanced self-depressive associations. Individuals with co morbid disorders scored high on both automatic self-associations. Although remitted individuals showed weaker automatic self-associations than people with a current disorder, their automatic self-anxious/depressed associations were still significantly stronger than those of the control group. Importantly, automatic self-associations showed predictive validity for the severity of anxious and depressive symptoms over and above explicit self-beliefs. CONCLUSIONS: This study represents the first evidence that automatic self-anxious and self-depressive associations are differentially involved in anxiety disorders and depression. This may help to explain the refractoriness of these disorders and points to the potential importance of automatic self-associations in the development of psychopathological symptoms. PMID- 19811701 TI - Genetic contribution to the relationship between personality and depressive symptoms among older women. AB - BACKGROUND: Prior studies suggest that certain types of personality are at higher risk for developing depressive disorders. This study examined the relationship between old age depressive symptoms and two middle-age personality dimensions, neuroticism and extraversion. METHOD: The present study is part of the Finnish Twin Study on Aging, where altogether 409 female twins who had completed the Eysenck Personality Inventory at the age of 38-51 years were studied for depressive symptoms 28 years later using Center for the Epidemiologic Studies Depression Scale. Logistic regression analysis suitable for dependent data and univariate and Cholesky models for decomposing the genetic and environmental factor were used. RESULTS: Middle age extraversion protected from later depressive symptoms while neuroticism increased the risk. Twin modeling indicated that the association between neuroticism and depressive symptoms resulted from shared genetic risk factors common to both traits. However, a substantial proportion of the genetic vulnerability was specific to old age depressive symptoms and was not shared with neuroticism. Middle age extraversion had no genetic relationship with old age depressive symptoms. CONCLUSIONS: The relationship between middle age neuroticism and old age depressive symptoms is strong but only partly the result of genetic factors that predispose to both neuroticism and depressive symptoms. Extraversion, by contrast, has no genetic relationship with depressive symptoms experienced in old age. PMID- 19811702 TI - A common neural system mediating two different forms of social judgement. AB - BACKGROUND: A wide range of neuropsychiatric conditions, including schizophrenia and autistic spectrum disorder (ASD), are associated with impairments in social function. Previous studies have shown that individuals with schizophrenia and ASD have deficits in making a wide range of social judgements from faces, including decisions related to threat (such as judgements of approachability) and decisions not related to physical threat (such as judgements of intelligence). We have investigated healthy control participants to see whether there is a common neural system activated during such social decisions, on the basis that deficits in this system may contribute to the impairments seen in these disorders. METHOD: We investigated the neural basis of social decision making during judgements of approachability and intelligence from faces in 24 healthy participants using functional magnetic resonance imaging (fMRI). We used conjunction analysis to identify common brain regions activated during both tasks. RESULTS: Activation of the amygdala, medial prefrontal cortex, inferior prefrontal cortex and cerebellum was seen during performance of both social tasks, compared to simple gender judgements from the same stimuli. Task-specific activations were present in the dorsolateral prefrontal cortex in the intelligence task and in the inferior and middle temporal cortex in the approachability task. CONCLUSIONS: The present study identified a common network of brain regions activated during the performance of two different forms of social judgement from faces. Dysfunction of this network is likely to contribute to the broad-ranging deficits in social function seen in psychiatric disorders such as schizophrenia and ASD. PMID- 19811703 TI - Re: Neurology out-patients with symptoms unexplained by disease: illness beliefs and financial benefits predict 1-year outcome. PMID- 19811704 TI - Introduction: Cell adhesion and extracellular matrix molecules in synaptic plasticity. PMID- 19811705 TI - The crosstalk of hyaluronan-based extracellular matrix and synapses. AB - Many neurons and their synapses are enwrapped in a brain-specific form of the extracellular matrix (ECM), the so-called perineuronal net (PNN). It forms late in the postnatal development around the time when synaptic contacts are stabilized. It is made of glycoproteins and proteoglycans of glial as well as neuronal origin. The major organizing polysaccharide of brain extracellular space is the polymeric carbohydrate hyaluronic acid (HA). It forms the backbone of a meshwork consisting of CNS proteoglycans such as the lectican family of chondroitin sulphate proteoglycans (CSPG). This family comprises four abundant components of brain ECM: aggrecan and versican as broadly expressed CSPGs and neurocan and brevican as nervous-system-specific family members. In this review, we intend to focus on the specific role of the HA-based ECM in synapse development and function. PMID- 19811707 TI - Research retreat. PMID- 19811708 TI - Frequency and distribution of endodontically treated teeth. AB - OBJECTIVE: To determine the distribution and pattern of carious teeth involvement in permanent teeth requiring endodontic treatment. STUDY DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: Dental Section of The Aga Khan University Hospital, Karachi, from September 2004 to March 2005. METHODOLOGY: Data was collected from the dental records of patients. Tooth was the unit of evaluation. Variables studied were demographics, tooth type, etiology of endodontic treatment and pattern of caries involvement. Data was analyzed using chi-square test and Fischer's exact test. RESULTS: The total number of patients was 190; total number of teeth involved were 235. Females had more endodontically treated teeth than males. There was no significant difference in the distribution of etiology of endodontic treatment in both genders (p=0.564). An increasing trend of endodontic procedure was observed with the increasing age of patients. Caries was the most common etiological factor leading to endodontic treatment (p = 0.011). Class-II cavity i.e. proximal surface of teeth was the most commonly involved surface in endodontically involved teeth (p < 0.001). More molars had been endodontically treated followed by premolars and anterior teeth. CONCLUSION: Caries was the most common etiologic factor; lower molars were the most commonly involved and lower anteriors the least commonly involved teeth in endodontic procedure. Significant association was observed between etiology and tooth type. Significant association was found between cavity classification and etiology of endodontic treatment with class-II caries being most common. PMID- 19811709 TI - Clinico-pathological profile and outcome of inhalational burns. AB - OBJECTIVE: To determine clinico-pathological profile and outcome of inhalational burns in a specialized burns treatment unit. STUDY DESIGN: Case-series. PLACE AND DURATION OF STUDY: The Department of Plastic Surgery and Burns Centre Unit, Combined Military Hospital, Kharian Cantonment in March 2005. METHODOLOGY: Patients of inhalational burns were included and evacuated within 30 hours of accident to the specialized burns centre after immediate resuscitation. Total Body Surface Area (TBSA) involved in burns was calculated. Complete blood count and renal profile along with serum albumin and total proteins was obtained. Portable chest radiographs and bronchoscopic examination was conducted. Escarotomies were carried and wounds were covered with split thickness skin grafts. Ventilatory support was used as needed. Comparison of the clinico pathological profile of surviving and fatal cases was done for significance using t-test. RESULTS: There were 19 patients of inhalational burns, 8 (42%) of whom expired. The mean percentage of TBSA in 11 surviving patients was 50+/-10.87 and 70+/-15.46 in fatal cases. The mean haemoglobin (Hb) on admission was 15.8+/-1.6 g/dL and after fluid resuscitation it became 11.4+/-1.5 g/dL. The mean Total Leucocyte Count (TLC) in surviving patients was 9.6+/-6.1 x 10(9)/L and 1.5+/-2.3 x 10(9)/L in fatal cases (p=0.001). The mean platelet count of surviving patients was 205+/-63 x 10(12)/L while in fatal cases was 58+/-48 x 10(12)/L (p=0.05). The serum urea levels in surviving patients was 4.3+/-2 mmol/L while in fatal cases was 8.6+/-0.9 mmol/L (p=0.05). The serum creatinine levels were 98.2+/-16.5 micromol/L in the survivor group and 249.5+/-76 micromol/L in the mortality group (p=0.05). The serum total protein in surviving patients was 63+/-8 g/dL while in mortality cases it was 57+/-7 g/L. Serum albumin in the survivor group was 36.7+/ 5 g/L and 35+/-4 g/L in fatal cases. Significant in Hb, protein and albumin levels. All the expired patients had acute respiratory distress syndrome while acute renal failure with multi-organ failure co-existed in 6 patients. CONCLUSION: Inhalational burns injury cases multi-system injury with high mortality. Body area involvement, total leucocyte count, platelet count, serum area and serum creatinine are important indicators of survival. PMID- 19811710 TI - Topical diltiazem hydrochloride and glyceryl trinitrate in the treatment of chronic anal fissure. AB - OBJECTIVE: To compare the symptomatic relief, healing and side-effects of topical diltiazem (DTZ) and glyceryl trinitrate in the treatment of chronic anal fissure. STUDY DESIGN: Randomized controlled trial. PLACE AND DURATION OF STUDY: The Surgical Outpatient Department of Civil Hospital, Karachi, from March 2006 to February 2007. METHODOLOGY: Patients with chronic anal fissure were included in the study and randomized to two groups. One group was administered topical 2% diltiazem hydrochloride and other was given 0.2% glyceryl trinitrate (GTN), perianally twice daily for 8 weeks. Patients with anal fissure due to other diseases like inflammatory bowel disease, malignancy, sexually transmitted diseases, previous treatment with local ointment or surgery; patients who required anal surgery for any concurrent disease like hemorrhoids, pregnant women and patients with significant cardiovascular conditions were excluded. There were four follow-up sessions during the course of treatment. Healing and side-effects were recorded. Analysis was done by SPSS version 10 on intention-to-treat basis. Chi-square was used where appropriate. RESULTS: Eighty patients with symptomatic chronic anal fissure were included in the study and equally divided into two groups. After 8 weeks of treatment healing occurred in 31 of 40 patients treated with diltiazem and 33 of 40 patients treated with GTN (p = 0.576). There were less side-effects with DTZ (n=13) than with GTN (n=29, p < 0.001]. In particular, headache occurred more commonly with GTN (n=27) than with DTZ (n=9, p < 0.0001). CONCLUSION: Diltiazem hydrochloride and glyceryl trinitrate were equally effective in healing chronic anal fissure. Diltiazem caused fewer side-effects particularly headache than glyceryl trinitrate ointment. Diltiazem may be the first-line treatment for chemical sphincterotomy for the chronic anal fissure. PMID- 19811711 TI - Body weight and bleeding pattern changes in women using DMPA-SC. AB - OBJECTIVE: To determine the 1-year efficacy of contraception, changes in bleeding pattern and weight with the use of Depot medroxyprogesterone acetate-subcutaneous injected subcutaneously once every 3 months. STUDY DESIGN: Descriptive case series. PLACE AND DURATION OF STUDY: Gynaecology and Obstetrics Unit, Holy Family Hospital, Rawalpindi, from March 2003 to June 2004. METHODOLOGY: Twenty five patients were selected by purposive sampling and followed up in Holy Family Hospital, Gynaecology and Obstetric Unit for a one-year period using DMPA-SC every three months. Hospital Ethical Committee permission was obtained prior to commencement of the study. Informed written consent was taken. Body weight was measured at baseline and every 3 months thereafter. Bleeding analysis in terms of blood flow and severity of bleeding was also done at 3 months interval using a 5 point scale. RESULTS: DMPA-SC showed 100% efficacy in preventing pregnancy in the 25 patients who were followed up. Mean and SD of age was 34.24+/-3.57 years. Mean and SD of weight was 63.44+/-13.81 kg.There was a mean weight gain of 0.1 kg at visit 1-3 (first 3 months) and an average weight gain of 1.036 kg at the end of the year. There was a trend towards amenorrhea with 56% of the patients included in the category of bleeding less than usual at the end of treatment period. CONCLUSION: DMPA-SC can be used in women desiring reversible contraception with unremarkable weight gain and overall bleeding pattern leading towards amenorrhea. PMID- 19811712 TI - Diagnostic accuracy of Tc-99m-MIBI for breast carcinoma in correlation with mammography and sonography. AB - OBJECTIVE: To evaluate the accuracy of 99mTc-MIBI scintimammography (SMM) in differentiating malignant breast cancer from benign breast mass and in detecting axillary lymph node metastasis in comparison with mammography and ultrasonography. STUDY DESIGN: Comparative cross-sectional study. PLACE AND DURATION OF STUDY: At the Karachi Institute of Radiotherapy and Nuclear Medicine (KIRAN), Karachi, from December 2006 to May 2007. METHODOLOGY: A total of 28 patients (both with breast lumps or/and axillary masses) included were in the study. They underwent clinical examination, mammography and ultrasound imaging followed by planar SMM using a single head detector. All subjects received a 740 1110 MBq bolus injection of 99mTc-Sestamibi. 5-10 minutes and 1 hour delayed images were acquired after the injection. SMM scans were considered positive when there was focal area of increased radiotracer uptake. Qualitative (visual) as well as quantitative evaluation of scans was done and compared with ultrasound and mammography, taking histopathology as Gold standard. Sensitivity, specificity, negative and positive predictive values (NPV and PPV respectively) were determined. RESULTS: There were 22 patients presenting with breast lesions (20 palpable, 2 non-palpable) and 6 patients with axillary lump. Scintimammography accurately predicted malignant lesions in the breast (sensitivity 93.3%, specificity. 71.4%, PPV 87.5%, NPV 83.3%, overall accuracy 86.4%) as well as in patients with axillary metastasis (sensitivity 100%, specificity 66%, PPV 75%, NPV 100%, accuracy 83%). A combination of scintimammography with any other imaging modality provides better results than a single test to detect breast cancer. CONCLUSION: SMM has good diagnostic accuracy in the detection of breast cancer as well as in axillary metastasis in association with mammography and ultrasound. PMID- 19811713 TI - An evaluation of renal biopsy in type-II diabetic patients. AB - OBJECTIVE: To determine the renal damage in type-II diabetic patients, who underwent renal biopsy for impaired renal functions and its role in overall patient management. STUDY DESIGN: Descriptive, cross-sectional study. PLACE AND DURATION OF STUDY: The Kidney Postgraduate Centre, Karachi, Pakistan from January 2000 to May 2005. METHODOLOGY: Histopathological evaluation of 73 patients of type-II Diabetes mellitus were included who underwent renal biopsy. Renal biopsy was performed when a renal disease other than diabetic nephropathy was suspected because of the presence of haematuria, nephrotic syndrome, non-nephrotic proteinuria < 3 gms/day in the absence of retinopathy, rapidly progressive glomerulonephritis and renal insufficiency of unknown origin. On the basis of light microscopy and immunofluorescence, three groups of patients were defined. Group I was characterized by diabetic glomerulosclerosis (DGS) only, group II by the prevalence of vascular changes, while group III had sub-groups IIIa (DGS co existing with nondiabetic renal diseases) and IIIb (non-diabetic renal diseases without DGS). RESULTS: Among the 73 patients studied, 20 (27.3%) had diabetic glomerulosclerosis alone (group I), 17 (23.3%) showed occurrence of vascular changes (group II), and 36 (49.3%) had non-diabetic renal diseases (group III). Mean serum creatinine level was significantly greater in group II than in group I and III (p < 0.001). Amount of proteinuria and the presence of haematuria did not show a statistically significant difference in groups I, II and III. The systolic and diastolic blood pressure was higher in groups II as compared to group I and III (p < 0.001). The percentage of sclerotic glomeruli, tubular injury and interstitial inflammation in group II were significantly greater than group I and III (p < 0.001). CONCLUSION: Type-II diabetic patients undergoing renal biopsy for impaired renal functions constituted a heterogeneous group of renal damage. This study emphasized the usefulness of renal biopsy for determining the pattern of renal damage that would aid in the overall management of the patients. PMID- 19811714 TI - Predictive accuracy of intrapartum cardiotocography in terms of fetal acid base status at birth. AB - OBJECTIVE: To find out the predictive value of intrapartum Cardiotocograph (CTG) in terms of fetal acid base status at birth in women undergoing emergency caesarean section for a suboptimal CTG trace. STUDY DESIGN: Observational study. PLACE AND DURATION OF STUDY: At the MCH Centre, Pakistan Institute of Medical Sciences, Islamabad, from June 2004 to July 2005. METHODOLOGY: All women undergoing emergency caesarean section for a suboptimal intrapartum CTG were recruited in the study. Immediately after the delivery of the baby a segment of umbilical cord was doubly clamped at a distance of 10 cm, and 2 ml of arterial cord blood was taken in a heparinized syringe for arterial blood gas indices analysis. All CTG tracings were reviewed using FIGO guidelines and compared for fetal arterial blood gas indices. RESULTS: Of the 57 patients who underwent cesarean section due to suboptimal CTG, 51 (89%) had suspicious trace while 6 (11%) had pathological trace. Positive predictive value of CTG was 18% for fetal hypoxia, 21% for fetal hypercarbia., 26% for fetal acidosis and 37% for base excess. Predictive value of suspicious trace for similar blood indices was 13%, 13%, 17% and 35% respectively. For pathological trace, predictive value was 50%, 83 %, 100% and 66% and respectively. CONCLUSION: Based on the results, it is concluded, that the suspicious CTG trace has low predictive value in terms of fetal acid base status at birth and needs to be complemented with other diagnostic modalities before undertaking any operative intervention. Pathological CTG on the other hand is highly predictive of fetal acidosis at birth warranting immediate intervention. PMID- 19811715 TI - Shoulder impingement syndrome: outcome of arthroscopic subacromial decompression. AB - OBJECTIVE: To describe the outcome of cases with subacromial impingement syndrome managed with arthroscopic subacromial decompression. STUDY DESIGN: Quasi experimental study. PLACE AND DURATION OF STUDY: Section of Orthopaedics, Department of Surgery, The Aga Khan University Hospital, Karachi from April 2005 to March 2006. METHODOLOGY: Thirty patients with impingement syndrome who underwent arthroscopic subacromial decompression after failed conservative treatment were included. The Constant and Murley shoulder scoring system was used for the clinical assessment of pain and function pre-operatively and 1, 6 and 24 weeks postoperatively. RESULTS: The postoperatively significant improvement was observed in the Constant and Murley scores in all the patients (by mean 40 points, p < 0.01), which was progressive over six months. Lower scores were noted in patients over 40 years of age, with advanced stage of impingement and partial tears of the rotator cuff. CONCLUSION: Arthroscopic subacromial decompression was effective in reducing pain and improving function in the studied patients with subacromial impingement syndrome. PMID- 19811716 TI - Venoarterial PCO2 difference: a marker of postoperative cardiac output in children with congenital heart disease. AB - OBJECTIVE: To determine the relationship between venoarterial carbon dioxide gradient (DeltapCO2) and central venous oxygen saturation (ScvO2) in children after cardiac surgery. STUDY DESIGN: A cohort study. PLACE AND DURATION OF STUDY: The Paediatric cardiac intensive care unit of the Aga Khan University Hospital, Karachi, from June 2006 to May 2007. METHODOLOGY: All children admitted in the paediatric cardiac intensive care after complete repair of congenital heart defect using cardiopulmonary bypass were included in the study. Simultaneous arterial and central venous blood gas samples were obtained from a catheter placed in the artery (either radial or femoral) and superior vena cava respectively. Linear regression analysis was performed between ScvO2 and DeltapCO2. RESULTS: Fifty seven children aged from 5 days to 14 years were included and 272-paired simultaneous arterial and central venous samples were analyzed. Mean venous pCO2 was 47.82+/-9.03 mmHg and mean arterial pCO2 was 40.50+/-9.06 mmHg. One hundred seventy four samples had ScvO2 > 70% with mean DeltapCO2 of 5.44+/-2.55 mmHg and 98 samples had ScvO2 < 70% with mean DeltapCO2 of 9.07+/-3.90 mmHg. With ScvO2 < 70%, 77 samples had DeltapCO2 of > 6 mmHg while only 21 samples had DeltapCO2 of < 6 mmHg (p < 0.001). On the contrary with ScvO2 > 70%, 71 samples had DeltapCO2 of > 6 mmHg and 103 samples had DeltapCO2 of < 6 mmHg. Coefficient of correlation (R2) between 0.340 was ScvO2 and DeltapCO2. CONCLUSION: Elevated DeltapCO2 is practical and can be utilized as a useful adjunct to low ScvO2 in the assessment of low cardiac output syndrome in children after cardiac surgery. PMID- 19811717 TI - Cell death induced by Morarah and Khaltita in hepatoma cancer cells (Huh-7). AB - OBJECTIVE: To compare the combined and isolated growth inhibitory effects of Morarah and Khaltita (herbs) on hepatoma cell lines (Huh-7), through induction of apoptosis or necrosis. STUDY DESIGN: Comparative controlled in-vitro study. PLACE AND DURATION OF STUDY: The Molecular Biology Laboratory, The Aga Khan University, Karachi, from June to December 2006. METHODOLOGY: The growth of hepatoma cell lines (Huh-7) was checked by adding Khaltita and Morarah to the cells before culture in a 24 well plate. Six wells were selected and labeled for each of the four variables (controls, Khaltita, Morarah and mixture). After 2 days, cells were studied under an inverted phase contrast microscope and fields were recorded. Approximately four fields per slide of higher intensity were selected randomly to determine the dead cell density, and the procedure was repeated 10 or more times. Frequency and percentages were calculated for dead or alive cells in controls, Morarah, Khaltita and their mixture. Chi-square was used to compare the qualitative variables. P-values < 0.05 were considered significant. RESULTS: Morarah and Khaltita were found to induce statistically significant (p < 0.001) cell death in hepatoma cell lines (Huh-7). At a magnification of 40x, the controls showed 1% dead cells compared to 91% in Morarah, 83% in Khaltita and 73% in combined mixture of Khaltita and Morarah. At magnification of 20x, the controls showed 4% dead cells compared to 44% in Morarah, 47% in Khaltita and 49% in the combined mixture of Khaltita and Morarah. CONCLUSION: Morarah and Khaltita induced cell death in cultured hepatoma cells (Huh-7). PMID- 19811718 TI - Fisher-Mendel controversy in genetics: scientific argument, intellectual integrity, a fair society, Western falls and bioethical evaluation. AB - This review article aims to discuss and analyze the background and findings regarding Fisher-Mendel Controversy in Genetics and to elucidate the scientific argument and intellectual integrity involved, as well as their importance in a fair society, and the lesson of Western falls as learned. At the onset of this review, the kernel of Mendel-Fisher Controversy is dissected and then identified. The fact of an organizational restructuring that had never gone towards a happy synchronization for the ensuing years since 1933 is demonstrated. It was at that time after Fisher succeeded Karl Pearson not only as the Francis Galton Professor of Eugenics but also as the chief of the Galton Laboratory at University College, London. The academic style of eugenics in the late 19th and early 20th centuries in the UK is then introduced. Fisher's ideology at that time, with its effects on the human value system and policy-making at that juncture are portrayed. Bioethical assessment is provided. Lessons in history, the emergence of the Eastern phenomenon and the decline of the Western power are outlined. PMID- 19811719 TI - Ventricular septal defect following disobliteration of right coronary artery. AB - Reperfusion injury is thought to occur during coronary recanalisation but rarely produces clinically significant effects other than arrhythmia. We report an unusual case of Ventricular Septal Defect (VSD) developing after successful disobliteration of the right coronary artery. In this case clinical, electrocardiographic and biochemical evidence of myocardial injury developed 6 hours after successful percutaneous recanalization of the infarct related artery. A rapidly developing VSD soon became apparent necessitating surgical intervention to repair the defect. Unfortunately the patient died soon after surgery. PMID- 19811720 TI - Klippel-Trenaunay syndrome and radial artery coronary graft spasm. AB - A 75-year-old woman with known diagnosis of Klippel-Trenaunay syndrome presented with acute onset of chest pain, dyspnea and elevated cardiac enzymes. She had triple vessel coronary artery disease on subsequent coronary angiography. Given the unavailability of venous conduits secondary to lower extremity varicosities, coronary artery bypass grafting with radial and internal mammary arterial grafts was carried out. The radial artery graft went into spasm two days later and required intracoronary vasodilators to relieve the spasm. The patient remained hypotensive and finally expired. PMID- 19811721 TI - A polypoid gastric heterotopia of jejunum diagnosed by capsule endoscopy. AB - A 36-year-old lady presented with symptoms of intermittent small bowel obstruction caused by a polyp in the jejunum. CT scan and small bowel enema failed to demonstrate this polyp, and required a small bowel capsule endoscopy to finally reach a diagnosis. The objective of this report is to highlight the role of capsule endoscopy as a diagnostic tool in the small bowel pathology and to report a case of a polypoid gastric heterotopia of jejunum. PMID- 19811722 TI - A case of pseudotumorous form of ascaris. AB - A 35-year-old woman presented with lower abdominal pain and amenorrhoea. CT showed a thick walled lobulated mass with Ascaris adjacent to caecum along with the presence of a left ovarian mass. The peroperative findings were a tubular mass with central tunneling containing an Ascaris lumbricoides. Left ovary showed a haemorrhagic cyst. Biopsy of the mass showed acute on chronic granulomatous inflammation and the worm was found to be female. This was a rare case of Ascaris lumbricoides presenting as a pseudotumorous mass. PMID- 19811723 TI - Perforated ileal duplication cyst: a diagnostic dilemma. AB - Alimentary tract duplications are rare congenital anomalies. We report a case of a 14-year-old boy, who presented with acute abdomen, with features favouring acute appendicitis. Laparotomy revealed perforated ileal duplication cyst. Resection of ileum along with cyst and anastomosis was undertaken, and proximal ileostomy performed. Postoperative recovery was uneventful and ileostomy was reversed successfully. Histopathology revealed the presence of gastric mucosa at the site of perforation. PMID- 19811724 TI - Weber's syndrome with vertical gaze palsy. AB - Weber's syndrome with vertical gaze palsy is rarely reported in literature. We present a case of a 47-year-old female who developed sudden onset of left exotropia, right sided hemiplegia and vertical gaze palsy. Magnetic resonance imaging (MRI) showed multiple infarcts involving both thalami and extending caudally into the midbrain. This case presents the diverse clinical picture following midbrain infarcts. PMID- 19811725 TI - Finger prosthesis: the art of reconstruction. AB - Hand deformities affect aesthetics function of hand severely compromised and also cause psychological disturbances. This report describes the fabrication of a silicone finger prosthesis for a patient after an accident at work. The finger prosthesis was retained by a vacuum effect on the stump. The silicone material Silastic-MDX 44210 was used to provide function and aesthetics. The finger prosthesis offered psychological, functional and rehabilitative advantages for the patient. Restoring the natural appearance with the prosthesis eliminated the trauma generated by the dysfunction and represented an efficient psychological therapy. PMID- 19811726 TI - Gastrointestinal abnormalities in vitamin B12 deficient patients with megaloblastic anemia. PMID- 19811727 TI - Partial lipodystrophy. PMID- 19811728 TI - [Expression and localization of adenovirus-mediated transcriptional factor Runx3 in malignant glioblastoma cells]. AB - AIM: To construct the replicative deficient adenovirus Ad-Runx3 expressing Runx3, and to express it in U251 malignant glioblastoma cells. METHODS: The runx3 gene with a flag tag was amplified by PCR using pCMV5-AML2 as a template, and was confirmed by DNA sequencing. The adenovirus shuttle vector pShuttle-CMV-Runx3 was constructed by introducing runx3 DNA fragment into the sites of Kpn I and Xho I of pShuttle-CMV vector. This recombinant plasmid was linearized by PmeI and electronically transfected into BJ5183 cells to get the recombinant adenovirus vector Ad-Runx3. The recombinant adenovirus expressing Runx3 was infected into U251 malignant glioblastoma cells. The expression of exogenous Runx3 was observed by immonoblotting and its localization was detected by immunostaining using anti Flag tag antibody. RESULTS: The recombinant adenovirus expressing Runx3 with a Flag tag was constructed and infected into U251 glioblastoma cells. The exogenous Runx3 protein was detected only in the nuclei. CONCLUSION: The recombinant adenovirus expressing Runx3 with a Flag tag is constructed successfully, and the Runx3 protein expressed in the nuclei of infected cells. The study laid a foundation for further research of the function of Runx3 in gliocarcinogenesis. PMID- 19811729 TI - [Protective effects of ouabain conjugated peptide from Ph.D.-7 Library on vascular endothelial cell]. AB - AIM: To find one kind of peptide that will conjugate with ouabain and inhibit its biological function, and provide a new treatment strategy for primary hypertension. METHODS: In this study, ouabain was used as a target to screen ouabain conjugated peptide (OCP) from Ph.D.-7 phage display peptide library. After 3 rounds of bio-panning, the products were identified by ELISA and DNA electrophoresis analysis and sequencing. Peptide was synthesized and analyzed the activity by radioligand binding assay. The inhibitory ratio of cell proliferation was measured by MTT and the cell morphology changing was measured by Hoechst 33342/PI staining. The expression of Na(+)-K(+)-ATPase alpha1-subunit and beta1 subunit were detected by RT-PCR and immunocytochemistry. The levels of the free intracellular Na(+) in EAhy926 cells were measured by laser confocal microscope. RESULTS: The ouabain conjugated peptide was found out, and it was occupied in 0.64 (9/14). The analysis of protein showed that the obtained peptides had no homology with Na(+)-K(+)-ATPase. The amino acid sequence of synthesized peptide was Arg-Cys-Met-Thr-Ser-Arg-Ser. There was binding activity between OCP and (3)H ouabain. The MTT assay showed that OCP could reverse the inhibition action of ouabain on vascular endothelial EAhy926 cells in a dose and time-dependent manner. The number of apoptotic cells had significantly decreased detected by Hoechst 33342/PI staining. The results of RT-PCR and immunocytochemistry showed that OCP could inhibit the up-regulated expression of Na(+)-K(+)-ATPase alpha1 subunit and down-regulated expression of Na(+)-K(+)-ATPase beta1-subunit induced by ouabain in EAhy926 cells. CONCLUSION: The OCP could reverse the growth inhibition and death induction of ouabain in EAhy926 cells, which would provide the basis for studying the interaction between ouabain and Na(+)-K(+)-ATPase and explore novel anti-ouabain agents. PMID- 19811730 TI - [The photological function of MPA coated CdTe QDs and their biocompatibility]. AB - AIM: To investigate the CdTe quantum dots coated with MPA and explore its biocompatibility with living cells. METHODS: CdTe quantum dots coated with MPA were prepared in aqueous phase and MPA CdTe QDs were Characterized with TEM, fluorospectrophotometer and ultraviolet spectrophotometer. QDs were Modified with with avidin, purified and prepared as fluorescent probe. LSCM was used to observe the expression of MHCII antigen on PMphi cells, which was labeled by QDs. Cell culture and MTT assays were used to determine the biocompatibility of MPA coated CdTe quantum dots with the B-16 cells as target cells. RESULTS: The particle diameter of CdTe quantum dots prepared in aqueous phase was well distributed. They had good photological performance and greater stability after coated with MPA. MHCII antigen on PMphi was labeled with the QDs-Avidin fluorescent probe showed great fluorescence intensity, which was easy to be detected by fluorescence microscope and LSCM. MPA CdTe QDs showed cytotoxicity when its density was very high, but they showed little cytotoxicity during the normal use of influence label density limit. CONCLUSION: MPA CdTe QDs can be used as new fluorescent label as they are of even size, not easy to bleach or quench, have good photological performance and stability and good biocompatibility. PMID- 19811731 TI - [The apoptosis of mouse macrophage J774A.1 induced by oxysterol depend on NF kappaB activation]. AB - AIM: Investigated the relationship between NF-kappaB activation and cell apoptosis in mouse macrophages treated with 7-ketochesterol (7-KC). METHODS: Cell apoptosis was detected by MTT assay, DNA fragmentation assay and flow cytometric analysis. NF-kappaB activation was detected by western blot and immunohistochemistry. Inhibitory assay was used to show the effect of the activation of NF-kappaB on the apoptosis induced by 7-KC. RESULTS: 7-KC inhibited macrophages proliferation, and then induced apoptosis, which is associated with NF-kappaB activation. Moreover, cell apoptosis with NF-kappaB activation was inhibited by pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB. CONCLUSION: 7-KC induced the activation of NF-kappaB and following cell apoptosis. PMID- 19811732 TI - [Cell proliferation inhibited by TIP-6 through autophagy in human hepatoma cell line HepG2 and human normal hepatocyte cell line L02]. AB - AIM: To investigate the effect of 7-(4-methoxyphenyl)-5, 8a-diphenyl-1, 2, 3, 7, 8, 8a-hexahydroimidazo[1, 2-a]pyridine (TIP-6) on cell proliferation in human hepatoma cell line HepG2 and human normal hepatocyte cell line L02. METHODS: Typan blue assay was used to check the effect of TIP-6 on cell proliferation. The changes of cell morphology were observed by the phase contrast microscope. Flow cytometry (FCM) was used to check cell cycle. Autophagy and autophagic cell death were detected after acridine orange (AO) staining under fluorescent microscopy. Apoptosis was analyzed by Annexin V/7-AAD, DAPI staining and DNA ladder. NF kappaB expression was detected with cellular immunochemistry. RESULTS: Cell proliferation inhibiting effect was appeared when treated with TIP-6 from 60 mumol/L to 200 mumol/L, which was correlated with treated concentrations and time. The proliferation rates were just 12.10% and 18.75% (vs control) under 200 mumol/L 72 h in HepG2 and L02 respectively. Vacuolization were found more and more frequently with the increasing of TIP-6 concentrations and treated time prolonged. FCM results indicated that cells were blocked in G2/M phase, and more sensitive were found in HepG2 than L02. AO staining results indicated that the phenomenon of autophagy and autophagic cell death were occurred and appeared more potent with more TIP-6 and longer time treated. No apoptosis markers were found with Annexin V/7-AAD and DAPI staining, and no DNA ladders were found either, these indicated that TIP-6 didn't induce apoptosis in these cells. NF-kappaB was found increased after treated with TIP-6, and the autophagic vacuole became more and more with the increasing of NF-kappaB protein, but the proliferation rates decreased at the same time. CONCLUSION: TIP-6 inhibited cell proliferation and induced autophagy and autophagic cell death in HepG2 and L02 cells. NF-kappaB activation may be involved in these effects. PMID- 19811733 TI - [Immunoregulatory effects of periplocin from Cortex Periplocae in tumor-bearing mice]. AB - AIM: To investigate the immune mechanisms for Periplocin from Cortex Periplocae (CPP) in tumor-bearing mice. METHODS: H(22) tumor-bearing model BALB/c mice were applied to evaluated in vivo immunoregulatory effect of CPP. The influence of different dose CPP (0.25, 0.50 and 1.00 mg/kg) on immune organs in tumor-bearing mice were observed. T cell subsets of mice spleen were detected by flow cytometry. MTT assay was used to determine the influence of CPP on lymphocyte proliferation of mice spleen stimulated by ConA. The levels of TNF-alpha, IL-2 and IL-12 in serum from mice were detected by means of ELISA. RESULTS: Thymus index and spleen index of H(22) tumor-bearing model control mice became less than that of normal mice (P<0.05). Compared to both model and normal control groups, thymus index and spleen index of H(22) tumor-bearing mice treated with CPP increased obviously (P<0.05). CPP had no influence on the number of CD8(+) T cells, but up-regulated markedly the number of CD3(+), CD4(+) T cells and the ratio of CD4(+)/CD8(+) in tumor-bearing mice. In CPP-treated mice, the percentage of CD3(+), CD4(+) T cells were not different from normal mice (P<0.05), the ratio of CD4(+)/CD8(+) was higher than that of normal mice (P<0.05). CPP enhanced obviously lymphocyte proliferation of mice spleen induced by ConA, the SI scores were even higher than that of normal mice. The levels of TNF-alpha, IL-2 and IL 12 in serum from CPP-treated mice, increased significantly compared to model control group (P<0.05) in a dose-dependent manner, were similar to or higher than that of normal mice. CONCLUSION: CPP protected immune organs of tumor-bearing mice, increased obviously the percentage of CD4(+) and CD4(+)/CD8(+) among the T cell line, and enhanced lymphocyte proliferation of mice spleen significantly, stimulated the production of TNF-alpha, IL-2 and IL-12. The results suggested that CPP possessed potent immunoregulatory effect. PMID- 19811734 TI - [Biological effects of lipopolysaccharide, transforming growth factor-beta1 on murine bone marrow-derived dendritic cells]. AB - AIM: To explore method of stimulating murine bone marrow-derived dendritic cells by lipopolysaccharide (LPS), transforming growth factor-beta1 (TGF-beta1) and to study their biological character. METHODS: Murine bone marrow-derived dendritic cells were cultivated with cytokine GM-CSF and IL-4 for 6 days, BMDC was stimulated by control, LPS, TGF-beta1, LPS +TGF-beta1 for 48 hours respectively.Morphological characters of BMDC were observed by a inversed microscope, surface molecules such as CD(11C), CD(80), CD(86) and MHC II were detected by flowcytometry, Interleukin-6 and interleukin-12 p70 in co-culture medium was quantified by ELISA. RESULTS: In LPS group it presented the most typical DC morphology with the highest expression of CD(80), CD(86) and MHC II, the strongest ability in mixed lymphocyte reaction, higher level of IL-6 and IL 12 p70 compared with control, TGF-beta1, LPS+TGF-beta1 (P<0.05). While in TGF beta1 group it presented the less typical DC morphology with the lower expression of CD(80), CD(86), MHC II, weaker ability in mixed lymphocyte reaction, and lower levels of IL-6 and IL-12 p70 compared with control and LPS (P<0.05). CONCLUSION: LPS can stimulate maturation of BMDC in its late differentiation which makes it presents a more significant biological characteristics.TGF-beta1 can inhibit maturation but not differentiation of BMDC thereby can prevent its biological characteristic presentation. PMID- 19811735 TI - [The study of changes on NKT cells of experimental autoimmune encephalomyelitis (EAE) mice]. AB - AIM: To observe the changes of the number of NKT cells in spleens and livers of induced model of experimental autoimmune encephalomyelitis (EAE), and to study the role NKT cells play in the immunoregulation of EAE. METHODS: C57BL/6 mice were immunized with MOG (35-55) peptide and received clinical evaluation daily. The mice were sacrificed at the fastigium and the splenic and hepatic lymphocytes were isolated. The changes of NKT cells in normal and EAE C57BL/6 mice were detected by flow cytometry. RESULTS: The percent of NKT cells in lymphocytes of different organs of EAE model were greater decreased than in that of normal mice. The percent of NKT cells in splenic lymphocytes of normal mice was 2.22+/-0.14, while that in EAE mice was 1.94+/-0.07 (P<0.05). The percent of NKT cells in hepatic lymphocytes of normal mice was 5.52+/-2.17, while that in EAE mice was 2.67+/-1.41 (P<0.05). CONCLUSION: The proliferation of splenic and hepatic NKT cells in C57BL/6 mice are inhibited in EAE model, which may indicate that the immune function conducted by NKT cell is down regulated in EAE mice. PMID- 19811736 TI - [Expression and functional analysis of CRT-E2-EGFP fusion protein in B16 cells]. AB - AIM: To detect the expression of the Calreticulin and HPV E2 Fusion Protein in B16, and study the effects on proliferation and apoptosis of B16 cell lines in vivo. METHODS: To construct eukaryotic fluoresce expression vector pEGFP-CRT-E2, pEGFP-CRT and pEGFP-E2. Then the recombinant plasmids were transfected into B16 cells by Lipofectamine 2000. The expression of proteins was detected by Western blot. The location of different GFP fusion proteins in B16 was tested by inverted fluoresce microscope. Flow cytometry was applied to detect the effects of fusion proteins on the growing of B16 and then the apoptosis effects of B16 induced by different proteins were observed. RESULTS: The correctly constructed recombinant plasmid pEGFP-CRT-E2 and the expression of CRT-E2 gene could be detected in B16 cells. Apoptosis of B16 cells could be detected after the transient transfection. Meanwhile, the apoptosis rate of B16 cells transfected by pEGFP-CRT-E2 was much higher than that of control cells. And cell cycle G1/G0 arrest was also found (P<0.01). CONCLUSION: The eukaryotic expression plasmid pEGFP-CRT-E2 is successfully constructed and it could correctly express the fusion protein in B16 cells. And the B16 cells transfected by plasmid pEGFP-CRT-E2 could induce apoptosis. PMID- 19811737 TI - [Comparison of the effect of arsenic sulfide on apoptosis and hTERT-mRNA expression in two leukemia cell lines]. AB - AIM: To explore the different effect and mechanism of arsenic sulfide on telomerase activity and hTERT-mRNA expression in CML cell lines-K562 and APL cell lines-NB4. METHODS: Telomerase activity was determined by polymerase chain reaction enzyme-linked immunoassay (PCR-ELISA). The expression of hTERT-mRNA was analyzed by semi-quantitative RT-PCR. Flow cytometry was used to analyze the cell cycle and apoptosis. RESULTS: 0.15-0.6 mg/L arsenic sulfide (72 h)can induce apoptosis and inhibit telomerase activity and hTERT-mRNA expression in NB4 cell. The concentration of arsenic sulfide with the same effect on K562 cell was 0.3-3 mg/L. 0.3 mg/L arsenic sulfide (72 h) can cause the proportion of the NB4 cell in G2/M phase increased, but for K562 cell, The concentration of arsenic sulfide was 1.5 mg/L. CONCLUSION: Telomerase system may be one of the pathway for arsenic sulfide inducing apoptosis of NB4 and K562 cell; G2/M phrase arrest may have correlation with decrease of telomerase activity; The sensitivity of NB4 and K562 cell for arsenic sulfide is different, the mechanism of it need to study more. PMID- 19811738 TI - [Effects of recombinant soluble MICA protein on the biologic activities of NK cells]. AB - AIM: To study the effects of recombinant soluble MHC class I chain-related protein A (sMICA) on the cytotoxicity, secretion of IFN-gamma, proliferation and apoptosis of peripheral NK cells. METHODS: After NK cells were co-cultured with recombinant soluble MICA proteins overnight, the cytotoxicity of NK cell on target cells was detected by flow cytometry. The supernant was collected to determine the concentration of IFN-gamma by ELISA. The proliferation of NK cells to sMICA was detected by MTS/PMS. NK cells were labeled with annexin V and PI to analyze their apoptosis. RESULTS: Soluble MICA inhibited the cytotoxicity of NK cells and down-regulated the secretion of IFN-gamma, but it showed no effects on the proliferation and apoptosis of freshly isolated peripheral NK cells. CONCLUSION: The soluble MICA shedding from tumor cells could be a pathway of cancer immune evasion by down-regulating the biologic activities of NK cells. PMID- 19811739 TI - [Immunogenecity of cytomegalovirus-infected fibroblasts]. AB - AIM: To analyze the capability of cytomegalovirus (CMV)-infected human embryonic lung fibroblasts (HELFs) to induce immune response. METHODS: HELFs were infected with cytomegalovirus and stained with antibody against HLA-A2 molecular, the expression of HLA-A2 was detected by FCM. The infected HELFs were incubated with individual pp65 peptide NLVPMVATV. While the uninfected and unloaded infected HELFs served as control respectively. After PBMC was added to the differently treated HELFs and incubated, the immune response was measured with IFN-gamma release as readout. RESULTS: The expression of HLA-A molecular on infected fibroblasts diminished markedly compared with that on the uninfected. The peptides expressed on the infected HELFs together with those pulsed externally induced a stronger response than the infected HELFs alone. CONCLUSION: Although CMV can down-regulate the expression of MHC I on the infected cells, it can not decrease the capacity of cells to present peptides loaded externally, and therefore still induce immune response to some extent. PMID- 19811740 TI - [Preparation and characterization of a mouse anti-human CD40 mutant monoclonal antibody]. AB - AIM: To prepare and characterize a mouse anti-human CD40 mutant monoclonal mAb. METHODS: Female BALB/c mice of 6-8 weeks old were immunized with CD40 mutant transfectant (L929-CD40mu) as immunogen. The spleen B cells of the mice were fused with Sp2/0 myeloma cells.The hybridoma cells were screened with CD40 mutant transfectant (L929-CD40mu) by FCM. Fast-strip analysis was performed to identify Ig subclass of this mAb. The epitope recognized by this mAb was detected by Bio 5C11 competitive assay. Western blot technique was adopted to identify the mAb. The proliferation of tumor cells in vitro was analyzed by MTT assay and apoptosis of tumor cells in vitro was analyzed by PI-annexin V assay. RESULTS: One hybridoma cell line named 10C5 was obtained, which had the property of secreting anti-human CD40 mutant monoclonal antibody continuously and steadily. This mAb specifically recognized human CD40 mutant molecule and induced the apoptosis of tumor cells in vitro. CONCLUSION: One hybridoma cell line which can secret a mouse anti-human CD40 mutant mAb has been prepared successfully. This mAb can inhibit the growth of tumor cells expressing CD40 mutant and induce their apoptosis in vitro. PMID- 19811741 TI - [Preparation and characterization of a novel functional anti-human CD83 monoclonal antibody]. AB - AIM: To prepare and characterize a novel anti-human CD83 monoclonal mAb. METHODS: Female BALB/c mice of 6-8 weeks old were immunized with CD83 transfectant (L929/CD83) as immunogen. The spleen B cells of the mice were fused with Sp2/0 myeloma cells.The hybridoma cells were screened with CD83 transfectant (L929/CD83 and 293/CD83) by FCM. The biological characteristics of antibody were investigated by rapid isotyping analysis, karyotype analysis, competitive inhibition test and Western blot. RESULTS: One hybridoma cell line named 9D8 was obtained, which had the property of secreting anti-human CD83 monoclonal antibody steadily, This mAb specifically recognized CD83 molecule expressed on human mature DC, activated T cells, and tumor cell line Daudi, myeloma cell line 8226.This mAb can recognize a distinct epitope from commercial mAb (HB15e). CONCLUSION: One hybridoma cell line has been developed successfully, which may lay a foundation for further study on the function of this molecule. PMID- 19811742 TI - [Preparation and identification of polyclonal antibody against protein H1b: the variant of major subunit of human ASGPR]. AB - AIM: To prepare and identify mouse polyclonal antibody against protein H1b, which is the variant of major subunit of human ASGPR. METHODS: H1b specific peptide was synthesized and coupled with keyhole limpet hemocyanin (KLH) for immunization. Then H1b-KLH conjugation was injected into mouse subcutaneously to produce polyclonal antibody. ELISA assay was used to detect the titer of the antibody. Antibody was also identified by Western blot and immunohistochemistry assays. RESULTS: Mouse antibody against H1b was prepared after injection of H1b-KLH conjugation. The titer of H1b antibody was about 1:10(5). Western blot confirmed its high specificity. This antibody could also be used for immunohistochemistry analysis. CONCLUSION: The successful preparation of the polyclonal antibody against protein H1b, which can discriminate the two variants of the major subunit of ASGPR with high specificity, will provide an efficient reagent for further study of the physiologic functions of H1b and its role in the pathogenesis of human disease. PMID- 19811743 TI - [Prokaryotic expression and polyclonal antibody preparation of human spermine oxidase]. AB - AIM: To prepare recombinant human spermine oxidase (SMO) and polyclonal antibody against human SMO by gene recombination techniques. METHODS: Human SMO cDNA was amplified from total RNA of A549 cells through reverse transcription PCR. The cDNA was then cloned into pET-15b to construct SMO prokaryotic expression vector. After transforming, the vector was induced to express recombinant SMO by IPTG in E.coli BL21 (DE(3)). Recombinant SMO was purified by Ni-NTA resin under denaturing condition and then was dialyzed to renature. The enzyme activity of recombinant SMO was analyzed by chemical fluorescent method. SMO polyclonal antibody was prepared by using recombinant human SMO protein purified by polyacrylamide gel electrophoresis as antigen to inoculate rabbit intradermally. The titer and specificity of anti-sera were determined by ELISA, Western blot and Immune Cell Chemistry. RESULTS: Purified and dialyzed recombinant human SMO has the specificity of oxidizing the spermine. The polyclonal antibody has high titer and specificity against human SMO. CONCLUSION: This research established a method for prokaryotic expression, purification and polyclonal antibody preparation of human SMO. The method lays a foundation for the future functional research of SMO. PMID- 19811744 TI - [Clone, prokaryotic expression and preparation of polyclonal antibody of hCD59]. AB - AIM: To construct full length hCD59 eukaryotic and extracellular domain of hCD59 (hsCD59)prokaryotic expression vectors and prepare polyclonal antibody of hCD59. METHODS: cDNA fragments encoding hCD59 and hsCD59 were amplified from human PBMCs by RT-PCR and cloned into the eukaryotic vector pVAX-1 and prokaryotic vector pGEX-KG, respectively. The recombinant fusion protein GST-hsCD59 was expressed in E.coil BL21 induced by IPTG. Then the fusion protein was purified and identified. Polyclonal antibody against hCD59 was prepared by immunizing rabbit with pVAX-1 hCD59 and boosting with GST-hsCD59 fusion protein, and the titer was identified. RESULTS: The recombinant eukaryotic vector pVAX-1-hCD59 and prokaryotic vector pGEX-KG-hsCD59 were successfully constructed. The GST-hsCD59 fusion protein was over-expressed in E.coli BL21 and the relative molecular mass (M(r)) of the expression product was identical with predicted size. The titer of the anti-hCD59 serum was 1:3 200. CONCLUSION: We got the recombinant eukaryotic vector pVAX-1 hCD59, prokaryotic vector pGEX-KG-hsCD59 and rabbit anti-hCD59 polyclonal antibody successfully.These work would be helpful for the further study of the biological function of human CD59. PMID- 19811745 TI - [Significance of the alteration of Th17 cells in patients with chronic lymphocytic thyroiditis]. AB - AIM: To investigate the alteration and its significance of T help 17 cells (Th17) in patients with chronic lymphocytic thyroiditis (CLT). METHODS: Patients were divided into 3 groups: CLT patients with euthyroidism (n=15), CLT patients with hypothyroidism (n=30) and healthy control group (n=20). The ratio of Th17 lymphocytes subpopulations in the peripheral blood were evaluated by technique of flow cytometry. Production of thyroid autoantibody (TPO-Ab, TG-Ab) were measured by electro-chemiluminescence immunoassay (ECLIA). RESULTS: Compared with the healthy control group, in CLT group: The frequencies of Th17 in peripheral blood were found to be significantly higher in patients with CLT than healthy control group (P<0.01); Production of TPO-Ab and TG-Ab markedly increased in CLT patients than healthy control group (P<0.01). There was significant correlation between the positive expression of thyroid autoantibody and the changes of Th17 subpopulations (r=0.50, r=0.43 respectively; P<0.01). CONCLUSION: The frequencies of Th17 cell increased in patients with CLT which may suggest a potential role for Th17 in the progression and happen of CLT. PMID- 19811746 TI - [The effect of arsenic sulfide combined with IFN-alpha on K562 cells]. AB - AIM: To study if the effect of arsenic sulfide combined with IFN-alpha can be increased on K562 cells. METHODS: Telomerase activity was determined by PCR ELISA. Flow cytometry was used to analyze the cell apoptosis. The final concentration of IFN-alpha and arsenic sulfide was 10,000 U/mL and 0.6 mg/L. RESULTS: The rates of apoptosis was 37.8% and 37% in K562 cells treated with IFN alpha or arsenic sulfide alone for 8 days; The rates of apoptosis and inhibition of telomerase activity was 59.9% and 81.2% in K562 cells treated with IFN-alpha and arsenic sulfide simultaneously for 8 days, or 60.37% and 78.8% in K562 cells was treated with arsenic sulfide for 5 days after affected by IFN-alpha for 3 days. 71.3% telomerase activity was inhibited in K562 cells by arsenic sulfide alone for 8 days. CONCLUSION: Combination of arsenic sulfide and IFN-alpha can increase the apoptosis and inhibit the telomerase activity of K562 cells obviously comparing with the two drugs used alone. IFN-alpha maybe promote arsenic sulfide inducing apoptosis of K562 cells. PMID- 19811747 TI - [The effects and mechanisms of Forsythia suspense on the expression of Foxp3 on splenocytes and level of Treg in peripheral blood in severely burnt rats]. AB - AIM: To explore the immunity modulation function of aqueous of Forsythia suspense (AFS) and its possible mechanisms. METHODS: Rats of burned model group were burned with vapor under 3 mpa pressure and 108 degrees C temperature for 8 seconds to achieve deep partial-thickness burn, to make a thirty percent total body surface area (TBSA) burn. The experiment were divided into five groups: CONTROL GROUP: without any treatment; 8PBH group: 8 h after burn; the rats of AFS1 group, AFS2 group and AFS3 group of them were given AFS 5 g/kg, 2.5 g/kg, 1.25 g/kg once a day by Po. pathway for seven days before burns, respectively. Rats were sacrificed before and 8h after burn, The percentage of Treg cells in CD4(+) T cells was detected by flow cytometry (FCM); the expression of Foxp3 mRNA on splenocytes were measured by RT-PCR, and the protein of Foxp3 activity was evaluated by immunohistochemistry staining. RESULTS: Compared with CONTROL GROUP, the expression of Foxp3 mRNA and protein on the splenocytes were upregulated markedly (P<0.01), and the percentage of Treg were significantly increased (P<0.01) in the 8PBH group. AFS1, AFS2 and AFS3 significantly attenuated these increases (P<0.01), which was dose-dependent. CONCLUSION: AFS has immunity modulation function and mechanism of it is corrected with Foxp3 mRNA on splenocytes. PMID- 19811748 TI - [Spinal surgery is here to stay]. PMID- 19811749 TI - [Lumbar fusion surgery for degenerative conditions in Denmark 2005-2006]. AB - INTRODUCTION: The indication and use of lumbar spinal fusion surgery for degenerative conditions is being debated. PURPOSE: The objective of this study was to describe recent trends in lumbar fusion surgery for degenerative conditions in Denmark. MATERIAL AND METHODS: A cohort study using a sample from the Danish National Patient Registry was used. RESULTS: In 2005 and 2006, 2,064 and 891 lumbar fusions were performed in 14 public and 6 private hospitals. The annual activity increased from 993 to 1,118 procedures at public hospitals and from 393 to 510 procedures at private hospitals. Lumbar disc arthroplasty was performed in 196 cases, anterior fusion in 162 case, posterior fusion in 2,309 cases, while combined anterior and posterior fusion were performed in 347 cases. The indications for surgery were based on the diagnosis spondylosis in 245 cases, spinal stenosis in 1,174 cases, discuss degeneration in 947 cases, degenerative spondylolisthesis in 600 cases and non-specific lumbar pain in 45 cases. Posterior fusion was performed in 74.3% and 81.9% in public and private hospitals, respectively. Combined anterior and posterior fusion was performed in 12.1% and 10.1% in public and private hospitals, respectively. CONCLUSION: Lumbar fusion rates for degenerative conditions increased from 2005 to 2006 with variation in operative procedures in relation to diagnoses. We propose to monitor data in a database to improve registration of various fusion techniques for various indications and to support further research. PMID- 19811750 TI - [Percutaneous vertebroplasty with only few complications--a retrospective study]. AB - INTRODUCTION: Percutaneous vertebroplasty (PVP) was introduced in 1984 and has proved to be valuable in the treatment of painful vertebral lesions such as haemangiomas, metastasis, and osteoporotic fractures. According to the literature, the operation provides pain relief with only few and harmless complications. The aim of this study was to evaluate our perioperative experiences with special reference to the safety of the procedure. MATERIAL AND METHODS: A total of 156 procedures were performed in 148 patients with osteoporotic fractures. Both acute and chronic fractures were treated. All patients had plain radiographs of the spine. Magnetic resonance imaging (MRI) or bone scans were performed in case of old fractures or more than one fracture in patients with acute back-pain. At follow-up after three and 12 months, plain radiographs were performed in conjunction with physical examinations and interviews. Data were collected retrospectively. RESULTS: We identified 40.4% cemental leaks (confidence interval (CI) 32.6; 48.5). None of these resulted in neurological symptoms. One patient experienced shortness of breath after the procedure, one had atrial fibrillation, and one patient had an iatrogenic pneumothorax. In all, the frequency of symptomatic complications was 1.9% (CI 0.4; 5.5). CONCLUSION: We believe that PVP is a safe procedure with only few symptomatic complications when performed under local anaesthesia and by a specialist in transpedicular approach. PMID- 19811751 TI - [Tumour-cell dissociation as a prognostic marker in colorectal cancer]. AB - The invasive front of the colorectal carcinoma (CRC) in some cases displays budding, characterized by groups of up to five tumour cells. In 2006, budding was introduced in the Danish Colorectal Cancer Group's (DCCG) and the Danish Society of Pathology and Cytology's (DSPAC) CRC register form. Based on a literature survey, molecular mechanisms that may contribute to budding are reviewed, as is the mode by which this process is registered and its putative clinical significance. Despite diverse modes of budding registration, its significance as a prognostic marker has consistently been substantiated. Additionally, data on budding may prove of value in patient management. PMID- 19811752 TI - [Drug-drug interactions in intensive care patients]. AB - INTRODUCTION: The purpose of this study was to investigate the frequency of potential drug-drug interactions (DDIs) within the first 24 hours of admission to an intensive care unit, and to determine which drugs were involved in potential DDIs along with the clinical relevance of the identified DDIs. METHODS: Drug data from all intensive care patients (n = 102) admitted during a three-month period in the spring 2007 were investigated. Potential DDIs were evaluated using the drug interaction system of Micromedex. RESULTS: Four patients were excluded because their treatment only included one drug. A total of 98 patients were treated with an average of ten drugs per patient. Among the enrolled patients, we found 242 potential DDIs, corresponding to an average of 2.5 DDIs per patient. The drugs most frequently involved in potential DDIs included antithrombotic drugs, opioids, loop diuretics, ACE inhibitors, beta blockers, NSAIDs, corticosteroids, quinolon antibiotics, cardiac glycosides, thiazide diuretics, anaesthetics, antidepressants, anticonvulsants and sedatives. DISCUSSION: The number of DDIs per patient found in this study is high compared with the results of a recent Norwegian study. The majority of the identified DDIs were normal combinations of drugs, which are managed through monitoring of the patient and discontinuing of the offending drug if necessary. It is therefore important to revise the patient's medication daily. CONCLUSION: A total of 71% of the 98 enrolled patients were exposed to one or more potential DDIs. We found an average of 2.5 potential DDIs per patient. Antithrombotic drugs, opioids and loop diuretics were most frequently involved in potential DDIs. The clinical relevance varied because the majority of the identified potential DDIs were normal drug combinations. PMID- 19811753 TI - [Egg-donation in Denmark]. AB - Egg-donation is an efficient treatment of female infertility caused by ovarian failure. Although the use of volunteer donors has been allowed in Denmark since 1 January 2007, there is a shortage of oocyte donors, which is in contrast to the increasing demand from couples in need of donor eggs. In this paper we discuss possibilities to enhance the recruitment of volunteer donors. In order to increase the number of egg-donations, the Danish legislation should be changed to allow the use of crossed anonymous donation. PMID- 19811754 TI - [Foetal reduction--a retrospective survey]. AB - INTRODUCTION: Foetal reduction is a procedure by which one or more foetuses in a multi-foetal pregnancy are terminated selectively by an intracardial injection of potassium chloride. There is no Danish study regarding the outcomes after this procedure. The purpose of this study was to calculate results of the foetal reductions performed at Aarhus University Hospital, Skejby. MATERIAL AND METHODS: The study comprised retrospective calculation of 70 foetal reductions performed from 5 June 2000 to 15 June 2007. RESULTS: In all, 44 multiple pregnancies were reduced to two foetuses, which resulted in two total foetal losses (5%), one stillborn and 82 live born, among which two died perinatally. The median gestational age was 255 days (36 weeks and three days), five children were born before week 32 (6%), the mean birth weight among live born was 2,390 +/- 578 grams (mean +/- standard deviation) and 20 were growth restricted (24%). Among 16 pregnancies reduced from two fetuses to one, and ten reduced from three to one, neither miscarriages nor stillbirths were observed. One gave birth before week 32 and the mean birth weight was 3,041 +/- 462 grams. There were no cases of growth restriction. CONCLUSION: The results from Aarhus University Hospital, Skejby, correspond well with the international standard. By reducing to a singleton instead of twins, better results are achieved regarding gestational length, birth weight and growth restriction. PMID- 19811755 TI - [Pharmaceutical treatment of cancer-associated fatigue--a survey of a Cochrane review]. AB - The data and the conclusions of the Cochrane review of drug therapy for the management of cancer related fatigue (CRF) is analysed and summarized in this report. Overall, data is weakened substantially by a high degree of heterogeneity that stems primarily from the fact that fatigue is a subjective and multidimensional phenomenon Apart from erythropoietin and to some degree methylphenidate, there is no evidence that pharmacologic intervention may reduce CRF. CRF has major impact on quality of life for most patients suffering from a cancer disease. It is therefore of obvious importance to increase the focus on CRF - both in the pharmaceutical industry and in the professional-medical environment. PMID- 19811756 TI - [Use of D-dimer to exclude upper extremity deep venous thrombosis]. AB - D-dimer is often used to exclude deep venous thrombosis, primarily in the lower extremities. We describe a 38-year-old man who had deep venous thrombosis in the left vena subclavia in spite of a normal D-dimer. Only one inconclusive survey compares the value of D-dimer and upper extremity deep venous thrombosis. We conclude that where deep venous thrombosis of the upper extremities is suspected, you cannot rely on the D-dimer value, but should examine the patient using other modalities such as colour Doppler ultrasound. PMID- 19811757 TI - [Sudden unexpected death--hypertrophic cardiomyopathy--genetically verified post mortem]. AB - Hypertrophic cardiomyopathy (HCM) may have sudden death as its first presentation. This case presentation describes a 25-year-old man with post-mortem finding of previously unknown left ventricular hypertrophy. Genetic analysis revealed a mutation in the myosin-binding protein C (MYBPC3). Autopsy combined with molecular genetic screening for mutations may give the relatives certainty of cause of death and the opportunity for genetic screening for diagnosis and treatment as well as prevention of sudden cardiac death. PMID- 19811758 TI - [Dermatofibrosarcoma protuberance]. AB - A case of a 33-year-old Caucasian with asymptomatic subdermal dermatofibrosarcomal protuberance of the forehead. Surgical excision with three cm margin and early reconstruction with intraoperative haematurea catheter expansion reducing the defect by 50% and subsequent full thickness skin transplantation of the forehead bone was performed. The case stresses the importance of pathological evaluation of any resected tissue and supports the benefit of intraoperative tissue expansion of the forehead. PMID- 19811759 TI - Miles to go... PMID- 19811760 TI - InfantSEE as a portal to early intervention for autism spectrum disorders. PMID- 19811761 TI - Sex- and gender-based differences in healthy and diseased eyes. AB - PURPOSE: The aim of this study was to identify sex- and gender-based differences in ocular anatomy, physiology, and disease susceptibility or manifestation. METHODS: Review of current indexed literature was conducted. RESULTS: Sex and sex hormones influence the lacrimal system, eyelids and blinking, corneal anatomy and disease, aqueous humor dynamics and glaucoma, crystalline lens and cataract, uveitis and retinal disease, ocular circulation, and optic nerve anatomy and disease. Systemic conditions, particularly autoimmune disease, and conditions that are unique to women, such as pregnancy and menopause, further illustrate the effects of sex hormones on the eye. Gender-based differences in ocular conditions and disease should be considered within the context of the underlying physical and social environment. CONCLUSIONS: Many sex- and gender-based differences exist in healthy and diseased eyes. PMID- 19811762 TI - Idiopathic juxtafoveal retinal telangiectasia. AB - BACKGROUND: Idiopathic juxtafoveal retinal telangiectasia (IJRT) is a rare retinal condition that can be detected even in asymptomatic patients. The most common appearance, type 2A, typically presents as juxtafoveolar telangiectasias with minimal exudation, superficial retinal crystalline deposits, and right-angle venules. Though initially asymptomatic, late complications such as foveolar atrophy and subretinal neovascularization can develop and should be recognized and treated accordingly. CASE REPORT: Two patients with IJRT are presented. The first patient, a 60-year-old Hispanic man, had asymptomatic type 2A IJRT with subretinal neovascularization diagnosed for which he underwent photodynamic therapy on 2 separate occasions. Currently, his visual acuity remains stable at 20/30 in the treated eye. The second patient, a 78-year-old man, had bilateral type 2A IJRT with decreased vision. Fluorescein angiography found no active leakage, and optical coherence tomography found significant foveal atrophy. Therefore, no treatment was advised, and the patient remains with acuities of 20/100- in the right eye and 20/80- in the left. CONCLUSION: Initially, retinal signs can be subtle in type 2A IJRT, with no treatment being indicated. In the late stages of the disease, laser photocoagulation may be necessary to treat subretinal neovascular membranes, which can result as a complication, leading to dramatic vision loss if the fovea is affected. Careful inspection, including the use of fluorescein angiography and ocular coherence tomography, is often helpful in monitoring these patients. PMID- 19811763 TI - Effect of combat eye protection on field of view among active-duty U.S. military personnel. AB - BACKGROUND: This study was conducted to determine if ballistic protective eyewear issued to U.S. military service members causes functional blind spots in the visual field. The study also investigated whether the use of corrective lenses with these devices contributes to defects in the visual field. METHODS: Visual fields were measured using the Esterman Binocular Function test on the Humphrey Field Analyzer. Each subject performed the test with no devices and then repeated the test while wearing several combinations of ballistic eyewear (spectacles or goggles) and prescription correction. Mean differences in Esterman scores between each device and baseline were analyzed. RESULTS: The spectacle devices did not show significant differences in mean Esterman scores from baseline. The goggle systems did show significant reductions in the peripheral field (P < 0.0001). The use of prescription correction with combat eye protection increased the number of missed points in the periphery by an average of 12.2% compared with ballistic devices alone (P < 0.0001). CONCLUSION: The use of ballistic protective goggles and prescription lens inserts with military combat eye protection causes a significant reduction in the peripheral field. Service members should be educated to replace eye movements with head movements when scanning the periphery. PMID- 19811764 TI - Literature and information in vision care and vision science. AB - The explosion of information in vision care and vision science makes keeping up with the literature and information in the field challenging. This report examines the nature of literature and information in vision care and vision science. A variety of topics are discussed, including the general nature of scientific and clinical journals, journals in vision science and vision care, resources available for searches for literature and information, and issues involved in the evaluation of journals and other information sources. Aspects of the application of citation analysis to vision care and vision science are reviewed, and a new citation analysis of a leading textbook in vision care (Borish's Clinical Refraction) is presented. This report is directed toward anyone who wants to be more informed about the literature of vision care and vision science, whether they are students, clinicians, educators, or librarians. PMID- 19811765 TI - Ocular syphilis associated with syphilitic resurgence in a human immunodeficiency virus-infected man. AB - Optometrists provide an important service to the community, often acting as the first health care provider a patient consults. Many patients will present with ocular complaints that have systemic associations. In addition to diabetes, hypertension, and rheumatologic disorders, optometrists can encounter the ocular manifestations of sexually transmitted diseases such as syphilis. Syphilis is a chronic systemic infection caused by the spirochete Treponema pallidum, which is transmitted primarily through sexual contact. A recent rise in the prevalence and clinical manifestation of syphilis increases the likelihood of its presence in the optometric office. PMID- 19811766 TI - A composite fibrin-based scaffold for controlled delivery of bioactive pro angiogenetic growth factors. AB - The aim of this study was to fabricate and characterize in vitro a novel composite scaffold that, combining good mechanical properties with a controlled and sustained release of bioactive pro-angiogenetic growth factors, should be useful for angiogenesis induction in organs/tissues in which is also necessary to give resistance and mechanical strength. Composite scaffolds, constituted by a synthetic biocompatible material, a poly(ether)urethane-polydimethylsiloxane blend, and a biological polymer, the fibrin, were manufactured by spray, phase inversion technique. During the manufacturing process heparin and heparin-binding growth factors, such as VEGF(165) and bFGF, were incorporated into the fibrin layer. Microscopical examinations showed a homogeneous fibrin layer firmly adherent on top of the synthetic material. Tensile tests highlighted the high elasticity of the composite scaffold and its capability to maintain integrity up to high deformation. VEGF(165) and bFGF release were controlled by fibrinogen concentration, whereas it was not affected by heparin concentration, as revealed by ELISA assay. The biological activity of the released growth factors was maintained as demonstrated by HUVEC proliferation. Finally, scaffolds induced a low monocyte mRNA expression of inflammatory markers (IL-8, L-SEL, LFA-1 and iNOS). In conclusion, the new composite scaffolds, once implanted, providing a co localization and temporal distribution of bioactive VEGF and bFGF in addition to good mechanical properties, may be useful to stimulate new vessels formation in ischemic tissues. PMID- 19811767 TI - Overview on current status of biotechnological interventions on yellow stem borer Scirpophaga incertulas (Lepidoptera: Crambidae) resistance in rice. AB - Yellow stem borer (YSB), Scirpophaga incertulas (Lepidoptera: Crambidae), a monophagous pest of paddy is considered as most important pest of rain fed low land and flood prone rice eco-systems. Breeding of yellow stem borer resistance in rice is difficult owing to the complex genetics of the trait, inherent difficulties in screening and poor understanding of the genetics of resistance. On the other hand, a good level of resistance against the widespread yellow stem borer has been rare in the rice germplasm. Resistance to insects has been demonstrated in transgenic plants expressing genes for delta-endotoxins from Bacillus thuringiensis (Bt), protease inhibitors, enzymes and plant lectins. The performance of insect resistant GM rice in trials in China has been quite impressive. The present review is an attempt to assess the current state of development in biotechnological intervention for yellow stem borer resistance in rice. PMID- 19811768 TI - WITHDRAWN: Diversity in mitochondrial metabolic pathways in parasitic protists Plasmodium and Cryptosporidium. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19811769 TI - Evaluation of antidiabetic activity of polysaccharide isolated from Phellinus linteus in non-obese diabetic mouse. AB - Polysaccharide (PLP) isolated from Phellinus linteus inhibits tumor growth and metastasis by enhancing immune functions of macrophages, dendritic cells, NK cells, T cells, and B cells. Here, we report that PLP can inhibit the development of autoimmune diabetes in non-obese diabetic (NOD) mice. Although 80% of the NOD mice had developed diabetes by 24 weeks of age, none of the PLP-treated NOD mice developed diabetes. The mean blood glucose levels were 110mg/dl in PLP-treated mice and 499mg/dl in control NOD mice. Histological examination of the pancreatic islets revealed that most of the islets isolated from PLP-treated mice were less infiltrated with lymphocytes compared with those of control mice. Spleen cells from diabetic NOD mice could adaptively transfer diabetes into NOD/SCID mice, but those from PLP-treated NOD mice showed delayed transfer of diabetes. PLP inhibited the expression of inflammatory cytokines, including IFN-gamma, IL-2, and TNF-alpha by Th1 cells and macrophages, but up-regulated IL-4 expression by Th2 cells in NOD mice. PLP did not prevent streptozotocin-induced diabetic development in ICR mice. Taken together, these results suggest that PLP inhibits the development of autoimmune diabetes by regulating cytokine expression. PMID- 19811770 TI - Sodium selenite inhibits the expression of VEGF, TGFbeta(1) and IL-6 induced by LPS in human PC3 cells via TLR4-NF-(K)B signaling blockage. AB - Lipopolysaccharide (LPS)-induced TLR4-NF-(K)B signaling plays an important role in the development of prostatic tumors from chronic bacterial prostatic infection. Although many studies support the role of selenium in protecting against the development of prostate cancer secondary to chronic prostatitis, the mechanism of action remains unclear. The aim of our study was to investigate whether selenium inhibits the LPS-induced TLR4 signaling pathway in human prostate cancer PC3 cells. Using real-time quantitative PCR and ELISA analysis, we found that pretreatment with selenium (0.5-5uM) inhibited the LPS-induced expression of TGFbeta(1) and VEGF and production of these cytokines and IL-6 by PC3 cells, but did not alter the expression of TLR4 mRNA. Further experiments using Western blot showed that selenium at 3 and 5uM significantly inhibited the translocation of the NF-(K)B p65 subunit to the nucleus in LPS-stimulated PC3 cells. Our results suggest that low doses of selenium may protect the prostate from prostatitis-induced cancer by inhibiting nuclear translocation of the NF (K)B and the subsequent production of the immunosuppressive cytokine TGFbeta(1), proangiogenic factor VEGF and pro-inflammatory factor IL-6. PMID- 19811771 TI - Repair of abdominal wall defects with bovine pericardium. AB - BACKGROUND: Ventral hernia repair with prosthetic mesh has recurrence rates up to 54% and is contraindicated in the setting of infection. The aim of this study was to provide our experience with acellular bovine pericardium (Veritas collagen matrix; Synovis Life Technologies, Inc., St. Paul, MN) in complex abdominal wall reconstruction where prosthetic mesh had failed or was contraindicated. Between 2005 and 2008, a retrospective review of a single general surgeon's practice identified patients reconstructed with acellular bovine pericardium. Thirty primary or recurrent ventral hernias were treated in 26 patients. All patients presented with either contaminated wounds or failure of a prosthetic mesh material. RESULTS: Hernia size ranged from 20 cm(2) to 600 cm(2) (mean 111 cm(2)). Seven patients had previous hernia repair with prosthetic mesh, and 16 patients had ongoing infection or gross contamination at the time of repair. The mean follow-up was 22 months. The hernia recurrence rate in our series was 19% with no fistula development. CONCLUSIONS: Acellular bovine pericardium's high strength, minimal infection rate, and low cost allow its use in the reconstruction of complex abdominal wall defects. PMID- 19811772 TI - Functional connectivity bias of the orbitofrontal cortex in drug-free patients with major depression. AB - BACKGROUND: The orbitofrontal cortex (OFC) plays a crucial role in emotion processing circuits and should therefore also be included in models of the pathophysiology of major depression. The aim of this study was to compare the functional connectivity of the OFC during emotion processing in patients with major depression and healthy control subjects. METHODS: Twenty-five untreated patients with major depression and 15 healthy control subjects were investigated using a functional magnetic resonance imaging face-matching task. RESULTS: Dorsal anterior cingulate cortex, precuneus, and cerebellum activity showed less connectivity with the OFC in patients than in control subjects. In contrast, functional connectivity between the OFC and the right dorsolateral prefrontal cortex (DLPFC), right inferior frontal operculum, and left motor areas was increased in patients compared with healthy control subjects. CONCLUSIONS: The OFC plays a key role in the pathophysiology of major depression. The observed imbalance of OFC connectivity seems to represent a neural mechanism of the processing bias. From a neurobiological point of view, the uncoupling of precuneus and gyrus cinguli activity from the OFC might be associated with problems in the regulation of self-schemas, whereas the increased connectivity of the DLPFC to the OFC might represent a higher neural response to negative stimuli. PMID- 19811773 TI - Early-life blockade of 5-HT(1A) receptors alters adult anxiety behavior and benzodiazepine sensitivity. AB - BACKGROUND: Early-life stress may affect 5-HT(1A) receptor circuitry, which could result in increased anxiety in later life. An increased anxiety phenotype in 5 HT(1A) receptor KO mice (1AKO) mice has been ascribed to 5-HT(1A) receptor absence during the early postnatal period. Thus, subtle and transient serotonergic changes during the early postnatal period may lead to an increased risk for developing stress-related disorders during adulthood. METHODS: Wildtype and 1AKO mice on a Swiss-Webster (SW) background were treated during the early postnatal period with vehicle or the 5-HT(1A) receptor antagonist WAY-100,635. RESULTS: Pharmacologic 5-HT(1A) receptor blockade during the early postnatal period induced long-lasting effects on anxiety and benzodiazepine sensitivity in adolescent and adult mice on a Swiss-Webster background and resembles the SW 1AKO phenotype. Furthermore, WAY-100,635-treated mice had increased cortical gamma aminobutyric acid-A receptor (GABA(A)R) alpha(1) and alpha(3) subunit levels and increased hippocampal GABA(A)R alpha(2) subunit levels. CONCLUSIONS: Absence of 5 HT(1A)R signaling during early stages of brain maturation predisposes an organism to affective dysfunction later in life. Because early-life treatment with WAY 100,635 in Swiss-Webster mice reduced diazepam sensitivity and increased GABA(A)R alpha subunit levels in the prefrontal cortex and hippocampus, our data suggest a putative link between early-life disruption of the serotonergic system and the emergence of increased anxiety and decreased benzodiazepine responsivity at adult age. Moreover, early-life 5-HT(1A) receptor functionality appears to be essential for the development of normal GABA(A)R functionality. This study may have clinical implications for psychoactive drug use during pregnancy and for the pharmacogenetic background of benzodiazepine sensitivity. PMID- 19811774 TI - Mapping attention-deficit/hyperactivity disorder from childhood to adolescence- no neurophysiologic evidence for a developmental lag of attention but some for inhibition. AB - BACKGROUND: The role of a developmental lag for deficits of higher brain functions in attention-deficit/hyperactivity disorder (ADHD) has not yet been tested in longitudinal studies. We examined the development of neurophysiological markers of attention (Cue P300; contingent negative variation [CNV]) and inhibition (NoGo P300) in ADHD and control groups from childhood to adolescence for support of the developmental lag hypothesis of ADHD. METHODS: ADHD (n = 28/3 girls) and control (n = 22/5 girls) subjects were assessed at baseline (Time 1; ADHD age 10.8 +/- 1.8 years, controls 10.4 +/- 1.1 years) and at two follow-up examinations (Time 2 after 1.2 years, Time 3 after 2.5 years). Event-related potential maps were recorded during a cued Continuous Performance Test (CPT) at all assessments and analyzed using scalp and source (sLORETA) measures. RESULTS: CPT performance showed common effects of ADHD and younger age, consistent with (but not specific to) developmental lag. The NoGo P300 developed earlier and became stronger in control subjects than in the ADHD group, again consistent with an initial developmental lag. In contrast, the attenuation of the Cue P300 and the CNV with ADHD at all assessments was opposite to the enhancement with younger age and thus inconsistent with developmental lag. The sLORETA source localization also differed between ADHD and developmental effects. CONCLUSIONS: These results provide strong evidence for multiple and persistent neural processing deficits in ADHD. They do not support the developmental lag hypothesis for attentional dysfunction in ADHD despite partial evidence that developmental lag contributes to inhibitory brain dysfunction during early adolescence. PMID- 19811776 TI - Efficacy of d-cycloserine for enhancing response to cognitive-behavior therapy for panic disorder. AB - BACKGROUND: Traditional combination strategies of cognitive-behavior therapy plus pharmacotherapy have met with disappointing results for anxiety disorders. Enhancement of cognitive-behavior therapy with d-cycloserine (DCS) pharmacotherapy represents a novel strategy for improving therapeutic learning from cognitive-behavior therapy that remains untested in panic disorder. METHOD: This is a randomized, double-blind, placebo-controlled augmentation trial examining the addition of isolated doses of 50 mg d-cycloserine or pill placebo to brief exposure-based cognitive-behavior therapy. Randomized participants were 31 outpatients meeting DSM-IV criteria for panic disorder with or without agoraphobia, who were offered five sessions of manualized cognitive-behavior therapy emphasizing exposure to feared internal sensations (interoceptive exposure) but also including informational, cognitive, and situational exposure interventions. Doses of study drug were administered 1 hour before cognitive behavior therapy sessions 3 to 5. The primary outcome measures were the Panic Disorder Severity Scale (PDSS) and Clinicians' Global Impressions of Severity. RESULTS: Results indicated large effect sizes for the additive benefit of d cycloserine augmentation of cognitive-behavior therapy for panic disorder. At posttreatment and 1 month follow-up, participants who received d-cycloserine versus placebo had better outcomes on the PDSS and global severity of disorder and were significantly more likely to have achieved clinically significant change status (77% vs. 33%). There were no significant adverse effects associated with DCS administration. CONCLUSIONS: This pilot study extends support for the role of d-cycloserine in enhancing therapeutic learning from exposure-based cognitive behavior therapy and is the first to do so in a protocol emphasizing exposure to feared internal sensations of anxiety in panic disorder. PMID- 19811775 TI - Effects of estradiol on learned helplessness and associated remodeling of hippocampal spine synapses in female rats. AB - BACKGROUND: Despite the fact that women are twice as likely to develop depression as men, our understanding of depression neurobiology in female subjects is limited. We have recently reported in male rats that development of helpless behavior is associated with a severe loss of hippocampal spine synapses, which is reversed by treatment with the antidepressant desipramine. Considering that estradiol has a hippocampal synaptogenic effect similar to those of antidepressants, the presence of estradiol during the female reproductive life might influence behavioral and synaptic responses to stress and depression. METHODS: With electron microscopic stereology, we analyzed hippocampal spine synapses in association with helpless behavior in ovariectomized female rats (n = 70), under different conditions of estradiol exposure. RESULTS: Stress induced an acute and persistent loss of hippocampal spine synapses, whereas subchronic treatment with desipramine reversed the stress-induced synaptic loss. Estradiol supplementation given either before stress or before escape testing of nonstressed animals increased the number of hippocampal spine synapses. Correlation analysis demonstrated a statistically significant negative correlation between the severity of helpless behavior and hippocampal spine synapse numbers. CONCLUSIONS: These findings suggest that hippocampal spine synapse remodeling might be a critical factor underlying learned helplessness and, possibly, the neurobiology of depression. PMID- 19811777 TI - Dopamine type 2/3 receptor availability in the striatum and social status in human volunteers. AB - BACKGROUND: Previous positron emission tomography (PET) imaging studies in nonhuman primates have shown that striatal dopamine type 2/3 (D(2/3)) receptors correlate with social hierarchy in monkeys and that dominant animals exhibit higher levels of D(2/3) receptor binding. The goal of the present study was to examine this phenomena in human subjects using PET and the radiotracer [(11)C]raclopride. METHODS: Fourteen healthy volunteers were scanned with [(11)C]raclopride to measure D(2/3) receptor binding potential (BP). Social status was assessed using the Barratt Simplified Measure of Social Status. In addition, participants were asked to assess their level of social support using the Multidimensional Scale of Perceived Social Support (MSPSS). RESULTS: A correlation was seen between social status and dopamine D(2/3) receptors, where volunteers with the higher status had higher values for [(11)C]raclopride BP. A similar correlation was seen with the perceived social support, where higher [(11)C]raclopride BP correlated with higher scores on the MSPSS. CONCLUSIONS: The results of this study support the hypothesis that social status and social support is correlated with D(2/3) receptor binding. PMID- 19811778 TI - T lineage differentiation from induced pluripotent stem cells. AB - Induced pluripotent stem (iPS) cells have potential to differentiate into T lymphocytes, however, the actual ability of iPS cells to develop into T lineages is not clear. In this study, we co-cultured iPS cells on OP9 cells expressing the Notch ligand Delta-like 1 (DL1), the iPS cells differentiated into T lymphocytes. In addition, in vitro stimulation of iPS cell-derived T lymphocytes resulted in secretion of IL-2 and IFN-gamma. Moreover, adoptive transfer of iPS cell-derived T lymphocytes into Rag-deficient mice reconstituted their T cell pools. These results indicate that iPS cells are able to follow the normal program of T cell differentiation. PMID- 19811779 TI - The contributions of cerebro-cerebellar circuitry to executive verbal working memory. AB - Contributions of cerebro-cerebellar function to executive verbal working memory were examined using event-related functional magnetic resonance imaging (fMRI) while 16 subjects completed two versions of the Sternberg task. In both versions subjects were presented with two or six target letters during the encoding phase, which were held in memory during the maintenance phase. A single probe letter was presented during the retrieval phase. In the "match condition", subjects decided whether the probe matched the target letters. In the "executive condition", subjects created a new probe by counting two alphabetical letters forward (e.g., f-->h) and decided whether the new probe matched the target letters. Neural activity during the match and executive conditions was compared during each phase of the task. There were four main findings. First, cerebro-cerebellar activity increased as a function of executive load. Second, the dorsal cerebellar dentate co-activated with the supplementary motor area (SMA) during encoding. This likely represented the formation of an articulatory (motor) trajectory. Third, the ventral cerebellar dentate co-activated with anterior prefrontal regions Brodmann Area (BA) 9/46 and the pre-SMA during retrieval. This likely represented the manipulation of information and formation of a response. A functional dissociation between the dorsal "motor" dentate and "cognitive" ventral dentate agrees with neuroanatomical tract tracing studies that have demonstrated separate neural pathways involving each region of the dentate: the dorsal dentate projects to frontal motor areas (including the SMA), and the ventral dentate projects to frontal cognitive areas (including BA 9/46 and the pre-SMA). Finally, activity during the maintenance phase in BA 9, anterior insula, pre-SMA and ventral dentate predicted subsequent accuracy of response to the probe during the retrieval phase. This finding underscored the significant contribution of the pre SMA/ventral dentate pathway--observed several seconds prior to any motor response to the probe--to executive verbal working memory. PMID- 19811780 TI - Mortality among frequent consumers of Great Lakes sport fish. AB - Commercial and sport-caught fish provide a healthy source of dietary protein, omega-3 fatty acids, and other micronutrients. Regular fish consumption has been associated with decreased risk of heart disease and health professionals encourage adults to include fish in their weekly diets. However, fish harvested from contaminated waters can contain higher levels of persistent, bioaccumulative chemicals such as methylmercury, PCBs, dieldrin, and DDT. To assess the beneficial effects of fish intake and the adverse effects of contaminant exposure, underlying and contributing causes of death were obtained from the National Death Index for 342 deceased members of a cohort of 2538 Great Lakes charterboat captains, 180 Wisconsin anglers, and 1141 referents who were established in 1993-1995. Multivariate analysis of death rates confirmed a dose related protective effect of fish intake against all-cause and cardiovascular disease mortality, including coronary heart disease, among the referent group. This effect was not observed among consumers of Great Lakes sport fish, however. Cancer mortality was not associated with fish intake in either exposure group. While the number of deaths among this cohort is currently too small to support rigorous statistical analysis, these preliminary findings are consistent with other studies that have shown a protective effect of commercial fish on human health and longevity and raise concerns regarding the effect of persistent environmental contaminants that continue to be detected in fish from the Great Lakes Basin. It is hoped that continued monitoring of this cohort will improve our understanding of the complex interactions that exist between nutrients and contaminants found in fish harvested from the Great Lakes. PMID- 19811782 TI - Design and implementation of an automated secondary cooling system for the continuous casting of billets. AB - This paper describes a heat transfer model based automatic secondary cooling system for a billet caster. The model aims to minimize the variation in surface temperature and excessive reheating of the billet strands. It is also used to avoid the low ductility trough of the solidifying steel, which aggravates the tendency of steel to crack. The system has been designed and implemented in an integrated steel plant. A Programmable Logic Controller (PLC) based automation system has been developed to control the water flow in the secondary cooling zones of the strand. The results obtained through field trials have shown complete elimination of internal and off-corner cracks for the fifty billet samples that were monitored. PMID- 19811781 TI - Contaminant profiles in Southeast Asian immigrants consuming fish from polluted waters in northeastern Wisconsin. AB - Recent immigrants to the USA from Southeast Asia may be at higher risk of exposure to fish-borne contaminants including polychlorinated biphenyls (PCBs), p, p'-dichlorodiphenyldichloroethene (DDE) and methylmercury (MeHg) because of their propensity to engage in subsistence fishing. Exposure to contaminants was assessed in men and women of Hmong descent living in Green Bay, Wisconsin, where the Fox River and lower Green Bay are contaminated with PCBs, and to a lesser extent with mercury. Serum samples from 142 people were analyzed for PCBs and p,p'-DDE by capillary column gas chromatography with electron capture detection (ECD). Whole blood was analyzed for total mercury by cold vapor atomic absorption spectrometry and atomic fluorescence spectroscopy. Lipid-adjusted total PCB concentrations ranged from 8.7 to 3,091ng/g (full range of the data), with a geometric mean of 183.6ng/g (estimated after eliminating one outlier). DDE ranged from 0.3 to 7,083 (full range of the data) with a geometric mean of 449.8ng/g (estimated after eliminating two outliers). Men had higher PCB and DDE concentrations than women. Serum PCB concentrations were significantly correlated with fish consumption (r=0.43, p<0.0001), whereas DDE concentrations were not (r=0.09,p=0.29). Instead, serum DDE was strongly associated with the number of years spent in a Thai refugee camp before immigrating to the USA (r=0.60;p<0.0001). PCB congeners 138, 153, 118 and 180 accounted for a smaller percentage of the total PCBs than has been reported in other fish-eating populations, and several lightly chlorinated congeners were present in relatively large amounts. Mercury exposure was low in this population. In conclusion, Hmong immigrants in northeastern Wisconsin are at risk of elevated PCB exposure from consumption of locally caught fish. The pattern of exposure is somewhat different than patterns in other fish-eating populations, possibly due to use of Aroclor 1242 by the paper industry in this region. PMID- 19811783 TI - A multi-scale approach to understand the mechanobiology of intermediate filaments. AB - The animal cell cytoskeleton consists of three interconnected filament systems: actin microfilaments, microtubules and the lesser known intermediate filaments (IFs). All mature IF proteins share a common tripartite domain structure and the ability to assemble into 8-12nm wide filaments. At the time of their discovery in the 1980s, IFs were only considered as passive elements of the cytoskeleton mainly involved in maintaining the mechanical integrity of tissues. Since then, our knowledge of IFs structure, assembly plan and functions has improved dramatically. Especially, single IFs show a unique combination of extensibility, flexibility and toughness that is a direct consequence of their unique assembly plan. In this review we will first discuss the mechanical design of IFs by combining the experimental data with recent multi-scale modeling results. Then we will discuss how mechanical forces may interact with IFs in vivo both directly and through the activation of other proteins such as kinases. PMID- 19811784 TI - Neural mechanobiology and neuronal vulnerability to traumatic loading. AB - In order to understand the physical tolerance of neurons to traumatic insults, engineers and neuroscientists have attempted to reproduce the biomechanical environment during a traumatic event using in vitro injury systems with isolated components of the nervous system. This approach allows one to begin to unravel the underlying molecular and biochemical mechanisms that lead to cell dysfunction and death as a function of mechanical inputs. Excess mechanical force and deformation causes structural and functional breakdown, including several key deleterious cellular processes, such as membrane damage, an upset of calcium homeostasis, glutamate release, cell death, and caspase-mediated proteolysis. Understanding of the mechanotransduction events, however, that lead to cellular failure and dysfunction, are not well understood. Mechanically characterized cellular models of traumatic loading are critical to the improved understanding of mechanotransduction in the context of neural injury, the improvement of protective systems, and to provide a controlled setting for testing therapeutic interventions. In this review of the cellular mechanics of traumatic neural loading, we focus on the backdrop and motivation for studying mechanical thresholds in neurons and glial cells and discuss some of the acute responses that may help elucidate improved tolerance criteria and illuminate future research directions. PMID- 19811785 TI - Mechano-potential etiologies of aortic valve disease. AB - Aortic valve leaflets experience varying applied loads during the cardiac cycle. These varying loads act on both cell types of the leaflets, endothelial and interstitial cells, and cause molecular signaling events that are required for repairing the leaflet tissue, which is continually damaged from the applied loads. However, with increasing age, this reparative mechanism appears to go awry as valve interstitial cells continue to remain in their 'remodeling' phenotype and subsequently cause the tissue to become stiff, which results in heart valve disease. The etiology of this disease remains elusive; however, multiple clues are beginning to coalesce and mechanical cues are turning out to be large predicators of cellular function in the aortic valve leaflets, when compared to the cells from the pulmonary valve leaflets, which are under a significantly less demanding mechanical loading regime. Finally, this paper discusses the mechanical environment of the constitutive cell populations, mechanobiological processes that are currently unclear, and a mechano-potential etiology of aortic disease will be presented. PMID- 19811787 TI - Porous graphitic carbon: a versatile stationary phase for liquid chromatography. AB - Chromatographic stationary phases based on porous graphitic carbon were invented 30 years ago, while columns have been commercially available for 20 years. This special occasion deserved a complete review on this material. In this paper, we describe current knowledge on graphitic carbon stationary phases, based on over 400 fundamental studies and applications. PMID- 19811788 TI - Recent advances in microextraction by packed sorbent for bioanalysis. AB - Microextraction by packed sorbent (MEPS) is a new format for solid-phase extraction (SPE) that has been miniaturized to work with sample volumes as small as 10muL. The commercially available presentation of MEPS uses the same sorbents as conventional SPE columns and so is suitable for use with most existing methods by scaling the reagent and sample volumes. Unlike conventional SPE columns, the MEPS sorbent bed is integrated into a liquid handling syringe that allows for low void volume sample manipulations either manually or in combination with laboratory robotics. The key aspect of MEPS is that the solvent volume used for the elution of the analytes is of a suitable order of magnitude to be injected directly into GC or LC systems. This new technique is very promising because it is fast, simple and it requires very small volume of samples to produce comparable results to conventional SPE technique. Furthermore, this technique can be easily interfaced to LC/MS and GC/MS to provide a completely automated MEPS/LC/MS or MEPS/GC/MS system. This extraction technique (MEPS) could be of interest in clinical, forensic toxicology and environmental analysis areas. This review provides a short overview of recent applications of MEPS in clinical and pre-clinical studies for quantification of drugs and metabolites in blood, plasma and urine. The extraction of anti-cancer drugs, beta-blockers drugs, local anaesthetics, neurotransmitters and antibiotics from biological samples using MEPS technique will be illustrated. PMID- 19811786 TI - The biomechanical integrin. AB - The integrin lies at the center of our efforts to understand mechanotransduction in the human body. Over the past two decades, a wealth of information has yielded important insights into integrin structure and functioning in biochemical pathways; however, relatively little emphasis has been placed on mechanics. In this article, we review the current knowledge base of integrin mechanobiology by examining the role of integrins in stabilizing tissue structure, the mechanisms of integrin force transfer, the process of cell migration, and the pathology of cancer. In order to successfully address the gaps in cancer and other disease research going forward, future efforts of integrin mechanobiology must focus on examining cells in 3D environments and integrating our current understanding into computational models that predict the behavior of integrins in non-equilibrium interactions. PMID- 19811789 TI - The fabrication of photosensitive self-assembly Au nanoparticles embedded in silica nanofibers by electrospinning. AB - In this study, we demonstrated a simple, efficient, and low-cost method to fabricate large-area self-assembly Au nanoparticles (AuNPs) encapsulated within silica nanofibers (Au/SiO(2)). The method is based on electrospinning and thermal decomposition of hybrid nanofibers prepared from the solution of tetraethylorthosilicate (TEOS), polyvinylpyrrolidone (PVP) and AuNPs. This study employed the electrospinning technique for the first time as a successful method for preparing a self-organized AuNP peapod chain in a silica nanofiber matrix, under mild conditions. It has the advantage of easily controlling the diameters of the silica nanofibers as well as the concentration of the AuNPs in the spinning solution. The Au/SiO(2) hybrid nanofibers fabricated by this method exhibited an obvious photoelectric response under the illumination wavelength around the Au/SiO(2) nanofibers surface plasmon resonance (SPR) absorption band, whereas no photoelectric response was observed for the pure silica fibers. The excellent characteristics of photoelectric response suggest that the electrospinning technique has a great potential for large-scale fabrication of functional nanofiber devices. The ability of coupling light responses into the nanosystems dependent on metallic nanoparticle SPR opens up new prospects for the construction of nanoscale waveguiding devices, sensors and optoelectronics. PMID- 19811790 TI - Hydrogen sulfide protects against ischemia-reperfusion injury in an in vitro model of cutaneous tissue transplantation. AB - BACKGROUND: Ischemia-reperfusion injury (IRI) is a source of morbidity and mortality in many clinical scenarios, and has as one of its many consequences the induction of cellular apoptosis. Hydrogen sulfide (H2S) may decrease cellular metabolism in a reversible, nontoxic manner. An in vitro model of cutaneous tissue transplantation was developed to assess whether H2S could ameliorate cellular injury caused by IRI. METHODS: Human umbilical vein endothelial cells (HUVECs) were treated with media containing NaHS (0, 10 microM, 100 microM, or 1 mM) and exposed to normoxia (21% oxygen), hypoxia (1%), or anoxia (0%). Cells were then returned to normoxia, and apoptosis was quantified using a terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Fibroblasts (3T3s) were treated with H2S and exposed to anoxia in a similar fashion. RESULTS: Treatment with H2S resulted in a significant decrease in apoptosis in HUVECs and 3T3s subjected to IRI. Toxicity of H2S was not observed, although the protective effect was less evident at higher doses. CONCLUSION: This is the first study to examine H2S and the cellular components of cutaneous flaps in the setting of IRI. Our results demonstrate that H2S significantly decreases apoptosis in vitro in the setting of IRI. These data suggest H2S may mitigate IRI in vivo, and, therefore, has potential as a therapy for improving tissue survivability in clinical scenarios. PMID- 19811791 TI - [Common variable immunodeficiency. Prognostic factors for lung damage]. AB - BACKGROUND AND OBJECTIVE: Lung damage is considered to be key to Common Variable Immunodeficiency (CVID) prognosis. We describe lung damage in pediatric CVID patients and assess its relationship with memory B cells (MB) phenotype, immunoglobulin G (IgG) levels at the time of diagnosis, and diagnostic delay. We also assessed the prevalence of allergy and autoimmune phenomena. PATIENTS AND METHOD: Cross-sectional study of 17 CVID patients treated at Vall d'Hebron University Hospital. Prevalence ratio and prevalence odds ratio were used to assess the effect of MB cells phenotype, IgG levels and diagnostic delay on lung damage. RESULTS: Five of seventeen patients presented bronchiectasis. Diagnostic delay >5 years was significantly associated with more severe lung damage. MBO phenotype and low IgG levels at the time of diagnosis showed a trend to more severe lung damage without reaching statistical signification. A higher prevalence of allergic or autoimmune phenomena was not observed. CONCLUSIONS: Latter diagnosis is associated with greater lung damage in CVID patients. The study of MB cells should be included in the study of these patients. PMID- 19811792 TI - [Cured ham and incidence of cardiovascular events, arterial hypertension or weight gain]. AB - BACKGROUND AND OBJECTIVE: Cured ham is a characteristic food of Spanish Mediterranean diet. However, no prospective epidemiologic study assessing its effects on human health is available. Our aim was to assess the association between the consumption of cured ham and cardiovascular disease, hypertension or weight gain. SUBJECTS AND METHOD: In a prospective and dynamic epidemiologic cohort composed exclusively of university graduates (the SUN Project, n=13,293), we analyzed the incidence of cardiovascular disease, hypertension and average yearly weight gain after a maximum follow-up of 6 years. Cox (proportional hazards) regression models were fitted to estimate hazard ratios (relative risks [RR]) after adjusting for potential confounding. RESULTS: No association was found between higher levels of consumption of cured ham (>4 servings/week) and the incidence of cardiovascular disease (RR=1.02; [95%CI]: 0.44-2.39), in analyses adjusted for age, sex, total energy intake and mediterranean dietary pattern, compared to the consumption of less than one serving a week. When we repeated this comparison for the incidence of hypertension, and adjusting for age, sex, total energy intake and mediterranean dietary pattern the RR was 0.74 (95% CI: 0.55-1.01). In the comparison of average yearly weight gain between these extreme categories of cured ham consumption (<1 versus > or =4 servings/week) a non-significant difference of 0.039 kg (95%IC: -0.036 to 0.113) was found after adjusting for sex, age, total energy intake, mediterranean dietary pattern, smoking, physical activity, and baseline body mass index. CONCLUSIONS: The results of this cohort study do not support any association between the consumption of cured ham and a higher risk of cardiovascular disease, hypertension or weight gain. PMID- 19811793 TI - [Contribution of neuroradiology to the evaluation of peripheral nerves]. AB - The evaluation of peripheral nerve impairment can use echography, MRI and CT. The alteration of the nerves in tunnel syndromes, especially if symptoms are not sufficiently contributive, is clearly visualized with MRI. Echography is very useful in the diagnosis of nerve tumors and neuromas. Plexus brachial palsies need to be evaluated using MRI. Progress in neuroimaging has improved clinical practice so that the most relevant treatment can be chosen for some pathologies such as infiltration performed under CT scanner guidance. The authors report which exam to use and the results to be expected for each pathology. PMID- 19811794 TI - [Occlusal plane orientation and postoperative anterior open bite relapse]. AB - INTRODUCTION: Most published data on relapse in open bite maxillo-mandibular deformities give raw results but do not suggest any specific therapy. Indeed, their authors compare the various osteotomy techniques but without identifying risk factors for relapse (dysfunctional or architectural). We studied the predictive value of occlusal plane tilting, in the long-term relapse of open bite maxillo-mandibular deformity. MATERIAL AND METHOD: Fifty patients were included between 1996 and 2007. For each patient, Delaire cephalometric analysis was performed on preoperative, immediate and late postoperative teleradiographs. Immediate real postoperative occlusal plane tilting was analyzed and compared with "ideal" theoretical occlusal plane tilting (calculated with Delaires' analysis). The patients were classified in two groups: one with slight discrepancy between these two planes (+/-3.75 degrees) and one with large discrepancies between these two planes (greater than 3.75 degrees or lesser than 3.75 degrees). RESULTS: Postoperative relapse was seven times more frequent when the postoperative plane tilting was superior to +/-3.75 degrees in reference to the ideal plane. DISCUSSION: Postoperative occlusal plane tilting is a predictive factor of postoperative open bite relapse. PMID- 19811795 TI - [Benign cartilaginous tumors of the jaws]. AB - Benign cartilaginous tumors are a rare entity in jaw bones. The histogenesis is still discussed but an embryological hypothesis is suggested. Chondroma, osteochondroma, chondroblastoma and chondromyxoid fibroma are the main benign maxillary cartilaginous tumors. PMID- 19811796 TI - [Reversible cerebral vasoconstriction syndrome]. AB - Reversible cerebral vasoconstriction syndrome (RCVS) is characterized by severe headaches, as well as vasoconstriction of cerebral arteries, which resolves spontaneously in one to three months. This condition has a moderate female preponderance. The mean age of onset is around 45 years. About 60% of the cases are secondary, mainly occurring during postpartum and/or after exposure to vasoactive substances. The main clinical presentation includes multiple recurrent thunderclap headaches over one to three weeks. The major complications of RCVS are localized cortical subarachnoid hemorrhages (cSAH) (20-25%) and parenchymal strokes (5-10%). Complications occur with different time courses: hemorrhages (cSAH and intracerebral hemorrhages), and posterior reversible encephalopathy syndrome are early events occurring during the first week, while ischemic events including TIAs and cerebral infarcts occur significantly later, during the second week. Diagnosis requires the demonstration of the "string and beads" aspect of cerebral arteries by a cerebral angiogram (MRA, CTA or conventional) and the demonstration of the complete or marked normalisation of arteries by a repeat angiogram performed within 12 weeks of onset. Treatment is based on nimodipine that seems to reduce thunderclap headaches within 48h. However, nimodipine has not proven any efficacy against the hemorrhagic and ischemic complications of RCVS. Relapses are possible but rare and have not been reported yet in prospective series. It seems appropriate to advise the patients to avoid sympathomimetic and serotoninergic substances. PMID- 19811797 TI - [Pharmacological hypotheses and therapeutic strategies for gait disorders in Parkinson's disease]. AB - Gait disorders form part of the axial symptoms observed in Parkinson's disease (PD) and also represent a major source of therapeutic failure in the later stages of PD, with the appearance of freezing of gait (FOG) and falls. Double-blind clinical trials and, above all, clinical experience have demonstrated that l-DOPA is effective in reducing FOG. Dopaminergic agonists appear to be less effective than l-DOPA and lack formal proof of their efficacy. The enzyme inhibitors provide modest benefits, which need to be confirmed. Hence, these symptoms appear to be partially doparesistant and justify investigation of other major neurotransmission systems. Of the various drugs with partial noradrenergic activity, methylphenidate may improve FOG and attention disorders. Memantine has shown some value in improving motor symptoms and gait in fluctuating parkinsonian patients - possibly by reducing the effect of glutamatergic hyperactivation of the subthalamic nucleus on the pedunculopontine nucleus (PPN). The PPN's dense cholinergic innervation also suggests that cholinesterase inhibitors may be of use, although any benefits must be set against a potential aggravation of rest tremor. The many interactions between the serotoninergic and dopaminergic systems require the implementation of clinical studies on the complex motor impact of serotoninergic treatments, which may aggravate the parkinsonian syndrome while improving gait (as is the case with paroxetine and ritanserin). This review seeks to develop the various pathophysiological hypotheses prompted by the results of fundamental studies and pilot clinical trials, with a view to justifying the implementation of confirmatory, double-blind, placebo-controlled therapeutic trials. PMID- 19811798 TI - [Neurosjogren: an unusual presentation with central nervous system involvement]. AB - INTRODUCTION: Neurological manifestations in Sjogren's syndrome are variable. The peripheral nervous system is generally involved. We report a rare case of an unusual central neurological manifestation. CASE REPORT: A 54-year-old woman was admitted with headache and tetraparesia. The physical examination revealed a tetrapyramidal syndrome and a bilateral parotidomegaly. The patient's general condition was nevertheless quite good. Brain MRI showed an heterogeneous pontine lesion with multiple nodular formations in the periventricular white matter. Blood tests revealed anti-SSA and anti-SSB antibodies. A labial salivary gland biopsy was grade IV in Chisholm scoring system and Schirmer's test was positive. CONCLUSION: A latent Sjogren's syndrome can lead to a wide variety of focal brain MRI abnormalities and should be evoked when the etiology is not clear. PMID- 19811799 TI - [Gait disorders - foreword]. PMID- 19811800 TI - Characterization of the colloidal organic matter from the Amazonian basin by asymmetrical flow field-flow fractionation and size exclusion chromatography. AB - Colloidal organic matter (COM) collected in small and large watercourse tributaries of the Negro River (Brazil) were fractionated and characterized by an asymmetrical flow field-flow fractionation (AFlFFF) coupled to UV and seven angle laser light scattering (LS) detectors, and by size exclusion chromatography (SEC) coupled with a UV detection. Number and weight average molar masses, weight average gyration radiuses R(Gw) and polydispersity indexes were obtained for each sample in two separate runs under conditions optimized for lower and larger size fractions. The results demonstrate the existence of a decrease of size of the colloidal matter when passing from first order streams to higher order rivers. No significant changes were found in size distributions of samples collected during the low and high flow stages at the same site. The influence of selected pre treatments such as filtration and reverse osmosis pre-concentration on the size and molar mass distributions was also studied. PMID- 19811801 TI - Simulating the dissipation of two herbicides using micro paddy lysimeters. AB - A set of packed micro paddy lysimeters, placed in a greenhouse, was used to simulate the dissipation of two herbicides, simetryn and thiobencarb, in a controlled environment. Data from a field monitoring study in 2003, including the soil condition and water balances, were used in the simulation. The herbicides were applied and monitored over a period of 21 d. The water balances under two water management scenarios, intermittent irrigation management (AI) and continuous irrigation management (CI), were simulated. In the AI scenario, the pattern of herbicide dissipation in the surface water of the field were simulated, following the first-order kinetics. In the CI scenario, similarity was observed in most lysimeter and field concentrations, but there were differences in some data points. Dissipation curves of both herbicides in the surface water of the two simulated scenarios were not significantly different (P>0.05) from the field data except for intercept of the thiobencarb curve in the CI scenario. The distribution of simetryn and thiobencarb in the soil profile after simulation were also similar to the field data. The highest concentrations of both herbicides were found on the topsoil layer at 0-2.5 cm depth. Only a small amount of herbicides moved down to the deeper soil layers. Micro paddy lysimeters are thus a good alternative for the dissipation study of pesticides in the paddy environment. PMID- 19811802 TI - Rural areas affected by the Chernobyl accident: radiation exposure and remediation strategies. AB - Main objectives of the present work were to develop an internationally agreed methodology for deriving optimized remediation strategies in rural areas that are still affected by the Chernobyl accident, and to give an overview of the radiological situation in the three affected countries, Belarus, Russia and Ukraine. Study settlements were defined by having in 2004 less than 10,000 inhabitants and official dose estimates exceeding 1 mSv. Data on population, current farming practices, contamination of soils and foodstuffs, and remedial actions previously applied were collected for each of such 541 study settlements. Calculations of the annual effective dose from internal radiation were validated with extensive data sets on whole body counter measurements. According to our calculations for 2004, in 290 of the study settlements the effective dose exceeded 1 mSv, and the collective dose in these settlements amounted to about 66 person-Sv. Six remedial actions were considered: radical improvement of grassland, application of ferrocyn to cows, feeding pigs with uncontaminated fodder before slaughter, application of mineral fertilizers for potato fields, information campaign on contaminated forest produce, and replacement of contaminated soil in populated areas by uncontaminated soil. Side effects of the remedial actions were quantified by a 'degree of acceptability'. Results are presented for two remediation strategies, namely, Strategy 1, in which the degree of acceptability was given a priority, and Remediation Strategy 2, in which remedial actions were chosen according to lowest costs per averted dose only. Results are highly country-specific varying from preference for soil replacement in populated areas in Belarus to preference for application of ferrocyn to cows in Ukraine. Remedial actions in 2010 can avert a large collective dose of about 150 person-Sv (including averted doses, which would be received in the following years). Nevertheless, the number of inhabitants in Belarusian and Russian settlements with annual doses exceeding 1 mSv remains large. Compared to international values for the cost-effectiveness of actions to reduce occupational exposures, the recommended remediation strategies for rural areas affected by the Chernobyl accident are quite cost-effective (about 20 keuro/person-Sv). PMID- 19811803 TI - Urinary phthalate monoesters and endometriosis in infertile Japanese women. AB - Phthalates may act as an estrogen and are a potential risk factor for estrogen related diseases such as endometriosis. We assessed the association between phthalate exposure and endometriosis in 166 consecutive women who presented at a university hospital for consultation regarding infertility. The subjects were interviewed and provided a urine specimen prior to a laparoscopic diagnosis of endometriosis. They were then categorized by the severity of endometriosis as controls (stages 0-I) and cases (stages II-IV). Urinary concentrations of the phthalate metabolites monoethyl phthalate, mono-n-butyl phthalate, monobenzyl phthalate, mono(2-ethylhexyl) phthalate (MEHP), mono(2-ethyl-5-oxohexyl) phthalate, and mono(2-ethyl-5-hydroxyhexyl) phthalate were measured in 57 cases and 80 controls using high-performance liquid chromatography isotope-dilution tandem mass spectrometry. Adjusted odds ratios for endometriosis in relation to dichotomized individual phthalate metabolites (standardized for creatinine) were calculated. No significant association between endometriosis and any urinary creatinine-adjusted phthalate monoester was seen. Adjusted odds ratio (95% confidence interval) for higher dichotomized MEHP by endometriosis was 1.57 (0.74 3.30). No monotonic trend was seen in urinary creatinine-adjusted concentration of phthalate metabolites by endometriosis stage (p=0.23-0.90). Our results do not support the hypothesis that higher urinary concentrations of phthalate metabolites are associated with the risk of endometriosis in infertile Japanese women. PMID- 19811804 TI - Abnormal cell-cycle expression of the proteins p27, mdm2 and cathepsin B in oral squamous-cell carcinoma infected with human papillomavirus. AB - Since the role of human papillomavirus (HPV) infection in oral carcinogenesis is still unclear, the purpose of this study was to verify the association between the expression of p27, mdm2 and cathepsin B and by HPV-related oral lesions. Fifty-five oral biopsies were studied and HPV detection and typing (6/11, 16, 18, 31 and 33) were performed using polymerase chain reaction techniques. The distribution p27, mdm2 and cathepsin B was determined by immunohistochemistry. Twenty-one (38%) out of the 55 oral lesions tested positive for HPV, of which 6 (33%) were HPV 6/11, 1 (5%) was HPV 16, 14 (72%) were HPV 18 and none was HPV 33/31. Among the 55 biopsies, immunopositivity for p27, mdm2 and cathepsin B was observed in 17 (30.9%), 37 (67.2%) and 37 (67.2%), respectively. Among 21 HPV positive oral lesions, immunopositivity of mdm2, p27 and cathepsin B was found, respectively, in 6 (33%) out of 18 benign lesions (BL), 4 (22%) out of 18 potential malignant epithelial lesions (PMEL) and 11 (57.9%) out of 19 malignant lesions (ML). High-risk HPV types may be associated with oral carcinoma, by cell cycle control dysregulation, contributing to oral carcinogenesis and the overexpression of mdm2, p27 and cathepsin B. PMID- 19811805 TI - High prevalence of extended-spectrum beta-lactamase-producing enterobacteriaceae in bacteremia after transrectal ultrasound-guided prostate biopsy: a need for changing preventive protocol. AB - OBJECTIVES: To determine whether the incidence of bacteremia after transrectal ultrasound-guided prostate biopsy (TRUSGPB) significantly diminishes with the setting up of a new preventive protocol. This protocol was set up after detecting an augmented incidence of bacteremia after TRUSGPB with a high prevalence of antibiotic-resistant microorganisms. METHODS: Retrospective descriptive and prospective intervention study performed at a University Hospital. PARTICIPANTS: Patients undergoing TRUSGPB under the old preventive protocol (January 2006 February 2007), that is, amoxicillin-clavulanate 500 mg tid the day before, the day of the procedure, and 1 day after the procedure, and after setting up a new protocol (March 2007-April 2008), that is, 2 g cefoxitin 1 hour before the procedure and ciprofloxacin 750 mg p.o. bid the day before, the day of the procedure, and 3 days after the procedure; dipstick urinalysis was performed before the procedure, and patients with positive results were not biopsied. RESULTS: Incidence of bacteremia with old and new protocols: 9 of 204 procedures (4.4%) vs 2 of 207 (0.9%), (P = .03). Four isolates (44.4%) under the old protocol produced extended-spectrum beta-lactamase (ESBL). With the new protocol, 2 (0.9%) cases of non-ESBL Escherichia coli bacteremia were observed. Sixty-five (23.8%) cases were not biopsied because of positive result of dipstick urinalysis, lack of antibiotic prophylaxis adherence, or altered coagulation parameters. CONCLUSIONS: Antibiotic prophylaxis for TRUSGPB should take into account local resistance patterns. Cefoxitin could be used as prophylaxis in centers with high prevalence of ESBL enterobacteriaceae. Before TRUSGPB, excluding patients with positive results of dipstick urinalysis is an advisable practice. PMID- 19811806 TI - Extracorporeal shock wave lithotripsy in impacted upper ureteral stones: a prospective randomized comparison between stented and non-stented techniques. AB - OBJECTIVES: To assess the need for pre-extracorporeal shock wave lithotripsy (pre ESWL) stenting in management of impacted upper ureteral stones of size < or = 2 cm and to verify whether stenting would influence the success of therapy. METHODS: Between 2007 and 2008, a total of 60 patients with solitary, radio opaque impacted upper ureteral stones measuring < or = 2 cm were divided into 2 equal groups: a stented group with a Double-J stent fixed pre-ESWL and a non stented group treated by in situ ESWL. All patients were treated by ESWL using Dornier Doli S lithotripter. Results were compared in terms of clearance rates, number of shock waves and sessions, morbidity, and incidence of complications. Pretreatment KUB (kidneys, ureters, and bladder) and intravenous pyelogram and post-treatment KUB were used to evaluate fragmentation and clearance. RESULTS: Overall stone-free rate was 88.3%. No significant statistical difference was observed in stone-free rate between the stented and non-stented groups being 90% and 86.7%, respectively (P = .346). One session was required in 28.3% of patients, whereas multiple sessions were required in 71.7% of patients. No significant statistical difference was noted in re-treatment rate in the 2 groups. Patients in the stented group significantly complained of side effects attributable to the stent predominantly dysuria, urgency, frequency, and suprapubic pain. CONCLUSIONS: ESWL is an effective and reasonable initial therapy in the management of impacted upper ureteral stones measuring < or = 2 cm. Pre ESWL ureteral stenting provides no additional benefit over in situ ESWL. Moreover, ureteral stents are associated with significant patient discomfort and morbidity. PMID- 19811808 TI - SIU Scholarship: Dr. Mohamed Zahid Musa Almasri. PMID- 19811807 TI - Trends in medical expulsive therapy use for urinary stone disease in U.S. emergency departments. AB - OBJECTIVES: Between 2000 and 2006, 11 randomized controlled trials were published, demonstrating the efficacy of medical expulsive therapy (MET) for promoting upper tract stone passage. Although its use is gaining traction among urologists, they evaluate a minority of patients who present to the emergency department (ED) for acute renal colic before discharge. As such, measuring the uptake of MET into the broader medical community is important. METHODS: Data were analyzed (2000-2006) from the National Hospital Ambulatory Medical Care Survey. Sampled ED visits for stones were identified. The use of MET was ascertained by the prescription of a calcium channel or alpha blocker at the ED visit. National estimates of the prevalence of MET use were computed. Logistic regression was used to examine linear and nonlinear time trends in MET prescription. RESULTS: The use of MET increased throughout the study period. In fact, the odds of being treated with this approach more than doubled with each successive year (OR, 2.15; 95% CI, 1.31-3.5; P < .001 for the linear trend). However, the overall prevalence of use was exceedingly low at 1.1% (95% CI, 0.6%-1.9%). Given the number needed to treat of 4, this implies a missed opportunity to spare approximately 260,000 individuals annually from stone surgery and its risks. CONCLUSIONS: Despite the growing body of evidence to support its safety and efficacy, our analysis reveals the sluggish dissemination of MET into the broader medical community. The observed underuse represents a block in the translation of clinical science into practice and raises a quality of care concern. PMID- 19811809 TI - Submission of the entire lymph node dissection for histologic examination in gynecologic-oncologic specimens. Clinical and pathologic relevance. AB - BACKGROUND AND AIMS: Lymph node (LN) status in gynecologic malignancies plays an important role in patient staging, management, and prognosis. Therefore, an adequacy of LN harvest is crucial. The aim of this study is to determine whether the submission of the entire LN dissection for histologic examination will affect patients' outcome or clinical stage. We also evaluated the time required and cost effectiveness for the laboratory. MATERIALS AND METHODS: A prospective study of 134 surgical cases from various gynecologic malignancies was conducted. The LN dissection specimen was performed using a conventional manual node dissection method with all the remaining fat being submitted in additional cassettes. One pathologist evaluated (1) the number and status of palpable LNs identified by the conventional method as well as the number of tissue cassettes and (2) the number, size, and status of the non-palpable LNs as well as the number of tissue cassettes. RESULTS: The palpable LNs ranged from 0 to 36 with average 14.8 LNs per case (Poisson 95% CI: 14.1-15.4). The additional non-palpable LNs ranged from 0 to 16 with an average of 3.1 (Poisson 95% CI: 2.8-3.4). In only one case, a 3 mm non-palpable LN with metastasis was identified; however, it did not affect tumor staging or patient management. CONCLUSION: The impact on patient outcome is minimal and it does not prove to be cost and time effective when submitting the entire LN dissection specimen in gynecologic malignancies. However, this method could be justified in selective cases in which the manual node dissection does not reveal an adequate number of LNs. PMID- 19811810 TI - Application of proteomics in ovarian cancer: which sample should be used? AB - OBJECTIVE: In the last decade several studies have been published using proteomics to unravel molecular pathways and to find biomarkers which can be used for diagnosis and/or prognostication in ovarian cancer. This review gives an overview of proteomic studies performed in ovarian cancer focusing on the nature of samples that have been used. METHODS: Recent literature regarding the role of proteomic studies in ovarian cancer has been reviewed. RESULTS: Most studies have focused on finding biomarkers for early diagnosis of ovarian cancer using blood samples though proteins identified until now are mainly acute phase reactants. Studies regarding platinum sensitivity have only been performed on cell culture models and need confirmation in tissue samples. Proteomic studies using ovarian cancer tissue are sparse and mostly contain a low number of samples. CONCLUSION: To date no biomarkers for early diagnosis or prognostication in ovarian cancer have been found using proteomics. We speculate that it would be interesting to investigate the tissue proteome in an attempt to overcome acute phase reactants and to facilitate the discovery of real tumor-specific biomarkers instead of the identification of secondary protein changes. PMID- 19811811 TI - Sunitinib malate in the treatment of recurrent or persistent uterine leiomyosarcoma: a Gynecologic Oncology Group phase II study. AB - PURPOSE: New agents are needed for patients with metastatic uterine leiomyosarcoma who progress after treatment with doxorubicin or gemcitabine docetaxel. Agents targeting tumor vasculature have potential for activity in leiomyosarcoma. We aimed to assess the activity of sunitinib in patients with recurrent uterine leiomyosarcoma who had received one or two prior therapies by determining the frequency of patients who survived progression-free for at least 6 months or who achieved objective tumor response. We also aimed to characterize the toxicity of sunitinib and to estimate time-to-progression. PATIENTS AND METHODS: Eligible patients with uterine leiomyosarcoma were treated with sunitinib 50 mg by mouth daily for 4 weeks, with 2 weeks rest. Tumor response and progression-free status were assessed every 6 weeks. RESULTS: Twenty-three of 25 patients enrolled were evaluable for efficacy (two wrong histologies). The median number of cycles was one. Two of 23 patients achieved a partial response (8.7%, 90% two-sided, binomial confidence interval (CI) 1.6-24.9%). Four patients remained progression-free at 6 months (17.4%, 90% two-sided, binomial confidence interval 6.2-35.5%). Toxicities included: grade 3 neutropenia (17.4%); grade 3 thrombocytopenia (13%); grade 3 anemia (17.4%); grades 3-4 lymphopenia (8.7%); grades 3-4 fatigue (30%); grade 3 vomiting/diarrhea (21.7%); skin rash/hand-foot syndrome, grade 2 (13%), grade 3 (4.3%); hypertension, grade 2 (39%), grade 3 (4.3%); grade 2 decrease in cardiac ejection fraction (4.3%), and grade 3 thrombosis (4.3%) Median progression-free survival (PFS) was 1.5 months. CONCLUSION: Sunitinib fails to achieve sufficient objective response or sustained disease stabilization as second- or third-line treatment for uterine leiomyosarcoma. PMID- 19811812 TI - Consistently very poorly controlled asthma, as defined by the impairment domain of the Expert Panel Report 3 guidelines, increases risk for future severe asthma exacerbations in The Epidemiology and Natural History of Asthma: Outcomes and Treatment Regimens (TENOR) study. AB - BACKGROUND: Identification of patients at risk for asthma exacerbations can assist physicians in addressing disease management and improve asthma-related health outcomes. OBJECTIVE: We sought to evaluate whether level of impairment, as defined by the 2007 asthma guidelines, predicts risk for future asthma exacerbations. METHODS: The study included children aged 6 to 11 years (n = 82) and adolescent/adult patients aged 12 years and older (n = 725) from The Epidemiology and Natural History of Asthma: Outcomes and Treatment Regimens study with data representing all components of the impairment domain of the asthma guidelines at baseline, month 12, and month 24. Patients were categorized into 2 cohorts: (1) consistently very poorly controlled (VPC) asthma from baseline through 2 years of follow-up and (2) improved from VPC asthma at baseline (including patients who improved to not well-controlled or well-controlled asthma), with improvement maintained through 2 years of follow-up. Odds ratios (ORs) and 95% CIs for risk of asthma exacerbations at month 30 were generated by using multivariable logistic regression by age group. RESULTS: After adjustment, children with consistently VPC asthma over the 2-year period demonstrated a 6 fold increased risk of hospitalization, emergency department visit, or corticosteroid burst (OR, 6.4; 95% CI, 1.2-34.5) compared with the improved group. Adolescent/adult patients with consistently VPC asthma were more likely to have a corticosteroid burst (OR, 2.8; 95% CI, 1.7-4.8) or have a hospitalization, emergency department visit, or corticosteroid burst (OR, 3.2; 95% CI, 1.9-5.3). CONCLUSIONS: Consistently VPC asthma, as defined by the impairment domain of the 2007 asthma guidelines, is strongly predictive of future asthma exacerbations. PMID- 19811813 TI - Unintended imitation in nonword repetition. AB - Verbal repetition is conventionally considered to require motor-reproduction of only the phonologically relevant content of a perceived linguistic stimulus, while imitation of incidental acoustic properties of the stimulus is not an explicit part of this task. Exemplar-based theories of speech processing, however, would predict that imitation beyond linguistic reproduction may occur in word repetition. Five experiments were conducted in which verbal audio-motor translations had to be performed under different conditions. Nonwords varying in phonemic content, in vocal pitch (F(0)), and in speaking style (schwa-syllable expression) were presented. We experimentally varied the factors response delay (repetition vs. shadowing), intention-to-repeat (repetition vs. pseudo-naming), and phonological load (repetition vs. transformation). The responses of ten healthy participants were examined for phonemic accuracy and for traces of para phonological imitation. Two aphasic patients with phonological impairments were also included, to find out if lesions to left anterior or posterior perisylvian cortex interfere with imitation. In the healthy participants, significant imitation of both F(0) and phonetic style was observed, with markedly stronger effects for the latter. Strong imitation was also found in an aphasic patient with a lesion to left anterior perisylvian cortex, whereas almost no imitation occurred in a patient with a lesion to the posterior language area. The degree of unintended imitation was modulated by each of the three independent factors introduced here. The results are discussed on the background of cognitive and neurolinguistic theories of imitation. PMID- 19811814 TI - Regional changes in word-production laterality after a naming treatment designed to produce a rightward shift in frontal activity. AB - Five nonfluent aphasia patients participated in a picture-naming treatment that used an intention manipulation (opening a box and pressing a button on a device in the box with the left hand) to initiate naming trials and was designed to re lateralize word production mechanisms from the left to the right frontal lobe. To test the underlying assumption regarding re-lateralization, patients participated in fMRI of category-member generation before and after treatment. Generally, the four patients who improved during treatment showed reduced frontal activity from pre- to post-treatment fMRI with increasing concentration of activity in the right posterior frontal lobe (motor/premotor cortex, pars opercularis), demonstrating a significant shift in lateraliity toward the right lateral frontal lobe, as predicted. Three of these four patients showed no left frontal activity by completion of treatment, indicating that right posterior lateral frontal activity supported category-member generation. Patients who improved in treatment showed no difference in lateralization of lateral frontal activity from normal controls pre-treatment, but post-treatment, their lateral frontal activity during category-member generation was significantly more right lateralized than that of controls. Patterns of activity pre- and post-treatment suggested increasing efficiency of cortical processing as a result of treatment in the four patients who improved. The one patient who did not improve during treatment showed a leftward shift in lateral frontal lateralization that was significantly different from the four patients who did improve. Neither medial frontal nor posterior perisylvian re-lateralization from immediately pre- to immediately post-treatment images was a necessary condition for significant treatment gains or shift in lateral frontal lateralization. Of the three patients who improved and in whom posterior perisylvian activity could be measured at post-treatment fMRI, all maintained equal or greater amounts of left-hemisphere perisylvian activity as compared to right. This finding is consistent with reviews suggesting both hemispheres are involved in recovery of language in aphasia patients. PMID- 19811815 TI - Responding to China's hypertensive crisis. PMID- 19811816 TI - Premature deaths attributable to blood pressure in China: a prospective cohort study. AB - BACKGROUND: Hypertension is a major global-health challenge because of its high prevalence and concomitant risks of cardiovascular disease. We estimated premature deaths attributable to increased blood pressure in China. METHODS: We did a prospective cohort study in a nationally representative sample of 169,871 Chinese adults aged 40 years and older. Blood pressure and other risk factors were measured at a baseline examination in 1991 and follow-up assessment was done in 1999-2000. Premature death was defined as mortality before age 72 years in men and 75 years in women, which were the average life expectancies in China in 2005. We calculated the numbers of total and premature deaths attributable to blood pressure using population-attributable risk, mortality, and the population size of China in 2005. FINDINGS: Hypertension and prehypertension were significantly associated with increased all-cause and cardiovascular mortality (p<0.0001). We estimated that in 2005, 2.33 million (95% CI 2.21-2.45) cardiovascular deaths were attributable to increased blood pressure in China: 2.11 million (2.03-2.20) in adults with hypertension and 0.22 million (0.19-0.25) in adults with prehypertension. Additionally, 1.27 million (1.18-1.36) premature cardiovascular deaths were attributable to raised blood pressure in China: 1.15 million (1.08 1.22) in adults with hypertension and 0.12 million (0.10-0.14) in adults with prehypertension. Most blood pressure-related deaths were caused by cerebrovascular diseases: 1.86 million (1.76-1.96) total deaths and 1.08 million (1.00-1.15) premature deaths. INTERPRETATION: Increased blood pressure is the leading preventable risk factor for premature mortality in the Chinese general population. Prevention and control of this condition should receive top public health priority in China. FUNDING: American Heart Association (USA); National Heart, Lung, and Blood Institute, National Institutes of Health (USA); Ministry of Health (China); and Ministry of Science and Technology (China). PMID- 19811817 TI - The effect of matrix stiffness on mesenchymal stem cell differentiation in a 3D thixotropic gel. AB - Recent studies have demonstrated the effect of matrix stiffness on the phenotype and differentiation pathway of mesenchymal stem cells (MSCs). MSCs differentiated into neural, myogenic or osteogenic phenotypes depending on whether they were cultured on two-dimensional (2D) substrates of elastic moduli in the lower (0.1-1 kPa), intermediate (8-17 kPa) or higher ranges (34 kPa). In this study, MSCs were cultured in thixotropic gels of varying rheological properties, and similar results were found for the three-dimensional (3D) culture as for the previous findings in 2D culture. For the 3D cell cultures in thixotropic gels, the liquefaction stress (tau(y)), the minimum shear stress required to liquefy the gel, was used to characterize the matrix stiffness. The highest expressions of neural (ENO2), myogenic (MYOG) and osteogenic (Runx2, OC) transcription factors were obtained for gels with tau(y) of 7, 25 and 75 Pa, respectively. Immobilization of the cell-adhesion peptide, RGD, promoted both proliferation and differentiation of MSCs, especially for the case of the stiffer gels (>75 Pa). This study demonstrated the usefulness of thixotropic gels for 3D cell culture studies, as well as the use of tau(y) as an effective measure of matrix stiffness that could be correlated to MSC differentiation. PMID- 19811818 TI - Concentrated collagen hydrogels as dermal substitutes. AB - Collagen hydrogels first appeared promising for skin repair. Unfortunately, their extensive contraction and their poor mechanical properties constituted major disadvantages toward their utilization as permanent graft. The present study has investigated a way to correct these drawbacks by increasing the collagen concentration in controlled conditions. Concentrated collagen hydrogels (CCH) at 1.5, 3 and 5mg/ml were obtained. The effect of raised collagen concentration on contraction, cell growth and remodeling activities was evaluated for 21 days in culture. Subsequently, in vivo integration of CCH and normal collagen hydrogels (NCH) was assessed. Compared to NCH, CCH contraction was delayed and smaller. At day 21, surface area of CCH at 3mg/ml was 18 times more important than that of NCH. Whatever the initial fibroblast density, CCH favored cell growth that reached about 10 times the initial cell number at day 21; cell proliferation was inhibited in NCH. Gelatinase A activities appeared lower in CCH than within NCH. In vivo studies in rats revealed a complete hydrolysis of NCH 15 days after implantation. In contrast, CCH at 3mg/ml was still present after 30 days. Moreover, CCH showed cell colonization, neovascularization and no severe inflammatory response. Our results demonstrate that concentrated collagen hydrogels can be considered as new candidates for dermal substitution because they are is easy to handle, do not contract drastically, favor cell growth, and can be quickly integrated in vivo. PMID- 19811819 TI - Biological performance of mussel-inspired adhesive in extrahepatic islet transplantation. AB - There is significant need for effective medical adhesives that function reliably on wet tissue surfaces with minimal inflammatory insult. To address these performance characteristics, we have generated a synthetic adhesive biomaterial inspired by the protein glues of marine mussels. In-vivo performance was interrogated in a murine model of extrahepatic syngeneic islet transplantation, as an alternative to standard portal administration. The adhesive precursor polymer consisted of a branched poly(ethylene glycol) (PEG) core, whose endgroups were derivatized with catechol, a functional group abundant in mussel adhesive proteins. Under oxidizing conditions, adhesive hydrogels formed in less than 1 min from catechol-derivatized PEG (cPEG) solutions. Upon implantation, the cPEG adhesive elicited minimal acute or chronic inflammatory response in C57BL6 mice, and maintained an intact interface with supporting tissue for up to one year. In situ cPEG adhesive formation was shown to efficiently immobilize transplanted islets at the epididymal fat pad and external liver surfaces, permitting normoglycemic recovery and graft revascularization. These findings establish the use of synthetic, biologically-inspired adhesives for islet transplantation at extrahepatic sites. PMID- 19811820 TI - Osteoblast behavior on polytetrafluoroethylene modified by long pulse, high frequency oxygen plasma immersion ion implantation. AB - Polytetrafluoroethylene (PTFE) is a commonly used medical polymer due to its biological stability and other attractive properties such as high hardness and wear resistance. However, the low surface energy and lack of functional groups to interact with the cellular environment have severely limited its applications in bone or cartilage replacements. Plasma immersion ion implantation (PIII) is a proven effective surface modification technique. However, when conducted on polymeric substrates, conventional PIII experiments typically employ a low pulsing frequency and short pulse duration in order to avoid sample overheating, charging, and plasma sheath extension. In this paper, a long pulse, high frequency O(2) PIII process is described to modify PTFE substrates by implementing a shielded grid in the PIII equipment without these aforementioned adverse effects. X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and contact angle measurements are carried out to reveal the surface effects of PTFE after long pulse, high frequency O(2) PIII and the results are compared to those obtained from conventional short pulse, low frequency O(2) PIII, O(2) plasma immersion, and the untreated control samples. Our results show that less oxygen-containing, rougher, and more hydrophobic surfaces are produced on PTFE after long pulse, high frequency O(2) PIII compared to the other 2 treatments. Cell viability assay, ALP activity test, and real-time PCR analysis are also performed to investigate the osteoblast behavior. It is clear that all 3 surface modification techniques promote osteoblast adhesion and proliferation on the PTFE substrates. Improvements on the ALP, OPN, and ON expression of the seeded osteoblasts are also obvious. However, among these treatments, only long pulse, high frequency O(2) PIII can promote the OCN expression of osteoblasts when the incubation time is 12 days. Our data unequivocally disclose that the long pulse, high frequency O(2) PIII technique is better than the other two types of traditional plasma treatment in the development of PTFE for bone or cartilage repair. PMID- 19811821 TI - Creating gradients of two proteins by differential passive adsorption onto a PEG density gradient. AB - Many fundamental biological processes, including early embryo development, immune responses and the progression of pathogens, are mediated by gradients of biological molecules. Understanding these vital physiological processes requires the development of biomaterial platforms that can mimic them in-vitro. Such platforms include laboratory generated surface gradients of biological molecules. In this work, we report a method for the generation of surface gradients of two proteins. We used a surface grafting density gradient of polyethylene glycol (PEG) to control protein adsorption. In addition, we used protein size as a tool to control the position and the adsorbed amount of both proteins. To demonstrate our concept, we used fibrinogen as an example of a large protein and lysozyme as an example of a small protein. However, we speculate that the same strategy could be extended to any other pair of large and small proteins. We used X-ray photoelectron spectroscopy and sessile drop contact angle measurements to determine the chemical composition and wettability of the gradients. Protein adsorption was studied by surface plasmon resonance imaging. PMID- 19811823 TI - Proteasome inhibitor-induced apoptosis in acute myeloid leukemia: a correlation with the proteasome status. AB - The proteasome plays a critical role in the regulation of many cellular processes, including the cell cycle and tumor growth. The proteasome inhibitor bortezomib has recently been approved for the treatment of relapsed and refractory multiple myeloma. In this study, we investigated the induction of apoptosis by proteasome inhibitors in several human acute myeloid leukemia (AML) cell lines and in primary cells from patients. We demonstrate that these drugs induce a high level of apoptosis in the KG1a cell line, in which the therapeutic drug daunorubicin is poorly active, compared to other AML cell lines. In parallel, we found that significantly different levels of apoptosis were induced in primary cells from patients depending on the FAB-based differentiation status of these cells. Moreover, the level of 20S proteasome in KG1a cells was also high compared to other AML cell lines, suggesting a relationship between the high sensitivity to proteasome inhibitors and an elevated amount of 20S proteasome. In good accordance, we identified two groups of patient cells expressing high and low levels of 20S proteasome, with respective high and low sensitivity to proteasome inhibitors. Further comparison of the proteasome status in KG1a and U937 cells also suggests that a high proportion of the 19S regulatory complex in U937 cells compared to the 20S core complex may explain an increased proteasome activity. Altogether, our results suggest that various AML subtypes may present different responses to proteasome inhibitors, that these molecules can be potentially considered as interesting therapeutic alternatives for these pathologies, and that the amount of 20S proteasome in AML cells may be predictive of the cellular response to these inhibitors. PMID- 19811822 TI - Vitamin D3-driven signals for myeloid cell differentiation--implications for differentiation therapy. AB - Primitive myeloid leukemic cell lines can be driven to differentiate to monocyte like cells by 1alpha,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), and, therefore, 1,25(OH)(2)D(3) may be useful in differentiation therapy of myeloid leukemia and myelodysplastic syndromes (MDS). Recent studies have provided important insights into the mechanism of 1,25(OH)(2)D(3)-stimulated differentiation. For myeloid progenitors to complete monocytic differentiation a complex network of intracellular signals has to be activated and/or inactivated in a precise temporal and spatial pattern. 1,25(OH)(2)D(3) achieves this change to the 'signaling landscape' by (i) direct genomic modulation of the level of expression of key regulators of cell signaling and differentiation pathways, and (ii) activation of intracellular signaling pathways. An improved understanding of the mode of action of 1,25(OH)(2)D(3) is facilitating the development of new therapeutic regimens. PMID- 19811824 TI - Major molecular response achieved with dasatinib in a CML patient with F317L BCR ABL kinase domain mutation. PMID- 19811825 TI - Isolated CNS relapse of CML after bone marrow transplantation. PMID- 19811826 TI - Child maltreatment and adult socioeconomic well-being. AB - OBJECTIVE: Little empirical research has examined the impact that child maltreatment may have on victims' long-term socioeconomic well-being. The current study sought to address this gap by exploring the relationship between childhood experiences of abuse and neglect and several indicators of socioeconomic well being in adulthood. METHOD: Data from the nationally representative National Comorbidity Survey (NCS) (n=5004) were analyzed using logistic regression models to examine whether maltreatment in childhood (any maltreatment, physical abuse, sexual abuse, severe neglect, and multiple types of maltreatment) affected employment status, income, and health care coverage in adulthood. Several potential confounds of this relationship were included as covariates in the models, including race, sex, age, and several indicators of childhood socioeconomic status (SES). RESULTS: The results show that adults who had experienced maltreatment differed significantly from non-maltreated adults across each of the socioeconomic domains examined. Effects were additionally found to differ depending on the number of types of maltreatment experienced. CONCLUSIONS: Increased rates of unemployment, poverty, and Medicaid usage indicate the significant long-term personal impact of early victimization. They also suggest a substantial societal cost from this problem through lost economic productivity and tax revenue, and increased social spending. Low socioeconomic status among parents has also been identified as a salient risk factor for the perpetration of maltreatment, and, as such, these results indicate a potential mechanism in the intergenerational transmission of violence. PRACTICE IMPLICATIONS: The findings from this study suggest that victims of child maltreatment are at increased risk for financial and employment-related difficulties in adulthood. Approximately one million children are identified each year by state agencies as victims of maltreatment in the United States. Many maltreated children, furthermore, go undetected by protective service agencies, indicating the high prevalence of this problem, and underscoring its large economic costs to society. By highlighting the long-term socioeconomic costs of maltreatment, this research should encourage policy makers to focus on improving prevention, intervention, and treatment efforts for victims of abuse and neglect. PMID- 19811828 TI - Disparities in the medical examination of children in the home of a child with suspected physical abuse. PMID- 19811827 TI - Men's and women's childhood sexual abuse and victimization in adult partner relationships: a study of risk factors. AB - OBJECTIVES: (1) Document the prevalence of childhood sexual abuse (CSA), childhood physical assault, psychological, physical and sexual intimate partner violence (IPV) in a nationally representative sample. (2) Assess the predictive value of CSA and other characteristics of the respondents and their current partners as potential risk factors for IPV. (3) Assess factors predicting IPV in adulthood in a subsample of women reporting CSA. METHODS: The role of CSA as a risk factor for adult IPV was examined using data from the 1999 Canadian General Social Survey. A national stratified sample of 9170 women and 7823 men with current or previous partners were interviewed by telephone by Statistics Canada. Multiple logistic regressions were used. RESULTS: CSA consistently predicted IPV for women and men, although this relationship was weaker for men. Age, current marital status and limitations due to physical or mental condition or chronic illness were also predictors of IPV for men and women. For women reporting CSA, age (being younger) or being in a more recent relationship and being limited due to either physical, mental conditions or chronic illness were predictive of adult victimization. CONCLUSIONS: These findings indicate that CSA is associated with a greater risk of IPV beyond sociodemographic risk factors. PRACTICE IMPLICATIONS: To prevent IPV in women already at risk because of CSA, education about protective strategies seems important, particularly for women with physical or mental limitations, in the beginning stages of intimate relationships or for women with partners who drink excessively. PMID- 19811829 TI - Tourniquet syndrome: interest of a systematic analysis of families' social conditions to detect neglect situations. PMID- 19811830 TI - The mental and physical health difficulties of children held within a British immigration detention center: a pilot study. AB - OBJECTIVE: The present study aimed to assess the mental and physical health of children held within a British immigration detention center. METHOD: A total of 24 detained children (aged 3 months to 17 years) were assessed with their parents or carer after being referred by a registered legal charity. Thirteen were seen by a pediatrician alone, 4 by a psychologist alone, and 7 by both professions using semi-structured clinical interviews. The psychologist also used standardized self-report questionnaires to measure psychopathology. RESULTS: During the psychological assessment of 11 children, 8 met criteria for psychiatric "caseness" on the Strengths and Difficulties Questionnaire. All 11 reported symptoms of depression and anxiety. Sleep problems, somatic complaints, poor appetite, emotional symptoms, and behavioral difficulties were common. Symptoms of global distress were also reported by all 9 parents. According to pediatric assessment 8 out of 20 children had lost weight. Six had missed health appointments and 2 were taken to hospital. Nutritional, developmental, educational, and child protection concerns were raised. CONCLUSIONS: Detained children were found to be experiencing mental and physical health difficulties of recent onset, which appeared to be related to the detention experience. These findings support previous Australian studies demonstrating that detention is not in the best interest of the child. It suggests that current UK policies regarding the detention of children for purposes of immigration control should be re examined. Further research in the area is required. PRACTICE IMPLICATIONS: Although high levels of mental and physical health problems, as well as child protection concerns were detected, detained families had very limited access to appropriate assessment, support or treatment. The traumatic experience of detention itself also has implications for the sizeable proportion of psychologically distressed children who are eventually released from detention and expected to successfully reintegrate into British society; while those children who are deported are returned with increased vulnerability to future stressors. PMID- 19811832 TI - Genotoxicity of two pathogenic strains of zoosporic fungi (Achlya klebsiana and Aphanomyces laevis) on erythrocytes of Nile tilapia Oreochromis niloticus niloticus. AB - In the present work we have described the genotoxic potential of two pathogenic strains of zoosporic fungi (Achlya klebsiana and Aphanomyces laevis) on erythrocytes of Nile tilapia Oreochromis niloticus niloticus for the first time by three complementary tests: micronucleus test (MN), nuclear lesions (NL) and comet assay (CA). The groups exposed to the zoosporic fungi subjected to the MN and NL test showed statistically significant differences in MN and NL frequencies with respect to the control one. Also, a significant increase (p<0.001) in micronuclei and nuclear lesions frequencies were recorded with the increase in exposure time. A correlation was observed between the frequencies of MN and NL, suggesting the importance for recording this anomaly in order to improve the information obtained with the MN test. Therefore, our results suggest that the nuclear lesions found here should be considered indicators of genotoxicity, in addition to the typical micronuclei forms. The result of the comet assay showed a significant difference in the percentages of the damaged DNA in the comet tail (%TDNA) of the treated groups comparing to the control. A significant (p<0.001) increase in the DNA damage of the treated groups with the increase in exposure time was recorded, confirming the results of the MN and the NL tests. The results of the current study will be useful for future work involving the biomonitoring of regions where Nile tilapia survive. These data allow us to consider O. niloticus niloticus as a good bioindicator of the effects of genotoxic agents that might be present in the aquatic habitat. PMID- 19811831 TI - Tolerance of arsenate-induced stress in Aspergillus niger, a possible candidate for bioremediation. AB - The arsenate tolerance limit in wild-type Aspergillus niger was determined. Because of its high tolerance, toxic effects of arsenate concentrations ranging from 25 to 100mg/L were investigated in regard to growth, intracellular thiols, proline and malondialdehyde (MDA) contents of wild-type A. niger. Cellular arsenate uptake was analyzed. Activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and succinate dehydrogenase (SDH) were assayed. Growth of A. niger increased at 25mg/L arsenate, and it survived up to 100mg/L. MDA, intracellular thiol and proline contents increased up to a certain level. Activities of GR, SOD and CAT declined following a rise at low concentration(s); SDH activity decreased gradually with increased arsenate stress. Results indicated that A. niger had high arsenate uptake potential and could tolerate oxidative stress by manipulating its anti-oxidative defense mechanism, a property that may be exploited for removal of arsenate from contaminated aqua-environment. PMID- 19811833 TI - Effects of ammonium on the antioxidative response in Hydrilla verticillata (L.f.) Royle plants. AB - To investigate ammonium toxicity, the submerged plant Hydrilla verticillata (L.f.) Royle was treated with 0.1-3.0mM ammonium for 12h and 4d. After exposure to ammonium for 4d, content of O2(-) and H2O2 increased in leaves of H. verticillata exposed to 3mM ammonium compared with control (0mM NH4Cl), while the malondialdehyde content decreased. The chlorophyll (a+b) and carotenoid concentrations decreased in H. verticillata plants exposed to 1.5-3mM ammonium for 12h and 4d. Compared with controls, the activity of superoxide dismutase, peroxidase, catalase, glutathione reductase, ascorbate peroxidase, and glutathione S-transferase increased in plants treated with ammonium for 12h, and the activity of most enzymes was further enhanced at 4d. The changes in nonprotein thiols, total glutathione, ascorbic acid, and dehydroascorbate content were also assayed. Our results suggest that ammonium induced the oxidative stress and the heated antioxidant response in H. verticillata. PMID- 19811834 TI - Immunomodulation of caprine lentiviral infection by interleukin-16. AB - Interleukin-16 (IL-16) is a proinflammatory cytokine produced by a variety of cells including lymphocytes, macrophages, mast cells, and eosinophils. We have shown in our previous studies increased expression of IL-16 mRNA and protein in caprine arthritis-encephalitis virus (CAEV)-infected goats blood. In this study, we determined the immunomodulatory effects of IL-16 in vitro using cells derived from CAEV infected and uninfected goats. Human recombinant IL-16 (rhIL-16) significantly increased chemotaxis of peripheral blood mononuclear cells (PBMCs) of both control and CAEV-infected goats. Pretreatment of PBMC with anti-goat CD4 monoclonal antibody inhibited IL-16-induced chemotaxis of PBMC of control and infected goats suggesting that IL-16 exerts its action in goats primarily by binding to CD4. The CAEV proviral DNA was less in caprine monocytes treated with rhIL-16 infected in vitro with CAEV. These data suggest inhibitory effect of IL 16 on viral integration. Flow cytometric studies indicated a trend toward IL-16 induced increased expression of lymphocyte activation markers. Combined with our previously reported data, these experiments suggest that increased IL-16 expression during CAEV infection may inhibit viral integration. PMID- 19811835 TI - Accountability and psychiatric disorders: how do forensic psychiatric professionals think? AB - Swedish penal law does not exculpate on the grounds of diminished accountability; persons judged to suffer from severe mental disorder are sentenced to forensic psychiatric care instead of prison. Re-introduction of accountability as a condition for legal responsibility has been advocated, not least by forensic psychiatric professionals. To investigate how professionals in forensic psychiatry would assess degree of accountability based on psychiatric diagnoses and case vignettes, 30 psychiatrists, 30 psychologists, 45 nurses, and 45 ward attendants from five forensic psychiatric clinics were interviewed. They were asked (i) to judge to which degree (on a dimensional scale from 1 to 5) each of 12 psychiatric diagnoses might affect accountability, (ii) to assess accountability from five case vignettes, and (iii) to list further factors they regarded as relevant for their assessment of accountability. All informants accepted to provide a dimensional assessment of accountability on this basis and consistently found most types of mental disorders to reduce accountability, especially psychotic disorders and dementia. Other factors thought to be relevant were substance abuse, social network, personality traits, social stress, and level of education. PMID- 19811836 TI - Kinetics and thermodynamics of interaction of coagulation factor VIII with a pathogenic human antibody. AB - Replacement therapy in hemophilia A with exogenous coagulation factor VIII (FVIII) often results in the development of FVIII-neutralizing antibodies, referred to as inhibitors. Despite of large number of studies on the functional properties of FVIII inhibitors, detailed physicochemical characterization of their interactions is not available. Here we studied the biophysical mechanism of the interaction between a human pathogenic antibody--BO2C11 and its target antigen--FVIII. Kinetic and thermodynamic analyses implied that this interaction is not accompanied by significant conformational changes in the proteins. The data also suggested that association of BO2C11 to FVIII is driven mainly by a hydrophobic effect. The protein electrostatics however played a decisive role in this association. Thus, a gradual increase in ionic strength resulted in a considerable increase in the association rate of binding of BO2C11 to FVIII. Such an ionic strength-dependency is uncommon for other antibody-antigen interactions. Our data suggest that electrostatic effects observed for BO2C11-FVIII association may arise from high-energy penalty of desolvation of the charged residues at the binding interfaces. We hypothesize that untypical ionic strength dependence of association of BO2C11 to FVIII reflects the nature of the recognized epitope, namely a molecular surface involved in the binding of FVIII to phospholipids. The presented data provide mechanistic information about FVIII neutralization by an inhibitory antibody and also contribute to the understanding of the general mechanisms of antibody-antigen interactions. PMID- 19811837 TI - Complement receptor 3, not Dectin-1, is the major receptor on human neutrophils for beta-glucan-bearing particles. AB - We investigated the role of the beta-glucan receptor, Dectin-1, in the response of human neutrophils to unopsonized Saccharomyces cerevisiae and its major beta glucan-containing capsular constituent, zymosan. Although reported to be indispensable for yeast phagocytosis in murine phagocytes, human Dectin-1 was not involved in the phagocytosis of S. cerevisiae or zymosan by human neutrophils. Phagocytosis of yeast particles proved to be completely dependent on CD11b/CD18, also known as complement receptor 3 (CR3). The findings were supported by data with neutrophils from a patient suffering from Leukocyte-Adhesion Deficiency type 1 (LAD-1) syndrome lacking CD11b/CD18. In addition, neither the priming by zymosan of the fMLP-induced NADPH-oxidase activity in human neutrophils nor the secretion of IL-8 by human neutrophils in response to zymosan preparations was affected by blocking anti-Dectin-1 antibodies or laminarin as a monovalent inhibitor. As shown by neutrophils from an IRAK-4-deficient patient, the zymosan induced IL-8 release was also independent of TLR2. In summary, our data show that Dectin-1, although indispensable for recognition of beta-glucan-bearing particles in mice, is not the major receptor for yeast particles in human neutrophils. PMID- 19811838 TI - Obsessive-compulsive behavior spectrum: refining the research agenda for DSM-V. AB - The American Psychiatric Association, in collaboration with the World Health Organization and the National Institutes of Health, has undertaken a 5-year international research planning effort in preparation for the formal revision of the Diagnostic and Statistical Manual of Mental Disorders. One element of the project was a conference titled "Obsessive-Compulsive Behavior Spectrum," in which participants reviewed an array of disorders that cross current diagnostic categories. Questions raised challenge those responsible for the DSM-V revision to assess the pros and cons of changing definitions, boundaries, or linkages among diverse conditions characterized by obsessive-compulsive behaviors in the revised classification. PMID- 19811839 TI - Cross-cutting issues and future directions for the OCD spectrum. AB - The research planning agenda for DSM-V examined possible similarities in phenomenology, comorbidity, familial and genetic features, brain circuitry, and treatment response between obsessive-compulsive disorder (OCD) and several related disorders that are characterized by repetitive thoughts or behaviors. Such data support a re-examination of the DSM-IV-TR classification of OCD and the anxiety disorders, with possible inclusion of a group of obsessive-compulsive spectrum disorders (OCSDs) in DSM-V. Various disorders were systematically examined for inclusion in such a grouping, and later a smaller number were determined to meet threshold criteria for inclusion in the OCSDs. The disorders that were originally examined included OCD, obsessive-compulsive personality disorder (OCPD), Tourette's syndrome (TS) and other tic disorders, Sydenham's chorea, Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcal Infections (PANDAS), trichotillomania (TTM), body dysmorphic disorder (BDD), autism, eating disorders, Huntington's and Parkinson's disease, impulse control disorders, as well as substance and behavioral addictions. Certain disorders such as BDD, OCPD, TS, and TTM share many commonalities with OCD in phenomenology, comorbidity, familial and genetic features, brain circuitry, and treatment response. Other disorders, such as the impulse control disorders (ICDs) share some common features with OCD, but also differ in many ways as well. The articles presented in this issue of Psychiatry Research are a result of this international collaboration, which examined diagnostic and classification issues of OCSDs for DSM-V in a conference titled "The Future of Psychiatric Diagnosis: Refining the Research Agenda: Obsessive-Compulsive Behavior Spectrum" held in June 2006 at the American Psychiatric Association's headquarters in Arlington, VA. PMID- 19811841 TI - Gene expression profiling and environmental contaminant assessment of migrating Pacific salmon in the Fraser River watershed of British Columbia. AB - The health and physiological condition of anadromous salmon is of concern as their upriver migration requires navigation of human-impacted waterways and metabolism of stored energy reserves containing anthropogenic contaminants. Such factors may affect reproductive success of fish stocks. This study investigates chemical contaminant burdens and select gene expression profiles in Pacific Sockeye (Oncorhynchus nerka) and Chinook (Oncorhynchus tshawytscha) salmon which traverse the Fraser River watershed during their spawning migration. Chemical analyses of muscle tissue and eggs of salmon collected from the lower Fraser River (pre-migration) and from upstream spawning grounds (post-migration) during the 2007 migration revealed the presence of numerous chemical contaminants, including PCBs, dioxins/furans, pesticides, and heavy metals. However, muscle tissue residue concentrations were well below human health consumption guidelines and 2,3,7,8 TCDD toxic equivalents (SigmaTEQs) in salmon eggs, calculated using WHO toxic equivalency factors (WHO-TEFs) for fish health, did not exceed the 0.3pgg(-1) wet weight toxicological threshold level previously associated with 30% egg mortality in salmon populations. Quantitative real-time PCR probes were generated and used to assess differences in abundance of key mRNA transcripts encoding nine gene products associated with reproduction, stress, metal toxicity, and exposure to environmental contaminants. Gene expression profiles were characterized in liver and muscle tissue of pre- and post-migration Sockeye and Chinook salmon. The results of stock-matched animals indicate that dynamic changes in mRNA levels occur for a number of genes in both species during migration and suggest that Sockeye salmon exhibit a greater level of biological stress compared to the Chinook salmon population. Using a male-specific genotypic marker, we found that out of the 154 animals examined, one Sockeye was genotypically male but phenotypically female. This individual's gene expression profile in liver and muscle was reminiscent of, but not identical to, the female expression profile. These studies provide the first glimpse of the dynamic yet common nature of changes in the transcriptome that are shared between species during in-migration and highlight differences that may relate to population success. Continued longitudinal assessment will further define the association between contaminant burden, physiological stress, and modulation of gene expression in migrating Pacific salmon. PMID- 19811842 TI - Assessment of thyroid system disruption in Rana pipiens tadpoles chronically exposed to UVB radiation and 4-tert-octylphenol. AB - Many studies have considered recent increases in ultraviolet B radiation (UVBR) and endocrine disrupting chemicals polluting the environment as possible contributing factors to the reduction in amphibian populations. It has been demonstrated that exposure of amphibians to estrogenic chemicals or UVBR can affect the timing of larval development and metamorphosis. However, amphibians in the wild are exposed to multiple environmental stressors simultaneously. Therefore, our study examines the effects of UVBR and the estrogenic chemical 4 tert-octylphenol (OP), alone and in combination, on the thyroid system of Rana pipiens tadpoles, which is the main regulator of amphibian metamorphosis. Results demonstrate that thyroid gland histomorphology measurements in Gosner stage 31 tadpoles continuously exposed to UVBR (0.21W/m(2)) were not different than those measured in animals from the control group. In a separate experiment, tadpoles exposed to environmentally relevant levels of UVBR (0.22W/m(2)) and/or OP (0.01nM or 10nM) exhibited significantly delayed development starting from Gosner stage 29, given that fewer tadpoles developed past stage 29 in these groups. In addition, significantly fewer UVBR-treated tadpoles developed past stage 34 and metamorphosed. Samples were collected from stages 29 and 34 tadpoles for gene expression analysis in tail tissue and measurements of T3 (triiodothyronine) whole body levels (minus tail). UVBR and/or OP exposure did not affect T3 levels in stages 29 and 34 tadpoles. However, a decrease in deiodinase type 2 (D2) or increase in deiodinase type 3 (D3) mRNA levels was observed in groups of tadpoles with slowed developmental rates at those developmental stages. Given that D2 activates and D3 inactivates thyroid hormones (TH), UVBR/OP mediated disruptions in development are likely caused by dysfunctions in the localized metabolism of THs through alterations in the expression of these enzymes in peripheral tissues. This is the first study to our knowledge reporting a potential thyroid-based mechanism of action for the developmental delays in amphibians exposed to UVBR and/or OP. PMID- 19811840 TI - The relationship between impulse-control disorders and obsessive-compulsive disorder: a current understanding and future research directions. AB - Impulse-control disorders (ICDs) constitute a heterogeneous group of conditions linked diagnostically by difficulties in resisting "the impulse, drive, or temptation to perform an act that is harmful to the person or to others." Specific ICDs share clinical, phenomenological and biological features with obsessive-compulsive disorder (OCD) that have suggested that these disorders might be categorized together. However, other data suggest significant differences between OCD and ICDs. In this article, clinical, phenomenological and biological features of the formal ICDs are reviewed and compared and contrasted with those of OCD. Available data indicate substantial differences between ICDs and OCD that suggest independent categorizations. Existing research gaps are identified and avenues for future research suggested. PMID- 19811843 TI - A stochastic model for the assessment of the transmission pathways of heart and skeleton muscle inflammation, pancreas disease and infectious salmon anaemia in marine fish farms in Norway. AB - Salmon farming is threatened, economically and ecologically, by infectious diseases. To reduce the risk of epidemics, authorities have developed regulations. These are based on quantitative understanding of pathways of infection, representing disease specific risks. A stochastic model was fitted to historical data, to estimate risk factors associated with competing spread mechanisms. Three infectious diseases were compared, heart and skeletal muscle inflammation (HSMI), pancreas disease (PD) and infectious salmon anaemia (ISA). This study was based on space-time data, from Norway from 2003 to 2007, describing the susceptible fish cohorts and the reported infections. Particular interest was given to seaway distances between farms and their local management organisation. The parameter measuring the effect of distance to an infectious fish farm was positive and significant for all diseases, implying that the risk involved with proximate infectious fish farms increased with decreasing distance. For HSMI and PD there was a significant effect of sharing a contact network with an infectious farm. For HSMI, but not for PD or ISA, there was a significant effect of previous infected cohorts on the same farm. The relative contribution of each transmission pathway was dominated by seaway distance for PD and HSMI, while other non-defined pathways dominated for ISA. This comparative study highlights that the three diseases have different patterns of spread, with important consequences for disease prevention and management. PMID- 19811844 TI - Special section: Schwabe Symposium 2008. Preface. PMID- 19811845 TI - Education of global veterinarians. AB - In 2003, the University of Florida (UF) College of Veterinary Medicine (CVM) created an Office of International Programs (OIP) in response to one of ten initiatives of the UF Strategic Plan: internationalization of the curriculum. The OIP has developed coursework that provides students with an opportunity for international exposure during the veterinary curriculum at three levels. In Level 1 (on campus) students can participate in a seminar series in global health: www.ufglobalhealth.org. This is an elective course offered to professional students at the UF Health Science Center (Dentistry, Medicine, Pharmacy, Public Health, and Veterinary Medicine). In Level 2 (abroad), students can participate in structured study abroad programs under the supervision of UF faculty and international scholars from collaborative institutions abroad. In Level 3 (on campus and abroad), students can participate in a certificate program in international veterinary medicine. This is a 15-credit program, parallel to the veterinary curriculum. By offering courses on campus and abroad, we want to empower the curriculum with a global perspective of the veterinary profession, as well as with a humanist education that can help students recognize the importance of respect for cultural differences and the reasons for different degrees of development and growth in the world. In addition, this paper presents the need for veterinary medicine and other disciplines in the health sciences to communicate with other disciplines in the social sciences and natural sciences to create development practitioners equipped with cross-disciplinary knowledge and skills needed to formulate, implement and evaluate solutions aimed at breaking the cycle of poverty and disease in low income societies. Finally, this paper makes a call to the American Veterinary Medical Association Council on Education to assess the need to recognize the importance of internationalization of the veterinary curriculum as a key standard for accreditation of colleges or schools of veterinary medicine. PMID- 19811846 TI - Thermobacteriological characterization of Enterobacter sakazakii. AB - In the present study the influence of various environmental and physiological factors on the heat resistance of Enterobacter sakazakii (Cronobacter) have been investigated. Our results demonstrated that the heat resistance of E. sakazakii depended on the strain studied, the growth conditions - phase and temperature - the characteristics of treatment medium and the recovery conditions. The strain STCC 858 (ATCC type strain 29544) showed maximum heat resistance among the strains tested and it was selected for the further study. Stationary-phase cells grown between 20 and 37 degrees C (mean D(60)=0.9 min) resulted to be more resistant than cells grown at 10 degrees C (D(60)=0.2 min). Resistance decreased when the treatment medium pH was lower than pH 6.0, and it increased with decreasing water activity of the treatment medium, with a 32-fold increase in resistance when lowering water activity to 0.96. z value at pH 4.0 (z=4.79 degrees C) was significantly higher than at pH 7.0 (z=4.06 degrees C), although E. sakazakii cells were approximately 10 times more heat resistant at pH 7.0 than at pH 4.0 within the range of temperatures tested. Contrary to pH, the magnitude of the influence of a(w) on heat resistance did not significantly change with treatment temperature. The proportion of sublethally damaged cells was similar regardless of the treatment medium pH, but it decreased when lowering the water activity. Nevertheless, increasing treatment temperature would not result in a decreased proportion of sublethally injured E. sakazakii cells within the surviving population. Thus, the design of a theoretical combined process that could take advantage of the occurrence of sublethally injured cells would be similarly effective at low and high temperatures. E. sakazakii proved to be more heat resistant in four different liquid food matrixes than in buffers at the same pH, and this disagreement was especially higher in orange juice, which resulted to be the product that induced a greater protective effect in E. sakazakii cells against heat. PMID- 19811847 TI - Early insulin response and insulin sensitivity are equally important as predictors of glucose tolerance after correction for measurement errors. AB - AIMS: We estimated measurement error (ME) corrected effects of insulin sensitivity (M/I), from euglycaemic insulin clamp, and insulin secretion, measured as early insulin response (EIR) from oral glucose tolerance test (OGTT), on fasting plasma glucose, HbA1c and type 2 diabetes longitudinally and cross sectional. METHODS: In a population-based study (n=1128 men) 17 men made replicate measurements to estimate ME at age 71 years. Effect of 1 SD decrease of predictors M/I and EIR on longitudinal response variables fasting plasma glucose (FPG) and HbA1c at follow-ups up to 11 years, were estimated using uncorrected and ME-corrected (with the regression calibration method) regression models. RESULTS: Uncorrected effect on FPG at age 77 years was larger for M/I than for EIR (effect difference 0.10mmol/l, 95% CI 0.00;0.21), while ME-corrected effects were similar (0.02mmol/l, 95% CI -0.13;0.15mmol/l). EIR had greater ME-corrected impact than M/I on HbA1c at age 82 years (-0.11%, -0.28; -0.01%). CONCLUSIONS: Due to higher ME effect of EIR on glycaemia is underestimated as compared with M/I. By correcting for ME valid estimates of relative contributions of insulin secretion and insulin sensitivity on glycaemia are obtained. PMID- 19811848 TI - Treatment of psoriasis in patients with hepatitis C: from the Medical Board of the National Psoriasis Foundation. AB - BACKGROUND: Treating psoriasis in patients with concomitant hepatitis C virus (HCV) infection presents a special challenge. Not only is psoriasis exacerbated by interferon therapy, the standard of care for HCV, but many psoriasis therapies are potentially hepatotoxic, immunosuppressive, or both, which has been generally thought to be a contraindication in chronic infections such as HCV. OBJECTIVE: Our aim was to arrive at a consensus on treating psoriasis in patients with concomitant HCV infection. METHODS: Reports in the literature were reviewed regarding common psoriasis therapies and liver toxicity. RESULTS: Topical therapies are first-line therapy for patients with limited psoriasis and HCV. Ultraviolet B phototherapy may be considered as a second-line treatment when needed. Ultraviolet B phototherapies in combination with topical therapies are first line for patients with moderate to severe psoriasis, and are considered safe in those patients with concomitant HCV infection. Other systemic therapies, such as acitretin, etanercept, and, possibly, other tumor necrosis factor inhibitors, are considered second line. Psoralen plus ultraviolet A should also be considered a second-line therapy. LIMITATIONS: There are few evidence-based studies on treating psoriasis with systemic therapy in patients with pre-existing liver disease. CONCLUSIONS: There are no large double-blind clinical trials addressing the treatment of psoriasis in patients with HCV infection and more studies are needed. PMID- 19811849 TI - Cutaneous B-cell lymphoma with histologic features of mycosis fungoides. AB - Histologic examination of skin biopsy specimens from a 57-year-old man with a pruritic rash on his chest, abdomen, and thighs revealed a dense atypical dermal lymphoid infiltrate bordering the dermoepidermal junction and scattered intraepithelial lymphocytes. Histopathologic and clinical features were suggestive of mycosis fungoides. Immunophenotyping studies, however, identified CD20(+) B lymphocytes with aberrant expression of CD43. Clonal immunoglobulin gene rearrangement was demonstrated, and no clonal T-cell gene rearrangement was identified. Morphologic and immunophenotypic features were most consistent with extranodal marginal zone B-cell lymphoma. Systemic evaluation identified involvement of the bone marrow and possibly the peripheral blood, spleen, and splenic lymph nodes. Cutaneous manifestations were treated with narrowband ultraviolet B phototherapy for 3 months, resulting in complete resolution of the pruritus and rash. Primary and secondary cutaneous B-cell lymphomas seldom mimic mycosis fungoides histologically. Immunophenotyping studies were critical in correctly classifying this rare example of an epidermotropic B-cell lymphoma. PMID- 19811850 TI - Guidelines of care for the management of psoriasis and psoriatic arthritis: Section 5. Guidelines of care for the treatment of psoriasis with phototherapy and photochemotherapy. AB - Psoriasis is a common, chronic, inflammatory, multisystem disease with predominantly skin and joint manifestations affecting approximately 2% of the population. In this fifth of 6 sections of the guidelines of care for psoriasis, we discuss the use of ultraviolet (UV) light therapy for the treatment of patients with psoriasis. Treatment should be tailored to meet individual patients' needs. We will discuss in detail the efficacy and safety as well as offer recommendations for the use of phototherapy, including narrowband and broadband UVB and photochemotherapy using psoralen plus UVA, alone and in combination with topical and systemic agents. We will also discuss the available data for the use of the excimer laser in the targeted treatment of psoriasis. Finally, where available, we will summarize the available data that compare the safety and efficacy of the different forms of UV light therapy. PMID- 19811851 TI - Stevens-Johnson syndrome and toxic epidermal necrolysis in Asian children. AB - BACKGROUND: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are rare but severe drug reactions. There have been few reviews of SJS and TEN in children. OBJECTIVES: To evaluate the clinical profile and treatment outcomes of 15 pediatric patients with SJS or TEN. METHODS: We retrospectively reviewed the case notes of all patients diagnosed with SJS or TEN admitted to a tertiary care pediatric hospital from 2001 to 2006. RESULTS: We identified 13 cases of SJS, 1 case of SJS/TEN overlap and 1 case of TEN. Four patients were treated with intravenous immunoglobulin (IVIg), 5 patients were treated with systemic corticosteroids, and 6 patients were treated with supportive therapy only. The time to cessation of progression of disease was not significantly different in these 3 groups of patients. The duration of hospital stay was longer for patients treated with IVIG compared with those treated with systemic corticosteroids or supportive therapy. The only death was the patient with TEN treated with IVIG. LIMITATIONS: This was a retrospective study with a very small number of patients. CONCLUSION: The use of intravenous immunoglobulins or systemic corticosteroids did not improve the outcome of SJS and TEN. PMID- 19811852 TI - The changing profile of patients who used emergency department services in the United States: 1996 to 2005. AB - STUDY OBJECTIVE: Because of the vital role of emergency departments (EDs) in the US health care system, it is important to monitor the changes in the patient mix over time to identify existing problems and ways to improve the system. The current study aimed to identify raw and population-adjusted time trends for demographic characteristics, socioeconomic characteristics, access to care, utilization of care, and general health of ED users and heavy ED users. METHODS: Ten years' worth of nationally representative data was derived from the Medical Expenditure Panel Survey, 1996 to 2005. Raw time trends of various patient characteristics for ED users, nonusers, and heavy users were estimated to demonstrate changes in ED patient mix. Population-attributable fraction was used to generate population-adjusted trends that elucidate the net changes in subpopulations' propensities to ED use in relation to those of the general population. RESULTS: Between 1996 and 2005, the total number of noninstitutionalized individuals who used ED services increased from 34.2 to 40.8 million. That is, the proportion of ED users in the US population increased from 12.7% to 13.8%. Increasing proportions of elderly and those who perceived themselves to be in poor or fair physical health among ED users, particularly heavy users, were found in both raw and population-adjusted trends. Several subpopulations demonstrated increasing levels of ED use after population adjustment: blacks, patients within 100% to 199% of the federal poverty line, patients with only Medicare, patients with greater than or equal to 2 types of insurance, and patients with at least 1 inpatient stay. Decreasing population adjusted trends were found in the proportions of female patients, Hispanics, patients at greater than or equal to 200% of the federal poverty line, the uninsured, and patients with only private insurance, respectively. CONCLUSION: EDs play a larger role in the management of geriatric patients over time. The increasing burden of the aging population in the EDs poses challenges in the training of future emergency physicians, care for older patients, public health insurance, and health care system reform. PMID- 19811854 TI - [Patients over 80 years in intensive medicine]. PMID- 19811853 TI - [Diagnostic and therapeutic alternatives in perioperative acute myocardial ischemia in heart surgery]. AB - Ischemia and infarct after surgical revascularization are a relatively frequent complication, with high morbidity and mortality. Early diagnosis is essential. However, this is less standardized and more complicated to diagnose than in patients who have not undergone surgery since there is no specific biomarker that allows the clinician to differentiate between myocardial ischemia due to the procedure itself and myocardial damage due to perioperative infarct. Once detected, perioperative ischemia should be treated immediately in order to limit myocardial damage. The objectives of this study have been 1. To show the diagnostic criteria for perioperative infarct and ischemia. 2. to show the different therapeutic options available. 3. to propose a treatment algorithm that includes the differential diagnosis, how to control vasospasm, implantation of balloon counterpulsation, and the possible revascularization strategies (percutaneous coronary intervention vs reoperation). PMID- 19811855 TI - [Telemedicine: Improving the quality of care for critical patients from the pre hospital phase to the intensive care unit]. AB - The Health System is in crisis and critical care (from transport systems to the ICU) cannot escape from that. Lack of integration between ambulances and reference Hospitals, a deep shortage of critical care specialists and assigned economical resources that increase less than critical care demand are the cornerstones of the problem. Moreover, the analysis of the situation anticipated that the problem will be worse in the future. "Closed" ICUs in which critical care specialists direct patient care outperform "open" ones in which primary admitting physicians direct patient care in consultation with critical care specialists. However, the current paradigm in which a critical care specialist is close to the patient is in the edge of the trouble so, only a new paradigm could help to increase the number of patients under intensivist care. Current information technology and networking capabilities should be fully exploited to improve both the extent and quality of intensivist coverage. Far to be a replacement of the existing model Telemedicine might be a complimentary tool. In fact, to centralize medical data into servers has many additional advantages that could even improve the way in which critical care physicians take care of their patients under the traditional system. PMID- 19811856 TI - [Recommendations for implementing the UNE-EN-ISO 15189 quality standard in the clinical microbiology laboratory: bacteriology and serology]. AB - The UNE-EN-ISO 15189:2007 standard specifies the management and technical requirements that clinical microbiology laboratories must meet to achieve optimal quality when performing microbiological analyses. With implementation of this standard, a laboratory can receive the accreditation and formal recognition of an authorized body, certifying that it is apt for performing an assay or group of assays. In Spain, laboratories that apply these standards can be accredited by the Entidad Nacional de Acreditacion (ENAC, Spanish accreditation body). The purpose of this review is to familiarize clinical microbiology laboratory specialists with the UNE-EN-ISO 15189:2007 standard through a practical approach focussed on bacteriology and serology studies. We briefly define the scope and specify the requisites required for managing the quality of the procedures and processes involved in performing tests on human specimens, for document control, and for management of instruments and equipment, personnel, information systems, supply systems, and external services. Lastly, evaluation approaches are indicated to achieve continuing improvement of the processes carried out and the services the laboratory provides. PMID- 19811857 TI - [Family outbreak of Q fever]. PMID- 19811858 TI - [Ochrobactrum anthropi bacteremia in a child with cystic fibrosis]. PMID- 19811859 TI - A new series of amodiaquine analogues modified in the basic side chain with in vitro antileishmanial and antiplasmodial activity. AB - The synthesis and the study of new amodiaquine derivatives bearing modified lateral basic chains as new agents with both antimalarial and antileishmanial activities are reported. The compounds were tested in vitro against Leishmania donovani MHOM/ET/67/HU3 and 2 strains of Plasmodium falciparum, 3D7 and K1. All the compounds show complex ionisation profiles. At physiological pH the ionised form(s) are in equilibrium with the uncharged form, while at acid pH all the products exist largely as protonated forms. The antiprotozoal profile indicates that all derivatives are endowed with both antimalarial and antileishmanial activity. Interestingly amodiaquine, together with some synthesised derivatives (11, 12, 15, 27, 34), displayed antileishmanial activity in the low micromolar range, although these compounds were also cytotoxic and have a narrow therapeutic window, most of the synthesised compounds proved to be potent antimalarials, a few of them showing a good activity against the chloroquine resistant K1 strain. PMID- 19811860 TI - Masters at work: a narrative inquiry into the experiences of mental health nurses qualifying with an undergraduate Masters degree. AB - The University of Nottingham provides a unique course in the UK that enables undergraduate students to obtain a Masters degree and registration with the Nursing and Midwifery Council (NMC). The curriculum equips graduates with the skills to practice nursing with an analytical attitude and adopt both professional and humanistic values. This research aims to explore significant experiences of graduates, from the undergraduate Masters course, relating to their education and nursing practice in mental health care. A narrative approach to data collection was employed using unstructured individual interviews. Participants worked through a process of contemplation. Their commitment to working with people in a relational manner and studying at graduate level were of high importance. The process continued with assimilation to a philosophy which was intrinsic to the course, including developing therapeutic relationships, self awareness and critical thinking. Participants encountered conflict relating to a perceived dissonance between this philosophy and nursing practice. As a consequence, participants questioned mental health nursing and their abilities as nurses. Resolution occurred when participants were able to work within the constraints of the system whilst effectively realising their philosophy. The findings demonstrate the importance of supportive networks to maintain values and criticality. PMID- 19811861 TI - Machine learning for radioxenon event classification for the Comprehensive Nuclear-Test-Ban Treaty. AB - A method of weapon detection for the Comprehensive nuclear-Test-Ban-Treaty (CTBT) consists of monitoring the amount of radioxenon in the atmosphere by measuring and sampling the activity concentration of (131m)Xe, (133)Xe, (133m)Xe, and (135)Xe by radionuclide monitoring. Several explosion samples were simulated based on real data since the measured data of this type is quite rare. These data sets consisted of different circumstances of a nuclear explosion, and are used as training data sets to establish an effective classification model employing state of-the-art technologies in machine learning. A study was conducted involving classic induction algorithms in machine learning including Naive Bayes, Neural Networks, Decision Trees, k-Nearest Neighbors, and Support Vector Machines, that revealed that they can successfully be used in this practical application. In particular, our studies show that many induction algorithms in machine learning outperform a simple linear discriminator when a signal is found in a high radioxenon background environment. PMID- 19811862 TI - Relation between forest vegetation, atmospheric deposition and site conditions at regional and European scales. AB - Several monitoring programs have been set up to assess effects of atmospheric deposition on forest ecosystems. The aim of the present study was to evaluate effects on the understory vegetation, based on the first round of a regional (the Netherlands) and a European forest monitoring program. A multivariate statistical analysis showed surprisingly similar results for both data sets; the vegetation appeared to be largely determined by the 'traditional' factors soil, climate, and tree species, but there was a small but statistically significant effect of atmospheric deposition. The effects of deposition include a slight shift towards nitrophytic species at high N deposition in the European network, and towards acidophytic species at high S-deposition in the Dutch network. The relatively small effect of atmospheric deposition is understandable in view of the very large natural variation in environmental conditions. Time series of both vegetation and environment are needed to assess deposition effects in detail. PMID- 19811863 TI - Exposure to perfluorooctane sulfonic acid (PFOS) adversely affects the life-cycle of the damselfly Enallagma cyathigerum. AB - We evaluated whether life-time exposure to PFOS affects egg development, hatching, larval development, survival, metamorphosis and body mass of Enallagma cyathigerum (Insecta: Odonata). Eggs and larvae were exposed to five concentrations ranging from 0 to 10000 microg/L. Our results show reduced egg hatching success, slower larval development, greater larval mortality, and decreased metamorphosis success with increasing PFOS concentration. PFOS had no effect on egg developmental time and hatching or on mass of adults. Eggs were the least sensitive stage (NOEC=10000 microg/L). Larval NOEC values were 1000 times smaller (10 microg/L). Successful metamorphosis was the most sensitive response trait studied (NOEC<10 microg/L). The NOEC value suggests that E. cyathigerum is amongst the most sensitive freshwater organisms tested. NOEC for metamorphosis is less than 10-times greater than the ordinary reported environmental concentrations in freshwater, but is more than 200-times smaller than the greatest concentrations measured after accidental releases. PMID- 19811864 TI - Immigrant obesity and unhealthy assimilation: alternative estimates of convergence or divergence, 1995-2005. AB - We re-examine the pace of rising obesity among Hispanic immigrants and the effects associated with longer duration in the US, or what is referred to as unhealthy assimilation, the convergence of immigrant health to a less healthy native-born standard. Consistent with previous research, we find that across all race-ethnic groups, immigrants tend to be less obese than native-born persons. Second, obesity is clearly on the rise, with obesity rates increasing for both immigrant and native-born populations between 1995 and 2005. However, our findings are that immigrant obesity rises more slowly than for native-born Hispanics in the same age cohort. The significance is that immigrants do not converge to obesity prevalence of the native-born as commonly assumed and, in fact, the differential is wider in 2005 than it was in 1995. The analysis, which is based on the National Health Interview Survey tracks the obesity rates of different cohort populations observed in repeated cross-sections (1995 and 2005), as both immigrants and the native-born grow older and additionally, as immigrants reside in the U.S. longer. More specifically, for immigrants, our study distinguishes the effects of length of U.S. residence (observed at a single point in time) and increasing duration of residence (observed over time). Of crucial importance, we contrast the changes over time for native and foreign-born residents passing through the same age range from 1995 to 2005. Misconclusions of previous research stem from 1) assuming that any change for immigrants equates to assimilation, without regard to native-born change, and 2) an unbalanced analysis that fails to track in parallel the growing obesity of both immigrant and native born cohorts. PMID- 19811865 TI - Increased survival with hypotensive resuscitation in a rabbit model of uncontrolled hemorrhagic shock in pregnancy. AB - AIM: We sought to compare the effects of conservative hypotensive and aggressive normotensive resuscitation strategies on blood loss, fluid requirements, blood lactate and survival rate in a clinically relevant model of uncontrolled hemorrhagic shock in pregnancy. METHOD: 60 anesthetized New Zealand white rabbits at late gestation underwent uncontrolled hemorrhagic shock by transecting a small artery in the mesometrium, followed by blood withdrawal via the carotid artery, to a mean arterial pressure (MAP) of 40-45mmHg. They were randomly divided into six groups (n=10 per group): sham shock (group SS); shock without resuscitation (group SH); hypotensive resuscitation in the simulated prehospital phase with Ringer's solution to MAP of 50, 60, or 70mmHg, respectively (groups RE50, RE60, RE70); and aggressive resuscitation in the prehospital phase with Ringer's solution to MAP of 80mmHg (group RE80). Finally, in the simulated hospital phase, animals in the resuscitated groups underwent surgical control of bleeding and were fully resuscitated with half of the heparinized shed blood and Ringer's solution to MAP of 80mmHg. RESULTS: Hypotensive resuscitation significantly decreased blood loss and subsequent volume infusion, leading to higher hematocrit, lower lactate concentration, and shorter prothrombin time and activated partial thromboplastin time. Median survival time in group RE60 (4.3+/ 0.6 days) was significantly longer than that in groups RE50 (2.7+/-0.4 days), RE70 (2.3+/-0.3 days), and RE80 (1.7+/-0.3 days) (P<0.05). CONCLUSIONS: We conclude that in this rabbit model of uncontrolled hemorrhage in pregnancy, hypotensive resuscitation to MAP of 60mmHg may be an optimal target MAP before hemorrhage can be controlled by surgical intervention. PMID- 19811866 TI - Optimum location for chest compressions during two-rescuer infant cardiopulmonary resuscitation. AB - OBJECTIVES: The study aims to evaluate the optimal chest compression site in two rescuer infant cardiopulmonary resuscitation (CPR). METHODS: Charts and multidirectional computed tomography images of infants who presented to one of four hospitals from March 2004 to March 2009 were reviewed retrospectively. The length of the sternum (S(total)), the length and width (L, W) of adult thumbs after two thumbs were placed side-by-side were measured. The study included the structures located underneath the lower third of S(total) (S(total/3)), the lower half of S(total) (S(total/2)), the sternum at the inter-nipple line (S(n)), the point of maximal anterior-posterior heart diameter (S(m)), the lower margin of L and the lateral margin of W from S(total/3), S(total/2), S(n) and S(m). RESULTS: Of the 75 infants enrolled, the ratio of the length from the xiphoid process to S(m) from S(total) was 0.24+/-0.19. In the population studied, 43.1% had aortic roots in S(total/2), 44.0% had left ventricular outflow tracts in S(total/3), 46.7% had left ventricular outflow tracts at S(n) and 100.0% had left ventricles at S(m). All the infants had livers in the lower margin of L from S(m) and all of them had hearts in the left lateral margin of half of W from S(m). A total of 42.7% had lungs in the right lateral margin of half of W from S(m). CONCLUSION: The left ventricle was located in the lower quarter of the sternum, lower than S(total/3). However, more studies are needed to validate the efficiency and safety of compressing the lower quarter of the sternum in two-rescuer infant CPR. PMID- 19811867 TI - Therapeutic hypothermia in cardiac arrest: 206 years later! PMID- 19811868 TI - The length ratios between fetal limb bones and nasal bone in the general population. PMID- 19811869 TI - TERT over-expression affects the growth of myocardial tissue derived from mouse embryonic stem cells. AB - For heart failure, new therapeutic strategies based on augmentation of the regenerative resources of the heart muscle are under evaluation. More knowledge about the mechanisms regulating growth of the embryonic or adult heart muscle will help to improve the results. The present over-expression study provides further insight into the role of telomerase reverse transcriptase (TERT) in growth regulation of myocardial tissue derived from embryonic stem (ES) cells. Mouse ES cells (D3) exhibiting ectopic expression of TERT under the regulation of the beta-actin promoter were generated and allowed to differentiate over a period of up to 18 days. In contrast to the controls, the TRAP assay did not reveal any decrease of telomerase activity during differentiation of TERT transgenic ES cells. Following cell dissociation and staining for sarcomeric alpha-actinin, singular myocardial precursors could be identified and analyzed using fluorescence microscopy: compared with the controls, the outgrowths of TERT transgenic ES cells showed a significant enlargement of the cellular fraction formed by cardiomyocyte precursors, while BrdU-(double) staining did not reveal a change of its proliferation rate. In addition, the average physical dimensions of the precursors appeared to be enlarged. The myocardial precursors exhibited three different morphologies: spindle-like or round or tri-/multi-angular. While, compared with the controls, in TERT transgenic ES cell outgrowths the overall number of myocardial cells was enhanced, the formation of spindle-like or round shaped precursors was suppressed. On the molecular level, RT-PCR analysis showed the mRNA-expression level of alpha-MHC, a gene whose expression is specific for pacemaker-like or atrial-like precursors (Kolossov et al., 2005), to be reduced. Furthermore, TERT transgenic outgrowths displayed a reduced beating frequency. It can be concluded that TERT over-expression promotes the differentiation of mouse ES cell-derived cardiomyocytes in a phenotype-specific manner. PMID- 19811870 TI - Multivariate statistical analysis of heavy metals in street dust of Baoji, NW China. AB - The concentrations of Pb, Cu, Zn, Mn, Ni, Co and Cr in street dust samples from Baoji in north-west China were measured by wavelength dispersive X-ray fluorescence spectrometry, while As and Hg in street dust samples were determined by atomic fluorescence spectrometry. Principal component analysis and cluster analysis, coupled with correlation coefficient analysis, were used to analyze the data and to identify possible sources of these heavy metals. The results indicate that street dust in Baoji has elevated heavy metal concentrations, especially Hg, Pb, Zn and Cu, which are 16-77, 7-92, 6-26 and 4-12 times the background levels in Shaanxi soil, respectively. The mean heavy metal concentrations in street dust divided by the corresponding background values of Shaanxi soil decrease in the order of Hg>Pb>Zn>Cu>Cr>As>Ni>Co>Mn>V. Three main sources of these heavy metals were identified. As, V, Pb and Co originated from nature and traffic. Cu, Zn, Hg and Mn, especially the former two, mainly derive from industry sources, as well as traffic. Cr and Ni mainly originate from soil. PMID- 19811871 TI - Determination of thiourea in fruit juice by a kinetic spectrophotometric method. AB - A catalytic kinetic method is described for determination of trace levels of thiourea based on its catalytic effect on the oxidation of Janus green (JG) by potassium iodate in hydrochloric acid media. The reaction was monitored by measuring the decrease in absorbance of the dye at 610 nm after 25 min. The effect of some factors on the reaction speed was investigated. The developed method allowed the determination of thiourea in range of 0.01-12.00 mg L(-1) with good precision, accuracy and the detection limit was 0.008 mg L(-1). Most of foreign species do not interfere with the determination. The method was found to be sensitive, selective and was applied to the determination of thiourea in fruit juices and orange peel. PMID- 19811872 TI - Influence of soil properties on the sorption and retention of cadmium, copper and lead, separately and together, by 20 soil horizons: comparison of linear regression and tree regression analyses. AB - In this paper we compare linear regression with tree regression for analysis of the influence of soil properties on the sorption and retention of added Cd, Cu and Pb by 20 soil horizons typical of cropped soils in Galicia (N.W. Spain); our measure of sorption/retention capacity was K(r), a recently introduced adimensional parameter. Sorption and retention of Cd was depressed by the presence of Cu and Pb. The soil parameters that were most associated, overall, with differences in Cd, Cu and Pb sorption and retention were cation exchange capacity (CEC), pH and Mn oxides' content. Tree regression, which can take into account variation on both global and local scales, afforded better-fitting models than linear regression, which only reflects global tendencies; but for coherent interpretability of tree regression results it is just as important to avoid overfitting as in the case of linear regression. PMID- 19811873 TI - Numerical study of the effect of water addition on gas explosion. AB - Through amending the SENKIN code of CHEMKIN III chemical kinetics package, a computational model of gas explosion in a constant volume bomb was built, and the detailed reaction mechanism (GRI-Mech 3.0) was adopted. The mole fraction profiles of reactants, some selected free radicals and catastrophic gases in the process of gas explosion were analyzed by this model. Furthermore, through the sensitivity analysis of the reaction mechanism of gas explosion, the dominant reactions that affect gas explosion and the formation of catastrophic gases were found out. At the same time, the inhibition mechanisms of water on gas explosion and the formation of catastrophic gases were analyzed. The results show that the induced explosion time is prolonged, and the mole fractions of reactant species such as CH(4), O(2) and catastrophic gases such as CO, CO(2) and NO are decreased as water is added to the mixed gas. With the water fraction in the mixed gas increasing, the sensitivities of the dominant reactions contributing to CH(4), CO(2) are decreased and the sensitivity coefficients of CH(4), CO and NO mole fractions are also decreased. The inhibition of gas explosion with water addition can be ascribed to the significant decrease of H, O and OH in the process of gas explosion due to the water presence. PMID- 19811874 TI - Sister chromatid exchanges and chromosomal aberrations in Chinese hamster ovary (CHO-K1) cells treated with the insecticide pirimicarb. AB - Pirimicarb and its formulation Aficida (50% pirimicarb) effects were studied on CHO-K1 cells employing sister chromatid exchange (SCE), chromosomal aberrations (CA), cell-cycle progression and mitotic index analyses. Continuous treatments were performed within 10-300 microg/ml concentration-range. Pirimicarb, but not Aficida, induced a concentration-dependent increase of abnormal cells. Pirimicarb induced a greater frequency of chromatid/isochromatid breaks than Aficida did. Regression analyses showed a concentration-dependent increase in the frequency of chromatid-type breaks for both compounds whereas only the frequency of isochromatid-type breaks did in those pirimicarb-treated cultures. SCEs in pirimicarb- or Aficida-treated cultures were significantly higher than control values with concentrations of 100-200 microg/ml. Both test compounds induced equivalent frequency of SCEs. A delay in cell-cycle kinetics was observed for pirimicarb and Aficida within 100-300 and 200-300 microg/ml concentration-range, respectively. An inhibition of MI was observed for both chemicals regardless of tested concentrations. Finally, the CAs appears to be a higher sensitive bioassay to detect DNA damage at lower concentrations of pirimicarb than SCEs does. The results demonstrated that pirimicarb and Aficida exert geno-cytotoxicity, at least in CHO-K1 cells. PMID- 19811875 TI - A comparative study on stabilization of available As in highly contaminated hazardous solid waste. AB - The stabilization of available As was conducted by chemical fixation after Fenton process in a solid waste residual (SWR) from organic arsenic industry. Single as well as combined fixation treatments by using ferric sulfate (FS), magnesium chloride (MC) and calcium hydroxide (CH) were carried out to assess and to evaluate the fixation effect through toxicity characteristic leaching procedure (TCLP), synthetic precipitation leaching procedure (SPLP) and sequential extraction procedure (SEP). The effect of aging treatment on the fixation of available As in SWR was also investigated. Experimental result showed that the optimal molar ratios for Fe:As, Mg:As and Ca:As were 2:1, 3:1 and 2:1, respectively, and the combination fixation FS+MC+CH was found to be the optimal fixation treatment. With respect to the leaching behavior and the speciation migration of As in SWR after stabilization, TCLP, SPLP and SEP represent a pertinent and inseparable system for the fixation effect evaluation. The fixation treatment of available As in SWR could be evaluated directly after 3 days and the aging treatment is not needed though it can further enhance the fixation effect. PMID- 19811876 TI - The natural history of Anaplasma marginale. AB - The intracellular pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae), described by Sir Arnold Theiler in 1910, is endemic worldwide in tropical and subtropical areas. Infection of cattle with A. marginale causes bovine anaplasmosis, a mild to severe hemolytic disease that results in considerable economic loss to both dairy and beef industries. Transmission of A. marginale to cattle occurs biologically by ticks and mechanically by biting flies and by blood contaminated fomites. Both male ticks and cattle hosts become persistently infected with A. marginale and serve as reservoirs of infection. While erythrocytes are the major site of infection in cattle, A. marginale undergoes a complex developmental cycle in ticks that begins by infection of gut cells, and transmission to susceptible hosts occurs from salivary glands during feeding. Major surface proteins (MSPs) play a crucial role in the interaction of A. marginale with host cells, and include adhesion proteins and MSPs from multigene families that undergo antigenic change and selection in cattle, thus contributing to maintenance of persistent infections. Many geographic strains of A. marginale have been identified worldwide, which vary in genotype, antigenic composition, morphology and infectivity for ticks. Isolates of A. marginale may be maintained by independent transmission events and a mechanism of infection/exclusion in cattle and ticks. The increasing numbers of A. marginale genotypes identified in some geographic regions most likely resulted from intensive cattle movement. However, concurrent A. marginale strain infections in cattle was reported, but these strains were more distantly related. Phylogenetic studies of selected geographic isolates of A. marginale, using msp4 and msp1alpha, provided information about the biogeography and evolution of A. marginale, and msp1alpha genotypes appear to have evolved under positive selection pressure. Live and killed vaccines have been used for control of anaplasmosis and both types of vaccines have advantages and disadvantages. Vaccines have effectively prevented clinical anaplasmosis in cattle but have failed to block A. marginale infection. Vaccines are needed that can prevent clinical disease and, simultaneously, prevent infection in cattle and ticks, thus eliminating these hosts as reservoirs of infection. Advances in genomics, proteomics, immunology and biochemical and molecular technologies during the last decade have been applied to research on A. marginale and related organisms, and the recent development of a cell culture system for A. marginale has provided a format for studying the pathogen/tick interface. Recent advancements and new research methodologies should provide additional opportunities for development of new strategies for control and prevention of bovine anaplasmosis. PMID- 19811877 TI - Immune response of bovines stimulated by synthetic vaccine SBm7462 against Rhipicephalus (Boophilus) microplus. AB - Ten-month-old calves Bos taurus taurus were immunized with three doses of SBm7462 with saponin as an adjuvant at 30-day intervals and were evaluated for IgG isotypes, phenotype circulating lymphocytes and changes in the lymph nodes (LN). SBm7462 stimulated the production of predominantly IgG1-isotype IgG antibodies. The lymph nodes exhibited activation at the seventh day after the first immunization, with areas of paracortical and interfollicular hyperplasia and the early formation of germinal centers (GC). Fifteen days after the first immunization, the GC exhibited compartmentalization of cellular populations, a light zone (LZ), a dark zone (DZ) and a mantle. At the same time, hyperplasia of the medullary cords was observed with cells associating with DC cells. Seven days after the first immunization, apoptosis in the DZ and in the paracortical region became evident. By day 15, there was an increase in the medullary cords, which became more numerous at days 35 and 42. PAP-positive cells were found in the paracortical region, medullary cords and GC 7 days after the first immunization. At day 35, there were further strongly PAP-positive cells in the medullary cords. By comparison, none of these changes were observed in the lymph nodes of control groups at any of the days analyzed. The number of CD21(+) lymphocytes increased in the immunized groups after the first inoculation, with a maximum number observed at 15 and 10 days after the first and third immunizations, respectively. Compared to pre-immunization counts, the percentage of WC1(+) gammadelta T lymphocytes displayed more variation, increasing 5 days after the second immunization but decreasing over the following days. According to the results, the synthetic anti Rhipicephalus microplus vaccine elicits a complete immune response being T-dependant. PMID- 19811878 TI - The natural history of Anaplasma phagocytophilum. AB - Anaplasma phagocytophilum is the recently designated name replacing three species of granulocytic bacteria, Ehrlichia phagocytophila, Ehrlichia equi and the agent of human granulocytic ehrlichiosis, after the recent reorganization of the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales. Tick borne fever (TBF), which is caused by the prototype of A. phagocytophilum, was first described in 1932 in Scotland. A similar disease caused by a related granulocytic agent was first described in horses in the USA in 1969; this was followed by the description of two distinct granulocytic agents causing similar diseases in dogs in the USA in 1971 and 1982. Until the discovery of human granulocytic anaplasmosis (HGA) in the USA in 1994, these organisms were thought to be distinct species of bacteria infecting specific domestic animals and free living reservoirs. It is now widely accepted that the agents affecting different animal hosts are variants of the same Gram-negative obligatory intracellular bacterium, which is transmitted by hard ticks belonging to the Ixodes persulcatus complex. One of its fascinating features is that it infects and actively grows in neutrophils by employing an array of mechanisms to subvert their bactericidal activity. It is also able to survive within an apparently immune host by employing a complex mechanism of antigenic variation. Ruminants with TBF and humans with HGA develop severe febrile reaction, bacteraemia and leukopenia due to neutropenia, lymphocytopenia and thrombocytopenia within a week of exposure to a tick bite. Because of the severe haematological disorders lasting for several days and other adverse effects on the host's immune functions, infected animals and humans are more susceptible to other infections. PMID- 19811879 TI - Accelerated postmenopausal cognitive decline is restricted to women with normal BMI: longitudinal evidence from the Betula project. AB - In order to determine whether cognitive performance is influenced by the menopausal transition, we tested cognitive performance at three time points, sampled women in earlier as well as later stages of the menopausal transition (40 65 years of age), and assessed the moderating influence of body mass index (BMI) on rate of change. Multilevel analyses were used to model change in cognitive performance as a function of number of years post menopause over and above chronological age. We investigated change in the menopausal transition for 10 cognitive outcomes in 193 women who were postmenopausal during the last test wave. The model, controlling for age and education, showed that postmenopausal women within the normal range of BMI (BMI 18.5-25) displayed more rapid decline than women with BMI above 25 for measures of visuospatial ability and episodic memory. In addition, there was an accelerated rate of change post menopause for all women on verbal fluency. The results support the notion that the diminished postmenopausal production of endogenous estrogen may have a slight negative influence on cognitive abilities, but mainly for women within a normal BMI range. PMID- 19811881 TI - Managing hereditary ovarian cancer. AB - In this review we present an overview of recent developments in the management of hereditary ovarian cancer. Until recently, intensive screening of the ovaries was recommended to mutation carriers and their first-degree female relatives. However, since screening is not effective in detecting early-stage ovarian cancer, women are counselled for a prophylactic bilateral salpingo-oophorectomy (pBSO) shortly after child-bearing age (>35 years). Many mutation carriers already choose to undergo pBSO to reduce their cancer risks; however, the age of prophylactic surgery may interfere with reproductive and other important (psychosexual) issues in life. Due to the protective effect of oral contraceptives regarding ovarian cancer, we advise women at increased risk of ovarian cancer to use oral contraceptive pills for 3-5 years early in life (<25 years of age), when the absolute incidence of breast cancer is extremely low. A transient increased relative risk of breast cancer due to oral contraceptives at this age will result in a negligible increased absolute number of breast cancers, while the risk reduction of ovarian cancer remains for life. Research should aim at finding new molecular markers and screening strategies for detecting early stage ovarian cancer in women with a hereditary ovarian cancer trait. PMID- 19811880 TI - Clinical factors associated with seroma volume reduction in breast-Conserving Therapy for early-stage breast cancer: a multi-institutional analysis. AB - PURPOSE: To correlate clinical factors with seroma volume and reduction; and to determine whether cone-beam CT (CBCT) could be used clinically to monitor seroma reduction. PATIENTS AND METHODS: This investigation included 102 women from five institutions with stage T1-2 breast cancer treated with breast-conserving therapy. Two CT scans were acquired: the planning CT (CT1) and a second CT (CT2) during radiotherapy (RT). Seroma was contoured on all scans, and correlations between seroma characteristics and clinical factors were investigated by univariate and multivariate analyses. In a substudy, 10 of the 102 patients received multiple CBCT scans during RT. Seroma were contoured by two observers in the substudy. Fifteen time points at which CT and CBCT were performed within 2 days were identified. The levels of correlation in seroma contours between CBCT and CT and between the two observers were examined. RESULTS: The mean relative seroma reduction from CT1 to CT2 was 54% (p < 0.001). A significant inverse relationship was found between relative seroma reduction per week and number of RT fractions given by univariate and multivariate analyses (p = 0.01, 0.03). The mean difference in contoured seroma volumes between CT and CBCT was 12% (3.3 cm(3)). When assessing the relative difference in seroma contours between Observer 1 and Observer 2, an interobserver difference of 12% was demonstrated. Neither discrepancy was clinically significant. CONCLUSIONS: Radiotherapy seems to hinder seroma reduction. Volume discrepancies between CBCT and CT were minor, with low interobserver variation, indicating that CBCT might be useful in monitoring seroma reduction. PMID- 19811882 TI - The frequency of intracranial arterial fenestrations: a study with 64-detector CT angiography. AB - Fenestration is a vascular variation that begins with a common origin, then splits into two parallel luminal channels and rejoins distally. Potential association between anomalies of cerebral circulation and increased occurrence of aneurysm makes intracranial arterial fenestrations important. The planning of intracranial arterial interventions may be complicated if a fenestration occurs proximal to the site of intended treatment. This study is planned to determine the frequency of fenestrations on CT angiography and to search whether there is relationship between aneurysms and fenestrations. CT angiographies of 395 consecutive patients, performed by 64-detector CT, were retrospectively reviewed for aneurysms and fenestrations. Overall fenestration frequency, fenestration frequency in patients with and without aneurysm, and aneurysm frequency in patients with and without fenestration were searched. Demographic characteristics of patients were also compared. Overall fenestration frequency was 12.9%. Vertebrobasilar system (5.56%) and anterior communicating region (5.32%) were the two most frequent sites of fenestration. The rate of fenestrations was not significantly different between patients who had and did not have aneurysms. Mean age was significantly higher, and females were predominant in patients with aneurysms. However our results did not show significant difference in age and sex of patients with fenestrations. The frequency of fenestrations in this study is higher than in previously published radiological studies, suggesting that fenestrations are relatively common. There is no significant relationship between the frequency of aneurysms and fenestrations. PMID- 19811884 TI - Histology after lumpectomy in women with epithelial atypia on stereotactic vacuum assisted breast biopsy. AB - BACKGROUND: Large-core needle biopsy of the breast (LCNB) and vacuum-assisted breast biopsy (VABB) are widely used as alternatives to open surgical biopsy (OSB) for initial diagnosis of mammographic abnormalities. Between 18% and 80% of cases in which such specimens show atypical lobular hyperplasia (ALH) or atypical ductal hyperplasia (ADH) are found to be malignant at surgery. DESIGN: From 1999 to 2005, 68 women with mammographic abnormalities were sampled by stereotactic VABB and presented atypical epithelial hyperplasia. Immunohistochemical staining with anti-cytokeratin 5/6 and anti-E-cadherin antibodies was performed. All women underwent a lumpectomy. Clinical, radiological or histological factors predictive of the risk of finding malignancy at surgery were sought. RESULTS: VABB initially showed 28 cases of ADH, 32 cases of ALH, one case of flat epithelial atypia, five cases of mixed atypia, and two cases of Lobular Carcinoma In Situ (LCIS). After slide review with immunohistochemical staining, two cases of ADH were reclassified as simple hyperplasia and two cases of ALH were reclassified as mixed atypia. Seven lesions (10.3%) that appeared to be benign on VABB were found to be malignant on OSB (Ductal Carcinoma In Situ (DCIS) in six cases and invasive ductal carcinoma in one case). ADH was the only predictive factor of malignancy on OSB (p=0.04 versus ALH). CONCLUSION: ADH diagnosed by vacuum-assisted breast biopsy frequently corresponds to cancer on open surgical biopsy. Surgical excision of all breast lesions containing atypical hyperplasia on percutaneous biopsy can be recommended. PMID- 19811883 TI - Modeling patient-centered communication: oncologist relational communication and patient communication involvement in breast cancer adjuvant therapy decision making. AB - OBJECTIVE: Relational communication refers to those messages communicators naturally express that carry meaning about the type and quality of relationship they share. It is expected that patients of oncologists who express positive relational communication will be more communicatively involved in their office visits, and regret their decision for adjuvant therapy following surgery less. METHODS: One hundred eighty (180) audio-recorded discussions between oncologists (n=40) and early stage (I-III) breast cancer patients were coded with the Siminoff Communication Content and Affect Program (SCCAP). The data were used to test the relationships between patient demographics, oncologist relational communication, patient communication involvement and self-reported patient decision regret. RESULTS: After controlling for clinician clusters, oncologists' verbal (i.e., confirming messages) and nonverbal (i.e., direct and inclusive speech) relational communication is indirectly associated with lower patient decision regret via the mediating effect of greater patient communication involvement. CONCLUSION: Clinician relational communication provides an influential affective climate for decision-making that appears to have important effects on patients' decision confidence. PRACTICE IMPLICATIONS: Clinicians should recognize the potential of their own relational messages to facilitate patients' communication involvement in decision-making during cancer care. PMID- 19811885 TI - Atorvastatin inhibits inflammatory angiogenesis in mice through down regulation of VEGF, TNF-alpha and TGF-beta1. AB - While compelling evidence indicates beneficial effects of statins on inflammatory processes, besides their cholesterol-lowering activities, the actions on angiogenesis are less clear-cut. Our aim was to investigate the effects of atorvastatin on key components of inflammatory angiogenesis in the murine sponge model. Polyester-polyurethane sponges, used as a framework for fibrovascular tissue growth, were implanted in Swiss mice. Atorvastatin (0.6, 3 mg/kg/day) was given orally for 8 days in drinking water. The implants collected at day 9 postimplantation were processed for the assessment of hemoglobin, myeloperoxidase (MPO), N-acetylglucosaminidase (NAG) and collagen. Relevant inflammatory, angiogenic and fibrogenic cytokines were also determined. Atorvastatin treatment resulted in significant decrease in sponge vascularization (Hb content) and in VEGF levels at both doses. Neutrophil influx (MPO activity) was not affected by the compound whereas macrophage recruitment (NAG activity) was inhibited, suggesting a degree of selectivity by atorvastatin for this cell population. The level of CCL2 (MCP1-JE) was decreased only with 0.6 mg/kg. Atorvastatin was also able to reduce collagen deposition and the levels of transforming growth factor (TGF-beta1) intraimplant, dose-dependently. The inhibitory function of atorvastatin on multiple parameters of main components of inflammatory angiogenesis revealed in this study is clearly associated with the modulatory effects of HMG-CoA reductase on VEGF, TNF-alpha and TGF-beta1 production. PMID- 19811886 TI - Symptom experience in HIV-infected adults: a function of demographic and clinical characteristics. AB - Personal characteristics that interact with both HIV diagnosis and its medical management can influence symptom experience. Little is known about how symptoms in populations with chronic illness vary by age, sex, or socioeconomic factors. As part of an ongoing prospective longitudinal study, this report describes symptoms experienced by 317 men and women living with HIV/AIDS. Participants were recruited at HIV clinics and community sites in the San Francisco Bay Area. Measures included the most recent CD4 cell count and viral load from the medical record, demographic and treatment variables, and the 32-item Memorial Symptom Assessment Scale to estimate prevalence, severity, and distress of each symptom and global symptom burden. The median number of symptoms was nine, and symptoms experienced by more than half the sample population included lack of energy (65%), drowsiness (57%), difficulty sleeping (56%), and pain (55%). Global symptom burden was unrelated to age or CD4 cell count. Those with an AIDS diagnosis had significantly higher symptom burden scores, as did those currently receiving antiretroviral therapy. African Americans reported fewer symptoms than Caucasians or Mixed/Other race, and women reported more symptom burden after controlling for AIDS diagnosis and race. Because high symptom burden is more likely to precipitate self-care strategies that may potentially be ineffective, strategies for symptom management would be better guided by tailored interventions from health care providers. PMID- 19811887 TI - Goals of care and end-of-life decision making for hospitalized patients at a canadian tertiary care cancer center. AB - Limited information is available regarding the quality of end-of-life care at cancer centers. We sought to characterize the end-of-life decision-making process for advanced cancer patients admitted to our tertiary cancer center, and to examine the association between goals of care and practice patterns. Information on patient characteristics, investigations, cancer treatments, and goals of care was collected retrospectively for consecutive patients who died at the inpatient unit of the Vancouver Cancer Center between January 1, 2005 and December 31, 2006. One hundred eighteen advanced cancer patients had a median admission duration of 10 days (range 1-64 days). A median of two tests per day was performed, with a decreasing trend over time (P<0.001). Forty percent received cancer treatments during hospitalization, with 75% terminated prematurely. Do-not resuscitate orders, supportive care plans, and diagnosis of dying were documented for 96%, 86%, and 76% of the patients, respectively. Early establishment of supportive care plan and diagnosis of dying were associated with timely discontinuation of cancer treatments (Spearman coefficients 0.47 and 0.60, respectively). Multivariate analysis revealed that timely diagnosis of dying was associated with early establishment of code status (P=0.042), supportive care plans (P<0.001), and discontinuation of cancer therapy (P=0.005). Cancer patients who died at our oncology center were investigated and treated intensively during their short hospitalization. Early establishment of goals of care may be associated with changes in practice consistent with improved quality of care. PMID- 19811888 TI - Poststroke fatigue--a review. AB - Although fatigue is a common complaint after stroke, relatively little is known about how poststroke fatigue is experienced and what its related factors are. An in-depth understanding is necessary to develop effective and patient-centered poststroke rehabilitation programs. This review was undertaken to provide a comprehensive synthesis of knowledge from the literature concerning the description, definition, and measurement of fatigue and its relationship to sociodemographic and clinical factors. A search in PubMed, CINAHL, EMBASE, and PsychInfo was performed using "stroke" or "cerebrovascular accident" as medical subject headings in combination with "fatigue" as a key word. Descriptions of fatigue revealed multiple dimensions of the phenomenon. Although no specific theoretical definition of fatigue as a poststroke condition was found, a case definition has recently been published to be used as a tool to determine the presence of fatigue in poststroke patients. Poststroke fatigue is most frequently measured by using the general fatigue scales such as the Fatigue Severity Scale and a Fatigue Visual Analogue Scale, as there is no scale developed to measure poststroke fatigue specifically. Age, sex, living conditions, and personality were associated with poststroke fatigue, albeit with some conflicting findings. Conflicting results also were found in the relationships between fatigue and stroke-related characteristics such as stroke location/type, the number of strokes, and neurological deficits. There is an indication that prestroke and poststroke fatigue are related. Possible antecedent components identified are personal factors, biomarkers, stroke characteristics, prestroke fatigue, and comorbidity. As knowledge regarding poststroke fatigue remains limited, there is a need to continue empirical research with various theoretical orientations. PMID- 19811889 TI - Assisting people with developmental disabilities to improve pointing efficiency with a Dual Cursor Automatic Pointing Assistive Program. AB - This study evaluated whether two persons with developmental disabilities would be able to improve their pointing performance through a Dual Cursor Automatic Pointing Assistive Program (DCAPAP) with a newly developed mouse driver (i.e., a new mouse driver replaces standard mouse driver, and is able to intercept/detect mouse movement action). First, baseline sessions started with both participants. Then the first participant had his intervention session. When his performance was consolidated, new baseline and intervention occurred with the second participant. Finally, both participants were exposed to maintenance phase, in which their pointing performance improved significantly. Data showed that both participants improved their pointing efficiency through the use of DCAPAP and retained their successful results through maintenance phase. Implications of the findings are discussed. PMID- 19811890 TI - Differential effects of voice therapies on neurovegetative symptoms and complaints. AB - In previous studies, female patients in all age categories with a nonorganic dysphonia were found to report significantly more autonomic symptoms and complaints than healthy controls. After voice therapy, there was a highly significant reduction in the amount of autonomic symptoms and complaints (related or not related to voice). The present prospective study with a matched control group is designed to test the hypothesis that a specific kind of therapy is more efficient than the usual approaches in reducing these neurovegetative symptoms and complaints. Two matched groups of 34 patients diagnosed with nonorganic dysphonia and referred for voice therapy answered a questionnaire of 46 questions with 3 subsets and a consistency control. They received either "coordination therapy" (CTh)--a holistic approach addressing functional, personal and emotional aspects--or a conventional voice therapy (approximately 15 sessions). All patients again filled in a similar questionnaire after approximately 6 months. After therapy, there is in general a highly significant reduction in the amount of autonomic symptoms and complaints (related or not related to voice), to such an extent that patients report on average no more general neurovegetative symptoms and complaints than those of healthy controls. Symptoms and complaints of other nature (validity control) are not influenced. When compared with patients receiving conventional therapy, those who received CTh demonstrate a significantly higher reduction for the subset "neurovegetative symptoms/complaints related to voice and speech." PMID- 19811891 TI - Voice in chronic renal failure. AB - Chronic renal failure affects various body systems, one of which is the respiratory system. Because respiration is the prime source for speech, vocal dysfunctions are expected to be present in patients with chronic renal failure. The present study attempts to shed light on the changes in acoustic and aerodynamic characteristics of voice, if any, in patients with chronic renal failure. Phonation of sustained vowel /a/ was subjected to acoustic analysis using VAGHMI software (Voice and Speech Systems, Bangalore, Karnataka, India). Sustained phonation of /a/, /s/, and /z/ was recorded for the purpose of aerodynamic analysis. Independent t test was used to find the significant difference between the two groups. Chronic renal failure subjects showed significant deviation in frequency, perturbation, and aerodynamic measures when compared with normal subjects. These results are discussed with respect to the underlying pathophysiology. PMID- 19811892 TI - Biased ART: a neural architecture that shifts attention toward previously disregarded features following an incorrect prediction. AB - Memories in Adaptive Resonance Theory (ART) networks are based on matched patterns that focus attention on those portions of bottom-up inputs that match active top-down expectations. While this learning strategy has proved successful for both brain models and applications, computational examples show that attention to early critical features may later distort memory representations during online fast learning. For supervised learning, biased ARTMAP (bARTMAP) solves the problem of over-emphasis on early critical features by directing attention away from previously attended features after the system makes a predictive error. Small-scale, hand-computed analog and binary examples illustrate key model dynamics. Two-dimensional simulation examples demonstrate the evolution of bARTMAP memories as they are learned online. Benchmark simulations show that featural biasing also improves performance on large-scale examples. One example, which predicts movie genres and is based, in part, on the Netflix Prize database, was developed for this project. Both first principles and consistent performance improvements on all simulation studies suggest that featural biasing should be incorporated by default in all ARTMAP systems. Benchmark datasets and bARTMAP code are available from the CNS Technology Lab Website: http://techlab.bu.edu/bART/. PMID- 19811895 TI - Effect of biomedical organic compounds on the setting reaction of calcium phosphates. AB - In the present study, the effect of biomedical organic compounds (starch, sodium alginate, chitosan and gelatin) on the hydration of calcium phosphates was studied using X-ray diffraction, infrared spectroscopy, scanning electron microscopy and XPS analysis. Amorphous calcium phosphate (ACP) was prepared by a mechanochemical route and mixed with biomedical organic compounds. A solidification reaction occurred between ACP and dicalcium phosphate dihydrate (DCPD); the hydration product was poorly crystallized hydroxyapatite (HA). During the setting reaction of ACP and DCPD, the presence of biomedical organic compounds had an effect on the hydration product: the bonding energies of the primary elements (Ca, P) in the hydration product (HA) were changed; also different hydration morphologies, self-setting properties, rheological properties and mechanical strength of the cement were obtained. This work will allow advances in the synthesis of bionic composite calcium phosphate cement (CPC). PMID- 19811894 TI - Effect of high-fat feeding on expression of genes controlling availability of dopamine in mouse hypothalamus. AB - OBJECTIVE: Hypothalamic centers integrate external signals of nutrient availability and energy status and initiate responses to maintain homeostasis. Quantifying changes in hypothalamic gene expression in the presence of nutrient excess may identify novel responsive elements. METHODS: Affymetrix Mouse Genome 430 2.0 oligonucleotide microarrays containing 45 102 probe sets were used to interrogate differential expression of genes in dietary-induced obesity model C57BL6 inbred mice fed a high-fat (35% fat; n=8) or standard (4% fat; n=6) diet from 3 to 15 wk of age. Ontologies of regulated genes were examined and expression of selected genes was validated by quantitative real-time polymerase chain reaction. RESULTS: One thousand two hundred twelve unique gene transcripts showed altered expression on the microarrays. Gene ontology analysis revealed changes in neuropeptide genes responding to leptin, Pomc, Cart, Npy, and Agrp, compatible with a homeostatic response to high-fat intake, although mean weight increased 2.3-fold compared with standard fed mice (P<0.001). Neurotransmitter system ontologies revealed upregulation of five genes controlling availability of dopamine. Changes in Th tyrosine hydroxylase (2.1-fold) and Slc18a2 solute carrier family 18 (vesicular monoamine), member 2 (4.4-fold) controlling synthesis and release, and Slc6a3 solute carrier family 6 (neurotransmitter transporter, dopamine), member 3 (4.8-fold), Snca alpha-synuclein (1.3-fold), and Maoa monoamine oxidase (1.9-fold) limiting availability were confirmed by quantitative real-time polymerase chain reaction. CONCLUSION: Expression of five genes involved in availability of dopamine was increased after a high-fat diet. Failure to reduce dopamine availability sufficiently, to counter the feeding reward effect, could contribute to diet-induced obesity in these mice. PMID- 19811896 TI - How surface composition of high milk proteins powders is influenced by spray drying temperature. AB - High milk proteins powders are common ingredients in many food products. The surface composition of these powders is expected to play an essential role during their storage, handling and/or final application. Therefore, an eventual control of the surface composition by modifying the spray-drying temperature could be very useful in the improvement of powder quality and the development of new applications. For this purpose, the influence of five spray-drying temperatures upon the surface composition of the powders was investigated by X-ray photoelectron spectroscopy. The major milk proteins were studied: native micellar casein and native whey, both more or less enriched in lactose. The results show a surface enrichment in lipids for all the powders and in proteins for many powders. Whatever the drying temperature, lipids and proteins are preferentially located near the surface whereas lactose is found in the core. This surface enrichment is also highly affected by the spray-drying temperature. More lipids, more proteins and less lactose are systematically observed at the surface of powders spray-dried at lower outlet air temperatures. The nature of proteins is also found essential; surface enrichment in lipids being much stronger for whey proteins containing powders than for casein containing powders. Additionally, we found a direct correlation between the lipids surface concentration and the wetting ability for the 25 powders studied. PMID- 19811893 TI - Spontaneous autoimmune myocarditis and cardiomyopathy in HLA-DQ8.NODAbo transgenic mice. AB - Most individuals have viral infections at some point in their life, however, only few develop autoreactivity to cardiac myosin following infection suggesting a genetic predisposition. Population studies have shown that among all the genetic factors linked with autoimmune disease development, MHC class II genes are the most significant genetic factors. Experimental autoimmune myocarditis resembling human Dilated cardiomyopathy can be induced in susceptible mice by infection with coxsackie virus as well as immunization with purified foreign and murine cardiac specific a-myosin. We generated transgenic mice lacking endogenous class II molecules, HLA-DR3.Abo and HLA-DQ8.Abo transgenic mice in NOD and HLA-DQ8.Abo in B10 background, to study the role of MHC in spontaneous autoimmunity. The HLA molecules in these mice are expressed on cell surface and can positively select CD4+ T cells expressing various Vb T cell receptors. NOD.DQ8 female mice spontaneously developed myocarditis and dilated cardiomyopathy. Histopathology of heart revealed mononuclear infiltrate consisting of CD4 and Mac-1+ cells and myocyte necrosis. NOD.DQ8 mice showed cellular and humoral autoreactive response to self cardiac myosin.. Depletion of CD8 and CD4 + cells suggested that CD8 T cells may act as regulatory cells while CD4 cells are required as effector cells. NOD.DR3 and B10.DQ8 mice did not develop any cardiac pathology suggesting DQ8 is required for predisposition to the spontaneous autoreactivity while NOD background influences onset and progression of disease. Thus these mice provide powerful tools to understand the role of HLA class II molecules in predisposition and onset of human diseases and to develop immunotherapy. PMID- 19811897 TI - Blood compatibility evaluation of poly(D,L-lactide-co-beta-malic acid) modified with the GRGDS sequence. AB - Endothelialization is an ideal approach to improve the blood compatibility of synthetic polymers. However, cell detachment is inevitable under shear flow conditions. Therefore, the issue of blood compatibility needs to be addressed for both the bare and the endothelialized polymer. RGD-containing polymer P-GS5 was synthesized by modification of poly(D,L-lactide-co-beta-malic acid) (PLMA) with the peptide GRGDS. The compositions, molecular weights and hydrophilicities of poly(D,L-lactide) (PDLLA), PLMA, and P-GS5 were characterized by nuclear magnetic resonance (NMR), X-ray photoelectron spectroscopy (XPS), gel-permeation chromatography (GPC) and water contact angle measurements, respectively. The blood compatibilities of the bare and the endothelialized polymers were evaluated by clotting time and platelet adhesion tests. The results showed that the coagulation pathways were not influenced before and after cell culture; the bare P-GS5 attracted less platelet adhesion and induced lower pseudopodia extension compared with PDLLA and PLMA, and the platelet adhesion on P-GS5 was almost completely eliminated after cell seeding. The results suggest that P-GS5 could be a potentially useful material in vascular tissue engineering. PMID- 19811898 TI - [Screening and management of glucose metabolism disorders in cystic fibrosis patients. Practices survey in 4 French reference centers]. AB - OBJECTIVE: Glucose metabolism disorders are a new point of interest in cystic fibrosis (CF) management. Cystic fibrosis-related diabetes mellitus (CFRD) increases alteration of pulmonary function as well as patients' morbidity and mortality. In France, CF patients are exclusively followed up in reference centers. We conducted a practices survey on screening and diagnosis of glucose metabolism disorders at 4 French CF centers. The objective of this study was to assess adherence to practice guidelines developed in 2002 at these centers. METHODS: This study was conducted in 2 sessions: 60 medical records were randomly selected in 2005 and in 2007 for patients aged over 10 years followed up at 4 CF centers. A questionnaire survey was completed for each patient with questions on CFRD screening, diagnosis, monitoring and treatment. Our guidelines recommend random blood glucose (RBG) at each standard biological test, annual glycosylated haemoglobin and an oral glucose tolerance test (OGTT) at 10, 15 and 18 years of age, then every 2 or 3 years. RESULTS: An annual RBG was performed in 82% of patients in 2005 and 91.5% in 2007. HbA1c screening was performed annually for 77% of patients in the 1st session and for 90% of patients for the 2nd session (p<0.10). Adherence to OGTT guidelines was better for adults than children: 96% had an OGTT during the 3 years of the first session and 79% during the second session, while fewer than 50% of children had their OGTT at 15 and 18 years of age. Taking the OGTT at 10 years of age could not be assessed because no patients were 10 years old during the study period. Screening for neurological complications of CFRD was assessed in the majority of diabetic patients, while half or less than half had annual fundus oculi or microalbuminuria dosage. DISCUSSION: There was an improvement in screening for CFRD and glucose metabolism disorders between 2005 and 2007, even though practices could still be improved. This shows that clinical guidelines can be implemented and followed. However, screening and management criteria for glucose metabolism disorders must be consensus-based with higher evidence in order to limit the variability of practices and prevent CFRD-related complications. PMID- 19811899 TI - Serum KL-6 in fibrotic NSIP: Correlations with physiologic and radiologic parameters. AB - BACKGROUNDS: Fibrotic nonspecific interstitial pneumonia (f-NSIP) has been recognized as a distinct disease entity. KL-6 has been reported to be a useful serum marker in interstitial lung diseases. However, few previous reports evaluated the value of serum KL-6 exclusively in f-NSIP, as distinct from other subtypes of idiopathic interstitial pneumonia, therefore the associations of serum KL-6 with clinical and radiologic findings in this population remain unclear. METHODS: Serum KL-6 levels were measured in twenty-six consecutive patients with f-NSIP diagnosed by surgical lung biopsy. Pulmonary function testing, bronchoalveolar lavage, subjective measurement of dyspnea using baseline dyspnea index (BDI), and HRCT were performed in parallel. Two radiologists conducted independent visual examinations of the pattern and extent of abnormalities on HRCT. RESULTS: Serum KL-6 levels were elevated above the cut-off level in all patients. In univariate analysis serum KL-6 levels showed negative correlations with BDI (rho=-0.52; p<0.01). Serum KL-6 had positive correlations with the extent of several patterns of opacities (rho=0.56-0.62; p<0.01). Among them, only the extent of traction bronchiectasis in HRCT showed significant association with serum KL-6 in multivariate analysis (beta-coefficient=0.043; p<0.01). CONCLUSIONS: Serum levels of KL-6 were elevated in f-NSIP, and were correlated with the extent of fibrotic abnormalities on HRCT, suggesting a value of serum KL-6 as a marker for fibrosis in f-NSIP. PMID- 19811900 TI - A review of mechanochemistry applications in waste management. AB - Mechanochemistry is defined to describe the chemical and physicochemical transformation of substances during the aggregation caused by the mechanical energy. Mechanochemical technology has several advantages, such as simple process, ecological safety and the possibility of obtaining a product in the metastable state. It potentially has a prospective application in pollution remediation and waste management. Therefore, this paper aims to give an overall review of the mechanochemistry applications in waste management and the related mechanisms. Based on our study, the modification of fly ash and asbestos containing wastes (ACWs) can be achieved by mechanochemical technology. Waste metal oxides can be transformed into easily recyclable sulfide by mechanochemical sulfidization. Besides, the waste plastics and rubbers, which are usually very difficult to be recycled, can also be recycled by mechanochemical technology. PMID- 19811901 TI - Nitrification of ammonium-rich sanitary landfill leachate. AB - The nitrification of ammonium-rich wastewater is considered challenging due to the substrate inhibition particularly in the form of free ammonia (FA) and free nitrous acid (FNA) in ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). The feasibility of the nitrifying activated sludge system to completely nitrify synthetic stabilized landfill leachate with N-NH(4)(+) concentration of 1452mg/L was tested in this study. The process started with 0.4kg N-NH(4)(+)/m(3)/day of nitrogen loading rate (NLR) in a fed-batch mode to avoid any accumulation of the FA and FNA in the system followed by increasing the nitrogen loading rate (NLR) gradually. Complete nitrification was achieved with a very high ammonium removal percentage (approximately 100%). The maximum specific and volumetric nitrification rate obtained were 0.49g N-NH(4)(+)/g VSS/day and 3.0kg N-NH(4)(+)/m(3)/day, respectively which were higher than those reported previously for ammonium-rich removal using activated sludge system. The nitrifying sludge exhibited good settling characteristics of up to 36mL/g VSS and a long SRT of more than 53 days which contributed to the success of the nitrification process. The coexistence and syntrophic association of the AOB and NOB was observed by using Fluorescence in situ hybridization (FISH) technique which supported the results on complete nitrification obtained in the system. These findings would be of prominent importance for further treatment of actual sanitary landfill leachate. PMID- 19811902 TI - Balancing dormant and self-renewing hematopoietic stem cells. AB - The mouse hematopoietic stem cell (HSC) is probably the best-understood somatic stem cell in higher organisms. Recent studies have shown that the highest self renewal potential is most likely contained within an exceedingly small number of deeply dormant bone marrow HSCs. These stem cells are housed in individual niches that preserve their dormancy via signaling molecules such as Thrombopoietin, Angiopoietins, and Stem Cell Factor. In response to injury cues, dormant HSCs are efficiently activated and produce numerous progenitors and mature cells. A series of intracellular regulatory molecules including FoxOs, mTORC1, Fbw7, Egr1, Pbx1, pRb, c-Cbl, Myc, and Bmi1 mediate the processes of dormancy, cycling, self renewal, differentiation, and survival, all of which control the behavior of HSCs. PMID- 19811903 TI - The structural changes of T7 RNA polymerase from transcription initiation to elongation. AB - The structures of T7 RNA polymerase (T7 RNAP) captured in the initiation and elongation phases of transcription, as well as an intermediate stage provide insights into how this RNA polymerase protein can initiate RNA synthesis and synthesize 7-10 nucleotides of RNA while remaining bound to the DNA promoter site. Recently, the structures of T7 RNAP bound to its promoter DNA along with either a seven nucleotide or eight nucleotide transcript show an elongated product site resulting from a 40 degrees or 45 degrees rotation of the promoter and domain that binds it. The different functional properties of the initiation and elongation phases of transcription are illuminated from structures of the initiation and elongation complexes. Structural insights into the translocation of the product transcript of RNAP, its separation of the downstream duplex DNA, and its removal of the transcript from the heteroduplex are provided by the structures of several states of nucleotide incorporation. A conformational change in the 'fingers' domain that results from the binding or dissociation of incoming NTP or PPi appears to be associated with the state of translocation of T7 RNAP. PMID- 19811904 TI - New opportunities for drug outcomes research in cancer patients: the linkage of the Eindhoven Cancer Registry and the PHARMO Record Linkage System. AB - BACKGROUND: Insight into co-morbidity and treatment effects is pivotal to improve quality of care for cancer patients. OBJECTIVES: To determine whether linkage of the Eindhoven Cancer Registry (ECR) and the PHARMO Record Linkage System (RLS) was technically feasible and to assess which patient-centric data would result from this linkage. METHODS: The ECR records data on tumour stage and primary treatment of all newly diagnosed cancer patients in the southeastern Netherlands including co-morbidity at diagnosis, whereas the PHARMO RLS includes data from multiple linked observational databases such as data on drug utilisation (for both in- and out-patients, including chemotherapy), hospitalisations and clinical laboratory measurements. All patients who lived or had been living in the overlapping area served by the ECR and the PHARMO RLS during 1998-2006 were selected for linkage which was performed with probabilistic medical record linkage. RESULTS: The linkage resulted in an ECR-PHARMO cohort of 40,004 cancer patients with a total of 42,767 primary tumours. The cancer patients in the linked ECR-PHARMO cohort were representatives for the cancer patients included in the total ECR during 1998-2006. Cancer patients included in the cohorts had a mean history of 5 years and a mean follow-up ranging from 2 to more than 4 years (dependent on the survival rate of the specific cancer type). CONCLUSIONS: Linkage of ECR and the PHARMO RLS creates the possibility to study patient centric drug utilisation, health resources utilisation and their costs, in addition to the effectiveness and safety of pharmaceuticals in routine daily practice in cancer patients. PMID- 19811905 TI - Prevalence, socio-demographic and clinical predictors of post-diagnostic utilisation of different types of complementary and alternative medicine (CAM) in a nationwide cohort of Danish women treated for primary breast cancer. AB - AIM: This study investigated the prevalence and predictors of use of complementary and alternative medicine (CAM) in a nationwide inception cohort of Danish women treated for early-stage breast cancer as well as differences in user patterns for individual types of CAM. METHODS: Use of CAM since the time of diagnosis was assessed 12-16 weeks post-surgery for the 3343 women (age 18-70) included in the study (response rate: 68%). Socio-demographic and clinical variables were obtained from national longitudinal registries. RESULTS: 40.1% of the women had used one or more types of CAM. Users were younger than non-users. Age adjusted analyses showed that CAM users were characterised by absence of comorbidity, higher educational level, higher personal income, higher social status, being divorced/separated and living in the metropolitan area of Copenhagen. Multivariate analyses revealed that chemotherapy was the only clinical and treatment-related predictor of CAM use, and that CAM users were more likely to be of normal weight and non-smokers. Of CAM users, 33.7% believed that CAM would have a positive influence upon their breast cancer. Different characteristics distinguished users of individual types of CAM. CONCLUSION: The results of this first nationwide study of utilisation of CAM in breast cancer show that CAM users are healthier and more likely to have higher socio-economic status than non-users. Different user patterns for individual types of CAM may be overlooked, when different types of CAM are treated as one homogeneous category. PMID- 19811906 TI - The ratio of maximum percent tumour accumulations of the pretargeting agent and the radiolabelled effector is independent of tumour size. AB - Our previous studies have indicated that the optimal dosage ratio of pretargeting antibody to effector is proportional to their maximum percent tumour accumulations (MPTAs). This study quantitatively describes how both MPTAs and their ratio change with tumour size, to simplify pretargeting optimisation when tumour size varies. The CC49 antibody dosages below saturation of the tumour antigen level were first examined for the LS174T tumour mouse model. Then the MPTAs of the antibody in mice bearing tumours of different sizes were determined, always at antibody dosages below antigen saturation. Historical data from this laboratory were used to collect the MPTAs of the (99m)Tc-cMORF effector for different tumour sizes, always at effector dosages below that required to saturate the MORF in tumour. The MPTAs versus tumour sizes for both the antibody and the effector were fitted non-linearly. The best fit of the antibody MPTA (Y(antibody)) with tumour size (x) in grams was Y(antibody)=19.00 x(-0.65) while that for the effector was Y(effector)=4.51x(-0.66). Thus, even though the MPTAs of both vary with tumour size, the ratio (Y(antibody)/Y(effector)) is a constant at 4.21. In conclusion, the MPTA ratio of the antibody to the effector was found to be constant with tumour size, an observation that will simplify pretargeting optimisation because remeasurement of the optimum dosage ratio for different tumour sizes can be avoided. Theoretical considerations also suggest that this relationship may be universal for alternative antibody/effector pairs and for different target models, but this must be experimentally confirmed. PMID- 19811907 TI - ONCOPOOL - a European database for 16,944 cases of breast cancer. AB - ONCOPOOL is a retrospectively compiled database of primary operable invasive breast cancers treated in the 1990s in 10 European breast cancer Units. Sixteen thousand and nine hundred and forty four cases were entered, with tumours less than 5 cm diameter in women aged 70 or less (mean age 55). DATA: Data were date of birth, mode of diagnosis, pathology (size, lymph node status, grade, type, lympho-vascular invasion and hormone receptor) and therapies and outcome measures: first local, regional or distant recurrences, contralateral primary, date and cause of death. TUMOUR CHARACTERISTICS: Mean diameter 1.8 cm, 66% lymph node negative, 24% 1-3 lymph nodes involved and 10% had 4 or more involved. Grade 1, 29%; Grade 2, 41%; and Grade 3, 30%. Polynomial relationships were established between grade, stage and size. Seventy-five percent were oestrogen receptor (ER) positive. ER closely related to grade. OUTCOMES: Overall Survival was 89% at 5 years from diagnosis, 80% 10 years and 73% 15 years; Breast Cancer-Specific survivals were 91%, 84% and 79%. Survival strongly related to the Nottingham Prognostic Index (NPI). Cases detected at screening had 84% 10-year survival, those presenting symptomatically 76%. ER positive cases treated with adjuvant hormone therapy had a reduction in risk of death of 13% over those not receiving adjuvant therapy (p=0.000). ER negative cases treated with chemotherapy showed a risk reduction of 23% over those not receiving chemotherapy (p=0.000). PMID- 19811908 TI - An assessment of the characteristics of yard trimmings and recirculated yard trimmings used in biowaste composting. AB - The aim of this research was to characterise samples of yard trimmings (YT) and recirculated yard trimmings (RYT) that are used to co-compost organic fraction of municipal solid waste (OFMSW) taken from 18 composting facilities in Catalonia (NE Spain), to determine their contribution in the composting process, and to highlight the advantages and disadvantages of reusing RYT. We found significant differences between the characteristics of the two materials. RYT had higher pH levels, electrical conductivity, and organic and ammonium nitrogen content than YT. Nutrient content also shows an increment from YT to RYT, and in the case of P and K, this variation can be attributed to an exogenous source rather than the relative concentration during the composting process. We also found significant differences in particle size distribution and bulk density. From the results, it can be assumed that RYT could be reused in the composting process if OFMSW, YT, and RYT are used in the correct quantities to balance the mixture but they cannot totally replace the features of YT. PMID- 19811909 TI - Process optimization to convert forage and sweet sorghum bagasse to ethanol based on ammonia fiber expansion (AFEX) pretreatment. AB - With growing demand for bio-based fuels and chemicals, there has been much attention given to the performance of different feedstocks. We have optimized the ammonia fiber expansion (AFEX) pretreatment and fermentation process to convert forage and sweet sorghum bagasse to ethanol. AFEX pretreatment was optimized for forage sorghum and sweet sorghum bagasse. Supplementing xylanase with cellulase during enzymatic hydrolysis increased both glucan and xylan conversion to 90% at 1% glucan loading. High solid loading hydrolyzates from the optimized AFEX conditions were fermented using Saccharomyces cerevisiae 424A (LNH-ST) without any external nutrient supplementation or detoxification. The strain was better able to utilize xylose at pH 6.0 than at pH 4.8, but glycerol production was higher for the former pH than the latter. The maximum final ethanol concentration in the fermentation broth was 30.9 g/L (forage sorghum) and 42.3 g/L (sweet sorghum bagasse). A complete mass balance for the process is given. PMID- 19811910 TI - Multi-stage continuous culture fermentation of glucose-xylose mixtures to fuel ethanol using genetically engineered Saccharomyces cerevisiae 424A. AB - Multi-stage continuous (chemostat) culture fermentation (MCCF) with variable fermentor volumes was carried out to study the utilization of glucose and xylose for ethanol production via mixed sugar fermentation (MSF). Variable fermentor volumes were used to enable enhanced sugar utilization, accounting for differences in glucose and xylose utilization rates. Saccharomyces cerevisiae 424A-LNH-ST was used for fermentation of glucose-xylose mixtures. The dilution rates employed for continuous fermentation were based on earlier batch kinetic studies of ethanol production and sugar utilization. With a feed containing approximately 30 g L(-1) glucose and 15 g L(-1) xylose, cell washout was observed at a dilution rate of 0.8 h(-1). At dilution rates below 0.5 h(-1), complete glucose utilization was observed. Xylose consumption in the first-stage 1 L reactor was only 37% at the lowest dilution rate studied, 0.0 5h(-1). At this same flow rate, xylose consumption rose to 69% after subsequently passing through 3 and 1 L reactors in series, primarily due to the longer residence time in the 3 L reactor (0.0167 h(-1) dilution rate). PMID- 19811911 TI - Synthesis and biological activity of novel oxazolidinones. AB - A number of 5-substituted derivatives of Ranbezolid, a novel oxazolidinone were synthesized. Antibacterial activity of the compounds against a number of sensitive and resistant bacteria showed promising results. PMID- 19811912 TI - Synthesis of a novel C2/C2'-aryl-substituted pyrrolo[2,1-c][1,4]benzodiazepine dimer prodrug with improved water solubility and reduced DNA reaction rate. AB - A prodrug form (17) of a novel C2/C2'-aryl-substituted pyrrolobenzodiazepine (PBD) dimer (16) has been synthesized by introducing sodium bisulfite groups to the C11/C11'-positions of the parent bis-imine. The prodrug form is highly water soluble, stable in aqueous conditions, and the rate of DNA cross-link formation is much slower compared to the parent bis-imine. PMID- 19811913 TI - Anthranilic sulfonamide CCK1/CCK2 dual receptor antagonists I: discovery of CCKR1 selectivity in a previously CCKR2-selective lead series. AB - A series of CCK2R-selective anthranilic amides is shown to derive CCK1R affinity via selective substitution of the amide side chain. Thus, extending the length of the original benzamide side chain by a single methylene unit imparts CCK1R affinity to the series, and further fine tuning of the affinity results in CCK1R selectivity of greater than 100-fold. PMID- 19811914 TI - Synthesis of modified homo-N-nucleosides from the reactions of mesityl nitrile oxide with 9-allylpurines and their influence on lipid peroxidation and thrombin inhibition. AB - 9-(3-Mesityl-4,5-dihydroisoxazol-5-yl) homo-N-nucleosides were prepared from the 1,3-dipolar cycloaddition reactions of mesityl nitrile oxide with 9-allyl derivatives of 6-chloropurine, 6-piperidinylpurine, 6-morpholinylpurine, 6 pyrrolidinylpurine, and 6-N,N-dibenzoyladenine. The new compounds were tested in vitro for their ability: (i) to interact with 1,1-diphenyl-2-picryl-hydrazyl (DPPH) stable free radical, (ii) to inhibit lipid peroxidation, (iii) to scavenge the superoxide anion, (iv) to inhibit the activity of soybean lipoxygenase, and (v) to inhibit in vitro thrombin. Most of them found to be potent thrombin inhibitors and to inhibit in vitro lipid peroxidation. The majority of the compounds showed significant lipoxygenase inhibitory activity. PMID- 19811915 TI - Pyranocoumarins: a new class of anti-hyperglycemic and anti-dyslipidemic agents. AB - A series of pyranocoumarin derivatives were synthesized and evaluated in vivo for their anti-hyperglycemic as well as anti-dyslipidemic activities. Compounds 7a, 7c, 8a, 8b, 8c, 8e and 8f have shown promising anti-hyperglycemic activities in sucrose loaded model (SLM) as well as sucrose challenged streptozotocin induced diabetic rat model (STZ). Compounds 8a and 8b were showing 38.0% and 42.0% blood glucose lowering activity in db/db mice model. In vitro anti-hyperglycemic activity evaluation exhibited that compounds 8a (IC(50)=24.5 microM) and 8b (IC(50)=36.2 microM) are potential PTP-1B inhibitors thereby revealing their possible mechanism of anti-diabetic action. Compounds 7a, 7b, 8a, 8b, 8d, 8e and 8f have shown significant anti-dyslipidemic activity in triton induced dyslipidemia in rats. PMID- 19811916 TI - Design and synthesis of cell potent BACE-1 inhibitors: structure-activity relationship of P1' substituents. AB - Using structure-guided design, hydroxyethylamine BACE-1 inhibitors were optimized to nanomolar Abeta cellular inhibition with selectivity against cathepsin-D. X ray crystallography illuminated the S1' residues critical to this effort, which culminated in compounds 56 and 57 that exhibited potency and selectivity but poor permeability and high P-gp efflux. PMID- 19811917 TI - Stereoselective total synthesis of a potent natural antifungal compound (6S) 5,6,dihydro-6-[(2R)-2-hydroxy-6-phenyl hexyl]-2H-pyran-2-one. AB - A practical stereoselective synthesis of (6S)-5,6,dihydro-6-[(2R)-2-hydroxy-6 phenyl hexyl]-2H-pyran-2-one, a potent natural antifungal compound, is described. The sequence involves diastereoselective iodine-induced electrophilic cyclization, epoxide ring opening with a vinyl Grignard reagent and ring closing metathesis (RCM) as the key steps. PMID- 19811918 TI - Efficient reduction of loco-regional recurrences but no effect on mortality twenty years after postmastectomy radiation in premenopausal women with stage II breast cancer - a randomized trial from the South Sweden Breast Cancer Group. AB - PURPOSE: To study long term loco-regional and distant recurrence rate and survival after post-mastectomy radiotherapy in combination with oral cyclophosphamide in premenopausal women with stage II breast cancer. STUDY DESIGN: A three-armed randomized multicenter phase III trial comparing 1) Radiotherapy (RT) 2) RT+ oral cyclophosphamide for one year (RT+C) and 3) Oral cyclophosphamide only (C). Radiotherapy was administered, in 20 fractions, to 48Gy to the axilla and parasternal lymph nodes, 45Gy to infra- and supraclavicular fossae and 38Gy to the chest wall. Cyclophosphamide was prescribed as 12 courses of 130mg/m(2) od for 14 days every 4 weeks. PATIENTS AND METHODS: 367 patients from 15 surgical departments in Southern Sweden, representing 80% of all eligible patients, were included in the trial between 1978-1983. Median age was 47 years, median tumour size was 25mm, and 33% of the patients were lymph node negative. Median follow-up time was 24 years. RESULTS: RT reduced the risk at twenty years for loco-regional recurrence in C-treated patients at twenty years with 75% (13.9% vs. 3.5%). The risk reduction was highly significant in both N0 and N+ patients. No reduction in systemic disease or mortality was observed. CONCLUSION: Post-mastectomy radiotherapy reduced loco regional recurrences in this premenopausal population, but no effect was seen on mortality with 20 years follow-up. PMID- 19811919 TI - Incidental walking activity is sufficient to induce time-dependent conditioning of the Achilles tendon. AB - The Achilles tendon has been seen to exhibit time-dependent conditioning when isometric muscle actions were of a prolonged duration, compared to those involved in dynamic activities, such as walking. Since, the effect of short duration muscle activation associated with dynamic activities is yet to be established, the present study aimed to investigate the effect of incidental walking activity on Achilles tendon diametral strain. Eleven healthy male participants refrained from physical activity in excess of the walking required to carry out necessary daily tasks and wore an activity monitor during the 24 h study period. Achilles tendon diametral strain, 2 cm proximal to the calcaneal insertion, was determined from sagittal sonograms. Baseline sonographic examinations were conducted at approximately 08:00 h followed by replicate examinations at 12 and 24 h. Walking activity was measured as either present (1) or absent (0) and a linear weighting function was applied to account for the proximity of walking activity to tendon examination time. Over the course of the day the median (min, max) Achilles tendon diametral strain was -11.4 (4.5, -25.4)%. A statistically significant relationship was evident between walking activity and diametral strain (P<0.01) and this relationship improved when walking activity was temporally weighted (AIC 131 to 126). The results demonstrate that the short yet repetitive loads generated during activities of daily living, such as walking, are sufficient to induce appreciable time-dependant conditioning of the Achilles tendon. Implications arise for the in vivo measurement of Achilles tendon properties and the rehabilitation of tendinopathy. PMID- 19811920 TI - Glutaredoxins: roles in iron homeostasis. AB - Glutaredoxins, proteins traditionally involved in redox reactions, are also required for iron-sulfur cluster assembly and haem biosynthesis. These new roles are probably related to the ability of some glutaredoxins to bind labile [2Fe-2S] clusters and to transfer them rapidly and efficiently to acceptor proteins. Recent results point to putative roles for glutaredoxins in the sensing of cellular iron and in iron-sulfur cluster biogenesis, either as scaffold proteins for the de novo synthesis of iron-sulfur clusters or as carrier proteins for the transfer of preformed iron-sulfur clusters. Based on prokaryote genome analysis and in vivo studies of iron regulation in yeast, we propose putative new roles and binding partners for glutaredoxins in the assembly of metalloproteins. PMID- 19811921 TI - In vivo and in vitro anti-leishmanial activities of 4-nitro-N-pyrimidin- and N pyrazin-2-ylbenzenesulfonamides, and N2-(4-nitrophenyl)-N1-propylglycinamide. AB - A series of compounds containing the nitrobenzene and sulfonamido moieties were synthesized and their leishmanicidal effect was assessed in vitro against Leishmaniainfantum promastigotes. Among the compounds evaluated, the p nitrobenzenesulfonamides 4Aa and 4Ba, and the p-nitroaniline 5 showed significant activity with a good selectivity index. In a Balb/c mice model of L. Infantum, administration of compounds 4Aa, 4Ba or 5 (5mg/kg/day for 10 days, injected ip route) led to a clear-cut parasite burden reduction (ca. 99%). In an attempt to elucidate their mechanism of action, the DNA interaction of 4Aa and 5 was investigated by means of viscosity studies, thermal denaturation and nuclease activity assay. Both compounds showed nuclease activity in the presence of copper salt. The results suggest that compounds 4Aa, 4Ba and 5 represent possible candidates for drug development in the therapeutic control of leishmaniasis. PMID- 19811922 TI - Bioorganic synthesis of end-capped anti-HIV peptides by simultaneous cyanocysteine-mediated cleavages of recombinant proteins. AB - Bioorganic synthesis of N- and C-terminal end-capped peptides by two simultaneous S-cyanocysteine-mediated cleavages of recombinant proteins is described. This approach is demonstrated in the preparation of anti-HIV fusion inhibitory peptides. PMID- 19811923 TI - 5-Nitrofuranes and 5-nitrothiophenes with anti-Trypanosoma cruzi activity and ability to accumulate squalene. AB - Chagas disease represents a serious public health problem in South America. The first line of treatment is Nifurtimox and Benznidazole which generate toxic effects in treated patients. We have recently shown that a number of 5 nitrofuranes possess activity against Trypanosoma cruzi through oxidative stress and inhibition of parasite ergosterol biosynthesis, specifically at the level of squalene epoxidase. Here, we identify new 5-nitrofuranes and the thia-analogues with excellent effects on the viability of T. cruzi and adequate parasite/mammal selectivity indexes. Analysis of the free sterols from parasite incubated, during 120h, with the compounds showed that some of them accumulated squalene suggesting the squalene epoxidase activity inhibition of the parasite. Nifurtimox was able to accumulate squalene only at lower incubation times. Due to this fact some derivatives were also tested as antifungal agents. Quantitative structure activity relationship studies were also performed showing relevant features for further new derivatives design. Taken together, the results obtained in the present work point to a more general effect of 5-nitrofuranes and 5 nitrothiophenes in trypanosomatids, opening potential therapeutic possibilities of them for these infectious diseases. PMID- 19811924 TI - Development of a highly water-soluble peptide-based human neutrophil elastase inhibitor; AE-3763 for treatment of acute organ injury. AB - A series of peptide-based transition-state human neutrophil elastase (HNE) inhibitors with N-terminal acidic moieties were synthesized and their inhibitory activity against HNE was evaluated both in vitro and in vivo. Our results show that compounds containing cyclic amide bridged acidic moieties at the N-terminal have not only improved water solubility but also high in vivo potency. Among these compounds, AE-3763 showed remarkable efficacy in hamster models of elastase induced lung hemorrhage and lipopolysaccharide (LPS)-induced lung injury as well as in a mouse model of LPS/galactosamine-induced acute multiple organ dysfunctions. The water solubility of AE-3763 (>1000 mg/ml in H(2)O) was also far superior to that of any of the other compounds synthesized. Thus, it is believed that AE-3763 would be useful for treatment of HNE-associated respiratory disorders, such as acute respiratory distress syndrome (ARDS), acute lung injury (ALI), and acute exacerbation of chronic obstructive pulmonary disease (COPD). PMID- 19811925 TI - Cross section measurement for the (95)Mo(n, alpha)(92)Zr reaction at 4.0, 5.0 and 6.0MeV. AB - Measurements of cross sections of the (95)Mo(n, alpha)(92)Zr reaction at E(n)=4.0, 5.0 and 6.0MeV were carried out at the 4.5MV Van de Graaff of Peking University, China. A twin gridded ionization chamber and two large-area (95)Mo samples were adopted. Fast neutrons were produced through the D(d, n)(3)He reaction by using a deuterium gas target. A small (238)U fission chamber was employed for absolute neutron flux determination. Present data are compared with existing evaluations and measurement. PMID- 19811926 TI - Functional characterization of three water deficit stress-induced genes in tobacco and Arabidopsis: an approach based on gene down regulation. AB - Functional characterization of water deficit stress responsive genes is important to understand their role in stress tolerance. RNAi-based silencing of gene of interest and studying the stress response of knockdown plants under stress can be one of the potential options for assessing functional significance of these genes. Several genes showing higher transcript expression under water deficit stress were cloned earlier from a stress adapted crop species, groundnut. In this study, a few selected gene homologs have been characterized in Nicotiana tabacum and Arabidopsis. Using post transcriptional gene silencing (PTGS) based RNAi approach we developed N. tabacum knockdown lines for three of the genes namely alcohol dehydrogenase (ADH), trans caffeoyl coA-3-O-methyl transferase (CcoAOMT) and flavonol-3-O-glucosyl transferase (F3OGT). By quantitative RT-PCR we demonstrated that the RNAi lines showed significant reduction in target gene transcripts. We followed a stress imposition protocol that allows the plants to experience initial gradual acclimation stress and subsequently severe stress for a definite period. The RNAi knockdown lines generated against ADH and F3OGT, when subjected to water deficit stress showed susceptible symptoms signifying the relevance of these genes under stress. Knockdown of CcoAOMT showed higher chlorophyll degradation and less cell viability upon stress compared to control plants. Further, the Arabidopsis mutant lines clearly showed susceptibility to salinity and water deficit stresses validating relevance of these three genes under abiotic stresses. PMID- 19811927 TI - Visceral protection during moderately hypothermic selective antegrade cerebral perfusion through right brachial artery. AB - OBJECTIVE: This study investigates distal organ protection during moderately hypothermic right brachial artery selective antegrade cerebral perfusion (MHSACP) in patients undergoing aortic arch repair. METHODS: Fifty patients undergoing ascending aorta/aortic arch repair using the MHSACP technique (study group) were compared with another 50 patients randomly selected amongst those undergoing moderately hypothermic open heart procedures (control group). The data from successive clinical examinations and blood samplings were then compared to detect any sign of distal organ damage attributable to the MHSACP technique. RESULTS: The mortality rate did not differ between the two groups (2%). The average cardiopulmonary bypass and aortic cross-clamping durations were 154+/-53 min versus 102+/-25 min and 101+/-41 min versus 70+/-31 min for the study and control groups, respectively. The mean MHSACP period was 32+/-14 min (11-81 min) in the study group. The average discharge time was 6.4+/-1.4 days (5-12 days). Repeated clinical examinations revealed no organ dysfunction and there was no difference between the two groups (with the exception of lactate dehydrogenase) with regard to compared biochemical markers, which might be suggestive of a visceral damaging effect of the MHSACP technique. CONCLUSIONS: The present data show that with the use of MHSACP, distal organs are relatively well protected during aortic arch repairs. PMID- 19811928 TI - Combined effects of laser-ICG photothermotherapy and doxorubicin chemotherapy on ovarian cancer cells. AB - Doxorubicin (DOX) is an anthracycline antibiotic widely used in cancer chemotherapy. Its use is limited by cardiac toxicity and drug resistance. Hyperthermia can aid the functionality of DOX, but current hyperthermia delivery methods are hard to apply selectively and locally. The slow temperature increase associated with the external heating may lead to thermal tolerance in cancer cells. The FDA approved dye indocynine green (ICG) has been demonstrated to absorb near-infrared (NIR) light at 808 nm (ideal for tissue penetration) and emit the energy as heat, making it an ideal agent for localized hyperthermia with a rapid rate of temperature increase. The purpose of this study was to investigate the in vitro cytotoxic effect of combined chemotherapy and hyperthermia to a DOX resistant ovarian cancer cell line (SKOV-3). The effect of two different heating methods, ICG induced rapid rate heating and an incubator induced slow rate heating, were compared. All the experiments were conducted in 96-well plates. Cells were subjected to different concentrations of DOX and 60 min 43 degrees C incubation or 5 microM of ICG with 1 min 808 nm NIR laser. SRB assay was used to measure cell proliferation. ICG itself without laser irradiation was not toxic to SKOV-3 cells. The two types of hyperthermia individually produced similar cytotoxicity. DOX by itself was toxic with an IC(50) value of about 5 microM. Hyperthermia in combination with DOX achieved significantly greater cell killing/growth inhibition at all DOX concentrations compared to DOX alone. A subadditive cytotoxic effect was observed by combining DOX and 60 min 43 degrees C incubation which lead to a lowered DOX IC(50) value of about 1 microM. This value was even lower with 1 min laser-ICG photothermotherapy (0.1 microM) and, though not statistically significant, a synergistic effect may exist between DOX and laser-ICG photothermotherapy. The rate of heating may have an effect on chemotherapy-hyperthermia interaction. In conclusion, the combination of photothermal therapy and chemotherapy may provide a valuable tool for cancer treatment with minimized side effect. PMID- 19811929 TI - Glucose transporters in gametes and preimplantation embryos. AB - The oocyte, sperm and preimplantation embryo have unique metabolic needs that must be met to ensure successful pregnancy. The family of facilitative glucose transporters (GLUTs) plays a major role in providing metabolic substrates to these tissues. The variety of GLUTs expressed in these tissues allows for flexibility to adapt to a changing environment. Alterations in glucose transport and metabolism at the earliest stages of development can impact fetal development. Research into the mechanisms of normal glucose transport into cells is critical for improving outcomes in the increasingly common diabetic maternal environment. Here, we review the current understanding in the distribution and role of glucose transporters in gametes and preimplantation embryos under normal and diabetic conditions. PMID- 19811930 TI - Electron capture dissociation product ion abundances at the X amino acid in RAAAA X-AAAAK peptides correlate with amino acid polarity and radical stability. AB - We present mechanistic studies aimed at improving the understanding of the product ion formation rules in electron capture dissociation (ECD) of peptides and proteins in Fourier transform ion cyclotron resonance mass spectrometry. In particular, we attempted to quantify the recently reported general correlation of ECD product ion abundance (PIA) with amino acid hydrophobicity. The results obtained on a series of model H-RAAAAXAAAAK-OH peptides confirm a direct correlation of ECD PIA with X amino acid hydrophobicity and polarity. The correlation factor (R) exceeds 0.9 for 12 amino acids (Ile, Val, His, Asn, Asp, Glu, Gln, Ser, Thr, Gly, Cys, and Ala). The deviation of ECD PIA for seven outliers (Pro is not taken into consideration) is explained by their specific radical stabilization properties (Phe, Trp, Tyr, Met, and Leu) and amino acid basicity (Lys, Arg). Phosphorylation of Ser, Thr, and Tyr decreases the efficiency of ECD around phosphorylated residues, as expected. The systematic arrangement of amino acids reported here indicates a possible route toward development of a predictive model for quantitative electron capture/transfer dissociation tandem mass spectrometry, with possible applications in proteomics. PMID- 19811931 TI - Influence of the organic layer thickness in (metal-assisted) secondary ion mass spectrometry using Ga+ and C60+ projectiles. AB - This article investigates the influence of the organic film thickness on the characteristic and molecular ion yields of polystyrene (PS), in combination with two different substrates (Si, Au) or gold condensation (MetA-SIMS), and for atomic (Ga+) and polyatomic (C60+) projectile bombardment. PS oligomer (m/z approximately 2000 Da) layers were prepared with various thicknesses ranging from 1 up to 45 nm on both substrates. Pristine samples on Si were also metallized by evaporating gold with three different thicknesses (0.5, 2, and 6 nm). Secondary ion mass spectrometry was performed using 12 keV atomic Ga+ and C60+ projectiles. The results show that upon Ga+ bombardment, the yield of the fingerprint fragment C7H7+ increases as the PS coverage increases and reaches its maximum for a thickness that corresponds to a complete monolayer (approximately 3.5 nm). Beyond the maximum, the yields decrease strongly and become constant for layers thicker than 12 nm. In contrast, upon C60+ bombardment, the C7H7+ yields increase up to the monolayer coverage and they remain constant for higher thicknesses. A strong yield enhancement is confirmed upon Ga+ analysis of gold-metallized layers but yields decrease continuously with the gold coverage for C60+ bombardment. Upon Ga+ bombardment, the maximum PS fingerprint ion yields are obtained using a monolayer spin-coated on gold, whereas for C60+, the best results are obtained with at least one monolayer, irrespective of the substrate and without any other treatment. The different behaviors are tentatively explained by arguments involving the different energy deposition mechanisms of both projectiles. PMID- 19811933 TI - Multiple imputation for missing laboratory data: an example from infectious disease epidemiology. AB - PURPOSE: To present multiple imputation (MI) as an appropriate method to address missing values for a laboratory parameter (serum albumin) in an epidemiologic study. METHODS: A data set of patients who were hospitalized for invasive group A streptococcal infections was accessed. Age was the exposure of interest. The outcome was hospital mortality. Several variables, including serum albumin, were considered to be potential confounders. Of the 201 records, 91 had missing values for serum albumin. The MI procedure in SAS was used to perform 20 imputations of serum albumin by using a Markov chain Monte Carlo approach. Logistic regression was then performed on each of the 20 filled-in data sets, and the results were appropriately combined by using the MIANALYZE procedure. RESULTS: Age (> or = 55 years vs. 0-54 years) was not a risk factor for hospital mortality in the complete-case analysis (n = 110): adjusted odds ratio (OR) = 2.43 (95% confidence interval [CI]: 0.79-7.53). Age was a significant risk factor in the imputed data set (n = 201): adjusted OR = 3.08 (95% CI: 1.22-7.78). CONCLUSIONS: Epidemiologists frequently encounter data sets that contain missing values. Traditional missing data techniques such as the complete-subject analysis may lead to biased results. We have demonstrated the use of a novel technique, MI, to account for missing data. PMID- 19811932 TI - Identification of alpha- and beta-hydroxy acid containing cyclodepsipeptides in natural peptide mixtures using negative ion mass spectrometry. AB - Natural peptide libraries often contain cyclodepsipeptides containing alpha- or beta-hydroxy residues. Extracts of fungal hyphae of Isaria yield a microheterogenous cyclodepsipeptide mixture in which two classes of molecules can be identified by mass spectral fragmentation of negative ions. In the case of isaridins, which contain an alpha-hydroxy residue and a beta-amino acid residue, a characteristic product ion corresponding to a neutral loss of 72 Da is obtained. In addition, neutral loss of water followed by a 72 Da loss is also observed. Two distinct modes of fragmentation rationalize the observed product ion distribution. The neutral loss of 72 Da has also been obtained for a roseotoxin component, which is also an alpha-hydroxy residue containing cyclodepsipeptide. In the case of isariins, which contain a beta-hydroxy acid residue, ring opening and subsequent loss of the terminal residue as an unsaturated ketene fragment, rationalizes the observed product ion formation. Fragmentation of negative ions provide characteristic neutral losses, which are diagnostic of the presence of alpha-hydroxy or beta-hydroxy residues. PMID- 19811934 TI - Shoulder and elbow muscle activity during fully supported trajectory tracking in people who have had a stroke. AB - An inability to perform tasks involving reaching is a common problem for stroke patients. This paper provides an insight into mechanisms associated with recovery of upper limb function by examining how stroke participants' upper limb muscle activation patterns differ from those of neurologically intact participants, and how they change in response to an intervention. In this study, five chronic stroke participants undertook nine tracking tasks in which trajectory (orientation and length), speed and resistance to movement were varied. During these tasks, EMG signals were recorded from triceps, biceps, anterior deltoid, upper, middle and lower trapezius and pectoralis major. Data collection was performed in sessions both before, and after, an intervention in which participants performed a similar range of tracking tasks with the addition of responsive electrical stimulation applied to their triceps muscle. The intervention consisted of eighteen one hour treatment sessions, with two participants attending an additional seven sessions. During all sessions, each participant's arm was supported by a hinged arm-holder which constrained their hand to move in a two dimensional plane. Analysis of the pre intervention EMG data showed that timing and amplitude of peak EMG activity for all stroke participants differed from neurologically intact participants. Analysis of post intervention EMG data revealed that statistically significant changes in these quantities had occurred towards those of neurologically intact participants. PMID- 19811935 TI - Types of attention matter for awareness: a study with color afterimages. AB - It has been argued that attention and awareness might oppose each other given that attending to an adapting stimulus weakens its afterimage. We argue instead that the type of attention guided by spatial extent and perceptual levels is critical and might result in differences in awareness using afterimages. Participants performed a central task with small, large, local, or global letters and a blue square as an adapting stimulus in three experiments and indicated the onset and offset of the afterimage. We found that increases in the spatial spread of attention resulted in the decrease of afterimage duration. In terms of levels of processing, global processing produced larger afterimage durations with stimuli controlled for spatial extent. The results suggest that focused or distributed attention produce different effects on awareness, possibly through their differential interactions with polarity dependent and independent processes involved in the formation of color afterimages. PMID- 19811936 TI - Term intra-partum asphyxia: an analysis of acute non-specific supportive criteria and non-CNS organ injury. AB - OBJECTIVE: The purpose of this study was to describe the frequencies and relationships of non-specific non-essential diagnostic criteria and non-CNS organ system injury in term intra-partum asphyxia. METHODS: All children with term intra-partum asphyxia encountered in a single pediatric neurology practice with at least two years follow-up and an abnormal neurologic outcome were identified. RESULTS: A total of 40 children (28 males, 12 females) were identified. Twenty four had moderate NE and sixteen severe NE. The mean number of non-specific non essential diagnostic criteria (out of a possible 7) was 4.75+/-1.39 SD. Sixty percent had five or more criteria and all criteria were present in only 10% of newborns. The mean number of non-CNS organ systems affected was 2.88+/-1.96 SD (out of a possible 6). Ten percent of our sample showed no evident non-CNS organ injury acutely. CONCLUSION: Most asphyxiated neonates failed to consistently satisfy all elements of present consensus statements. PMID- 19811937 TI - Low back pain may be caused by disturbed pain regulation: a cross-sectional study in low back pain patients using tender point examination. AB - BACKGROUND: Widespread pain has negative influence on outcome in low back pain (LBP) patients. Tender point (TP) examination is a standardized examination method to estimate diffuse tenderness. AIMS: To assess diffuse tenderness by means of a standardized TP examination and to analyse for associations between the number of TPs and spinal structural changes as well as psycho-social factors. METHODS: Patients sick-listed 3-16 weeks due to LBP with or without sciatica completed a questionnaire and went through a clinical low back examination and TP examination. Of 326 patients 111 had verified nerve root affection and 215 had non-specific LBP with or without radiating pain. Disc height reductions were estimated on lateral X-rays. RESULTS: Multivariate logistic regression analysis showed that more than 8 TPs were strongly negatively associated with disc degeneration (Odds Ratio (OR) 0.58 (0.40-84), 95% Confidence Interval (CI): 0.39 0.84, p=0.004) and verified nerve root affection (OR 0.15 (0.04-0.54), p=0.004) and were positively associated with number of years since first episode of LBP (OR 1.05, CI: 1.01-1.09, p=0.009). Furthermore, more than 8 TPs were positively associated with widespread pain, female sex and bodily distress. With all patients included, bodily distress and the number of tender points were positively associated with the intensity of LBP, but disc degeneration was only positively associated with LBP in patients with less than 6 TPs. CONCLUSIONS: The pain in patients with diffuse tenderness was rarely related to disc degeneration or nerve root affection, rather it may be caused by disturbed pain regulation. PMID- 19811938 TI - Cross-sectional survey of Toxoplasma gondii infection in colony cats from urban Florence (Italy). AB - Cats are the key species in the epidemiology of Toxoplasma gondii infection, even if the proportion of subjects excreting oocysts is low. The aim of the present paper was to obtain information about seroprevalence, oocyst shedding rate and presence of T gondii DNA in faeces collected from an urban population of colony cats in Florence (Tuscany). Fifty European shorthair feral cats were examined for anti-T gondii specific antibodies by a modified agglutination test (MAT), and for oocysts by microscopic examination and for faecal protozoal DNA, by means of a nested polymerase chain reaction (n-PCR) protocol. Twenty-two out of 50 serum samples (44%) were MAT positive. T gondii oocysts were not detected in any of the examined faecal samples. Eight out of 50 faecal specimens (16%) were n-PCR positive and sequencing of the bands was specific for T gondii. Detection by combination of the two methods was higher than single techniques and enhanced the detection of T gondii up to 48%. Our results suggest that the use of MAT plus PCR in faeces may be the best choice for diagnosis of feline toxoplasmosis. Further studies to ascertain the real infectivity of the copro-PCR positive subjects are required. PMID- 19811939 TI - Comparative study of complete versus incomplete Kawasaki disease in 59 pediatric patients. AB - OBJECTIVES: To compare the clinical and laboratory features and the rate of echocardiographic coronary artery abnormalities in patients with complete and incomplete forms of Kawasaki disease (KD) and to determine which additional clinical criteria might support a suspicion of KD. METHODS: We retrospectively reviewed the medical records of patients with KD who were admitted to the general pediatrics department of the Kremlin Bicetre Teaching Hospital, France, between January 1995 and May 2006. We compared patients with a fever and four or five of the principal criteria (complete KD) to the other patients (incomplete KD). Clinical and laboratory features were abstracted from the records. RESULTS: We identified 63 patients with a mean age of 33 months (+/-31). The male-to-female ratio was 2.47. Four patients were excluded. Of the remaining 59 patients, 39 had complete KD and 20 incomplete KD. The group with complete KD had significantly higher rates of changes in the extremities, conjunctival injection, exanthem, and enanthem; and a significantly lower rate of coronary artery dilation (48.7% vs. 90% in the incomplete KD group, P=0.002). Serum levels of alanine aminotransferase and gamma glutamyl transferase were significantly higher in the complete KD group. No significant differences were found between the two groups regarding age, sex, blood cell counts, or laboratory markers for inflammation. Pyuria was found in 45.4% of patients with complete KD and in 30.8% of those with incomplete KD (P=0.17). Of 14 patients who underwent ophthalmological evaluation, two had uveitis; both of them had complete KD. CONCLUSION: Incomplete KD shares with complete KD a risk of coronary artery disease. The diagnosis of incomplete KD is challenging but can be supported by the presence of features other than the principal criteria, such as acute anterior uveitis or unexplained pyuria. PMID- 19811940 TI - [Hand and foot acrometastasis secondary to breast carcinoma]. AB - Hands and feet acrometastases are rarely observed. The authors report a case of bone metastasis to the left thumb and the right second metatarsal bone in a 37 year-old woman with breast carcinoma. Through this case report, mechanisms of such metastases are reviewed. PMID- 19811941 TI - [Spinal migration of a Kirschner wire after surgery for clavicular nonunion. A case report and review of the literature]. AB - Kirschner wires (K-wires) are often used for osteosynthesis particularly in the upper limb. Postoperative K-wire migration through the tissues is a well recognised and significant complication of surgery of the clavicle, the wire ending up in the lungs, the oesophagus, the aorta, or the subclavian artery. Localisation of a K-wire migration into the spinal cord is very rare. We report the case of a 34-year-old man with K-wire migration into the spinal cord through the intervertebral foramen of T2, two months after surgery for nonunion of a fracture of the lateral clavicle. Apart from acute respiratory failure related to a pneumothorax, the patient initially had no neurological deficit. It was decided to operate on him immediately. Two therapeutic options are possible: simple K wire removal via a supraclavicular approach, or the same but with direct visual control in the spinal cord after laminectomy. A postoperative check with an MRI scan is desirable after two days. Even if mechanism of K-wire migration is not known, the means of prevention are, namely strict postoperative immobilization, K wire removal as soon as bone healing is achieved, and bending the external tip of each implanted wire. PMID- 19811942 TI - [Articular coronal fractures of the distal humerus]. AB - INTRODUCTION: The vague term of capitellar fractures is still frequently used to designate articular coronal fractures of the distal humeral epiphysis. The use of eponyms for their descriptions may cause confusion. Recent publications describe a wide variety of fracture types and recommend new classifications based on the operative findings. We report our results of surgical treatment of 12 cases of these fractures in comparison to recent series of the literature. METHODS: Twelve patients (seven female and five male with a mean age of 31 years and 6 months) have been treated for articular coronal fractures of the distal humeral epiphysis between 1994 and 2004. A retrospective analysis of the radiographs and the operative notes permits their differentiation into 3 types according to the classification of Dubberley et al. (2006): ten fractures of type 1, one fracture of type 2 and one fracture of type 3. All fractures underwent open reduction and internal fixation, except for one case, which was initially missed and operated, therefore, by excision of the articular fragment with a delay of 6 weeks. All patients were clinically evaluated according to the index of performance of Morrey et al. (1993). In addition, a radiological assessment based on the scale of Knirk and Jupiter (1986) for elbow osteoarthritis was performed. RESULTS: The mean follow-up was 9 years. The clinical evaluation showed seven excellent results (six cases of type 1 and one case of type 3 with a score of 100 points for each one), two good (type 1 with 80 and 85 points of respective scores) and three fair (two cases of type 1 with 65 and 60 points of respective scores and one case of type 2 with a score of 65 points). The radiological evaluation showed seven elbows of grade 0 (six cases of type 1 and one case of type 3), four elbows of grade 1 (type 1) and one elbow of grade 2 (type 2). DISCUSSION: Articular coronal fractures of the distal humerus are rare. The classification of Dubberley et al. (2006) [7] is comprehensive and allows inclusion of all varieties of these fractures. In addition, it is the only one that indicates the surgical approach according to the fracture type. However, to do so, a preoperative CT-scan is highly recommended. The more the fracture line extends medially to involve the trochlea (types 2 and 3), the less a lateral approach is sufficient and the more a combined lateral and medial or a posterior transolecranon approach is mandatory. An internal fixation using conventional small fragment screws inserted from posterior to anterior is feasible when the articular fragment has a sufficient subchondral bone thickness. A direct anteroposterior fixation is better achieved using headless screws buried beneath the cartilaginous surface; it is particularly helpful when the articular fragment has a thin sub-chondral cancellous bone component. Excision is reserved for comminuted fractures, those not amenable to fixation, very thin or osteoporotic fragments, and for the late diagnosed fracture. PMID- 19811943 TI - Bubble population phenomena in sonochemical reactor: I estimation of bubble size distribution and its number density with pulsed sonication - laser diffraction method. AB - To characterize the bubble populations (size and its number distribution) in a sonochemical reactor, a simple but powerful technique based on the Fraunhofer laser diffraction (LD) has been proposed. In this method, the acoustic wave disturbance to the laser probe in the sonochemical reaction field was eliminated by the temporal separation using pulsed sonication (pulsed LD). With this relatively simple strategy, the temporal development of the bubble size distribution could be evaluated by pulsed LD. A number density of bubbles was estimated by using a calibration data obtained with monosized standard particles. In addition, the effect of pulse length and a surfactant on the bubble population phenomena in a multibubble system are discussed. PMID- 19811944 TI - The safety of transcranial magnetic stimulation with deep brain stimulation instruments. AB - OBJECTIVES: Transcranial magnetic stimulation (TMS) has been employed in patients with an implanted deep brain stimulation (DBS) device. We investigated the safety of TMS using simulation models with an implanted DBS device. METHODS: The DBS lead was inserted into plastic phantoms filled with dilute gelatin showing impedance similar to that of human brain. TMS was performed with three different types of magnetic coil. During TMS (1) electrode movement, (2) temperature change around the lead, and (3) TMS-induced current in various situations were observed. The amplitude and area of each evoked current were measured to calculate charge density of the evoked current. RESULTS: There was no movement or temperature increase during 0.2 Hz repetitive TMS with 100% stimulus intensity for 1 h. The size of evoked current linearly increased with TMS intensity. The maximum charge density exceeded the safety limit of 30 muC/cm(2)/phase during stimulation above the loops of the lead with intensity over 50% using a figure-eight coil. CONCLUSIONS: Strong TMS on the looped DBS leads should not be administered to avoid electrical tissue injury. Subcutaneous lead position should be paid enough attention for forthcoming situations during surgery. PMID- 19811946 TI - Current insights into phage biodiversity and biogeography. AB - Phages exert tremendous ecological and evolutionary forces directly on their bacterial hosts. Phage induced cell lysis also indirectly contributes to organic and inorganic nutrient recycling. Phage abundance, diversity, and distribution are therefore important parameters in ecosystem function. The assumption that phage consortia are ubiquitous and homogenous across habitats (everything is everywhere) is currently being re-evaluated. New studies on phage biogeography have found that some phages are globally distributed while others are unique and perhaps endemic to specific environments. Furthermore, advances in technology have allowed scientists to conduct experiments aimed at analyzing phage consortia over temporal scales, and surprisingly have found reoccurring patterns. This review discusses currents in the field of phage ecology with particular focus on efforts to characterize phage diversity and biogeography across various spatial and temporal scales. PMID- 19811945 TI - Caring for Machado-Joseph disease: current understanding and how to help patients. AB - Machado-Joseph disease or spinocerebellar ataxia 3 (MJD/SCA3) is a clinically heterogeneous, neurodegenerative disorder characterized by varying degrees of ataxia, ophthalmoplegia, peripheral neuropathy, pyramidal dysfunction and movement disorder. MJD/SCA3 is caused by a CAG repeat expansion mutation in the protein coding region of the ATXN3 gene located at chromosome 14q32.1. Current hypotheses regarding pathogenesis favor the view that mutated ataxin-3, with its polyglutamine expansion, is prone to adopt an abnormal conformation, engage in altered protein-protein interactions and aggregate. Expanded CAG repeat length correlates with the range and severity of the clinical manifestations and inversely correlates with age of disease onset. Though MJD/SCA3 is classically described as affecting the cerebellum, brainstem and basal ganglia, recent neuropathology and neuroimaging series demonstrate involvement of other areas such as the thalamus and cerebral cortex. Clinically, much emphasis has been placed in the description and recognition of the non-motor symptoms observed in these patients, such as pain, cramps, fatigue and depression. Currently, no disease modifying treatment exists for MJD/SCA3. Standard of care includes genetic counseling, exercise/physical therapy programs, and speech and swallow evaluation. Symptomatic treatment for clinical findings such as depression, sleep disorders, parkinsonism, dystonia, cramps, and pain is important to improve the quality of life for those with MJD/SCA3. PMID- 19811947 TI - Does hygiene counseling have an impact on the rate of CMV primary infection during pregnancy? Results of a 3-year prospective study in a French hospital. AB - BACKGROUND: Cytomegalovirus (CMV) is the most frequent cause of congenital viral infection in developed countries. OBJECTIVES: The objective of this study was to evaluate the impact of our prenatal CMV infection screening and counseling policy. STUDY DESIGN: Since 2005, all pregnant women in our obstetric center have been informed about CMV infection, and if they agree, given a serological test at around 12 weeks of gestation (WG). If this first test is negative, the women and their partners are given hygiene counseling on how to prevent CMV infection, and a second test is performed at around 36 WG. RESULTS: Among the 5312 women who had an unknown immune status, or were known to be seronegative when they had their first visit to our center for their current pregnancy, 97.4% agreed to CMV screening. Primary infection was detected in 11 women between 0 and 12 WG (0.42%), and seroconversion was diagnosed in five women between 12 and 36 WG (0.19%). CONCLUSIONS: These results suggest that if clear information is given on CMV infection during pregnancy, the rate of seroconversion is lower following counseling than before counseling. PMID- 19811948 TI - Genomic variation and evolution of Staphylococcus aureus. AB - The evolution of new human and animal pathogenic strains of Staphylococcus aureus has been due to the accumulation of mobile genetic elements (MGE) encoding methicillin resistance and virulence factors into successful lineages. These include epidemic methicillin-resistant S. aureus in hospitals (EMRSA), community associated MRSA (CA-MRSA), fully vancomycin-resistant MRSA (VRSA) and livestock associated MRSA (LA-MRSA). The S. aureus population in humans is dominated by about ten S. aureus lineages while animals generally have different lineages. Individual isolates within each lineage have unique combination of MGE often encoding virulence and resistance genes. S. aureus evolves due to point mutation and selection, but also dramatically due to the horizontal transfer of these MGE between strains or from other species or genera. Horizontal transfer, by conjugation or transduction, can be blocked by S. aureus restriction modification systems which are lineage specific. Because of the mobility of MGE, there are prospects for increasingly virulent and resistant strains to emerge that could severely affect healthcare and agriculture more effectively than the current pathogens. PMID- 19811949 TI - Strength increases in upper and lower body are larger with longer inter-set rest intervals in trained men. AB - The purpose of the current study was to compare different rest interval durations on upper and lower body strength. Thirty-six recreationally trained men were randomly assigned to 1 min (G1; n=12), 3 min (G3; n=12) or 5 min (G5; n=12) rest interval groups. Each group performed the same resistance training program. Maximal strength was assessed at baseline, mid-point (8 weeks) and post-training (16 weeks) for the bench press and leg press exercises. For the bench press, significant increases were demonstrated within G3 and G5 at 8 weeks and at 16 weeks versus baseline (p<0.05). Additionally, for the bench press, G5 (98.2+/-3.7 kg) was significantly stronger than G1 (92.5+/-3.8 kg) at 16 weeks (p<0.05). For the leg press, significant increases were demonstrated within all groups at 8 weeks and at 16 weeks versus baseline (p<0.05). Additionally, for the leg press, G5 (290.8+/-23.5 kg) was significantly stronger than G1 (251.0+/-15.8 kg) at 8 weeks (p<0.01) and G3 (305.0+/-23.9 kg) and G5 (321.7+/-21.7 kg) were significantly stronger than G1 (276.7+/-10.7 kg) at 16 weeks (p<0.05). The findings of the current study indicate that utilising 3 or 5 min rest intervals between sets may result in significantly greater increases in upper and lower body strength beyond the initial weeks of training versus utilising 1-min rest intervals between sets. PMID- 19811950 TI - Penile length shortening after radical prostatectomy: men's responses. AB - BACKGROUND: Prostate cancer (PC) is the most common type of male-specific cancer in North American men, and many men choose radical prostatectomy (RP) to remove their cancer. Although penile length shortening (PLS) occurs in a reported 68% to 71% of men undergoing RP, little is known about it. In an electronic journal search, only 9 medical articles (with no nursing publications) were published between 1980 and 2007. PURPOSE: To provide an account of patients' perceptions and responses to living with PLS after RP. METHODS AND SAMPLE: Semi-structured interviews and a grounded theory approach were used to discover the basic social processes regarding men's perceptions of a shortened penis and overall sense of self. A total of six men who underwent RP and consequently noticed PLS were recruited from a local PC support group for semi-structured interviews lasting between 40-60 min. RESULTS: Based on subjects' own definitions of masculinity, no significant changes in the constructs of masculinity and overall self-image perception were reported. CONCLUSIONS: Men undergoing RP may not be fully aware that PLS is a possible consequence related to treatment. In spite of this, subjects were not negatively affected by its occurrence. PMID- 19811951 TI - Irritable bowel syndrome: towards biomarker identification. AB - Irritable bowel syndrome (IBS), the most common functional gastrointestinal disorder referred to gastroenterologists, affects 7-10% of the general population worldwide. The lack of suitable disease-defining biological markers coupled with a poorly understood underlying pathophysiology complicates patient diagnosis and seriously hampers drug discovery efforts. Over the past few years, a number of potential biomarkers have emerged, and in this review we critically evaluate such candidates. In particular, we highlight the increasing number of studies supporting a low-grade immune activation in IBS and consider how the latest preclinical developments can contribute to the development of more robust and reliable biological markers of this disorder. The successful identification of biomarkers is critical to progressing our understanding of IBS and addressing the unmet therapeutic needs of this debilitating condition. PMID- 19811952 TI - Oxidative stress in bone remodelling and disease. AB - Oxidative stress is characterised by an increased level of reactive oxygen species (ROS) that disrupts the intracellular reduction-oxidation (redox) balance. Although initially shown to be involved in aging, physiological roles for ROS in regulating cell functions and mediating intracellular signals have emerged. In bone tissues, recent studies have demonstrated that ROS generation is a key modulator of bone cell function and that oxidative status influences the pathophysiology of mineralised tissues. Here, we review the crucial role of oxidative stress in bone pathophysiology, and discuss the possibility that ROS production might be a relevant therapeutic target under certain conditions. Further studies will be needed to investigate whether manipulation of the redox balance in bone cells represents a useful approach in the design of future therapies for bone diseases. PMID- 19811953 TI - Translation of the Nursing Clinical Facilitators Questionnaire (NCFQ) to Norwegian language. AB - The translation and adaptation of English instruments to be used with populations speaking other languages is an important and complex process which is attracting increased attention in nursing and health-related research. The aim of this article is to describe the translation process of the Nursing Clinical Facilitators Questionnaire (NCFQ) for testing in Norway. The instrument is a 28 item-questionnaire with a Likert-type (1-5) scale ranging from the descriptions "strongly agree (1) to strongly disagree (5)". The aim of the instrument is to measure the efficiency of, and satisfaction with the supervision received from the nurse students' perspective. The NCFQ questionnaire was translated in six phases. The translation process was conducted systematically by applying the three methods described in the literature: the methods of forward-translation, back-translation and comparison followed by an empirical study (pilot test). The methods were chosen to test the quality of translation, establish semantic equivalence of the translated instrument and to estimate the cross-cultural relevance of the instrument. The translation process has given prerequisites to use the NCFQ questionnaire in a larger study and a possibility to compare different models for supervision of nursing students in the clinical part of their education. PMID- 19811954 TI - Modulation of the colonic epithelial cell responses and amelioration of inflammation by CD80 blockade in TNBS colitis. AB - Inflammatory bowel diseases (IBD) result from dysregulated immune responses to the luminal antigens initiated by the colonic epithelial cells (CEC) and propagated by activated CD4+ T cells. Biological therapies are being developed that suppress inflammation and promote epithelial homeostasis. Treatment with the CD80-competitive antagonist peptide (CD80-CAP) has been shown to suppress T cell responses in multiple disease models. Here we investigated the effect of the CD80 CAP on the CEC responses in experimental colitis. Balb/c mice induced with trinitrobenzene sulfonic acid (TNBS) colitis were administered CD80-CAP/control peptide/vehicle. Administration of the CD80-CAP decreased microscopic inflammation and restored the expression of TLR-2, 3, 4 and 5 mRNA in the CEC of colitis mice to physiological levels. Furthermore, the CD80-CAP treatment suppressed Th1 cytokines and enhanced Th2 responses by the CEC in colitis mice. In conclusion, CD80-CAP administration ameliorated TNBS colitis by reducing the inflammatory cell infiltration and modulating the CEC response potentially restoring mucosal tolerance. PMID- 19811955 TI - Detection of foetal growth restriction using third trimester ultrasound. AB - Foetal growth restriction is an important contributor to perinatal mortality, being responsible for up to 50% of stillbirths. Optimal prevention and accurate detection enabling timely intervention remain elusive, particularly in presumed low-risk pregnancy. Third trimester ultrasound seems a logical solution, but systematic review of evidence from randomised trials has shown that third trimester ultrasound does not have a significant impact on perinatal mortality but may increase interventions such as caesarean delivery. However, the evidence is difficult to interpret in the context of current obstetric practice as the evolution of ultrasound technology and rapid assimilation of newer techniques has resulted in questionable validity of the findings. If third trimester ultrasound were introduced routinely, there is a need to decide the optimal timing and number of examinations and what ultrasound parameters should be used to identify the foetus at risk. PMID- 19811956 TI - Milestones in the management of atrial fibrillation. AB - Over the past decades, interest in atrial fibrillation (AF) has greatly increased, and the understanding of its pathophysiology and potential treatment modalities is constantly growing. This article summarizes findings that the authors deem milestones in the clinical management of the arrhythmia. The first milestone was the observation that AF is not a benign entity; rather, it is associated with an increased risk of death and morbidity. While no trial had previously shown that patients live longer if AF is suppressed, the epidemiological association is very consistent among several populations. The second milestone was the discovery that thromboembolic strokes can be prevented by warfarin or (to a lesser extent) antithrombotic therapy. The third milestone was the finding that-with contemporary treatment-rhythm control did not improve patient outcomes and, in fact, that with regard to mortality, rate- and rhythm control strategies are largely interchangeable. Fourth, the description of a specific trigger of the arrhythmia localized in the pulmonary veins has driven efforts to curatively treat AF. While no survival benefit has so far been shown for patients treated with catheter ablation, this technique has progressed from an innovative investigational procedure to routine clinical practice. The fifth milestone is the addition of outcome data to the current body of evidence. In the ATHENA trial, for the first time, an antiarrhythmic agent proved to reduce cardiovascular mortality and morbidity. In summary, the development of novel treatment strategies for AF is highly dynamic and productive. Years to come will likely witness significant changes in perspective and prognosis of affected patients. PMID- 19811957 TI - [Progressive familial intrahepatic cholestasis presenting as liver failure]. AB - Progressive familial intrahepatic cholestasis (PFIC) is a heterogeneous group of autosomic-recessive inherited cholestatic disorders that begin in the neonatal period or in the first years of life. There are three types of PFIC defined by different mutations located in the gene responsible for the bile flow through the intrahepatic canalicular transporter system. These disorders usually present in children or young adults and the main clinical manifestations are cholestasis, jaundice and pruritus, and they progress slowly towards liver fibrosis in adult life. PFIC diagnosis is based on clinical suspicion, biochemical findings (that include normal gamma-glutamyl transpeptidase in type 1 and 2, but increased levels in type 3), image techniques that rule-out other disorders, and histological confirmation. Initial treatment consists of symptomatic relief of cholestatic symptoms with choleretic agents (urso-deoxycholic acid). Partial biliary derivation and ileal bypass are intermediate therapeutic options. In case of no response to these treatments, liver transplantation is indicated. We report the case of a neonate with PFIC type 2 presenting as a liver failure. PMID- 19811958 TI - [Usefulness of a rapid serum interleukin-6 test combined with C-reactive protein to predict sepsis in newborns with suspicion of infection]. AB - AIM: To evaluate the diagnostic value of interleukin-6 (IL-6) to predict the likelihood of neonatal sepsis in order to design an algorithm to decide antibiotic therapy. METHODS: IL-6 and C-reactive protein (CRP) were determined in 42 newborns with clinical suspicion of infection. Newborns were classified as a confirmed, probable or no infection, based on the results of cultures, chest X rays and the involvement of four or more clinical areas on a scale of eight. Samples for IL-6 were collected in the initial assessment and frozen until its determination at the end of the study. Blinded IL-6 measurements were performed using a rapid test. Receiver operator characteristics curves (ROC) for CRP and IL 6 versus infection (confirmed or probable) were determined. RESULTS: Among the 42 cases included in the study 11 (26.2%) were classified as confirmed or probable infection. The area under curve (AUC) for IL-6 was 0.9, with a cut-off value of 53 pg/ml: sensitivity 90.91%, specificity 80%, positive predictive value (PPV) 62.5% and negative (NPV) 96% The level of IL-6>96 pg/ml and/or the combination of IL-6>53+CRP>13.3 mg/l, were the markers that best predicted infection: specificity 100% and PPV: 100%. CONCLUSIONS: Assessment of IL-6 could allow withholding or early discontinuation of antibiotics in newborns with IL-6<54 pg/ml. In cases with IL-6>96 pg/ml and/or IL-6>53+ CRP>13.3, antibiotics should be started promptly, given the high likelihood of infection. Implementation of an algorithm based on the determination of IL-6 and CRP, in the initial assessment of the newborn with clinical suspicion of infection, could reduce unnecessary antibiotic therapy. PMID- 19811959 TI - [Domestic poisoning due to organophosphates]. PMID- 19811960 TI - [Retinopathy of prematurity. Prevention, screening and treatment guidelines]. AB - ROP screening is carried out at all the Spanish hospitals, but there is little agreement on the criteria used for its screening and other aspects related to its practice. Our aim is to report on recommendations for prevention, screening, treatment and follow-up of the retinopathy of prematurity in Spain. Prevention strategies and recommendations for screening, exploration and treatment of ROP, as well as of the organisation of the services to carry out it are reviewed. The most recent bibliography on the basis of the scientific evidence is considered, taking as reference, the guidelines published by the American Academy of Pediatrics (AAP) in 2006 and the recommendations updated in 2007 in the United Kingdom. The recommended prevention strategies are based on the best neonatal practice. The standards, as far as the screening is concerned, are established by birth weight (< or =1,500 g), gestational age (< or =32 weeks) and unstable neonatal clinical course. Indirect ophthalmoscopy is the standard technique for exploration and laser therapy the treatment of choice. PMID- 19811961 TI - [Mycoplasma pneumoniae-induced severe mucositis]. PMID- 19811962 TI - [Loose anagen hair syndrome]. PMID- 19811963 TI - [Predominance of G9 rotavirus in Valencia and Castellon between 2005 and 2007]. AB - INTRODUCTION: Rotavirus is the leading cause of acute gastroenteritis in young children worldwide. Effective vaccines to prevent rotavirus infections are currently available, although their clinical use is still limited, and rotavirus still causes many episodes of infantile gastroenteritis, mainly during the winter seasons. OBJECTIVE: To characterise G (VP7) and P (VP4) genotypes of rotaviruses causing acute gastroenteritis in children and to determine the prevalence of genotype G9 rotavirus in three public health areas in the provinces of Valencia and Castellon. PATIENTS AND METHODS: Five-hundred and forty-one stool samples were prospectively collected from infants and children with gastroenteritis in the period between October 2005 and September 2008. They were analysed for rotavirus by ELISA or by immunochromatography. G and P genotyping was performed by reverse transcription and PCR (RT-PCR). RESULTS: G and P rotavirus genotypes were characterised in a total of 525 faecal samples (97%), resulting in a global predominance of strains G9P[8] (56.5%) and G1P[8] (29.9%). During the period of time studied, G9P[8] was the G/P combination most frequently detected during the rotavirus seasons 2005-2006 and 2006-2007, being present in 81.2% and 64.7% of the patients, respectively. However, during the 2007-2008 season, G1P[8] strains were the most frequently found (68.8%), with a sharp decrease in G9P[8] strains to 7.2% of the samples. CONCLUSIONS: Rotavirus G9P[8] have spread rapidly and widely during the 2005-2006 and 2006-2007 seasons, replacing other previously dominant genotypes (G1, G4) in our geographic area. Its incidence has declined sharply in 2007-2008, in which G1P[8] was again the predominating genotype. PMID- 19811964 TI - Achilles tendon graft for static facial sling. PMID- 19811965 TI - Osteochondral transfer using a transmalleolar approach for arthroscopic management of talus posteromedial lesions. AB - Characterizing osteochondral lesions of the talus has enabled the strategies of surgical management to be better specified. The main technical problem is one of access for arthroscopy instruments to posteromedial lesions. A range of techniques and approaches has been described in ankle arthroscopy in general, and a transmalleolar approach provides reliable and efficient access in these cases. It is frequently used for transchondral drilling, but also enables satisfactory implant positioning in autologous osteochondral mosaicplasty procedures. We report our technique and results on five cases with a minimum 1.2 years' follow up. PMID- 19811966 TI - [From safety to prevention: A(H1N1) influenza, a new challenge for intensivists]. PMID- 19811967 TI - [Can bed sores be avoided in intensive care units?]. PMID- 19811968 TI - [Respiratory work and pattern with different proportional assist ventilation levels]. AB - OBJECTIVE: To study the minimum assistance level during proportional assist ventilation (PAV) to decrease the work of breathing to physiological limits (0.6 j/l) and the relationship between breathing pattern changes and respiratory effort at different PAV levels. DESIGN: Prospective cohort study. SETTING: Polyvalent intensive care unit of a teaching hospital of Jaen, Spain. PATIENTS AND METHODS: Twelve long-term mechanical ventilated patients who met criteria to initiate weaning from the ventilator. INTERVENTIONS: We used the Puritan-Bennett 840 ventilator in proportional assist ventilation. The percentage of support was randomly modified between 5% and 80%, in intervals of 10%. Prior to the change in the PAV level, the patients were ventilated in assist-volume control followed by pressure support ventilation. MAIN VARIABLES OF INTEREST: Before PAV, we measured the respiratory mechanics and the breathing pattern and work of breathing during this mode. RESULTS: The decrease in respiratory assist in PAV was related to significantly higher work of breathing, this going from 0.2+/-0,07 (0.1-0.3) j/l with PAV80 to 0.9+/-0.2 (0.4-1.5) j/l with PAV5 (p=0.002). The coefficient correlation between PAV level and work of breathing (measured as j/l and j/min) was r=-0.8 and -0.6, respectively. Minimum PAV level related with physiological work of breathing was 30% (0.63+/-0.13 j/l). Except for the tidal volume that increased significantly (PAV80 vs PAV5=0.4+/-0.1 vs 0.3+/-0.1; p=0.02), the remaining variables defining the breathing pattern did not changed with the increase in PAV. CONCLUSIONS: In the group of patients studied, the increase in the PAV levels decreases work of breathing, without significantly changing the breathing pattern. Levels lower than 30% of PAV are associated to excessive work of breathing. PMID- 19811969 TI - [Incidence and risk factors associated with the presence of pressure ulcers in critically ill patients]. AB - OBJECTIVE: The incidence of pressure ulcers in the literature varies from 4.7% to 18.6%. In our study, we have aimed to identify the incidence and risk factors associated with the occurrence of pressure ulcers in critically ill patients. SCOPE: Medical-surgical intensive care unit. DESIGN: Prospective observational study in the period of September 2005 to August 2006. PATIENTS AND METHODS: We included patients without PU on admission who were hospitalized more than 48 hours in the intensive care unit (ICU) and who had any of the following risk factors for pressure ulcers: intubated and on mechanical ventilation, with vasopressor support. Main variable interest: The main variable was the presence of grade II or higher PU. Other variables studied were age, weight, body mass index, admission diagnosis, presence of infection, APACHE II (Acute Physiologic and Chronic Health Evaluation), ventilation mechanical level of sedation, presence of diarrhea, wound drainage, vasopressor support, duration of mechanical ventilation, ICU and hospital stay. RESULTS: The study included 150 patients, with an average age of 54 years, the APACHE II averaged 18.2+/-8.2, 88.67% of the patients were on mechanical ventilation, days of stay in ICU were an average of 11.94+/-16.45. Percentage of PU grade II or higher was 26.7%. After the univariate analysis, the risk factors found were presence of infection (OR=2.52 with a p-value=0.013 and a 95% CI, 1.2-5.29), days of stay in ICU (p=0.001; OR=4.39; 95% CI, 6.92-18.25). Presence of infection (OR=2.89; p=0.023; 95% CI, 1.16-7.22), days of stay in the ICU (OR=1.13; p=0.005; 95% CI, 1.06,1.22) and a high score APACHE II (OR=1.06; p=0.044; 95% CI, 1-1.12) were also significant in the multivariate analysis. CONCLUSIONS: The incidence of pressure ulcers in the study population is very high. The risk factors most often found are presence of infection, days of stay and high APACHE II score. PMID- 19811970 TI - [Organ preservation]. AB - Maintaining organ viability from extraction to transplantation is crucial to ensure the function and survival of the graft. In recent years, maintaining organ viability has become more challenging because the shortage of donors has led to broader criteria for donor acceptability and consequently to organs with greater compromise. Organ damage occurs primarily as a result of ischemia-reperfusion injury, which is associated to additional damage from the preservation process. To minimize this damage, different techniques of organ preservation are used with the aim of optimizing organ function once perfusion is restored. Static cold storage is the most commonly used method of preservation because it is extremely simple, nearly universally available, and easy to transport. However, static cold storage may be unable to prevent the deterioration of the quality of organs from donors included under the broader criteria. In this article, we describe current preservation techniques; we place special emphasis on continuous machine perfusion. PMID- 19811971 TI - [Utility of the use of steroids in acute pulmonary dysfunction]. AB - The utility of steroids is a frequent topic of debate in any disease in which an inflammatory component is associated. The physiological rationale for the use of antiinflammatory agents has an important base that is supported by experimental studies in both animals and human beings. However, up to now, its efficiency has not been made clear in the clinical practice. In this article, we aim to summarize and analyze the different studies that evaluate the effect of steroids in ARDS -with different doses and for different moments of the course- that may help to clarify the indication or not of these medications. PMID- 19811972 TI - [A new clinical trial with selective digestive decontamination]. PMID- 19811973 TI - [Review of the post-intubation tracheal stenosis: a case report]. AB - Postintubation tracheal stenosis is a very important clinical situation. It is estimated to occur in approximately 5% to 20% of intubated or tracheostomized patients. However, 1% also suffers severe respiratory dysfunction. We report the case of a 45-year old patient who required surgery under general anesthesia (first intubation experience) after suffering severe coronary disease. The patient was admitted to the intensive care unit, connected to maintained mechanical ventilation for 9 days when he was weaned and extubated. He then suffered a picture of respiratory failure requiring reintubation (for a second time) and reconnection to the mechanical ventilator. He evolved favorably, and it was possible to wean him again with final extubation on the 12th day. At 28 days, he presented a picture of dyspnea, stridor, tachypnea and a bronchoscope examination show tracheal stenosis in the subglotic region. He was treated with laser and silicone tracheal stent, with good evolution. PMID- 19811974 TI - [Acute respiratory distress syndrome secondary to malignant Mediterranean boutonneuse fever]. PMID- 19811975 TI - [Dead space of the central venous catheters as source of errors that may affect the safety of patients in the intensive care unit]. PMID- 19811976 TI - [Considerations on hypnosedation in the postoperative period of pediatric heart surgery]. PMID- 19811977 TI - Influence of composition and pressure on the electric field-induced antiferroelectric to ferroelectric phase transformation in lanthanum modified lead zirconate titanate ceramics. AB - The electric field-induced phase transformation behavior in lanthanum-doped lead zirconate titanate ceramics was examined by polarization versus electrical field (P-E) measurements carried out from room temperature to 130 degrees C and under hydrostatic pressures from 20 to 300 MPa. The samples with composition (Pb(1 x)La(x))(Zr(0.90)Ti(0.10))(1-x/4)O(3) [PLZT x/90/10; x = 2,3,4 at%] were prepared by the standard solid-state reaction method. The analysis at room temperature under atmospheric pressure showed that the increase in the lanthanum content induces a transformation from the typical ferroelectric hysteresis, observed for PLZT 2,3/90/10, to double-hysteresis loops, typical of antiferroelectric phases, for PLZT 4/90/10 under a strong electric field. Hydrostatic pressure- induced and temperature-induced ferroelectric (FE) to antiferroelectric (AFE) phase transformations were examined. The measured hysteresis loops indicated that the FE-AFE phase transformation depends on both temperature and hydrostatic pressure for PLZT 3/90/10. This composition, which is in a ferroelectric state at room temperature under atmospheric pressure, can be transformed into the antiferroelectric phase by the application of a hydrostatic pressure of 100 MPa or by increasing the temperature to around 90 degrees C. The PLZT 2/90/10 and 4/90/10 compositions displayed predominantly ferroelectric and antiferroelectric behavior, respectively, over the ranges of temperature and hydrostatic pressure examined in the present study. PMID- 19811978 TI - Effect of nickel-niobium co-doping on structural, electromechanical, and dielectric properties of lead titanate ceramics. AB - Lead titanate (PT) ceramics co-doped with 8 to 13 mol% Ni-Nb, by substituting titanium ions, were prepared by conventional ceramic method and the influence of Ni-Nb doping level on their morpho-structural, electromechanical and dielectric properties was investigated. Dense tetragonal PT ceramics were obtained for 8 to 12 mol% Ni-Nb co-doping. Lattice anisotropy decreased from 1.053 to 1.046 with increasing doping level. Large thickness coupling factors of 0.4 were obtained for 8, 11, and 12 mol% Ni-Nb. Large electromechanical anisotropy of 10 was obtained for 8 mol% Ni-Nb. It was found that Ni-Nb co-doping has both softening and hardening effects as a function of donor or acceptor behavior of different amounts of Ni-Nb. For 13 mol% Ni-Nb, segregation of a small amount of a secondary phase occurs, lattice constants and tetragonality significantly decrease and the resulting ceramic becomes very hard. PMID- 19811979 TI - Phase transitions in PbZr(x)Hf(1-x)O3 determined by thermal analysis and impedance spectroscopy. AB - The objective of this study is to determine the influence of partial substitutions of Zr by Hf in the perovskitetype crystalline structure of PbZrO(3). Different samples over the whole composition range (0 < or = x < or = 1) in the PbZr(x)Hf(1-x)O(3) family have been prepared. Phase transitions have been determined by thermal analysis (DSC) and complex impedance (IS) spectroscopy over a wide temperature range. As a consequence of the cation replacement, the changes that take place in the different phase transitions temperatures are reported. By both techniques, thermal analysis and electrical characterization, it is shown that for all compositions prepared there are 2 phase transitions in a temperature range between 160 and 230 degrees C. With these results and the previously known crystalline structure of pure PbZrO(3) and PbHfO(3) perovskites, the phase diagram of the PbZr(x)Hf(1-x) O(3) family is presented for the first time. PMID- 19811980 TI - Poled composites with Nd-Mn-doped PZT fibers under bipolar electrical load. AB - Using the sol-gel process, Nd-Mn-doped PZT fibers were produced. The PZT was doped with 2 mol% neodymium and 1.1 mol% manganese. For characterization, the fibers were embedded in a polymer. The resulting 1-3 composites were poled with constant electric field. Strain and polarization were measured by applying a bipolar sinusoidal voltage of high amplitude. Instead of the expected shifted butterfly-shaped strain hysteresis, an asymmetric strain-field relation was observed. It is characterized by a rather linear region in direction of the poling field and an inflated region without strain switching for reversed polarity. Within the temperature range from room temperature to 80 degrees C, the strain switching seems to be suppressed. Measurements of the piezoelectric coefficient at superimposed electric field prove the blocking of strain switching. Cyclation experiments with sesquipolar load show a pronounced linearity of the strain loops that declines after more than 2 x 10(4) cycles. PMID- 19811981 TI - Large-scale fabrication of titanium-rich perovskite PZT submicro/nano wires and their electromechanical properties. AB - We report a 2-step approach to prepare tetragonal perovskite PbZr(0.1)Ti(0.9)O(3) submicro/nano wires in gram scale and with over 95% wire content. Non-perovskite precursor wires were first fabricated by hydrothermal processing. A subsequent annealing in a PbO atmosphere at 600 degrees C converted these wires into perovskite structures which retain the one-dimensional shape. Binding of the perovksite nanowires to a conductive substrate could be achieved by a similar heat treatment of the non-perovskite precursor wires on a flat Pt coated substrate. Taking advantage of the strong mechanical attachment and good electrical contact between the wires and the metallic layer, piezoresponse force microscopy (PFM) was used to measure the local piezoelectric and ferroelectric properties of the individual wires. Enhanced piezoelectric response relative to sputtered epitaxial PbZr(0.2)Ti(0.8)O(3) film and squared hysterisis loop with sharp switching indicate pronounced electro-mechanical and ferroelectric behavior. The 90 degree domain structure of the as-prepared perovskite PZT wires was confirmed by both PFM and transmission electron microscopy investigations. PMID- 19811982 TI - A high temperature piezoelectric ceramic: (1-x)(Bi0.9La0.1)FeO(3-x)PbTiO3 crystalline solutions. AB - (1-x)(Bi(0.9)La(0.1))FeO(3-x)PbTiO(3) (BLF-PT) crystalline solutions for x = 0.35, 0.37, 0.4, 0.43 and 0.45 have been prepared by the solid-state reaction method. The X-ray diffraction analysis shows that BLF-PT has a single perovskite phase with mixed tetragonal and rhombohedral phases between x = 0.4 and 0.43. The Curie temperature of BLF-PT for x = 0.4 attains 460 degrees C, which is about 80 degrees C higher than that of hard Pb(Zr,Ti)O(3) ceramics. The remnant polarization and piezoelectric constant of BLF-PT for x = 0.4 reach 38 microC/cm(2) and 112 pC/N, respectively. The planar coupling factor k(p) of BLF PT for x = 0.4 remains stable at temperature increases of up to 360 degrees C. The impedance spectroscopy study reveals that the high temperature conduction of BLF-PT may be attributed to the motion of oxygen vacancies within the material. Our results indicate that BLF-PT is a promising candidate for high temperature applications. PMID- 19811983 TI - Effects of gallium on the structure and electrical properties of 0.65 (Bi0.94La0.06) (Ga(x)Fe(1-x))O3-0.35PbTiO3 ceramics. AB - Crystalline solutions of 0.65 Bi(0.94)La(0.06)) (Ga(x)Fe(1-x))O(3)-0.35PbTiO(3) Ceramics (BLGF-PT) for x = 0 and 0.05 have been fabricated by the solid-state reaction method. X-ray diffraction (XRD) was utilized to characterize the crystal structure and examine any possible impurities existing in the ceramics. The effects of Ga substitution on dielectric properties of the samples were studied at frequencies from 10(2) to 10(6) Hz over a temperature range from 20 to 620 degrees C. The results indicate that Ga modification can reduce the room temperature dielectric loss. The conduction mechanism of the material was investigated using ac conductivity. It is concluded that electrons originating from Fe(2+) and oxygen ion vacancies are the main charge carriers, and Ga doping could decrease the electronic conduction effectively. The frequency dependence of ac conductivity was found to follow Jonscher's universal power law. PMID- 19811984 TI - Dielectric investigations of 0.945(Bi0.5Na0.5) TiO3-0.055BaTiO3. AB - Dielectric properties of 0.945(Bi(0.5)Na(0.5)) TiO(3)-0.055BaTiO(3)(BNBT5.5) (a composition close to the rhombohedral- tetragonal morphotropic phase boundary) ceramics are studied. It is shown that BNBT5.5 is a relaxor with a characteristic relaxation time that follows Vogel-Fulcher's law. The following Vogel-Fulcher parameters of the relaxation time were calculated: tau(0) = (2.0 +/- 2.4) x 10( 14) s, E/B(k) = (1620 +/- 270) K, Tau(0) = (262 +/- 9) K. PMID- 19811985 TI - Piezoceramics properties as a function of the structure in the system (K,Na,Li)(Nb,Ta,Sb)O3. AB - The influence of the sintering conditions and stoichiometry on the crystalline symmetry and electrical properties of the system (K(0.44+x)Na(0.52)Li(0.04))(Nb(0.86)Ta(0.10)Sb(0.04)) O(3+x/2), with x = -0.06, 0.00, and 0.04, has been evaluated. By lowering the concentration of K cations, a faster stabilization of the tetragonal phase is reached. Increasing the sintering time also leads to the stabilization of the tetragonal phase at room temperature, as evidenced by Raman spectroscopy, thus improving the piezoelectric properties of these materials. The Raman spectra versus temperature showed 2 anomalies at temperatures of approximately 70 and approximately 270 to 330 degrees C, associated with polymorphism and the ferro-paraelectric phase transitions, respectively. Active Raman scattering modes are observed above Curie temperature, indicating that the symmetry is not cubic but pseudocubic. This pseudocubic phase is associated with relaxor diffuseness, attributed to composition fluctuations. In addition, the piezoelectric properties were correlated with the tetragonality of the system, showing a linear dependence between the piezoelectric properties and the tetragonality ratio. The ceramics with c/a = 1.011 ratio exhibit enhanced electrical properties, d(33) approximately 255 pC/N and k(p) approximately 0.47. PMID- 19811986 TI - Lattice dynamics and phase transitions in KNbO3 and K0.5Na0.5NbO3 ceramics. AB - Phonons in both ceramic samples KNbO(3) (KN) and K(0.5)Na(0.5)NbO(3) (KNN-50) were investigated from 10 to 900 K by means of Raman, infrared, and THz spectroscopy. First-order transitions from cubic to tetragonal phases were detected at about 755 K (in KN) and 710 K (in KNN-50), where the first component of the polarization appears. Transitions from the tetragonal to the orthorhombic phases take place around 510 and 475 K, respectively. The last transitions from orthorhombic to rhombohedral phases are strongly first-order type. T(C) is shifted from 200 K in KN to about 90 K in KNN-50. All Raman active modes below 200 cm(-1). disappear in KN but not in KNN-50. The overdamped soft mode present at high temperatures in the THz range changes its dielectric strength at each phase transition (when the corresponding component jumps to higher frequencies) and abruptly disappears from THz spectra in the rhombohedral phase, because it stiffens up to 200 cm(-1). This mode has lower frequency and higher dielectric strength in KNN-50. PMID- 19811987 TI - Radiation of X-rays using polarized LiNbO3 single crystal in low-pressure ambient gas. AB - The dependence of X-ray intensity on the pressure and type of ambient gas was investigated for LiNbO(3) single crystals polarized in the c-axis direction at pressures of approximately 1 to 30 Pa. Ionization of surrounding gas molecules by the electric field generated by the crystal led to the production of both positive ions and free electrons. The electrons were accelerated toward a Cu target, radiating both white X-rays and X-rays specific to the crystal or target material by bremsstrahlung. The integrated X-ray intensity per cycle in the energy range 1 to 20 keV showed a local maximum value at a pressure P(max). The logarithm of P(max) was proportional to the Boltzmann factor using the first ionization energy of each ambient gas molecule. The value of P(max) was found to be independent of the electrical surface area of the crystal. The integrated X ray intensity was approximated qualitatively by a quadratic function with pressure, which was upwardly convex. It was found that one of the causes of the reduction in X-ray intensity at pressures P > P(max) is the adsorption of positive ions generated by the ionization of gas molecules on the negative electric surface. It was also discovered that the lifetime of the X-ray radiation device could be improved when the X-ray radiation case was covered with another hermetically sealed decompression case. The gas with the smallest first ionization energy, with a partial pressure of P(max), was enclosed inside the X ray radiation case (inner case) and the gas with the largest first ionization energy was enclosed at a suitable pressure between the inner and outer cases. PMID- 19811988 TI - Graphics processing unit-based high-frame-rate color Doppler ultrasound processing. AB - Color Doppler ultrasound is a routinely used diagnostic tool for assessing blood flow information in real time. The required signal processing is computationally intensive, involving autocorrelation, linear filtering, median filtering, and thresholding. Because of the large amount of data and high computational requirement, color Doppler signal processing has been mainly implemented on custom-designed hardware, with software-based implementation--particularly on a general-purpose CPU--not being successful. In this paper, we describe the use of a graphics processing unit for implementing signal-processing algorithms for color Doppler ultrasound that achieves a frame rate of 160 fps for frames comprising 500 scan lines x 128 range samples, with each scan line being obtained from an ensemble size of 8 with an 8-tap FIR clutter filter. PMID- 19811989 TI - Capacitance of thin-film ferroelectrics under different drive signals. AB - Thin-film ferroelectric capacitance can be obtained by 2 different methods. Capacitance obtained using the derivative of its hysteresis loop is related to large applied signals and can be called the large-signal capacitance. Capacitance measured directly with a small, applied ac signal together with a slow changing dc bias is called the small-signal capacitance. This paper investigated the voltage dependence of the large- and small-signal capacitances. Measurements show that the large-signal C-V curve of thin-film ferroelectrics has much sharper peaks and higher peak values than the small-signal C-V curve. Analyses based on the Landau-Khalatnikov model shows that practical small-signal capacitance is closer to the ideal capacitance. However, its C-V curve has clearance areas around the coercive voltage, and the polarization switching is not reflected in the small-signal capacitance. This causes the peaks of small-signal C-V curves to be lower than that of large-signal C-V curves. PMID- 19811990 TI - Benefits of minimum-variance beamforming in medical ultrasound imaging. AB - Recently, significant improvement in image resolution has been demonstrated by applying adaptive beamforming to medical ultrasound imaging. In this paper, we have used the minimum-variance beamformer to show how the low sidelobe levels and narrow beamwidth of adaptive methods can be used, not only to increase resolution, but also to enhance imaging in several ways. By using a minimum variance beamformer instead of delay-and-sum on reception, reduced aperture, higher frame rates, or increased depth of penetration can be achieved without sacrificing image quality. We demonstrate comparable resolution on images of wire targets and a cyst phantom obtained with a 96-element, 18.5-mm transducer using delay-and-sum, and a 48-element, 9.25-mm transducer using minimum variance. To increase frame rate, fewer and wider transmit beams in combination with several parallel receive beams may be used. We show comparable resolution to delay-and sum using minimum variance, 1/4th of the number of transmit beams and 4 parallel receive beams, potentially increasing the frame rate by 4. Finally, we show that by lowering the frequency of the transmitted beam and beamforming the received data with the minimum variance beamformer, increased depth of penetration is achieved without sacrificing lateral resolution. PMID- 19811991 TI - Precise time-of-flight calculation for 3-D synthetic aperture focusing. AB - Conventional linear arrays can be used for 3-D ultrasound imaging by moving the array in the elevation direction and stacking the planes in a volume. The point spread function is larger in the elevation plane, because the aperture is smaller and has a fixed elevation focus. Resolution improvements in elevation can be achieved by applying synthetic aperture focusing to the beamformed-in-plane RF data. The proposed method uses a virtual source placed at the elevation focus for postbeamforming. This has previously been done in 2 steps, in-plane focusing followed by synthetic aperture postfocusing in elevation, due to lack of a simple expression for the exact time of flight. This paper presents a new single step method for calculating the time of flight for a 3-D case using a linear array. The new method is more flexible and is able to beamform a fewer number of points much more efficiently. The method is evaluated using both simulated data and phantom measurements using the RASMUS experimental scanner. Computational cost of the method is higher than the 2-step method for a full volume beamforming, but it allows for a reduction of an order-of-magnitude if 3 planes are used for real time visualization. In addition, the need for a temporary storage of beamformed data is removed. PMID- 19811993 TI - Fast computation of the acoustic field for ultrasound elements. AB - A fast method for computing the acoustic field of ultrasound transducers is presented with application to rectangular elements that are cylindrically focused. No closed-form solutions exist for this case but several numerical techniques have been described in the ultrasound imaging literature. Our motivation is the rapid calculation of imaging kernels for physics-based diagnostic imaging for which current methods are too computationally intensive. Here, the surface integral defining the acoustic field from a baffled piston is converted to a 3-D spatial convolution of the element surface and the Green's function. A 3-D version of the overlap-save method from digital signal processing is employed to obtain a fast computational algorithm based on spatial Fourier transforms. Further efficiency is gained by using a separable approximation to the Green's function through singular value decomposition and increasing the effective sampling rate by polyphase filtering. The tradeoff between accuracy and spatial sampling rate is explored to determine appropriate parameters for a specific transducer. Comparisons with standard tools such as Field II are presented, where nearly 2 orders of magnitude improvement in computation speed is observed for similar accuracy. PMID- 19811992 TI - A post-beamforming 2-D pseudoinverse filter for coarsely sampled ultrasound arrays. AB - Beamforming artifacts due to coarse discretization of imaging apertures represent a significant barrier against the use of array probes in high-frequency applications. Nyquist sampling of array apertures dictates center-to-center spacing of lambda/2 for elimination of grating lobes in the array pattern. However, this requirement is hard to achieve using current transducer technologies, even at the lower end of high-frequency ultrasonic imaging (in the range 25-35 MHz). In this paper, we present a new design approach for 2-D regularized pseudoinverse (PIO) filters suitable for restoring imaging contrast in systems employing coarsely sampled arrays. The approach is based on a discretized 2-D imaging model for linear arrays assuming scattering from a Cartesian grid in the imaging field of view (FOV). We show that the discretized imaging operator can be represented with a block Toeplitz matrix with the blocks themselves being Toeplitz. With sufficiently large grid size in the axial and lateral directions, it is possible to replace this Toeplitz-block block Toeplitz (TBBT) operator with its circulant-block block circulant (CBBC) equivalent. This leads to a computationally efficient implementation of the regularized pseudoinverse filtering approach using the 2-D fast Fourier transform (FFT). The derivation of the filtering equation is shown in detail and the regularization procedure is fully described. Using FIELD, we present simulation data to show the 2-D point-spread functions (PSFs) for imaging systems employing linear arrays with fine and coarse sampling of the imaging aperture. PSFs are also computed for a coarsely sampled array with different levels of regularization to demonstrate the tradeoff between contrast and spatial resolution. These results demonstrate the well-behaved nature of the PSF with the variation in a single regularization parameter. Specifically, the 6 dB axial and lateral dimensions of the PSF increase gradually with increasing value of the regularization parameter. On the other hand, the peak grating lobe level decreases gradually with increasing value of the regularization parameter. The results are supported by image reconstructions from a simulated cyst phantom obtained using finely and coarsely sampled apertures with and without the application of the regularized 2-D PIO. PMID- 19811994 TI - Ultrasound transducer and system for real-time simultaneous therapy and diagnosis for noninvasive surgery of prostate tissue. AB - For noninvasive treatment of prostate tissue using high-intensity focused ultrasound this paper proposes a design of an integrated multifunctional confocal phased array (IMCPA) and a strategy to perform both imaging and therapy simultaneously with this array. IMCPA is composed of triple-row phased arrays: a 6-MHz array in the center row for imaging and two 4-MHz arrays in the outer rows for therapy. Different types of piezoelectric materials and stack configurations may be employed to maximize their respective functionalities, i.e., therapy and imaging. Fabrication complexity of IMCPA may be reduced by assembling already constructed arrays. In IMCPA, reflected therapeutic signals may corrupt the quality of imaging signals received by the center-row array. This problem can be overcome by implementing a coded excitation approach and/or a notch filter when B mode images are formed during therapy. The 13-bit Barker code, which is a binary code with unique autocorrelation properties, is preferred for implementing coded excitation, although other codes may also be used. From both Field II simulation and experimental results, we verified whether these remedial approaches would make it feasible to simultaneously carry out imaging and therapy by IMCPA. The results showed that the 13-bit Barker code with 3 cycles per bit provided acceptable performances. The measured -6 dB and -20 dB range mainlobe widths were 0.52 mm and 0.91 mm, respectively, and a range sidelobe level was measured to be 48 dB regardless of whether a notch filter was used. The 13-bit Barker code with 2 cycles per bit yielded -6 dB and -20 dB range mainlobe widths of 0.39 mm and 0.67 mm. Its range sidelobe level was found to be -40 dB after notch filtering. These results indicate the feasibility of the proposed transducer design and system for real-time imaging during therapy. PMID- 19811995 TI - Minimum variance beamforming combined with adaptive coherence weighting applied to medical ultrasound imaging. AB - Currently, the nonadaptive delay-and-sum (DAS) beamformer is used in medical ultrasound imaging. However, due to its data-independent nature, DAS leads to images with limited resolution and contrast. In this paper, an adaptive minimum variance (MV)-based beamformer that combines the MV and coherence factor (CF) weighting is introduced and adapted to medical ultrasound imaging. MV-adaptive beamformers can improve the image quality in terms of resolution and sidelobes by suppressing off-axis signals, while keeping onaxis ones. In addition, CF weighting can improve contrast and sidelobes by emphasizing the in-phase signals and reducing the out-of-phase ones. Combining MV and CF weighting results in simultaneous improvement of imaging resolution and contrast, outperforming both DAS and MV beamformers. In addition, because of the power of CF in reducing the focusing errors, the proposed method presents satisfactory robustness against sound velocity inhomogeneities, outperforming the regularized MV beamformer. The excellent performance of the proposed beamforming approach is demonstrated by several simulated examples. PMID- 19811996 TI - Ultrasonic imaging algorithms with limited transmission cycles for rapid nondestructive evaluation. AB - Imaging algorithms recently developed in ultrasonic nondestructive testing (NDT) have shown good potential for defect characterization. Many of them are based on the concept of collecting the full matrix of data, obtained by firing each element of an ultrasonic phased array independently, while collecting the data with all elements. Because of the finite sound velocity in the test structure, 2 consecutive firings must be separated by a minimum time interval. Depending on the number of elements in a given array, this may become problematic if data must be collected within a short time, as it is often the case, for example, in an industrial context. An obvious way to decrease the duration of data capture is to use a sparse transmit aperture, in which only a restricted number of elements are used to transmit ultrasonic waves. This paper compares 2 approaches aimed at producing an image on the basis of restricted data: the common source method and the effective aperture technique. The effective aperture technique is based on the far-field approximation, and no similar approach exists for the near-field. This paper investigates the performance of this technique in near-field conditions, where most NDT applications are made. First, these methods are described and their point spread functions are compared with that of the Total Focusing Method (TFM), which consists of focusing the array at every point in the image. Then, a map of efficiency is given for the different algorithms in the near-field. The map can be used to select the most appropriate algorithm. Finally, this map is validated by testing the different algorithms on experimental data. PMID- 19811997 TI - Acoustic wave flow sensor using quartz thickness shear mode resonator. AB - A quartz thickness shear mode (TSM) bulk acoustic wave resonator was used for in situ and real-time detection of liquid flow rate in this study. A special flow chamber made of 2 parallel acrylic plates was designed for flow measurement. The flow chamber has a rectangular flow channel, 2 flow reservoirs for stabilizing the fluid flow, a sensor mounting port for resonator holding, one inlet port, and one outlet port for pipe connection. A 5-MHz TSM quartz resonator was edge-bonded to the sensor mounting port with one side exposed to the flowing liquid and other side exposed to air. The electrical impedance spectra of the quartz resonator at different volumetric flow rate conditions were measured by an impedance analyzer for the extraction of the resonant frequency through a data-fitting method. The fundamental, 3rd, 5th, 7th, and 9th resonant frequency shifts were found to be around 920, 3572, 5947, 8228, and 10,300 Hz for flow rate variation from 0 to 3000 mL/min, which had a corresponding Reynolds number change from 0 to 822. The resonant frequency shifts of different modes are found to be quadratic with flow rate, which is attributed to the nonlinear effect of quartz resonator due to the effective normal pressure imposing on the resonator sensor by the flowing fluid. The results indicate that quartz TSM resonators can be used for flow sensors with characteristics of simplicity, fast response, and good repeatability. PMID- 19811998 TI - Determination of epoxy film parameters in a three-layer metal/adhesive/metal structure. AB - The aim of this work is to propose a method to determine the elastic parameters and the thickness of a thin epoxy film located inside a 3-layer aluminum/epoxy/aluminum structure, based on ultrasonic measurements. This study is conducted at low frequencies, to allow the vibration of the whole structure. First, the direct problem is addressed. The sensitivity of the vibration modes to the parameters of interest is studied to select the most sensitive one for a given parameter to be determined. Second, the identification of the parameters with the selected modes is obtained by a minimization of the characteristic equation. This process is applied to experimental data: the longitudinal and shear wave velocities and the thickness of the epoxy film are obtained within a 3% error range. PMID- 19811999 TI - Attenuation of Lamb waves in the vicinity of a forbidden band in a phononic crystal. AB - When a Lamb wave propagates on a plate engraved by a periodic grating, it may exhibit attenuation. This attenuation is related to a coupling of this incident mode with other propagating modes. As the propagation takes place in a periodic medium, the dispersion curves of the modes are of interest because they exhibit passbands and stopbands related to the geometry of the waveguide. The goal of this work is to quantitatively establish the relation between the value of the attenuation of the propagating waves and the width of the forbidden bands appearing inside the Brillouin zone. This study is performed by using a finite element method (ATILA code). PMID- 19812000 TI - Silicon-based megahertz ultrasonic nozzles for production of monodisperse micrometer-sized droplets. AB - Monodisperse ethanol droplets 2.4 microm and water droplets 4.5 microm in diameter have been produced in ultrasonic atomization using 1.5- and 1.0-MHz microelectromechanical system (MEMS)-based silicon nozzles, respectively. The 1.5 and 1.0-MHz nozzles, each consisting of 3 Fourier horns in resonance, measured 1.20 cm x 0.15 cm x .11 cm and 1.79 cm x 0.21 cm x 0.11 cm, respectively, required electrical drive power as low as 0.25 W and could accommodate flow rates as high as 350 microl/min. As the liquid issues from the nozzle tip that vibrates longitudinally at the nozzle resonance frequency, a liquid film is maintained on the end face of the nozzle tip and standing capillary waves are formed on the free surface of the liquid film when the tip vibration amplitude exceeds a critical value due to Faraday instability. Temporal instability of the standing capillary waves, treated in terms of the unstable solutions (namely, time dependant function with a positive Floquet exponent) to the corresponding Mathieu differential equation, is shown to be the underlying mechanism for atomization and production of such monodisperse droplets. The experimental results of nozzle resonance and atomization frequencies, droplet diameter, and critical vibration amplitude are all in excellent agreement with the predictions of the 3-D finite element simulation and the theory of Faraday instability responsible for atomization. PMID- 19812001 TI - Variational principles for polar piezoelectric media in elastic range. AB - The fundamental equations of polar piezoelectric media in differential form are alternatively established in variational forms with their well-known features. First, a 3-field variational principle with some constraint conditions is deduced for a regular region of media from a general principle of physics. The principle is modified by using an involutory transformation and a 9-field variational principle operating on all the field variables is derived. Next, this principle is extended and a unified variational principle is obtained for the region with a fixed internal surface of discontinuity. The unified variational principle is further generalized for the equations of a laminated polar region. The generalized variational principle with the only constraint of initial conditions yields all the equations of the laminae region, including the interface conditions, as its Euler-Lagrange equations. The variational principles are shown to recover some of earlier variational principles, as special cases. PMID- 19812002 TI - Analysis of multifrequency langevin composite ultrasonic transducers. AB - The multimode coupled vibration of Langevin composite ultrasonic transducers with conical metal mass of large cross-section is analyzed. The coupled resonance and anti-resonance frequency equations are derived and the effective electromechanical coupling coefficient is analyzed. The effect of the geometrical dimensions on the resonance frequency, the anti-resonance frequency, and the effective electromechanical coupling coefficient is studied. It is illustrated that when the radial dimension is large compared with the longitudinal dimension, the vibration of the Langevin transducer becomes a multifrequency multimode coupled vibration. Numerical methods are used to simulate the coupled vibration; the simulated results are in good agreement with those from the analytical results. Some Langevin transducers of large cross-section are designed and manufactured and their resonance frequencies are measured. It can be seen that the resonance frequencies obtained from the coupled resonance frequency equations are in good agreement with the measured results. It is expected that by properly choosing the dimensions, multifrequency Langevin transducers can be designed and used in ultrasonic cleaning, ultrasonic sonochemistry, and other applications. PMID- 19812003 TI - Flexible ultrasonic transducers incorporating piezoelectric fibres. AB - It is possible to produce a high-performance, flexible 1-3 connectivity piezoelectric ceramic composite with conventional methods but the process is difficult and time-consuming. Extensive finite element modeling was used to design a piezocomposite structure which incorporated randomly positioned piezoceramic fibers in a polymer matrix. Simple manufacturing techniques were developed which resulted in the production of large numbers of fully populated fiber composites that offered performance comparable with a conventional 1-3 piezocomposite. A modified process facilitated the production of efficient fiber piezocomposite elements separated by polymer channels which conformed to a highly flexible (13 mm radius of curvature), 2-D matrix array configuration. This arrangement has been termed a Composite Element Composite Array Transducer, or CECAT. These devices were evaluated in terms of their impedance spectra, pulse echo response, and surface displacement characteristics. The random piezoceramic fiber arrangements showed comparable sensitivity and bandwidth to periodic devices while minimizing the parasitic interpillar modes associated with periodic structures. Investigations have indicated that CECAT arrays constructed with 250 microm diameter fibers can be operated at frequencies of up to 3 MHz and transducers incorporating 10 microm diameter fibers can extend the frequency range above 6 MHz. Conversely, improved low-frequency devices can be produced with taller pillars than possible with conventional manufacturing techniques. PMID- 19812004 TI - Power harvesting using PZT ceramics embedded in orthopedic implants. AB - Battery lifetime has been the stumbling block for many power-critical or maintenance-free real-time embedded applications, such as wireless sensors and orthopedic implants. Thus a piezoelectric material that could convert human motion into electrical energy provides a very attractive solution for clinical implants. In this work, we analyze the power generation characteristics of stiff lead zirconate titanate (PZT) ceramics and the equivalent circuit through extensive experiments. Our experimental framework allows us to explore many important design considerations of such a PZT-based power generator. Overall we can achieve a PZT element volume of 0.5 x 0.5 x 1.8 cm, which is considerably smaller than the results reported so far. Finally, we outline the application of our PZT elements in a total knee replacement (TKR) implant. PMID- 19812005 TI - Size optimization of a piezoelectric actuator on a clamped elastic plate. AB - We determine by a theoretical analysis the optimal dimension of a piezoelectric actuator attached to a multilayered elastic plate clamped at both ends. The dimension is optimal in the sense of producing maximal plate center flexural displacement. The first-order theory for laminated piezoelectric plates is used. A theoretical solution is obtained. Numerical results calculated from the solution show that when the actuator length takes a particular value, the center flexural displacement of the elastic plate reaches a maximum. For PZT4 and PZT5A actuators this happens when the actuator length is about 55% of the plate length. Basic vibration characteristics of the structure at the first resonance near the optimal actuator length are also calculated. It is found that the admittance is also maximal when the electrode length is optimal. PMID- 19812006 TI - Inverse problem for the geometry profile determination of waveguides with varying section using adiabatic behavior of guided waves. AB - The aim of this work is to obtain the geometry profile of a waveguide of continuous varying section, by inversing the experimental data. This determination is based on the use of the adiabatic behavior of guided waves. The geometry profiles obtained are in good agreement with the exact ones. PMID- 19812007 TI - Linear unbiased prediction of clock errors. AB - In this paper, we propose a new formula for linear unbiased prediction of the local clock timescales. To predict future errors over all the measurement data, a new gain is derived for the p-step ramp unbiased finite impulse response (FIR) predictor. We then show that this gain gives the best linear unbiased fit suitable for forming the prediction vector. The predictor proposed is consistent with linear regression and best linear unbiased estimator. Applications are given for a crystal clock and the USNO Master Clock. PMID- 19812008 TI - [Is there an interest in para-aortic lymphadenectomy in operable endometrial cancer?]. AB - The indication and extent of lymph node dissection in the surgical management of endometrial cancer remains controversial especially concerning the para-aortic lymph nodes. The therapeutic benefit of the lymph node dissection is criticized mainly for low-risk patients for extra-uterine spread. Surgically staging patients is the best method to predict node involvement and it allows an optimal decision for adjuvant therapy to be taken. The different prognostic factors for para-aortic lymph nodes metastasis are histological grade and size of the tumour, myometrial wall invasion and lymphovascular dissemination, as well as positive pelvic lymph nodes. However, these elements are not correctly evaluated before and during the surgery. Positive para-aortic lymph nodes can be found without a lymphatic spread to the pelvic area. Even though the prevalence of para-aortic node involvement is weak, it seems legitimate to propose in selected cases of important lymph node involvement, it's complete dissection if a pelvic lymphadenectomy is indicated and if it is surgically possible. PMID- 19812009 TI - [Specificity of paediatric oncology pharmacology]. AB - Anticancer chemotherapy plays a key role in the treatment of cancer in children. Following its increased efficacy, three children out of four can now be cured. Nevertheless, given the fact that 25% of the children with cancer are not cured and chemotherapy-induced long-term side effects are numerous there is a need to keep developing new agents and new strategies to fight cancer. Moreover studies investigating off-patents drugs should be stimulated in paediatric oncology in order to improve our knowledge of "old" drugs. Here we will review the specificities of children as compared to adults in the context of clinical drug development and pharmacology research in paediatric oncology. Such a research is now encouraged and facilitated by a European regulation as well as by international consortia such as "Innovative Therapies for Children with Cancer" (ITCC). PMID- 19812010 TI - [The role of real-time three-dimensional echocardiography in the evaluation of hypertrophic cardiomyopathy]. AB - Hypertrophic cardiomyopathy is a relatively common hereditary disorder, which is associated with cardiac morphologic and functional alterations. Echocardiography is a non-invasive, simple and easy-to-learn method to evaluate patients with cardiomyopathy. The aim of this review paper is to demonstrate the possible diagnostic role of one of the newest echocardiographic development, the real-time 3-dimensional echocardiography in the evaluation of hypertrophic cardiomyopathy. PMID- 19812011 TI - [The value of routine second-look endoscopy in the management of acute gastroduodenal ulcer bleeding]. AB - The role of routine second-look endoscopy in the management of patients with acute peptic ulcer bleeding is controversial. A more precise identification of higher risk patient group, based on both clinical and endoscopic criteria, is needed to determine whether there are high-risk patients who may benefit from this management strategy. AIM: Or aim was to find out whether scheduled second look endoscopy has any beneficial effect in the clinical outcome. METHODS: Both endoscopic and clinical data were analyzed in 274 acute gastroduodenal ulcer bleeding patients. The need for repeated endoscopic haemostatic intervention was used as a measure to evaluate the potential beneficial effect of the second look endoscopy. Patients were categorized according to the Forrest classification detected during the emergency endoscopy. RESULTS: In the subgroup of actively bleeding patients (Forrest Ia, Ib) a second endoscopic haemostasis was performed in 23.8% of cases. In the patient subgroup with visible vessel ulcers (Forrest IIa) and in those with adherent clot covered ulcers (Forrest IIb) the needs for a repeated haemostasis were 13.0% and 13.3% respectively. Despite the not statistically significant differences, remarkable clinical impact was noted favoring scheduled second look endoscopy in patients with initially active ulcer bleeding. CONCLUSION: In the light of the retrospective study results it may be concluded that the scheduled second look endoscopy strategy offers a beneficial clinical outcome for selected patients estimated to be a very high risk of re bleeding following the initial endoscopic therapy for active bleeding. PMID- 19812012 TI - [Therapeutic management of central nervous system lymphomas in a single hematological institute]. AB - Primary central nervous system lymphoma is defined as an extranodal lymphoma arising in the central nervous system in the absence of systemic disease. Because of their rare occurrence among lymphomas, optimal treatment could hardly be established. AIMS: In this retrospective survey we analyzed the result of combined treatment (systemic and intrathecal chemotherapy followed by consolidation radiotherapy) in patients with primary or relapsed central nervous system lymphomas diagnosed and treated in our hematological department between 1998-2009. PATIENTS AND METHODS: During this period (mean follow-up of 13.2 months) from 427 patients with newly diagnosed non-Hodgkin's lymphomas, 22 primary central nervous system lymphoma was diagnosed (5.15%, 16 cerebral and 6 spinal cord lymphoma cases). Significant male predominance (14:8) was registered with an age distribution of 34-77 years (mean = 60.7, median = 64 years). No patients were immunocompromised. All central nervous system lymphoma specimens taken with neurosurgical resection or stereotaxic biopsies were confirmed histopathologically. All cerebral lymphoma cases proved to be diffuse large B cell of origin, while in epidural lymphomas low grade subtypes also occurred. Epidural lymphomas were treated with local radiotherapy (30-40 Gy), except for patients with follicular lymphomas getting rituximab-containing polychemotherapy (R + CHOP regimen) before irradiation. In cerebral lymphoma (every patients had supratentorial localization) the following combined therapy protocol was used: up to three courses of high dose methotrexate (HD MTX 3g/m 2 in a single dose for 4 hours lasting drop-infusion) were given at 4-week intervals, followed by leucovorin-rescue 24 hours after MTX infusion. Intrathecal combination of methotrexate, cytosin-arabinosid and dexamethasone was given three times after HD MTX infusion. In complete response after chemotherapy (evaluated by cranial MRI or CT, PET/CT), whole-brain irradiation was used in a total dose of 30 Gy. In case of partial response, boost irradiation for the tumor bed was also given. In relapse or resistant cases, salvage regimen was applied: HD MTX course combined with high dose cytosin-arabinosid (HD Ara-C) 3g/m 2 /dose b.i.d. over 4 h c.i., repeated in three cycles every four weeks. RESULTS: Complete remission has been achieved in 9 patients with cerebral and in 4 patients with spinal cord lymphoma (13/22; 59.0%), however, one relapsed patient became resistant and later expired, despite salvage therapy. Primarily 9 patients were not evaluable for response: 5 received only one or two HD MTX because of side effects, 4 patients died due to progression of the disease. Mean of the overall survival (OS) in cerebral lymphoma was 19.5 (3-46, median of 10) months, in epidural group 14.1 (2-76, median of 5) months, whilst mean time to progression (TTP) was 4.5 (2-6.5, median of 4 months). The 2-year survival for all patients was 50%. Acute toxicity of chemotherapy was usually hematological, moreover, in 8 patients impaired renal function and sepsis developed. No serious adverse effect of radiotherapy could be detected. CONCLUSION: In primary central nervous system lymphoma, basic treatment HD methotrexate together with intrathecal combination of methotrexate + cytosin arabinosid + dexamethasone followed by whole-brain irradiation of at least 30 Gy could produce a medium response rate in our study. In case of relapse or progression, other salvage regimens containing HD Ara-C alternating with HD MTX could reduce the treatment failure, as well. After therapy PET/CT was negative in five patients with prolonged disease-free survival. PMID- 19812013 TI - [Anaphylactic reaction caused by a horse-fly species (Hippobosca equina)]. AB - The authors report on the case of a 46-year-old female patient who had an anaphylactic reaction of lucky outcome after a fly bite. She shortly developed a hard swelling of 15 cm diameter covered by scab, on the hairy scalp where hair meets the forehead, as well as an oedema on the forehead. Shortly afterwards came an erythema and pruritus on the face and body, and oedema on the hands, face and lips, later accompanied by shivers, nausea and vomit. The captured insect was identified by a taxonomic key to be a Hippobosca equina (horse-fly). To our knowledge, this is the first case reported in Hungary on anaphylactic reactions to Hippobosca equina bite. Hippobosca equina lives all over the world - also known in Hungary -, mostly in grazing lands, stables and cowsheds, sucking the blood of horses, asses and occasionally other animals. As a vector it may be a carrier and transmitter of pathogens. With this report we wish to call attention of family physicians and clinicians on the clinical symptoms of the bite of Hippobosca equina, since they have not the specific experience required to the diagnosis of such insect bite. PMID- 19812014 TI - [In memoriam Endre Goth (1909-1976)]. PMID- 19812015 TI - [Karoly Lechner and his Transylvanian intellectual heritage]. PMID- 19812017 TI - [Investigation of adolescents' bone metabolism in the western part of Transdanubia]. AB - Childhood reference range based on the age is not available in Hungary, therefore the diagnosis and therapy of bone metabolic diseases of childhood are subject to difficulties. The aim of this work is to provide information about the adolescents' results of bone mineral density and bone biomarkers. SUBJECTS AND METHODS: Measurements were performed in 169 healthy adolescents (98 girls, 71 boys, age: 17.0+/-1.2 years). Bone mineral content (BMC) and bone mineral density (BMD) of the lumbar spine were measured using Double X-ray Absorptiometry (DXA, LUNAR, GE Health Care, USA) and Z-score values were analyzed using different reference population. In the serum, bone biomarkers osteocalcin (OC) and beta crosslaps (beta-Cl) were measured by a fully automated, electrochemiluminescence immunoassay method (Elecsys-2010, Roche). Data were analyzed according to gender and the Tanner stage and grade system. Associations between body mass index (BMI), calcium intake, consumption of soft drinks and coke, and physical exercise were investigated. RESULTS: BMC values for both age groups were significantly elevated in boys of the Tanner stage V. (15-16 years: 62.9+/-14.3 g; 17-19 years: 69.8+/-9.3g) than in girls (58.1+/-10.4; 61.6+/-8.5 g) (p<0.001). BMD values were higher in girls, than in boys (1.17+/-0.12 g/cm 2 vs. 1.13+/-0.11 g/cm 2) (p<0.05). OC and beta-Cl levels showed negative correlation with age in both gender (p<0.01), while OC and beta-Cl levels were higher in boys, than in girls (p<0.001). Elevation of BMC and BMD values were associated with increase of BMI in both gender (p<0.05), but the biomarkers in thin girls were higher, than in overweight girls (p<0.05). Authors obtained excellent correlations between the BMD-Z-score values compared to the German standard and to their own population (girls: r=0.97, boys: 0.88), but the absolute values significantly differed from one another. 80% of adolescents are on a diet with insufficient calcium intake, while 38% of them do not play sport regularly. Excessive intake of soft drinks was determined in 60% of adolescents. In the case of insufficient calcium intake (4.7%, 6/127), low bone mass was measured using the Z-score of the German reference values. Among children with adequate calcium intake, BMD assessed by DXA was normal. CONCLUSION: These data help to determine normal reference values among healthy high school students. Further studies are needed in wider range of young population for the establishment of Hungarian reference values of bone markers. PMID- 19812018 TI - [Mortality of patients admitted to hospital with acute ST-elevation myocardial infarction, before and after the opening of primary percutaneous coronary intervention unit in Szombathely, Hungary]. AB - Treatment of acute myocardial infarction has changed in recent years. Tremendous debate has developed over the efficacy of percutaneous coronary intervention compared with fibrinolysis. If primary percutaneous coronary intervention is available, it is the preferred treatment for acute ST segment elevation myocardial infarction. AIM: Three months mortality of patients admitted to hospital with ST elevation myocardial infarction in city of Szombathely was analyzed. METHODS: Mortality rates of two time periods were compared: year 2005 without primary percutaneous coronary intervention capability and year 2008 when local primary percutaneous coronary intervention was available. RESULTS: In patient group with no longer than 12 hours ischaemic period 3 months mortality rate was lower in 2008 compared to 2005 (3.6% versus 15.6%). First of all, the relative high 3 months mortality rate in patient group treated with fibrinolytic therapy was responsible for this difference. Decreasing number of patients with longer than 12 hours ischaemic time in 2008 versus 2005 was also important. CONCLUSION: Implementation of primary percutaneous coronary intervention for the management of ST segment elevation myocardial infarction patients in Szombathely was effective for patients' outcome. PMID- 19812019 TI - [The value of twenty-four hour intraoesophageal pH monitoring and manometry in the management of patients with upper gastrointestinal symptoms]. AB - Functional gastroenterological examinations (intraoesophageal pH monitoring, oesophageal manometry, scintigraphy, impedance examination) play important role in the management of patients with upper gastrointestinal complaints. PATIENTS AND METHODS: Four different cases are demonstrated where diagnose and therapy was developed by these examinations. Two patients had typical gastro-oesophageal reflux symptoms and two others had dysphagia. Intraoesophageal pH monitoring was performed by Zinetics twenty-four hour one or two channel pH catheters and oesophageal manometry was carried out by Zinetics EMC four channel catheter with water perfusion method. CASE REPORTS: In one of the patients with typical and extraoesophageal reflux symptoms, lower oesophageal sphincter incompetency by manometry and pathological acid reflux was observed by intraoesophageal pH monitoring, respectively. Furthermore, hiatal hernia was established, peristalsis of the oesophagus proved to be preserved. Because of incomplete efficacy of proton pump inhibitor (PPI) therapy, antireflux surgery was indicated. An other patient with reflux symptoms had physiological pH monitoring and manometric values. Hypersensitive oesophagus was diagnosed and PPI therapy in double dose was applied. Both patients are symptom free up to now. Other two patients complained difficult swallowing and weight loss. Absence of lower oesophageal sphincter relaxation and hypomotility of the oesophagus was observed. After oesophageal dilatation, both patients with achalasia could easy swallow and eat. CONCLUSIONS: Our cases confirm the importance of the twenty-four hour intraoesophageal pH monitoring and oesophageal manometry in the diagnosis of gastro-oesophageal reflux disease, non-cardiac chest pain, other extraoesophageal manifestations and dysphagia. These examinations support the decision for the adequate therapeutic strategy (conventional treatment, surgery or operation or endoscopic intervention) and are important in the follow-up of patients. PMID- 19812020 TI - [Primary, definitive treatment of the nasoethmoidorbital-midface-frontobasal injuries in the maxillofacial practice]. AB - Nasoethmoid-orbital (NEO) fractures are typically resulted by a drastic, forceful blow to the central aspect of the head and nose, associated injuries to adjacent structures occur and involve the midface, anterior cranial fossa and the eye. Dislocated bony parts of the nose may be impacted into the intraorbital region and lead to severe complications: cerebrospinal fluid leak, optic nerve lesion, anosmy, telecanthus and diplopy. In the complex management of the craniofacial lesions, a cooperative team-work of maxillofacial surgeon, neurosurgeon, otolaryngologist and ophthalmologist is required. The key of the surgery treatment is the actual status of the NEO structure. Stabilized nasoethmoidal fragments, midface and the watertight dura closure or plasty are basically important conditions for the ceasing of nasal liquorrhoea. Repositioning of fractured fragments, stabilization with titanium miniplates and reconstruction of bony defect using autologous bone are commonly used techniques at our practice. Between years 1995-2000, altogether 50 cases of injuries due to NEO fracture have been treated at our department of maxillofacial surgery. We report the possible combinations of traumatism, our operative methods, our supply tactics and experiences. Multiple and defect fractures of the craniofacial bones may cause irreversible deformities, chewing and nutritional disturbances. We preferred the early, primary, definitive supply, with the respect of aesthetic viewpoints with the application of streamlined operative methods like bone fixation and bone replacement. Our fundamental aim is the reduction of the patients' operative load, their nursing time and the enhancement of our operative results. PMID- 19812021 TI - [Experiences with cardiac troponin assessments]. AB - After the JESC/ACCC guideline (approved in year 2000), cardiac troponins became the first-line laboratory markers in the diagnosis of acute coronary syndrome. Since 2006, there is a hemodynamic laboratory in our Hospital and since July, 2007, there is a so called "one gate" Emergency Unit. AIM: Since the amount of the cardiac troponin assessments has risen, we aimed to analyze the background of this elevation, the results and the cause of the false positivity, as well. METHODS: We analyzed our cardiac troponin results assessed in the second half of year 2008. RESULTS: In this time we have had 2656 cardiac troponin examination in 1787 patients. 280 positive results were examined in 203 patients, but acute coronary syndrome was proved only in 110 patients. The remaining cases were false positive but only for detecting acute coronary syndrome, because many other diseases were proved in the background of these patients, and only one result was really (literally) false positive. CONCLUSIONS: Because of the very low rate of positive results we have to think about restriction of asking that examination. We emphasize that there are many non-acute coronary syndrome diseases with positive cardiac troponin results. This situation needs an urgent and close communication between the clinicians and the laboratory staff. PMID- 19812022 TI - [Trichotillomania]. AB - The impairment and distress due to trichotillomania should not be underestimated. It can be associated with serious sociological and psychological effects (e.g. strong feelings of shame and embarrassment) as well as avoidance behavior including potentially dangerous avoidance of medical care. It can easily lead into a significant decline in quality of life for patients and their family members. In the background, violence or abuse can often be ruled out, but can be associated with other disease such as mental retardation, schizophrenia, autism and drug-abuse. For the effective treatment collaboration with the family, involving the help of a psychologist or a psychiatrist is required. PMID- 19812023 TI - [Two faces of an illness: rheumatoid arthritis followed by ankylosing spondylitis]. AB - Author presents the clinical history of a 35 years old female patient, who suffered from seronegative rheumatoid arthritis at the age of 29. Remission of the illness was followed by successful gravidity. Almost two years later spondylarthritis was diagnosed, showing only axial symptoms. The major histocompatibility complex analyzed by normal serological methods revealed the HLA B27 haplotype, typical for ankylosing spondylitis, and the HLA DR1 haplotype, typical for rheumatoid arthritis. During the examination of HLA DRB polymorphism, presence of DR B1 0101 allele was found, that plays role in the pathophysiology of rheumatoid arthritis. This case proves that a combination of HLA alleles typical for ankylosing spondylitis and rheumatoid arthritis can be found in the same patient. PMID- 19812025 TI - Cognitive coping and goal adjustment after first-time myocardial infarction: relationships with symptoms of depression. AB - The authors studied the relationships among cognitive coping strategies, goal adjustment processes (disengagement and reengagement), and depressive symptomatology in a sample of 139 patients who had experienced a first-time acute myocardial infarction between 3 and 12 months before data assessment. They assessed cognitive coping strategies, goal adjustment, and depressive symptoms by the Cognitive Emotion Regulation Questionnaire, the Goal Obstruction Questionnaire, and the Hospital Anxiety and Depression Scale, respectively. Main statistical methods were Pearson correlations and multiple regression analyses. Results show significant associations among the cognitive coping strategies of rumination, catastrophizing, and higher depressive symptoms, as well as among positive refocusing, goal reengagement, and lower depressive symptoms. This suggests that cognitive coping and goal reengagement strategies may be useful targets for intervention. PMID- 19812026 TI - Stress and coping styles are associated with severe fatigue in medical students. AB - Fatigue is a common complaint among medical students and researchers consider it to be related to poor academic outcomes. The authors' goal in the present study was to determine whether stress and coping strategies were associated with fatigue in medical students. The study group consisted of 73 second-year healthy students attending the Osaka City University Graduate School of Medicine. Participants completed a questionnaire about fatigue (Japanese version of Chalder Fatigue Scale), stress, stress coping (Japanese version of the Coping Inventory for Stressful Situations), overwork, and nocturnal sleeping hours. On univariate and multivariate logistic regression analyses adjusted for age and gender, stress was positively associated with fatigue. In addition, after adjustment for age, gender, and emotion- and task-oriented stress coping activities, avoidance oriented stress coping activity was associated with fatigue. The results suggest that stress and the coping style are correlated with fatigue in medical students. PMID- 19812027 TI - Key factors in smoking cessation intervention among 15-16-year-olds. AB - The authors aimed to investigate factors associated with smoking cessation among adolescents after tobacco intervention. They examined smokers (n = 127) from one birth cohort (n = 545) in the city of Kotka in Finland. These smokers were randomized in 3 intervention groups the dentist (n = 44) and the school nurse (n = 42 groups), and a control group (n = 39). After 2 months, the authors sent a follow-up questionnaire to the initial smokers to find out who had quit.The authors found that those whose best friend was a nonsmoker were more likely to stop smoking (relative risk RR 7.0 95% Cl 4.6-10.7). Moreover, the nicotine dependent participants (measured according to the Fagerstrom Test for Nicotine Dependence(36)) were less likely to stop (RR 0.1 95% Cl 0.08-0.11) compared to non-nicotine dependent participants. Last, of the diurnal types, the morning types found it easier to quit smoking than the evening types (RR 2.2 95% Cl 1.4 3.6). Thus, the authors concluded that the best friend''s influence, nicotine dependence, and diurnal type could be taken more into account in individual counseling on smoking cessation. PMID- 19812028 TI - The Cytophaga hutchinsonii ChTPSP: First characterized bifunctional TPS-TPP protein as putative ancestor of all eukaryotic trehalose biosynthesis proteins. AB - The most widely distributed pathway to synthesize trehalose in nature consists of two consecutive enzymatic reactions with a trehalose-6-P (T6P)-synthase (TPS) enzyme, producing the intermediate T6P, and a T6P-phosphatase (TPP) enzyme, which dephosphorylates T6P to produce trehalose and inorganic phosphate. In plants, these enzymes are called Class I and Class II proteins, respectively, with some Class I proteins being active enzymes. The Class II proteins possess both TPS and TPP consensus regions but appear to have lost enzymatic activity during evolution. Plants also contain an extra group of enzymes of small protein size, of which some members have been characterized as functional TPPs. These Class III proteins have less sequence similarity with the Class I and Class II proteins. Here, we characterize for the first time, by using biochemical analysis and yeast growth complementation assays, the existence of a natural TPS-TPP bifunctional enzyme found in the bacterial species Cytophaga hutchinsonii. Through phylogenetic analysis, we show that prokaryotic genes such as ChTPSP might be the ancestor of the eukaryotic trehalose biosynthesis genes. Second, we show that plants have recruited during evolution, possibly by horizontal transfer from bacteria such as Rhodoferax ferrireducens, a new type of small protein, encoding TPP activity, which have been named Class III proteins. RfTPP has very high TPP activity upon expression in yeast. Finally, we demonstrate that TPS gene duplication, the recruitment of the Class III enzymes, and recruitment of an N terminal regulatory element, which regulates the Class I enzyme activity in higher plants, were initiated very early in eukaryan evolution as the three classes of trehalose biosynthesis genes are already present in the alga Ostreococcus tauri. PMID- 19812029 TI - Footprints of X-to-Y gene conversion in recent human evolution. AB - Different X-homologous regions of the male-specific portion of the human Y chromosome (MSY) are characterized by a different content of putative single nucleotide polymorphisms (SNPs), as reported in public databases. The possible role of X-to-Y nonallelic gene conversion in contributing to these differences remains poorly understood. We explored this issue by analyzing sequence variation in three regions of the MSY characterized by a different degree of X-Y similarity and a different density of putative SNPs: the PCDH11Y gene in the X-transposed (X Y identity 99%, high putative SNP content); the TBL1Y gene in the X-degenerate (X Y identity 86-88%, low putative SNP content); and VCY genes-containing region in the P8 palindrome (X-Y identity 95%, low putative SNP content). Present findings do not provide any evidence for gene conversion in the PCDH11Y and TBL1Y genes; they also strongly suggest that most putative SNPs of the PCDH11Y gene (and possibly the entire X-transposed region) are most likely X-Y paralogous sequence variants, which have been entered in the databases as SNPs. On the other hand, clear evidence for the VCY genes in the P8 palindrome having acted as an acceptor of X-to-Y gene conversion was obtained. A rate of 1.8 x 10(-7) X-to-Y conversions/bp/year was estimated for these genes. These findings indicate that in the VCY region of the MSY, X-to-Y gene conversion can be highly effective to increase the level of diversity among human Y chromosomes and suggest an additional explanation for the ability of the Y chromosome to retard degradation during evolution. Present data are expected to pave the way for future investigations on the role of nonallelic gene conversion in double-strand break repair and the maintenance of Y chromosome integrity. PMID- 19812030 TI - Characterization of a serine protease homologous to house dust mite group 3 allergens from the scabies mite Sarcoptes scabiei. AB - The scabies mite, Sarcoptes scabiei var. hominis, infests human skin, causing allergic reactions and facilitating bacterial infection by Streptococcus sp., with serious consequences such as rheumatic fever and rheumatic heart disease. To identify a possible drug target or vaccine candidate protein, we searched for homologues of the group 3 allergen of house dust mites, which we subsequently identified in a cDNA library. The native protein, designated Sar s 3, was shown to be present in the mite gut and excreted in fecal pellets into mite burrows within the upper epidermis. The substrate specificity of proteolytically active recombinant rSar s 3 was elucidated by screening a bacteriophage library. A preference for substrates containing a RS(G/A) sequence at the P1-P2' positions was revealed. A series of peptides synthesized as internally quenched fluorescent substrates validated the phage display data and high performance liquid chromatography/mass spectrometry analysis of the preferred cleaved substrate and confirmed the predicted cleavage site. Searches of the human proteome using sequence data from the phage display allowed the in silico prediction of putative physiological substrates. Among these were numerous epidermal proteins, with filaggrin being a particularly likely candidate substrate. We showed that recombinant rSar s 3 cleaves human filaggrin in vitro and obtained immunohistological evidence that the filaggrin protein is ingested by the mite. This is the first report elucidating the substrate specificity of Sar s 3 and its potential role in scabies mite biology. PMID- 19812031 TI - Regulation of TRPC1 and TRPC4 cation channels requires an alpha1-syntrophin dependent complex in skeletal mouse myotubes. AB - The dystrophin-associated protein complex (DAPC) is essential for skeletal muscle, and the lack of dystrophin in Duchenne muscular dystrophy results in a reduction of DAPC components such as syntrophins and in fiber necrosis. By anchoring various molecules, the syntrophins may confer a role in cell signaling to the DAPC. Calcium disorders and abnormally elevated cation influx in dystrophic muscle cells have suggested that the DAPC regulates some sarcolemmal cationic channels. We demonstrated previously that mini-dystrophin and alpha1 syntrophin restore normal cation entry in dystrophin-deficient myotubes and that sarcolemmal TRPC1 channels associate with dystrophin and the bound PDZ domain of alpha1-syntrophin. This study shows that small interfering RNA (siRNA) silencing of alpha1-syntrophin dysregulated cation influx in myotubes. Moreover, deletion of the PDZ-containing domain prevented restoration of normal cation entry by alpha1-syntrophin transfection in dystrophin-deficient myotubes. TRPC1 and TRPC4 channels are expressed at the sarcolemma of muscle cells; forced expression or siRNA silencing showed that cation influx regulated by alpha1-syntrophin is supported by TRPC1 and TRPC4. A molecular association was found between TRPC1 and TRPC4 channels and the alpha1-syntrophin-dystrophin complex. TRPC1 and TRPC4 channels may form sarcolemmal channels anchored to the DAPC, and alpha1 syntrophin is necessary to maintain the normal regulation of TRPC-supported cation entry in skeletal muscle. Cation channels with DAPC form a signaling complex that modulates cation entry and may be crucial for normal calcium homeostasis in skeletal muscles. PMID- 19812032 TI - Structural basis for the role of the K65R mutation in HIV-1 reverse transcriptase polymerization, excision antagonism, and tenofovir resistance. AB - K65R is a primary reverse transcriptase (RT) mutation selected in human immunodeficiency virus type 1-infected patients taking antiretroviral regimens containing tenofovir disoproxil fumarate or other nucleoside analog RT drugs. We determined the crystal structures of K65R mutant RT cross-linked to double stranded DNA and in complexes with tenofovir diphosphate (TFV-DP) or dATP. The crystals permit substitution of TFV-DP with dATP at the dNTP-binding site. The guanidinium planes of the arginines K65R and Arg(72) were stacked to form a molecular platform that restricts the conformational adaptability of both of the residues, which explains the negative effects of the K65R mutation on nucleotide incorporation and on excision. Furthermore, the guanidinium planes of K65R and Arg(72) were stacked in two different rotameric conformations in TFV-DP- and dATP bound structures that may help explain how K65R RT discriminates the drug from substrates. These K65R-mediated effects on RT structure and function help us to visualize the complex interaction with other key nucleotide RT drug resistance mutations, such as M184V, L74V, and thymidine analog resistance mutations. PMID- 19812033 TI - Allosteric inhibition of human porphobilinogen synthase. AB - Porphobilinogen synthase (PBGS) catalyzes the first common step in tetrapyrrole (e.g. heme, chlorophyll) biosynthesis. Human PBGS exists as an equilibrium of high activity octamers, low activity hexamers, and alternate dimer configurations that dictate the stoichiometry and architecture of further assembly. It is posited that small molecules can be found that inhibit human PBGS activity by stabilizing the hexamer. Such molecules, if present in the environment, could potentiate disease states associated with reduced PBGS activity, such as lead poisoning and ALAD porphyria, the latter of which is associated with human PBGS variants whose quaternary structure equilibrium is shifted toward the hexamer (Jaffe, E. K., and Stith, L. (2007) Am. J. Hum. Genet. 80, 329-337). Hexamer stabilizing inhibitors of human PBGS were identified using in silico prescreening (docking) of approximately 111,000 structures to a hexamer-specific surface cavity of a human PBGS crystal structure. Seventy-seven compounds were evaluated in vitro; three provided 90-100% conversion of octamer to hexamer in a native PAGE mobility shift assay. Based on chemical purity, two (ML-3A9 and ML-3H2) were subjected to further evaluation of their effect on the quaternary structure equilibrium and enzymatic activity. Naturally occurring ALAD porphyria-associated human PBGS variants are shown to have an increased susceptibility to inhibition by both ML-3A9 and ML-3H2. ML-3H2 is a structural analog of amebicidal drugs, which have porphyria-like side effects. Data support the hypothesis that human PBGS hexamer stabilization may explain these side effects. The current work identifies allosteric ligands of human PBGS and, thus, identifies human PBGS as a medically relevant allosteric enzyme. PMID- 19812034 TI - Cyclin E is stabilized in response to replication fork barriers leading to prolonged S phase arrest. AB - Cyclin E is a regulator of cyclin-dependent protein kinases (Cdks) and is involved in mediating the cell cycle transition from G(1) to S phase. Here, we describe a novel function for cyclin E in the long term maintenance of checkpoint arrest in response to replication barriers. Exposure of cells to mitomycin C or UV irradiation, but not ionizing radiation, induces stabilization of cyclin E. Stabilization of cyclin E reduces the activity of Cdk2-cyclin A, resulting in a slowing of S phase progression and arrest. In addition, cyclin E is shown to be required for stabilization of Cdc6, which is required for activation of Chk1 and the replication checkpoint pathway. Furthermore, the stabilization of cyclin E in response to replication fork barriers depends on ATR, but not Nbs1 or Chk1. These results indicate that in addition to its well studied role in promoting cell cycle progression, cyclin E also has a role in regulating cell cycle arrest in response to DNA damage. PMID- 19812035 TI - A pathogenic C terminus-truncated polycystin-2 mutant enhances receptor-activated Ca2+ entry via association with TRPC3 and TRPC7. AB - Mutations in PKD2 gene result in autosomal dominant polycystic kidney disease (ADPKD). PKD2 encodes polycystin-2 (TRPP2), which is a homologue of transient receptor potential (TRP) cation channel proteins. Here we identify a novel PKD2 mutation that generates a C-terminal tail-truncated TRPP2 mutant 697fsX with a frameshift resulting in an aberrant 17-amino acid addition after glutamic acid residue 697 from a family showing mild ADPKD symptoms. When recombinantly expressed in HEK293 cells, wild-type (WT) TRPP2 localized at the endoplasmic reticulum (ER) membrane significantly enhanced Ca(2+) release from the ER upon muscarinic acetylcholine receptor (mAChR) stimulation. In contrast, 697fsX, which showed a predominant plasma membrane localization characteristic of TRPP2 mutants with C terminus deletion, prominently increased mAChR-activated Ca(2+) influx in cells expressing TRPC3 or TRPC7. Coimmunoprecipitation, pulldown assay, and cross linking experiments revealed a physical association between 697fsX and TRPC3 or TRPC7. 697fsX but not WT TRPP2 elicited a depolarizing shift of reversal potentials and an enhancement of single-channel conductance indicative of altered ion-permeating pore properties of mAChR-activated currents. Importantly, in kidney epithelial LLC-PK1 cells the recombinant 679fsX construct was codistributed with native TRPC3 proteins at the apical membrane area, but the WT construct was distributed in the basolateral membrane and adjacent intracellular areas. Our results suggest that heteromeric cation channels comprised of the TRPP2 mutant and the TRPC3 or TRPC7 protein induce enhanced receptor-activated Ca(2+) influx that may lead to dysregulated cell growth in ADPKD. PMID- 19812036 TI - Double superhelix model of high density lipoprotein. AB - High density lipoprotein (HDL), the carrier of so-called "good" cholesterol, serves as the major athero-protective lipoprotein and has emerged as a key therapeutic target for cardiovascular disease. We applied small angle neutron scattering (SANS) with contrast variation and selective isotopic deuteration to the study of nascent HDL to obtain the low resolution structure in solution of the overall time-averaged conformation of apolipoprotein AI (apoA-I) versus the lipid (acyl chain) core of the particle. Remarkably, apoA-I is observed to possess an open helical shape that wraps around a central ellipsoidal lipid phase. Using the low resolution SANS shapes of the protein and lipid core as scaffolding, an all-atom computational model for the protein and lipid components of nascent HDL was developed by integrating complementary structural data from hydrogen/deuterium exchange mass spectrometry and previously published constraints from multiple biophysical techniques. Both SANS data and the new computational model, the double superhelix model, suggest an unexpected structural arrangement of protein and lipids of nascent HDL, an anti-parallel double superhelix wrapped around an ellipsoidal lipid phase. The protein and lipid organization in nascent HDL envisages a potential generalized mechanism for lipoprotein biogenesis and remodeling, biological processes critical to sterol and lipid transport, organismal energy metabolism, and innate immunity. PMID- 19812037 TI - The 26 S proteasome: from basic mechanisms to drug targeting. AB - The regulated degradation of proteins within eukaryotes and bacterial cells is catalyzed primarily by large multimeric proteases in ATP-dependent manner. In eukaryotes, the 26 S proteasome is essential for the rapid destruction of key regulatory proteins, such as cell cycle regulators and transcription factors, whose fast and tuned elimination is necessary for the proper control of the fundamental cell processes they regulate. In addition, the 26 S proteasome is responsible for cell quality control by eliminating defective proteins from the cytosol and endoplasmic reticulum. These defective proteins can be misfolded proteins, nascent prematurely terminated polypeptides, or proteins that fail to assemble into complexes. These diverse activities and its central role in apoptosis have made the proteasome an important target for drug development, in particular to combat malignancies. PMID- 19812038 TI - Phosphorylation of the par polarity complex protein Par3 at serine 962 is mediated by aurora a and regulates its function in neuronal polarity. AB - The Aurora kinases are a family of serine/threonine protein kinases that perform important functions during the cell cycle. Recently, it was shown that Drosophila Aurora A also regulates the asymmetric localization of Numb to the basal and the partitioning-defective (Par) complex to the apical cortex of neuroblasts by phosphorylating Par6. Here, we show that Aurora A is required for neuronal polarity. Suppression of Aurora A by RNA interference results in the loss of neuronal polarity. Aurora A interacts directly with the atypical protein kinase C binding domain of Par3 and phosphorylates it at serine 962. The phosphorylation of Par3 at serine 962 contributes to its function in the establishment of neuronal polarity. PMID- 19812039 TI - Role of the Rad52 amino-terminal DNA binding activity in DNA strand capture in homologous recombination. AB - Saccharomyces cerevisiae Rad52 protein promotes homologous recombination by nucleating the Rad51 recombinase onto replication protein A-coated single stranded DNA strands and also by directly annealing such strands. We show that the purified rad52-R70A mutant protein, with a compromised amino-terminal DNA binding domain, is capable of Rad51 delivery to DNA but is deficient in DNA annealing. Results from chromatin immunoprecipitation experiments find that rad52 R70A associates with DNA double-strand breaks and promotes recruitment of Rad51 as efficiently as wild-type Rad52. Analysis of gene conversion intermediates reveals that rad52-R70A cells can mediate DNA strand invasion but are unable to complete the recombination event. These results provide evidence that DNA binding by the evolutionarily conserved amino terminus of Rad52 is needed for the capture of the second DNA end during homologous recombination. PMID- 19812040 TI - Protein-tyrosine phosphatase alpha acts as an upstream regulator of Fyn signaling to promote oligodendrocyte differentiation and myelination. AB - The tyrosine kinase Fyn plays a key role in oligodendrocyte differentiation and myelination in the central nervous system, but the molecules responsible for regulating Fyn activation in these processes remain poorly defined. Here we show that receptor-like protein-tyrosine phosphatase alpha (PTPalpha) is an important positive regulator of Fyn activation and signaling that is required for the differentiation of oligodendrocyte progenitor cells (OPCs). PTPalpha is expressed in OPCs and is up-regulated during differentiation. We used two model systems to investigate the role of PTPalpha in OPC differentiation: the rat CG4 cell line where PTPalpha expression was silenced by small interfering RNA, and oligosphere derived primary OPCs isolated from wild-type and PTPalpha-null mouse embryos. In both cell systems, the ablation of PTPalpha inhibited differentiation and morphological changes that accompany this process. Although Fyn was activated upon induction of differentiation, the level of activation was severely reduced in cells lacking PTPalpha, as was the activation of Fyn effector molecules focal adhesion kinase, Rac1, and Cdc42, and inactivation of Rho. Interestingly, another downstream effector of Fyn, p190RhoGAP, which is responsible for Rho inactivation during differentiation, was not affected by PTPalpha ablation. In vivo studies revealed defective myelination in the PTPalpha(-/-) mouse brain. Together, our findings demonstrate that PTPalpha is a critical regulator of Fyn activation and of specific Fyn signaling events during differentiation, and is essential for promoting OPC differentiation and central nervous system myelination. PMID- 19812041 TI - Structural and biochemical evidence that a TEM-1 beta-lactamase N170G active site mutant acts via substrate-assisted catalysis. AB - TEM-1 beta-lactamase is the most common plasmid-encoded beta-lactamase in Gram negative bacteria and is a model class A enzyme. The active site of class A beta lactamases share several conserved residues including Ser(70), Glu(166), and Asn(170) that coordinate a hydrolytic water involved in deacylation. Unlike Ser(70) and Glu(166), the functional significance of residue Asn(170) is not well understood even though it forms hydrogen bonds with both Glu(166) and the hydrolytic water. The goal of this study was to examine the importance of Asn(170) for catalysis and substrate specificity of beta-lactam antibiotic hydrolysis. The codon for position 170 was randomized to create a library containing all 20 possible amino acids. The random library was introduced into Escherichia coli, and functional clones were selected on agar plates containing ampicillin. DNA sequencing of the functional clones revealed that only asparagine (wild type) and glycine at this position are consistent with wild-type function. The determination of kinetic parameters for several substrates revealed that the N170G mutant is very efficient at hydrolyzing substrates that contain a primary amine in the antibiotic R-group that would be close to the Asn(170) side chain in the acyl-intermediate. In addition, the x-ray structure of the N170G enzyme indicated that the position of an active site water important for deacylation is altered compared with the wild-type enzyme. Taken together, the results suggest the N170G TEM-1 enzyme hydrolyzes ampicillin efficiently because of substrate assisted catalysis where the primary amine of the ampicillin R-group positions the hydrolytic water and allows for efficient deacylation. PMID- 19812042 TI - Protein engineering of the quaternary sulfiredoxin.peroxiredoxin enzyme.substrate complex reveals the molecular basis for cysteine sulfinic acid phosphorylation. AB - Oxidative stress can damage the active site cysteine of the antioxidant enzyme peroxiredoxin (Prx) to the sulfinic acid form, Prx-SO(2)(-). This modification leads to inactivation. Sulfiredoxin (Srx) utilizes a unique ATP-Mg(2+)-dependent mechanism to repair the Prx molecule. Using selective protein engineering that involves disulfide bond formation and site-directed mutagenesis, a mimic of the enzyme.substrate complex has been trapped. Here, we present the 2.1 A crystal structure of human Srx in complex with PrxI, ATP, and Mg(2+). The Cys(52) sulfinic acid moiety was substituted by mutating this residue to Asp, leading to a replacement of the sulfur atom with a carbon atom. Because the Srx reaction cannot occur, the structural changes in the Prx active site that lead to the attack on ATP may be visualized. The local unfolding of the helix containing C52D resulted in the packing of Phe(50) in PrxI within a hydrophobic pocket of Srx. Importantly, this structural rearrangement positioned one of the oxygen atoms of Asp(52) within 4.3 A of the gamma-phosphate of ATP bound to Srx. These observations support a mechanism where phosphorylation of Prx-SO(2)(-) is the first chemical step. PMID- 19812043 TI - Targeting cancer vasculature via endoglin/CD105: a novel antibody-based diagnostic and therapeutic strategy in solid tumours. AB - Endoglin/CD105 is well acknowledged as being the most reliable marker of proliferation of endothelial cells, and it is overexpressed on tumour neovasculature. Our current knowledge of its structure, physiological role, and tissue distribution suggests that targeting of endoglin/CD105 is a novel and powerful diagnostic and therapeutic strategy in human malignancies, through the imaging of tumour-associated angiogenesis and the inhibition of endothelial cell functions related to tumour angiogenesis. Among biotherapeutic agents, monoclonal antibodies have shown a major impact on the clinical course of human malignancies of different histotypes. Along this line, the potential efficacy of anti endoglin/CD105 antibodies and their derivatives for clinical purposes in cancer is supported by a large body of available pre-clinical in vitro and in vivo data. In this review, the main findings supporting the translation of antibody-based endoglin/CD105 targeting from pre-clinical studies to clinical applications in human cancer are summarized and discussed. PMID- 19812044 TI - Vasoconstriction: tightening the noose through MMPs. PMID- 19812045 TI - The tail of Cx43: its crucial protective role in acute myocardial infarction. PMID- 19812046 TI - Is it time to start with device-based prognosticators? PMID- 19812047 TI - Feasibility and outcome of epicardial pulmonary vein isolation for lone atrial fibrillation using minimal invasive surgery and high intensity focused ultrasound. AB - AIMS: Transvenous pulmonary vein isolation (PVI) is the cornerstone of non pharmacological rhythm control therapy in symptomatic atrial fibrillation (AF). Success and complications rates are, however, still not optimal. New techniques and energy sources are therefore being developed. METHODS AND RESULTS: Fifteen patients with lone AF refractory for antiarrhythmic drugs (AADs) underwent PVI by minimal invasive epicardial off-pump monolateral right-sided video-assisted thoracic surgery (VATS) using the UltraCinch with high-intensity focused ultrasound (HIFU). Primary endpoint was successful ablation defined as absence of AF or atrial flutter/tachycardia after 6 months assessed by complaints, 12 lead electrocardiogram, and 96 h Holter monitoring. Secondary endpoints were ablation success at the end of follow-up irrespective of AADs use or re-ablation and complications related to the procedure. Mean age was 47 +/- 10 years and 14 (93%) were male. Eleven (73%) had paroxysmal, and 4 (27%) patients had persistent AF. Median AF history was 5 (1-12) years. At 6 months, six (40%) patients had sinus rhythm after one epicardial PVI (four on AADs). After 1.3 +/- 0.6 years, four (27%) patients had sinus rhythm after one epicardial PVI (two on AADs) and in six (40%) patients endocardial radiofrequency re-ablation was performed, which was successful in three patients (20%). Two patients (13%) were planned for re ablation. Three others (20%) refused re-ablation. Two major complications occurred (one late tamponade and one bleeding during surgery, necessitating sternotomy). CONCLUSION: Epicardial PVI using monolateral right-sided VATS with the UltraCinch delivering HIFU is feasible, but is associated with substantial complications. Furthermore, the success rate was low. More research is therefore warranted to assess optimal ablation techniques and energy sources to perform PVI. PMID- 19812048 TI - Right ventricular contractility as a measure of optimal interventricular pacing setting in cardiac resynchronization therapy. AB - AIMS: The aim of the present study was to assess whether right ventricular (RV) contractility can be used for optimization of the interventricular (VV) interval and to study the acute hemodynamic effect of different VV intervals on right and left ventricular (LV) contractility in patients referred for cardiac resynchronization therapy (CRT). METHODS AND RESULTS: Intracardiac LV and RV dP/dt were measured with a 0.014-in. sensor-tipped pressure guidewire during pacing at nine different VV intervals ranging from +80 ms (LV pre-excitation) to 80 ms (RV pre-excitation) in 26 patients who received a biventricular pacemaker. No correlation was found between the optimal VV intervals identified by maximum LV dP/dt and RV dP/dt, which were identical in only seven cases (27%). Only when testing slightly broader intervals (+/-20 ms) was there a statistically significant correlation (P= 0.037) between the optimized VV intervals. In the majority of patients (58%) either LV or RV pre-excitation was superior to simultaneous pacing according to LV dP/dt(max) measurements. CONCLUSION: RV dP/dt(max) failed to identify the optimal VV interval when compared with LV dP/dt(max) and can therefore not be recommended for VV optimization in CRT patients. PMID- 19812049 TI - Unsuccessful left ventricular lead implantation in two first-degree relatives. Is the coronary venous anatomy similar in both cases? AB - We reported on two unsuccessful implantations of the left ventricular lead in two first-degree relatives due to inability to cannulate the coronary sinus (CS). The anatomy of the coronary venous system investigated by means of dual source computed tomography showed several similarities in both patients: narrowing of the proximal part of CS and a small number of CS tributaries. PMID- 19812050 TI - Implantable cardioverter defibrillator therapy in chronic kidney disease: a meta analysis. AB - AIMS: Recent observational studies have shown that implantable cardioverter defibrillator (ICD) patients with chronic kidney disease (CKD) have increased mortality and therefore the value of device therapy in this group has been questioned. The purpose of this meta-analysis was to systematically analyse the effect of renal dysfunction on mortality of ICD patients. METHODS AND RESULTS: Pubmed, Cochrane clinical trials database, and EMBASE were searched until December 2008. In addition, a manual search was performed using review articles, reference lists of papers, and abstracts from conference reports. Of the 90 initially identified studies, 11 observational studies with 3010 patients were analysed. The meta-analysis of these studies showed that CKD was associated with higher mortality risk (HR = 3.44, 95% CI 2.82-4.21, Z = 12.09, P < 0.001) while there were no significant differences between individual trials (P = 0.09, I(2) = 37.8%). A subgroup analysis which included the four studies that used estimated glomerular filtration rate (GFR) <60 mL/min/1.73 m(2) to define CKD showed a higher mortality in the CKD group as well (HR = 3.06, 95% CI 2.31-4.04, Z = 7.84, P < 0.001) without significant heterogeneity (P = 0.38, I(2) = 5.2%). CONCLUSION: Our meta-analysis suggests that CKD is associated with increased mortality in patients who receive ICD therapy. Undoubtedly, prospective studies are needed in order to elucidate the impact of different stages of CKD in this setting. Given that the CKD prevalence is rapidly increasing, there is an imperative need for better risk stratification of ICD therapy candidates. PMID- 19812051 TI - Thromboaspiration of left atrial clot during ablation of atrial fibrillation. AB - We describe a case during which a left atrial thrombus was visualized within the left atrium attached to a circular catheter during an atrial fibrillation ablation procedure. This was managed by successful thromboaspiration using a steerable sheath, preventing a potential serious complication. PMID- 19812052 TI - Marked accumulation of 27-hydroxycholesterol in SPG5 patients with hereditary spastic paresis. AB - Patients with a recessively inherited "pure" hereditary spastic paresis (SPG5) have mutations in the gene coding for the oxysterol 7 alpha hydroxylase (CYP7B1). One of the expected metabolic consequences of such mutations is accumulation of oxysterol substrates due to decreased enzyme activity. In accordance with this, we demonstrate here that four patients with the SPG5 disease have 6- to 9-fold increased plasma levels of 27-hydroxycholesterol. A much higher increase, 30- to 50-fold, was found in cerebrospinal fluid. The plasma levels of 25 hydroxycholesterol were increased about 100-fold. There were no measurable levels of this oxysterol in cerebrospinal fluid. The pattern of bile acids in serum was normal, suggesting a normal bile acid synthesis. The findings are discussed in relation to two transgenic mouse models with increased levels of 27-hydroxy cholesterol in the circulation but without neurological symptoms: the cyp27a1 transgenic mouse and the cyp7b1 knockout mouse. The absolute plasma levels of 27 hydroxycholesterol in the latter models are, however, only about 20% of those in the SPG5 patients. If the accumulation of 27-hydroxycholesterol is an important pathogenetic factor, a reduction of its levels may reduce or prevent the neurological symptoms. A possible strategy to achieve this is discussed. PMID- 19812053 TI - APOC1 T45S polymorphism is associated with reduced obesity indices and lower plasma concentrations of leptin and apolipoprotein C-I in aboriginal Canadians. AB - Apolipoprotein (apo) C-I is a constituent of chylomicrons, very low density lipoprotein, and high density lipoprotein. The role of apo C-I in human metabolism is incompletely defined. We took advantage of a naturally occurring amino acid polymorphism that is present in aboriginal North Americans, namely apo C-I T45S. We assessed the hypothesis that metabolic traits, including obesity related and lipoprotein-related traits, would differ between carriers and noncarriers of apo C-I T45S. A genotyping assay was developed for APOC1 T45S and genotypes were determined in a sample of 410 Canadian Oji-Cree subjects. The allele frequency of the apo C-I S45 allele was approximately 8% in this sample. We observed the apo C-I S45 allele was significantly associated with 1) lower percent body fat (P < 0.05), 2) lower waist circumference (P = 0.058), 3) lower serum leptin levels (P < 0.05), and 4) lower plasma apo C-I levels (P < 0.0001), using a newly developed ELISA-based method. Taken together, these results suggest that at the whole human phenotype level, apo C-I is associated with the complex metabolic trait of obesity as well as with serum leptin levels. PMID- 19812054 TI - Adjusting for clinical covariates improves the ability of B-type natriuretic peptide to distinguish cardiac from non-cardiac dyspnoea: a sub-study of HEARD IT. AB - AIMS: We sought to create a model that adjusts B-type natriuretic peptide (BNP) for specific covariates to better distinguish cardiac from non-cardiac dyspnoea. METHODS AND RESULTS: HEARD-IT was a multicentre, prospective study of the diagnostic utility of acoustic cardiography in the emergency department. Dyspnoeic patients more than 40 years were eligible. Two cardiologists independently adjudicated the HF outcome. Using logistic regression, a model adjusting BNP for pertinent covariates was developed (n = 740). The mean age was 66 +/- 13 years. Age, gender, ethnicity, body mass index, blood urea nitrogen, and creatinine affected BNP levels independently of HF. The model adjusting BNP for these covariates improved the area under receiver operator characteristic curve for HF compared with BNP alone (0.948, 95% CI 0.934-0.963 vs. 0.937, 95% CI 0.920-0.954; P = 0.004). Net reclassification improvement, a novel metric of model performance, was 3.5% for those without HF (P = 0.05) compared with conventional, unadjusted BNP cut-offs. Thirteen of 116 (11%) patients without HF, but with unadjusted BNP values > or =100 pg/mL, were correctly reclassified as not having HF with the adjusted BNP model. CONCLUSION: Adjusting BNP for important covariates may improve its ability to distinguish cardiac from non cardiac dyspnoea. PMID- 19812059 TI - Early but not late stent thrombosis is influenced by residual platelet aggregation in patients undergoing coronary interventions. AB - AIMS: Recent studies suggest a relevant association of post-interventional residual platelet aggregation (RPA) under therapy with oral platelet inhibitors and the occurrence of atherothrombotic events. The influence of post interventional RPA on the incidence of stent thrombosis (ST) has not been sufficiently evaluated in consecutive unselected cohorts of percutaneous coronary intervention (PCI) patients. The aim of this observational study was to investigate the impact of RPA on the incidence of ST within 3 months in patients treated with dual antiplatelet therapy. METHODS AND RESULTS: The study population included a consecutive cohort of 1019 patients treated with PCI [n = 741 bare metal stent (BMS) and n = 278 drug-eluting stent (DES)] due to symptomatic coronary artery disease. Residual platelet activity was assessed by adenosine disphosphate (20 micromol/L)-induced PA after 600 mg clopidogrel loading dose. Maximum RPA was measured as peak of aggregation, final RPA was measured 5 min after addition of agonist. The primary endpoint was the occurrence of ST within 3 months defined according to academic research consortium (ARC) criteria. Final and maximum RPA were independent predictors of ST after 3 months. In secondary analysis, the observed effects were independently associated with early ST (HR 1.05, 95% CI 1.01-1.08 and HR 1.05, 95% CI 1.01-1.09, P < 0.01, respectively). However, incidence of 3-month late stent thrombosis (LAT) was not influenced by post-interventional RPA in multivariable analysis. CONCLUSION: Post interventional RPA is associated with the occurrence of early ST in patients treated with either BMS or DES; however, there is no predictive value of RPA for the incidence of 3-month LAT, suggesting the involvement of other possible mechanisms like discontinuation of antiplatelet therapy. PMID- 19812060 TI - Impacts of acute severe pulmonary regurgitation on right ventricular geometry and contractility assessed by tissue-Doppler echocardiography. AB - AIMS: Little is known of the impact of acute right ventricular (RV) volume overload on RV function. We assessed the impact of acute severe pulmonary regurgitation (PR) on global and regional RV function by applying novel quantitative echocardiographic markers of myocardial performance in an animal model. METHODS AND RESULTS: Transthoracic echocardiography, including tissue Doppler echocardiography for the evaluation of regional longitudinal function, was performed immediately before and after induction of severe PR by deployment of a stent in the pulmonary valve annulus of 32 farm pigs. Acute PR was associated with significant changes in RV geometry illustrated by an increase in RV diameter and area by 22 and 32%, respectively, P < 0.001 for both, and the eccentricity index increased by 21% in end-diastole, P < 0.0001. RV radial function as assessed by RV short-axis fractional shortening increased by 18%, P = 0.03, whereas other measures of RV ejection fraction by longitudinal function remained unchanged. There were no changes in the longitudinal basal myocardial isovolumic acceleration, peak systolic velocity, strain rate, or strain. CONCLUSION: The RV seems to accommodate well to acute severe PR. No changes in global or regional longitudinal contractility or deformation were observed despite significant changes in the cardiac chamber geometry. An increase in radial shortening may imply that the RV compensates by increasing radial contraction as an adjunct to dilatation. PMID- 19812061 TI - Second-line chemotherapy with biweekly paclitaxel after failure of fluoropyrimidine-based treatment in patients with advanced or recurrent gastric cancer: a report from the gastrointestinal oncology group of the Tokyo cooperative oncology group, TCOG GC-0501 trial. AB - OBJECTIVE: A multicenter trial was conducted to evaluate the efficacy and safety of paclitaxel every 2 weeks in patients with advanced or recurrent gastric cancer who had previously received fluoropyrimidine-based chemotherapy. METHODS: The subjects were patients with gastric cancer who had disease progression or recurrence while receiving fluoropyrimidine-based chemotherapy. All patients had adequate major organ functions with an Eastern Cooperative Oncology Group performance status (PS) of 0-2. Paclitaxel 140 mg/m(2) was administered intravenously on days 1 and 15 of a 4-week cycle. The primary endpoint was the response rate. Secondary endpoints were progression-free survival (PFS), overall survival and safety. RESULTS: Response was assessable in 40 of 41 enrolled patients. Their median age was 63 (range: 48-77) years, and PS was 0 in 22 patients, 1 in 13 and 2 in 5. Previous treatment included S-1 (1 M tegafur-0.4 M gimestat-1 M otastat potassium) monotherapy in 32 patients and S-1-based combination therapy in 5. The median number of administered courses of paclitaxel was 3.5 (1-14). The response rate was 17.5% (95% confidence interval: 7.3-32.8%, partial response: 7, stable disease: 21, progressive disease: 10 and not evaluable: 2). The disease control rate was 70.0%, the median PFS was 111 days and the median overall survival was 254 days. Major adverse events of Grade 3 or 4 were neutropenia (27.5%), anemia (12.5%), diarrhea (2.5%) and sensory neuropathy (2.5%). CONCLUSIONS: Biweekly paclitaxel seemed to be one of the useful chemotherapies after failure of fluoropyrimidine-based treatment in patients with advanced or recurrent gastric cancer. PMID- 19812062 TI - Radiation-induced parotid gland changes in oral cancer patients: correlation between parotid volume and saliva production. AB - OBJECTIVE: To evaluate whether saliva production reflects the parotid volume during the course of radiation therapy (RT) in patients with head-and-neck cancer. METHODS: Twenty patients with advanced oral squamous cell carcinomas, who were treated with preoperative chemo-RT, underwent morphological assessment with CT or MRI and functional assessment with the Saxon test. For the Saxon test, saliva production was measured by weighing a gauze pad before and 2 min after chewing without swallowing; the low-normal value is 2 g. Saliva production and parotid volumes before and 2 weeks after RT were compared with the paired t-test, the Spearman rank correlation test and the Fisher exact test. RESULTS: After 30 Gy irradiation, mean saliva production was decreased from 4.2 to 1.0 g (P < 0.01); the reduction in saliva production ranged from 1.7 to 5.4 g (mean 3.2 g). The mean parotid volume was decreased from 68.2 to 47.9 cm(3) (P < 0.01); the post-RT:pre-RT parotid volume ratio ranged from 54% to 85% (mean 71%). Although the initial parotid ;volume was correlated with initial saliva production (r = 0.47, P = 0.04), no significant correlation was noted after RT (r = 0.08, P = 0.71), and there were considerable individual variations. The parotid volume ratio was inversely correlated with the saliva-reduction amount (r = - 0.79, P < 0.01). CONCLUSIONS: There was a correlation between decreased parotid gland volume and decreased saliva production in patients with head-and-neck cancer undergoing RT. Parotid volume reduction may predict parotid gland function. PMID- 19812063 TI - Arabidopsis replication protein A 70a is required for DNA damage response and telomere length homeostasis. AB - Replication protein A1 (RPA1/RPA70) forms a heterotrimeric complex together with RPA2/RPA32 and RPA3/RPA14 subunits which plays essential roles in various aspects of DNA metabolism including replication, repair, recombination and telomere maintenance. Compared with RPA70 in yeast and mammals, limited information is available about the factor in plants. In this study, we analyzed the functions of AtRPA70a, which is most similar to human RPA70 among four paralogs in Arabidopsis thaliana. RNA blot analysis showed that AtRPA70a is expressed ubiquitously in plant organs containing differentiated and meristematic tissues, while its expression was up-regulated in response to DNA damage stress. Yeast two-hybrid and co-immunoprecipitation analyses showed that AtRPA70a interacted preferentially with Arabidopsis RPA32a, one of two paralogs. Inactivation of AtRPA70a by T-DNA insertion did not affect growth under normal conditions, but resulted in increased sensitivity to genotoxic agents such as methylmethane sulfonate, bleomycin and hydroxyurea. Terminal restriction fragment analysis revealed that telomere lengths in an AtRPA70a-deficient line were significantly larger than in the wild type, whereas those in the mutant expressing antisense AtTERT (telomerase catalytic subunit gene) were shortened during successive generations. These results demonstrate that AtRPA70a is involved in repair of double-strand DNA breaks and possibly contributes to telomerase-dependent telomere length regulation. PMID- 19812064 TI - Rice BRITTLE CULM 5 (BRITTLE NODE) is involved in secondary cell wall formation in the sclerenchyma tissue of nodes. AB - Several brittle culm (bc) mutants known in grasses are considered excellent materials to study the process of secondary cell wall formation. The brittle phenotype of the rice bc5 (brittle node) mutant appears exclusively in the developed nodes, which is distinct from other bc mutants (bc1, 2, 3, 4, 6 and 7) that show the brittle phenotype in culms and leaves. To address the defects of the rice bc5 mutant in node-specific cell wall formation, we analyzed tissue morphology and cell wall composition. The bc5 mutation was found to affect the cell wall deposition of node sclerenchyma tissues at 1 week after heading, the stage at which the cell wall sugar content is reduced, in the bc5 nodes, compared with wild-type nodes. Moreover, decreased accumulation of lignin and thickness of cell walls in the sclerenchyma tissues were also observed in the bc5 nodes. The amounts of cellulose and hemicellulose were reduced to 53 and 65% of those in the wild-type plants, respectively. Sugar composition and glycosidic linkage analyses of the hemicellulose showed that the accumulation of glucuronosyl arabinoxylan in bc5 nodes was perturbed by the mutation. The bc5 locus was narrowed to an approximately 3.1 Mb region of chromosome 2, where none of the other bc genes is located. The bc5 mutation appeared to reduce the expression levels of the OsCesA genes in the nodes after heading. The results indicate that the BC5 gene regulates the development of secondary cell walls of node sclerenchyma tissues. PMID- 19812065 TI - Pharmacokinetics of nevirapine in HIV-infected children with and without malnutrition receiving divided adult fixed-dose combination tablets. AB - OBJECTIVES: To determine the relationship between nutritional status and nevirapine exposure by comparing the pharmacokinetics of nevirapine in HIV infected children of different ages with and without malnutrition receiving divided tablets of Triomune 30 (stavudine + lamivudine + nevirapine) in accordance with Malawi National Guidelines. METHODS: Children were recruited in weight-based dosage bands and nutritional status classified according to weight for height. Total and unbound plasma nevirapine concentrations were measured over a full dosing interval. Multivariate linear and logistic regression analyses were performed to determine the effects of malnutrition, age, dose and other factors on nevirapine exposure and likelihood of achieving therapeutic nevirapine trough concentrations. RESULTS: Forty-three children were recruited (37 included for analysis). Mild to moderate malnutrition was present in 12 (32%) children; 25 (68%) were of normal nutritional status. There was no effect of malnutrition on any measure of total drug exposure or on the unbound fraction of nevirapine. Nevirapine exposure was strongly related to dose administered (P = 0.039) and to age (for every yearly increase in age there was an approximately 88% increase in the odds of achieving a therapeutic nevirapine concentration; P = 0.056, 95% confidence interval 0.983-3.585). CONCLUSIONS: Use of divided adult Triomune 30 tablets in treating young children results in significant underdosing. No independent effect of malnutrition on total and unbound nevirapine exposures was observed. These data support the use of bespoke paediatric antiretroviral formulations. PMID- 19812067 TI - Developmental morphology of strap-shaped gametophytes of Colysis decurrens: a new look at meristem development and function in fern gametophytes. AB - BACKGROUND AND AIMS: The gametophytes of most homosporous ferns are cordate thalloid in shape. Some are strap- or ribbon-shaped and have been assumed to have evolved from terrestrial cordate shapes as an adaptation to epiphytic habitats. The aim of the present study was to clarify the morphological evolution of the strap-shaped gametophyte of microsoroids (Polypodiaceae) by precise analysis of their development. METHODS: Spores of Colysis decurrens collected in Kagoshima, Japan, were cultured and observed microscopically. Epi-illuminated micrographs of growing gametophytes were captured every 24 h, allowing analysis of the cell lineage of meristems. Light microscopy of resin-sections and scanning electron microscopy were also used. KEY RESULTS: Contrary to previous assumptions that strap-shaped Colysis gametophytes have no organized meristem, three different types of meristems are formed during development: (1) apical-cell based - responsible for early growth; (2) marginal - further growth, including gametophyte branching; and (3) multicellular - formation of cushions with archegonia. The cushion is two or three layers thick and intermittent. The apical cell and multicellular meristems are similar to those of cordate gametophytes of other ferns, but the marginal meristem is unique to the strap-shaped gametophyte of this fern. CONCLUSIONS: The strap-shaped gametophytes of C. decurrens may have evolved from ancestors with a cordate shape by insertion of the marginal meristem phase between the first apical-cell-based meristem and subsequent multicellular meristem phases. Repeated retrieval of the marginal meristem at the multicellular meristem phase would result in indefinite prolongation of gametophyte growth, an ecological adaptation to epiphytic habitats. PMID- 19812066 TI - Influence of CYP2B6 polymorphisms on the persistence of plasma nevirapine concentrations following a single intra-partum dose for the prevention of mother to child transmission in HIV-infected Thai women. AB - OBJECTIVES: To investigate the association of single nucleotide polymorphisms (SNPs) with nevirapine concentrations following intra-partum single-dose nevirapine. METHODS: Plasma and DNA samples were obtained from 330 HIV-infected Thai women who received intra-partum single-dose nevirapine in the PHPT-2 clinical trial to prevent perinatal HIV transmission. Nine SNPs within CYP2B6, CYP3A4 and ABCB1 were genotyped by real-time PCR. Nevirapine plasma concentrations were determined by HPLC and used in a population pharmacokinetic analysis. RESULTS: Higher nevirapine exposure was observed in women carrying the CYP2B6 516G>T polymorphism, but this did not reach statistical significance (P = 0.054). The TGATC CYP2B6 haplotype (g.3003T, 516G, 785A, g.18492T and g.21563C) was associated with increased nevirapine clearance and lower exposure (P = 0.0029). The median time for nevirapine concentrations to reach 10 ng/mL post partum (nevirapine IC(50) for HIV-1) was 14 days [interquartile range (IQR, 14 18)] for TGATC homozygotes, 16 days (14-20) for TGATC heterozygotes and 18 days (14-20) for non-TGATC homozygotes (P = 0.020). CONCLUSIONS: The CYP2B6 516G>T impact on nevirapine concentrations was less pronounced after intra-partum single dose nevirapine than reported under steady-state conditions, perhaps due to lack of enzyme auto-induction at the time of dosing. Although the TGATC CYP2B6 haplotype may shorten the persistence of nevirapine post-partum, its practical implications for the prevention of HIV transmission or selection of resistance mutations are likely limited. PMID- 19812068 TI - Live birth among Iguanian lizards predates Pliocene--Pleistocene glaciations. AB - Among tetrapods, viviparity is estimated to have evolved independently within Squamata (lizards and snakes) more than 100 times, most frequently in species occupying cold climate environments. Because of this relationship with cold climates, it is sometimes assumed that many origins of squamate viviparity occurred over the past 2.5-4 Myr during the Pliocene-Pleistocene glaciations; however, this hypothesis is untested. Divergence-dating analysis on a 733-species tree of Iguanian lizards recovers 20 independent lineages that have evolved viviparity, of which 13 multispecies groups derived live birth prior to glacial advances (8-66 Myr ago). These results place the transitions from egg-laying to live birth among squamates in a well-supported historical context to facilitate examination of the underlying phenotypic and genetic changes associated with this complex shift in reproduction. PMID- 19812069 TI - Complex vocal imitation during ontogeny in a bat. AB - Vocal imitation--the ability to learn a previously unknown acoustic signal from a tutor--is considered to be a key innovation in the evolution of speech. This faculty is very rare and patchily distributed within the animal kingdom, suggesting multiple instances of convergent evolution. It has long been predicted that bats should be capable of vocal imitation and our results provide evidence for this phenomenon. We report that pups of the bat Saccopteryx bilineata learn a complex vocalization through vocal imitation. During ontogeny, pups of both sexes imitate territorial song from adult males, starting with simple precursor songs that develop into genuine renditions. The resemblance of pup renditions to their acoustic model is not caused by physical maturation effects, is independent of pups' gender and relatedness towards adult males and becomes more pronounced during ontogeny, showing that auditory experience is essential for vocal development. Our findings indicate that the faculty of vocal imitation is more widespread than previously thought and emphasize the importance of research on audiovocal communication in bats for a better understanding of the evolutionary origin of vocal imitation. PMID- 19812070 TI - Ovulation mode modifies paternity monopolization in mammals. AB - There are two forms of ovulation: spontaneous and induced. As copulation triggers ovulation for induced ovulators, males can predict the timing of ovulation and may have greater paternity monopolization than spontaneous ovulators. However, this prediction has never, to my knowledge, been tested. Using a cross-species comparison I examined the percentage of offspring sired within a litter (single paternity) and in social species the percentage of offspring sired by the dominant male (alpha paternity). My results indicate that ovulation mode alters the ability of males to monopolize paternity, with males of induced ovulators having higher single paternity and greater alpha paternity where male-female association is intermittent. PMID- 19812071 TI - Relationships between specific surface area and pore size in electrospun polymer fibre networks. AB - From consideration of the extent of contact between fibres in electrospun polymer networks, we provide theory relating the specific surface area of the network to the characteristic dimensions of interfibre voids. We show that these properties are strongly influenced by the cross-sectional morphologies of fibres. Whereas porosity has a strong influence on pore dimensions, in the range of porosities typically obtained in real networks, its influence on specific surface area is weak. By considering reference geometries of collapsed ribbons and fibres with circular cross sections, we demonstrate that at a given network porosity, fibre parameters that increase the specific surface area reduce the characteristic dimensions of voids. The implications of the theory, mainly in the context of cell proliferation on electrospun polymer scaffolds, are discussed; the theory has relevance also to future applications of these materials in composites. PMID- 19812072 TI - Mathematical models for assessing the role of airflow on the risk of airborne infection in hospital wards. AB - Understanding the risk of airborne transmission can provide important information for designing safe healthcare environments with an appropriate level of environmental control for mitigating risks. The most common approach for assessing risk is to use the Wells-Riley equation to relate infectious cases to human and environmental parameters. While it is a simple model that can yield valuable information, the model used as in its original presentation has a number of limitations. This paper reviews recent developments addressing some of the limitations including coupling with epidemic models to evaluate the wider impact of control measures on disease progression, linking with zonal ventilation or computational fluid dynamics simulations to deal with imperfect mixing in real environments and recent work on dose-response modelling to simulate the interaction between pathogens and the host. A stochastic version of the Wells Riley model is presented that allows consideration of the effects of small populations relevant in healthcare settings and it is demonstrated how this can be linked to a simple zonal ventilation model to simulate the influence of proximity to an infector. The results show how neglecting the stochastic effects present in a real situation could underestimate the risk by 15 per cent or more and that the number and rate of new infections between connected spaces is strongly dependent on the airflow. Results also indicate the potential danger of using fully mixed models for future risk assessments, with quanta values derived from such cases less than half the actual source value. PMID- 19812073 TI - Exhaled droplets due to talking and coughing. AB - Respiratory infections can be spread via 'contact' with droplets from expiratory activities such as talking, coughing and sneezing, and also from aerosol generating clinical procedures. Droplet sizes predominately determine the times they can remain airborne, the possibility of spread of infectious diseases and thus the strategies for controlling the infections. While significant inconsistencies exist between the existing measured data on respiratory droplets generated during expiratory activities, a food dye was used in the mouth during measurements of large droplets, which made the expiratory activities 'unnatural'. We carried out a series of experiments using glass slides and a microscope as well as an aerosol spectrometer to measure the number and size of respiratory droplets produced from the mouth of healthy individuals during talking and coughing with and without a food dye. The total mass of respiratory droplets was measured using a mask, plastic bag with tissue and an electronic balance with a high precision. Considerable subject variability was observed and the average size of droplets captured using glass slides and microscope was about 50-100 microm. Smaller droplets were also detected by the aerosol spectrometer. More droplets seemed to be generated when a food dye was used. PMID- 19812074 TI - Personalized ventilation as a control measure for airborne transmissible disease spread. AB - The protective role of personalized ventilation (PV) against plausible airborne transmissible disease was investigated using cough droplets released from a 'coughing machine' simulating the human cough at different distances (1, 1.75 and 3 m) from the PV user. Particle image velocimetry was used to characterize and visualize the interaction between the cough-generated multiphase flow and PV induced flow in the inhalation zone of the thermal breathing manikin. A dose response model for unsteady imperfectly mixed environment was used to estimate the reduction in infection risk of two common diseases that can be transmitted by airborne mode. PV was able to both reduce the peak aerosol concentration levels and shorten the exposure time at all the examined injection distances. PV could reduce the infection risks of two diseases, influenza A and tuberculosis, by between 27 and 65 per cent. The protection offered by PV is less effective at a distance of 1.75 m than the other distances, as shown in the risk assessment results, as the PV-generated flow was blown off by the cough-generated flow for the longest time. Results of this study demonstrate the ability of desktop PV to mitigate the infection risk of airborne transmissible disease. PMID- 19812075 TI - Conservative ecological and evolutionary patterns in liverwort-fungal symbioses. AB - Liverworts, the most ancient group of land plants, form a range of intimate associations with fungi that may be analogous to the mycorrhizas of vascular plants. Most thalloid liverworts contain arbuscular mycorrhizal glomeromycete fungi similar to most vascular plants. In contrast, a range of leafy liverwort genera and one simple thalloid liverwort family (the Aneuraceae) have switched to basidiomycete fungi. These liverwort switches away from glomeromycete fungi may be expected to parallel switches undergone by vascular plants that target diverse lineages of basidiomycete fungi to form ectomycorrhizas. To test this hypothesis, we used a cultivation-independent approach to examine the basidiomycete fungi associated with liverworts in varied worldwide locations by generating fungal DNA sequence data from over 200 field collections of over 30 species. Here we show that eight leafy liverwort genera predominantly and consistently associate with members of the Sebacina vermifera species complex and that Aneuraceae thalloid liverworts associate nearly exclusively with Tulasnella species. Furthermore, within sites where multiple liverwort species co-occur, they almost never share the same fungi. Our analyses reveal a strikingly conservative ecological and evolutionary pattern of liverwort symbioses with basidiomycete fungi that is unlike that of vascular plant mycorrhizas. PMID- 19812076 TI - Sigma viruses from three species of Drosophila form a major new clade in the rhabdovirus phylogeny. AB - The sigma virus (DMelSV), which is a natural pathogen of Drosophila melanogaster, is the only Drosophila-specific rhabdovirus that has been described. We have discovered two new rhabdoviruses, D. obscura and D. affinis, which we have named DObsSV and DAffSV, respectively. We sequenced the complete genomes of DObsSV and DMelSV, and the L gene from DAffSV. Combining these data with sequences from a wide range of other rhabdoviruses, we found that the three sigma viruses form a distinct clade which is a sister group to the Dimarhabdovirus supergroup, and the high levels of divergence between these viruses suggest that they deserve to be recognized as a new genus. Furthermore, our analysis produced the most robustly supported phylogeny of the Rhabdoviridae to date, allowing us to reconstruct the major transitions that have occurred during the evolution of the family. Our data suggest that the bias towards research into plants and vertebrates means that much of the diversity of rhabdoviruses has been missed, and rhabdoviruses may be common pathogens of insects. PMID- 19812077 TI - Words as alleles: connecting language evolution with Bayesian learners to models of genetic drift. AB - Scientists studying how languages change over time often make an analogy between biological and cultural evolution, with words or grammars behaving like traits subject to natural selection. Recent work has exploited this analogy by using models of biological evolution to explain the properties of languages and other cultural artefacts. However, the mechanisms of biological and cultural evolution are very different: biological traits are passed between generations by genes, while languages and concepts are transmitted through learning. Here we show that these different mechanisms can have the same results, demonstrating that the transmission of frequency distributions over variants of linguistic forms by Bayesian learners is equivalent to the Wright-Fisher model of genetic drift. This simple learning mechanism thus provides a justification for the use of models of genetic drift in studying language evolution. In addition to providing an explicit connection between biological and cultural evolution, this allows us to define a 'neutral' model that indicates how languages can change in the absence of selection at the level of linguistic variants. We demonstrate that this neutral model can account for three phenomena: the s-shaped curve of language change, the distribution of word frequencies, and the relationship between word frequencies and extinction rates. PMID- 19812078 TI - Long-term changes in metapopulation genetic structure: a quarter-century retrospective study on low-Arctic rock pool Daphnia. AB - Population genetic surveys approximately 25 years apart examined the distribution and abundance of asexual clones of the freshwater zooplankter Daphnia pulex complex in rock pools near Churchill, Manitoba, Canada. In 1984-1985, melanic members of this species complex were present in 131 rock pools at this site, but were only detected in 90 of these pools in 2007-2008. Allozymic surveys conducted during these two time periods revealed that 59 per cent of these populations showed unchanged clonal composition. Total clonal replacement occurred in 8 per cent of the populations, while the others (33%) included a mixture of 'resident' clones and new 'colonists'. We discuss these changes in light of shifts in biotic and abiotic factors. We also discuss the use of rock pool habitats as 'sentinel' systems for examining long-term environmental changes in the ecological genetics of aquatic organisms in the Arctic. PMID- 19812079 TI - Female reproductive strategies in orangutans, evidence for female choice and counterstrategies to infanticide in a species with frequent sexual coercion. AB - Intersexual conflicts over mating can engender antagonistic coevolution of strategies, such as coercion by males and selective resistance by females. Orangutans are exceptional among mammals for their high levels of forced copulation. This has typically been viewed as an alternative mating tactic used by the competitively disadvantaged unflanged male morph, with little understanding of how female strategies may have shaped and responded to this behaviour. Here, we show that male morph is not by itself a good predictor of mating dynamics in wild Bornean orangutans but that female conception risk mediated the occurrence and quality of male-female interactions. Near ovulation, females mated cooperatively only with prime flanged males who they encountered at higher rates. When conception risk was low, willingness to associate and mate with non-prime males increased. Our results support the hypothesis that, together with concealed ovulation, facultative association is a mechanism of female choice in a species in which females can rarely avoid coercive mating attempts. Female resistance, which reduced copulation time, may provide an additional mechanism for mate selection. However, coercive factors were also important as prime males were frequently aggressive to females and females used mating strategies consistent with infanticide avoidance. PMID- 19812080 TI - Recent advances in Chinese palaeontology. AB - Discoveries are a driving force for progress in palaeontology. Palaeontology as a discipline of scientific inquiry has gained many fresh insights into the history of life, from the discoveries of many new fossils in China in the last 20 years, and from the new ideas derived from these fossils. This special issue of Proceedings of Royal Society B entitled Recent Advances in Chinese Palaeontology selects some of the very latest studies aimed at resolving the current problems of palaeontology and evolutionary biology based on new fossils from China. These fossils and their studies help to clarify some historical debates about a particular fossil group, or to raise new questions about history of life, or to pose a new challenge in our pursuit of science. These works on new Chinese fossils have covered the whole range of the diversity through the entire Phanerozoic fossil record. PMID- 19812081 TI - Attention focusing in a sit-and-wait forager: a spider controls its prey detection ability in different web sectors by adjusting thread tension. AB - Focusing attention is a way for animals to search for and to obtain food efficiently. This study examines whether a sit-and-wait forager, the orb-web spider Cyclosa octotuberculata, focuses its attention on limited foraging areas. Video records of foraging activity revealed that the spiders detected prey trapped in the west and east sectors of their web less frequently than prey trapped in the north and south sectors. Comparison of photos of the web hub area with and without spiders present revealed that the spiders pulled radii towards the centre when waiting for prey. Radius pulling is stronger in the north and south web sectors than in the west and east sectors, possibly causing more tension in radii running vertically. Experimental manipulation indicated that the spiders responded to prey quicker when thread tension was increased. The results suggest that C. octotuberculata focus their attention on the web areas above and below the spider by adjusting the tension in web threads; and this causes higher prey detection rates in these areas. PMID- 19812082 TI - Activational effects of odours on avian navigation. AB - The sensory basis of the navigational map remains one of the most important and intriguing questions in animal behaviour. In birds, odours have been hypothesized to provide the primary source of map information. Convincing tests have shown that experienced homing pigeons rely on map information obtained at sites where they are exposed to natural odours, even if subsequently released (without additional olfactory information) at a different site. These findings have been interpreted as support for the olfactory map hypothesis. Using this 'false release-site' (FRS) approach, we compared the effects of exposure to natural odours with that of exposure to a series of artificial odours lacking spatial information. Our findings show that olfactory exposure to either natural or artificial odours at an FRS caused pigeons to rely on map information obtained at the FRS, even if subsequently released at the true-release site in the opposite direction from the home loft. Because artificial odours did not provide map information, however, the findings clearly demonstrate that olfactory exposure provides no navigational information to pigeons whatsoever; instead it activates an independent non-olfactory map system. This test decisively contradicts the olfactory map hypothesis, which predicts that olfactory cues are the primary source of navigational information used by birds. PMID- 19812083 TI - Rooks perceive support relations similar to six-month-old babies. AB - Some corvids have demonstrated cognitive abilities that rival or exceed those of the great apes; for example, tool use in New Caledonian crows, and social cognition, episodic-like memory and future planning in Western scrub-jays. Rooks appear to be able to solve novel tasks through causal reasoning rather than simple trial-and-error learning. Animals with certain expectations about how objects interact would be able to narrow the field of candidate causes substantially, because some causes are simply 'impossible'. Here we present evidence that rooks hold such expectations and appear to possess perceptual understanding of support relations similar to that demonstrated by human babies, which is more comprehensive than that of chimpanzees. PMID- 19812084 TI - Low-quality females prefer low-quality males when choosing a mate. AB - Mate choice studies routinely assume female preferences for indicators of high quality in males but rarely consider developmental causes of within-population variation in mating preferences. By contrast, recent mate choice models assume that costs and benefits of searching or competing for high-quality males depend on females' phenotypic quality. A prediction following from these models is that manipulation of female quality should alter her choosiness or even the direction of her mating preferences. We here provide (to our knowledge) the first example where an experimental manipulation of female quality induced a mating preference for low-quality males. Zebra finches (Taeniopygia guttata) reared in small or large experimental broods became high- or low-quality adults, respectively. Only high-quality females preferred high-quality males' mate-advertising songs, while all low-quality females preferred low-quality males' song. Subsequent breeding trials confirmed this pattern: latency until egg laying was shortest in quality matched pairs, indicating that quality-matched birds were accepted faster as partners. Females produced larger eggs when mated with high-quality males, regardless of their own quality, indicating consensus regarding male quality despite the expression of different choices. Our results demonstrate the importance of considering the development of mating preferences to understand their within-population variation and environmentally induced change. PMID- 19812085 TI - The unexpected but understandable dynamics of mating, paternity and paternal care in the ocellated wrasse. AB - Although theory generally predicts that males should reduce paternal care in response to cues that predict increased sperm competition and decreased paternity, empirical patterns are equivocal. Some studies have found the predicted decrease in male care with increased sperm competition, while even more studies report no effect of paternity or sperm competition on male care. Here, we report the first example, to our knowledge, of paternal care increasing with the risk and intensity of sperm competition, in the ocellated wrasse (Symphodus ocellatus). Theory also predicts that if paternal care varies and is important to female fitness, female choice among males and male indicators traits of expected paternal care should evolve. Despite a non-random distribution of mating success among nests, we found no evidence for female choice among parental males. Finally, we document the highest published levels of extra-pair paternity for a species with exclusive and obligate male care: genetic paternity analyses revealed cuckoldry at 100 per cent of nests and 28 per cent of all offspring were not sired by the male caring for them. While not predicted by any existing theory, these unexpected reproductive patterns become understandable if we consider how male and female mating and parental care interact simultaneously in this and probably many other species. PMID- 19812086 TI - Evidence that two main bottleneck events shaped modern human genetic diversity. AB - There is a strong consensus that modern humans originated in Africa and moved out to colonize the world approximately 50 000 years ago. During the process of expansion, variability was lost, creating a linear gradient of decreasing diversity with increasing distance from Africa. However, the exact way in which this loss occurred remains somewhat unclear: did it involve one, a few or a continuous series of population bottlenecks? We addressed this by analysing a large published dataset of 783 microsatellite loci genotyped in 53 worldwide populations, using the program 'Bottleneck'. Immediately following a sharp population decline, rare alleles are lost faster than heterozygosity, creating a transient excess of heterozygosity relative to allele number, a feature that is used by Bottleneck to infer historical events. We find evidence of two primary events, one 'out of Africa' and one placed around the Bering Strait, where an ancient land bridge allowed passage into the Americas. These findings agree well with the regions of the world where the largest founder events might have been expected, but contrast with the apparently smooth gradient of variability that is revealed when current heterozygosity is plotted against distance from Africa. PMID- 19812087 TI - Antimicrobial chemicals in hoopoe preen secretions are produced by symbiotic bacteria. AB - Animals frequently use metabolites produced by symbiotic bacteria as agents against pathogens and parasites. Secretions from the preen gland of birds are used for this purpose, although its chemicals apparently are produced by the birds themselves. European hoopoes Upupa epops and green woodhoopoes Phoeniculus purpureus harbour symbiotic bacteria in the uropygial gland that might be partly responsible for the chemical composition of secretions. Here we investigate the antimicrobial activity of the volatile fraction of chemicals in hoopoe preen secretions, and, by means of experimental antibiotic injections, test whether symbiotic bacteria living within the uropygial gland are responsible for their production. Hoopoes produce two different kinds of secretions that differ drastically in their chemical composition. While the malodorous dark secretions produced by nestlings included a complex mix of volatiles, these chemicals did not appear in white secretions produced by non-nesting birds. All volatiles detected showed strong antibacterial activity, and a mixture of the chemicals at the concentrations measured in nestling glands inhibited the growth of all bacterial strains assayed. We found support for the hypothesized role of bacteria in the production of such antimicrobial chemicals because experimental clearance of bacteria from glands of nestlings with antibiotics resulted in secretions without most of the volatiles detected in control individuals. Thus, the presence of symbiotic bacteria in the uropygial gland provides hoopoes with potent antimicrobials for topical use. PMID- 19812088 TI - A kinetic model that explains the effect of inorganic phosphate on the mechanics and energetics of isometric contraction of fast skeletal muscle. AB - A conventional five-step chemo-mechanical cycle of the myosin-actin ATPase reaction, which implies myosin detachment from actin upon release of hydrolysis products (ADP and phosphate, Pi) and binding of a new ATP molecule, is able to fit the [Pi] dependence of the force and number of myosin motors during isometric contraction of skeletal muscle. However, this scheme is not able to explain why the isometric ATPase rate of fast skeletal muscle is decreased by an increase in [Pi] much less than the number of motors. The question can be solved assuming the presence of a branch in the cycle: in isometric contraction, when the force generation process by the myosin motor is biased at the start of the working stroke, the motor can detach at an early stage of the ATPase cycle, with Pi still bound to its catalytic site, and then rapidly release the hydrolysis products and bind another ATP. In this way, the model predicts that in fast skeletal muscle the energetic cost of isometric contraction increases with [Pi]. The large dissociation constant of the product release in the branched pathway allows the isometric myosin-actin reaction to fit the equilibrium constant of the ATPase. PMID- 19812089 TI - Toxicants and human sperm chromatin integrity. AB - The integrity of the paternal genome is essential as the spermatozoon can bring genetic damage into the oocyte at fertilization and contribute to the development of abnormal pregnancy outcome. During the past two decades, many assays have been developed to measure sperm DNA strand breaks, chromatin structure and compaction and assess the proteins associated with the DNA, as well as epigenetic modifications. Using these assays, it has been shown that exposure to physical agents or chemicals, including therapeutic drugs and environmental toxicants, can affect the integrity of sperm chromatin, inducing structural, genetic and/or epigenetic abnormalities. The mechanisms by which such damage is triggered are still largely unresolved and the susceptibility of each individual will depend on their genetic background, lifestyle and exposure to various insults. Depending on the nature of the chemicals, they may directly target the DNA, induce an oxidative stress, or modify the epigenetic elements. The significance of measuring the sperm chromatin integrity comes from the fact that this end-point correlates well with the low IVF and ICSI outcomes, and idiopathic infertility. Nevertheless, it is hard to establish a direct link between the paternal sperm chromatin integrity and the health of the future generations. Thus, it seems essential to undertake studies that will resolve the impact of chemical and environmental factors on chromatin structure and epigenetic components of human spermatozoa and to elucidate what sperm nuclear end-points are predictors of the quality of progeny outcome. PMID- 19812090 TI - SQUIRE after one year. PMID- 19812091 TI - The role of an ethical culture for the prevention and recovery of "second victims". PMID- 19812092 TI - The natural history of recovery for the healthcare provider "second victim" after adverse patient events. AB - BACKGROUND: When patients experience unexpected events, some health professionals become "second victims". These care givers feel as though they have failed the patient, second guessing clinical skills, knowledge base and career choice. Although some information exists, a complete understanding of this phenomenon is essential to design and test supportive interventions that achieve a healthy recovery. METHODS: The purpose of this article is to report interview findings with 31 second victims. After institutional review board approval, second victim volunteers representing different professional groups were solicited for private, hour-long interviews. The semistructured interview covered demographics, participant recount of event, symptoms experienced and recommendations for improving institutional support. After interviews, transcripts were analyzed independently for themes, followed by group deliberation and reflective use with current victims. RESULTS: Participants experienced various symptoms that did not differ by sex or professional group. Our analysis identified six stages that delineate the natural history of the second victim phenomenon. These are (1) chaos and accident response, (2) intrusive reflections, (3) restoring personal integrity, (4) enduring the inquisition, (5) obtaining emotional first aid and (6) moving on. We defined the characteristics and typical questions second victims are desperate to have answered during these stages. Several reported that involvement in improvement work or patient safety advocacy helped them to once again enjoy their work. CONCLUSIONS: We now believe the post-event trajectory is largely predictable. Institutional programs could be developed to successfully screen at-risk professionals immediately after an event, and appropriate support could be deployed to expedite recovery and mitigate adverse career outcomes. PMID- 19812093 TI - Enhancing medication use safety: benefits of learning from your peers. AB - BACKGROUND: Maryland hospitals have been improving the safety of medication use practices since 2000. A retrospective analysis of 35 hospitals was conducted for 2005-2007 to determine the changes in medication use practices, communication methods within hospitals, patient education and changes in medical record management. METHODS: Thirty-five Maryland hospitals completed the Institute for Safe Medication Practices Medication Safety Self-Assessment for Hospitals, a voluntary initiative to improve the safety of medication use. A weighting structure is applied to calculate key element scores, core characteristic scores and overall self-assessment scores that were used in ANOVA and regression analyses. FINDINGS: The state-wide aggregate score significantly increased from 74.2% in 2005 to 81.2% in 2007 (p<0.05). The 35 hospitals scored highest in the following key areas in 2007: drug standardisation, storage and distribution (90.2%); drug labelling, packaging and nomenclature (88.1%); and environmental factors (84.3%). Results indicated that hospitals scored lowest in the key element area related to accessibility of patient information (72.5%) and in the core characteristics pertaining to redundancies and independent double checks (64.2%) in 2007. A substantial number of hospitals had positive and significant (p<0.05) changes in certain key elements and/or core characteristics. Few hospitals showed significant (p<0.05) decreases in their scores. CONCLUSION: MEDSAFE has directly assisted Maryland hospitals in improving medication use safety. The strategies and tools of MEDSAFE have been used in Maryland since 2000 and Singapore and Austria since 2006. PMID- 19812094 TI - Estimates of injury risks for healthcare personnel working night shifts and long hours. AB - BACKGROUND: Evidence suggests that working long hours or unconventional shifts (night, evening and rotating shifts) can induce fatigue and stress in healthcare employees that might jeopardise quality of care and patient safety. METHODS: This study is based on a retrospective analysis of 13 years of occupational data from the National Longitudinal Survey of Youth, covering nearly 11,000 American workers. During the study period, 545 injuries were reported by employees in healthcare professions. Cox proportional hazard analyses were used to calculate adjusted hazard ratios comparing the risk of a job-related injury among healthcare workers in various types of demanding schedules to employees working conventional schedules. The analyses were stratified to estimate risks within different occupational classifications and care settings. RESULTS: The greatest injury risks to healthcare workers were in schedules involving overtime or at least 60 h per week. Interestingly, an elevated risk of injury was not observed for healthcare employees working 12 or more hours per day or for those in night, evening or rotating shifts. Among employees working overtime and long-hour (>60 h per week) schedules, those at medical provider offices had a significantly higher risk of injury (HR 2.86) than at hospitals, rehabilitation clinics or long-term care facilities. Support personnel, including aids, attendants, technicians, therapists and dieticians, faced a higher risk of injury than did physicians and nurses. CONCLUSION: Healthcare managers responsible for quality improvement and patient safety programmes should be aware of the possibility for worker fatigue and injury in particular scheduling arrangements. PMID- 19812095 TI - Care homes' use of medicines study: prevalence, causes and potential harm of medication errors in care homes for older people. AB - INTRODUCTION: Care home residents are at particular risk from medication errors, and our objective was to determine the prevalence and potential harm of prescribing, monitoring, dispensing and administration errors in UK care homes, and to identify their causes. METHODS: A prospective study of a random sample of residents within a purposive sample of homes in three areas. Errors were identified by patient interview, note review, observation of practice and examination of dispensed items. Causes were understood by observation and from theoretically framed interviews with home staff, doctors and pharmacists. Potential harm from errors was assessed by expert judgement. RESULTS: The 256 residents recruited in 55 homes were taking a mean of 8.0 medicines. One hundred and seventy-eight (69.5%) of residents had one or more errors. The mean number per resident was 1.9 errors. The mean potential harm from prescribing, monitoring, administration and dispensing errors was 2.6, 3.7, 2.1 and 2.0 (0 = no harm, 10 = death), respectively. Contributing factors from the 89 interviews included doctors who were not accessible, did not know the residents and lacked information in homes when prescribing; home staff's high workload, lack of medicines training and drug round interruptions; lack of team work among home, practice and pharmacy; inefficient ordering systems; inaccurate medicine records and prevalence of verbal communication; and difficult to fill (and check) medication administration systems. CONCLUSIONS: That two thirds of residents were exposed to one or more medication errors is of concern. The will to improve exists, but there is a lack of overall responsibility. Action is required from all concerned. PMID- 19812096 TI - Do associations between staff and inpatient feedback have the potential for improving patient experience? An analysis of surveys in NHS acute trusts in England. AB - OBJECTIVE: To examine whether staff feedback on quality, safety and workforce issues is reflected in patient-reported experience. SETTING: 166 NHS acute trusts in England. PARTICIPANTS: Respondents to the 2006 surveys of adult inpatients and staff in 166 NHS acute trusts in England. METHODS: Multiple linear regression was used to model the relationship between responses by "frontline" staff and inpatients at trust level. Staff survey items were the explanatory variables and inpatient responses the dependent variables. Adjustments were made for location (London and non-London) and trust type. RESULTS: 69 500 staff and 81 000 patients responded to the surveys. There were several significant associations between staff and patients' responses, including that staff availability of hand-washing materials was positively associated with patient feedback on cleanliness and hand washing by doctors/nurses (p<0.00). It was a significant predictor of patient experience also in several other models. Other significant predictors of patient experience were managerial support, witnessing and reporting of errors (positively associated with patient experience), working extra hours and stress (negatively associated). London trusts performed worse on patient experience than trusts outside London and specialist trusts performed better than other acute trusts. CONCLUSIONS: Staff feedback was associated with patient-reported experience. Positive staff feedback on availability of hand-washing materials was broadly reflective of positive patient experience. Negative staff experience was reflected in poorer patient experience and vice versa. Although we cannot demonstrate causality, the consistent direction of the findings is indicative of it. Management boards of trusts and clinicians and other staff should monitor and act on the results of their staff surveys. This has the potential for improving quality, safety and patient experience. PMID- 19812097 TI - Improving colon cancer screening rates in primary care: a pilot study emphasising the role of the medical assistant. AB - BACKGROUND: Colorectal cancer (CRC) is the third-leading cause of cancer death for both men and women in the USA. Despite consensus recommendations for screening, just over half of eligible adults nationally have undergone screening. We therefore implemented a programme to improve the rate of CRC screening. METHODS: This study was conducted in the Utah Health Research Network and the University of Utah Community Clinics, a 100 000 patient, seven-practice, university-owned system offering primary and secondary care and ancillary services including endoscopy. We focused on patients aged >or=50 who were seen between 1 January 2003 and 31 October 2006, and who were not current for CRC screening at the time of the visit. The study included a three-phase INTERVENTION: electronic medical record (EMR) reminders, physician and medical assistant (MA) education about CRC screening guidelines, and redesign of patient visit workflow with an expanded role for MAs to review patients' CRC screening status and recommend testing when appropriate. With patient agreement, the MA entered a preliminary order in the EMR, and the physician confirmed or rejected the order. The primary outcome measure was the rate of screening colonoscopy ordered for eligible patients. RESULTS: The baseline colonoscopy referral rate was 6.0%. Provider education and electronic reminders had minimal immediate impact on screening rates. Addition of the expanded MA role was associated with a sustained increase in colonoscopy referral order rate to 13.4%, a relative improvement of 123%. CONCLUSIONS: The MA can play a key role in improving CRC screening rates as part of a redesigned system of primary care. PMID- 19812098 TI - ProvenCare: quality improvement model for designing highly reliable care in cardiac surgery. AB - OBJECTIVE: To test whether an integrated delivery system could, through the application of process redesign methodology and reliability science, implement multiple evidence-based medical practices across the continuum of care for a specific surgical intervention and deliver these practices consistently. METHODS: The programme-ProvenCare--had three components: establishing best practices for elective coronary artery bypass graft (CABG) patients; assembling a multidisciplinary team to "hardwire" these best practices into everyday workflow; and implementing the programme with real-time data collection, feedback and focused redesign to reach high reliability. Surgeons reviewed all class I and IIa 2004 ACC/AHA guidelines for CABG surgery and translated them into 19 clinically applicable recommendations. A frontline multidisciplinary team "hardwired" these, resulting in 40 measurable process elements. Feedback of gaps in care was given and the process redesigned as needed. Clinical outcome data on consecutive elective CABG patients seen in the 12 months pre-intervention were then compared with a post-intervention group. RESULTS: Initially, 59% of patients received all 40 elements. At 3 months, compliance reached 100%, fell transiently to 86% and then reached 100% again, and was sustained for the remainder of the study. The overall trend in reliability was significant (p = 0.001). 30-day clinical outcomes showed improved trends in 8/9 measured areas (eg, patient readmissions to ICU decreased from 2.9% to 0.9% and blood products usage decreased from 23.4% to 16.2%). Operative mortality decreased to zero, but only likelihood of discharge was significant (p = 0.033). Frequency and length of readmissions fell, as did mean hospital charges. CONCLUSION: Frontline medical care providers, led by process design specialists, can successfully redesign episodic processes to consistently deliver evidence-based medicine, which may improve patient outcomes and reduce resource use. PMID- 19812099 TI - Reduced in-hospital mortality for heart failure with clinical pathways: the results of a cluster randomised controlled trial. AB - BACKGROUND: Hospital treatment of heart failure (HF) frequently does not follow published guidelines, potentially contributing to HF high morbidity, mortality and economic cost. The Experimental Prospective Study on the Effectiveness and Efficiency of the Implementation of Clinical Pathways was undertaken to determine how clinical pathways (CP) for hospital treatment of HF affected care variability, guidelines adherence, in-hospital mortality and outcomes at discharge. Methods/ DESIGN: Two-arm, cluster-randomised trial. Fourteen community hospitals were randomised either to the experimental arm (CP: appropriate therapeutic guidelines use, new organisation and procedures, patient education) or to the control arm (usual care). The main outcome was in-hospital mortality; secondary outcomes were length and appropriateness of the stay, rate of unscheduled readmissions, customer satisfaction, usage of diagnostic and therapeutic procedures during hospital stay and quality indicators at discharge. All outcomes were measured using validated instruments available in literature. RESULTS: In-hospital mortality was 5.6% in the experimental arm (n = 12); 15.4% in controls (n = 33, p = 0.001). In CP and usual care groups, the mean rates of unscheduled readmissions were 7.9% and 13.9%, respectively. Adjusting for age, smoking, New York Heart Association score, hypertension and source of referral, patients in the CP group, as compared to controls, had a significantly lower risk of in-hospital death (OR 0.18; 95% CI 0.07 to 0.46) and unscheduled readmissions (OR 0.42; 95% CI 0.20 to 0.87). No differences were found between CP and control with respect to the appropriateness of the stay, costs and patient's satisfaction. Except for electrocardiography, all recommended diagnostic procedures were used more in the CP group. Similarly, pharmaceuticals use was significantly greater in CP, with the exception of diuretics and anti-platelets agents. DISCUSSION: The introduction of a specifically tailored CP for the hospital treatment of HF was effective in reducing in-hospital mortality and unscheduled readmissions. This study adds to previous knowledge indicating that CP should be used to improve the quality of hospital treatment of HF. TRIAL REGISTRATION NUMBER: NCT00519038. PMID- 19812100 TI - Assessment of patients' tendency to give a positive or negative rating to healthcare. AB - BACKGROUND: Adjustment of patient satisfaction scores for case-mix variables such as age and sex may lead to overadjustment. The patient's tendency to rate healthcare positively or negatively may be the only variable that should be adjusted to improve the comparability of satisfaction scores between healthcare providers. OBJECTIVE: To develop a measure of "rating tendency", assess its stability over time, explore its distribution across subgroups of patients and its association with patient opinion scores. DESIGN AND SUBJECTS: A scale based on 10 hypothetical scenarios describing hospital care episodes was developed. It was administered both before and after hospitalisation to 203 patients programmed for elective orthopaedic surgery in a Swiss teaching hospital. A problem score regarding the actual hospitalisation was obtained at follow-up. RESULTS: The rating tendency scale had good internal consistency (Cronbach alpha 0.85), and factor analysis confirmed that it measured a single underlying concept. However, the correlation between prehospitalisation and posthospitalisation measures was moderate (intraclass correlation coefficient 0.55, p<0.001), as was the correlation with the hospitalisation problem score (Spearman r = -0.22, p = 0.002). The pattern of rating tendency across subgroups of respondents mirrored the pattern of problem scores. Adjusting for the rating tendency had little effect on comparisons of problem scores between subgroups of patients, all of whom were treated at the same hospital. CONCLUSIONS: A patient's "rating tendency" can be measured using a reliable 10-item scale. The utility of adjusting satisfaction scores for rating tendency when comparing hospitals remains to be tested. PMID- 19812101 TI - Safety analysis of patient transfers and handling tasks. AB - BACKGROUND: Low-back disorders are related to biomechanical demands, and nurses are among the professionals with the highest rates. Quantification of risk factors is important for safety assessment and reduction of low-back disorders. OBJECTIVE: This study aimed to quantify physical demands of frequent nursing tasks and provide evidence-based recommendations to increase low-back safety. METHODS: Thirty-six volunteer female nurses participated in a cross-sectional study of nine nursing tasks. Lumbar range of motion (ROM) and motion during nursing tasks were measured. Compression and shear forces at L5/S1, ligament strain and percentage of population without sufficient torso strength to perform 14 phases of nine nursing tasks were estimated. RESULTS: Peak flexions during trolley-to-bed, bed-to-chair and chair-to-bed transfers reached the maximum flexion ROM of the nurses. Average lumbar flexion during trolley-to-bed transfers was >50% of flexion ROM, being higher than during all other tasks. Mean (SD) compression at L5/S1 (4754 N (437 N)) and population without sufficient torso strength (37% (9%)) were highest during the pushing phase of bed-to-trolley transfers. Shear force (487 N (40 N)) and ligament strain (14% (5%)) were highest during the pulling phase of trolley-to-bed transfers. CONCLUSIONS: Nursing tasks impose high biomechanical demands on the lumbar spine. Excessive lumbar flexion and forces are critical aspects of manual transfers requiring most of the nurses' capabilities. Evidence-based recommendations to improve low-back safety in common nursing tasks were provided. Fitness to work, job modifications and training programs can now be designed and assessed based on the results. PMID- 19812102 TI - Effects of evidence-based clinical practice guidelines on quality of care: a systematic review. AB - BACKGROUND: Evidence-based clinical guidelines aim to improve the quality of care. In The Netherlands, considerable time and effort have been invested in the development and implementation of evidence-based guidelines since the 1990s. Thus far, no reviews are available on their effectiveness. The primary aim of this article was to assess the evidence for the effectiveness of Dutch evidence-based clinical guidelines in improving the quality of care. METHODS: A systematic review of studies evaluating the effects of Dutch evidence-based guidelines on both the process and structure of care and patient outcomes was conducted. The electronic databases Medline and Embase (1990-2007) and relevant scientific journals were searched. Studies were only selected if they included a controlled trial, an interrupted time series design or a before and after design. RESULTS: A total of 20 studies were included. In 17 of 19 studies that measured the effects on the process or structure of care, significant improvements were reported. Thirteen of these studies reported improvement with respect to some of the recommendations studied. In addition, the size of the observed effects varied largely across the recommendations within guidelines. Six of nine studies that measured patient health outcomes showed significant but small improvements as a result of the use of clinical guidelines. CONCLUSIONS: This review demonstrates that Dutch evidence-based clinical guidelines can be effective in improving the process and structure of care. The effects of guidelines on patient health outcomes were studied far less and data are less convincing. The high level of variation in effects across recommendations suggests that implementation strategies tailored to individual recommendations within the guideline are needed to establish relevant improvements in healthcare. Moreover, the results highlight the need for well-designed studies focusing on the level of the recommendations to determine which factors influence guideline utilisation and improved patient outcomes. PMID- 19812103 TI - Development and usability of a behavioural marking system for performance assessment of obstetrical teams. AB - BACKGROUND: Teamwork and communication have been identified as root causes of sentinel events involving infant death and injury during delivery. However, despite the emphasis on team training as a way to improve maternal and fetal safety outcomes, valid and reliable markers of obstetrical team performance are not available to assess curricular efficacy. OBJECTIVES: The objective of this study was to develop and assess the usability of two obstetrical behavioural marking systems for use with simulation entitled Assessment of Obstetrical Team Performance (AOTP) and Global Assessment of Obstetrical Team Performance (GAOTP). METHODS: In a previous study, obstetrical teams were videotaped managing simulated emergency obstetrical scenarios. In the current study, 13 reviewers reviewed these videotapes and generated a list of behaviours judged to negatively or positively affect the teams' performances. Qualitative analysis using research team consensus and NVivo generated themes and subthemes. Research team members developed descriptors for poor and excellent team performance for each of the behaviours. Subsequently, the usability of the prototypes was assessed by an additional 14 reviewers. RESULTS: In total, the reviewers identified 1294 items, which were sorted into 6 themes and 18 subthemes of obstetrical team performance. In terms of usability, the median amount of time that participants spent completing the AOTP was 7.5 min (range 1.5 to 50 min) and 75% thought the time requirement was moderate and manageable. CONCLUSION: Feedback regarding usability suggests that the AOTP allows for an accurate reflection of raters' assessments of the performance of the team, and as a whole, it is comprehensive, quick and easy to use. Studies are underway to establish the validity and reliability of the AOTP and GAOTP. PMID- 19812104 TI - Exploring policy makers' perspectives on a clinical controversy: airway surgery for adult obstructive sleep apnoea. AB - BACKGROUND AND OBJECTIVES: Worldwide, there is increasing focus on measures to reduce ineffective healthcare practices. Upper airway surgeries for the treatment of adult obstructive sleep apnoea (OSA) represent a case-study in this area, given recent publications that draw into question their efficacy. Policy stakeholders were canvassed to assess their perspectives on this. DESIGN AND SETTING: Senior health policy stakeholders from Australia were criterion and snowball sampled (to identify opinion leaders). Participants were presented with preparatory material and took part in individual semistructured interviews. These focused on eliciting responses to recently published evidence and a relevant Cochrane review. Questions were posed relating to clinical effectiveness and associated policy implications. Interviews were taped and transcribed for thematic analysis. Participant comments were de-identified. FINDINGS: Ten stakeholders were interviewed before saturation was reached. Thematic analysis highlighted participant concern with the diversity of procedures on offer, coupled with limited effectiveness (suggesting potential clinical uncertainty) and considerations therefore of resource allocation (potential opportunity cost). Stakeholders seem aware of the methodological complexities, the ethical issues raised and the role of patients in considerations regarding appropriateness. Finally, policy stakeholders acknowledge that these procedures appear appropriate only for a minority, with consensus that policy level restrictions to government funding for these procedures may be warranted. CONCLUSION: This report highlights that this clinical controversy is of interest and relevance from a policy perspective with lessons and potential implications for clinical practice. It further highlights the need for clinical consensus on definitions of surgical "success" in treating this condition, as this forms an important pretext to policy considerations. PMID- 19812105 TI - Medication reconciliation in ambulatory care: attempts at improvement. AB - OBJECTIVE: To enhance overall accuracy of medication lists by providing performance feedback and training to the healthcare team and increasing patient participation in the medication reconciliation process. METHODS: This prospective study involved patients seen in four academic, ambulatory primary care internal medicine clinics. Before the interventions, baseline data were analysed, assessing completeness, correctness and accuracy of medication documentation in the electronic medical record. Interventions to provide performance feedback and training to the healthcare team, increase patient awareness and participation in the medication reconciliation process were implemented. Immediately after each intervention, a data collection was undertaken to assess the effectiveness of the intervention on the accuracy of individual medications and medication lists. RESULTS: Completeness of medication lists improved from 20.4% to 50.4% (p<0.001). The incomplete documentation of medication lists was mostly because of lack of frequency (15.4%) and route (8.9%) for individual medications within a medication list. Correctness of medication lists improved from 23.1% to 37.7% (p = 0.087). The incorrectness in a medication list was mostly because of incorrect medications dose. Patient participation in the medication reconciliation process increased from 13.9% to 33% (p<0.001). The medication list accuracy improved from 11.5% to 29% (p = 0.014). CONCLUSION: In this setting, it was helpful to engage the active participation of all members of the healthcare team and most importantly the patient to improve the accuracy of medication lists. PMID- 19812106 TI - Improving the use of venous thromboembolism prophylaxis in an Australian teaching hospital. AB - INTRODUCTION: Hospital-acquired venous thromboembolism (VTE) is a potentially fatal complication of hospitalisation, with meta-analyses and guidelines supporting the use of proven prophylactic measures (graduated compression stockings (GCS) and anticoagulants). Despite this, prophylaxis is underutilised and represents one of the largest gaps between evidence and current clinical practice. METHODS: All episodes of VTE complicating hospitalisation were ascertained prospectively as part of a quality improvement programme over 3.5 years with a view to designing interventions to improve the use of prophylaxis and reduce the rate of VTE. Interventions initially centred upon highlighting the burden of VTE, the extent of failure to apply guideline evidence into practice, and the development and application of a hospital-wide risk assessment tool. Later interventions sought to build the risk-assessment tool into routine clinical care and enhanced feedback on VTE to clinical teams. RESULTS: The annual rate of VTE fell in all the years following the intervention (2001), from 2.57 per 1000 cost-weighted separations to a nadir of 1.87 in 2003, with the difference being statistically significant (RR 0.68, 0.47 to 0.99, p = 0.04). The proportion of patients receiving anticoagulant prophylaxis increased (48% to 74%, p = 0.01) but there was no change in the measured use of GCS. There was a marked increase in the use of risk assessment for VTE in the ward setting (7.7% to 100%, p<0.001) during the programme. CONCLUSION: Affordable and accessible interventions can improve the application of VTE prophylaxis guidelines into daily hospital care and are associated with reductions in this potentially life threatening complication. PMID- 19812107 TI - Frank and Lillian Gilbreth: scientific management in the operating room. PMID- 19812109 TI - Genome structure of a Saccharomyces cerevisiae strain widely used in bioethanol production. AB - Bioethanol is a biofuel produced mainly from the fermentation of carbohydrates derived from agricultural feedstocks by the yeast Saccharomyces cerevisiae. One of the most widely adopted strains is PE-2, a heterothallic diploid naturally adapted to the sugar cane fermentation process used in Brazil. Here we report the molecular genetic analysis of a PE-2 derived diploid (JAY270), and the complete genome sequence of a haploid derivative (JAY291). The JAY270 genome is highly heterozygous (approximately 2 SNPs/kb) and has several structural polymorphisms between homologous chromosomes. These chromosomal rearrangements are confined to the peripheral regions of the chromosomes, with breakpoints within repetitive DNA sequences. Despite its complex karyotype, this diploid, when sporulated, had a high frequency of viable spores. Hybrid diploids formed by outcrossing with the laboratory strain S288c also displayed good spore viability. Thus, the rearrangements that exist near the ends of chromosomes do not impair meiosis, as they do not span regions that contain essential genes. This observation is consistent with a model in which the peripheral regions of chromosomes represent plastic domains of the genome that are free to recombine ectopically and experiment with alternative structures. We also explored features of the JAY270 and JAY291 genomes that help explain their high adaptation to industrial environments, exhibiting desirable phenotypes such as high ethanol and cell mass production and high temperature and oxidative stress tolerance. The genomic manipulation of such strains could enable the creation of a new generation of industrial organisms, ideally suited for use as delivery vehicles for future bioenergy technologies. PMID- 19812110 TI - Knowledge-based instantiation of full atomic detail into coarse-grain RNA 3D structural models. AB - MOTIVATION: The recent development of methods for modeling RNA 3D structures using coarse-grain approaches creates a need to bridge low- and high-resolution modeling methods. Although they contain topological information, coarse-grain models lack atomic detail, which limits their utility for some applications. RESULTS: We have developed a method for adding full atomic detail to coarse-grain models of RNA 3D structures. Our method [Coarse to Atomic (C2A)] uses geometries observed in known RNA crystal structures. Our method rebuilds full atomic detail from ideal coarse-grain backbones taken from crystal structures to within 1.87 3.31 A RMSD of the full atomic crystal structure. When starting from coarse-grain models generated by the modeling tool NAST, our method builds full atomic structures that are within 1.00 A RMSD of the starting structure. The resulting full atomic structures can be used as starting points for higher resolution modeling, thus bridging high- and low-resolution approaches to modeling RNA 3D structure. AVAILABILITY: Code for the C2A method, as well as the examples discussed in this article, are freely available at www.simtk.org/home/c2a. CONTACT: russ.altman@stanford.edu PMID- 19812111 TI - Histone deacetylase-2 and airway disease. AB - The increased expression of inflammatory genes in inflammatory lung diseases is regulated by acetylation of core histones, whereas histone deacetylase-2 (HDAC2) suppresses inflammatory gene expression. Corticosteroids suppress inflammatory genes in asthma by inhibiting histone acetyltransferase and in particular by recruiting HDAC2 to the nuclear factor-kappaB-activated inflammatory gene complex. This involves deacetylation of the acetylated glucocorticoid receptor. In COPD, severe asthma and asthmatics who smoke, HDAC2 is reduced, thus preventing corticosteroids from suppressing inflammation. The reduction in HDAC2 appears to be secondary to increased oxidative and nitrative stress in the lungs. Antioxidants and inhibitors of nitric oxide synthesis may therefore restore corticosteroid sensitivity in COPD, but this can also be achieved by low concentrations of theophylline and curcumin, which act as HDAC activators. Theophylline is a direct inhibitor of oxidant-activated phosphoinositide-3-kinase delta, which is involved in inactivation of HDAC2. In the future selective PI3Kdelta inhibitors and more direct activators of HDAC2 may be used to treat corticosteroid-resistant inflammatory diseases of the lung, including COPD, severe asthma and asthma in smokers. PMID- 19812114 TI - Excess mortality and cause of death in a cohort of Norwegian multiple sclerosis patients. AB - There are few studies of long-term, cause-specific mortality in multiple sclerosis (MS) relating to population mortality. Our objective was to study survival, excess mortality and causes of death in a cohort of patients with a long history of MS. Patients living in Oslo with definite MS and onset during 1940-80 were included in 2006. Causes of death and mortality in the general population were obtained from the Cause of Death Registry of Statistics Norway. Of the 386 patients included in the study, 263 (68%) had died at inclusion. Median survival from onset was 35 years (Kaplan-Meier: 95% confidence interval 33 37). Primary progressive MS was associated with shorter survival, but mean age at death was similar for relapsing-remitting and primary progressive MS patients. The most frequent underlying cause of death was MS (50%), and infection was often registered as a contributory cause (56%). The all-cause standardized mortality ratio was 2.47. Excess mortality was most marked during the second decade after onset of MS. We conclude that infections are probably the main cause of death in patients with MS, but the frequency is underestimated due to misleading information on death certificates. Excess mortality in patients with MS first appeared during the second decade of the disease. Survival seems to be age dependent rather than related to disease course. PMID- 19812115 TI - Hypothalamic stimulation for trigeminal neuralgia in multiple sclerosis patients: efficacy on the paroxysmal ophthalmic pain. AB - Trigeminal neuralgia is a disorder characterized by paroxysmal pain arising in one or more trigeminal branches; it is commonly reported in multiple sclerosis. In multiple sclerosis patients the ophthalmic branch may be frequently involved and the risks carried by neurosurgical ablative procedures are higher including major adverse effects such as corneal reflex impairment and keratitis. The objective of this works is to assess the role of posterior hypothalamus neuromodulation in the treatment of trigeminal neuralgia in multiple sclerosis patients. Five multiple sclerosis patients suffering from refractory recurrent trigeminal neuralgia involving all three trigeminal branches underwent deep brain stimulation of the posterior hypothalamus. The rationale of this intervention emerges from our earlier success in treating pain patients suffering from trigeminal autonomic cephalalgias. After follow-up periods that ranged from 1 to 4 years after treatment, the paroxysmal pain arising from the first trigeminal branch was controlled, whereas the recurrence of pain in the second and third trigeminal branches necessitated repeated thermorhizotomies to control in pain in two patients after 2 years of follow-up. In conclusion, deep brain stimulation may be considered as an adjunctive procedure for treating refractory paroxysmal pain within the first trigeminal division so as to avoid the complication of corneal reflex impairment that is known to follow ablative procedures. PMID- 19812116 TI - Severe relapses after the first infusion of natalizumab in active relapsing remitting multiple sclerosis. AB - We describe three patients suffering from a very active form of relapsing remitting multiple sclerosis (MS), who experienced severe disease worsening, associated with a marked increase in brain inflammation, a few days after the first administration of natalizumab. In line with preclinical studies, our observations suggest that natalizumab, when administered during active disease phases, may worsen disease evolution possibly by modifying the regulatory network in the brain. We suggest that relapsing-remitting MS patients having had a recent relapse should be treated with natalizumab only after achieving complete clinical and radiological remission. PMID- 19812117 TI - Clinically isolated acute transverse myelitis: prognostic features and incidence. AB - Demyelinating acute transverse myelitis may be the first presentation of multiple sclerosis or remain a clinically isolated syndrome. North Canterbury, New Zealand provides a well circumscribed population to study acute transverse myelitis. OBJECTIVE: to identify prognostic features, clinical outcomes and incidence of ATM in North Canterbury, New Zealand. All patients with acute transverse myelitis as a first neurological presentation diagnosed from January 2001 to December 2005 at a single institution providing all neurological care for North Canterbury were assessed for clinical data, MRI findings, cerebrospinal fluid results and clinical outcomes. CHAMPS, Barkhof/Tintore and Swanton criteria were applied to brain MRI. Sixty-one patients were identified with a mean duration of follow-up of 30 +/- 17 months. Fifty percent of patients with ATM with brain lesions by CHAMPS criteria converted to clinically definite multiple sclerosis. No patients with idiopathic acute transverse myelitis converted to clinically definite multiple sclerosis. There was a strong association with conversion to clinically definite multiple sclerosis and abnormal brain MRI by CHAMPS criteria (hazard ratio, 5.63; 1.83-17.3), Barkhof/Tintore criteria (hazard ratio, 6.43; 2.31-17.9) and Swanton criteria (hazard ratio, 4.53; 1.67-12.3). The age standardized annual incidence of acute transverse myelitis was 24.6 (18.2-31.1) per million, of definite and possible idiopathic acute transverse myelitis was 6.2 (2.9-9.6) per million, and of acute transverse myelitis with brain lesions was 4.7 (1.9-7.6) per million. Patients with idiopathic acute transverse myelitis are at low risk for conversion to clinically definite multiple sclerosis. Abnormal brain MRI by CHAMPS criteria is a sensitive predictor of conversion to clinically definite multiple sclerosis. The annual incidence of acute transverse multiple sclerosis in North Canterbury, New Zealand is significantly higher than previously reported. PMID- 19812118 TI - Severe acetaminophen poisoning treated with a fractionated plasma separation and absorption system: A case report. AB - Acetaminophen is an analgesic drug that is frequently used in suicide attempts. In this paper, we report on a 17-year-old girl who was admitted to an emergency department 15 hours after taking acetaminophen pills in a suicide attempt. Her serum acetaminophen level was 73 mg/L on admission; she had elevated liver enzymes suggesting hepatic necrosis. She was started on N-acetyl cystein (NAC), and treated successfully with a fractionated plasma separation and absorption system. PMID- 19812119 TI - Attenuation of Helicteres isora L. bark extracts on streptozotocin-induced alterations in glycogen and carbohydrate metabolism in albino rats. AB - The present study was undertaken to assess the effect of Helicteres isora L. on four important enzymes of carbohydrate metabolism (glucokinase [GK], hexokinase [HK] phosphofructokinase [PFK] and fructose-1, 6-bisphosphatase [FBP]) along with glycogen content of insulin-dependent (skeletal muscle and liver) and insulin independent tissues (kidneys and brain) in streptozotocin (STZ; 60 mg/kg)-induced model of diabetes for 30 days. Administration of bark extracts (100, 200 mg/kg) for 30 days led to decrease in plasma glucose levels by approximately 9.60% and 22.04% and 19.18% and 33.93% on 15th and 30th day, respectively, of the experiment. Liver and two-kidney weight expressed as percentage of body weight significantly increased in diabetics (P < 0.05) versus normal controls. Renal glycogen content increased by 10 folds while hepatic and skeletal muscle glycogen content decreased by 75% and 68% in diabetic controls versus controls. H. isora did not affect glycogen content in any tissue. The decreased activities of PFK, GK, FBP and HK in diabetic controls were 40%, 50%, 50% and 60% and bark extract of H. isora partially corrected this alteration. The efficacy of the bark extract was comparable with Tolbutamide, a well-known hypoglycemic drug. PMID- 19812120 TI - Cardiac biomarkers in a model of acute catecholamine cardiotoxicity. AB - Coronary heart disease and in particular its most serious form - acute myocardial infarction (AMI) - represents the most common cause of mortality in developed countries. Better prognosis may be achieved by understanding the etiopathogenetic mechanisms of AMI. Therefore, a catecholamine model of myocardial injury, which has appeared to be very similar to AMI in human in some aspect, was used. Male Wistar:Han rats were randomly divided into two groups: control group (saline) and isoprenaline group (ISO; synthetic catecholamine, 100 mg.kg(- 1) subcutaneously [s.c.]). After 24 hours, functional parameters were measured, biochemical markers in the blood and metals content in the heart tissue were analysed and histological examination was performed. ISO caused marked myocardial injury that was associated with myocardial calcium overload. Close correlation between myocardial impairment (i.e. serum TnT, stroke volume index and wet ventricles weight) and the levels of myocardial calcium was observed. Direct reactive oxygen species (ROS) involvement was documented only by non-significant increase in malonyldialdehyde 24 hours after ISO injury. Moreover, myocardial element analysis revealed no significant changes as for the content of zinc and iron while selenium and copper increased in the ISO group although it reached statistical significance only for the latter. PMID- 19812121 TI - Severe scorpion envenomation in children: Management in pediatric intensive care unit. AB - BACKGROUND: Scorpion envenomation is a common public health problem worldwide and children are at greater risk of developing severe cardiac, respiratory and neurological complications. The aim of this study was to evaluate the effects of antivenin and/or prazosin use on prognosis of scorpion-envenomed children admitted to pediatric intensive care unit (PICU). METHODS: The standardized medical records of 45 children hospitalized with severe scorpion sting in PICU were retrospectively evaluated. General characteristics of the children, clinical and laboratory findings, treatment approaches and prognosis were evaluated. RESULTS: The mean age of the patients were 6.1 +/- 4.1 years ranging between 4 month and 15 years. Male to female ratio was 1.8. Thirty-three (71.1%) cases of scorpion stings came from rural areas. Twenty-six (57.8%) of the patients were stung by Androctonus crassicauda. The most common sting localization was the foot leg (55.6%). The mean duration from the scorpion sting to hospital admission was 4.5 +/- 2.6 hours. The most common findings at presentation were cold extremities (95.5%), excessive sweating (91.1%) and tachycardia (77.7%). The mean leukocyte count, and serum levels of glucose, lactate dehydrogenase, creatine phosphokinase and international normalized ratio were found above the normal ranges. Prazosin was used in all patients, dopamine in 11 (24.4%) and Na-nitroprusside in 4 (8.8%) patients. Two children died (4.4%) due to pulmonary oedema. These children, in poor clinical status at hospital admission, needed mechanical ventilation, and death occurred despite use of antivenin and prazosin in both of them. CONCLUSION: The current management of children with severe scorpion envenomation consists of administration of specific antivenom and close surveillance in a PICU, where vital signs and continuous monitoring enable early initiation of therapy for life threatening complications. The aggressive medical management directed at the organ system specifically can be effective. Our data indicated that when admission to hospital is late, the beneficial effect of antivenom and/or prazosin is questionable in severe scorpion stings. PMID- 19812122 TI - Spider bite (latrodectism) in Mashhad, Iran. AB - BACKGROUND: Spider (Latrodectus tredecimguttatus) bites are relatively common in North East Iran. They induce morbidity and rarely mortality. We aimed to investigate clinical, electrocardiographic and para-clinical changes in patients with this bite. METHODS: All consecutive patients admitted with suspected spider bites between September 2005 and September 2006 were studied prospectively. RESULTS: Spider bites accounted for 56 cases (0.5% of all poisoning, 21% of all admitted envenomated). The patients' mean (SD) age was 32 (16) years. The most common findings were pain (90%) mainly in their back (45%), stomach (35%), lower limbs (33%), upper limbs (19%) and chest (14%). Other clinical findings included were sweating (55%), chills (29%), dyspnea (25%), flushing (14%), spasm (12%), headache (12%), nausea (12%) and vertigo (12%). On electrocardiograph (ECG); ST segments were depressed in 25% of cases in at least two of the pre-cordial leads. Laboratory findings were in normal ranges. All the patients recovered following supportive and symptomatic treatment (no anti-toxin available in Iran), with a mean hospitalization period of 1.9 (1.3) days. CONCLUSIONS: Spider (L. Tredecimguttatus) bite is relatively common in Mashhad, which induces latrodectism with relatively different findings and cardiac toxicity. ECG monitoring should be considered, particularly when specific anti-toxin is not available. PMID- 19812123 TI - Acrylamide-induced oxidative stress in human erythrocytes. AB - Acrylamide (AA), a widely used industrial chemical, is shown to be neurotoxic, mutagenic and carcinogenic. This study was carried out to investigate the effects of different doses of AA on lipid peroxidation (LPO), haemolysis, methaemoglobin (MetHb) and antioxidant system in human erythrocytes in vitro. Erythrocyte solutions were incubated with 0.10, 0.25, 0.50 and 1.00 mM of AA at 37 degrees C for 1 hour. At the end of the incubation, malondialdehyde (MDA), an end product of LPO, was determined by liquid chromatography (LC) while total glutathione, reduced glutathione (GSH) levels, activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) enzymes and the rates of haemolysis and MetHb were determined by spectrophotometric methods. All of the studied concentrations of AA increased MetHb formation and SOD activity, and induced MDA formation and haemolysis due to the destruction of erythrocyte cell membrane. AA caused a decrease in the activities of GSH-Px, CAT and GSH levels. However, these effects of AA were seen only at higher concentrations than AA intake estimated for populations in many countries. We suggest that LPO process may not be involved in the toxic effects of AA in low concentrations, although the present results showed that the studied concentrations of AA exert deteriorating effects on antioxidant enzyme activities, LPO process and haemolysis. PMID- 19812124 TI - Ammonium sulphate precipitation to measure antibodies to double-stranded DNA: a simple clinical test for diagnosis and management of patients with systemic lupus erythematosus. PMID- 19812125 TI - Protein-losing enteropathy with mesenteric venous thrombosis: a forme fruste of systemic lupus erythematosus. PMID- 19812126 TI - Rapamycin impairs metabolism-secretion coupling in rat pancreatic islets by suppressing carbohydrate metabolism. AB - Rapamycin, an immunosuppressant used in human transplantation, impairs beta-cell function, but the mechanism is unclear. Chronic (24 h) exposure to rapamycin concentration dependently suppressed 16.7 mM glucose-induced insulin release from islets (1.65+/-0.06, 30 nM rapamycin versus 2.35+/-0.11 ng/islet per 30 min, control, n=30, P<0.01) without affecting insulin and DNA contents. Rapamycin also decreased alpha-ketoisocaproate-induced insulin release, suggesting reduced mitochondrial carbohydrate metabolism. ATP content in the presence of 16.7 mM glucose was significantly reduced in rapamycin-treated islets (13.42+/-0.47, rapamycin versus 16.04+/-0.46 pmol/islet, control, n=30, P<0.01). Glucose oxidation, which indicates the velocity of metabolism in the Krebs cycle, was decreased by rapamycin in the presence of 16.7 mM glucose (30.1+/-2.7, rapamycin versus 42.2+/-3.3 pmol/islet per 90 min, control, n=9, P<0.01). Immunoblotting revealed that the expression of complex I, III, IV, and V was not affected by rapamycin. Mitochondrial ATP production indicated that the respiratory chain downstream of complex II was not affected, but that carbohydrate metabolism in the Krebs cycle was reduced by rapamycin. Analysis of enzymes in the Krebs cycle revealed that activity of alpha-ketoglutarate dehydrogenase (KGDH), which catalyzes one of the slowest reactions in the Krebs cycle, was reduced by rapamycin (10.08+/-0.82, rapamycin versus 13.82+/-0.84 nmol/mg mitochondrial protein per min, control, n=5, P<0.01). Considered together, these findings indicate that rapamycin suppresses high glucose-induced insulin secretion from pancreatic islets by reducing mitochondrial ATP production through suppression of carbohydrate metabolism in the Krebs cycle, together with reduced KGDH activity. PMID- 19812127 TI - Carbon-ion radiotherapy: clinical aspects and related dosimetry. AB - The features of relativistic carbon-ion beams are attractive from the viewpoint of radiotherapy. They exhibit not only a superior physical dose distribution but also an increase in biological efficiency with depth, because energy loss of the beams increases as they penetrate the body. This paper reviews clinical aspects of carbon-beam radiotherapy using the experience at the National Institute of Radiological Sciences. The paper also outlines the dosimetry related to carbon beam radiotherapy, including absolute dosimetry of the carbon beam, neutron measurements and radiation protection measurements. PMID- 19812129 TI - Operational radiation protection in high-energy physics accelerators. AB - An overview of operational radiation protection (RP) policies and practices at high-energy electron and proton accelerators used for physics research is presented. The different radiation fields and hazards typical of these facilities are described, as well as access control and radiation control systems. The implementation of an operational RP programme is illustrated, covering area and personnel classification and monitoring, radiation surveys, radiological environmental protection, management of induced radioactivity, radiological work planning and control, management of radioactive materials and wastes, facility dismantling and decommissioning, instrumentation and training. PMID- 19812131 TI - Mobilisation or immobilisation for cervical radiculopathy? PMID- 19812130 TI - Cervical collar or physiotherapy versus wait and see policy for recent onset cervical radiculopathy: randomised trial. AB - OBJECTIVE: To evaluate the effectiveness of treatment with collar or physiotherapy compared with a wait and see policy in recent onset cervical radiculopathy. DESIGN: Randomised controlled trial. SETTING: Neurology outpatient clinics in three Dutch hospitals. PARTICIPANTS: 205 patients with symptoms and signs of cervical radiculopathy of less than one month's duration INTERVENTIONS: Treatment with a semi-hard collar and taking rest for three to six weeks; 12 twice weekly sessions of physiotherapy and home exercises for six weeks; or continuation of daily activities as much as possible without specific treatment (control group). MAIN OUTCOME MEASURES: Time course of changes in pain scores for arm and neck pain on a 100 mm visual analogue scale and in the neck disability index during the first six weeks. RESULTS: In the wait and see group, arm pain diminished by 3 mm/week on the visual analogue scale (beta=-3.1 mm, 95% confidence interval -4.0 to -2.2 mm) and by 19 mm in total over six weeks. Patients who were treated with cervical collar or physiotherapy achieved additional pain reduction (collar: beta=-1.9 mm, -3.3 to -0.5 mm; physiotherapy: beta=-1.9, -3.3 to -0.8), resulting in an extra pain reduction compared with the control group of 12 mm after six weeks. In the wait and see group, neck pain did not decrease significantly in the first six weeks (beta=-0.9 mm, -2.0 to 0.3). Treatment with the collar resulted in a weekly reduction on the visual analogue scale of 2.8 mm (-4.2 to -1.3), amounting to 17 mm in six weeks, whereas physiotherapy gave a weekly reduction of 2.4 mm (-3.9 to -0.8) resulting in a decrease of 14 mm after six weeks. Compared with a wait and see policy, the neck disability index showed a significant change with the use of the collar and rest (beta=-0.9 mm, -1.6 to -0.1) and a non-significant effect with physiotherapy and home exercises. CONCLUSION: A semi-hard cervical collar and rest for three to six weeks or physiotherapy accompanied by home exercises for six weeks reduced neck and arm pain substantially compared with a wait and see policy in the early phase of cervical radiculopathy. Trial registration Clinical trials NCT00129714. PMID- 19812132 TI - The Israeli Medical Association and doctors' complicity in torture. PMID- 19812133 TI - Antiviral drugs should not be used to treat Bell's palsy, Cochrane analysis concludes. PMID- 19812136 TI - Live and let die. PMID- 19812137 TI - Audit shows large regional variations in uptake of breast reconstruction after mastectomy. PMID- 19812138 TI - Life expectancy differs by 30 years between richest and poorest countries, report shows. PMID- 19812139 TI - UN seeks to boost access to health care for millions of refugees and internally displaced people. PMID- 19812140 TI - Cervarix: definitely not the new MMR. PMID- 19812143 TI - A breathless woman with asthma. PMID- 19812146 TI - Hemagglutinin-dependent tropism of H5N1 avian influenza virus for human endothelial cells. AB - Although current H5N1 highly pathogenic avian influenza viruses (HPAIV) are inefficiently transmitted to humans, infected individuals can suffer from severe disease, often progressing rapidly to acute respiratory distress syndrome and multiorgan failure. This is in contrast with the situation with human influenza viruses, which in immunocompetent individuals usually cause only a respiratory disease which is less aggressive than that observed with avian H5N1 viruses. While the biological basis of inefficient transmission is well documented, the mechanisms by which the H5N1 viruses cause fatal disease remain unclear. In the present study, we demonstrate that human pulmonary microvascular endothelial cells (hPMEC) had a clearly higher susceptibility to infection by H5N1 HPAIV than to infection by human influenza viruses. This was measurable by de novo intracellular nucleoprotein production and virus replication. It was also related to a relatively higher binding capacity to cellular receptors. After infection of hPMEC, cell activation markers E-selectin and P-selectin were upregulated, and the proinflammatory cytokines interleukin-6 and beta interferon were secreted. H5N1 virus infection was also associated with an elevated rate of cell death. Reverse genetics analyses demonstrated a major role for the viral hemagglutinin in this cell tropism. Overall, avian H5N1 viruses have a particular receptor specificity targeting endothelial cells that is different from human influenza viruses, and this H5N1 receptor specificity could contribute to disease pathogenesis. PMID- 19812147 TI - Effects of acute and chronic murine norovirus infections on immune responses and recovery from Friend retrovirus infection. AB - Murine norovirus (MNV) is a highly infectious but generally nonpathogenic agent that is commonly found in research mouse colonies in both North America and Europe. In the present study, the effects of acute and chronic infections with MNV on immune responses and recovery from concurrent Friend virus (FV) infections were investigated. No significant differences in T-cell or NK-cell responses, FV neutralizing antibody responses, or long-term recovery from FV infection were observed. We conclude that concurrent MNV infections had no major impacts on FV infections. PMID- 19812148 TI - Effects of major capsid proteins, capsid assembly, and DNA cleavage/packaging on the pUL17/pUL25 complex of herpes simplex virus 1. AB - The U(L)17 and U(L)25 proteins (pU(L)17 and pU(L)25, respectively) of herpes simplex virus 1 are located at the external surface of capsids and are essential for DNA packaging and DNA retention in the capsid, respectively. The current studies were undertaken to determine whether DNA packaging or capsid assembly affected the pU(L)17/pU(L)25 interaction. We found that pU(L)17 and pU(L)25 coimmunoprecipitated from cells infected with wild-type virus, whereas the major capsid protein VP5 (encoded by the U(L)19 gene) did not coimmunoprecipitate with these proteins under stringent conditions. In addition, pU(L)17 (i) coimmunoprecipitated with pU(L)25 in the absence of other viral proteins, (ii) coimmunoprecipitated with pU(L)25 from lysates of infected cells in the presence or absence of VP5, (iii) did not coimmunoprecipitate efficiently with pU(L)25 in the absence of the triplex protein VP23 (encoded by the U(L)18 gene), (iv) required pU(L)25 for proper solubilization and localization within the viral replication compartment, (v) was essential for the sole nuclear localization of pU(L)25, and (vi) required capsid proteins VP5 and VP23 for nuclear localization and normal levels of immunoreactivity in an indirect immunofluorescence assay. Proper localization of pU(L)25 in infected cell nuclei required pU(L)17, pU(L)32, and the major capsid proteins VP5 and VP23, but not the DNA packaging protein pU(L)15. The data suggest that VP23 or triplexes augment the pU(L)17/pU(L)25 interaction and that VP23 and VP5 induce conformational changes in pU(L)17 and pU(L)25, exposing epitopes that are otherwise partially masked in infected cells. These conformational changes can occur in the absence of DNA packaging. The data indicate that the pU(L)17/pU(L)25 complex requires multiple viral proteins and functions for proper localization and biochemical behavior in the infected cell. PMID- 19812149 TI - A new genetic vaccine platform based on an adeno-associated virus isolated from a rhesus macaque. AB - We created a hybrid adeno-associated virus (AAV) from two related rhesus macaque isolates, called AAVrh32.33, and evaluated it as a vaccine carrier for human immunodeficiency virus type 1 (HIV-1) and type A influenza virus antigens. The goal was to overcome the limitations of vaccines based on other AAVs, which generate dysfunctional T-cell responses and are inhibited by antibodies found in human sera. Injection of a Gag-expressing AAVrh32.33 vector into mice resulted in a high-quality CD8(+) T-cell response. The resulting Gag-specific T cells express multiple cytokines at high levels, including interleukin-2, with many having memory phenotypes; a subsequent boost with an adenovirus vector yielded a brisk expansion of Gag-specific T cells. A priming dose of AAVrh32.33 led to high levels of Gag antibodies, which exceed levels found after injection of adenovirus vectors. Importantly, passive transfer of pooled human immunoglobulin into mice does not interfere with the efficacy of AAVrh32.33 expressing nucleoproteins from influenza virus, as measured by protection to a lethal dose of influenza virus, which is consistent with the very low seroprevalence to this virus in humans. Studies of macaques with vectors expressing gp140 from HIV-1 (i.e., with AAVrh32.33 as the prime and simian adenovirus type 24 as the boost) demonstrated results similar to those for mice with high-level and high-quality CD8(+) T-cell responses to gp140 and high-titered neutralizing antibodies to homologous HIV-1. The biology of this novel AAV hybrid suggests that it should be a preferred genetic vaccine carrier, capable of generating robust T- and B-cell responses. PMID- 19812150 TI - Heparan sulfate proteoglycans are required for cellular binding of the hepatitis E virus ORF2 capsid protein and for viral infection. AB - The hepatitis E virus (HEV), a nonenveloped RNA virus, is the causative agent of hepatitis E. The mode by which HEV attaches to and enters into target cells for productive infection remains unidentified. Open reading frame 2 (ORF2) of HEV encodes its major capsid protein, pORF2, which is likely to have the determinants for virus attachment and entry. Using an approximately 56-kDa recombinant pORF2 that can self-assemble as virus-like particles, we demonstrated that cell surface heparan sulfate proteoglycans (HSPGs), specifically syndecans, play a crucial role in the binding of pORF2 to Huh-7 liver cells. Removal of cell surface heparan sulfate by enzymatic (heparinase) or chemical (sodium chlorate) treatment of cells or competition with heparin, heparan sulfate, and their oversulfated derivatives caused a marked reduction in pORF2 binding to the cells. Syndecan-1 is the most abundant proteoglycan present on these cells and, hence, plays a key role in pORF2 binding. Specificity is likely to be dictated by well-defined sulfation patterns on syndecans. We show that pORF2 binds syndecans predominantly via 6-O sulfation, indicating that binding is not entirely due to random electrostatic interactions. Using an in vitro infection system, we also showed a marked reduction in HEV infection of heparinase-treated cells. Our results indicate that, analogous to some enveloped viruses, a nonenveloped virus like HEV may have also evolved to use HSPGs as cellular attachment receptors. PMID- 19812151 TI - Characterization of a newly identified 35-amino-acid component of the vaccinia virus entry/fusion complex conserved in all chordopoxviruses. AB - The original annotation of the vaccinia virus (VACV) genome was limited to open reading frames (ORFs) of at least 65 amino acids. Here, we characterized a 35 amino-acid ORF (O3L) located between ORFs O2L and I1L. ORFs similar in length to O3L were found at the same genetic locus in all vertebrate poxviruses. Although amino acid identities were low, the presence of a characteristic N-terminal hydrophobic domain strongly suggested that the other poxvirus genes were orthologs. Further studies demonstrated that the O3 protein was expressed at late times after infection and incorporated into the membrane of the mature virion. An O3L deletion mutant was barely viable, producing tiny plaques and a 3-log reduction in infectious progeny. A mutant VACV with a regulated O3L gene had a similar phenotype in the absence of inducer. There was no apparent defect in virus morphogenesis, though O3-deficient virus had low infectivity. The impairment was shown to be at the stage of virus entry, as cores were not detected in the cytoplasm after virus adsorption. Furthermore, O3-deficient virus did not induce fusion of infected cells when triggered by low pH. These characteristics are hallmarks of a group of proteins that form the entry/fusion complex (EFC). Affinity purification experiments demonstrated an association of O3 with EFC proteins. In addition, the assembly or stability of the EFC was impaired when expression of O3 was repressed. Thus, O3 is the newest recognized component of the EFC and the smallest VACV protein shown to have a function. PMID- 19812152 TI - Molecular characterization of a novel gammaretrovirus in killer whales (Orcinus orca). AB - There are currently no published data documenting the presence of retroviruses in cetaceans, though the occurrences of cancers and immunodeficiency states suggest the potential. We examined tissues from adult killer whales and detected a novel gammaretrovirus by degenerate PCR. Reverse transcription-PCR also demonstrated tissue and serum expression of retroviral mRNA. The full-length sequence of the provirus was obtained by PCR, and a TaqMan-based copy number assay did not demonstrate evidence of productive infection. PCR on blood samples from 11 healthy captive killer whales and tissues from 3 free-ranging animals detected the proviral DNA in all tissues examined from all animals. A survey of multiple cetacean species by PCR for gag, pol, and env sequences showed homologs of this virus in the DNA of eight species of delphinids, pygmy and dwarf sperm whales, and harbor porpoises, but not in beluga or fin whales. Analysis of the bottlenose dolphin genome revealed two full-length proviral sequences with 97.4% and 96.9% nucleotide identity to the killer whale gammaretrovirus. The results of single cell PCR on killer whale sperm and Southern blotting are also consistent with the conclusion that the provirus is endogenous. We suggest that this gammaretrovirus entered the delphinoid ancestor's genome before the divergence of modern dolphins or that an exogenous variant existed following divergence that was ultimately endogenized. However, the transcriptional activity demonstrated in tissues and the nearly intact viral genome suggest a more recent integration into the killer whale genome, favoring the latter hypothesis. The proposed name for this retrovirus is killer whale endogenous retrovirus. PMID- 19812153 TI - Identification of hepatitis C virus NS5A inhibitors. AB - Using a cell-based replicon screen, we identified a class of compounds with a thiazolidinone core structure as inhibitors of hepatitis C virus (HCV) replication. The concentration of one such compound, BMS-824, that resulted in a 50% inhibition of HCV replicon replication was approximately 5 nM, with a therapeutic index of >10,000. The compound showed good specificity for HCV, as it was not active against several other RNA and DNA viruses. Replicon cells resistant to BMS-824 were isolated, and mutations were identified. A combination of amino acid substitutions of leucine to valine at residue 31 (L31V) and glutamine to leucine at residue 54 (Q54L) in NS5A conferred resistance to this chemotype, as did a single substitution of tyrosine to histidine at amino acid 93 (Y93H) in NS5A. To further explore the region(s) of NS5A involved in inhibitor sensitivity, genotype-specific NS5A inhibitors were used to evaluate a series of genotype 1a/1b hybrid replicons. Our results showed that, consistent with resistance mapping, the inhibitor sensitivity domain also mapped to the N terminus of NS5A, but it could be distinguished from the key resistance sites. In addition, we demonstrated that NS5A inhibitors, as well as an active-site inhibitor that specifically binds NS3 protease, could block the hyperphosphorylation of NS5A, which is believed to play an essential role in the viral life cycle. Clinical proof of concept has recently been achieved with derivatives of these NS5A inhibitors, indicating that small molecules targeting a nontraditional viral protein like NS5A, without any known enzymatic activity, can also have profound antiviral effects on HCV-infected subjects. PMID- 19812154 TI - Reconstitution of the ancestral glycoprotein of human endogenous retrovirus k and modulation of its functional activity by truncation of the cytoplasmic domain. AB - Endogenous retroviruses present in the human genome provide a rich record of ancient infections. All presently recognized elements, including the youngest and most intact proviruses of the human endogenous retrovirus K(HML-2) [HERV-K(HML 2)] family, have suffered postinsertional mutations during their time of chromosomal residence, and genes encoding the envelope glycoprotein (Env) have not been spared these mutations. In this study, we have, for the first time, reconstituted an authentic Env of a HERV-K(HML-2) provirus by back mutation of putative postinsertional amino acid changes of the protein encoded by HERV-K113. Aided by codon-optimized expression, we demonstrate that the reconstituted Env regained its ability to be incorporated into retroviral particles and to mediate entry. The original ancient HERV-K113 Env was synthesized as a moderately glycosylated gp95 precursor protein cleaved into surface and transmembrane (TM) subunits. Of the nine N-linked oligosaccharides, four are part of the TM subunit, contributing 15 kDa to its apparent molecular mass of 41 kDa. The carbohydrates, as well as the cytoplasmic tail, are critical for efficient intracellular trafficking, processing, stability, and particle incorporation. Whereas deletions of the carboxy-terminal 6 residues completely abrogated cleavage and virion association, more extensive truncations slightly enhanced incorporation but dramatically increased the ability to mediate entry of pseudotyped lentiviruses. Although the first HERV-K(HML-2) elements infected human ancestors about 30 million years ago, our findings indicate that their glycoproteins are in most respects remarkably similar to those of classical contemporary retroviruses and can still mediate efficient entry into mammalian cells. PMID- 19812155 TI - Physical requirements and functional consequences of complex formation between the cytomegalovirus IE1 protein and human STAT2. AB - Our previous work has shown that efficient evasion from type I interferon responses by human cytomegalovirus (hCMV) requires expression of the 72-kDa immediate-early 1 (IE1) protein. It has been suggested that IE1 inhibits interferon signaling through intranuclear sequestration of the signal transducer and activator of transcription 2 (STAT2) protein. Here we show that physical association and subnuclear colocalization of IE1 and STAT2 depend on short acidic and serine/proline-rich low-complexity motifs in the carboxy-terminal region of the 491-amino-acid viral polypeptide. These motifs compose an essential core (amino acids 373 to 420) and an adjacent ancillary site (amino acids 421 to 445) for STAT2 interaction that are predicted to form part of a natively unstructured domain. The presence of presumably "disordered" carboxy-terminal domains enriched in low-complexity motifs is evolutionarily highly conserved across all examined mammalian IE1 orthologs, and the murine cytomegalovirus IE1 protein appears to interact with STAT2 just like the human counterpart. A recombinant hCMV specifically mutated in the IE1 core STAT2 binding site displays hypersensitivity to alpha interferon, delayed early viral protein accumulation, and attenuated growth in fibroblasts. However, replication of this mutant virus is specifically restored by knockdown of STAT2 expression. Interestingly, complex formation with STAT2 proved to be entirely separable from disruption of nuclear domain 10 (ND10), another key activity of IE1. Finally, our results demonstrate that IE1 counteracts the antiviral interferon response and promotes viral replication by at least two distinct mechanisms, one depending on sequestration of STAT2 and the other one likely involving ND10 interaction. PMID- 19812157 TI - Insights into the evolutionary history of an emerging livestock pathogen: porcine circovirus 2. AB - Porcine circovirus 2 (PCV2) is the primary etiological agent of postweaning multisystemic wasting syndrome (PMWS), one of the most economically important emerging swine diseases worldwide. Virulent PCV2 was first identified following nearly simultaneous outbreaks of PMWS in North America and Europe in the 1990s and has since achieved global distribution. However, the processes responsible for the emergence and spread of PCV2 remain poorly understood. Here, phylogenetic and cophylogenetic inferences were utilized to address key questions on the time scale, processes, and geographic diffusion of emerging PCV2. The results of these analyses suggest that the two genotypes of PCV2 (PCV2a and PCV2b) are likely to have emerged from a common ancestor approximately 100 years ago and have been on independent evolutionary trajectories since that time, despite cocirculating in the same host species and geographic regions. The patterns of geographic movement of PCV2 that we recovered appear to mimic those of the global pig trade and suggest that the movement of asymptomatic animals is likely to have facilitated the rapid spread of virulent PCV2 around the globe. We further estimated the rate of nucleotide substitution for PCV2 to be on the order of 1.2 x 10(-3) substitutions/site/year, the highest yet recorded for a single-stranded DNA virus. This high rate of evolution may allow PCV2 to maintain evolutionary dynamics closer to those of single-stranded RNA viruses than to those of double stranded DNA viruses, further facilitating the rapid emergence of PCV2 worldwide. PMID- 19812158 TI - The canine papillomavirus e5 protein signals from the endoplasmic reticulum. AB - The recently discovered Canis familiaris papillomavirus (PV) type 2 (CfPV2) provides a unique opportunity to study PV gene functions in vitro and in vivo. Unlike the previously characterized canine oral PV, CfPV2 contains an E5 open reading frame and is associated with progression to squamous cell carcinoma. In the current study, we have expressed and characterized the CfPV2-encoded E5 protein, a small, hydrophobic, 41-amino-acid polypeptide. We demonstrate that, similar to the E5 protein from high-risk human PV type 16, the CfPV2 E5 protein is localized in the endoplasmic reticulum (ER) and that its expression decreases keratinocyte proliferation and cell life span. E5 expression also increases the percentage of cells in the G(1) phase of the cell cycle, with a concomitant decrease in the percentage of cells in S phase. To identify a potential mechanism for E5-mediated growth inhibition from the ER, we developed a real-time PCR method to quantify the splicing of XBP1 mRNA as a measure of ER stress. We found that the CfPV2 E5 protein induced ER stress and that this, as well as the observed growth inhibition, is tempered significantly by coexpression of the CfPV2 E6 and E7 genes. It is possible that the spatial/temporal regulation of E6/E7 gene expression during keratinocyte differentiation might therefore modulate E5 activity and ER stress. PMID- 19812159 TI - Importance of covalent and noncovalent SUMO interactions with the major human cytomegalovirus transactivator IE2p86 for viral infection. AB - The major transactivator protein IE2p86 of human cytomegalovirus (HCMV) has previously been shown to undergo posttranslational modification by the covalent attachment of SUMO proteins, termed SUMOylation, which occurs at two lysine residues located at amino acid positions 175 and 180. Mutation of the acceptor lysines resulted in the abrogation of IE2p86 SUMOylation in mammalian cells and a strong reduction of IE2p86-mediated transactivation. In this paper, we identify an additional SUMO interaction motif (SIM) within IE2p86, which mediates noncovalent binding to SUMO, as shown by yeast two-hybrid analyses. Transient expression experiments revealed that an IE2p86 SIM mutant exhibited significantly reduced SUMOylation, strongly suggesting that noncovalent SUMO interactions affect the efficacy of covalent SUMO coupling. In order to define the relevance of IE2p86 SUMO interactions for viral replication, recombinant viruses originating from two different HCMV strains (AD169 and VR1814) were generated. Analysis of viruses expressing SUMOylation-negative IE2p86 revealed strongly impaired replication due to reduced viral DNA and protein accumulation, as well as diminished initiation of immediate-early gene expression. The additional introduction of the SIM mutation into the viral genome did not further compromise viral replication but resulted in altered expression of viral proteins at late times postinfection. In summary, this paper clearly shows that IE2p86 SUMOylation is necessary for efficient replication of the HCMV laboratory strain AD169 and the clinical isolate VR1814 and thus for the in vivo function of this viral transcription factor. PMID- 19812160 TI - Recognizing the top 20 peer reviewers for the Journal of Virology. PMID- 19812161 TI - Conversion of VPg into VPgpUpUOH before and during poliovirus negative-strand RNA synthesis. AB - There are two protein primers involved in picornavirus RNA replication, VPg, the viral protein of the genome, and VPgpUpU(OH). A cis-acting replication element (CRE) within the open reading frame of poliovirus (PV) RNA allows the viral RNA dependent RNA polymerase 3D(Pol) to catalyze the conversion of VPg into VPgpUpU(OH). In this study, we used preinitiation RNA replication complexes (PIRCs) to determine when CRE-dependent VPg uridylylation occurs relative to the sequential synthesis of negative- and positive-strand RNA. Guanidine HCl (2 mM), a reversible inhibitor of PV 2C(ATPase), prevented CRE-dependent VPgpUpU(OH) synthesis and the initiation of negative-strand RNA synthesis. VPgpUpU(OH) and nascent negative-strand RNA molecules were synthesized coincident in time following the removal of guanidine, consistent with PV RNA functioning simultaneously as a template for CRE-dependent VPgpUpU(OH) synthesis and negative strand RNA synthesis. The amounts of [(32)P]UMP incorporated into VPgpUpU(OH) and negative-strand RNA products indicated that 100 to 400 VPgpUpU(OH) molecules were made coincident in time with each negative-strand RNA. 3'-dCTP inhibited the elongation of nascent negative-strand RNAs without affecting CRE-dependent VPg uridylylation. A 3' nontranslated region mutation which inhibited negative-strand RNA synthesis did not inhibit CRE-dependent VPg uridylylation. Together, the data implicate 2C(ATPase) in the mechanisms whereby PV RNA functions as a template for reiterative CRE-dependent VPg uridylylation before and during negative-strand RNA synthesis. PMID- 19812162 TI - Determinants of the hepatitis C virus nonstructural protein 2 protease domain required for production of infectious virus. AB - The hepatitis C virus (HCV) nonstructural protein 2 (NS2) is a dimeric multifunctional hydrophobic protein with an essential but poorly understood role in infectious virus production. We investigated the determinants of NS2 function in the HCV life cycle. On the basis of the crystal structure of the postcleavage form of the NS2 protease domain, we mutated conserved features and analyzed the effects of these changes on polyprotein processing, replication, and infectious virus production. We found that mutations around the protease active site inhibit viral RNA replication, likely by preventing NS2-3 cleavage. In contrast, alterations at the dimer interface or in the C-terminal region did not affect replication, NS2 stability, or NS2 protease activity but decreased infectious virus production. A comprehensive deletion and mutagenesis analysis of the C terminal end of NS2 revealed the importance of its C-terminal leucine residue in infectious particle production. The crystal structure of the NS2 protease domain shows that this C-terminal leucine is locked in the active site, and mutation or deletion of this residue could therefore alter the conformation of NS2 and disrupt potential protein-protein interactions important for infectious particle production. These studies begin to dissect the residues of NS2 involved in its multiple essential roles in the HCV life cycle and suggest NS2 as a viable target for HCV-specific inhibitors. PMID- 19812163 TI - Hepatitis C virus (HCV) proteins induce NADPH oxidase 4 expression in a transforming growth factor beta-dependent manner: a new contributor to HCV induced oxidative stress. AB - Viral hepatitis-induced oxidative stress accompanied by increased levels of transforming growth factor beta (TGF-beta) and hepatic fibrosis are hallmarks of hepatitis C virus (HCV) infection. The mechanisms of redox regulation in the pathogenesis of HCV-induced liver disease are not clearly understood. The results of our current studies suggest that reactive oxygen species (ROS) derived from Nox4, a member of the NADPH oxidase (Nox) family, could play a role in HCV induced liver disease. We found that the expression of HCV (genotype 1a) cDNA constructs (full-length and subgenomic), core protein alone, viral RNA, or replicating HCV (JFH-AM2) induced Nox4 mRNA expression and ROS generation in human hepatocyte cell lines (Huh-7, Huh-7.5, HepG2, and CHL). Conversely, hepatocytes expressing Nox4 short hairpin RNA (shRNA) or an inactive dominant negative form of Nox4 showed decreased ROS production when cells were transfected with HCV. The promoters of both human and murine Nox4 were used to demonstrate transcriptional regulation of Nox4 mRNA by HCV, and a luciferase reporter tied to an approximately 2-kb promoter region of Nox4 identified HCV-responsive regulatory regions modulating the expression of Nox4. Furthermore, the human Nox4 promoter was responsive to TGF-beta1, and the HCV core-dependent induction of Nox4 was blocked by antibody against TGF-beta or the expression of dominant negative TGF-beta receptor type II. These findings identified HCV as a regulator of Nox4 gene expression and subsequent ROS production through an autocrine TGF beta-dependent mechanism. Collectively, these data provide evidence that HCV induced Nox4 contributes to ROS production and may be related to HCV-induced liver disease. PMID- 19812164 TI - Early, active, and specific localization of herpes simplex virus type 1 gM to nuclear membranes. AB - Thirteen different glycoproteins are incorporated into mature herpes simplex virus type 1 (HSV-1) virions. Five of them play important roles during entry, while others intervene during egress of the virus. Although HSV-1 gM is not essential in cell culture, its deletion reduces viral yields and promotes syncytium formation. Furthermore, gM is conserved among herpesviruses, is essential for several of them, and can redirect the gD and gH/gL viral glycoproteins from the cell surface to the trans-Golgi network, where gM presumably modulates final capsid envelopment. Late in infection, gM reaches the nuclear envelope and decorates perinuclear virions. This process seemingly requires U(L)31 and U(L)34 and occurs when several markers of the trans-Golgi network have relocalized to the nucleus. However, the precise mechanism of gM nuclear targeting is unclear. We now report that gM is quickly and specifically targeted to nuclear membranes in a virus-dependent manner. This occurs prior to the HSV-1-induced reorganization of the trans-Golgi network and before gM enters the secretory pathway. The presence of a high-mannose glycosylation pattern on gM further corroborated these findings. While gM was targeted to the inner nuclear membrane early in infection, its partners gD, gH, gN, VP22, U(L)31, and U(L)34 did not colocalize with gM. These data suggest that nuclear gM fulfills an early nuclear function that is independent of its known interaction partners and its function in viral egress. PMID- 19812165 TI - Binding of herpes simplex virus glycoprotein B (gB) to paired immunoglobulin-like type 2 receptor alpha depends on specific sialylated O-linked glycans on gB. AB - Paired immunoglobulin-like type 2 receptor alpha (PILRalpha) is an inhibitory receptor expressed on both hematopoietic and nonhematopoietic cells. Its binding to a cellular ligand, CD99, depends on the presence of sialylated O-linked glycans on CD99. Glycoprotein B (gB) of herpes simplex virus type 1 (HSV-1) binds to PILRalpha, and this association is involved in HSV-1 infection. Here, we found that the presence of sialylated O-glycans on gB is required for gB to associate with PILRalpha. Furthermore, we identified two threonine residues on gB that are essential for the addition of the principal O-glycans acquired by gB and that are also essential for the binding of PILRalpha to gB. PMID- 19812166 TI - Downregulation of the T-cell receptor by human immunodeficiency virus type 2 Nef does not protect against disease progression. AB - Chronic immune activation is thought to play a major role in human immunodeficiency virus (HIV) pathogenesis, but the relative contributions of multiple factors to immune activation are not known. One proposed mechanism to protect against immune activation is the ability of Nef proteins from some HIV and simian immunodeficiency virus strains to downregulate the T-cell receptor (TCR)-CD3 complex of the infected cell, thereby reducing the potential for deleterious activation. HIV type 1 (HIV-1) Nef has lost this property. In contrast to HIV-1, HIV-2 infection is characterized by a marked disparity in the disease course, with most individuals maintaining a normal life span. In this study, we examined the relationship between the ability of HIV-2 Nef proteins to downregulate the TCR and immune activation, comparing progressors and nonprogressors. Representative Nef variants were isolated from 28 HIV-2-infected individuals. We assessed their abilities to downregulate the TCR from the surfaces of CD4 T cells. In the same individuals, the activation of peripheral lymphocytes was evaluated by measurement of the expression levels of HLA-DR and CD38. We observed a striking correlation of the TCR downregulation efficiency of HIV-2 Nef variants with immune activation in individuals with a low viral load. This strongly suggests that Nef expression can influence the activation state of the immune systems of infected individuals. However, the efficiency of TCR downregulation by Nef was not reduced in progressing individuals, showing that TCR downregulation does not protect against progression in HIV-2 infection. PMID- 19812167 TI - High frequencies of virus-specific CD8+ T-cell precursors. AB - A productive CD8(+) T-cell response to a viral infection requires rapid division and proliferation of virus-specific CD8(+) T cells. Tetramer-based enrichment assays have recently given estimates of the numbers of peptide-major histocompatibility complex-specific CD8(+) T cells in naive mice, but precursor frequencies for entire viruses have been examined only by using in vitro limiting dilution assays (LDAs). To examine CD8(+) T-cell precursor frequencies for whole viruses, we developed an in vivo LDA and found frequencies of naive CD8(+) T-cell precursors of 1 in 1,444 for vaccinia virus (VV) ( approximately 13,850 VV specific CD8(+) T cells per mouse) and 1 in 2,958 for lymphocytic choriomeningitis virus (LCMV) ( approximately 6,761 LCMV-specific CD8(+) T cells per mouse) in C57BL/6J mice. In mice immune to VV, the number of VV-specific precursors, not surprisingly, dramatically increased to 1 in 13 ( approximately 1,538,462 VV-specific CD8(+) T cells per mouse), consistent with estimates of VV specific memory T cells. In contrast, precursor numbers for LCMV did not increase in VV-immune mice (1 in 4,562, with approximately 4,384 LCMV-specific CD8(+) T cells per VV-immune mouse). Using H-2D(b)-restricted LCMV GP33-specific P14 transgenic T cells, we found that, after donor T-cell take was accounted for, approximately every T cell transferred underwent a full proliferative expansion in response to LCMV infection. This high efficiency was also seen with memory populations, suggesting that most antigen-specific T cells will proliferate extensively at a limiting dilution in response to infections. These results show that frequencies of naive and memory CD8(+) T cell precursors for whole viruses can be remarkably high. PMID- 19812168 TI - Characterization of viral and human RNAs smaller than canonical MicroRNAs. AB - Recently identified small (20 to 40 bases) RNAs, such as microRNAs (miRNAs) and endogenous small interfering RNAs (siRNAs) participate in important cellular pathways. In this report, we systematically characterized several novel features of human and viral RNA products smaller than miRNAs. We found that Kaposi sarcoma associated herpesvirus K12-1 miRNA (23 bases) associates with a distinct, unusually small (17-base) RNA (usRNA) that can effectively downregulate a K12-1 miRNA target, human RAD21, suggesting that stable degradation-like products may also contribute to gene regulation. High-throughput sequencing reveals a diverse set of human miRNA-derived usRNAs and other non-miRNA-derived usRNAs. Human miRNA derived usRNAs preferentially match to 5' ends of miRNAs and are also more likely to associate with the siRNA effector protein Ago2 than with Ago1. Many non-miRNA derived usRNAs associate with Ago proteins and also frequently contain C-rich 3' specific motifs that are overrepresented in comparison to Piwi-interacting RNAs and transcription start site-associated RNAs. We postulate that approximately 30% of usRNAs could have evolved to participate in biological processes, including gene silencing. PMID- 19812169 TI - Rift valley fever virus L protein forms a biologically active oligomer. AB - Rift Valley fever virus (RVFV) (genus Phlebovirus, family Bunyaviridae) causes mosquito-borne epidemic diseases in humans and livestock. The virus carries three RNA segments, L, M, and S, of negative or ambisense polarity. L protein, an RNA dependent RNA polymerase, encoded in the L segment, and N protein, encoded in the S segment, exert viral RNA replication and transcription. Coexpression of N, hemagglutinin (HA)-tagged L, and viral minigenome resulted in minigenome replication and transcription, a finding that demonstrated HA-tagged L was biologically active. Likewise L tagged with green fluorescent protein (GFP) was biologically competent. Coimmunoprecipitation analysis using extracts from cells coexpressing HA-tagged L and GFP-tagged L showed the formation of an L oligomer. Bimolecular fluorescence complementation analysis and coimmunoprecipitation studies demonstrated the formation of an intermolecular L-L interaction through its N-terminal and C-terminal regions and also suggested an intramolecular association between the N-terminal and C-terminal regions of L protein. A biologically inactive L mutant, in which the conserved signature SDD motif was replaced by the amino acid residues GNN, exhibited a dominant negative phenotype when coexpressed with wild-type L in the minigenome assay system. Expression of this mutant L also inhibited viral gene expression in virus-infected cells. These data provided compelling evidence for the importance of oligomerization of RVFV L protein for its polymerase activity. PMID- 19812170 TI - Detection of novel sequences related to african Swine Fever virus in human serum and sewage. AB - The family Asfarviridae contains only a single virus species, African swine fever virus (ASFV). ASFV is a viral agent with significant economic impact due to its devastating effects on populations of domesticated pigs during outbreaks but has not been reported to infect humans. We report here the discovery of novel viral sequences in human serum and sewage which are clearly related to the asfarvirus family but highly divergent from ASFV. Detection of these sequences suggests that greater genetic diversity may exist among asfarviruses than previously thought and raises the possibility that human infection by asfarviruses may occur. PMID- 19812171 TI - Cumulative effects and predictive value of common obesity-susceptibility variants identified by genome-wide association studies. AB - BACKGROUND: Large-scale genome-wide association studies have identified 12 genetic loci that are robustly associated with body mass index (BMI). OBJECTIVES: We examined associations and compared effect sizes of these newly identified obesity susceptibility loci with various anthropometric traits and assessed their cumulative effects and predictive value for obesity risk. DESIGN: We genotyped 12 single nucleotide polymorphisms (SNPs) from each locus in 20,431 individuals (age: 39-79 y) from the population-based European Prospective Investigation into Cancer and Nutrition (EPIC)-Norfolk cohort. General linear model and logistic regression were used to examine associations, and the area under the receiver operating characteristic curve (AUC) was used to assess the predictive value of these variants for obesity risk. RESULTS: Effect sizes of the risk alleles ranged between 0.058 and 0.329 for BMI (in kg/m(2)), between 0.094 and 0.866 kg for weight, and between 0.085 and 0.819 cm for waist circumference, with rs1121980 (FTO locus) showing the largest effect. Risk alleles of rs7132908 (FAIM2 locus) and rs17782313 (MC4R locus) were also associated with taller height. On average, each additional risk allele was associated with increases of 0.149 in BMI (P = 1.54 x 10(-22)), 0.444 kg in body weight (P = 9.88 x 10(-22)), and 0.357 cm in waist circumference (P = 1.10 x 10(-18)) and 10.8% (P = 9.83 x 10(-16)) and 5.5% (P = 3.38 x 10(-10)) increased risks of obesity and overweight, respectively. All SNPs combined explained 0.9% of BMI variation, with an AUC of 0.574 (95% CI: 0.559, 0.590) for prediction of obesity. CONCLUSIONS: Common variants for BMI have small effects on obesity measures and show different association patterns with anthropometric traits, with the largest effect shown for the FTO locus. These variants have cumulative effects, yet their predictive value for obesity risk is limited. PMID- 19812172 TI - Sugar and fiber intake and type of adiposity: are they related? PMID- 19812173 TI - Prenatal food supplementation fortified with multiple micronutrients increases birth length: a randomized controlled trial in rural Burkina Faso. AB - BACKGROUND: Prenatal multiple micronutrient (MMN) or balanced energy and protein supplementation has a limited effect on birth size of the offspring. OBJECTIVE: The objective was to determine whether a prenatal MMN-fortified food supplement (FFS) improves anthropometric measures at birth compared with supplementation with an MMN pill alone. DESIGN: We conducted a nonblinded, individually randomized controlled trial in 1296 pregnant women in 2 villages in rural Burkina Faso. Supplements were provided on a daily basis, and compliance was closely verified by using a community-based network of home visitors. RESULTS: Anthropometric measures at birth were available for analysis for 87% of the 1175 live singleton deliveries enrolled. After adjustment for gestational age at birth, the FFS group had a significantly higher birth length (+4.6 mm; P = 0.001). FFS supplementation resulted in a modestly higher birth weight (+31 g; P = 0.197). Subgroup analyses showed clinically important treatment effects on birth length (+12.0 mm; P = 0.005) and on birth weight (+111 g; P = 0.133) for underweight [body mass index (in kg/m(2)) <18.5] pregnant women. Women with early pregnancy anemia who received FFS gave birth to longer newborns (+7.3 mm; P = 0.002) than did those who received MMN supplementation. CONCLUSIONS: The provision of FFS to pregnant women resulted in higher birth length than did MMN supplementation. For women with a suboptimal prepregnancy nutritional status, MMN supplementation should be complemented with a balanced energy and protein supplement to produce a clinical effect on birth size. The trial was registered at clinicaltrials.gov as NCT00909974. PMID- 19812174 TI - Metabolic rate analysis of healthy preterm and full-term infants during the first weeks of life. AB - BACKGROUND: Longitudinal data on resting energy expenditure (REE) in extremely immature infants and full-term neonates are scarce but are necessary to understand the energy requirements in neonatal nutrition during the first weeks of life. OBJECTIVE: The aim of the present study was to measure REE and its main components longitudinally during the first weeks of life to quantify their significant determinants. DESIGN: REE was investigated longitudinally over a period of 6 wk in healthy, stable, and growing preterm infants and over 5 wk in full-term neonates by means of indirect calorimetry. RESULTS: A total of 197 infants, including 183 premature infants and 14 full-term neonates, were recruited for the study. REE values increased in all gestational age groups from the first week to 5-6 wk of postnatal age, with the most pronounced increase in the smallest infants (+140%) and the smallest increase in the full-term neonates (+47%). Univariate calculations showed that for each postnatal week, REE increased by 6.93-9.64 kcal x kg(-1) x d(-1) with each additional kcal administered, for an average increase of 0.701 kcal, and increased by 1.78 kcal for each 1 g gain in weight. Postnatal age was the strongest predictor to influence REE (r(2) = 0.727, P < 0.0001). CONCLUSIONS: This study provides comprehensive data on longitudinally determined REE values of healthy premature and full-term infants. Results may serve as a basis for comparative studies that address various disease states as well as different nutritional protocols. PMID- 19812175 TI - Effect of short-term high-protein compared with normal-protein diets on renal hemodynamics and associated variables in healthy young men. AB - BACKGROUND: High-protein diets are effective for weight reduction; however, little is known about the potential adverse renal effects of such diets. OBJECTIVE: The aim of our study was to compare the effect of a high-protein (HP) with a normal-protein (NP) diet on renal hemodynamics and selected clinical chemical factors. DESIGN: We prospectively studied the effect of an HP diet (2.4 g x kg(-1) x d(-1)) with that of an NP diet (1.2 g x kg(-1) x d(-1)) on the glomerular filtration rate (assessed on the basis of sinistrin-an inulin analog clearance) and renal plasma flow (para-aminohippuric acid clearance) by using the constant infusion technique. Filtration fraction and renal vascular resistance were calculated. Twenty-four healthy young men followed the 2 diet protocols for 7 d each in a crossover design. They were individually advised by a dietitian to achieve the planned protein intake by selecting normal foods under isocaloric conditions. Serum and urinary variables and renal hemodynamics were measured on day 7 of both diets. RESULTS: The glomerular filtration rate (NP: 125 +/- 5 mL/min; HP: 141 +/- 8 mL/min; P < 0.001) and filtration fraction (NP: 23 +/- 5%; HP: 28 +/- 5%; P < 0.05) increased significantly with the HP diet. Renal plasma flow was not significantly different between the HP (496 +/- 25 mL/min) and NP (507 +/- 18 mL/min) phases. Renal vascular resistance was not significantly different between the NP (94 +/- 6 mm Hg x mL(-1) x min(-1)) and HP (99 +/- 8 mm Hg x mL(-1) x min(-1)) phases. Blood urea nitrogen, serum uric acid, glucagon, natriuresis, urinary albumin, and urea excretion increased significantly with the HP diet. CONCLUSIONS: A short-term HP diet alters renal hemodynamics and renal excretion of uric acid, sodium, and albumin. More attention should be paid to the potential adverse renal effects of HP diets. PMID- 19812176 TI - {omega}-3 Long-chain polyunsaturated fatty acid intake and 12-y incidence of neovascular age-related macular degeneration and central geographic atrophy: AREDS report 30, a prospective cohort study from the Age-Related Eye Disease Study. AB - BACKGROUND: omega-3 (n-3) Long-chain polyunsaturated fatty acids (LCPUFAs) affect processes implicated in vascular and neural retinal pathogenesis and thus may influence the risk of developing age-related macular degeneration (AMD). OBJECTIVE: We investigated whether omega-3 LCPUFA intake was associated with a reduced likelihood of developing central geographic atrophy (CGA) and neovascular (NV) AMD. DESIGN: We undertook a nested cohort study within a multicenter phase 3 clinical trial, the Age-Related Eye Disease Study (AREDS), to study progression to advanced AMD in 1837 persons at moderate-to-high risk of this condition. The AREDS was designed to assess the clinical course, prognosis, risk factors, and nutrient-based treatments of AMD and ran from November 1992 to December 2005. We obtained baseline data on omega-3 LCPUFA intake with a validated food-frequency questionnaire. Trained fundus graders ascertained AMD status from annual stereoscopic color photographs by using standardized methods at a single reading center across a 12-y period. We applied multivariable repeated-measures logistic regression with the incorporation of generalized estimating equation methods, because this permitted determination of progression to outcome at each visit. RESULTS: Participants who reported the highest omega-3 LCPUFA intake (median: 0.11% of total energy intake) were 30% less likely than their peers to develop CGA and NV AMD. The respective odds ratios were 0.65 (95% CI: 0.45, 0.92; P or=72 years, semantic fluency less than 20 words in 90 s and inability to copy an intersecting pentagons figure, are significant predictors of dementia risk, thus validating our previous findings. In combination, these factors had an odds ratio of 88 for dementia within the first 5 years from diagnosis and may reflect the syndrome of mild cognitive impairment of Parkinson's disease. Phonemic fluency and other frontally based tasks were not associated with dementia risk. MAPT H1/H1 genotype was an independent predictor of dementia risk (odds ratio = 12.1) and the H1 versus H2 haplotype was associated with a 20% increase in transcription of 4-repeat tau in Lewy body disease brains. In contrast, COMT genotype had no effect on dementia, but a significant impact on Tower of London performance, a frontostriatally based executive task, which was dynamic, such that the ability to solve this task changed with disease progression. Hence, we have identified three highly informative predictors of dementia in Parkinson's disease, which can be easily translated into the clinic, and established that MAPT H1/H1 genotype is an important risk factor with functional effects on tau transcription. Our work suggests that the dementing process in Parkinson's disease is predictable and related to tau while frontal-executive dysfunction evolves independently with a more dopaminergic basis and better prognosis. PMID- 19812214 TI - Interaction of the HIV-1 frameshift signal with the ribosome. AB - Ribosomal frameshifting on viral RNAs relies on the mechanical properties of structural elements, often pseudoknots and more rarely stem-loops, that are unfolded by the ribosome during translation. In human immunodeficiency virus (HIV)-1 type B a long hairpin containing a three-nucleotide bulge is responsible for efficient frameshifting. This three-nucleotide bulge separates the hairpin in two domains: an unstable lower stem followed by a GC-rich upper stem. Toeprinting and chemical probing assays suggest that a hairpin-like structure is retained when ribosomes, initially bound at the slippery sequence, were allowed multiple EF-G catalyzed translocation cycles. However, while the upper stem remains intact the lower stem readily melts. After the first, and single step of translocation of deacylated tRNA to the 30 S P site, movement of the mRNA stem-loop in the 5' direction is halted, which is consistent with the notion that the downstream secondary structure resists unfolding. Mechanical stretching of the hairpin using optical tweezers only allows clear identification of unfolding of the upper stem at a force of 12.8 +/- 1.0 pN. This suggests that the lower stem is unstable and may indeed readily unfold in the presence of a translocating ribosome. PMID- 19812215 TI - One-carbon metabolism-genome interactions in folate-associated pathologies. AB - Impairments in folate-mediated 1-carbon metabolism are associated with several common diseases and developmental anomalies including intestinal cancers, vascular disease, cognitive decline, and neural tube defects. The etiology of folate-associated pathologies involves interactions among multiple genetic risk alleles and environmental factors, although the causal mechanisms that define the role of folate and other B-vitamins in these complex disorders remain to be established. Folate and other B-vitamins fundamentally differ from other nutrients that interact with the genome in determining health and disease outcomes in that their interaction is reciprocal. Common gene variants influence the activity of folate-dependent enzymes and anabolic pathways; folate-mediated 1 carbon metabolism is essential for the high-fidelity synthesis of DNA and activated methyl groups that are required for DNA methylation and regulation of chromatin structure. This review focuses on the regulation of folate-mediated 1 carbon metabolism and its role in maintaining genome integrity and on strategies for establishing the metabolic pathways and mechanisms that underlie folate associated pathologies. PMID- 19812217 TI - Overview to symposium "Nutrients and epigenetic regulation of gene expression". AB - The American Society for Nutrition hosted a symposium entitled Nutrients and Epigenetic Regulation of Gene Expression at the Experimental Biology meeting on April 20, 2009, in New Orleans, LA. The symposium was cochaired by Emily Ho from Oregon State University and the Linus Pauling Institute, and Janos Zempleni from the University of Nebraska at Lincoln. The goal of this symposium was to highlight the interactions among nutrients, epigenetics, and disease susceptibility. The symposium featured 4 speakers, each presenting novel insights into mechanisms by which nutrients participate in gene regulation. Janos Zempleni elucidated mechanisms by which the covalent binding of biotin to histones represses transposable elements, thereby enhancing genome stability. Emily Ho shared valuable insights into bioactive food compounds that inhibit histone deacetylases. James Kirkland from the University of Guelph in Canada talked about a niacin-dependent poly(ADP-ribosylation) of histones, an epigenetic mark that is not currently being given full consideration in nutrition. Patrick Stover from Cornell University described the interrelationships among 1-carbon metabolism, DNA methylation, gene silencing, and their influence in the etiology of folate related pathologies. All 4 presentations were videotaped and can be viewed online (www.nutrition.org). PMID- 19812216 TI - Repression of transposable elements by histone biotinylation. AB - Transposable elements constitute >40% of the human genome; transposition of these elements increases genome instability and cancer risk. Epigenetic mechanisms are important for transcriptional repression of retrotransposons, thereby preventing transposition events. Binding of biotin to histones, mediated by holocarboxylase synthetase (HCS), is a novel histone mark that plays a role in gene regulation. Here, we review recent findings that biotinylation of lysine-12 in histone H4 (H4K12bio) is an epigenetic mechanism to repress long terminal repeat (LTR) retrotransposons in human and mouse cell lines, primary cells from human adults, and in Drosophila melanogaster. Further, evidence is summarized that supports a causal relationship between the repression of LTR in H4K12bio-depleted cells and increased production of viral particles, increased frequency of retrotransposition events, and increased frequency of chromosomal abnormalities in mammals and Drosophila. Although HCS interacts physically with histones H3 and H4, the mechanism responsible for targeting HCS to retrotransposons to mediate histone biotinylation is uncertain. We hypothesize that HCS binds specifically to genomic regions rich in methylated cytosines and catalyzes increased biotinylation of histone H4 at lysine-12. Further, we hypothesize that this biotinylation promotes the subsequent dimethylation of lysine-9 in histone H3, resulting in an overall synergistic effect of 3 diet-dependent covalent modifications of histones in the repression of LTR. PMID- 19812218 TI - A metabolite profiling approach to identify biomarkers of flavonoid intake in humans. AB - Flavonoids are phytochemicals that are widespread in the human diet. Despite limitations in their bioavailability, experimental and epidemiological data suggest health benefits of flavonoid consumption. Valid biomarkers of flavonoid intake may be useful for estimating exposure in a range of settings. However, to date, few useful flavonoid biomarkers have been identified. In this study, we used a metabolite profiling approach to examine the aromatic and phenolic profile of plasma and urine of healthy men after oral consumption of 200 mg of the pure flavonoids, quercetin, (-)-epicatechin, and epigallocatechin gallate, which represent major flavonoid constituents in the diet. Following enzymatic hydrolysis, 71 aromatic compounds were quantified in plasma and urine at 2 and 5 h, respectively, after flavonoid ingestion. Plasma concentrations of different aromatic compounds ranged widely, from 0.01 to 10 micromol/L, with variation among volunteers. None of the aromatic compounds was significantly elevated in plasma 2 h after consumption of either flavonoid compared with water placebo. This indicates that flavonoid-derived aromatic compounds are not responsible for the acute physiological effects reported within 2 h in previous human intervention studies involving flavonoids or flavonoid-rich food consumption. These effects are more likely due to absorption of the intact flavonoid. Our urine analysis suggested that urinary 4-ethylphenol, benzoic acid, and 4 ethylbenzoic acid may be potential biomarkers of quercetin intake and 1,3,5 trimethoxybenzene, 4-O-methylgallic acid, 3-O-methylgallic acid, and gallic acid may be potential markers of epigallocatechin gallate intake. Potential biomarkers of (-)-epicatechin were not identified. These urinary biomarkers may provide an accurate indication of flavonoid exposure. PMID- 19812219 TI - Diallyl trisulfide protects rats from carbon tetrachloride-induced liver injury. AB - Alk(en)yl sulfides have been found to be responsible for the anticancer, antithrombotic, and antioxidant effects of garlic. We sought to identify the most potent structure of sulfides that exhibits a hepatoprotective effect against carbon tetrachloride (CCl(4))-induced acute liver injury in rats. Rats were pretreated with diallyl trisulfide (DATS) i.g. at a dose of 500 micromol/kg body weight for 5 d. On d 6, CCl(4) was administered i.g. at a dose of 2.5 mL/kg body weight. Twenty-four hours after CCl(4) administration, rats were killed and plasma and liver samples collected. DATS pretreatment significantly suppressed the CCl(4)-induced elevation of plasma aspartate aminotransferase and alanine aminotransferase activities (P < 0.05). Histological observations supported the hepatoprotective effects. Western blot and spectrophotometric analyses indicated that DATS suppressed cytochrome P450 2E1 activity and its protein level and elevated those of glutathione S-transferase. Dipropyl trisulfide (DPTS), which is a saturated alkyl chain analogue of DATS, did not affect CCl(4)-induced liver toxicity or drug-metabolizing enzymes. These results suggest that hepatoprotective activity of trisulfides is due to their regulation of drug metabolizing enzymes. Furthermore, the effects of 6 kinds of alk(en)yl trisulfides, including DATS and DPTS, on phase II enzyme activity were examined in rats. Alk(en)yl trisulfides were administered i.g. (500 micromol/kg body weight) to rats for 5 d. Only the allyl group-containing DATS and allyl methyl trisulfide enhanced these activities. PMID- 19812220 TI - Body mass index is an important determinant of methylation biomarkers in women of reproductive ages. AB - B vitamin deficiencies lead to moderate hyperhomocysteinemia, which has been associated with health and disease. However, concomitant derangements in cellular methylation, reflected by altered plasma S-adenosylmethionine (SAM) or S adenosylhomocysteine (SAH) concentrations, may be the primary cause. Therefore, we identified determinants of homocysteine, SAM, and SAH concentrations in 336 women, aged 20-48 y, as part of a large study focusing on risk factors for reproductive disorders. Blood was obtained to determine plasma SAM, SAH, and total homocysteine (tHcy), serum vitamin B-12 and folate, RBC folate concentrations, and the related single nucleotide polymorphisms 5,10 methylenetetrahydrofolate reductase (MTHFR) 677C > T and 1298A > C, methionine synthase reductase (MTRR) 66A > G, and nicotinamide N-methyltransferase IVS1-151G > A. Questionnaires provided information on demographics, lifestyles, and nutrient intakes. Correlation coefficients were calculated and multivariable associations were assessed with a general linear model. Serum folate was positively correlated with SAM concentrations (r = 0.159; P = 0.004). Folate and vitamin B-12 were not correlated with SAH concentrations or the SAM:SAH ratio but were inversely correlated with tHcy concentrations (serum folate r = -0.324; RBC folate r = -0.294; vitamin B-12 r = -0.307; P < 0.01). From the multivariable analysis, BMI was the strongest determinant of SAM (standardized beta = 19.145; P < 0.001) and SAH concentrations (standardized beta = 3.241; P = 0.010). MTHFR 677TT (standardized beta = 0.195; P = 0.001), B vitamin supplement use (standardized beta = -0.156; P < 0.001) and dietary protein intake (standardized beta = -0.011; P < 0.001) were the strongest determinants of tHcy concentrations. Thus, the determinants of SAM and SAH differ from those of tHcy concentrations. Given that BMI was a strong determinant of SAM concentrations, it should be included in future studies on cellular methylation. PMID- 19812221 TI - Niacin status impacts chromatin structure. AB - Niacin is required to form NAD and NADP, which are involved in many essential redox reactions in cellular metabolism. In addition, NAD(+) acts as a substrate for a variety of ADP-ribosylation reactions, including poly- and mono-ADP ribosylation of proteins, formation of cyclic ADP-ribose, and the generation of O acetyl-ADP-ribose in deacetylation reactions. These nonredox reactions are critical in the regulation of cellular metabolism, and they are sensitive to dietary niacin status. There are 4 known mechanisms by which ADP-ribosylation reactions directly regulate chromatin structure. These include the covalent modification of histones with poly(ADP-ribose), the extraction of histones from chromatin by noncovalent binding to poly(ADP-ribose) on poly(ADP-ribose) polymerase-1, poly ADP-ribosylation of telomeric repeat-binding factor-1 within telomeres, and deacetylation of histones by the sirtuins. These reactions produce a variety of localized effects in chromatin structure, and altered function in response to changes in niacin status may have dramatic effects on genomic stability, cell division and differentiation, and apoptosis. PMID- 19812222 TI - Dietary sulforaphane, a histone deacetylase inhibitor for cancer prevention. AB - The reversible acetylation of histones is an important mechanism of gene regulation. During prostate cancer progression, specific modifications in acetylation patterns on histones are apparent. Targeting the epigenome, including the use of histone deacetylase (HDAC) inhibitors, is a novel strategy for cancer chemoprevention. Recently, drugs classified as HDAC inhibitors have shown promise in cancer clinical trials. We have previously found that sulforaphane (SFN), a compound found in cruciferous vegetables, inhibits HDAC activity in human colorectal and prostate cancer cells. Based on the similarity of SFN metabolites and other phytochemicals to known HDAC inhibitors, we previously demonstrated that sulforaphane acted as an HDAC inhibitor in the prostate, causing enhanced histone acetylation, derepression of P21 and Bax, and induction of cell cycle arrest/apoptosis, leading to cancer prevention. The ability of SFN to target aberrant acetylation patterns, in addition to effects on phase 2 enzymes, may make it an effective chemoprevention agent. These studies are important because of the potential to qualify or change recommendations for high-risk prostate cancer patients and thereby increase their survival through simple dietary choices incorporating easily accessible foods into their diets. These studies also will provide a strong scientific foundation for future large-scale human clinical intervention studies. PMID- 19812224 TI - Prevention of mother to child transmission of HIV in Burkina Faso: breastfeeding and wet nursing. AB - OBJECTIVE: A survey of the knowledge of women about HIV and breastfeeding. METHOD AND PATIENTS: The study employed a voluntary questionnaire. Data were analyzed using Epi info 6 with chi(2) test and P < 0.05 was considered as statistically significant. RESULTS: Three hundred women agreed to participate in the study. Only 82 realized that HIV could be transmitted by breastmilk. After information about HIV transmission, 37 stated that they would still breastfeed. On the other hand, 220 (74.1%) stated that they would accept breastfeeding by a wet nurse. Two hundred and eighty (69.8%) would accept to serve as the wet nurse for an infant born to an HIV-infected woman. There was an association between acceptance of HIV screening and willingness to breastfeed (P = 0.00206529). CONCLUSION: Appropriate Prevention of Mother-To-Child Transmission (PMTCT) measures must be made available to HIV-infected women. Detailed studies must be performed to evaluate the feasibility of this approach in resource limited settings. PMID- 19812223 TI - Reduced milk triglycerides in mice lacking phosphoenolpyruvate carboxykinase in mammary gland adipocytes and white adipose tissue contribute to the development of insulin resistance in pups. AB - Obesity and type 2 diabetes are growing problems worldwide in adults and children. In this study, we focused on understanding the patterning of insulin resistance as a result of altered perinatal nutrition. We analyzed mice in which the binding site for PPARgamma was deleted from the promoter of the cytosolic phosphoenolpyruvate carboxykinase gene (Pck1) (PPARE(-/-)). We analyzed pups from dams with the same genotype as well as fostered and cross-fostered pups. Pck1 expression and triglyceride concentration in the milk were measured. The PPARE mutation reduced Pck1 expression in white adipose tissue (WAT) to 2.2% of wild type (WT) and reduced Pck1 expression in whole mammary gland tissue to 1% of WT. The female PPARE(-/-) mice had reduced lipid storage in mammary gland adipocytes and in WAT, resulting in a 40% reduction of milk triglycerides during lactation. Pups from PPARE(-/-) dams had insulin resistance as early as 14 d after birth, a condition that persisted into adulthood. WT pups fostered by PPARE(-/-) dams had lower body weights and plasma insulin concentrations compared with WT pups reared by WT dams. PPARE(-/-) pups fostered by WT dams had improved glucose clearance compared with pups raised by PPARE(-/-) dams. PPARE(+/-) and PPARE(-/-) dams also patterned newborn pups for reduced growth and insulin resistance in utero. Thus, the in utero environment and altered nutrition during the perinatal period cause epigenetic changes that persist into adulthood and contribute to the development of insulin resistance. PMID- 19812226 TI - When a predator avoids infected prey: a model-based theoretical study. AB - In this paper we study a predator-prey model with logistic growth in the prey population, where a disease spreads among the prey according to an susceptible infected-susceptible (SIS) epidemic model. The predators do not consume infected prey. After a review of the literature we formulate the basic mathematical model. For simplicity, we work initially with a model involving the fractions of prey susceptible and infected and then translate the results back to the model with absolute numbers. Both local and global stability results are examined. For the model working with absolute numbers, we find six possible equilibria and three important threshold values determining the behaviour of the system. There is always a unique locally stable equilibrium. We make conjectures concerning the global behaviour of the system. Next the effect of predator removal on the ecoepidemiological system is examined. The penultimate section describes numerical simulations using realistic parameter values for a real-life situation. This is humans predating on fish (Atlantic cod) infected by bacterial fin rot. The simulations confirm our conjectures. A discussion concludes the paper. PMID- 19812227 TI - Geriatric oncology. PMID- 19812228 TI - Response of elderly patients with rheumatoid arthritis to methotrexate or TNF inhibitors compared with younger patients. AB - OBJECTIVE: To compare the efficacy of MTX and MTX+TNF inhibitors (TNFis) in elderly patients with RA with that in patients of younger age. METHODS: Data from two large, randomized, controlled, double-blind trials in patients with early RA using adalimumab or infliximab+MTX or MTX alone were obtained and pooled. Composite disease activity indices were calculated at baseline and 1 year of treatment, and compared in groups of patients classified by quartiles of age with the highest age group comprising 61-82 years using analysis of variance or Kruskal-Wallis test. RESULTS: Across all age quartiles, improvement on MTX was similar with respect to changes of composite disease activity indices, assessment of physical function and X-ray progression. Likewise, TNFi+MTX had similar effects across all age groups, but the effects of the combination were more profound than those of MTX monotherapy. Also in 10% of the patients with the highest age, primarily septuagenarians, improvement was seen to a similar degree as in the younger ones. CONCLUSIONS: Responsiveness of elderly patients with RA to MTX or TNFi+MTX is similar to that observed in patients of younger age. PMID- 19812229 TI - Access to training in musculoskeletal ultrasound: a survey of UK rheumatology trainees. PMID- 19812230 TI - Analyses of injury count data: some do's and don'ts. AB - The analysis of injury data requires different considerations from the analysis of other types of outcomes because an individual can experience the outcome many times. When describing injury patterns using numerator-only data (e.g., proportion of upper-extremity injuries vs. lower-extremity injuries), simple comparisons of proportions are inappropriate because 1) individuals are compared with themselves and 2) multiple testing increases the potential for incorrect inference. Bootstrapping (resampling) techniques can be used to determine confidence intervals and whether the frequencies significantly differ across categories. When describing injury rates, the authors suggest plotting the observed injury rate against the number of exposures to obtain a visual representation of the heterogeneity of risk across individuals. Because the distribution of injury rates is often skewed, some research questions may be best addressed by comparing the weighted median injury rates instead of the weighted mean injury rates (which are given by standard formulae). Again, resampling techniques can be used to obtain a null distribution for injury rates in order to determine whether there are subjects who have unexpectedly high injury rates. More advanced analyses are required to account for multiplicity. PMID- 19812231 TI - Comparison of direct and indirect methods of measuring airborne chrysotile fibre concentration. AB - Transmission electron microscopy observations most frequently form a basis for estimating asbestos fibre concentration in the environment and in buildings with asbestos-containing materials. Sampled fibres can be transferred to microscope grids by applying either a direct [ISO (1995) Draft International ISO/DIS 10312. Ambient air. Determination of asbestos fibres. Direct transfer transmission electron microscopy procedure. Geneva, Switzerland: International Standardization Organization] or an indirect [AFNOR (1996) Determination de la concentration en fibres d'amiante par microscopie electronique a transmission-Methode indirecte. Cedex, France: AFNOR, p. 42; ISO (1997) Draft International ISO/DIS 13794. Ambient air. Determination of asbestos fibres. Indirect-transfer transmission electron microscopy procedure. Geneva, Switzerland: International Standardization Organization] method. In the latter case, ISO Standard 13794 recommends filtering calcination residues either on a polycarbonate (PC) filter (PC indirect method) or on a cellulose ester (CE) membrane (CE indirect method). The PC indirect method requires that fibres deposited on a PC filter be covered by a carbon layer, whereas in the CE indirect method, the CE membrane has to be directly processed using a method described in ISO Standard 10312. The purpose of this study was to compare results obtained using, on the one hand, direct preparation methods and, on the other hand, PC indirect or CE indirect methods, for counting asbestos fibres deposited on filters as a result of liquid filtration or air sampling. In direct method-based preparation, we observed that an etching time of 6-14 min does not affect the measured densities, except for fibres <1 microm deposited by liquid filtration. Moreover, in all cases, the direct method gives higher densities than the PC indirect method because of possible fibre disappearance when using the carbon evaporator implemented in the PC indirect method. The CE membrane used for sample preparation in the CE indirect method is collapsed prior to passing it through the carbon evaporator, so the fibres are less likely to disappear at this stage. We then note that the resulting fibre densities for chrysotile-loaded filters prepared using the direct method are close to those obtained with filters prepared using the CE indirect method. Our study therefore shows that, under the implemented experimental conditions, the PC and CE indirect preparation methods described in ISO Standard 13794 are not equivalent. PMID- 19812232 TI - Outcome definitions in non-dialysis intervention and prevention trials in acute kidney injury (AKI). AB - BACKGROUND: The risk, injury, failure, loss-of-function, end-stage-renal-failure (RIFLE) and acute kidney injury network (AKIN) consensus definitions of acute kidney injury (AKI) were established in part to facilitate comparison of trials. Contrast-induced nephropathy (CIN) has traditionally used a less demanding definition. OBJECTIVES: To review use of RIFLE and AKIN as AKI trial outcome variables and contrast these with outcomes for CIN. METHODS: We conducted a search of PubMed from 1 January 2005 to 31 December 2008 and 9 trial registries for randomized control trials for preventional or interventional treatment of AKI and CIN. RESULTS: RIFLE or AKIN were outcome variables in 36% (n = 8) of the published (n = 22) and 18% (n = 4) of the current (n = 22) AKI trials. RIFLE was used to triage to intervention in three trials. The urine output definition of RIFLE and AKIN was an outcome in only two trials. In 18% (n = 8) of AKI trials, the CIN definition (increase in serum creatinine of > or =25% and/or > or =44 micromol/l) was the primary outcome. This was also the primary outcome in 56% (n = 13) of published (n = 12) and current (n = 11) CIN trials. Three published CIN trials used RIFLE or AKIN as an outcome (13%). The duration over which outcomes were determined varied from 24 h to 7 days. CONCLUSIONS: Considerable heterogeneity remains in outcome variables of AKI and CIN clinical trials. Even when the RIFLE or AKIN criteria were used, they were not applied consistently. There is a need for further consensus on surrogate outcome variables. PMID- 19812233 TI - Angiotensin II receptor blocker attenuates PDGF-induced mesangial cell migration in a receptor-independent manner. AB - BACKGROUND: Clinical studies have shown that angiotensin II (Ang II) type 1 (AT1) receptor blockers (ARBs) are able to provide renoprotection independent of their blood pressure lowering effects. ARBs also are reported to suppress oxidative stress, inflammation and certain other cellular responses in a receptor independent manner. We investigated the effects of an ARB, olmesartan, on the cell migration induced by platelet-derived growth factor (PDGF), a major mitogen involved in the pathogenesis of glomerulonephritis in rat mesangial cells (RMCs). METHODS: Cell migration was determined by a modified Boyden chamber assay. The intracellular signalling pathway was examined by western blotting. AT1 receptor expression was knocked down by small interfering RNAs. The intracellular reactive oxygen species (ROS) was measured by using a fluorescent probe. The O(2)(.-) scavenging activities were studied by the electron paramagnetic resonance-spin trapping method. RESULTS: PDGF-induced cell migration was inhibited by olmesartan in AT1 receptor knockdown RMCs. Olmesartan attenuated big mitogen-activated protein (MAP) kinase 1 (BMK1) and Src activation by PDGF in AT1 receptor knockdown RMCs. PDGF-induced BMK1 activation was suppressed by the Src family tyrosine kinase inhibitors, indicating that Src exists upstream of BMK1. The NADPH oxidase inhibitors inhibited not only PDGF-induced BMK1 and Src activation but also RMC migration. The elevation in ROS generation induced by PDGF was decreased by olmesartan. Olmesartan displayed neither directly ROS scavenging activity nor the inhibition of ROS-mediated intracellular signalling in RMCs. CONCLUSIONS: Olmesartan attenuates ROS generation by PDGF, leading to the subsequent inhibition of Src/ BMK1/migration in an AT1 receptor-independent manner in RMCs. PMID- 19812234 TI - Aquaporin AQP11 in the testis: molecular identity and association with the processing of residual cytoplasm of elongated spermatids. AB - AQP11 is one of the latest aquaporin (AQP) family members found, which differs from the other AQPs by its intracellular localisation and unusual water pore nucleotides with unclear function. Despite the highest mRNA expression among organs having been reported in the testis, the testicular molecule has not been studied in detail. Immunohistochemistry of rat adult testis localised AQP11 to the elongated spermatids (ES) and no other cell types except residual bodies inside Sertoli cells. It was absent from early ES at least until stage 13, and after a first diffuse appearance in the caudal cytoplasm became concentrated in intracellular organelles by stage 17, was strongest in vesicles in the anterior cytoplasm at the final ES stages and appeared in residual bodies. Staining was detected on the distal quarter of the sperm tail only immediately before spermiation. A similar localisation was found in the mouse and developmental profiles for both the open reading frame mRNA and protein expression in 8-50 dpp testis pinpointed its first appearance coinciding with late stage ES. Sequencing of PCR products of testicular Aqp11 containing the open reading frames confirmed a full match with GenBank databases for rat, mouse and human. Western blotting revealed two or more molecular forms with the 26/27 kDa species dominating in the rat/mouse testis and the 33/34 kDa form selectively allocated to the spermatozoa. In view of intracellular vacuolation leading to polycystic kidney in Aqp11-null mice, a possible role of testicular AQP11 in the recycling of surplus cytoplasmic components of the ES and sustaining Sertoli cell capacity in the support of spermatogenesis was discussed. PMID- 19812235 TI - Locally produced relaxin may affect testis and vas deferens function in rats. AB - We have previously shown that the rat testis and vas deferens contain high levels of the relaxin receptor, RXFP1. The present study was undertaken to determine the expression of relaxin in these tissues, and the effect of exogenous relaxin on Sertoli cell proliferation and on the mRNA levels of some proteins that may contribute to epithelial secretion and tissue reorganization in the vas deferens. Relaxin mRNA levels in testis and vas deferens were much lower than in the prostate. Sertoli cells seem to be an important source of relaxin mRNA in testis. Relaxin immunoreactivity was detected in the seminiferous epithelium but not in the interstitial compartment. The relaxin precursor was expressed in the vas deferens, and relaxin immunoreactivity was detected in apical cells of the vas deferens. Castration, but not treatment with the anti-estrogen ICI 182,780, dramatically reduced relaxin mRNA levels in the prostate and vas deferens, and this effect was prevented by testosterone. Rxfp1 mRNA levels in the vas deferens and prostate were not affected by castration or treatment with ICI 182,780. Exogenous relaxin increased the incorporation of (3)H-thymidine in cultured Sertoli cells, and treatment of the vas deferens with 100 ng/ml relaxin increased the mRNA levels for the cystic fibrosis chloride channel (cystic fibrosis transmembrane regulator) about three times, and doubled mRNA levels for the inducible form of nitric oxide synthase and metalloproteinase 7. These results suggest that locally produced relaxin acts as an autocrine or paracrine agent in the testis and vas deferens to affect spermatogenesis and seminal fluid composition. PMID- 19812236 TI - Identification and characterisation of a novel GHR defect disrupting the polypyrimidine tract and resulting in GH insensitivity. AB - OBJECTIVE: GH insensitivity (GHI) is caused in the majority of cases by impaired function of the GH receptor (GHR). All but one known GHR mutation are in the coding sequence or the exon/intron boundaries. We identified and characterised the first intronic defect occurring in the polypyrimidine tract of the GHR in a patient with severe GHI. DESIGN: We investigated the effect of the novel defect on mRNA splicing using an in vitro splicing assay and a cell transfection system. METHODS: GHR was analysed by direct sequencing. To assess the effect of the novel defect, two heterologous minigenes (wild-type and mutant L1-GHR8-L2) were generated by inserting GHR exon 8 and its flanking wild-type or mutant intronic sequences into a well-characterised splicing reporter (Adml-par L1-L2). (32)P labelled pre-mRNA was generated from the two constructs and incubated in HeLa nuclear extracts or HEK293 cells. RESULTS: Sequencing of the GHR revealed a novel homozygous defect in the polypyrimidine tract of intron 7 (IVS7-6T>A). This base change does not involve the highly conserved splice site sequences, and is not predicted in silico to affect GHR mRNA splicing. Nevertheless, skipping of exon 8 from the mutant L1-GHR8-L2 mRNA was clearly demonstrated in the in vitro splicing assay and in transfected HEK293 cells. CONCLUSION: Disruption of the GHR polypyrimidine tract causes aberrant mRNA splicing leading to a mutant GHR protein. This is predicted to lack its transmembrane and intracellular domains and, thus, be incapable of transducing a GH signal. PMID- 19812237 TI - Severe XIST hypomethylation clearly distinguishes (SRY+) 46,XX-maleness from Klinefelter syndrome. AB - OBJECTIVE: 46,XX-maleness affects 1 in 20 000 live male newborns resulting in infertility and hypergonadotrophic hypogonadism. Although the phenotypes of XX males have been well described, the molecular nature of the X chromosomes remains elusive. We assessed the X inactivation status by DNA methylation analysis of four informative loci and compared those to Klinefelter syndrome (KS) and Turner syndrome. DESIGN AND METHODS: Patient cohort consisted of ten sex-determining region of the Y (SRY+) XX-males, two (SRY-) XX-males, ten 47,XXY Klinefelter men, six 45,X Turner females and ten male and female control individuals each. Methylation analysis was carried out by bisulphite sequencing of DNA from peripheral blood lymphocytes analysing X-inactive-specific transcript (XIST), phosphoglycerate kinase 1 (PGK1), ferritin, heavy peptide-like 17 (FTHL17) and short stature homeobox (SHOX). RESULTS: XIST methylation was 18% in (SRY+) XX males, and thus they were severely hypomethylated compared to (SRY-) XX-males (48%; P<0.01), Klinefelter men (44%; P<0.01) and female controls (47%; P<0.01). Turner females and male controls displayed a high degree of XIST methylation of 98 and 94% respectively. Methylation of PGK1, undergoing X inactivation, was not significantly reduced in (SRY+) XX-males compared to female controls in spite of severe XIST hypomethylation (51 vs 69%; P>0.05). FTHL17, escaping X inactivation, but undergoing cell-type-specific inactivation was similarly methylated in XX males (89%), KS patients (87%) and female controls (90%). SHOX, an X inactivation escapee located in the pseudoautosomal region, displays similarly low degrees of methylation for XX-males (7%), KS patients (7%) and female controls (9%). CONCLUSIONS: XIST hypomethylation clearly distinguishes (SRY+) XX-males from Klinefelter men. It does not, however, impair appropriate epigenetic regulation of representative X-linked loci. PMID- 19812238 TI - Synchronization dynamics in response to plaid stimuli in monkey V1. AB - Gamma synchronization has generally been associated with grouping processes in the visual system. Here, we examine in monkey V1 whether gamma oscillations play a functional role in segmenting surfaces of plaid stimuli. Local field potentials (LFPs) and spiking activity were recorded simultaneously from multiple sites in the opercular and calcarine regions while the monkeys were presented with sequences of single and superimposed components of plaid stimuli. In accord with the previous studies, responses to the single components (gratings) exhibited strong and sustained gamma-band oscillations (30-65 Hz). The superposition of the second component, however, led to profound changes in the temporal structure of the responses, characterized by a drastic reduction of gamma oscillations in the spiking activity and systematic shifts to higher frequencies in the LFP ( approximately 10% increase). Comparisons between cerebral hemispheres and across monkeys revealed robust subject-specific spectral signatures. A possible interpretation of our results may be that single gratings induce strong cooperative interactions among populations of cells that share similar response properties, whereas plaids lead to competition. Overall, our results suggest that the functional architecture of the cortex is a major determinant of the neuronal synchronization dynamics in V1. PMID- 19812239 TI - Tonic modulation of GABA release by nicotinic acetylcholine receptors in layer V of the murine prefrontal cortex. AB - By regulating the neocortical excitability, nicotinic acetylcholine receptors (nAChRs) control vigilance and cognition and are implicated in epileptogenesis. Modulation of gamma-aminobutyric acid (GABA) release often accompanies these processes. We studied how nAChRs regulate GABAergic transmission in the murine neocortex with immunocytochemical and patch-clamp methods. The cholinergic fibers densely innervated the somatosensory, visual, motor, and prefrontal cortices (PFC). Laminar distribution was broadly homogeneous, especially in the PFC. The cholinergic terminals were often adjacent to the soma and dendrites of GABAergic interneurons, but well-differentiated synapses were rare. Tonically applied nicotine (1-100 microM) increased the frequency of spontaneous GABAergic inhibitory postsynaptic currents (IPSCs) on pyramidal neurons in PFC layer V. The contribution of nAChR types was assessed by using 1 microM dihydro-beta erythroidine (DHbetaE), to block heteromeric nAChRs, and 10 nM methyllycaconitine (MLA), to block homomeric nAChRs. Both inhibitors antagonized the effect of nicotine on IPSCs, suggesting that mixed nAChR types control pyramidal neuron inhibition in layer V. To determine whether nAChRs are expressed on basket cells' terminals, we studied miniature IPSCs (mIPSCs). These were revealed using 0.5 microM tetrodotoxin and 50 microM Cd(2+) to isolate the GABAergic terminals from the action potential drive. The nicotinic stimulation of mIPSCs was antagonized by DHbetaE, but not MLA, indicating that heteromeric nAChRs prevail in GABAergic terminals. Immunocytochemistry confirmed the expression of nAChRs on basket cells' somata and terminals. Finally, when the ionotropic glutamatergic transmission was blocked, nicotine partially inhibited the IPSCs, an effect counteracted by both DHbetaE and MLA. Therefore, a fraction of nAChRs are capable of activating GABAergic interneurons that in turn inhibit other GABAergic interneurons, thereby reducing the IPSCs. We conclude that heteromeric nAChRs control GABA release presynaptically, whereas mixed nAChRs regulate both excitation and inhibition of interneurons, the balance depending on the overall glutamatergic drive. PMID- 19812240 TI - Role of late maternal thyroid hormones in cerebral cortex development: an experimental model for human prematurity. AB - Hypothyroxinemia affects 35-50% of neonates born prematurely (12% of births) and increases their risk of suffering neurodevelopmental alterations. We have developed an animal model to study the role of maternal thyroid hormones (THs) at the end of gestation on offspring's cerebral maturation. Pregnant rats were surgically thyroidectomized at embryonic day (E) 16 and infused with calcitonin and parathormone (late maternal hypothyroidism [LMH] rats). After birth, pups were nursed by normal rats. Pups born to LMH dams, thyroxine treated from E17 to postnatal day (P) 0, were also studied. In developing LMH pups, the cortical lamination was abnormal. At P40, heterotopic neurons were found in the subcortical white matter and in the hippocampal stratum oriens and alveus. The Zn positive area of the stratum oriens of hippocampal CA3 was decreased by 41.5% showing altered mossy fibers' organization. LMH pups showed delayed learning in parallel to decreased phosphorylated cAMP response element-binding protein (pCREB) and phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2) expression in the hippocampus. Thyroxine treatment of LMH dams reverted abnormalities. In conclusion, maternal THs are still essential for normal offspring's neurodevelopment even after onset of fetal thyroid function. Our data suggest that thyroxine treatment of premature neonates should be attempted to compensate for the interruption of the maternal supply. PMID- 19812241 TI - Glutamate dehydrogenase as a marker of alcohol dependence. AB - AIMS: The aim of this study was to examine glutamate dehydrogenase (GLDH) in the diagnostic combinations as a result of new findings. METHODS: GLDH, gama glutamyltransferase (GGT), aspartate-aminotranferase (AST), alanine aminotransferase (ALT) and erythrocyte mean cell volume (MCV) were assessed three times in 238 alcoholics admitted to hospital: on admission, after 24 h and after 7 days. RESULTS: All the values were significantly higher than those in healthy persons. The fastest activity decrease was seen in GLDH. The kinetics of GLDH and AST were more applicable than GGT kinetics after a week, but GLDH kinetics were most reliable. GLDH was the most specific laboratory marker with almost 90% specificity. The sensitivity of combination MCV and GLDH kinetics after 1 week of abstinence was pathognomonic by 97.2%. This decision tree gave us a model with 84.5% accuracy. CONCLUSIONS: GLDH is an equally accurate marker of alcoholism in comparison to others, if its significantly faster decrease is taken into consideration. We strongly believe that watching changes in the activity of laboratory markers of alcoholism is an effective yet overlooked aid. PMID- 19812242 TI - Patient health literacy and patient-physician information exchange during a visit. AB - BACKGROUND: Health literacy (HL), the capacity of individuals to access, understand and use health information to make informed and appropriate health related decisions, is recognized as an important concept in patient education and disease management. OBJECTIVE: To examine the relation of three levels of HL (i.e. functional, communicative and critical HL) to patient-physician information exchange during a visit. METHODS: Participants were 134 outpatients with type 2 diabetes who were under continuous care by four attending physicians at a university-affiliated hospital. The visit communication was recorded and analysed using the Roter Interaction Analysis System. Patient HL was measured through a self-reported questionnaire using newly developed self-rated scales of functional, communicative and critical HL. Sociodemographic and clinical characteristics and patient's perception of the information exchange were assessed for each patient through self-reported questionnaires and review of electronic medical records. RESULTS: Patient HL levels were related to the information exchange process during the visit. Among the three HL scales, communicative HL (the capacity to extract information, derive meaning from different forms of communication and apply new information to changing circumstances) was related to patient's perceptions of the information exchange. Further, patient communicative HL had a modifying effect on the relationship between physician's information giving and patient's perception of it, suggesting that physician's communication may be perceived differently depending on the patient's HL. CONCLUSION: The exploration of patient HL may provide a better understanding of potential barriers to patient-physician communication and patient's self-management of disease. PMID- 19812243 TI - Pseudomonas spp.-induced systemic resistance to Botrytis cinerea is associated with induction and priming of defence responses in grapevine. AB - Non-pathogenic rhizobacteria Pseudomonas spp. can reduce disease in plant tissues through induction of a defence state known as induced systemic resistance (ISR). This resistance is based on multiple bacterial determinants, but nothing is known about the mechanisms underlying rhizobacteria-induced resistance in grapevine. In this study, the ability of Pseudomonas fluorescens CHA0 and Pseudomonas aeruginosa 7NSK2 to induce resistance in grapevine against Botrytis cinerea is demonstrated. Both strains also triggered an oxidative burst and phytoalexin (i.e. resveratrol and viniferin) accumulation in grape cells and primed leaves for accelerated phytoalexin production upon challenge with B. cinerea. Treatment of cell cultures with crude cell extracts of bacteria strongly enhanced oxidative burst, but resulted in comparable amounts of phytoalexins and resistance to B. cinerea to those induced by living bacteria. This suggests the production of bacterial compounds serving as inducers of disease resistance. Using other strains with different characteristics, it is shown that P. fluorescens WCS417 (Pch-deficient), P. putida WCS358 (Pch- and SA-deficient) and P. fluorescens Q2 87 (a DAPG producer) were all capable of inducing resistance to an extent similar to that induced by CHA0. However, in response to WCS417 (Pch-negative) the amount of H2O2 induced is less than for the CHA0. WCS417 induced low phytoalexin levels in cells and lost the capacity to prime for phytoalexins in the leaves. This suggests that, depending on the strain, SA, pyochelin, and DAPG are potentially effective in inducing or priming defence responses. The 7NSK2 mutants, KMPCH (Pch and Pvd-negative) and KMPCH-567 (Pch-, Pvd-, and SA-negative) induced only partial resistance to B. cinerea. However, the amount of H2O2 triggered by KMPCH and KMPCH-567 was similar to that induced by 7NSK2. Both mutants also led to a low level of phytoalexins in grapevine cells, while KMPCH slightly primed grapevine leaves for enhanced phytoalexins. This highlights the importance of SA, pyochelin, and/or pyoverdin in priming phytoalexin responses and induced grapevine resistance by 7NSK2 against B. cinerea. PMID- 19812244 TI - Brd4 marks select genes on mitotic chromatin and directs postmitotic transcription. AB - On entry into mitosis, many transcription factors dissociate from chromatin, resulting in global transcriptional shutdown. During mitosis, some genes are marked to ensure the inheritance of their expression in the next generation of cells. The nature of mitotic gene marking, however, has been obscure. Brd4 is a double bromodomain protein that localizes to chromosomes during mitosis and is implicated in holding mitotic memory. In interphase, Brd4 interacts with P-TEFb and functions as a global transcriptional coactivator. We found that throughout mitosis, Brd4 remained bound to the transcription start sites of many M/G1 genes that are programmed to be expressed at the end of, or immediately after mitosis. In contrast, Brd4 did not bind to genes that are expressed at later phases of cell cycle. Brd4 binding to M/G1 genes increased at telophase, the end phase of mitosis, coinciding with increased acetylation of histone H3 and H4 in these genes. Increased Brd4 binding was accompanied by the recruitment of P-TEFb and de novo M/G1 gene transcription, the events impaired in Brd4 knockdown cells. In sum, Brd4 marks M/G1 genes for transcriptional memory during mitosis, and upon exiting mitosis, this mark acts as a signal for initiating their prompt transcription in daughter cells. PMID- 19812245 TI - The exomer coat complex transports Fus1p to the plasma membrane via a novel plasma membrane sorting signal in yeast. AB - Sorting of transmembrane cargo proteins to different cellular compartments is mediated by sorting signals that are recognized by coat proteins involved in vesicle biogenesis. We have identified a sorting signal in the yeast cell fusion protein Fus1p that is required for its transport from the trans-Golgi compartment to the plasma membrane. Transport of Fus1p from the trans-Golgi to the cell surface is dependent on Chs5p, a component of the multisubunit exomer complex. We show that Fus1p transport is also dependent on the exomer components Bch1p and Bud7p. Disruption of the clathrin adaptor protein complex 1 (AP-1) restores Fus1p localization to the cell surface in the absence of exomer, possibly by promoting an alternate, exomer-independent route of transport. Mutation of an IXTPK sequence in the cytosolic tail of Fus1p abolishes its physical interaction with Chs5p, results in mislocalization of Fus1p, and therefore causes a cell fusion defect. These defects are suppressed by disruption of AP-1. We suggest that IXTPK comprises a novel sorting signal that is recognized and bound by exomer leading to the capture of Fus1p into coated vesicles en route to the cell surface. PMID- 19812246 TI - Tight functional coupling of kinesin-1A and dynein motors in the bidirectional transport of neurofilaments. AB - We have tested the hypothesis that kinesin-1A (formerly KIF5A) is an anterograde motor for axonal neurofilaments. In cultured sympathetic neurons from kinesin-1A knockout mice, we observed a 75% reduction in the frequency of both anterograde and retrograde neurofilament movement. This transport defect could be rescued by kinesin-1A, and with successively decreasing efficacy by kinesin-1B and kinesin 1C. In wild-type neurons, headless mutants of kinesin-1A and kinesin-1C inhibited both anterograde and retrograde movement in a dominant-negative manner. Because dynein is thought to be the retrograde motor for axonal neurofilaments, we investigated the effect of dynein inhibition on anterograde and retrograde neurofilament transport. Disruption of dynein function by using RNA interference, dominant-negative approaches, or a function-blocking antibody also inhibited both anterograde and retrograde neurofilament movement. These data suggest that kinesin-1A is the principal but not exclusive anterograde motor for neurofilaments in these neurons, that there may be some functional redundancy among the kinesin-1 isoforms with respect to neurofilament transport, and that the activities of the anterograde and retrograde neurofilament motors are tightly coordinated. PMID- 19812247 TI - Evidence that electrostatic interactions between vesicle-associated membrane protein 2 and acidic phospholipids may modulate the fusion of transport vesicles with the plasma membrane. AB - The juxtamembrane domain of vesicle-associated membrane protein (VAMP) 2 (also known as synaptobrevin2) contains a conserved cluster of basic/hydrophobic residues that may play an important role in membrane fusion. Our measurements on peptides corresponding to this domain determine the electrostatic and hydrophobic energies by which this domain of VAMP2 could bind to the adjacent lipid bilayer in an insulin granule or other transport vesicle. Mutation of residues within the juxtamembrane domain that reduce the VAMP2 net positive charge, and thus its interaction with membranes, inhibits secretion of insulin granules in beta cells. Increasing salt concentration in permeabilized cells, which reduces electrostatic interactions, also results in an inhibition of insulin secretion. Similarly, amphipathic weak bases (e.g., sphingosine) that reverse the negative electrostatic surface potential of a bilayer reverse membrane binding of the positively charged juxtamembrane domain of a reconstituted VAMP2 protein and inhibit membrane fusion. We propose a model in which the positively charged VAMP and syntaxin juxtamembrane regions facilitate fusion by bridging the negatively charged vesicle and plasma membrane leaflets. PMID- 19812248 TI - RhoG promotes neural progenitor cell proliferation in mouse cerebral cortex. AB - In early cortical development, neural progenitor cells (NPCs) expand their population in the ventricular zone (VZ), and produce neurons. Although a series of studies have revealed the process of neurogenesis, the molecular mechanisms regulating NPC proliferation are still largely unknown. Here we found that RhoG, a member of Rho family GTPases, was expressed in the VZ at early stages of cortical development. Expression of constitutively active RhoG promoted NPC proliferation and incorporation of bromodeoxyuridine (BrdU) in vitro, and the proportion of Ki67-positive cells in vivo. In contrast, knockdown of RhoG by RNA interference suppressed the proliferation, BrdU incorporation, and the proportion of Ki67-positive cells in NPCs. However, knockdown of RhoG did not affect differentiation and survival of NPC. The RhoG-induced promotion of BrdU incorporation required phosphatidylinositol 3-kinase (PI3K) activity but not the interaction with ELMO. Taken together, these results indicate that RhoG promotes NPC proliferation through PI3K in cortical development. PMID- 19812249 TI - Dcp1-bodies in mouse oocytes. AB - Processing bodies (P-bodies) are cytoplasmic granules involved in the storage and degradation of mRNAs. In somatic cells, their formation involves miRNA-mediated mRNA silencing. Many P-body protein components are also found in germ cell granules, such as in mammalian spermatocytes. In fully grown mammalian oocytes, where changes in gene expression depend entirely on translational control, RNA granules have not as yet been characterized. Here we show the presence of P-body like foci in mouse oocytes, as revealed by the presence of Dcp1a and the colocalization of RNA-associated protein 55 (RAP55) and the DEAD box RNA helicase Rck/p54, two proteins associated with P-bodies and translational control. These P body-like structures have been called Dcp1-bodies and in meiotically arrested primary oocytes, two types can be distinguished based on their size. They also have different protein partners and sensitivities to the depletion of endogenous siRNA/miRNA and translational inhibitors. However, both type progressively disappear during in vitro meiotic maturation and are virtually absent in metaphase II-arrested secondary oocytes. Moreover, this disassembly of hDcp1a bodies is concomitant with the posttranslational modification of EGFP-hDcp1a. PMID- 19812250 TI - Rescue of Munc18-1 and -2 double knockdown reveals the essential functions of interaction between Munc18 and closed syntaxin in PC12 cells. AB - Munc18-1 binds to syntaxin-1A via two distinct sites referred to as the "closed" conformation and N terminus binding. The latter has been shown to stimulate soluble N-ethylmaleimide-sensitive factor attachment protein receptor-mediated exocytosis, whereas the former is believed to be inhibitory or dispensable. To precisely define the contributions of each binding mode, we have engineered Munc18-1/-2 double knockdown neurosecretory cells and show that not only syntaxin 1A and -1B but also syntaxin-2 and -3 are significantly reduced as a result of Munc18-1 and -2 knockdown. Syntaxin-1 was mislocalized and the regulated secretion was abolished. We next examined the abilities of Munc18-1 mutants to rescue the defective phenotypes. Mutation (K46E/E59K) of Munc18-1 that selectively prevents binding to closed syntaxin-1 was unable to restore syntaxin 1 expression, localization, or secretion. In contrast, mutations (F115E/E132A) of Munc18-1 that selectively impair binding to the syntaxin-1 N terminus could still rescue the defective phenotypes. Our results indicate that Munc18-1 and -2 act in concert to support the expression of a broad range of syntaxins and to deliver syntaxin-1 to the plasma membrane. Our studies also indicate that the binding to the closed conformation of syntaxin is essential for Munc18-1 stimulatory action, whereas the binding to syntaxin N terminus plays a more limited role in neurosecretory cells. PMID- 19812251 TI - A mutation associated with CMT2A neuropathy causes defects in Fzo1 GTP hydrolysis, ubiquitylation, and protein turnover. AB - Charcot-Marie-Tooth disease type 2A (CMT2A) is caused by mutations in the gene MFN2 and is one of the most common inherited peripheral neuropathies. Mfn2 is one of two mammalian mitofusin GTPases that promote mitochondrial fusion and maintain organelle integrity. It is not known how mitofusin mutations cause axonal degeneration and CMT2A disease. We used the conserved yeast mitofusin FZO1 to study the molecular consequences of CMT2A mutations on Fzo1 function in vivo and in vitro. One mutation (analogous to the CMT2A I213T substitution in the GTPase domain of Mfn2) not only abolishes GTP hydrolysis and mitochondrial membrane fusion but also reduces Mdm30-mediated ubiquitylation and degradation of the mutant protein. Importantly, complexes of wild type and the mutant Fzo1 protein are GTPase active and restore ubiquitylation and degradation of the latter. These studies identify diverse and unexpected effects of CMT2A mutations, including a possible role for mitofusin ubiquitylation and degradation in CMT2A pathogenesis, and provide evidence for a novel link between Fzo1 GTP hydrolysis, ubiquitylation, and mitochondrial fusion. PMID- 19812252 TI - Macrophages create an acidic extracellular hydrolytic compartment to digest aggregated lipoproteins. AB - A critical event in atherogenesis is the interaction of macrophages with subendothelial lipoproteins. Although most studies model this interaction by incubating macrophages with monomeric lipoproteins, macrophages in vivo encounter lipoproteins that are aggregated. The physical features of the lipoproteins require distinctive mechanisms for their uptake. We show that macrophages create an extracellular, acidic, hydrolytic compartment to carry out digestion of aggregated low-density lipoproteins. We demonstrate delivery of lysosomal contents to these specialized compartments and their acidification by vacuolar ATPase, enabling aggregate catabolism by lysosomal acid hydrolases. We observe transient sealing of portions of the compartments, allowing formation of an "extracellular" proton gradient. An increase in free cholesterol is observed in aggregates contained in these compartments. Thus, cholesteryl ester hydrolysis can occur extracellularly in a specialized compartment, a lysosomal synapse, during the interaction of macrophages with aggregated low-density lipoprotein. A detailed understanding of these processes is essential for developing strategies to prevent atherosclerosis. PMID- 19812253 TI - Polyamines regulate c-Myc translation through Chk2-dependent HuR phosphorylation. AB - All mammalian cells depend on polyamines for normal growth and proliferation, but the exact roles of polyamines at the molecular level remain largely unknown. The RNA-binding protein HuR modulates the stability and translation of many target mRNAs. Here, we show that in rat intestinal epithelial cells (IECs), polyamines enhanced HuR association with the 3'-untranslated region of the c-Myc mRNA by increasing HuR phosphorylation by Chk2, in turn promoting c-Myc translation. Depletion of cellular polyamines inhibited Chk2 and reduced the affinity of HuR for c-Myc mRNA; these effects were completely reversed by addition of the polyamine putrescine or by Chk2 overexpression. In cells with high content of cellular polyamines, HuR silencing or Chk2 silencing reduced c-Myc translation and c-Myc expression levels. Our findings demonstrate that polyamines regulate c Myc translation in IECs through HuR phosphorylation by Chk2 and provide new insight into the molecular functions of cellular polyamines. PMID- 19812254 TI - Neogenin regulates skeletal myofiber size and focal adhesion kinase and extracellular signal-regulated kinase activities in vivo and in vitro. AB - A variety of signaling pathways participate in the development of skeletal muscle, but the extracellular cues that regulate such pathways in myofiber formation are not well understood. Neogenin is a receptor for ligands of the netrin and repulsive guidance molecule (RGM) families involved in axon guidance. We reported previously that neogenin promoted myotube formation by C2C12 myoblasts in vitro and that the related protein Cdo (also Cdon) was a potential neogenin coreceptor in myoblasts. We report here that mice homozygous for a gene trap mutation in the Neo1 locus (encoding neogenin) develop myotomes normally but have small myofibers at embryonic day 18.5 and at 3 wk of age. Similarly, cultured myoblasts derived from such animals form smaller myotubes with fewer nuclei than myoblasts from control animals. These in vivo and in vitro defects are associated with low levels of the activated forms of focal adhesion kinase (FAK) and extracellular signal-regulated kinase (ERK), both known to be involved in myotube formation, and inefficient expression of certain muscle-specific proteins. Recombinant netrin-2 activates FAK and ERK in cultured myoblasts in a neogenin- and Cdo-dependent manner, whereas recombinant RGMc displays lesser ability to activate these kinases. Together, netrin-neogenin signaling is an important extracellular cue in regulation of myogenic differentiation and myofiber size. PMID- 19812257 TI - All-cause and cause-specific mortality of social assistance recipients in Norway: a register-based follow-up study. AB - AIMS: To investigate the mortality among social assistance recipients, who are among the most marginalized people in Norway. Cause-specific mortality was analysed in an attempt to explain the excess mortality. Previous research has suggested that social disadvantage leads to higher mortality from all causes, whereas others have found substantial variation when studying separate causes. The impact of the various causes will influence policy recommendations. METHODS: Data were compiled through linking between Norwegian administrative records. The entire population born between 1935 and 1974 (2,297,621 people) was followed with respect to social assistance and death from 1993 to 2003. Cause-specific, age standardized mortality rates for social assistance recipients and the rest of the population were calculated, and both the absolute (rate difference) and relative (rate ratio) rates were measured. RESULTS: The rate ratio for total mortality was 3.1 for men and 2.5 for women for the comparison between social assistance recipients and the general population. The mortality among social assistance recipients was higher for all causes, but the magnitude differed considerably, depending on the cause. The rate ratio for men ranged from 1.2 for non-smoking related cancer to 18.8 for alcohol- and drug-related causes. Alcohol-and drug related and violent causes together contributed to half of the excess mortality for men and one-third for women. CONCLUSIONS: The mortality of this socially disadvantaged group was considerably higher than that of the general population, and this difference reflected mainly drug-related causes. PMID- 19812256 TI - Higher C-reactive protein and soluble tumor necrosis factor receptor levels are associated with poor physical function and disability: a cross-sectional analysis of a cohort of late middle-aged African Americans. AB - BACKGROUND: This cohort of "late middle-aged" African Americans has an excess of disability. We aimed to determine associations of circulating cytokine receptors (sTNFR1, sTNFR2, and sIL-6R) and C-reactive protein (CRP) with disability, physical function, and body composition. METHODS: Stratified sampling of two socioeconomically diverse strata of St Louis, Missouri, occurred in 2000-2001. Inclusion criteria were self-reported black or African American race, born 1936 1950 inclusive, and Mini-Mental State Examination score of 16 or greater. In-home evaluations of handgrip strength, lean body mass percentage (LBM%), physical performance, upper and lower body functional limitations (UBFLs and LBFLs), and basic and instrumental activities of daily living (BADLs and IADLs) were collected. Of the 998 participants, 368 had blood sampled at baseline. Serum was stored and assayed in 2006. RESULTS: Absolute risks were LBFLs of 2 or more, 46%; UBFLs of 1 or more, 23.5%; BADLs of 2 or more, 20.6%; and IADLs of 2 or more, 22.5%. Independent of age, sex, and underlying comorbid conditions, higher CRP and sTNFR were associated with poorer physical performance (beta = -1.462, p < .001 and beta = -0.618, p = .003), UBFLs (odds ratio [OR] 2.26, 95% confidence interval [CI] 1.1-4.64 and OR 1.39, 95% CI 0.96-2.02), LBFLs (OR 2.30, 95% CI 1.19-4.45 and OR 1.91, 95% CI 1.26-2.91), BADLs (OR 2.79, 95% CI 1.03-5.96 and OR 1.66, 95% CI 1.11-2.46), and IADLs (OR 2.13, 95% CI 1.03-4.41 and OR 1.43, 95% CI 0.99-2.08). Higher CRP (beta = -3.251, p <.001), sIL-6R (beta = -6.152, p = .013), and lower adiponectin (beta = 2.947, p = .052) were associated with lower LBM%. CONCLUSIONS: Higher CRP and sTNFR are independently associated with disability and physical dysfunction. Higher sIL-6R, CRP, and lower adiponectin associate with lower LBM%. PMID- 19812255 TI - Utp9p facilitates Msn5p-mediated nuclear reexport of retrograded tRNAs in Saccharomyces cerevisiae. AB - Utp9p is a nucleolar protein that is part of a subcomplex containing several U3 snoRNA-associated proteins including Utp8p, which is a protein that shuttles aminoacyl-tRNAs from the nucleolus to the nuclear tRNA export receptors Los1p and Msn5p in Saccharomyces cerevisiae. Here we show that Utp9p is also an intranuclear component of the Msn5p-mediated nuclear tRNA export pathway. Depletion of Utp9p caused nuclear accumulation of mature tRNAs derived from intron-containing precursors, but not tRNAs made from intronless pre-tRNAs. Utp9p binds tRNA directly and saturably, and copurifies with Utp8p, Gsp1p, and Msn5p, but not with Los1p or aminoacyl-tRNA synthetases. Utp9p interacts directly with Utp8p, Gsp1p, and Msn5p in vitro. Furthermore, Gsp1p forms a complex with Msn5p and Utp9p in a tRNA-dependent manner. However, Utp9p does not shuttle between the nucleus and the cytoplasm. Because tRNA splicing occurs in the cytoplasm and the spliced tRNAs are retrograded back to the nucleus, we propose that Utp9p facilitates nuclear reexport of retrograded tRNAs. Moreover, the data suggest that Utp9p together with Utp8p translocate aminoacyl-tRNAs from the nucleolus to Msn5p and assist with formation of the Msn5p-tRNA-Gsp1p-GTP export complex. PMID- 19812259 TI - Specific serodiagnosis of canine visceral leishmaniasis using Leishmania species ribosomal protein extracts. AB - In the present work, we have analyzed the antigenicity of Leishmania species ribosomal proteins (LRPs). To accomplish this, Leishmania infantum ribosomes were biochemically purified from promastigote cytosolic extracts, and their reactivities were analyzed by using the sera from dogs naturally infected with L. infantum. Since antibodies reacting against different ribosomal proteins were observed in all the serum samples obtained from dogs with symptomatic visceral leishmaniasis tested, we have analyzed the potential usefulness of the LRP extracts in the development of an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of canine visceral leishmaniasis (CVL) in an area of Brazil where visceral leishmaniasis is endemic due to infection by Leishmania chagasi. A comparative ELISA with crude soluble Leishmania chagasi antigen (SLA) and L. infantum LRPs was performed. LRP- and SLA-based ELISAs gave similar sensitivities for the diagnosis of symptomatic CVL, but the LRP extract provided a very high sensitivity for the detection of oligosymptomatic and asymptomatic dogs. In addition, an LRP-based ELISA showed a higher specificity when the sera from dogs harboring other infections were included in the analysis. The LRP antigen displayed no cross-reactivity with sera from dogs that had any of the other diseases tested, notably, Chagas' disease. Our findings suggest that LRPs are a potential tool for the diagnosis of CVL and will be particularly useful for the diagnosis of asymptomatic CVL. PMID- 19812260 TI - Quadrivalent meningococcal vaccination of adults: phase III comparison of an investigational conjugate vaccine, MenACWY-CRM, with the licensed vaccine, Menactra. AB - Neisseria meningitidis is a leading cause of bacterial meningitis in the United States, with the highest case fatality rates reported for individuals > or = 15 years of age. This study compares the safety and immunogenicity of the Novartis Vaccines investigational quadrivalent meningococcal CRM(197) conjugate vaccine, MenACWY-CRM, to those of the licensed meningococcal conjugate vaccine, Menactra, when administered to healthy adults. In this phase III multicenter study, 1,359 adults 19 to 55 years of age were randomly assigned to one of four groups (1:1:1:1 ratio) to receive a single dose of one of three lots of MenACWY-CRM or a single dose of Menactra. Serum samples obtained at baseline and 1 month postvaccination were tested for serogroup-specific serum bactericidal activity using human complement (hSBA). The hSBA titers following vaccination with MenACWY CRM and Menactra were compared in noninferiority and prespecified superiority analyses. Reactogenicity was similar in the MenACWY-CRM and Menactra groups, and neither vaccine was associated with a serious adverse event. When compared with Menactra, MenACWY-CRM met the superiority criteria for the proportions of recipients achieving a seroresponse against serogroups C, W-135, and Y and the proportion of subjects achieving postvaccination titers of > or = 1:8 for serogroups C and Y. MenACWY-CRM's immunogenicity was statistically noninferior (the lower limit of the two-sided 95% confidence interval was more than -10%) to that of Menactra for all four serogroups, with the postvaccination hSBA geometric mean titers being consistently higher for MenACWY-CRM than for Menactra. MenACWY CRM is well tolerated in adults 19 to 55 years of age, with immune responses to each of the serogroups noninferior and, in some cases, statistically superior to those to Menactra. PMID- 19812258 TI - DNA vaccination by electroporation and boosting with recombinant proteins enhances the efficacy of DNA vaccines for Schistosomiasis japonica. AB - Schistosomiasis japonica is an endemic, zoonotic disease of major public health importance in China. Control programs combining chemotherapy and snail killing have not been able to block transmission of infection in lakes and marsh regions. Vaccination is needed as a complementary approach to the ongoing control programs. In the present study, we wanted to determine if the efficacies of DNA vaccines encoding the 23-kDa tetraspanin membrane protein (SjC23), triose phosphate isomerase (SjCTPI), and sixfold-repeated genes of the complementarity determining region 3 (CDR3) in the H chain of NP30 could be enhanced by boosting via electroporation in vivo and/or with cocktail protein vaccines. Mice vaccinated with cocktail DNA vaccines showed a significant worm reduction of 32.88% (P < 0.01) and egg reduction of 36.20% (P < 0.01). Vaccine efficacy was enhanced when animals were boosted with cocktail protein vaccines; adult worm and liver egg burdens were reduced 45.35% and 48.54%, respectively. Nearly identical results were obtained in mice boosted by electroporation in vivo, with adult worm and egg burdens reduced by 45.00% and 50.88%, respectively. The addition of a protein vaccine boost to this regimen further elevated efficacy to approximately 60% for adult worm burden and greater than 60% for liver egg reduction. The levels of interleukin-2, gamma interferon, and the ratios of immunoglobulin G2a (IgG2a)/IgG1 clearly showed that cocktail DNA vaccines induced CD4(+) Th1-type responses. Boosting via either electroporation or with recombinant proteins significantly increased associated immune responses over those seen in mice vaccinated solely with DNA vaccines. Thus, schistosome DNA vaccine efficacy was significantly enhanced via boosting by electroporation in vivo and/or cocktail protein vaccines. PMID- 19812261 TI - Biological significance of amino acid substitutions in hepatitis B surface antigen (HBsAg) for glycosylation, secretion, antigenicity and immunogenicity of HBsAg and hepatitis B virus replication. AB - Amino acid substitutions of hepatitis B surface antigen (HBsAg) may affect the antigenicity and immunogenicity of HBsAg, leading to immune escape and diagnostic failure. The amino acid positions 122 and 160 are known as determinants for HBsAg subtypes d/y and w/r, respectively. The substitution K122I has been shown to strongly affect HBsAg antigenicity. In this study, we investigated the significance of naturally occurring amino acid substitutions K122I, T123N, A159G and K160N. Both T123N and K160N substitutions resulted in additional N glycosylated forms of HBsAg, while the other mutations produced more glycosylated HBsAg compared with the wild type (wt). Detection of HBsAg by ELISA and immunofluorescence staining indicated that variant HBsAg (vtHBsAg) with K122I was not recognized by HBsAg immunoassays, while vtHBsAg with T123N, A159G, K160N and A159G/K160N had reduced antigenicity. DNA immunization in BALB/c mice revealed that wtHBsAg and vtHBsAg with T123N and K160N are able to induce antibodies to HBsAg (anti-HBs), whereas K122I and A159G greatly impair the ability of HBsAg to trigger anti-HBs responses. The cellular immune response to the HBsAg aa 29-38 epitope was enhanced by the K160N substitution. Using replication competent clones of hepatitis B virus (HBV), T123N and A159G substitutions were shown to strongly reduce virion assembly. The amino acid substitution K160N appeared to compensate for the negative effect of A159G on virion production. These results reveal complex effects of amino acid substitutions on biochemical properties of HBsAg, on antigenicity and immunogenicity, and on the replication of HBV. PMID- 19812262 TI - Effects of human papillomavirus type 16 E5 deletion mutants on epithelial morphology: functional characterization of each transmembrane domain. AB - Human papillomavirus type 16 (HPV-16) is the cause of cervical cancer. The HPV genome encodes three transforming proteins, E5, E6 and E7. E6 and E7 are the main transforming proteins of HPV, while the role of E5 is still poorly understood. Using three dimensional organotypic raft cultures we show that HaCaT human keratinocytes expressing HPV-16 E5 form a very perturbed epithelium, with simultaneous hyperkeratinization of some cells and defective differentiation of other cells. The basal layer is disturbed and many cells invade the collagen matrix. Many cells among the differentiated layers show characteristics of basal cells: progression through the cell cycle, expression of cytokeratin 14, lack of cytokeratin 1 and production of matrix metalloproteases (MMP). Using deletion mutants which encompass the three hydrophobic domains of E5, we have assigned the ability to promote invasion of the matrix to the first hydrophobic domain, and the capacity to induce MMP9 to the C-terminal four amino acids. We also show that invasion and production of MMP9 can be dissociated, as mutants that are still capable of invasion do not produce MMP9 and vice versa. PMID- 19812263 TI - Cell density-dependent increase in the level of protease-resistant prion protein in prion-infected Neuro2a mouse neuroblastoma cells. AB - Cells persistently infected with prions continuously produce protease-resistant prion protein (PrP-res). Here, we show that the PrP-res level in prion-infected Neuro2a (N2a) neuroblastoma cells decreased to 50% of their initial level over the first 48 h and then recovered by 96 h after seeding. The level of cellular prion protein (PrP(C)) also appeared to fluctuate, but did not influence the fluctuation of the PrP-res level. Prion-infected N2a cells, co-cultured with a higher number of prion-unsusceptible cells, had twice as much PrP-res as those cultured without unsusceptible cells, suggesting that cell density influences the fluctuation of PrP-res as. Direct cell-to-cell contact between cells, rather than soluble factors, was involved in the cell density-dependent increase in the PrP res level. The cholesterol content, which is known to influence PrP-res formation, also changed depending on cell density. Our data suggest that alterations in cellular microenvironments controlled by cell density influence PrP-res formation. PMID- 19812264 TI - Towards an understanding of the migration of Crimean-Congo hemorrhagic fever virus. AB - Crimean-Congo haemorrhagic fever (CCHF) is a lethal disease caused by Crimean Congo hemorrhagic fever virus (CCHFV). It is one of the most widespread medically significant tick-borne pathogens, with a distribution that coincides well with the geographical occurrence of its tick vector, Hyalomma marginatum marginatum. Sporadic outbreaks of CCHF have previously been recognized in Asia, Africa, the Middle East and Europe but, in the 21st century, outbreaks have become more frequent in former Yugoslavia, Turkey and Iran. It has been suggested that CCHFV is a migrating pathogen, but it is not clear to what extent. We have, for the first time, analysed the worldwide migration pattern of CCHFV. Our results showed that Turkey may be a donor in Europe, towards both the east and the west, while the United Arab Emirates acted as a donor in the Middle East, and China was found to be the origin for genotype 2. Finally, we showed that migration of CCHFV was unrestricted between Iran and Pakistan. Considering the distribution and coincidence of the tick vector with CCHFV and CCHF, and the fact that the tick vector is present in western Europe, future outbreaks may extend to include hitherto-naive areas, suggesting that increased surveillance and geographical mapping of this lethal pathogen are needed. PMID- 19812265 TI - Functions of Tat: the versatile protein of human immunodeficiency virus type 1. AB - Human immunodeficiency virus type 1 (HIV-1) Tat is a multifunctional protein that contributes to several pathological symptoms of HIV-1 infection as well as playing a critical role in virus replication. Tat is a robust transactivating protein that induces a variety of effects by altering the expression levels of cellular and viral genes. The functions of Tat are therefore primarily related to its role in modulation of gene expression. In this review the functions of HIV-1 Tat that have been well documented, as well as a number of novel functions that have been proposed for this protein, are discussed. Since some of the functions of Tat vary in different cell types in a concentration-dependent manner and because Tat sometimes exerts the same activity through different pathways, study of this protein has at times yielded conflicting and controversial results. Due to its pivotal role in viral replication and in disease pathogenesis, Tat and the cellular pathways targeted by Tat are potential targets for new anti-HIV drugs. PMID- 19812266 TI - Genetic relatedness of H6 subtype avian influenza viruses isolated from wild birds and domestic ducks in Korea and their pathogenicity in animals. AB - We report the genetic characterization of H6 avian influenza (AI) viruses isolated from domestic ducks and wild birds in Korea between April 2008 and April 2009. A phylogenetic analysis showed that the H6N1 viruses of wild birds and domestic ducks were of the same genotype (K-1) and were similar to the H6N1 virus isolated from a live poultry market in 2003, as six of the eight gene segments of those viruses had a common source. However, the H6N2 viruses of domestic poultry were separated into four genotypes (K-2a, K-2b, K-2c and K-2d) by at least a triple reassortment between influenza viruses of low pathogenicity from Korean poultry (H9N2 and H3N2) and viruses from aquatic birds. In an experimental infection of animals, certain H6 AI viruses replicated well in chickens and mice without pre-adaptation, indicating that H6 virus pathogenicity has the potential to be altered due to multiple reassortments, and that these reassortments could result in interspecies transmission to mammals. PMID- 19812267 TI - Regulation of Marburg virus (MARV) budding by Nedd4.1: a different WW domain of Nedd4.1 is critical for binding to MARV and Ebola virus VP40. AB - The VP40 matrix protein of Marburg virus (MARV) has been shown to be the driving force behind MARV budding, a process in which the PPPY L-domain motif of VP40 plays a critical role. Here, we report that Vps4B and Nedd4.1 play critical roles in MARV VP40-mediated budding. We showed that unidentified activities of the Nedd4.1 HECT domain, along with its E3 ubiquitin ligase activity, may be required for MARV budding. Moreover, we showed that the first WW domain of Nedd4.1, WW1, is critical for binding to MARV VP40, indicating that MARV VP40 and Ebola virus VP40 are recognized by a different WW domain of Nedd4.1. This is the first report showing that the viral L-domains containing PPxY have specificities for binding to WW domains. Our findings provide new insights into MARV budding, which may contribute to the development of novel anti-MARV therapeutic strategies. PMID- 19812268 TI - Differential activation profiles of Crimean-Congo hemorrhagic fever virus- and Dugbe virus-infected antigen-presenting cells. AB - Crimean-Congo hemorrhagic fever virus (CCHFV) is a highly pathogenic, tick-borne member of the family Bunyaviridae and the genus Nairovirus. To better elucidate the pathogenesis of CCHFV, we analysed the host innate immune response induced in antigen-presenting cells (APCs) infected in vitro by CCHFV. Monocyte-derived dendritic cells (DCs) and macrophages (MPs) were both shown to be permissive for CCHFV and to replicate the virus, as monitored by genomic and antigenomic strand quantification. Virus replication was, however, controlled, corroborating an efficient alpha interferon-induced response. The upregulation of CD-83 and CD-86 indicated that CCHFV induced a partial maturation of DCs, which were also shown to activate the secretion of interleukin (IL)-6 and IL-8, but no tumour necrosis factor alpha (TNF-alpha). On the other hand, in MPs, CCHFV infection elicited a high IL-6 and TNF-alpha response and a moderate chemokine response. Nevertheless, when we compared these APC responses with those seen after infection with Dugbe virus (DUGV), a mildly pathogenic virus genetically close to CCHFV, we found that, in spite of some similarities, DUGV induced a higher cytokine/chemokine response in MPs. These results suggest that CCHFV is able to inhibit the activation of inflammatory mediators selectively in infection in vitro and that these differences could be relevant in pathogenesis. PMID- 19812269 TI - Thogoto virus ML protein is a potent inhibitor of the interferon regulatory factor-7 transcription factor. AB - The tick-transmitted orthomyxovirus Thogoto virus (THOV) encodes the ML protein acting as a viral suppressor of the host interferon (IFN) system. Here, we describe that type I IFN is strongly induced in primary mouse embryo fibroblasts as well as plasmacytoid dendritic cells upon infection with a THOV mutant lacking the ML gene. However, wild-type THOV encoding ML suppresses induction of IFN by preventing the activation of members of the IFN regulatory factor (IRF) family. We found that reporter gene expression dependent on IRF3 and IRF7 was strongly inhibited by ML. Further experiments revealed that ML interacts with IRF7 and prevents dimerization of the transcription factor and its association with the coactivator TRAF6. Interestingly, another IRF7 activation step, nuclear translocation, is not affected by ML. Our data elucidate ML protein as a virulence factor with an IRF-specific IFN-antagonistic spectrum. PMID- 19812270 TI - Acquisition of high-level mupirocin resistance and its fitness cost among methicillin-resistant Staphylococcus aureus strains with low-level mupirocin resistance. AB - We investigated whether methicillin (meticillin)-resistant Staphylococcus aureus (MRSA) isolates with low-level mupirocin resistance can serve as recipients of a pSK41-like plasmid conferring high-level mupirocin resistance without substantial fitness cost. Our results suggest that acquisition of the plasmid conferring high level mupirocin resistance was not necessarily associated with fitness cost in some MRSA recipients with low-level mupirocin resistance. PMID- 19812271 TI - Comparison of two chromogenic media for selective isolation of vancomycin resistant enterococci from stool specimens. AB - Two chromogenic media (Chromagar VRE and chromID VRE [C-ID]) performed equally well in the direct detection of vancomycin-resistant enterococci (VRE) in stool specimens after an overnight enrichment step and a 48-h incubation period, with a sensitivity of 98.2% (56/57) for both and specificities of 96.5% (195/202) and 97.5% (197/202), respectively. However, assigning discriminatory colony color was sometimes difficult, especially on C-ID. In order to facilitate simple species identification, biochemical key reactions were implemented. PMID- 19812272 TI - Postoperative mediastinitis due to Finegoldia magna with negative blood cultures. AB - We report a case of Finegoldia magna (formerly known as Peptostreptococcus magnus) mediastinitis following coronary artery bypass in a 50-year-old patient. Even if staphylococci remain the main causative organism of postoperative mediastinitis, the responsibility of anaerobic bacteria must be considered in cases of fever and sternal drainage with negative blood cultures. PMID- 19812273 TI - Nonrandom distribution of Pseudomonas aeruginosa and Staphylococcus aureus in chronic wounds. AB - The spatial organization of Pseudomonas aeruginosa and Staphylococcus aureus in chronic wounds was investigated in the present study. Wound biopsy specimens were obtained from patients diagnosed as having chronic venous leg ulcers, and bacterial aggregates in these wounds were detected and located by the use of peptide nucleic acid-based fluorescence in situ hybridization and confocal laser scanning microscopy (CLSM). We acquired CLSM images of multiple regions in multiple sections cut from five wounds containing P. aeruginosa and five wounds containing S. aureus and measured the distance of the bacterial aggregates to the wound surface. The distance of the P. aeruginosa aggregates to the wound surface was significantly greater than that of the S. aureus aggregates, suggesting that the distribution of the bacteria in the chronic wounds was nonrandom. The results are discussed in relation to our recent finding that swab culturing techniques may underestimate the presence of P. aeruginosa in chronic wounds and in relation to the hypothesis that P. aeruginosa bacteria located in the deeper regions of chronic wounds may play an important role in keeping the wounds arrested in a stage dominated by inflammatory processes. PMID- 19812274 TI - Practical disk diffusion test for detecting group B streptococcus with reduced penicillin susceptibility. AB - Although group B streptococcus (GBS) has been considered to be uniformly susceptible to beta-lactams, the presence of GBS with reduced penicillin susceptibility (PRGBS) was recently confirmed genetically. We developed a feasible and reliable method for screening PRGBS in clinical microbiology laboratories using a combination of ceftibuten, oxacillin, and ceftizoxime disks. PMID- 19812275 TI - Population structure of Staphylococcus aureus strains isolated from intensive care unit patients in the netherlands over an 11-year period (1996 to 2006). AB - The genetic background and the presence of several virulence factors of Staphylococcus aureus isolates from intensive care unit (ICU) patients from 14 hospitals in The Netherlands isolated from 1996 until 2006 were investigated. In total, 936 methicillin-susceptible S. aureus (MSSA) and 7 methicillin-resistant S. aureus (MRSA) isolates were collected. The genetic background was determined by spa typing and multilocus sequence typing (MLST). The virulence determinants Panton-Valentine leukocidin (PVL), toxic shock syndrome toxin 1 (TSST-1), and collagen adhesion (CNA) were detected with real-time PCR assays. On the MRSA isolates, mobile resistance staphylococcal cassette chromosome mec (SCCmec) typing was performed. Among the MSSA isolates, 313 different spa types were observed. A genetic background common to MRSA clones, e.g., MLST clonal complex 1 (CC1), CC5, CC8, CC22, CC30, and CC45, was observed among 62% of the isolates. The remaining isolates were associated with MSSA-related MLST CCs. MLST CC1, CC25, and CC30 were continuously present, and other MLST CCs fluctuated over time. Two percent of the MSSA isolates harbored PVL, 21% had TSST-1, and 46% were positive for CNA. There were no changes in the prevalence of the virulence factors over time. Four MRSA isolates were typed as ST8-MRSA-IV (where ST is the MLST sequence type and IV is the SCCmec type), two were ST5-MRSA-II, and one was ST228-MRSA-I. All MRSA isolates were PVL, CNA, and TSST-1 negative except for the two ST5-MRSA-II isolates, which were TSST-1 positive. No changes in the S. aureus genetic background and the prevalence of the virulence factors PVL, CNA, and TSST 1 were observed in ICU patients in The Netherlands over time. PMID- 19812276 TI - Evaluation of hepatitis C virus core antigen assays in detecting recombinant viral antigens of various genotypes. AB - A single substitution within the hepatitis C virus core antigen sequence, A48T, which is observed in approximately 30% of individuals infected with genotype 2a virus, reduces the sensitivity of a commonly used chemiluminescence enzyme immunoassay. Quantitation of the antigen is improved by using a distinct anticore antibody with a different epitope. PMID- 19812277 TI - Evaluation of three commercial broth media for pigment detection and identification of a group B Streptococcus (Streptococcus agalactiae). AB - Detection of group B Streptococcus (GBS) strains at various bacterial concentrations was evaluated using three pigment-producing broth media. At 10(3) CFU/ml, StrepB carrot broth (SBCB), Granada instant liquid biphasic (IGLB), and Northeast Laboratory GBS screening medium (NEL-GBS) showed 100% detection, but at the lower bacterial counts, SBCB and IGLB were more sensitive than NEL-GBS after 24 h. PMID- 19812278 TI - First descriptions of blaKPC in Raoultella spp. (R. planticola and R. ornithinolytica): report from the SENTRY Antimicrobial Surveillance Program. AB - Two strains of Raoultella planticola and one of Raoultella ornithinolytica showing carbapenem resistance were recovered from patients hospitalized in New Jersey and Ohio. All patients had received previous antimicrobial treatment, including carbapenems. These strains harbored bla(KPC-2) and bla(KPC-3). Carbapenemase genes were embedded in isoforms of Tn4401 and were plasmidic and chromosomal in location. PMID- 19812279 TI - Spurious amplification of a Plasmodium vivax small-subunit RNA gene by use of primers currently used to detect P. knowlesi. AB - The PCR primers commonly used to detect Plasmodium knowlesi infections in humans were found to cross-react stochastically with P. vivax genomic DNA. A nested primer set that targets one of the P. knowlesi small-subunit rRNA genes was validated for specificity and for sensitivity of detection of <10 parasite genomes. PMID- 19812280 TI - Prevalence of ST9 methicillin-resistant Staphylococcus aureus among pigs and pig handlers in Malaysia. AB - Methicillin-resistant Staphylococcus aureus (MRSA) of sequence type 398 (ST398) has frequently been detected in pigs and pig handlers. However, in Malaysia, sampling 360 pigs and 90 pig handlers from 30 farms identified novel ST9-spa type t4358-staphylococcal cassette chromosome mec type V MRSA strains that were found to transiently colonize more than 1% of pigs and 5.5% of pig handlers. PMID- 19812281 TI - Infective endocarditis due to Citrobacter koseri in an immunocompetent adult. AB - Citrobacter koseri (formerly Citrobacter diversus) is a motile gram-negative bacillus usually arising from urinary and gastrointestinal tracts. C. koseri rarely causes infection in immunocompetent patients and, thus far, has been considered an opportunistic pathogen. We report on a 30-year-old man, with no medical past, hospitalized for infective aortic endocarditis due to C. koseri. Four weeks of antibiotherapy led to a full recovery for this patient. However, this case is unusual, as previous history and 1 year of follow-up showed no features of intercurrent immunosuppression. Microbiological diagnosis was based on using 16S rRNA gene sequencing. PMID- 19812282 TI - Deaf utopias? Reviewing the sociocultural literature on the world's "Martha's Vineyard Situations". AB - Martha's Vineyard-an island off the East Coast of the United States-is known as a community where "everyone signed" for several hundred years, a utopia in the eyes of many Deaf people. Currently, there exist around the world a number of small similar "shared signing communities," for example, in Mexico, Bali, Israel, and Ghana. A few studies about these have emerged, which give some information about the social and cultural patterns in such communities. Deaf studies researchers have begun the process of "synthesizing" and theorizing this information, and have developed typologies based on "traditional" Western urban Deaf communities. This article critically reviews the existing literature and raises new questions regarding the study and theorizing of such communities. PMID- 19812283 TI - Visual-manual exploration and posterior parietal cortex in humans. AB - Areas of human posterior parietal cortex (PPC) specialized for processing sensorimotor information associated with visually locating an object, reaching to grasp, and manually exploring that object were examined using functional MRI. Cortical activation was observed in response to three tasks: 1) saccadic eye movements, 2) visually guided reaching to grasp, and 3) manual shape discrimination. During saccadic eye movements, cortical fields within the lateral and rostral superior parietal lobe (SPL) and the caudal SPL and parieto-occipital boundary were active. During visually guided reaching to grasp, regions of cortex within the postcentral sulcus (PoCS) and rostral intraparietal sulcus (IPS) were active, as well as the caudal SPL of the left hemisphere and the medial and caudal IPS of the right hemisphere. Cortical regions at the junction of the IPS and PoCS and an area in the medial SPL were active bilaterally during shape manipulation. Only a few regions were most active during a single motor behavior, whereas several areas were highly active during two or more tasks. Hemispheric asymmetries in activation patterns were observed during visually guided reaching to grasp. The gross areal organization of human PPC is likely similar to the pattern previously described in nonhuman primates, including multifunctional regions and asymmetric processing of some manual abilities. PMID- 19812284 TI - Protein tyrosine kinase involvement in learning-produced changes in Hermissenda type B photoreceptors. AB - Learning-correlated changes in the excitability and photoresponses of Hermissenda's ocular type B photoreceptors are mediated by reductions in two distinct K(+) currents, I(A) and I(K-Ca). The suppression of these K(+) currents has been linked to conditioning-produced activation of protein kinase C (PKC). The question of whether PKC accounts completely for the changes in excitability and K(+) currents or whether other kinase(s) are involved has received little attention. In the present experiments, we asked whether protein tyrosine kinases (PTKs) might also contribute to conditioning-produced alterations in B cells. We found that the PTK inhibitors genistein and lavendustin A greatly reduced cumulative depolarization of type B cells, a short-term correlate of associative learning. This disruption occurred even when PKC activation had been either occluded by preexposure of type B cells to a phorbol ester or otherwise prevented by the pseudosubstrate inhibitor peptide PKC[19-31]. PTK inhibitors also increased the amplitude of the transient (I(A)) and delayed (I(Delayed)) components of voltage-dependent K(+) current that have previously been shown to be selectively reduced by conditioning and to contribute to cumulative depolarization. Genistein partially prevented the reduction of I(A) and I(Delayed) due to in vitro conditioning and blocked the changes in their voltage dependencies. Ionophoresis of pervanadate ion, a potent inhibitor of protein tyrosine phosphatases, depolarized type B photoreceptors and occluded conditioning-produced cumulative depolarization. Pervanadate also suppressed I(A) and I(Delayed), reduced their voltage dependence, and altered inactivation kinetics for I(A), mimicking conditioning. Western blot analysis using a phosphotyrosine antibody indicated that conditioning increased the phosphotyrosine content of many proteins within the Hermissenda CNS. Collectively, our results suggest that in addition to PKC, one or more PTKs play an important role in conditioning-produced changes in type B cell excitability. PTKs and PKCs converge to effect reductions in B cell K(+) currents during conditioning, apparently through distinct biophysical mechanisms. PMID- 19812286 TI - Been there, seen that: a neural mechanism for performing efficient visual search. AB - In everyday life, we efficiently find objects in the world by moving our gaze from one location to another. The efficiency of this process is brought about by ignoring items that are dissimilar to the target and remembering which target like items have already been examined. We trained two animals on a visual foraging task in which they had to find a reward-loaded target among five task irrelevant distractors and five potential targets. We found that both animals performed the task efficiently, ignoring the distractors and rarely examining a particular target twice. We recorded the single unit activity of 54 neurons in the lateral intraparietal area (LIP) while the animals performed the task. The responses of the neurons differentiated between targets and distractors throughout the trial. Further, the responses marked off targets that had been fixated by a reduction in activity. This reduction acted like inhibition of return in saliency map models; items that had been fixated would no longer be represented by high enough activity to draw an eye movement. This reduction could also be seen as a correlate of reward expectancy; after a target had been identified as not containing the reward the activity was reduced. Within a trial, responses to the remaining targets did not increase as they became more likely to yield a result, suggesting that only activity related to an event is updated on a moment-by-moment bases. Together, our data show that all the neural activity required to guide efficient search is present in LIP. Because LIP activity is known to correlate with saccade goal selection, we propose that LIP plays a significant role in the guidance of efficient visual search. PMID- 19812287 TI - Short-latency crossed inhibitory responses in the human soleus muscle. AB - Even though interlimb coordination is critical in bipedal locomotion, the role of muscle afferent mediated feedback is unknown. The aim of this study was to establish if ipsilateral muscle generated afferent feedback can influence contralateral muscle activation patterns in the human lower limb and to elucidate the mechanisms involved. The effect of ipsilateral tibial nerve stimulation on contralateral soleus (cSOL) responses were quantified. Three interventions were investigated, 1) electrical stimulation applied to the tibial nerve at stimulation intensities from 0 to 100% of maximal M-wave (M-max) with the cSOL contracted from 5 to 15% of maximal voluntary contraction (MVC) and 15 to 30% MVC, 2) ispsilateral tibial nerve stimulation at 75% M-max prior to, during, and following the application of ischemia to the ipsilateral thigh. 3) Electrical stimulation applied to the ipsilateral sural (SuN) and medial plantar nerves at stimulation intensities from 1 to 3 times perceptual threshold. A short-latency depression in the cSOL electromyogram (EMG; onset: 37-41 ms) was observed following ipsilateral tibial nerve stimulation. The magnitude of this depression increased (P = 0.0005 and P = 0.000001) with increasing stimulus intensities. Ischemia delayed the time of the minimum of the cSOL depression (P = 0.04). SuN and medial plantar nerve stimulation evoked a longer latency depression [average; 91.2 ms (SuN); 142 ms (medial plantar nerve)] and therefore do not contribute to the response. This is the first study to demonstrate a short-latency depression in the cSOL following ipsilateral tibial nerve stimulation. Due to its short latency, the response is spinally mediated. The involvement of crossed spinal interneurons receiving input from low-threshold muscle afferents is discussed. PMID- 19812288 TI - Dynamics of spatial distortions reveal multiple time scales of motion adaptation. AB - Prolonged exposure to consistent visual motion can significantly alter the perceived direction and speed of subsequently viewed objects. These perceptual aftereffects have provided invaluable tools with which to study the mechanisms of motion adaptation and draw inferences about the properties of underlying neural populations. Behavioral studies of the time course of motion aftereffects typically reveal a gradual process of adaptation spanning a period of multiple seconds. In contrast, neurophysiological studies have documented multiple motion adaptation effects operating over similar, or substantially faster (i.e., sub second) time scales. Here we investigated motion adaptation by measuring time dependent changes in the ability of moving stimuli to distort the perceived position of briefly presented static objects. The temporal dynamics of these motion-induced spatial distortions reveal the operation of two dissociable mechanisms of motion adaptation with differing properties. The first is rapid (subsecond), acts to limit the distortions induced by continuing motion, but is not sufficient to produce an aftereffect once the motion signal disappears. The second gradually accumulates over a period of seconds, does not modulate the size of distortions produced by continuing motion, and produces repulsive aftereffects after motion offset. These results provide new psychophysical evidence for the operation of multiple mechanisms of motion adaptation operating over distinct time scales. PMID- 19812289 TI - Noise-gated encoding of slow inputs by auditory brain stem neurons with a low threshold K+ current. AB - Phasic neurons, which do not fire repetitively to steady depolarization, are found at various stages of the auditory system. Phasic neurons are commonly described as band-pass filters because they do not respond to low-frequency inputs even when the amplitude is large. However, we show that phasic neurons can encode low-frequency inputs when noise is present. With a low-threshold potassium current (I(KLT)), a phasic neuron model responds to rising and falling phases of a subthreshold low-frequency signal with white noise. When the white noise was low-pass filtered, the phasic model also responded to the signal's trough but still not to the peak. In contrast, a tonic neuron model fired mostly to the signal's peak. To test the model predictions, whole cell slice recordings were obtained in the medial (MSO) and lateral (LSO) superior olivary neurons in gerbil from postnatal day 10 (P10) to 22. The phasic MSO neurons with strong I(KLT), mostly from gerbils aged P17 or older, showed firing patterns consistent with the preceding predictions. Moreover, injecting a virtual I(KLT) into weak-phasic MSO and tonic LSO neurons with putative weak or no I(KLT) (from gerbils younger than P17) shifted the neural response from the signal's peak to the rising phase. These findings advance our knowledge about how noise gates the signal pathway and how phasic neurons encode slow envelopes of sounds with high-frequency carriers. PMID- 19812291 TI - Computer-mouse tracking reveals TMS disruptions of prefrontal function during semantic retrieval. AB - Converging evidence from neuroimaging and neuropsychological studies is essential for understanding human frontal cortical function. We introduce a new method for studying the effects of transient disruptions of frontal activity during transcranial magnetic stimulation (TMS). Using a novel combination of TMS and computer-mouse tracking, through two experiments we tested process models of semantic competition in left ventrolateral prefrontal cortex (VLPFC). On TMS stimulation of left mid-VLPFC just after presentation of an ambiguous stimulus, participants' mouse-movement trajectories deviated more toward the incorrect target for weak associate trials than for any other trial type. This effect was extinguished when participants were simultaneously shown both target and cue stimuli. Results suggest that left mid-VLPFC is necessary to resolve semantic competition when a response is underdetermined by the stimulus and the interpretive context of the stimulus is ambiguous. Computer-mouse movements reveal the dynamics of competitive interactions as they resolve, making this technique ideally suited for studying cognitive control processes and a more sensitive index of TMS disruption than reaction time and accuracy alone. PMID- 19812292 TI - State-dependent performance of optic-flow processing interneurons. AB - Active locomotive states are metabolically expensive and require efficient sensory processing both to avoid wasteful movements and to cope with an extended bandwidth of sensory stimuli. This is particularly true for flying animals because flight, as opposed to walking or resting, imposes a steplike increase in metabolism for the precise execution and control of movements. Sensory processing itself carries a significant metabolic cost, but the principles governing the adjustment of sensory processing to different locomotor states are not well understood. We use the blowfly as a model system to study the impact on visual processing of a neuromodulator, octopamine, which is known to be involved in the regulation of flight physiology. We applied an octopamine agonist and recorded the directional motion responses of identified visual interneurons known to process self-motion-induced optic flow to directional motion stimuli. The neural response range of these neurons is increased and the response latency is reduced. We also found that, due to an elevated spontaneous spike rate, the cells' negative signaling range is increased. Meanwhile, the preferred self-motion parameters the cells encode were state independent. Our results indicate that in the blowfly energetically expensive sensory coding strategies, such as rapid, large responses, and high spontaneous spike activity could be adjusted by the neuromodulator octopamine, likely to save energy during quiet locomotor states. PMID- 19812290 TI - Quantitative analysis and biophysically realistic neural modeling of the MEG mu rhythm: rhythmogenesis and modulation of sensory-evoked responses. AB - Variations in cortical oscillations in the alpha (7-14 Hz) and beta (15-29 Hz) range have been correlated with attention, working memory, and stimulus detection. The mu rhythm recorded with magnetoencephalography (MEG) is a prominent oscillation generated by Rolandic cortex containing alpha and beta bands. Despite its prominence, the neural mechanisms regulating mu are unknown. We characterized the ongoing MEG mu rhythm from a localized source in the finger representation of primary somatosensory (SI) cortex. Subjects showed variation in the relative expression of mu-alpha or mu-beta, which were nonoverlapping for roughly 50% of their respective durations on single trials. To delineate the origins of this rhythm, a biophysically principled computational neural model of SI was developed, with distinct laminae, inhibitory and excitatory neurons, and feedforward (FF, representative of lemniscal thalamic drive) and feedback (FB, representative of higher-order cortical drive or input from nonlemniscal thalamic nuclei) inputs defined by the laminar location of their postsynaptic effects. The mu-alpha component was accurately modeled by rhythmic FF input at approximately 10-Hz. The mu-beta component was accurately modeled by the addition of approximately 10-Hz FB input that was nearly synchronous with the FF input. The relative dominance of these two frequencies depended on the delay between FF and FB drives, their relative input strengths, and stochastic changes in these variables. The model also reproduced key features of the impact of high prestimulus mu power on peaks in SI-evoked activity. For stimuli presented during high mu power, the model predicted enhancement in an initial evoked peak and decreased subsequent deflections. In agreement, the MEG-evoked responses showed an enhanced initial peak and a trend to smaller subsequent peaks. These data provide new information on the dynamics of the mu rhythm in humans and the model provides a novel mechanistic interpretation of this rhythm and its functional significance. PMID- 19812293 TI - fMRI-guided TMS on cortical eye fields: the frontal but not intraparietal eye fields regulate the coupling between visuospatial attention and eye movements. AB - It is well known that parts of a visual scene are prioritized for visual processing, depending on the current situation. How the CNS moves this focus of attention across the visual image is largely unknown, although there is substantial evidence that preparation of an action is a key factor. Our results support the view that direct corticocortical feedback connections from frontal oculomotor areas to the visual cortex are responsible for the coupling between eye movements and shifts of visuospatial attention. Functional magnetic resonance imaging (fMRI)-guided transcranial magnetic stimulation (TMS) was applied to the frontal eye fields (FEFs) and intraparietal sulcus (IPS). A single pulse was delivered 60, 30, or 0 ms before a discrimination target was presented at, or next to, the target of a saccade in preparation. Results showed that the known enhancement of discrimination performance specific to locations to which eye movements are being prepared was enhanced by early TMS on the FEF contralateral to eye movement direction, whereas TMS on the IPS resulted in a general performance increase. The current findings indicate that the FEF affects selective visual processing within the visual cortex itself through direct feedback projections. PMID- 19812294 TI - Neuronal encoding of reward value and direction of actions in the primate putamen. AB - Decision making and action selection are influenced by the values of benefit, reward, cost, and punishment. Mapping of the positive and negative values of external events and actions occurs mainly via the discharge rates of neurons in the cerebral cortex, the amygdala, and the basal ganglia. However, it remains unclear how the reward values of external events and actions encoded in the basal ganglia are integrated into reward value-based control of limb-movement actions through the corticobasal ganglia loops. To address this issue, we investigated the activities of presumed projection neurons in the putamen of macaque monkeys performing a visually instructed GO-NOGO button-press task for large and small rewards. Regression analyses of neuronal discharge rates, actions, and reward values revealed three major categories of neurons. First, neurons activated during the preinstruction delay period were selective to either the GO(large reward)-NOGO(small reward) or NOGO(large reward)-GO(small reward) combinations, although the actions to be instructed were not predictable. Second, during the postinstruction epoch, GO and NOGO action-related activities were highly selective to reward size. The pre- and postinstruction activities of a large subset of neurons were also selective to cue position or GO-response direction. Third, neurons activated during both the pre- and postinstruction epochs were selective to both action and reward size. The results support the view that putamen neurons encode reward value and direction of actions, which may be a basis for mediating the processes leading from reward-value mapping to guiding ongoing actions toward their expected outcomes and directions. PMID- 19812295 TI - Laminar and orientation-dependent characteristics of spatial nonlinearities: implications for the computational architecture of visual cortex. AB - A full understanding of the computations performed in primary visual cortex is an important yet elusive goal. Receptive field models consisting of cascades of linear filters and static nonlinearities may be adequate to account for responses to simple stimuli such as gratings and random checkerboards, but their predictions of responses to complex stimuli such as natural scenes are only approximately correct. It is unclear whether these discrepancies are limited to quantitative inaccuracies that reflect well-recognized mechanisms such as response normalization, gain controls, and cross-orientation suppression or, alternatively, imply additional qualitative features of the underlying computations. To address this question, we examined responses of V1 and V2 neurons in the monkey and area 17 neurons in the cat to two-dimensional Hermite functions (TDHs). TDHs are intermediate in complexity between traditional analytic stimuli and natural scenes and have mathematical properties that facilitate their use to test candidate models. By exploiting these properties, along with the laminar organization of V1, we identify qualitative aspects of neural computations beyond those anticipated from the above-cited model framework. Specifically, we find that V1 neurons receive signals from orientation selective mechanisms that are highly nonlinear: they are sensitive to phase correlations, not just spatial frequency content. That is, the behavior of V1 neurons departs from that of linear-nonlinear cascades with standard modulatory mechanisms in a qualitative manner: even relatively simple stimuli evoke responses that imply complex spatial nonlinearities. The presence of these findings in the input layers suggests that these nonlinearities act in a feedback fashion. PMID- 19812297 TI - Diverse vasopressin V2 receptor functionality underlying partial congenital nephrogenic diabetes insipidus. AB - X-linked congenital nephrogenic diabetes insipidus (CNDI) is characterized by a defective renal response to the antidiuretic hormone (AVP) due to variations in the arginine vasopressin receptor 2 (AVPR2) gene. In a unique group of patients, the renal insensitivity to the effects of AVP is incomplete resulting in a partial phenotype. To investigate the molecular defects, two previously published variations in the AVPR2 gene, known to cause a partial CNDI phenotype, were expressed in transiently transfected human embryonic kidney cells. One variation (p.Arg104Cys) is located in the first extracellular loop and the other variation (p.Ser329Arg) is located in the intracellular COOH terminal of the receptor protein. Western blotting showed almost equal amounts of WT-V2R and Arg104Cys-V2R protein at steady state, whereas the level of Ser329Arg-V2R protein was lower. Confocal microscopy established that WT-V2R and Arg104Cys-V2R are localized on the cellular surface while the Ser329Arg-V2R primarily accumulates within the endoplasmic reticulum resulting in reduced surface expression. Ligand binding analysis demonstrated that the B(max) for cells expressing Arg104Cys-V2R and Ser329Arg-V2R were 14.8- and 2.5-fold lower than B(max) for WT-V2R, respectively. AVP affinity (1/K(d)) for WT-V2R and the Ser329Arg-V2R was similar while 1/K(d) for Arg104Cys-V2R was increased. cAMP assay revealed that cells expressing p.Arg104Cys-V2R or p.Ser329Arg-V2R produced 1.7- and 6.8-fold lower amounts of cAMP compared with WT-V2R, respectively. In conclusion, ligand binding and signal transduction capability are dependent on localization of the amino acid variation. Striking divergences at the level of receptor functionality may thus underlie similar clinical phenotypes in CNDI. PMID- 19812296 TI - Subsecond timing in primates: comparison of interval production between human subjects and rhesus monkeys. AB - This study describes the psychometric similarities and differences in motor timing performance between 20 human subjects and three rhesus monkeys during two timing production tasks. These tasks involved tapping on a push-button to produce the same set of intervals (range of 450 to 1,000 ms), but they differed in the number of intervals produced (single vs. multiple) and the modality of the stimuli (auditory vs. visual) used to define the time intervals. The data showed that for both primate species, variability increased as a function of the length of the produced target interval across tasks, a result in accordance with the scalar property. Interestingly, the temporal performance of rhesus monkeys was equivalent to that of human subjects during both the production of single intervals and the tapping synchronization to a metronome. Overall, however, human subjects were more accurate than monkeys and showed less timing variability. This was especially true during the self-pacing phase of the multiple interval production task, a behavior that may be related to complex temporal cognition, such as speech and music execution. In addition, the well-known human bias toward auditory as opposed to visual cues for the accurate execution of time intervals was not evident in rhesus monkeys. These findings validate the rhesus monkey as an appropriate model for the study of the neural basis of time production, but also suggest that the exquisite temporal abilities of humans, which peak in speech and music performance, are not all shared with macaques. PMID- 19812298 TI - Production of the first cloned camel by somatic cell nuclear transfer. AB - In this study, we demonstrate the use of somatic cell nuclear transfer to produce the first cloned camelid, a dromedary camel (Camelus dromedarius) belonging to the family Camelidae. Donor karyoplasts were obtained from adult skin fibroblasts, cumulus cells, or fetal fibroblasts, and in vivo-matured oocytes, obtained from preovulatory follicles of superstimulated female camels by transvaginal ultrasound guided ovum pick-up, were used as cytoplasts. Reconstructed embryos were cultured in vitro for 7 days up to the hatching/hatched blastocyst stage before they were transferred to synchronized recipients on Day 6 after ovulation. Pregnancies were achieved from the embryos reconstructed from all cell types, and a healthy calf, named Injaz, was born from the pregnancy by an embryo reconstructed with cumulus cells. Genotype analyses, using 25 dromedary camel microsatellite markers, confirmed that the cloned calf was derived from the donor cell line and the ovarian tissue. In conclusion, the present study reports, for the first time, establishment of pregnancies and birth of the first cloned camelid, a dromedary camel (C. dromedarius), by use of somatic cell nuclear transfer. This has opened doors for the amelioration and preservation of genetically valuable animals like high milk producers, racing champions, and males of high genetic merit in camelids. We also demonstrated, for the first time, that adult and fetal fibroblasts can be cultured, expanded, and frozen without losing their ability to support the development of nuclear transfer embryos, a technology that may potentially be used to modify fibroblast genome by homologous recombination so as to generate genetically altered cloned animals. PMID- 19812299 TI - Progesterone inhibits oxytocin- and prostaglandin F2alpha-stimulated increases in intracellular calcium concentrations in small and large ovine luteal cells. AB - There is increasing evidence that the corpus luteum has an important role in regulating its own demise. A series of experiments was performed to study the effects of luteal concentrations of progesterone on the functions of steroidogenic luteal cells. In the first experiment, steroidogenic small luteal cells (SLCs) were separated from endothelial cells, and it was determined that it was the SLCs that contained receptors for oxytocin. Treatment with progesterone (95 muM) for as little as 1 h decreased (P < 0.05) the percentage of SLCs responding to oxytocin (10 muM) with an increase in intracellular concentrations of calcium, and this effect continued for the duration of the experiment. In a second experiment, the response to oxytocin was increased (P < 0.05) by 3 h (but not 1 h) following progesterone removal, with a further increase by 16 h. The ability of 1 muM prostaglandin F(2 alpha) (PGF(2 alpha)) to increase intracellular concentrations of calcium was also decreased (P < 0.05) by progesterone treatment. By 3 h following removal of progesterone, the percentage of steroidogenic large luteal cells (LLCs) responding to PGF(2 alpha) was increased and not different from that observed in cells 16 h after progesterone removal. Finally, cyclodextrins (methyl-beta cyclodextrin [M beta CD]) were used to remove cholesterol from the plasma membrane of luteal cells, and M beta CD loaded with cholesterol was used to put cholesterol back into the plasma membrane of progesterone-treated cells. Treatment with M beta CD reduced (P < 0.05) the responsiveness of SLCs to oxytocin and LLCs to PGF(2 alpha). Use of cholesterol loaded M beta CD returned the responsiveness of both SLCs and LLCs treated with progesterone to that observed in vehicle (no progesterone)-treated controls. In summary, intraluteal concentrations of progesterone inhibit the ability of oxytocin to increase intracellular concentrations of calcium in SLCs and the ability of PGF(2 alpha) to increase intracellular concentrations of calcium in LLCs. The highest concentration of progesterone appears to act by influencing cholesterol content of the luteal cell membranes. PMID- 19812300 TI - Expression and distribution of NADPH oxidase isoforms in human myometrium--role in angiotensin II-induced hypertrophy. AB - The renin-angiotensin system is upregulated in pregnant women and may play a role in myometrial hypertrophy during pregnancy. We examined whether angiotensin II could induce myometrial protein synthesis as determined by (3)H-leucine incorporation in an immortalized human myometrial smooth muscle cell line (ULTR cells). The effects of angiotensin II were mediated by NADPH oxidase because diphenylene iodonium abolished angiotensin II-induced protein synthesis. We investigated gene expression and cellular localization of NADPH oxidase isoforms in ULTR cells and confirmed expression of NOX1, NOX4, and NOX5 in myometrial tissue. Angiotensin II induced a cellular redistribution and upregulation of NOX5 protein without altering NOX1 and NOX4 expression. It seems the effect of angiotensin II relies on the type 1 receptor (AT1), because losartan significantly blocked angiotensin II-induced increase in (3)H-leucine incorporation. We conclude that NADPH oxidase mediates angiotensin II-stimulated protein synthesis downstream of AT1 in myometrium smooth muscle cells. PMID- 19812301 TI - SPAM1 isoforms from two tissue origins are differentially localized within ejaculated bull sperm membranes and have different roles during fertilization. AB - The deduced amino acid sequence of bull sperm, SPAM1, suggests that it possesses a transmembrane domain between the hyaluronidase and the putative zona pellucida (ZP) binding domains. The objective of this study was to determine the orientation and localization of SPAM1 in order to understand how it could fulfill these two roles. We report that two isoforms of SPAM1 are present on ejaculated bull spermatozoa: one localized on the anterior portion of the sperm head, and the other on the postacrosomal portion of the head. The first isoform is expressed intracellularly, while the second one is detected at the sperm surface with its hyaluronidase domain facing the extracellular environment. Two dimensional electrophoresis revealed that the two isoforms have different masses (80 and 70 kDa, respectively), and LC/MS/MS analyses confirmed our previously published deduced amino acid sequence of bovine SPAM1. In addition, this approach showed that the 70-kDa isoform differs from the 80-kDa isoform in its C terminus. Our results suggest that the shorter SPAM1 form originates from the epididymis, while the longer one is produced during spermatogenesis. These results clearly demonstrate that ejaculated bull sperm possess two forms of SPAM1: one (epididymal) expressed at the surface, and one (testicular) that interacts with the ZP after the acrosome exocytosis. PMID- 19812302 TI - A gradient of 2-arachidonoylglycerol regulates mouse epididymal sperm cell start up. AB - During transit through the epididymis, spermatozoa are normally kept immotile and do not attain the ability to become motile until they reach the caudal epididymis. This study was undertaken to determine whether endocannabinoids play a role in the epididymis and in particular in suppressing the ability of spermatozoa to become motile. We show that the levels of the endocannabinoid 2 arachidonoylglycerol (2-AG) are high in mouse spermatozoa isolated from the caput (head) of the epididymis, where these cells do not move (or possess sluggish and irregular motility) and decrease dramatically in spermatozoa isolated from the cauda (tail). The subsequent gradient regulates, via autocrine communication, the activity of cannabinoid receptor CNR1 (previously known as CB1) present on the sperm cell membrane and induces caudal spermatozoa to acquire the potential to become motile ("start-up"). Accordingly, the genetic or pharmacological inactivation of CNR1 increases number of motile spermatozoa in caput. Also, blockers of endocannabinoid cellular uptake inhibit the potential to move of spermatozoa and destroy the 2-AG gradient throughout the epididymis. This gradient-regulated mechanism may encourage further research for future therapies related to male infertility. PMID- 19812303 TI - An efficient method for generating transgenic mice using NaOH-treated spermatozoa. AB - Transgenic (Tg) animals are widely used in researching the characteristics of exogenous genes. Intracytoplasmic sperm injection (ICSI)-mediated transgenesis (ICSI-Tr) has been a useful method for generating Tg animals, especially in the mouse. However, the original methods using freeze-thawed spermatozoa showed severe chromosomal damage and low offspring rates after embryo transfer. Herein, we describe an improved method to generate Tg mice efficiently using a simple pretreatment of spermatozoa with 10 mM NaOH. These spermatozoa lost their plasma membrane and tail, while still maintaining nuclear integrity. Sperm heads were mixed with 0.5-5 ng/microl of the transgene for enhanced green fluorescent protein (EGFP) for 3 min to 1 h at room temperature and were then microinjected into oocytes by ICSI. The best results were obtained when treated spermatozoa were incubated with 2 ng/microl of EGFP for 10 min; 55.6% of injected embryos developed to the blastocyst stage, and more than half (56.9%) of them displayed EGFP fluorescence. Under these conditions, 12 pups of 34 offspring were positive for the transgene after transfer at the 2-cell stage into pseudopregnant recipient mice (a high rate [10.2%] from manipulated embryos). This method was found to be suitable for hybrid and inbred strains of mouse such as C57BL/6 and 129X1/Sv. Thus, a simple sperm pretreatment with NaOH before ICSI-Tr resulted in an efficient insertion of an exogenous gene into the host genome. This method allows for easy production of Tg mice, requiring fewer oocytes for micromanipulation than classical methods. PMID- 19812304 TI - mTOR signaling at a glance. PMID- 19812305 TI - Cytoskeletal dynamics in growth-cone steering. AB - Interactions between dynamic microtubules and actin filaments are essential to a wide range of cell biological processes including cell division, motility and morphogenesis. In neuronal growth cones, interactions between microtubules and actin filaments in filopodia are necessary for growth cones to make a turn. Growth-cone turning is a fundamental behaviour during axon guidance, as correct navigation of the growth cone through the embryo is required for it to locate an appropriate synaptic partner. Microtubule-actin filament interactions also occur in the transition zone and central domain of the growth cone, where actin arcs exert compressive forces to corral microtubules into the core of the growth cone and thereby facilitate microtubule bundling, a requirement for axon formation. We now have a fairly comprehensive understanding of the dynamic behaviour of the cytoskeleton in growth cones, and the stage is set for discovering the molecular machinery that enables microtubule-actin filament coupling in growth cones, as well as the intracellular signalling pathways that regulate these interactions. Furthermore, recent experiments suggest that microtubule-actin filament interactions might also be important for the formation of dendritic spines from filopodia in mature neurons. Therefore, the mechanisms coupling microtubules to actin filaments in growth-cone turning and dendritic-spine maturation might be conserved. PMID- 19812306 TI - Biogenesis of tail-anchored proteins: the beginning for the end? AB - Tail-anchored proteins are a distinct class of integral membrane proteins located in several eukaryotic organelles, where they perform a diverse range of functions. These proteins have in common the C-terminal location of their transmembrane anchor and the resulting post-translational nature of their membrane insertion, which, unlike the co-translational membrane insertion of most other proteins, is not coupled to ongoing protein synthesis. The study of tail anchored proteins has provided a paradigm for understanding the components and pathways that mediate post-translational biogenesis of membrane proteins at the endoplasmic reticulum. In this Commentary, we review recent studies that have converged at a consensus regarding the molecular mechanisms that underlie this process--namely, that multiple pathways underlie the biogenesis of tail-anchored proteins at the endoplasmic reticulum. PMID- 19812307 TI - Unravelling the ultrastructure of stress granules and associated P-bodies in human cells. AB - Stress granules are cytoplasmic ribonucleoprotein granules formed following various stresses that inhibit translation. They are thought to help protecting untranslated mRNAs until stress relief. Stress granules are frequently seen adjacent to P-bodies, which are involved in mRNA degradation and storage. We have previously shown in live cells that stress granule assembly often takes place in the vicinity of pre-existing P-bodies, suggesting that these two compartments are structurally related. Here we provide the first ultrastructural characterization of stress granules in eukaryotic cells by electron microscopy. Stress granules resulting from oxidative stress, heat-shock or protein overexpression are loosely organised fibrillo-granular aggregates of a moderate electron density, whereas P bodies are denser and fibrillar. By in situ hybridization at the electron microscopic level, we show that stress granules are enriched in poly(A)(+) mRNAs, although these represent a minor fraction of the cellular mRNAs. Finally, we show that, despite close contact with P-bodies, both domains remain structurally distinct and do not interdigitate. PMID- 19812308 TI - Force-induced cell polarisation is linked to RhoA-driven microtubule-independent focal-adhesion sliding. AB - Mechanical forces play a crucial role in controlling the integrity and functionality of cells and tissues. External forces are sensed by cells and translated into signals that induce various responses. To increase the detailed understanding of these processes, we investigated cell migration and dynamic cellular reorganisation of focal adhesions and cytoskeleton upon application of cyclic stretching forces. Of particular interest was the role of microtubules and GTPase activation in the course of mechanotransduction. We showed that focal adhesions and the actin cytoskeleton undergo dramatic reorganisation perpendicular to the direction of stretching forces even without microtubules. Rather, we found that microtubule orientation is controlled by the actin cytoskeleton. Using biochemical assays and fluorescence resonance energy transfer (FRET) measurements, we revealed that Rac1 and Cdc42 activities did not change upon stretching, whereas overall RhoA activity increased dramatically, but independently of intact microtubules. In conclusion, we demonstrated that key players in force-induced cellular reorganisation are focal-adhesion sliding, RhoA activation and the actomyosin machinery. In contrast to the importance of microtubules in migration, the force-induced cellular reorganisation, including focal-adhesion sliding, is independent of a dynamic microtubule network. Consequently, the elementary molecular mechanism of cellular reorganisation during migration is different to the one in force-induced cell reorganisation. PMID- 19812309 TI - Repression of classical nuclear export by S-nitrosylation of CRM1. AB - The karyopherin chromosomal region maintenance 1 (CRM1) is the major receptor for classical nuclear protein export. However, little is known about the regulation of CRM1 itself. Here, we report that cellular CRM1 became S-nitrosylated after extensive exposure to endogenous or exogenous nitric oxide (NO). This abrogated the interaction of CRM1 with nuclear export signals (NESs) and repressed classical protein export. Analysis by mass spectrometry and involving the use of S-nitrosylation mimetic mutations indicated that modification at either of two specific cysteines of CRM1 was sufficient to abolish the CRM1-NES association. Moreover, ectopic overexpression of the corresponding S-nitrosylation-resistant CRM1 mutants rescued NO-induced repression of nuclear export. We also found that inactivation of CRM1 by NO facilitated the nuclear accumulation of the antioxidant response transcription factor Nrf2 and transcriptional activation of Nrf2-controlled genes. Together, these data demonstrate that CRM1 is negatively regulated by S-nitrosylation under nitrosative stress. We speculate that this is important for promoting a cytoprotective transcriptional response to nitrosative stress. PMID- 19812311 TI - Alzheimer's disease affects progenitor cells through aberrant {beta}-catenin signaling. PMID- 19812310 TI - Myosin-Va-interacting protein, RILPL2, controls cell shape and neuronal morphogenesis via Rac signaling. AB - Neuronal morphology plays an essential role in neuronal function. The establishment and maintenance of neuronal morphology is intimately linked to the actin cytoskeleton; however, the molecular mechanisms that regulate changes in neuronal morphology are poorly understood. Here we identify a novel myosin-Va (MyoVa)-interacting protein, RILPL2, which regulates cellular morphology. Overexpression of this protein in young or mature hippocampal neurons results in an increase in the number of spine-like protrusions. By contrast, knockdown of endogenous RILPL2 in neurons by short hairpin RNA (shRNA) interference results in reduced spine-like protrusions, a phenotype rescued by overexpression of an shRNA insensitive RILPL2 mutant, suggesting a role for RILPL2 in both the establishment and maintenance of dendritic spines. Interestingly, we demonstrate that RILPL2 and the Rho GTPase Rac1 form a complex, and that RILPL2 is able to induce activation of Rac1 and its target, p21-activated kinase (Pak). Notably, both RILPL2-mediated morphological changes and activation of Rac1-Pak signaling were blocked by expression of a truncated tail form of MyoVa or MyoVa shRNA, demonstrating that MyoVa is crucial for proper RILPL2 function. This might represent a novel mechanism linking RILPL2, the motor protein MyoVa and Rac1 with neuronal structure and function. PMID- 19812312 TI - mTOR signaling in epileptogenesis: too much of a good thing? PMID- 19812313 TI - Neural dynamics of saccadic suppression. AB - We make fast, ballistic eye movements called saccades more often than our heart beats. Although every saccade causes a large movement of the image of the environment on our retina, we never perceive this motion. This aspect of perceptual stability is often referred to as saccadic suppression: a reduction of visual sensitivity around the time of saccades. Here, we investigated the neural basis of this perceptual phenomenon with extracellular recordings from awake, behaving monkeys in the middle temporal, medial superior temporal, ventral intraparietal, and lateral intraparietal areas. We found that, in each of these areas, the neural response to a visual stimulus changes around an eye movement. The perisaccadic response changes are qualitatively different in each of these areas, suggesting that they do not arise from a change in a common input area. Importantly, our data show that the suppression in the dorsal stream starts well before the eye movement. This clearly shows that the suppression is not just a consequence of the changes in visual input during the eye movement but rather must involve a process that actively modulates neural activity just before a saccade. PMID- 19812314 TI - Synchronous with your feelings: sensorimotor {gamma} band and empathy for pain. AB - Neuroscience studies on the social sharing of observed or imagined pain focused on whether empathic pain resonance is linked to affective or sensory nodes of the pain matrix. However, empathy, like other complex cognitive processes, is inherently linked to the activation of functional networks rather than of separate brain areas. Here, we used magnetoencephalography (MEG) to explore the relationship between empathy and functional coupling of neuronal activity in primary somatosensory (SI) and motor (MI) cortices. MEG recording was performed while healthy participants observed movie-clips depicting the static hand of a stranger model, the same hand deeply penetrated by a needle, or gently touched by a Q-tip. Subjects were asked to rate the movie-derived sensations attributed to self or to the model. For each type of clip observation, we analyzed spectral power and coherence values in alpha, beta, and gamma frequency bands. While spectral power indexes separate neural activity in SI and MI, coherence values index functional cross-talk between these two areas. No power changes of SI or MI sources were induced by observation conditions in any of the frequency bands. Crucially, gamma-band coherence values were significantly higher during needle-in hand than touch and static hand observation and correlated with self-and other referred pain ratings derived from needle-in-hand movies observation. Thus, observation of others' pain increases neuronal synchronization and cross-talk between the onlookers' sensory and motor cortices, indicating that empathic resonance relies upon the activity of functional networks more than of single areas. PMID- 19812315 TI - Plasminogen enhances neuritogenesis on laminin-1. AB - Proteins of the plasminogen activation system are broadly expressed throughout the nervous system, and key roles for these proteins in neuronal function have been demonstrated. Recent reports have established that plasminogen is synthesized in neuroendocrine tissues, making this protein and the proteolytic activity of the product of its activation, plasmin, available at sites separated anatomically from circulating, hepatocyte-derived plasminogen. Results with plasminogen-deficient humans and mice suggest a role for plasminogen in neuritogenesis. To elucidate the role of the plasminogen activation system in these processes, the function of plasminogen during neuritogenesis and neurite outgrowth was studied. It is shown here that plasminogen participates in neuritogenesis, as plasmin inhibitors reduced both neurite outgrowth and neurite length in PC-12 cells. The addition of exogenous plasminogen enhanced neurite outgrowth and neurite length in both PC-12 cells and primary cortical neurons. The proteolytic activity of plasmin was required, since mutation of the catalytic serine residue completely abolished the stimulatory activity. Furthermore, mutation of the lysine binding site within kringle 5 of the plasminogen molecule also reduced the neuritogenic activity of plasminogen. Additionally, we demonstrate that plasminogen specifically bound to laminin-1, the interaction resulted in increased plasminogen activation by tissue-type plasminogen activator, and was dependent on a functional lysine binding site within plasminogen kringle 5. Moreover, during NGF-induced neuritogenesis, laminin-1 was degraded, and this cleavage was catalyzed by plasmin. This study provides the first direct evidence that plasminogen participates in neurite outgrowth and also suggests that laminin-1 degradation by plasmin contributes to the process of neuritogenesis. PMID- 19812316 TI - The spatial patterning of mouse cone opsin expression is regulated by bone morphogenetic protein signaling through downstream effector COUP-TF nuclear receptors. AB - Cone photopigments, known as opsins, are pivotal elements and the first detection module used in color vision. In mice, cone photoreceptors are distributed throughout the retina, and short-wavelength (S) and medium-wavelength (M) opsins have unique expression patterns in the retina with a gradient along the dorsoventral axis; however, the mechanisms regulating the spatial patterning of cone opsin expression have not been well documented. The purpose of this study was to define the mechanisms regulating the spatial patterning of cone opsin expression. By analyzing knock-outs for bone morphogenetic protein (BMP) signaling, we found an essential role for BMP in forming cone opsin expression patterns in the retina; however, BMP signaling is activated only transiently in the dorsal half of the retina during early retinal development. Thus, BMP is not likely to play a direct role in opsin gene expression, which starts at a later stage of retinal development. We identified the chicken ovalbumin upstream promoter-transcription factor (COUP-TF) nuclear receptor as a link between BMP and opsin expression. BMP signaling is essential for the correct dorsoventral spatial expression of COUP-TFI and COUP-TFII. Through gain- and loss-of-function analyses, we found that both COUP-TFI and COUP-TFII are required to suppress S opsin expression in the dorsal retina but that only COUP-TFI plays an essential role in suppressing M-opsin expression in the ventral retina. Based on these findings, we propose a new molecular cascade involving BMP and COUP-TFs that conveys dorsoventral information to direct the expression of cone opsins during retinal development. PMID- 19812317 TI - Mature BDNF, but not proBDNF, reduces excitability of fast-spiking interneurons in mouse dentate gyrus. AB - Mature BDNF and its precursor proBDNF may both be secreted to exert opposite effects on synaptic plasticity in the hippocampus. However, it is unknown how proBDNF and mature BDNF affect the excitability of GABAergic interneurons and thereby regulate GABAergic inhibition. We made recordings of GABAergic spontaneous IPSCs (sIPSCs) in mouse dentate gyrus granule cells and found that chronic or acute BDNF reductions led to large increases in the sIPSC frequencies, which were TTX (tetrodotoxin) sensitive and therefore action-potential driven. Conversely, addition of mature BDNF, but not proBDNF, within minutes led to a decrease in the sIPSC frequency to 44%. Direct recordings from fast-spiking GABAergic interneurons revealed that mature BDNF reduced their excitability and depressed their action potential firing, whereas proBDNF had no effect. Using the TrkB inhibitor K-252a, or mice deficient for the common neurotrophin receptor p75(NTR), the regulation of GABAergic activity was shown specifically to be mediated by BDNF binding to the neurotrophin receptor TrkB. In agreement, immunohistochemistry demonstrated that TrkB, but not p75(NTR), was expressed in parvalbumin-positive interneurons. Our results suggest that mature BDNF decreases the excitability of GABAergic interneurons via activation of TrkB, while proBDNF does not impact on GABAergic activity. Thus, by affecting the firing of GABAergic interneurons, mature BDNF may play an important role in regulating network oscillations in the hippocampus. PMID- 19812318 TI - Modulation of neuritogenesis by a protein implicated in X-linked mental retardation. AB - Posttranscriptional regulation is an important control mechanism governing gene expression in neurons. We recently demonstrated that VCX-A, a protein implicated in X-linked mental retardation, is an RNA-binding protein that specifically binds the 5' end of capped mRNAs to prevent their decapping and decay. Previously, expression of VCX-A was reported to be testes restricted. Consistent with a role in cognitive function, we demonstrate that VCX-A is ubiquitously expressed in human tissues including the brain. Moreover, retinoic acid-induced differentiation of human SH-SY5Y neuroblastoma cells promoted the accumulation of VCX-A in distinct cytoplasmic foci within neurites that colocalize with staufen1 containing RNA granules, suggesting a role in translational suppression and/or mRNA transport. Exogenous expression of VCX-A in rat primary hippocampal neurons, which normally do not express the primate-restricted VCX proteins, promoted neurite arborization, and shRNA-directed knockdown of the VCX genes in SH-SY5Y cells resulted in a reduction of both primary and secondary neurite projections upon differentiation. We propose that the cap-binding property of VCX-A reflects a role of this protein in mRNA translational regulation. In support of this hypothesized role, we demonstrate that VCX-A can specifically bind a subset of mRNAs involved in neuritogenesis and is also capable of promoting translational silencing. Thus, VCX-A contains the capacity to modulate the stability and translation of a subset of target mRNAs involved in neuronal differentiation and arborization. It is plausible that defects of these functions in the absence of the VCX genes could contribute to a mental retardation phenotype. PMID- 19812319 TI - Chronic nicotine selectively enhances alpha4beta2* nicotinic acetylcholine receptors in the nigrostriatal dopamine pathway. AB - These electrophysiological experiments, in slices and intact animals, study the effects of in vivo chronic exposure to nicotine on functional alpha4beta2* nAChRs in the nigrostriatal dopaminergic (DA) pathway. Recordings were made in wild-type and alpha4 nicotinic acetylcholine receptor (nAChR) subunit knock-out mice. Chronic nicotine enhanced methyllycaconitine citrate hydrate-resistant, dihydro beta-erythroidine hydrobromide-sensitive nicotinic currents elicited by 3-1000 mum ACh in GABAergic neurons of the substantia nigra pars reticulata (SNr), but not in DA neurons of the substantia nigra pars compacta (SNc). This enhancement leads to higher firing rates of SNr GABAergic neurons and consequently to increased GABAergic inhibition of the SNc DA neurons. In the dorsal striatum, functional alpha4* nAChRs were not found on the neuronal somata; however, nicotine acts via alpha4beta2* nAChRs in the DA terminals to modulate glutamate release onto the medium spiny neurons. Chronic nicotine also increased the number and/or function of these alpha4beta2* nAChRs. These data suggest that in nigrostriatal DA pathway, chronic nicotine enhancement of alpha4beta2* nAChRs displays selectivity in cell type and in nAChR subtype as well as in cellular compartment. These selective events augment inhibition of SNc DA neurons by SNr GABAergic neurons and also temper the release of glutamate in the dorsal striatum. The effects may reduce the risk of excitotoxicity in SNc DA neurons and may also counteract the increased effectiveness of corticostriatal glutamatergic inputs during degeneration of the DA system. These processes may contribute to the inverse correlation between tobacco use and Parkinson's disease. PMID- 19812320 TI - Synaptic circuit abnormalities of motor-frontal layer 2/3 pyramidal neurons in an RNA interference model of methyl-CpG-binding protein 2 deficiency. AB - Rett syndrome, an autism spectrum disorder with prominent motor and cognitive features, results from mutations in the gene for methyl-CpG-binding protein 2 (MeCP2). Here, to identify cortical circuit abnormalities that are specifically associated with MeCP2 deficiency, we used glutamate uncaging and laser scanning photostimulation to survey intracortical networks in mouse brain slices containing motor-frontal cortex. We used in utero transfection of short hairpin RNA constructs to knock down MeCP2 expression in a sparsely distributed subset of layer (L) 2/3 pyramidal neurons in wild-type mice, and compared input maps recorded from transfected-untransfected pairs of neighboring neurons. The effect of MeCP2 deficiency on local excitatory input pathways was severe, with an average reduction in excitatory synaptic input from middle cortical layers (L3/5A) of >30% compared with MeCP2-replete controls. MeCP2 deficiency primarily affected the strength, rather than the topography, of excitatory intracortical pathways. Inhibitory synaptic inputs and intrinsic eletrophysiological properties were unaffected in the MeCP2-knockdown neurons. These studies indicate that MeCP2 deficiency in individual postsynaptic cortical pyramidal neurons is sufficient to induce a pathological synaptic defect in excitatory intracortical circuits. PMID- 19812321 TI - Rapid assembly of functional presynaptic boutons triggered by adhesive contacts. AB - CNS synapse assembly typically follows after stable contacts between "appropriate" axonal and dendritic membranes are made. We show that presynaptic boutons selectively form de novo following neuronal fiber adhesion to beads coated with poly-d-lysine (PDL), an artificial cationic polypeptide. As demonstrated by atomic force and live confocal microscopy, functional presynaptic boutons self-assemble as rapidly as 1 h after bead contact, and are found to contain a variety of proteins characteristic of presynaptic endings. Interestingly, presynaptic compartment assembly does not depend on the presence of a biological postsynaptic membrane surface. Rather, heparan sulfate proteoglycans, including syndecan-2, as well as others possibly adsorbed onto the bead matrix or expressed on the axon surface, are required for assembly to proceed by a mechanism dependent on the dynamic reorganization of F-actin. Our results indicate that certain (but not all) nonspecific cationic molecules like PDL, with presumably electrostatically mediated adhesive properties, can effectively bypass cognate and natural postsynaptic ligands to trigger presynaptic assembly in the absence of specific target recognition. In contrast, we find that postsynaptic compartment assembly depends on the prior presence of a mature presynaptic ending. PMID- 19812322 TI - An integrated systems analysis implicates EGR1 downregulation in simian immunodeficiency virus encephalitis-induced neural dysfunction. AB - Human immunodeficiency virus (HIV)-associated dementia (HAD) is a syndrome occurring in HIV-infected patients with advanced disease that likely develops as a result of macrophage and microglial activation as well as other immune events triggered by virus in the central nervous system. The most relevant experimental model of HAD, rhesus macaques exhibiting simian immunodeficiency virus (SIV) encephalitis (SIVE), closely reproduces the human disease and has been successfully used to advance our understanding of mechanisms underlying HAD. In this study we integrate gene expression data from uninfected and SIV-infected hippocampus with a human protein interaction network and discover modules of genes whose expression patterns distinguish these two states, to facilitate identification of neuronal genes that may contribute to SIVE/HIV cognitive deficits. Using this approach we identify several downregulated candidate genes and select one, EGR1, a key molecule in hippocampus-related learning and memory, for further study. We show that EGR1 is downregulated in SIV-infected hippocampus and that it can be downregulated in differentiated human neuroblastoma cells by treatment with CCL8, a product of activated microglia. Integration of expression data with protein interaction data to discover discriminatory modules of interacting proteins can be usefully used to prioritize differentially expressed genes for further study. Investigation of EGR1, selected in this manner, indicates that its downregulation in SIVE may occur as a consequence of the host response to infection, leading to deficits in cognition. PMID- 19812323 TI - Vestibular asymmetry as the cause of idiopathic scoliosis: a possible answer from Xenopus. AB - Human idiopathic scoliosis is characterized by severe deformations of the spine and skeleton. The occurrence of vestibular-related deficits in these patients is well established but it is unclear whether a vestibular pathology is the common cause for the scoliotic syndrome and the gaze/posture deficits or if the latter behavioral deficits are a consequence of the scoliotic deformations. A possible vestibular origin was tested in the frog Xenopus laevis by unilateral removal of the labyrinthine endorgans at larval stages. After metamorphosis into young adult frogs, X-ray images and three-dimensional reconstructed micro-computer tomographic scans of the skeleton showed deformations similar to those of scoliotic patients. The skeletal distortions consisted of a curvature of the spine in the frontal and sagittal plane, a transverse rotation along the body axis and substantial deformations of all vertebrae. In terrestrial vertebrates, the initial postural syndrome after unilateral labyrinthectomy recovers over time and requires body weight-supporting limb proprioceptive information. In an aquatic environment, however, this information is absent. Hence, the lesion induced asymmetric activity in descending spinal pathways and the resulting asymmetric muscular tonus persists. As a consequence the mostly cartilaginous skeleton of the frog tadpoles progressively deforms. Lack of limb proprioceptive signals in an aquatic environment is thus the element, which links the Xenopus model with human scoliosis because a comparable situation occurs during gestation in utero. A permanently imbalanced activity in descending locomotor/posture control pathways might be the common origin for the observed structural and behavioral deficits in humans as in the different animal models of scoliosis. PMID- 19812324 TI - Chronic exposure to anabolic androgenic steroids alters neuronal function in the mammalian forebrain via androgen receptor- and estrogen receptor-mediated mechanisms. AB - Anabolic androgenic steroids (AAS) can promote detrimental effects on social behaviors for which GABA type A (GABA(A)) receptor-mediated circuits in the forebrain play a critical role. While all AAS bind to androgen receptors (AR), they may also be aromatized to estrogens and thus potentially impart effects via estrogen receptors (ER). Chronic exposure of wild-type male mice to a combination of chemically distinct AAS increased action potential (AP) frequency, selective GABA(A) receptor subunit mRNAs, and GABAergic synaptic current decay in the medial preoptic area (mPOA). Experiments performed with pharmacological agents and in AR-deficient Tfm mutant mice suggest that the AAS-dependent enhancement of GABAergic transmission in wild-type mice is AR-mediated. In AR-deficient mice, the AAS elicited dramatically different effects, decreasing AP frequency, spontaneous IPSC amplitude and frequency and the expression of selective GABA(A) receptor subunit mRNAs. Surprisingly, in the absence of AR signaling, the data indicate that the AAS do not act as ER agonists, but rather suggest a novel in vivo action in which the AAS inhibit aromatase and impair endogenous ER signaling. These results show that the AAS have the capacity to alter neuronal function in the forebrain via multiple steroid signaling mechanisms and suggest that effects of these steroids in the brain will depend not only on the balance of AR- versus ER-mediated regulation for different target genes, but also on the ability of these drugs to alter steroid metabolism and thus the endogenous steroid milieu. PMID- 19812325 TI - Essential role of cytoplasmic cdk5 and Prx2 in multiple ischemic injury models, in vivo. AB - Recent evidence suggests that abnormal activation of cyclin-dependent kinase 5 (cdk5) is a critical prodeath signal in stroke. However, the mechanism(s) by which cdk5 promotes death is unclear. Complicating the role of cdk5 are the observations that cdk5 can exist in multiple cellular regions and possess both prosurvival and prodeath characteristics. In particular, the critical role of cytoplasmic or nuclear cdk5 in neuronal jury, in vivo, is unclear. Therefore, we determined where cdk5 was activated in models of ischemia and how manipulation of cdk5 in differing compartments may affect neuronal death. Here, we show a critical function for cytoplasmic cdk5 in both focal and global models of stroke, in vivo. Cdk5 is activated in the cytoplasm and expression of DNcdk5 localized to the cytoplasm is protective. Importantly, we also demonstrate the antioxidant enzyme Prx2 (peroxiredoxin 2) as a critical cytoplasmic target of cdk5. In contrast, the role of cdk5 in the nucleus is context-dependent. Following focal ischemia, nuclear cdk5 is activated and functionally relevant while there is no evidence for such activation following global ischemia. Importantly, myocyte enhancer factor 2D (MEF2D), a previously described nuclear target of cdk5 in vitro, is also phosphorylated by cdk5 following focal ischemia. In addition, MEF2D expression in this paradigm ameliorates death. Together, our results address the critical issue of cdk5 activity compartmentalization, as well as define critical substrates for both cytoplasmic and nuclear cdk5 activity in adult models of stroke. PMID- 19812326 TI - Sonic hedgehog regulates integrin activity, cadherin contacts, and cell polarity to orchestrate neural tube morphogenesis. AB - In vertebrates, the embryonic nervous system is shaped and patterned by a series of temporally and spatially regulated cell divisions, cell specifications, and cell adhesions and movements. Morphogens of the Hedgehog, Wnt, and bone morphogenetic protein families have been shown to play a crucial role in the control of cell division and specification in the trunk neural tube, but their possible implication in the regulation of adhesive events has been poorly documented. In the present study, we demonstrate that Sonic hedgehog regulates neural epithelial cell adhesion and polarity through regulation of integrin activity, cadherin cell-cell contact, and cell polarity genes in immature neural epithelial cells before the specification of neuronal cells. We propose that Sonic hedgehog orchestrates neural tube morphogenesis by coordinating adhesive and motility events with cell proliferation and differentiation. PMID- 19812327 TI - Gamma-band synchronization in the macaque hippocampus and memory formation. AB - Increasing evidence suggests that neuronal synchronization in the gamma band (30 100 Hz) may play an important role in mediating cognitive processes. Gamma-band synchronization provides for the optimal temporal relationship between two signals to produce the long-term synaptic changes that have been theorized to underlie memory formation. Although neuronal populations in the hippocampus oscillate in the gamma range, the role of these oscillations in memory formation is still unclear. To address this issue, we recorded neuronal activity in the hippocampus while macaque monkeys performed a visual recognition memory task. During the encoding phase of this task, hippocampal neurons displayed gamma-band synchronization. Additionally, enhanced gamma-band synchronization during encoding predicted greater subsequent recognition memory performance. These changes in synchronization reflect enhanced coordination among hippocampal neurons and may facilitate synaptic changes necessary for successful memory encoding. PMID- 19812328 TI - Neurons of the dopaminergic/calcitonin gene-related peptide A11 cell group modulate neuronal firing in the trigeminocervical complex: an electrophysiological and immunohistochemical study. AB - Activation of spinal trigeminal afferents innervating the cranial vasculature is likely to play a role in migraine, although some parts of the clinical presentation may have a dopaminergic basis. The A11 nucleus, located in the posterior hypothalamus, provides the only known source of descending dopaminergic innervation for the spinal gray matter. Extracellular recordings were made in the trigeminocervical complex (TCC) in response to electrical stimulation of the dura mater. Receptive fields were characterized by mechanical noxious and innocuous stimulation of the ipsilateral ophthalmic dermatome. Stimulation of the A11 significantly inhibited peri-middle meningeal artery dural and noxious pinch evoked firing of neurons in the TCC. This inhibition was reversed by the D(2) receptor antagonist eticlopride. Lesioning of the A11 significantly facilitated dural and noxious pinch and innocuous brush evoked firing from the TCC. In previous work using immunohistofluorescence, it was shown that D(1) and D(2) receptors were found in the rat TCC, and here we report, in addition, that D(4) and D(5) dopamine receptors are also present, whereas D(3) receptors are not. No dopamine receptors were present in the A11 nucleus itself. However, the A11 does contain dopamine and calcitonin gene-related peptide (CGRP) and, by this combination, is distinct from the neighboring CGRPergic subparafascicular nucleus. Exploration of dopaminergic influences and mechanisms in migraine may open up an almost untapped opportunity to pursue potential new therapeutic options for the disorder. PMID- 19812329 TI - Semaphorin 3F is a bifunctional guidance cue for dopaminergic axons and controls their fasciculation, channeling, rostral growth, and intracortical targeting. AB - Dopaminergic neurons in the mesodiencephalon (mdDA neurons) make precise synaptic connections with targets in the forebrain via the mesostriatal, mesolimbic, and mesoprefrontal pathways. Because of the functional importance of these remarkably complex ascending axon pathways and their implication in human disease, the mechanisms underlying the development of these connections are of considerable interest. Despite extensive in vitro studies, the molecular determinants that ensure the perfect formation of these pathways in vivo remain mostly unknown. Here, we determine the embryonic origin and ontogeny of the mouse mesoprefrontal pathway and use these data to reveal an unexpected requirement for semaphorin 3F (Sema3F) and its receptor neuropilin-2 (Npn-2) during mdDA pathway development using tissue culture approaches and analysis of sema3F(-/-), npn-2(-/-), and npn 2(-/-);TH-Cre mice. We show that Sema3F is a bifunctional guidance cue for mdDA axons, some of which have the remarkable ability to regulate their responsiveness to Sema3F as they develop. During early developmental stages, Sema3F chemorepulsion controls previously uncharacterized aspects of mdDA pathway development through both Npn-2-dependent (axon fasciculation and channeling) and Npn-2-independent (rostral growth) mechanisms. Later on, chemoattraction mediated by Sema3F and Npn-2 is required to orient mdDA axon projections in the cortical plate of the medial prefrontal cortex. This latter finding demonstrates that regulation of axon orientation in the target field occurs by chemoattractive mechanisms, and this is likely to also apply to other neural systems. In all, this study provides a framework for additional dissection of the molecular basis of mdDA pathway development and disease. PMID- 19812330 TI - The maturation of task set-related activation supports late developmental improvements in inhibitory control. AB - The ability to voluntarily inhibit a single response is evident early in development, even as the ability to maintain an inhibitory "task set" continues to improve. To date, functional neuroimaging studies have detailed developmental changes in systems supporting inhibitory control exerted at the single-trial level, but changes underlying the ability to maintain an inhibitory task set remain little understood. Here we present findings from a functional magnetic resonance imaging study that characterizes the development of systems supporting both transient (trial-related) and sustained (task set-related) activation during performance of the antisaccade task-an oculomotor test of inhibitory control (Hallett, 1978). Transient activation decreased from childhood to adolescence in regions known to support inhibitory processes and oculomotor control, likely reflecting less effortful response production. In contrast, sustained activation increased to adulthood in regions implicated in control. Our results suggest that development of the ability to maintain a task set is primary to the maturation of inhibitory control and, furthermore, that this ability is still immature in adolescence. PMID- 19812331 TI - Temporal order processing of syllables in the left parietal lobe. AB - Speech processing requires the temporal parsing of syllable order. Individuals suffering from posterior left hemisphere brain injury often exhibit temporal processing deficits as well as language deficits. Although the right posterior inferior parietal lobe has been implicated in temporal order judgments (TOJs) of visual information, there is limited evidence to support the role of the left inferior parietal lobe (IPL) in processing syllable order. The purpose of this study was to examine whether the left inferior parietal lobe is recruited during temporal order judgments of speech stimuli. Functional magnetic resonance imaging data were collected on 14 normal participants while they completed the following forced-choice tasks: (1) syllable order of multisyllabic pseudowords, (2) syllable identification of single syllables, and (3) gender identification of both multisyllabic and monosyllabic speech stimuli. Results revealed increased neural recruitment in the left inferior parietal lobe when participants made judgments about syllable order compared with both syllable identification and gender identification. These findings suggest that the left inferior parietal lobe plays an important role in processing syllable order and support the hypothesized role of this region as an interface between auditory speech and the articulatory code. Furthermore, a breakdown in this interface may explain some components of the speech deficits observed after posterior damage to the left hemisphere. PMID- 19812332 TI - Neural correlates of value, risk, and risk aversion contributing to decision making under risk. AB - Decision making under risk is central to human behavior. Economic decision theory suggests that value, risk, and risk aversion influence choice behavior. Although previous studies identified neural correlates of decision parameters, the contribution of these correlates to actual choices is unknown. In two different experiments, participants chose between risky and safe options. We identified discrete blood oxygen level-dependent (BOLD) correlates of value and risk in the ventral striatum and anterior cingulate, respectively. Notably, increasing inferior frontal gyrus activity to low risk and safe options correlated with higher risk aversion. Importantly, the combination of these BOLD responses effectively decoded the behavioral choice. Striatal value and cingulate risk responses increased the probability of a risky choice, whereas inferior frontal gyrus responses showed the inverse relationship. These findings suggest that the BOLD correlates of decision factors are appropriate for an ideal observer to detect behavioral choices. More generally, these biological data contribute to the validity of the theoretical decision parameters for actual decisions under risk. PMID- 19812333 TI - A protein interaction node at the neurotransmitter release site: domains of Aczonin/Piccolo, Bassoon, CAST, and rim converge on the N-terminal domain of Munc13-1. AB - Multidomain scaffolding proteins organize the molecular machinery of neurotransmitter vesicle dynamics during synaptogenesis and synaptic activity. We find that domains of five active zone proteins converge on an interaction node that centers on the N-terminal region of Munc13-1 and includes the zinc-finger domain of Rim1, the C-terminal region of Bassoon, a segment of CAST1/ELKS2, and the third coiled-coil domain (CC3) of either Aczonin/Piccolo or Bassoon. This multidomain complex may constitute a center for the physical and functional integration of the protein machinery at the active zone. An additional connection between Aczonin and Bassoon is mediated by the second coiled-coil domain of Aczonin. Recombinant Aczonin-CC3, expressed in cultured neurons as a green fluorescent protein fusion protein, is targeted to synapses and suppresses vesicle turnover, suggesting involvements in synaptic assembly as well as activity. Our findings show that Aczonin, Bassoon, CAST1, Munc13, and Rim are closely and multiply interconnected, they indicate that Aczonin-CC3 can actively participate in neurotransmitter vesicle dynamics, and they highlight the N terminal region of Munc13-1 as a hub of protein interactions by adding three new binding partners to its mechanistic potential in the control of synaptic vesicle priming. PMID- 19812334 TI - Alteration of theta timescale dynamics of hippocampal place cells by a cannabinoid is associated with memory impairment. AB - The integrity of the hippocampus is critical for both spatial navigation and episodic memory, but how its neuronal firing patterns underlie those functions is not well understood. In particular, the modality by which hippocampal place cells contribute to spatial memory is debated. We found that administration of the cannabinoid receptor agonist CP55940 (2-[(1S,2R,5S)-5-hydroxy-2-(3 hydroxypropyl)cyclohexyl]-5-(2-methyloctan-2-yl)phenol) induced a profound and reversible behavioral deficit in the hippocampus-dependent delayed spatial alternation task. On the one hand, despite severe memory impairment, the location dependent firing of CA1 hippocampal place cells remained mostly intact. On the other hand, both spike-timing coordination between place cells at the theta timescale and theta phase precession of spikes were reversibly reduced. These results raise the possibility that cannabinoids impair memory primarily by altering short-term temporal dynamics of hippocampal neurons. We hypothesize that precise temporal coordination of hippocampal neurons is necessary for guiding behavior in spatial memory tasks. PMID- 19812335 TI - Impedance control reduces instability that arises from motor noise. AB - There is ample evidence that humans are able to control the endpoint impedance of their arms in response to active destabilizing force fields. However, such fields are uncommon in daily life. Here, we examine whether the CNS selectively controls the endpoint impedance of the arm in the absence of active force fields but in the presence of instability arising from task geometry and signal-dependent noise (SDN) in the neuromuscular system. Subjects were required to generate forces, in two orthogonal directions, onto four differently curved rigid objects simulated by a robotic manipulandum. The endpoint stiffness of the limb was estimated for each object curvature. With increasing curvature, the endpoint stiffness increased mainly parallel to the object surface and to a lesser extent in the orthogonal direction. Therefore, the orientation of the stiffness ellipses did not orient to the direction of instability. Simulations showed that the observed stiffness geometries and their pattern of change with instability are the result of a tradeoff between maximizing the mechanical stability and minimizing the destabilizing effects of SDN. Therefore, it would have been suboptimal to align the stiffness ellipse in the direction of instability. The time course of the changes in stiffness geometry suggests that modulation takes place both within and across trials. Our results show that an increase in stiffness relative to the increase in noise can be sufficient to reduce kinematic variability, thereby allowing stiffness control to improve stability in natural tasks. PMID- 19812336 TI - Effects of Lorazepam and citalopram on human defensive reactions: ethopharmacological differentiation of fear and anxiety. AB - Drugs that are clinically effective against generalized anxiety disorder preferentially alter rodent risk assessment behavior, whereas drugs that are clinically effective against panic disorder preferentially alter rodent flight behavior. The theoretical principle of "defensive direction" explains the pattern of associations between emotion and defensive behavior in terms of the differing functional demands arising from cautious approach to threat (anxiety) versus departure from threat (fear), offering the prospect that clinically important emotions may be explained using a single rubric of defense. We used a within subjects, placebo-controlled, design to test this theory, measuring the effects of citalopram and lorazepam on the defensive behavior of 30 healthy adult male humans. We indexed human defensive behavior with a translation of an active avoidance task used to measure rodent defense and found that lorazepam significantly reduced the intensity of defensive behavior during approach to threat (hypothetically anxiety-related) but not departure from threat (hypothetically fear-related). Contrary to prediction, citalopram did not affect either form of defensive reaction. Since lorazepam is a drug with well established anxiety reducing properties, these data support the hypothesis that anxiety is an emotion elicited by threat stimuli that require approach. These data also contribute to the validation of a novel human analog of an established experimental model of rodent fear and anxiety. PMID- 19812337 TI - Mouse RIC-3, an endoplasmic reticulum chaperone, promotes assembly of the alpha7 acetylcholine receptor through a cytoplasmic coiled-coil domain. AB - RIC-3 (resistant to inhibitor of cholinesterase) is a transmembrane protein, found in invertebrates and vertebrates, that modulates the surface expression of a variety of nicotinic acetylcholine receptors (nAChRs) in neurons and other cells. To understand its mechanism of action, we investigated the cellular location, transmembrane topology and cellular mechanism by which RIC-3 facilitates alpha7 assembly and surface expression in cultured mammalian cells. We show that the mouse protein is targeted to the ER by the first 31 aa which act as a cleavable signal sequence. The mature protein is a single-pass type I transmembrane protein whose N terminus resides in the lumen of the ER with the coiled-coil domain in the cytoplasm. RIC-3, which binds both unfolded and folded alpha7 subunits, facilitates the surface expression of receptor principally by promoting the folding and assembly of the alpha7 subunits in the ER into fully polymerized receptor. Functional analysis shows that facilitation of surface expression of alpha7 in mammalian cells is reduced in RIC-3 mutants lacking the signal peptide, the lumenal segment or the coiled-coil domain, but not in mutants lacking the long C-terminal region downstream of the coiled-coil domain. We show that the coiled-coil domain of mRIC-3 is not required for the interaction of mRIC 3 with alpha7, but does mediate a homotypic interaction between molecules of mRIC 3. We suggest that efficient assembly of the homomeric alpha7 nAChR may thus require mRIC-3 self-association through the cytoplasmic coiled-coil domain and suggest a model by which this may occur. PMID- 19812338 TI - A reporter of local dendritic translocation shows plaque- related loss of neural system function in APP-transgenic mice. AB - Although neuronal communication is thought to be summated within local dendritic segments, no technique is currently available to monitor activity in vivo at this level of resolution. To overcome this challenge, we developed an optical reporter of neuronal activity using the coding sequence of Venus, flanked by short stretches of the 5'- and 3'-untranslated regions from calcium/calmodulin dependent kinase IIalpha (CAMKIIalpha). This reporter takes advantage of the fact that CAMKIIalpha mRNA is transported to the dendrite and locally translated in an activity-dependent manner. Using adeno-associated virus, we used this reporter to study neuronal activity in adult mice. Exposure of the mice to an enriched environment led to enhancement of Venus expression in dendritic segments of somatosensory cortex, demonstrating in vivo that dendritic mRNA translocation and local translation occur in response to physiologically relevant stimuli. We then used this system to examine the impact of Alzheimer-related local amyloid-beta deposits on neural system function to test the hypothesis that plaques are toxic. In APPswe/PS1dE9 (APP/PS1) mice, neurons close to plaques, and dendritic segments close to plaques, both showed diminished fluorescent intensity and therefore neuronal activity. In contrast to wild-type mice, fluorescent intensity in neurons near plaques in transgenic mice did not increase after environmental enrichment. These data indicate that neuronal activity in dendritic segments and neurons in the vicinity of a plaque is decreased compared with wild-type mice, supporting the idea that plaques are a focal lesion leading to impaired neural system function. PMID- 19812339 TI - Properties of piriform cortex pyramidal cell dendrites: implications for olfactory circuit design. AB - Unlike the neocortex, sensory input to the piriform cortex is anatomically segregated in layer 1, making it ideal for studying the dendritic integration of synaptic inputs pivotal for sensory information processing. Here we investigated dendritic integration of olfactory bulb inputs in pyramidal cells using dual patch-clamp recordings along the soma-apical dendritic axis. We found that these dendrites are relatively compact with 50% maximal somatic current loss for synaptic inputs arriving at distal dendritic regions. Distal dendrites could generate small and fast local spikes, but they had little impact on the soma, indicating that they are only weakly active. In contrast to the neocortex, we found no evidence for dendritic Ca(2+) or NMDA spikes though these dendrites actively supported action potential backpropagation with concomitant entry of Ca(2+) ions. Based on experiments and simulations we suggest that regardless of dendritic location, olfactory bulb inputs have nearly uniform potency and are distributed diffusely over the distal apical tree (layer Ia), thereby minimizing sublinear summation effects. This indicates that any stimulus feature extraction performed by these cells will occur at the soma and is based on the nearly linear sum of olfactory bulb inputs, rather than on explicitly designed clusters of functionally related synapses in the dendritic tree. PMID- 19812340 TI - Long-term modifications in motor cortical dynamics induced by intensive practice. AB - The planning of goal-directed movements requires sensory, temporal, and contextual information to be combined. Sensorimotor functions are embedded in large neuronal networks, but it is unclear how networks organize their activity in space and time to optimize behavior. Temporal coordination of activity in many neurons within a network, e.g., spike synchrony, might be complementary to a firing rate code, allowing efficient computation with overall less population activity. Here we asked the question whether intensive practice induces long-term modifications in the temporal structure of synchrony and firing rate at the population level. Three monkeys were trained in a delayed pointing task in which the selection of movement direction depended on correct time estimation. The synchronous firing among pairs of simultaneously recorded neurons in motor cortex was analyzed using the "unitary event" technique. The evolution of synchrony in both time, within the trial, and temporal precision was then quantified at the level of an entire population of neurons by using two different quantification techniques and compared with the population firing rate. We find that the task timing was represented in the temporal structure of significant spike synchronization at the population level. During practice, the temporal structure of synchrony was shaped, with synchrony becoming stronger and more localized in time during late experimental sessions, in parallel with an improvement in behavioral performance. Concurrently, the average population firing rate mainly decreased. Performance optimization through practice might therefore be achieved by boosting the computational contribution of spike synchrony, allowing an overall reduction in population activity. PMID- 19812341 TI - Cocaine exposure in utero alters synaptic plasticity in the medial prefrontal cortex of postnatal rats. AB - Cocaine exposure during pregnancy causes abnormality in fetal brain development, leading to cognitive dysfunction of the offspring, but the underlying cellular mechanism remains mostly unclear. In this study, we examined synaptic functions in the medial prefrontal cortex (mPFC) of postnatal rats that were exposed to cocaine in utero, using whole-cell recording from mPFC layer V pyramidal neurons in acute brain slices. Cocaine exposure in utero resulted in a facilitated activity-induced long-term potentiation (LTP) of excitatory synapses on these pyramidal neurons and an elevated neuronal excitability in postnatal rat pups after postnatal day 15 (P15). This facilitated LTP could be primarily attributed to the reduction of GABAergic inhibition. Biochemical assays of isolated mPFC tissue from postnatal rats further showed that cocaine exposure in utero caused a marked reduction in the surface expression of GABA(A) receptor subunits alpha1, beta2, and beta3, but had no effect on glutamate receptor subunit GluR1. Both facilitated LTP and reduced surface expression of GABA(A) receptors persisted in rats up to at least P42. Finally, the behavioral consequence of cocaine exposure in utero was reflected by the reduction in the sensitivity of locomotor activity in postnatal rats to cocaine and the dopamine receptor agonist apomorphine. Since the mPFC is an important part of the reward circuit in the rat brain and plays important roles in cognitive functions, these findings offer new insights into the cellular mechanism underlying the adverse effects of cocaine exposure in utero on brain development and cognitive functions. PMID- 19812342 TI - Intracranial EEG reveals a time- and frequency-specific role for the right inferior frontal gyrus and primary motor cortex in stopping initiated responses. AB - Inappropriate response tendencies may be stopped via a specific fronto/basal ganglia/primary motor cortical network. We sought to characterize the functional role of two regions in this putative stopping network, the right inferior frontal gyrus (IFG) and the primary motor cortex (M1), using electocorticography from subdural electrodes in four patients while they performed a stop-signal task. On each trial, a motor response was initiated, and on a minority of trials a stop signal instructed the patient to try to stop the response. For each patient, there was a greater right IFG response in the beta frequency band ( approximately 16 Hz) for successful versus unsuccessful stop trials. This finding adds to evidence for a functional network for stopping because changes in beta frequency activity have also been observed in the basal ganglia in association with behavioral stopping. In addition, the right IFG response occurred 100-250 ms after the stop signal, a time range consistent with a putative inhibitory control process rather than with stop-signal processing or feedback regarding success. A downstream target of inhibitory control is M1. In each patient, there was alpha/beta band desynchronization in M1 for stop trials. However, the degree of desynchronization in M1 was less for successfully than unsuccessfully stopped trials. This reduced desynchronization on successful stop trials could relate to increased GABA inhibition in M1. Together with other findings, the results suggest that behavioral stopping is implemented via synchronized activity in the beta frequency band in a right IFG/basal ganglia network, with downstream effects on M1. PMID- 19812344 TI - Involvement of the thalamocortical loop in the spontaneous switching of percepts in auditory streaming. AB - Perceptual grouping of successive frequency components, namely, auditory streaming, is essential for auditory scene analysis. Prolonged listening to an unchanging triplet-tone sequence produces a series of illusory switches between a single coherent stream (S1) and two distinct streams (S2). The predominant percept depends on the frequency difference (Deltaf) between high and low tones. Here, we combined the use of different Deltafs with an event-related fMRI design to identify whether the temporal dynamics of brain activity differs depending on the direction of perceptual switches. The results demonstrated that the activity of the medial geniculate body (MGB) in the thalamus occurred earlier during switching from nonpredominant to predominant percepts, whereas that of the auditory cortex (AC) occurred earlier during switching from predominant to nonpredominant percepts, regardless of Deltaf. The asymmetry of temporal precedence indicates that the MGB and AC activations play different roles in perceptual switching and depend on perceptual predominance rather than on S1 and S2 percepts per se. Our results suggest that feedforward and feedback processes in the thalamocortical loop are involved in the formation of percepts in auditory streaming. PMID- 19812343 TI - Disruption of functional connectivity in clinically normal older adults harboring amyloid burden. AB - Amyloid deposition is present in 20-50% of nondemented older adults yet the functional consequences remain unclear. The current study found that amyloid accumulation is correlated with functional disruption of the default network as measured by intrinsic activity correlations. Clinically normal participants (n = 38, aged 60-88 years) were characterized using (11)C-labeled Pittsburgh Compound B positron emission tomography imaging to estimate fibrillar amyloid burden and, separately, underwent functional magnetic resonance imaging (fMRI). The integrity of the default network was estimated by correlating rest-state fMRI time courses extracted from a priori regions including the posterior cingulate, lateral parietal, and medial prefrontal cortices. Clinically normal participants with high amyloid burden displayed significantly reduced functional correlations within the default network relative to participants with low amyloid burden. These reductions were also observed when amyloid burden was treated as a continuous, rather than a dichotomous, measure and when controlling for age and structural atrophy. Whole-brain analyses initiated by seeding the posterior cingulate cortex, a region of high amyloid burden in Alzheimer's disease, revealed significant disruption in the default network including functional disconnection of the hippocampal formation. PMID- 19812345 TI - Autocrine activation of neuronal NMDA receptors by aspartate mediates dopamine- and cAMP-induced CREB-dependent gene transcription. AB - cAMP can stimulate the transcription of many activity-dependent genes via activation of the transcription factor, cAMP response element-binding protein (CREB). However, in mouse cortical neuron cultures, prior to synaptogenesis, neither cAMP nor dopamine, which acts via cAMP, stimulated CREB-dependent gene transcription when NR2B-containing NMDA receptors (NMDARs) were blocked. Stimulation of transcription by cAMP was potentiated by inhibitors of excitatory amino acid uptake, suggesting a role for extracellular glutamate or aspartate in cAMP-induced transcription. Aspartate was identified as the extracellular messenger: enzymatic scavenging of l-aspartate, but not glutamate, blocked stimulation of CREB-dependent gene transcription by cAMP; moreover, cAMP induced aspartate but not glutamate release. Together, these results suggest that cAMP acts via an autocrine or paracrine pathway to release aspartate, which activates NR2B-containing NMDARs, leading to Ca(2+) entry and activation of transcription. This cAMP/aspartate/NMDAR signaling pathway may mediate the effects of transmitters such as dopamine on axon growth and synaptogenesis in developing neurons or on synaptic plasticity in mature neural networks. PMID- 19812346 TI - HSE management standards and stress-related work outcomes. AB - BACKGROUND: The UK Health and Safety Executive's (HSE) Management Standards (MS) approach has been developed to help organizations manage potential sources of work-related stress. Although there is general support for the assessment model adopted by this approach, to date, there has been no empirical investigation of the relationship between the actual MS (as measured by the final revised version of the HSE Indicator Tool) and stress-related work outcomes. AIMS: To investigate the relationship between the HSE MS and the following stress-related work outcomes: 'job satisfaction', job-related anxiety and depression and errors/near misses. METHODS: An anonymous cross-sectional questionnaire was distributed by either e-mail or post to all employees within a community-based Health and Social Services Trust. Respondents completed the HSE Indicator Tool, a job-related anxiety and depression scale, a job satisfaction scale and an aggregated measure of the number of errors/near misses witnessed. Associations between the HSE Indicator Tool responses and stress-related work outcomes were analysed with regression statistics. RESULTS: A total of 707 employees completed the questionnaire, representing a low response rate of 29%. Controlling for age, gender and contract type, the HSE MS (as measured by the HSE Indicator Tool) were positively associated with job satisfaction and negatively associated with 'job related anxiety', 'job-related depression' and 'witnessed errors/near misses'. CONCLUSIONS: This study provides empirical evidence to support the use of the MS approach in tackling workplace stress. PMID- 19812347 TI - Musculoskeletal symptoms in pharmaceutical sales representatives. AB - BACKGROUND: Musculoskeletal disorders (MSDs) are a leading cause of work-related ill health. Existing literature indicates that pharmaceutical sales representatives (PSRs) report a high prevalence of MSDs, possibly exacerbated by the nature of work (prolonged driving and manual handling). In addition, they experience difficulty in accessing occupational health services. AIMS: To assess the prevalence of musculoskeletal symptoms and associated risk factors among PSRs in order to assist their occupational health management through raising risk awareness. METHODS: A self-completed questionnaire distributed to 205 PSRs within a UK pharmaceutical company was used to assess the prevalence of musculoskeletal symptoms, psychosocial factors, work tasks undertaken and company car use. To assist understanding of work tasks and organizational factors, semi-structured interviews were undertaken with a sample of 12 key personnel. RESULTS: The questionnaire response rate was 68%. PSRs reported high mileage and 100% reported working from the car in a typical day. Forty-seven per cent reported both manual handling for > or = 4 h/day and 'often' or 'sometimes' working from the car. Fifty-seven per cent reported low back symptoms in the last 12 months. Interview data revealed issues relating to car choice, storage in the boot and working from the car, which should be considered when developing priorities for preventive management of MSDs. CONCLUSIONS: Musculoskeletal symptoms appear to be a problem for PSRs, with risk factors reported as prolonged driving, sitting in the car, working from the car and manual handling. Interventions to facilitate their occupational health management should focus on raising awareness of the risks of prolonged driving and working from the car. PMID- 19812348 TI - Identification of the human cytochrome P450 enzymes involved in the two oxidative steps in the bioactivation of clopidogrel to its pharmacologically active metabolite. AB - The aim of the current study is to identify the human cytochrome P450 (P450) isoforms involved in the two oxidative steps in the bioactivation of clopidogrel to its pharmacologically active metabolite. In the in vitro experiments using cDNA-expressed human P450 isoforms, clopidogrel was metabolized to 2-oxo clopidogrel, the immediate precursor of its pharmacologically active metabolite. CYP1A2, CYP2B6, and CYP2C19 catalyzed this reaction. In the same system using 2 oxo-clopidogrel as the substrate, detection of the active metabolite of clopidogrel required the addition of glutathione to the system. CYP2B6, CYP2C9, CYP2C19, and CYP3A4 contributed to the production of the active metabolite. Secondly, the contribution of each P450 involved in both oxidative steps was estimated by using enzyme kinetic parameters. The contribution of CYP1A2, CYP2B6, and CYP2C19 to the formation of 2-oxo-clopidogrel was 35.8, 19.4, and 44.9%, respectively. The contribution of CYP2B6, CYP2C9, CYP2C19, and CYP3A4 to the formation of the active metabolite was 32.9, 6.76, 20.6, and 39.8%, respectively. In the inhibition studies with antibodies and selective chemical inhibitors to P450s, the outcomes obtained by inhibition studies were consistent with the results of P450 contributions in each oxidative step. These studies showed that CYP2C19 contributed substantially to both oxidative steps required in the formation of clopidogrel active metabolite and that CYP3A4 contributed substantially to the second oxidative step. These results help explain the role of genetic polymorphism of CYP2C19 and also the effect of potent CYP3A inhibitors on the pharmacokinetics and pharmacodynamics of clopidogrel in humans and on clinical outcomes. PMID- 19812349 TI - Gene regulation of CYP4F11 in human keratinocyte HaCaT cells. AB - Mechanisms regulating CYP4F genes remain under investigation, although characterization of CYP4F regulatory modalities would facilitate the discovery of new drug targets. This present study shows that all-trans- and 9-cis-retinoic acids can inhibit CYP4F11 expression in human keratinocyte-derived HaCaT cells. Transrepression of many genes by retinoic acids is mediated by interactions between retinoid receptors and the activator protein 1 (AP-1) complex. Proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha) and interleukin 1beta, which can activate the AP-1 complex, induce CYP4F11 transcription in HaCaT cells. The c-Jun N-terminal kinase (JNK)-specific inhibitor 1,9-pyrazoloanthrone (SP600125) blocked the induction of CYP4F11 by both cytokines, indicating involvement of the JNK pathway. Furthermore, TNF-alpha failed to induce CYP4F11 transcription when HaCaT cells were preincubated with retinoic acids. Retinoic acids are ligands for the retinoic acid receptors (RARs) and the retinoid X receptors (RXRs). The RXR agonist 6-(1(3,5,5,8,8-pentamethyl-5,6,7,8 tetrahydronaphthalen-2-yl)cyclopropyl) nicotinic acid (LG268) greatly induced CYP4F11 transcription, whereas the RAR agonist 4-(2-(5,6,7,8-tetrahydro-5,5,8,8 tetramethyl-2-naphthalenyl)-1-propenyl)benzoic acid (TTNPB) markedly inhibited CYP4F11 transcription, indicating that down-regulation of CYP4F11 transcription by retinoic acid is mediated by RARs and may also be related to ligand competition for RXRs. Thus, the CYP4F11 gene is positively regulated by multiple signaling pathways in HaCaT keratinocytes, including RXR and JNK signaling pathways. PMID- 19812350 TI - In vitro metabolism of haloperidol and sila-haloperidol: new metabolic pathways resulting from carbon/silicon exchange. AB - The neurotoxic side effects observed for the neuroleptic agent haloperidol have been associated with its pyridinium metabolite. In a previous study, a silicon analog of haloperidol (sila-haloperidol) was synthesized, which contains a silicon atom instead of the carbon atom in the 4-position of the piperidine ring. In the present study, the phase I metabolism of sila-haloperidol and haloperidol was studied in rat and human liver microsomes. The phase II metabolism was studied in rat, dog, and human hepatocytes and also in liver microsomes supplemented with UDP-glucuronic acid (UDPGA). A major metabolite of haloperidol, the pyridinium metabolite, was not formed in the microsomal incubations with sila haloperidol. For sila-haloperidol, three metabolites originating from opening of the piperidine ring were observed, a mechanism that has not been observed for haloperidol. One of the significant phase II metabolites of haloperidol was the glucuronide of the hydroxy group bound to the piperidine ring. For sila haloperidol, the analogous metabolite was not observed in the hepatocytes or in the liver microsomal incubations containing UDPGA. If silanol (SiOH) groups are not glucuronidated, introducing silanol groups in drug molecules could provide an opportunity to enhance the hydrophilicity without allowing for direct phase II metabolism. To provide further support for the observed differences in metabolic pathways between haloperidol and sila-haloperidol, the metabolism of another pair of C/Si analogs was studied, namely, trifluperidol and sila-trifluperidol. These studies showed the same differences in metabolic pathways as between sila haloperidol and haloperidol. PMID- 19812351 TI - Tamoxifen-induced adduct formation and cell stress in human endometrial glands. AB - The beneficial effects of tamoxifen in the prevention and treatment of breast cancer are compromised by an increased risk of endometrial polyps, hyperplasia, and cancer. Tamoxifen is metabolized to an array of metabolites with estrogenic effects but also to reactive intermediates that may form protein and DNA adducts. The aim of this study was to investigate cellular [(3)H]tamoxifen adduct formation by light microscopic autoradiography and cell stress by immunohistochemical analysis of glucose-regulating protein 78 (GRP78), nuclear factor kappaB (NF-kappaB), and caspase 3 in human endometrial explants after short-term incubation with tamoxifen. The cellular expression of tamoxifen metabolizing enzymes in human endometrial biopsy samples was also determined by immunohistochemistry. The results showed selective [(3)H]tamoxifen adduct formation in glandular and surface epithelia after incubation with a nontoxic concentration of [(3)H]tamoxifen (6 nM). There was also a selective expression of the endoplasmic reticulum stress chaperone GRP78 and activated caspase 3 at these sites after incubation with cytotoxic concentrations of tamoxifen (10-100 microM). The cell stress was preferentially observed in samples from women in the proliferative menstrual phase. No treatment-related expression of NF-kappaB was observed. Constitutive expression of the tamoxifen-metabolizing enzymes CYP1B1, CYP2A6, CYP2B6, CYP2C8/9/19, CYP2D6, and SULT2A1 in glandular and surface epithelia was shown, but there was a large interindividual variation. The colocalization of [(3)H]tamoxifen adducts, expression of GRP78, caspase 3, and tamoxifen-metabolizing enzymes in human glandular and surface epithelia suggest a local bioactivation of tamoxifen at these sites and that epithelial cells are early target sites for tamoxifen-induced cell stress. PMID- 19812352 TI - Identification of multiple glutathione conjugates of 8-amino- 2-methyl-4-phenyl 1,2,3,4-tetrahydroisoquinoline maleate (nomifensine) in liver microsomes and hepatocyte preparations: evidence of the bioactivation of nomifensine. AB - 8-Amino-2-methyl-4-phenyl-1,2,3,4-tetrahydroisoquinoline maleate (nomifensine), an antidepressant drug, was withdrawn from the market because of increased incidence of hemolytic anemia, as well as kidney and liver toxicity. Although the nature of the potentially reactive metabolites formed after nomifensine metabolism remains unknown and no glutathione (GSH) adducts of these nomifensine reactive metabolites have been reported, bioactivation has been postulated as a potential mechanism for the toxicity of nomifensine. This study was conducted to probe the potential bioactivation pathways of nomifensine in human and animal hepatocytes and in liver microsomes using GSH as a trapping agent. Two types of GSH conjugates were characterized by liquid chromatography/tandem mass spectrometry: 1) aniline oxidation followed by GSH conjugation leading to the formation of nomifensine-GSH sulfinamides (M1 and M2); and 2) arene oxidation followed by GSH conjugation yielding a range of arene C-linked GSH adducts (M3 M9). Nine GSH adducts (M1-M9) were identified in liver microsomes of humans, dogs, monkeys, and rats and in human and rat hepatocytes. In dog hepatocyte preparations, six GSH adducts (M1-M6) were identified. The GSH adducts in dog and rat liver microsomes were formed primarily through aniline and arene oxidation, respectively. Both pathways contributed significantly to the formation of the GSH adducts in human and monkey liver microsomes. The bioactivation pathways proposed here account for the formation of the observed GSH conjugates. These investigations have confirmed the aniline and the arene groups in nomifensine as potential toxicophores capable of generating reactive intermediates. PMID- 19812353 TI - Protease-activated receptor 2-mediated protection of myocardial ischemia reperfusion injury: role of transient receptor potential vanilloid receptors. AB - Activation of the protease-activated receptor 2 (PAR2) or the transient receptor potential vanilloid type 1 (TRPV1) channels expressed in cardiac sensory afferents containing calcitonin gene-related peptide (CGRP) and/or substance P (SP) has been proposed to play a protective role in myocardial ischemia reperfusion (I/R) injury. However, the interaction between PAR2 and TRPV1 is largely unknown. Using gene-targeted TRPV1-null mutant (TRPV1(-/-)) or wild-type (WT) mice, we test the hypothesis that TRPV1 contributes to PAR2-mediated cardiac protection via increasing the release of CGRP and SP. Immunofluorescence labeling showed that TRPV1 coexpressed with PAR2, PKC-epsilon, or PKAc in cardiomyocytes, cardiac blood vessels, and perivascular nerves in WT but not TRPV1(-/-) hearts. WT or TRPV1(-/-) hearts were Langendorff perfused with the selective PAR2 agonist, SLIGRL, in the presence or absence of various antagonists, followed by 35 min of global ischemia and 40 min of reperfusion (I/R). The recovery rate of coronary flow, the maximum rate of left ventricular pressure development, left ventricular end-diastolic pressure, and left ventricular developed pressure were evaluated after I/R. SLIGRL improved the recovery of hemodynamic parameters, decreased lactate dehydrogenase release, and reduced the infarct size in both WT and TRPV1(-/-) hearts (P < 0.05). The protection of SLIGRL was significantly surpassed for WT compared with TRPV1(-/-) hearts (P < 0.05). CGRP(8-37), a selective CGRP receptor antagonist, RP67580, a selective neurokinin-1 receptor antagonist, PKC-epsilon V1-2, a selective PKC-epsilon inhibitor, or H-89, a selective PKA inhibitor, abolished SLIGRL protection by inhibiting the recovery of the rate of coronary flow, maximum rate of left ventricular pressure development, and left ventricular developed pressure, and increasing left ventricular end-diastolic pressure in WT but not TRPV1(-/-) hearts. Radioimmunoassay showed that SLIGRL increased the release of CGRP and SP in WT but not TRPV1(-/-) hearts (P < 0.05), which were prevented by PKC-epsilon V1-2 and H-89. Thus our data show that PAR2 activation improves cardiac recovery after I/R injury in WT and TRPV1(-/-) hearts, with a greater effect in the former, suggesting that PAR2-mediated protection is TRPV1 dependent and independent, and that dysfunctional TRPV1 impairs PAR2 action. PAR2 activation of the PKC-epsilon or PKA pathway stimulates or sensitizes TRPV1 in WT hearts, leading to the release of CGRP and SP that contribute, at least in part, to PAR2-induced cardiac protection against I/R injury. PMID- 19812354 TI - Absorption kinetics are a key factor regulating postprandial protein metabolism in response to qualitative and quantitative variations in protein intake. AB - We have previously demonstrated that increasing the habitual protein intake widened the gap in nutritional quality between proteins through mechanisms that are not yet fully understood. We hypothesized that the differences in gastrointestinal kinetics between dietary proteins were an important factor affecting their differential response to an increased protein intake. To test this hypothesis, we built a 13-compartment model providing integrative insight into the sequential dynamics of meal nitrogen (Nm) absorption, splanchnic uptake, and metabolism, and subsequent peripheral transfer and deposition. The model was developed from data on postprandial Nm kinetics in certain accessible pools, obtained from subjects having ingested a (15)N-labeled milk or soy protein meal, after adaptation to normal (NP) or high (HP) protein diets. The faster absorption of Nm after soy vs. milk caused its earlier and stronger splanchnic delivery, which favored its local catabolic utilization (up to +30%) and limited its peripheral accretion (down to -20%). Nm absorption was also accelerated after HP vs. NP adaptation, and this kinetic effect accounted for most of the HP-induced increase (up to +20%) in splanchnic Nm catabolic use, and the decrease (down to 25%) in peripheral Nm anabolic utilization. The HP-induced acceleration in Nm absorption was more pronounced with soy than with milk, as were the HP effects on Nm regional metabolism. Our integrative approach identified Nm absorption kinetics, which exert a direct and lasting impact on Nm splanchnic catabolic use and peripheral delivery, as being critical in adaptation to both qualitative and quantitative changes in protein intake. PMID- 19812355 TI - The role of aldosterone in mediating the dependence of angiotensin hypertension on IL-6. AB - Knockout (KO) of IL-6 has been shown to attenuate ANG II hypertension, and mineralocorticoid receptors (MR) have been reported to contribute to the increase in IL-6 during acute ANG II infusion. This study determined whether that MR action is sustained with chronic ANG II infusion and whether it plays a role in mediating ANG II hypertension. ANG II infusion (90 ng/min) increased plasma IL-6 from 1.6 +/- 0.6 to 22.7 +/- 2.2 and 19.9 +/- 3.2 pg/ml on days 7 and 14, respectively, and chronic MR blockade with spironolactone attenuated that only at day 7 (7.2 +/- 2.2 pg/ml). ANG II increased MAP (19 h/day with telemetry) approximately 40 mmHg, but in ANG II+spironolactone mice (25 or 50 mg*kg(-1)*day( 1)), mean arterial pressure (MAP) was not significantly different despite a tendency for lower pressure the first 6 days. To isolate further the mineralocorticoid link to IL-6 and blood pressure, DOCA-salt hypertension was induced in IL-6 KO and wild-type (WT) mice. Plasma IL-6 increased from 4.1 +/- 1.7 to 34.5 +/- 7.0 pg/ml by day 7 of DOCA treatment in the WT mice but was back to control levels by day 14. An IL-6 bioassay using the murine B9, B-cell hybridoma cell line demonstrated that plasma IL-6 measurements reflected actual IL-6 bioactivity. The hypertension was not different and virtually superimposable in WT vs. IL-6 KO mice, averaging 145 +/- 2 and 144 +/- 3 mmHg, respectively. Both experiments confirm chronic stimulation of IL-6 by mineralocorticoids but show that it is transient. In addition, IL-6 was not required for mineralocorticoid hypertension. This suggests that aldosterone contributes to the increase in plasma IL-6 in the early stage of ANG II hypertension but that the blood pressure actions of IL-6 in that model are linked most likely to ANG II rather than aldosterone. PMID- 19812356 TI - Ca2+ cycling in cardiomyocytes from a high-performance reptile, the varanid lizard (Varanus exanthematicus). AB - The varanid lizard possesses one of the largest aerobic capacities among reptiles with maximum rates of oxygen consumption that are twice that of other lizards of comparable sizes at the same temperature. To support this aerobic capacity, the varanid heart possesses morphological adaptations that allow the generation of high heart rates and blood pressures. Specializations in excitation-contraction coupling may also contribute to the varanids superior cardiovascular performance. Therefore, we investigated the electrophysiological properties of the l-type Ca(2+) channel and the Na(+)/Ca(2+) exchanger (NCX) and the contribution of the sarcoplasmic reticulum to the intracellular Ca(2+) transient (Delta[Ca(2+)](i)) in varanid lizard ventricular myocytes. Additionally, we used confocal microscopy to visualize myocytes and make morphological measurements. Lizard ventricular myocytes were found to be spindle-shaped, lack T-tubules, and were approximately 190 microm in length and 5-7 microm in width and depth. Cardiomyocytes had a small cell volume ( approximately 2 pL), leading to a large surface area-to volume ratio (18.5), typical of ectothermic vertebrates. The voltage sensitivity of the l-type Ca(2+) channel current (I(Ca)), steady-state activation and inactivation curves, and the time taken for recovery from inactivation were also similar to those measured in other reptiles and teleosts. However, transsarcolemmal Ca(2+) influx via reverse mode Na(+)/Ca(2+) exchange current was fourfold higher than most other ectotherms. Moreover, pharmacological inhibition of the sarcoplasmic reticulum led to a 40% reduction in the Delta[Ca(2+)](i) amplitude, and slowed the time course of decay. In aggregate, our results suggest varanids have an enhanced capacity to transport Ca(2+) through the Na(+)/Ca(2+) exchanger, and sarcoplasmic reticulum suggesting specializations in excitation contraction coupling may provide a means to support high cardiovascular performance. PMID- 19812357 TI - Plasma hyperosmolality elevates the internal temperature threshold for active thermoregulatory vasodilation during heat stress in humans. AB - Plasma hyperosmolality delays the response in skin blood flow to heat stress by elevating the internal temperature threshold for cutaneous vasodilation. This elevation could be because of a delayed onset of cutaneous active vasodilation and/or to persistent cutaneous active vasoconstriction. Seven healthy men were infused with either hypertonic (3% NaCl) or isotonic (0.9% NaCl) saline and passively heated by immersing their lower legs in 42 degrees C water for 60 min (room temperature, 28 degrees C; relative humidity, 40%). Skin blood flow was monitored via laser-Doppler flowmetry at sites pretreated with bretylium tosylate (BT) to block sympathetic vasoconstriction selectively and at adjacent control sites. Plasma osmolality was increased by approximately 13 mosmol/kgH(2)O following hypertonic saline infusion and was unchanged following isotonic saline infusion. The esophageal temperature (T(es)) threshold for cutaneous vasodilation at untreated sites was significantly elevated in the hyperosmotic state (37.73 +/ 0.11 degrees C) relative to the isosmotic state (36.63 +/- 0.12 degrees C, P < 0.001). A similar elevation of the T(es) threshold for cutaneous vasodilation was observed between osmotic conditions at the BT-treated sites (37.74 +/- 0.18 vs. 36.67 +/- 0.07 degrees C, P < 0.001) as well as sweating. These results suggest that the hyperosmotically induced elevation of the internal temperature threshold for cutaneous vasodilation is due primarily to an elevation in the internal temperature threshold for the onset of active vasodilation, and not to an enhancement of vasoconstrictor activity. PMID- 19812358 TI - Identification of renal transporters involved in sulfate excretion in marine teleost fish. AB - Sulfate (SO(4)(2-)) is the second most abundant anion in seawater (SW), and excretion of excess SO(4)(2-) from ingested SW is essential for marine fish to survive. Marine teleosts excrete SO(4)(2-) via the urine produced in the kidney. The SO(4)(2-) transporter that secretes and concentrates SO(4)(2-) in the urine has not previously been identified. Here, we have identified and characterized candidates for the long-sought transporters. Using sequences from the fugu database, we have cloned cDNA fragments of all transporters belonging to the Slc13 and Slc26 families from mefugu (Takifugu obscurus). We compared Slc13 and Slc26 mRNA expression in the kidney between freshwater (FW) and SW mefugu. Among 14 clones examined, the expression of a Slc26a6 paralog (mfSlc26a6A) was the most upregulated (30-fold) in the kidney of SW mefugu. Electrophysiological analyses of Xenopus oocytes expressing mfSlc26a6A, mfSlc26a6B, and mouse Slc26a6 (mSlc26a6) demonstrated that all transporters mediate electrogenic Cl(-)/SO(4)(2 ), Cl(-)/oxalate(2-), and Cl(-)/nHCO(3)(-) exchanges and electroneutral Cl( )/formate(-) exchange. Two-electrode voltage-clamp experiments demonstrated that the SO(4)(2-)-elicited currents of mfSlc26a6A is quite large (approximately 35 microA at +60 mV) and 50- to 200-fold higher than those of mfSlc26a6B and mSlc26a6. Conversely, the currents elicited by oxalate and HCO(3)(-) are almost identical among mfSlc26a6A, mfSlc26a6B, and mSlc26a6. Kinetic analysis revealed that mfSlc26a6A has the highest SO(4)(2-) affinity as well as capacity. Immunohistochemical analyses demonstrated that mfSlc26a6A localizes to the apical (brush-border) region of the proximal tubules. Together, these findings suggest that mfSlc26a6A is the most likely candidate for the major apical SO(4)(2-) transporter that mediates SO(4)(2-) secretion in the kidney of marine teleosts. PMID- 19812359 TI - CaMKK is an upstream signal of AMP-activated protein kinase in regulation of substrate metabolism in contracting skeletal muscle. AB - Multiple signals have been shown to be involved in regulation of fatty acid (FA) and glucose metabolism in contracting skeletal muscle. This study aimed to determine whether a Ca(2+)-stimulated kinase, CaMKK, is involved in regulation of contraction-induced substrate metabolism and whether it does so in an AMP activated protein kinase (AMPK)-dependent manner. Rat hindlimbs were perfused at rest (n = 16), with 3 mM caffeine (n = 15), with 2 mM 5-aminoimidazole-4 carboxamide 1-beta-d-ribofuranoside (AICAR; n = 16), or during moderate-intensity muscle contraction (MC; n = 14) and with or without 5 microM STO-609, a CaMKK inhibitor. FA uptake and oxidation increased (P < 0.05) 64% and 71% by caffeine, 42% and 93% by AICAR, and 65% and 143% by MC. STO-609 abolished (P < 0.05) caffeine- and MC-induced FA uptake and oxidation but had no effect with AICAR treatment. Glucose uptake increased (P < 0.05) 104% by caffeine, 85% by AICAR, and 130% by MC, and STO-609 prevented the increase in glucose uptake in caffeine and muscle contraction groups. CaMKKbeta activity increased (P < 0.05) 113% by caffeine treatment and 145% by MC but was not affected by AICAR treatment. STO 609 prevented the caffeine- and MC-induced increase in CaMKKbeta activity. Caffeine, AICAR, and MC increased (P < 0.05) AMPKalpha2 activity by 295%, 11 fold, and 7-fold but did not affect AMPKalpha1 activity. STO-609 decreased (P < 0.05) AMPKalpha2 activity induced by caffeine treatment and MC by 60% and 61% but did not affect AICAR-induced activity. Plasma membrane transport protein content of CD36 and glucose transporter 4 (GLUT4) increased (P < 0.05) with caffeine, AICAR, and MC, and STO-609 prevented caffeine- and MC-induced increases in protein content. These results show the importance of Ca(2+)-dependent signaling via CaMKK activation in the regulation of substrate uptake and FA oxidation in contracting rat skeletal muscle and agree with the notion that CaMKK is an upstream kinase of AMPK in the regulation of substrate metabolism in skeletal muscle. PMID- 19812360 TI - Estrogen replacement restores flow-induced vasodilation in coronary arterioles of aged and ovariectomized rats. AB - The risk for cardiovascular disease (CVD) increases with advancing age; however, the age at which CVD risk increases significantly is delayed by more than a decade in women compared with men. This cardioprotection, which women experience until menopause, is presumably due to the presence of ovarian hormones, in particular, estrogen. The purpose of this study was to determine how age and ovarian hormones affect flow-induced vasodilation in the coronary resistance vasculature. Coronary arterioles were isolated from young (6 mo), middle-aged (14 mo), and old (24 mo) intact, ovariectomized (OVX), and ovariectomized + estrogen replaced (OVE) female Fischer-344 rats to assess flow-induced vasodilation. Advancing age impaired flow-induced dilation of coronary arterioles (young: 50 +/ 4 vs. old: 34 +/- 6; % relaxation). Ovariectomy reduced flow-induced dilation in arterioles from young females, and estrogen replacement restored vasodilation to flow. In aged females, flow-induced vasodilation of arterioles was unaltered by OVX; however, estrogen replacement improved flow-induced dilation by approximately 160%. The contribution of nitric oxide (NO) to flow-induced dilation, assessed by nitric oxide synthase (NOS) inhibition with N(G)-nitro-l arginine methyl ester (l-NAME), declined with age. l-NAME did not alter flow induced vasodilation in arterioles from OVX rats, regardless of age. In contrast, l-NAME reduced flow-induced vasodilation of arterioles from estrogen-replaced rats at all ages. These findings indicate that the age-induced decline of flow induced, NO-mediated dilation in coronary arterioles of female rats is related, in part, to a loss of ovarian estrogen, and estrogen supplementation can improve flow-induced dilation, even at an advanced age. PMID- 19812361 TI - In utero exposure to benzene disrupts fetal hematopoietic progenitor cell growth via reactive oxygen species. AB - It is hypothesized that the increasing incidence of childhood leukemia may be due to in utero exposure to environmental pollutants, such as benzene, but the mechanisms involved remain unknown. We hypothesize that reactive oxygen species (ROS) contribute to the deregulation of fetal hematopoiesis caused by in utero benzene exposure. To evaluate this hypothesis, pregnant C57Bl/6N mice were exposed to benzene or polyethylene glycol-conjugated catalase (PEG-catalase) (antioxidative enzyme) and benzene. Colony formation assays on fetal liver cells were performed to measure erythroid and myeloid progenitor cell growth potential. The presence of ROS in CD117(+) fetal liver cells was measured by flow cytometric analysis. Oxidative cellular damage was assessed by Western blot analysis of 4 hydroxynonenol (4-HNE) and nitrotyrosine products, as well as reduced to oxidized glutathione ratios. Alterations in the redox-sensitive signaling pathway nuclear factor-kappa-light-chain-enhancer of activated B cells (NF-kappaB) were measured by Western blot analysis of Inhibitor of NF-kB-alpha (IkappaB-alpha) protein levels in fetal liver tissue. In utero exposure to benzene caused a significant increase in ROS production and significantly altered fetal liver erythroid and myeloid colony numbers but did not increase the levels of 4-HNE or nitrotyrosine products or alter reduced to oxidized glutathione ratios. However, in utero exposure to benzene did cause a significant decrease in fetal liver IkappaB-alpha protein levels, suggesting activation of the NF-kappaB pathway. Benzene-induced ROS formation, abnormal colony growth, and decreased IkappaB-alpha levels were all abrogated by pretreatment with PEG-catalase. These results suggest that ROS play a key role in the development of in utero-initiated benzene toxicity potentially through disruption of hematopoietic cell signaling pathways. PMID- 19812362 TI - Age-dependent susceptibility to manganese-induced neurological dysfunction. AB - Chronic exposure to manganese (Mn) produces a spectrum of cognitive and behavioral deficits associated with a neurodegenerative disorder resembling Parkinson's disease. The effects of high-dose exposure to Mn in occupational cohorts and in adult rodent models of the disease are well described but much less is known about the behavioral and neurochemical effects of Mn in the developing brain. We therefore exposed C57Bl/6 mice to Mn by intragastric gavage as juveniles, adults, or both, postulating that mice exposed as juveniles and then again as adults would exhibit greater neurological and neurochemical dysfunction than mice not preexposed as juveniles. Age- and sex-dependent vulnerability to changes in locomotor function was detected, with juvenile male mice displaying the greatest sensitivity, characterized by a selective increase in novelty-seeking and hyperactive behaviors. Adult male mice preexposed as juveniles had a decrease in total movement and novelty-seeking behavior, and no behavioral changes were detected in female mice. Striatal dopamine levels were increased in juvenile mice but were decreased in adult preexposed as juveniles. Levels of Mn, Fe, and Cu were determined by inductively coupled plasma-mass spectrometry, with the greatest accumulation of Mn detected in juvenile mice in the striatum, substantia nigra (SN), and cortex. Only modest changes in Fe and Cu were detected in Mn-treated mice, primarily in the SN. These results reveal that developing mice are more sensitive to Mn than adult animals and that Mn exposure during development enhances behavioral and neurochemical dysfunction relative to adult animals without juvenile exposure. PMID- 19812363 TI - Reduction of glutamatergic neurotransmission by prolonged exposure to dieldrin involves NMDA receptor internalization and metabotropic glutamate receptor 5 downregulation. AB - Dieldrin was previously used as a pesticide. Although its use has been discontinued, humans are still exposed to it due to its high environmental persistence and because it accumulates in the adipose tissue of animals. Acute exposure to dieldrin provokes convulsions due to its antagonism on the gamma aminobutyric acid-A (GABA(A)) receptor. However, little is known about the effects of low chronic exposure to this pollutant. In the present work, we use primary cultures of cortical neurons to study the mechanisms involved in the toxic action of dieldrin. We found that 2 and 6 days in vitro (DIV) exposure to a subcytotoxic concentration (60nM) of dieldrin reduced the increase in intracellular calcium concentration ([Ca(2+)](i)) and the excitotoxicity caused by glutamate. Exposure to dieldrin for 6 DIV induced N-methyl-D-aspartate receptor (NMDAR) internalization and reduced metabotropic glutamate receptor 5 (mGLUR5) levels. Double immunostaining for NMDAR and mGLUR5 showed that these receptors lose colocalization on the cell membrane in neurons treated with dieldrin. No changes were observed in receptor functionalities or receptor levels after 2 DIV of exposure to dieldrin. However, the increase in [Ca(2+)](i) induced by coactivation of NMDAR and mGLUR5 was significantly reduced. Thus, a functional interaction between the two receptors seems to play an important role in glutamate-induced excitotoxicity. We confirm that permanent blockade of the GABA(A) receptor by this persistent pesticide triggers adaptive neuronal changes consisting of a reduction of glutamatergic neurotransmission. This might explain the cognitive and learning deficits observed in animals after chronic treatment with dieldrin. PMID- 19812365 TI - Developmental exposure to manganese increases adult susceptibility to inflammatory activation of glia and neuronal protein nitration. AB - Chronic exposure to manganese (Mn) produces a neurodegenerative disorder affecting the basal ganglia characterized by reactive gliosis and expression of neuroinflammatory genes including inducible nitric oxide synthase (NOS2). Induction of NOS2 in glial cells causes overproduction of nitric oxide (NO) and injury to neurons that is associated with parkinsonian-like motor deficits. Inflammatory activation of glia is believed to be an early event in Mn neurotoxicity, but specific responses of microglia and astrocytes to Mn during development remain poorly understood. In this study, we investigated the effect of juvenile exposure to Mn on the activation of glia and production of NO in C57Bl/6J mice, postulating that developmental Mn exposure would lead to heightened sensitivity to gliosis and increased expression of NOS2 in adult mice exposed again later in life. Immunohistochemical analysis indicated that Mn exposure caused increased activation of both microglia and astrocytes in the striatum (St), globus pallidus (Gp), and substantia nigra pars reticulata (SNpr) of treated mice compared with controls. More robust activation of microglia was observed in juveniles, whereas astrogliosis was more prominent in adult mice preexposed during development. Co-immunofluorescence studies demonstrated increased expression of NOS2 in glia located in the Gp and SNpr. Additionally, greater increases in the level of 3-nitrotyrosine protein adducts were detected in dopamine- and cAMP-regulated phosphoprotein-32-positive neurons of the St of Mn-treated adult mice preexposed as juveniles. These data indicate that subchronic exposure to Mn during development leads to temporally distinct patterns of glial activation that result in elevated nitrosative stress in distinct populations of basal ganglia neurons. PMID- 19812364 TI - The role of hypoxia in 2-butoxyethanol-induced hemangiosarcoma. AB - To understand the molecular mechanisms underlying compound-induced hemangiosarcomas in mice, and therefore, their human relevance, a systems biology approach was undertaken using transcriptomics and Causal Network Modeling from mice treated with 2-butoxyethanol (2-BE). 2-BE is a hemolytic agent that induces hemangiosarcomas in mice. We hypothesized that the hemolysis induced by 2-BE would result in local tissue hypoxia, a well-documented trigger for endothelial cell proliferation leading to hemangiosarcoma. Gene expression data from bone marrow (BM), liver, and spleen of mice exposed to a single dose (4 h) or seven daily doses of 2-BE were used to develop a mechanistic model of hemangiosarcoma. The resulting mechanistic model confirms previous work proposing that 2-BE induces macrophage activation and inflammation in the liver. In addition, the model supports local tissue hypoxia in the liver and spleen, coupled with increased erythropoeitin signaling and erythropoiesis in the spleen and BM, and suppression of mechanisms that contribute to genomic stability, events that could be contributing factors to hemangiosarcoma formation. Finally, an immunohistochemistry method (Hypoxyprobe) demonstrated that tissue hypoxia was present in the spleen and BM. Together, the results of this study identify molecular mechanisms that initiate hemangiosarcoma, a key step in understanding safety concerns that can impact drug decision processes, and identified hypoxia as a possible contributing factor for 2-BE-induced hemangiosarcoma in mice. PMID- 19812366 TI - Cytotoxicity and regenerative proliferation as the mode of action for diuron induced urothelial carcinogenesis in the rat. AB - Diuron, a substituted urea herbicide, is carcinogenic to the urinary bladder of rats at high dietary levels. Its proposed carcinogenic mode of action (MOA) includes urothelial cytotoxicity and necrosis followed by regenerative cell proliferation and sustained urothelial hyperplasia. Cytotoxicity could be induced either by urinary solids or by chemical toxicity by diuron and/or metabolites excreted in the urine. Diuron was not genotoxic in a previous single-cell gel (comet) assay, but possible cross-linking activity remained to be evaluated. The present study explored the MOA of diuron and the effect of urinary acidification on the development of urothelial lesions. Male Wistar rats were fed diuron (2500 ppm, about 130 mg/kg of body weight) either with or without NH(4)Cl 10,000 ppm to acidify the urine. Reversibility of urothelial changes was also examined. The animals were euthanized after 15, 25, or 30 weeks. Diuron-fed rats had urinary amorphous precipitate and magnesium ammonium phosphate crystals similar to control animals. Groups treated with diuron + NH(4)Cl showed decreased urinary pH and reduced amounts of urinary crystals and precipitate. Urothelial necrosis and simple hyperplasia were observed by light microscopy and scanning electron microscopy both in diuron- and in diuron + NH(4)Cl-treated groups. Cytotoxicity and proliferative changes were mostly reversible. A modified comet assay developed in vitro with Chinese hamster ovary cells showed that diuron did not induce DNA cross-links. These data suggest that cytotoxicity with consequent regenerative cell proliferation is the predominant MOA for diuron rat urothelial carcinogenesis, the cytotoxicity being chemically induced and not due to urinary solids. PMID- 19812367 TI - Augmented bladder urothelial polyamine signaling and block of BK channel in the pathophysiology of overactive bladder syndrome. AB - Overactive bladder syndrome (OAB) is an idiopathic condition characterized by urinary urgency and urge incontinence. Detrusor overactivity has been traditionally described as the physiologic mechanism for OAB. However, the bladder urothelium (BU) may also be involved in the pathophysiology. This study measured polyamine signaling and its downstream effects on membrane conductivity in bladder urothelial cells (BUC) obtained from asymptomatic and OAB subjects. Immunohistofluorescence was used to measure ornithine decarboxylase (ODC) expression in BU. BUC, cultured from BU biopsies, were used for electrophysiologic studies. dl-alpha-Difluoromethylornithine (DFMO), spermine, or spermidine was used to modulate polyamine signaling in BUC. Results showed ODC overexpression in OAB BU. In OAB BUC, whole cell and cell-attached configuration showed significantly decreased currents. Using inside-out patches, outward currents increased significantly, suggesting a cytoplasmic source of the outward current block in OAB BUC. In control BUC, outward currents were mediated by the large-conductance calcium-activated potassium (BK) channel due to calcium dose dependence and block by iberiotoxin. Spermidine and spermine blocked the outward current in normal BUC in dose-dependent fashion. Conversely, DFMO significantly increased (P < 0.01) outward currents in OAB BUC both in cell-attached and in whole cell configuration. The outward currents in DFMO-treated-OAB BUC could be significantly reduced (P < 0.05) by adding back spermidine and spermine. These data suggest that polyamine signaling is upregulated in OAB urothelium and OAB BUC. Furthermore, polyamines in BUC block the BK channel. Targeting of bladder urothelial polyamine signaling may represent a novel approach for OAB treatment based on pathophysiologic mechanisms. PMID- 19812368 TI - Role of p47(phox) in regulating Cdc42GAP, vimentin, and contraction in smooth muscle cells. AB - Cdc42GAP (GTPase activating protein) has been shown to regulate smooth muscle contraction as well as cell motility, adhesion, proliferation, and apoptosis. We have recently shown that Cdc42GAP activity is suppressed in smooth muscle cells during contractile activation, which is reversed by inhibitors of reactive oxygen species (ROS). Because p47(phox), a regulatory subunit of NAD(P)H oxidase, has been implicated in smooth muscle signaling, we determined whether this subunit modulates Cdc42GAP activity in response to contractile stimulation. Transfection of smooth muscle cells with plasmids encoding short hairpin RNA (shRNA) against p47(phox), but not plasmids for luciferase shRNA, inhibited the expression of p47(phox). ROS production and the suppression of Cdc42GAP activity in response to stimulation with 5-hydroxytryptamine (5-HT) were attenuated in cells producing p47(phox) shRNA compared with cells producing luciferase shRNA. In contrast, the addition of hydrogen peroxide to p47(phox)-deficient cells suppressed the activity of Cdc42GAP. Furthermore, exposure to hydrogen peroxide led to a decrease in Cdc42GAP activity in an in vitro assay. Cdc42 activation, p21 activated kinase 1 (PAK1) phosphorylation at Thr-423 (an indication of PAK activation), and vimentin phosphorylation at Ser-56 in response to 5-HT activation were also attenuated in smooth muscle cells producing shRNA against p47(phox). The knockdown of p47(phox) inhibited smooth muscle contraction during stimulation with 5-HT but not hydrogen peroxide. These results suggest that the p47(phox) subunit of NAD(P)H oxidase may mediate the agonist-induced GAP suppression by controlling ROS generation in smooth muscle cells during agonist stimulation. p47(phox)-regulated GAP affects smooth muscle contraction likely through the Cdc42/PAK1/vimentin pathway. PMID- 19812369 TI - In vivo veritas: the power of in situ manipulation of cells in a living animal. Focus on "Expression of plasmid DNA in the salivary gland epithelium: novel approaches to study dynamic cellular processes in live animals". PMID- 19812370 TI - Gastrin induces the interaction between human mononuclear leukocytes and endothelial cells through the endothelial expression of P-selectin and VCAM-1. AB - Gastric mucosal inflammation is frequently associated with hypergastrinemia, and a correlation exists between the level of gastrin and degree of gastritis. We have previously observed that gastrin promotes leukocyte-endothelial interactions and contributes to Helicobacter-induced inflammation in the rat mesentery. In the present study, we aimed to evaluate a possible proinflammatory activity of gastrin in humans. The interaction between human leukocytes [U-937 cells, peripheral blood polymorphonuclear (PMN), and peripheral blood mononuclear (PBMC) cells] and human umbilical vein endothelial cells (HUVEC) was analyzed in static and dynamic conditions. The endothelial expression of adhesion molecules [P selectin, E-selectin, intercellular adhesion molecule-1, vascular cell adhesion molecule (VCAM)-1] was analyzed by flow cytometry and fluorescent microscopy screening. Gastrin increased the static adhesion of U-937 cells to HUVEC (1 h; 10(-9) M: 122 +/- 9%; 10(-8) M: 143 +/- 17%; 10(-7) M: 162 +/- 14% vs. control, all P < 0.05). Incubation of HUVEC with gastrin (4 h) also increased PBMC rolling (vehicle: 63 +/- 12; 10(-9) M: 109 +/- 29; 10(-8) M: 141 +/- 24; 10(-7) M: 261 +/ 16 leukocytes/min, P < 0.05) and adhesion (vehicle: 3 +/- 2, 10(-9) M: 11 +/- 4, 10(-8) M: 17 +/- 5, 10(-7) M: 15 +/- 5 leukocytes/mm(2), all P < 0.05) in the parallel-plate flow chamber. Treatment of PBMC with gastrin had no effects. The cholecystokinin (CCK)-2 receptor antagonist (L-365,260, 10(-7) M) prevented the effects of gastrin. P-selectin and VCAM-1 expression were enhanced by gastrin, and neutralizing antibodies against these molecules prevented PBMC rolling and adhesion. Gastrin did not affect the interactions between HUVEC and PMN. Gastrin induces interactions between human mononuclear leukocytes and endothelial cells through the activation of CCK-2 receptors and the enhancement of endothelial P selectin and VCAM-1. PMID- 19812371 TI - Topotecan triggers apoptosis in p53-deficient cells by forcing degradation of XIAP and survivin thereby activating caspase-3-mediated Bid cleavage. AB - The topoisomerase I inhibitor topotecan (TPT) is used in the therapy of different tumors including high-grade gliomas. We previously showed that TPT-induced apoptosis depends on p53 with p53 wild-type (wt) cells being more resistant because of p53-controlled degradation of topoisomerase I. Here, we show that p53 deficient (p53(-/-)) fibroblasts undergo excessive mitochondrial apoptosis featuring H2AX phosphorylation, Bcl-x(L) decline, cytochrome c release, caspase 9/-3/-2 activation, and cleavage of Bid. In wt and apaf-1(-/-) cells, caspase-2 did not become activated and Bid was not cleaved. In addition, p53(-/-) cells cotreated with TPT and caspase-3 inhibitor showed neither caspase-2 activation nor Bid cleavage, implying that caspase-2 is processed downstream of the apoptosome by caspase-3. Although processing of caspase-9/-3 was similar in wt and p53(-/-) cells, only p53(-/-) cells displayed active caspase-3. This was due to the proteasomal degradation of X-chromosome-linked inhibitor of apoptosis (XIAP) and survivin that inhibits caspase-3 activity. Accordingly, TPT-induced apoptosis in wt cells was increased after XIAP/survivin knockdown. Silencing of Bid led to reduction of TPT-triggered apoptosis. Data obtained with mouse fibroblasts could be extended to human glioma cells. In U87MG (p53wt) cells cotreated with TPT and pifithrin-alpha, or transfected with p53-siRNA, caspase-2 and Bid were significantly cleaved and XIAP/survivin was degraded. Furthermore, the knockdown of XIAP and survivin led to increased TPT-triggered apoptosis. Overall, the data show that p53-deficient/depleted cells are hypersensitive to TPT because they down-regulate XIAP and survivin, and thus amplify the intrinsic apoptotic pathway via caspase-3-mediated Bid cleavage. Therefore, in gliomas harboring wild-type p53, TPT-based therapy might be improved by targeted down regulation of XIAP and survivin. PMID- 19812372 TI - Control of oxidative phosphorylation by vitamin A illuminates a fundamental role in mitochondrial energy homoeostasis. AB - The physiology of two metabolites of vitamin A is understood in substantial detail: retinaldehyde functions as the universal chromophore in the vertebrate and invertebrate eye; retinoic acid regulates a set of vertebrate transcription factors, the retinoic acid receptor superfamily. The third member of this retinoid triumvirate is retinol. While functioning as the precursor of retinaldehyde and retinoic acid, a growing body of evidence suggests a far more fundamental role for retinol in signal transduction. Here we show that retinol is essential for the metabolic fitness of mitochondria. When cells were deprived of retinol, respiration and ATP synthesis defaulted to basal levels. They recovered to significantly higher energy output as soon as retinol was restored to physiological concentration, without the need for metabolic conversion to other retinoids. Retinol emerged as an essential cofactor of protein kinase Cdelta (PKCdelta), without which this enzyme failed to be activated in mitochondria. Furthermore, retinol needed to physically bind PKCdelta, because mutation of the retinol binding site rendered PKCdelta unresponsive to Rol, while retaining responsiveness to phorbol ester. The PKCdelta/retinol complex signaled the pyruvate dehydrogenase complex for enhanced flux of pyruvate into the Krebs cycle. The baseline response was reduced in vitamin A-deficient lecithin:retinol acyl transferase-knockout mice, but this was corrected within 3 h by intraperitoneal injection of vitamin A; this suggests that vitamin A is physiologically important. These results illuminate a hitherto unsuspected role of vitamin A in mitochondrial bioenergetics of mammals, acting as a nutritional sensor. As such, retinol is of fundamental importance for energy homeostasis. The data provide a mechanistic explanation to the nearly 100-yr-old question of why vitamin A deficiency causes so many pathologies that are independent of retinoic acid action. PMID- 19812373 TI - Novel role for the double-stranded RNA-activated protein kinase PKR: modulation of macrophage infection by the protozoan parasite Leishmania. AB - The evolution of Leishmania infection depends on the balance between microbicidal and suppressor macrophage functions. Double-stranded RNA (dsRNA)-activated protein kinase R (PKR), a classic antiviral protein, is able to regulate a number of signaling pathways and macrophage functions. We investigated the possible role of PKR in the modulation of Leishmania infection. Our data demonstrated that Leishmania amazonensis infection led to PKR activation and increased PKR levels. Consistently, in macrophages from PKR knockout 129Sv/Ev mice and RAW-264.7 cells stably expressing a dominant-negative (DN) construct of PKR (DN-PKR), L. amazonensis infection was strongly reduced. The treatment of infected macrophages with the synthetic double-stranded RNA poly(I:C), a potent PKR inductor, increased L. amazonensis intracellular proliferation. This effect was reversed by 2-aminopurine (2-AP), a pharmacological inhibitor of PKR, as well as by the expression of DN-PKR. NO release induced by dsRNA treatment was inhibited by L. amazonensis through NF-kappaB modulation. PKR activation induced by dsRNA also resulted in IL-10 production, whose neutralization with specific antibody completely abrogated L. amazonensis proliferation. Our data demonstrated a new role of PKR in protozoan parasitic infection through IL-10 modulation. PMID- 19812374 TI - The P2X7 purinergic receptor: from physiology to neurological disorders. AB - Purine nucleotides are well established as extracellular signaling molecules. P2X receptors are ATP-gated cation channels that mediate fast excitatory transmission in diverse regions of the brain and spinal cord. Several P2X receptor subtypes, including P2X(7), have the unusual property of changing their ion selectivity during prolonged exposure to ATP, which results in progressive dilation of the channel pore and the development of permeability to molecules as large as 900 Da. The P2X(7) receptor was originally described in cells of hematopoietic origin, including macrophages, microglia, and certain lymphocytes, and mediates the influx of Ca(2+) and Na(+) ions, as well as the release of proinflammatory cytokines. P2X(7) receptors may affect neuronal cell death through their ability to regulate the processing and release of interleukin-1beta, a key mediator in neurodegeneration, chronic inflammation, and chronic pain. Activation of P2X(7) receptors provides an inflammatory stimulus, and P2X(7) receptor-deficient mice have substantially attenuated inflammatory responses, including models of neuropathic and chronic inflammatory pain. Moreover, P2X(7) receptor activity, by regulating the release of proinflammatory cytokines, may be involved in the pathophysiology of depression. The P2X(7) receptor may thus represent a critical communication link between the nervous and immune systems, while providing a target for therapeutic exploitation. This review discusses the current biology and cellular signaling pathways of P2X(7) receptor function, as well as insights into the role for this receptor in neurological/psychiatric diseases, outstanding questions, and the therapeutic potential of P2X(7) receptor antagonism. PMID- 19812375 TI - Identification of novel cell migration-promoting genes by a functional genetic screen. AB - Here we describe the identification of novel cell migration-promoting genes based on an unbiased functional genetic screen in cultured cells. After the introduction of the retroviral mouse brain cDNA library into NIH3T3 mouse fibroblast cells, migration-promoted cells were selected by a 3-dimensional migration assay using cell culture inserts. After 5 rounds of enrichment, cDNAs were retrieved from the cells with a selected phenotype. Cell migration-promoting activity was confirmed by independent migration assays for the retrieved cDNAs, among which further investigation was focused on coiled-coil-helix-coiled-coil helix domain-containing protein 2 (chchd2). Whereas overexpression of chchd2 promoted cell migration, knockdown of endogenous chchd2 expression reduced cell migration. Chchd2-induced cell migration was associated with augmented formation of actin stress fibers and focal adhesion, which was mediated through Akt, RhoA/ROCK, and Jnk pathways. CHCHD2 protein directly interacted with hyaluronic acid-binding protein 1 (HABP1) that possessed migration-suppressing activity. Intracellular localization and further functional studies suggested that CHCHD2 and HABP1 may mutually regulate each other to balance cell migration. Thus, chchd2 is a novel cell migration determinant identified by an in vitro functional genetic selection strategy. The selection method can also be useful for the isolation of genes that give other phenotypes of interest. PMID- 19812376 TI - Epigenetics: methylation-associated repression of heparan sulfate 3-O sulfotransferase gene expression contributes to the invasive phenotype of H-EMC SS chondrosarcoma cells. AB - Heparan sulfate proteoglycans (HSPGs), strategically located at the cell-tissue organ interface, regulate major biological processes, including cell proliferation, migration, and adhesion. These vital functions are compromised in tumors, due, in part, to alterations in heparan sulfate (HS) expression and structure. How these modifications occur is largely unknown. Here, we investigated whether epigenetic abnormalities involving aberrant DNA methylation affect HS biosynthetic enzymes in cancer cells. Analysis of the methylation status of glycosyltransferase and sulfotransferase genes in H-HEMC-SS chondrosarcoma cells showed a typical hypermethylation profile of 3-OST sulfotransferase genes. Exposure of chondrosarcoma cells to 5-aza-2' deoxycytidine (5-Aza-dc), a DNA-methyltransferase inhibitor, up-regulated expression of 3-OST1, 3-OST2, and 3-OST3A mRNAs, indicating that aberrant methylation affects transcription of these genes. Furthermore, HS expression was restored on 5-Aza-dc treatment or reintroduction of 3-OST expression, as shown by indirect immunofluorescence microscopy and/or analysis of HS chains by anion exchange and gel-filtration chromatography. Notably, 5-Aza-dc treatment of HEMC cells or expression of 3-OST3A cDNA reduced their proliferative and invading properties and augmented adhesion of chondrosarcoma cells. These results provide the first evidence for specific epigenetic regulation of 3-OST genes resulting in altered HSPG sulfation and point to a defect of HS-3-O-sulfation as a factor in cancer progression. PMID- 19812378 TI - Neuroinflammation and Parkinson disease: the silent battleground. PMID- 19812379 TI - Pathologic intracellular signaling in childhood pilocytic astrocytomas. PMID- 19812380 TI - Cerebrovascular disease in childhood cancer survivors: A Children's Oncology Group Report. AB - BACKGROUND: Curative therapy for childhood cancer has dramatically improved over past decades. Therapeutic radiation has been instrumental in this success. Unfortunately, irradiation is associated with untoward effects, including stroke and other cerebrovascular disease (CVD). The Children's Oncology Group (COG) has developed guidelines for screening survivors at risk for persistent or late sequelae of cancer therapy. OBJECTIVES: This review summarizes the pathophysiology and relevant manifestations of radiation-induced CVD and outlines the specific patient groups at risk for early-onset stroke. The reader will be alerted to the availability of the COG recommendations for monitoring, and, when applicable, specific screening and treatment recommendations will be highlighted. METHODS: A multidisciplinary task force critically reviewed the existing literature and scored the evidence to establish the current COG guidelines for monitoring health of survivors treated with head and neck irradiation. RESULTS: Previous head and neck exposure to therapeutic radiation is associated with latent CVD and increased risk for stroke in some patient groups. Common manifestations of radiation-induced CVD includes steno-occlusive disease, moyamoya, aneurysm, mineralizing microangiopathy, vascular malformations, and strokelike migraines. CONCLUSION: Risk for stroke is increased in survivors of pediatric CNS tumors, Hodgkin lymphoma, and acute lymphoblastic leukemia who received radiation to the brain and/or neck. As the population of survivors ages, vigilance for stroke and cerebrovascular disease needs to continue based on specific exposures during curative cancer therapy. PMID- 19812381 TI - Primary radiotherapy showed favorable outcome in treating extranodal nasal-type NK/T-cell lymphoma in children and adolescents. AB - Extranodal nasal-type natural killer (NK)/T-cell lymphoma is rarely observed in children and adolescents. We aim to investigate the clinical features, prognosis, and treatment outcomes in these patients. Thirty-seven patients were reviewed. There were 19, 14, 2, and 2 patients with stage I, stage II, stage III, and stage IV diseases, respectively. Among the patients with stage I and II disease, 19 patients received initial radiotherapy with or without chemotherapy, and 14 patients received chemotherapy followed by radiotherapy. The 4 patients with stage III and IV disease received primary chemotherapy and radiation of the primary tumor. Children and adolescents with extranodal nasal-type NK/T-cell lymphoma usually presented with early-stage disease, high frequency of B symptoms, good performance, low-risk age-adjusted international prognostic index, and chemoresistance. The complete response rate after initial radiotherapy was 73.7%, which was significantly higher than the response rate after initial chemotherapy (16.7%; P = .002). The 5-year overall survival (OS) and progression free survival (PFS) rates for all the patients were 77.0% and 68.5%, respectively. The corresponding OS and PFS rates for patients with stage I and II disease were 77.6% and 72.3%, respectively. Children and adolescents with early stage extranodal nasal-type NK/T-cell lymphoma treated with primary radiotherapy had a favorable prognosis. PMID- 19812382 TI - Genetic variation in CXCR4 and risk of chronic lymphocytic leukemia. AB - A genome-wide linkage scan has provided evidence for a chronic lymphocytic leukemia (CLL) susceptibility locus at 2q21 to which the chemokine receptor CXCR4 gene maps. Recent data provide some evidence for common variation in CXCR4 according to the polymorphic variant rs2228014 defining CLL risk. To examine the role of genetic variation in CXCR4 on CLL risk, we screened 188 familial CLL cases and 213 controls for germline mutations in the coding regions of CXCR4 and genotyped rs2228014 in 1058 CLL cases and 1807 controls. No association between rs2228014 and risk of CLL was seen (P = .83). One truncating (W195X) and 2 missense mutations with possible functional consequences (V139I and G335S) were identified among 186 familial cases and 0 in 213 controls sequenced. Our analysis provides no evidence that common variation in CXCR4 defined by rs228014 influences the risk of CLL, but that functional coding mutations in CXCR4 may contribute to familial CLL. PMID- 19812383 TI - Allogeneic stem cell transplantation after reduced-intensity conditioning in patients with myelofibrosis: a prospective, multicenter study of the Chronic Leukemia Working Party of the European Group for Blood and Marrow Transplantation. AB - From 2002 to 2007, 103 patients with primary myelofibrosis or postessential thrombocythemia and polycythemia vera myelofibrosis and a median age of 55 years (range, 32-68 years) were included in a prospective multicenter phase 2 trial to determine efficacy of a busulfan (10 mg/kg)/fludarabine (180 mg/m(2))-based reduced-intensity conditioning regimen followed by allogeneic stem cell transplantation from related (n = 33) or unrelated donors (n = 70). All but 2 patients (2%) showed leukocyte and platelet engraftment after a median of 18 and 22 days, respectively. Acute graft-versus-host disease grade 2 to 4 occurred in 27% and chronic graft-versus-host disease in 43% of the patients. Cumulative incidence of nonrelapse mortality at 1 year was 16% (95% confidence interval, 9% 23%) and significantly lower for patients with a completely matched donor (12% vs 38%; P = .003). The cumulative incidence of relapse at 3 years was 22% (95% confidence interval, 13%-31%) and was influenced by Lille risk profile (low, 14%; intermediate, 22%; and high, 34%; P = .02). The estimated 5-year event-free and overall survival was 51% and 67%, respectively. In a multivariate analysis, age older than 55 years (hazard ratio = 2.70; P = .02) and human leukocyte antigen mismatched donor (hazard ratio = 3.04; P = .006) remained significant factors for survival. The study was registered at www.clinicaltrials.gov as #NCT 00599547. PMID- 19812384 TI - Induction of B-cell development in adult mice reveals the ability of bone marrow to produce B-1a cells. AB - To study B-cell development from bone marrow (BM), we generated recombination activating gene 1 (Rag1)-targeted mice lacking mature lymphocytes. B-cell development can be induced in such mice by B cell-specific restoration of a functional Rag1 transcription unit. Follicular and marginal zone B cells populated the spleen when Rag1 expression was permitted. Notably, the peritoneal cavity was dominated by bona fide B-1a cells, as judged by surface markers and functional properties. These BM-derived B-1a cells exhibited a polyclonal VDJ repertoire with substantial N nucleotide insertions. Nevertheless, physiologic frequencies of phosphatidylcholine-specific B cells were detected. Importantly, the BM of young and 5-month-old mice was indistinguishable with regard to the potential to generate B-1a cells. PMID- 19812385 TI - Oxidative modification of von Willebrand factor by neutrophil oxidants inhibits its cleavage by ADAMTS13. AB - Elevated plasma von Willebrand factor (VWF) and low ADAMTS13 activity have been reported in several inflammatory states, including sepsis and acute respiratory distress syndrome. One hallmark of inflammation is neutrophil activation and production of reactive oxygen species, including superoxide radical, hydrogen peroxide, and hypochlorous acid (HOCl). HOCl is produced from hydrogen peroxide and chloride ions through the action of myeloperoxidase. HOCl can oxidize methionine to methionine sulfoxide and tyrosine to chlorotyrosine. This is of interest because the ADAMTS13 cleavage site in VWF, the Tyr(1605)-Met(1606) peptide bond, contains both oxidation-prone residues. We hypothesized that HOCl would oxidize either or both of these residues and possibly inhibit ADAMTS13 mediated cleavage. We therefore treated ADAMTS13 substrates with HOCl and examined their oxidative modification by mass spectrometry. Met(1606) was oxidized to the sulfoxide in a concentration-dependent manner, with complete oxidation at 75muM HOCl, whereas only a miniscule percentage of Tyr(1605) was converted to chlorotyrosine. The oxidized substrates were cleaved much more slowly by ADAMTS13 than the nonoxidized substrates. A similar result was obtained with multimeric VWF. Taken together, these findings indicate that reactive oxygen species released by activated neutrophils have a prothrombotic effect, mediated in part by inhibition of VWF cleavage by ADAMTS13. PMID- 19812386 TI - Urokinase-type plasminogen activator increases hepatocyte growth factor activity required for skeletal muscle regeneration. AB - The plasminogen system plays a crucial role in the repair of a variety of tissues, including skeletal muscle. We hypothesized that urokinase-type plasminogen activator (uPA) promotes muscle regeneration by activating hepatocyte growth factor (HGF), which, in turn, stimulates proliferation of myoblasts required for regeneration. In our studies, levels of active HGF and phosphorylation of the HGF receptor c-met were increased after muscle injury in wild-type mice. Compared with wild-type animals, mice deficient in uPA (uPA(-/-)) had markedly reduced HGF levels and c-met activation after muscle damage. This reduced HGF activity in uPA(-/-) animals was associated with decreased cell proliferation, myoblast accumulation, and new muscle fiber formation. On the other hand, HGF activity was enhanced at early time points in PAI-1(-/-) mice compared with wild-type mice and the PAI-1(-/-) animals exhibited accelerated muscle fiber regeneration. Furthermore, administration of exogenous uPA rescued HGF levels and muscle regeneration in uPA(-/-) mice, and an HGF-blocking antibody reduced HGF activity and muscle regeneration in wild-type mice. We also found that uPA promotes myoblast proliferation in vitro through its proteolytic activity, and this process was inhibited by an HGF-blocking antibody. Together, our findings demonstrate that uPA promotes muscle regeneration through HGF activation and subsequent myoblast proliferation. PMID- 19812387 TI - The use of drugs and nutritional supplements in top-level track and field athletes. AB - BACKGROUND: High use of medication and nutritional supplements has been reported in several sports. PURPOSE: To document the use of prescribed medication and nutritional supplements in female and male junior, youth, and adult track and field athletes depending on their sports discipline. STUDY DESIGN: Descriptive epidemiology study. METHODS: Analysis of 3 887 doping control forms undertaken during 12 International Association of Athletics Federations World Championships and 1 out-of-competitions season in track and field. RESULTS: There were 6 523 nutritional supplements (1.7 per athlete) and 3 237 medications (0.8 per athlete) reported. Nonsteroidal anti-inflammatory drugs (NSAIDs; 0.27 per athlete, n = 884), respiratory drugs (0.21 per athlete, n = 682), and alternative analgesics (0.13, n = 423) were used most frequently. Medication use increased with age (0.33 to 0.87 per athlete) and decreased with increasing duration of the event (from sprints to endurance events; 1.0 to 0.63 per athlete). African and Asian track and field athletes reported using significantly fewer supplements (0.85 vs 1.93 per athlete) and medications (0.41 vs 0.96 per athlete) than athletes from other continents. The final ranking in the championships was unrelated to the quantity of reported medications or supplements taken. Compared with middle distance and long-distance runners, athletes in power and sprint disciplines reported using more NSAIDs, creatine, and amino acids, and fewer antimicrobial agents. CONCLUSION: The use of NSAIDs in track and field is less than that reported for team-sport events. However, nutritional supplements are used more than twice as often as they are in soccer and other multisport events; this inadvertently increases the risk of positive results of doping tests. CLINICAL RELEVANCE: It is essential that an evidence-based approach to the prescribing of medication and nutritional supplements is adopted to protect the athletes' health and prevent them from testing positive in doping controls. PMID- 19812388 TI - Selective disruption of ER{alpha} DNA-binding activity alters uterine responsiveness to estradiol. AB - In vitro models have been used to demonstrate that estrogen receptors (ERs) can regulate estrogen-responsive genes either by directly interacting with estrogen responsive element (ERE) DNA motifs or by interacting with other transcription factors such as AP1. In this study, we evaluated estrogen (E(2))-dependent uterine gene profiles by microarray in the KIKO mouse, an in vivo knock-in mouse model that lacks the DNA-binding function of ERalpha and is consequently restricted to non-ERE-mediated responses. The 2- or 24-h E(2)-mediated uterine gene responses were distinct in wild-type (WT), KIKO, and alphaERKO genotypes, indicating that unique sets of genes are regulated by ERE and non-ERE pathways. After 2 h E(2) treatment, 38% of the WT transcripts were also regulated in the KIKO, demonstrating that the tethered mechanism does operate in this in vivo model. Surprisingly, 1438 E(2)-regulated transcripts were unique in the KIKO mouse and were not seen in either WT or alphaERKO. Pathway analyses revealed that some canonical pathways, such as the Jak/Stat pathway, were affected in a similar manner by E(2) in WT and KIKO. In other cases, however, the WT and KIKO differed. One example is the Wnt/beta-catenin pathway; this pathway was impacted, but different members of the pathway were regulated by E(2) or were regulated in a different manner, consistent with differences in biological responses. In summary, this study provides a comprehensive analysis of uterine genes regulated by E(2) via ERE and non-ERE pathways. PMID- 19812389 TI - Proteomic analysis of phosphorylated nuclear proteins underscores novel roles for rapid actions of retinoic acid in the regulation of mRNA splicing and translation. AB - Retinoic acid (RA) signaling is mediated by the retinoic acid receptor (RAR), belonging to the nuclear hormone receptor superfamily. In addition to its classical transcriptional actions, RAR also mediates rapid transcription independent (nongenomic) actions, consisting in the activation of signal transduction pathways, as the phosphatidyl-inositol-3-kinase or the ERK MAPK signaling pathways. RA-induced rapid transcription-independent actions play a role in different physiological contexts. As an effort toward understanding the functions of those rapid actions on signaling elicited by RA, we have identified nuclear proteins the phosphorylation state of which is rapidly modified by RA treatment in neuroblastoma cells, using a proteomic approach. Our results show that RA treatment led to changes in the phosphorylation patterns in two families of proteins: 1) those related to chromatin dynamics in relation to transcriptional activation, and 2) those related to mRNA processing and, in particular, mRNA splicing. We show that treatment of neuroblastoma cells with RA leads to alteration of the regulation of pre-mRNA splicing and mRNA translation. Thus, our results underscore novel functions for the rapid signaling elicited by RAR in the regulation of mRNA processing. We conclude that RA activation of signaling pathways can indeed regulate mRNA processing as part of a cellular response orchestrated by the nuclear receptor RAR. PMID- 19812390 TI - Genomic and nongenomic cross talk between the gonadotropin-releasing hormone receptor and glucocorticoid receptor signaling pathways. AB - The GnRH receptor (GnRHR), a member of the G protein-coupled receptor family, is a central regulator of reproductive function in all vertebrates. The peptide hormone GnRH exerts its effects via binding to the GnRHR in pituitary gonadotropes. We investigated the mechanisms of regulation of transcription of the mGnRHR gene in the mouse pituitary gonadotrope L beta T2 cell line by GnRH and dexamethasone (dex). Reporter assays with transfected mGnRHR promoter show that both dex and GnRH increase transcription of the mGnRHR gene via an activating protein-1 (AP-1) site. Real-time PCR confirmed this on the endogenous mGnRHR gene, and small interfering RNA experiments revealed a requirement for the glucocorticoid receptor (GR) for both the dex and GnRH response. Chromatin immunoprecipitation (ChIP) and immunofluorescence assays provide evidence that both GnRH and dex up-regulate the GnRHR gene via nuclear translocation and interaction of the GR with the AP-1 region on the mGnRHR promoter. We show that GnRH activates the unliganded GR by rapid phosphorylation of the GR at Ser-234 in a GnRHR-dependent fashion to transactivate a GRE reporter gene in L beta T2 and COS-1 cells. Using kinase inhibitors, we established a direct link between GnRH induced protein kinase C and MAPK activation, leading to unliganded GR phosphorylation at Ser-234 and transactivation of the glucocorticoid response element. Furthermore, we show that GnRH and dex synergistically activate the endogenous GnRHR promoter in L beta T2 cells, via a mechanism involving steroid receptor coactivator-1 recruitment to the GnRHR AP-1 region. Our results suggest a novel mechanism of rapid nongenomic cross talk between the hypothalamic pituitary-gonadal and hypothalamic-pituitary-adrenal axes via GnRHR-dependent phosphorylation and activation of the unliganded GR in response to GnRH. PMID- 19812391 TI - Child mental health problems as risk factors for victimization. AB - The current study examines the effects of child internalizing and externalizing symptoms on increases in victimization over a 1-year period. Using longitudinal data from the Developmental Victimization Survey (DVS), analyses are based on a national probability sample of 1,467 children aged 2-17. Results indicate that children with high levels of co-occurring internalizing and externalizing symptoms were particularly likely to experience increased exposure to several forms of victimization, including peer victimization, maltreatment, and sexual victimization, controlling for earlier victimization and adversity. The relationship of symptoms to victimization exposure differed across developmental stage. Elementary school-age children with high levels of symptoms were especially vulnerable to victimization by peers, whereas distressed youth in early adolescence were particularly vulnerable to sexual victimization. Mental health problems in childhood and adolescence appear to represent important risk factors for increased victimization. Future interventions might consider targeting youth with co-occurring internalizing and externalizing symptoms during especially vulnerable developmental stages. PMID- 19812392 TI - How health care reform can benefit children and adolescents. PMID- 19812393 TI - Four health care reforms for 2009. PMID- 19812394 TI - American roulette--contaminated dietary supplements. PMID- 19812396 TI - Videos in clinical medicine. Bone marrow aspiration and biopsy. PMID- 19812395 TI - Proteomics analysis of the nucleolus in adenovirus-infected cells. AB - Adenoviruses replicate primarily in the host cell nucleus, and it is well established that adenovirus infection affects the structure and function of host cell nucleoli in addition to coding for a number of nucleolar targeted viral proteins. Here we used unbiased proteomics methods, including high throughput mass spectrometry coupled with stable isotope labeling by amino acids in cell culture (SILAC) and traditional two-dimensional gel electrophoresis, to identify quantitative changes in the protein composition of the nucleolus during adenovirus infection. Two-dimensional gel analysis revealed changes in six proteins. By contrast, SILAC-based approaches identified 351 proteins with 24 proteins showing at least a 2-fold change after infection. Of those, four were previously reported to have aberrant localization and/or functional relevance during adenovirus infection. In total, 15 proteins identified as changing in amount by proteomics methods were examined in infected cells using confocal microscopy. Eleven of these proteins showed altered patterns of localization in adenovirus-infected cells. Comparing our data with the effects of actinomycin D on the nucleolar proteome revealed that adenovirus infection apparently specifically targets a relatively small subset of nucleolar antigens at the time point examined. PMID- 19812397 TI - Images in clinical medicine. Pachyderma. PMID- 19812398 TI - Will the cost curve bend, even without reform? PMID- 19812400 TI - MicroRNA expression, survival, and response to interferon in liver cancer. AB - BACKGROUND: Hepatocellular carcinoma is a common and aggressive cancer that occurs mainly in men. We examined microRNA expression patterns, survival, and response to interferon alfa in both men and women with the disease. METHODS: We analyzed three independent cohorts that included a total of 455 patients with hepatocellular carcinoma who had undergone radical tumor resection between 1999 and 2003. MicroRNA-expression profiling was performed in a cohort of 241 patients with hepatocellular carcinoma to identify tumor-related microRNAs and determine their association with survival in men and women. In addition, to validate our findings, we used quantitative reverse-transcriptase-polymerase-chain-reaction assays to measure microRNAs and assess their association with survival and response to therapy with interferon alfa in 214 patients from two independent, prospective, randomized, controlled trials of adjuvant interferon therapy. RESULTS: In patients with hepatocellular carcinoma, the expression of miR-26a and miR-26b in nontumor liver tissue was higher in women than in men. Tumors had reduced levels of miR-26 expression, as compared with paired noncancerous tissues, which indicated that the level of miR-26 expression was also associated with hepatocellular carcinoma. Moreover, tumors with reduced miR-26 expression had a distinct transcriptomic pattern, and analyses of gene networks revealed that activation of signaling pathways between nuclear factor kappaB and interleukin-6 might play a role in tumor development. Patients whose tumors had low miR-26 expression had shorter overall survival but a better response to interferon therapy than did patients whose tumors had high expression of the microRNA. CONCLUSIONS: The expression patterns of microRNAs in liver tissue differ between men and women with hepatocellular carcinoma. The miR-26 expression status of such patients is associated with survival and response to adjuvant therapy with interferon alfa. PMID- 19812399 TI - Defibrillator implantation early after myocardial infarction. AB - BACKGROUND: The rate of death, including sudden cardiac death, is highest early after a myocardial infarction. Yet current guidelines do not recommend the use of an implantable cardioverter-defibrillator (ICD) within 40 days after a myocardial infarction for the prevention of sudden cardiac death. We tested the hypothesis that patients at increased risk who are treated early with an ICD will live longer than those who receive optimal medical therapy alone. METHODS: This randomized, prospective, open-label, investigator-initiated, multicenter trial registered 62,944 unselected patients with myocardial infarction. Of this total, 898 patients were enrolled 5 to 31 days after the event if they met certain clinical criteria: a reduced left ventricular ejection fraction (< or = 40%) and a heart rate of 90 or more beats per minute on the first available electrocardiogram (ECG) (criterion 1: 602 patients), nonsustained ventricular tachycardia (> or = 150 beats per minute) during Holter monitoring (criterion 2: 208 patients), or both criteria (88 patients). Of the 898 patients, 445 were randomly assigned to treatment with an ICD and 453 to medical therapy alone. RESULTS: During a mean follow-up of 37 months, 233 patients died: 116 patients in the ICD group and 117 patients in the control group. Overall mortality was not reduced in the ICD group (hazard ratio, 1.04; 95% confidence interval [CI], 0.81 to 1.35; P=0.78). There were fewer sudden cardiac deaths in the ICD group than in the control group (27 vs. 60; hazard ratio, 0.55; 95% CI, 0.31 to 1.00; P=0.049), but the number of nonsudden cardiac deaths was higher (68 vs. 39; hazard ratio, 1.92; 95% CI, 1.29 to 2.84; P=0.001). Hazard ratios were similar among the three groups of patients categorized according to the enrollment criteria they met (criterion 1, criterion 2, or both). CONCLUSIONS: Prophylactic ICD therapy did not reduce overall mortality among patients with acute myocardial infarction and clinical features that placed them at increased risk. (ClinicalTrials.gov number, NCT00157768.) PMID- 19812401 TI - A randomized trial of doxycycline for Mansonella perstans infection. AB - BACKGROUND: Mansonella perstans infection is common in areas of Africa where Wuchereria bancrofti, a causative agent of lymphatic filariasis, is endemic. M. perstans is refractory to standard antifilarial therapies. The recent discovery of bacterial endosymbionts (e.g., wolbachia) in most filarial species, including M. perstans, provides new therapeutic options for reducing microfilaremia. METHODS: In an open-label, randomized trial, we recruited subjects with M. perstans microfilaremia, with or without concomitant W. bancrofti infection, from four villages in Mali and randomly assigned them to receive doxycycline, at a dose of 200 mg daily for 6 weeks (106 subjects), or no treatment (110). At 6 months, subjects who were coinfected with W. bancrofti underwent a second random assignment, to treatment with a single dose of albendazole (400 mg) and ivermectin (150 microg per kilogram of body weight) or no treatment. Subjects were monitored daily during the first 6-week study period for adverse events. M. perstans and W. bancrofti microfilarial levels were assessed at 6, 12, and 36 months. RESULTS: At 12 months, 67 of 69 subjects who had received treatment with doxycycline only (97%) had no detectable M. perstans microfilariae per 60 microl of blood, as compared with 10 of 63 subjects who had received no treatment (16%) (relative risk, 6.18; 95% confidence interval, 3.63 to 11.89; P<0.001). At 36 months, M. perstans microfilaremia remained suppressed in 48 of 64 subjects who had received treatment with doxycycline only (75%), a finding that was consistent with a macrofilaricidal effect of doxycycline. Vomiting was more frequent in the doxycycline-treated group than in the untreated group (17% vs. 4%). CONCLUSIONS: These results are consistent with previous findings that M. perstans harbors the intracellular endosymbiont, wolbachia, and suggest that doxycycline is an effective therapy for M. perstans infection. (ClinicalTrials.gov number, NCT00340691.) PMID- 19812402 TI - Osteoporosis associated with neutralizing autoantibodies against osteoprotegerin. AB - Autoantibodies against osteoprotegerin, which block the inhibitory effect of osteoprotegerin on signaling by the receptor activator of nuclear factor (NF) kappaB (RANK), were identified in a man with celiac disease who presented with severe osteoporosis and high bone turnover. The osteoporosis did not respond to the treatment of his celiac disease but was completely reversed by bisphosphonate therapy. Autoantibodies against osteoprotegerin were detected in three additional patients with celiac disease. Such autoantibodies may be associated with the development of high-turnover osteoporosis, but whether autoantibodies against osteoprotegerin commonly contribute to the pathogenesis of osteoporosis in patients with celiac disease remains to be determined. PMID- 19812403 TI - Accuracy of conflict-of-interest disclosures reported by physicians. AB - BACKGROUND: The recent public reporting of payments made to physicians by manufacturers of orthopedic devices provides an opportunity to assess the accuracy of physicians' conflict-of-interest disclosures. METHODS: We analyzed the reports of payments made to physicians by five manufacturers of total hip and knee prostheses in 2007. For each payment recipient who was an author of a presentation or served as a committee member or board member at the 2008 annual meeting of the American Academy of Orthopaedic Surgeons, the disclosure statement was reviewed to determine whether the payment had been disclosed. To ascertain the reasons for nondisclosure, a survey was administered to physicians who had received payments that were not disclosed. RESULTS: The overall rate of disclosure was 71.2% (245 of 344 payments). For payments that were directly related to the topic of the presentation at the meeting, the rate was 79.3% (165 of 208); for payments that were indirectly related, the rate was 50.0% (16 of 32); and for payments that were unrelated, the rate was 49.2% (29 of 59) (P=0.008). In the multivariate analysis, payments were also more likely to have been disclosed if they exceeded $10,000 (P<0.001), were directed toward an individual physician rather than a company or organization (P=0.04), or included an in-kind component (P=0.002). Among the 36 physicians who responded to the survey regarding reasons for nondisclosure (response rate, 39.6%), the reasons most commonly given for nondisclosure were that the payment was unrelated to the topic of presentation at the annual meeting (38.9% of respondents) and that the physician had misunderstood the disclosure requirements (13.9%); 11.1% reported that the payment had been disclosed but was mistakenly omitted from the program. CONCLUSIONS: In this study of self-reported conflict-of-interest disclosure by physicians at a large annual meeting, the rate of disclosure was 79.3% for directly related payments and 50.0% for indirectly related payments. PMID- 19812404 TI - DNA damage, aging, and cancer. PMID- 19812405 TI - Images in clinical medicine. Staghorn calculus. PMID- 19812406 TI - Case records of the Massachusetts General Hospital. Case 31-2009. A 26-year-old man with abdominal distention and shock. PMID- 19812407 TI - The confirmatory trial in comparative-effectiveness research. PMID- 19812408 TI - Micromanaging cancer. PMID- 19812409 TI - Mansonella perstans--the importance of an endosymbiont. PMID- 19812410 TI - Gray-matter injury in multiple sclerosis. PMID- 19812411 TI - Routine early angioplasty after fibrinolysis. PMID- 19812412 TI - Rates of serious infection after medical abortion. PMID- 19812413 TI - Thrombomodulin in atypical hemolytic-uremic syndrome. PMID- 19812414 TI - Alcoholic hepatitis. PMID- 19812415 TI - Indigenous Australians and living kidney donation. PMID- 19812416 TI - Hospice access for individuals with dementia. AB - Involvement in a hospice program is important because it may allow individuals with dementia to delay or prevent institutionalization as well as provide psychosocial support for their families. Once used mostly by patients with a terminal cancer, now more than one half of the hospice patients have diagnoses other than cancer. Yet hospice is still underused for individuals dying with advanced dementia. We conducted a pilot study of hospice agencies to determine barriers and characteristics of dementia hospice enrollment. Using a mailed questionnaire and interview, we looked at demographics, accessibility, training, referral sources, and marketing. Our analysis divided the agencies based on dementia census and availability to non-Medicare eligible individuals. Results showed hospices having Bridge and Transition programs had on average 4 times higher Alzheimer's disease (AD) and dementia census than hospices without these programs. The highest rated barriers to hospice use for individuals with dementia were prognosis, education, and finance. PMID- 19812417 TI - Replication protein A stimulates the Werner syndrome protein branch migration activity. AB - Loss of the RecQ DNA helicase WRN protein causes Werner syndrome, in which patients exhibit features of premature aging and increased cancer. WRN deficiency induces cellular defects in DNA replication, mitotic homologous recombination (HR), and telomere stability. In addition to DNA unwinding activity, WRN also possesses exonuclease, strand annealing, and branch migration activities. The single strand binding proteins replication protein A (RPA) and telomere-specific POT1 specifically stimulate WRN DNA unwinding activity. To determine whether RPA and POT1 also modulate WRN branch migration activity, we examined biologically relevant mobile D-loops that mimic structures in HR strand invasion and at telomere ends. Both RPA and POT1 block WRN exonuclease digestion of the invading strand by loading on the strand. However, only RPA robustly stimulates WRN branch migration activity and increases the percentage of D-loops that are disrupted. Our results are consistent with cellular data that support RPA enhancement of branch migration during HR repair and, conversely, POT1 limitation of inappropriate recombination and branch migration at telomeric ends. This is, to our knowledge, the first evidence that RPA can stimulate branch migration activity. PMID- 19812418 TI - Global analysis of transcriptional regulation by poly(ADP-ribose) polymerase-1 and poly(ADP-ribose) glycohydrolase in MCF-7 human breast cancer cells. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) and poly(ADP-ribose) glycohydrolase (PARG) are enzymes that modify target proteins by the addition and removal, respectively, of ADP-ribose polymers. Although a role for PARP-1 in gene regulation has been well established, the role of PARG is less clear. To investigate how PARP-1 and PARG coordinately regulate global patterns of gene expression, we used short hairpin RNAs to stably knock down PARP-1 or PARG in MCF 7 cells followed by expression microarray analyses. Correlation analyses showed that the majority of genes affected by the knockdown of one factor were similarly affected by the knockdown of the other factor. The most robustly regulated common genes were enriched for stress-response and metabolic functions. In chromatin immunoprecipitation assays, PARP-1 and PARG localized to the promoters of positively and negatively regulated target genes. The levels of chromatin-bound PARG at a given promoter generally correlated with the levels of PARP-1 across the subset of promoters tested. For about half of the genes tested, the binding of PARP-1 at the promoter was dependent on the binding of PARG. Experiments using stable re-expression of short hairpin RNA-resistant catalytic mutants showed that PARP-1 and PARG enzymatic activities are required for some, but not all, target genes. Collectively, our results indicate that PARP-1 and PARG, nuclear enzymes with opposing enzymatic activities, localize to target promoters and act in a similar, rather than antagonistic, manner to regulate gene expression. PMID- 19812419 TI - Is there a need of extra fluoride in children? AB - The issues related to fluoridation of water or fortification of tooth paste with compounds of fluorides are controversial. Fluoride is stored mainly in the bones, where it increases the density and changes the internal architecture, makes it osteoporotic and more prone to fractures. Fluoride consumption by human beings increases the general cancer death rate, disrupts the synthesis of collagen and leads to the breakdown of collagen in bone, tendon, muscle, skin, cartilage, lungs, kidney and trachea, causing disruptive effect on various tissues in the body. It inhibits antibody formation, disturbs immune system and makes the child prone to malignancy. Fluoride has been categorized as a protoplasmic poison and any additional ingestion of fluoride by children is undesirable. PMID- 19812420 TI - Sedation challenges in the pediatric ICU: is dexmedetomidine the solution? PMID- 19812421 TI - Severity of illness scoring: one step closer to "the trenches". PMID- 19812422 TI - Introduction of Hib containing pentavalent vaccine in national immunization program of India: the concerns and the reality! PMID- 19812423 TI - Is there any benefit of supplementing infant milk formulae with long chain polyunsaturated fatty acids? PMID- 19812424 TI - Long chain polyunsaturated fatty acids and immunity in infants. AB - An infant is usually born with a deficient immune system, and the long chain polyunsaturated fatty acids (LC-PUFA) in breast milk plays an important role in the development and maturation of infant's immune system. This article reviews the role of LC-PUFA in breast milk in the development of immunity and prevention of atopic manifestations in infants. The review also attempts to assess the correct proportion of these nutrients that needs to be present in infant formulae for babies in whom breast milk is unavailable and formula milk is unavoidable. It was concluded that LC-PUFA plays a vital role in overall development of immunity in the infant. Clinicians should ensure that LC-PUFA are supplied to the term and preterm infant in the form of breastmilk or provided in right proportions in formula, if breast milk is unavailable. PMID- 19812425 TI - Cardiogenic shock with hypereosinophilic syndrome. AB - We report a child with hypereosinophilic syndrome who presented with cardiogenic shock. In addition, she had skin and joint involvement. The clinical condition improved and eosinophil counts normalized with steroid therapy. However, the skin lesions and hypereosinophilia relapsed on stopping the steroids. The child was subsequently maintained in remission on low dose prednisolone. PMID- 19812426 TI - Early myoclonic encephalopathy. AB - Early myoclonic encephalopathy (EME) is a rare malignant epileptic syndrome. The erratic myoclonus with or without focal motor seizures, onset before 3 months of age, and persistent suppression-burst pattern in electroencephalograph (EEG) are accepted as the diagnostic criteria for EME. We report an 11 month old infant with EME which was secondary to non-ketotic hyperglycinemia. PMID- 19812427 TI - Day 1 blood glucose and outcome in critically ill children. AB - We analyzed the association between the day 1 glucose values in 209 children admitted to the PICU and the outcome (mortality). 58 (27.7%) children had hyperglycemia while 18 (8.6%) children had hypoglycemia, on day 1 after admission. Hypoglycemia was associated with higher mortality. This is contrary to the prevalent view supporting the association of hyperglycemia with poor outcome in the critically ill children. PMID- 19812428 TI - Intravenous immune globulin for severe acute myocarditis in children. AB - We evaluated high-dose (2g/kg) intravenous immunoglobulin (IVIG) for severe acute myocarditis in 13 children and compared them for survival with 12 children with myocarditis treated with only conventional therapy. Baseline characteristics were similar between the two groups. Both groups had poor left ventricular ejection fraction (LVEF) on admission. The mortality rate was 8% in the IVIG treated children as compared to 46% in controls (P=0.04). Our study supports the use of IVIG in severe acute myocarditis in children. PMID- 19812429 TI - Statistical reporting in Indian Pediatrics. AB - We analyzed 45 original articles from Indian Pediatrics for appropriateness of the statistical methods. Appropriate statistical tests (93%), no use of obscure test and use of exact P value were the positive findings observed. Sample size was calculated in 24% and confidence interval in 13%. There is a need to generate awareness regarding confidence interval and sample size calculations. PMID- 19812430 TI - Bovine colostrum in those with immunodeficiency. PMID- 19812431 TI - Retinopathy of prematurity: future vision and challenges. PMID- 19812433 TI - RUTF: imported or indigenous. PMID- 19812435 TI - Host genetics influences on HIV type-1 disease. AB - HIV host genetic studies seek to describe as comprehensively as possible the effect of human genetic variation on the individual response to HIV type-1 (HIV 1) infection. Many associations between specific gene variants and HIV-1 disease outcomes have been reported over the past 15 years. Although most of them have yet to be confirmed or have been proven false-positives, the identification of several definitive genotype-phenotype associations has shed new light on HIV-1 pathogenesis. This review discusses these results in the context of the new genome-wide approaches that now make it possible to globally assess the influence of the host genome on HIV-1-related outcomes. PMID- 19812436 TI - Dengue vaccine development and dengue viral neutralization and enhancement assays. AB - Dengue fever is a major tropical infectious disease that affects 50-100 million people each year. Its complications, namely dengue haemorrhagic fever and dengue shock syndrome, disproportionately afflict children and young adults. The primary goal of several vaccines now in development is to elicit protective neutralizing antibody responses; however, the exact definition of such responses remain unclear. Here, we review briefly the historical aspects of dengue vaccine development and current candidate dengue vaccines, and discuss various laboratory assays for gauging the neutralizing antibody responses to infection or vaccination, or both. We conclude that modification of current neutralization assays is required to improve the correlation between neutralization end point determinations and protection against secondary heterotypic dengue infections. PMID- 19812437 TI - Surveillance for neuraminidase-inhibitor-resistant influenza viruses in Japan, 1996-2007. AB - BACKGROUND: High usage of the neuraminidase inhibitor (NAI) oseltamivir in Japan since 2003 led the Neuraminidase Inhibitor Susceptibility Network to assess the susceptibility of community isolates of influenza viruses to oseltamivir and zanamivir. METHODS: Isolates were tested by the enzyme inhibition assay and by neuraminidase (NA) sequence analysis. RESULTS: Among 1,141 A(H3N2) viruses and 171 type B viruses collected in Japan during the 2003-2004 season, 3 (0.3%) A(H3N2) isolates showed high 50% inhibitory concentrations (IC(50)) to oseltamivir. Each possessed a known resistance NA mutation at R292K or E119V. During the 2004-2005 season, no resistance was found among 567 influenza A(H3N2) or 60 A(H1N1) isolates, but 1 of 58 influenza B isolates had an NAI resistance mutation (D197N). Sequence analysis found that 4 (3%) of 132 A(H1N1) viruses from 2005-2006 had known NA resistance mutations (all H274Y), but no additional resistant isolates were detected from that or the subsequent 2006-2007 season. Concurrent testing of a selection of 500 influenza B viruses from 2000 to 2006 showed significant variations between seasons in both oseltamivir and zanamivir IC(50) values, but no persistent increases over this period. CONCLUSIONS: Our findings suggested possible low-level transmission of resistant variants from oseltamivir-treated patients in several seasons in Japan but no sustained reductions in NAI susceptibility or consistently increased frequency of detecting resistant variants for any strain or subtype, despite high levels of drug use. In particular, although oseltamivir-resistant A(H1N1) viruses with the H274Y mutation spread globally in 2007-2008, we found little evidence for increasing levels of resistant A(H1N1) variants in Japan in preceding years. PMID- 19812438 TI - Oxidant stress in HIV-infected women from the Women's Interagency HIV Study. AB - BACKGROUND: Oxidant stress contributes to the pathogenesis of multiple conditions and can be assessed by measuring plasma F(2)-isoprostane concentrations. We hypothesized that oxidant stress is associated with plasma homocysteine concentration and risk factors for atherosclerosis in HIV-infected women. METHODS: We measured plasma F(2)-isoprostane concentrations in a cross-sectional study of 249 HIV-infected women attending the Bronx (NY, USA) site of the Women's Interagency HIV Study and assessed associations with plasma homocysteine concentration and other metabolic parameters by linear regression. RESULTS: In multivariate analysis, hepatitis C virus (HCV) viraemia, waist circumference, homocysteine concentration and serum aspartate aminotransferase level were positively associated with log F(2)-isoprostane concentration (all P<0.005). There was a trend for an inverse association between log F(2)-isoprostane and CD4(+) T-cell percentage (P=0.06). Among women with HCV infection, the FIB-4 index, an indirect marker of liver fibrosis derived from routine laboratory tests, was positively associated with log F(2)-isoprostane concentration. CONCLUSIONS: In this cross-sectional study of HIV-infected women, plasma F(2) isoprostane concentration was positively associated with homocysteine concentration, as well as HCV infection, abdominal obesity and aspartate aminotransferase level. PMID- 19812439 TI - Virological and immunological responses to efavirenz or boosted lopinavir as first-line therapy for patients with HIV. AB - BACKGROUND: Efavirenz and lopinavir boosted with ritonavir are both recommended as first-line therapies for patients with HIV when combined with two nucleoside reverse transcriptase inhibitors. It is uncertain which therapy is more effective for patients starting therapy with an advanced infection. METHODS: We estimated the relative effect of these two therapies on rates of virological and immunological failure within the Swiss HIV Cohort Study and considered whether estimates depended on the CD4(+) T-cell count when starting therapy. We defined virological failure as either an incomplete virological response or viral rebound after viral suppression and immunological failure as failure to achieve an expected CD4(+) T-cell increase calculated from EuroSIDA statistics. RESULTS: Patients starting efavirenz (n=660) and lopinavir (n=541) were followed for a median of 4.5 and 3.1 years, respectively. Virological failure was less likely for patients on efavirenz, with the adjusted hazard ratio (95% confidence interval) of 0.63 (0.50-0.78) then multiplied by a factor of 1.00 (0.90-1.12) for each 100 cells/mm(3) decrease in CD4(+) T-cell count below the mean when starting therapy. Immunological failure was also less likely for patients on efavirenz, with the adjusted hazard ratio of 0.68 (0.51-0.91) then multiplied by a factor of 1.29 (1.14-1.46) for each 100 cells/mm(3) decrease in CD4(+) T-cell count below the mean when starting therapy. CONCLUSIONS: Virological failure is less likely with efavirenz regardless of the CD4(+) T-cell count when starting therapy. Immunological failure is also less likely with efavirenz; however, this advantage disappears if patients start therapy with a low CD4(+) T-cell count. PMID- 19812440 TI - Smoking motivations and quitting motivations among HIV-infected smokers. AB - BACKGROUND: The aim of this study was to examine smoking motivation and motivation to quit, and determinants of these motivations among HIV-infected cigarette smokers. METHODS: We conducted a 1-day cross-sectional survey on cigarette smoking in a representative sample of HIV-infected outpatients of French hospitals. A cluster analysis was used to characterize respondents' smoking motivation. A logistic regression was performed to study the factors associated with motivation to quit, including the clusters describing smoking motives. RESULTS: We found four clusters of smoking motivation. These included intellectual/emotional support (22% of respondents), automatic/stress relief (22%; characterized by heavy smoking and strong dependency); weight control (29%; characterized by frequent symptoms of fat accumulation because of antiretroviral therapy) and pleasure/conviviality (27%; corresponding to 'lighter' smokers). In the logistic regression model, among other significant covariates (cigarette consumption level, tobacco dependence and perceived risk of developing a smoking related disease), the automatic/stress relief cluster was negatively correlated to the motivation to quit (odds ratio 0.39), whereas the weight control cluster was strongly associated to this motivation (odds ratio 2.87). CONCLUSIONS: The diversity of HIV-infected smokers' profiles suggests that different types of anti tobacco measures should be implemented in this population, such as information campaigns on specific risks incurred by HIV-infected smokers, non-specific and comprehensive measures for those for whom smoking is combined with other difficulties, and alternative therapeutic solutions for those who smoke to deal with lipodystrophy. PMID- 19812441 TI - The course of inactive hepatitis B in hepatitis-C-coinfected patients treated with interferon and ribavirin. AB - BACKGROUND: Interferon (IFN) combined with ribavirin (RBV) is an effective therapy for hepatitis C virus (HCV)-infected patients. Those who are coinfected with hepatitis B virus (HBV), however, might suffer from HBV reactivation. The aim of this study was to assess HBV reactivation in HCV-coinfected inactive HBV carriers following IFN/RBV. METHODS: A total of 32 HBV carriers with 4 log(10) copies/ml (range 5.2-6.5 log(10) copies/ml); however, these patients had no ALT flare either on treatment (n=2) or off treatment (n=1). During follow-up, 8 (36%) treated patients and 4 controls lost serum hepatitis B surface antigen (HBsAg; annual rate 6.5% versus 6.9%; P-value non-significant), whereas 4 and 2 patients seroconverted to antibodies against HBsAg, respectively. Hepatocellular carcinoma developed in 1 patient with SVR and 1 non-responder under nucleoside therapy for HBV, both with cirrhosis. No patient clinically decompensated. CONCLUSIONS: Inactive HBV carriers coinfected with HCV might achieve an SVR following IFN/RBV. Combination therapy carries a low risk of on- and off-treatment HBV reactivation and does not prevent HBsAg seroclearance. PMID- 19812442 TI - Stimulation of the interleukin-1 receptor and Toll-like receptor 2 inhibits hepatitis B virus replication in hepatoma cell lines in vitro. AB - BACKGROUND: Toll-like receptors (TLRs) are a key component of the innate immune system and TLR2 has been shown to be involved in the immunopathogenesis of hepatitis B virus (HBV) infection in vivo. We investigated the role of TLR2 stimulation of virus-infected hepatocyte cell lines as a potential antiviral mechanism in vitro. METHODS: The hepatoblastoma cell line HepG2 was transduced with recombinant HBV baculoviruses and the hepatoma cell line Huh-7 was transiently transfected with complimentary DNA clones of HBV. HBV viral replication was quantified after stimulation with interleukin (IL)-1beta and Pam 2-Cys, a synthetic TLR2 ligand, by measuring intracellular core-associated single stranded HBV DNA using Southern blot hybridization, as well as viral nucleocapsid formation using a non-denaturing immunoblot method. RESULTS: Stimulation of both cell lines in vitro with IL-1beta and Pam-2-Cys, both known to induce expression of the pro inflammatory cytokines tumour necrosis factor-alpha and IL-8 via a nuclear factor-kappaB dependent pathway, resulted in the inhibition of HBV DNA replication in the transduced HepG2 cells by up to 90% and nucleocapsid formation in the transiently transfected Huh-7 cells by up to 30%, when compared with mock treated cells. CONCLUSIONS: Hepatoma cell lines expressed functional IL-1 receptor and TLR2 receptors, which when stimulated led to a signalling cascade that inhibited HBV replication. These data support an active role for hepatocytes in inhibiting HBV replication and provide a rationale for the development of TLR agonists as potentially novel antiviral agents. PMID- 19812443 TI - Alpha-galactosylceramide in chronic hepatitis B infection: results from a randomized placebo-controlled Phase I/II trial. AB - BACKGROUND: The glycosphingolipid alpha-galactosylceramide (alpha-GalCer) is known to stimulate invariant natural killer T-cells (iNKTs) and is able to induce powerful antiviral immune responses. The present dose-escalating randomized placebo-controlled Phase I/II trial aimed to investigate antiviral activity and safety of alpha-GalCer as a novel class of treatment for chronic hepatitis B patients. METHODS: Patients were randomly assigned to 0.1 microg/kg (n=8), 1 microg/kg (n=6) or 10 microg/kg (n=6) alpha-GalCer or placebo (n=7) treatment. RESULTS: Almost all alpha-GalCer-treated patients showed a rapid and strong decrease in natural killer T-cell (NKT) numbers. Patients with high baseline NKT numbers showed immune activation, including natural killer cell activation, increased serum tumour necrosis factor-alpha and interleukin-6 levels, and development of fever. Three patients demonstrated a transient decrease in hepatitis B virus (HBV) DNA. Only one alpha-GalCer-treated patient had a sustained decrease in HBV DNA at the end of follow-up. Four patients discontinued therapy because of fever shortly after drug administration. No significant side effects were observed. CONCLUSIONS: alpha-GalCer (0.1-10 microg/kg) used as monotherapy for chronic hepatitis B infection resulted in a strong decrease of NKTs, but did not clearly affect HBV DNA and alanine aminotransferase levels. alpha-GalCer was poorly tolerated and is unlikely to be suitable as an alternative monotherapy to the current treatment regimen. PMID- 19812444 TI - Combined effect of C-reactive protein and stavudine on adipogenesis. AB - BACKGROUND: Subcutaneous fat wasting in HIV therapy is primarily associated with the use of stavudine (d4T) and zidovudine (AZT). We hypothesized that C-reactive protein (CRP) might have an additive effect on nucleoside reverse transcriptase inhibitor (NRTI)-mediated peripheral fat loss. METHODS: 3T3-F442A cells were exposed to AZT (6 microM), d4T (3 microM) and/or CRP (0.5 microg/ml) during differentiation. Differentiation was assessed by real-time PCR measurement of peroxisome proliferator-activated receptor (PPAR)gamma and CCAAT/enhancer-binding protein (C/EBP)alpha, by quantification of triglyceride accumulation and by determination of adiponectin expression and secretion. In addition, parameters of lipid accumulation, lipolysis, cell viability and apoptosis were examined. RESULTS: When preadipocytes were induced to differentiate in the presence of only AZT, d4T or CRP, only AZT significantly impaired adipogenic differentiation. When combined, d4T+CRP also led to reduced triacylglycerol accumulation, an effect not explained by CRP-induced apoptosis or cell death, but instead confirmed by reduced PPARgamma and C/EBPalpha expression and decreased expression of factors involved in lipogenesis, such as fatty acid synthase and acetyl-coenzyme A carboxylase. We observed further reduction in adiponectin expression and secretion when adipocytes were differentiated in the presence of AZT or d4T together with CRP. Addition of rosiglitazone (1 microM) had no effect on reduced adipogenesis, but partially rescued the effects of d4T and d4T+CRP on adiponectin production. CONCLUSIONS: We conclude that CRP at levels circulating in patients with HIV infection might promote the anti-adipogenic potential of d4T, a cooperative effect that could account for the in vivo observed variability in the development of lipoatrophy. PMID- 19812445 TI - Pharmacokinetics, safety and tolerability of a single oral dose of maraviroc in HIV-negative subjects with mild and moderate hepatic impairment. AB - BACKGROUND: Maraviroc is the first CCR5 antagonist and only oral entry inhibitor approved for the treatment of HIV type-1 infection. Maraviroc is extensively metabolized, primarily by cytochrome P450 3A4 and hence its pharmacokinetics might be affected by impaired hepatic function. The objective of this study was to evaluate the pharmacokinetics of maraviroc in subjects with mild or moderate hepatic impairment compared with subjects with normal hepatic function. Safety and tolerability were also assessed. METHODS: This was an open-label, non randomized, single-centre, parallel-group study. A total of 24 subjects with mild (n=8) or moderate (n=8) hepatic impairment, or normal hepatic function (n=8) received a single dose of 300 mg maraviroc. RESULTS: Relative to those with normal hepatic function, the geometric mean ratio (90% confidence interval) for the maximum observed plasma concentration (C(max)) of maraviroc was 111% (74.6 166) and 132% (89.6-194) for those with mild and moderate hepatic impairment, respectively; the area under the concentration-time curve from time 0 to the last quantifiable concentration (AUC(last)) was 125% (84.7-185) and 146% (100-212); oral clearance was 89% (53.2-150) and 83% (49.2-139); and renal clearance was 94% (70.5-126) and 131% (98.6-173), respectively. Maraviroc was well tolerated in all subjects. CONCLUSIONS: Although differences in maraviroc pharmacokinetics were noted in subjects with hepatic impairment compared with those with normal hepatic function, these do not currently support a dose modification. The single 300 mg dose of maraviroc was well tolerated by subjects with normal and impaired hepatic function. PMID- 19812446 TI - Insulin resistance impairs sustained virological response rate to pegylated interferon plus ribavirin in HIV-hepatitis C virus-coinfected patients: HOMAVIC ANRS HC02 Study. AB - BACKGROUND: The aim of this study was to assess the effect of insulin resistance (IR) on the response to hepatitis C virus (HCV) therapy in HIV-HCV-coinfected patients. METHODS: A total of 238 HIV-HCV-coinfected patients (74% male, mean +/ sd age 40 +/-5 years, mean alcohol intake <50 g/day and 38% HCV genotype 2 or 3), treated by standard or pegylated interferon-alpha2b plus ribavirin during 48 weeks were studied. Liver biopsies were assessed before treatment. Patients were considered to have IR when the homeostasis model assessment of IR (HOMA-IR) was >2.5. Multiple logistic regression with stepwise selection was used to estimate independent factors associated with sustained virological response (SVR). RESULTS: IR was present in 32% and significant liver fibrosis (Metavir>or=F2) in 74% of patients. Patients with SVR (96/238 [40%]) were more likely to be infected with HCV genotype 2 or 3 (54% versus 27%; P<0.0001), and had more severe liver fibrosis (>or=F3; 45% versus 30%; P=0.03). By multivariate analysis, a HOMA IR>2.5 had a negative effect on the SVR (odds ratio 0.49 [95% confidence interval 0.26-0.92]; P=0.05). CONCLUSIONS: A high HOMA-IR level is frequently found in HIV HCV-coinfected patients and is associated with a reduced SVR rate. Improving insulin sensitivity might be a useful adjunct to HCV therapy in HIV-HCV coinfected patients. PMID- 19812447 TI - Genetic analysis and putative role in resistance to antivirals of the human cytomegalovirus DNA polymerase UL44 processivity factor. AB - BACKGROUND: The human cytomegalovirus (HCMV) DNA polymerase is composed of the UL54 catalytic subunit and the UL44 accessory protein. UL44 increases the processivity of polymerase along the DNA template during replication and, incidentally, is a substrate for the UL97 phosphotransferase. The molecular mechanisms of HCMV resistance to antiviral drugs interfering with viral DNA synthesis reported so far only rely on the presence of amino acid changes within the UL97 and UL54 viral enzymes. We aimed to describe the natural polymorphism of UL44 and to analyse the changes of its amino acids potentially associated with HCMV resistance to antivirals. METHODS: The full-length UL44 gene sequence was compared to that of four reference strains (including the AD169 strain) and 43 clinical strains from patients who had not received any previous anti-HCMV treatment, and 25 blood samples from 15 HCMV-infected patients experiencing therapeutic failure and exhibiting genotypic traits of HCMV resistance to antivirals. RESULTS: Overall, seven different amino acid changes associated with natural polymorphisms were identified among the 433 residues of the UL44 protein, occurring at a frequency of 2.1% for five of them and 10.6% for the double change G296S+L319I. The analysis of the HCMV strains exhibiting genotypic resistance to antivirals did not show any changes in UL44 that had significant association with resistance mutations of UL97 and/or UL54. CONCLUSIONS: UL44 processivity factor exhibits a very low polymorphism that does not concern the assumed functional domains of the protein. From this preliminary study, UL44 does not seem to be involved in HCMV resistance to antivirals. PMID- 19812448 TI - Statin therapy and changes in hip circumference among HIV-infected participants in the ALLRT Cohort. AB - BACKGROUND: We aimed to determine whether statin exposure in antiretroviral treated individuals is associated with increases in hip circumference compared with HIV treatment without concomitant statin use. METHODS: This was a prospective multicentre cohort study involving individuals who had received antiretroviral therapy for at least 40 weeks and who were enrolled in the AIDS Clinical Trials Group Longitudinal Linked Randomized Trials cohort. There were 2,223 participants in the statin-unexposed group and 371 in the statin-exposed group. The main outcome measure was change in hip circumference at week 32. RESULTS: The 32-week change in hip circumference in the statin-exposed group was 0.60 cm greater (95% confidence interval 0.11-1.10; P=0.02) than in the statin unexposed group after adjustment for age, gender, race, baseline body mass index and thymidine analogue exposure. CONCLUSIONS: Our findings support the hypothesis that statins might be beneficial in lipoatrophy. Given the limited treatment options for this important problem, further studies are needed to confirm this effect and to determine its clinical significance. PMID- 19812449 TI - Efficacy and safety of darunavir/ritonavir in treatment-experienced HIV type-1 patients in the POWER 1, 2 and 3 trials at week 96. AB - BACKGROUND: Long-term (96-week) efficacy and safety of the protease inhibitor (PI) darunavir coadministered with low-dose ritonavir (DRV/r) was evaluated in HIV type-1 (HIV-1)-infected patients with extensive prior treatment experience in the POWER 1, 2 and 3 trials. METHODS: Patients with HIV-1 RNA>or=1,000 copies/ml and >or=1 primary PI mutation were randomized to receive either DRV/r 600/100 mg twice daily plus an optimized background regimen (OBR), or an investigator selected control PI (CPI) plus OBR (POWER 3 was a DRV/r 600/100 mg twice daily single-arm study). The proportion of patients with HIV-1 RNA<50 copies/ml at week 96 was assessed (intent-to-treat [ITT], time-to-loss of virological response algorithm). RESULTS: In total, 467 patients received DRV/r 600/100 mg twice daily; 124 patients received CPI(s). At week 96, 39% of DRV/r patients in POWER 1 and 2 (pooled analysis) versus 9% of CPI patients achieved HIV-1 RNA<50 copies/ml (ITT, time-to-loss of virological response algorithm; P<0.001). A similar proportion of DRV/r patients (42%) in POWER 3 achieved HIV-1 RNA<50 copies/ml at week 96. Mean absolute CD4(+) T-cell count increase for DRV/r at 96 weeks was 133 cells/mm(3) in POWER 1 and 2 and 103 cells/mm(3) in POWER 3. Grade 2-4 treatment emergent adverse events at least possibly related to DRV/r (>or=2% incidence, excluding laboratory abnormalities) were diarrhoea (3%), vomiting (3%), nausea (2%) and headache (2%). CONCLUSIONS: Treatment with DRV/r 600/100 mg twice daily was well tolerated and led to sustained virological and immunological responses in treatment-experienced HIV-1-infected patients over 96 weeks. PMID- 19812450 TI - Lower limb high arterial flow induced by tenofovir and emtricitabine treatment. AB - Here, we describe a case of an HIV-infected patient with right lower limb oedema that appeared after initiation of tenofovir and emtricitabine treatment. The patient was fully investigated by serial heart and vessel echo-Doppler examination. Oedema of the lower limb was attributed to a transient drug-induced fivefold increase in peripheral artery flow, which was induced by a reduction in peripheral arterial resistance. The possible mechanisms of disease are discussed. PMID- 19812451 TI - Telbivudine in the treatment of chronic hepatitis B: experience in HIV type-1 infected patients naive for antiretroviral therapy. AB - BACKGROUND: Telbivudine is a potent inhibitor of hepatitis B virus (HBV) replication without anti-HIV type-1 (HIV-1) activity demonstrated in vitro; however, very few clinical data on HIV-1-infected patients are available at present. Because it represents a therapeutic option in HIV-1-HBV-coinfected patients who do not require antiretroviral therapy, we strictly monitored three HIV-1-HBV-coinfected patients treated with telbivudine monotherapy for chronic hepatitis B. METHODS: We performed molecular analysis of HBV DNA and of HIV-1 reverse transcriptase and protease RNA and DNA sequences in three HIV-1-HBV coinfected patients treated with telbivudine monotherapy. RESULTS: Despite a transient and deep reduction of HIV-1 RNA, observed in two of the three patients studied, no genotypic resistance mutations were detected on both HIV-1 and HBV viruses. CONCLUSIONS: Telbivudine therapy for 24 weeks showed a potent anti-HBV activity in HIV-1-positive, hepatitis B e antigen-positive patients with high HBV viraemia. No direct anti-HIV-1 activity of telbivudine was demonstrated and no genotypic resistance mutations to anti-HIV-1 drugs was found; however, the transient but deep reduction of HIV RNA, after telbivudine introduction, deserves further investigation and a strict monitoring of HIV-1 viraemia in HIV-1-infected patients on treatment with this drug. PMID- 19812452 TI - Two types of drug-resistant hepatitis B viral strains emerging alternately and their susceptibility to combination therapy with entecavir and adefovir. AB - The most serious problem of nucleoside/nucleotide analogue therapy for hepatitis B virus (HBV) infection is the emergence of drug-resistant mutant virus. Here, we describe a patient with chronic hepatitis B infection with a complex drug resistant mutant virus during sequential therapy with lamivudine (3TC), entecavir (ETV) and adefovir dipivoxil (ADV). The patient was a 52-year-old male with positive hepatitis B e antigen and high HBV DNA (>7.6 log(10) copies/ml). Initial 3TC monotherapy offered little benefit and 3TC resistance was established by the virus with rtA181T and not rtM204V/I. HBV DNA was reduced slightly by replacement with ETV monotherapy and was followed by virological breakthrough. At that time, rtA181T was undetectable and the virus with rtM204V and rtL180M became predominant. ETV resistance was established by an additional rtS202G mutation. Efficacy of subsequent combination therapy with ADV and 3TC was limited because of reappearance of the virus with rtA181T, which might confer cross-resistance to 3TC and ADV. Final combination therapy with ETV and ADV reduced HBV DNA to 3.7 log(10) copies/ml for 5 months, which was the most effective therapy for this patient. Thus, two kinds of mutant viruses (rtM204V-related and rtA181T-related) appeared alternately in this patient. Combination therapy with ETV and ADV might have been effective because these drugs share therapeutic roles, that is, ETV affects the rtA181T-related virus and ADV affects the rtM204V-related virus. This is the first report suggesting clinical significance of combination therapy with ETV and ADV for controlling replication of the complex drug-resistant mutant HBV. PMID- 19812453 TI - Defining asymmetry in heart rate variability signals using a Poincare plot. AB - The asymmetry in heart rate variability is a visibly obvious phenomenon in the Poincare plot of normal sinus rhythm. It shows the unevenness in the distribution of points above and below the line of identity, which indicates instantaneous changes in the beat to beat heart rate. The major limitation of the existing asymmetry definition is that it considers only the instantaneous changes in the beat to beat heart rate rather than the pattern (increase/decrease). In this paper, a novel definition of asymmetry is proposed considering the geometry of a 2D Poincare plot. Based on the proposed definition, traditional asymmetry indices -Guzik's index (GI), Porta's index (PI) and Ehlers' index (EI)--have been redefined. In order to compare the effectiveness of the new definition, all indices have been calculated for RR interval series of 54 subjects with normal sinus rhythm of 5 min and 30 min duration. The new definition resulted in a higher prevalence of normal subjects showing asymmetry in heart rate variability. PMID- 19812454 TI - Automated identification of peristaltic pressure waves in oesophageal manometry investigations using the rolling correlation technique. AB - We have implemented the technique of rolling correlation coefficient as proposed by Buttfield and Bolton (2005 Real time measurement of RR intervals using a digital signal processor J. Med. Eng. Technol. 29 8-13) for ECG R-wave detection in the detection and timing of oesophageal peristalsis. 43 sequential patients attending for oesophageal manometry were retrospectively reviewed. Two expert reviewers visually assessed each swallow for normality of peristaltic amplitude and propagation speed. Automatic assessment was performed using rolling correlation, maximum amplitude, threshold and maximum gradient techniques of identifying onset of peristalsis. Rolling correlation was comparable with the maximum amplitude technique at identifying peristaltic pressure waves visually identified as present. Rolling correlation was most effective at correctly identifying propagation velocity as normal (698 out of 845 normally propagating waves) and highest correlation with expert visual assessment of percentage abnormal propagation for each patient (R value 0.918). In a sub-group of 11 studies assessed as displaying normal motility, rolling correlation gave lowest variation of propagation speed and highest consistency with visual assessment. The rolling correlation technique is effective and accurate at identifying oesophageal peristalsis and characterizing peristaltic propagation in manometric studies even in the presence of abnormally weak peristalsis and other confounding pressure perturbations. PMID- 19812455 TI - Assessing the blood volume and heart rate responses during haemodialysis in fluid overloaded patients using support vector regression. AB - This study aims to assess the blood volume and heart rate (HR) responses during haemodialysis in fluid overloaded patients by a nonparametric nonlinear regression approach based on a support vector machine (SVM). Relative blood volume (RBV) and electrocardiogram (ECG) was recorded from 23 haemodynamically stable renal failure patients during regular haemodialysis. Modelling was performed on 18 fluid overloaded patients (fluid removal of >2 L). SVM-based regression was used to obtain the models of RBV change with time as well as the percentage change in HR with respect to RBV. Mean squared error (MSE) and goodness of fit (R(2)) were used for comparison among different kernel functions. The design parameters were estimated using a grid search approach and the selected models were validated by a k-fold cross-validation technique. For the model of HR versus RBV change, a radial basis function (RBF) kernel (MSE = 17.37 and R(2) = 0.932) gave the least MSE compared to linear (MSE = 25.97 and R(2) = 0.898) and polynomial (MSE = 18.18 and R(2)= 0.929). The MSE was significantly lower for training data set when using RBF kernel compared to other kernels (p < 0.01). The RBF kernel also provided a slightly better fit of RBV change with time (MSE = 1.12 and R(2) = 0.91) compared to a linear kernel (MSE = 1.46 and R(2) = 0.88). The modelled HR response was characterized by an initial drop and a subsequent rise during progressive reduction in RBV, which may be interpreted as the reflex response to a transition from central hypervolaemia to hypovolaemia. These modelled curves can be used as references to a controller that can be designed to regulate the haemodynamic variables to ensure the stability of patients undergoing haemodialysis. PMID- 19812457 TI - Application and comparison of dynamic models to assess impact of loading variations on performance of denitrification filters. AB - Dynamic models were developed using two software packages to evaluate performance of a denitrification filter pilot under variable flow and nitrate loading conditions. Two data periods, including one at constant rate hydraulic loading and one under diurnal flow conditions, were used to calibrate the models. Two additional test periods, one with high nitrate loadings and the second with peak hydraulic loadings and intermittent methanol usage were used to validate the models. The paper presents the results and compares the features of the two models. PMID- 19812458 TI - Cardiorespiratory fitness and preserved medial temporal lobe volume in Alzheimer disease. AB - Exercise and cardiorespiratory (CR) fitness may moderate age-related regional brain changes in nondemented (ND) older adults. The relationship of fitness to Alzheimer disease (AD)-related brain change is understudied, particularly in the hippocampus, which is disproportionately affected in early AD. The role of apolipoprotein E4 (apoE4) genotype in modulating this relationship is also unknown. ND (n=56) and early-stage AD patients (n=61) over the age of 65 years had magnetic resonance imaging and CR fitness assessments. Voxel-based morphometry techniques were used to identify AD-related atrophy. We analyzed the relationship of CR fitness with white and gray matter within groups, assessed fitness-related brain volume change in areas most affected by AD-related atrophy, and then analyzed differential fitness-brain relationships between apoE4 carriers. Atrophy was present in the medial temporal, temporal, and parietal cortices in patients with mild AD. There was a significant positive correlation of CR fitness with parietal and medial temporal volume in AD patients. ND patients did not have a significant relationship between brain volume and CR fitness in the global or small volume correction analyses. There was not a significant interaction for fitness x apoE4 genotype in either group. In early stage AD, CR fitness is associated with regional brain volumes in the medial temporal and parietal cortices suggesting that maintaining CR fitness may modify AD-related brain atrophy. PMID- 19812459 TI - Prevention of dementia by intensive vascular care (PreDIVA): a cluster-randomized trial in progress. AB - BACKGROUND AND PURPOSE: Cardiovascular risk factors are associated with an increased risk of dementia. Treatment of hypertension and hypercholesterolemia is associated with a decrease in incident dementia. Whether interventions aimed at cardiovascular risk factors in late life also reduce dementia risk is unknown. Here, we report the outline of a pragmatic study that will attempt to answer this question and we describe the prevalence of cardiovascular risk factors in the target population. METHODS: We designed a large cluster-randomized trial with a 6 year follow-up in 3700 elderly subjects (70 to 78 y) to assess whether nurse-led intensive vascular care in primary care decreases the incidence of dementia and reduces disability. Secondary outcome parameters are mortality, incidence of vascular events, and cognitive functioning. Intensive vascular care comprises treatment of hypertension, hypercholesterolemia, diabetes and reducing overweight, smoking cessation, and stimulating physical exercise. RESULTS: Baseline data of 1004 subjects show that 87% of the subjects have 1 or more cardiovascular risk factors and 44% have even 2 or more risk factors amenable to treatment. Seventy-nine percent of the subjects receiving antihypertensive medication still have a systolic pressure of >140 mm Hg. CONCLUSIONS: In this older age group, the very high percentage of elderly subjects with cardiovascular risk factors illustrates the large window of opportunity for therapies directed to lower the cardiovascular risk and potentially also the risk for dementia. PMID- 19812460 TI - Computer-based cognitive training for mild cognitive impairment: results from a pilot randomized, controlled trial. AB - We performed a pilot randomized, controlled trial of intensive, computer-based cognitive training in 47 subjects with mild cognitive impairment. The intervention group performed exercises specifically designed to improve auditory processing speed and accuracy for 100 min/d, 5 d/wk for 6 weeks; the control group performed more passive computer activities (reading, listening, visuospatial game) for similar amounts of time. Subjects had a mean age of 74 years and 60% were men; 77% successfully completed training. On our primary outcome, Repeatable Battery for Assessment of Neuropsychological Status total scores improved 0.36 standard deviations (SD) in the intervention group (P=0.097) compared with 0.03 SD in the control group (P=0.88) for a nonsignificant difference between the groups of 0.33 SD (P=0.26). On 12 secondary outcome measures, most differences between the groups were not statistically significant. However, we observed a pattern in which effect sizes for verbal learning and memory measures tended to favor the intervention group whereas effect sizes for language and visuospatial function measures tended to favor the control group, which raises the possibility that these training programs may have domain specific effects. We conclude that intensive, computer-based mental activity is feasible in subjects with mild cognitive impairment and that larger trials are warranted. PMID- 19812461 TI - An open-label study of memantine treatment in 3 subtypes of frontotemporal lobar degeneration. AB - There are currently no Food and Drug Administration-approved treatments for frontotemporal lobar degeneration (FTLD). The objectives of this study were to explore the tolerability of memantine treatment in FTLD and to monitor for possible effects on behavior, cognition, and function. Forty-three individuals who met clinical criteria for FTLD [21 with frontotemporal dementia (FTD), 13 with semantic dementia (SD), and 9 with progressive nonfluent aphasia (PA)] received 26 weeks of open-label treatment with memantine at a target dose of 20 mg daily. Concurrent treatment with acetylcholinesterase inhibitors was prohibited. Cognitive and functional outcome measures included the Mini Mental State Examination, Alzheimer's Disease Assessment Scale-Cognitive (ADAS-cog), clinical dementia rating-sum of boxes, Neuropsychiatric Inventory (NPI), Frontal Behavior Inventory, Executive Interview (EXIT25), Texas Functional Living Scale (TFLS), Geriatric Depression Scale, and Unified Parkinson's Disease Rating Scale motor scale. Most subjects were able to tolerate the target dose of memantine. A transient improvement was observed on the total NPI score primarily in the FTD group. Variable declines were observed on the ADAS-cog, EXIT25, Frontal Behavior Inventory, NPI, TFLS, and UPDRS scores. The FTD and SD groups declined on most of the cognitive and behavioral outcome measures, but remained stable on the UPDRS, whereas the progressive nonfluent aphasia group remained relatively stable on the ADAS-cog, NPI, and TFLS, but declined on the UPDRS. Memantine was well-tolerated in these subjects. Future placebo-controlled trials of memantine in FTLD are warranted and may have greater power to detect behavioral and cognitive effects if focused on the FTD and SD clinical syndromes. PMID- 19812462 TI - Bivariate heritability of total and regional brain volumes: the Framingham Study. AB - Heritability and genetic and environmental correlations of total and regional brain volumes were estimated from a large, generally healthy, community-based sample, to determine if there are common elements to the genetic influence of brain volumes and white matter hyperintensity (WMH) volume. There were 1538 Framingham Heart Study participants with brain volume measures from quantitative magnetic resonance imaging who were free of stroke and other neurologic disorders that might influence brain volumes and who were members of families with at least 2 Framingham Heart Study participants. Heritability was estimated using variance component methodology and adjusting for the components of the Framingham stroke risk profile. Genetic and environmental correlations between traits were obtained from bivariate analysis. Heritability estimates ranging from 0.46 to 0.60 were observed for total brain, WMH, hippocampal, temporal lobe, and lateral ventricular volumes. Moderate, yet significant, heritability was observed for the other measures. Bivariate analyses demonstrated that relationships between brain volume measures, except for WMH, reflected both moderate to strong shared genetic and shared environmental influences. This study confirms strong genetic effects on brain and WMH volumes. These data extend current knowledge by showing that these 2 different types of magnetic resonance imaging measures do not share underlying genetic or environmental influences. PMID- 19812463 TI - Association between BDNF Val66Met polymorphism and Alzheimer disease, dementia with Lewy bodies, and Pick disease. AB - A functional polymorphism of the brain-derived neurotrophic factor (BDNF Val66Met) has been reported to affect memory-related hippocampal activity. Apolipoprotein E (ApoE) gene polymorphism is known to be associated with Alzheimer disease (AD), dementia with Lewy bodies (DLB), and Pick disease (PiD). We tested the hypothesis that BDNF Val and ApoE epsilon4 alleles confer susceptibility to AD, DLB, and PiD. The study included 160 AD, 34 DLB patients, 38 autopsy-confirmed PiD, and 164 age-matched healthy control (HC) probands. The frequency of the BDNF Val allele was significantly higher in AD, but there were no statistical differences in the allele distribution in PiD or in DLB as compared with HC. The Val/Met genotype occurred with statistically significantly higher frequency in PiD than in HC. The ApoE epsilon4 allele was significantly overrepresented in all dementias as compared with HC. Genotypes containing both ApoE epsilon4 and BDNF Val alleles occurred more frequently in all investigated dementias than in HC. We suggest that the presence of the BDNF Val allele in itself and in combination with the ApoE epsilon4 allele can be risk factors for AD, and the results indicate a synergistic effect of the 2 polymorphisms on DLB and PiD risk. PMID- 19812464 TI - Correlation of clinical features with argyrophilic grains at autopsy. AB - Argyrophilic grains (AGs) are a pathologic feature found in association with neurodegenerative disease. Some have suggested that these features may occur as a distinctive condition. We reviewed 80 subjects from our tissue bank with pathologically confirmed AGs and identified their clinical features. We compared these subjects' features to the features of subjects with matched clinical diagnoses but without AGs. Subjects with AGs represented 21.7% of the entire autopsy sample from 1999 to 2005 (80 out of 367). Of Alzheimer disease (AD) subjects, 43 out of 233 had AGs (18.4% of AD subjects); 11 out of 42 Parkinson disease with dementia subjects had AGs (26.1% of Parkinson disease with dementia subjects); 2 out of 9 dementia with Lewy bodies subjects had AGs (22.2% of dementia with Lewy bodies subjects); 4 out of 15 mild cognitive impairment subjects had AGs (26.7% of mild cognitive impairment subjects); and 20 out of 68 cognitively normal subjects had AGs (29.4% of cognitively normal). Subjects with AGs tended to be older but only significantly so in AD. Many comorbid non neurologic health conditions were seen in cases of AGs without any single predilection emerging. AGs occur in approximately 22% of the entire autopsy cohort and are likely associated with advanced age. No distinctive antemortem clinical features were over represented in the AG cases. AGs can occur with or without neurodegenerative conditions and can occur in the absence of significant cognitive decline. AGs are not clearly associated with any single comorbid health condition. PMID- 19812465 TI - Differences in coated-platelet production between frontotemporal dementia and Alzheimer disease. AB - OBJECTIVE: Coated-platelets are a subset of platelets produced by dual-agonist activation with collagen and thrombin. These platelets retain full-length amyloid precursor protein on their surface and correlate inversely with disease severity in Alzheimer disease (AD). We have now investigated coated-platelet production and its relationship with disease severity in frontotemporal dementia (FTD) patients to determine whether our earlier observations were unique to AD. METHODS: Coated-platelet levels were assayed in 40 FTD, 40 AD patients, and 40 controls. Both patient groups were equally divided between mild-stage (Clinical Dementia Rating < or =1) and advanced stage dementia (Clinical Dementia Rating >1). RESULTS: Coated-platelet levels were not significantly different between patients with early-stage and advanced stage FTD (P=0.9), whereas early-stage AD patients had significantly higher levels than advanced stage AD (P<0.001). In addition, coated-platelet production was significantly elevated in early-stage AD versus early-stage FTD patients (P=0.01). CONCLUSIONS: In contrast to AD, there is no significant relationship between disease severity and coated-platelet levels in FTD. Differences in coated-platelet levels between early-stage AD and early-stage FTD patients warrant further investigation for potential clinical applications in helping to differentiate between these 2 disorders. PMID- 19812466 TI - Depression and plasma amyloid beta peptides in the elderly with and without the apolipoprotein E4 allele. AB - Depression associated with low plasma amyloid-beta peptide 42 (Abeta42) leading to a high ratio of Abeta40/Abeta42, a biomarker of Alzheimer disease (AD), may represent a unique depression subtype. The relationship between low plasma Abeta42 in depression and the major risk factor of AD, apolipoprotein E4 (ApoE4), is unknown. With the goal of clarifying this relationship, we analyzed 1060 homebound elders with ApoE characterization and depression status in a cross sectional study. Plasma Abeta40 and Abeta42 were measured, and cognition were evaluated. In the absence of the ApoE4 allele, depressed subjects had lower plasma Abeta42 [median (Q1, Q3): 17.1 (11.6, 27.8) vs. 20.2 (12.9, 32.9) pg/mL, P=0.006], a higher Abeta40/Abeta42 ratio [median (Q1, Q3): 7.1 (4.6, 11.3) vs. 6.9 (3.4, 9.7), P=0.03], and lower cognitive function (mean+/-SD of Mini-Mental State Examination: 24.5+/-3.1 vs. 25.5+/-3.3, P<0.0001) than those without depression. In contrast, these relationships were not observed in the presence of ApoE4. Instead, regardless the depression status ApoE4 carriers had lower plasma Abeta42 and a higher Abeta40/Abeta42 ratio than non-ApoE4 carriers. Using multivariate logistic regression, it was found that depression was not associated with ApoE4 allele, but with the interaction between plasma Abeta42 and ApoE4 (odds ratio=3.94, 95% confidence interval=1.50, 10.33, P=0.005), denoting low plasma Abeta42 in the absence of ApoE4. Both ApoE4 carriers and non-ApoE4 carriers with depression had lower Abeta42 and a higher Abeta40/Abeta42 ratio in plasma compared with non-ApoE4 carriers without depression in the homebound elderly. As a combination of low plasma Abeta42 and high plasma Abeta40 has been shown to increase the risk of AD in 2 large cohort studies, amyloid-associated depression shown in this study may suggest a risk factor of AD in the absence of ApoE4. PMID- 19812467 TI - Patterns of cerebral hypoperfusion in amnestic and dysexecutive MCI. AB - Although early studies on mild cognitive impairment (MCI) focused on memory dysfunction; more recent studies suggest that MCI is clinically heterogeneous. The objective of this study is to examine patterns of cerebral perfusion in anmestic (N=12) and nonamnestic (N=12) single-domain MCI patients from 4 a priori regions of interest: middle and superior frontal cortex, posterior cingulate, and precuneus, to compare them relative to healthy controls (N=12), and to correlate perfusion with neuropsychologic measures. Relative to controls, all MCI patients had hypoperfusion in the posterior cingulate, bilaterally. MCI patients with executive dysfunctions also showed hypoperfusion in bilateral middle frontal cortex and the left precuneus relative to controls and in the left middle frontal cortex, left posterior cingulate, and left precuneus relative to amnestic MCI patients. Perfusion in the posterior cingulate correlated positively with memory performance whereas perfusion in all 4 a priori regions of interest, predominantly on the left side, correlated with executive function performance. The finding that single-domain MCI patients with prominent deficits in different cognitive domains exhibited different patterns of hypoperfusion relative to controls supports the existence of distinct subgroups of MCI. These data further suggest that cognitive impairment in MCI is related to cerebral hypoperfusion. PMID- 19812468 TI - A comparison study of mild cognitive impairment with 3 memory tests among Chinese individuals. AB - OBJECTIVE: To examine whether 3 common memory tests differ statistically in terms of mild cognitive impairment (MCI) discrimination rates and conversion rates to Alzheimer disease. METHODS: A sample of 329 Chinese patients who consulted our memory clinic in Shanghai were tested using tasks including the auditory verbal learning test (AVLT), the logical memory (LM) test, the Rey-Osterrieth complex figure test, and other neuropsychologic tasks. One hundred and forty-nine of these patients were tested again using the identical tests 2 years later. The diagnose standard of probable Alzheimer disease is the same as the standard of the National Institute of Neurological and Communicative Diseases and Stroke Alzheimer Disease and Related Disorders Association. RESULTS: The results of the MCI discrimination rates are as follows: AVLT-II (51%) >AVLT-I (31%) > complex figure test-II (27%) >LM-I (21%)=LM-II (21%) (I-immediate recall; II-delayed recall). The MCI group categorized based on LM-II cutoff has a higher conversion rate per year (24%), but also a higher reversal rate and missed diagnosis rate, whereas the group based on AVLT-II cutoff has a lower conversion rate per year (12%), but also a lower reversal rate and missed diagnosis rate. CONCLUSIONS: The MCI discrimination rate and the conversion rate among different episodic memory tests are differ considerably. PMID- 19812469 TI - Characteristics and performance of a modified version of the ADCS-CGIC CIBIC+ for mild cognitive impairment clinical trials. AB - INTRODUCTION: The Alzheimer's Disease Cooperative Study-Clinical Global Impression of Change (ADCS-CGIC) was modified for use in mild cognitive impairment (MCI) trials and tested in the ADCS MCI randomized clinical trial of donepezil, vitamin E, and placebo. We assessed feasibility for its use by determining whether or not: (1) it distinguished a medication effect at 6 months and 12 months, (2) baseline demographic or clinical characteristics predicted change, (3) there was an association between MCI-CGIC and change in other clinical measures in order to evaluate external or concurrent validity. METHODS: We used a generalized estimating equations approach for ordinal outcome data to test the effects of treatment, baseline characteristics, and change in clinical measures on the MCI-CGIC over 12 months, and ordinal logistic regression to assess the association between MCI-CGIC and change in clinical measures at 6 months and 12 months. RESULTS: On the MCI-CGIC overall, 12.9% and 10.6% were rated as having improved, and 31.6% and 39.8% as having worsened over 6 months and 12 months, respectively. The MCI-CGIC did not distinguish the donepezil or vitamin E groups from placebo at 6 and 12 months treatment. Variables at screening or baseline that were associated with worse CGIC scores over 6 and 12 months included white race, greater years of education, worse depression, dementia severity rating, cognitive, and daily activities scores, and lower memory domain scores on a neuropsychological battery. Rate of worsening on the MCI-CGIC over 12 months was associated with change on the Alzheimer Disease Assessment Scale-cognitive and on executive function. Worsening at 6 months and 12 months, separately, were associated with the corresponding change in Alzheimer Disease Assessment Scale-cognitive, Activities of Daily Living, Beck Depression Inventory, Mini-Mental State Examination, Clinical Dementia Rating sum of boxes, memory, and executive function. CONCLUSIONS: Change detected by the MCI-CGIC was associated with baseline clinical severity and with change in clinical ratings over 6 and 12 months, supporting the validity of a CGIC approach in MCI. The effect size of the donepezil-placebo difference was similar to that of other outcomes at 12 months. About 40% of MCI patients were judged worse and about 11% improved, consistent with clinical experience and other ratings. PMID- 19812470 TI - Rate of decline in Alzheimer disease measured by a Dementia Severity Rating Scale. AB - OBJECTIVE: To measure clinically relevant change in Alzheimer disease (AD) using a family member-completed Dementia Severity Rating Scale (DSRS) questionnaire. BACKGROUND: Measuring rate of change provides important clinical information. Most neuropsychologic scores change nonlinearly, complicating their use as a predictor of change throughout the illness. METHODS: DSRS and Mini Mental State scores were prospectively collected on 702 patients with AD from first evaluation until they became too impaired to return to clinic. RESULTS: DSRS score increased an average of 4.48 points/y [95% confidence interval (CI): 4.14-4.82] throughout the entire range of severity. In contrast, the Mini Mental State declined an average of 2.15 points/y (95% CI: 1.85-2.46) during the first 2 years, accelerated to 3.83 points/y (95% CI: 3.28-4.38) during the subsequent 3 years, and then slowed to an annual decline of 1.63 points during the last 2 years (95% CI: 0.21-3.05). A younger age of symptom onset was associated with an increased rate of DSRS change (P=0.03). CONCLUSIONS: The DSRS provides a clinical measure of functional impairment in AD that increases about 4.48 points/y from the earliest symptomatic stage until patients become too severely impaired to return to clinic. PMID- 19812471 TI - Evaluation of the validity and reliability of the Alzheimer Disease-related Quality of Life Assessment Instrument. AB - The Alzheimer Disease-related Quality of Life (ADRQL) instrument was developed to assess health-related quality of life in people with AD using assessments from family caregivers or professional staff. Validity and reliability of the ADRQL in its original form and a revised version are assessed in a sample consisting of persons in 3 residential settings (community, assisted living, nursing home). The ADRQL exhibits good item-internal consistency (67.5% of items met 0.40 standard); high correlation of items to hypothesized scales (85% of items met criteria for 4 of 5 subscales and the overall instrument); a good range of scores (21.6 to 100 for total sample revised instrument); very low missing data; and internal consistency reliability coefficients exceeding the minimum reliability standard for group comparisons (0.86 for total scores; range of 0.56 to 0.83 for subscales). In a community sample, the ADRQL discriminates between groups and is responsive over time in hypothesized directions. Characteristics of caregiver raters (community sample) including demographics and self-rated health were largely unrelated to scores. The results support the use of the ADRQL to evaluate health-related quality of life in persons with AD across various care settings and various stages of the disease. Modest correlations with cognitive and functional disability levels suggest that the ADRQL provides insight into other important dimensions of life experience in persons with dementia. The revised version has improved measurement properties and is recommended for use over the original. PMID- 19812472 TI - Changes in the quality of life of people with dementia living in care homes. AB - Quality of Life (QoL) is now an established outcome measure for people with dementia. There is a need to understand if measures are sensitive to change and what factors are associated with change in QoL in dementia to develop interventions to improve QoL and identify who may be most likely to benefit. This study aimed to assess change in QoL in people living in 24-hour care homes using the Quality of Life-Alzheimer's Disease (QOL-AD) scale and investigated which clinical factors predicted changes in QoL in dementia. We used the QOL-AD scale to rate individual and staff perceptions of residents' QoL, for 238 people with dementia, recruited from 24 care homes in the United Kingdom. A follow-up interview was undertaken at 20 weeks. One hundred ninety-two (80.7%) of the 238 residents interviewed at baseline were followed up. A reduction in residents' QoL was predicted by lower baseline depression and anxiety symptoms, higher baseline QoL ratings, and an increase in depressive symptoms and cognitive deterioration at follow-up. Although QoL does not necessarily diminish as dementia progresses, it is strongly influenced by the person with dementia's mood. Improvement in cognition and mood may lead to increased QoL. PMID- 19812473 TI - Associations between microinfarcts and other macroscopic vascular findings on neuropathologic examination in 2 databases. AB - Clinicians may undervalue brain microinfarcts because they are defined by neuropathology and not seen on magnetic resonance imaging (MRI). We sought to identify what neuropathologic vascular findings-likely to be evident on brain MRI during life-would predict the presence of microinfarcts. We sought associations between such findings and microinfarcts in neuropathology databases from the National Alzheimer's Coordinating Center (NACC) and the Adult Changes in Thought (ACT) study. Considering only subjects 65 years or older at death, microinfarcts were evaluated in 6189 from NACC and 219 from ACT. Despite different definitions being used, microinfarcts were common in both studies (19.7% in NACC and 16.0% in ACT), and their frequency increased significantly with age. In NACC specimens, after controlling for age and sex in multivariable models, microinfarcts were strongly associated with macroinfarcts [odds ratio (OR): 4.4, 95% confidence interval (CI): 3.8, 5.0], leukoencephalopathy (OR: 2.6, 95% CI: 2.1, 3.3), and hemorrhages (OR: 2.0, 95% CI: 1.6, 2.6). Similarly in the ACT specimens, microinfarcts were strongly associated with macroinfarcts (OR: 2.9, 95% CI: 1.4, 6.3). These neuropathologic associations suggest that people whose cranial MRI shows macroinfarcts, hemorrhages, or leukoencephalopathy are more likely also to have microinfarcts. PMID- 19812474 TI - Parkinson disease with dementia: comparing patients with and without Alzheimer pathology. AB - Subjects with Parkinson disease (PD) frequently develop dementia with greater than one-third meeting neuropathologic diagnostic criteria for Alzheimer disease (AD). The objective is to identify clinical and neuropathologic differences between Parkinson disease with dementia (PDD) subjects, with and without coexistent AD pathology. Neuropathologic examination was available on subjects diagnosed by clinicopathologic criteria with PDD-AD (N=23) and PDD+AD (N=28). A small subset of subjects with PDD-AD and PDD+AD had received at least 1 standardized neuropsychologic assessment. PDD+AD subjects were significantly older at age of PD onset and death, progressed to onset of dementia in less time, and had a shorter duration of PD symptoms before the onset of dementia. Education, responsiveness of L-dopa and dopaminergic medications, presence of cognitive fluctuations and hallucinations, and mean Mini-Mental State Examination, Global Deterioration Scale, Functional Assessment Staging, and Unified Parkinson Disease Rating Scale scores did not differ significantly between the 2 groups. The PDD+AD group had significantly greater total plaques, neuritic plaques, total tangles, and Braak stages compared with PDD-AD. This study suggests that it is difficult to distinguish PDD+AD and PDD-AD on the basis of movement, clinical, and neuropsychologic assessment. PDD-AD and PDD+AD have similar degrees of dementia and approximately half of PDD subjects have enough AD pathology to attain a neuropathologic diagnosis of AD. PDD can develop in the absence of significant Alzheimer pathology. PMID- 19812475 TI - Sodium profiling, but not cool dialysate, increases the absolute plasma refill rate during hemodialysis. AB - Intradialytic hypotension is often caused by a discrepancy between ultrafiltration and plasma refilling. Increasing the plasma refill rate could therefore reduce intradialytic hypotension. We used a recently developed method to measure the effect of cool dialysate and sodium (Na) profiling on refill during hemodialysis (HD). Using a Gambro AK200 with blood volume (BV) sensor plus computer-guided external pump, a high ultrafiltration rate quickly induced a preset BV reduction. A software feedback mechanism subsequently adjusted the ultrafiltration rate continuously to maintain BV between very narrow preset boundaries. The continuously changing, software-generated ultrafiltration rate then quantitatively equalled refill. Absolute plasma refill rate was measured in six stable patients without intradialytic hypotension, undergoing HD without intervention, with cool dialysate (1 degrees C below core temperature), and with Na profiling (gradually declining from 150 to 140 mmol/l). Baseline refill rate was 20.1 + or - 4.0 ml/min (mean + or - SD). Although cool dialysate did not affect refill (22.2 + or - 4.1 ml/min, p = 0.27 vs. baseline), Na profiling induced a significant improvement (26.8 + or - 3.7 ml/min, p = 0.006 vs. baseline). Using our method to measure absolute plasma refill rate during HD, we demonstrated that Na profiling indeed improves the plasma refill rate. A potential effect of cool dialysate could not be established. PMID- 19812476 TI - Preliminary investigation of seeding mesenchymal stem cells on biodegradable scaffolds for vascular tissue engineering in vitro. AB - We used epsilon-caprolactone/L-lactide (PCLA) as a biodegradable scaffold and bone marrow (BM) mesenchymal stem cells (MSCs) as seeding cells for vascular tissue engineering: we expected MSCs to grow in the scaffolds in a bioreactor. The MSCs we used were from the BM of dogs, and vascular scaffolds were carried out on the electrospinning process of PCLA copolymers. MSCs expressed CD44 and CD105 but did not express CD34 or CD14 at an identical time point. Scaffolds were nontoxic to cells and were favorable for the growth and migration of MSCs. After culture in a bioreactor with mechanical stimulation, cells completely covered the surfaces of PCLA scaffolds and penetrated or infiltrated into the inside of the scaffold structure. PMID- 19812477 TI - Comparison of two different minimized extracorporeal circulation systems: hematological effects after coronary surgery. AB - Cardiopulmonary bypass induces hemolysis and activation of inflammatory and coagulation systems as a result of a combination of mechanical trauma and biological mechanisms. The aim of our study was to evaluate the performance of two different minimized extracorporeal circulation (ECC) systems and to compare their influence on blood components. From January 2003 to December 2008, 1,218 patients underwent coronary artery bypass grafting with minimized ECC. The PRECiSe system (41%) consists of a microporous capillary membrane oxygenator (MO) and a diagonal pump (DeltaStream DP2). The MECC system (59%) is composed of a polymethylpentene MO with a plasma-tight diffusion membrane and a centrifugal pump (RotaFlow). Serial blood samples were taken preoperatively (T0), on arrival to intensive care unit (T1), 6 hours postoperatively (T2), and at discharge (T3). Demographic data, intraoperative, and technical parameters were similar in both groups. At T1 and T2, the platelet count in the PRECiSe group was significantly lower than that in the MECC group (p < 0.01). Furthermore, at T1, levels of lactate dehydrogenase were significantly higher in the PRECiSe group (p < 0.05). In addition, postoperative blood loss was significantly higher using the PRECiSe system (p < 0.05). In conclusion, cardiac surgery with the MECC system is associated with less postoperative bleeding and improved blood cell preservation. PMID- 19812478 TI - Evaluation of optical propagation in blood for noninvasive detection of prethrombus blood condition. AB - This article evaluates the optical propagation to detect a "prethrombus" blood noninvasively. Thrombosis is still an inevitable issue in use of blood pumps, and it is required to predict thrombus formation as early as possible. We focused on the red blood cell (RBC) aggregation that is one of the features of thrombogenic process. First, by using a computer simulation, we calculated the optical propagations in blood for the RBC aggregation and nonaggregation blood. This simulation is based on the Monte-Carlo method and attempts to calculate the optical characteristics of the blood stochastically. In our simulation, the optical propagation with the RBC aggregation showed a different characteristic from that of the nonaggregation. Next, we examined the optical propagation in bovine blood with various activated whole blood clotting time (ACT). The blood mixed with sodium citrate was circulated by a blood pump. The ACT was adjusted between 1,000 and 50 seconds by controlling the ratio of calcium chloride solution to sodium citrate. We confirmed the RBC aggregation by using microscopic images and microthromboses in the pump directly. As a result, we evaluated that the change of the optical propagation has a correlation with thrombogenic process just as it was observed in our computer simulation. Our data indicate that the measurement of optical propagation can detect a prethrombous blood condition with RBC aggregation. Our study will help to establish optical technologies to detect prethrombous continuously and noninvasively. PMID- 19812479 TI - Primary in-series palliation of hypoplastic left heart syndrome with mechanical lung assist in neonatal pigs. AB - Less than optimal outcomes for hypoplastic left heart syndrome may be related to unstable in-parallel circulation. Mechanical lung assist by (1) superior vena cava to right atrium pump with oxygenation (oxygenator assist), or (2) superior vena cava to pulmonary artery pump (pump assist) may permit successful neonatal in-series palliation. Nineteen 15-day-old piglets underwent single ventricle and bidirectional cavopulmonary shunt (BCPC) creation without mechanical lung assist (n = 8), with pump assist (n = 5), or with oxygenator assist (n = 6). Baseline hemodynamic measurements were not different between groups. Median survival for the cavopulmonary shunt alone, pump assist, and oxygenator assist groups was 28, 180, and 180 minutes, respectively (p = 0.0006). No differences in arterial oxygen concentration or bicarbonate levels were detected. Arterial carbon dioxide (p < or = 0.007) was higher in the cavopulmonary shunt alone versus lung assist groups. Cavopulmonary shunt alone animals had decreased mean arterial pressure (p < 0.02) and cerebral perfusion pressure (p = 0.029) and elevated left atrial pressure compared with lung assist groups (p < 0.05). This data demonstrates creation of a novel translational neonatal BCPC model in which mechanical lung assist augments survival. Early BCPC death was related to poor ventricular function and an inability to ventilate, issues that were improved with both types of lung assist. PMID- 19812480 TI - Report on the ASAIO-IFAO congress, 55th annual meeting of the American Society for Artificial Internal Organs and third biennial congress of the International Federation for Artificial Organs, Dallas, Texas, May 28-30, 2009. PMID- 19812481 TI - Antimicrobial prophylaxis for primary immunodeficiencies. AB - PURPOSE OF REVIEW: Antibiotic prophylaxis is one of the mainstays of therapy of primary immunodeficiencies. We aim to summarize what is known about antibiotic prophylaxis for select primary immunodeficiencies. RECENT FINDINGS: In recent years, there has been a push towards more evidence-based practices for antimicrobial prophylaxis for many conditions such as antifungal prophylaxis for extremely premature neonates and antibiotic prophylaxis for neutropenia associated with chemotherapy. However, there are remarkably few data regarding antibiotic prophylaxis in primary immunodeficiencies and regimens vary greatly between practices. SUMMARY: Currently, antibiotic prophylaxis is guided by the common microbial pathogens seen in specific immunodeficiencies, and experience with other chronic illnesses such as cystic fibrosis, HIV, and immunosuppression from transplantation. Controlled studies are necessary to address the preferred antimicrobial and immunomodulator regimens for most of the primary immunodeficiencies. PMID- 19812482 TI - Certainties and doubts about sublingual and oral immunotherapy in children. AB - PURPOSE OF REVIEW: Sublingual (SLIT) and oral immunotherapy (OIT) are two variants of immunotherapy that might be especially of interest for children. After adult studies, lately many trials in children are published. However, the quality of published material varies. In the present review we analyze paediatric SLIT publications from the past 2 years in the light of a new system of evaluation of quality of evidence, the grading of recommendations assessment, development and evaluation system. RECENT FINDINGS: High quality evidence in children: high-dose daily grass SLIT reduces symptoms and medication use; low dose pollen SLIT reduces the development of new sensitizations. Low-dose SLIT reduces symptoms and medication in mild-moderate atopic dermatitis. Latex SLIT for 3 years results in a negative glove test. The evidence for the effectiveness of SLIT for seasonal and perennial asthma is still of moderate-low quality, as is the evidence for asthma prevention. Too high dose of mite SLIT seems ineffective for asthma. Oral allergen immunotherapy is promising for some food allergies. Some definite immunological changes are seen with SLIT; facilitated antibody presentation and IL-17 might correlate with efficacy. SUMMARY: Even after a hundred years of immunotherapy, it is still an ever changing field with SLIT and OIT making fast progress. PMID- 19812483 TI - Automated anesthesia. AB - PURPOSE OF REVIEW: Anesthesiologists are overloaded with information and multitasking necessities in an extremely complex work environment. The purpose of this review is to present recent developments toward automated anesthesia and present future technologies for everyday clinical practice. RECENT FINDINGS: Decision support systems integrate different parameters, clinical scenarios and assessments by (non)-trained personnel into algorithms, which lead to diagnostic suggestions, triage evaluations or treatment options. Target-controlled anesthesia infusion systems reduce the anesthesiologist's workload; target controlled analgesia systems have the potential to provide more stable hemodynamic control. Closed-loop delivery of anesthesia is feasible and provides anesthetic control as good as or better than human delivery. Teleanesthesia offers the possibility of distant preoperative assessment of the patient's fitness for anesthesia, aid of trained personnel to perform anesthetic tasks and the control of anesthesia delivery in a distant location. SUMMARY: Decision support systems help to make reliable and standardized decisions in complex environments. Target-controlled infusion systems reduce the anesthetic workload. Closed-loop systems will automate anesthesia care in the near future. Teleanesthesia offers the opportunity to provide safe anesthetic care whenever trained personnel are not available or need support. PMID- 19812484 TI - Ambulatory surgery and malignant hyperthermia. AB - PURPOSE OF REVIEW: It is timely to review recent publications that add to our knowledge of malignant hyperthermia because we can go to the web or to newspapers and read of recent tragic malignant hyperthermia deaths. RECENT FINDINGS: New methods were used to affirm some of the previous observations regarding the epidemiology of malignant hyperthermia, and the well known North American Malignant Hyperthermia Registry was used to describe characteristics of malignant hyperthermia deaths. Anesthesia workstations have been found to have a larger reservoir of potent inhalation anesthetics than did their predecessors. Activated charcoal can assist in cleaning these machines. Genetic diagnosis of malignant hyperthermia susceptibility can sometimes be obtained from blood specimens. Work continues to increase the sensitivity of blood-based testing of malignant hyperthermia susceptibility. SUMMARY: Because malignant hyperthermia is potentially lethal, families should be evaluated as thoroughly as possible when an individual who claims to be malignant hyperthermia susceptible presents for elective surgery. Genetic testing may facilitate the evaluation of families with a very strong history of malignant hyperthermia. The use of activated charcoal can speed the removal of potent inhalation anesthetics from anesthesia workstations. This should facilitate the anesthetic care of malignant hyperthermia susceptible patients. PMID- 19812485 TI - Cardiac surgery and acute kidney injury: emerging concepts. AB - PURPOSE OF REVIEW: To review recent advances in understanding of the occurrence, pathophysiology, prophylaxis and treatment of cardiac surgery-associated acute kidney injury (AKI). RECENT FINDINGS: New definitions and prompt diagnostic tools, enhanced risk stratification strategies, avoidance of nephrotoxins and procedure selection are all areas in which recent contributions to the literature have improved perioperative care for cardiac surgery patients. Although evidence continues to confirm the adverse effects of extreme hemodilution and transfusion as part of cardiac surgery, the previously believed advantages of tight perioperative glucose control for the kidney are being questioned in recent studies. Although very little evidence from randomized trials in cardiac surgery populations supports specific interventions to protect or prevent AKI, recent reports indicate reduced AKI associated with some procedural innovations, and the hope that promising findings for agents such as sodium bicarbonate that require further study may yield effective therapies. SUMMARY: The vexing problem of AKI following cardiac surgery is common and unsolved. Clinical strategies that stress avoidance rather than treatment remain the mainstay of effective management of patients at high renal risk. PMID- 19812486 TI - Nutritional assessment and support in acute kidney injury. AB - PURPOSE OF REVIEW: Acute kidney injury (AKI) in the ICU is associated with an increased risk of protein-energy wasting (PEW), a major negative prognostic factor. This review illustrates recently published data and guidelines concerning nutritional problems in AKI, pointing out complexities and peculiarities of the syndrome. RECENT FINDINGS: The main goals of nutritional support in AKI on renal replacement therapy (RRT) are to ensure the provision of adequate amounts of nutrients, to prevent PEW, to promote tissue reparation, to support the immune system, and possibly to reduce mortality. The enteral route should be preferred, even though parenteral nutrition is often required to target nutritional needs. Special attention should be paid both to the impact of RRT on macronutrient and micronutrient losses, and to the risk of complications. In fact, due to both the acute loss of the kidneys' homeostatic function, and the frequent need of RRT, patients with AKI are especially prone to hypoglycemia and hyperglycemia, hypertriglyceridemia, fluid balance alterations, electrolyte and acid-base derangements. SUMMARY: This review highlights the most recent concepts and recommendations for nutritional support in AKI, stressing the need for a close integration between adequate nutrition and RRT in this clinical condition, with the aim of carefully tailoring both therapies on patients' changing needs. Recent findings about the renoprotective role of some nutrients (glutamine, omega-3 fatty acids) are also discussed. PMID- 19812487 TI - Battlefield resuscitation. AB - PURPOSE OF REVIEW: To bring together in one review article, the most current and relevant evidence relating to military trauma resuscitation. RECENT FINDINGS: The main themes highlighted by this review are coagulopathy of trauma shock (CoTS), damage control resuscitation, haemostatic resuscitation, the management of massive transfusion, use of adjuvant drugs for haemostasis and use of an empiric massive transfusion protocol. SUMMARY: The review aims to educate the readership in recent advances in trauma practice, culminating in a novel empiric massive transfusion algorithm seamlessly guiding the clinician through the initial resuscitation stage resulting in reduced mortality, morbidity, coagulopathy and decreased overall blood product usage. PMID- 19812488 TI - Critical care organ support: a focus on extracorporeal systems. AB - PURPOSE OF REVIEW: An extensive search of the literature published in the past 2 years related to critical care organ support was undertaken. This review is limited to those that focus on extracorporeal life support modalities for adults. RECENT FINDINGS: Traditional indications for extracorporeal life support such as oxygenation, carbon dioxide exchange and support of perfusion have expanded to include solute and toxin clearance for kidney, liver and potentially neurological failure. Enhanced understanding of cell-mediated mechanisms of injury may explain multiple-organ dysfunction following single-organ damage. Extracorporeal life support systems can be used safely in patients with traumatic brain, chest, and abdominal injury. 'Emergency perfusion and resuscitation' following traumatic exsanguination is entering clinical trials. SUMMARY: Multiple-organ dysfunction followed by traumatic injury can be treated with multiple-organ support. A total extracorporeal organ support system may be used in the future as a portable, bedside organ support device. PMID- 19812489 TI - Nonmyeloablative conditioning with total lymphoid irradiation and antithymocyte globulin: an update. AB - PURPOSE OF REVIEW: The immune modulatory effects of total lymphoid irradiation (TLI) for graft-versus-host disease (GVHD) protection and transplantation tolerance following allogeneic bone marrow and organ transplantation have been studied for years in animal models. In preclinical models nonmyeloablative TLI conditioning alters residual host T cell subsets to favor regulatory natural killer T cells that suppress GVHD and prevent organ allograft rejection. These preclinical models have been recently adapted to human transplantation. RECENT FINDINGS: Patients receiving allogeneic hematopoietic cell transplantation for hematological malignancies conditioned with TLI and depletive T cell antibodies showed sustained donor chimerism, a reduced incidence of acute GVHD yet retained graft antitumor activity. As in the preclinical models, nonmyeloablative TLI conditioning significantly altered residual host T cell subsets favoring natural killer T cells, and the low incidence of GVHD was associated with increased IL-4 secretion by chimeric donor T cells. The TLI regimen used in cancer patients was modified to determine conditions for stable mixed chimerism and tolerance induction following combined hematopoietic cell and kidney transplantation. SUMMARY: This review summarizes the evolution of the preclinical TLI protocols and their recent translation to clinical trials, and discusses the mechanisms involved in protection from GVHD and the induction of tolerance following mixed chimerism. PMID- 19812490 TI - Acute graft-versus-host disease: new treatment strategies. AB - PURPOSE OF REVIEW: Graft-versus-host disease (GVHD) remains a major cause of morbidity and mortality after allogeneic hematopoietic cell transplantation (HCT), despite improvements in our understanding of its pathophysiology as well as the generation of new monoclonal antibodies, immunomodulatory chemotherapy, cellular therapeutics and supportive care. Herein, we review therapies that have proven effective as well as newer agents that have recently improved GVHD response rates and survival following HCT. RECENT FINDINGS: Novel approaches to prevent or treat GVHD are often based on evidence from experimental models. Our understanding of the pathophysiology of GVHD may lead to the development of innovative strategies that target both soluble and cellular effectors. Among such agents are sirolimus, anti-tumor necrosis factor antibodies, anti-LFA-3-IgG fusion protein, extracorporeal photopheresis, mesenchymal stem cells and regulatory T cells. SUMMARY: Obstacles to the improvement of HCT include the tight linkage between GVHD toxicity and the beneficial graft-versus-leukemia (GVL) effect, as well as the impairment of immune reconstitution by immunomodulatory drugs leading to life-threatening infections. The design of newer phase I/II clinical trials are underway. Future therapies are likely to include modulation of cell types that play key roles in the GVH process, including regulatory T cells, dendritic cells, natural killer T cells and B cells. PMID- 19812491 TI - Blood groups and malaria: fresh insights into pathogenesis and identification of targets for intervention. AB - PURPOSE OF REVIEW: This review summarizes recent advances in our understanding of the interaction between malaria parasites and blood group antigens and discusses how the knowledge gleaned can be used to target the development of new antimalarial treatments and vaccines. RECENT FINDINGS: Studies of the interaction between Plasmodium vivax and the Duffy antigen provide the clearest example of the potential for basic research on blood groups and malaria to be translated into a vaccine that could have a major impact on global health. Progress is also being made in understanding the effects of other blood group antigens on malaria. After years of controversy, the effect of ABO blood groups on falciparum malaria has been clarified, with the non-O blood groups emerging as significant risk factors for life-threatening malaria, through the mechanism of enhanced rosette formation. The Knops blood group system may also influence malaria susceptibility, although conflicting results from different countries mean that further research is required. Unanswered questions remain about the interactions between malaria parasites and other blood group antigens, including the Gerbich, MNS and Rhesus systems. SUMMARY: The interplay between malaria parasites and blood group antigens remains a fascinating subject with potential to contribute to the development of new interventions to reduce the global burden of malaria. PMID- 19812492 TI - Care of the pregnant thyroid cancer patient. AB - PURPOSE OF REVIEW: Thyroid cancer is the fastest rising type of cancer among women in North America. Care of the pregnant patient with thyroid cancer is, therefore, of concern to obstetricians, internists and endocrine specialists. Guidelines for the care of the pregnant patient with thyroid disease were released by the Endocrine Society in late 2007, and a symposium on thyroid dysfunction and pregnancy was hosted by the American Thyroid Association in April 2009. With this increasing interest in thyroid disease and pregnancy, a variety of important studies have been published recently. RECENT FINDINGS: In addition to guidelines published by the Endocrine Society, recent research has focused on detection and management of hypothyroidism in pregnancy, and consequences of hypothyroidism for the fetus and child. Impact of radioactive iodine therapy on subsequent fertility has been described. The risk of adverse outcomes due to thyroid surgery during pregnancy was evaluated in analysis of a large inpatient database. SUMMARY: Identification of hypothyroidism during pregnancy continues to be challenging due to the need for well established trimester-specific normal ranges. Physicians may reassure patients about the effects of radioactive iodine therapy on fertility, although men may wish to cryopreserve sperm prior to treatment. Thyroid surgery during pregnancy was associated with a two-fold increased risk of surgical complications. PMID- 19812493 TI - New insights in the management of elderly patients with acute myeloid leukemia. AB - PURPOSE OF REVIEW: The management of elderly patients with acute myeloid leukaemia (AML) is more than ever a challenge. Results associated with intensive chemotherapy remain disappointing, at least in nonselected patients. On the contrary, there are still no real standard alternatives even if some new approaches seem promising. RECENT FINDINGS: Recent studies aim to guide the selection of patients likely to benefit from intensive chemotherapy; to optimize remission induction and maintenance in those patients; and to evaluate long-term results associated with alternative therapies. Main results are the following: favorable AML subsets identified in younger patients remain favorable in elderly patients and might benefit from a relatively intensified treatment; if daunorubicin is used for induction, a dose higher than 45 mg/m2 x 3 results in a higher complete remission rate and longer survival, at least until 65 years of age; repeated intensive postremission courses do not seem to be effective in older patients; and median survival observed after clofarabine or azacitidine based therapy could be long enough to deserve prospective comparisons against conventional chemotherapy. SUMMARY: These new results may help to offer a personalized management to elderly patients with AML and to design future trials. PMID- 19812494 TI - Islet assessment for transplantation. AB - PURPOSE OF REVIEW: There is a critical need for meaningful viability and potency assays that characterize islet preparations for release prior to clinical islet cell transplantation. Development, testing, and validation of such assays have been the subject of intense investigation for the last decade. These efforts are reviewed, highlighting the most recent results while focusing on the most promising assays. RECENT FINDINGS: Assays based on membrane integrity do not reflect true viability when applied to either intact islets or dispersed islet cells. Assays requiring disaggregation of intact islets into individual cells for assessment introduce additional problems of cell damage and loss. Assays evaluating mitochondrial function, specifically mitochondrial membrane potential, bioenergetic status, and cellular oxygen consumption rate, especially when conducted with intact islets, appear most promising in evaluating their quality prior to islet cell transplantation. Prospective, quantitative assays based on measurements of oxygen consumption rate with intact islets have been developed, validated, and their results correlated with transplant outcomes in the diabetic nude mouse bioassay. CONCLUSION: More sensitive and reliable islet viability and potency tests have been recently developed and tested. Those evaluating mitochondrial function are most promising, correlate with transplant outcomes in mice, and are currently being evaluated in the clinical setting. PMID- 19812495 TI - Immunosuppression in high-risk transplantation. AB - PURPOSE OF REVIEW: Increasingly, transplant clinicians are faced with providing candidates with increased risks for poorer outcome with donor grafts that also carry higher risks of failure. Understanding the role of immunosuppressive management in these combinations of higher risks remains important for optimizing results. RECENT FINDINGS: Few immunosuppressive protocols have been rigorously tested in the high-risk renal transplant setting. The two main risk categories accounted for in the trials are those ones that confer increased risks to renal function, usually carried by the donor organs, and those protocols defined by increased risk for immunological failure, mostly determined by recipient characteristics. The studied protocols generally involve reduction or avoidance of nephrotoxic drugs in the first case and use of lymphocyte-depleting agents in the case of increased immunological risk. In both scenarios, acceptable short term results have been achieved. However, long-term results for high-risk transplants defined either by donor or recipient factors have yet to be reported. SUMMARY: The lack of long-term data for optimizing the right immunosuppressive regimen for a given donor/recipient risk profile remains an ongoing challenge for researchers and clinicians alike. PMID- 19812496 TI - Histoplasmosis in solid organ transplant recipients: early diagnosis and treatment. AB - PURPOSE OF REVIEW: To present current knowledge about the epidemiology, clinical presentation, diagnosis and treatment of histoplasmosis in solid organ transplant (SOT) recipients. RECENT FINDINGS: Histoplasmosis is rare in SOT patients, and most cases have been reported from large transplant centers in the Midwestern USA, where the fungus is endemic. Urine antigen testing and the chest computed tomography scan are emerging as especially useful diagnostic tools in the SOT population. Standard treatments include liposomal amphotericin b followed by itraconazole, but newer azoles (voriconazole and posaconazole) have good in-vitro activity and have been successfully used in some SOT cases. SUMMARY: Clinical suspicion is essential to early recognition of histoplasmosis in SOT patients who often present with fever of unknown cause and pulmonary symptoms. Diagnosis is usually made by a combinatorial approach, including antigen tests, radiology and appropriate biopsies for culture and histology. Treatment with available antifungals is associated with more than 95% success. PMID- 19812497 TI - Coccidioidomycosis in transplant recipients: a primer for clinicians in nonendemic areas. AB - PURPOSE OF REVIEW: Coccidioidomycosis is a fungal infection caused by Coccidioides spp., a fungus endemic to the southwestern USA. Immunocompromised patients with coccidioidomycosis often present with atypical manifestations. We provide clinical, diagnostic, and management approaches for clinicians in nonendemic areas who might encounter coccidioidomycosis in transplant recipients. RECENT FINDINGS: With preemptive screening, heightened awareness, and targeted prophylaxis, the incidence of coccidioidomycosis in transplant recipients has decreased to 1-3% in endemic areas. Dissemination and mortality continues to be clinically significant (about 30%). Serological findings are often unreliable in transplant recipients, necessitating invasive procedures for a tissue or microbiological diagnosis. Management recommendations are similar to those for immunocompetent hosts, except that transplant patients may require more prolonged therapy and secondary prophylaxis to prevent reactivation. The exact risk of donor-derived coccidioidomycosis is unknown. Routine antifungal prophylaxis is not currently recommended for transplant recipients visiting or relocating to endemic areas. SUMMARY: Transplant recipients who travel to or reside part-time or full-time in endemic areas are at risk for both primary and reactivated coccidioidomycosis. Clinicians in nonendemic areas should be aware that early diagnosis followed by prompt antifungal therapy could be lifesaving. Prophylactic strategies can prevent reactivation in select circumstances. PMID- 19812498 TI - An update on primary care management for tuberculosis in children. AB - PURPOSE OF REVIEW: Tuberculosis (TB) is still a significant cause of mortality and morbidity among children worldwide. Recent advances in scientific and clinical research have looked into the epidemiology, diagnostic, therapeutic and preventive interventions in order to provide insights into more effective control. RECENT FINDINGS: The risk of infection has been shown to be related to the duration and proximity of exposure to an infectious case, usually an adult or adolescent, rising with increased exposure in toddlers, associated with school entry, and in the teen years and early adulthood. Initial primary exposure to TB disease plays a significant role in the development of a latent TB infection, which may serve as a reservoir for future reactivation in adulthood. Diagnosis remains a dilemma for the clinician due to the lack of a gold standard. T-cell assays measuring production of interferon gamma have proven to be more specific than the Tuberculin Skin Test but are unable to distinguish between latent and active disease. SUMMARY: Recent findings have shown the importance of addressing not only active TB disease in children but also latent TB infection. Advances in understanding TB in children provide insights to enhance efforts to control this disease. PMID- 19812499 TI - Developmental defects and childhood cancer. AB - PURPOSE OF REVIEW: Childhood cancer syndromes are rare but easily recognizable in the clinical setting. Early detection of a potential underlying genetic defect by the general practitioner can be lifesaving. Additionally, important clinical clues that can aid in recognition, or lead to diagnostic referral, for a possible cancer-predisposing syndrome in the individual patient or family are discussed. RECENT FINDINGS: Recent advancements in genetics have increased our understanding of many genetic developmental pathways involved in cancer predisposition syndromes. Importantly, some of the defects in these pathways can explain the wide phenotypic variability in overall growth, physical dysmorphisms, and cancer risk in children. Genetic testing is now available for many disorders and important guidelines for screening are quickly evolving. The current diagnostic criteria, genetics, and cancer screening guidelines for neurofibromatosis type 1, Beckwith-Wiedemann syndrome/ hemihypertrophy, and PTEN hamartoma tumor syndrome, among others, are reviewed. SUMMARY: Early recognition of a possible underlying genetic defect with a resultant diagnostic work-up can lessen or prevent tumor burden, improve screening for possible cancer recurrence, and optimize care for children and their family members. Important clues from the history and physical include a family history of cancer, a specific cancer type frequently associated with a genetic defect, synchronous or metachronous cancers in the same individual, growth abnormalities, abnormal skin pigmentation, and/or thumb/radius malformations. Many cases can be recognized by the general practitioner and referred to the appropriate specialists for completion of the diagnostic work-up and recommendations for appropriate management. PMID- 19812500 TI - Ocular and cervical vestibular-evoked myogenic potentials: a study to determine whether air- or bone-conducted stimuli are optimal. AB - OBJECTIVES: The aim of this study was to determine whether bone-conducted vibration (BCV) or air-conducted sound (ACS) is the optimal mode for eliciting both ocular vestibular-evoked myogenic potentials (oVEMPs) and cervical VEMPs (cVEMPs). DESIGN: Twelve healthy volunteers, five patients with unilateral chronic otitis media, and 10 patients with unilateral Meniere disease underwent oVEMP and cVEMP tests using ACS and BCV stimulation modes in a random order. RESULTS: In healthy controls, BCV mode at Fz had a significantly higher response rate and larger nI-pI amplitude of oVEMPs than that of the ACS mode. In cVEMPs, a significantly higher response rate was noted in BCV mode at inion, when compared with ACS mode. However, no significant difference was noted in the p13-n23 amplitude between these two modes. In five chronic otitis media ears, absence of oVEMPs and cVEMPs in ACS mode and presence of oVEMPs and cVEMPs in BCV mode were shown. In 10 patients with Meniere disease, BCV mode elicited higher response rates of oVEMPs and cVEMPs in the pathological ears than ACS mode did. CONCLUSIONS: Using BCV mode, Fz and inion may be the optimal sites for eliciting oVEMPs and cVEMPs, respectively. Thus, BCV mode can be substituted for ACS mode to elicit oVEMPs and cVEMPs in the future, especially in "mass detection." In contrast, ACS mode provides an essential clinical merit of site selectivity. PMID- 19812501 TI - Severe hyperopic shift and irregular astigmatism after radial keratotomy. AB - OBJECTIVES: To report a case of acute high hyperopia occurring after radial keratotomy and evaluate the extent of corneal wavefront aberration with a Hartmann-Shack aberrometer. METHODS: A 45-year-old man had undergone bilateral radial keratotomy with four incisions for myopia 15 years earlier. The patient developed significant hyperopia and irregular corneal astigmatism in the left eye after the operation, reducing his best-corrected visual acuity. RESULTS: The best corrected visual acuity was 20/20 (with refraction of +0.25-1.50 diopters [D] x 169) in the right eye and 10/20 (with refraction of +9.5D-3.00D x 50) in the left eye. Topography showed marked flattening of the left cornea, and simulated retinal images of Landolt C circles were distorted in the left eye. CONCLUSIONS: This is a rare case in which radial keratotomy induced severe hyperopic shift. Wavefront analysis with a Hartmann-Shack aberrometer revealed severe irregular astigmatism with small clear zones. PMID- 19812502 TI - Association of renal damage with cardiovascular diseases is independent of individual cardiovascular risk profile in hypertension: data from the Italy - Developing Education and awareness on MicroAlbuminuria in patients with hypertensive Disease study. AB - OBJECTIVES: In the past years, several risk charts have been created to increase the accuracy of cardiovascular risk stratification. The most widely used and validated algorithms do not included target organ damage as risk prediction. The aim of the present study was to evaluate whether preclinical renal damage is associated with cardiovascular diseases independently of individual risk profile assessed by risk charts. METHODS: The study population was that of Italy Developing Education and awareness on MicroAlbuminuria in patients with hypertensive Disease, a large observational study conducted on hypertensive patients in Italy. The Framingham Risk Score (FRS), Systematic COronary Risk Estimation (SCORE) and Progetto Cuore Risk Score (Progetto Cuore RS) were computed in each eligible patient. Chronic kidney disease was defined by the presence of albuminuria or by a reduction of glomerular filtration rate. RESULTS: Study participants were categorized to have low, medium and high risk according to the tertiles of the three charts. Prevalence of total cardiovascular diseases progressively and significantly increased according to the degrees of risk assessed by the three charts, the highest prevalence being in participants with a high-risk profile (both high and medium vs. low risk <0.01 for FRS, SCORE and Progetto Cuore RS). The presence of chronic kidney disease was associated with total cardiovascular diseases, independently of FRS (odds ratio 1.64, 95% confidence interval 1.33-2.02, P < 0.001), SCORE (odds ratio 1.55, 95% confidence interval 1.21-1.98, P < 0.001) and Progetto Cuore RS (odds ratio 1.59, 95% confidence interval 1.22-2.07, P < 0.001). Moreover, inclusion of renal damage in the logistic model significantly increased the accuracy of the FRS (P < 0.05), SCORE (P < 0.01) and Progetto Cuore RS (P < 0.01) to identify patients with overt cardiovascular diseases. CONCLUSION: Identification of patients with preclinical renal damage should be encouraged in the hypertension cardiovascular risk stratification setting in order to achieve a more accurate individual risk computation. The presence of renal damage could improve cardiovascular risk prediction over the widely used risk stratification charts. PMID- 19812503 TI - Effect of pravastatin on nephroprotection in deoxycorticosterone acetate-salt hypertensive rats. AB - OBJECTIVE: Endothelin-1 (ET-1) has been implicated in the pathogenesis of renal impairment. The current study was undertaken to assess the effect of pravastatin on the progression of renal impairment in deoxycorticosterone acetate (DOCA)-salt hypertensive rats. METHODS: Four weeks after the start of DOCA-salt treatment and uninephrectomization, male Wistar rats were treated with one of the following therapies for 8 weeks: vehicle; a nonselective endothelin receptor antagonist bosentan; pravastatin; or hydralazine. RESULTS: Treatment with bosentan or pravastatin was associated with reductions in blood pressure and renal medullary hydroxyproline content, and improvement in glomerular filtration rate, urinary protein excretion, macrophage infiltration, tubular injury, and vascular injury, but not glomerulosclerosis. The renal medullary ET-1 protein levels and preproET 1 mRNA assessed by western blotting and real-time quantitative reverse transcription-PCR were significantly decreased (both P < 0.001) in the pravastatin-treated rats compared with vehicle, which was also confirmed by immunohistochemical analysis. However, there were no significant differences of ET-1 levels in the renal cortex among the DOCA-salt groups. The nephroprotective effects of pravastatin were not associated with its antihypertensive action because hydralazine despite reducing blood pressure failed to improve renal function and disorder. CONCLUSION: These results suggest a crucial role of renal endothelin system in the pathogenesis of renal functional and structural alterations in the DOCA-salt hypertensive rats. Pravastatin administration ameliorates the impairment of renal function and structures by attenuating medullary ET-1 expression, independent of systemic blood pressure. PMID- 19812504 TI - Pathway targets to explore in the treatment of small cell and large cell lung cancers. AB - INTRODUCTION: With low 5-year survival rates and little progress in therapy, improvements in therapeutic strategies for the treatment of small cell lung cancer (SCLC) and large cell lung cancer (LCC) are warranted. We hypothesized that different SCLC and LCC microarray data sets will share commonly perturbed pathways leading to the identification of new targets for therapeutic development. METHODS: Per gene mean expression fold change ratios were calculated from four publicly available microarray data sets consisting of a total of 113 profiled samples. This was followed by mapping of differentially expressed genes into functionally annotated biologic pathways using Ingenuity pathway analysis software. Common pathways containing genes whose expression levels differed between phenotypic classes defined by histology and gender were determined. RESULTS: Several significant common pathways overlapping these data sets were identified in silico. One of which, the Wnt/beta-catenin signaling pathway, is targeted by agents such as vorinostat and dasatinib, which are currently under exploration in SCLC trials. The remaining pathways are targeted by several drugs developed or under development in cancer therapeutics. CONCLUSION: This in silico pathway exploration in SCLC and LCC holds promise. Additional expression profiling of SCLC and LCC cases would likely add to the knowledge base. Further investigation into identified targets is warranted. PMID- 19812505 TI - Electrically assisted cycling: a new mode for meeting physical activity guidelines? AB - PURPOSE: The purpose of this study was to assess the potential of the electrically assisted bicycle (EAB) as a novel tool for meeting the physical activity guidelines in terms of intensity. METHODS: Twelve habitually active adult subjects were requested to cycle a track of 4.3 km at an intensity they would normally choose for commuter cycling, using three different support settings: no support (NO), eco support (ECO), and power support (POW). For estimating the intensity, the oxygen consumption was measured by using a portable gas-analyzing system, and HR was simultaneously measured. The bicycle was equipped with the SRM Training System to measure subjects' power output, pedaling rate, and the cycle velocity. RESULTS: Mean intensity was 6.1 MET for NO, 5.7 MET for ECO, and 5.2 MET for POW. Intensity was significantly lower in POW compared with that in NO. No differences were found between NO and ECO and between ECO and POW. Mean HR was significantly higher in NO compared with that in ECO and POW. The cycling speed with electrical support settings was significantly higher than cycling in the NO condition. Mean power output during cycling was significantly different among all three conditions. Most power outputs were supplied in the NO condition, and the lowest power output was supplied in the POW condition. CONCLUSIONS: Intensity during cycling on an EAB, in all three measured conditions, is sufficiently high to contribute to the physical activity guidelines for moderate-intensity health-enhancing physical activity for adults (cutoff, 3 MET). Further study is needed to conclude whether these results still hold when using the EAB in regular daily life and in subjects with other fitness level. PMID- 19812506 TI - Relation between individualized training impulses and performance in distance runners. AB - PURPOSE: The aim of this study was to develop a method to monitor responses to training loads on an individual basis in recreational long-distance runners (LDR) through training impulses (TRIMP) analysis. The hypothesis tested was that TRIMP on the basis of individually determined weighting factors could result in a better quantification of training responses and performance in LDR in comparison to methods on the basis of average-based group values. METHODS: The training load responses of eight LDR (aged 39.9 +/- 6.5 yr) were monitored using a modified version of the average-based TRIMP called individualized TRIMP (TRIMPi) during a period of 8 wk. The TRIMPi was determined in each LDR using individual HR and lactate profiles determined during an incremental treadmill test. Training induced effects on performance (5- and 10-km races) and changes in submaximal aerobic fitness (speeds at selected blood lactate concentrations of 2 and 4 mmol x L(-1)) were assessed before and at the end of the training intervention. RESULTS: Speed at 2 mmol x L(-1) (+21.3 +/- 5.2%, P < 0.001) and 4 mmol x L(-1) (+10.6 +/- 2.4%, P < 0.01) concentrations significantly increased after training. Improvements in running speed (%) at 2 mmol x L(-1) (r = 0.87, P = 0.005) and 4 mmol x L(-1) (r = 0.74, P = 0.04) concentrations were significantly related to weekly TRIMPi sum. No significant relationship between any variable was detected when average-based group values were used. The TRIMPi was significantly related to 5000- (r = -0.77; P = 0.02) and 10,000-m track performances (r = -0.82; P = 0.01). CONCLUSIONS: Individualized TRIMP is a valid tool in tracking fitness (speed at 2 and 4 mmol x L(-1)) and performance (i.e., 5000- and 10,000-m races) in LDR and is more valuable than the methods on the basis of average-based group values. TRIMPi could predict race performance in LDR. PMID- 19812507 TI - Seasonal variation of VO 2 max and the VO2-work rate relationship in elite Alpine skiers. AB - PURPOSE: Alpine ski performance relates closely to both anaerobic and aerobic capacities. During their competitive season, skiers greatly reduce endurance and weight training, and on-snow training becomes predominant. To typify this shift, we compared exhaustive ramp cycling and squat (SJ) and countermovement jumping (CMJ) performance in elite males before and after their competitive season. RESULTS: In postseason compared with preseason: 1) maximal oxygen uptake (VO 2 max) normalized to bodyweight was higher (55.2 +/- 5.2 vs 52.7 +/- 3.6 mL x kg( 1) x min(-1), P < 0.01), but corresponding work rate (W) was unchanged; 2) at ventilatory thresholds (VT), absolute and relative work rates were similar but heart rates were lower; 3) VO2/W slope was greater (9.59 +/- 0.6 vs 9.19 +/- 0.4 mL O2 x min(-1) x W(-1), P = 0.02), with similar flattening (P < 0.01) above V T1 at both time points; and 4) jump height was greater in SJ (47.4 +/- 4.4 vs 44.7 +/- 4.3 cm, P < 0.01) and CMJ (52.7 +/- 4.6 vs 50.4 +/- 5.0 cm, P < 0.01). DISCUSSION: We believe that aerobic capacity and leg power were constrained in preseason and that improvements primarily reflected an in-season recovery from a fatigued state, which was caused by incongruous preseason training. Residual adaptations to high-altitude exposure in preseason could have also affected the results. Nonetheless, modern alpine skiing seemingly provides an ample cardiovascular training stimulus for skiers to maintain their aerobic capacities during the racing season. CONCLUSIONS: We conclude that aerobic fitness and leg explosiveness can be maintained in-season but may be compromised by heavy or excessive preseason training. In addition, ramp test V O2/W slope analysis could be useful for monitoring both positive and negative responses to training. PMID- 19812508 TI - Effects of sit-up training versus core stabilization exercises on sit-up performance. AB - PURPOSE: Core stabilization exercises target abdominal and trunk muscles without the excessive loading that occurs during sit-ups. However, core stabilization exercise programs (CSEP) have not been widely adopted in the US Army partially because of the perceived deleterious impact they would have on performance during the Army Physical Fitness Test. The purpose was to determine whether performing CSEP in lieu of sit-ups during unit physical training would have detrimental effects on sit-up performance and passing rates on the fitness test. METHODS: Soldiers (N = 2616) between 18 and 35 yr of age were randomized to receive a traditional exercise program (TEP) with sit-ups or CSEP. Subjects with a previous history of low back pain or other injury precluding participation in training were excluded. The training programs were completed four times per week for 12 wk. Performance was assessed at baseline and after 12 wk. RESULTS: Both groups demonstrated significant improvements in sit-up performance and overall fitness scores over time (P < 0.001). There were no significant between-group differences in overall fitness scores (P = 0.142) or sit-up performance (P = 0.543). However, CSEP resulted in a significant improvement in sit-up passing rates by 5.6% compared with 3.9% for the TEP group (P = 0.004). CONCLUSIONS: CSEP did not have a detrimental impact on sit-up performance or overall fitness scores or pass rates. There was a small but significantly greater increase in sit-up pass rate in the CSEP (5.6%) versus the TEP group (3.9%). Incorporating CSEP into Army physical training does not increase the risk of suboptimal performance on the Army's fitness test and may offer a small benefit for improving sit-up performance. PMID- 19812509 TI - EMG-torque relationship and reliability of the medial and lateral hamstring muscles. AB - PURPOSE: The objectives of the present investigation were to examine the EMG joint torque relation and day-to-day reliability of the medial (MH) and lateral hamstring (LH) muscles during isometric contractions. METHODS: Twenty young adults performed five maximal voluntary isometric hamstring contractions (MVC) followed by contractions at 10%-90% MVC, in a random order. Full-wave-rectified and -integrated EMG during the middle 3 s of each contraction was expressed as a percent of the EMG during the three highest averaged MVC. A three-factor ANOVA (muscle x intensity x gender) with repeated measures was performed on normalized EMG activity. Reliability coefficients were determined for 10 randomly selected subjects that repeated the testing procedures 1 wk after the first session. RESULTS: The MH and LH EMG were observed to be significantly greater than equivalent percent MVC values at all intensity levels. A significant linear increase in MH and LH EMG was observed across contraction intensity levels, with no muscle or gender differences. Reliability was high for MH EMG across contraction intensities of 10%-60% MVC (intraclass correlation (ICC) = 0.70 0.82), moderate for the LH across 10%-30% MVC (ICC = 0.57-0.68) and the MH at 70% MVC (ICC = 0.52), and unreliable across all other contraction intensities for both muscles. CONCLUSIONS: The linear increase in MH and LH EMG, as a function of contraction intensity, was similar between both muscles but was highly reliable only for the MH during low to moderate intensities. PMID- 19812510 TI - A comparison of successful and unsuccessful attempts in maximal bench pressing. AB - PURPOSE: This study was designed to compare the differences in EMG and kinematics between successful and unsuccessful attempts in bench pressing at one repetition maximum (1RM) in recreational weight-trained subjects. We hypothesized that failure occurs during the sticking period (the period during which there is a temporary reduction in movement velocity). METHODS: Eleven male subjects (age = 21.9 +/- 1.8 yr, mass = 80.0 +/- 11.2 kg, height = 1.79 +/- 0.08 m) with at least 1 yr of bench press training experience participated in this study. They performed attempts at 1RM and 1RM + 2.5 kg in bench press during which kinematics and muscle activity were recorded. One successful attempt and one unsuccessful attempt were used for further analysis. RESULTS: Both attempts showed the same sticking period, but only half of the failures occurred during that period. The main differences in the kinematics occurred during the sticking period. Muscle activity, in contrast, showed the same pattern in both attempts and only differed during the downward and the start of the upward movement of the lift. CONCLUSIONS: The sticking period occurs in both successful and unsuccessful attempts in maximal bench press. However, failure does not always occur during the sticking period. PMID- 19812511 TI - Muscle forces or gravity: what predominates mechanical loading on bone? AB - Most mechanical forces acting on the skeleton are generated either through impact with the ground (i.e., gravitational loading) or through muscle contractions (i.e., muscle loading). If one of these conduits for activating mechanotransduction in bone is more effective than the other with respect to developing or maintaining bone strength, this would have important clinical implications for prescribing physical activity for the prevention or treatment of osteoporosis. This section of the symposium considered whether there is evidence from studies of humans that the effectiveness of physical activity to preserve bone health is dependent on whether the activities stimulate the skeleton primarily through gravitational or muscle loading. Conclusive evidence is lacking, but several lines of research suggest that physical activities that involve impact forces, and therefore generate both gravitation and muscle loading, are most likely to have beneficial effects on bone metabolism and reduce fracture risk. PMID- 19812512 TI - Is bone's response to mechanical signals dominated by muscle forces? AB - Skeletal loading in vertebrates controls modeling drifts, modulated remodeling rates, and affects growth trajectories. It is unclear whether the majority of the mechanical stimulus detected by bone cells originates from muscle contraction forces or from gravitational forces associated with substrate impact. Several clinical and basic science reports indicate that muscle forces play a dominant role in generating the mechanical stimulus in exercise-induced bone gain. Although it is, in most cases, difficult to separate the effects of gravitational forces acting on body mass from muscle contractions, several well-conceived experiments offer considerable insight into the propensity of muscle-derived forces per se to drive the adaptive response in bone. Load-induced osteogenesis requires that mechanical signals come packaged with particular characteristics, all of which can be generated from either gravitational or muscle forces. Neither of these two sources have been demonstrated empirically to be the source of bone's adaptive response, but a convincing body of data suggests that muscle contractions are present, significant, and capable of accounting for most of the adaptive responses. PMID- 19812513 TI - Is bone's response to mechanical signals dominated by gravitational loading? AB - During locomotion and exercise, bone is subjected to forces induced by gravitational loading and muscle loading. The inherent link between these modes of loading has confounded emergence of either one as the principal anabolic or anticatabolic signal in bone. A paradigm has emerged in the literature stipulating that muscle loading is the larger of the two, and therefore, bone morphology is predominantly determined by muscle loads. In spite of the intuitive appeal of a muscle-bone unit tuned to the magnitude of contractile forces, little evidence exists for the relatively few, large-magnitude muscle contractions arising during daily activities to dominate the mechanosensory input of bone. Moreover, a review of the literature raises several inconsistencies in this paradigm and indicates that the alternative--gravitational loading--can have a significant role in determining bone mass and morphology. Certainly, the relative contribution of each type of loading will depend on the specific activity, the location of the bone within the skeleton, and whether the bone is weight-bearing or not. Most likely, a more comprehensive paradigm for explaining sensitivity of bone to loading will have to include not only large-magnitude gravitational and muscle loads, but also other factors such as high-frequency, low-magnitude signals generated by the muscles during postural adjustments. PMID- 19812514 TI - Muscle forces or gravity--what predominates mechanical loading on bone? Introduction. AB - This article describes the background, rationale, significance, and objective of the recent American College of Sports Medicine symposium entitled "Muscle Forces or Gravity-What Predominates Mechanical Loading on Bone?" (55th Annual Meeting, Indianapolis, IN, May 28, 2008) and introduces a series of papers representing the positions taken by three of the speakers at that symposium. Our goal was to reinvigorate discussion on a topic that will inform many, provoke some, and, most importantly, stimulate ideas that will encourage progress in the field of exercise prescription for bone. PMID- 19812515 TI - Bone volumetric density, geometry, and strength in female and male collegiate runners. AB - PURPOSE: To explore differences in tibial bone geometry, volumetric density, and estimates of bone strength in runners and healthy controls. METHODS: Male (n = 21) and female (n = 38) runners (49.1 +/- 13.2 miles x wk(-1)) and inactive healthy controls (17 males and 32 females; mean age = 22 +/- 3.3 yr) were recruited to participate. Peripheral quantitative computed tomography was used to assess total volumetric bone mineral density (vBMD, mg x mm(-3)), total bone area (ToA, mm2), and an estimate of compressive bone strength (bone strength index (BSI) = ToA x total bone volumetric density (ToD2)) at the distal (4%) site of the tibia. ToA (mm2) and cortical bone area (CoA, mm2), cortical vBMD (CoD, mg x mm(-3)), cortical thickness (CoTh, mm), and an estimate of bone bending strength (polar strength strain index (SSIp), mm3) were measured at 50% and 66% sites. RESULTS: ToA and BSI were significantly greater (+11%-19%, P < 0.05) in female runners than controls at the 4% site. At the proximal sites, female runners had significantly greater ToA, CoA, CoTh, and SSIp (+9%-19%, all P < 0.001) compared with female controls. vBMD was similar at all tibia sites. Compared with controls, male runners had significantly greater CoTh at the 50% and 66% sites (+8% and 14%, respectively, P < 0.05) as well as greater CoA (+11%, P < 0.009) at the 66% site. There were no differences in bone strength or density at any site in the male runners. CONCLUSIONS: Greater bone strength in female runners was attributable to greater bone area rather than density. Although male runners did not show greater bone strength, they did exhibit favorable bone geometric properties. These data further document that running has osteogenic potential. PMID- 19812516 TI - Effect of preexercise electrolyte ingestion on fluid balance in men and women. AB - PURPOSE: This article aimed to study the effect of preexercise ingestion of an electrolyte-containing beverage and meal on fluid balance during exercise in men and women. METHODS: Twenty healthy, college-aged people (10 males, 10 females; mean +/- SD = 51.2 +/- 9.8 mL x kg x min(-1)) exercised at 58 +/- 4% V O 2 peak for 90 min, 45 min after ingesting 355 mL of chicken noodle soup (SOUP; 167 mmol x L(-1) Na +), carbohydrate-electrolyte beverage (CE; 16 mmol x L(-1) Na+), or water (WATER). After 90 min of exercise, participants completed a physical performance task (PPT) consisting of the calculated work that would be completed in 30 min at 60% V O 2 peak (n = 19). Water was allowed ad libitum throughout all trials. RESULTS: Fluid balance was improved in SOUP compared with WATER (-251 +/- 418 vs -657 +/- 593 g, respectively; P = 0.002) because of greater water intake and retention throughout the trial. Water intake was also greater in CE compared with WATER mostly because of an increase during the PPT. Plasma osmolality increased after ingestion of SOUP and remained elevated throughout exercise compared with both CE and WATER. Men and women had similar fluid balance results, with women having lower relative water intake and evaporative water losses compared with men. Physical performance was similar in all trials. CONCLUSIONS: SOUP ingested before exercise improves fluid balance because of increased ad libitum water intake and reduced proportional urinary water loss. The increase in water intake and, subsequently, the improved fluid balance may be because of a greater plasma osmolality before and throughout exercise. PMID- 19812517 TI - Restoration of strength despite low stress and abnormal imaging after Achilles injury. AB - PURPOSE: To determine the usefulness of clinical imaging in predicting the mechanical properties of rabbit Achilles tendons after acute injury. METHODS: We created a 2 x 7-mm full-thickness central tendon defect in one Achilles tendon of healthy rabbits. Rabbits in groups of 10 were killed immediately and 4 and 8 wk after surgery (n = 30). We then performed magnetic resonance (MR) imaging, ultrasonography (US), bone mineral densitometry (BMD), and mechanical testing to failure using a dual-cryofixation assembly on experimental and contralateral tendons. The main outcome measures included tendon dimensions, optical density (OD) of T1-weighted, proton density (PD), and T2-weighted MR sequences, US focal abnormalities, BMD of the calcaneus, and stress and peak load to failure. RESULTS: On MR imaging and US, all dimensions of the injured tendons after 2 wk and more were greater than those of the contralateral tendons (P < 0.05). The mean T1-weighted OD was greater at 4 wk (256 +/- 53) and 8 wk (184 +/- 24) than immediately after surgery (149 +/- 15). Mechanical stress was markedly lower in the experimental than in the contralateral tendons at both 4 wk (39 +/- 9 vs 77 +/- 16 N x mm(-2)) and 8 wk (58 +/- 6 vs 94 +/- 26 N x mm(-2); P < 0.05). Mean peak load to failure was significantly lower immediately after surgery (332 +/- 128 N) than at 4 and 8 wk (712 +/- 106 and 836 +/- 90 N, respectively). Both high T1-weighted OD (r = -0.73) and PD OD (r = -0.69) correlated with lower mechanical stress (P < 0.05). In the experimental tendons, higher T1-weighted OD correlated with lower peak load (r = -0.46; P < 0.05). CONCLUSIONS: Normal peak loads 4 wk after injury were withstood by an enlarged tendon of lower stress. These findings support progressive physical loading 4 wk after an Achilles tendon injury. T1 weighted OD constituted a marker of tendon mechanical recovery. PMID- 19812518 TI - Vascular remodeling in response to 12 wk of upper arm unilateral resistance training. AB - Participation in regular aerobic exercise has been shown to increase arterial size and that exercise-induced vascular remodeling may be regional rather than systemic. However, these issues have been minimally investigated concerning resistance training. PURPOSES: To determine whether 1) resistance training of the nondominant arm elicits an increase in diameter of the brachial artery and 2) unilateral training induces arterial remodeling in the contralateral arm. METHODS: Twenty-four previously untrained participants, consisting of 18 females (aged 22.3 +/- 5.1 yr) and 6 males (aged 21.7 +/- 1.8 yr), participated in unilateral strength training of the biceps and triceps for 12 wk using their nondominant arm. Isotonic (one-repetition maximum, 1RM) and isometric (ISO) strength of the biceps were assessed before and after training on both arms. Brachial artery diameter and biceps muscle cross-sectional area (CSA) of both arms were also measured before and after training using magnetic resonance imaging (MRI). RESULTS: Brachial artery diameter increased 5.47% (P < 0.05) in the nondominant trained arm with no change observed in the dominant untrained arm. Biceps CSA increased 18.3% (P < 0.05) in the trained arm with no change (P > 0.05) in the untrained limb. Nondominant 1RM and ISO strength increased by 35.1% and 16.8%, respectively (P < 0.05 for both), although there were no significant changes (P > 0.05) in the contralateral arm. A modest correlation was found between the increases in CSA and in brachial artery diameter (r2 = 0.19, P = 0.039). CONCLUSIONS: These results indicate that upper arm vascular remodeling, manifesting as increased brachial artery diameter, can result from resistance training and that these changes are localized to the trained limb and associated with increases in CSA. PMID- 19812519 TI - Muscular fitness in relation to physical activity and television viewing among young adults. AB - PURPOSE: To study how time spent in physical activity and that in television (TV) viewing are associated with muscular fitness among young adults. METHODS: The study population consisted of a cross-sectional sample of 381 males and 493 females aged 19.1 yr (SD 0.3) from the Northern Finland Birth Cohort 1986. Muscular fitness was measured by trunk muscle strength tests (trunk extension, flexion, and rotation) and jumping height test. Time spent on moderate- to vigorous-intensity physical activity and on TV viewing was self-reported by a questionnaire. RESULTS: The most physically active young adults performed significantly better in most trunk muscle strength tests and the jumping test than the least active subjects. The mean difference between the most and least active groups was at minimum 1.6 kg (95% confidence interval (CI) = -0.5 to 3.7) and at maximum 10.6 kg (95% CI = 4.7-16.5) for different trunk muscle strength tests and at minimum 4.4 cm (95% CI = 2.7-6.1) for the jumping height test. Males and females who watched TV for >or=2 h x d(-1) performed significantly worse in trunk extension and flexion tests and females also performed worse in the jumping test compared with those who watched TV <2 h x d(-1), independent of their physical activity level. The mean difference between low and high TV users was at minimum -3.8 kg (95% CI = -6.7 to -0.9) for trunk extension and flexion strength and -1.2 cm (95% CI = -2.0 to -0.4) for jumping height in females. CONCLUSIONS: Among young adults, daily TV viewing for >or=2 h, irrespective of physical activity level, was associated with poorer muscular fitness. PMID- 19812520 TI - Crural fascia and muscle origins related to medial tibial stress syndrome symptom location. AB - PURPOSE: Traction-induced injury, related to muscles of the superficial and deep posterior compartments, has been implicated as the cause of medial tibial stress syndrome (MTSS) with symptoms commonly occurring in the distal third of the posteromedial tibia. Standard anatomic texts do not identify this region as an attachment site for these structures. Research into the anatomical arrangement of these structures has been inconclusive. The deep crural fascia (DCF) has been implicated as a cause of traction-induced injury in MTSS but not fully researched. The purpose of this study was to define the tibial origins of the DCF and the muscles of the superficial and deep posterior compartments relative to MTSS-related pain commonly reported along the distal one half to one third of the diaphysis of the medial tibial border and to identify the prevalence of a soleal aponeurosis. METHODS: The tibial attachments of the DCF, the soleus, the flexor digitorum longus, and the tibialis posterior were quantified relative to the medial malleolus in sixteen cadaver specimens. RESULTS: Mean distal attachments to the medial tibial border were superior to the distal third of the tibia for the muscles of the posterior compartments, suggesting that the role of the soleus, the tibialis posterior, and the flexor digitorum longus in producing pain typically associated with MTSS may be limited. The DCF of all but three specimens attached along the entire length of the medial tibia investing the medial malleolus. CONCLUSION: Traction-induced injury theories involving the muscles of the superficial and deep posterior compartments are not supported by anatomical evidence in the present study. The tibial attachments of the DCF in this study support theories implicating DCF involvement in creating traction-induced injury. PMID- 19812521 TI - Efficacy of field treatments to reduce body core temperature in hyperthermic subjects. AB - PURPOSE: To contrast the effects of three postcooling techniques in reducing body core temperature (Tc) in exercise-induced hyperthermic participants on the cessation of exercise. METHODS: Eleven healthy active male volunteers were cooled during a 40-min period using three different methods: ice packs to the neck, axillae, and groin (ICE); water spray and fan (FAN); and 2 L of chilled (20 degrees C) intravenous saline administered during a 20-min period (IV). Rate of decrease in Tc, cardiovascular responses, and any incidence of reported adverse effects were investigated. Trials were presented in a counterbalanced order with the volunteers' body core temperature being elevated to 40.0 degrees C on three occasions via an intermittent walk-run (2 min at 6 km x h and 4 min at 10 km x h) protocol conducted within a climate-controlled chamber (34.2 +/- 0.5 degrees C and 62.3 +/- 3.1% relative humidity). RESULTS: Rate of Tc reduction during the first 20 min of cooling was greater for FAN compared with ICE (0.09 +/- 0.02 degrees C.min vs 0.07 +/- 0.02 degrees C.min, P < 0.05), whereas IV did not differ with the other trials (0.08 +/- 0.01 degrees C.min, P > 0.05). Three participants complained of numbness or paresthesia in their arm or hand during administration of the chilled saline, although these symptoms resolved within 5 min of ceasing the infusion. CONCLUSIONS: All three cooling techniques reduced Tc and would be suitable for first aid application in a field setting during transportation to adequate medical facilities. Chilled IV saline did not produce any contraindications, providing a suitable alternative for Tc cooling. PMID- 19812522 TI - Which is the optimal exercise to strengthen supraspinatus? AB - PURPOSE: Supraspinatus strengthening is an important part of shoulder rehabilitation programs. Because of the force-couple relationship between supraspinatus and deltoid during shoulder abduction, it has been proposed that the optimal exercise to strengthen supraspinatus is one that would maximize supraspinatus activity while minimizing surrounding muscle activity, particularly deltoid. The aim of this study was to simultaneously examine all exercises known to recruit supraspinatus at high levels and to compare the level of recruitment in supraspinatus, deltoid, and infraspinatus. METHODS: Using a combination of surface and intramuscular electrodes, EMG recordings were taken from supraspinatus, infraspinatus, and the three parts of deltoid of 15 subjects with normal dominant shoulder function during maximal isometric contractions in "empty can," "full can," prone elevation, pendant external rotation, and prone external rotation exercise positions. RESULTS: There was no significant difference in the level of supraspinatus activation for all exercise positions examined. No significant difference was found in activation levels across all muscles during the "can" exercise positions. The exercise positions using external rotation were found to activate parts of deltoid significantly less than the "can" and prone elevation positions. Infraspinatus was activated at greater levels than supraspinatus during the external rotation exercises. CONCLUSIONS: Using the criteria that the optimal exercise to strengthen supraspinatus should maximize supraspinatus activity while particularly minimizing deltoid activity, this study has shown that the pendant external rotation and prone external rotation exercises are more valid than the "can" and prone elevation exercises for supraspinatus strengthening in subjects with normal dominant shoulder function. PMID- 19812523 TI - CYP2A13, ADH1B, and ADH1C gene polymorphisms and pancreatic cancer risk. AB - OBJECTIVES: Pancreatic carcinoma etiology and molecular pathogenesis are weakly understood. Based on the assumption that genetic variation in carcinogen metabolism further modifies the risk of exposure-related cancers, we studied the association of polymorphisms in the tobacco carcinogen-metabolizing gene CYP2A13 (Arg101Stop) and the alcohol-metabolizing genes ADH1B (Arg48His) and ADH1C (Ile350Val) with pancreatic cancer risk. METHODS: Polymorphisms were studied by allelic discrimination. RESULTS: In a hospital-based case-control study, CYP2A13 variant alleles coding an inactive enzyme were found in 7 of 265 cancer-free controls and in none of 235 pancreatic carcinoma patients. Neither ADH1B or ADH1C polymorphisms alone nor their combinations showed a significant effect on pancreatic cancer risk. CONCLUSIONS: The first study of the roles of CYP2A13, ADH1B, and ADH1C in pancreatic cancer etiology suggested that the controls may have a lower ability to bioactivate tobacco-derived procarcinogens than the cases. PMID- 19812524 TI - Quantification of insulin gene expression during development of pancreatic islet cells. AB - OBJECTIVE: Despite great progress in understanding the transcriptional regulation of the development of insulin-secreting beta cells, the quantitative temporal expression of insulin gene(s) remains largely unknown. We here aimed to quantify insulin gene transcripts during development. METHODS: We described bioinformatics algorithms to quantify (insulin) gene transcript abundance in sequential microarray data sets at the global level. Several molecular techniques were used to confirm our analyses. RESULTS: We demonstrated that the expression of insulin genes was up-regulated at approximately 14-fold, 700- to 2000-fold, and 5000- to 6000-fold in Pdx1- and Ngn3-expressing cells and adult islets compared with definitive endodermal or embryonic stem cells, respectively. The expression of multiple genes encoding molecules involved in posttranslational modifications of insulin and glucose sensing was also elevated in the same period. All islet and associated genes determined with microarray data were confirmed not only to be up regulated by real-time quantitative reverse transcriptase polymerase chain reaction but also that the magnitude of their increase quantified with these 2 methods was statistically highly correlated. Consistent with the above, green fluorescence protein expression under the control of the mouse insulin 1 promoter could be visualized in the pancreas from embryonic day (E) 11.5, increasing progressively through E13.5 to E15.5. CONCLUSION: Our study provides a novel insight into islet developmental biology. PMID- 19812525 TI - Independent contribution of common CFTR variants to chronic pancreatitis. AB - OBJECTIVE: We have assessed whether CFTR gene has a major impact on chronic pancreatitis (CP) pathogenesis than that provided by the CFTR mutations. For this aim, we have evaluated clinical parameters, CFTR mutations, and 3 potential regulatory CFTR variants (coding single-nucleotide polymorphisms): c.1540A>G, c.2694T>G, and c.4521G>A. METHODS: CFTR gene analysis was performed in a cohort of 136 CP patients and 93 controls from Spanish population using current scanning techniques (single-strand conformation polymorphism/heteroduplex, denaturing gradient gel electrophoresis, and denaturing high-performance liquid chromatography) and direct sequencing. RESULTS: A higher frequency of CFTR mutations were observed in patients (39%) than in controls (15%; P < or = 0.001), differences being mostly attributable to the prevalence of the cystic fibrosis (CF)-causing mutations (P = 0.009). The analysis of variants has shown statistically significant differences between patients and controls for c.4521G>A (Pcorrected = 0.036). Furthermore, the multi-marker analysis revealed that the 1540A;2694G;4521A (AGA) haplotype was more prevalent in CP than controls (Pcorrected = 0.042). Remarkably, this association was unrelated to CF-causing mutations (P = 0.006). CONCLUSIONS: Our results corroborate the higher susceptibility of CF carriers to CP and, furthermore, suggest that the AGA haplotype could contribute to an increased risk in the development of CP irrespective of other CF-causing mutations. PMID- 19812526 TI - Analysis of humoral immune response in experimental autoimmune pancreatitis in mice. AB - OBJECTIVES: To study the autoimmune response in MRL/Mp mice, which spontaneously develop pancreatitis in the exocrine pancreatic tissue. METHODS: Six-week-old female mice were injected intraperitoneally with polyinosinic polycytidylic acid at a dose of 5 mg/kg of body weight twice a week for up to 12 weeks. The mice were serially killed, and the severity of their pancreatitis was graded with a histological scoring system. Immunohistological examinations were performed, and the serum levels of autoantibodies were measured by enzyme-linked immunosorbent assay. RESULTS: The administration of polyinosinic polycytidylic acid accelerated the development of pancreatitis, with abundant infiltration of B220 B cells and CD138 plasmacytes. Various autoantibodies directed against autoantigens, including carbonic anhydrase II and lactoferrin, were detected but none against glutamic acid decarboxylase. Of these, autoantibodies directed against the pancreatic secretory trypsin inhibitor (PSTI; 91.7%) were more prevalent than those against carbonic anhydrase II (33.3%) or lactoferrin (45.8%). Determination of the epitope of the anti-PSTI antibody showed that most immunoreactivity was directed at the site on PSTI that is active in the suppression of trypsin activity. CONCLUSIONS: The autoimmune response to PSTI protein may induce a failure of PSTI activity, resulting in the activation of trypsinogen and the subsequent disease progression. PMID- 19812534 TI - Antibody profiling by Luciferase Immunoprecipitation Systems (LIPS). AB - Technologies for comprehensively understanding and quantifying antibody profiles to autoantigens and infectious agents may yield new insights into disease mechanisms and may elucidate new markers to substratify disease with different clinical features and better understand pathogenesis. We have developed a highly quantitative method called Luciferase Immunoprecipitation Systems (LIPS) for profiling patient sera antibody responses to autoantigens and pathogen antigens associated with infection. Unlike ELISAs, the highly sensitive LIPS is easily implemented to survey humoral serological response profiles to different antigens in a universal format and produces dynamic antibody titer ranges up to 6 log(10) for some antigens. In these studies, quantitative profiling by LIPS of patient humoral responses against panels of antigens or even the entire proteome of some pathogens (i.e. HIV), is typically more informative than testing a single antigen by ELISA. In addition, LIPS also eliminates time and effort needed to produce highly purified antigens as well as the labor-intensive assay optimization steps needed for standard ELISAs. Here we provide a detailed protocol describing the technical aspects of performing LIPS assays for readily profiling antibody responses to single or multiple antigens. PMID- 19812535 TI - Assessing acid-base disorders. AB - Effective management of acid-base disorders depends on accurate diagnosis. Three distinct approaches are currently used in assessing acid-base disorders: the physiological approach, the base-excess approach, and the physicochemical approach. There are considerable differences among the three approaches. In this review, we first describe the conceptual framework of each approach, and comment on its attributes and drawbacks. We then highlight the application of each approach to patient care. We conclude with a brief synthesis and our recommendations for choosing an approach. PMID- 19812536 TI - New molecular players facilitating Mg(2+) reabsorption in the distal convoluted tubule. AB - The renal distal convoluted tubule (DCT) has an essential role in maintaining systemic magnesium (Mg(2+)) concentration. The DCT is the final determinant of plasma Mg(2+) levels, as the more distal nephron segments are largely impermeable to Mg(2+). In the past decade, positional candidate strategies in families with inherited forms of hypomagnesemia have led to the identification of genes involved in Mg(2+) handling. A large fraction of this resides in the DCT, namely, (i) the transient receptor potential channel melastatin subtype 6 (TRPM6), a divalent cation-permeable channel located at the luminal membrane of the DCT, facilitates Mg(2+) entry from the pro-urine into the cell; (ii) the epidermal growth factor is a novel hormone regulating active Mg(2+) transport through TRPM6; (iii) the voltage-gated K(+) channel, Kv1.1, establishes a favorable luminal membrane potential for TRPM6-mediated Mg(2+) transport; (iv) the Na(+)/K(+)-ATPase gamma-subunit (gamma-Na(+)/K(+)-ATPase) was identified as mutated protein in a family with isolated dominant hypomagnesemia. The molecular mechanism by which gamma-Na(+)/K(+)-ATPase is involved in DCT Mg(2+) handling remains unknown; (v) a high percentage of patients with mutations in the renal transcription factor HNF1B (hepatocyte nuclear factor 1 homeobox B) gene develop hypomagnesemia; and (vi) Gitelman and EAST/SeSAME syndrome patients suffer from a similar tubulopathy due to mutations in NCC (NaCl cotransporter) and Kir4.1, respectively. In these patients, decreased expression of TRPM6 is proposed to cause hypomagnesemia. Insights into the molecular mechanisms of the identified genes, as well as the identification of novel genes, will further improve our knowledge about renal Mg(2+) handling. PMID- 19812537 TI - Parathyroid hormone measurement in CKD. AB - The Kidney Disease Outcomes Quality Initiative (K/DOQI) guidelines recommend that serum parathyroid hormone (PTH) concentration of patients with chronic kidney disease (CKD) should be measured regularly and maintained within target ranges that are defined according to the stage of CKD (e.g., 150-300 pg/ml in patients with CKD stage 5). The quality of the PTH assay is of paramount importance, as it contributes to the therapeutic decision. Indeed, when the PTH concentration is above these target values, drugs that decrease PTH secretion, such as active vitamin D compounds or calcimimetic agents, may be given and the doses are then adapted according to the evolution of the PTH concentration. By contrast, if the PTH concentration is below the target range, any treatment that may decrease PTH secretion is stopped to avoid adynamic bone disease and associated extra-skeletal calcifications. The aim of this article is to discuss the main features and pitfalls related to PTH measurement in the setting of CKD. PMID- 19812538 TI - Endoplasmic reticulum stress in proteinuric kidney disease. AB - Endoplasmic reticulum (ER) stress refers to physiological or pathological states that result in accumulation of misfolded proteins in the ER. To handle misfolded proteins, the ER has in place quality control mechanisms, including the unfolded protein response and ER-associated degradation (ERAD). ER stress in renal pathophysiology is a relatively new area of research. Mice heterozygous for a mutation in the ER chaperone, BiP, develop glomerulosclerosis and tubulointerstitial disease. Induction of ER stress in glomerular cells has been described in experimental models of membranous nephropathy and membranoproliferative glomerulonephritis, and exogenous induction of ER stress ('preconditioning') reduced proteinuria. In human kidney biopsies, markers of ER stress in glomeruli have been identified in various noninflammatory and inflammatory glomerulopathies. A tubulointerstitial ER stress response, in some cases associated with tubular cell apoptosis, may occur in glomerular diseases associated with proteinuria, including puromycin aminonucleoside nephrosis, protein overload, and experimental and human diabetic nephropathy. Certain missense mutations in nephrin and podocin, as well as underglycosylation of nephrin, result in misfolding and retention in the ER, and eventually ERAD. Understanding the various aspects of ER stress will provide an opportunity for development of novel therapeutic strategies for proteinuric diseases. PMID- 19812539 TI - SAPS 3 scores at the start of renal replacement therapy predict mortality in critically ill patients with acute kidney injury. AB - Patients can experience acute kidney injury and require renal replacement therapy at any time during their admission to intensive care units. Prognostic scores have been used to characterize and stratify patients by the severity of acute disease, but scores based on findings during the day of admission may not be reliable surrogate markers of the severity of acute illness in this population. The aim of this study was to evaluate the performance of SAPS 3 and MPM(0)-III scores, determined at the start of renal replacement therapy, in 244 patients admitted to 11 units of three hospitals in Rio de Janeiro, Brazil. Continuous renal replacement therapy was used as first indication in 84% of these patients. Discrimination by area under the receiver operating characteristic curve was significantly better for SAPS 3 than for MPM(0)-III, as was the calibration measured by the Hosmer-Lemeshow goodness-of-fit test. Mortality prediction and calibration approached those eventually found when a customized equation of SAPS 3 for Central and South America was used. After adjusting for other relevant covariates in multivariate analyses, both higher prognostic scores and length of stay in the unit prior to the start of renal replacement therapy were the main predictive factors for hospital mortality. Our study shows that a customized SAPS 3 model was accurate in predicting mortality and seems a promising algorithm to characterize and stratify patients in clinical studies. PMID- 19812540 TI - Protein microarrays identify antibodies to protein kinase Czeta that are associated with a greater risk of allograft loss in pediatric renal transplant recipients. AB - Antibodies to human leukocyte antigens (HLAs) are a risk factor for acute renal allograft rejection and loss. The role of non-HLAs and their significance to allograft rejection have gained recent attention. Here, we applied protein microarray technology, with the capacity to simultaneously identify 5056 potential antigen targets, to assess non-HLA antibody formation in 15 pediatric renal transplant recipients during allograft rejection. Comparison of the pre- and post-transplant serum identified de novo antibodies to 229 non-HLA targets, 36 of which were present in multiple patients at allograft rejection. On the basis of its reactivity, protein kinase Czeta (PKCzeta) was selected for confirmatory testing and clinical study. Immunohistochemical analysis found PKCzeta both within the renal tissue and infiltrating lymphocytes at rejection. Patients who had an elevated anti-PKCzeta titer developed rejection, which was significantly more likely to result in graft loss. The absence of C4d deposition in patients with high anti-PKCzeta titers suggests that it is a marker of severe allograft injury rather than itself being pathogenic. Presumably, critical renal injury and inflammation associated with this rejection subtype lead to the immunological exposure of PKCzeta with resultant antibody formation. Prospective assessment of serum anti-PKCzeta levels at allograft rejection will be needed to confirm these results. PMID- 19812541 TI - Nephrin mutations cause childhood- and adult-onset focal segmental glomerulosclerosis. AB - Mutations in the NPHS1 gene cause congenital nephrotic syndrome of the Finnish type presenting before the first 3 months of life. Recently, NPHS1 mutations have also been identified in childhood-onset steroid-resistant nephrotic syndrome and milder courses of disease, but their role in adults with focal segmental glomerulosclerosis remains unknown. Here we developed an in silico scoring matrix to evaluate the pathogenicity of amino-acid substitutions using the biophysical and biochemical difference between wild-type and mutant amino acid, the evolutionary conservation of the amino-acid residue in orthologs, and defined domains, with the addition of contextual information. Mutation analysis was performed in 97 patients from 89 unrelated families, of which 52 presented with steroid-resistant nephrotic syndrome after 18 years of age. Compound heterozygous or homozygous NPHS1 mutations were identified in five familial and seven sporadic cases, including one patient 27 years old at onset of the disease. Substitutions were classified as 'severe' or 'mild' using this in silico approach. Our results suggest an earlier onset of the disease in patients with two 'severe' mutations compared to patients with at least one 'mild' mutation. The finding of mutations in a patient with adult-onset focal segmental glomerulosclerosis indicates that NPHS1 analysis could be considered in patients with later onset of the disease. PMID- 19812542 TI - COMP-angiopoietin-1 decreases lipopolysaccharide-induced acute kidney injury. AB - During sepsis endothelial dysfunction is an important pathogenetic mechanism in acute kidney injury (AKI). Lipopolysaccharide (LPS)-induced endotoxemia is associated with renal hemodynamic changes such as alterations of renal blood flow (RBF), vascular resistance, and glomerular filtration rate. We used adenoviral delivery of an engineered variant of native angiopoietin-1 (COMP-angiopoietin-1) containing anti-inflammatory and anti-permeability functions, to determine if regulation of renal endothelial cell dysfunction may have a beneficial role in preventing AKI during LPS-induced endotoxemia in mice. This treatment prevented the endotoxin-induced decrease of RBF and mean arterial pressure while improving glomerular filtration rate. Treatment also mitigated the effects of LPS on renal intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 protein expression, the number of ER-HR3-positive macrophages that infiltrated the kidney, serum nitrate/nitrite levels, renal inducible nitric oxide synthase protein expression, the induction of tubular epithelial reactive oxygen and nitrogen species, and renal microvascular permeability. Our findings show that COMP-angiopoietin-1, an endothelium-oriented therapeutic agent, protects against AKI caused by endotoxemia. PMID- 19812543 TI - The reward circuit: linking primate anatomy and human imaging. AB - Although cells in many brain regions respond to reward, the cortical-basal ganglia circuit is at the heart of the reward system. The key structures in this network are the anterior cingulate cortex, the orbital prefrontal cortex, the ventral striatum, the ventral pallidum, and the midbrain dopamine neurons. In addition, other structures, including the dorsal prefrontal cortex, amygdala, hippocampus, thalamus, and lateral habenular nucleus, and specific brainstem structures such as the pedunculopontine nucleus, and the raphe nucleus, are key components in regulating the reward circuit. Connectivity between these areas forms a complex neural network that mediates different aspects of reward processing. Advances in neuroimaging techniques allow better spatial and temporal resolution. These studies now demonstrate that human functional and structural imaging results map increasingly close to primate anatomy. PMID- 19812544 TI - A novel role for constitutively expressed epithelial-derived chemokines as antibacterial peptides in the intestinal mucosa. AB - Intestinal-derived chemokines have a central role in orchestrating immune cell influx into the normal and inflamed intestine. Here, we identify the chemokine CCL6 as one of the most abundant chemokines constitutively expressed by both murine small intestinal and colonic epithelial cells. CCL6 protein localized to crypt epithelial cells, was detected in the gut lumen and reached high concentrations at the mucosal surface. Its expression was further enhanced in the small intestine following in vivo administration of LPS or after stimulation of the small intestinal epithelial cell line, mIC(c12), with IFNgamma, IL-4 or TNFalpha. Recombinant- and intestinal-derived CCL6 bound to a subset of the intestinal microflora and displayed antibacterial activity. Finally, the human homologs to CCL6, CCL14 and CCL15 were also constitutively expressed at high levels in human intestinal epithelium, were further enhanced in inflammatory bowel disease and displayed similar antibacterial activity. These findings identify a novel role for constitutively expressed, epithelial-derived chemokines as antimicrobial peptides in the intestinal mucosa. PMID- 19812545 TI - Origins and functional impact of copy number variation in the human genome. AB - Structural variations of DNA greater than 1 kilobase in size account for most bases that vary among human genomes, but are still relatively under-ascertained. Here we use tiling oligonucleotide microarrays, comprising 42 million probes, to generate a comprehensive map of 11,700 copy number variations (CNVs) greater than 443 base pairs, of which most (8,599) have been validated independently. For 4,978 of these CNVs, we generated reference genotypes from 450 individuals of European, African or East Asian ancestry. The predominant mutational mechanisms differ among CNV size classes. Retrotransposition has duplicated and inserted some coding and non-coding DNA segments randomly around the genome. Furthermore, by correlation with known trait-associated single nucleotide polymorphisms (SNPs), we identified 30 loci with CNVs that are candidates for influencing disease susceptibility. Despite this, having assessed the completeness of our map and the patterns of linkage disequilibrium between CNVs and SNPs, we conclude that, for complex traits, the heritability void left by genome-wide association studies will not be accounted for by common CNVs. PMID- 19812546 TI - Saturn's largest ring. AB - Most planetary rings in the Solar System lie within a few radii of their host body, because at these distances gravitational accelerations inhibit satellite formation. The best known exceptions are Jupiter's gossamer rings and Saturn's E ring, broad sheets of dust that extend outward until they fade from view at five to ten planetary radii. Source satellites continuously supply the dust, which is subsequently lost in collisions or by radial transport. Here we report that Saturn has an enormous ring associated with its outer moon Phoebe, extending from at least 128R(S) to 207R(S) (Saturn's radius R(S) is 60,330 km). The ring's vertical thickness of 40R(S) matches the range of vertical motion of Phoebe along its orbit. Dynamical considerations argue that these ring particles span the Saturnian system from the main rings to the edges of interplanetary space. The ring's normal optical depth of approximately 2 x 10(-8) is comparable to that of Jupiter's faintest gossamer ring, although its particle number density is several hundred times smaller. Repeated impacts on Phoebe, from both interplanetary and circumplanetary particle populations, probably keep the ring populated with material. Ring particles smaller than centimetres in size slowly migrate inward and many of them ultimately strike the dark leading face of Iapetus. PMID- 19812548 TI - The twain shall meet. PMID- 19812547 TI - A regulatory circuit for piwi by the large Maf gene traffic jam in Drosophila. AB - PIWI-interacting RNAs (piRNAs) silence retrotransposons in Drosophila germ lines by associating with the PIWI proteins Argonaute 3 (AGO3), Aubergine (Aub) and Piwi. piRNAs in Drosophila are produced from intergenic repetitive genes and piRNA clusters by two systems: the primary processing pathway and the amplification loop. The amplification loop occurs in a Dicer-independent, PIWI Slicer-dependent manner. However, primary piRNA processing remains elusive. Here we analysed piRNA processing in a Drosophila ovarian somatic cell line where Piwi, but not Aub or AGO3, is expressed; thus, only the primary piRNAs exist. In addition to flamenco, a Piwi-specific piRNA cluster, traffic jam (tj), a large Maf gene, was determined as a new piRNA cluster. piRNAs arising from tj correspond to the untranslated regions of tj messenger RNA and are sense oriented. piRNA loading on to Piwi may occur in the cytoplasm. zucchini, a gene encoding a putative cytoplasmic nuclease, is required for tj-derived piRNA production. In tj and piwi mutant ovaries, somatic cells fail to intermingle with germ cells and Fasciclin III is overexpressed. Loss of tj abolishes Piwi expression in gonadal somatic cells. Thus, in gonadal somatic cells, tj gives rise simultaneously to two different molecules: the TJ protein, which activates Piwi expression, and piRNAs, which define the Piwi targets for silencing. PMID- 19812549 TI - Large trial to examine parasites' influence on global killers. PMID- 19812551 TI - Survey of medical centers points to funding gaps. PMID- 19812553 TI - Value of health interventions underestimated by governments. PMID- 19812554 TI - 'Propaganda index' proposed for medical literature. PMID- 19812555 TI - Harvard Medical School rescinds controversial media rules. PMID- 19812556 TI - Analysis of retractions puts spotlight on academia. PMID- 19812558 TI - Straight talk with...Christopher Murray. AB - Hard numbers can be difficult to come by in the current debate about health care in the US. Even rarer are accurate assessments of health care systems in less developed countries. But policy makers are not completely groping in the dark when it comes to data-thanks in part to Christopher Murray. Two years ago, Murray, a physician and health economist with experience at the World Health Organization (WHO), took the helm of the newly created Institute for Health Metrics and Evaluation at the University of Washington. Since 2007, the institute, funded largely by the Bill & Melinda Gates Foundation and the state of Washington, has grown to a staff of 75 people and has begun churning out studies that that are shaping the debate on health care reform. For instance, Murray's group-along with colleagues at his former base, Harvard University in Cambridge, Massachusetts-have documented huge disparities in life expectancy and mortality in parts of the US. In some pockets of the country, life expectancy for women is even on the decline (PLoS Med. 27, e66; 2008). Murray spoke with Charlotte Schubert about how having accurate numbers can add up to progress in health care. PMID- 19812559 TI - The most transparent research. AB - Biomedicine would be a breeze if organisms were transparent. With the ability to see through tissues, scientists could spot the development of tumors more easily in study animals. And biologists could study exactly how an animal's organs develop by observing them as they grow. In effect, the secrets of the body would be out there for everyone to see.The thought of peering into our tissues may sound like science fiction, but one day it could be science. Using ideas from genetics, electrical engineering, chemistry and solid-state physics, a handful of researchers are working on ways to render biological tissues transparent.Some have already succeeded: in 2007, Richard White, a biologist at the Dana Farber Cancer Institute in Boston, used careful breeding techniques to create a transparent adult zebrafish named casper, evoking a reference to the famous cartoon ghost by the same name. Now, more than 100 labs around the world are using these transparent fish to study cancer pathology and development in real time. "The field of in vivo imaging-looking at things that are happening inside an actual organism-is growing rapidly," White says.Researchers are even making strides toward turning human tissue transparent. The primary reason we can't see what's inside of us is that light scatters when it passes through tissue. The body is densely packed with many types of substances, such as bone and fat, and light travels through them at different speeds because they have what physicists refer to as different refractive indices. The result is that light can't pass through biological tissues in a straight line, much as car headlights don't pass through dense fog. To fix this problem, scientists are working on developing ways to stop tissues from scattering light. Indeed, "if you take away the scattering properties of human tissues, we would look more or less like jellyfish," explains Changhuei Yang, an electrical engineer and bioengineer at the California Institute of Technology.Though their approaches (described in the following pages) are diverse, these researchers share the common goal of making it one day possible to see what's going on deep inside of the body-a feat that would provide new insights into our biology and help doctors diagnose and treat disease much more easily. "We're trying to push the limits in terms of what can we uncover," says Bernard Choi, a bioengineer at the University of California-Irvine. PMID- 19812560 TI - CCL3L1 and HIV/AIDS susceptibility. PMID- 19812561 TI - CCL3L1 and HIV/AIDS susceptibility. PMID- 19812562 TI - Experimental aspects of copy number variant assays at CCL3L1. PMID- 19812563 TI - Reply to: "Experimental aspects of copy number variant assays at CCL3L1". PMID- 19812565 TI - Gateway to the diseased brain. PMID- 19812566 TI - Preserving fertility during cancer treatment. PMID- 19812567 TI - Epilepsy: synapses stuck in childhood. PMID- 19812568 TI - Gene copy number: learning to count past two. PMID- 19812570 TI - Bleeding in the brain: Killer waves of depolarization in subarachnoid bleed. PMID- 19812571 TI - Bleeding in the brain: Amyloid-beta may keep clots away. PMID- 19812573 TI - Lasker Awards and papal portraiture: turning fields upside down. PMID- 19812574 TI - Nuclear reprogramming in eggs. PMID- 19812575 TI - Ekiden to iPS Cells. PMID- 19812576 TI - Perspectives on the development of imatinib and the future of cancer research. PMID- 19812577 TI - Attacking cancer at its foundation. PMID- 19812578 TI - Shifting paradigms: the seeds of oncogene addiction. PMID- 19812579 TI - What does the retrosplenial cortex do? AB - The past decade has seen a transformation in research on the retrosplenial cortex (RSC). This cortical area has emerged as a key member of a core network of brain regions that underpins a range of cognitive functions, including episodic memory, navigation, imagination and planning for the future. It is now also evident that the RSC is consistently compromised in the most common neurological disorders that impair memory. Here we review advances on multiple fronts, most notably in neuroanatomy, animal studies and neuroimaging, that have highlighted the importance of the RSC for cognition, and consider why specifying its precise functions remains problematic. PMID- 19812580 TI - Nocturnal penile erections: the diagnostic value of tumescence and rigidity activity units. AB - One of the most commonly used tests to distinguish psychogenic from organic erectile dysfunction (ED) is to monitor nocturnal penile tumescence using the RigiScan device and its new software called RigiScan Plus. To give a true estimate of the predictive ability of the new RigiScan software parameters, tumescence activity units (TAUs) and rigidity activity units (RAUs), we conducted this study on 639 RigiScan night records of 416 ED patients. For study purposes, these records were transferred to a personal computer and classified as normal and abnormal. We recorded the TAU and RAU provided by the RigiScan software for each event separately and also for the total night. We then estimated the diagnostic performance of these two parameters using cutoff values with highest accuracy plotted against the previously reported normal and abnormal curves. We then made four new calculations to improve the diagnostic accuracy of TAU and RAU for the total night. On estimating the highest diagnostic accuracy of RAU and TAU, it ranged from 67.8 to 73.7% for the single best event and from 68.4 to 74.2% for the total night. When using the newly computed units, the highest diagnostic accuracy did not exceed 75.9%. PMID- 19812582 TI - Repeated tapping of ventricular reservoir in preterm infants with post hemorrhagic ventricular dilatation does not increase the risk of reservoir infection. AB - OBJECTIVE: Controlled removal of cerebrospinal fluid (CSF) by serial tapping of ventricular reservoir, such as the McComb reservoir, is an effective way to decompress the ventricular system in preterm infants with rapidly progressing post-hemorrhagic ventricular dilatation (PHVD) while awaiting optimal conditions for permanent CSF drainage through a ventriculo-peritoneal shunt. However, the data regarding the risk of infection from repeated invasive tapping of a ventricular reservoir over a prolonged period are scarce. The aim of this study is to determine the incidence of ventricular reservoir infection from repeated tapping and to evaluate how often reservoir infection accompanies blood culture proven sepsis in preterm infants with PHVD. STUDY DESIGN: We reviewed the medical records of all infants with PHVD receiving serial reservoir tap at the University of Michigan from January 2000 through June 2007. Serial reservoir taps were carried out by the neonatology team using aseptic technique. Surveillance CSF cultures were sent on a weekly basis as per unit practice or when clinical sepsis was suspected. Reservoir-tapping-related infection was present if a reservoir became infected during serial tapping 7 days or more after surgical placement of the reservoir. RESULT: During the study period, ventricular reservoirs were placed in 35 infants for management of hydrocephalus. Six infants (17%) received ventricular reservoirs secondary to post-meningitic hydrocephalus or congenital brain malformations and were excluded. In the remaining 29 infants (birth weight: 1070+/-639 g (range 525 to 3204); gestational age: 26.9+/-4.1 weeks (range 23 to 40)) with PHVD, serial tapping of the ventricular reservoir was performed on 681 occasions (average number of taps per infant: 24 (range 2 to 82)). There were no cases of CSF culture-proven reservoir infection related to repeated taps. Thirteen of the 29 (45%) infants with PHVD developed blood culture-proven late onset sepsis with the following organisms: coagulase-negative Staphylococcus (9), Candida albicans (2), Escherichia coli (1), and Staphylococcus aureus (1), but all of the accompanying CSF cultures from the reservoir were negative. CONCLUSION: Ventricular reservoir infection from serial taps neither occurred in this consecutive series of preterm infants with PHVD, nor did ventricular reservoir infection accompany blood culture-proven sepsis. Concern of reservoir infection from repeated tapping should not be a limiting factor against placement of reservoirs. PMID- 19812581 TI - Inhaled nitric oxide in premature infants: effect on tracheal aspirate and plasma nitric oxide metabolites. AB - OBJECTIVE: Inhaled nitric oxide (iNO) is a potential new therapy for prevention of bronchopulmonary dysplasia and brain injury in premature infants. This study examined dose-related effects of iNO on NO metabolites as evidence of NO delivery. STUDY DESIGN: A subset of 102 premature infants in the NO CLD trial, receiving 24 days of iNO (20 p.p.m. decreasing to 2 p.p.m.) or placebo, were analyzed. Tracheal aspirate (TA) and plasma samples collected at enrollment and at intervals during study gas were analyzed for NO metabolites. RESULT: iNO treatment increased NO metabolites in TA at 20 and 10 p.p.m. (1.7- to 2.3-fold vs control) and in plasma at 20, 10, and 5 p.p.m. (1.6- to 2.3-fold). In post hoc analysis, treated infants with lower metabolite levels at entry had an improved clinical outcome. CONCLUSION: iNO causes dose-related increases in NO metabolites in the circulation as well as lung fluid, as evidenced by TA analysis, showing NO delivery to these compartments. PMID- 19812583 TI - Hospital neonatal services in the United States: variation in definitions, criteria, and regulatory status, 2008. AB - OBJECTIVE: The purpose of this study was to describe variation among states in designations of hospital neonatal services levels. STUDY DESIGN: We systematically searched all 50 states and District of Columbia governmental web sites and extracted definitions and levels terminology, functional and utilization criteria, regulatory compliance and funding measures, and citation of American Academy of Pediatrics (AAP) documents on levels of neonatal care. RESULT: Thirty-three states designate multiple graduated levels of neonatal services. Two to six levels were designated by numbers, titles, or both. Regulatory sources include hospital licensure, Certificate of Need or State Health Plan (CON/SHP), State Health Department, or an affiliated non-governmental entity (SHD/affiliate). Twenty-four states have a single source and nine have two or more. Functional criteria include population characteristics, respiratory care capabilities, and neonatal and cardiac surgery in 25 states. Utilization criteria include capacity, volume, occupancy, or case mix. Compliance mechanisms include license renewal, CON/SHP approval, and/or SHD/affiliate certification. Thirteen states link funding for the highest level of care through Medicaid, Maternal Child Health Title V funds or regional programs. AAP documents are cited or incorporated by reference in 22 states. CONCLUSION: All states regulate health care services and facilities. Definitions, criteria, compliance mechanisms, and regulatory source and status of neonatal levels of service vary widely. A consistent national approach would facilitate comparisons in neonatal outcomes and resource use and be informative to parents, providers, and policy makers. AAP documents could serve as a mechanism to foster such consistency. PMID- 19812584 TI - Pneumatoceles in preterm infants-incidence and outcome in the post-surfactant era. AB - OBJECTIVE: Pneumatoceles are gas-filled cysts within the lung parenchyma resulting mostly from ventilator-induced lung injury and air-leak in premature infants with respiratory distress syndrome. The use of surfactant in the treatment of respiratory distress syndrome has resulted in a decrease in the incidence of air-leak disease. Our aim was to study the incidence and clinical course of pneumatoceles in the surfactant era. STUDY DESIGN: A retrospective study of infants born at < or =30 weeks gestational age was admitted to the University of Connecticut Health Center NICU from 1998 to 2007. Pneumatoceles and other intrathoracic air-leaks were identified and comparisons were made with infants without these conditions. RESULT: Pneumatoceles were identified in 19 preterm infants, born at gestational age < or =30 weeks, needing positive pressure ventilation for respiratory distress syndrome between the years 1998 to 2007. Pneumatoceles appeared early (median, 7th day of life; range, 1st to 28th day of life) and usually resolved with decrease in mean airway pressure (median, 4 days; range, 3 to 125 days). The majority of pneumatoceles were located in the right parahilar region (18/19). Associated intrathoracic air-leaks were pulmonary interstitial emphysema (5/19), pneumothorax (10/19), and pneumomediastinum (1/19). None of the infants required any invasive procedures to alleviate the pneumatoceles. In infants who survived, most pneumatoceles resolved with a decrease in mean airway pressure or extubation (14/15). One infant had a persistent pneumatocele for 125 days without any cardiopulmonary compromise and five infants died as a result air-leaks along with other complications of prematurity. CONCLUSION: Pneumatoceles are a manifestation of intrathoracic air leaks of prematurity. They are markers for ventilator-induced lung injury and are associated with significant mortality similar to other intrathoracic air-leaks. However, conservative management with reduction in mean airway pressure is effective in the resolution of this condition and interventional decompression of the pneumatocele is generally not necessary. PMID- 19812585 TI - Failure of pediatric and neonatal trainees to meet Canadian Neonatal Resuscitation Program standards for neonatal intubation. AB - OBJECTIVE: Neonatal intubation skills are initially taught through the Neonatal Resuscitation Program (NRP) and thereafter complemented by further practical clinical training. The aim of this study is to compare the ability of NRP trained individuals to successfully complete a neonatal intubation. STUDY DESIGN: A prospective observational study was performed at an inborn high-risk level 3 perinatal center. Participants were postgraduate years 1 and 3 pediatric residents, neonatal-perinatal medicine subspecialty residents and fellows, and neonatal intensive care unit (NICU) respiratory therapists (RTs) with earlier NRP training. Intubations were scored on a checklist as well as a global assessment scale. Characteristics of the intubation attempt were recorded for each patient. RESULT: Fifty neonatal intubations were assessed, of which 73% of the attempts were deemed successful. A higher proportion of endotracheal tubes were successfully placed by RTs (100%, P<0.05), compared with both NICU fellows (69%) and pediatric residents (63%). The overall mean time for successful neonatal intubation was 51+/-28 s, which is greater than twice the time currently recommended by the NRP and American Heart Association guidelines. Attempts by pediatric residents and NICU fellows were longer (P<0.05, analysis of variance) and received lower global assessment scale (P<0.05, analysis of variance) and checklist (P<0.05, analysis of variance) scores, when compared with RTs. CONCLUSION: The success rate and overall quality of neonatal intubations performed by neonatal and pediatric trainees in Canada did not meet NRP standards; in particular, the time taken to intubate by pediatric residents and neonatal fellows is concerning. Re-evaluation of training methods and the volume of formalized exposure to neonatal intubation in Canadian residency programs are required. PMID- 19812586 TI - Prevalence of renal anomalies after urinary tract infections in hospitalized infants less than 2 months of age. AB - OBJECTIVE: Our aim was to determine the incidence of anatomical abnormalities after a urinary tract infection (UTI) in infants <2 months of age hospitalized in the neonatal intensive care unit (NICU). STUDY DESIGN: This was a retrospective, single-center cohort study of infants <2 months of age in the NICU with a UTI and documented renal imaging. RESULT: We identified 141 infants with UTIs. The mean gestational age and birth weight were 28 weeks and 1254 g, respectively. The most commonly identified pathogen was coagulase-negative Staphylococcus (28%, 44 of 156). A major abnormality was found on at least one imaging study for 4% (5 of 118) of infants. Major abnormalities were noted on 4% (5 of 114) of renal ultrasounds and 2% (2 of 82) of voiding cystourethrography examinations. CONCLUSION: Among infants in the NICU <2 months of age at the time of a UTI, the prevalence of major anatomical abnormalities is <5%. PMID- 19812587 TI - Pilot trial to compare tolerance of chlorhexidine gluconate to povidone-iodine antisepsis for central venous catheter placement in neonates. AB - OBJECTIVE: The purpose of this pilot trial was to determine whether rates of contact dermatitis following cutaneous antisepsis for central catheter placement were similar among neonates treated with chlorhexidine gluconate and povidone iodine. Chlorhexidine gluconate absorption was also evaluated. STUDY DESIGN: Infants weighing > or =1500 g and > or =7 days of age were randomized to a 10% povidone-iodine or 2% chlorhexidine gluconate site scrub before catheter placement. Primary outcomes evaluated included dermatitis, catheter colonization and chlorhexidine gluconate absorption. RESULT: A total of 48 neonates were enrolled. Colonization rates were similar among treatment groups (P<0.6). Dermatitis did not occur at chlorhexidine gluconate (central catheters, n=24; peripheral catheters, n=29) sites. Seven neonates had measurable chlorhexidine gluconate concentrations (range 13 to 100 ng ml(-1)) during catheterization. CONCLUSION: In this small trial chlorhexidine gluconate antisepsis was tolerated by study neonates. Chlorhexidine gluconate was cutaneously absorbed. Larger trials are needed to determine efficacy and tolerance of chlorhexidine gluconate in neonates. PMID- 19812588 TI - Pertussis antibodies in postpartum women and their newborns. AB - OBJECTIVE: To (1) determine the proportion of mothers and infants who had levels of IgG antibody to pertussis antigens predicted to be potentially protective at delivery; (2) evaluate the efficiency of maternal-infant antibody transport; (3) extrapolate infant antibody titers at 6 weeks; and (4) identify maternal factors associated with potentially protective infant antibodies. STUDY DESIGN: Sera from mother-infant pairs from February 2006 through to April 2007 were tested for antibody to pertussis antigens by standardized ELISA (enzyme-linked immunosorbent assay). Potentially protective antibody levels were defined as >5 ELISA units (EU) for pertussis toxin (PT), and >10 EU for fimbriae (FIM) and pertactin (PRN). Serological evidence of previous maternal infection was defined from antibody to four antigens by k-means cluster analysis. RESULT: In total, 21% (17/81) of mothers and 26% (21/81) of infants had potentially protective antibody levels at delivery. Mean infant-maternal antibody ratios for PT, FIM and PRN were 1.26, 1.36 and 1.31, respectively. At 6 weeks, 11% (9/81) of infants were predicted to have potentially protective antibody levels. Using cluster analysis, 9% (7/81) of mothers had evidence of previous pertussis infection. Infants born to these mothers were predicted to be more likely to have potentially protective antibodies at 6 weeks (43%) than those born to mothers without previous infection (8%) (P=0.03). CONCLUSION: Approximately 75% of infants were born with pertussis antibody levels lower than the modest levels associated with potential protection. Despite effective antibody transfer, nearly 90% of infants were predicted to have little antibody by 6 weeks. Maternal immunization before or during pregnancy might simulate previous pertussis infection and help protect infants through the first months of life. PMID- 19812589 TI - Impact of prematurity and co-morbidities on feeding milestones in neonates: a retrospective study. AB - OBJECTIVE: Feeding problems are an important area of neonatal morbidity that requires attention. We defined the feeding milestones related to transition to per oral feeding among premature infants based on gestational (GA) and postmenstrual ages (PMA), and elucidated the co-morbidity variables affecting with these skills. STUDY DESIGN: Feeding progress was tracked during the first hospitalization in a retrospective study involving 186 infants. We measured the age at acquisition of first feedings, maximum gavage feedings and maximum oral feedings. Resource usage measures included the total length of hospital stay (LOS), duration of gavage tube and duration of respiratory support. Effects of perinatal and co-morbidity factors on the acquisition of feeding milestones were evaluated. ANOVA, t-test, Wilcoxon rank sum test, chi(2) test, univariate and multivariate analysis, stepwise linear regression analysis and logistic regression analysis were applied as appropriate. Data were presented as mean+/ s.d., or as stated. P<0.05 was considered significant. RESULT: We stratified the data into three groups based on GA at birth: <28.0 weeks (group-1), 28.0 to 32.0 weeks (group-2) and 32.0 to 35.0 weeks (group-3). Compared with group-3, group-1 needed four-fold more ventilation and five-fold more continuous positive airway pressure (CPAP) duration (all P<0.001); whereas group-2 needed two-fold more CPAP duration. Age at first feed correlated with age at full gavage feedings and age at full oral feedings (r=0.53 and r=0.71, both P<0.0001). Age at full gavage feedings correlated with age at full oral feedings (r=0.81, P<0.0001). Univariate analysis was significant for GA age, hypotension, the effects of gastroesophageal reflux, and duration of ventilation and CPAP on the PMA at maximal oral feedings (all P<0.05); multivariate analysis for these variables was also significant (R (2)=0.58, P<0.0001). The success rate for oral feedings at discharge accelerated with GA maturation and caffeine use; on the other hand, the need for respiratory support and management of positive blood culture were associated with failure rates (P<0.05). CONCLUSION: Infants < 28 weeks GA have significant feeding delays with respect to initiation and progression to maximal gavage and oral feedings, as well as prolonged LOS. Infants >28 weeks GA attained successful feeding milestones by similar PMA. Specific aero-digestive co-morbidities significantly affected maximal oral feeding milestone. Delays in achieving maximum gavage and maximum oral feeding milestones suggest delays with the development of control and regulation of foregut motility. PMID- 19812590 TI - Adolescent mothers in the NICU: how much do they understand? AB - OBJECTIVE: We aimed to characterize adolescent parents' understanding of their infant's diagnosis, treatment and illness severity in the intensive care unit. STUDY DESIGN: Adolescent mothers were interviewed and neonatal medical records were reviewed. RESULT: Forty-two teens were interviewed. All had spoken with providers: 86% with nurses, 60% with physicians and 45% with both. Most teens could name their infant's diagnosis and treatment but often underestimated the illness severity. Teens reported reluctance to ask providers to clarify technical language. Those who said they spoke with a physician were less likely to understand their infant's illness severity than those who said they had not spoken with a physician (48 vs 82%). Parents' knowledge was better if physicians had documented explicit efforts to communicate with parents. CONCLUSION: Teens often underestimated the critical nature of their infant's illness. Future work should target adolescent willingness to ask questions and provider ability to accurately gauge parent knowledge. PMID- 19812591 TI - Neonatal outcomes associated with planned vaginal versus planned primary cesarean delivery. AB - OBJECTIVE: To determine whether planned route of delivery leads to differences in neonatal morbidity. STUDY DESIGN: Analysis was based on planned route of delivery, not actual route of delivery. A total of 4048 subjects were divided into two groups: planned vaginal delivery and planned cesarean delivery. Primary outcomes were neonatal intensive care unit (NICU) admission, respiratory morbidity and neurologic morbidity. RESULT: There were 3868 planned vaginal and 180 planned cesarean deliveries. Planned vaginal delivery had decreased NICU admission (P<0.0001), oxygen resuscitation (P=0.001) and jaundice (P<0.0001) but increased meconium passage (P<0.0001) and 1 min Apgar 43-fold coverage) and transcriptomes of an oestrogen-receptor-alpha positive metastatic lobular breast cancer at depth. We found 32 somatic non synonymous coding mutations present in the metastasis, and measured the frequency of these somatic mutations in DNA from the primary tumour of the same patient, which arose 9 years earlier. Five of the 32 mutations (in ABCB11, HAUS3, SLC24A4, SNX4 and PALB2) were prevalent in the DNA of the primary tumour removed at diagnosis 9 years earlier, six (in KIF1C, USP28, MYH8, MORC1, KIAA1468 and RNASEH2A) were present at lower frequencies (1-13%), 19 were not detected in the primary tumour, and two were undetermined. The combined analysis of genome and transcriptome data revealed two new RNA-editing events that recode the amino acid sequence of SRP9 and COG3. Taken together, our data show that single nucleotide mutational heterogeneity can be a property of low or intermediate grade primary breast cancers and that significant evolution can occur with disease progression. PMID- 19812675 TI - Structural insights into mechanisms of the small RNA methyltransferase HEN1. AB - RNA silencing is a conserved regulatory mechanism in fungi, plants and animals that regulates gene expression and defence against viruses and transgenes. Small silencing RNAs of approximately 20-30 nucleotides and their associated effector proteins, the Argonaute family proteins, are the central components in RNA silencing. A subset of small RNAs, such as microRNAs and small interfering RNAs (siRNAs) in plants, Piwi-interacting RNAs in animals and siRNAs in Drosophila, requires an additional crucial step for their maturation; that is, 2'-O methylation on the 3' terminal nucleotide. A conserved S-adenosyl-l-methionine dependent RNA methyltransferase, HUA ENHANCER 1 (HEN1), and its homologues are responsible for this specific modification. Here we report the 3.1 A crystal structure of full-length HEN1 from Arabidopsis in complex with a 22-nucleotide small RNA duplex and cofactor product S-adenosyl-l-homocysteine. Highly cooperative recognition of the small RNA substrate by multiple RNA binding domains and the methyltransferase domain in HEN1 measures the length of the RNA duplex and determines the substrate specificity. Metal ion coordination by both 2' and 3' hydroxyls on the 3'-terminal nucleotide and four invariant residues in the active site of the methyltransferase domain suggests a novel Mg(2+)-dependent 2'-O-methylation mechanism. PMID- 19812680 TI - The first transannular [4+3] cycloaddition reaction: synthesis of the ABCD ring structure of cortistatins. AB - A 14-membered macrocycle with an allene and a furan strategically located at across the ring from each other is synthesized using an allene ring closing metathesis reaction. Upon treatment of the macrocycle with a catalytic amount of Pd(OAc)(2) and other additives, the first transannular [4+3] cycloaddition occurred to yield 37% of a tetracyclic compound containing the ABC ring structure of the natural products cortistatins. PMID- 19812673 TI - A genome-wide linkage and association scan reveals novel loci for autism. AB - Although autism is a highly heritable neurodevelopmental disorder, attempts to identify specific susceptibility genes have thus far met with limited success. Genome-wide association studies using half a million or more markers, particularly those with very large sample sizes achieved through meta-analysis, have shown great success in mapping genes for other complex genetic traits. Consequently, we initiated a linkage and association mapping study using half a million genome-wide single nucleotide polymorphisms (SNPs) in a common set of 1,031 multiplex autism families (1,553 affected offspring). We identified regions of suggestive and significant linkage on chromosomes 6q27 and 20p13, respectively. Initial analysis did not yield genome-wide significant associations; however, genotyping of top hits in additional families revealed an SNP on chromosome 5p15 (between SEMA5A and TAS2R1) that was significantly associated with autism (P = 2 x 10(-7)). We also demonstrated that expression of SEMA5A is reduced in brains from autistic patients, further implicating SEMA5A as an autism susceptibility gene. The linkage regions reported here provide targets for rare variation screening whereas the discovery of a single novel association demonstrates the action of common variants. PMID- 19812681 TI - The catalytic asymmetric addition of alkyl- and aryl-zinc reagents to an isoxazole aldehyde. AB - Nucleophilic addition of alkyl- and aryl zinc reagents to a C(4) functionalized isoxazolyl aldehyde proceeded effectively with high enantioselectivity (85-94% e.e.). The amino alcohol catalyst (S)-2-piperidinyl-1,1,2-triphenyl ethanol (10) afforded the (R)-product 2b, as established by x-ray crystallography. PMID- 19812682 TI - Multiple chirality transfers in the enantioselective synthesis of 11-O debenzoyltashironin. Chiroptical analysis of the key cascade. AB - The mechanism of the cascade oxidative dearomatization-transannular Diels Alder was investigated in the context of an asymmetric route to (-)-11-O debenzoyltashironin. Although the oxidative dearomatization provides two acetal intermediates, the transannular Diels-Alder proceeds spontaneously from only one of the acetal isomers. Access to enantioenriched tetracyclic adduct was gained through the use of optically active allene. PMID- 19812683 TI - Exteriorization of buried port to salvage infected tissue expander. AB - OBJECTIVE: Since removal of an infected tissue expander is very disappointing to both the surgeon and the patient, every effort is directed toward its salvage. This study evaluates a new method to salvage infected tissue expanders. METHOD: Of 66 tissue expanders applied at different sites in the body, 12 developed infection. Salvage was carried out by exteriorizing the buried port, followed by irrigation through the pocket of the tube that connects the port to the expander. RESULT: Salvage was successful in 9 of the infected tissue expanders and failed in 3 cases. The ports were not dependent in these 3 cases. CONCLUSION: Exteriorization of dependent ports allows adequate drainage, good access for irrigation, and completion of expansion. PMID- 19812684 TI - Chronic obstructive pulmonary disease and altered risk of lung cancer in a population-based case-control study. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) has been consistently associated with increased risk of lung cancer. However, previous studies have had limited ability to determine whether the association is due to smoking. METHODOLOGY/PRINCIPAL FINDINGS: The Environment And Genetics in Lung cancer Etiology (EAGLE) population-based case-control study recruited 2100 cases and 2120 controls, of whom 1934 cases and 2108 controls reported about diagnosis of chronic bronchitis, emphysema, COPD (chronic bronchitis and/or emphysema), or asthma more than 1 year before enrollment. We estimated odds ratios (OR) and 95% confidence intervals (CI) using logistic regression. After adjustment for smoking, other previous lung diseases, and study design variables, lung cancer risk was elevated among individuals with a history of chronic bronchitis (OR = 2.0, 95% CI = 1.5-2.5), emphysema (OR = 1.9, 95% CI = 1.4-2.8), or COPD (OR = 2.5, 95% CI = 2.0-3.1). Among current smokers, association between chronic bronchitis and lung cancer was strongest among lighter smokers. Asthma was associated with a decreased risk of lung cancer in males (OR = 0.48, 95% CI = 0.30-0.78). CONCLUSIONS/SIGNIFICANCE: These results suggest that the associations of personal history of chronic bronchitis, emphysema, and COPD with increased risk of lung cancer are not entirely due to smoking. Inflammatory processes may both contribute to COPD and be important for lung carcinogenesis. PMID- 19812685 TI - Heritability of antibody isotype and subclass responses to Plasmodium falciparum antigens. AB - BACKGROUND: It is important to understand the extent to which genetic factors regulate acquired immunity to common infections. A classical twin study design is useful to estimate the heritable component of variation in measurable immune parameters. METHODOLOGY/PRINCIPAL FINDINGS: This study assessed the relative heritability of different plasma antibody isotypes and subclasses (IgG1, IgG2, IgG3, IgG4, IgM, IgA and IgE) naturally acquired to P. falciparum blood stage antigens AMA1, MSP1-19, MSP2 (two allelic types) and MSP3 (two allelic types). Separate analyses were performed on plasma from 213 pairs of Gambian adult twins, 199 child twin pairs sampled in a dry season when there was little malaria transmission, and another set of 107 child twin pairs sampled at the end of the annual wet season when malaria was common. There were significantly positive heritability (h(2)) estimates for 48% (20/42) of the specific antibody assays (for the seven isotypes and subclasses to the six antigens tested) among the adults, 48% (20/42) among the children in the dry season and 31% (13/42) among the children in the wet season. In children, there were significant heritability estimates for IgG4 reactivity against each of the antigens, and this subclass had higher heritability than the other subclasses and isotypes. In adults, 75% (15/20) of the significantly heritable antigen-specific isotype responses were attributable to non-HLA class II genetic variation, whereas none showed a significant HLA contribution. SIGNIFICANCE: Genome-wide approaches are now warranted to map the major genetic determinants of variable antibody isotype and subclass responses to malaria, alongside evaluation of their impact on infection and disease. Although plasma levels of IgG4 to malaria antigens are generally low, the exceptionally high heritability of levels of this subclass in children deserves particular investigation. PMID- 19812686 TI - Tumor-shed PGE(2) impairs IL2Rgammac-signaling to inhibit CD4 T cell survival: regulation by theaflavins. AB - BACKGROUND: Many tumors are associated with decreased cellular immunity and elevated levels of prostaglandin E2 (PGE2), a known inhibitor of CD4+ T cell activation and inducer of type-2 cytokine bias. However, the role of this immunomodulator in the survival of T helper cells remained unclear. Since CD4+ T cells play critical roles in cell-mediated immunity, detail knowledge of the effect tumor-derived PGE2 might have on CD4+ T cell survival and the underlying mechanism may, therefore, help to overcome the overall immune deviation in cancer. METHODOLOGY/PRINCIPAL FINDINGS: By culturing purified human peripheral CD4+ T cells or Jurkat cells with spent media of theaflavin- or celecoxib-pre treated MCF-7 cells, we show that tumor-shed PGE2 severely impairs interleukin 2 receptor gammac (IL2Rgammac)-mediated survival signaling in CD4+ T cells. Indeed, tumor-shed PGE2 down-regulates IL2Rgammac expression, reduces phosphorylation as well as activation of Janus kinase 3 (Jak-3)/signal transducer and activator of transcription 5 (Stat-5) and decreases Bcl-2/Bax ratio thereby leading to activation of intrinsic apoptotic pathway. Constitutively active Stat-5A (Stat 5A1 6) over-expression efficiently elevates Bcl-2 levels in CD4+ T cells and protects them from tumor-induced death while dominant-negative Stat-5A over expression fails to do so, indicating the importance of Stat-5A-signaling in CD4+ T cell survival. Further support towards the involvement of PGE2 comes from the results that (a) purified synthetic PGE2 induces CD4+ T cell apoptosis, and (b) when knocked out by small interfering RNA, cyclooxygenase-2 (Cox-2)-defective tumor cells fail to initiate death. Interestingly, the entire phenomena could be reverted back by theaflavins that restore cytokine-dependent IL2Rgammac/Jak 3/Stat-5A signaling in CD4+ T cells thereby protecting them from tumor-shed PGE2 induced apoptosis. CONCLUSIONS/SIGNIFICANCE: These data strongly suggest that tumor-shed PGE2 is an important factor leading to CD4+ T cell apoptosis during cancer and raise the possibility that theaflavins may have the potential as an effective immunorestorer in cancer-bearer. PMID- 19812687 TI - The amidase domain of lipoamidase specifically inactivates lipoylated proteins in vivo. AB - BACKGROUND: In the 1950s, Reed and coworkers discovered an enzyme activity in Streptococcus faecalis (Enterococcus faecalis) extracts that inactivated the Escherichia. coli and E. faecalis pyruvate dehydrogenase complexes through cleavage of the lipoamide bond. The enzyme that caused this lipoamidase activity remained unidentified until Jiang and Cronan discovered the gene encoding lipoamidase (Lpa) through the screening of an expression library. Subsequent cloning and characterization of the recombinant enzyme revealed that lipoamidase is an 80 kDa protein composed of an amidase domain containing a classic Ser-Ser Lys catalytic triad and a carboxy-terminal domain of unknown function. Here, we show that the amidase domain can be used as an in vivo probe which specifically inactivates lipoylated enzymes. METHODOLOGY/PRINCIPAL FINDINGS: We evaluated whether Lpa could function as an inducible probe of alpha-ketoacid dehydrogenase inactivation using E. coli as a model system. Lpa expression resulted in cleavage of lipoic acid from the three lipoylated proteins expressed in E. coli, but did not result in cleavage of biotin from the sole biotinylated protein, the biotin carboxyl carrier protein. When expressed in lipoylation deficient E. coli, Lpa is not toxic, indicating that Lpa does not interfere with any other critical metabolic pathways. When truncated to the amidase domain, Lpa retained lipoamidase activity without acquiring biotinidase activity, indicating that the carboxy-terminal domain is not essential for substrate recognition or function. Substitution of any of the three catalytic triad amino acids with alanine produced inactive Lpa proteins. CONCLUSIONS/SIGNIFICANCE: The enzyme lipoamidase is active against a broad range of lipoylated proteins in vivo, but does not affect the growth of lipoylation deficient E. coli. Lpa can be truncated to 60% of its original size with only a partial loss of activity, resulting in a smaller probe that can be used to study the effects of alpha-ketoacid dehydrogenase inactivation in vivo. PMID- 19812688 TI - The contribution of TRPM8 and TRPA1 channels to cold allodynia and neuropathic pain. AB - Cold allodynia is a common feature of neuropathic pain however the underlying mechanisms of this enhanced sensitivity to cold are not known. Recently the transient receptor potential (TRP) channels TRPM8 and TRPA1 have been identified and proposed to be molecular sensors for cold. Here we have investigated the expression of TRPM8 and TRPA1 mRNA in the dorsal root ganglia (DRG) and examined the cold sensitivity of peripheral sensory neurons in the chronic construction injury (CCI) model of neuropathic pain in mice.In behavioral experiments, chronic constriction injury (CCI) of the sciatic nerve induced a hypersensitivity to both cold and the TRPM8 agonist menthol that developed 2 days post injury and remained stable for at least 2 weeks. Using quantitative RT-PCR and in situ hybridization we examined the expression of TRPM8 and TRPA1 in DRG. Both channels displayed significantly reduced expression levels after injury with no change in their distribution pattern in identified neuronal subpopulations. Furthermore, in calcium imaging experiments, we detected no alterations in the number of cold or menthol responsive neurons in the DRG, or in the functional properties of cold transduction following injury. Intriguingly however, responses to the TRPA1 agonist mustard oil were strongly reduced.Our results indicate that injured sensory neurons do not develop abnormal cold sensitivity after chronic constriction injury and that alterations in the expression of TRPM8 and TRPA1 are unlikely to contribute directly to the pathogenesis of cold allodynia in this neuropathic pain model. PMID- 19812689 TI - Candidate vaccine sequences to represent intra- and inter-clade HIV-1 variation. AB - A likely key factor in the failure of a HIV-1 vaccine based on cytotoxic T lymphocytes (CTL) is the natural immunodominance of epitopes that fall in variable regions of the proteome, which both increases the chance of epitope sequence mismatch with the incoming challenge strain and replicates the pathogenesis of early CTL failure due to epitope escape mutation during natural infection. To identify potential vaccine sequences to focus the CTL response on highly conserved epitopes, the whole proteomes of HIV-1 clades A1, B, C, and D were assessed for Shannon entropy at each amino acid position. Highly conserved regions in Gag (cGag-1, Gag 148-214, and cGag-2, Gag 253-331), Env (cEnv, Env 521 606), and Nef (cNef, Nef 106-148) were identified across clades. Inter- and intra clade variability of amino acids within the regions tended to overlap, suggesting that polyvalent representation of consensus sequences for the four clades would allow broad HIV-1 strain representation. These four conserved regions were rich in both known and predicted CTL epitopes presented by a breadth of HLA types, and screening of 54 persons with chronic HIV-1 infection revealed that these regions are commonly immunogenic in the context of natural infection. These data suggest that vaccine delivery of a 16-valent mixture of these regions could focus the CTL response against conserved epitopes that are broadly representative of circulating HIV-1 strains. PMID- 19812690 TI - Successful small intestine colonization of adult mice by Vibrio cholerae requires ketamine anesthesia and accessory toxins. AB - Vibrio cholerae colonizes the small intestine of adult C57BL/6 mice. In this study, the physical and genetic parameters that facilitate this colonization were investigated. Successful colonization was found to depend upon anesthesia with ketamine-xylazine and neutralization of stomach acid with sodium bicarbonate, but not streptomycin treatment. A variety of common mouse strains were colonized by O1, O139, and non-O1/non-O139 strains. All combinations of mutants in the genes for hemolysin, the multifunctional, autoprocessing RTX toxin (MARTX), and hemagglutinin/protease were assessed, and it was found that hemolysin and MARTX are each sufficient for colonization after a low dose infection. Overall, this study suggests that, after intragastric inoculation, V. cholerae encounters barriers to infection including an acidic environment and an immediate immune response that is circumvented by sodium bicarbonate and the anti-inflammatory effects of ketamine-xylazine. After initial adherence in the small intestine, the bacteria are subjected to additional clearance mechanisms that are evaded by the independent toxic action of hemolysin or MARTX. Once colonization is established, it is suggested that, in humans, these now persisting bacteria initiate synthesis of the major virulence factors to cause cholera disease. This adult mouse model of intestinal V. cholerae infection, now well-characterized and fully optimized, should serve as a valuable tool for studies of pathogenesis and testing vaccine efficacy. PMID- 19812691 TI - Thinking beyond low-density lipoprotein cholesterol: strategies to further reduce cardiovascular risk. AB - Several large statin trials and meta-analyses have demonstrated a reduction in low-density lipoprotein cholesterol (LDL-C) and cardiovascular morbidity and mortality. Some trials have also highlighted the significance of residual cardiovascular risk after treatment of LDL-C to target levels. This reflects the complex nature of residual cardiovascular risk. This residual risk is partially due to low HDL-C and high triglycerides (TG) despite achievement of LDL goals with statin therapy. The NCEP ATP III guidelines reported that low HDL-C is a significant and an independent risk factor for coronary heart disease (CHD) and is inversely related to CHD. Epidemiologic studies have also shown a similar inverse relationship of HDL-C with CHD. High-density lipoprotein cholesterol (HDL C) may directly participate in the anti-atherogenic process by promoting efflux of cholesterol of the foam cells of atherogenic lesions. Many studies have demonstrated multiple anti-atherogenic actions of HDL-C and its role in promoting efflux of cholesterol from the foam cells. The residual risk by increased TG with or without low HDL-C can be assessed by calculating non-HDL-C and a reduction in TG results in decreased CHD. PMID- 19812692 TI - Omega-3 polyunsaturated fatty acids: a necessity for a comprehensive secondary prevention strategy. AB - Long-chain omega-3 polyunsaturated fatty acid (PUFA) supplementation has been used for the secondary prevention of fatal and nonfatal myocardial infarction (MI). However, the benefit of this therapy is frequently confused with other established treatments in the therapeutic strategy among such patients. We review the data on omega-3 PUFA use in secondary care and consider indications for its use which include post-MI and raised triglycerides. We suggest that the available evidence supports the use of omega-3 supplementation as part of the comprehensive secondary care package for post-MI patients. PMID- 19812693 TI - Comparison between automated and manual measurements of carotid intima-media thickness in clinical practice. AB - BACKGROUND AND AIM: The measurement of carotid intima-media thickness (cIMT) has been used as a marker of arterial wall disease. Manual measurements have been performed in most epidemiological studies, but, due to the introduction of new technologies, automated software has been increasingly used. This study aimed to compare manual versus automated cIMT measurements in common carotid (CC), bifurcation (BIF), and internal carotid (IC). METHODS: Automated and manual cIMT measurements were performed online in 43 middle-aged females. Carotid segment measurements were compared by Bland-Altman plot and the variation and repeatability coefficients between observers were also determined for comparison. RESULTS: The average timespan for manual measurements (57.30 s) were significantly higher than for automated measurements (2.52 s). There were no systematic errors between methods in any carotid segments. The variation coefficient was 5.54% to 6.34% for CC and BIF, 9.76% for IC, and absolute differences were 85% below 0.1 mm and 70% below 0.05 mm. Interobserver agreement showed no systematic error. The variation and the repeatability coefficients were better for the automated than manual measures. CONCLUSION: Although both methods are reliable for cIMT measurements, the automated technique allows faster evaluation with lesser variability for all carotid segments currently used in atherosclerosis research. PMID- 19812694 TI - Integrating the genetic and physical maps of Arabidopsis thaliana: identification of mapped alleles of cloned essential (EMB) genes. AB - The classical genetic map of Arabidopsis includes more than 130 genes with an embryo-defective (emb) mutant phenotype. Many of these essential genes remain to be cloned. Hundreds of additional EMB genes have been cloned and catalogued (www.seedgenes.org) but not mapped. To facilitate EMB gene identification and assess the current level of saturation, we updated the classical map, compared the physical and genetic locations of mapped loci, and performed allelism tests between mapped (but not cloned) and cloned (but not mapped) emb mutants with similar chromosome locations. Two hundred pairwise combinations of genes located on chromosomes 1 and 5 were tested and more than 1100 total crosses were screened. Sixteen of 51 mapped emb mutants examined were found to be disrupted in a known EMB gene. Alleles of a wide range of published EMB genes (YDA, GLA1, TIL1, AtASP38, AtDEK1, EMB506, DG1, OEP80) were discovered. Two EMS mutants isolated 30 years ago, T-DNA mutants with complex insertion sites, and a mutant with an atypical, embryo-specific phenotype were resolved. The frequency of allelism encountered was consistent with past estimates of 500 to 1000 EMB loci. New EMB genes identified among mapped T-DNA insertion mutants included CHC1, which is required for chromatin remodeling, and SHS1/AtBT1, which encodes a plastidial nucleotide transporter similar to the maize Brittle1 protein required for normal endosperm development. Two classical genetic markers (PY, ALB1) were identified based on similar map locations of known genes required for thiamine (THIC) and chlorophyll (PDE166) biosynthesis. The alignment of genetic and physical maps presented here should facilitate the continued analysis of essential genes in Arabidopsis and further characterization of a broad spectrum of mutant phenotypes in a model plant. PMID- 19812695 TI - Nanoparticle-delivered multimeric soluble CD40L DNA combined with Toll-Like Receptor agonists as a treatment for melanoma. AB - Stimulation of CD40 or Toll-Like Receptors (TLR) has potential for tumor immunotherapy. Combinations of CD40 and TLR stimulation can be synergistic, resulting in even stronger dendritic cell (DC) and CD8+ T cell responses. To evaluate such combinations, established B16F10 melanoma tumors were injected every other day X 5 with plasmid DNA encoding a multimeric, soluble form of CD40L (pSP-D-CD40L) either alone or combined with an agonist for TLR1/2 (Pam(3)CSK(4) ), TLR2/6 (FSL-1 and MALP2), TLR3 (polyinosinic-polycytidylic acid, poly(I:C)), TLR4 ( monophosphoryl lipid A, MPL), TLR7 (imiquimod), or TLR9 (Class B CpG phosphorothioate oligodeoxynucleotide, CpG). When used by itself, pSP-D-CD40L slowed tumor growth and prolonged survival, but did not lead to cure. Of the TLR agonists, CpG and poly(I:C) also slowed tumor growth, and the combination of these two TLR agonists was more effective than either agent alone. The triple combination of intratumoral pSP-D-CD40L + CpG + poly(I:C) markedly slowed tumor growth and prolonged survival. This treatment was associated with a reduction in intratumoral CD11c+ dendritic cells and an influx of CD8+ T cells. Since intratumoral injection of plasmid DNA does not lead to efficient transgene expression, pSP-D-CD40L was also tested with cationic polymers that form DNA containing nanoparticles which lead to enhanced intratumoral gene expression. Intratumoral injections of pSP-D-CD40L-containing nanoparticles formed from polyethylenimine (PEI) or C32 (a novel biodegradable poly(B-amino esters) polymer) in combination with CpG + poly(I:C) had dramatic antitumor effects and frequently cured mice of B16F10 tumors. These data confirm and extend previous reports that CD40 and TLR agonists are synergistic and demonstrate that this combination of immunostimulants can significantly suppress tumor growth in mice. In addition, the enhanced effectiveness of nanoparticle formulations of DNA encoding immunostimulatory molecules such as multimeric, soluble CD40L supports the further study of this technology for tumor immunotherapy. PMID- 19812697 TI - Sour ageusia in two individuals implicates ion channels of the ASIC and PKD families in human sour taste perception at the anterior tongue. AB - BACKGROUND: The perception of sour taste in humans is incompletely understood at the receptor cell level. We report here on two patients with an acquired sour ageusia. Each patient was unresponsive to sour stimuli, but both showed normal responses to bitter, sweet, and salty stimuli. METHODS AND FINDINGS: Lingual fungiform papillae, containing taste cells, were obtained by biopsy from the two patients, and from three sour-normal individuals, and analyzed by RT-PCR. The following transcripts were undetectable in the patients, even after 50 cycles of amplification, but readily detectable in the sour-normal subjects: acid sensing ion channels (ASICs) 1a, 1beta, 2a, 2b, and 3; and polycystic kidney disease (PKD) channels PKD1L3 and PKD2L1. Patients and sour-normals expressed the taste related phospholipase C-beta2, the delta-subunit of epithelial sodium channel (ENaC) and the bitter receptor T2R14, as well as beta-actin. Genomic analysis of one patient, using buccal tissue, did not show absence of the genes for ASIC1a and PKD2L1. Immunohistochemistry of fungiform papillae from sour-normal subjects revealed labeling of taste bud cells by antibodies to ASICs 1a and 1beta, PKD2L1, phospholipase C-beta2, and delta-ENaC. An antibody to PKD1L3 labeled tissue outside taste bud cells. CONCLUSIONS: These data suggest a role for ASICs and PKDs in human sour perception. This is the first report of sour ageusia in humans, and the very existence of such individuals ("natural knockouts") suggests a cell lineage for sour that is independent of the other taste modalities. PMID- 19812696 TI - Cancer genomics identifies regulatory gene networks associated with the transition from dysplasia to advanced lung adenocarcinomas induced by c-Raf-1. AB - BACKGROUND: Lung cancer is a leading cause of cancer morbidity. To improve an understanding of molecular causes of disease a transgenic mouse model was investigated where targeted expression of the serine threonine kinase c-Raf to respiratory epithelium induced initially dysplasia and subsequently adenocarcinomas. This enables dissection of genetic events associated with precancerous and cancerous lesions. METHODOLOGY/PRINCIPAL FINDINGS: By laser microdissection cancer cell populations were harvested and subjected to whole genome expression analyses. Overall 473 and 541 genes were significantly regulated, when cancer versus transgenic and non-transgenic cells were compared, giving rise to three distinct and one common regulatory gene network. At advanced stages of tumor growth predominately repression of gene expression was observed, but genes previously shown to be up-regulated in dysplasia were also up-regulated in solid tumors. Regulation of developmental programs as well as epithelial mesenchymal and mesenchymal endothelial transition was a hall mark of adenocarcinomas. Additionally, genes coding for cell adhesion, i.e. the integrins and the tight and gap junction proteins were repressed, whereas ligands for receptor tyrosine kinase such as epi- and amphiregulin were up-regulated. Notably, Vegfr- 2 and its ligand Vegfd, as well as Notch and Wnt signalling cascades were regulated as were glycosylases that influence cellular recognition. Other regulated signalling molecules included guanine exchange factors that play a role in an activation of the MAP kinases while several tumor suppressors i.e. Mcc, Hey1, Fat3, Armcx1 and Reck were significantly repressed. Finally, probable molecular switches forcing dysplastic cells into malignantly transformed cells could be identified. CONCLUSIONS/SIGNIFICANCE: This study provides insight into molecular pertubations allowing dysplasia to progress further to adenocarcinoma induced by exaggerted c-Raf kinase activity. PMID- 19812698 TI - Population structure of humpback whales from their breeding grounds in the South Atlantic and Indian Oceans. AB - Although humpback whales are among the best-studied of the large whales, population boundaries in the Southern Hemisphere (SH) have remained largely untested. We assess population structure of SH humpback whales using 1,527 samples collected from whales at fourteen sampling sites within the Southwestern and Southeastern Atlantic, the Southwestern Indian Ocean, and Northern Indian Ocean (Breeding Stocks A, B, C and X, respectively). Evaluation of mtDNA population structure and migration rates was carried out under different statistical frameworks. Using all genetic evidence, the results suggest significant degrees of population structure between all ocean basins, with the Southwestern and Northern Indian Ocean most differentiated from each other. Effective migration rates were highest between the Southeastern Atlantic and the Southwestern Indian Ocean, followed by rates within the Southeastern Atlantic, and the lowest between the Southwestern and Northern Indian Ocean. At finer scales, very low gene flow was detected between the two neighbouring sub-regions in the Southeastern Atlantic, compared to high gene flow for whales within the Southwestern Indian Ocean. Our genetic results support the current management designations proposed by the International Whaling Commission of Breeding Stocks A, B, C, and X as four strongly structured populations. The population structure patterns found in this study are likely to have been influenced by a combination of long-term maternally directed fidelity of migratory destinations, along with other ecological and oceanographic features in the region. PMID- 19812699 TI - A man-made ATP-binding protein evolved independent of nature causes abnormal growth in bacterial cells. AB - Recent advances in de novo protein evolution have made it possible to create synthetic proteins from unbiased libraries that fold into stable tertiary structures with predefined functions. However, it is not known whether such proteins will be functional when expressed inside living cells or how a host organism would respond to an encounter with a non-biological protein. Here, we examine the physiology and morphology of Escherichia coli cells engineered to express a synthetic ATP-binding protein evolved entirely from non-biological origins. We show that this man-made protein disrupts the normal energetic balance of the cell by altering the levels of intracellular ATP. This disruption cascades into a series of events that ultimately limit reproductive competency by inhibiting cell division. We now describe a detailed investigation into the synthetic biology of this man-made protein in a living bacterial organism, and the effect that this protein has on normal cell physiology. PMID- 19812700 TI - Crosstalks between myo-inositol metabolism, programmed cell death and basal immunity in Arabidopsis. AB - BACKGROUND: Although it is a crucial cellular process required for both normal development and to face stress conditions, the control of programmed cell death in plants is not fully understood. We previously reported the isolation of ATXR5 and ATXR6, two PCNA-binding proteins that could be involved in the regulation of cell cycle or cell death. A yeast two-hybrid screen using ATXR5 as bait captured AtIPS1, an enzyme which catalyses the committed step of myo-inositol (MI) biosynthesis. atips1 mutants form spontaneous lesions on leaves, raising the possibility that MI metabolism may play a role in the control of PCD in plants. In this work, we have characterised atips1 mutants to gain insight regarding the role of MI in PCD regulation. METHODOLOGY/PRINCIPAL FINDINGS: - lesion formation in atips1 mutants depends of light intensity, is due to PCD as evidenced by TUNEL labelling of nuclei, and is regulated by phytohormones such as salicylic acid - MI and galactinol are the only metabolites whose accumulation is significantly reduced in the mutant, and supplementation of the mutant with these compounds is sufficient to prevent PCD - the transcriptome profile of the mutant is extremely similar to that of lesion mimic mutants such as cpr5, or wild-type plants infected with pathogens. CONCLUSION/SIGNIFICANCE: Taken together, our results provide strong evidence for the role of MI or MI derivatives in the regulation of PCD. Interestingly, there are three isoforms of IPS in Arabidopsis, but AtIPS1 is the only one harbouring a nuclear localisation sequence, suggesting that nuclear pools of MI may play a specific role in PCD regulation and opening new research prospects regarding the role of MI in the prevention of tumorigenesis. Nevertheless, the significance of the interaction between AtIPS1 and ATXR5 remains to be established. PMID- 19812701 TI - Adult reversal of cognitive phenotypes in neurodevelopmental disorders. AB - Recent findings in mice suggest that it is possible to reverse certain neurodevelopmental disorders in adults. Changes in development, previously thought to be irreparable in adults, were believed to underlie the neurological and psychiatric phenotypes of a range of common mental health problems with a clear developmental component. As a consequence, most researchers have focused their efforts on understanding the molecular and cellular processes that alter development with the hope that early intervention could prevent the emergent pathology. Unexpectedly, several different animal model studies published recently, including animal models of autism, suggest that it may be possible to reverse neurodevelopmental disorders in adults: Addressing the underlying molecular and cellular deficits in adults could in several cases dramatically improve the neurocognitive phenotypes in these animal models. The findings reviewed here provide hope to millions of individuals afflicted with a wide range of neurodevelopmental disorders, including autism, since they suggest that it may be possible to treat or even cure them in adults. PMID- 19812702 TI - Abnormal 18F-FDG Uptake Detected with Positron Emission Tomography in a Patient with Breast Cancer: A Case of Sarcoidosis and Review of the Literature. AB - 18F-FDG PET is a useful and sensitive imaging method for a variety of malignancies, however, the specificity is low in active infections and inflammatory diseases. We describe a female patient with stage IIIA breast cancer in first complete remission with combination chemotherapy who developed nodular formations in the lung and axilla 12 years later. Imaging studies as well as FDG PET showed nodular lesions and increased metabolic activity which was interpreted as the progression of the primary disease. She was first given combination chemotherapy and hormonal therapy but was proven thereafter to have sarcoidosis by pathologic examination and was successfully treated with corticosteroid treatment. PMID- 19812703 TI - Structural and ultrastructural analysis of cerebral cortex, cerebellum, and hypothalamus from diabetic rats. AB - Autonomic and peripheral neuropathies are well-described complications in diabetes. Diabetes mellitus is also associated to central nervous system damage. This little-known complication is characterized by impairment of brain functions and electrophysiological changes associated with neurochemical and structural abnormalities. The purpose of this study was to investigate brain structural and ultrastructural changes in rats with streptozotocin-induced diabetes. Cerebral cortex, hypothalamus, and cerebellum were obtained from controls and 8 weeks diabetic rats. Light and electron microscope studies showed degenerative changes of neurons and glia, perivascular and mitochondrial swelling, disarrangement of myelin sheath, increased area of myelinated axons, presynaptic vesicle dispersion in swollen axonal boutoms, fragmentation of neurofilaments, and oligodendrocyte abnormalities. In addition, depressive mood was observed in diabetic animals. The brain morphological alterations observed in diabetic animals could be related to brain pathologic process leading to abnormal function, cellular death, and depressive behavioral. PMID- 19812704 TI - Image restoration using functional and anatomical information fusion with application to SPECT-MRI images. AB - Image restoration is usually viewed as an ill-posed problem in image processing, since there is no unique solution associated with it. The quality of restored image closely depends on the constraints imposed of the characteristics of the solution. In this paper, we propose an original extension of the NAS-RIF restoration technique by using information fusion as prior information with application in SPECT medical imaging. That extension allows the restoration process to be constrained by efficiently incorporating, within the NAS-RIF method, a regularization term which stabilizes the inverse solution. Our restoration method is constrained by anatomical information extracted from a high resolution anatomical procedure such as magnetic resonance imaging (MRI). This structural anatomy-based regularization term uses the result of an unsupervised Markovian segmentation obtained after a preliminary registration step between the MRI and SPECT data volumes from each patient. This method was successfully tested on 30 pairs of brain MRI and SPECT acquisitions from different subjects and on Hoffman and Jaszczak SPECT phantoms. The experiments demonstrated that the method performs better, in terms of signal-to-noise ratio, than a classical supervised restoration approach using a Metz filter. PMID- 19812705 TI - The impact of combination antiretroviral therapy and its interruption on anxiety, stress, depression and quality of life in Thai patients. AB - OBJECTIVE: Investigation on anxiety, stress, depression, and quality of life (QoL) within STACCATO, a randomised trial of two treatment strategies: CD4 guided scheduled treatment interruption (STI) compared to continuous treatment (CT). PARTICIPANTS: Thai patients with HIV-infection enrolled in the STACCATO trial. METHODS: Anxiety, depression assessed by the questionnaires Hospital Anxiety and Depression Scale (HADS) and DASS, stress assessed by the Depression Anxiety Stress Scale (DASS), and QoL evaluated by the HIV Medical Outcome Study (MOS-HIV) questionnaires. Answers to questionnaires were evaluated at 4 time-points: baseline, 24 weeks, 48 weeks and at the end of STACCATO. RESULTS: A total of 251 patients answered the HADS/DASS and 241 answered the MOS-HIV of the 379 Thai patients enrolled into STACCATO (66.2 and 63.6% respectively). At baseline 16.3% and 7.2% of patients reported anxiety and depression using HADS scale. Using the DASS scale, 35.1% reported mild to moderate and 9.6% reported severe anxiety; 8.8% reported mild to moderate and 2.0% reported severe depression; 42.6% reported mild to moderate and 4.8% reported severe stress. We showed a significant improvement of the MHS across time (p=0.001), but no difference between arms (p=0.17). The summarized physical health status score (PHS) did not change during the trial (p=0.15) nor between arm (p=0.45). There was no change of MHS or PHS in the STI arm, taking into account the number of STI cycle (p=0.30 and 0.57) but MHS significant increased across time-points (p=0.007). CONCLUSION: Antiretroviral therapy improved mental health and QOL, irrespective of the treatment strategy. PMID- 19812706 TI - Deterministic chaos and fractal complexity in the dynamics of cardiovascular behavior: perspectives on a new frontier. AB - Physiological systems such as the cardiovascular system are capable of five kinds of behavior: equilibrium, periodicity, quasi-periodicity, deterministic chaos and random behavior. Systems adopt one or more these behaviors depending on the function they have evolved to perform. The emerging mathematical concepts of fractal mathematics and chaos theory are extending our ability to study physiological behavior. Fractal geometry is observed in the physical structure of pathways, networks and macroscopic structures such the vasculature and the His Purkinje network of the heart. Fractal structure is also observed in processes in time, such as heart rate variability. Chaos theory describes the underlying dynamics of the system, and chaotic behavior is also observed at many levels, from effector molecules in the cell to heart function and blood pressure. This review discusses the role of fractal structure and chaos in the cardiovascular system at the level of the heart and blood vessels, and at the cellular level. Key functional consequences of these phenomena are highlighted, and a perspective provided on the possible evolutionary origins of chaotic behavior and fractal structure. The discussion is non-mathematical with an emphasis on the key underlying concepts. PMID- 19812707 TI - Effects of clenbuterol, a beta2-adrenergic agonist, on sizes of masseter, temporalis, digastric, and tongue muscles. AB - We compared the hypertrophic effects of clenbuterol, a beta2-adrenergic agonist, on the masseter, digastric, and temporalis with those on the tongue, tibialis anterior, soleus, diaphragm, and heart. The weights of masseter, digastric and temporalis in the clenbuterol group were 36 ~ 56% greater than those in the control group, whereas those of the tibialis anterior, diaphragm, and heart weights in the clenbuterol group were 9 ~ 33% greater than those in the control group. No significant difference in the weights of the soleus and tongue was found between the control and clenbuterol groups. Taken together with our present and previously reported results, it is suggested that the hypertrophic effects of clenbuterol on the masseter, digastric, and temporalis are greater than those on the limb, trunk, and heart. PMID- 19812708 TI - Effects of cyclic strain and growth factors on vascular smooth muscle cell responses. AB - Under physiological and pathological conditions, vascular smooth muscle cells (SMC) are exposed to different biochemical factors and biomechanical forces. Previous studies pertaining to SMC responses have not investigated the effects of both factors on SMCs. Thus, in our research we investigated the combined effects of growth factors like Bfgf (basic fibroblast growth factor), TGF-beta (transforming growth factor beta) and PDGF (platelet-derived growth factor) along with physiological cyclic strain on SMC responses. Physiological cyclic strain (10% strain) significantly reduced SMC proliferation compared to static controls while addition of growth factors bFGF, TGF-beta or PDGF-AB had a positive influence on SMC growth compared to strain alone. Microarray analysis of SMCs exposed to these growth factors and cyclic strain showed that several bioactive genes (vascular endothelial growth factor, epidermal growth factor receptor, etc.) were altered upon exposure. Further work involving biochemical and pathological cyclic strain stimulation will help us better understand the role of cyclic strain and growth factors in vascular functions and development of vascular disorders. PMID- 19812709 TI - Adult congenital heart disease investigated with cardiac catheterization over a 20-year period. AB - BACKGROUND: Recent advances in diagnosis and treatment have increased the life expectancy of patients with congenital heart disease. METHODS: To investigate the prevalence of adult congenital heart disease (ACHD) in a large registry of patients over a 20-year period, we retrospectively assessed data of 14,012 males and 4,461 females who underwent clinically indicated cardiac catheterization from 1984 to 2003. RESULTS: ACHD was recorded in 234 subjects aged from 18 to 66 years, [95 males (40.7%) and 139 females (59.3%)]. Females were more likely to present with ACHD than males (p<0.001). Atrial septal defect was the most common defect (43.3%) followed by partial anomalous pulmonary venous return (12.0%), pulmonary valve stenosis (11.3%) ventricular septal defect (8.0%), coarctation of aorta (5.5%) patent ductus arteriosus (4.0%) and Fallot's tetralogy (3.3%). Atrial septal defect was more common in females (p<0.01), while pulmonary valve stenosis was more frequent in males (p<0.05). No difference across sexes was found in the other forms of ACHD. Females with ACHD were significantly older than males at the time of catheterization (median age 41 years, interquartile range 26 to 53 years vs. median age 35 years, interquartile range 22 to 48 years, p<0.05). CONCLUSIONS: In adulthood ACHD is found more commonly in females and is diagnosed later in life than in males. Atrial septal defect is the most prevalent form of ACHD and occurs most commonly in females. PMID- 19812710 TI - Addiction and free will. AB - Whether people believe that they have control over their behaviors is an issue that is centrally involved in definitions of addiction. Our research demonstrates that believing in free will - that is, believing that one has control over one's actions - has societal implications. Experimentally weakening free will beliefs led to cheating, stealing, aggression, and reduced helping. Bolstering free will beliefs did not change participants' behavior relative to a baseline condition, suggesting that most of the time people possess a belief in free will. We encourage a view of addiction that allows people to sustain a belief in free will and to take responsibility for choices and actions. PMID- 19812711 TI - Assessing inattention and impulsivity in children during the Go/NoGo task. AB - Behavioural performance in the Go/NoGo task was compared with caregiver and teacher reports of inattention and hyperactivity-impulsivity in 1,151 children (N = 557 boys; N = 594 girls) age 9- 10-years-old. Errors of commission (NoGo errors) were significantly correlated with symptom counts of hyperactivity impulsivity, while errors of omission (Go errors) were significantly correlated with symptom counts for inattention in both caregiver and teacher reports. Cross correlations were also evident, however, such that errors of commission were related to inattention symptoms, and errors of omission were related to hyperactivity-impulsivity. Moreover, hyperactivity-impulsivity and inattention symptoms were highly intercorrelated in both caregiver (r = .52) and teacher reports (r = .70), while errors of commission and omission were virtually uncorrelated in the Go/NoGo task (r = .06). The results highlight the difficulty in disentangling hyperactivity-impulsivity and inattention in questionnaires, and suggest that these constructs may be more clearly distinguished in laboratory measures such as the Go/NoGo task. PMID- 19812712 TI - Confocal Brillouin microscopy for three-dimensional mechanical imaging. AB - Acoustically induced inelastic light scattering, first reported in 1922 by Brillouin1, allows non-contact, direct readout of the viscoelastic properties of a material and has widely been investigated for material characterization2, structural monitoring3 and environmental sensing4. Extending the Brillouin technique from point sampling spectroscopy to imaging modality5 would open up new possibilities for mechanical imaging, but has been challenging because rapid spectrum acquisition is required. Here, we demonstrate a confocal Brillouin microscope based on a fully parallel spectrometer-a virtually imaged phased array that improves the detection efficiency by nearly 100-fold over previous approaches. Using the system, we show the first cross-sectional Brillouin imaging based on elastic properties as the contrast mechanism and monitor fast dynamic changes in elastic modulus during polymer crosslinking. Furthermore, we report the first in situ biomechanical measurement of the crystalline lens in a mouse eye. These results suggest multiple applications of Brillouin microscopy in biomedical and biomaterial science. PMID- 19812713 TI - A comparison of bounded diffusion models for choice in time controlled tasks. AB - The Wiener diffusion model (WDM) for 2-alternative tasks assumes that sensory information is integrated over time. Recent neurophysiological studies have found neural correlates of this integration process in certain neuronal populations. This paper analyses the properties of the WDM with two different boundary conditions in decision making tasks in which the time of response is indicated by a cue. A dual reflecting boundary mechanism is proposed and its performance is compared with a well-established absorbing boundary in the cases of the WDM, the WDM with extensions, and the WDM with prior probability. The two types of boundary influence the dynamics of the model and introduce differential weighting of evidence. Comparisons with Ornstein-Uhlenbeck models are also done, and it is shown that the WDM with both types of boundaries achieves similar performance and produce similar fits to existing behavioural data. Further studies are proposed to distinguish which boundary mechanism is more consistent with experimental data. PMID- 19812714 TI - Determining the effects of inositol supplementation and the opi1 mutation on ethanol tolerance of Saccharomyces cerevisiae. AB - The yeast Saccharomyces cerevisiae is an important microorganism for the ethanol fuel industry. As with many microorganisms, the production and accumulation of certain metabolites, such as ethanol, can have a detrimental effect on cell growth and productivity. Yeast cells containing a higher concentration of phosphatidylinositol (PI) in the cellular membrane, due to inositol supplementation in the growth media, have been shown to tolerate and produce higher concentrations of ethanol. The specific goal of our research was to assess the effects of inositol supplementation in the growth media as well as to compare the ethanol tolerance of the wild-type S. cerevisiae to a mutant, the opi1 strain (opi=overproduction of inositol). The OPI1 gene product is a negative regulatory factor that controls the transcription of the INO1 structural gene, which encodes the enzyme catalyzing the limiting step in the biosynthesis of inositol, that is, the conversion of glucose-6-phosphate to inositol-3-phosphate. Upon the deletion of the OPI1 gene, the cell will constitutively produce inositol, regardless of the extracellular inositol concentration. Inositol supplementation in cultures of wild-type cells increased ethanol tolerance in terms of cell viability. Cells grown in -I media had a 20% higher specific death rate than cells grown in +I media when exposed to 15% ethanol. The opi1 strain, with the ability to constitutively produce inositol regardless of media composition, showed less inhibition of cell growth in the presence of ethanol than did the wild-type strain, particularly in inositol-free media. We conclude that the introduction of an opi1 mutation in yeast results in an inherent increase in PI levels and constitutive biosynthesis of inositol that, in turn, will reduce the cost of supplementing inositol into the media to achieve a higher ethanol tolerance. PMID- 19812715 TI - Utilization of a biodegradable mulch sheet produced from poly(lactic acid)/ecoflex/modified starch in mandarin orange groves. AB - We have developed a mulch sheet made by inflation molding of PLA, Ecoflex((R)) and modified starch, which all have different biodegradabilities. A field test of use as an agricultural mulch sheet for mandarin oranges was carried out over two years. The mechanical properties of the mulch sheet were weakened with time during the field test, but the quality of the mandarin oranges increased, a result of the controlled degradation of the sheet. The most degradable modified starch degraded first, allowing control of the moisture on the soil. Accelerator mass spectroscopy was used for evaluation of the biomass carbon ratio. The biomass carbon ratio decreased by degradation of the biobased materials, PLA and modified starch in the mulch sheet. PMID- 19812716 TI - Diffusion in Model Networks as Studied by NMR and Fluorescence Correlation Spectroscopy. AB - We have studied the diffusion of small solvent molecules (octane) and larger hydrophobic dye probes in octane-swollen poly(dimethyl siloxane) linear-chain solutions and end-linked model networks, using pulsed-gradient nuclear magnetic resonance (NMR) and fluorescence correlation spectroscopy (FCS), respectively, focusing on diffusion in the bulk polymer up to the equilibrium degree of swelling of the networks, that is, 4.8 at most. The combination of these results allows for new conclusions on the feasibility of different theories describing probe diffusion in concentrated polymer systems. While octane diffusion shows no cross-link dependence, the larger dyes are increasingly restricted by fixed chemical meshes. The simple Fujita free-volume theory proved most feasible to describe probe diffusion in linear long-chain solutions with realistic parameters, while better fits were obtained assuming a stretched exponential dependence on concentration. Importantly, we have analyzed the cross-link specific effect on probe diffusion independently of any specific model by comparing the best-fit interpolation of the solution data with the diffusion in the networks. The most reasonable description is obtained by assuming that the cross-link effect is additive in the effective friction coefficient of the probes. The concentration dependences as well as the data compared at the equilibrium degrees of swelling indicate that swelling heterogeneities and diffusant shape have a substantial influence on small-molecule diffusion in networks. PMID- 19812717 TI - Intraocular Pressure: Does it Measure Up? AB - The relationship between intraocular pressure (IOP) and glaucoma is complex and not fully understood. We question the validity of several claims relating to the IOP-glaucoma relationship: (1) that 12 mm Hg is an important target in IOP control; (2) that IOP variability is an important risk factor for glaucoma progression; and (3) that every millimeter of mercury of IOP lowering reduces the risk of glaucoma progression by some specific percentage amount. Further, IOP is generally accepted to be an important - if not most important - risk factor for glaucoma development and progression. Using measures of treatment effect - absolute risk reduction, relative risk, and relative risk reduction - we compare the strength of IOP as a risk factor to the strength of a cardiac risk factor in cardiovascular disease. PMID- 19812718 TI - The influence of sex steroid hormones on gingiva of women. AB - Steroid sex hormones have a significant effect on different organ systems. As far as gingiva are concerned, they can influence the cellular proliferation, differentiation and growth of keratinocytes and fibroblasts. Estrogen is mainly responsible for alterations in blood vessels and progesterone stimulates the production of inflammatory mediators. In addition, some micro-organisms found in the human mouth synthesize enzymes needed for steroid synthesis and catabolism. In women, during puberty, ovulation and pregnancy, there is an increase in the production of sex steroid hormones which results in increased gingival inflammation, characterized by gingival enlargement, increased gingival bleeding and crevicular fluid flow and microbial changes. PMID- 19812719 TI - A target site for spontaneous insertion of IS10 element in pUC19 DNA located within intrinsically bent DNA. AB - Residual insertion sequence elements (IS elements) in Escherichia coli strains that are commonly used for DNA cloning are known to cause cloning artifacts by transposing themselves into the recombinant DNA fragments. In such cases, chance insertion of IS elements may occur at integration sites in the cloning targets, which in the case of the IS10 element is a 9-bp consensus sequence. We report here that the integration of IS10-related DNA sequences into the pUC19 cloning vector and its derivative occurred with considerable frequency in E. coli strains JM107 and DH10B, with duplication of a 9-bp segment (TCTAAAGTA). Notably, native polyacrylamide gel electrophoresis revealed that intrinsically bent DNA flanks the insertion site. PMID- 19812720 TI - Risk assessment for glaucoma. AB - Glaucoma is undergoing a paradigm shift and transitioning from merely disease staging to evidence-based risk assessment of in the individual patient.Initially introduced for ocular hypertensive patients, risk assessment calculators are now being developed for patients with established glaucoma. PMID- 19812721 TI - Activation of PyMT in beta cells induces irreversible hyperplasia, but oncogene dependent acinar cell carcinomas when activated in pancreatic progenitors. AB - It is unclear whether the cellular origin of various forms of pancreatic cancer involves transformation or transdifferentiation of different target cells or whether tumors arise from common precursors, with tumor types determined by the specific genetic alterations. Previous studies suggested that pancreatic ductal carcinomas might be induced by polyoma middle T antigen (PyMT) expressed in non ductal cells. To ask whether PyMT transforms and transdifferentiates endocrine cells toward exocrine tumor phenotypes, we generated transgenic mice that carry tetracycline-inducible PyMT and a linked luciferase reporter. Induction of PyMT in beta cells causes beta-cell hyperplastic lesions that do not progress to malignant neoplasms. When PyMT is de-induced, beta cell proliferation and growth cease; however, regression does not occur, suggesting that continued production of PyMT is not required to maintain the viable expanded beta cell population. In contrast, induction of PyMT in early pancreatic progenitor cells under the control of Pdx1 produces acinar cell carcinomas and beta-cell hyperplasia. The survival of acinar tumor cells is dependent on continued expression of PyMT. Our findings indicate that PyMT can induce exocrine tumors from pancreatic progenitor cells, but cells in the beta cell lineage are not transdifferentiated toward exocrine cell types by PyMT; instead, they undergo oncogene-dependent hyperplastic growth, but do not require PyMT for survival. PMID- 19812722 TI - REFGEN and TREENAMER: automated sequence data handling for phylogenetic analysis in the genomic era. AB - The phylogenetic analysis of nucleotide sequences and increasingly that of amino acid sequences is used to address a number of biological questions. Access to extensive datasets, including numerous genome projects, means that standard phylogenetic analyses can include many hundreds of sequences. Unfortunately, most phylogenetic analysis programs do not tolerate the sequence naming conventions of genome databases. Managing large numbers of sequences and standardizing sequence labels for use in phylogenetic analysis programs can be a time consuming and laborious task. Here we report the availability of an online resource for the management of gene sequences recovered from public access genome databases such as GenBank. These web utilities include the facility for renaming every sequence in a FASTA alignment file, with each sequence label derived from a user-defined combination of the species name and/or database accession number. This facility enables the user to keep track of the branching order of the sequences/taxa during multiple tree calculations and re-optimisations. Post phylogenetic analysis, these webpages can then be used to rename every label in the subsequent tree files (with a user-defined combination of species name and/or database accession number). Together these programs drastically reduce the time required for managing sequence alignments and labelling phylogenetic figures. Additional features of our platform include the automatic removal of identical accession numbers (recorded in the report file) and generation of species and accession number lists for use in supplementary materials or figure legends. PMID- 19812723 TI - Evolution and functional diversification of the GLI family of transcription factors in vertebrates. AB - BACKGROUND: In vertebrates the "SONIC HEDGEHOG" signalling pathway has been implicated in cell-fate determination, proliferation and the patterning of many different cell types and organs. As the GLI family members (GLI1, GLI2 and GLI3) are key mediators of hedgehog morphogenetic signals, over the past couple of decades they have been extensively scrutinized by genetic, molecular and biochemical means. Thus, a great deal of information is currently available about the functional aspects of GLI proteins in various vertebrate species. To address the roles of GLI genes in diversifying the repertoire of the Hh signalling and deploying them for the vertebrate specifications, in this study we have examined the evolutionary patterns of vertebrate GLI sequences within and between species. RESULTS: Phylogenetic tree analysis suggests that the vertebrate GLI1, GLI2 and GLI3 genes diverged after the separation of urochordates from vertebrates and before the tetrapods-bony fishes split. Lineage specific duplication events were also detected. Estimation of mode and strength of selection acting on GLI orthologs demonstrated that all members of the GLI gene family experienced more relaxed selection in teleost fish than in the mammalian lineage. Furthermore, the GLI1 gene appeared to have been exposed to different functional constraints in fish and tetrapod lineages, whilst a similar level of functional constraints on GLI2 and GLI3 was suggested by comparable average non-synonymous (Ka) substitutions across the lineages. A relative rate test suggested that the majority of the paralogous copies of the GLI family analyzed evolved with similar evolutionary rates except GLI1 which evolved at a significantly faster rate than its paralogous counterparts in tetrapods. CONCLUSIONS: Our analysis shows that sequence evolutionary patterns of GLI family members are largely correlated with the reported similarities and differences in the functionality of GLI proteins within and between the various vertebrate species. We propose that duplication and divergence of GLI genes has increased in the complexity of vertebrate body plan by recruiting the hedgehog signalling for the novel developmental tasks. PMID- 19812724 TI - Phylogenetic analyses reveal monophyletic origin of the ergot alkaloid gene dmaW in fungi. AB - Ergot alkaloids are indole-derived mycotoxins that are important in agriculture and medicine. Ergot alkaloids are produced by a few representatives of two distantly related fungal lineages, the Clavicipitaceae and the Trichocomaceae. Comparison of the ergot alkaloid gene clusters from these two lineages revealed differences in the relative positions and orientations of several genes. The question arose: is ergot alkaloid biosynthetic capability from a common origin? We used a molecular phylogenetic approach to gain insights into the evolution of ergot alkaloid biosynthesis. The 4-gamma,gamma-dimethylallyltryptophan synthase gene, dmaW, encodes the first step in the pathway. Amino acid sequences deduced from dmaW and homologs were submitted to phylogenetic analysis, and the results indicated that dmaW of Aspergillus fumigatus (mitosporic Trichocomaceae) has the same origin as corresponding genes from clavicipitaceous fungi. Relationships of authentic dmaW genes suggest that they originated from multiple gene duplications with subsequent losses of original or duplicate versions in some lineages. PMID- 19812725 TI - Phylogenetic applications of the minimum contradiction approach on continuous characters. AB - We describe the conditions under which a set of continuous variables or characters can be described as an X-tree or a split network. A distance matrix corresponds exactly to a split network or a valued X-tree if, after ordering of the taxa, the variables values can be embedded into a function with at most a local maximum and a local minimum, and crossing any horizontal line at most twice. In real applications, the order of the taxa best satisfying the above conditions can be obtained using the Minimum Contradiction method. This approach is applied to 2 sets of continuous characters. The first set corresponds to craniofacial landmarks in Hominids. The contradiction matrix is used to identify possible tree structures and some alternatives when they exist. We explain how to discover the main structuring characters in a tree. The second set consists of a sample of 100 galaxies. In that second example one shows how to discretize the continuous variables describing physical properties of the galaxies without disrupting the underlying tree structure. PMID- 19812726 TI - Initial implementation of a comparative data analysis ontology. AB - Comparative analysis is used throughout biology. When entities under comparison (e.g. proteins, genomes, species) are related by descent, evolutionary theory provides a framework that, in principle, allows N-ary comparisons of entities, while controlling for non-independence due to relatedness. Powerful software tools exist for specialized applications of this approach, yet it remains under utilized in the absence of a unifying informatics infrastructure. A key step in developing such an infrastructure is the definition of a formal ontology. The analysis of use cases and existing formalisms suggests that a significant component of evolutionary analysis involves a core problem of inferring a character history, relying on key concepts: "Operational Taxonomic Units" (OTUs), representing the entities to be compared; "character-state data" representing the observations compared among OTUs; "phylogenetic tree", representing the historical path of evolution among the entities; and "transitions", the inferred evolutionary changes in states of characters that account for observations. Using the Web Ontology Language (OWL), we have defined these and other fundamental concepts in a Comparative Data Analysis Ontology (CDAO). CDAO has been evaluated for its ability to represent token data sets and to support simple forms of reasoning. With further development, CDAO will provide a basis for tools (for semantic transformation, data retrieval, validation, integration, etc.) that make it easier for software developers and biomedical researchers to apply evolutionary methods of inference to diverse types of data, so as to integrate this powerful framework for reasoning into their research. PMID- 19812727 TI - A mixture model and a hidden markov model to simultaneously detect recombination breakpoints and reconstruct phylogenies. AB - Homologous recombination is a pervasive biological process that affects sequences in all living organisms and viruses. In the presence of recombination, the evolutionary history of an alignment of homologous sequences cannot be properly depicted by a single bifurcating tree: some sites have evolved along a specific phylogenetic tree, others have followed another path. Methods available to analyse recombination in sequences usually involve an analysis of the alignment through sliding-windows, or are particularly demanding in computational resources, and are often limited to nucleotide sequences. In this article, we propose and implement a Mixture Model on trees and a phylogenetic Hidden Markov Model to reveal recombination breakpoints while searching for the various evolutionary histories that are present in an alignment known to have undergone homologous recombination. These models are sufficiently efficient to be applied to dozens of sequences on a single desktop computer, and can handle equivalently nucleotide or protein sequences. We estimate their accuracy on simulated sequences and test them on real data. PMID- 19812728 TI - A comprehensive analysis of gene expression evolution between humans and mice. AB - Evolutionary changes in gene expression account for most phenotypic differences between species. Advances in microarray technology have made the systematic study of gene expression evolution possible. In this study, gene expression patterns were compared between human and mouse genomes using two published methods. Specifically, we studied how gene expression evolution was related to GO terms and tried to decode the relationship between promoter evolution and gene expression evolution. The results showed that (1) the significant enrichment of biological processes in orthologs of expression conservation reveals functional significance of gene expression conservation. The more conserved gene expression in some biological processes than is expected in a purely neutral model reveals negative selection on gene expression. However, fast evolving genes mainly support the neutrality of gene expression evolution, and (2) gene expression conservation is positively but only slightly correlated with promoter conservation based on a motif-count score of the promoter alignment. Our results suggest a neutral model with negative selection for gene expression evolution between humans and mice, and promoter evolution could have some effects on gene expression evolution. PMID- 19812729 TI - PhyLIS: a simple GNU/Linux distribution for phylogenetics and phyloinformatics. AB - PhyLIS is a free GNU/Linux distribution that is designed to provide a simple, standardized platform for phylogenetic and phyloinformatic analysis. The operating system incorporates most commonly used phylogenetic software, which has been pre-compiled and pre-configured, allowing for straightforward application of phylogenetic methods and development of phyloinformatic pipelines in a stable Linux environment. The software is distributed as a live CD and can be installed directly or run from the CD without making changes to the computer. PhyLIS is available for free at http://www.eve.ucdavis.edu/rcthomson/phylis/. PMID- 19812730 TI - On the use of bootstrapped topologies in coalescent-based Bayesian MCMC inference: a comparison of estimation and computational efficiencies. AB - Coalescent-based Bayesian Markov chain Monte Carlo (MCMC) inference generates estimates of evolutionary parameters and their posterior probability distributions. As the number of sequences increases, the length of time taken to complete an MCMC analysis increases as well. Here, we investigate an approach to distribute the MCMC analysis across a cluster of computers. To do this, we use bootstrapped topologies as fixed genealogies, perform a single MCMC analysis on each genealogy without topological rearrangements, and pool the results across all MCMC analyses. We show, through simulations, that although the standard MCMC performs better than the bootstrap-MCMC at estimating the effective population size (scaled by mutation rate), the bootstrap-MCMC returns better estimates of growth rates. Additionally, we find that our bootstrap-MCMC analyses are, on average, 37 times faster for equivalent effective sample sizes. PMID- 19812731 TI - A model for protein sequence evolution based on selective pressure for protein stability: application to hemoglobins. AB - Negative selection against protein instability is a central influence on evolution of proteins. Protein stability is maintained over evolution despite changes in underlying sequences. An empirical all-site stability-based model of evolution was developed to focus on the selection of residues arising from their contributions to protein stability. In this model, site rates could vary. A structure-based method was used to predict stationary frequencies of hemoglobin residues based on their propensity to promote protein stability at a site. Sites with destabilizing residues were shown to change more rapidly in hemoglobins than sites with stabilizing residues. For diverse proteins the results were consistent with stability-based selection. Maximum likelihood studies with hemoglobins supported the stability-based model over simple Poisson-based methods. These observations are consistent with suggestions that purifying selection to maintain protein structural stability plays a dominant role in protein evolution. PMID- 19812732 TI - Lead Optimization in Discovery Drug Metabolism and Pharmacokinetics/Case study: The Hepatitis C Virus (HCV) Protease Inhibitor SCH 503034. AB - Lead optimization using drug metabolism and pharmacokinetics (DMPK) parameters has become one of the primary focuses of research organizations involved in drug discovery in the last decade. Using a combination of rapid in vivo and in vitro DMPK screening procedures on a large array of compounds during the lead optimization process has resulted in development of compounds that have acceptable DMPK properties. In this review, we present a general screening paradigm that is currently being used as part of drug discovery at Schering Plough and we describe a case study using the Hepatitis C Virus (HCV) protease inhibitor program as an example. By using the DMPK optimization tools, a potent HCV protease inhibitor, SCH 503034, was selected for development as a candidate drug. PMID- 19812733 TI - FTY720 story. Its discovery and the following accelerated development of sphingosine 1-phosphate receptor agonists as immunomodulators based on reverse pharmacology. AB - Fingolimod (FTY720) is the first of a novel class: sphingosine 1-phosphate (S1P) receptor modulator and is currently in phase 3 clinical trials for multiple sclerosis (MS). FTY720 was first synthesized in 1992 by chemical modification of an immunosuppressive natural product, ISP-I (myriocin). ISP-I was isolated from the culture broth of Isaria sinclairii, a type of vegetative wasp that was an 'eternal youth' nostrum in traditional Chinese medicine. ISP-I is an amino acid having three successive asymmetric centers and some functionalities. We simplified the structure drastically to find a nonchiral symmetric 2-substitued-2 aminopropane-1,3-diol framework for an in vivo immunosuppressive activity (inhibition of rat skin allograft rejection test or prolonging effect on rat skin allograft survival) and finally discovered FTY720. During the course of the lead optimization process, we encountered an unexpected dramatic change of the mechanism of action with an in vivo output unchanged. Since it proved that FTY720 did not inhibit serine palmitoyltransferase that is the target enzyme of ISP-I, reverse pharmacological approaches have been preformed to elucidate that FTY720 is mainly phosphorylated by sphingosine kinease 2 in vivo and the phosphorylated drug acts as a potent agonist of four of the five G protein coupled receptors for S1P: S1P(1), S1P(3), S1P(4) and S1P(5). Evidence has accumulated that immunomodulation by FTY720-P is based on agonism at the S1P(1) receptor. Medicinal chemistry targeting S1P(1) receptor agonists is currently in progress. The FTY720 story provides a methodology where in vivo screens rather than in vitro screens play important roles in the lead optimization. Unlike recent drug discovery methodologies, such a strategy as adopted by the FTY720 program would more likely meet serendipity. PMID- 19812734 TI - The pK(a) Distribution of Drugs: Application to Drug Discovery. AB - The acid-base dissociation constant (pK(a)) of a drug is a key physicochemical parameter influencing many biopharmaceutical characteristics. While this has been well established, the overall proportion of non-ionizable and ionizable compounds for drug-like substances is not well known. Even less well known is the overall distribution of acid and base pK(a) values. The current study has reviewed the literature with regard to both the proportion of ionizable substances and pK(a) distributions. Further to this a set of 582 drugs with associated pK(a) data was thoroughly examined to provide a representative set of observations. This was further enhanced by delineating the compounds into CNS and non-CNS drugs to investigate where differences exist. Interestingly, the distribution of pK(a) values for single acids differed remarkably between CNS and non-CNS substances with only one CNS compound having an acid pK(a) below 6.1. The distribution of basic substances in the CNS set also showed a marked cut off with no compounds having a pK(a) above 10.5.The pK(a) distributions of drugs are influenced by two main drivers. The first is related to the nature and frequency of occurrence of the functional groups that are commonly observed in pharmaceuticals and the typical range of pK(a) values they span. The other factor concerns the biological targets these compounds are designed to hit. For example, many CNS targets are based on seven transmembrane G protein-coupled receptors (7TM GPCR) which have a key aspartic acid residue known to interact with most ligands. As a consequence, amines are mostly present in the ligands that target 7TM GPCR's and this influences the pK(a) profile of drugs containing basic groups. For larger screening collections of compounds, synthetic chemistry and the working practices of the chemists themselves can influence the proportion of ionizable compounds and consequent pK(a) distributions. The findings from this study expand on current wisdom in pK(a) research and have implications for discovery research with regard to the composition of corporate databases and collections of screening compounds. Rough guidelines have been suggested for the profile of compound collections and will evolve as this research area is expanded. PMID- 19812735 TI - Chemical chaperone and inhibitor discovery: potential treatments for protein conformational diseases. AB - Protein misfolding and aggregation cause a large number of neurodegenerative diseases in humans due to (i) gain of function as observed in Alzheimer's disease, Huntington's disease, Parkinson's disease, and Prion's disease or (ii) loss of function as observed in cystic fibrosis and alpha1-antitrypsin deficiency. These misfolded proteins could either lead to the formation of harmful amyloids that become toxic for the cells or to be recognized and prematurely degraded by the protein quality control system. An increasing number of studies has indicated that some low-molecular-weight compounds named as chemical chaperones can reverse the mislocalization and/or aggregation of proteins associated with human conformational diseases. These small molecules are thought to non-selectively stabilize proteins and facilitate their folding. In this review, we summarize the probable mechanisms of protein conformational diseases in humans and the use of chemical chaperones and inhibitors as potential therapeutic agents against these diseases. Furthermore, recent advanced experimental and theoretical approaches underlying the detailed mechanisms of protein conformational changes and current structure-based drug designs towards protein conformational diseases are also discussed. It is believed that a better understanding of the mechanisms of conformational changes as well as the biological functions of these proteins will lead to the development and design of potential interfering compounds against amyloid formation associated with protein conformational diseases. PMID- 19812736 TI - Derivatives of 5-aminolevulinic Acid for photodynamic therapy. AB - Photodynamic therapy (PDT) is a clinical treatment that combines the effects of visible light irradiation with subsequent biochemical events that arise from the presence of a photosensitising drug (possessing no dark toxicity) to cause destruction of selected cells. Today, the most common agent used in dermatological PDT is 5-aminolevulinic acid (ALA). As a result of its hydrophilic character, ALA penetrates skin lesions poorly when applied topically. Its systemic bioavailability is limited and it is known to cause significant side effects when given orally or intravenously. Numerous chemical derivatives of ALA have been synthesised with the aims of either improving topical penetration or enhancing systemic bioavailability, while reducing side effects. In vitro cell culture experiments with ALA derivatives have yielded promising results. However, if ALA derivatives are to demonstrate meaningful clinical benefits, a rational approach to topical formulation design is required, along with a systematic study aimed at uncovering the true potential of ALA derivatives in photodynamic therapy. With respect to systemic ALA delivery, more study is required in the developing area of ALA-containing dendrons and dendrimers. PMID- 19812738 TI - Perspectives on using physcomitrella patens as an alternative production platform for thapsigargin and other terpenoid drug candidates. AB - To overcome the potential future demand for terpenoids used as drugs, a new production platform is currently being established in our laboratory. The moss Physcomitrella has been chosen as the candidate organism for production of drug candidates based on terpenoids derived from plants, with a primary focus on the sesquiterpene lactone, thapsigargin. This drug candidate and other candidates/drugs with sesquiterpene skeleton are difficult to obtain by chemical synthesis due to their large number of chiral centers. Furthermore, they are not available in sufficient amounts from their original plant. The requirement for a new production system to meet the potential market demand for these compounds is not only obvious, but also essential if sufficient quantities of the drug candidates are to be available for the potential therapeutic use. PMID- 19812737 TI - The role of glucose metabolism and glucose-associated signalling in cancer. AB - Aggressive carcinomas ferment glucose to lactate even in the presence of oxygen. This particular metabolism, termed aerobic glycolysis, the glycolytic phenotype, or the Warburg effect, was discovered by Nobel laureate Otto Warburg in the 1920s. Since these times, controversial discussions about the relevance of the fermentation of glucose by tumours took place; however, a majority of cancer researchers considered the Warburg effect as a non-causative epiphenomenon. Recent research demonstrated, that several common oncogenic events favour the expression of the glycolytic phenotype. Moreover, a suppression of the phenotypic features by either substrate limitation, pharmacological intervention, or genetic manipulation was found to mediate potent tumour-suppressive effects. The discovery of the transketolase-like 1 (TKTL1) enzyme in aggressive cancers may deliver a missing link in the interpretation of the Warburg effect. TKTL1 activity could be the basis for a rapid fermentation of glucose in aggressive carcinoma cells via the pentose phosphate pathway, which leads to matrix acidification, invasive growth, and ultimately metastasis. TKTL1 expression in certain non-cancerous tissues correlates with aerobic formation of lactate and rapid fermentation of glucose, which may be required for the prevention of advanced glycation end products and the suppression of reactive oxygen species. There is evidence, that the activity of this enzyme and the Warburg effect can be both protective or destructive for the organism. These results place glucose metabolism to the centre of pathogenesis of several civilisation related diseases and raise concerns about the high glycaemic index of various food components commonly consumed in western diets. PMID- 19812739 TI - Chaperone therapy for neuronopathic lysosomal diseases: competitive inhibitors as chemical chaperones for enhancement of mutant enzyme activities. AB - Chaperone therapy is a newly developed molecular approach to lysosomal diseases, a group of human genetic diseases causing severe brain damage. We found two valienamine derivatives, N-octyl-4-epi-beta-valienamine (NOEV) and N-octyl-beta valienamine (NOV), as promising therapeutic agents for human beta-galactosidase deficiency disorders (mainly G(M1)-gangliosidosis) and beta-glucosidase deficiency disorders (Gaucher disease), respectively. We briefly reviewed the historical background of research in carbasugar glycosidase inhibitors. Originally NOEV and NOV had been discovered as competitive inhibitors, and then their paradoxical bioactivities as chaperones were confirmed in cultured fibroblasts from patients with these disorders. Subsequently G(M1)-gangliosidosis model mice were developed and useful for experimental studies. Orally administered NOEV entered the brain through the blood-brain barrier, enhanced beta-galactosidase activity, reduced substrate storage, and improved neurological deterioration clinically. Furthermore, we executed computational analysis for prediction of molecular interactions between beta-galactosidase and NOEV. Some preliminary results of computational analysis of molecular interaction mechanism are presented in this article. NOV also showed the chaperone effect toward several beta-glucosidase gene mutations in Gaucher disease. We hope chaperone therapy will become available for some patients with G(M1)-gangliosidosis, Gaucher disease, and potentially other lysosomal storage diseases with central nervous system involvement. PMID- 19812740 TI - Aminoglycosides: molecular insights on the recognition of RNA and aminoglycoside mimics. AB - RNA is increasingly recognized for its significant functions in biological systems and has recently become an important molecular target for therapeutics development. Aminoglycosides, a large class of clinically significant antibiotics, exert their biological functions by binding to prokaryotic ribosomal RNA (rRNA) and interfering with protein translation, resulting in bacterial cell death. They are also known to bind to viral mRNAs such as HIV-1 RRE and TAR. Consequently, aminoglycosides are accepted as the single most important model in understanding the principles that govern small molecule-RNA recognition, which is essential for the development of novel antibacterial, antiviral or even anti oncogenic agents. This review outlines the chemical structures and mechanisms of molecular recognition and antibacterial activity of aminoglycosides and various aminoglycoside mimics that have recently been devised to improve biological efficacy, binding affinity and selectivity, or to circumvent bacterial resistance. PMID- 19812741 TI - WPA-WHO collaborative activities 2009-2011. PMID- 19812742 TI - Culture, cultural factors and psychiatric diagnosis: review and projections. AB - This paper aims to provide conceptual justifications for the inclusion of culture and cultural factors in psychiatric diagnosis, and logistic suggestions as to the content and use of this approach. A discussion of the scope and limitations of current diagnostic practice, criticisms from different quarters, and the role and relevance of culture in the diagnostic encounter, precede the examination of advantages and disadvantages of the approach. The cultural content of psychiatric diagnosis should include the main, well-recognized cultural variables, adequate family data, explanatory models, and strengths and weaknesses of every individual patient. The practical aspects include the acceptance of "cultural discordances" as a component of an updated definition of mental disorder, and the use of a refurbished cultural formulation. Clinical "telescoping" strategies to obtain relevant cultural data during the diagnostic interview, and areas of future research (including field trials on the cultural formulation and on "culture bound syndromes"), are outlined. PMID- 19812744 TI - Salient components of a comprehensive service for eating disorders. AB - Eating disorders are challenging and difficult to treat, because of the necessity of a multidisciplinary treatment team for effective outcomes and the high mortality rate of anorexia nervosa. An adequate initial assessment and evaluation requires a psychiatric assessment, a medical history and medical examination, a social history and an interview of family members or collateral informants. A comprehensive eating disorder treatment team includes a psychiatrist coordinating the treatment and appropriate medical physician specialists, nutritionists, and psychotherapists. An adequate outpatient eating disorder clinic needs to provide individual psychotherapy with cognitive behavioral techniques specific for anorexia nervosa and bulimia nervosa, family therapy, pharmacological treatment and the resources to obtain appropriate laboratory tests. Eating disorder patients requiring inpatient care are best treated in a specialized eating disorder inpatient unit. A cognitive behavioral framework is most useful for the overall unit milieu. Medical management and nutritional rehabilitation are the primary goals for inpatient treatment. Various group therapies can cover common core eating disorder psychopathology problems and dialectical behavior therapy groups can be useful for managing emotional dysregulation. Residential, partial hospitalization and day treatment programs are useful for transitioning patients from an inpatient program or for patients needing some monitoring. In these programs, at least one structured meal is advisable as well as nutritional counseling, group therapy or individual counseling sessions. Group therapies usually address issues such as social skills training, social anxiety, body image distortion or maturity fears. Unfortunately there is s paucity of evidence based randomized control trials to recommend the salient components for a comprehensive service for eating disorders. Experienced eating disorder clinicians have come to the conclusion that a multidisciplinary team approach provides the most effective treatment. PMID- 19812745 TI - Diagnosis, provision and service organization. PMID- 19812743 TI - Research advances in geriatric depression. AB - Technical advances have facilitated the exploration of factors related to geriatric depression and have helped generate novel biological and psychosocial treatment approaches. This review summarizes the main advancements in epidemiology, clinical presentation and course, genetics, and other areas of biological research. Treatment interventions outlined in this paper include electroconvulsive therapy, repetitive transcranial magnetic stimulation, magnetic seizure therapy, vagus nerve stimulation, deep brain stimulatn, depression prophylaxis, multidisciplinary approaches to depression treatment, and psychotherapy. Forms of psychotherapy for geriatric depression summarized include interpersonal psychotherapy, supportive psychotherapy, cognitive-behavioral therapy, problem-solving therapy, and ecosystem-focused therapy. Neuroimaging techniques based on magnetic resonance imaging are discussed briefly, including volumetric brain studies, diffusion tensor imaging, fractional anisotropy, fiber tractography, magnetization transfer imaging, and blood-oxygenation-level dependent functional magnetic resonance imaging. Finally, treatment effectiveness is addressed in a discussion of new models to improve access to and quality of care offered in the community. PMID- 19812746 TI - Planning an eating disorder service on the basis of epidemiological data. PMID- 19812747 TI - A comprehensive treatment service must include developmental, systemic and collaborative components. PMID- 19812748 TI - Psychiatrists, milieu and glue. PMID- 19812749 TI - Services for eating disorders: how comprehensive is comprehensive? PMID- 19812750 TI - The eating disorders milieu. PMID- 19812751 TI - Therapeutic approach to eating disorders: the biological background. PMID- 19812752 TI - Mindfulness in the management of eating disorders. PMID- 19812753 TI - Management of eating disorders: optimal vs. minimum treatment. PMID- 19812754 TI - Continuum of depressive and manic mixed states in patients with bipolar disorder: quantitative measurement and clinical features. AB - Bipolar mixed states combine depressive and manic features, presenting diagnostic and treatment challenges and reflecting a severe form of the illness. DSM-IV criteria for a mixed state require combined depressive and manic syndromes, but a range of mixed states has been described clinically. A unified definition of mixed states would be valuable in understanding their diagnosis, mechanism and treatment implications. We investigated the manner in which depressive and manic features combine to produce a continuum of mixed states. In 88 subjects with bipolar disorder (DSM-IV), we evaluated symptoms and clinical characteristics, and compared depression-based, mania-based, and other published definitions of mixed states. We developed an index of the extent to which symptoms were mixed (Mixed State Index, MSI) and characterized its relationship to clinical state. Predominately manic and depressive mixed states using criteria from recent literature, as well as Kraepelinian mixed states, had similar symptoms and MSI scores. Anxiety correlated significantly with depression scores in manic subjects and with mania scores in depressed subjects. Discriminant function analysis associated mixed states with symptoms of hyperactivity and negative cognitions, but not subjective depressive or elevated mood. High MSI scores were associated with severe course of illness. For depressive or manic episodes, characteristics of mixed states emerged with two symptoms of the opposite polarity. This was a cross-sectional study. Mixed states appear to be a continuum. An index of the degree to which depressive and manic symptoms combine appears useful in identifying and characterizing mixed states. We propose a depressive or manic episode with three or more symptoms of the opposite polarity as a parsimonious definition of a mixed state. PMID- 19812755 TI - Factors predicting drop-out in community mental health centres. AB - This study aimed to identify treatment, therapist and patient factors associated with dropping out of treatment in four outpatient mental health services. The experimental group comprised all 789 individuals who attended for the first time the mental health services during one year and dropped out of treatment in the same year or during the two following ones. The control group consisted of the same number of individuals, chosen at random from patients who, in the same year, attended for the first time the services and did not subsequently drop out of treatment. The overall drop-out rate was 33.2%. According to logistic regression analysis, the predictive factors of dropping out were: being treated in a particular centre, the involvement of more than one therapist in treatment, having no previous history of psychiatric disorders, being young and being male. PMID- 19812756 TI - Somatic and cognitive domains of depression in an underserved region of Ecuador: some cultural considerations. AB - Not enough research efforts on depression have been carried out up to now in Latin America. The knowledge that has resulted from research activities in the United States or Europe offers limited generalizability to other regions of the world, including Latin America. In the Andean highlands of Ecuador, we found very high rates of moderate and severe depressive symptoms, a finding that must be interpreted within its cultural context. Somatic manifestations of depression predominated over cognitive manifestations, and higher education level was protective against depression. These findings call for an appreciation of culturally-specific manifestations of depression and the social factors that influence them. These factors must be further studied in order to give them the deserved priority, allocate resources appropriately, and formulate innovative psychosocial interventions. PMID- 19812757 TI - International guide to prescribing psychotropic medication for the management of problem behaviours in adults with intellectual disabilities. AB - Psychotropic medications are used regularly to manage problem behaviours among people with intellectual disabilities. This causes concern because often these medications are used out of their licensed indications in this context. The WPA Section on Psychiatry of Intellectual Disability has recently developed an evidence and consensus-based international guide for practitioners for the use of psychotropic medications for problem behaviours among adults with intellectual disabilities. This guide advises on assessment of behaviours, producing a formulation, initiation of treatment, assessment of out-come and adverse effects, follow-up arrangements, and possibility of discontinuation of treatment. PMID- 19812758 TI - Mental patients in prisons. AB - Mental conditions usually affect cognitive, emotional and volitional aspects and functions of the personality, which are also functions of interest in law, as they are essential at the time of adjudicating guilt, labeling the accused a criminal, and proffering a sentence. A relationship between mental illness and criminality has, thus, been described and given as one of the reasons for the large number of mental patients in prisons. Whether this relationship is one of causality or one that flows through many other variables is a matter of debate, but there is no debating that prisons have become a de facto part, and an important one, of mental health systems in many countries. This paper deals with the issue of the relationship and provides estimates of prevalence of mental patients in prisons culled from many studies in different countries. It also provides some direction for the management of mental patients as they crowd correctional systems. PMID- 19812759 TI - The WPA train-the-trainers workshop on mental health in primary care (Ibadan, Nigeria, January 26-30, 2009). PMID- 19812760 TI - Update on WPA education programs, 2009. PMID- 19812761 TI - WPA forthcoming scientific meetings. PMID- 19812762 TI - Proteomic analysis in diabetic cardiomyopathy using bioinformatics approach. AB - Diabetic cardiomyopathy is a distinct clinical entity that produces asymptomatic heart failure in diabetic patients without evidence of coronary artery disease and hypertension. Abnormalities in diabetic cardiomyopathy include: myocardial hypertrophy, impairment of contractile proteins, accumulation of extracellular matrix proteins, formation of advanced glycation end products, and decreased left ventricular compliance. These abnormalities lead to the most common clinical presentation of diabetic cardiomyopathy in the form of diastolic dysfunction.We evaluated the role of various proteins that are likely to be involved in diabetic cardiomyopathy by employing multiple sequence alignment using ClustalW tool and constructed a Phylogenetic tree using functional protein sequences extracted from NCBI. Phylogenetic tree was constructed using Neighbour-Joining Algorithm in bioinformatics approach. These results suggest a causal relationship between altered calcium homeostasis and diabetic cardiomyopathy that implies that efforts directed to normalize calcium homeostasis could form a novel therapeutic approach. PMID- 19812763 TI - Structural re-alignment in an immunogenic surface region of ricin A chain. AB - We compared structure alignments generated by several protein structure comparison programs to determine whether existing methods would satisfactorily align residues at a highly conserved position within an immunogenic loop in ribosome inactivating proteins (RIPs). Using default settings, structure alignments generated by several programs (CE, DaliLite, FATCAT, LGA, MAMMOTH, MATRAS, SHEBA, SSM) failed to align the respective conserved residues, although LGA reported correct residue-residue (R-R) correspondences when the beta-carbon (Cb) position was used as the point of reference in the alignment calculations. Further tests using variable points of reference indicated that points distal from the beta carbon along a vector connecting the alpha and beta carbons yielded rigid structural alignments in which residues known to be highly conserved in RIPs were reported as corresponding residues in structural comparisons between ricin A chain, abrin-A, and other RIPs. Results suggest that approaches to structure alignment employing alternate point representations corresponding to side chain position may yield structure alignments that are more consistent with observed conservation of functional surface residues than do standard alignment programs, which apply uniform criteria for alignment (i.e. alpha carbon (Ca) as point of reference) along the entirety of the peptide chain. We present the results of tests that suggest the utility of allowing user-specified points of reference in generating alternate structural alignments, and we present a web server for automatically generating such alignments: http://as2ts.llnl.gov/AS2TS/LGA/lga_pdblist_plots.html. PMID- 19812764 TI - Coarse-grained models reveal functional dynamics--I. Elastic network models- theories, comparisons and perspectives. AB - In this review, we summarize the progress on coarse-grained elastic network models (CG-ENMs) in the past decade. Theories were formulated to allow study of conformational dynamics in time/space frames of biological interest. Several highlighted models and their underlined hypotheses are introduced in physical depth. Important ENM offshoots, motivated to reproduce experimental data as well as to address the slow-mode-encoded configurational transitions, are also introduced. With the theoretical developments, computational cost is significantly reduced due to simplified potentials and coarse-grained schemes. Accumulating wealth of data suggest that ENMs agree equally well with experiment in describing equilibrium dynamics despite their distinct potentials and levels of coarse-graining. They however do differ in the slowest motional components that are essential to address large conformational changes of functional significance. The difference stems from the dissimilar curvatures of the harmonic energy wells described for each model. We also provide our views on the predictability of 'open to close' (open-->close) transitions of biomolecules on the basis of conformational selection theory. Lastly, we address the limitations of the ENM formalism which are partially alleviated by the complementary CG-MD approach, to be introduced in the second paper of this two-part series. PMID- 19812765 TI - Characterization of non-trivial neighborhood fold constraints from protein sequences using generalized topohydrophobicity. AB - Prediction of key features of protein structures, such as secondary structure, solvent accessibility and number of contacts between residues, provides useful structural constraints for comparative modeling, fold recognition, ab-initio fold prediction and detection of remote relationships. In this study, we aim at characterizing the number of non-trivial close neighbors, or long-range contacts of a residue, as a function of its "topohydrophobic" index deduced from multiple sequence alignments and of the secondary structure in which it is embedded. The "topohydrophobic" index is calculated using a two-class distribution of amino acids, based on their mean atom depths. From a large set of structural alignments processed from the FSSP database, we selected 1485 structural sub-families including at least 8 members, with accurate alignments and limited redundancy. We show that residues within helices, even when deeply buried, have few non-trivial neighbors (0-2), whereas beta-strand residues clearly exhibit a multimodal behavior, dominated by the local geometry of the tetrahedron (3 non-trivial close neighbors associated with one tetrahedron; 6 with two tetrahedra). This observed behavior allows the distinction, from sequence profiles, between edge and central beta-strands within beta-sheets. Useful topological constraints on the immediate neighborhood of an amino acid, but also on its correlated solvent accessibility, can thus be derived using this approach, from the simple knowledge of multiple sequence alignments. PMID- 19812766 TI - HMM_RA: an improved method for alpha-helical transmembrane protein topology prediction. AB - alpha-helical transmembrane (TM) proteins play important and diverse functional roles in cells. The ability to predict the topology of these proteins is important for identifying functional sites and inferring function of membrane proteins. This paper presents a Hidden Markov Model (referred to as HMM_RA) that can predict the topology of alpha-helical transmembrane proteins with improved performance. HMM_RA adopts the same structure as the HMMTOP method, which has five modules: inside loop, inside helix tail, membrane helix, outside helix tail and outside loop. Each module consists of one or multiple states. HMM_RA allows using reduced alphabets to encode protein sequences. Thus, each state of HMM_RA is associated with n emission probabilities, where n is the size of the reduced alphabet set. Direct comparisons using two standard data sets show that HMM_RA consistently outperforms HMMTOP and TMHMM in topology prediction. Specifically, on a high-quality data set of 83 proteins, HMM_RA outperforms HMMTOP by up to 7.6% in topology accuracy and 6.4% in alpha-helices location accuracy. On the same data set, HMM_RA outperforms TMHMM by up to 6.4% in topology accuracy and 2.9% in location accuracy. Comparison also shows that HMM_RA achieves comparable performance as Phobius, a recently published method. PMID- 19812767 TI - Estimating the fraction of non-coding RNAs in mammalian transcriptomes. AB - Recent studies of mammalian transcriptomes have identified numerous RNA transcripts that do not code for proteins; their identity, however, is largely unknown. Here we explore an approach based on sequence randomness patterns to discern different RNA classes. The relative z-score we use helps identify the known ncRNA class from the genome, intergene and intron classes. This leads us to a fractional ncRNA measure of putative ncRNA datasets which we model as a mixture of genuine ncRNAs and other transcripts derived from genomic, intergenic and intronic sequences. We use this model to analyze six representative datasets identified by the FANTOM3 project and two computational approaches based on comparative analysis (RNAz and EvoFold). Our analysis suggests fewer ncRNAs than estimated by DNA sequencing and comparative analysis, but the verity of our approach and its prediction requires more extensive experimental RNA data. PMID- 19812768 TI - Effects of feeding periods of high cholesterol and saturated fat diet on blood biochemistry and hydroxyproline fractions in rabbits. AB - Hypercholesterolemia and hypertriglyceridemia are considered as important risk factors during the atherosclerotic process. The aim of the present investigation was to study the total cholesterol (TC), low-density lipoprotein cholesterol (LDLC), high density lipoprotein (HDL), triglyceride (TG), platelet levels and hydroxyproline fractions during the pathogenesis of atherosclerosis. For this purpose, twenty five 12-weeks, New Zealand white male rabbits, were purchased, individually caged, and divided into either control group or cholesterol-fed group. The control group (n = 10) was fed 100 g/day of normal diet, ORC-4 (Oriental Yeast Co. Ltd., Tokyo, Japan) for a period of 15 weeks. The cholesterol fed group (n = 15) was fed a high cholesterol and saturated fat diet of ORC-4 containing 1% cholesterol plus 1% olive oil (100 g/day) for periods of 5 (group 1), 10 (group 2) and 15 (group 3) weeks. Blood sample from each animal was taken at the end of the experimental period for the biochemical analysis. The results of the present study showed that TC, LDLC, TG, HDLC and platelets were significantly (P < 0.01) increased in cholesterol-fed rabbits as compared with control rabbits. The serum hydroxyproline (Hyp) in rabbits belonging to group 1 showed no significant alteration when compared to control group. Group 2 rabbits showed a significant increase of 103% (P < 0.01) and 100% (P < 0.001) in free and protein-bound hydroxyproline fractions respectively when compared to control rabbits. However, there was no significant change in peptide-bound and total serum hydroxyproline levels as compared to the control group (P > 0.05). There was no significant (P > 0.05) decrease of free serum hydroxyproline in group 3 rabbits when compared to control rabbits. On the other hand, group 3 rabbits showed a significant increase in peptide-bound and protein-bound Hyp by 517% (P < 0.05) and 100% (P < 0.01) respectively when compared to control rabbits. However, total serum Hyp in group 3 rabbits showed no significant (P > 0.05) change when compared to control rabbits. These results suggest that feeding rabbits high cholesterol and saturated fat diet for feeding periods of 5, 10 and 15 weeks induced significant change in TC, LDLC, HDL, TG, platelet levels and various Hyp fractions in serum without any significant change in the total Hyp content. PMID- 19812769 TI - Utility of computational methods to identify the apoptosis machinery in unicellular eukaryotes. AB - Apoptosis is the phenotypic result of an active, regulated process of self destruction. Following various cellular insults, apoptosis has been demonstrated in numerous unicellular eukaryotes, but very little is known about the genes and proteins that initiate and execute this process in this group of organisms. A bioinformatic approach presents an array of powerful methods to direct investigators in the identification of the apoptosis machinery in protozoans. In this review, we discuss some of the available computational methods and illustrate how they may be applied using the identification of a Plasmodium falciparum metacaspase gene as an example. PMID- 19812770 TI - A unified discussion on the concept of score functions used in the context of nonparametric linkage analysis. AB - In this article we try to discuss nonparametric linkage (NPL) score functions within a broad and quite general framework. The main focus of the paper is the structure, derivation principles and interpretations of the score function entity itself. We define and discuss several families of one-locus score function definitions, i.e. the implicit, explicit and optimal ones. Some generalizations and comments to the two-locus, unconditional and conditional, cases are included as well. Although this article mainly aims at serving as an overview, where the concept of score functions are put into a covering context, we generalize the noncentrality parameter (NCP) optimal score functions in Angquist et al. (2007) to facilitate--through weighting--for incorporation of several plausible distinct genetic models. Since the genetic model itself most oftenly is to some extent unknown this facilitates weaker prior assumptions with respect to plausible true disease models without loosing the property of NCP-optimality.Moreover, we discuss general assumptions and properties of score functions in the above sense. For instance, the concept of identical by descent (IBD) sharing structures and score function equivalence are discussed in some detail. PMID- 19812771 TI - Searching for factors that distinguish disease-prone and disease-resistant prions via sequence analysis. AB - The exact mechanisms of prion misfolding and factors that predispose an individual to prion diseases are largely unknown. Our approach to identifying candidate factors in-silico relies on contrasting the C-terminal domain of PrP(C) sequences from two groups of vertebrate species: those that have been found to suffer from prion diseases, and those that have not. We propose that any significant differences between the two groups are candidate factors that may predispose individuals to develop prion disease, which should be further analyzed by wet-lab investigations. Using an array of computational methods we identified possible point mutations that could predispose PrP(C) to misfold into PrP(Sc). Our results include confirmatory findings such as the V210I mutation, and new findings including P137M, G142D, G142N, D144P, K185T, V189I, H187Y and T191P mutations, which could impact structural stability. We also propose new hypotheses that give insights into the stability of helix-2 and -3. These include destabilizing effects of Histidine and T188-T193 segment in helix-2 in the disease-prone prions, and a stabilizing effect of Leucine on helix-3 in the disease-resistant prions. PMID- 19812772 TI - A new test statistic based on shrunken sample variance for identifying differentially expressed genes in small microarray experiments. AB - Choosing an appropriate statistic and precisely evaluating the false discovery rate (FDR) are both essential for devising an effective method for identifying differentially expressed genes in microarray data. The t-type score proposed by Pan et al. (2003) succeeded in suppressing false positives by controlling the underestimation of variance but left the overestimation uncontrolled. For controlling the overestimation, we devised a new test statistic (variance stabilized t-type score) by placing shrunken sample variances of the James-Stein type in the denominator of the t-type score. Since the relative superiority of the mean and median FDRs was unclear in the widely adopted Significance Analysis of Microarrays (SAM), we conducted simulation studies to examine the performance of the variance stabilized t-type score and the characteristics of the two FDRs. The variance stabilized t-type score was generally better than or at least as good as the t-type score, irrespective of the sample size and proportion of differentially expressed genes. In terms of accuracy, the median FDR was superior to the mean FDR when the proportion of differentially expressed genes was large. The variance stabilized t-type score with the median FDR was applied to actual colorectal cancer data and yielded a reasonable result. PMID- 19812773 TI - Distribution of polymorphic and non-polymorphic microsatellite repeats in Xenopus tropicalis. AB - The results of our bioinformatics analysis have found over 91,000 di-, tri-, and tetranucleotide microsatellites in our survey of 25% of the X. tropicalis genome, suggesting there may be over 360,000 within the entire genome. Within the X. tropicalis genome, dinucleotide (78.7%) microsatellites vastly out numbered tri- and tetranucleotide microsatellites. Similarly, AT-rich repeats are overwhelmingly dominant. The four AT-only motifs (AT, AAT, AAAT, and AATT) account for 51,858 out of 91,304 microsatellites found. Individually, AT microsatellites were the most common repeat found, representing over half of all di-, tri-, and tetranucleotide microsatellites. This contrasts with data from other studies, which show that AC is the most frequent microsatellite in vertebrate genomes (Toth et al. 2000). In addition, we have determined the rate of polymorphism for 5,128 non-redundant microsatellites, embedded in unique sequences. Interestingly, this subgroup of microsatellites was determined to have significantly longer repeats than genomic microsatellites as a whole. In addition, microsatellite loci with tandem repeat lengths more than 30 bp exhibited a significantly higher degree of polymorphism than other loci. Pairwise comparisons show that tetranucleotide microsatellites have the highest polymorphic rates. In addition, AAT and ATC showed significant higher polymorphism than other trinucleotide microsatellites, while AGAT and AAAG were significantly more polymorphic than other tetranucleotide microsatellites. PMID- 19812774 TI - Coarse-grained models reveal functional dynamics--II. Molecular dynamics simulation at the coarse-grained level--theories and biological applications. AB - Molecular dynamics (MD) simulation has remained the most indispensable tool in studying equilibrium/non-equilibrium conformational dynamics since its advent 30 years ago. With advances in spectroscopy accompanying solved biocomplexes in growing sizes, sampling their dynamics that occur at biologically interesting spatial/temporal scales becomes computationally intractable; this motivated the use of coarse-grained (CG) approaches. CG-MD models are used to study folding and conformational transitions in reduced resolution and can employ enlarged time steps due to the absence of some of the fastest motions in the system. The Boltzmann-Inversion technique, heavily used in parameterizing these models, provides a smoothed-out effective potential on which molecular conformation evolves at a faster pace thus stretching simulations into tens of microseconds. As a result, a complete catalytic cycle of HIV-1 protease or the assembly of lipid-protein mixtures could be investigated by CG-MD to gain biological insights. In this review, we survey the theories developed in recent years, which are categorized into Folding-based and Molecular-Mechanics-based. In addition, physical bases in the selection of CG beads/time-step, the choice of effective potentials, representation of solvent, and restoration of molecular representations back to their atomic details are systematically discussed. PMID- 19812775 TI - Ontologies for bioinformatics. AB - The past twenty years have witnessed an explosion of biological data in diverse database formats governed by heterogeneous infrastructures. Not only are semantics (attribute terms) different in meaning across databases, but their organization varies widely. Ontologies are a concept imported from computing science to describe different conceptual frameworks that guide the collection, organization and publication of biological data. An ontology is similar to a paradigm but has very strict implications for formatting and meaning in a computational context. The use of ontologies is a means of communicating and resolving semantic and organizational differences between biological databases in order to enhance their integration. The purpose of interoperability (or sharing between divergent storage and semantic protocols) is to allow scientists from around the world to share and communicate with each other. This paper describes the rapid accumulation of biological data, its various organizational structures, and the role that ontologies play in interoperability. PMID- 19812776 TI - Borges dilemma, fundamental laws, and systems biology. PMID- 19812777 TI - Topological properties of co-occurrence networks in published gene expression signatures. AB - Meta-analysis of high-throughput gene expression data is often used for the interpretation of proprietary gene expression data sets. We have recently shown that co-occurrence patterns of gene expression in published cancer-related gene expression signatures are reminiscent of several cancer signaling pathways. Indeed, significant co-occurrence of up to ten genes in published gene expression signatures can be exploited to build a co-occurrence network from the sets of co occurring genes ("co-occurrence modules"). Such co-occurrence network is represented by an undirected graph, where single genes are assigned to vertices and edges indicate that two genes are significantly co-occurring. Thus, graph-cut methods can be used to identify groups of highly interconnected vertices ("network communities") that correspond to sets of genes that are significantly co-regulated in human cancer. Here, we investigate the topological properties of co-occurrence networks derived from published gene expression signatures and show that co-occurrence networks are characterized by scale-free topology and hierarchical modularity. Furthermore, we report that genes with a "promiscuous" or a "faithful" co-occurrence pattern can be distinguished. This behavior is reminiscent of date and party hubs that have been identified in protein-protein interaction networks. PMID- 19812778 TI - Evaluation of combining several statistical methods with a flexible cutoff for identifying differentially expressed genes in pairwise comparison of EST sets. AB - The detection of differentially expressed genes from EST data is of importance for the discovery of potential biological or pharmaceutical targets, especially when studying biological processes in less characterized organisms and where large-scale microarrays are not an option. We present a comparison of five different statistical methods for identifying up-regulated genes through pairwise comparison of EST sets, where one of the sets is generated from a treatment and the other one serves as a control. In addition, we specifically address situations where the sets are relatively small (micro 2,000-10,000 ESTs) and may differ in size. The methods were tested on both simulated and experimentally derived data, and compared to a collection of cold stress induced genes identified by microarrays. We found that combining the method proposed by Audic and Claverie with Fisher's exact test and a method based on calculating the difference in relative frequency was the best combination for maximizing the detection of up-regulated genes. We also introduced the use of a flexible cutoff, which takes the size of the EST sets into consideration. This could be considered as an alternative to a static cutoff. Finally, the detected genes showed a low overlap with those identified by microarrays, which indicates, as in previous studies, low overall concordance between the two platforms. PMID- 19812779 TI - Methodology for constructing problem definitions in bioinformatics. AB - MOTIVATION: A recurrent criticism is that certain bioinformatics tools do not account for crucial biology and therefore fail answering the targeted biological question. We posit that the single most important reason for such shortcomings is an inaccurate formulation of the computational problem. RESULTS: Our paper describes how to define a bioinformatics problem so that it captures both the underlying biology and the computational constraints for a particular problem. The proposed model delineates comprehensively the biological problem and conducts an item-by-item bioinformatics transformation resulting in a germane computational problem. This methodology not only facilitates interdisciplinary information flow but also accommodates emerging knowledge and technologies. PMID- 19812780 TI - Neuroinformatics: from bioinformatics to databasing the brain. AB - Neuroinformatics seeks to create and maintain web-accessible databases of experimental and computational data, together with innovative software tools, essential for understanding the nervous system in its normal function and in neurological disorders. Neuroinformatics includes traditional bioinformatics of gene and protein sequences in the brain; atlases of brain anatomy and localization of genes and proteins; imaging of brain cells; brain imaging by positron emission tomography (PET), functional magnetic resonance imaging (fMRI), electroencephalography (EEG), magnetoencephalography (MEG) and other methods; many electrophysiological recording methods; and clinical neurological data, among others. Building neuroinformatics databases and tools presents difficult challenges because they span a wide range of spatial scales and types of data stored and analyzed. Traditional bioinformatics, by comparison, focuses primarily on genomic and proteomic data (which of course also presents difficult challenges). Much of bioinformatics analysis focus on sequences (DNA, RNA, and protein molecules), as the type of data that are stored, compared, and sometimes modeled. Bioinformatics is undergoing explosive growth with the addition, for example, of databases that catalog interactions between proteins, of databases that track the evolution of genes, and of systems biology databases which contain models of all aspects of organisms. This commentary briefly reviews neuroinformatics with clarification of its relationship to traditional and modern bioinformatics. PMID- 19812781 TI - Unsupervised meta-analysis on diverse gene expression datasets allows insight into gene function and regulation. AB - Over the past years, microarray databases have increased rapidly in size. While they offer a wealth of data, it remains challenging to integrate data arising from different studies. Here we propose an unsupervised approach of a large-scale meta-analysis on Arabidopsis thaliana whole genome expression datasets to gain additional insights into the function and regulation of genes. Applying kernel principal component analysis and hierarchical clustering, we found three major groups of experimental contrasts sharing a common biological trait. Genes associated to two of these clusters are known to play an important role in indole 3-acetic acid (IAA) mediated plant growth and development or pathogen defense. Novel functions could be assigned to genes including a cluster of serine/threonine kinases that carry two uncharacterized domains (DUF26) in their receptor part implicated in host defense. With the approach shown here, hidden interrelations between genes regulated under different conditions can be unraveled. PMID- 19812782 TI - The use of artificial neural networks in prediction of congenital CMV outcome from sequence data. AB - A large number of CMV strains has been reported to circulate in the human population, and the biological significance of these strains is currently an active area of research. The analysis of complex genetic information may be limited using conventional phylogenetic techniques. We constructed artificial neural networks to determine their feasibility in predicting the outcome of congenital CMV disease (defined as presence of CMV symptoms at birth) based on two data sets: 54 sequences of CMV gene UL144 obtained from 54 amniotic fluids of women who contracted acute CMV infection during their pregnancy, and 80 sequences of 4 genes (US28, UL144, UL146 and UL147) obtained from urine, saliva or blood of 20 congenitally infected infants that displayed different outcomes at birth. When data from all four genes was used in the 20-infants' set, the artificial neural network model accurately identified outcome in 90% of cases. While US28 and UL147 had low yield in predicting outcome, UL144 and UL146 predicted outcome in 80% and 85% respectively when used separately. The model identified specific nucleotide positions that were highly relevant to prediction of outcome. The artificial neural network classified genotypes in agreement with classic phylogenetic analysis. We suggest that artificial neural networks can accurately and efficiently analyze sequences obtained from larger cohorts to determine specific outcomes.?The ANN training and analysis code is commercially available from Optimal Neural Informatics (Pikesville, MD). PMID- 19812783 TI - Normalization and gene p-value estimation: issues in microarray data processing. AB - INTRODUCTION: Numerous methods exist for basic processing, e.g. normalization, of microarray gene expression data. These methods have an important effect on the final analysis outcome. Therefore, it is crucial to select methods appropriate for a given dataset in order to assure the validity and reliability of expression data analysis. Furthermore, biological interpretation requires expression values for genes, which are often represented by several spots or probe sets on a microarray. How to best integrate spot/probe set values into gene values has so far been a somewhat neglected problem. RESULTS: We present a case study comparing different between-array normalization methods with respect to the identification of differentially expressed genes. Our results show that it is feasible and necessary to use prior knowledge on gene expression measurements to select an adequate normalization method for the given data. Furthermore, we provide evidence that combining spot/probe set p-values into gene p-values for detecting differentially expressed genes has advantages compared to combining expression values for spots/probe sets into gene expression values. The comparison of different methods suggests to use Stouffer's method for this purpose. The study has been conducted on gene expression experiments investigating human joint cartilage samples of osteoarthritis related groups: a cDNA microarray (83 samples, four groups) and an Affymetrix (26 samples, two groups) data set. CONCLUSION: The apparently straight forward steps of gene expression data analysis, e.g. between-array normalization and detection of differentially regulated genes, can be accomplished by numerous different methods. We analyzed multiple methods and the possible effects and thereby demonstrate the importance of the single decisions taken during data processing. We give guidelines for evaluating normalization outcomes. An overview of these effects via appropriate measures and plots compared to prior knowledge is essential for the biological interpretation of gene expression measurements. PMID- 19812784 TI - Evidence of highly regulated genes (in-Hubs) in gene networks of Saccharomyces cerevisiae. AB - Uncovering interactions between genes, gene networks, is important to increase our understanding of intrinsic cellular processes and responses to external stimuli such as drugs. Gene networks can be computationally inferred from repeated measurements of gene expression, using algorithms, which assume that each gene is controlled by only a small number of other proteins. Here, by extending the transcription network with cofactors (defined from protein-protein binding data) as active regulators, we identified the effective gene network, providing evidence of in-hubs in the gene regulatory networks of yeast. Then, using the notion that in-hub genes will be differentially expressed over several experimental conditions, we designed an algorithm, the HubDetector, enabling identification of in-hubs directly from gene expression data. Applying the HubDetector to 488 genome-wide expression profiles from two independent datasets, we identified putative in-hubs overlapping significantly with in-hubs in the effective gene network. PMID- 19812785 TI - Designing toxicogenomics studies that use DNA array technology. AB - BACKGROUND: Bioassays are routinely used to evaluate the toxicity of test agents. Experimental designs for bioassays are largely encompassed by fixed effects linear models. In toxicogenomics studies where DNA arrays measure mRNA levels, the tissue samples are typically generated in a bioassay. These measurements introduce additional sources of variation, which must be properly managed to obtain valid tests of treatment effects. RESULTS: An analysis of covariance model is developed which combines a fixed-effects linear model for the bioassay with important variance components associated with DNA array measurements. These models can accommodate the dominant characteristics of measurements from DNA arrays, and they account for technical variation associated with normalization, spots, dyes, and batches as well as the biological variation associated with the bioassay. An example illustrates how the model is used to identify valid designs and to compare competing designs. CONCLUSIONS: Many toxicogenomics studies are bioassays which measure gene expression using DNA arrays. These studies can be designed and analyzed using standard methods with a few modifications to account for characteristics of array measurements, such as multiple endpoints and normalization. As much as possible, technical variation associated with probes, dyes, and batches are managed by blocking treatments within these sources of variation. An example shows how some practical constraints can be accommodated by this modelling and how it allows one to objectively compare competing designs. PMID- 19812786 TI - Association between a prognostic gene signature and functional gene sets. AB - BACKGROUND: The development of expression-based gene signatures for predicting prognosis or class membership is a popular and challenging task. Besides their stringent validation, signatures need a functional interpretation and must be placed in a biological context. Popular tools such as Gene Set Enrichment have drawbacks because they are restricted to annotated genes and are unable to capture the information hidden in the signature's non-annotated genes. METHODOLOGY: We propose concepts to relate a signature with functional gene sets like pathways or Gene Ontology categories. The connection between single signature genes and a specific pathway is explored by hierarchical variable selection and gene association networks. The risk score derived from an individual patient's signature is related to expression patterns of pathways and Gene Ontology categories. Global tests are useful for these tasks, and they adjust for other factors. GlobalAncova is used to explore the effect on gene expression in specific functional groups from the interaction of the score and selected mutations in the patient's genome. RESULTS: We apply the proposed methods to an expression data set and a corresponding gene signature for predicting survival in Acute Myeloid Leukemia (AML). The example demonstrates strong relations between the signature and cancer-related pathways. The signature based risk score was found to be associated with development-related biological processes. CONCLUSIONS: Many authors interpret the functional aspects of a gene signature by linking signature genes to pathways or relevant functional gene groups. The method of gene set enrichment is preferred to annotating signature genes to specific Gene Ontology categories. The strategies proposed in this paper go beyond the restriction of annotation and deepen the insights into the biological mechanisms reflected in the information given by a signature. PMID- 19812787 TI - Gene copy number analysis for family data using semiparametric copula model. AB - Gene copy number changes are common characteristics of many genetic disorders. A new technology, array comparative genomic hybridization (a-CGH), is widely used today to screen for gains and losses in cancers and other genetic diseases with high resolution at the genome level or for specific chromosomal region. Statistical methods for analyzing such a-CGH data have been developed. However, most of the existing methods are for unrelated individual data and the results from them provide explanation for horizontal variations in copy number changes. It is potentially meaningful to develop a statistical method that will allow for the analysis of family data to investigate the vertical kinship effects as well. Here we consider a semiparametric model based on clustering method in which the marginal distributions are estimated nonparametrically, and the familial dependence structure is modeled by copula. The model is illustrated and evaluated using simulated data. Our results show that the proposed method is more robust than the commonly used multivariate normal model. Finally, we demonstrated the utility of our method using a real dataset. PMID- 19812788 TI - IMAGE: a new tool for the prediction of transcription factor binding sites. AB - IMAGE is an application tool, based on the vector quantization method, aiding the discovery of nucleotidic sequences corresponding to Transcription Factor binding sites. Starting from the knowledge of regulation regions of a number of co expressed genes, the software is able to predict the occurrence of specific motifs of different lengths (starting from 6 base pairs) with a defined number of punctual mutations. PMID- 19812789 TI - Identification and investigation of Drosophila postsynaptic density homologs. AB - AMPA receptors are responsible for fast excitatory transmission in the CNS and the trafficking of these receptors has been implicated in LTP and learning and memory. These receptors reside in the postsynaptic density, a network of proteins that links the receptors to downstream signaling components and to the neuronal cytoskeleton. To determine whether the fruit fly, Drosophila melanogaster, possesses a similar array of proteins as are found at the mammalian PSD, we identified Drosophila homologs of 95.8% of mammalian PSD proteins. We investigated, for the first time, the role of one of these PSD proteins, Pod1 in GluR cluster formation at the Drosophila neuromuscular junction and found that mutations in pod1 resulted in a specific loss of A-type receptors at the synapse. PMID- 19812790 TI - Identification of the RGG box motif in Shadoo: RNA-binding and signaling roles? AB - Using comparative genomics and in-silico analyses, we previously identified a new member of the prion-protein (PrP) family, the gene SPRN, encoding the protein Shadoo (Sho), and suggested its functions might overlap with those of PrP. Extended bioinformatics and conceptual biology studies to elucidate Sho's functions now reveal Sho has a conserved RGG-box motif, a well-known RNA-binding motif characterized in proteins such as FragileX Mental Retardation Protein. We report a systematic comparative analysis of RGG-box containing proteins which highlights the motif's functional versatility and supports the suggestion that Sho plays a dual role in cell signaling and RNA binding in brain. These findings provide a further link to PrP, which has well-characterized RNA-binding properties. PMID- 19812791 TI - Using a seed-network to query multiple large-scale gene expression datasets from the developing retina in order to identify and prioritize experimental targets. AB - Understanding the gene networks that orchestrate the differentiation of retinal progenitors into photoreceptors in the developing retina is important not only due to its therapeutic applications in treating retinal degeneration but also because the developing retina provides an excellent model for studying CNS development. Although several studies have profiled changes in gene expression during normal retinal development, these studies offer at best only a starting point for functional studies focused on a smaller subset of genes. The large number of genes profiled at comparatively few time points makes it extremely difficult to reliably infer gene networks from a gene expression dataset. We describe a novel approach to identify and prioritize from multiple gene expression datasets, a small subset of the genes that are likely to be good candidates for further experimental investigation. We report progress on addressing this problem using a novel approach to querying multiple large-scale expression datasets using a 'seed network' consisting of a small set of genes that are implicated by published studies in rod photoreceptor differentiation. We use the seed network to identify and sort a list of genes whose expression levels are highly correlated with those of multiple seed network genes in at least two of the five gene expression datasets. The fact that several of the genes in this list have been demonstrated, through experimental studies reported in the literature, to be important in rod photoreceptor function provides support for the utility of this approach in prioritizing experimental targets for further experimental investigation. Based on Gene Ontology and KEGG pathway annotations for the list of genes obtained in the context of other information available in the literature, we identified seven genes or groups of genes for possible inclusion in the gene network involved in differentiation of retinal progenitor cells into rod photoreceptors. Our approach to querying multiple gene expression datasets using a seed network constructed from known interactions between specific genes of interest provides a promising strategy for focusing hypothesis driven experiments using large-scale 'omics' data. PMID- 19812792 TI - [Evaluation of the validity of Diagnostic Interview Schedule for Children IV (DISC IV) in the city of Buenos Aires]. AB - The epidemiology of psychiatric disorders in children and adolescents has received little attention in Argentina. One of the problems related to the scarcity of such epidemiological research is linked to the lack of availability of diagnostic interview instruments which have been locally validated. OBJECTIVES: The object of the study was to conduct a validation study of the DISC IV (Spanish version), administered by lay interviewers in the City of Buenos Aires. METHODS: The sample was obtained from the Hospital de Ninos "Ricardo Gutierrez" in the City of Buenos Aires. Lay interviewers administered the DISC IV to 116 youngsters. Then psychiatrists re-administered the DISC IV a week later and immediately afterwards conducted a semi-structured diagnostic clinical interview. Participant in the sample ranged in age from 9 to 17. RESULTS: The sensitivity was 81.5% and the specificity 66.1%. The test-retest reliability was reasonable (Kappa 0.46 standard error 0.09). CONCLUSIONS: In general, the DISC administered by the non-professional interviewer was demonstrated to have the ability to discriminate between youngsters who suffer from psychiatric disorders and healthy youngsters. The confidence level was from moderate to good for the presence of a general psychiatric disorder as well as for disorders of specific states of mind, but for anxiety disorders and behaviour disorders the confidence level was poor. PMID- 19812793 TI - [Outpatient psychotherapy in suburban Buenos Aires: rate of use, costs, practices and patient characteristics]. AB - OBJECTIVE: The purpose of this study was to estimate the rate of use, costs and details of psychotherapeutic treatment in general population of suburban Buenos Aires along with patient demographic characteristics. METHODS: A survey was conducted between September and November 2007 among 899 inhabitants of 11 districts in greater Buenos Aires. RESULTS: During the previous month 14,2% of the population received psychotherapy treatment. Lifetime prevalence of psychotherapy was 42,8%. Use of psychotherapy was higher among women, the widowed, middle aged individuals and persons with higher socioeconomic status and levels of education. Almost 40% of the individuals in psychotherapy at the time of the survey reported receiving psychoanalytic treatment, 10% reported receiving cognitive-behavioral therapy while 41% was not aware of the type of psychotherapy received. Treatments were conducted mainly by psychologists (85%) and the majority consisted of individual psychotherapy in a private practice setting. Mean duration of a psychotherapy session was 52 minutes. A positive correlation was found between the duration of a session and its cost . There were no overall differences in the rate of use of psychotherapy between different areas of greater Buenos Aires, nor between these areas and previous estimates of prevalence in the city of Buenos Aires. CONCLUSIONS: Psychotherapy use in greater Buenos Aires is higher than in other countries. However, the rate of use is lower for certain socioeconomic and age groups. Further research is needed to determine whether the treatments observed are appropriate for the mental health needs of this population. PMID- 19812794 TI - [Alternatives to chronic hospitalization in mental health]. AB - It is not possible to work on alternatives to chronic institutionalization without taking into account a reform of the mental health care system. A great deal of experience has been accumulated and a lot of unsolved problems have arisen since the beginning of the deinstitutionalization in USA and Europe in the 60s. The aim was the closing of psychiatric hospitals, and so the accent was put on the physical place of treatment instead on the quality of the treatment or on the actual needs of the people involved. A review of the experiences is made and some clues are given to avoid the major problems and errors that have presented. Political decisions, experienced professionals, a stable and sufficient budget, modification of some laws, the creation of community institutions that fulfill the patient's needs before moving them, postgraduate training in rehabilitation, flexibility and creativity with empirical and scientific grounds are needed for a successful reform of the mental health care system. PMID- 19812795 TI - [Community assistance programme for children and adolescents with severe mental illness]. AB - The purpose of the following paper is to present an experience which links the production of knowledge resulting from academic research, with the transmission of said knowledge to the health system -with specific reference to mental health- by means of the implementation of the Community Assistance Programme for Children and/or Adolescents within the City of Buenos Aires. This programme is aimed at children and adolescents afflicted with severe psychological suffering, who consult the Mental Health Services in the public subsector of the City of Buenos Aires. It is the product of clinical institutional experience and of diverse investigations in health systems and services which showed a significant increase in the amount of consultations of children with severe and or complex mental health problems and which brought to light the lack of adequate sanitary response for these children by said sector. Based on the analysis and evaluation of the difficulties and obstacles in the functioning of the system a scheme was devised and implemented which aimed -from its beginning- to favour the possibilities of subjective constitution and integration into the community of these children and adolescents with severe mental deficiencies. By means of an interdisciplinary work consisting in the permanent training of human resources and the setup of intra and intersectional networks, it aims at facilitating accessibility and improving equality of opportunity, thus attempting to guarantee the right to a full protection of the health of children and adolescents. PMID- 19812796 TI - [Child sexual exploitation]. AB - Child Sexual Exploitation is a complex phenomenon in our country and the world; it dates back to an ancient past but it has a very recent conceptualization and specific approach. This article proposes a tour through this process as well as some inputs for its categorization, the attention to the affected subjects by the very design of public policies taken from a concrete institutional experience. PMID- 19812797 TI - [P.R.E.A. (Rehabilitation and Assisted Outpatient Care Program): an alternate experience to the "Asylum Logic"]. AB - The purpose of the following article is show the work that has been going on for the past nine years in the Assisted Discharge and Rehabilitation Program (PREA, from its Spanish initials), at the Esteves Hospital, Temperley, Province of Buenos Aires, Argentina. Our aim is to describe the historical and political context that gave rise to this program as well as the different activities carried out through its mechanisms to achieve the program's main objective: institutional discharge and resocialization of long-term patients with high chronicity in the psychiatric facility. This is the account of an experiment which, together with other similar ones already carried out in our country and elsewhere in the world, proves clearly that there are practical and sustainable alternatives to institutionalization and confinement for the treatment of serious mental disorders and chronicity; and that this is feasible within our province's current situation and our public health system. PMID- 19812798 TI - [Foster Home for children with mild and moderate behavioral problems]. AB - A methodology for working with children and adolescents with disruptive behavior is presented. This program has been in use since 1998 in a boarding home for boys in Buenos Aires. A common issue in this sample is the presence of non-functional families with high levels of violence between their members. PMID- 19812799 TI - [The origin of a modern view of the world: Tomaso Garzoni and his story about madness]. PMID- 19812800 TI - [The hospital of the incurable mad]. PMID- 19812802 TI - 2009 Canadian Cardiovascular Society/Canadian guidelines for the diagnosis and treatment of dyslipidemia and prevention of cardiovascular disease in the adult - 2009 recommendations. AB - The present article represents the 2009 update of the Canadian Cardiovascular Society guidelines for the diagnosis and treatment of dyslipidemia and prevention of cardiovascular disease in the adult. PMID- 19812803 TI - Angiographic results of the cobalt chromium Vision and Mini-Vision stents. AB - BACKGROUND: Drug-eluting stents (DES) have been shown to reduce repeat revascularizations compared with their bare-metal stent (BMS) platforms. Modern BMS may be associated with better angiographic results compared with the older BMS platforms. In the Basel Stent Kosten Effektivitats Trial (BASKET), target vessel revascularization after six months was nonsignificantly different between DES and BMS with clinical follow-up. OBJECTIVES: To evaluate angiographic results of the cobalt chromium Vision and Mini-Vision stents (Abbott Vascular, USA). METHODS: A total of 247 consecutive patients with 293 de novo lesions in native coronary arteries were treated with cobalt chromium Vision (n=184; stent diameter 2.75 mm to 4.0 mm) or Mini-Vision stents (n=109; stent diameter 2.0 mm to 2.5 mm), and scheduled for six months of angiographic follow-up. The primary end point was in-stent late loss after six months. RESULTS: Acute coronary syndromes were present in 83.4% (n=206) of patients. The preinterventional reference diameter of Vision stents was 2.70+/-0.34 mm and for Mini-Vision stents, it was 2.13+/-0.27 mm (P<0.001). Clinical and angiographic follow-up was 98.0% and 51.2%, respectively. In the Vision group, in-stent late loss was 0.64+/-0.67 mm and the binary restenosis rate was 17.9%. In the Mini-Vision group, in-stent late loss was 0.82+/-0.71 mm and the restenosis rate was 45.4%. No difference in occurrence of restenosis within the segments proximal or distal to the stent was observed. The restenotic pattern was predominantly focal with a short length of 7.9+/-4.4 mm. CONCLUSIONS: The use of the cobalt chromium Vision stent for the treatment of de novo lesions was associated with a low late loss and binary angiographic restenosis rate. PMID- 19812804 TI - Can we improve length of hospitalization in ST elevation myocardial infarction patients treated with primary percutaneous coronary intervention? AB - BACKGROUND: Patients with ST elevation myocardial infarction have traditionally been hospitalized for five to seven days to monitor for serious complications such as heart failure, arrhythmias, reinfarction and death. The Zwolle primary percutaneous coronary intervention (PCI) index is an externally validated risk score that has been used to identify low-risk primary PCI patients who can safely be discharged from the hospital within 48 h to 72 h. METHODS: The Zwolle score was retrospectively applied to all ST elevation myocardial infarction patients treated with primary PCI between April 2004 and February 2006 at a large Canadian teaching hospital. The goal was to characterize length of stay (LOS) in low-risk patients and to identify variables that correlate with patients who were hospitalized longer than expected. RESULTS: Data were collected on 255 patients. The mean LOS was 7.2+/-7.7 days (median 5.0 days [interquartile range 3.5 days]). A total of 179 patients (70%) had a Zwolle score of 3 or lower, identifying them as low risk. There was one death in the low-risk group (0.6% 30-day mortality) and 15 deaths in the higher-risk group (19.7% 30-day mortality), validating the Zwolle score in the population. A contraindication to early discharge was identified in 34 of the low-risk patients. Among the 144 remaining low-risk patients, the mean LOS was 5.1+/-3.3 days (median 4.0 days [interquartile range 3.0 days]). Only 8% were discharged within 48 h and only 28% within 72 h. It was determined that fewer patients were discharged on weekends and Wednesdays (when medical residents were away for teaching) than on other weekdays. LOS was longer among patients who were discharged on warfarin (7.6 days versus 4.6 days, P=0.006), and among patients who were transferred back to their presenting hospital rather than being discharged directly from the hospital where PCI was performed (5.6 days versus 4.0 days, P<0.001). CONCLUSIONS: Seventy-two per cent of low-risk primary PCI patients were hospitalized longer than 72 h. The following three factors were identified as correlating with prolonged LOS in this population: fewer discharges on days when there was less resident staffing; the use of warfarin at discharge; and transfer of patients back to their presenting hospital rather than discharging them directly from the PCI-performing hospital. A programmed approach to the identification and early discharge of low-risk patients could have significant cost savings and should be investigated prospectively. PMID- 19812805 TI - Prevalence and characteristics of home Internet access in patients with cardiovascular disease from diverse geographical locations. AB - BACKGROUND: Patients living outside of urban centres do not have access to the same level of care as patients in cities. The use of the Internet has been suggested as a possible resolution to this geographic inequity. OBJECTIVE: To identify the determinants of Internet use in patients with cardiovascular disease and the patterns of use. METHODS: Cardiac inpatients of an urban tertiary and a northern regional hospital in British Columbia were invited to participate. Patients who could not speak English or who had mental impairment were excluded. Consenting patients were interviewed regarding demographics and home Internet use. RESULTS: A total of 294 patients participated. The mean (+/- SD) age was 64.0+/-12.7 years. Most participants were men (68%) of European ancestry (77%) with some postsecondary education (57%). All geographical regions of British Columbia were represented. A total of 66% of patients had home Internet access. In rural areas, 47% of patients had access to the Internet (P=0.020 compared with nonrural areas). Eighty-four per cent of patients with Internet access had a high speed connection, 55% reported using the Internet daily and 23% used it more than once per week. Accessing health information from the Internet was reported by 70% of patients. CONCLUSION: These data indicate a high prevalence of Internet use among English-speaking cardiac patients and a strong desire to obtain health information using the Internet. Health care organizations can take advantage of the Internet to develop and evaluate the delivery of cardiac services to patients in their homes, particularly to patients in rural and remote communities. PMID- 19812806 TI - Exercise above the ischemic threshold and serum markers of myocardial injury. AB - BACKGROUND: Current guidelines for exercise training in coronary patients state that in the presence of exercise-induced ischemia, the heart rate during exercise should be at least 10 beats/min below the heart rate associated with an ST segment depression of 1 mm or greater. For patients with a relatively low ischemic threshold, this recommendation does not allow for a sufficient training stimulus. OBJECTIVE: To document the effects of a single session of exercise above the ischemic threshold on biochemical markers of myocardial injury in stable coronary patients with exercise-induced ischemia. Because creatine kinase (CK) and its MB isoenzyme (CK-MB) can both increase after exercise because of skeletal muscle injury, troponin T was also measured. METHODS: Twenty-one patients with documented coronary artery disease underwent two 20 min exercise sessions. The intensity of the first exercise training session was fixed at a heart rate below the ischemic threshold (ie, approximately 10 beats/min lower than the heart rate associated with the appearance of an ST segment depression of 1 mm or greater). The intensity of the second exercise session was fixed at a heart rate above the ischemic threshold. RESULTS: Blood test measurements at baseline, 6 h after and 24 h after the exercise sessions did not show any increase in total CK, CK-MB or troponin. The value of all measurements remained well below the lower limits associated with myocardial damage. CONCLUSION: A 20 min period of exercise above the ischemic threshold did not result in myocardial necrosis. PMID- 19812807 TI - Outcomes in heart failure patients referred for consideration of implantable cardioverter defibrillator for primary prophylaxis of sudden cardiac death: what are the risks of waiting? AB - BACKGROUND: Clinical trials have confirmed that implantable cardioverter defibrillators (ICDs) reduce mortality in patients with a left ventricular ejection fraction below 30%. The 'real-world' prognosis before ICD implantation in such patients is not known. The estimated risk of death is 0.8% per month, and this forms the basis for wait-time recommendations. OBJECTIVES: To determine the consequences of waiting for ICD implantation among heart failure patients eligible for primary prophylactic ICD. METHODS: The present retrospective study evaluated consecutive patients who were deemed eligible for primary prophylactic ICD implantation. Survival outcomes were tracked for patients who declined an ICD, those who accepted and received an ICD, and patients who accepted an ICD but died while waiting. RESULTS: Of 470 patients referred for evaluation, 218 were deemed eligible for an ICD. A total of 174 of 218 patients (79.8%) accepted an ICD; 39 (17.9%) declined, and five (2.3%) were deemed to be at too great a risk for the procedure. The mortality rate at two years among patients who accepted an ICD was 18.8% before ICD implantation and 12.2% after ICD implantation. Among patients who declined ICD implantation, the two year mortality rate was 5.3%. Among patients waiting for an ICD, five of 19 deaths were out-of- hospital sudden deaths. Of 12 patients who died after ICD implantation, there were no documented out-of-hospital sudden deaths. CONCLUSION: Consistent with current estimates, the mortality rate at two years among patients who accepted and were waiting for an ICD implant was 18.8%. After receiving an ICD, the mortality rate was 12.2% at two years. PMID- 19812808 TI - Factors related to time to admission to specialized multidisciplinary clinics in patients with congestive heart failure. AB - BACKGROUND: Congestive heart failure (CHF) is a common cause of hospitalization and has a poor prognosis. Specialized multidisciplinary clinics are effective in the management of CHF. OBJECTIVES: To measure time of admission to the specialized clinics and explore factors related to the time of admission to these clinics. METHODS: Patients who were newly admitted to one of six CHF multidisciplinary clinics in the province of Quebec were enrolled in the study. Data were collected from the common clinical database used at these clinics as well as from questionnaires administered to the patients. RESULTS: A total of 531 patients with a mean age of 65.9 years were enrolled. Only 26% were women. The median duration of disease before admission to the CHF clinic was 1.2 years. The majority of patients (62%) were referred by a cardiologist or an internist, while 24% were referred by other specialists, and 14% by general practitioners. One fifth of patients did not have regular follow-up for their CHF before being admitted to the clinic. Factors associated with shorter disease duration at admission to the clinic were referral by a specialist, not having regular medical follow-up for CHF, having a higher income and having visited the emergency room for CHF. CONCLUSION: There may be a need to improve dissemination of information regarding availability and benefits of CHF clinics and criteria for referral. PMID- 19812810 TI - A sweet tooth as the root cause of cardiac arrest. AB - A 71-year-old woman was admitted with hypotension and bradycardia. An electrocardiogram showed flattened T waves and increased U wave prominence, resulting in a long QT(U) syndrome. Her initial serum potassium level was 1.6 mmol/L (all other electrolytes, including magnesium, were normal). She suffered recurrent ventricular tachycardia and ventricular fibrillation arrest requiring direct current cardioversion and high-dose intravenous potassium chloride replacement. Systematic enquiry revealed that she had been constipated for a number of months and had resorted to consuming large quantities of liquorice on a daily basis for its laxative effects. Endocrinology review identified no primary abnormality of the renin- angiotensin- aldosterone axis, and the patient was diagnosed with hypokalemia secondary to liquorice overindulgence. Liquorice has a mineralocorticoid effect. If chronically consumed in large quantities, this effect may lead to severe depletion of whole-body potassium stores. The present case highlights a rare but important cause of hypokalemic cardiac arrest of which all acute care physicians should be aware. PMID- 19812809 TI - Effects of levosimendan versus dobutamine on left atrial function in decompensated heart failure. AB - BACKGROUND: Although the effects of levosimendan on the left ventricle (LV) have been studied, its effect on left atrial (LA) function is poorly understood, despite its key role in optimizing LV function. OBJECTIVE: To compare the effects of levosimendan and dobutamine on LA and LV function in patients with decompensated heart failure (DHF). METHODS: Seventy-four patients (mean [+/- SD] age 64+/-10 years) with DHF and an LV ejection fraction of 35% or lower were randomly assigned to receive levosimendan (n=37) or dobutamine (n=37). LA active emptying fraction, LA passive emptying fraction (PEF) and the ratio of mitral inflow early diastolic velocity to annulus velocity (E/e) were evaluated with pulsed wave and tissue Doppler imaging along with plasma B-type natriuretic peptide (BNP) level measurements before and after drug infusion. RESULTS: The ejection fraction was significantly increased in both groups. The levosimendan group had a greater decrease in BNP and a greater increase in active emptying fraction at 24 h compared with the dobutamine group. The PEF, E/e and deceleration time of the E wave were significantly improved in the levosimendan group, but not in the dobutamine group. Levosimendan- induced percentage change of BNP was significantly correlated with the percentage change of E/e and PEF (r=0.48 [P<0.005] and r=-0.38 [P<0.05], respectively). CONCLUSIONS: In patients with DHF, levosimendan and dobutamine both improve LV systolic function. However, levosimendan also improves LV diastolic function and LA performance in parallel with a greater improvement in neurohormonal activation compared with dobutamine. PMID- 19812811 TI - Acute myocardial infarction due to vasospasm induced by prostaglandin. AB - Prostaglandin E (PGE) is the preferred agent for second-trimester pregnancy termination. Hypotension, bradycardia, ventricular arrhythmias, myocardial infarction, cardiac arrest and death associated with PGE have been reported. A case of acute myocardial infarction due to coronary vasospasm induced by PGE is described in the present report. The diagnosis was confirmed by electrocardiography and coronary angiography. PMID- 19812812 TI - Whole-heart magnetic resonance imaging of isolated subpulmonary stenosis associated with hypertrophic cardiomyopathy. PMID- 19812815 TI - The hydrogen bond: a molecular beam microwave spectroscopist's view with a universal appeal. AB - In this manuscript, we propose a criterion for a weakly bound complex formed in a supersonic beam to be characterized as a 'hydrogen bonded complex'. For a 'hydrogen bonded complex', the zero point energy along any large amplitude vibrational coordinate that destroys the orientational preference for the hydrogen bond should be significantly below the barrier along that coordinate so that there is at least one bound level. These are vibrational modes that do not lead to the breakdown of the complex as a whole. If the zero point level is higher than the barrier, the 'hydrogen bond' would not be able to stabilize the orientation which favors it and it is no longer sensible to characterize a complex as hydrogen bonded. Four complexes, Ar2-H2O, Ar2-H2S, C2H4-H2O and C2H4 H2S, were chosen for investigations. Zero point energies and barriers for large amplitude motions were calculated at a reasonable level of calculation, MP2(full)/aug-cc-pVTZ, for all these complexes. Atoms in molecules (AIM) theoretical analyses of these complexes were carried out as well. All these complexes would be considered hydrogen bonded according to the AIM theoretical criteria suggested by Koch and Popelier for C-H...O hydrogen bonds (U. Koch and P. L. A. Popelier, J. Phys. Chem., 1995, 99, 9747), which has been widely and, at times, incorrectly used for all types of contacts involving H. It is shown that, according to the criterion proposed here, the Ar2-H2O/H2S complexes are not hydrogen bonded even at zero kelvin and C2H4-H2O/H2S complexes are. This analysis can naturally be extended to all temperatures. It can explain the recent experimental observations on crystal structures of H2S at various conditions and the crossed beam scattering studies on rare gases with H2O and H2S. PMID- 19812816 TI - Interactions and dynamics in electrolyte solutions by dielectric spectroscopy. AB - Despite its immense abilities to quantify many aspects of ion-ion and ion-solvent interactions, dielectric relaxation spectroscopy (DRS) has long been neglected as a tool for the investigation of the structure and dynamics of electrolyte solutions. The reasons for this are briefly discussed and it is shown that many of the difficulties associated with this technique have been overcome in recent years by technological developments. Representative applications of DRS to the investigation of ion solvation and ion association in electrolyte solutions of chemical, industrial, geochemical and biological interest, including room temperature ionic liquids and polyelectrolyte systems, are discussed. The advantages of linking DRS measurements to information obtained from other experimental techniques and from computer simulations are highlighted. PMID- 19812817 TI - Role of the substrate in electronic structure, molecular orientation, and morphology of organic thin films: diindenoperylene on rutile TiO2(110). AB - The results of our multitechnique investigation performed on diindenoperylene thin films deposited on rutile TiO2(110) show island growth, with crystallites nucleating preferentially along the [110] substrate crystallographic axis. The findings evidence that the films' properties at the interface are common to those found for a number of organic molecules deposited on the same substrate, revealing that the structural and morphological properties of organic thin films on rutile TiO2(110) are completely driven by its surface morphology. PMID- 19812818 TI - Ab initio modeling of protein/biomaterial interactions: competitive adsorption between glycine and water onto hydroxyapatite surfaces. AB - Both glycine and water exhibit a high affinity towards the hydroxyapatite HA surfaces. What happens when they are co-adsorbed at the HA (001) surface? B3LYP periodic calculations reveal that glycine displaces the pre-adsorbed water interacting directly with the HA surface. PMID- 19812819 TI - Response of the potential of a gold electrode to elastic strain. AB - We describe the measurement of the response, sigma, of the potential of a metal electrode to elastic strain under open circuit conditions. The experimental response exhibits a frequency dependence due to Faraday loss currents, which become negligible beyond 30 Hz. For a (111)-textured gold film the experimental value at higher frequency, sigma = -1.83 V, compares well with the intrinsic value of Au(111) predicted by recent ab initio computation. PMID- 19812820 TI - In search for an optimal methodology to calculate the valence electron affinities of temporary anions. AB - Recently, we have proposed an approach for finding the valence anion ground state, based on the stabilization exerted by a polar solvent; the methodology used standard DFT methods and relatively inexpensive basis sets and yielded correct electron affinity (EA) values by gradually decreasing the dielectric constant of the medium. In order to address the overall performance of the new methodology, to find the best conditions for stabilizing the valence state and to evaluate its scope and limitations, we gathered a pool of 60 molecules, 25 of them bearing the conventional valence state as the ground anion and 35 for which the lowest anion state found holds the extra electron in a diffuse orbital around the molecule (non valence state). The results obtained by testing this representative set suggest a very good performance for most species having an experimental EA less negative than -3.0 eV; the correlation at the B3LYP/6 311+G(2df,p) level being y = 1.01x + 0.06, with a correlation index of 0.985. As an alternative, the time dependent DFT (TD-DFT) approach was also tested with both B3LYP and PBE0 functionals. The methodology we proposed shows a comparable or better accuracy with respect to TD-DFT, although the TD-DFT approach with the PBE0 functional is suggested as a suitable estimate for species with the most negative EAs (ca.-2.5 to -3.5 eV), for which stabilization strategies can hardly reach the valence state. As an application, a pool of 8 compounds of key biological interest with EAs which remain unknown or unclear were predicted using the new methodology. PMID- 19812821 TI - Scaling form of viscosity at all length-scales in poly(ethylene glycol) solutions studied by fluorescence correlation spectroscopy and capillary electrophoresis. AB - We measured the viscosity of poly(ethylene glycol) (PEG 6000, 12,000, 20,000) in water using capillary electrophoresis and fluorescence correlation spectroscopy with nanoscopic probes of different diameters (from 1.7 to 114 nm). For a probe of diameter smaller than the radius of gyration of PEG (e.g. rhodamine B or lyzozyme) the measured nanoviscosity was orders of magnitude smaller than the macroviscosity. For sizes equal to (or larger than) the polymer radius of gyration, macroscopic value of viscosity was measured. A mathematical relation for macro and nanoviscosity was found as a function of PEG radius of gyration, R(g), correlation length in semi-dilute solution, xi, and probe size, R. For R < R(g), the nanoviscosity (normalized by water viscosity) is given by exp(b(R/xi)a), and for R > R(g), both nano and macroviscosity follow the same curve, exp(b(R/xi)a), where a and b are two constants close to unity. This mathematical relation was shown to equally well describe rhodamine (of size 1.7 nm) in PEG 20,000 and the macroviscosity of PEG 8,000,000, whose radius of gyration exceeds 200 nm. Additionally, for the smallest probes (rhodamine B and lysozyme) we have verified, using capillary electrophoresis and fluorescence correlation spectroscopy, that the Stokes-Einstein (SE) relation holds, providing that we use a size-dependent viscosity in the formula. The SE relation is correct even in PEG solutions of very high viscosity (three orders of magnitude larger than that of water). PMID- 19812822 TI - Ultrafast intermolecular energy transfer in heavy water. AB - We report on a study of the vibrational energy relaxation and resonant vibrational (Forster) energy transfer of the OD vibrations of D2O and mixtures of D2O and H2O using femtosecond mid-infrared spectroscopy. We observe the lifetime of the OD vibrations of bulk D2O to be 400 +/- 30 fs. The rate of the Forster energy transfer is measured via the dynamics of the anisotropy of the OD vibrational excitation. For a solution of 0.5% D2O in H2O, resonant energy transfer is negligible, and the anisotropy shows a single exponential decay with a time constant of 2.6 +/- 0.1 ps, representing the time scale of the molecular reorientation. With increasing concentration, the anisotropy decay becomes faster and non-exponential, showing the increased contribution of resonant energy transfer between the OD vibrations. We determine the Forster radius of the OD vibration of HDO in H2O to be r0 = 2.3 +/- 0.2 A. PMID- 19812823 TI - Experimental and calculated vibrational and electronic circular dichroism spectra of 2-Br-hexahelicene. AB - The vibrational circular dichroism (VCD) and IR absorption spectra of the (-) enantiomer of 2-Br-hexahelicene have been measured and interpreted by use of density functional theory (DFT) calculations. From time dependent DFT calculations we also interpret the electronic circular dichroism (ECD) spectra of 2-Br-hexahelicene. We compare the calculated IR, VCD and ECD spectra to the corresponding calculated data of hexahelicene and 2-aza-hexahelicene; for the last compound we also recorded the ECD spectra. Comparison with current literature allows an insight to be gained on the meaning and usefulness of some VCD features. PMID- 19812824 TI - The influence of reaction temperature on the oscillatory behaviour in the palladium-catalysed phenylacetylene oxidative carbonylation reaction. AB - This paper reports the influence of reaction temperature on the occurrence and characteristics of pH oscillations that are observed during the palladium catalysed phenylacetylene oxidative carbonylation reaction in a catalytic system (PdI2, KI, air, NaOAc) in methanol. Isothermal experiments were performed over the temperature range 10-50 degrees C. The experiments demonstrate that oscillations occur in the range 10-40 degrees C and that a decrease in reaction temperature results in an increase in the period and amplitude of the pH oscillations. Furthermore, it is observed that during oscillations at any specific temperature, the time taken for pH to increase from a minimum to a maximum value varies with respect to reaction time. However, the time required for the pH to fall from maximum to new minimum is approximately constant with respect to the reaction time and is a function of the reaction temperature. PMID- 19812826 TI - Decomposition of substituted alkoxy radicals--part I: a generalized structure activity relationship for reaction barrier heights. AB - An update and expansion of our readily applicable structure-activity relationship (SAR) for predicting the barrier height E(b) to decomposition by beta C-C scission of (substituted) alkoxy radicals is presented. Such alkoxy radicals are key intermediates in the atmospheric oxidation of volatile organic compounds, and a correct description of their chemistry is vital to the understanding of atmospheric chemistry; nevertheless, experimental data on these reactions remain scarce. The SAR is based on quantum chemical characterizations of a large set of alkoxy radicals, and accommodates alkoxy radicals with alkyl- (-R), oxo- (=O), hydroxy- (-OH), hydroperoxy (-OOH), alkoxy (-OR), alkylperoxy- (-OOR), nitroso- ( NO), nitro- (-NO2), nitrosooxy- (-ONO), and nitroxy- (-ONO2) functionalities, as well as 3- to 6-membered rings and some unsaturated side chains. The SAR expresses the barrier height to decomposition, E(b) = 17.9 kcal mol(-1) + Sigma N(s) x F(s), as a linear function of the number N(s) of these substituents on the relevant carbons, and the substituent-specific activities F(s) derived from the quantum chemical calculations, allowing facile predictions based solely on the molecular structure. For low barriers, < or = 7 kcal mol(-1), a simple curvature correction is required. The SAR-predicted barrier height E(b) can be used to predict the high-pressure rate coefficient for alkoxy decomposition k(diss) at or around 298 K. PMID- 19812825 TI - Electrochemical quartz crystal microbalance study of covalent tethering of carboxylated thiol to polyaniline for electrocatalyzed oxidation of ascorbic acid in neutral aqueous solution. AB - The electrochemical quartz crystal microbalance (EQCM) was used to study the electrosyntheses and electrochemical properties of two kinds of polyaniline (PANI)-thiol composite films in aqueous solutions, which were prepared by covalent binding of a thiol to the oxidized forms of PANI (PANI(post)-thiol, protocol A), and electropolymerization of aniline in the presence of a thiol (PANI(poly)-thiol, protocol B), respectively. The thiols involved were mercaptosuccinic acid (MSA), thioglycolic acid (TGA) and beta-mercaptoethanol (ME). The PANI(post)-thiol binding processes were monitored in situ with the EQCM, giving molar binding ratios (r, thiol vs. aniline unit of the polymer) of ca. 0.50 at saturation for these thiols. Both PANI(post)-thiol and PANI(poly) thiol composite films from the carboxylated thiols showed a controllable electroactivity of the PANI moiety in neutral even weakly alkaline phosphate buffer solutions (PBS), with maximum electroactivity roughly at r = 0.11 for PANI(post)-MSA or at r = 0.21 for PANI(post)-TGA. The PANI-thiol interaction was also supported by experiments of scanning electron microscopy, electrochemical surface plasmon resonance, Fourier transform infrared spectroscopy and ultraviolet-visible spectroscopy, and the interaction mechanism is briefly discussed. The PANI(post)-thiol and PANI(poly)-thiol composite films from the carboxylated thiols effectively electrocatalyzed the oxidation of ascorbic acid in pH = 7.3 PBS, and the PANI(post)-thiol exhibited electrocatalytic activity higher than the relevant PANI(poly)-thiol under our experimental conditions. The covalent anchoring of anionic thiol groups on the PANI backbone to prepare electroactive PANI in neutral solutions is conceptually new and may be extended to the development of new functional materials from many other conducting polymers and thiols for wide applications in catalysis, biosensing, molecular electronics, and so on. PMID- 19812827 TI - Coupled transport processes in responding membranes: the case of a single gradient. AB - We present a theoretical study of two coupled diffusion processes through a membrane which is able to respond to the presence of the diffusing substances with a phase change. The case we consider is when the direct driving force for one of the processes vanishes, i.e. when the chemical potential of one of the components, or some other intensive variable, is kept equal in the two reservoirs surrounding the membrane. The coupling of the two transport processes results in non-zero gradients inside the membrane also for the variable that is equal in the two reservoirs. There is a concomitant change in the phase coexistence compared to the behaviour expected if only considering the external conditions. The transport properties of the two phases within the membrane have to be different for this effect to occur, but otherwise the prerequisites are rather general. An analogous effect is expected for the case of coupled diffusion and heat conduction. Amphiphilic systems or biological membranes often show large changes in structure and transport properties due to small variations in thermodynamic variables such as temperature, the osmotic pressure of the solvent or the presence of cosolvents and cosolutes. The effects we find are therefore particularly relevant to these systems. PMID- 19812828 TI - Sorption strain as a packing phenomenon. AB - We employ Monte Carlo simulations in a semi-grand canonical ensemble to analyze the relation between sorption strains and the thermodynamic state of a confined fluid composed of "simple" fluid molecules that possess only translational degrees of freedom. Fluid molecules are confined to a slit-pore whose walls are composed of individual atoms distributed across the plane of each substrate according to the (100) structure of the face-centered cubic lattice. The substrates can be deformed to a certain extent on account of their own thermal energy and due to the interaction with the fluid molecules. We determine the phase diagram in both the bulk and in confinement for both rigid and deformable solid substrates. By using finite-size scaling concepts the location of the critical point is determined accurately. Our results indicate for the first time that the previously observed variation of sorption strains with the amount of adsorbed fluid material [G. Gunther et al., Phys. Rev. Lett., 2008, 101, 086104] is caused by packing effects (i.e. stratification of the confined fluid) but is largely independent of the precise nature of the thermodynamic state considered. PMID- 19812829 TI - Infrared spectroscopic studies of the heterogeneous reaction of ozone with dry maleic and fumaric acid aerosol particles. AB - Dicarboxylic acids, either directly emitted or formed in chemical processes, are found to be a significant component of tropospheric aerosols. To assess any potential chemical transformation of short unsaturated dicarboxylic acids in tropospheric heterogeneous chemistry, maleic and fumaric acid were selected as surrogates in this study. A novel aerosol flow tube apparatus is employed to perform kinetic studies of the oxidation of these organic compounds by gas-phase ozone. The system consists of a particle generation system, a vertically oriented glass flow tube and an infrared observation White cell with a Fourier transform infrared (FTIR) spectrometer for the detection system. A flow of single component organic aerosols with mean diameters ranging between 0.7 and 1.1 microm is introduced in a flow tube, in which the particles are subsequently exposed to a known concentration of ozone for a controlled period of time. A band assignment of infrared vibrational frequencies for dry maleic and fumaric acid aerosol spectra is presented. These studies are complemented with off-line analysis on the reaction products. The reaction exhibited pseudo-first-order kinetics on gas product formation, and the pseudo-first-order rate coefficients displayed a Langmuir-Hinshelwood dependence on gas-phase ozone concentration for both materials. By assuming a Langmuir-Hinshelwood behaviour, the following parameters were obtained: for the reaction of maleic acid aerosols, K(O3) = (3.3 + 0.5) x 10(-16) cm3 molecule(-1) and k(I)(max) = (0.038 + 0.004) s(-1); for the reaction of fumaric acid aerosols, K(O3) = (1.6 + 0.5) x 10(-16) cm3 molecule(-1) and k(I)(max) = (0.048 + 0.007) s(-1), where K(O3) is a parameter that describes the partitioning of ozone to the particle surface and k is the maximum pseudo-first order coefficient at high ozone concentrations. Apparent reactive uptake coefficients were estimated from the pseudo-first-order rate coefficient and a trend of decreasing uptake coefficients with increasing ozone concentrations was observed, in good agreement with literature values. PMID- 19812830 TI - Chiral recognition of 2-(3-benzoylphenyl)propionic acid (ketoprofen) by serum albumin: an investigation with microcalorimetry, circular dichroism and molecular modelling. AB - The interaction of enantiomeric ketoprofen (KP) with BSA and HSA was studied by isothermal titration calorimetry (ITC). Affinity constants and thermodynamic parameters for complexation in two main protein sites were determined. Affinity constants for both proteins are generally lower for S(+)- than for R(-)-KP. Large enthalpic contributions to Gibbs free energy are compensated by large negative entropic terms for S(+) in the BSA-subdomain IIIA and HSA-subdomain IIA. The lowest energy BSA complexes of both enantiomers were structurally characterized by combining molecular mechanics (MM) and molecular dynamics (MD) with circular dichroism (CD). Comparison of quantum mechanically calculated rotational strengths with the CD signals of the complexes supported the structures. These allowed to identify the main interactions of the KP enantiomers with surrounding amino acids at short distances, that limit significantly KP mobility in both sites. In the primary binding site S(+) is close to Tyr 409 in subdomain IIIA (Sudlow site II), and R(-) is close to Trp 212 and His 240 in subdomain IIA (Sudlow site I). The same sites are involved in the formation of 2:1 complexes. The equilibrium structures are characterized by marked geometrical distortion of KP. PMID- 19812831 TI - The role of the steps in the cleavage of the C-C bond during ethanol oxidation on platinum electrodes. AB - Ethanol oxidation has been studied on stepped platinum single crystal electrodes in acid media using electrochemical and Fourier transform infrared (FTIR) techniques. The electrodes used belong to two different series of stepped surfaces: those having (111) terraces with (100) monoatomic steps and those with (111) terraces with (110) monoatomic steps. The behaviors of the two series of stepped surfaces for the oxidation of ethanol are very different. On the one hand, the presence of (100) steps on the (111) terraces provides no significant enhancement of the activity of the surfaces. On the other hand, (110) steps have a double effect on the ethanol oxidation reaction. At potentials below 0.7 V, the step catalyzes the C-C bond cleavage and also the oxidation of the adsorbed CO species formed. At higher potentials, the step is not only able to break the C-C bond, but also to catalyze the oxidation of ethanol to acetic acid and acetaldehyde. The highest catalytic activity from voltammetry for ethanol oxidation was obtained with the Pt(554) electrode. PMID- 19812832 TI - Kinetic analysis of the rotation rate of light-driven unidirectional molecular motors. AB - The combination of a photochemical and a thermal equilibrium in overcrowded alkenes, which is the basis for unidirectional rotation of light-driven molecular rotary motors, is analysed in relation to the actual average rotation rates of such structures. Experimental parameters such as temperature, concentration and irradiation intensity could be related directly to the effective rates of rotation that are achieved in solution by means of photochemical and thermal reaction rate theory. It is found that molecular properties, including absorption characteristics and photochemical quantum yields, are of less importance to the overall rate of rotation than the experimental parameters. This analysis holds considerable implications in the design of experimental conditions for functional molecular systems that will rely on high rates of rotation, and shows that average rotation rates comparable to ATPase or flagella motors are within reach assuming common experimental parameters. PMID- 19812833 TI - Water-mediated conformer optimization in benzo-18-crown-6-ether/water system. AB - The conformation of benzo-18-crown-6-ether (B18C6) and its encapsulation of water molecules in a supersonic beam are investigated by laser induced fluorescence (LIF), UV-UV hole-burning, IR-UV double resonance (IR-UV DR), and resonance enhanced multiphoton ionization (REMPI) spectroscopy with the aid of density functional theory (DFT) calculations at the B3LYP/6-31+G* level. At least four B18C6 conformers and nine B18C6-(H2O)n (n = 1-4) clusters are identified in the supersonic beam. IR-UV DR spectra in the CH stretching region suggest that the four B18C6 conformers have different conformations from each other. In contrast, most of the nine B18C6-(H2O)n clusters have a very similar B18C6 conformation. IR UV DR spectra in the OH stretching region provide quite clear pictures of the hydration networks formed on B18C6. In all four B18C6-(H2O)1 isomers, the water molecule is H-bonded to the two O atoms adjacent to the benzene ring in "bidentate" and "bifurcated" manners. One of the four B18C6-(H2O)1 isomers exhibits a large population, and further hydration networks are preferentially grown on this specific isomer. PMID- 19812834 TI - The influence of support and particle size on the platinum catalysed oxygen reduction reaction. AB - A range of platinum deposits, equivalent thicknesses (delta) 0.2-2.5 nm, have been synthesised on carbon and reduced titania (TiO(x)) supports using physical vapour deposition on (10 x 10) arrays of electrodes. For delta < 1.0 nm, discrete platinum centres are formed and the TiO(x) supported platinum show two distinct characteristics: (a) a strong positive shift in the potential for the oxidation of monolayers of CO with decreasing loading of Pt leading to an inability to oxidise the CO on the lowest loadings and (b) a strong negative shift in the potential for the reduction of oxygen. Both observations can be understood in terms of an increase in the irreversibility of the Pt/PtO couple at such surfaces. The same trends, although significantly weaker, are seen with the carbon supported platinum, delta < 1.0 nm, and it is suggested that the Pt/PtO couple on carbon shows intermediate kinetics between Pt on TiO(x) and bulk Pt. These results have significant implications for understanding the mechanism of oxygen reduction on supported Pt catalysts and hence for the search for alternative supports to platinum for ORR electrocatalysts. PMID- 19812835 TI - Molecular mechanics force field-based map for peptide amide-I mode in solution and its application to alanine di- and tripeptides. AB - A molecular mechanics (MM) force field-based empirical electrostatic potential map (MM map) for amide-I vibrations is developed with the aim of seeking a quick and reasonable approach to computing local mode parameters and their distributions in solution phase. Using N-methylacetamide (NMA) as a model compound, the instantaneous amide-I normal-mode parameters (transition frequency and dipole) obtained at the level of MM force fields are converted to solution phase values by a four-site potential scheme, but without the need for quantum mechanical frequency computations of solute-solvent clusters as are required in constructing ab initio-based electrostatic potential or field maps. The linear IR line shape of the amide-I mode in NMA obtained from the frequency-time correlation function on the basis of the MM map are found to be comparable to those from the ab initio-based maps. Our results show that the amide-I local mode parameters are largely determined by the solvated peptide structure rather than by explicit solvent molecules, suggesting an inherent local structure sensitivity of the amide-I mode in solvated peptides. Applications to alanine di- and tripeptides are satisfactorily demonstrated, showing its usefulness as an alternative approach in providing vibrational parameters for the simulation of linear IR and 2D IR spectra of the amide-I modes in polypeptides. PMID- 19812836 TI - Structural and electronic properties of gold microclusters: assessment of the localized Hartree-Fock method. AB - We assess the localized Hartree-Fock (LHF) method for the simulation of the structural and electronic properties of gold microclusters. We compute the minimum-energy geometries, atomization energies, dipole moments, and single particle spectra for five gold clusters, Au(N), and their anions, Au(N)-, with N < or = 4. Calculations are performed with the LHF functional, with and without the addition of the Lee-Yang-Parr (LYP) correlation, and the results are compared with those obtained from other semilocal and hybrid functionals as well as from Hartree-Fock calculations. The LHF functional yields structural properties of similar quality to the other exchange-only methods, but superior molecular orbital energies are found. The LHF single-particle spectrum is free of the artifacts typical for HF and standard DFT calculations and in good agreement with available experimental reference data. The inclusion of correlation by the LYP functional produces a significant improvement of geometries and energetics, but has only a minor impact on the orbital energies. PMID- 19812837 TI - Para-hydrogen induced polarization in homogeneous phase--an example of how ionic liquids affect homogenization and thus activation of catalysts. AB - Para-hydrogen induced polarization (PHIP) is observed in an organic solvent-free homogeneous catalyst/substrate system using ionic liquids. The hydrogenation reactions are performed employing a Rh-catalyst/ionic liquid system without further organic solvents. The PHIP phenomenon is demonstrated for the ethyl acrylate system. Small amounts of ionic liquids (ILs) containing weakly coordinating anions such as [Tf2N]- act as homogenizing cosolvents for the catalyst in ethyl acrylate, leading to a dramatic activation of the catalytic process and a high PHIP signal enhancement. The achieved enhancement depends strongly on the viscosity of the ethyl acrylate/IL mixture. Furthermore, by deliberate choice of the IL constituents it was possible to design a model system exhibiting both sufficient catalyst solubility and biphasic liquid-liquid behavior. This shows that in ionic liquids PHIP enhancements followed by a fast quantitative removal of the para-hydrogenated product from the catalyst phase is in principle possible. The possible PHIP enhancement factors will depend strongly on the design of a suitable hydrogenation reactor. PMID- 19812838 TI - Resolutions of the Coulomb operator. Part III. Reduced-rank Schrodinger equations. AB - We consider a modified Schrodinger equation wherein the electron-electron repulsion terms r(ij)(-1) are approximated by truncated one-particle resolutions. Numerical results for the He atom and H2 molecule at the Hartree-Fock, second order Moller-Plesset, and configuration interaction levels show that the solutions of the resulting reduced-rank Schrodinger equations converge rapidly, and that even low-rank approximations can yield energies with chemical accuracy. PMID- 19812839 TI - Static and thermodynamic properties of low-density supercritical 4He-breakdown of the Feynman-Hibbs approximation. AB - We study the applicability of the semiclassical Feynman and Hibbs (FH) (second order or fourth-order) effective potentials to the description of the thermodynamic properties of quantum fluids at finite temperatures. First, we use path integral Monte Carlo (PIMC) simulations to estimate the thermodynamic/static properties of our model quantum fluid, i.e. low-density 4He at 10 K. With PIMC we obtain the experimental equation of state, the single-particle mean kinetic energy, the single-particle density matrix and the single-particle momentum distribution of this system at low densities. We show that our PIMC results are in full agreement with experimental data obtained with deep inelastic neutron scattering at high momentum transfers (D. Colognesi, C. Andreani, R. Senesi, Europhys. Lett., 2000, 50, 202). As expected, in this region of the 4He phase diagram, quantum effects modify the width of the single-particle momentum distribution but do not alter its Gaussian shape. Knowing the exact values of density, pressure and single-particle mean kinetic energy for our model quantum fluid, we investigate the limitations of the semiclassical FH effective potentials. We show that commonly used 'short-time' approximations to the high temperature density matrix due to Feynman and Hibbs can only be applied in a very limited range of the 4He phase diagram. We found that FH effective potentials reproduce the experimental densities of 4He at 10 K for Lambda/a < 0.45 (Lambda = 2.73 A denotes the thermal de Broglie wavelength, a = rho(-1/3) is the mean nearest-neighbor distance in the fluid and rho denotes fluid density). Moreover, semiclassical FH effective potentials are able to correctly predict the single particle mean kinetic energy of 4He at 10 K in a very limited range of fluid densities, i.e.Lambda/a < 0.17. We show that the ad hoc application of the semiclassical FH effective potentials for the calculation of the thermodynamic properties of dense liquid-like para-hydrogen (para-H2) adsorbed in nanoporous materials below 72 K is questionable. PMID- 19812840 TI - Water adsorption on the stoichiometric and reduced CeO2(111) surface: a first principles investigation. AB - We present a density functional theory investigation of the interaction between water and cerium oxide surfaces, considering both the stoichiometric and the reduced surfaces. We study the atomic structure and energetics of various configurations of water adsorption (for a water coverage of 0.25 ML) and account for the effect of temperature and pressure of the environment, containing both oxygen and water vapor, employing the ab initio atomistic thermodynamics approach. Through our investigation, we obtain the phase diagram of the water ceria system, which enables us to discuss the stability of various surface structures as a function of the ambient conditions. For the stoichiometric surface, we find that the most stable configuration for water is when it is bonded at the cerium site, involving two O-H bonds of hydrogen and oxygen atoms at the surface. If oxygen vacancies are introduced at the surface, which is predicted under more reducing conditions, the binding energy of water is stronger, indicating an effective attractive interaction between water molecules and oxygen vacancies. Water dissociation, and the associated activation energies, are studied, and the role of oxygen vacancies is found to be crucial to stabilize the dissociated fragments. We present a detailed analysis of the stability of the water-ceria system as a function of the ambient conditions, and focus on two important surface processes: water adsorption/desorption on the stoichiometric surface and oxygen vacancy formation in the presence of water vapor. A study of the vibrational contribution to the free energy allows us to estimate the effect of this term on the stability range of adsorbed water. PMID- 19812841 TI - A combined study based on experiment and molecular dynamics: perylene tetracarboxylate intercalated in a layered double hydroxide matrix. AB - This paper describes a combined experimental and theoretical simulation investigation on the photophysical properties, thermolysis, and orientation of 3,4,9,10-perylene tetracarboxylate (PTCB) intercalated Mg-Al-layered double hydroxide (PTCB/Mg-Al-LDH). UV-vis absorption and fluorescence spectroscopy show the existence of PTCB aggregates within the gallery of LDH, indicative of H-type (blue-shifted absorption band) and J-type dimers (red-shifted absorption band). In situ high-temperature X-ray diffraction (HT-XRD), thermogravimetry and differential thermal analysis (TG-DTA) and elemental analysis were used to study the thermal decomposition properties of PTCB/Mg-Al-LDH, and it was found that the decomposition temperature of the intercalated PTCB is lower than its pristine form (460 vs 565 degrees C), indicating that the strong pi-pi interaction among PTCB was weakened by the positively charged LDH host layers. Molecular dynamics (MD) calculations were employed to simulate the molecular arrangement and aggregation behavior of intercalated PTCB in the gallery of Mg-Al-LDH. The simulation results show that the intercalated PTCB anions exhibit a tendency from tilted to vertical orientation with respect to the layers as the interlayer water content increases, furthermore, the H and J-type dimer species are most likely populated under low and high hydration conditions, respectively. Moreover, the distribution of interlayer water molecules are also discussed. PMID- 19812842 TI - Towards nonlinear selection of reaction-diffusion patterns in presence of advection: a spatial dynamics approach. AB - We present a theoretical study of nonlinear pattern selection mechanisms in a model of bounded reaction-diffusion-advection system. The model describes the activator-inhibitor type dynamics of a membrane reactor characterized by a differential advection and a single diffusion; the latter excludes any finite wave number instability in the absence of advection. The focus is on three types of different behaviors, and the respective sensitivity to boundary and initial conditions: traveling waves, stationary periodic states, and excitable pulses. The theoretical methodology centers on the spatial dynamics approach, i.e. bifurcation theory of nonuniform solutions. These solutions coexist in overlapping parameter regimes, and multiple solutions of each type may be simultaneously stable. The results provide an efficient understanding of the pattern selection mechanisms that operate under realistic boundary conditions, such as Danckwerts type. The applicability of the results to broader reaction diffusion-advection contexts is also discussed. PMID- 19812843 TI - Tailoring the properties and the reactivity of the spinel cobalt oxide. AB - Pulsed spray evaporation chemical vapor deposition (PSE-CVD) was employed for the synthesis of cobalt-based spinel oxide thin films, Co(3-x)Fe(x)O4 with x = 0 1.56. XRD, Raman scattering and FTIR emission spectroscopy show that the normal spinel structure was retained for 0 < or = x < or = 0.65 by the selective insertion of Fe3+ in the octahedral sites. The spinel inversion was noticed above this range, whereas the insertion of Fe2+ was first indicated with x > or = 1. The room-temperature electrical resistivity of the thin films was controlled between 9 and 0.007 Omega cm by the adjustment of iron doping concentration. Furthermore an improvement of the thermal stability of the spinel was noticed upon doping by iron. The reducibility of the spinel in the presence of molecular hydrogen was efficiently adjusted by a shift of the reduction temperature by up to 110 degrees C upon the controlled insertion of iron in the octahedral sites of the spinel. The investigation of the catalytic oxidation of CO and ethanol over Co(3-x)Fe(x)O4 films with controlled structural modification enabled the confirmation of the Mars-van Krevelen mechanism for the oxidation of CO and the tight correlation between the selectivity of the conversion of ethanol to acetaldehyde and the abundance of surface basic sites. The controlled iron doping was demonstrated to be an efficient strategy to tune the reactivity and the selectivity of the cobalt-based spinel oxide. The doping-induced transition from normal to inverse spinel was observed to induce a clear discontinuity in the trend of all investigated physicochemical properties. PMID- 19812844 TI - A reversible switch for hydrogen adsorption and desorption: electric fields. AB - Implementation of hydrogen storage systems requires moderate bonding strength. However, this goal has remained a challenge, either due to the weak physisorption or extremely strong chemisorption. Here, we report on a new phenomenon, namely that H2 binding can be externally enhanced (or weakened) via superimposition of a positive (or negative) electric field. We demonstrate this concept using an 8-Li doped carbon nanotube. The calculated adsorption energy E(ad) = -0.58 eV/H2 under F = +0.010 au is 93.33% lower than that under 0.000 au (F indicates the field intensity). This is because the positive field produces an extra dipole moment. In contrast, E(ad) increases from -0.30 to -0.20 eV/H2 when F = -0.010 au. In view of the fact that storage systems are insensitive to small unexpected field fluctuations, the application of the electric field as a reversible switch makes practical sense. PMID- 19812845 TI - Solid-state NMR and computational studies of tetratolyl urea calix[4]arene inclusion compounds. AB - Solid-state guest dynamics of tetratolyl tetraurea calix[4]arene tetrapentylether dimeric capsules filled with different types of aromatic guests such as benzene d6, fluorobenzene-d5 and 1,4-difluorobenzene were studied. Upon inclusion, all guest moieties revealed complexation-induced shifts varying from 2.8 ppm to 5.1 ppm. All guest molecules were shown to undergo distinct motions, ranging from mere C6-rotations of benzene-d6 to (ill-defined) 180 degrees phenyl flips of fluorobenzene-d5. In all cases, dynamic heterogeneities were identified based on 2H lineshape deconvolution. In addition, by combination of both a computed nucleus independent chemical shift (NICS) map and explicit 19F and 2H ab initio DFT chemical shift calculations, the preferred orientation of the guest molecules within the host was derived. PMID- 19812846 TI - Quantum-chemistry calculations of hydrogen adsorption in MOF-5. AB - High concentrations of molecular hydrogen adsorption on MOF-5 were evaluated at the semiempirical PM6 (periodic and cluster) and ab initio MP2 (cluster) theoretical levels. From the semiempirical calculations, an uptake of 3.9% weight on the inorganic building unit of MOF-5 was estimated, in good agreement with a recent accurate estimation of 4.5-5.2%. Although PM6 allows a correct estimation of the maximum uptake, the adsorption energy was overestimated and hence ab initio calculations, including a correlation treatment at the MP2 level as well as corrections for basis set superposition error, were performed with full optimisation, including the 6-31G basis set, which rendered an adsorption energy (per hydrogen molecule) of -0.14 kcal mol(-1). The crucial role of the quality of the basis set, as well as the importance of simulating high hydrogen loading (resembling experimental measurements), are remarked. Single point calculations (using the 6-31G geometry) with improved basis sets 6-31G(d,p) and 6-31++G(d,p) yielded adsorption energies of -0.33 and -0.57 kcal mol(-1), the latter in reasonable agreement with a recent experimental estimation of -1.0 kcal mol(-1). The role of the intermolecular hydrogen interactions is highlighted in this study, since many previous computational studies were performed at low hydrogen loadings, far from the experimental uptake conditions. PMID- 19812847 TI - A mechanistic study of the low pressure pyrolysis of linear siloxanes. AB - Matrix isolation IR spectroscopy has been used to study the vacuum pyrolysis of 1,1,3,3-tetramethyldisiloxane (L1), 1,1,3,3,5,5-hexamethyltrisiloxane (L2) and 3H,5H-octamethyltetrasiloxane (L3) at ca. 1000 K in a flow reactor at low pressures. The hydrocarbons CH3, CH4, C2H2, C2H4, and C2H6 were observed as prominent pyrolysis products in all three systems, and amongst the weaker features are bands arising from the methylsilanes Me2SiH2 (for L1 and L2) and Me3SiH (for L3). The fundamental of SiO was also observed very weakly. By use of quantum chemical calculations combined with earlier kinetic models, mechanisms have been proposed involving the intermediacy of silanones Me2Si=O and MeSiH=O. Model calculations on the decomposition pathways of H3SiOSiH3 and H3SiOSiH2OSiH3 show that silanone elimination is favoured over silylene extrusion. PMID- 19812848 TI - Influence of Si distribution in framework of SAPO-34 and its particle size on propylene selectivity and production rate for conversion of ethylene to propylene. AB - To investigate the effect of SAPO-34 particle size (with a fixed Si mole fraction in its framework) and that of the Si mole fraction (in a SAPO-34 framework with fixed particle size) on propylene selectivity and production rate for the conversion of ethylene to propylene, SAPO-34 was prepared by hydrothermal synthesis using tetraethyl ammonium hydroxide or morpholine as a structural agent. The conversion of ethylene was carried out at 473 K using SAPO-34. The selectivity for propylene, the rate of propylene production, and the lifetime of the catalyst were strongly influenced by the catalyst crystal size. The SAPO-34 with a approximately 2.5 microm particle size had the highest selectivity for propylene (approximately 80%) up to a high conversion of ethylene (approximately 70%), while SAPO-34 with smaller particles had a longer catalyst lifetime, implying that catalyst deactivation was suppressed. The mole fraction of Si in the SAPO-34 framework with fixed particle size had little influence on the selectivity for propylene, indicating that the acid strength of SAPO-34 is independent of the Si mole fraction and all protons in SAPO-34 behave equivalently. Furthermore, the acid strength of protons determined by the measurements of NH(3)-TPD (temperature-programmed desorption) spectra did not depend on either the Si mole fraction or the SAPO-34 particle size. This result was also evident in the cracking rate of n-butane, which increased proportionally with increasing number of protons in SAPO-34.The number of protons generated by the incorporation of Si4+ into the SAPO-34 lattice increased proportionally, up to one Si atom introduced into every cage of SAPO-34, but did not continue to increase with further introduction of Si4+ into the lattice. PMID- 19812849 TI - Study of supercooled orientationally disordered binary solid solutions II: cyclohexyl derivatives, neopentanol and neopentylglycol. AB - The main aim of the present investigation is to see how various relaxation processes including the chair-chair transformation (as found by earlier researchers at room temperature in mechanical relaxation spectroscopy) in cyclohexane derivatives evolve as the temperature is lowered. For this purpose, four remarkable (two-component) solid solutions that are orientationally disordered are investigated, where the first three systems are hydrogen (H-) bonded pairs, and the fourth is a non-H bonded pair. The former group is the two component system of cyclohexanol (CHXOL) + 2,2-dimethyl-1-propanol or neopentanol (NPOL); cyclohexanol (CHXOL) + cycloheptanol (CHPOL) & neopentanol (NPOL) + neopentylglycol (NPGOL) systems, and the lone non H-bonded pair that has been studied is cyanocyclohexane (CNCH) and cyclohexylchloride (CHC). In all these cases, the liquid mixtures on cooling form orientationally disordered phases which are a solid solution of the corresponding pure phases. The feature is common to all the four systems studied here, but in CHXOL + CHPOL, the phase I of CHXOL beyond x(m) > or = 0.1 only forms a solid solution (designated as S(I')) with the phase I of CHPOL. In CNCH + CHC the solid phase is stable for the concentration range 0 < or = x(m) < or = 0.4 (without transition to any other phase). The above solid phase I (or I') has been investigated at low temperatures and for several concentrations, by means of dielectric spectroscopy and differential scanning calorimetry (DSC). Depending upon the concentration, this phase reveals a glass transition in the temperature range 116-150 K and associated with this is a pronounced relaxation process identifiable with the so called alpha-process. The dielectric spectra of this process is found to follow the Havriliak-Negami (HN) equation. In context of the binary system study here, the analysis of the various parameters obtained show an isomorphic relationship between the phases of the pure components through a continuous change of parameters. Another process of much smaller magnitude designated as the alpha' process was also found in systems consisting of cyclohexyl derivatives above the glass transition temperature T(g) which kinetically freezes around 170 K. This process interestingly, is also non-Arrhenius in nature, becomes increasingly weaker with increase in the second component, and may be identified with (axial) chair-(equatorial) chair transformation. In addition in all these systems, a weak high-frequency process and a clear sub-T(g) process, are found which are designated as the beta- & gamma-processes respectively. The beta-process may be identified with Johari-Goldstein (JG) or beta(JG) process as it is found to follow the predictions of the coupling model proposed by Ngai. However, the identification of the gamma-process with internal degrees of freedom are fraught with some problems in the interpretation of the experimental data that are highlighted. The kinetic freezing of the various dielectric processes have been critically examined in relation to the T(g) found in the DSC experiments. PMID- 19812852 TI - Inhibiting effects of Enterococcus faecium non-biofilm strain on Streptococcus mutans biofilm formation. AB - BACKGROUND AND PURPOSE: Enterococcus faecium is a normal bowel commensal and lactic acid bacterium that is rarely found in the oral cavity. This study investigated whether a non-pathogenic and non-biofilm strain of E. faecium functioned as a probiotic strain toward biofilm formation by Streptococcus mutans, which is an etiological agent for dental caries. METHODS: The effects of E. faecium on streptococcal biofilm was evaluated by absorbance of safranin stained biofilm at 492 nm in a 96-well microtiter plate. RESULTS: The E. faecium strain demonstrated cell-number-dependent inhibition of biofilm in dual cultures with 4 laboratory and 16 clinical S. mutans strains, as well as laboratory strains of Streptococcus sobrinus and Streptococcus sanguinis, in 96-well microtiter plates. The inhibiting effects of E. faecium were not dependent on the production of bacteriocin from streptococci and E. faecium, low pH after mix culture, or biofilm formation levels of S. mutans. A culture supernatant sample of more than 10 kDa from E. faecium showed direct inhibiting effects toward S. mutans biofilm formation. Treatment of heat, butanol, and phenol to a supernatant sample restored biofilm formation in culture of S. mutans with the sample. Moreover, the tendencies of inhibition levels by the supernatant sample were associated with those by bacterial cells of E. faecium to S. mutans strains. CONCLUSIONS: The E. faecium non-biofilm strain produced an inhibiting protein to streptococci biofilm formation, showed various susceptibilities to inhibit streptococcal biofilm, and acted as a probiotic bacterial inhibitor of streptococcal biofilm formation. PMID- 19812853 TI - Genetic fingerprinting and antimicrobial susceptibility profiles of Pseudomonas aeruginosa hospital isolates in Malaysia. AB - BACKGROUND AND PURPOSE: Pseudomonas aeruginosa is the third most common pathogen causing nosocomial infections. The objective of this study was to investigate the antimicrobial resistance profiles and genetic diversity of hospital isolates of P. aeruginosa and to investigate the presence of several resistance genes and integrons. METHODS: In this retrospective study, 48 clinical isolates of P. aeruginosa from 6 public hospitals in Malaysia were analyzed by antimicrobial susceptibility test and DNA fingerprinting techniques. RESULTS: Most of the P. aeruginosa isolates were resistant to tetracycline (73%) and chloramphenicol (60%) and, to a lesser extent, cefotaxime (40%), ceftriaxone (31%), cefoperazone (29%), ticarcillin (25%), piperacillin (23%), and imipenem (21%). Less than 20% of the isolates were resistant to ceftazidime, gentamicin, cefepime, ciprofloxacin, amikacin, piperacillin-tazobactam, and aztreonam (10%). Of the 48 isolates, 33 were multidrug resistant. Two isolates were extended-spectrum beta lactamase (ESBL) producers using the double-disk synergy test. However, polymerase chain reaction (PCR) failed to detect any common ESBL-encoding genes in all isolates, except for bla(OXA-10) in PA7 that was found to be part of a class 1 integron-encoded aacA4-bla(IMP-9)-catB8-bla(OXA-10) gene cassette. Using PCR, class 1 integron-encoded integrases were detected in 19% of the P. aeruginosa isolates. Repetitive extragenic palindrome-PCR generated 40 different profiles (F = 0.50-1.0) and enterobacterial repetitive intergenic consensus-PCR produced 46 profiles (F = 0.51-1.0). Pulsed-field gel electrophoresis with SpeI digested genomic DNA resulted in 45 different profiles (F = 0.50-1.00). CONCLUSIONS: Aztreonam appeared to be the most effective agent against multidrug resistant P. aeruginosa isolates. Sixty nine percent of the P. aeruginosa isolates analyzed were multidrug resistant and the isolates were genetically diverse. PMID- 19812854 TI - Gene cassette arrays, antibiotic susceptibilities, and clinical characteristics of Acinetobacter baumannii bacteremic strains harboring class 1 integrons. AB - BACKGROUND AND PURPOSE: Acinetobacter baumannii isolates containing class 1 integrons belong to different clones, but only a few strains are successful at causing infection. This study was conducted to compare the characteristics among these clones with different epidemicity. METHODS: Eighty eight bacteremic isolates of A. baumannii were collected in a medical center in Taiwan during a 3 year period. The gene cassettes and antibiotic susceptibilities of the bacterial isolates were delineated and the patients' characteristics were compared. RESULTS: Class 1 integrons were detected in 75 isolates (85.2%). Most of the isolates belonged to 2 major clones, but only 1 of the 2 clones caused outbreaks in several hospitals in Taiwan. Restriction analyses of variable regions of the integron revealed identical gene cassettes among isolates within the same clone. The cassette arrays of the 3 clones were aacA4, catB8, aadA1 (clone I, epidemic clone); dhfr XII, unknown open reading frame (orfF), aadA2 (clone II, endemic clone); and aacC1, 2 unknown open reading frames (orfX, orfX'), aadA1a (clone III). The epidemic and endemic strains were multidrug resistant, but the former presented a higher resistance rate to ampicillin-sulbactam. Infections with epidemic strains were significantly associated with prior use of cephalosporins, but didn't contribute to a higher mortality rate. CONCLUSIONS: Judicious use of cephalosporins and rapid identification using the integron typing method might be helpful for the prevention of further spread of strains with epidemic potential. PMID- 19812855 TI - Cryptococcal disease in patients with or without human immunodeficiency virus: clinical presentation and monitoring of serum cryptococcal antigen titers. AB - BACKGROUND AND PURPOSE: Cryptococcus neoformans is an encapsulated pathogenic yeast that causes a wide range of clinical manifestations. The serum cryptococcal latex agglutination test is a simple, rapid, and reliable diagnostic test for cryptococcosis. This study was performed to assess the clinical relevance of serum cryptococcal antigen (CRAG) titer in patients with cryptococcosis with or without human immunodeficiency virus (HIV). METHODS: From January 1999 to December 2007, 45 patients with a diagnosis of cryptococcosis made by culture and/or histopathology were enrolled in this retrospective study. Ten patients had HIV and 35 were not infected. RESULTS: Patients with HIV were more likely to have central nervous system (CNS) involvement than patients without HIV (100% vs 37.1%; p = 0.0005), higher serum CRAG titers (median, 1:1024 vs 1:64; p < 0.05), higher positive cerebrospinal fluid (CSF) CRAG (100% vs 37.1%; p = 0.0005), and higher CRAG titers in the CSF (median, 1:1024 vs 1:32; p < 0.001). Patients without HIV were more likely to have pulmonary involvement (62.9% vs 0%; p = 0.0005) and no underlying disease at diagnosis (42.9% vs 0%; p = 0.011). Serum CRAG titers among patients without HIV with CNS or pulmonary cryptococcosis declined during treatment and no relapse was noted when serum CRAG titers were 0.68). CONCLUSION: MSG (0.5% and 2%) does not guarantee a higher intake among elderly. The chemosensory heterogeneity of the elderly population requires more individual flavor enhancement to improve the dietary intake and sensory experience. PMID- 19812867 TI - Characterization of different groups of elderly according to social engagement activity patterns. AB - OBJECTIVE: The aim of this research was to segment older people in subgroups with similar social engagement activity patterns in order to better target public health interventions. DESIGN: Cross-sectional data, collected in 2005 by Dutch community health services (response 79%), from 22026 independently living elderly aged 65 or older were used. Cluster analysis was performed to derive subgroups with common social engagement activity patterns, which were compared for their self-perceived health, mental health, physical health, and loneliness. RESULTS: Among the independently living older people, five subgroups were identified with different patterns of social engagement activities: less social engaged elderly, less social engaged caregivers, social engaged caregivers, leisure engaged elderly, and productive engaged elderly. The subgroups differed significantly in social engagement activities, socio-demographics, and health (p < 0.001). The groups with the highest relative numbers of older people who were frequently engaged in leisure and productive-related activities, also included relatively more elderly with a good self-perceived health (85.8% versus 58.8%), mental health (91.3% versus 74.6%), physical health (97.7% versus 73.0%), and elderly who were not lonely (70.0% versus 52.0%) when compared to the least healthy subgroup. CONCLUSION: Older people could be segmented in subgroups based on similar social engagement patterns. Groups with elderly who were less socially engaged demonstrate to be possible target groups for public health interventions, given the relatively high shares of unhealthy older people among them. PMID- 19812868 TI - Validation of the Mini Nutritional Assessment short-form (MNA-SF): a practical tool for identification of nutritional status. AB - OBJECTIVE: To validate a revision of the Mini Nutritional Assessment short-form (MNA(R)-SF) against the full MNA, a standard tool for nutritional evaluation. METHODS: A literature search identified studies that used the MNA for nutritional screening in geriatric patients. The contacted authors submitted original datasets that were merged into a single database. Various combinations of the questions on the current MNA-SF were tested using this database through combination analysis and ROC based derivation of classification thresholds. RESULTS: Twenty-seven datasets (n=6257 participants) were initially processed from which twelve were used in the current analysis on a sample of 2032 study participants (mean age 82.3y) with complete information on all MNA items. The original MNA-SF was a combination of six questions from the full MNA. A revised MNA-SF included calf circumference (CC) substituted for BMI performed equally well. A revised three-category scoring classification for this revised MNA-SF, using BMI and/or CC, had good sensitivity compared to the full MNA. CONCLUSION: The newly revised MNA-SF is a valid nutritional screening tool applicable to geriatric health care professionals with the option of using CC when BMI cannot be calculated. This revised MNA-SF increases the applicability of this rapid screening tool in clinical practice through the inclusion of a "malnourished" category. PMID- 19812869 TI - Effect of a self-care program on oxidative stress and cognitive function in an older Mexican urban-dwelling population. AB - OBJECTIVE: To determine the effect of a self-care program on oxidative stress (OxS) and cognitive function in an older, Mexican, urban-dwelling population. DESIGN: A longitudinal and pre-experimental study was carried out in a sample of 79 older healthy, urban-dwelling individuals residing in Mexico City, (62 females and 17 males), of which 71 of them (59 women and 12 males) complied with the entire self-care program. MEASUREMENTS: We measured OxS, cognitive function, the Nagi Disability Scale of physical task functioning, and Instrumental Activities of Daily Living (IADL) prior to and after 2 years of intervention with an active aging program. RESULTS: All older persons adopted healthy, self-care-based lifestyles according to the active aging program to which they were trained, which was associated with a statistically significant improvement of OxS and cognitive function markers on comparing pre- and post-community intervention data. CONCLUSIONS: Our findings suggest that self-care-based healthy lifestyles programs can improve the oxidative stress and cognitive function in urban dwelling elderly population. PMID- 19812870 TI - Balneotherapy, prevention of cognitive decline and care the Alzheimer patient and his family: outcome of a multidisciplinary workgroup. AB - According to the latest forecasts of the INSEE - Institut National de la Statistique et des Etudes Economiques (National Statistics and Economic Studies Institute), ageing of the French population will increase between 2005 and 2050: whereas 20.8% of the population living in continental France reached the age of 60 years or more in 2005, this proportion would be of 30.6% in 2035 and 31.9% in 2050. In 2050, 22.3 million persons will have reached the age of 60 years or more compared to 12.6 million in 2005, increasing by 80% in a 45-year period. In line with the actual age pyramid, ageing is unavoidable, as those who will reach 60 years of age in 2050 are already born (in 1989 or before). This expansion will be most important between 2006 and 2035, when the numerous "baby-boom" generations born between 1946 and 1975, will reach these ages. In future years, lifespan improvement will only emphasize this increase. Even if life expectancy stabilizes at the 2005 level, the number of seniors reaching 60 years or more would still increase to 50% between 2005 and 2050. This issue is identical in all countries of the European Union. Ageing is a major risk factor for dementia that will considerably worsen in the next years, if no curative therapies are found. Today, 25 million persons in the world suffer from Alzheimer's disease (AD). In France, it is estimated that 860,000 persons are affected and that 225,000 news cases are annually diagnosed. After 75 years of age, more than 20% of women and 13% of men are concerned. Forecasts for the coming years are frightening. Considering ageing of the population, the number of Alzheimer's disease cases should raise to 1.3 million in 2020 (20 patients for 1000 inhabitants) ant 2.1 million in 2040 (30 patients for 1000 inhabitants). PMID- 19812871 TI - Domain-specific cognitive effects of tramiprosate in patients with mild to moderate Alzheimer's disease: ADAS-cog subscale results from the Alphase Study. AB - OBJECTIVES: Tramiprosate (homotaurine, ALZHEMEDTM) was recently investigated for its efficacy, safety and disease-modification effects in a Phase III clinical study in mild to moderate Alzheimer's disease (AD) patients (the Alphase study). The primary cognitive endpoint measure of that study was the Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS-cog). To characterize potential cognitive benefits of tramiprosate, the present study describes exploratory analyses performed on scores obtained from the specific ADAS-cog subscales in order to determine whether specific domains of cognition may be differentially affected by tramiprosate, which would not have been evident from the measure's total score. DESIGN: Multi-center, double-blind, randomized, placebo-controlled study. SETTING: 67 investigative sites in the United States and Canada. PARTICIPANTS: A total of 1,052 patients were randomized. INTERVENTIONS: Patients were randomized to receive twice a day Placebo (n=353), tramiprosate 100 mg (n=352) and tramiprosate 150 mg (n=347). MEASUREMENTS: ADAS-cog assessments were conducted every three months over the 78-week study period. Exploratory analyses were performed by comparing ADAS-cog subscale scores between Placebo and each active treatment arm at each visit. RESULTS: The findings of this analysis revealed statistically significant differences or statistical trends in favour of tramiprosate on six ADAS-cog subscales, namely Following Commands, Language Comprehension, Ideational Praxis, Object Naming, Remembering Test Instructions, and Spoken Language Ability. Differences in favor of Placebo were only observed on the Constructional Praxis subscale. CONCLUSION: This exploratory analysis suggests that tramiprosate may have some benefit on memory, language and praxis skills in mild to moderate AD individuals. Future clinical studies of tramiprosate should include specialized neuropsychological tests to validate its effects within these cognitive domains. PMID- 19812872 TI - The feasibility of serving liquid yoghurt supplemented with probiotic bacteria, Lactobacillus rhamnosus LB 21, and Lactococcus lactis L1A--a pilot study among old people with dementia in a residential care facility. AB - BACKGROUND: Constipation and body weight loss are prevalent among old people. Treatment with different species of lactic acid bacteria has been suggested for various forms of gastrointestinal disorders. OBJECTIVE, PARTICIPANTS AND INTERVENTION: This pilot study was performed to assess the feasibility of administering and consuming a drinkable yoghurt containing Lactobacillus rhamnosus LB 21 and Lactococcus L1A, and to evaluate the influence on bowel movements and body weight in 15 people with dementia, aged 65-95 years in residential care facility. DESIGN: A drinkable yoghurt (Verum Drickyoghurt) with added bacteria was served daily for 6 months in conjunction with ordinary food intake. Body weight, bowel movement frequency and consistency, food and fluid intake and time spent in various activities were recorded for two weeks pre intervention, and 3 times during the follow-up period. RESULT: This study showed that the yoghurt was easy to serve, with few side effects for the participants and that the various recording forms and diaries were easy for the staff to complete. The extra intake of yoghurt did not have any detectable beneficial effect on bowel movements. The overnight fast was almost 15 hours per day. Body weight decreased by 0.65 kg/month (95% confidence interval. 0.27-0.97). CONCLUSIONS: The present study design was feasible in this group of old people with dementia. The absence of any detectable effect of the supplement on constipation, but rather a considerable loss in body weight, indicate that further research is needed in a large randomised controlled trial, if associations between dementia, constipation and energy balance are to be understood. PMID- 19812874 TI - High BMI and waist circumference are associated with a high prevalence of comorbidities in older Americans Act programs in Georgia senior centers. AB - OBJECTIVE: This study characterized the relationship of body mass index (BMI) and high waist circumference (WC) with the prevalence of selected comorbidities among older adults receiving nutrition and wellness services from Georgia's Older Americans Act programs at senior centers. METHODS: Participants were a convenience sample (N = 759, mean age = 75 years, 81% female, 63% white, 36% black). Correction factors were applied to measured WC and to measured and self reported height and weight. RESULTS: The prevalence of several comorbidities showed striking relationships with WC and BMI, independent of age, gender, and race. CONCLUSION: The health burden of overweight/obesity is very high in these older adults creating an urgent need for evidence-based nutrition, physical activity programs, and therapeutic lifestyle counseling to prevent and manage weight-related comorbidities. PMID- 19812873 TI - Left ventricular mass, abdominal circumference and age: the Fels longitudinal study. AB - PURPOSE: To determine the relationship of abdominal circumference with increased left ventricular mass (LVM) from young adulthood into old age. METHODS: Cross sectional echocardiographic images were taken from 182 men and 220 women in the Fels Longitudinal Study 20 to 75 years of age to determine left ventricular mass. Left ventricular mass was divided by stature raised to the power of 2.7 (LVM/ht2.7) in order to minimize the impact of heart size variation from body size without overcompensating for the adverse effect of obesity. Abdominal circumference was measured and BMI calculated from stature and weight and categories of overweight, obesity and abdominal obesity were determined using published cut points. Regression models were used to describe the relationships of age, abdominal circumference, BMI and self-reported physical activity to LVM/ht2.7. RESULTS: Age, abdominal circumference and BMI were each positively and significantly related to an increased LVM/ht2.7 in men and women (p < 0.05). In the men, multivariate models indicated that abdominal circumference and BMI were both significantly related to an increased LVM/ht2.7, but the inclusion of BMI in these models for the women reduced the association of abdominal circumference and physical activity below significant levels. In the men, there was also a quadratic association of abdominal circumference with LVM/ht2.7 that was significant along with BMI and physical activity. Sex-specific logistic regressions with BMI and abdominal circumference obesity categories did not change or improve the initial findings in men or women. CONCLUSIONS: In women, increases in abdominal fatness as reflected in abdominal circumference at any age are linearly related to an increase in LVM/ht2.7, but the relationship of overall fatness as reflected in BMI with LVM/ht2.7 is stronger. In men, both abdominal fatness and overall fatness at any age are linearly related to an increase LVM/ht2.7. However, the significant curvilinear association of abdominal circumference and a linear association of BMI and physical activity with LVM/ht2.7 indicate the possible positive covariate relationship of overall muscle mass with LVM/ht2.7. This reflects the physiological changes with age and demonstrate, in part, the complexity of the interpretations of the inter associations of body composition, the cardiovascular system and the aging process, but the impact among the elderly and the known inadequacy of BMI at these ages remains an area for continued clinical study. PMID- 19812875 TI - The clinical characteristics of female patients with Fournier's gangrene. AB - INTRODUCTION AND HYPOTHESIS: Our aim was to review our experience with Fournier's gangrene in female patients. METHODS: A retrospective review of ten consecutive female patients with Fournier's gangrene was performed. Etiological and predisposing factors, causative microbiological organisms, and clinical outcome were investigated. RESULTS: Mean age of the patients was 52.7 years, and the mean duration of hospitalization was 17.6 days. The etiologic origin of the gangrene was anorectal, dermatological, and urogenital infection in 50%, 20%, and 10% of patients, respectively. All patients underwent aggressive surgical debridement and a diverting colostomy. Nine patients survived, and one patient died for an overall mortality rate of 10%. CONCLUSIONS: Fournier's gangrene occurred in females with a pattern similar to that in males. We believe that a colostomy is an integral part of management for patients requiring extensive debridement, especially if the infection arises in the anorectal region. PMID- 19812876 TI - Surgical management of vaginal mesh erosion: an alternative to excision. AB - HYPOTHESIS AND INTRODUCTION: Management of vaginal mesh erosion can be conservative, but in a large proportion of patients, excision/trimming of the mesh is needed. This paper describes a technique using an intervening vulval pad of fat graft between the vaginal epithelium and the mesh as an alternative to excision. MATERIALS AND METHODS: Three cases are presented where vaginal mesh erosion was treated using an intervening vulval pad of fat as a graft to cover polypropylene mesh erosion, and the technique is described. RESULTS: There were no intraoperative complications. All patients reported no further symptoms at follow-up, and examination revealed no further mesh erosions. CONCLUSION: The use of intervening graft of vulval fat can be an alternative to excision in the management of patients with vaginal mesh erosion. PMID- 19812878 TI - Toxic effect and biochemical study of chlorfluazuron, oxymatrine, and spinosad on honey bees (Apis mellifera). AB - Under laboratory conditions, the comparative effects of two insect growth regulators, chlorfluazuron and oxymatrine, and spinosad as a biopesticide were examined on honey bee workers (Apis mellifera L.). Separate groups of bees were left for 24 h to feed on 50% sucrose solution containing different concentrations of the tested insecticides, and the lethal concentration that caused 50% mortality (LC(50)) was estimated. The inhibitory effects on acetylcholinesterase (AChE) and adenosine triphosphatase (ATPase) activities as biochemical indicators were determined in vivo after 24 h in head, thorax, and abdomen of surviving bees obtained after treatments with a view to explore the possible mode of action of these compounds. Results indicated that exposure to spinosad showed toxicity to honey bees with LC(50) value of 7.34 mg L(-1), followed by oxymatrine (LC(50) = 10.68 mg L(-1)), while chlorfluazuron was the least acutely toxic of the tested compounds (LC(50) = 2,526 mg L(-1)). Oxymatrine and spinosad at the same tested concentrations (2.5, 5, 10, and 20 mg L(-1)) significantly inhibited AChE activity in different organs of honey bee workers, and high inhibition percentage was obtained with the enzyme isolated from the thorax. However, chlorfluazuron at 400, 1,000, 2,000, and 4,000 mg L(-1) caused high inhibition of AChE activity isolated from the head (39.65% and 44.22% at 2,000 and 4,000 mg L(-1), respectively). In addition, the toxic effects of the tested compounds on activity of ATPase indicated that spinosad caused the highest inhibitory effect in different organs compared with oxymatrine at the same concentrations, and high inhibition was found with ATPase isolated from the head. The results also indicated that oxymatrine was the least active compound for inhibition of AChE and ATPase. PMID- 19812877 TI - Colorectal and anal symptoms in women with urinary incontinence and pelvic organ prolapse. AB - INTRODUCTION AND HYPOTHESIS: Our aim was to determine the prevalence of colorectal and anal (CRA) symptoms in women with urinary incontinence and pelvic organ prolapse (UI/POP) in a predominantly Latina population. METHODS: We reviewed charts of women seen in the urogynecology clinic for UI/POP for those who completed the colorectal anal distress inventory-8 (CRADI-8) on their first visit. A detailed history was taken independent of the questionnaire. RESULTS: Two hundred sixty-five women completed the questionnaire; 94% were Latina; 89% completed the questionnaire in Spanish. Of the women, 88% indicated at least one CRA symptom: 60% reported needing to strain hard to have a bowel movement; 59% reported sensation of incomplete bowel emptying; 21% indicated incontinence of solid or liquid stool, and an additional 30% of women reported flatal incontinence only, for a total anal incontinence rate of 58%. CONCLUSION: CRA symptoms are highly prevalent among women with UI/POP who completed the CRADI-8 in a predominantly Latina patient population. PMID- 19812879 TI - Catheter ablation for right ventricular outflow tract ventricular tachycardia induced by fever in a 7-year-old girl. AB - We report a 7-year old girl with ventricular tachycardia exacerbated by fever. She experienced her first symptom of palpitations during an influenza infection, and electrocardiogram showed wide QRS tachycardia of a left bundle branch block inferior axis pattern. We performed an electrophysiological study (EPS) for diagnosis, but the wide QRS tachycardia could not be induced. The wide QRS tachycardia recurred during upper respiratory tract infection with fever and ceased with lidocaine administration. Wide QRS tachycardia is characteristic of ventricular tachycardia (VT) exacerbated by fever. VT was not able to be induced at a normal temperature on EPS; however, when her temperature exceeded 38 degrees C, VT was easily induced and was accompanied by decreased blood pressure. Radiofrequency catheter ablation was performed in the right ventricular outflow tract (RVOT) just under the pulmonic valve, with a fractionated potential 29 ms before onset of V1 QRS. VT was terminated approximately 4 s after RF application, and then VT could not be induced with or without high fever. This is the first report of successful ablation treatment in RVOT VT exacerbated by fever in a young child. PMID- 19812880 TI - VO2@RER1.0: a novel submaximal cardiopulmonary exercise index. AB - Maximal oxygen consumption (VO2max) is the "gold standard" by which to assess functional capacity; however, it is effort dependent. VO2@RER1.0 is defined when VO2 = VCO2. Between December 22, 1997 and November 9, 2004, 305 pediatric subjects underwent cycle ergometer cardiopulmonary exercise testing, exercised to exhaustion, and reached a peak respiratory exchange ratio > or = 1.10. Group 1 subjects achieved a peak VO2 > or = 80% of predicted VO2max; group 2 subjects achieved a peak VO2 < or = 60% of predicted VO2max; and group 3 subjects achieved a peak VO2 between 61 and 79% of predicted VO2max. Linear regression analysis was performed for VO2@RER1.0 as a function of predicted VO2 for group 1 subjects. A 2 SD regression line and equation was created. VO2@RER1.0 data from groups 2 and 3 were plotted onto the normative graph. Contingency table and relative-risk analysis showed that an abnormal VO2@RER1.0 predicted an abnormal peak VO2(positive-predictive value 83%, negative-predictive value 85%, sensitivity 84%, and specificity 84%). VO2@RER1.0 is a highly sensitive, specific, and predictive submaximal index of functional capacity. This submaximal index is easy to identify without subjectivity. This index may aid in the evaluation of subjects who cannot exercise to maximal parameters. PMID- 19812881 TI - Staged surgical approach in neonates with a functionally single ventricle and arch obstruction: pulmonary artery banding and aortic arch reconstruction before placement of a bidirectional cavopulmonary shunt in infants. AB - The success rate of right-heart bypass surgery in patients with a functionally single ventricle (f-SV) and systemic obstruction is low. In patients with a high risk of subaortic stenosis, we performed an initial step of pulmonary artery banding (PAB) and arch reconstruction before placing a bidirectional cavopulmonary shunt (BCPS) in infants with or without Damus-Kaye-Stansel (DKS) anastomosis. We assessed the success of right-heart bypass surgery. Between October 2003 and August 2008, we performed surgery in 19 neonates (median age 5 days) with f-SV and arch obstruction. Extended aortic arch anastomosis, with or without distal arch augmentation, was performed in 10 patients, and subclavian flap aortoplasty was performed in 9 patients. The circumference of the PAB was determined as the individual patient's body weight in kilograms plus 16.2 +/- 3.7 mm. Eighteen of 19 infants (95%) underwent successful BCPS placement at a median age of 7.8 months. DKS anastomosis was performed concomitantly during BCPS placement in 11 infants in whom subaortic stenosis was morphologically suspected but not demonstrated physiologically. As our first-stage operation, arch reconstruction plus PAB provided high success rates for right-heart bypass operations. This strategy is not leading, but it is a reliable approach for progression along a Fontan pathway. PMID- 19812882 TI - Successful two-stage correction of ventricular septal defect and patent ductus arteriosus in a patient with fixed pulmonary hypertension. AB - We report a case of a 6-year-old boy with fixed severe pulmonary artery hypertension secondary to a ventricular septal defect (VSD) together with a patent ductus arteriosus (PDA). As a preliminary step, PDA embolization was performed following therapy with inhaled prostacyclin over a period of 6 months. Further, the patient underwent successful surgical VSD closure. We postulate that a staged procedure with long-term prostaglandin therapy might be capable of reducing pulmonary artery resistance and permitting total correction in a patient once considered to have inoperable pulmonary arteriopathy. PMID- 19812883 TI - Congenital absence of the semimembranosus muscle: case report. AB - Congenital anomalies of the muscles of the lower extremity are rare. A case of complete absence of the semimembranosus muscle incidentally found with magnetic resonance imaging is reported. The patient was a 55-year-old female presenting with knee pain and no previous history of trauma. Clinical and imaging findings were consistent with meniscal tears. Two cases of uncommon distal insertion of the semimembranosus tendon are also described to illustrate the anatomical variations in this area. The anatomical and biomechanical relations between the semimembranosus muscle and the posteromedial corner of the knee are examined in this case report. PMID- 19812884 TI - Mandible size and morphology determined with CT on a premise of dental implant operation. AB - PURPOSE: This study aimed to clarify the size and morphology of the mandible, and to identify the location of the mandibular canal for the planning of the dental implantation. METHODS: CT data of 79 Japanese patients (52 male and 27 female) were analyzed. RESULTS: The means of the height and width ranged from 27.6 to 31.0 mm and from 10.5 to 15.8 mm, respectively. The height in male subjects was significantly greater than that in female subjects, and the width in male subjects was slightly but not significantly greater than that in female subjects. The morphology of the mandible was classified into three types. Type C (round) (59-61%) was the most common in the posterior region, followed by type A (lingual concavity) (36-39%), whereas type B (buccal concavity) (58-74%) and type C (17 36%) were the most common types in the anterior region. The distance from the mandibular canal to the alveolar crest, ranged from 15.3 to 17.4 mm. The anterior loop, was observed on 55% of all sides. CONCLUSIONS: The size and morphology of the mandible and the location of the mandibular canal were determined clearly using cross-sectional CT images. PMID- 19812885 TI - Micro-computed tomography study of the subchondral bone of the vertebral endplates in a porcine model: correlations with histomorphometric parameters. AB - PURPOSE: Subchondral bone (SCB) of the vertebral endplates (VEP) is the principal site of changes in vertebral trabecular microarchitecture secondary to intervertebral disc degeneration. However, the microstructure of this region has not yet been clearly characterized. METHODS: One thoracic and one lumbar vertebral unit (vertebra-disc-vertebra) was removed in nine pigs aged 4 months. Three samples (one central and two laterals) were taken from each VEP. Micro-CT examination and histomorphometric measurements of the subchondral trabecular bone of the VEP were carried out. Correlations between micro-CT and histological parameters were sought. RESULTS: Trabecular network was significantly denser [increased bone volume fraction (BV/TV) and trabecular number (Tb.N), decreased intertrabecular separation (Tb.Sp)] in the cranial endplates of the vertebral units. It was also significantly denser and less well organized [increased degree of anisotropy (DA)] in the centre of the VEP. The thickness of the cartilage endplate (CEP), SCB and growth cartilage were significantly lower in the centre of the VEP. There was a significant negative correlation between BV/TV, Tb.N and DA with the thicknesses of the CEP and SCB whereas Tb.Sp was positively correlated with these two parameters. CONCLUSION: We observed densification of the trabecular network in the centre of the VEP overlying the nucleus pulposus, partly related to thinner hyaline cartilage. Densification is associated with more anisotropic architecture that could cause lower mechanical strength in this area. This study provides new information on the microarchitecture of the SCB of the VEP which will make it possible to validate future models. PMID- 19812886 TI - Subgenual cingulate volumes in offspring of bipolar parents and in sporadic bipolar patients. AB - Decreased volumes of subgenual cingulate (SGC) have been reported primarily among familial bipolar patients, which is one of the hallmarks of an endophenotype. In order to investigate specificity of SGC volume abnormalities to familial mood disorders and to test whether SGC volumes represent an endophenotype for BD, we measured SGC volumes in young affected and unaffected relatives of bipolar patients (high-risk design) and in sporadic bipolar patients. We included 20 unaffected, 15 affected offspring of bipolar I or bipolar II parents, 18 controls, and 19 sporadic bipolar patients between 15 and 30 years of age. SGC volumes were measured on 1.5 T 3D anatomical MRI images using standard methods. We also combined the effect sizes from all published studies of sporadic patients with mood disorders (N = 61) and controls (N = 84) using random-effect models. We found comparable SGC volumes among unaffected, affected offspring of BD parents and controls (F = 0.7, df = 2; 50, P = 0.47). Likewise no SGC abnormalities were found between sporadic bipolar and control subjects (F = 2.31, df = 1; 34, P = 0.14). When combining all available data from sporadic patients, there were no differences in left (SDM 0.19, 95% CI -0.13 to 0.51) or right (SDM -0.11, 95% CI 0.47 to 0.26) SGC volumes between sporadic bipolar patients and controls. The limitations of the study are cross-sectional design and inclusion of both bipolar I and bipolar II probands. In conclusion, SGC volume abnormalities were absent in unaffected, affected relatives of bipolar patients as well as sporadic bipolar patients and thus did not meet criteria for endophenotype. PMID- 19812887 TI - Circulating tumor cells in gastrointestinal cancer. AB - Since the first report in the nineteenth century, there have been numerous reports on the isolation and characterization of circulating tumor cells (CTCs) in peripheral blood in patients with various carcinomas. In general, CTCs have been observed in the peripheral blood of cancer patients at very low concentrations of 10(-7)-10(-8) of normal peripheral blood cells. The characterization is of considerable biomedical interest in order to understand how these cells can travel via the blood stream to anatomically distant sites and form metastatic disease. Recent progress in molecular oncology enables us to detect the CTCs in blood with highly sensitivity and specificity, and several studies have indicated the prognostic value of CTC detection in patients with gastrointestinal cancers. Detection and measurement of CTCs in patients with gastrointestinal cancers such as colorectal, gastric, and pancreatic cancers can be useful as a promising tool for judging tumor stage, predicting the distant metastasis and patient survival, and monitoring the response to cancer therapy. Standard procedures for CTC detection have to be established, and the clinical relevance should be verified in large-scale clinical trials. However, CTC detection is suggested to provide useful information for the tumor staging and anticancer treatments in clinical practices in the near future. PMID- 19812888 TI - Development of a novel reflux-free bilioenteric anastomosis procedure by using a bioabsorbable polymer tube. AB - BACKGROUND: Bilioenteric anastomosis allows reflux of bowel fluid into bile ducts, resulting in reflux cholangitis. We sought to develop a reflux-free procedure of bilioenteric anastomosis endowed using an artificial bile duct (ABD). METHODS: The study was performed in four groups of pigs. In Group A, end to-side choledochoduodenostomy using the interposed ABD with its distal end wrapped by the duodenal seromuscular layer was performed. In Group B, conventional end-to-side choledochoduodenostomy using the ABD was performed. In Group C, conventional end-to-side choledochoduodenostomy was performed. In Group D, sham operation was performed. In each groups, 12 weeks later, animals were evaluated. RESULTS: Bile amylase levels were significantly lower in Group A (36.9 +/- 21.7 U/L) compared with Group B (469 +/- 140.9 U/L). No significant differences were observed in bile amylase levels between Groups A and D or between Groups B and C. On histology, the neo-bile duct in Group A was covered with the duodenal muscular layer at the orifice into the intestinal tract. CONCLUSION: This study suggests that the ABD is useful in the regeneration of extrahepatic bile ducts and that to bury the ABD in the duodenum can be a preventive measure against reflux of bowel fluid to bile ducts. PMID- 19812889 TI - Cutaneous vasculitis induced by TNF inhibitors: a report of three cases. AB - We describe 3 rheumatoid arthritis (RA) patients with anti-tumor necrosis factor (TNF) therapy-induced cutaneous vasculitis. Two cases were induced by infliximab and the other, in whom cutaneous vasculitis was found early at the start of therapy, was induced by etanercept. Skin biopsy was obtained in 2 patients, with histology-proven leukocytoclastic vasculitis. One patient spontaneously improved after cessation of the TNF inhibitor. Two patients required oral corticosteroid, the efficacy of which was observed to be excellent and rapid. PMID- 19812890 TI - Porcine noroviruses and sapoviruses on Korean swine farms. AB - Porcine noroviruses (NoVs) and sapoviruses (SaVs), which belong to the family Caliciviridae, have been considered potential zoonotic agents for human infection, and several cases have been reported in Asian countries. In this study, a total of 537 porcine fecal samples collected from 64 swine farms in Korea were tested. Among 537 samples, porcine NoVs were detected by semi-nested RT-PCR in ten samples (1.9%), and porcine SaVs were detected by RT-PCR in 60 samples (11.2%), showing their circulation in Korea. The porcine NoVs were genetically related to strains of genotypes 11 and 18, of genogroup II (GII) of the genus Norovirus. The porcine SaV strains were genetically related to the porcine enteric calicivirus Cowden strain and to the previously identified Korean porcine strains in genogroup III (GIII) of the genus Sapovirus. In no case was co infection with both NoV and SaV observed in one pig. This is the first report describing porcine NoVs identified in Korea. PMID- 19812891 TI - Familial hemifacial spasm and determinants of late onset. AB - The role of hypertension in the late onset of hemifacial spasm (HFS) is evaluated in a family, spanning four generations. Magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA) revealed a variable anatomical relationship between nervous and vascular structures in the symptomatic cerebello-pontine angle. In one case, showing neurovascular conflict (NVC), microvascular surgical decompression was followed by clinical resolution of HFS. Neuroimaging suggesting NVC was found in all symptomatic patients of the last two generations and in three younger subjects not affected by HFS. As a determinant for the late development of clinical expression is reviewed the role of arterial hypertension, detected few years before HFS appearing in all symptomatic subjects. The distribution of NVC in several members of the same family suggests a genetic susceptibility towards vascular anomaly. PMID- 19812892 TI - Leprosy: report of a case with severe peripheral neuropathy. AB - Leprosy (Hansen's disease) is a chronic granulomatous infectious disease, caused by Mycobacterium leprae, with cutaneous and neurological manifestations. Leprosy is very rare in Europe but some cases are reported, especially among people coming from endemic areas. Here, we report a case of Hansen's disease and emphasize the importance of a prompt diagnosis and treatment also in non-endemic areas. PMID- 19812893 TI - Chemopreventive doses of resveratrol do not produce cardiotoxicity in a rodent model of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC), one of the most lethal cancers, results in more than one million fatalities worldwide every year. In view of the limited therapeutic alternatives and poor prognosis of liver cancer, preventive control approaches, notably chemoprevention, have been considered to be the best strategy in lowering the present prevalence of the disease. Resveratrol, a naturally occurring antioxidant and antiinflammatory agent found in grapes and red wine, inhibits carcinogenesis with a pleiotropic mode of action. Recently, we have reported that dietary resveratrol significantly prevents chemically-induced liver tumorigenesis in rats. One of the mechanisms of resveratrol-mediated chemoprevention of hepatocarcinogenesis could be related to its antiinflammatory action through hepatic cyclooxygenase (COX-2) inhibition. Although several COX-2 inhibitors are known to exert chemopreventive efficacy, not all are considered ideal candidates for chemoprevention due to the risk of adverse cardiovascular events. Accordingly, the objective of the present study was to evaluate the role of resveratrol on cardiac performance during experimental hepatocarcinogenesis initiated with diethylnitrosamine and promoted by phenobarbital. Rats had free access to diet supplemented with resveratrol four weeks before the carcinogen injection and 14 weeks thereafter. The cardiotoxicity of resveratrol was assessed by monitoring the cardiac function using transthoracic echocardiography as well as Western blot analysis of cardiac tissue. Long-term dietary administration of resveratrol dose-dependently suppressed hepatic tumor multiplicity, the principal endpoint for evaluating the chemopreventive potential of a candidate agent. The chemopreventive effects of resveratrol were also reflected in histopathological assessment of hepatic tissues. Resveratrol did not exhibit any cardiotoxicity but rather improved the cardiac function in a dose-responsive fashion. Our results indicate that resveratrol-mediated chemoprevention of rat liver carcinogenesis is devoid of any adverse cardiovascular events. Resveratrol may be developed as a chemopreventive as well as therapeutic drug for human HCC. PMID- 19812894 TI - Fatty acid beta-oxidation in germinating Arabidopsis seeds is supported by peroxisomal hydroxypyruvate reductase when malate dehydrogenase is absent. AB - Peroxisomal malate dehydrogenase (PMDH) oxidises NADH produced by fatty acid beta oxidation during seed germination and seedling growth. Arabidopsis thaliana beta oxidation mutants exhibit seed dormancy or impaired seed germination and failure of seedlings to degrade triacylglycerol (TAG), but the pmdh1 pmdh2 null mutant germinates readily and degrades TAG slowly during seedling growth. We reasoned that in the pmdh1 pmdh2 mutant an alternative means of oxidising NADH operates to allow a slow rate of beta-oxidation, such as NADH and NAD(+) transport across the peroxisomal membrane or activity of another peroxisomal oxido-reductase. Here we show that peroxisomal hydroxypyruvate reductase (HPR) is present in germinating seeds and although knocking out HPR has little effect on germination and early seedling growth, when knocked out in combination with PMDH it exacerbates the pmdh1 pmdh2 phenotype. It greatly increases the proportion of dormant seeds and reduces the rate of seed germination. Seedlings have increased sucrose dependence and resistance to 2,4-dichlorophenoxybutyric acid (2,4-DB), and slower rate of TAG breakdown. When PMDH is absent, malate is lower in amount in germinating seeds and when HPR is also absent, serine (the immediate precursor of hydroxypyruvate) is much higher. These results indicate that HPR can oxidise NADH at sufficient rate in the absence of PMDH to support beta-oxidation and hence seed germination. We conclude that while HPR normally plays little role in seed germination our results support the growing body of evidence that peroxisomal NADH cannot be exported to the cytosol for oxidation but is oxidised by resident oxido-reductases. PMID- 19812896 TI - Candidacidal effects of Rev (11-20) derived from HIV-1 Rev protein. AB - Rev is an essential regulatory protein for HIV-1 replication. Rev (11-20) is known as the significant region regarding the function of a nuclear entry inhibitory signal (NIS) of Rev. In this study, anticandidal effects and mechanism of action of Rev (11-20) were investigated. The result exhibited that Rev (11-20) contained candidacidal activities. To understand target site(s) of Rev (11-20), the intracellular localization of the peptide was investigated. The result showed that Rev (11-20) rapidly accumulated in the fungal cell surface. The cell wall regeneration test also indicated that Rev (11-20) exerted its anticandidal activity to fungal plasma membrane rather than cell wall. The fluorescent study using 1,6-diphenyl-1,3,5-hexatriene (DPH) further confirmed the membrane disruption mechanism(s) of Rev (11-20). The present study suggests that Rev (11 20) possesses significant potential regarding therapeutic agents for treating fungal diseases caused by Candida species in humans. PMID- 19812895 TI - Non-redundancy within the RAS oncogene family: insights into mutational disparities in cancer. AB - The RAS family of oncoproteins has been studied extensively for almost three decades. While we know that activation of RAS represents a key feature of malignant transformation for many cancers, we are only now beginning to understand the complex underpinnings of RAS biology. Here, we will discuss emerging cancer genome sequencing data in the context of what is currently known about RAS function. Taken together, retrospective studies of primary human tissues and prospective studies of experimental models support the notion that the variable mutation frequencies exhibited by the RAS oncogenes reflect unique functions of the RAS oncoproteins. PMID- 19812897 TI - Metabolic engineering of Escherichia coli for the biological synthesis of 7-O xylosyl naringenin. AB - Flavonoids are a group of polyphenolic compounds that have been recognized as important due to their physiological and pharmacological roles and their health benefits. Glycosylation of flavonoids has a wide range of effects on flavonoid solubility, stability, and bioavailability. We previously generated the E. coli BL21 (DE3) Deltapgi host by deleting the glucose-phosphate isomerase (Pgi) gene in E. coli BL21 (DE3). This host was further engineered for whole-cell biotransformation by integration of galU from E. coli K12, and expression of calS8 (UDP-glucose dehydrogenase) and calS9 (UDP-glucuronic acid decarboxylase) from Micromonospora echinospora spp. calichensis and arGt-4 (7-O glycosyltransferase) from Arabidopsis thaliana to form E. coli (US89Gt-4), which is expected to produce glycosylated flavonoids. To test the designed system, the engineered host was fed with naringenin as a substrate, and naringenin 7-O xyloside, a glycosylated naringenin product, was detected. Product was verified by HPLCLC/MS and ESI-MS/MS analyses. The reconstructed host can be applied for the production of various classes of glycosylated flavonoids. PMID- 19812898 TI - A highly effective and long-lasting inhibition of miRNAs with PNA-based antisense oligonucleotides. AB - MiRNAs are non-coding RNAs that play a role in the regulation of major processes. The inhibition of miRNAs using antisense oligonucleotides (ASOs) is a unique and effective technique for the characterization and subsequent therapeutic targeting of miRNA function. Recent advances in ASO chemistry have been used to increase both the resistance to nucleases and the target affinity and specificity of these ASOs.Peptide nucleic acids (PNAs) are artificial oligonucleotides constructed on a peptide-like backbone. PNAs have a stronger affinity and greater specificity to DNA or RNA than natural nucleic acids and are resistant to nucleases, which is an essential characteristic for a miRNA inhibitor that will be exposed to serum and cellular nucleases.For increasing cell penetration, PNAs were conjugated with cell penetrating peptides (CPPs) at N-terminal. Among the tested CPPs, Tat modified peptide-conjugated PNAs have most effective function for miRNA inhibition. PNA-based ASO was more effective miRNA inhibitor than other DNA-based ASOs and did not show cytotoxicity at concentration up to 1,000 nM. The effects of PNA-based ASOs were shown to persist for 9 days. Also, PNA-based ASOs showed considerable stability at storage temperature. These results suggest that PNA based ASOs are more effective ASOs of miRNA than DNA-based ASOs and PNA-based ASO technology, compared with other technologies used to inhibit miRNA activity can be an effective tool for investigating miRNA functions. PMID- 19812899 TI - A tdcA mutation reduces the invasive ability of Salmonella enterica serovar typhimurium. AB - We previously observed that the transcription of some flagellar genes decreased in Salmonella Typhimurium tdcA mutant, which is a gene encoding the transcriptional activator of the tdc operon. Since flagella-mediated bacterial motility accelerates the invasion of Salmonella, we have examined the effect of tdcA mutation on the invasive ability as well as the flagellar biosynthesis in S. Typhimurium. A tdcA mutation caused defects in motility and formation of flagellin protein, FliC in S. Typhimurium. Invasion assays in the presence of a centrifugal force confirmed that the defect of flagellum synthesis decreases the ability of Salmonella to invade into cultured epithelial cells. In addition, we also found that the expression of Salmonella pathogenicity island 1 (SPI1) genes required for Salmonella invasion was down-regulated in the tdcA mutant because of the decreased expression of fliZ, a positive regulator of SPI1 transcriptional activator, hilA. Finally, the virulence of a S. Typhimurium tdcA mutant was attenuated compared to a wild type when administered orally. This study implies the role of tdcA in the invasion process of S. Typhimurium. PMID- 19812900 TI - RNAi suppression of RPN12a decreases the expression of type-A ARRs, negative regulators of cytokinin signaling pathway, in Arabidopsis. AB - The 26S proteasome is a 2-MDa complex with a central role in protein turn over. The 26S proteasome is comprised of one 20S core particle and two 19S regulatory particles (RPs). The RPN12a protein, a non-ATPase subunit of the 19S RP, was previously shown to be involved in cytokinin signaling in Arabidopsis. To further investigate cellular roles of RPN12a, RNAi transgenic plants of RPN12a were constructed. As expected, the 35S:RNAi-RPN12a plants showed cytokinin signaling defective phenotypes, including abnormal formation of leaves and inflorescences. Furthermore, RNAi knock-down transgenic plants exhibited additional unique phenotypes, including concave and heart-shape cotyledons, triple cotyledons, irregular and clustered guard cells, and defects in phyllotaxy, all of which are typical for defective cytokinin signaling. We next examined the mRNA level of cytokinin signaling components, including type-A ARRs, type-B ARRs, and CRFs. The expression of type-A ARRs, encoding negative regulators of cytokinin signaling, was markedly reduced in 35S:RNAi-RPN12a transgenic plants relative to that in wild type plants, while type-B ARRs and CRFs were unaffected. Our results also indicate that in vivo stability of the ARR5 protein, a negative regulator of cytokinin signaling, is mediated by the 26S proteasome complex. These results suggest that RPN12a participates in feedback inhibitory mechanism of cytokinin signaling through modulation of the abundance of ARR5 protein in Arabidopsis. PMID- 19812901 TI - Production of cellulosic ethanol in Saccharomyces cerevisiae heterologous expressing Clostridium thermocellum endoglucanase and Saccharomycopsis fibuligera beta-glucosidase genes. AB - Heterologous secretory expression of endoglucanase E (Clostridium thermocellum) and beta-glucosidase 1 (Saccharomycopsis fibuligera) was achieved in Saccharomyces cerevisiae fermentation cultures as an alpha-mating factor signal peptide fusion, based on the native enzyme coding sequence. Ethanol production depends on simultaneous saccharification of cellulose to glucose and fermentation of glucose to ethanol by a recombinant yeast strain as a microbial biocatalyst. Recombinant yeast strain expressing endoglucanase and beta-glucosidase was able to produce ethanol from beta-glucan, CMC and acid swollen cellulose. This indicates that the resultant yeast strain of this study acts efficiently as a whole cell biocatalyst. PMID- 19812902 TI - Depressed antioxidant status in pregnant women on iron supplements: pathologic and clinical correlates. AB - Iron (Fe) remains a commonly prescribed supplement in pregnancy. Its possible pathologic potential is either uncommonly considered or ignored. We determined the antioxidant status in pregnant women with and without Fe supplements. Fifty eight apparently healthy pregnant women on Fe supplements were selected for the study from the antenatal clinic of the University College Hospital, Ibadan, Nigeria. Fifty-five aged matched pregnant women who were not on Fe from various parishes of the Christ Apostolic Church, Ibadan (non-drug using Christian sect) were randomly selected as controls. Both groups were classified according to the trimesters of pregnancy. The gestational age in both pregnant women on Fe supplements and non-supplement pregnant women was similar. Fruit and vegetables consumption was higher in the supplement than in the non-supplement group (57.2% vs. 37.3%). Anthropometric indices, weight, height, and BMI, were also similar. But while the weight of the Fe supplement group decreased by nearly 3% in the third trimester, it increased by over 10% (p < 0.00) in the non-supplement group in the same period. Serum Fe level was significantly higher in the supplement than the non-supplement group (p < 0.001). In contrast, the levels of the antioxidants, ascorbic acid, copper (Cu), zinc (Zn), and bilirubin were all significantly decreased (p < 0.05, p < 0.001, p < 0.05, and p < 0.05, respectively). Uric acid level though also lower in the supplement group did not reach statistical significance (p > 0.05), while vitamin E was similar in both groups. There was relative stability of all antioxidants except uric acid, which declined from the first to the last trimester in the non-supplement group. The significantly higher Fe level in the second trimester was sustained in the third trimester though to a lesser degree (p < 0.05) and associated with significant decreases in the following antioxidant levels in the supplement group, ascorbic acid, bilirubin, Cu, and Zn (p < 0.02, p < 0.02, p < 0.02, and p < 0.001, respectively). Uric acid and vitamin E though lower in the supplement group were not significantly different. Remarkably, percentage changes between the first and third trimesters revealed that serum Fe increased by over 116% in the Fe supplement group, while it only increased by over 50% in the non-supplement group. This was associated with 23.50% decrease in ascorbate level (p < 0.003) in the supplement group, while it decreased by only 3.70% in the non-supplement group (p > 0.05). Again vitamin E decreased by 17.22% in the supplement group, while it decreased by only 7.30% in the non-supplement group during the period. Uric acid and bilirubin levels decreased by similar proportions during the period, while Zn decreased by 18.55% in the supplement group and by 14.86% in the non-supplement group. In contrast Cu increased by 7.20% in the supplement group, while it increased by only 2.96 in the non-supplement group. Additionally, all the antioxidants in the supplement group except vitamin E, viz, ascorbic acid, bilirubin, Cu, uric acid, and Zn, were significantly inversely correlated with serum Fe level (r - 0.299, p < 0.05, r - 0.278, p < 0.05, r - 0.383, p < 0.05, and r - 0.0369, p < 0.05). These data imply markedly depressed antioxidant status in the Fe supplement pregnant group with attendant oxidative stress (most probably pro-oxidant Fe-induced). This is associated with molecular and cellular damage as well as a number of pathologic and clinical correlates that underlie the exacerbation of morbidity and mortality in maternal and child populations, particularly in the developing countries. This appears to call for serious caution and prior evaluation of antioxidant and Fe status and during the use of Fe supplements in pregnancy for monitoring and prognostic purposes and to avert or ameliorate oxidative stress-induced pathologies in maternal and fetal systems. PMID- 19812903 TI - [Peripheral nerve blocks of the lower extremities. Clinical and practical aspects]. AB - Peripheral nerve blocks (PNBs) of the lower extremities are effective techniques for anesthesia and postoperative pain control. So far, these techniques have been used less frequently than PNBs of the upper limb. Nevertheless, growing awareness of complications of neuroaxial techniques, improved equipment and modern techniques for nerve localization have led to an increased use of PNBs of the lower limb. Anesthesiologists should be familiar with the anatomical basics and procedural details of these PNBs. They should also know the typical complications and side-effects and thoroughly inform patients about such potential problems. Continuous PNBs (perineural catheters) allow the benefits of PNBs to be prolonged into the postoperative period. Compared to continuous neuroaxial techniques continuous PNBs are equally effective for pain control but seem to be associated with fewer complications and side-effects. PMID- 19812904 TI - [Retroperitoneal bronchogenic cyst treated by laparoscopic surgery]. AB - The diagnosis of primary retroperitoneal cystic tumors is very infrequent in surgical pathology. We report the case of a 51-year-old woman presenting with an incidental left-sided retroperitoneal mass (32 x 24 mm in diameter) suspected of being an adrenal tumor. Intraoperatively the tumor was identified as a cystic lesion filled with mucous secretion and laparoscopically completely resected. The diagnosis was histopathologically confirmed as a bronchogenic cyst. In this article the laparoscopic removal of such a rare benign congenital aberration resulting from an abnormal budding of the tracheobronchial tree is presented. PMID- 19812905 TI - [Necrotizing fasciitis after "banal" back pain. An unusual course of a retrocoecal appendicitis and its sequellae]. AB - Necrotizing fasciitis is a life-threatening disease which can only be successfully treated by an interdisciplinary team. An immediate and radical debridement with opening of all compartments and debridement of the affected fascia is the basis for a successful therapy. We report about the treatment of a 21-year-old man who was taken to hospital due to "banal" back pain which was caused by a perforated appendicitis. In only 2 days necrotizing fasciitis developed which spread out over the complete right leg. PMID- 19812906 TI - [Complicated umbilical hernia in patients with decompensated liver cirrhosis. Concept for risk reduction of repair]. AB - Umbilical hernia repair is often accompanied by complications in patients with liver cirrhosis and ascites. In recent years we have been using the following concept for treating umbilical hernias in such patients: repair of the hernia by direct sutures and concomitant implantation of two large bore Robinson drainage tubes until the wound healing was completed within the next postoperative 10-14 days. During this time the reconstruction of the abdominal wall is in our opinion as robust that the ascites no longer represents a risk. Preconditions to perform this procedure were the best medicamentous treatment of ascites as ever possible and the perioperative administration of prophylactic antibiotics like gyrase inhibitors to avoid spontaneous bacterial peritonitis. Over a period of 10 years (01.01.1997-31.12.2006) we operated on 22 patients suffering from liver cirrhosis and ascites because of a complicated umbilical hernia (incarceration, irreponibility, skin ulceration, leackage of ascites). One group of patients (n=10) was treated by umbilical hernia repair with the concomitant implantation of two drainage tubes and the other group (n=12) by umbilical hernia repair without draining off the ascites. Morbidity and mortality were compared in both groups in a retrospective analysis. The postoperative morbidity could be reduced from 25% to 10% by using the drainage tubes as well as the rate of recurrent hernias in the drainage group. Due to these experiences we use the concept as standard in such patients and would like to recommend it further. However, we would like to initiate a prospective, randomized, at best multicenter trial for further validation. PMID- 19812907 TI - [Esophageal squamous cell carcinoma: pre-operative combined radiochemotherapy from a surgical oncological viewpoint]. AB - Esophageal squamous cell carcinomas (ESCCs) are an interdisciplinary challenge in terms of diagnosis, multimodal and/or surgical treatment procedures and also postoperative management as they are often associated with multiple comorbidities. The gold standard for a curative treatment approach is radical surgery, which is standardized and can be carried out with acceptable morbidity and mortality rates. ESCCs are usually diagnosed at locally advanced tumor stages and neoadjuvant treatment procedures are therefore used. Patients who respond to neoadjuvant therapy (responders) have a significantly better survival rate. The neoadjuvant studies that are currently available need to be critically assessed as they do not include response as an end point. In this context the WHO clinical response evaluation that has been used up to now is questionable. The histopathological findings with percentage proportions of residual tumor cells represent the gold standard for evaluating the response. Positron-emission tomography as a response criterion is predictive for the histopathological response and survival. ESCC patients who are classified as non-responders do not appear to benefit from surgical resection. In the future the results of a response evaluation and of pretreatment molecular biological tests could have a place in the process of pretherapeutic and peritherapeutic oncological decision making in patients with ESCCs. PMID- 19812909 TI - A statistical approach for optimization of polyhydroxybutyrate production by Bacillus sphaericus NCIM 5149 under submerged fermentation using central composite design. AB - The aim of this work was to statistically optimize the cultural and nutritional parameters for the production of polyhydroxybutyrate (PHB) under submerged fermentation using jackfruit seed hydrolysate as the sole carbon source. On the basis of results obtained from "one variable at a time" experiment, inoculum age, jackfruit seed hydrolysate concentration, and pH were selected for response surface methodology studies. A central composite design (CCD) was employed to get the optimum level of these three factors to maximize the PHB production. The CCD results predicted that jackfruit seed hydrolysates containing 2.5% reducing sugar, inoculum age of 18 h, and initial medium pH 6 could enhance the production of PHB to reach 49% of the biomass (biomass 4.5 g/l and PHB concentration 2.2 g/l). Analysis of variance exhibited a high coefficient of determination (R(2)) value of 0.910 and 0.928 for biomass and PHB concentration, respectively, and ensured that the quadratic model with the experimental data was a satisfactory one. This is the first report on PHB production by Bacillus sphaericus using statistical experimental design and RSM in submerged fermentation with jackfruit seed hydrolysate as the sole source of carbon. PMID- 19812908 TI - [Principles of neoadjuvant therapy]. AB - Neoadjuvant therapy is now an integral part in the treatment of locally advanced cancer of the esophagus and stomach. The mechanisms of action comprise downsizing of the primary tumor, improvement of the T and N categories and the earliest possible eradication of micrometastases. Several controlled clinical studies could show that these principles can indeed augment the rate of curative (R0) resections and can reduce the likelihood of systemic recurrences. As a consequence overall survival is improved by neoadjuvant therapy. This article summarizes how the basic principles of neoadjuvant therapy translate into clinical practice and gives a review of current developmental perspectives in this field. PMID- 19812910 TI - Effect of fill time on the performance of pilot-scale ASBR and AnSBBR applied to sanitary wastewater treatment. AB - Many lab-scale studies have been carried out regarding the effect of feed strategy on the performance of anaerobic sequencing batch reactors (ASBR); however, more detailed pilot-scale studies should be performed to assess the real applicability of this type of operation. Therefore, the objective of this work was to assess the effect of feed strategy or fill time in a 1-m(3) mechanically stirred pilot-scale sequencing batch reactor, treating 0.65 m(3) sanitary wastewater in 8-h cycles at ambient temperature. Two reactor configurations were used: one containing granular biomass (denominated ASBR) and the other immobilized biomass on polyurethane foam as inert support (denominated anaerobic sequencing batch biofilm reactor (AnSBBR)). The reactors were operated under five distinct feed strategies, namely: typical batch and fed-batch for 25%, 50%, 75%, and 100% of the cycle length. Stirring frequency in the ASBR was 40 rpm with two flat-blade turbine impellers and 80 rpm in the AnSBBR with two helix impellers. The results showed that both the ASBR and AnSBBR when operated under typical batch, fed-batch for 50% and 75% of the cycle length, presented improved organic matter removal efficiencies, without significant differences in performance, thus showing important operational flexibility. In addition, the reactors presented operation stability under all conditions. PMID- 19812913 TI - Bulk flow and diffusion revisited, and clinical applications. AB - The first Klatzo-Lecture pays homage to an exceptional academician, scientist and teacher. The author spent nearly 1 year in Klatzo's laboratory at the NHI in Bethesda, and the first part of results presented here originate directly from this collaboration. It was shown that following cortical injury, movement of edema fluid into the tissue occurs by bulk flow, and that the driving force is a small tissue pressure gradient. Resolution of edema fluid is achieved by clearance into the ventricular and subarachnoid CSF, is enhanced in the presence of pressure gradients and is supported by re-absorption into capillaries. Using appropriate techniques, the formation rate as well as clearance of edema into CSF and tissue resorption could be determined in human brain metastases and malignant gliomas. Three examples of clinical applications based on the discussed mechanisms are presented: a. Fluorescence-guided surgery of gliomas is based on the accumulation of 5-ALA in tumour cells; there being enzymatically converted to PP-IX, a compound with deep red fluorescence. This fluorescence is used for the more accurate surgical removal of gliomas. b. Radioimmunotherapy of gliomas uses an anti-tenascin antibody, coupled with a nuclide, administered postoperatively into the tumour cavity, from where it diffuses into tissue, couples to the receptor at the glioma cells. Then the isotope destroys the tumour cells. c. Convection-enhanced delivery is based on the interstitial infusion of an appropriate cytotoxic drug into the white matter at low pressure. Thus, the method employs bulk flow, distributes a drug in a larger tissue volume and eventually achieves drug concentrations greater than systemic levels. Experimental studies and clinical results are presented for all three clinical applications.I am very grateful to Z. Czernicki and the organizing group for being offered the great honour of presenting the first Igor Klatzo Lecture. In this report first previous results of bulk flow and diffusion in the development and resolution of brain edema will be revisited, then some recent examples will be shown as to how this knowledge of diffusion and bulk flow can be transferred into clinical applications.A great part of the work on bulk flow and diffusion was done during a stay in I. Klatzo's laboratory in Bethesda in 1973/1974 (Fig. 1). Since then a long collaboration developed with I. Klatzo and M. Spatz. Due to given limits, I will concentrate on the studies of our group. Unfortunately it will not be possible to mention all the important groups who have contributed by essential studies. PMID- 19812914 TI - Experimental cerebral ischemia: the contribution of the Bethesda Group. AB - Igor Klatzo started his research on cerebral ischemia at the NIH in the 1960s. The mechanism that produces the blood-brain barrier change after ischemia was a focus of interest in Klatzo's experiments, which used larger mammals. Studies using Mongolian gerbils, started by U. Ito, resulted in several important findings, including observation of the maturation phenomenon in 1975. Using newly developed ischemia models, the mechanism of postischemic neuronal/tissue injury was extensively studied. The cumulative effect was observed after repetitive cerebral ischemia. The protective mechanism of cortical spreading depression after global ischemia was investigated. Projects including in vitro studies of human brain endothelial cells and mucosal tolerance to E-selectin were performed in the Stroke branch after Klatzo retired from the NIH. Klatzo published a biography of Cecil and Oskar Vogt after retirement. He passed away in May 2007 in Gaithersburg, Maryland a few months after he completed the first part of his autobiography. PMID- 19812915 TI - Past and recent BBB studies with particular emphasis on changes in ischemic brain edema: dedicated to the memory of Dr. Igor Klatzo. AB - The blood-brain barrier (BBB) functions to protect the environment of the brain through endothelial cells and their interactions with other cells and components of the cerebral vasculature and the brain parenchyma. Alterations in the BBB as a result of injuries (i.e., brain ischemia and traumatic brain injury) play a crucial role in the pathophysiological response.The following is a brief review of the BBB and the mechanisms by which its cellular elements participate in barrier disruptions such as those associated with ischemia and resulting brain edema formation. PMID- 19812916 TI - Increase in activity of neutrophils and proinflammatory mediators in rats following acute and prolonged focal cerebral ischemia and reperfusion. AB - PURPOSE: It has been proposed that the immune system is activated during ischemic cerebral events and that brain damage caused by ischemia is increased by this immune activity. Neutrophils (PMNs) are one of the first factors in the chain of reactions of the immune system during focal cerebral ischemia. Experimental and clinical studies have emphasized the important role of proinflammatory cytokines such as interleukin-1beta (IL-1beta) and tumor necrosis factor (TNFalpha), in addition to vasoactive peptide and endothelin-1 (ET-1), in the formation of cerebral ischemia. MATERIAL AND METHODS: The experiments were carried out using Wistar rats that were divided into four groups: three experimental groups (acute and prolonged focal cerebral ischemia and following reperfusion) and one control group (sham). Focal cerebral ischemia was induced by the intraluminal surgical suture method. The oxidative activity of PMNs was measured after stimulation with phorbol myristate acetate, a protein kinase C activator (luminol enhanced chemiluminescence). The concentration of IL-1beta and TNFalpha in rat lymphocyte culture after stimulation with CSF was determined using commercial ELISA kits. The plasma concentration of ET-1 was determined using commercial kits with the RIA method. RESULTS: We confirmed a statistically significant increase in the oxidative activity of PMNs in rats with acute focal cerebral ischemia (p < 0.00001), prolonged ischemia (p < 0.001) and reperfusion (p < 0.05). An increase in IL-1beta and TNFalpha in lymphocytes following CSF stimulation was observed in the group with prolonged ischemia and in the group with reperfusion after transient ischemia (p < 0.05 for both). An increase in plasma ET-1 concentration was observed with acute and prolonged focal cerebral ischemia (p < 0.05 and p < 0.01, respectively). CONCLUSIONS: Our results show that acute and prolonged focal cerebral ischemia and reperfusion induce statistically significant increases in the oxidative activity of PMNs. The concentration of proinflammatory mediators (IL-1beta, TNFalpha) as well as ET-1 is also increased, indicating the important role of immune reactions in the development of damage to the brain following ischemia. PMID- 19812917 TI - Factors in creepy delayed neuronal death in hippocampus following brain ischemia reperfusion injury with long-term survival. AB - Brain ischemia is well known for its ability to compromise the function of the blood-brain barrier. We assessed blood-brain barrier integrity by examining the leakage of horseradish peroxidase (HRP) and different fragments of amyloid precursor protein from the vascular network into hippocampal parenchyma in rats exposed to brain ischemia with long-term survival. The areas of blood-brain barrier leakage were associated with increased staining of HRP and C-terminal of amyloid precursor protein/beta-amyloid peptide in perivascular space suggesting, respectively, an additional response to ischemia and neuronal death. These results suggest that the events associated with delayed neuronal death in hippocampus compromise blood-brain barrier function. Additionally, these data suggest that the leakage of cytotoxic amyloid precursor protein parts in the CA1 and other sectors of hippocampus may play a role in the development of creepy delayed neuronal death after the ischemia-reperfusion injury. These findings also suggest that the blood-brain barrier vessels along the hippocampal fissure especially in the medial part of the hippocampus are more vulnerable to ischemic episodes than those in other hippocampal areas. PMID- 19812918 TI - Endogenous pituitary adenylate cyclase activating polypeptide is involved in suppression of edema in the ischemic brain. AB - Pituitary adenylate cyclase activating polypeptide is a pleiotropic neuropeptide. We previously showed that heterozygous PACAP gene knockout (PACAP(+/-)) mice had larger infarct volumes and worse neurological scores after middle cerebral artery occlusion (MCAO). However, the relationship between endogenous PACAP levels and edema in the ischemic brain has not yet been evaluated. In this study, the formation of edema in the ischemic brain as well as cerebral blood flow was compared between PACAP(+/-) and wild-type (PACAP(+/+)) mice. The amount of brain edema was calculated by subtracting the contralateral volume from the ipsilateral volume 24 h after permanent MCAO. PACAP(+/-) mice showed significantly greater brain edema than PACAP(+/+) mice. To investigate the effects of endogenous PACAP on blood flow during ischemia, cerebral blood flow in the ipsilateral and the contralateral cortices was compared between PACAP(+/-) and PACAP(+/+) mice for 25 min after ischemia. With a two-dimensional laser Doppler perfusion imaging system, the blood flow in the ipsilateral and contralateral cortices was shown to be similar in PACAP(+/-) and PACAP(+/+) mice during ischemia. These results suggest that endogenous PACAP suppresses the formation of edema in the ischemic brain. PMID- 19812919 TI - Reduced matrix metalloproteinase-9 activity and cell death after global ischemia in the brain preconditioned with hyperbaric oxygen. AB - Matrix metalloproteinase-9 (MMP-9) plays a deleterious role in cell death after global cerebral ischemia. Preconditioning with hyperbaric oxygen (HBO-PC) reduces neuronal damage in the post-ischemic brain; however, its effect on ischemia induced increase in MMP-9 activity and expression remains unexplored.We investigated effects of HBO-PC on alterations in MMP-9 activity/tissue expression accompanying neuronal death after transient global cerebral ischemia.Male SD rats (300-350 g), were allocated either to non-ischemic (naive control or sham operated) or ischemic (four-vessel occlusion, 4VO; 10 min) groups that were HBO preconditioned (2.5 ATA, 1 h daily for 5 days; the last session 24 h before ischemia) or not. Neurobehavioral deficits were assessed prior to collection of brain tissue for gel zymography (MMP-9) and histology (MMP-9 immunofluorescence, TUNEL) at 0 (without ischemia), 6, 24, 72 h and 7 days after 4VO.Both, MMP-9 levels and cell death increased in the hippocampus at 72 h after 4VO. HBO-PC suppressed postischemic MMP-9 activity and CA1 cell damage, and improved functional performance. The increase in MMP-9 immunoreactivity in the brain was also detected after HBO-PC alone. HBO-PC suppresses MMP-9 activity and expression in the postischemic hippocampus. The mechanism of HBO preconditioning may depend on the induction of MMP-9 in the preischemic phase and may be in part mediated by exhaustion of MMP-9 stores in cerebral tissues. PMID- 19812920 TI - Radiation exposure prior to ischemia decreases lesion volume, brain edema and cell death. AB - PURPOSE: To investigate the neuronal response to ischemic injury following exposure to whole brain proton irradiation. METHODS: Brain only proton irradiation (8 Gy, 250 MeV) was performed ten days prior to middle cerebral artery occlusion (MCAO) in 1 year old male Sprague Dawley rats. MCAO was induced in two animal groups: proton irradiated (MCAO + Rad) and MCAO only. Magnetic resonance imaging (MRI) and quantitative analysis were performed prior to and 2 days after irradiation, and then 2, 14 and 28 days after MCAO. After the last imaging time point animals were sacrificed and TUNEL staining was performed on 4% paraformaldehyde - fixed brain sections. RESULTS: Neuroimaging demonstrated a reduction in ischemic lesion volume in the MCAO + Rad group compared with MCAO alone. Neurological deficits did not differ between ischemia groups. Interestingly, there was a 34% decrease in the number of TUNEL-positive cells in MCAO + Rad brains compared to MCAO alone. CONCLUSION: Our results suggest that radiation treatment reduces brain edema, ischemic lesion volume and peri-ischemic apoptosis. The underlying mechanisms are currently unknown and additional studies will elucidate the significance of these results. PMID- 19812921 TI - Effects of VEGF administration or neutralization on the BBB of developing rat brain. AB - We investigated the effects of exogenous Vascular Endothelial Growth Factor VEGF combined with an enriched environment on BBB integrity after a minimal trauma induced during the first days of the critical visual period in rats, when peak levels of endogenous VEGF secretion are reached. VEGF was administered using osmotic mini-pumps placed in middle cortical layers of P18 Long-Evansrats. Tissue changes were evaluated using conventional histology. BBB integrity was shown by immunohistochemistry techniques for EBA and GluT-1. Mini-pump implantation produced a wider cavity in anti-VEGF infused rats. In VEGF-infused rats there was a damaged region around the cannula that was smaller in rats raised in an enriched environment (EE). The administration of VEGF induced a high concentration of plasma proteins in the neuropil around the point of cannula placement and a high inflammatory reaction. VEGF-infused rats raised in an EE showed a lower degree of extravasation and better tissue preservation. Anti-VEGF administration produced a lower protein expression profile and more widespread deterioration of tissue. Double immunofluorescence for EBA and GluT-1 showed that the administration of VEGF preserves the tissue, which remains present but not fully functional. In contrast, a combination of VEGF administration and an EE partially protects the functionally damaged tissue with a higher preservation of BBB integrity. PMID- 19812922 TI - Alterations in blood-brain barrier function and brain pathology by morphine in the rat. Neuroprotective effects of antioxidant H-290/51. AB - The possibility that stress associated with morphine administration or withdrawal will influence the blood-brain barrier (BBB) dysfunction, brain edema formation and brain pathology was examined in a rat model. Repeated daily administration of morphine (10 mg/kg, i.p.) resulted in drug dependence in rats on the 6th day and onwards. The BBB permeability to Evans blue albumin (EBA) and radioiodine ([131])Iodine did not alter during morphine dependence up to the 12th day. On the other hand, spontaneous withdrawal of morphine on day 1 resulted in profound stress symptoms and breakdown of the BBB to protein tracers in several brain regions. This increase in BBB to protein tracers was most pronounced on the 2nd day of morphine withdrawal. These rats also exhibited marked brain edema and abnormal neuronal and glial cell responses. Pretreatment with an antioxidant H 290/51 markedly attenuated the BBB dysfunction, brain edema formation and brain pathology during morphine withdrawal phases. These observations suggest that psychostimulants and associated oxidative stress are capable to induce brain pathology through modifying the BBB function. PMID- 19812923 TI - Clinical proof of the importance of compliance for hydrocephalus pathophysiology. AB - INTRODUCTION: Recently decreased compliance is discussed as an initially disturbed CSF hydrodynamic parameter in hydrocephalus. MATERIALS AND METHODS: In 180 patients with suspected chronic hydrocephalus we performed a dynamic infusion test, which was not used for shunt indication. Shunt indication was based on long term ICP monitoring. Follow-up was 4.6 +/- 1.8 years. STATISTICS: Spearman-, Kruskal-Wallis-, Wilcoxen-U-test. RESULTS: Resistance to outflow (Rout) and Pressure Volume Index (PVI) alone provide positive predictive values (PPV) and sensitivity, which might be sufficient in daily practice, while negative predictive values (NPV) and specificity are weak. With an intelligent combined algorithm of Rout and PVI at a critical value of ROF of 13 mm Hg/ml x min and 30 ml, a clearly improved outcome prediction is possible. CONCLUSION: These clinical results support recent opinions concerning the meaning of Compliance and Rout in hydrocephalus patho-physiology. PMID- 19812924 TI - An algorithm to assess the rehabilitation potential in patients with chronic hydrocephalus. AB - OBJECTIVE: In clinical practice, it is often necessary to judge the probability of clinical benefit of invasive ancillary tests given to patients with chronic hydrocephalus before they are performed. The aim of the current study was to establish a screening tool for such prediction. MATERIAL AND METHODS: A total of 125 patients with chronic hydrocephalus were assessed using a clinical (HHS) and comorbidity (CMI) grading. These patients were shunted and followed-up for at least one year. The statistical tools of ANOVA, CHI-squared, Spearman, Kuskal Wallis, and Wilcoxen-U-Test were applied. RESULTS: The variables discovered to be of use in prediction were age (p = 0.02), anamnesis duration (p = 0.04), CMI (p < 0.000) and HHS (p = 0.001). A decision tree using solely HHS and CMI was established allowing outcome prediction of sufficient power. Interestingly, in patients who had favourable preconditions, older age impeded not a favourable prognosis. CONCLUSION: With our proposed decision tree, a minimum of data allow a clinician to sufficiently judge whether shunting will be of benefit to a patient, which may help to decide whether invasive ancillary tests are justified. PMID- 19812925 TI - What can be found inside shunt catheters. AB - Obstruction of shunt catheters is one of the main causes of shunt malfunction. The fragments of shunts removed from five patients were examined using scanning electron microscopy with a (SEM) JEOL JSM-6390 LV microscope. Fifteen catheters from the brain ventricle, lumbar space, and peritoneal space were studied. SEM studies showed that the catheters' surfaces were not sufficiently smooth. The inner surface was often covered by a web of collagen fibrils. Aggregates of red and white blood cells, platelets, lymphocytes, mast cells, and macrophages were trapped in the collagen web. Such cellular aggregates formed a coherent, delicate web mainly consisting of ultrastructurally unchanged cellular elements and were well preserved. Other types of aggregates contained completely destroyed cells that appeared to be submerged in thick collagen web fibrils.We also found a few ultrastructural abnormalities among morphologically unchanged cellular elements. The presence of abnormal red cells showing unusual variability in their shape and size including spherocytosis (thickened, spheroid, and crenate red cells), elliptocytosis (elongated, rod-shaped, or tear-drop red cells), the thalassaemic phenotype of red cells (with inclusion of precipitated unstable hemoglobin in the form of Heinz bodies distorting the red cells, leading to their lysis) was a striking finding. Under scanning electron microscopy, we also recognized swollen or crumpled red cells that looked like potato crisps. Aggregation of thickened blood platelets and white cells was observed frequently. Our study confirmed the importance of the smoothness of the inner surface of the catheter. Smoothness can prevent the formation of cell and protein deposits. PMID- 19812926 TI - Glue instead of stitches: a minor change of the operative technique with a serious impact on the shunt infection rate. AB - INTRODUCTION: Shunt infections are still one of the most important complications of shunt surgery. We observed shunt infections coming from wound breakdown due to minimal CSF leakage from subcutaneous CSF accumulation, which is often unavoidable in babies over the borehole, along the fibers of stitches that close the superficial skin. Whether such secondary shunt infections might be overcome by avoiding stitches has been studied. MATERIALS AND METHODS: We examined 90 children experiencing their first shunt insertion between September 1998 and April 2008. We divided the children into two groups. Wound closure was performed with absorbable subcutaneous one-on-one sutures with counter-sunk knots in both groups. In one group, octylcyanoacrylate tissue adhesive was used for the final layer closure of the skin (44 children); in the other group, non-absorbable one on-one single skin sutures were used (46 children). RESULTS: Using the glue, we reduced the wound dehiscence rate from 24% to 2% and the infection rate from 17% to 0%. CONCLUSION: A minimal change of operative technique substantially affects the shunt infection rate due to the extermination of the "wick-effect" along filaments used to close the skin. Furthermore, Dermabond itself has a bactericidal effect. PMID- 19812927 TI - Animal experiments to evaluate complications of foreign materials on silicone with shunt catheters: preliminary results. AB - OBJECTIVE: Use of silicone to manufacture hydrocephalus shunts has been critical for the successful introduction of modern shunt therapy. However reactions to foreign material cause biodegradation, calcification, and massive scarring, and their impact on the still high shunt failure rate might have been undervalued in the past. We established an animal model to simulate the conditions and reactions with the silicone catheter in human patients. METHODS: We implanted catheters from different hydrocephalus shunt manufacturers available on the world market in 12 four-week old Wistar rats. To mimic shearing forces and tensile stress, the tubes were firmly fixed proximally and distally in a growing rat. The catheters remained in the subcutaneous tissue for 1 year before being removed and studied using scanning electron microscopy and histological studies. RESULTS: All of the implanted long catheters showed fractures and calcification on their surfaces, whereas the short fragmented catheters did not. CONCLUSION: The immunological reactions with silicone and the biodegradation of the material can be simulated in this animal model to study details of the pathophysiology of this process. PMID- 19812928 TI - Huge thrombosis as a consequence of VA-shunts. AB - OBJECTIVE: Thrombosis is a rare but serious consequence of VA-shunts. We present two cases of near fatal thrombosis and its successful (but in case 2, atypical) management. RESULTS: Case 1: A 38-year-old woman with VA-shunt suffered from rapidly progressing heart failure and later from progressing underdrainage signs nine years after shunting due to a thrombus on the atrial shunt catheter that occluded >80% of the right atrium. Cardio-surgical removal of thrombus and VA shunt catheter and VP-shunting normalized neurological and cardiological state. Case 2: A 40-year-old woman received a VA-shunt 5 years before she suddenly suffered dyspnea and venous congestion. Secondarily, underdrainage occurred. The underlying huge thrombosis of the superior caval vein could not be excised because the necessary thoracotomy would have interrupted vital venous bypasses along the thoracic wall. Anticoagulants (heparin, cumarin) and ETV relieved all neurological and cardiological symptoms. CONCLUSION: Sudden or unexpected symptoms of cardiac failure in the presence of a VA-shunt must be recognized as serious. Interestingly, despite distal shunt occlusion, underdrainage symptoms might be initially mild. PMID- 19812929 TI - Does idiopathic normal pressure hydrocephalus always mean a poor prognosis? AB - OBJECTIVE: The objective was to assess whether idiopathic normal-pressure hydrocephalus (iNPH) has a worse prognosis than other forms of hydrocephalus, as has been suggested. METHODS: A total of 125 patients with chronic hydrocephalus, 75 of whom suffered from iNPH and the remaining (non-INPH) from sNPH or non communicating hydrocephalus, were shunted using gravitational valves. Clinical state was assessed with our clinical grading (KI) and a co-morbidity index (CMI). Average follow-up was 5.1 +/- 1.6 years. STATISTICS: Spearman, Kruskal-Wallis, ANOVA, chi(2)- and the Wilcoxon U tests at a significance level of pi < 0.05 were used. RESULTS: Shunt responder rates for iNPH and non-iNPH were 72% and 86%, respectively. With shorter anamnesis (< or =1 year) or preoperative KI < 6 points, iNPH patients had a similar or even better outcome than non-iNPH patients with longer anamnesis or a worse KI. Most impressive was the influence of co morbidity: 86% of iNPH patients with a low CMI (< or =3 points) experienced clinical improvement after shunting, which was contrasted by a responder rate of 64% for non-iNPH with worse CMI. CONCLUSION: The diagnosis of iNPH does not by itself mean a worse prognosis, and iNPH patients with favorable preconditions may have a similar or better prognosis than patients with any other kind of hydrocephalus. The worse overall clinical results of iNPH result from late recognition and in most instances worse preconditions. PMID- 19812930 TI - Gravitational shunt complications after a five-year follow-up. AB - INTRODUCTION: Gravitational shunts (G-valves) for ventriculo-peritoneal (VP) shunting have been available since 1996. We analyzed shunt complications in patients with a complete minimum follow-up of 5 years. MATERIAL AND METHODS: Between 1996 and 2002, we implanted 282 VP G-valves in various forms of adult chronic hydrocephalus, of which 130 provided a complete data set with an annual follow-up. Adjustable and non-adjustable G-valves were used: the Miethke Dual Switch valve, the Miethke GAV-valve and a combination of adjustable Codman-Hakim valves with the Miethke Shunt-Assistant. In cases of supposed mechanical shunt failure, the explanted shunts were examined in a bench test. RESULTS: The total complication rate was 21%:3% shunt infections, 3% catheter dislocation/fracture, 5% underdrainage and 9% overdrainage occurred. Half of the overdrainage complications could be managed conservatively. Underdrainage complications resulted from the chosen opening pressure being too high (n = 3), a secondary increase in intraperitoneal pressure (n = 2) or from "real" shunt failure in one case according to bench test results. CONCLUSION: G-valves demonstrate sufficient long-term performance over multiple years, and real shunt-related complications are rare. The frequency of revision due to overdrainage is low (4.5%). PMID- 19812932 TI - Early shunting using the parallel shunt system in hemorrhagic hydrocephalus: in vitro testing of handling, technical complications and clogging rate. AB - In order to minimize the duration of external cerebrospinal fluid (CSF) drainage after hemorrhagic hydrocephalus we are testing a parallel shunt system that can be implanted very early after bleeding. It should be able to tolerate a high load of protein and blood in the CSF. After clearance of the CSF, the valveless arm of the shunt is closed by a percutaneous manipulation and the valve-arm is opened. PMID- 19812931 TI - Is it possible to minimize overdrainage complications with gravitational units in patients with idiopathic normal pressure hydrocephalus? Protocol of the randomized controlled SVASONA Trial (ISRCTN51046698). AB - Overdrainage is a common complication observed after shunting patients with idiopathic normal-pressure hydrocephalus (iNPH), with an estimated incidence up to 25%. Gravitational units that counterbalance intracranial pressure changes were developed to overcome this problem. We will set out to investigate whether the combination of a programmable valve and a gravitational unit (proGAV, Aesculap/Miethke, Germany) is capable of reducing the incidence of overdrainage and improving patient-centered outcomes compared to a conventional programmable valve (Medos-Codman, Johnson & Johnson, Germany). SVASONA is a pragmatic randomized controlled trial conducted at seven centers in Germany. Patients with a high probability of iNPH (based on clinical signs and symptoms, lumbar infusion and/or tap test, cranial computed tomography [CCT]) and no contraindications for surgical drainage will randomly be assigned to receive (1) a shunt assistant valve (proGAV) or (2) a conventional, programmable shunt valve (programmable Medos-Codman).We will test the primary hypothesis that the experimental device reduces the rate of overdrainage from 25% to 10%. As secondary analyses, we will measure iNPH-specific outcomes (i.e., the Black grading scale and the NPH Recovery Rate), generic quality of life (Short Form 36), and complications and serious adverse events (SAE). One planned interim analysis for safety and efficacy will be performed halfway through the study. To detect the hypothesized difference in the incidence of overdrainage with a type I error of 5% and a type II error of 20%, correcting for multiple testing and an anticipated dropout rate of 10%, 200 patients will be enrolled.The presented trial is currently recruiting patients, with the first results predicted to be available in late 2008. PMID- 19812933 TI - Intracranial irregularities beside hydrocephalus in H-Tx rats. AB - INTRODUCTION: It has been well documented that up to 70% of H-Tx rats' offspring suffer from severe hydrocephalus, which can be fatal if it remains untreated. Some offspring also have non-fatal moderate hydrocephalus allowing a normal life expectancy. The objective of this study was finding other morphological intracranial abnormalities that are not directly related to hydrocephalus. METHOD: An MRT for small animals (Bruker, Biospec, Erlangen Germany) with a 2.4 T magnet at 100 MHz has been used to study 98 apparently non-hydrocephalic H-Tx rats. T2-weighted 2D-RARE, T2-weighted 3D-Turbo-RARE sequence and T1-weighted 3D gradient-echo sequences were used. RESULTS: Apart from 36% of animals with moderate or mild hydrocephalus, we found one animal with a cystic cerebellar malformation similar to an arachnoid cyst with minimal space occupying effects. Nine rats had a mild or moderate-sized unilateral enlargement of one lateral ventricle, but a causative occlusion of the Foramen of Monroe could not be verified. Finally, one animal with huge hydrocephalus had a midline cystic malformation between both cerebral hemispheres. CONCLUSION: Aside from the well documented hydrocephalus, H-Tx rats may develop other intracranial malformations that have not yet been documented in the literature. PMID- 19812934 TI - Co-morbidity as a predictor of outcome in patients with idiopathic normal pressure hydrocephalus. AB - INTRODUCTION: A critical question in the diagnosis and treatment of idiopathic normal-pressure hydrocephalus (iNPH) is which preoperative factors can most reliably predict outcomes following shunt insertion. The number and type of co morbidities are increasingly being viewed as important predictive indicators. MATERIALS AND METHODS: Between 1997 and 2005, 100 patients were implanted with a gravitational ventriculo-peritoneal shunt as a treatment for iNPH. All coincident disease processes were recorded. Eighty-two of these patients underwent follow-up of 2 years post-operation. The results of this prospective follow-up examination (Kiefer Score, NPH Recovery Rate) were compared with the preoperative Co morbidity Index (CMI). RESULTS: Of the patients with a CMI score of 0-1 (n = 18), 67% experienced an excellent outcome, 28% a good outcome, and 5% and 0% fair and poor outcomes, respectively. A CMI score of 2-3 was associated with markedly poorer outcomes (n = 33); 42% excellent, 30% good, 18% fair and 10% poor. A score of 4-5 was related to 14% excellent, 27% good, 23% fair and 36% poor outcomes (n = 22). Remarkably, a few patients scoring between 6 and 8 on the CMI scale experienced a favorable outcome. The outcomes for this latter group were 0% excellent, 10% good, 45% fair and 45% poor (n = 9). CONCLUSION: Co-morbidity is a statistically significant predictor of the quality of clinical outcome for patients with iNPH undergoing shunt therapy. PMID- 19812935 TI - Effects of magnesium sulfate infusion on cerebral perfusion in patients after aneurysmal SAH. AB - BACKGROUND: A meta-analysis of current data suggests that magnesium sulfate infusion improves the outcome after aneurysmal subarachnoid hemorrhage through a reduction in delayed ischemic neurological deficit. Two multi-center randomized controlled trials are currently underway to investigate this hypothesis. The possible pharmacological basis of this hypothesis includes neuroprotection and vasodilatation. We aim to investigate the cerebral hemodynamic effects of magnesium sulfate infusion in aneurysmal subarachnoid hemorrhage patients. METHOD: A total of 12 patients who had experienced aneurysmal subarachnoid hemorrhage were randomized to magnesium sulfate infusion (n = 6) or placebo infusion (n = 6) for 14 days. Each patient had two perfusion MRIs performed, one in the first week after subarachnoid hemorrhage and one in the second week after subarachnoid hemorrhage. FINDINGS: Age, sex, and Fisher CT grade were not different between the two groups. All but one patient were of WFNS Grade I to II on presentation. There was no increase in rCBV, rCBF and MTT between the two perfusion scans within the same group or between the two groups. CONCLUSION: Magnesium sulfate infusion, in the dosage of current clinical trials, did not increase cerebral blood volume and cerebral blood flow, as postulated by dilation of small vessels and/or collateral pathways. PMID- 19812936 TI - Timing of serum active MMP-9 and MMP-2 levels in acute and subacute phases after spontaneous intracerebral hemorrhage. AB - Serum active matrix metalloproteinase (MMP)-9 and -2 levels and their tissue inhibitors TIMP-1 and -2 were measured in 28 patients with spontaneous intracerebral hemorrhage (SICH) at 24 h, 48 h and 7 days after bleeding. Perihematomal edema volume was calculated on non-enhanced computed tomography scans by using the formula AxBxC/2 at the same time points. Mean levels of serum active MMP-9 and MMP-2, as well as perihematomal edema volume, were significantly different over time (p < 0.0001). In comparison to values observed at 24 h, serum active MMP-9 mean concentrations increased at 48 h and reached their peak at 7 days, serum active MMP-2 mean levels progressively declined at 48 h and at 7 days, whereas perihematomal edema volume increased at 48 h and at 7 days. Perihematomal edema volume was positively correlated with active MMP-9 and MMP-2 at 24 h (p < 0.02 and p < 0.05, respectively) and with active MMP-9 at 48 h (p < 0.05), but was inversely correlated with active MMP-2 at 7 days (p < 0.02). These findings suggest a different involvement of active MMP-9 and MMP-2 in perihematomal-associated inflammatory response occurring in the transition from acute to subacute phases after SICH. PMID- 19812937 TI - Timing of serum soluble HLA-G levels in acute and subacute phases after spontaneous intracerebral hemorrhage. AB - Serum levels of sHLA-G (sHLA-G1/HLA-G5) antigens and their soluble isoforms, sHLA G1 and HLA-G5, were measured by ELISA in 22 patients with spontaneous intracerebral hemorrhage (SICH) at 24 h, 48 h and 7 days after bleeding. The perihematomal edema volume was calculated on non-enhanced computed tomography scans using the formula AxBxC/2 at the same time points. The mean serum concentrations of sHLA-G1/HLA-G5 and sHLA-G1 as well as the perihematomal edema volume changed significantly over time (p < 0.0001, p < 0.001 and p < 0.0001, respectively), whereas no statistical differences were found in serum HLA-G5 concentrations over the course of the experiment. In comparison to the values found at 24 h, sHLA-G1/HLA-G5 and sHLA-G1 increased at 48 h and then decreased at 7 days, whereas the perihematomal edema volume was more elevated at 48 h and, to a lesser extent, at 7 days. A positive correlation was detected between mean serum sHLA-G1/HLA-G5 and sHLA-G1 levels and perihematomal edema volume at 24 h (p < 0.02) and at 48 h (p < 0.01). Our results may indicate a role for sHLA-G in inflammatory mechanisms related to SICH, where these proteins probably act as anti-inflammatory molecules and are predominantly produced as the sHLA-G1 isoform. PMID- 19812938 TI - Minocycline attenuates brain edema, brain atrophy and neurological deficits after intracerebral hemorrhage. AB - Evidence suggests that microglia activation contributes to brain injury after intracerebral hemorrhage (ICH). The present study aimed to determine if minocycline, an inhibitor of microglia activation, can reduce brain edema, brain atrophy and neurological deficits after ICH.Male Sprague-Dawley rats received an infusion of 100-microL autologous whole blood into the right basal ganglia. Rats received minocycline or vehicle treatment. There were two sets of experiments in this study. In the first set of experiments, the effects of minocycline on ICH induced brain edema were examined at day 3. In the second set, behavioral tests were performed at days 1, 3, 7, 14 and 28. Rats were killed at day 28 for brain atrophy measurement (caudate and lateral ventricle size).Minocycline reduced perihematomal brain edema in the ipsilateral basal ganglia (78.8 +/- 0.4 vs. 80.9 +/- 1.1% in the vehicle-treated group, p < 0.01). Minocycline also improved functional outcome. In addition, minocycline reduced brain tissue loss in the ipsilateral caudate (p < 0.01) and ventricular enlargement (p < 0.05).In conclusion, minocycline attenuates ICH-induced brain edema formation, neurological deficits and brain atrophy in rats suggesting an important role of microglia in ICH-related brain injury. PMID- 19812939 TI - Myocardial dysfunction in subarachnoid hemorrhage: prognostication by echo cardiography and cardiac enzymes. A prospective study. AB - BACKGROUND: The myocardial dysfunction in nontraumatic sub-arachnoid hemorrhage (SAH) is not well understood. Borderline elevations of cardiac biomarkers, electrocardiographic repolarization abnormalities and systolic dysfunction have been reported but the clinical significance of these abnormalities is uncertain. METHODS: Patients without history of cardiac disease were prospectively evaluated for cardiac dysfunction. Myocardial regional wall motion and left ventricular ejection fraction (LVEF) were serially studied by transthoracic echocardiogram along with cardiac enzymes. RESULTS: The mean age of the study population was 53 years. Majority of the patients had aneurysms (N = 38). The mean LVEF was 55 +/- 15%. Eight patients had evidence of WMA, mostly global hypokinesia (63%). The mean LVEF of patients with WMA was significantly lower compared to those without WMA (p < 0.001) at day 0. Systolic function recovered in 25% of these patients. The mean value of troponin was significantly higher in those with WMA (p < 0.001) and mean GCS upon admission was significantly lower (p < 0.001). On multivariate analysis, WMA were associated with poor GCS (p < 0.01) and increased hospitalization (P < 0.01). CONCLUSIONS: WMA with systolic dysfunction occurred in 20% of patients and recovered within 3 days in 25%. Patients with evidence of WMA had a significant myocardial dysfunction, higher troponin levels and poor GCS. PMID- 19812940 TI - The effects of tetrahydrobiopterin on intracerebral hemorrhage-induced brain injury in mice. AB - Tetrahydrobiopterin (BH4) is an essential cofactor for nitric oxide synthase (NOS) and is presently used clinically to treat forms of phenylketonuria. BH4 has been reported to restrain superoxide generation of NOS and chemically reduce superoxide. However, there has been no report concerning the effects of BH4 in intracerebral hemorrhage (ICH). In the present study, we investigated the neuroprotective effect of BH4 against ICH-induced brain injury in a mouse model.A total of 26 male CD1 mice (31-39 g) were divided into sham, ICH-vehicle, and ICH treated with BH4 groups (n = 8 in each group). ICH was induced by collagenase injection into the right basal ganglia. BH4 (20 mg/kg) was administrated intraperitoneally at 1 h after ICH. The effect of BH4 was measured by neurological score and brain water content at 24 h after ICH.Our data demonstrates that ICH caused significant neurological deficit that is associated with brain edema. Treatment with BH4 did not reduce brain edema and neurological deficits at 24 h after ICH in mice. Further study is required to investigate the long-term effect of BH4 in ICH-induced brain injury. PMID- 19812941 TI - Estrogen reduces iron-mediated brain edema and neuronal death. AB - Our previous studies found that 17-beta estradiol attenuates edema formation after intracerebral hemorrhage (ICH). As brain iron overload occurs after ICH and contributes to ICH-induced brain injury, the present study examined the effects of estrogen on iron-induced brain injury in vivo and in vitro.There were two sets of experiments in this study. In the first set, male Sprague-Dawley rats were pretreated with 17-beta estradiol or vehicle prior to an intracerebral injection of ferrous iron. Ferrous iron was injected into the right caudate and the rats were killed 24 h later for brain edema measurement. In the second set, primary cultured neurons were pretreated with different doses of 17-beta estradiol or vehicle for 24 h. The cells were then exposed to ferrous iron for 48 h when culture medium was collected for lactate dehydrogenase measurement. Neuronal death was also assessed by live/dead cell assay.Estrogen pretreatment reduced brain water content (p < 0.01) 24 h after iron injection. Estrogen also protected against iron-induced cell death in cultured neurons. Estrogen reduces iron induced brain edema in vivo and neuronal death in vitro suggesting estrogen could be a potential therapeutic agent for ICH. PMID- 19812942 TI - Improving diagnostic value of CT examinations in hyperacute ischemic stroke. AB - This paper presents a computer assisted support of ischemic stroke diagnosis based on nonenhanced CT examinations acquired in the hyperacute phase of stroke. Computational analysis, recognition, and image understanding methods were used for extraction of the subtlest signs of hypodensity in diagnostically important areas. Starting from perception improvement, suggestive and coarse image data visualization was designed as a complement of the standard diagnosis procedure based on CT scan soft-copy review. The proposed method includes an evidence-based description of ischemic conditions and changes, de-skulling and segmenting of unusual areas, the analysis of hypodensity signs across scales and subbands with noise reduction, and hypodensity extraction. Following visualization, forms of empowered hypodensity symptoms localize suggested ischemic areas in source brain image space. Increased visibility of cerebral ischemia for difficult-to-diagnose cases was experimentally noticed and improved diagnostic value of CT was concluded. PMID- 19812943 TI - Pattern recognition methods in (1)H MRS monitoring in vivo of normal appearing cerebellar tissue after treatment of posterior fossa tumors. AB - The objective of this study was to investigate the metabolic responses of normal appearing cerebellar tissue after posterior fossa tumor treatment, and to identify characteristics of the particular treatment method. Moreover, this work examined the metabolic alterations of normal appearing tissue induced by a particular tumor state including resection, stagnation, progression, and recurrence. The studied group consisted of 29 patients treated for posterior fossa tumors. All of them were irradiated with a total dose of 54 Gy at 1.8 Gy/fraction (median values). In addition, 13 underwent chemotherapy, 25 underwent total tumor resection, 18 were tumor-free in control examinations, 5 had a stable disease, and tumor progression or recurrence was observed in 2 and 4 cases, respectively. The 69 spectra, acquired using a MRI/MRS 2T system, were analyzed using Partial Least Squares Discriminant Analysis (PLS-DA) with orthogonal signal correction (OSC) spectral filtering. A significantly elevated spectral region (0.97-1.55 ppm) was observed in patients after total resection in comparison to non-operated subjects. Patients treated with chemotherapy showed an elevated band between 1.15-1.75 and 2.7-3.0 ppm and had decreases in the remaining parts of the spectra. Increases in lactate and decreases in the remaining metabolites were characteristic for the tumor progression/recurrence group. Pattern recognition methods coupled with MRS revealed significant treatment-dependent alterations in normal appearing cerebellar tissue, as well as metabolic changes induced by tumor progression/recurrence. PMID- 19812944 TI - Metabolic changes in rat brain following intracerebroventricular injections of streptozotocin: a model of sporadic Alzheimer's disease. AB - A decrease in cerebral glucose metabolic uptake is an early and characteristic sign of Alzheimer's disease (AD). Streptozotocin (STZ) is a bacterial toxin which damages insulin-producing cells and insulin receptors. Intracerebroventricular (icv) application of STZ in rats has been found to chronically decrease cerebral glucose uptake and produce other effects that bear a resemblance to several other molecular and pathological features of AD. In the present experiments in vivo (1)H MR Spectroscopy with short echo time (3 ms) was used to non-invasively obtain a neurochemical profile of rat brains, 3 weeks and 2 months after double icv injections of STZ or vehicle. Seventeen metabolites were quantified from 27 microL tissue volume which included hippocampus and a part of cerebral cortex, using the LCModel and unsuppressed water signal as an internal reference. Three weeks after icv STZ several metabolites were significantly decreased, the most prominent changes noted in glycerophosphocholine and phosphocholine (-38 +/- 5%), glutathione (-37 +/- 4%), taurine (-30 +/- 19%), glutamate (-26 +/- 14%), phosphocreatine (-23 +/- 15%) and N-acetylaspartate (-16 +/- 6%). On the contrary, the concentration of N-acetylaspartylglutamate was found significantly increased (+38 +/- 18%). After 2 months some of these changes were even more pronounced. We conclude that in vivo (1)H MRS of rat brain following icv STZ injections provides a new input into a better understanding of the critical dependency of neural function and structure on brain glucose consumption, and may be of relevance in further studies of AD pathomechanism. PMID- 19812945 TI - MR spectroscopic evaluation of brain tissue damage after treatment for pediatric brain tumors. AB - PURPOSE: The aim of this study was to evaluate the metabolic profile of uninvolved brain tissue after treatment for pediatric brain tumors. MATERIAL: A group of 24 patients aged 4-18 years was analyzed after combined treatment for brain tumors. In this group, there were nine medulloblastomas, seven low-grade gliomas, three high-grade gliomas, two ependymomas and three children with conservatively treated diffuse brainstem gliomas. METHODS: Short echo-time (TE = 30 ms) point-resolved spectra were acquired using a 2 T clinical scanner (Elscint Prestige). The ratios of signal intensities for N-acetylaspartate (NAA), choline (Cho), myo-inositol (mI), lactate (Lac), and lipids (Lip) were calculated using the creatine (Cr) signal as an internal reference. The spectra were acquired both from the tumor bed and from contralateral uninvolved brain tissue; only control spectra were analyzed. The first examination was made between the third and sixth month after therapy (24 spectra), the second examination occurred 8-12 months after treatment (15 spectra available), and the third was performed approximately 18 months after completion of therapy (eight spectra available). The results were compared using the t-test for dependent samples. RESULTS: At all time points, the metabolite ratios showed alterations indicating brain tissue damage. The most important were the decrease of NAA/Cr and increase of Lac/Cr and Lip/Cr ratios. The mean NAA/Cr values were 0.91, 0.91, and 0.86, respectively, for the three examinations, while the Lac/Cr and Lip/Cr values were 1.66, 2.11, 1.19 and 12.24, 12.05, 5.69, respectively. Interestingly, in children with supratentorial tumors, a significant increase in NAA/Cr value was observed (from 0.82 to 1.11 in the first and second examinations, respectively; p = 0.0487), which may be indicative of neuronal function recovery. CONCLUSIONS: MRS examinations of uninvolved brain tissue indicate long-lasting metabolic disturbances. However, the NAA/Cr ratio increase may be a sign of at least partial recovery of metabolic function of the brain. PMID- 19812946 TI - The incidence of imaging abnormalities after stereotactic radiosurgery for cerebral arteriovenous and cavernous malformations. AB - OBJECTIVE: The aim of the study was to evaluate the incidence of postirradiation imaging changes after stereotactic radiosurgery for arteriovenous malformations (AVM) and cerebral cavernous malformations (CCM). MATERIAL AND METHODS: A group of 85 patients treated for arteriovenous malformations (62 patients, 73%) and cavernomas (23 patients, 27%) between October 2001 and December 2005 was analyzed. All patients were treated with stereotactic radiosurgery with doses ranging from 8-28 Gy. After the irradiation, magnetic resonance imaging (MRI) or computed tomography (CT) was performed at 6 to 12-month intervals to assess the effects of the treatment. The mean follow-up time for the whole group was 27.3 months; AVM group -- 26 months; CCM group -- 30.9 months. All the imaging data were carefully reviewed to identify the radiological symptoms of postradiosurgical damage. T2 or FLAIR hyperintensity, T1-hypointensity and contrast enhancement on MRI and the presence of hypodense areas and contrast enhancement on CT examinations were assessed. RESULTS: Imaging abnormalities were found in 28 (33%) patients. The symptoms of postradiosurgical damage were observed in 21 (33.9%) patients in the AVM group and 7 (30.4%) patients in the CCM group. Radiological symptoms of radiation necrosis associated with neurological deterioration were identified in two patients with cavernomas, while no radiation necrosis was found in the AVM group. Patients in whom radiological signs of focal brain edema or gliosis existed were asymptomatic. CONCLUSIONS: Radiological symptoms of postradiosurgical damage affected about one third of the irradiated patients, typically without any clinical manifestations. Patients irradiated for CCMs seem to be more prone to develop symptomatic postradiosurgical necrosis; this observation, however, requires further investigation. PMID- 19812947 TI - Magnetic resonance spectroscopic evaluation of brain tissue metabolism after irradiation for pediatric brain tumors in long-term survivors: a report of two cases. AB - OBJECTIVE: The aim of our study was to evaluate the metabolic profile of brain tissue of two long-term survivors of childhood brain tumors. MATERIALS: Two males who were 25 and 33 years old at the time of examination and had been irradiated for brain tumors at the age of 17 and 13 years respectively. The first subject had been operated on radically for medulloblastoma and received craniospinal axis irradiation composed of a whole brain radiotherapy with boost to the posterior fossa (total dose (TD) = 59.4 Gy in 33 fractions) and spinal canal irradiation (TD = 30 Gy in 20 fractions) according to the protocol at the time of treatment. The second subject had previously received whole brain irradiation (TD = 45 Gy in 19 fractions) because of inoperable central region tumor of unknown histology. METHODS: Short echo-time (TE = 30 ms) point-resolved spectra were obtained using a 2 T magnet. Ratios of N-acetylaspartate (NAA), choline (Cho), myo-inositol (mI), lactate (Lac) and lipids (Lip) signal intensities were calculated using the creatine (Cr) signal as an internal reference. The spectra were acquired both from the tumor bed area and uninvolved brain tissue in the first subject, and from uninvolved brain areas of frontal and occipital lobes in the second subject. RESULTS: In both cases, MRS examination revealed ratios of NAA/Cr, Cho/Cr and mI/Cr within normal range in most spectra. Nevertheless, a slight elevation of Lac/Cr (2.47 and 1.05) and a more pronounced elevation of Lip/Cr proportions (45.77 and 3.97 respectively, in uninvolved sites) were detected in both patients. CONCLUSIONS: Metabolic parameters correlated with neuronal function (NAA/Cr) and cell membrane metabolites turnover (Cho/Cr) seem to recover to normal values in long-term survivors of brain tumors. Lac/Cr and Lip/Cr proportions could be considered parameters indicating permanent radiation-induced brain damage; however, this proposal requires further investigation. PMID- 19812948 TI - Evaluation of the late effects of CNS prophylactic treatment in childhood acute lymphoblastic leukemia (ALL) using magnetic resonance spectroscopy. AB - PURPOSE: The aim of the study was to evaluate the late changes seen in Magnetic Resonance Spectroscopy (MRS) of the brain in Acute Lymphoblastic Leukemia (ALL) survivors to assess neurotoxicity following prophylactic treatment with cranial irradiation (CRT) and/or intrathecal (ITMTX) and systemic MTX. MATERIALS AND METHODS: The study was performed on two groups of patients. The first group consisted of 30 children who received CRT and ITMTX, and the second group was comprised of 15 children treated only with ITMTX. All patients were ALL survivors treated between 1994 and 2002. Radiotherapy was performed using two opposite fields for a total dose of 18 Gy. The children were examined from 6 to 12 years after treatment. All children underwent a physical and neurological examination and MRI/MRS studies. (1)H-MR spectra were acquired from frontal and occipital regions of the brain. Mean values and standard deviations were calculated for following metabolite ratios: NAA/Cr, Cho/Cr, Cho/NAA, and mI/Cr. RESULTS: Three of the 45 children (11%) presented with white matter changes upon MRI examination. All children with MRI abnormalities received CRT. In 13 (31%) children, changes in (1)H-MRS metabolite ratios were seen. We observed decreased NAA/Cr and Cho/Cr ratios. MR spectroscopy showed a significant reduction (P < .05) of the mean NAA/Cr ratio in children given CRT. CONCLUSION: MRS is a sensitive detector of late metabolic changes after prophylactic treatment for ALL in childhood. It is able to detect metabolic effects of treatment in patients even when no morphologic changes are visible upon MRI. PMID- 19812949 TI - A non-invasive assessment of intracranial volume reserve by measuring cerebrospinal fluid volume with the aid of CT imaging. AB - INTRODUCTION: Monitoring changes in the intracranial volume (ICV) reserve and intracranial pressure (ICP) is one of the key issues in the treatment of intracranial pathologies. The aim of this study is to develop a method of monitoring the ICV reserve by analyzing CSF volume measured using CT in specific regions. MATERIALS AND METHODS: A total of 20 patients with cerebral injury were evaluated. Analysis was performed using imaging software. On selected scans (three at the basal cistern level and three at the pineal level), the following regions were analyzed: total cerebral surface (TC1, TC2) and bilateral ambient cistern (AC) only at the basal cistern level for cerebrospinal fluid (CSF) volume. Results were correlated with patients' Glasgow Coma Scale (GCS) scores. RESULTS: An increase of CSF volume was observed with an improvement in the GCS. From the examined regions, only AC volume showed a statistically significant linear correlation (p < 0.0005) with GCS. Mean AC: 0.021, 0.454, and 0.678 mL CSF/scan in severe (3-8 pts GCS), moderate (9-12 pts GCS), and mild (13-15 pts GCS) TBI groups, respectively. DISCUSSION: Assessment of CSF volume changes in mL CSF/scan can be conducted using CT. Counting voxels corresponding to the CSF eliminates mistakes due to inaccurate region demarcation. The obtained results (AC volume) show a high correlation with patient state. PMID- 19812950 TI - Is neuroradiological imaging sufficient for exclusion of intracranial hypertension in children? Intracranial hypertension syndrome without evident radiological symptoms. AB - There are still many important questions about algorithms and clinical scenarios in the context of children with clinical intracranial hypertension symptoms (IHS) without radiological findings. Such conditions could appear in different clinical situations, including slit ventricle syndrome, overdrainage syndrome, normal volume hydrocephalus, or idiopathic intracranial hypertension. Many articles have defined specific treatment strategies for various forms of IHS, including ventriculoperitoneal shunting, medication for shunt-related migraine, steroids, and valve upgrades with antisiphoning devices or programmable systems. This study is an attempt to define the proper diagnostic procedures and treatment options for patients with various forms of IHS without evident neuroradiological findings. The authors discuss possible pathological mechanisms leading to IHS in the pediatric population. The authors present six children treated in their center. All of the children presented clinical manifestation of intracranial hypertension without evident neuroradiological findings in CT and/or MRI examinations. In three cases, the final diagnosis was slit ventricle syndrome; in two cases, normal volume hydrocephalus; in another case, idiopathic intracranial hypertension. The treatment options included short-term steroid (dexamethasone) administration and ventriculoperitoneal shunting using programmable systems. In one case of idiopathic intracranial hypertension, ICP monitoring was also performed. The authors discuss possible diagnostic and treatment strategies for the aforementioned cases. There are still many controversies about management of children with clinical symptoms of intracranial hypertension that are not confirmed in neuroimaging. It seems that our understanding of intracranial hypertension in the pediatric population is not nearly as sophisticated or complete as we might have imagined. Ventriculoperitoneal shunting with antisiphoning devices and/or short-term dexamethasone administration seem to be the best treatment options in these cases. PMID- 19812951 TI - Substance P immunoreactivity increases following human traumatic brain injury. AB - Recent experimental evidence suggests that neuropeptides, and in particular substance P (SP), are released following traumatic brain injury (TBI) and may play a significant role in the aetiology of cerebral edema and increased intracranial pressure. Whether SP may play a similar role in clinical TBI remains unknown and was investigated in the current study. Archival post-mortem material was selected from patients who had sustained TBI, had died and had undergone post mortem and detailed neuropathological examination (n = 13). A second cohort of patients who had died, but who showed no neuropathological abnormality (n = 10), served as case controls. Changes in SP immunoreactivity were examined in the cerebral cortex directly beneath the subdural haematoma in 7 TBI cases and in proximity to contusions in the other 6 cases. Increased SP perivascular immunoreactivity was observed after TBI in 10/13 cases, cortical neurones in 12/13 and astrocytes in 10/13 cases. Perivascular axonal injury was observed by amyloid precursor protein (APP) immunoreactivity in 6/13 TBI cases. Co localization of SP and APP in a small subset of perivascular fibres suggests perivascular axonal injury could be a mechanism of release of this neuropeptide. The abundance of SP fibres around the human cerebral microvasculature, particularly post capillary venules, together with the changes observed following TBI in perivascular axons, cortical neurones and astrocytes suggest a potentially important role for substance P in neurogenic inflammation following human TBI. PMID- 19812952 TI - Hyperbaric oxygen preconditioning reduces postoperative brain edema and improves neurological outcomes after surgical brain injury. AB - The present study was designed to examine if hyperbaric oxygen preconditioning (HBO-PC) is neuroprotective in a mouse model of surgical brain injury (SBI). C57BL mice were administered 100% oxygen for 1 h at 2.5 ATA for 5 consecutive days and subjected to SBI on the following day. The HBO-PC + SBI animals were compared to sham and normoxia + SBI groups for brain water content in different brain regions at 24 and 72 h after surgery. Blood-brain barrier (BBB) permeability was evaluated using Evan's blue dye extravasation at 24 h. Neurological assessment of the animals was done by a blinded observer at 24 and 72 h. The results showed that brain water content was significantly increased in the right (ipsilateral) frontal lobe surrounding the site of resection. This was attenuated by HBO-PC at 24 and 72 h. However, HBO-PC did not have any effect on the increased BBB permeability observed after SBI. Significant neurological deficits were observed after SBI. HBO-PC improved neurological deficits at 72 h on the 21-point sensorimotor scale and at 24 and 72 h on the wire hang and beam balance scoring. In conclusion, HBO-PC attenuates post-operative brain edema and improves neurological outcomes following SBI. PMID- 19812953 TI - Following brain trauma, copeptin, a stable peptide derived from the AVP precusor, does not reflect osmoregulation but correlates with injury severity. AB - The incidence of water and electrolyte disturbances following traumatic brain injury (TBI) is considerable and has been attributed to a dysregulation of the hypothalamic peptide arginine-vasopressin (AVP). Copeptin, the C-terminal part of the AVP prohormone, reflects AVP activity. In 71 TBI patients we measured copeptin in serum by a sandwich immunoassay. Injury severity was assessed by Glasgow Coma Score (GCS) and computed tomography, and recovery by Glasgow Outcome Score (GOS). Neuroendocrine and osmoregulation regulation were examined on day 0, 3 and 7, and 24 months post-injury. Copeptin was highest on admission (40.0 +/- 72.3 pmol/l), stabilized on day 3 and 7 (21.2 +/- 18.3 resp. 20.3 +/- 17.1 pmol/l), and normalized at follow-up (4.2 +/- 1.7 pmol/l). On admission, there was a correlation between serum sodium and urine excretion (p = 0.003), but the correlation got lost on day 3 and 7. Copeptin did not reflect the individual 24 h urine excretion or serum sodium levels indicating an uncoupling of copeptin/AVP release and renal water excretion. High copeptin level on day 3 were correlated with a low GCS (p < 0.001), midline shift (p = 0.019), intracerebral hemorrhage (p = 0.026), SAPS score (p = 0.001), as well as with a low GOS (p = 0.031). Copeptin was significantly decreased following skullbase fracture (p = 0.016).Our data reveal a loss of hypothalamic osmoregulation following TBI. The measurement of Copeptin/AVP release reveals a significant predictive function for the severity of TBI. PMID- 19812954 TI - The effects of selective brain hypothermia and decompressive craniectomy on brain edema after closed head injury in mice. AB - Intractable brain edema remains one of the main causes of death after traumatic brain injury (TBI). Brain hypothermia and decompressive craniectomy have been considered as potential therapies. The goal of our experimental study was to determine if selective hypothermia in combination with craniectomy could modify the development of posttraumatic brain edema. Male CD-1 mice were anesthetized with halothane and randomly assigned into the following groups: sham-operated (n = 5), closed head injury (CHI) alone (n = 5), CHI followed by craniectomy at 1 h post-TBI (n = 5) and CHI + craniectomy and selective hypothermia (focal brain cooling using cryosurgery device) maintained for 5 h (n = 5). Animals were sacrificed at 7 h posttrauma and brains were removed, sagittally dissected and dried. The brain water content of separate hemispheres was calculated from the weight difference before and after drying. In the CHI alone group there was no significant increase in brain water content in both the ipsi- and contralateral hemispheres (80.59 +/- 1% and 78.74 +/- 0.9% in the CHI group vs. 79.31 +/- 0.7% and 79.01 +/- 0.3% in the sham group, respectively). Brain edema was significantly increased ipsilaterally in the trauma + craniectomy group (82.11 +/ 0.6%, p < 0.05), but not in the trauma + craniectomy + hypothermia group (81.52 +/- 1.1%, p > 0.05) as compared to the sham group (79.31 +/- 0.7%). These data suggest that decompressive craniectomy leads to an increase in brain water content after CHI. Additional focal hypothermia may be an effective approach in the treatment of posttraumatic brain edema. PMID- 19812955 TI - Outcome of patients with severe head injury after decompressive craniectomy. AB - Decompressive craniectomy is an operative option for the neurosurgeon in cases of generalized traumatic brain edema. While the outcome of patients after decompressive craniectomy is often poor, we tried to identify predictors of a favorable course of the injury. Therefore, 131 patients who received a decompressive craniectomy at the Unfallkrankenhaus Berlin (ukb) between September 1997 and September 2005 due to severe traumatic brain injury were followed up. Overall outcome was measured using the Glasgow Outcome Scale (GOS). Sixty-three patients (48%) died during their initial hospital stay and another 27 (21%) were discharged in a vegetative state (GOS 2). Thirty-two patients (24%) were discharged with severe disability, while another nine (7%) had moderate disability at discharge. At an average of 49 months after surgery, 75 patients (68%) were either dead or in a vegetative state (GOS 1 and 2). The results stress again that the prognosis after traumatic brain injury (TBI) with decompressive craniectomy (DC) is unfavorable. Age, midline shift, and status of the basal cisterns on cranial computed tomography (cCT) were associated with the long-term outcome. When weighing whether to initiate the last resort intervention of decompressive craniectomy, the predictive factors detailed here should be taken into consideration. PMID- 19812956 TI - Assessing the neurological outcome of traumatic acute subdural hematoma patients with and without primary decompressive craniectomies. AB - BACKGROUND: We have investigated the impact of primary decompressive craniectomies on neurological outcomes after adjusting for other predictive variables. METHOD: We have collected data from trauma patients with acute subdural hematomas in a regional trauma center in Hong Kong over a 4-year period. Patient risk factors were investigated using logistic regression. RESULTS: Out of 464 patients with significant head injuries, 100 patients had acute subdural hematomas and were recruited for analysis. Forty-four percent of the patients achieved favorable neurological outcomes after 6 months. Favorable neurological outcomes at 1 year were related to age, pupil dilatation, and motor GCS scores at the time of admission. In the 34 patients who underwent evacuation of acute subdural hematomas, primary decompressive craniectomy was not associated with favorable neurological outcomes. CONCLUSION: Primary decompressive craniectomy failed to show benefit in terms of neurological outcomes and should be reserved for cases with uncontrolled intra-operative brain swelling. PMID- 19812957 TI - Changes in the blood-CSF barrier in experimental traumatic brain injury. AB - PURPOSE: Elevation of intracranial pressure (ICP) is a major complication of traumatic brain injury (TBI), and cerebrospinal fluid (CSF) volume is a key factor in ICP regulation. Choroidal epithelial cells (CEC) form the blood-CSF barrier and their integrity is essential for controlling CSF production. In the current study, the morphology of the CEC was studied from 5 h to 28 day after TBI in the rat. METHODS: Male Sprague-Dawley rats were subjected to severe TBI using the impact-acceleration model, and the ultrastructure of the CEC was studied using transmission (TEM) and scanning (SEM) electron microscopy. RESULTS: Radical ultrastructural changes were seen by TEM in CEC in injured animals. At 5 h post injury cell swelling and incipient cytoplasmic vacuoles were seen. At 24 h most severe changes were noted with extensive widening of intercellular clefts. At 7 day and 14 day post-injury, increased cytoplasmic electron density was evident. At 21 day, most microvilli had bulbous ends, and at 28 day cytoplasmic vacuoles were numerous with widened intercellular clefts. SEM revealed a continuum of changes in all injured animals and most conspicuous was the heterogeneity of surface features, with most cells showing bulbous and cup-shaped microvilli, burr like processes and pits. Epiplexus cells were hypertrophic and more numerous. CONCLUSION: At 4 weeks after trauma, choroidal epithelial cells continued to show morphological alterations suggesting that brain homeostasis was still not restored. PMID- 19812958 TI - Dynamics of S100B release into serum and cerebrospinal fluid following acute brain injury. AB - High S100B serum levels are considered to reflect brain injury severity. However, the dynamics of S100B passage from the cerebral compartment into the blood remain unclear. We examined the temporal profile of S100B release into the cerebrospinal fluid (CSF) and blood in acute brain injury.In patients treated with ventricular drainage (subarachnoid hemorrhage, SAH, n = 23; traumatic brain injury, TBI, n = 19), we measured S100B levels in the serum and CSF. The Glasgow Coma Score (GCS) was assessed daily. Statistical analysis was performed by the Mann-Whitney rank sum test for group differences and by the Pearson correlation.In normal controls (n = 6), S100B levels in the serum (0.05 +/- 0.01 microg/L) comprised around 10% of the CSF concentration (0.66 +/- 0.08 microg/L). Following brain injury, S100B levels were significantly increased in the serum (p < 0.05 in SAH day 2-5, TBI day 1-8) and excessively increased in the CSF (p < 0.05 in SAH and TBI day 1-10). For the individual patient, there was no consistent correlation between S100B levels in serum or CSF and GCS. We therefore calculated the ratio of S100B serum/CSF. Following brain injury, the S100B passage from the CSF to the blood was significantly impaired. Further, higher ratios were correlated with better neurological function (p = 0.002).Because stimulated active S100B release may serve as a repair mechanism, a higher S100B serum/CSF ratio may contribute to neurological recovery. PMID- 19812959 TI - Ultrastructural and immunochemical studies of glial scar formation in diabetic rats. AB - We explored the rebuilding of the brain parenchyma after surgical injury due to reactive astrogliosis. In the present study, we investigated the initial stages of rebuilding in the perilesional cortex of streptozotocin-induced diabetic rats. Our methods utilized ultrastructural and immunohistochemical studies as well as Western blot analysis of glial fibrillary acidic protein (GFAP) and vimentin. Data was collected at 2 days, 7 days and 2 months following a unilateral sensorimotor cortex lesion. Electron-microscopic studies revealed not only formation of glial scar tissue but also ultrastructural features of death in the elements of neurovascular unit. Immunohistochemical studies, confirmed by Western blot analysis, demonstrated the enhancement of vimentin and GFAP immunoreactivity (IR) in astrocytes located in the perilesion cortical area of the diabetic rats that were operated upon. We suggest that the process of rebuilding brain parenchyma following surgical injury may be disturbed by the induction of astrocytes and the degeneration of astrocytes, as well as by morphological changes within capillaries that are accompanied by the presence of macrophages. PMID- 19812960 TI - Significance of monitoring the initial intracranial pressure on hematoma irrigation with trephination therapy for acute subdural hematomas in critical conditions. AB - Acute subdural hematoma (ASDH) patients presenting in a severe condition tend to have poor outcomes due to the significant brain edema required to maintain the ICP at less than 20-25 mmHg. This study compared the surgical outcomes of 16 critically ill patients with ASDH who underwent hematoma irrigation with trephination therapy (HITT) based on their initial ICP values. The initial mean GCS score upon admission was four. A unilateral dilated pupil was seen in one and bilateral dilated pupils were seen in seven patients. The co-existence of a brain contusion was seen in seven patients, brain swelling was noted in six patients, and both basal cistern effacement and a midline shift greater than 5 mm were observed in all patients. The mean initial ICP value was 45 mmHg (range: 3 to 85 mmHg). Ten patients (62.5%) underwent a rapid external decompression to evacuate the hematoma. By using the Glasgow Outcome Scale upon discharge a score of good recovery (GR) was assigned to two (12.5 %), moderate disability (MD) to four (25.0 %), vegetative state (VS) to two (12.5 %), and death (D) to eight (50.0 %) patients. All six patients who showed an initial ICP greater than 60 mmHg died despite intensive care. Eight patients who showed an initial ICP less than 40 mmHg had a favorable outcome, but two patients deteriorated due to a traumatic cerebrovascular disorder. It seems that the initial ICP monitoring with HITT for ASDH patients in critical condition may be an important factor for predicting both surgical outcome and clinical course. PMID- 19812961 TI - Surgical outcome following a decompressive craniectomy for acute epidural hematoma patients presenting with associated massive brain swelling. AB - Acute epidural hematomas (AEDH) are generally managed with rapid surgical hematoma evacuation and bleeding control. However, the surgical outcome of patients with serious brain edema is poor. This study reviewed the clinical outcome for AEDH patients and evaluated the efficacy of the DC, especially in patients with associated massive brain swelling. Eighty consecutive patients surgically treated with AEDH were retrospectively assessed. The patients were divided into two groups: (a) hematoma evacuation (HE: 46 cases) and (b) HE+ an external decompression (ED: 34 cases). The medical charts, operative findings, radiological findings, and operative notes were reviewed. In the poor outcome group, there were 18 patients (72%), with a GCS score of less than 8 (severe injury), and 22 patients (88%) who showed pupil abnormalities. Many more patients showed a midline shift, basal cistern effacement, and brain contusion in comparison to the favorable outcome group. In the favorable outcome group, almost all of the patients (98%) showed less than 12 mm of a midline shift. The influential factors may be age, GCS, pupil abnormalities, size, midline shift, basal cistern effacement, coincidence of contusion and swelling. We conclude that an A DC may be effective to manage the AEDH patients with cerebral contusion or massive brain swelling. PMID- 19812962 TI - Prognosis for severe traumatic brain injury patients treated with bilateral decompressive craniectomy. AB - PURPOSE: Decompressive craniectomy for traumatic brain injury patients has been shown to reduce intracranial hypertension, while it often results in increased brain edema and/or contralateral space-occupied hematoma. The purpose of this study was to determine the prognosis of bilateral decompressive craniectomy in severe head injury patients with the development of either bilateral or contralateral lesions after ipsilateral decompressive craniectomy. METHODS: Twelve patients underwent bilateral decompressive craniectomy among 217 individuals who had been treated with decompressive craniectomy with dural expansion from September 1995 to August 2006. The following patient data were retrospectively collected: age, neurological status at admission, time between injury and surgical decompression, time between first and second decompression, laboratory and physiological data collected in the intensive care unit, and outcome according to the Glasgow Outcome Scale. RESULTS: Patient outcomes fell into the following categories: good recovery (three patients); mild disability (one patient); severe disability (two patients); persistent vegetative state (one patient); and death (five patients). Patients with good outcomes were younger and had better pupil reactions and neurological statuses on admission. Other factors existing prior to the operation did not directly correlate with outcome. At 24 h post-surgery, the average intercranial pressure (ICP), cerebral perfusion pressure (CPP), glucose level, and lactate level in patients with poor outcomes differed significantly from those of patients with a good prognosis. CONCLUSION: Head injury patients with either bilateral or contralateral lesions have poor prognosis. However, bilateral decompressive craniectomy may be a favorable treatment in certain younger patients with reactive pupils, whose ICP and CPP values are stabilized 24 h post-surgery. PMID- 19812963 TI - Posterior fossa brain tissue injury: developmental, neuropsychological, and neurological consequences of brain tumors in children. AB - The aim of the study was the functional neurodevelopmental assessment of children with posterior fossa tumors, specifically examining whether tumor location in particular cerebellar structures determines particular neuropsychological deficits. The examined group consisted of 34 children treated between 1999 and 2007 at the Division of Pediatric Neurosurgery Silesian University Medical School in Katowice, Poland. Twelve girls and 22 boys between 5 and 21 years of age were examined. The mean age was 12.3 years. There were 21 total and 8 subtotal resections of tumor, and marsupialization was performed in cases of arachnoid cysts. Hydrocephalus in 19 patients was fixed surgically. Histopathological diagnoses of tumors were as follows: 4 medulloblastomas, 8 pilocytic astrocytomas, 6 fibrillary astrocytomas, 1 anaplastic astrocytoma, 2 oligodendrogliomas, 4 anaplastic ependymomas, 1 choroid plexus papilloma, and 5 arachnoid cysts. The children were assessed by age-appropriate tests that examine higher mental functions such as cognitive processes, visual-spatial functions, verbal fluency, planning, sequential memory, and emotions. Additionally, speech examination and tests were performed. The clinical state of all patients was also evaluated, including a full neurological examination. Posterior fossa tumors can disturb normal development of higher mental functions, especially in the development of linguistic and emotional traits. Our study aimed to better understand the functional anatomy of the cerebellum in the context of behavioral changes. Exploration of the consequences of damage to posterior fossa structures may lead to a better understanding of their function in the emotional and cognitive development of children. Moreover, this work may enable the prediction of neurobehavioral disorders and offer appropriate strategies for rehabilitation, qualification, and surgical procedures. PMID- 19812964 TI - The role of eNOS in vascular permeability in ENU-induced gliomas. AB - Brain edema in gliomas is an epiphenomenon related to blood-brain-barrier (BBB) breakdown in which endothelial nitric oxide synthase (eNOS) plays a key role. When induced by vascular endothelial growth factor (VEGF), eNOS synthesizes nitric oxide that increases vascular permeability. We investigated the relationship between eNOS, VEGF and BBB dysfunction in experimental gliomas.Tumors were produced in Sprague-Dawley rats by transplacentary administration of Ethylnitrosourea (ENU). Immunoexpression of eNOS and VEGF(165) was studied to identify locations of vascular permeability. BBB permeability was evaluated using gadolinium and intravital dyes and BBB integrity by endothelial barrier antigen (EBA), glucose transporter-1 (GluT-1) and occludin immunostaining. Low grade gliomas displayed constitutive eNOS expression in endothelial cells and in VEGF-positive astrocytes surrounding vessels. Malignant gliomas overexpressed eNOS in aberrant vessels and displayed numerous adjacent reactive astrocytes positive for VEGF. Huge dilated vessels inside tumors and glomeruloid vessels on the periphery of the tumor showed strong immunopositivity for eNOS and a lack of occludin and EBA staining in several vascular sections. BBB dysfunction on these aberrant vessels caused increased permeability as shown by Gadolinium contrast enhancement and intravital dye extravasation.These findings support the central role of eNOS in intra- and peritumoral edema in ENU induced gliomas. PMID- 19812965 TI - Potential role of CT perfusion parameters in the identification of solitary intra axial brain tumor grading. AB - In this study, neoplastic perfusion abnormalities were investigated by computed tomography perfusion (CTP) scanning in 38 patients with solitary intra-axial brain tumors (19 with high grade gliomas, 7 with low grade gliomas and 12 with brain metastasis). Regional cerebral blood flow (rCBF), cerebral blood volume (rCBV), mean transit time (rMTT) and permeability surface flow (rPSF) levels were measured in two different regions of interest: (1) enhancing or non-enhancing tumor tissue and (2) a mirror area of apparently normal brain tissue located in the contralateral hemisphere. rCBF mean levels were greater in tumoral tissue than in the contralateral area for high-grade gliomas (p < 0.02). rCBV and rPSF mean values were higher in tumoral tissue than in the contralateral area for high grade gliomas (p < 0.01 and p < 0.05, respectively) and metastasis (p < 0.05 and p < 0.001, respectively). rCBV mean values of tumoral tissue were greater in high grade than in low-grade gliomas (p < 0.05). rPSF mean levels of tumoral tissue were higher in metastasis than in low-grade gliomas (p < 0.02). These findings indicate that multi-parametric CTP mapping may contribute to differential diagnosis of solitary intra-axial brain tumors. PMID- 19812966 TI - Neuromodulation with pleiotropic and multimodal drugs -- future approaches to treatment of neurological disorders. AB - Neurologists are confronted with an ever-growing amount of new information regarding the intimate processes taking place in both normal and pathological brains. Concepts like neuroprotection, neurotrophicity, or anoikis and their clinical utility may be of dazzling complexity. This paper briefly reviews some of the mechanisms involved in the pathogenesis of neurological diseases and current therapies. Since it is becoming more and more clear that using neuroprotective molecule with only one mechanism of action in disease treatment is a utopist idea, the research and use of multimodal drugs should be encouraged.It is not easy to find good therapeutic approaches to neurological disorders, especially if we do not have a deep understanding of all the basic endogenous biological processes, pathophysiological processes, and the links between them. PMID- 19812967 TI - Selected combination of neurotrophins potentiate neuroprotection and functional recovery following spinal cord injury in the rat. AB - The possibility that a combination of neurotrophins induces long-lasting neuroprotection of the cord following spinal cord injury (SCI) was examined in a rat model. The SCI was performed by making a unilateral incision into the right dorsal horn of the T10-11 segments and the animals were allowed to survive 5 h after trauma. Different combination of neurotrophins, i.e., BDNF in combination with GDNF, NT-3, or NGF was applied topically over the traumatized spinal cord and motor dysfunction, blood-spinal cord barrier (BSCB) breakdown, edema formation, and cell injury were examined. Topical application of BDNF in combination with GDNF and NGF 30 min (but not 60 or 90 min) at high concentrations (0.5 microg each) after SCI significantly improved motor function and reduced BSCB breakdown, edema formation, and cell injury at 5 h. However, concurrent application of BDNF, IGF-1, and GDNF (but not with NT-3 or NGF) even 60 or 90 min after trauma induced a significant reduction in motor dysfunction and spinal cord pathology. These observations suggest that a combination of neurotrophins may have added therapeutic value in the treatment of SCI, not reported earlier. PMID- 19812968 TI - Antibodies to dynorphin a (1-17) attenuate closed head injury induced blood-brain barrier disruption, brain edema formation and brain pathology in the rat. AB - The potential neuroprotective efficacy of dynorphin A antiserum on BBB dysfunction, edema formation and brain pathology was examined in a closed head injury (CHI) model in the rat. The CHI was produced by an impact of 0.224 N on the right parietal bone under anesthesia by dropping a weight of 114.6 g on the skull from a height of 20 cm through a guide tube. This concussive brain injury resulted in profound BBB disruption as evidenced by leakage of Evans blue and radioiodine in the brain. Edema formation and swelling at 5 h were most pronounced in the contralateral cerebral hemisphere. Pretreatment with dynorphin A antiserum (1:20, monoclonal) infused into the left lateral cerebral ventricle (30 microL in PBS) either 30 min before or 30 min after CHI significantly attenuated BBB dysfunction, brain edema formation, volume swelling and brain pathology. However, no reduction in brain edema, BBB permeability or improved brain pathology was seen when the antiserum was given 60 min post-CHI. These observations are the first to suggest that antiserum to dynorphin when administered into the CSF during early phase of CHI is neuroprotective. Our work further indicates that dynorphin is actively involved in the cellular and molecular mechanisms of edema formation and BBB breakdown in CHI. PMID- 19812969 TI - Serine protease inhibitor attenuates intracerebral hemorrhage-induced brain injury and edema formation in rat. AB - Our previous studies have demonstrated that thrombin plays an important role in intracerebral hemorrhage (ICH)-induced brain injury and edema formation. We, therefore, examined whether nafamostat mesilate (FUT), a serine protease inhibitor, can reduce ICH-induced brain injury. Anesthetized male Sprague-Dawley rats received an infusion of autologous whole blood (100 microL), thrombin (5U/50 microL) or type VII collagenase (0.4 U/2 microL) into the right basal ganglia, the three ICH models used in the present study. FUT (10 mg/kg) or vehicle was administered intraperitoneally 6 h after ICH (or immediately after thrombin infusion) and then at 12-h intervals (six treatments in total, n = 5 in each group). All rats were sacrificed 72 h later. We also examined whether FUT promotes rebleeding in a model in which ICH was induced by intracerebral injection of collagenase. Systemic administration of FUT starting 6 h after ICH reduced brain water content in the ipsilateral basal ganglia 72 h after ICH compared with vehicle. FUT attenuated ICH-induced changes in 8-OHdG and thrombin reduced brain edema. FUT did not increase collagenase-induced hematoma volume. FUT attenuates ICH-induced brain edema and DNA injury suggesting that serine protease inhibitor may be potential therapeutic agent for ICH. PMID- 19812970 TI - State-of-the-art management and monitoring of brain edema and intracranial hypertension in fulminant hepatic failure. A proposed algorithm. AB - AIM: Develop an evidence-based clinical algorithm integrating clinical decision making on intracranial pressure (ICP) monitoring and intracranial hypertension (ICH) management in the setting of fulminant hepatic failure (FHF). MATERIAL AND METHODS: An English-language literature review was conducted using the PubMed database in November 2007. In compiling evidence on current management trends of ICP and FHF, the paired keywords: fulminant hepatic failure and either mannitol, hypertonic saline, hyperventilation, bioartificial liver, hypothermia, indomethacin, thiopental, or propofol were used. In compiling evidence on ICP monitoring in FHF, the terms "intracranial pressure monitoring" and "liver failure" were used. Excluded references were either pertinent to animal research or irrelevant to ICP monitoring and ICH management in the setting of FHF. RESULTS: State-of-the-art management of ICH due to brain edema in FHF includes Class I therapies such as mannitol and hypertonic saline. Bioartificial liver, hypothermia and hyperventilation are supported by Class II evidence. Indomethacin and sedation remain Class III. Monitoring ICP is supported by Class II and III evidence. A clinical algorithm was created based on the existing therapeutic armamentarium and corresponding evidence support. PMID- 19812971 TI - Novel free radical monitoring in patients with neurological emergency diseases. AB - Recent experimental studies have demonstrated that oxidative stress has important roles in various neuronal conditions. Stroke and traumatic brain injury are also related to oxidative stress. However few studies prove the existence of free radicals in humans because they are difficult to measure. We recently developed a technique for free radical and oxidative stress monitoring using the ex vivo electron spin resonance (ESR) spin trapping method in patients with neuroemergency. Blood samples were collected by catheterization of the internal jugular bulb. The alkoxyl radical level was measured by ex vivo ESR spectrometry using 5,5-dimethyl-1-pyrroline-1-oxide (Dojin Chemical, Tokyo, Japan) as a spin trap. Electron spin response detection of the spin adduct was performed at room temperature using a JESREIX X-band spectrometer (JEOL, Tokyo, Japan). As a marker of reactive oxygen species, we also used the diacron-reactive oxygen metabolites test (d-ROM). This method is not invasive for patients, and it is technically easy to execute.Oxidative stress monitoring is useful and may prove valuable for clarifying the pathophysiology of neuroemergency diseases, which has long been hampered by technical difficulties in measuring and monitoring oxidative stress. PMID- 19812972 TI - Cerebrolysin attenuates blood-brain barrier and brain pathology following whole body hyperthermia in the rat. AB - The possibility that Cerebrolysin, a mixture of several neurotrophic factors, has some neuroprotective effects on whole body hyperthermia (WBH) induced breakdown of the blood-brain barrier (BBB), blood-CSF barrier (BCSFB), brain edema formation and neuropathology were examined in a rat model. Rats subjected to a 4 h heat stress at 38 degrees C in a biological oxygen demand (BOD) incubator exhibited profound increases in BBB and BCSFB permeability to Evans blue and radioiodine tracers compared to controls. Hippocampus, caudate nucleus, thalamus and hypothalamus exhibited pronounced increase in water content and brain pathology following 4 h heat stress. Pretreatment with Cerebrolysin (1, 2 or 5 mL/kg i.v.) 24 h before WBH significantly attenuated breakdown of the BBB or BCSFB and brain edema formation. This effect was dose dependent. Interestingly, the cell and tissue injury following WBH in cerebrolysin-treated groups were also considerably reduced. These novel observations suggest that cerebrolysin can attenuate WBH induced BBB and BCSFB damage resulting in neuroprotection. PMID- 19812973 TI - Chronic hypertension aggravates heat stress-induced brain damage: possible neuroprotection by cerebrolysin. AB - Whole body hyperthermia (WBH) aggravates brain edema formation and cell damage in chronic hypertensive rats compared with normotensive animals. In this investigation, we examined the influence of cerebrolysin on WBH-induced edema formation and brain pathology in hypertensive and normotensive rats. Rats subjected to 4 h WBH at 38 degrees C in a biological oxygen demand (BOD) incubator showed breakdown of the blood-brain barrier (BBB), reduced cerebral blood flow (CBF), edema formation and cell injuries in several parts of the brain. These effects were further aggravated in chronic hypertensive rats (two kidney one clip model (2K1C), for 4 weeks) subjected to WBH. Pretreatment with cerebrolysin (5 mL/kg, 24 h and 30 min before heat stress) markedly attenuated the BBB dysfunction and brain pathology in normal animals. However, in hypertensive animals, a high dose of cerebrolysin (10 mL/kg, 24 h and 30 min before heat stress) was needed to attenuate WBH-induced BBB dysfunction and brain pathology. These observations indicate that heat stress could affect differently in normal and hypertensive conditions. Furthermore, our results suggest that patients suffering from various chronic cardiovascular diseases may respond differently to hyperthermia and to neuroprotective drugs, e.g., cerebrolysin not reported earlier. PMID- 19812974 TI - Carbon nanotubes in neuroscience. AB - Carbon nanotubes have electrical, mechanical and chemical properties that make them one of the most promising materials for applications in neuroscience. Single walled and multi-walled carbon nanotubes have been increasingly used as scaffolds for neuronal growth and more recently for neural stem cell growth and differentiation. They are also used in interfaces with neurons, where they can detect neuronal electrical activity and also deliver electrical stimulation to these cells. The emerging picture is that carbon nanotubes do not have obvious adverse effects on mammalian health. Thus in the near future they could be used in brain-machine interfaces. PMID- 19812975 TI - Nanowired-drug delivery enhances neuroprotective efficacy of compounds and reduces spinal cord edema formation and improves functional outcome following spinal cord injury in the rat. AB - The possibility that drugs attached to nanowires enhance their therapeutic efficacy was examined in a rat model of spinal cord injury (SCI). Three Acure compounds AP-173, AP-713 and AP-364 were tagged with TiO(2)-based nanowires (50 60 nm) and applied over the traumatized cord either 5 or 60 min after SCI in rats produced by a longitudinal incision into the right dorsal horn of the T10-11 segments under equithesin anaesthesia. Normal compounds were used for comparison. After 5 h SCI, behavioral outcome, blood-spinal cord barrier (BSCB) permeability, edema formation and cell injury were examined. Topical application of nanowired compound AP-713 (10 microg in 20 microL) when applied either 5 or 60 min after injury markedly attenuated behavioral dysfunction at 2-3 h after SCI and reduces BSCB disruption, edema formation and cord pathology at 5 h compared to other compounds. Whereas normal compounds applied at 5 min after injury (but not after 60 min) had some significant but less beneficial effects compared to their nanowired combinations. On the other hand, nanowires alone did not influence spinal cord pathology or motor function after SCI. Taken together, our results indicate that the nanowired-drug-delivery enhances the neuroprotective efficacy of drugs in SCI and reduces functional outcome compared to normal compounds even applied at a later stage following trauma, not reported earlier. PMID- 19812976 TI - A new antioxidant compound H-290/51 attenuates nanoparticle induced neurotoxicity and enhances neurorepair in hyperthermia. AB - Previous reports from our laboratory show that animals treated with engineered nanoparticles derived from metals for 1 week and subjected to hyperthermia showed enhanced neurotoxicity in terms of blood-brain barrier (BBB) disruption, brain edema formation and cell injury. It appears that nanoparticle induced enhanced oxidative stress leads to increased lipid peroxidation and over-production of hydroxyl radicals are responsible for exacerbation of neurotoxicity in hyperthermia. Therefore, in this investigation, rats (after 1 week administration of Ag or Cu nanoparticles) were treated with a new antioxidant compound H-290/51 (an inhibitor of lipid peroxidation, 50 mg/kg, p.o.) before subjecting them to hyperthermia. One group of nanoparticle treated rat received H-290/51 and were kept at room temperature for comparison. Our results show that H-290/51 significantly attenuated heat stress induced BBB impairment, brain edema formation and neurotoxicity in nanoparticle treated rats. However, no significant diminution of nanoparticle induced BBB breakdown, or neurotoxicity was observed in H-290/51 treated rats kept at room temperature. These observations suggest that nanoparticles aggravate oxidative stress following hyperthermia leading to exacerbation of neurotoxicity through oxidative stress-related mechanisms, not reported earlier. PMID- 19812977 TI - Influence of nanoparticles on blood-brain barrier permeability and brain edema formation in rats. AB - Nanoparticles are small sized (1-100 nm) particles derived from transition metals, silver, copper, aluminum, silicon, carbon and metal oxides that can easily cross the blood-brain barrier (BBB) and/or produce damage to the barrier integrity by altering endothelial cell membrane permeability. However, the influence of nanoparticles on BBB integrity is still not well-known. In this investigation, effect of nanoparticles derived from Ag, Al and Cu (50-60 nm) on BBB permeability in relation to brain edema formation was examined in a rat model. Intravenous (30 mg/kg), intraperitoneal (50 mg/kg) or intracerebral (20 microg in 10 microL) administration of Ag, Cu or Al nanoparticles disrupted the BBB function to Evans blue albumin (EBA) and radioiodine in rats 24 h after administration and induced brain edema formation. The leakage of Evans blue dye was observed largely in the ventral surface of brain and in the proximal frontal cortex. The dorsal surfaces of cerebellum showed mild to moderate EBA staining. These effects were most pronounced in animals that received Ag or Cu nanoparticles compared to Al nanoparticles through intravenous routes. These observations are the first to suggest that nanoparticles can induce brain edema formation by influencing BBB breakdown in vivo. PMID- 19812978 TI - Functional morphology of the gastrointestinal tract. AB - The primary function of the gastrointestinal tract is water, electrolyte, and nutrient transport. To perform this function, the epithelium lining the gastrointestinal tract is in close contact with the gastrointestinal lumen. Because the lumen is connected to the external environment and, depending on the site, has a high bacterial and antigen load, the epithelium must also prevent pathogenic agents within the gastrointestinal lumen from gaining access to internal tissues. This creates a unique challenge for the gastrointestinal tract to balance the requirements of forming a barrier to separate the intestinal lumen from underlying tissue while simultaneously setting up a system for moving water, electrolytes, and nutrients across the barrier. In the face of this, the epithelial cells of the gastrointestinal tract form a selectively permeable barrier that is tightly regulated. In addition, the intestinal mucosa actively participates in host defense by engaging the mucosal immune system. Complex tissue organization and diverse cellular composition are necessary to achieve such a broad range of functions. In this chapter, the structure and function of the gastrointestinal tract and their relevance to infectious diseases are discussed. PMID- 19812979 TI - Vibrio cholerae interactions with the gastrointestinal tract: lessons from animal studies. AB - Vibrio cholerae is a curved Gram-negative rod that causes the diarrheal disease cholera. One hundred and twenty five years of study of V. cholerae microbiology have made this lethal pathogen arguably the most well-understood non-invasive mucosal pathogen. Over the past 25 years, modern molecular techniques have permitted the identification of many genes and cellular processes that are critical for V. cholerae colonization of the gastrointestinal tract. Review of the literature reveals that there are two classes of genes that influence V. cholerae colonization of the suckling mouse intestine, the most commonly used animal model to study V. cholerae pathogenesis. Inactivation of one class of genes results in profound attenuation of V. cholerae intestinal colonization, whereas inactivation of the other class of genes results in only moderate colonization defects. The latter class of genes suggests that V. cholerae may colonize several intestinal niches that impose distinct requirements and biological challenges, thus raising the possibility that there is physiologic heterogeneity among the infecting population. Efficient V. cholerae intestinal colonization and subsequent dissemination to the environment appears to require temporally ordered expression of sets of genes during the course of infection. Key challenges for future investigations of V. cholerae pathogenicity will be to assess the degree of heterogeneity in the infecting population, whether such heterogeneity has functional significance, and if stochastic processes contribute to generation of heterogeneity in vivo. PMID- 19812980 TI - Interaction of Yersinia with the gut: mechanisms of pathogenesis and immune evasion. AB - Yersinia entercolitica and Yersinia pseudotuberculosis are human foodborne pathogens that interact extensively with tissues of the gut and the host's immune system to cause disease. As part of their pathogenic strategies, the Yersinia have evolved numerous ways to invade host tissues, gain essential nutrients, and evade host immunity. Technological advances over the last 10 years have revolutionized our understanding of host-pathogen interactions. The application of these new technologies has also shown that even well-understood pathogens such as the Yersinia have many surprises waiting to be revealed. The complex interaction with the host has made Yersinia a paradigm for understanding bacterial pathogenesis and the host response to invasive bacterial infections. This review examines the mechanisms of immune evasion employed by the Yersinia and highlights recent advances in understanding the host-pathogen interaction. PMID- 19812981 TI - Molecular mechanisms of Salmonella virulence and host resistance. AB - Salmonella species can cause typhoid fever and gastroenteritis in humans and pose a global threat to human health. In order to establish a successful infection, Salmonella utilize a large number of genes encoding a variety of virulence factors. Different animal models of infection have been used to better understand the mechanisms underlying each disease including cattle, rodents, and nematodes. To date, a number of different bacterial virulence factors have been identified using such animal models, most of which are secreted by two type three secretion systems (T3SS) encoded within Salmonella pathogenicity islands (SPI) 1 and 2. These proteins alter various host cell pathways, facilitating the invasion of epithelial cells during infection, as well as the survival and replication of Salmonella inside phagocytic cells. On the other hand, host genetics and resistance also play a role in the susceptibility to Salmonella infection. The natural resistance-associated macrophage protein 1 (Nramp1), for example, is critical for host defense, since mice lacking Nramp1 fail to control bacterial replication and succumb to low doses of S. Typhimurium. In this chapter, we analyze the different pathogen and host factors that play a role in the dynamic interaction between Salmonella and its host and their impact on disease. PMID- 19812982 TI - Virulence mechanisms and persistence strategies of the human gastric pathogen Helicobacter pylori. AB - The human gastric pathogen Helicobacter pylori is able to establish an infection in a hostile environment with virtually no competitors. For this purpose, it has elaborated a set of colonization factors which facilitate both survival under acid exposure, motility and orientation in a highly viscous mucus layer, and adherence to epithelial surfaces. A more intimate interaction with gastric epithelia provides the basis to influence gene expression profiles as well as morphological transitions via signaling cascades or via direct activities of virulence factors. H. pylori is also one of the most genetically diverse of organisms, and variations are not only found in outer membrane adhesins, but also in two major virulence factors, the VacA cytotoxin and the cag pathogenicity island. Both factors are able to target different cell types and different interaction partners to induce a wide range of possible cellular effects. Despite the fact that H. pylori elicits a strong inflammatory response, the immune system fails to clear the infection, suggesting that immune evasion strategies are used. The mechanisms for immune evasion include the induction of a strongly polarized immune response, a modulation of phagocytosis and neutrophil function, and an inhibition of lymphocyte proliferation. Prolonged inflammation and direct action of bacterial factors may lead to impairment of gland function and eventually to carcinogenesis. PMID- 19812983 TI - Listeria as an enteroinvasive gastrointestinal pathogen. AB - The bacterium Listeria monocytogenes is the causative agent of listeriosis, a highly fatal opportunistic foodborne infection. Listeria spp. are isolated from a diversity of environmental sources, including soil, water, effluents, a large variety of foods, and the feces of humans and animals. Recent outbreaks demonstrated that L. monocytogenes can cause gastroenteritis in otherwise healthy individuals and more severe invasive disease in immunocompromised patients. Common symptoms include fever, watery diarrhea, nausea, headache, and pains in joints and muscles. The intestinal tract is the major portal of entry for L. monocytogenes, whereby strains penetrate the mucosal tissue either directly, via invasion of enterocytes, or indirectly, via active penetration of the Peyer's patches. Studies have revealed the strategy taken by the bacteria to overcome changes in oxygen tension, osmolarity, acidity, and the sterilizing effects of bile or antimicrobial peptides to adapt to conditions in the gut. In addition, L. monocytogenes has evolved species-specific strategies for intestinal entry by exploiting the interaction between the internalin protein and its receptor E cadherin, or inducing diarrhea and an inflammatory response via the activity of its hemolytic toxin, listeriolysin. The ability of these bacteria to survive in bile-rich environments, and to induce depletion of sentinel cells such as Paneth cells that monitor the luminal burden of commensal bacteria, suggest strategies that have evolved to promote intestinal survival. Preexisting gastrointestinal disease may be a risk factor for infection of the gastrointestinal tract with L. monocytogenes. Currently, there is enough evidence to warrant consideration of L. monocytogenes as a possible etiology in outbreaks of febrile gastroenteritis, and for further studies to examine the genetic structure of Listeria strains that have a propensity to cause gastrointestinal versus systemic infections. PMID- 19812984 TI - Molecular mechanisms of campylobacter infection. AB - Campylobacter jejuni is the principal bacterial foodborne pathogen. A major challenge still is to identify the virulence strategies exploited by C. jejuni. Recent genomics, proteomics, and metabolomics approaches indicate that C. jejuni displays extensive inter- and intrastrain variation. The diverse behavior enables bacterial adaptation to different environmental conditions and directs interactions with the gut mucosa. Here, we report recent progress in understanding the molecular mechanisms and functional consequences of the phenotype diversity. The results suggest that C. jejuni actively penetrates the intestinal mucus layer, secretes proteins mainly via its flagellar apparatus, is engulfed by intestinal cells, and can disrupt the integrity of the epithelial lining. C. jejuni stimulates the proinflammatory pathway and the production of a large repertoire of cytokines, chemokines, and innate effector molecules. Novel experimental infection models suggest that the activation of the innate immune response is important for the development of intestinal pathology. PMID- 19812985 TI - Shigella infection of intestinal epithelium and circumvention of the host innate defense system. AB - Shigella, Gram-negative bacteria closely related to Escherichia coli, are highly adapted human pathogens that cause bacillary dysentery. Although Shigella have neither adherence factors nor flagella required for attaching or accessing the intestinal epithelium, Shigella are capable of colonizing the intestinal epithelium by exploiting epithelial-cell functions and circumventing the host innate immune response. During Shigella infection, they deliver many numbers of effectors through the type III secretion system into the surrounding space and directly into the host-cell cytoplasm. The effectors play pivotal roles from the onset of bacterial infection through to the establishment of the colonization of the intestinal epithelium, such as bacterial invasion, intracellular survival, subversion of the host immune defense response, and maintenance of the infectious foothold. These examples suggest that Shigella have evolved highly sophisticated infectious and intracellular strategies to establish replicative niches in the intestinal epithelium. PMID- 19812986 TI - [A comparative in vitro study of cell toxicity of clinically used antiseptics]. AB - PURPOSE: Local skin antiseptics are the standard of care for chronic and non healing wounds. However, little is known about their potential toxic properties. This study investigates the impact of three commercially available and widely used antiseptics on vitality and proliferation of human cutaneous cells. MATERIAL AND METHODS: Three antiseptics, Lavasept (PHMB), Octenisept (octenidine) and Betaisodona (PVP-iodine) were tested for their cytotoxic effects towards HaCaT cells, primary human keratinocytes and fibroblasts using MTT assay and BrDU ELISA. RESULTS: Lavasept showed only slight to moderate toxic effects on cellular vitality and proliferation. Ocentisept and Betaisodona induced severe reduction of cell vitality (p<0.05) to 0% surviving fibroblasts at 7.5% (Betaisodona) and 12.5% Octenisept, respectively. Furthermore, poliferative activity was reduced to 0% in keratinocytes at 7.5% concentration of Betaisodona and Ocentisept. CONCLUSION: This study shows that frequently used wound- and skin antiseptics show severe cytotoxic effects towards cutaneous cells. Furthermore, antimicrobial efficacy and toxic properties must be included in the clinical decision process for optimal therapy of chronic wounds. The PHMB solution Lavasept showed best results regarding toxicity in this study. PMID- 19812987 TI - [Current concepts in the therapy of polytraumatized patients]. PMID- 19812988 TI - Differential association of HLA with three subtypes of type 1 diabetes: fulminant, slowly progressive and acute-onset. AB - AIM/HYPOTHESIS: We sought to clarify similarities and differences in the contribution of HLA to genetic susceptibility to three subtypes of type 1 diabetes: acute-onset, fulminant and slowly progressive. METHODS: We genotyped 545 Japanese patients with type 1 diabetes (338 acute-onset, 80 fulminant, 127 slowly progressive) and 396 control participants at HLA-DRB1, -DQB1, -A, -B and C, and at 101 candidate single nucleotide polymorphisms (SNPs) in an 8.5 Mb region of the extended HLA. RESULTS: DRB1*0405-DQB1*0401, DRB1*0802-DQB1*0302 and DRB1*0901-DQB1*0303 were associated with acute-onset type 1 diabetes, with the DRB1*0405-DQB1*0401/DRB1*0802-DQB1*0302 genotype achieving the highest odds ratio of 42.7. DRB1*1501-DQB1*0602 and DRB1*1502-DQB1*0601 were negatively associated with acute-onset type 1 diabetes. A similar tendency was observed for slowly progressive type 1 diabetes. In contrast, only DRB1*0405-DQB1*0401 was associated with fulminant type 1 diabetes, with the DRB1*0405-DQB1*0401/DRB1*0405-DQB1*0401 genotype showing the highest odds ratio of 11.2. DRB1*0802-DQB1*0302, DRB1*0405 DQB1*0401/DRB1*0802-DQB1*0302 and DRB1*1501-DQB1*0602 were not associated with fulminant type 1 diabetes. The association of class I alleles and a panel of SNPs in an extended HLA region with fulminant type 1 diabetes was also different from that seen for the acute-onset and slowly progressive forms. The presence of both one and two susceptible haplotypes conferred susceptibility to slowly progressive type 1 diabetes, whereas the presence of two susceptible haplotypes was required to confer susceptibility to acute-onset and fulminant type 1 diabetes. CONCLUSIONS/INTERPRETATION: These data suggest that HLA associations with fulminant type 1 diabetes are qualitatively different from those with other subtypes of type 1 diabetes, whereas the HLA contribution to slowly progressive type 1 diabetes is qualitatively similar to, but quantitatively different from, that in acute-onset type 1 diabetes. PMID- 19813002 TI - Improvement of hypercortisolism by beta-blocker therapy in subclinical Cushing's syndrome associated with ACTH-independent macronodular adrenocortical hyperplasia. AB - A 61-year-old man with hypertension and diabetes was referred for the evaluation of multiple bilateral adrenal tumors. While Cushingoid features were not apparent, an elevated cortisol level in response to a low-dose dexamethasone suppression test (187.7 nmol/l), an elevated urinary cortisol level (170.9 nmol/day), and a weak response to a cosyntropin-releasing hormone (CRH) provocation test were observed. Furthermore, the serum cortisol level increased in response to a posture test or isoproterenol infusion. Accordingly, the patient was diagnosed as having ACTH-independent macronodular adrenal hyperplasia (AIMAH) with subclinical Cushing's syndrome associated with the aberrant expression of beta-adrenergic receptors. After 2 months of propranolol therapy, the serum cortisol responses to a posture test and isoproterenol infusion, the cortisol level in response to a low-dose dexamethasone suppression test (102.1 nmol/l), and the urinary cortisol level (165.9 nmol/day) all normalized. While the suppression of cortisol secretion was sustained for 24 months, glucose metabolism and adrenal size were unaffected. To our knowledge, this is the first report of AIMAH accompanied by subclinical Cushing's syndrome associated with the aberrant expression of beta-adrenergic receptors. Furthermore, propranolol inhibited cortisol hypersecretion in the present case. Additional cases or controlled studies are needed to determine the potential effect of propranolol on metabolic disorders and adrenal size in patients with AIMAH. PMID- 19813003 TI - Low-dose tryptophan depletion in recovered depressed women induces impairments in autobiographical memory specificity. AB - BACKGROUND: Depressed patients perform poorly on tests of autobiographical memory specificity (AMS); this may have negative consequences for other important cognitive abilities, delays recovery from mood episodes, and, in recovered patients, may mediate vulnerability to future episodes. Although the cognitive mechanisms underlying AMS deficits are beginning to be understood, the neurobiological mechanisms remain unclear. Serotonin is implicated in both depression and long-term memory; therefore, temporary lowering of brain serotonin function via acute tryptophan depletion (ATD) offers a means of studying the role of serotonin in autobiographical memory specificity. MATERIALS AND METHODS: In this study, 24 previously depressed women underwent low-dose ATD or sham depletion and completed tests of initial and delayed memory, recollection- and familiarity-based recognition, and AMS. RESULTS: ATD did not differentially affect state mood. Compared with sham depletion, ATD impaired immediate recall on the Auditory Verbal Learning Test. Although ATD did not differentially impair recollection- and familiarity-based recognition, it did slow recognition of positive words. ATD also reduced autobiographical memory specificity in response to negative cue words. DISCUSSION: The results confirm previous findings that low dose ATD can reinstate depression-congruent biases in cognition without causing depressive mood in vulnerable populations. The ATD-induced reduction in memory specificity suggests that serotonergic dysfunction may mediate depressive deficits in autobiographical memory; the interaction of cognitive and neurobiological vulnerability mechanisms is discussed. PMID- 19813004 TI - Proteomic profiling combining solution-phase isoelectric fractionation with two dimensional gel electrophoresis using narrow-pH-range immobilized pH gradient gels with slightly overlapping pH ranges. AB - This paper describes a simple new approach toward improving resolution of two dimensional (2-D) protein gels used to explore the mammalian proteome. The method employs sample prefractionation using solution-phase isoelectric focusing (IEF) to split the mammalian proteome into well-resolved pools. As crude samples are thus prefractionated by pI range, very-narrow-pH-range 2-D gels can be subsequently employed for protein separation. Using custom pH partition membranes and commercially available immobilized pH gradient (IPG) strips, we maximized the total separation distance and throughput of seven samples obtained by prefractionation. Both protein loading capacity and separation quality were higher than the values obtained by separation of fractionated samples on narrow pH-range 2-D gels; the total effective IEF separation distance was ~82 cm over the pH range pH 3-10. This improved method for analyzing prefractionated samples on narrow-pH-range 2-D gels allows high protein resolution without the use of large gels, resulting in decreased costs and run times. PMID- 19813005 TI - Searching for signals in the noise: metabolomics in chemical ecology. AB - Chemically mediated interactions between organisms influence ecosystem structure, making it crucial for ecologists to understand these interactions. Advances in chemical ecology have often been closely linked to advances in analytical chemistry techniques. One recent development is the use of metabolomics to address questions in chemical ecology. Although metabolomics has much to offer this field, it is not without drawbacks. Here we consider how metabolomics techniques can supplement the traditional bioassay-guided fractionation approach to chemical ecology. We focus on specific examples that illustrate the advantages that metabolomic methods can provide over other methods in order to understand chemically mediated interactions between organisms. PMID- 19813006 TI - Rapid data analysis method for differential mobility cytometry. AB - Differential mobility cytometry (DMC) has recently been established as a powerful method to capture cells and study adhesion processes. DMC uses an oscillation system and cell affinity chromatography to monitor cells as they adhere to a surface. In the past, differential images had to be created individually which limited the throughput of the method. A new method to create differential images is presented. The method involves the subtraction of short movies from each other to create a stack of differential images that can be easily analyzed. In the future, this method will make DMC more accessible and improve throughput. PMID- 19813007 TI - Micro-analytical evidence of origin and degradation of copper pigments found in Bohemian Gothic murals. AB - Correct identification of pigments and all accompanying phases found in colour layers of historical paintings are relevant for searching their origin and pigment preparation pathways and for specification of their further degradation processes. We successfully applied the analytical route combining non-destructive in situ X-ray fluorescence analyses with subsequent laboratory investigation of micro-samples by optical microscopy, scanning electron microscopy/energy dispersive spectroscopy and X-ray powder micro-diffraction (micro-XRD) to obtain efficiently all the data relevant for mineralogical interpretations of the copper pigments origin. Cu salts (carbonates, chlorides, sulphates, etc.) used as pigments exist in a range of polymorphs with similar or identical composition. The efficiency of the micro-XRD for direct identification of such crystal phases present in micro-samples of colour layers was demonstrated in the presented paper. A new, until now unpublished, type of copper pigment--cumengeite, Pb(21)Cu(20)Cl(42)(OH)(40)--used as a blue pigment on a sacral wall painting in the Czech Republic was found by means of micro-XRD. Furthermore, azurite, malachite, paratacamite, atacamite and posnjakite were identified in fragments of colour layers of selected Gothic wall paintings. We found Cu-Zn arsenates indicating the natural origin of azurite and malachite; artificial malachite was distinguishable according to its typical spherulitic crystals. The corrosion of blue azurite to green basic Cu chloride was clearly evidenced on some places exposed to the action of salts and moisture-in a good agreement with the results of laboratory experiments, which also show that oxalic acid accelerates the corrosion of Cu pigments. PMID- 19813008 TI - Investigations of inter- and intraindividual relationships between exposure to oral salmon calcitonin and a surrogate marker of pharmacodynamic efficacy. AB - AIMS: The aims of the study were to investigate interindividual variations in the bioavailability of salmon calcitonin (sCT) following single oral 0.8 mg doses at three different times of the day, and intraindividual variation in sCT bioavailability at each end of a 14-day treatment period. We also investigated correlations between exposure to sCT and levels of the bone resorption biomarker serum C-terminal telopeptide of collagen type I (CTX-I). METHODS: Participants were from two randomized, double-blind, placebo-controlled studies. In study I, healthy postmenopausal women received a single dose of 0.8 mg of oral sCT or placebo at 08:00 (n = 42), 17:00 (n = 20), or at 22:00 (n = 19). In study II, age matched men or postmenopausal women with osteoarthritis received 0.8 mg oral sCT (n = 26) or placebo (n = 23) twice daily for 14 days, with dosing at 08:00 and 17:00. In both studies, drug exposure was assessed by plasma sCT concentrations, and bone resorption by CTX-I levels. RESULTS: The variability in exposure between patients, measured as coefficient of variation (CV), was as follows: 22% for the morning dose, 30% for the predinner dose, and 34% for the evening dose. In study 1, a high degree of correlation was seen between the level of exposure following a single 0.8 mg dose of sCT and suppression of serum CTX-I, with Pearson correlation coefficients of r = -0.74, -0.96, and -0.78, following doses at 08:00, 17:00, and 22:00, respectively. In study II, exposure to sCT varied widely within the same individuals between dosing days 1 and 14, with weak correlations of r = 0.40 and 0.38 at the dose times 08:00 and 17:00, respectively. As expected from this finding, the intraindividual response in serum CTX-I levels was non significantly associated on dosing days 1 and 14 (r = 0.34 and r = 0.27 at dose times 08:00 and 17:00, respectively). CONCLUSION: Increased bioavailability of orally administered 0.8 mg sCT was highly correlated with increased suppression of the bone resorption marker serum CTX-I irrespective of the time of day. However, the high inter- and intraindividual variability in sCT exposure demonstrates the importance of determining the optimum conditions for ensuring the most beneficial sCT uptake. PMID- 19813009 TI - Pectus excavatum imaging: enough but not too much. AB - BACKGROUND: Pectus excavatum (pectus) is a common congenital deformity of the chest wall resulting in a diminished anterior-posterior dimension. Chest CT has become a common study for preoperative assessment. CT evaluation was initially described using a single CT image; it is now common to perform a CT of the entire chest to evaluate pectus. OBJECTIVE: To evaluate the efficacy of chest radiographs compared to chest CT in identifying additional clinically significant abnormalities in the preoperative evaluation of children with pectus. MATERIALS AND METHODS: We reviewed the chest CT scans of 209 children and young adults who had been evaluated for possible surgical repair of pectus. Additional abnormalities were categorized as (1) incidental, (2) potentially significant, and (3) findings that affected the decision to perform surgery. Chest radiographs were reviewed for category 3 findings. RESULTS: Seventy-six scans showed additional abnormalities, five in group 2 and two in group 3. Both group 3 findings, a vascular ring and an acute pneumonia, were identified on chest radiographs. CONCLUSION: Conventional radiographs identified clinically important findings in children and young adults evaluated for pectus surgery. Radiation risks and medical costs might be substantially decreased by obtaining a chest radiograph and using a limited CT technique when a CT scan is ordered for the purpose of obtaining a Haller index. PMID- 19813010 TI - Dysplasia epiphysealis hemimelica: radiographic and magnetic resonance imaging features and clinical outcome of complete and incomplete resection. AB - OBJECTIVES: The objectives of this communication were to discuss radiographic and magnetic resonance (MR) imaging manifestations and clinical outcome after complete and incomplete resection of the mass of dysplasia epiphysealis hemimelica (DEH). MATERIALS AND METHODS: Clinical records, radiographs, and MR images of eight patients with DEH were retrospectively examined. Six patients were treated by complete excision of the lesional mass, and two patients were treated by incomplete resection at our University Hospitals during the period from 1980 to 2006. RESULTS: We found that, unlike in osteochondroma, DEH was radiographically not clearly separable from the underlying or host bone with preserved cortical bone and marrow continuity. The finding in the talus distinguished DEH from (osteochondroma-like) parosteal osteosarcoma, in which a radiolucent demarcation line clearly separated the tumor from the host bone. The DEH mass had a well-defined low to intermediate signal intensity on T1-weighted images and an intermediate to high signal intensity on T2-weighted images, with irregularity of the articular surface. Simple excision was performed in all patients. The excision was complete in six patients and incomplete in two patients whose lesions was juxta-articular in the ankle and articular in the knee, respectively. The residual mass slowly absorbed and vanished, resulting in mild flaring of the affected portion of the epiphysis. No local recurrence or complication was seen in any of the eight patients. CONCLUSIONS: Although the radiographic signs of DEH are characteristic, (osteochondroma-like) parosteal osteosarcoma should be differentiated from DEH when there is a radiolucent separation line between the mass and host bone in the talus. Simple excision was effective in the management of DEH if the deformity was not complicated. Incompletely excised masses resolved and vanished with time. PMID- 19813011 TI - Interlocking nailing without imaging: the challenges of locating distal slots and how to overcome them in SIGN intramedullary nailing. AB - Placement of the distal interlocking screw is the most difficult part in all intramedullary nail interlocking screw systems and the Surgical Implant Generation Network (SIGN) interlocking system is not an exception. SIGN nails are interlocking implants designed with a precision instrumentation set for use in treatment of long bone fractures without an image intensifier. Locating the distal slots of SIGN nails could be challenging for young SIGN surgeons when treating very complex comminuted fractures and in obese patients. This study was stimulated by a patient who presented one year after surgery with knee pain due to a migrating nail because of missed distal screws. A total of 48 patients divided into two groups of 24 were studied retrospectively and prospectively. The retrospective studies revealed that failure to locate distal locking slots in ten antegrade nailing procedures was due to wrong entry point and comminution of the fracture. The challenges encountered led us to innovating methods to overcome the difficulties of placement of distal screws in a prospective study. Application of methods A and B made location of the distal slots easier in the prospective study even though there were more complex comminuted fractures. The methods also reduced the antegrade operation time by 1 hour 11 minutes. We concluded that SIGN nailing could be challenging and frustrating at the early learning stage. Application of the two innovative methods will make distal slot location easier. They will also make SIGN interlocking nailing less difficult for young SIGN surgeons as they journey through the learning curves. PMID- 19813012 TI - Correlation of positioning and clinical results in Oxford UKA. AB - In a prospective clinical study, 59 patients with anteromedial osteoarthritis of the knee (61 knee joints) underwent minimally invasive medial Oxford unicompartmental arthroplasty phase 3. Clinical and radiographic examinations of 56 knees were carried out at five (4-7) years. American Knee Society (AKS) scores improved from mean 45.5 (20-80) points (knee score) and 55 (15-100) points (function score) before surgery to 90 (30-100) points in both scores after surgery. The position of each implant was determined on screened radiographs using an image intensifier. The implant position was analysed according to the Oxford X-ray rating system. We evaluated nine measures, and there was no detectable correlation between implant position and clinical result. However, long-term studies are needed before it is possible to elaborate an evidence-based guideline on positioning. PMID- 19813013 TI - Cell cycle and apoptosis regulatory gene expression in the bone marrow of patients with de novo myelodysplastic syndromes (MDS). AB - Deregulation of cell cycle and apoptosis pathways are known contributors to the pathogenesis of myelodysplastic syndromes (MDS). However, the underlying mechanisms are not fully clarified. The aim of our study was to examine mRNA expression levels of cell cycle and apoptosis regulatory genes, as well as the percentage of apoptotic and S phase cells and to correlate the findings with clinical characteristics and prognosis. Sixty patients with MDS, classified according to FAB (17 RA, five RARS, 19 RAEB, nine RAEBT, ten CMML) and WHO (ten RA, three RARS, seven RCMD, two RCMD-RS, 11 RAEBI, eight RAEBII, ten CMML, and nine AML) were included in the study. We found increased expression of anti apoptotic bclxL and mcl1 genes and decreased expression of p21 gene in MDS patients. Moreover, we found increased expression of anti-apoptotic mcl1 gene in patients with higher than Intermediate-1 IPSS group. Multivariate analysis confirmed that combined expression of apoptotic caspases 8, 3, 6, 5, 2, 7, and Granzyme B was decreased in MDS patients. Regarding cell cycle regulatory genes expression, we demonstrated increased expression of cyclin D1 in patients with CMML Increased combined expression of cyclins B, C, D1, and D2 was found in patients with cytogenetic abnormalities. The two pathways seem to be interconnected as shown by the positive correlation between CDKs 1, 2, 4, p21 and the level of apoptosis and positive correlation between apoptotic caspase 3 expression and the percentage of S phase cells. In conclusion, our study showed altered expression of genes involved in apoptosis and cell cycle in MDS and increased expression of cyclin D1 in patients with CMML. PMID- 19813014 TI - [Pattern recognition in the differential diagnosis of salivary lymphoepithelial lesions]. AB - The prototype of a salivary lymphoepithelial lesion is the autoimmune disease Sjogren's syndrome with the characteristic lymphoepithelial duct lesions (LEL). The distinction of Sjogren's syndrome from cases with initial transformation into associated marginal zone B-cell lymphoma (MALT type) can be very challenging, whereby the presence of small "halos" can lead to over-diagnosis. The HIV associated cystic lymphoepithelial lesion can be histologically almost identical to Sjogren's syndrome and often needs clinical correlation. The sporadic lymphoepithelial cyst of the parotid gland is a frequent finding and has no clinical consequence; however, this entity needs to be identified and distinguished from the above-mentioned entities. The most frequent diagnosis in resected submandibular glands is chronic-fibrosing sialadenitis, so-called Kuttner's tumour. Altogether, there is a wide spectrum of lymphoepithelial interaction in the area of salivary glands, including biphasic lymphoepithelial tumours with an obligate lymphoid component, epithelial tumours with facultative tumour-associated lymphoid proliferation, and different processes of intraparotid lymph nodes. The immunohistological reaction for pan-keratin can be very helpful for a thorough pattern analysis of the different lymphoepithelial lesions. The relative frequency of the lesions in different salivary glands can also be diagnostically helpful. PMID- 19813015 TI - Functional characterization of Citrus PSY gene in Hongkong kumquat (Fortunella hindsii Swingle). AB - Citrus, rich in carotenoids, is the most important fruit crop based on the total annual production. In the carotenoid biosynthesis pathway, phytoene synthase (PSY, EC 2.5.1.32) catalyzes the dimerization of two molecules of geranylgeranyl pyrophosphate (GGPP) to phytoene and has been shown to be a rate-limiting enzyme for the synthesis of carotenoids. In this study, we investigated catalytic activity of CsPSY from Cara Cara navel orange (Citrus sinensis Osbeck) by heterologous expression in Escherichia coli containing a GGPP-producing plasmid. Moreover, the effects of CsPSY overexpression on carotenoid accumulation were also functionally analyzed in transgenic Hongkong kumquat (Fortunella hindsii Swingle). The resulting transgenic plants produced orange fruits, and extracts from the fruits of four overexpressing plants had a 2.5-fold average increase of phytoene with the content approximately 71.38 microg/g fresh weight. Lycopene, beta-carotene, and beta-cryptoxanthin in transgenic fruits were also markedly increased, whereas the levels of lutein and violaxanthin kept nearly unchanged with 1.1-1.3 folds variation. Transcript levels of carotenoid biosynthetic genes in the CsPSY overexpressed plants remained unaltered except that PDS and ZDS showed a minor increase. This study suggests that CsPSY plays a crucial role in citrus carotenoid biosynthesis and could be used as a means of engineering fruit crop for the production of carotenoids. PMID- 19813016 TI - [Albumin eye drops for treatment of ocular surface diseases]. AB - For more than 10 years eye drops made from autologous serum have become an established therapy to support corneal wound healing in ocular surface diseases, such as persistent epithelial defects and dry eye. Serum eye drops can only be dispensed by institutions which have obtained a specific license from the appropriate local authorities and this applies to only a few institutes in Germany. The main protein component of serum is albumin which is commercially available as a quality controlled medical product from the pharmaceutical industry and could thus be used as an alternative to autologous serum eye drops. The wound healing effect of albumin eye drops has already been demonstrated in vitro. Here we outline the therapeutic principles and present our first clinical experiences with eye drops made from albumin in the treatment of ocular surface disorders. PMID- 19813017 TI - Visual detection of diminutive floral guides in the bumblebee Bombus terrestris and in the honeybee Apis mellifera. AB - Many flowers display colour patterns comprising a large peripheral colour area that serves to attract flower visitors from some distance, and a small central, contrastingly coloured area made up by stamens or floral guides. In this study, we scaled down the size of floral guides to detect the minimal size bumblebees (Bombus terrestris) and honeybees (Apis mellifera) require for guidance. We analyzed the approach and the precise contact of the antennal tips with the floral guide of artificial flowers which precedes landing and inspection. Both bumblebees and honeybees were able to make antennal contact with circular floral guides which were 2 mm in diameter; bumblebees performed better than honeybees and antennated also at floral guides smaller than 2 mm. In discrimination experiments with bumblebees, a minimum floral guide size of 2 mm was required for discrimination between artificial flowers with and without floral guides. With increasing experience bumblebees targeted close to the site of reward instead of making antennal contact with the floral guide, whereas honeybees did not alter their initial behaviour with growing experience. Bumblebees and honeybees spontaneously target diminutive floral guides to achieve physical contact with flowers by means of their antennae which helps them to inspect flowers. PMID- 19813018 TI - The flicker electroretinogram interocular amplitude ratio is a strong prognostic indicator of neovascularization in patients with central retinal vein occlusion. AB - BACKGROUND: To evaluate the prognostic value of interocular amplitude ratio of flicker electroretinogram (ERG) in determining the development of neovascularization in patients with central retinal vein occlusion (CRVO). METHODS: We retrospectively reviewed the data obtained from flicker ERG in 51 CRVO patients. Of these, 22 eyes which had enough follow-up to differentiate ischemic CRVO from nonischemic CRVO were included for data analysis. The flicker ERG was recorded at a 30 Hz frequency after dark adaptation, and ten sweeps were averaged. RESULTS: Eleven eyes were ischemic and 11 eyes were nonischemic. Three amplitude parameters had the potential to explain the type of CRVO. They were amplitude of lesion eye (p = 0.0001), interocular difference of amplitude (p < 0.0001), and interocular ratio of amplitude (p < 0.0001). Both an interocular amplitude difference of -23 microV and interocular amplitude ratio of 60% were very good cutoff points to differentiate ischemic from nonischemic CRVO. Receiver operating characteristic curve analysis revealed that each of the two cutoff values had a sensitivity and specificity of 100%. CONCLUSIONS: Interocular comparison of amplitude is a good solution for avoiding the variability of ERG. An interocular amplitude ratio of flicker ERG of 60% is a succinct, useful parameter in clinical practices for differentiating ischemic from nonischemic CRVO. PMID- 19813019 TI - An EMG frequency-based test for estimating the neuromuscular fatigue threshold during cycle ergometry. AB - The purposes of this investigation were twofold: (1) to determine if the model used for estimating the physical working capacity at the fatigue threshold (PWC(FT)) from electromyographic (EMG) amplitude data could be applied to the frequency domain of the signal to derive a new fatigue threshold for cycle ergometry called the mean power frequency fatigue threshold (MPF(FT)), and (2) to compare the power outputs associated with the PWC(FT), MPF(FT), ventilatory threshold (VT), and respiratory compensation point (RCP). Sixteen men [mean (SD) age = 23.4 (3.2) years] performed incremental cycle ergometer rides to exhaustion with bipolar surface EMG signals recorded from the vastus lateralis. There were significant (p < 0.05) mean differences for PWC(FT) [mean (SD) = 168 (36) W] versus MPF(FT) [208 (37) W] and VT [152 (33) W] versus RCP [205 (84) W], but no mean differences for PWC(FT) versus VT or MPF(FT) versus RCP. The mean difference between PWC(FT) and MPF(FT) may be due to the effects of specific metabolites that independently influence the time and frequency domains of the EMG signal. These findings indicated that the PWC(FT) model could be applied to the frequency domain of the EMG signal to estimate MPF(FT). Furthermore, the current findings suggested that the PWC(FT) may demarcate the moderate from heavy exercise domains, while the MPF(FT) demarcates heavy from severe exercise intensities. PMID- 19813020 TI - Measurement of cytotoxic T lymphocyte activity of human cytomegalovirus seropositive individuals by a highly sensitive coupled luminescent method. AB - A coupled luminescent method (CLM) based on glyceraldehyde-3-phosphate dehydrogenase released from injured target cells was used to evaluate the cytotoxicity of antigen-specific HLA class I-restricted CTLs. In contrast to established methods, CLM does not require the pretreatment of target cells with radioactive or toxic labeling substances. CTLs from healthy HLA-A2 positive donors were stimulated by autologous dendritic cells (DCs) pulsed with HLA-A2 restricted HCMV-pp65 nonamer peptides. HLA-A2 positive T2 cells or autologous monocytes pulsed with HCMV-pp65 nonamer peptide served as target cells. Lysis was detected only in HCMV-pp65-pulsed target cells incubated with CTLs from seropositive donors stimulated by HCMV-pp65-pulsed DCs. After 3 days, stimulation 38% of T2 cells and 17% of monocytes were lysed at an effector to target ratio of 8:1. In conclusion, CLM represents a highly sensitive, fast, material-saving and non-toxic/non-radioactive method for the measurement of antigen-specific CTL cytotoxic activity. PMID- 19813021 TI - Thoracolumbar intervertebral disc calcifications in an 8-year-old boy: case report and review of the literature. AB - An 8-year-old boy presented himself with a nontraumatic acute onset of thoracolumbar pain with an antalgic scoliosis. His past medical history did not reveal any infectious disorder. The back pain kept him from sleep and sports. Imaging revealed calcified intervertebral discs from thoracic intervertebral disc 10-11 to lumbar disc 2-3, with compression of the 11th thoracic root. Conservative treatment with analgesia, nonsteroidal anti-inflammatory drugs, and rest reduced the symptoms within a few days. The boy became asymptomatic within 2 weeks. Magnetic resonance imaging changes diminished in 2 months. Intervertebral disc calcification in children is a rare benign disorder which mostly affects the cervical spine. This is the first case, to our knowledge, of symptomatic intervertebral disc calcification involving the lumbar spine up to lumbar disc 2 3. PMID- 19813022 TI - Significant increase of succinylacetone within the first 12 h of life in hereditary tyrosinemia type 1. AB - INTRODUCTION: In most countries, hereditary tyrosinemia type 1 is not included in routine newborn screening. DISCUSSION: We present the case of a female newborn with prenatal diagnosis of hereditary tyrosinemia type 1 and clear identification of this disorder by succinylacetone measurement in cord blood and peripheral blood immediately after birth. Succinylacetone was 44 micromol/L (norm <5 micromol/L) and increased within 12 h to 87.5 micromol/L. CONCLUSION: With the high toxic potential of downstream metabolites, these data clearly point out the necessity of early nitisinone treatment to prevent symptomatic disease. PMID- 19813023 TI - An intronic polymorphism in the corticotropin-releasing hormone receptor 2 gene increases susceptibility to HBV-related hepatocellular carcinoma in Chinese population. AB - Corticotropin-releasing hormone receptor 2 (CRHR2) plays a role in both the central nervous system (CNS) and the peripheral nervous system. CRHR2 together with its ligands, urocortins (Ucns) and corticotropin-releasing hormone (CRH), functions as a mediator of inflammatory response and inhibitor of angiogenesis. Recently, it has been reported to be expressed in many human cancers. An association between rs2267716 polymorphism in the CRHR2 gene and susceptibility to hepatocellular carcinoma (HCC) was found in patients with chronic hepatitis C virus (HCV) infection. In the present study we analyzed, using a polymerase chain reaction-ligation detection reaction (PCR-LDR), the rs2267716 polymorphism in 364 hepatitis B virus (HBV)-related HCC patients, 196 non-HCC patients with HBV infection, and 404 healthy controls. The aim was to detect the possible association of this single-nucleotide polymorphism (SNP) with susceptibility to HBV-related HCC. Significant differences of rs2267716 allele were detected between HBV-related HCC patients and healthy controls (OR = 1.55, 95% CI 1.13 2.15, P = 0.007) or non-HCC patients with HBV infection (OR = 1.61, 95% CI 1.13 2.31, P = 0.009). These results suggest that the rs2267716 polymorphism in the CRHR2 gene might influence the risk of developing HCC in patients with HBV infection in Chinese population. PMID- 19813025 TI - A comment on the paper: Extended Y chromosome haplotypes resolve multiple and unique lineages of the Jewish Priesthood by M.F. Hammer, D.M. Behar, T.M. Karafet, F.L. Mendez, B. Hallmark, T. Erez, L.A. Zhivotovsky, S. Rosset, K. Skorecki, Hum Genet, published online 8 August 2009. PMID- 19813026 TI - Expression and localization of acrosin inhibitor in boar reproductive tract. AB - Proteinases and proteinase inhibitors play key roles in almost all physiological processes. Proteinase inhibitors are present in all tissues and body fluids. They interfere with the activity of proteinases and thus maintain homeostasis. The main role of proteinase inhibitors in the reproductive tract is the inactivation of prematurely released hydrolytic enzymes from damaged spermatozoa and the protection of reproductive tracts and spermatozoa against proteolytic degradation. In the boar reproductive system, acrosin inhibitors are found in seminal plasma and on spermatozoa. The amino acid sequence of seminal plasma and sperm-associated acrosin inhibitors is 90% identical, and their biochemical properties have been completely resolved. However, their origin and localization have not been fully elucidated. Using rabbit polyclonal antibody, we have studied the expression and localization of the seminal plasma acrosin inhibitor in the boar reproductive tract. The antibody recognizes a 12-kDa band in extracts from the cauda epididymidis, seminal vesicles, prostate, and Cowper's glands, and immunofluorescence has revealed the acrosin inhibitor in the epithelium and lumen of these organs. Gene expression of the acrosin inhibitor has been studied by reverse transcription together with the polymerase chain reaction. Acrosin inhibitor mRNA transcript is detectable in the epididymis, seminal vesicles, prostate, and Cowper's glands. The antibody has localized the acrosin inhibitor on the surface of epididymal and ejaculated spermatozoa in the acrosomal region. In extracts from epididymal and ejaculated spermatozoa, the specific antibody recognizes acrosin inhibitor at 8 kDa and 12 kDa. The presence of acrosin inhibitor on the sperm surface as a protective molecule for receptors mediating the sperm-zona pellucida binding suggests that it is crucial for the reproductive process. PMID- 19813027 TI - Unexpected matrix diseases and novel therapeutic strategies. AB - Within the framework of a broad definition of the extracellular matrix (ECM), this review discusses three genetic disorders in which major pathogenetic features have been traced back to alterations in the levels/activities of matrix components. In each case, disease-associated alterations are found both intra- and extracellularly. The nature of the ECM involvement is surprising, offers an exciting therapeutic opportunity, and deepens our understanding of ECM-cell interactions. The first of these disorders, cherubism, is a case of inflammatory bone loss in the jaws of children for reasons that are surprisingly systemic in nature, considering the local nature of the disease. The primary defect involves an intracellular signaling molecule, but a major pathogenetic component and therapeutic target of the disease is the extracellular cytokine tumor necrosis factor alpha. The second disorder, Knobloch syndrome, is caused by recessive mutations in collagen XVIII. Although this protein has been classified as belonging to a group of structural macromolecules, the consequence of the mutations is impairment of cellular metabolism. The third disorder, infantile hemangioma, is a common tumor of capillary endothelial cells in infancy. The tumor appears within a few days/weeks after birth, grows rapidly over several months, and regresses over several years. The proliferative phase is the result of constitutively high levels of vascular endothelial cell growth factor (VEGF) dependent signaling through VEGF receptor 2 (VEGFR2), but recent studies have led to the surprising conclusion that abnormalities in a cell-surface receptor complex controlling expression of the VEGF decoy receptor VEGFR1 is the underlying cause. PMID- 19813028 TI - Lactoferrin at basal side of mouse mammary epithelium derives in part from stroma cells. AB - Lactoferrin is synthesized by glandular epithelial cells and neutrophils and is also present on both sides of the mammary epithelium. We have studied the origin of lactoferrin detected in the various compartments of mouse mammary tissue. As revealed by immunogold electron microscopy, lactoferrin is present in mammary epithelial cells and in the basal region of the epithelium, associated with connective tissue and stroma cells at all physiological stages studied. A perturbation of protein synthesis or transport after in vitro treatment with cycloheximide or brefeldin A does not abrogate lactoferrin labelling in the basal region of the epithelium. The expression of lactoferrin has also been observed in the fat pads of mammary glands from mice surgically depleted of epithelial cells. The sealing of one teat for 24 h is accompanied by an increase in both the number of stroma cells and the labelling of myoepithelial cells. Thus, the lactoferrin present in the interstitial space of the mouse mammary epithelium originates in part from stroma cells. Possible roles of lactoferrin at the basal side of the mammary epithelium are discussed. PMID- 19813029 TI - Prehospital emergency treatment of palliative care patients with cardiac arrest: a retrolective investigation. AB - BACKGROUND: Today, prehospital emergency medical teams (EMTs) are confronted with emergent situations of cardiac arrest in palliative care patients. However, little is known about the out-of-hospital approach in this situation and the long term survival rate of this specific patient type. The aim of the present investigation was to provide information about the strategic and therapeutic approach employed by EMTs in outpatient palliative care patients in cardiac arrest. METHODS: During a period of 2 years, we retrolectively analysed emergency medical calls with regard to palliative care emergency situations dealing with cardiac arrest. We evaluated the numbers of patients who were resuscitated, the prevalence of an advance directive or other end-of-life protocol, the first responder on cardiac arrest, the return of spontaneous circulation (ROSC) and the survival rate. RESULTS: Eighty-eight palliative care patients in cardiac arrest were analysed. In 19 patients (22%), no resuscitation was started. Paramedics and prehospital emergency physicians began resuscitation in 61 cases (69%) and in 8 cases (9%), respectively. A total of 10 patients (11%) showed a ROSC; none survived after 48 h. Advance directives were available in 43% of cases. The start of resuscitation was independent of the presence of an advance directive or other end-of-life protocol. CONCLUSIONS: Strategic and therapeutic approaches in outpatient palliative care patients with cardiac arrest differ depending on medical qualification. Although many of these patients do not wish to be resuscitated, resuscitation was started independent of the presence of advance directive. To reduce legal insecurity and to avoid resuscitation and a possible lengthening of the dying process, advance directives and/or "Do not attempt resuscitation" orders should be more readily available and should be adhered to more closely. PMID- 19813031 TI - Kinetics and mechanism of exogenous anion exchange in FeFbpA-NTA: significance of periplasmic anion lability and anion binding activity of ferric binding protein A. AB - The bacterial transferrin ferric binding protein A (FbpA) requires an exogenous anion to facilitate iron sequestration, and subsequently to shuttle the metal across the periplasm to the cytoplasmic membrane. In the diverse conditions of the periplasm, numerous anions are known to be present. Prior in vitro experiments have demonstrated the ability of multiple anions to fulfill the synergistic iron-binding requirement, and the identity of the bound anion has been shown to modulate important physicochemical properties of iron-bound FbpA (FeFbpA). Here we address the kinetics and mechanism of anion exchange for the FeFbpA-nitrilotriacetate (NTA) assembly with several biologically relevant anions (citrate, oxalate, phosphate, and pyrophosphate), with nonphysiologic NTA serving as a representative synergistic anion/chelator. The kinetic data are consistent with an anion-exchange process that occurs in multiple steps, dependent on the identity of both the entering anion and the leaving anion. The exchange mechanism may proceed either as a direct substitution or through an intermediate FeFbpA-X* assembly based on anion (X) identity. Our kinetic results further develop an understanding of exogenous anion lability in the periplasm, as well as address the final step of the iron-free FbpA (apo-FbpA)/Fe(3+) sequestration mechanism. Our results highlight the kinetic significance of the FbpA anion binding site, demonstrating a correlation between apo-FbpA/anion affinity and the FeFbpA rate of anion exchange, further supporting the requirement of an exogenous anion to complete tight sequestration of iron by FbpA, and developing a mechanism for anion exchange within FeFbpA that is dependent on the identity of both the entering anion and the leaving anion. PMID- 19813030 TI - Posttranslational regulation of copper transporters. AB - Copper is an essential but potentially harmful trace element involved in many enzymatic processes that require redox chemistry. Cellular copper homeostasis in mammals is predominantly maintained by posttranslational regulation of copper import and export through the copper import proteins hCTR1 and hCTR2 and the copper exporters ATP7A and ATP7B. Regulation of copper uptake and export is achieved by modulation of transporter expression, copper-dependent and copper independent trafficking of the different transporters, posttranslational modifications, and interacting proteins. In this review we systematically discuss the contribution of these different mechanisms to the regulation of copper transport. PMID- 19813032 TI - Hemangioma of the mandible: case report with special emphasis on bone degradation. AB - PURPOSE: Intraosseous hemangioma of the mandible is a rare tumor. In this report, we present a case with a particularly prominent osteolysis and discuss the histological features of bone degradation. MATERIALS AND METHODS: We describe a 64-year-old female patient with a cystic tumor mass of the mandible leading to pathologic bone fracture. X-ray analysis was suggestive for aneurysmatic bone cyst. A segment resection was performed. RESULTS: The histological examination revealed a mesenchymal tumor with numerous capillaries and dilated vessels immunohistochemically being positive for CD31, but not for D2-40. In addition, there was a remarkable increase of osteoclasts that sometimes exhibited Howship's lacunae. The tumor was diagnosed as intraosseous hemangioma with prominent bone degradation. CONCLUSIONS: Based on our histological findings, it should be further investigated whether there could be an association between angiogenesis and osteoclastogenesis as a central pathway leading to bone destruction in the case of intraosseous hemangiomas. PMID- 19813033 TI - Suggestive evidence on chromosomes 2 and 19 for HTR1A-independent linkage of genes to major depression. PMID- 19813034 TI - Extinction memory in the crab Chasmagnathus: recovery protocols and effects of multi-trial extinction training. AB - A decline in the frequency or intensity of a conditioned behavior following the withdrawal of the reinforcement is called experimental extinction. However, the experimental manipulation necessary to trigger memory reconsolidation or extinction is to expose the animal to the conditioned stimulus in the absence of reinforcement. Recovery protocols were used to reveal which of these two processes was developed. By using the crab contextual memory model (a visual danger stimulus associated with the training context), we investigated the dynamics of extinction memory in Chasmagnathus. Here, we reveal the presence of three recovery protocols that restore the original memory: the old memory comes back 4 days after the extinction training, or when a weak training is administered later, or once the VDS is presented in a novel context 24 h after the extinction session. Another objective was to evaluate whether the administration of multi-trial extinction training could trigger an extinction memory in Chasmagnathus. The results evince that the extinction memory appears only when the total re-exposure time is around 90 min independently of the number of trials employed to accumulate it. Thus, it is feasible that the mechanisms described for the case of the extinction memory acquired through a single training trial are valid for multi-trial extinction protocols. Finally, these results are in agreement with those reports obtained with models phylogenetically far apart from the crab. Behind this attempt is the idea that in the domain of studies on memory, some principles of behavior organization and basic mechanisms have universal validity. PMID- 19813035 TI - The intracellular effects of manuka honey on Staphylococcus aureus. AB - The purpose of this study was to investigate the effect of manuka honey on Staphylococcus aureus in order to identify the intracellular target site. The mode of inhibition of manuka honey against S. aureus NCTC 10017 was investigated by determining the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and the effect of time on viability. Structural changes were observed by scanning (SEM) and transmission electron microscopy (TEM) of cells suspended for 4 h at 37 degrees C in 0.05 mM Tris buffer containing 10% (w/v) manuka honey and were compared to cells in buffer alone or buffer containing 10% (w/v) artificial honey (to assess osmotic damage). A bactericidal mode of inhibition for manuka honey on S. aureus was established. Marked structural changes in honey-treated cells were seen only with TEM, where a statistically significant increase in the number of whole cells with completed septa compared to untreated cells were observed (P < 0.05). Structural changes found with TEM suggest that honey-treated cells had failed to progress normally through the cell cycle and accumulated with fully formed septa at the point of cell division without separating. Sugars were not implicated in this effect. The staphylococcal target site of manuka honey involves the cell division machinery. PMID- 19813036 TI - Down-regulation of defense genes and resource allocation into infected roots as factors for compatibility between Fagus sylvatica and Phytophthora citricola. AB - Phytophthora citricola is a wide spread and highly aggressive pathogen of Fagus sylvatica. The hemibiotrophic oomycete infects the roots and establishes a compatible interaction with F. sylvatica. To investigate the transcriptional changes associated with P. citricola infection, 68 custom oligo-microarray measurements were conducted. Hierarchical as well as non-hierarchical clustering was carried out to analyze the expression profiles. Experimental setup includes a time scale covering the biotrophic and necrotrophic stages of interaction as well as comparative analyses of the local and systemic responses. The local reaction of F. sylvatica is characterized by a striking lack of defense gene induction leading to the conclusion that P. citricola escapes the main recognition systems and/or suppresses the host's response. The analysis of the systemic reaction revealed a massive shift in gene expression patterns during the biotrophic phase that is interpreted as evidence of resource allocation into the roots to support the increased sink caused by pathogen growth. Defense genes known to be responsive to salicylic acid (effective against biotrophs), jasmonic acid, and ethylene (effective against necrotrophs and herbivores) are represented on the arrays. All significant changes in gene expression measured for salicylic acid responsive genes were down-regulations in roots and leaves while some jasmonic acid responsive genes showed a very late up-regulation only in leaves, probably caused by the desiccation shortly before plant death. Together, these expression changes could explain the success of the pathogen. PMID- 19813037 TI - Impact of CPAP treatment on cardiac biomarkers and pro-BNP in obstructive sleep apnea syndrome. AB - OBJECTIVE: To evaluate the effect of continuous positive airway pressure (CPAP) therapy on pro-brain natriuretic peptide (BNP) and cardiac markers in patients with obstructive sleep apnea syndrome and normal cardiac function. METHODS: Thirty-three consecutive patients with sleep apnea syndrome were analysed for serum pro-BNP and cardiac markers prior to and after 6 months of CPAP therapy. RESULTS: Twenty five patients had normal (83.3%) while remaining five (16.7%) revealed high pro-BNP values. We did not detect any significant difference between severity of obstructive sleep apnea syndrome and serum pro-BNP levels (p = 0.534). A statistically significant difference was not observed between basal and sixth-month creatine kinase (CK), creatine kinase-MB (CK-MB), troponin I, pro BNP, aspartate transaminase (AST), and CK levels in patients with sleep apnea syndrome (p > 0.05). CONCLUSION: Obstructive sleep apnea syndrome does not induce myocardial damage enough to increase serum pro-BNP, CK, CK-MB, troponin I, and AST levels. Markers sensitive to ischemia could be preferred to evaluate effect of obstructive sleep apnea syndrome. PMID- 19813039 TI - A novel method using sedimentary metals and GIS for measuring anthropogenic change in coastal lake environments. AB - BACKGROUND, AIM AND SCOPE: A new method using sedimentary metals and geographic information system as indicators for assessing temporal and spatial anthropogenic change in estuaries has been applied to a large coastal lake (Lake Macquarie) in New South Wales, Australia. MATERIALS, METHODS AND RESULTS: Two vintages of data (1975 and 2003) on surficial sediment metal (Cd, Cu, Pb and Zn) concentrations combined with (210)Pb core profiles were used to determine past changes in sediment quality and to predict possible future relaxation rates for the entire lake area in response to change in anthropogenic pressure. Sediment cores showed distinct vertical profiles; sedimentation rates in the northern part of the lake were consistent (14 mm year(-1)) over the 55-year period investigated. DISCUSSION AND CONCLUSIONS: Surficial metal concentrations were highest in the 1975 sediment than in the 2003 samples, with the northern part of Lake Macquarie having much greater metal concentrations than the rest of the lake. Past and future declining sedimentary metal concentrations in the northern part of the lake were expected due to the closure of a nearby Pb-Zn smelter; however, possible increases in Cu in the south of the lake to the year 2020 were surprising. The new method presented in this study can assist estuary managers by providing data on past, present and future conditions, which are essential in making informed decisions for the improvement of estuarine systems. PMID- 19813038 TI - Changes of hematocrit and hemoglobin concentration in the cold Himalayan environment in dependence on total body fluid. AB - INTRODUCTION: The organism is exposed to a considerable hypoxic stress at high altitude, and the well-known polyglobulia is an effective strategy to sustain oxygen delivery to the tissue at reduced saturation of hemoglobin. In general, an increasing erythropoiesis is thought to be the reason, although this increase of red blood count can be observed after a short time of altitude exposure and the parameters are expressed as water-depending concentrations. Therefore, the influence of water distribution on hemoglobin (Hb) and hematocrit (Hct) values during a long-term exposure at high altitude was investigated. MATERIALS AND METHODS: Measurements were performed in 12 mountaineers before, during, and either 7/8 or 11/12 days after a Himalaya expedition (26-29 days at 4,850 to 7,600 m altitude). Arriving at 4,850 m an initial increase of Hb and Hct was followed by a short decrease during the first week and a continuous increase during the further stay. RESULTS: In maximum, 131.3% (Hb) and 117.4% (Hct) of the starting point were reached during the fourth week at altitude after the attempt to reach the summit of Broad Peak (8,047 m). Parallel the dehydration in the beginning turned to a hyperhydration at the end of the stay (D(2)O method). DISCUSSION: Erythropoietin rose only temporarily at altitude (max. +11 +1 mU/ml serum). Upon return, Hb and Hct normalized within a few days whereas hemoglobin mass (initially 881+ 44 g, CO-Hb method) was still increased by 13% (p < 0.01). CONCLUSION: In conclusion, a hemoconcentration effect (dehydration) is the reason of the initial peak of Hb and Hct. The further increase can only partially be explained by an absolute increase of Hb and Hct caused by stimulated erythropoiesis. A shift of intravasal fluid to the interstitial space is the other main reason of the observed changes in red blood count. PMID- 19813040 TI - [Psychological therapy as part of an interdisciplinary treatment of chronic back pain]. AB - Providing the patient with a pain model based on the biopsychosocial approach is one of the most important issues in psychological intervention. Illness behaviour is influenced by pain-eliciting and pain-aggravating thoughts. Identification and modification of these thoughts is essential and aims to change cognitive evaluations, emotional processing, and pain-referred behaviour. Improved self monitoring concerning maladaptive thoughts, feelings, and behaviour enables functional coping strategies (e.g. attention diversion and learning to enjoy things) and enhances self-efficacy expectancies. Of special importance is the establishment of an appropriate balance between stress and recreation. Intervention options include teaching relaxation techniques, problem-solving strategies, and specific skills as well as applying appropriate elements of cognitive therapy. The development of alternative cognitive and action-based strategies improves the patient's ability to cope with internal and external stressors. All of the psychological elements are carried out in a group setting. Additionally, individual therapy is offered to treat comorbidities or to support reintegration into the patient's job. PMID- 19813042 TI - Respiratory complications associated with IV zoledronic acid infusion in the treatment of postmenopausal osteoporosis. PMID- 19813043 TI - Relationship between 3-month changes in biochemical markers of bone remodelling and changes in bone mineral density and fracture incidence in patients treated with strontium ranelate for 3 years. AB - SUMMARY: From two randomised controlled trials, it is shown that 3-month changes in biochemical markers of bone formation (bone-specific alkaline phosphatase and C-terminal propeptide of type I procollagen) were associated with 3-year bone mineral density (BMD) changes, but not fracture incidence in patients treated with strontium ranelate. INTRODUCTION: The purpose of this study was to assess if short-term change in biochemical markers of bone remodelling is associated with long-term BMD change and fracture incidence observed during treatment with strontium ranelate. METHODS: From the SOTI and TROPOS trials, bone-specific alkaline phosphatase (BALP), C-terminal propeptide of type I procollagen (PICP), serum C-terminal telopeptides (S-CTX) and urine N-terminal telopeptides of type I collagen (U-NTX) were assessed at baseline and after 3 months. RESULTS: Two thousand three hundred seventy-three women were included in this study. Multiple regression analysis showed that 3-month changes in PICP and BALP but not s-CTX I nor s-NTX I were significantly (p < 0.001) associated with 3-year BMD changes at the lumbar spine and the femoral neck. Changes in s-CTX I, PICP and BALP were significantly associated with change in total proximal femur BMD. Changes in biochemical markers explain less than 8% of the BMD changes. The 3-month changes in BALP, PICP s-CTX I and s-NTX I were not significantly associated with fracture incidence. CONCLUSIONS: Short-term changes in biochemical markers of bone formation are associated with future BMD changes in patients treated with strontium ranelate, suggesting a bone-forming activity of this treatment, but are not appropriate to monitor the efficacy of strontium ranelate at the individual level. PMID- 19813044 TI - Suppressive effects of 1,4-dihydroxy-2-naphthoic acid administration on bone resorption. AB - SUMMARY: The main component of the metabolic by-products of fermentation by Propionibacterium freudenreichii ET-3 is 1,4-dihydroxy-2-naphthoic acid (DHNA), which has a naphthoquinone skeleton, as in vitamin K2. This study showed that DHNA improved bone mass reduction with osteoporosis model mice caused by FK506. INTRODUCTION: Growth of the intestinal bacterium Lactobacillus bifidus is specifically facilitated by DHNA. The present study used osteoporosis model mice to investigate the effects of DHNA on bone remodeling. METHODS: FK506, an immunosuppressant, was used to prepare osteoporosis model mice. Thirty mice were divided into three groups: FK group, FK+DHNA group, and control group. In the FK group, FK506 was administered to induce bone mass reduction. In the FK-DHNA group, FK506 and DHNA were administered concurrently to observe improvements in bone mass reduction. To ascertain systemic and local effects of DHNA, we investigated systemic pathological changes in colon, kidney function and cytokine dynamics, and morphological and organic changes in bone and osteoclast dynamics as assessed by culture experiments. RESULTS: Compared to the FK group without DHNA, colon damage and kidney dysfunction were milder for FK+DHNA group, and production of inflammatory cytokines (interleukin (IL)-1beta, IL-6 and tumor necrosis factor (TNF)-alpha) was more suppressed. Furthermore, compared to the group without DHNA, histological analyses and radiography showed that bone resorption was suppressed for the DHNA group. Culture experiments using osteoclasts from murine bone marrow showed osteoclast suppression for the DHNA group compared to the group without DHNA. CONCLUSION: These results show that DHNA has some effects for improving bone mass reduction caused by FK506. PMID- 19813045 TI - Low-energy distal radius fractures in middle-aged and elderly men and women--the burden of osteoporosis and fracture risk : A study of 1794 consecutive patients. AB - One third of 218 men and half of 1,576 women with low-energy distal radius fractures met the bone mineral density (BMD) criteria for osteoporosis treatment. A large proportion of patients with increased fracture risk did not have osteoporosis. Thus, all distal radius fracture patients >or=50 years should be referred to bone densitometry. INTRODUCTION: Main objectives were to determine the prevalence of patients with a low-energy distal radius fracture in need of osteoporosis treatment according to existing guidelines using T-score or=50 years were included. BMD was assessed by dual energy X-ray absorptiometry (DXA) at femoral neck, total hip, and lumbar spine (L2-L4). The WHO fracture risk assessment tool (FRAX(R)) was applied to calculate the 10-year fracture risk. RESULTS: T-scores or=50 years should be referred to bone densitometry, and if indicated, offered medical treatment. PMID- 19813046 TI - Comparison of the Bath Ankylosing Spondylitis Radiology Index and the modified Stoke Ankylosing Spondylitis Spine Score in Turkish patients with ankylosing spondylitis. AB - The objective of the present study was to compare two radiographic scoring methods (the modified Stoke Ankylosing Spondylitis Spine Score (mSASSS) and the Bath Ankylosing Spondylitis Radiology Index-spine (BASRI-spine)) in terms of reliability, construct validity, and feasibility in Turkish ankylosing spondylitis (AS) patients. The study involved seventy-four patients. The patients were evaluated with 100-mm visual analog scale (VAS) for pain, global assessment of patient, and global assessment of doctor. The laboratory evaluations of patients comprised erythrocyte sedimentation rate and serum C-reactive protein. Bath AS Disease Activity Index (BASDAI), Bath AS Functional Index (BASFI), Bath AS Metrology Index (BASMI), and Bath AS Radiology Index (BASRI) were calculated. Bilateral cervical, lumbar spine, and anteroposterior pelvis radiographs of all patients were obtained and evaluated by two radiologists. Each radiograph was scored by two scoring methods, mSASSS and BASRI-spine, and these methods were tested according to the aspects of the Outcome Measures in Rheumatology Clinical Trials filter: reliability, construct validity, and feasibility. The BASRI-spine reached intra- and interobserver intraclass correlation coefficient (ICC) of 0.726 and 0.689, respectively. The mSASSS scores more reliable, with ICC of 0.831 and 0.840, respectively. The BASMI and BASFI correlated significantly with the two scoring systems, respectively (mSASSS r: 0.557, r: 0.319; BASRI-spine r: 0.605, r: 0.285). For the two methods, the magnitude of the correlation with disease duration was similar (mSASSS p < 0.01 and BASRI p < 0.01), but no significant correlation was observed when compared to the BASDAI. It is known that the BASRI-spine is a feasible method that reliably detects damage in patients with AS. However, the present authors believe that, in AS patients, mSASSS should be the radiological scoring method to choose because of less radiation exposure, along with excellent intra- and interobserver reliability. PMID- 19813048 TI - Is the Irish bedside best? AB - BACKGROUND: The use of the bedside to teach the art of clinical medicine is controversial. Rising student numbers can limit patient availability. Studies examining inpatient attitudes to bedside teaching are few. AIMS: We examined inpatients' attitudes to bedside teaching of undergraduate medical students. METHODS: The study was carried out in a 439-bed teaching hospital. A questionnaire, numerically scored (0-10), was prospectively administered to 102 consecutive patients involved in bedside teaching of undergraduate medical students. RESULTS: The results were available from 92 patients. Patients enjoyed the teaching process (mean score 9.13 +/- 1.16) and benefited from a better understanding of their illness (7.11 +/- 2.57). Patients appreciated their role in educating future doctors (mean score 9.52 +/- 1.11) but demonstrated less confidence in their personal contribution to the teaching process (7.81 +/- 1.89). CONCLUSIONS: Inpatients are very willing participants in bedside teaching of undergraduate medical students. PMID- 19813047 TI - Methodological quality of systematic reviews analyzing the use of laser therapy in restorative dentistry. AB - The purpose of this study was to identify systematic reviews (SRs) that compared laser with other dental restorative procedures and to evaluate their methodological quality. A search strategy was developed and implemented for MEDLINE, the Cochrane Library, LILACS, and the Brazilian Dentistry Bibliography (1966- 2007). Inclusion criteria were: the article had to be an SR (+/- meta analysis); primary focus was the use of laser in restorative dentistry; published in English, Spanish, Portuguese, Italian, German. Two investigators independently selected and evaluated the SRs. The overview quality assessment questionnaire (OQAQ) was used to evaluate methodological quality, and the results were averaged. There were 145 references identified, of which seven were SRs that met the inclusion criteria (kappa=0.81). Of the SRs, 71.4% appraised lasers in dental caries diagnosis. The mean overall OQAQ score was 4.4 [95% confidence interval (CI) 2.4- 6.5]. Of the SRs, 57.1% had major flaws, scoring < or = 4. SR methodological quality is low; therefore, clinicians should critically appraise them prior to considering their recommendations to guide patient care. PMID- 19813049 TI - Second toe swelling: Nora's lesion or glomus tumour, case report and literature review. AB - INTRODUCTION: We report a rare case of bizarre parosteal osteochondromatous proliferation (BPOP, Nora's lesion) of the right second toe in a 60-year-old man who presented with painful, bluish and bulbous swelling of the right second toe without any break in the skin. INVESTIGATIONS: Physical examination, plain radiography and magnetic resonance imaging (MRI) scan of the foot, histology of the excised tissue. DIAGNOSIS: Bizarre parosteal osteochondromatous proliferation. MANAGEMENT: Radiographs showed a calcified/ossified lesion adjacent to the tuft of the terminal phalanx of the second toe. MRI showed a small low signal nodule on T1- and T2-weighted images in a subungual position adjacent to the terminal phalanx with sclerosis. The second toe was excised and the histology from excised tissue was consistent with "bizarre parosteal osteochondromatous proliferation". There was no malignant change on histology. Local excision of the entire lesion was done and there has been no recurrence to date. CONCLUSION: BPOP, although a benign lesion, behaves aggressively with rapid growth and has a high risk of local recurrence after local resection (Nora et al. in Am J Surg Pathol 7(3):245-250, 1983; Meneses et al. in Am J Surg Pathol 17(7):691-697, 1993). Its clinical presentation can be confused with glomus tumour, subungual exostosis and enchondroma. The distinguishing features of BPOP and several relevant different differential diagnoses are discussed in this case report. PMID- 19813050 TI - The cervical spine of professional front-row rugby players: correlation between degenerative changes and symptoms. AB - BACKGROUND: Injuries to the cervical spine (C-spine) are among the most serious in rugby and are well documented. Front-row players are particularly at risk due to repetitive high-intensity collisions in the scrum. AIM: This study evaluates degenerative changes of the C-spine and associated symptomatology in front-row rugby players. MATERIALS AND METHODS: C-spine radiographs from 14 professional rugby players and controls were compared. Players averaged 23 years of playing competitive rugby. Two consultant radiologists performed a blind review of radiographs evaluating degeneration of disc spaces and apophyseal joints. Clinical status was assessed using a modified AAOS/NASS/COSS cervical spine outcomes questionnaire. RESULTS: Front-row rugby players exhibited significant radiographic evidence of C-spine degenerative changes compared to the non-rugby playing controls (P < 0.005). Despite these findings the rugby players did not exhibit increased symptoms. CONCLUSION: This highlights the radiologic degenerative changes of the C-spine of front-row rugby players. However, these changes do not manifest themselves clinically or affect activities of daily living. PMID- 19813051 TI - First Irish delivery following sequential, two-stage embryo and blastocyst transfer. AB - BACKGROUND: The timing of embryo transfer (ET) after in vitro fertilisation (IVF) remains controversial, and there are no reliable guidelines available to prospectively identify which patients would benefit from either day-3 or blastocyst transfer. While blastocyst transfer is generally favoured over day-3 transfers, very few IVF patients get both in the same treatment cycle. CASE DESCRIPTION: We report on a 35.5-year-old female with tubal factor infertility who underwent IVF, which included transfer of a fresh day-3 embryo and a thawed blastocyst frozen at day 6. Transfer occurred on two separate days (days 3 and 6) in a two-stage/dual catheter fashion and resulted in a healthy term singleton livebirth. CONCLUSIONS: While combined day-3 and day-5 ET has been available elsewhere for several years, this is the first description of its successful application in Ireland and confirms the effectiveness of coordinated two-stage transfer in a single IVF treatment cycle. PMID- 19813052 TI - Response to the article entitled "Endovascular repair of a traumatic axillary artery pseudoaneurysm," by R. M. Kumar et al. PMID- 19813054 TI - Antibacterial activity of phenolic compounds against the phytopathogen Xylella fastidiosa. AB - Xylella fastidiosa is a pathogenic bacterium that causes diseases in many crop species, which leads to considerable economic loss. Phenolic compounds (a group of secondary metabolites) are widely distributed in plants and have shown to possess antimicrobial properties. The anti-Xylella activity of 12 phenolic compounds, representing phenolic acid, coumarin, stilbene and flavonoid, was evaluated using an in vitro agar dilution assay. Overall, these phenolic compounds were effective in inhibiting X. fastidiosa growth, as indicated by low minimum inhibitory concentrations (MICs). In addition, phenolic compounds with different structural features exhibited different anti-Xylella capacities. Particularly, catechol, caffeic acid and resveratrol showed strong anti-Xylella activities. Differential response to phenolic compounds was observed among X. fastidiosa strains isolated from grape and almond. Elucidation of secondary metabolite-based host resistance to X. fastidiosa will have broad implication in combating X. fastidiosa-caused plant diseases. It will facilitate future production of plants with improved disease resistance properties through genetic engineering or traditional breeding approaches and will significantly improve crop yield. PMID- 19813055 TI - Effect of smoking on cough reflex sensitivity in humans. AB - Despite the importance of cigarette smoking as a cause of respiratory symptoms and disease, until recently there has been little attention on the effect of smoking on cough reflex sensitivity. Recent studies have shown that in otherwise healthy smokers, cough reflex sensitivity is diminished relative to that of nonsmokers. One proposed mechanism, chronic cigarette smoke-induced desensitization of airway cough receptors, is supported by the demonstration that smoking cessation leads to prompt enhancement of cough reflex sensitivity, even after many years of smoking. Furthermore, resumption of cigarette smoking promptly results in suppression of cough reflex sensitivity. These observations have introduced the concept that cough reflex sensitivity in humans is a dynamic phenomenon, able to be modulated by the presence or absence of stimuli such as cigarette smoke, even after prolonged exposure. PMID- 19813056 TI - Microarray-based identification of gonad transcripts differentially expressed between lines of Pacific oyster selected to be resistant or susceptible to summer mortality. AB - Summer mortality of the Pacific oyster Crassostrea gigas is the result of a complex interaction between oysters, their environment, and pathogens. Heredity appears to be a major factor determining the sensitivity of oysters to summer mortality, allowing resistant (R) and susceptible (S) lines to be produced. We conducted genome-wide expression profiling of R and S gonads during the 3-month period preceding a summer mortality event, using a cDNA microarray that we designed. ANOVA analysis revealed that 34 genes were differentially expressed between R and S lines on four dates preceding the mortality event. Annotation of some of these genes highlights reproduction and its allocation and antioxidant defenses as the main pathways that operate differentially between R and S lines. This transcriptional analysis provides new indications to define markers for quantitative trait loci searches and functional studies and evaluate the potential role of each gene in the resistance to summer mortality. PMID- 19813057 TI - Silicatein genes in spicule-forming and nonspicule-forming Pacific demosponges. AB - Silicatein genes are known to be involved in siliceous spicule formation in marine sponges. Proteins encoded by these genes, silicateins, were recently proposed for nanobiotechnological applications. We studied silicatein genes of marine sponges Latrunculia oparinae collected in the west Pacific region, shelf of Kuril Islands. Five silicatein genes, LoSilA1, LoSilA1a, LoSilA2, and LoSilA3 (silicatein-alpha group), LoSilB (silicatein-beta group), and one cathepsin gene, LoCath, were isolated from the sponge L. oparinae for the first time. The deduced amino acid sequence of L. oparinae silicateins showed high-sequence identity with silicateins described previously. LoCath contains the catalytic triad of amino acid residues Cys-His-Asn characteristic for cathepsins as well as motifs typical for silicateins. A phylogenetic analysis places LoCath between sponge silicateins beta and L-cathepsins suggesting that the LoCath gene represents an intermediate form between silicatein and cathepsin genes. Additionally, we identified, for the first time, silicatein genes (AcSilA and AcSilB) in nonspicule-forming marine sponge, Acsmall a, Cyrillicnthodendrilla sp. The results suggest that silicateins could participate also in the function(s) unrelated to spiculogenesis. PMID- 19813058 TI - Chemical signals in the stingless bee, Frieseomelitta varia, indicate caste, gender, age, and reproductive status. AB - Chemical compounds on the cuticle are a rich source of information used during interactions among social insects. Despite the multitude of studies on these substances and their function in ants, wasps, and honeybees, little is known about this subject in stingless bees (Hymenoptera: Apidae, Meliponini). We studied the chemical composition of the cuticle of the stingless bee, Frieseomelitta varia, by gas chromatography-mass spectrometry (GC-MS), to investigate potential chemical variation among castes, gender, age, and reproductive status. We found differences in the cuticular hydrocarbon composition among workers, males, and queens, recording both qualitative and quantitative differences among individuals of different ages and gender. The cuticle of physogastric queens presented a chemical profile that was distinct from all other groups in the analysis, with high relative abundances of alkenes and alkadienes with 27, 29, and 31 carbon atoms. We discuss the possibility that these compounds signal a queen's presence to the colony, thereby initiating all vital worker-queen interactions. PMID- 19813059 TI - Gastric cancer prognosis: strong correlation between incidence and survival. PMID- 19813061 TI - KRAS mutation and microsatellite instability: two genetic markers of early tumor development that influence the prognosis of colorectal cancer. AB - INTRODUCTION: We examined two genetic markers established early in colorectal tumor development, microsatellite instability (MSI) and mutation of the KRAS proto-oncogene, to see if these genetic changes influence metastatic disease progression and survival. PATIENTS AND METHODS: MSI and KRAS mutation status were assessed in 532 primary adenocarcinomas (stage I-IV) from patients treated by colon resection. Median follow-up was 4.1 years (range 0-13.3 years) overall, 5.4 years for survivors. RESULTS: MSI and KRAS mutation were detected in 12 and 36% of cases, respectively. MSI was more common in early-stage disease (I, 15%; II, 21%; III, 10%; IV, 2%; P = 0.0001). Prevalence of KRAS mutation did not vary with stage (I, 36%; II, 34%; III, 35%; IV, 40%; P = ns). Disease-specific survival was far superior for MSI tumors than for microsatellite stability (MSS) tumors (5 year survival 92 vs. 59%, P < 0.0001). KRAS mutation was a marker of poor survival (5-year survival 55 vs. 68%, P = 0.0002). Using Cox regression analysis MSI, KRAS mutation, and stage were strong independent predictors of survival in the entire patient population. A high-mortality group with MSS/KRAS-mutant tumors was identified within the stage I and II cohort. CONCLUSIONS: MSI and KRAS mutation provide fundamental genetic signatures influencing tumor behavior across patient subsets and stages of tumor development. PMID- 19813062 TI - Late-onset radial nerve palsy associated with conservatively managed humeral fracture. A case report and suggested classification system. AB - Radial nerve palsy can occur with humerus fracture, either at the time of injury (primary) or during reduction (secondary). Late-onset radial nerve palsy (not immediately related to injury or reduction) has been very seldom reported in the English literature. We describe a case of late-onset radial nerve palsy, which developed 9 weeks after an attempted closed management of a midshaft humerus fracture. Exploration of the nerve was performed. The radial nerve was found to be stretched over the ends of the fracture. Open reduction and external fixation of the fracture with mobilization of the nerve from the fracture site lead to complete return of radial nerve function occurring by 3 months. We recommend exploration of cases of late-onset radial nerve palsy in contrast to primary or secondary radial nerve palsy, which can be treated conservatively. Our experience suggests that the cause of the palsy is a continuous ongoing pathology and not a single time event as in primary or secondary cases. Radial nerve palsies associated with humeral fracture should be classified as either primary (at the time of injury), secondary (at the time of reduction), or late onset (not related to either injury or reduction). PMID- 19813063 TI - How do physicians conduct medication reviews? AB - BACKGROUND: Medication reviews are recommended annually for older patients. A medication review is a discussion of a patient's complete set of medications, but the actual content of a review is not well specified. The medical literature suggests that it is an exhaustive evaluation, but what physicians actually ask about their patients' medication regimens has been little studied. OBJECTIVE: To describe what physicians do when they review medications in the office setting. METHODS: Qualitative content analysis of audio-taped encounters between 100 patients aged 65 and older and 28 primary care physicians in two health care systems in Sacramento, California. RESULTS: Physicians use a combination of non mutually exclusive strategies when reviewing chronic medications that include: (1) efforts to obtain a complete list of patient medications (36% of visits), (2) discussion of a topic related to the management of each of a patient's chronic medications (47% of visits), and (3) sequential discussion of the majority of a patient's medications without intervening discussion (45% of visits). Of 10 medication management topics that were discussed in medication reviews, a mean of 1.5 topics (SD = 1.7, range 0-7) were mentioned for each medication, with efficacy and directions being most common. Physicians conducted a sequential discussion that included discussion of each of a patient's medications in only 32% of visits. CONCLUSIONS: Comprehensive discussions about chronic medications are uncommon in routine practice. Practical conceptualization of what constitutes a physician-conducted medication review is needed. PMID- 19813064 TI - Succinylcholine and morbid obesity: the debate continues... PMID- 19813067 TI - Message from the past president of the American Hernia Society. PMID- 19813066 TI - Anti-ERBB2 sh-RNA suppress both cell growth and tumor growth in ERBB2 overexpressing upper gastrointestinal adenocarcinomas. AB - INTRODUCTION: ERBB2 is overexpressed in 15-25% of upper gastrointestinal adenocarcinomas. We use a stable lentiviral shRNA model to demonstrate that ERBB2 suppression in upper gastrointestinal adenocarcinomas with documented ERBB2 amplification effectively decreases ERBB2 protein levels and decreases cell viability. Further, we evaluate tumor growth of cells treated with the ERBB2 shRNA. METHODS: Three upper gastrointestinal adenocarcinoma cells lines with varying ERBB2 levels were treated with one of three separate lentiviral green fluorescent protein (GFP)-labeled ERBB2 shRNA vectors or a nonsilencing control shRNA vector for 6 h. Protein levels on day 6 and cell viability was evaluated on days 3-10. A xenograft in vivo experiment was performed using OE19 cells pretransduced with ERBB2 shRNA to evaluate tumor growth. RESULTS: ERBB2 protein levels decreased by 80%. ERBB2 knockdown significantly decreased cell viability in cell lines with high ERBB2 levels. In vivo tumor growth was suppressed in ERBB2-shRNA-treated groups. CONCLUSION: ERBB2 suppression based on a stable lentiviral shRNA transfection system effectively decreases cell viability in cell lines with amplification of ERBB2 as compared to cell lines without overexpression. ERBB2 knockdown significantly decreases tumor growth in vivo. ERBB2-directed therapy may be of benefit in the subset of patients with gastrointestinal adenocarcinomas exhibiting overamplification of ERBB2. PMID- 19813068 TI - The heart in sporadic inclusion body myositis: a study in 51 patients. AB - The purpose of this study was to explore the prevalence and nature of cardiac abnormalities in sporadic inclusion body myositis (sIBM). Fifty-one sIBM patients were cross-sectionally studied using history-taking, physical examination, measurements of serum creatine kinase activity, the MB fraction (CK-MB), cardiac troponin T (cTnT) and I (cTnI), a 12-lead electrocardiogram (ECG) and 2 dimensional echocardiography. Present cardiac history was abnormal in 12 (24%) out of 51 patients, 12 (24%) patients had abnormalities on ECG, mostly aspecific, and in 12 (24%) patients the echocardiograph showed abnormalities. Elevated CK-MB was present in 42 (82%) patients and 40 (78%) had an elevated cTnT in the absence of acute cardiac pathology. In contrast, in one patient (2%) cTnI was elevated. There was no apparent association between elevated biomarkers, ECG or echocardiographic abnormalities. The prevalence of cardiac abnormalities in sIBM does not seem to be higher than would be expected in these elderly patients. Elevated CK-MB and cTnT levels are common, in contrast to cTnI, but do not reflect cardiac pathology. PMID- 19813069 TI - The number and nature of emergency department encounters in patients with deep brain stimulators. AB - Deep brain stimulation (DBS) has become an increasingly common modality for control of several neurological disorders such as Parkinson's disease, dystonia, essential tremor (ET), and others. Our experience has demonstrated the need for emergency physicians to familiarize themselves with the potential complications of the DBS device as well as the device itself. Therefore, our aim in this paper was to elucidate the number and nature of DBS and non-DBS presentations to the emergency department (ED) and to educate and familiarize ED physicians about DBS devices and their potential complications. We also aimed to devise a simple protocol for DBS management so that all ED physicians would have access to the knowledge or referral capabilities when managing a DBS patient. The objective of the present study was to review the number and nature of ED encounters in patients with deep brain stimulation (DBS) devices implanted for movement and neuropsychiatric disorders. METHODS: The series of encounters reviewed included 215 unique patients with DBS implantation who were identified using an IRB approved database and a paper chart review. Patients in the study included those implanted at University of Florida (UF), as well as those implanted at outside institutions, so long as they were followed at UF. The cohort included n = 215 DBS patients. 25.6% of all 215 patients presented to the ED at least once, with the most common presentation occurring as a result of a decline in mental status when taking into account all visits (6%). Reasons for presentation to the ED included neurological (54.6%), infections/hardware issues (27.9%), orthopedic/focal problems (10.5%), and medical issues (7%). In total, 29 patients arrived at the ED for DBS related issues (23.2%). Of those who presented to the ED (n = 55), the average age was 53.1 (range 10-80 years). Headache was the most common complaint within the neurological category (22.1%), followed by change in mental status (15.1%), and syncope (9.3%). When examining the data by ED diagnosis, change in mental status occurred most commonly in Parkinson's disease (19.6%). Falls were most common in essential tremor (27.2%), and headache occurred most commonly in the dystonia group (52.1%). Across all diseases, mental status change was the most common indication for an ED encounter (6%). Parkinson disease patients most commonly presented with altered mental status (8%), essential tremor patients revealed a high preponderance of falls (6.5%), and dystonia patients tended to present with headache (7.1%). It was concluded that a large number of patients with DBS will present to the ED for many reasons, the majority of which will not be direct complications of their DBS device. Neurological issues were the most common chief complaint, with individual differences depending on the underlying disease. It is important for ED physicians to consider non-DBS related complaints in the presentation of these unique patients since these issues comprise the majority of the ED visits. However, when properly evaluating these patients, management of their DBS device, or referrals to neurosurgery and neurology, if necessary, are imperative. In addition to device management, regular ED standards of care should apply to this special cohort of patients. PMID- 19813070 TI - Asperger's syndrome and high-functioning autism: language, motor and cognitive profiles. AB - The objective of this study is to compare the cognitive profile, the motor and language functioning and the psychosocial adaptation of children with Asperger syndrome (AS) and with high-functioning autism (HFA). Subjects were recruited through the department Autism and Developmental Disorders of the Heckscher Klinikum. To be included in the study, the full-scale-IQ had to be at least 80. Subjects with AS had to have a normal early language development and subjects with HFA a clear delay in language development, as reported by their parents. The sample consisted of 57 children with Asperger syndrome and 55 children with high functioning autism. The mean age of the children was 10 years. All subjects were examined with a standardised test battery. Children with AS had a higher full scale-IQ than children with HFA. This was due to a higher verbal-IQ. There were no significant differences in the performance-IQ. At a mean age of 10 years, subjects with AS had better language skills than subjects with HFA, but at least 30% showed clear receptive language problems. Motor problems were present in about 50% of the children with AS and HFA. The level of psychosocial adaptation was clearly reduced, but was comparable for the two groups. The differences in verbal-IQ and language skills between the two groups could be explained through the definition of the syndromes. The presence of language problems in the subjects with AS at age 10, the comparable degree of motor impairment and level of psychosocial adaptation question the validity of the distinction between AS and HFA within the category of pervasive developmental disorders. PMID- 19813071 TI - Coalitions and male-male behavior in Alouatta palliata. AB - Coalitions influence the establishment and maintenance of social relationships among males in primate species. In this study, we compare the social behavior of males between two groups of Alouatta palliata: a group that was recently taken over by a coalition of two males (Mt), and a group that had a stable composition for at least 9 months (Rh). We predicted that coalition partners would be more cooperative and less competitive than dyads formed by immigrant and long-term resident males, and dyads formed by long-term resident males. Additionally, we predicted that these dyadic trends should be reflected in more competition and less cooperation in the group that was taken over. As predicted, the coalition partners of Mt showed the highest levels of cooperation among all dyads and the second lowest rate of agonism. Cooperation was higher in the group that had a stable composition. Results from this study suggest that the social relationships of male mantled howlers vary as a function of familiarity between males and that in the context of coalitionary takeovers, coalitionary males are highly cooperative. Cooperation is lower in groups recently taken over and competition is more intense, perhaps as a consequence of the process of establishment and reorganization of power relationships within some dyads. In the future, we must determine the frequency of coalitionary takeovers in this population and assess its ultimate consequences for male-male social relationships. PMID- 19813072 TI - 18F-FDG-PET findings of rare case of nonsecretory plasmablastic myeloma. AB - A 61-year-old woman presented with pancytopenia and underwent a bone marrow biopsy. The patient was diagnosed with nonsecretory myeloma (plasmablastic type) based on both the bone marrow biopsy findings and her laboratory data. Fluorine 18 fluorodeoxyglucose-positron emission tomography ((18)F-FDG-PET) was performed prior to chemotherapy showing diffuse bone marrow uptake, splenic uptake, and focal uptake of the right anterior chest wall. The patient underwent an (18)F-FDG PET examination to evaluate the curative effects after three cycles of chemotherapy, and no abnormal uptake on (18)F-FDG-PET was found. Bone marrow biopsy to evaluate the curative effect showed no viable tumor cells. We present a rare case of nonsecretory plasmablastic myeloma detected by (18)F-FDG-PET. PMID- 19813073 TI - Low incidence of methylation of the promoter region of the FANCF gene in Japanese primary breast cancer. AB - PURPOSE: The link between BRCA1 dysfunction and basal-like breast cancer or triple-negative breast cancer (TNBC) has been suggested; however, the associations of other factors involved in the Fanconi anemia (FA)/BRCA pathway with the pathogenesis of basal-like breast cancer remain unidentified. FANCF protein is a component of the FA core complex. The methylation of CpG islands in the FANCF gene plays an important role in occurrence of ovarian cancer and also is an important regulator of cisplatin sensitivity of ovarian cancer. The purpose of this study is to investigate the frequency of FANCF methylation, and to discuss its involvement in the pathogenesis of TNBC and its potency as a predictor of cisplatin sensitivity for breast cancer. METHODS: The methylation of the FANCF gene promoter was investigated, using methylation-specific PCR, in genomic DNA of 99 invasive breast carcinoma specimens obtained from Japanese patients. RESULTS: FANCF methylation was recognized in only 4 of 99 cases (4.0%). No significant correlation was found between FANCF methylation and the expression of ER, PR, HER2, and TNBC. CONCLUSIONS: FANCF methylation is a rare event in Japanese primary invasive breast cancer. This suggests it is not involved in the pathogenesis of TNBC, and it could not be used as a predictor of cisplatin sensitivity in breast cancer. PMID- 19813074 TI - Synthesis, spectroscopic studies and biological activity of a novel nucleopeptide with Moloney murine leukemia virus reverse transcriptase inhibitory activity. AB - In this work, we report the synthesis of an alternate nucleo-alpha,epsilon peptide based on L: -lysine moieties, an in vitro study of its biological activity, and spectroscopical binding studies between the novel nucleopeptide and Moloney murine leukemia virus reverse transcriptase as well as RNA. An alternate homothymine hexamer was synthesized by a straightforward solid phase route starting from commercial materials, purified by RP-HPLC and characterized by ESI MS. The efficiency of the novel nucleo-alpha,epsilon-peptide in interfering with the reverse transcription of eukaryotic mRNA and the noteworthy enzymatic resistance demonstrated by specific assays are in favor of the employment of this nucleopeptide in novel biomedical strategies. PMID- 19813075 TI - Concomitant rectal cancer and abdominal aortic aneurysm: a management strategy. AB - The simultaneous occurrence of abdominal aortic aneurysm and rectal cancer is uncommon but represents a therapeutic dilemma. We report two patients in whom endovascular stenting of the aneurysm was not feasible. These patients were managed by an initial retroperitoneal aortic repair followed a few weeks later by an ultra low anterior resection. PMID- 19813076 TI - Concomitant abdominal aortic aneurysm and rectal cancer: a treatment dilemma. PMID- 19813077 TI - Modified Hanley procedure for management of complex horseshoe fistulae. AB - PURPOSE: Horseshoe fistulae are challenging due to configuration and sphincter involvement. In view of the new treatment options for fistulae (e.g. collagen plug), aim of our study was to review the outcome of patients primarily treated with a traditional approach. METHODS: We retrospectively reviewed patients who presented between 2003 and 2008, with a posterior horseshoe abscess/fistula and were treated with a modified Hanley procedure and seton management. Excluded were Crohn's disease, fistulae from malignancy/surgical complications, and other treatment methods. Data collection included demographics, duration of the disease, and of the treatment, outcome, and incontinence. RESULTS: Twenty-three patients (M/F 20/3) were analyzed. Mean age was 50.3 + or - 10.2 years, median symptom duration 24 months; three patients (19%) had previously received colostomies without resolution. All patients received a posterior midline cutting seton. The average total number of setons was 3 + or - 1.3 (range 2-7), with removal after 1.6 + or - 1.2 months upon resolution of induration and suppuration. The cutting seton was tightened in monthly intervals on average 4.9 + or - 2.2 times. Follow-up was 15.4 + or - 9.7 months: 91.3% patients had complete healing time within 8.1 + or - 4.5 months; 19/23 patients recovered fast, and were able to work no later than 4 weeks postoperatively (2 patients retired, 2 on disability for other reasons). No incontinence was found. CONCLUSIONS: Fecal diversion alone did not resolve horseshoe fistulae. A modified Hanley procedure with drainage of the deep postanal space and cutting and draining setons proved to be safe, successful, and did not result in complaints of fecal incontinence. Completion of the treatment took months, but patients remained functional even with setons in place. PMID- 19813078 TI - Utility of functional diffusion maps to monitor a patient diagnosed with gliomatosis cerebri. AB - Diffusion-weighted magnetic resonance imaging (DWI) is a sensitive imaging biomarker for tumor cellularity. Functional diffusion maps (fDMs), which examine voxel-by-voxel changes in the apparent diffusion coefficient (ADC) calculated from serial DWIs, have previously been applied to regions of contrast enhancement; however, application of fDMs to non-enhancing brain tumors has not been pursued. In this case study we demonstrate the utility of applying fDMs to regions of abnormal FLAIR signal intensity in a patient diagnosed with gliomatosis cerebri: a relatively rare, infiltrative, non-enhancing brain tumor. The absolute volume of hypercellularity extracted from fDMs was useful in tracking tumor growth, which correlated in time with a progressive decline in neurological status despite no change in traditional magnetic resonance images. Results of this study demonstrate the value of fDMs, applied to regions of FLAIR abnormal signal intensity, for localizing regions of hypercellularity and for monitoring overall tumor status. PMID- 19813079 TI - Evaluation of inhibitory activities of plant extracts on production of LPS stimulated pro-inflammatory mediators in J774 murine macrophages. AB - Whole plant methanolic extracts of 14 traditionally used medicinal herbs were evaluated for their anti-inflammatory activity. Extracts of Grindelia robusta, Salix nigra, Arnica montana, and Quassia amara showed up to 4.5-fold inhibition of nitric oxide (NO) production in the J774 murine macrophage cells challenged with LPS without cytotoxicity. These four selected extracts significantly reduced the protein levels of inducible NO synthase (iNOS) and the cyclooxygenase-2 (COX 2) as observed by Western blot analysis. Culture supernatants from cells treated with these extracts indicated 3-5-fold reduction of tumor necrosis factor-alpha (TNF-alpha). However, only G. robusta and Q. amara extracts significantly inhibited (by 50%) IL-1beta and IL-12 secretions. Furthermore, all these plant extracts were shown to prevent the LPS-mediated nuclear translocation of nuclear factor-kappaB (NF-kappaB). All the above observations indicate the anti inflammatory potential of these plant extracts. PMID- 19813080 TI - Simultaneous determination of ritodrine and isoxsuprine using coupling technique of synchronous fluorimetry and H-point standard addition method. AB - Simultaneous determination of two structurally related ss(2) adrenergic receptor agonists namely, Ritodrine HCl (RTH) and Isoxsuprine HCl (ISP) was performed using coupling technique of synchronous fluorimetry and H-point standard addition method. Under optimum conditions, linear determination ranges were 1.48 - 14.80 x 10(-6) mol L(-1) and 1.54 - 15.44 x 10(-6) mol L(-1) for ISP and RTH respectively. RTH and ISP could be determined simultaneously without interference from each other when their concentration ratio varies from 5:1 to 1:5 in the mixed sample. The proposed method was applied to the determination of RTH and ISP in synthetic mixture of pharmaceutical samples, the accuracy and precision of the results were satisfactory. PMID- 19813082 TI - Calpain in atrial fibrillation: friend or foe?: editorial to: "anti-apoptotic effects of a calpain inhibitor on cardiomyocytes in a canine rapid atrial fibrillation model" by Yue Li et al. PMID- 19813081 TI - Decisions to seek healthcare based on family health history among urban Appalachian women. AB - Family health history (FHH) is a valuable health promotion tool that can be used to assess disease risk and make lifestyle and screening recommendations. However, few FHH resources exist for medically underserved populations such as the urban Appalachian community in Cincinnati Ohio. Women of Appalachian heritage with less than a college education who did and did not participate in a prior FHH education session were interviewed by phone in a semi-structured format. Interviewees were asked to discuss their understanding of FHH and the role FHH played in seeking (or not seeking) medical care. Analysis of their discussion identified four overarching themes as well as a model identifying conditions that facilitated or impeded the choice to seek medical care based on FHH. Findings from this study may be used to develop targeted FHH educational interventions in the urban Appalachian and other communities. PMID- 19813083 TI - The impact of a community-based pilot health education intervention for older people as caregivers of orphaned and sick children as a result of HIV and AIDS in South Africa. AB - The increasing HIV and AIDS epidemic in South Africa poses a substantial burden to older people, in particular older women who mainly provide care for sick adult children and their grandchildren who have become orphaned and rendered vulnerable by the death or illness of their parents. In this study, 202 isiXhosa speaking older caregivers from Motherwell in the Eastern Cape Province of South Africa were trained to provide care for grandchildren and adult children living with HIV or AIDS. Based on a community needs assessment, a health education intervention comprising four modules was designed to improve skills and knowledge which would be used to assist older people in their care-giving tasks. Some topics were HIV and AIDS knowledge, effective intergenerational communication, providing home based basic nursing care, accessing social services and grants, and relaxation techniques. Structured one-on-one interviews measured differences between pre intervention and post-intervention scores among those who attended all four modules vs. those that missed one or more of the sessions. The results demonstrated that older people who participated in all four workshops perceived themselves more able and in control to provide nursing care. The participants also showed a more positive attitude towards people living with HIV or AIDS and reported an increased level of HIV and AIDS knowledge. The results provided valuable information upon which the development of future interventions may be based and psychosocial and structural needs of the older caregivers may be addressed by relevant stakeholders. PMID- 19813084 TI - Genetic and environmental influences on risky sexual behaviour and its relationship with personality. AB - Risky sexual behaviour is a major health issue in society, and it is therefore important to understand factors that may predispose individuals to such behaviour. Research suggests a link between risky sexual behaviour and personality, but the basis of this link remains unknown. Hans Eysenck proposed that personality is related to sexual behaviour via biological underpinnings of both. Here we test the viability of this perspective by analysing data from identical and non-identical twins (N = 4,904) who completed a questionnaire assessing sexual attitudes and behaviour as well as personality. Using genetic modelling of the twin data, we found that risky sexual behaviour was significantly positively correlated with Impulsivity (r = .27), Extraversion (r = .24), Psychoticism (r = .20), and Neuroticism (r = .09), and that in each case the correlation was due primarily to overlapping genetic influences. These findings suggest that the genetic influences that shape our personality may also predispose us to risky sexual behaviour. PMID- 19813085 TI - Patterns and predictors of restrictive health care service use by youths with bipolar disorder. AB - Patterns of behavioral health service utilization were examined among youth diagnosed with bipolar disorder (n = 832). Youth were categorized as high, moderate, and low restrictive service users (43, 7, and 50%, respectively). Factors associated with receiving care in highly restrictive settings included: numerous co-occurring diagnoses and being enrolled in managed care. Youth with regular outpatient visits were less likely to receive care in highly restrictive settings. This analysis provides a broad and dramatic picture of the intensity of services needed by most youth with bipolar. Having regular outpatient services shows promise with regard to reducing costly care in restrictive settings. PMID- 19813086 TI - Vigilance and avoidance of threat in the eye movements of children with separation anxiety disorder. AB - The vigilance-avoidance attention pattern is found in anxious adults, who initially gaze more at threatening pictures than nonanxious adults (vigilance), but subsequently gaze less at them than nonanxious adults (avoidance). The present research, using eye tracking methodology, tested whether anxious children show the same pattern. Children with separation anxiety disorder or no mental disorder viewed pairs of pictures, while the direction of their gaze was tracked. Each picture pair showed one picture of a woman separating from a child, the other picture of a woman reuniting with a child. The results supported the vigilance-avoidance model in children. Although the two groups' gaze direction did not differ during the first second of viewing, anxious children gazed significantly more at separating (threatening) pictures than nonanxious children after a period of 1 s. But after 3 s the pattern reversed: anxious children gazed significantly less at the separating pictures than nonanxious children. PMID- 19813087 TI - Stable expression of neurogenin 1 induces LGR5, a novel stem cell marker, in an immortalized human neural stem cell line HB1.F3. AB - Neural stem cells (NSC) with self-renewal and multipotent properties serve as an ideal cell source for transplantation to treat spinal cord injury, stroke, and neurodegenerative diseases. To efficiently induce neuronal lineage cells from NSC for neuron replacement therapy, we should clarify the intrinsic genetic programs involved in a time- and place-specific regulation of human NSC differentiation. Recently, we established an immortalized human NSC clone HB1.F3 to provide an unlimited NSC source applicable to genetic manipulation for cell-based therapy. To investigate a role of neurogenin 1 (Ngn1), a proneural basic helix-loop-helix (bHLH) transcription factor, in human NSC differentiation, we established a clone derived from F3 stably overexpressing Ngn1. Genome-wide gene expression profiling identified 250 upregulated genes and 338 downregulated genes in Ngn1 overexpressing F3 cells (F3-Ngn1) versus wild-type F3 cells (F3-WT). Notably, leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5), a novel stem cell marker, showed an 167-fold increase in F3-Ngn1, although transient overexpression of Ngn1 did not induce upregulation of LGR5, suggesting that LGR5 is not a direct transcriptional target of Ngn1. KeyMolnet, a bioinformatics tool for analyzing molecular relations on a comprehensive knowledgebase, suggests that the molecular network of differentially expressed genes involves the complex interaction of networks regulated by multiple transcription factors. Gene ontology (GO) terms of development and morphogenesis are enriched in upregulated genes, while those of extracellular matrix and adhesion are enriched in downregulated genes. These results suggest that stable expression of a single gene Ngn1 in F3 cells induces not simply neurogenic but multifunctional changes that potentially affect the differentiation of human NSC via a reorganization of complex gene regulatory networks. PMID- 19813088 TI - FoxM1 down-regulation leads to inhibition of proliferation, migration and invasion of breast cancer cells through the modulation of extra-cellular matrix degrading factors. AB - Forkhead box M1 (FoxM1) transcription factor is known to play important role in human cancers which, in part, is mediated by its ability to modulate cell cycle regulatory proteins as well as genes involved in cell proliferation and differentiation. In breast cancer, FoxM1 down-regulation is increasingly being recognized as an important mechanism for the targeted activity of anti-cancer agents. However, the mechanistic insight in support of the role of FoxM1 in aggressive breast cancer is poorly understood. We have tested the biological consequence of FoxM1 down-regulation and up-regulation in breast cancer cell lines and found that the down-regulation of FoxM1 in MDA-MB-231 and SUM149 cells by siRNA approach inhibited cell growth, clonogenicity, migration, and invasion. We also found decreased expression of CDK2 and E2F1 with concomitant increase in p21 and p27 proteins, suggesting an important role of FoxM1 in cell cycle progression. In contrast, over-expression of FoxM1 by cDNA transfection, in breast cancer cells (SUM102 and SKBR3) expressing low levels of FoxM1, resulted in increased cell proliferation, migration, and invasion. Moreover, down regulation of FoxM1 inhibited the expression of many factors that are involved in the degradation of extra cellular matrix and angiogenesis such as uPA, uPAR, MMP 2, MMP-9, and vascular endothelial growth factor (VEGF) as well as inhibited the activity of MMP-9 and VEGF. Interestingly, over-expression of uPA by cDNA transfection abrogated the cellular effects that were observed by the down regulation of FoxM1. Taken together, these results suggest the potential application of FoxM1 down-regulation as a novel approach for the treatment of aggressive breast cancer. PMID- 19813089 TI - A general definition of influence between stochastic processes. AB - We extend the study of weak local conditional independence (WCLI) based on a measurability condition made by (Commenges and Gegout-Petit J R Stat Soc B 71:1 18) to a larger class of processes that we call D'. We also give a definition related to the same concept based on certain likelihood processes, using the Girsanov theorem. Under certain conditions, the two definitions coincide on D'. These results may be used in causal models in that we define what may be the largest class of processes in which influences of one component of a stochastic process on another can be described without ambiguity. From WCLI we can construct a concept of strong local conditional independence (SCLI). When WCLI does not hold, there is a direct influence while when SCLI does not hold there is direct or indirect influence. We investigate whether WCLI and SCLI can be defined via conventional independence conditions and find that this is the case for the latter but not for the former. Finally we recall that causal interpretation does not follow from mere mathematical definitions, but requires working with a good system and with the true probability. PMID- 19813090 TI - Patterns of cardiovascular control during repeated tests of orthostatic loading. AB - To investigate patterns of cardiovascular control, a protocol of head up tilt (HUT) followed by lower body negative pressure (LBNP), which represents a significant cardiovascular control challenge, was employed. Linear regression of beat-to-beat heart rate (HR) and mean blood pressure (MBP) data collected over repeated tests was used to analyze control response during the LBNP phase of the combined HUT + LBNP protocol. Four runs for each of 10 healthy young males reaching presyncope were analyzed. Subjects were classified into 2 groups based on the consistency of MBP regulation in response to central hypovolemia induced by LBNP. The consistent group tended to exhibit consistent HR slope (rate of change of HR over time as calculated by linear regression) whereas subjects in the inconsistent group could not be easily classified. Subjects with consistent MBP maintenance exhibited patterns suggesting a consistency of response in cardiovascular control whereas subjects less successful in maintaining MBP exhibited less clearly defined patterns over four runs. PMID- 19813091 TI - A servo-controlled canine model of stable severe ischemic left ventricular failure. AB - Reversible left ventricular failure was produced in conscious dogs by compromise of the coronary circulation. In animals with prior left anterior descending coronary artery occlusion, mean left atrial pressure (LAP) was incorporated into an automatic feedback control system used to inflate a balloon cuff on the circumflex (Cfx) coronary artery. The system could produce stable increases in LAP to 15-20 mm Hg. The dominating system transfer function was the ratio of LAP to balloon volume (BV), which was characterized by a fixed delay (5 s), with LAP/BV = (8e(-jomegatau ))/(0.02 + jomega). The system was stabilized by a phase lead network to reduce oscillations of LAP. A total of seven experiments were conducted in three dogs, and testing of inotropic agents was possible in three experiments under stable conditions with the pump off after an hour or more of operation. Problems encountered were 0.003-0.008 Hz oscillations in LAP in three experiments, which could usually be controlled by reducing the system gain. Late stage ventricular fibrillation occurred in all three animals, but defibrillation was easily accomplished after deflating the Cfx balloon. This system produces reversible left ventricular failure solely due to ischemia, thus closely simulating clinical heart failure due to coronary insufficiency. PMID- 19813092 TI - Legal consequences of kleptomania. AB - Although studies have examined clinical characteristics of kleptomania, no previous studies have examined the legal consequences of kleptomania. From 2001 to 2007, 101 adult subjects (n = 27 [26.7%] males) with DSM-IV kleptomania were assessed on sociodemographics and clinical characteristics including symptom severity, comorbidity, and legal repercussions. Of 101 subjects with kleptomania, 73.3% were female. Mean age of shoplifting onset was 19.4 +/- 12.0 years, and subjects shoplifted a mean of 8.2 +/- 11.0 years prior to meeting full criteria for kleptomania. Co-occurring depressive, substance use, and impulse control disorders were common. Sixty-nine subjects with kleptomania (68.3%) had been arrested, 36.6% had been arrested but not convicted, 20.8% had been convicted and incarcerated after conviction, while only 10.9% had been convicted and not incarcerated after conviction. Kleptomania is associated with significant legal repercussions. The findings emphasize the need for rigorous treatment approaches to target kleptomania symptoms and prevent re-offending. PMID- 19813094 TI - Ethical aspects of the use of stem cell derived gametes for reproduction. AB - A lot of interest has been generated by the possibility of deriving gametes from embryonic stem cells and bone marrow stem cells. These stem cell derived gametes may become useful for research and for the treatment of infertility. In this article we consider prospectively the ethical issues that will arise if stem cell derived gametes are used in the clinic, making a distinction between concerns that only apply to embryonic stem cell derived gametes and concerns that are also relevant for gametes derived from adult stem cells. At present, it appears preferable to use non-embryonic stem cells for the derivation of gametes. Adult stem cell derived gametes do not present any problems with regard to the moral status of the human embryo, bypass the safety risks linked to SCNT and do not present any ambiguity or novel problems with regard to informed consent, psychological consequences for the child or genetic parenthood. A remaining ethical concern, however, regards the safety of the procedure in terms of the welfare of the resulting children. This should spark a thorough reflection on how far one must go to accommodate a person's wish to have a genetically related child. PMID- 19813093 TI - Implicit learning in aging: extant patterns and new directions. AB - Research suggests that the striatum plays an important role in implicit learning (IL). The striatum exhibits marked age-related morphological and neurochemical losses. Yet, behavioral studies suggest that IL is generally well preserved in old age, and that age-related differences emerge only when highly complex IL tasks are used. In this review, we integrate behavioral and neuroimaging evidence on IL in aging. We suggest that relative stability of IL in old age may reflect neural reorganization that compensates for age-related losses in striatal functions. Specifically, there may be an age-related increase in reliance on extrastriatal regions (e.g., medial-temporal, frontal) during IL. This reorganization of function may be beneficial under less taxing performance conditions, but not when task demands become more challenging. PMID- 19813095 TI - Aesthetic, emotion and empathetic imagination: beyond innovation to creativity in the health and social care workforce. AB - The Creativity in Health and Care Workshops programme was a series of investigative workshops aimed at interrogating the subject of creativity with an over-arching objective of extending the understanding of the problems and possibilities of applying creativity within the health and care sector workforce. Included in the workshops was a concept analysis, which attempted to gain clearer understanding of creativity and innovation within this context. The analysis led to emergent theory regarding the central importance of aesthetics, emotion and empathetic imagination to the generation of creative and innovative outcomes that have the capacity to promote wellbeing in the health and social care workforce. Drawing on expertise in the field, this paper outlines the concept analysis and subsequent reflection. PMID- 19813096 TI - From the editor. PMID- 19813097 TI - Preimplantation genetic diagnosis for alpha-thalassaemia in China. AB - PURPOSE: To report the usage of PGD for alpha-thalassaemia with the - -(SEA) genotype. METHOD: A PGD protocol using fluorescent gap PCR was performed for 51 cycles on 43 couples with the - -(SEA) genotype. Allele drop-out and amplification failure rates were retrospectively analyzed. RESULTS: A total of 472 embryos were biopsied. Amplification was achieved in 390 blastomeres, accounting for an amplification rate of 82.6%. In total, 120 wild-type, 94 heterozygotes and 140 homozygous mutant embryos were diagnosed. The successful diagnosis rate was 75.0%. The ADO rate in 49 blastomeres from six donated embryos was 16.4%. One hundred and fifty four embryos were transferred, resulting in 25 clinical pregnancies with an implantation rate of 24.0%. CONCLUSIONS: Single round fluorescent gap PCR is a feasible and effective strategy in the PGD for alpha-thalassaemia with the - -(SEA) genotype. PMID- 19813098 TI - Influence of smoking on outcome of COH and IUI in subfertile couples. AB - AIM: To evaluate the influence of smoking on the outcome of COH and IUI in subfertile couples. PATIENTS AND METHODS: We reviewed the medical files of all consecutive women, age 14 mm in diameter, or E2 levels on the day of hCG administration. CONCLUSION: Smokers undergoing COH with IUI required a significantly higher gonadotropin dosage than nonsmokers in order to achieved a comparable pregnancy rate. PMID- 19813100 TI - Empirical Bayes and semi-Bayes adjustments for a vast number of estimations. AB - Investigators in modern molecular/genetic epidemiology studies commonly analyze data on a vast number of candidate genetic markers. In such situations, rather than conventional estimation of effects (odds ratios), more accurate estimation methods are needed. The author proposes consideration of empirical Bayes and semi Bayes methods, which yield 'adjustments for multiple estimations' by shrinking conventional effect estimates towards the overall average effect. PMID- 19813101 TI - A new beta-glucosidase gene from the zygomycete fungus Rhizomucor miehei. AB - In this study, a beta-glucosidase coding gene (bgl) of the zygomycete fungus Rhizomucor miehei has been cloned and characterized. The gene comprises a total of 2,826 bp including the coding sequence of a 717 amino acids length putative protein and 10 introns dispersed in the whole coding region. The putative N-and C terminal catalytic domains (aa 68 to aa 274 and aa 358-601, respectively) were identified; the two domains are connected with a 84-amino-acids linker. The catalytic region showed an extensive sequence homology with other fungal beta glucosidases classified as family 3 glycoside hydrolases. The isolated Rhizomucor gene was expressed in the related fungus Mucor circinelloides. Transformant Mucor strains maintained the introduced plasmid in an autoreplicative manner and showed significantly higher cellobiase activity than the recipient strain. PMID- 19813102 TI - Heterologous expression, purification and characterization of the influenza A virus M2e gene fused to Mycobacterium tuberculosis HSP70(359-610) in prokaryotic system as a fusion protein. AB - One of the concerns about influenza A vaccine based on M2e protein is their limited potency; hence, optimal approaches to enhance immunogenicity of M2e protein immunization remain to be established. It seems by linking this M2e peptide to an appropriate carrier such as mycobacterium tuberculosis C-terminal 28-kDa domain of HSP70 (HSP70(359-610)), we can render it very immunogenic. According to previous reports, this study was designed to produce a novel influenza A virus recombinant fusion protein consisted of M2e, a potent immunogenic protein from influenza A virus, fused to C-terminal domain of mycobacterium tuberculosis HSP70, HSP70(359-610), as a carrier and adjuvant. We fused the genes of M2e and HSP70 ( 359-610 ) then inserted in pQE-60, prokaryotic expression vector. This recombinant fusion protein with a 6xHis-tag was successfully over expressed in Escherichia coli M-15. The recombinant fusion protein was purified by Ni-NTA affinity chromatography under denaturing conditions, followed by urea gradient dialysis. The purified fusion protein was analyzed on SDS-PAGE. Western blot assay was used to examine the immunoreaction of the expressed protein using commercial penta-His HRP conjugate antibody. The antigenicity and biological activity of the recombinant protein was also qualitatively detected on the infected MDCK cells surface by immunofluorescence and cell-ELISA assay using rabbit's immunized antiserum. This observation suggest that the expressed fusion protein is useful as a universal recombinant vaccine for overcoming highly mutational influenza virus, but more immunological study in animal lab remains to be evaluated. PMID- 19813103 TI - The association of body mass index and health-related quality of life in the general population: data from the 2003 Health Survey of England. AB - OBJECTIVES: The link between obesity/overweight and life-threatening illnesses is well established. The objective of this study was to investigate the relationship between body mass index (BMI) and health-related quality of life (HRQoL), and any differences between men and women, in the general population of England. METHODS: HRQoL data (from EQ-5D responses of 14,416 individuals aged >or=18 in the 2003 Health Survey for England) were used, and linear regression analyses were conducted to examine the relationship between BMI and HRQoL. RESULTS: A significant association between BMI and HRQoL was found after controlling for factors such as gender, age, and obesity-related comorbidities. The maximum HRQoL was reached at a BMI of 26.0 in men and 24.5 in women, demonstrating that BMI is negatively associated with HRQoL for both underweight and obese individuals. At higher BMI values, men reported higher HRQoL than women; at lower BMI values, HRQoL was lower in men than women. CONCLUSIONS: There is a significant association between BMI and HRQoL in men and women in the general population. Nearly all aspects of HRQoL are adversely affected by elevated BMI. PMID- 19813105 TI - Abstracts of the Neurocritical Care Society 7th Annual Meeting. November 11-14, 2009. New Orleans, Louisiana, USA. PMID- 19813104 TI - Prevalence and characterization of ECG abnormalities after intracerebral hemorrhage. AB - BACKGROUND: Although electrocardiographic (ECG) abnormalities are well known in ischemic stroke and subarachnoid hemorrhage, these changes have only rarely been investigated systematically in patients with intracerebral hemorrhage (ICH). The purpose of this study is to investigate the prevalence and type of ECG abnormalities in a consecutive series of ICH patients, and their possible association with pre-defined neurological and radiological parameters. METHODS: The study population consisted of all consecutive patients with non-traumatic, intraparenchymal ICH admitted to the Academic Medical Center (AMC) between January 1, 2007 and October 1, 2007. Baseline information was prospectively registered in the AMC Stroke Register. ECGs obtained within 2 days after the initial hemorrhage were analyzed by one blinded observer. Admission cranial CT scans were re-analyzed by two blinded observers. RESULTS: Thirty-one patients were included. Twenty-five patients (81%) had one or more ECG abnormalities. The most frequently observed ECG abnormality was QTc prolongation (36%), followed by ST-T morphologic changes (23%), sinus bradycardia (16%), and inverted T wave (16%). No patient was initially misdiagnosed for having myocardial ischemia. QTc prolongation was associated with ICH involvement of the insular cortex [OR 10.9 (95% CI 1.0-114.6)] and presence of intraventricular blood and hydrocephalus on admission CT scan [OR 10.8 (95% CI 1.6-70.9)]. CONCLUSIONS: In ICH patients ECG abnormalities are common. QTc prolongation seems associated with insular cortex involvement, with the presence of intraventricular blood, and with hydrocephalus. A larger cohort of continuously monitored ICH patients is necessary to investigate whether these ECG abnormalities are associated with poor outcome or death. PMID- 19813106 TI - One thing leads to another: GBS complicated by PRES and Takotsubo cardiomyopathy. AB - BACKGROUND: To describe a patient with Guillane-Barre syndrome (GBS), posterior reversible encephalopathy syndrome (PRES) and takotsubo cardiomyopathy, and in doing so, to postulate a new mechanism for hypotension in the setting of dysautonomia in GBS. METHODS: Case report. RESULTS: We report an 82-year-old woman who presented with bilateral lower extremity weakness and back pain following an upper respiratory viral illness. Within 4 days she became markedly hypertensive and developed status epilepticus. Brain MRI revealed patchy bilateral occipital lesions with T2 signal hyperintensity consistent with PRES. Her clinical exam, CSF, and electrophysiologic findings were consistent with GBS. She became relatively hypotensive and transthoracic echocardiogram showed severe apical akinesis. The patient clinically improved, and echocardiogram and MRI abnormalities resolved within 2 weeks. CONCLUSIONS: The autonomic effects of GBS may cause a variety of reversible clinical syndromes associated with sympathetic dysfunction including PRES and takotsubo cardiomyopathy; both of which are self limited. Relative hypotension in GBS may be caused in part by neurogenic stunned myocardium. PMID- 19813107 TI - Loss of p16INK4, alone and with overexpression of osteopontin, correlates with survival of patients with spinal metastasis from hepatocellular carcinoma. AB - The survival of patients with hepatocellular carcinoma (HCC) has been improved with various diagnostic tools and treatment modalities. Consequently, spinal metastases from HCC are diagnosed more frequently. We investigated the clinical biomarkers of HCC patients presenting with spinal metastasis. Between January 2001 and December 2007, we recruited 30 consecutive HCC patients presenting with spinal metastasis. Their tissue samples were collected and analyzed by immunohistochemistry in a tissue microarray. A total of 16 proteins were assessed in the tissue microarray; we found that expression of p16(INK4) correlated with the survival time (log-rank test, P = 0.05), and loss of p16(INK4) was significantly associated with osteopontin overexpression (Fisher exact test: P = 0.045, logistic regression: P = 0.024, OR = 0.184, 95% CI 0.035-0.963). Patients with osteopontin (-) and with p16(INK4) (+) lived longer than patients with osteopontin (+) and with p16(INK4) (-). We found that p16(INK4) and osteopontin might be the biomarkers of patients with spinal metastasis from HCC, a more large scaled randomized study might be required to confirm the result and study the mechanism. PMID- 19813108 TI - Substance use disorders and risk for completed suicide. AB - Substance use disorders are among the most frequent psychiatric disorders found in suicides. In psychological autopsy studies between 19% and 63% of all suicides suffered from substance use disorders, mostly from alcohol use disorders. Suicide risk is highly increased in substance use disorders, particularly in alcohol use disorders, and in co-morbid alcoholism and depression. So far, some risk factors for suicide have been identified in alcoholism. Nevertheless, various questions about the relationship between substance use disorders and suicide remain open, which indicate directions for future research. PMID- 19813109 TI - Rates of non-suicidal self-injury in high school students across five years. AB - It is believed that the rate of non-suicidal self-injury (NSSI) among high school students is increasing. However, no known research has examined the rates of NSSI among non-clinical, high school samples of adolescents across several years. The current study utilized archival data from five academic years [2001 to 2005 (N = 1393)] to address this limitation by examining the lifetime prevalence rates of NSSI in urban high school students within the Midwestern United States. Results indicate a significant difference in rates across years by gender. Females showed a significant increase in their rates of NSSI compared to males, who showed a significant decline, during the last 3 years of the study. Implications of these findings for school personnel and directions for future research discussed. PMID- 19813110 TI - Role of IL-10 -1082, IFN-gamma +874, and TNF-alpha -308 genes polymorphisms in suicidal behavior. AB - In this study, it was determined whether the IL-10 -1082, IFN-gamma +874, and TNF alpha -308 polymorphisms were associated with suicidal behavior. One hundred forty five patients with suicidal behavior and 160 normal individuals were genotyped for IL-10 -1082, IFN-gamma +874, and TNF-alpha -308 polymorphisms using ASO-PCR method. TNF-alpha -308 G/G genotype has been increased in males with completed suicide behavior versus control group (p value = 0.017). IL-10 -1082 A/A genotype is higher in both male and female suicide completed groups (p value = 0.017). IFN-gamma (+874) A/A genotype was significantly higher in males with completed suicide behavior versus normal male control (p value = 0.027). It can be concluded that IL-10, IFN-gamma, and TNF-alpha polymorphisms may play a role in suicidal behavior. PMID- 19813111 TI - Cannabis use and deliberate self-harm in adolescence: a comparative analysis of associations in England and Norway. AB - The objective of this study was to test hypotheses on causality and selection regarding associations between cannabis use and deliberate self-harm (DSH) among adolescents. School surveys were conducted among 9,800 adolescents in England and Norway applying identical measures on deliberate self-harm, suicidal thoughts, cannabis use, and various potential confounders. Cannabis use was more prevalent in England than in Norway. It was associated with DHS, suicidal thoughts and various risk factors for DSH. However, these associations were stronger in Norway than in England. The adjusted associations between cannabis use and suicidal thoughts were non-significant in both countries. The adjusted cannabis-DSH association was non-significant in England but significant in Norway. Elevated risk of DSH in adolescent cannabis users seems to be mainly due to selection mechanisms. Thus the association is not likely to be direct but due to other shared contributory factors. PMID- 19813112 TI - Variation in RGS2 is associated with suicidal ideation in an epidemiological study of adults exposed to the 2004 Florida hurricanes. AB - This study examined whether rs4606, a single nucleotide polymorphism (SNP) in the translated region at the 3' end of RGS2, was related to suicidal ideation in an epidemiologic sample of adults living in areas affected by the 2004 Florida hurricanes. An epidemiologic sample of residents of Florida was recruited via random digit-dial procedures after the 2004 Florida hurricanes; participants were interviewed about suicidal ideation, hurricane exposure, and social support. Participants who returned buccal DNA samples via mail (n = 607) were included here. Rs4606 in RGS2 was associated with increased symptoms of current suicidal ideation (p < 0.01). Each "C" allele was associated with 5.59 times increased risk of having current ideation. No gene-by-environment interactions were found, perhaps due to low power. RGS2 rs4606 is related to risk of current suicidal ideation in stressor-exposed adults. PMID- 19813113 TI - The role of Expressed Emotion, Self-concept, Coping, and Depression in parasuicidal behavior: a follow-up study. AB - The objective of this study was to characterize Expressed Emotion in families of individuals with parasuicidal behavior, a non-fatal act in which there is self harm or deliberate excessive ingestion of a substance and to examine the significant relationships between Expressed Emotion (EE), Coping, Depression, Self-concept, and parasuicidal behaviors. The sample consisted of 67 subjects divided into two groups. The first group was made up of 34 parasuicides. Parasuicidal behavior, self-concept, coping, depression and the family's EE were assessed in this group. The control group was composed of 33 young people with identical characteristics (age: 15-24; gender: more female; and residence). All the subjects were followed up for a 9 month period, during which time EE and recurrent parasuicidal behaviors were assessed. Parasuicides showed significant differences (at a 0.05% significance level) in comparison to the control group They showed more Depressed, less Coping, and less Self-concept. There was also an intimate family atmosphere with a high EE. The parasuicides whose families had a high EE showed more recurrent parasuicidal behaviors. The results demonstrate that EE is a predictor of recurrent parasuicidal behaviors and that its assessment in families of young parasuicides is useful. PMID- 19813114 TI - Suicide attempts by poisoning in Hanoi, Vietnam: methods used, mental problems, and history of mental health care. AB - The objective of this paper was to investigate methods of poisoning, presence of mental problems, and the rate of psychiatric care among suicide attempters in Vietnam. Three hundred and nine suicide attempters by poisoning admitted to Bach Mai Hospital's Poison Control Center were investigated by trained nurses and qualified psychiatrists. Standardized questionnaires were used to assess methods of suicide, presence of mental problems, and psychiatric care. The most common methods were poisoning by psychotropic drugs in urban, and pesticides in rural areas. ICD-10 confirmed disorders were present in 68% of the cases and 73% rated positive on SRQ-20. Most patients were not in contact with psychiatric care. Restrictions on availability and handling of drugs and pesticides should be reinforced. Better infrastructures are needed for identification and treatment of persons with mental disorders. PMID- 19813115 TI - Family environment and suicidal ideation among bipolar youth. AB - The objective of this study was to examine the association between family environment and suicidal ideation among youth with bipolar disorder. Subjects included 446 bipolar (BP) youth (age 7-17) enrolled in the Course and Outcome of Bipolar Youth study. Current suicidal ideation, family functioning and family stress were assessed at intake. BP youth with current suicidal ideation reported more conflict with their mother and less family adaptability. Ideators endorsed more stressful family events over the prior year and higher rates of specific familial stressors. Clinicians treating bipolar youth should consider family stress when conducting suicide risk assessment. Treatment goals may include enhancing family communication and addressing issues of loss. PMID- 19813116 TI - A nonverbal learning disability in a case of central hypoventilation syndrome without a PHOX2B gene mutation. AB - This study examines the neuropsychological profile of a boy with congenital central hypoventilation syndrome (CCHS) without a paired-like homeobox gene (PHOX2B) mutation. CCHS is a rare disorder of autonomic nervous system development characterized by an impaired ventilatory response to hypercarbia and hypoxemia. Mild intellectual deficits are common but a specific cognitive profile is not established in CCHS. We describe a nonverbal learning disorder as a CCHS endophenotype and recommend that detailed neuropsychological testing be performed on all individuals with CCHS. Defining the psycho-educational needs in CCHS may avert compounding the emotional and medical stresses of this already debilitating disorder. PMID- 19813117 TI - Making 'good girls': sexual agency in the sexuality education of low-income black girls. AB - Critics argue that abstinence-only programmes reinforce gender inequality when they contain discourses that equate being a 'good girl' with sexual restraint. Yet they too often overlook how racial and class inequalities shape discourses about girls' sexual agency. This ethnography extends gender scholarship by analysing the racialised, classed and gendered dynamics of an abstinence-only programme for low-income black girls. It finds that black adults viewed the girls as sexually vulnerable because of racism and class inequality. They tried to mediate this vulnerability by transforming girls into sexual agents. They did so, though, by exaggerating the gendered discourses of the official sexuality education curricula that framed girls as victims and their sexual restraint as a matter of morality. Thus, the programme reinforced gender inequality while trying to disrupt race and class inequalities. PMID- 19813118 TI - Sexual health knowledge, sexual relationships and condom use among male trekking guides in Nepal: a qualitative study. AB - People in Nepal generally hold fairly traditional views about sex and sexual health, whilst Western tourists often have a more liberal approach towards sex and relationships. There is evidence that significant sexual interaction occurs between male trekking guides and female travellers and/or local female sex workers in Nepal. This qualitative study explored trekking guides' sexual health knowledge, sexual relationships and condom use with female trekkers and local female sex workers. A total of 21 in-depth interviews were conducted with male trekking guides. Most reported having had sexual relationships with female trekkers and local female sex workers. Explanations for intercourse with female trekkers included: financial support; getting future trekkers through word-of mouth advertising from the women they have had sex with; and opportunities for emigration. Interestingly, sexual intercourse is reported as more likely to be initiated by female trekkers than by guides, and more so by older women. In contrast, the main reasons for having sex with local female sex workers included: romantic love or sexual excitement and novelty. Awareness regarding sexual health was high among guides, but several factors discouraged the regular use of condoms. Further research with female tourists would help understand the motivations and reasons for their sexual behaviour. PMID- 19813119 TI - Quality of offspring-The impact of new reproductive technologies in Asia. Editorial introduction. PMID- 19813120 TI - Gendered talk about sex, sexual relationships and HIV among young people in Papua New Guinea. AB - This paper presents findings from a qualitative study carried out in three secondary schools in Eastern Highlands Province, Papua New Guinea (PNG). Seventy three Year 12 students took part in eight gender-specific focus group discussions (three female and five male). Irrespective of gender, respondents predominately understood sex as being for the sole purpose of reproduction within marriage. When discussing sex and sexual relationships, young men used explicit language and referred specifically to sexual organs and activities. Young women did not. Less concerned for privacy, young men talked in public spaces and in groups with same-sex peers about sex and sexual expression, whereas young women discussed such matters one-on-one and in private. These gender differences provide useful entry points for developing appropriate sex and HIV education programmes involving young people in PNG. PMID- 19813121 TI - Nonprofit health care services marketing: persuasive messages based on multidimensional concept mapping and direct magnitude estimation. AB - Persuasive messages for marketing healthcare services in general and coordinated care in particular are more important now for providers, hospitals, and third party payers than ever before. The combination of measurement-based information and creativity may be among the most critical factors in reaching markets or expanding markets. The research presented here provides an approach to marketing coordinated care services which allows healthcare managers to plan persuasive messages given the market conditions they face. Using market respondents' thinking about product attributes combined with distance measurement between pairs of product attributes, a conceptual marketing map is presented and applied to advertising, message copy, and delivery. The data reported here are representative of the potential caregivers for which the messages are intended. Results are described with implications for application to coordinated care services. Theory building and marketing practice are discussed in the light of findings and methodology. PMID- 19813122 TI - The path of patient loyalty and the role of doctor reputation. AB - Patient loyalty to doctors is relevant to medical services in which doctor patient relationships are central and for which competition has increased in recent years. This study aims at understanding the process whereby patients develop loyalty to their doctor and doctor reputation has a moderating role. Based on a randomization of subjects, the study offers and tests an explanation chain representing key variables determining patient loyalty: patient commitment, trust and satisfaction, and doctor reputation. Primary data was collected using a structured questionnaire from a quota sample of regular patients in a large city in South America. The patients most committed to their doctor are more loyal to them. In turn, commitment is determined by patient trust, which is determined by patient satisfaction. Doctor reputation positively influences both patient trust and satisfaction. The explanation chain not only gives an account of how patient loyalty is formed; it also identifies a path health professionals can follow to secure patient loyalty. PMID- 19813123 TI - Healthcare cost and predictive factors: high- and low-utilization model development. AB - Healthcare costs in the U.S. are the highest in the world and are increasing rapidly. With increasing costs, employers and health insurance companies are trying to contain the cost of healthcare. This study aims at developing a predictive model relating enrollees' healthcare insurance claims to their health risks while controlling for gender, age, and their previous year's claims. Our study findings suggest that age, gender, and a previous year's healthcare expenditure are strong predictors of healthcare cost and that none of the six biomarkers was a significant predictor. Even though these biomarkers failed to contribute to the predictive model, they are, nonetheless, important predictors of future chronic diseases, many of which are leading causes of death in the U.S. PMID- 19813124 TI - The impact of hospital quality on profits, volume, and length of stay. AB - Using 2006-2007 data from a sample of 6,082 U.S. hospitals, quality of care measures developed by the Hospital Quality Alliance (HQA) are applied to heart attack, heart failure, pneumonia, and surgery. Quality of care for these was related to both higher profits per case and increased number of cases. Length of stay was inversely related to quality of care for preventative (surgical and heart attack) care. PMID- 19813125 TI - Do physicians' beliefs about genetic engineering influence their likelihood of prescribing a biopharmaceutical? An empirical investigation. AB - Biopharmaceuticals present one of the fastest growing segments of the pharmaceutical industry. Biopharmaceuticals are produced through genetic engineering, a technology that some believe is unethical and can have unforeseen consequences to individual health and the environment (Bredhal, 1999; Gaskell et al., 1999; Hallman et al., 2002). In this study, we investigate whether physicians' beliefs about genetic engineering have any influence on their likelihood of prescribing a biopharmaceutical. A sample of 175 physicians practicing in one state in the mid-south region of the U.S. was selected and their beliefs, importance of genetic modification as a drug attribute and the likelihood of prescribing a biopharmaceutical, were measured. Results showed that the physicians' beliefs about genetic engineering related to their likelihood of prescribing this class of drugs. In addition, the study also found that the importance of genetic modification as a drug attribute moderated the relationship between beliefs and likelihood in the decision making process. The implications of these findings are also discussed. PMID- 19813126 TI - External recommendations versus internal satisfaction in health care: a case study in India. AB - This study examines the relative roles of external recommendations and internal satisfaction in influencing patient loyalty to physicians in India. The results indicate that recommendations result in preliminary loyalty formation by the patient toward the physician, but only until the patient formulates individual loyalty based upon their own personal experiences. At that point, the significance of external sources of recommendation in the determination of patient loyalty becomes insignificant. Physicians should strive at fostering bonds of emotional attachment in their present patients so they become strongly loyal and spread positive word-of-mouth, which, in turn, may result in new patients to the physician. PMID- 19813127 TI - The illusion of weightlessness. AB - The ballet pointe shoe is an interesting example of a product that has developed from the experimentation of a dancer to extend the art form. Over time, as the use of such a shoe became part of the tradition the form stagnated and there was little formal interaction with the user as to its continued effectiveness in terms of injury, cost and fit. This work re-examines the shoe from the dancer's point of view and demonstrates a way forward that illustrates that even with severe constraints the designer with a user-centred approach can improve the performance of a product whilst maintaining a required external aesthetic. PMID- 19813128 TI - Sources of coding discrepancies in injury morbidity data: implications for injury surveillance. AB - The aim of this study is to examine the sources of coding discrepancy for injury morbidity data and explore the implications of these sources for injury surveillance. An on-site medical record review and recoding study was conducted for 4373 injury-related hospital admissions across Australia. Codes from the original dataset were compared with the recoded data to explore the reliability of coded data and sources of discrepancy. The most common reason for differences in coding overall was assigning the case to a different external cause category with 9.5% assigned to a different category. Differences in the specificity of codes assigned within a category accounted for 7.8% of coder difference. Differences in intent assignment accounted for 3.7% of the differences in code assignment. In the situation where 8% of cases are misclassified by major category, the setting of injury targets on the basis of extent of burden is a somewhat blunt instrument. Monitoring the effect of prevention programs aimed at reducing risk factors is not possible in datasets with this level of misclassification error in injury cause subcategories. Future research is needed to build the evidence base around the quality and utility of the International Statistical Classification of Diseases and Related Health Problems (ICD) classification system and the application of use of this for injury surveillance in the hospital environment. PMID- 19813129 TI - The contribution of executive functioning to academic achievement among male adolescents. AB - Recent factor analytic work suggests that the dimensions of conceptual flexibility, monitoring, and inhibition are distinguishable under the executive functioning (EF) umbrella. We examine relations between these constructs and performances on academic achievement tests among a sample of 11-16-year-old males (N = 151). EF contributed to the prediction of all academic domains beyond general intellectual functioning in distinct ways: Conceptual flexibility predicted reading and science, monitoring predicted reading and social studies, and inhibition predicted mathematics and science. These findings suggest that demands related to specific academic domains access different cognitive abilities and have implications for both intervention and research science. PMID- 19813130 TI - Working memory, response inhibition, and within-subject variability in children with attention-deficit/hyperactivity disorder or reading disorder. AB - This study compared children with ADHD (n = 19), reading disorder (RD; n = 17), ADHD+RD (n = 21), and control children (n = 19) on linguistic and executive function measures. We found no evidence of response inhibition problems in ADHD or RD when a baseline measure of functioning was taken into account. General working memory problems were only found in children with RD or ADHD+RD. Both children with ADHD and RD showed a highly inaccurate (more commission errors) and variable (higher within-subject standard deviation of reaction time) response style. The comorbid group made most errors, suggesting that different factors underlie the high error rate in both disorders. PMID- 19813131 TI - Advance care planning decisions of women with cancer: provider recognition and stability of choices. AB - Our prior research found that 43% of women with cancer relied on more than one person for advance care planning (ACP) and support. We conducted this follow-up study to address the stability of patient choices around ACP and providers' knowledge of patients' named supports. Living participants from the original survey were recontacted and asked to participate in this study. Of the original 215 participants, 113 (66%) participated. The median time between surveys was 23 months. At resurvey, 33 (26%) patients did not name the same person to all three roles. Controlling for age, race, partner status, tumor type, and remission status, naming one person for all three roles was associated with higher concordance at follow-up for primary support (PS) and health care proxy (HCP). Comparing patients' and providers' responses (N = 162), concordance was 71% for emergency contact (EC), 60% for PS, but only 51% for HCP. In this follow-up study, a smaller percentage of women named more than one person to the roles of EC, PS, and HCP compared to the original survey. Of concern, concordance between patient and provider was low, particularly for HCP. This was notable especially when responses changed over the interval time, and when ACP and PS roles were split between multiple people. Understanding the network of people who serve ACP and support roles may be important as we seek to improve the process of ACP and surrogate decision making. PMID- 19813132 TI - Investigation into the communication that takes place between nurses and patients during chemotherapy. AB - This study used the medium of videotaping to investigate the verbal and nonverbal communication that takes place between nurses and patients during chemotherapy administration. Eight chemotherapy sessions were video recorded and then analyzed for emerging themes. In addition, the videotapes were used in a reflective process with nurses. The findings showed that nurses were efficient in communicating about the physical and medical care of patients but were hesitant in exploring emotional issues. This study highlights the focus on physical symptoms and side effects by nurses in this setting. It also supports the need for further education and training in psychosocial assessment during chemotherapy administration. PMID- 19813133 TI - Cognitive and psychological factors associated with early posttreatment functional outcomes in breast cancer survivors. AB - Breast cancer survivors experience cognitive difficulties following chemotherapy, yet the effects of these deficits on functional outcomes have not been systematically evaluated. This study assessed the relationships between postchemotherapy cognitive difficulties and functional outcomes. Forty-six women with breast cancer were seen at 1-month postchemotherapy; data were collected on cognitive functioning, psychological variables, and physical symptoms. Wilcoxon signed-rank analyses revealed cognitive deficits in executive functioning and verbal fluency. Subsequent regression analyses demonstrated that poorer executive functioning was associated with decreased productivity, community involvement, and social role functioning. Poorer quality of life was predicted by depression and reluctance to seek social support, but not cognitive functioning. These findings indicate that executive functioning deficits are associated with important functional outcomes among breast cancer survivors 1-month postchemotherapy. Thus, treatment efforts should focus on addressing cognitive, as well as psychological and physical, issues among cancer survivors. PMID- 19813134 TI - The Cancer and Deity Questionnaire: a new religion and cancer measure. AB - We evaluated a new measure, the Cancer and Deity Questionnaire (CDQ), which assesses perceived relations with God after a cancer diagnosis. Based on object relations theory, the 12-item CDQ assesses benevolent and abandoning God representations. Sixty-one older participants with recent cancer diagnoses completed the questionnaire at baseline, and 52 of these participants completed the same questionnaire at follow-up. Internal consistency was excellent for the Benevolence scale (alpha = .97) and good for the Abandonment scale (alpha = .80). Moderate correlations with the Spiritual Well-Being Scale support divergent validity. Correlations between CDQ scales and the Styles of Religious Coping scales support convergent validity. The CDQ is brief, easily scored, practical for psycho-oncology research, and adaptable for use with other illnesses. PMID- 19813135 TI - The Getting-Out-of-Bed (GoB) scale: a measure of motivation and life outlook in older adults with cancer. AB - OBJECTIVE: To develop and evaluate the psychometric properties of a measure of motivation and life outlook (Getting-Out-of-Bed [GoB]). DESIGN: Secondary analysis of baseline and 6-month data from a longitudinal follow-up study of older breast cancer survivors. PARTICIPANTS: Women (N = 660) diagnosed with primary breast cancer stage I-IIIA disease, age >or=65 years, and permission to contact from an attending physician in four geographic regions in the United States (city-based Los Angeles, California; statewide in Minnesota, North Carolina, and Rhode Island). MEASUREMENT: Data were collected over 6-months of follow-up from consenting patients' medical records and telephone interviews with patients. Data collected included the 4-item GoB, health-related quality of life (HRQoL), breast cancer, sociodemographic, and health-related characteristics. RESULTS: Factor analysis produced, as hypothesized, one principal component with eigen values of 2.74(baseline) and 2.91(6-months) which explained 68.6%(baseline) and 72.7%(6-months) of total variance. In further psychometric analyses, GoB exhibited good construct validity (divergent: low nonstatistically significant correlations with unrelated constructs; convergent: moderate statistically significant correlations with related constructs; discriminant: distinguished high HRQoL groups with a high level of significance), excellent internal reliability (Cronbach's alpha 0.84(baseline), 0.87(6-months)), and produced stable measurements over 6-months. Women with GoB scores >or=50 at baseline were more likely at 6-months to have good HRQoL, good self-perceived health, and report regular exercise, indicating good predictive ability. CONCLUSION: GoB demonstrated overall good psychometric properties in this sample of older breast cancer survivors, suggestive of a promising tool for assessing motivation and life outlook in older adults. Nevertheless, because it was developed and initially evaluated in a select sample, using measures with similar but not exact content overlap further evaluation is needed before it can be recommended for widespread use. PMID- 19813136 TI - Religion and cancer: examining the possible connections. AB - Numerous sound scientific studies (cross-sectional and longitudinal) have found a positive correlation between religion and physical and mental health. In particular, there is evidence that demonstrates that religion helps cancer patients better adjust to and cope with their disease, at least psychologically. However, some research suggests that mediating factors associated with religion may explain the positive effects of religion on health. This article argues that even if this is the case, there is still intrinsic value to religion in that the mediators themselves are strongly connected to religion, and therefore religion is important to the patient in terms of coping, support, hope, and meaning. This has possible important implications for clinical practice. PMID- 19813137 TI - Physiological monitoring of the Olympic athlete. AB - As the winning margin in Olympic competition is so small, there is a continuous quest for improvements in the preparation of athletes at this standard. Therefore, even the smallest physiological improvements that result from modifications in training strategy, preparation regime or ergogenic aids are potentially useful. Unfortunately, there is a lack of research data on elite competitors, which limits our interpretation of current literature to the elite sporting environment. This places extra responsibility on the physiologist to carefully consider the most appropriate physiological variables to monitor, the best protocols to assess those variables, and the accurate interpretation of the test results. In this paper, we address the key issues of ecological validity, measurement error, and interpretation for the most commonly monitored physiological variables. Where appropriate, we also indicate areas that would benefit from further research. PMID- 19813138 TI - My face in yours: Visuo-tactile facial stimulation influences sense of identity. AB - Self-face recognition is crucial for sense of identity and self-awareness. Finding self-face recognition disorders mainly in neurological and psychiatric diseases suggests that modifying sense of identity in a simple, rapid way remains a "holy grail" for cognitive neuroscience. By touching the face of subjects who were viewing simultaneous touches on a partner's face, we induced a novel illusion of personal identity that we call "enfacement": The partner's facial features became incorporated into the representation of the participant's own face. Subjects reported that morphed images of themselves and their partner contained more self than other only after synchronous, but not asynchronous, stroking. Therefore, we modified self-face recognition by means of a simple psychophysical manipulation. While accommodating gradual change in one's own face is an important form of representational plasticity that may help maintaining identity over time, the surprisingly rapid changes induced by our procedure suggest that sense of facial identity may be more malleable than previously believed. "Enfacement" correlated positively with the participant's empathic traits and with the physical attractiveness the participants attributed to their partners. Thus, personality variables modulate enfacement, which may represent a marker of the tendency to be social and may be absent in subjects with defective empathy. PMID- 19813139 TI - Decisions to shoot in a weapon identification task: The influence of cultural stereotypes and perceived threat on false positive errors. AB - The decision to shoot a gun engages executive control processes that can be biased by cultural stereotypes and perceived threat. The neural locus of the decision to shoot is likely to be found in the anterior cingulate cortex (ACC), where cognition and affect converge. Male military cadets at Norwich University (N=37) performed a weapon identification task in which they made rapid decisions to shoot when images of guns appeared briefly on a computer screen. Reaction times, error rates, and electroencephalogram (EEG) activity were recorded. Cadets reacted more quickly and accurately when guns were primed by images of Middle Eastern males wearing traditional clothing. However, cadets also made more false positive errors when tools were primed by these images. Error-related negativity (ERN) was measured for each response. Deeper ERNs were found in the medial frontal cortex following false positive responses. Cadets who made fewer errors also produced deeper ERNs, indicating stronger executive control. Pupil size was used to measure autonomic arousal related to perceived threat. Images of Middle Eastern males in traditional clothing produced larger pupil sizes. An image of Osama bin Laden induced the largest pupil size, as would be predicted for the exemplar of Middle East terrorism. Cadets who showed greater increases in pupil size also made more false positive errors. Regression analyses were performed to evaluate predictions based on current models of perceived threat, stereotype activation, and cognitive control. Measures of pupil size (perceived threat) and ERN (cognitive control) explained significant proportions of the variance in false positive errors to Middle-Eastern males in traditional clothing, while measures of reaction time, signal detection response bias, and stimulus discriminability explained most of the remaining variance. PMID- 19813140 TI - Pathophysiological basis of translational stroke research. AB - The high incidence and the devastating consequences of stroke call for efficient therapies but despite extensive experimental evidence of neuroprotective improvements, most clinical treatments have failed. The poor translational success is attributed to the inappropriate selection of clinically irrelevant animal models, the inappropriate focus on clinically irrelevant injury pathways and the inappropriate estimation of the length of therapeutic windows. To substantiate this conclusion, the pathophysiology of experimental stroke is reviewed. Particular emphasis is placed on the importance of collateral pathways, the penumbra concept and the viability thresholds of ischaemia, the haemodynamic and molecular mechanisms of injury evolution and the effect of secondary complications, notably inflammation and brain oedema. The comparison of permanent and transient focal ischaemia, on the one hand, and between mechanical and thrombolytic reperfusion, on the other, reveal basic differences in the mechanisms and dynamics of injury evolution which are of paramount importance for the proper targeting and time window of therapeutic interventions. These differences must be considered for adequate modelling of preclinical stroke studies to avoid unsuccessful translation of experimental data to the clinical setting. PMID- 19813141 TI - Metastatic tumours of the central nervous system - a pathological approach. AB - Metastases are the most common tumours of the central nervous system. Histopathological diagnosis remains the most efficient and specific diagnostic procedure that provides the clinician with quick, specific and cost-effective information necessary for the optimal treatment of the patient. Therefore, the pathologist should be acquainted with the potential opportunities to determine the most precise diagnosis in case of metastatic deposits involving the central nervous system. PMID- 19813142 TI - Inhibition of respiratory processes by overabundance of zinc in neuronal cells. AB - Studies suggest that excessive amounts of free zinc ions can cause neuron death by interfering with the energy production process. The sites of the cell oxidation activity of zinc are the glycolytic enzymes, the Krebs cycle components and the respiratory chain. Further consequences of reduced access to energy are: increased production of reactive forms of oxygen, decrease of the mitochondrial membrane potential and decreased level of ATP. Also, the toxicity of zinc accelerates the supply of extra amounts of this element to the mitochondria, which results in their increased permeability. PMID- 19813143 TI - Expression of RCAS1 protein in microglia/macrophages accompanying brain tumours. An immunofluorescence study. AB - The expression of protein RCAS1 (receptor-binding cancer antigen expressed on SiSo cells), possibly involved in the mechanisms of evasion of immune surveillance by tumours, has been studied in brain astrocytomas grade III and IV and in metastatic carcinomas to the brain by means of double immunofluorescence with antibodies against RCAS1 and respectively anti-GFAP (astroglia) or CD68 or CD74 (macrophages/microglia). Expression of RCAS1 has been reported in many types of carcinomas and in some normal cells, including bone marrow macrophages. Nakabayashi and coworkers recently reported expression of RCAS1 in gliomas. So far no attention has been paid to expression of RCAS1 in non-neoplastic cellular elements of tumours such as macrophages and to the expression of RCAS1 in metastatic carcinomas. We found expression of RCAS1 co-localizing with GFAP+ cells of gliomas and with CD68 and CD74 in large macrophages infiltrating metastatic and primary tumours and sometimes in cells which had morphological characteristics of microglia. Moreover, sometimes strong RCAS1 positivity has been found in metastatic carcinomas. Whether expression of RCAS1 in macrophages accompanying brain tumours is of any importance it is not possible to ascertain at present. However, when elucidating the possible role of RCAS1 in tumour biology, it seems to be necessary to include its presence not only in neoplastic cells. PMID- 19813144 TI - Alpha-synuclein inhibits poly (ADP-ribose) polymerase-1 (PARP-1) activity via NO dependent pathway. AB - alpha-Synuclein (ASN) is a brain-enriched protein that functions as a molecular chaperone and regulator of the synaptic vesicle cycle. However, if ASN is overexpressed and in prefibrillar oligomeric forms it activates free radical formation and has been implicated in neurodegeneration. The nuclear target for the free radical cascade is poly (ADP-ribose) polymerase-1 (PARP-1), a DNA binding enzyme and transcriptional regulator that decides on cell survival or death. Our previous data indicated that soluble oligomeric form of ASN significantly stimulated nitric oxide synthase (NOS) activity and by oxidative stress leads to mitochondria failure and cell death. The aim of this study was to investigate the effect of ASN on PARP-1 protein level and on its activity in the rat brain using radiochemical and immunochemical methods. It was found that ASN (10 microM) had no effect on PARP-1 protein level. However, ASN inhibited this enzyme activity by 35% in rat brain cortex and hippocampus investigated together and in striatum by 18%. An inhibitor of constitutive NOS isoform, N(G)-nitro-L arginine (NNLA, 100 microM), partially prevented ASN-evoked PARP-1 inhibition. The NO pool liberated by ASN could be involved in the decrease of PARP-1 activity. The direct interaction between ASN and PARP-1 protein should be taken into consideration. PMID- 19813145 TI - Ultrastructural evidence of amyloid beta-induced autophagy in PC12 cells. AB - Herein we demonstrate that PC12 cells overexpress human amyloid beta precursor protein bearing double Swedish mutation(AbetaPPsw), showing the phenotype characteristic for Alzheimer's disease (AD). Examination of cells at ultrastructural level revealed the intracellular presence of peptide aggregates. Furthermore, autophagy induction was found to be a hallmark of amyloid beta induced cytotoxicity. Importantly, autophagic vacuoles were co-localized within amyloid beta (Abeta) deposits. This suggests the involvement of autophagy in amyloid beta-elicited cell degeneration. PMID- 19813146 TI - Kufs' disease: diagnostic difficulties in the examination of extracerebral biopsies. AB - Kufs' disease or NCL4 (neuronal ceroid lipofuscinosis type 4) is a rare and poorly characterized, adult-onset form of NCL. The mutation in gene CLN, underlying Kufs' disease, still remains unknown. The diagnosis of this disease is difficult because it is based only on clinical and ultrastructural examinations. We report the case of a 45-year-old woman referred to the Neurological Department with suspicion of Creutzfeldt-Jakob disease (CJD). CJD as well as infectious, autoimmune and some lysosomal diseases were excluded. Since clinical symptoms, i.e. psychotic, auditory and visual hallucinations as well as behavioural disturbances, still suggested metabolic or neurodegenerative disease, a skin and muscle biopsy was performed. On ultrastructural examination the muscle biopsy revealed the subsarcolemmal accumulation of lipofuscin, lipofuscin-like and granular osmiophilic deposits (GRODs). The most unique fingerprint deposits (FP) and curvilinear profiles (CP) for diagnosis of Kufs' disease were located in vascular smooth muscle cells (VSMCs). In these cells lipofuscin-like deposits and GRODs were also visible. The fact that FP and CP were found exclusively in VSMCs jointly with clinical and laboratory data allows us to diagnose Kufs' disease in our patient. PMID- 19813147 TI - Ultrastructural features of astrocytes in the cortex of the hippocampal gyrus and in the neocortex of the temporal lobe in an experimental model of febrile seizures and with the use of topiramate. AB - The objective of the current study was ultrastructural assessment of astroglia in specimens of the hippocampal cortex and neocortex of the temporal lobe in our own experimental model of febrile seizures (FS) in rats, as well as the analysis of the influence of a structurally novel broad spectrum anticonvulsant, topiramate (TPM), upon these cells in the CNS regions studied. The current study was inspired by some interesting literature reports on the in vitro investigation into the biological effects of TPM in primary cultures of rat cortical astrocytes and by the lack of data concerning astroglial morphology in vivo in an experimental model with this antiepileptic. In the FS group, the most pronounced changes in the study cell population referred to protoplasmic astroglia and were observed in approximately 3/4 of these cells. The abnormalities were similarly expressed in the two CNS regions studied, in terms of both quantity and quality. They were characterized by considerable swelling and degenerative changes, both in astrocytic perikarya and their processes. Changes were visible in the elements of the granular endoplasmic reticulum and mitochondria, which had a condensed configuration. In the group receiving topiramate directly after the induction of FS, submicroscopic changes in protoplasmic astrocytes were similarly expressed as in the FS group. However, in the group receiving the drug prior to the induction of FS its protective action was observed on the morphology of approximately 1/3 of the population of the protoplasmic astroglial cells. The remaining protoplasmic astrocytes still showed features of considerable or moderately pronounced injury. The beneficial effect of TPM on the ultrastructure of part of the population of the protoplasmic astroglia in the group in which the drug was applied prior to the induction of FS can be explained, among others, by a protective effect of the blood-brain barrier enhanced by the drug administration, as indicated by our earlier findings. PMID- 19813148 TI - Ganglioglioma associated with alterations of NBN gene. A case report. AB - We report a case of a 13-year-old girl with a tumour of the right fronto-parietal region of the brain. The tumour consisted of two components: a well differentiated astroglial component with Rosenthal fibres and a neoplastic neuronal component. The final histopathology established diagnosis of ganglioglioma WHO grade I. The patient was selected from a group of children with central nervous system (CNS) tumours screened for the most common molecular variants in the NBN gene (exons 5 and 6). Molecular analysis revealed the presence of c.511A>G (p.Ile171Val) substitution on one allele. This is the first patient with ganglioglioma and confirmed mutation in the NBN gene. PMID- 19813149 TI - Pilocytic astrocytoma as a predominant component of a recurrent complex type DNT. AB - Dysembryoplastic neuroepithelial tumour (DNT) is a benign lesion of the cerebral hemispheres usually presenting minimal biological activity after surgical excision. We report an unusual case of a 7-year-old girl with a temporal lobe DNT, which recurred four years after subtotal resection of the tumour. In the recurrent lesion we identified pilocytic astrocytoma (PA) as a predominant component of the tumour. Small pieces of the removed tissues also disclosed remnants of DNT. Clinical presentation of the primary tumour consisted of partial simple seizures, while the recurrent tumour manifested with headache and vomiting. Likewise, the radiological appearance of both tumours was different. We conclude that patients with incompletely removed DNT may suffer local recurrence of that tumour. In rare cases development of a secondary, histologically different neoplasm may also occur. PMID- 19813150 TI - Respiratory syncytial virus prevention outreach project. AB - The objective of the study is to determine whether early identification, outreach, and intervention would increase compliance with palivizumab prophylaxis for respiratory syncytial virus prevention for members of a Texas Medicaid managed care organization. The study cohort was selected from administrative claims data. The observational study period ran from October 1, 2006, to March 31, 2007. Letters, educational materials, and postcards were mailed and outbound telephone calls were placed to qualified members. In addition, letters were sent to primary care providers. Of the 1236 identified members, the 2,238 letters, educational materials, and reminder postcards mailed resulted in 1091 injections to 394 identified members. Only 10% of the 394 members completed their qualified doses as suggested by the guideline. Even with the most intensive outreach, compliance is not highly assured. We must explore other strategies to overcome barriers in this preventive medicine program. PMID- 19813152 TI - Health reform 'house calls': TMA meetings help physicians, patients shape debate. PMID- 19813151 TI - Dream realized: new El Paso medical school first on U.S.-Mexico border. PMID- 19813153 TI - Who's responsible? Online tools help physicians estimate patients' costs. PMID- 19813154 TI - TMA to the rescue: fund-raising helps Ike-ravaged physicians recover. PMID- 19813155 TI - Did you measure the intima-media thickness? PMID- 19813156 TI - Transthoracic sonography in the diagnosis of pulmonary diseases: a systematic approach. PMID- 19813157 TI - Real-time tissue elastography versus FibroScan for noninvasive assessment of liver fibrosis in chronic liver disease. AB - PURPOSE: Transient elastography (FibroScan, [TE]) and serum fibrosis markers such as the FibroTest (FT) are established methods for the noninvasive staging of liver fibrosis. A study using real-time elastography (HI-RTE), which is integrated in a conventional ultrasound system, was recently published with comparable results to transient elastography. The aim of the present study was to validate real-time elastography using the formulas calculated in previous studies and to compare the results to transient elastography and FibroTest for the noninvasive assessment of liver fibrosis. MATERIALS AND METHODS: One hundred and thirty-four patients with chronic liver disease and either histological assessment of liver fibrosis (n = 112) or proven liver cirrhosis (n = 22) were included in the study. All patients received TE, HI-RTE, and biochemical evaluation on the same day as presentation. The calculation of the elasticity score of real-time elastography was performed in accordance with the two previously published studies. RESULTS: The Spearman correlation coefficient between transient elastography, real-time elastography and FibroTest with the histological Chevallier score was statistically significant with 0.78, 0.34, and 0.67, respectively (p < 0.01). The diagnostic accuracy expressed as areas under ROC curves was 0.84, 0.69 and 0.85 for the diagnosis of significant fibrosis (F > or = 2), and 0.97, 0.65, and 0.83 for the diagnosis of cirrhosis, respectively. CONCLUSION: Real-time elastography in its present form cannot replace transient elastography for noninvasive assessment of liver fibrosis. PMID- 19813158 TI - Sonoelastographically guided preoperative localization of suspicious breast microcalcifications detected with mammography. PMID- 19813159 TI - [Therapeutic options in sinunasal adenoid cystic carcinomas--a case report and review]. AB - BACKGROUND: Adenoid cystic carcinomas (ACC) in sinunasal compartments are often not completely resectable. We discuss both, surgical margins and functional results preoperatively and postoperative options for adjuvant therapy. The former opinion of a resistance of ACC towards chemotherapy or irradiation seems to be outdated, even though consensus about therapeutic strategies is still missing. MATERIAL AND METHODS: We discuss therapeutic options and compare data from literature regarding the best adjuvant therapy with the case of a 25-year-old patient with an advanced ACC of the left fossa pterygopalatina. Further we discuss alternative therapeutic options like neutron irradiation, chemotherapy or targeted therapy. Following the advice from literature, we performed an eye saving tumor resection by a left transmaxilloethmoidale sphenoidectomy via combined trans- und extranasal approach. We accepted close surgical margins for the benefit of the abandonment of dismembering measures and performed an adjuvant radiochemotherapy with taxol and carboplatin. RESULTS: With the described therapeutic strategy we reached an optimal local tumor control with unlimited visus and without functional and cosmetic restrictions up to now. Periodic staging did not show any local tumor progress or metastatic spread hitherto. CONCLUSION: Organ preserving surgery and adjuvant radiochemotherapy even in combination with taxol and carboplatin seems to be a sufficient therapeutic option in treating advanced sinunasal ACC, and might not have any prognostic disadvantages to radical surgery. PMID- 19813161 TI - [Swelling of the sternoclavicular joint: rare differential diagnosis]. PMID- 19813162 TI - [Eibecker recommendation of the DGMR to question of law of the wish fulfilling medicine]. PMID- 19813163 TI - [Leukoplakia and erythroplakia--two orale precursor lesions]. AB - The oral leukoplakia/erythroplakia is one of the most common epithelial precursor lesions of the oral squamous cell carcinoma. A clinical distinction is drawn between homogeneous and non-homogeneous as well as proliferative verrucous leukoplakia and erythroplakia. Due to the fact that the oral squamous cell carcinoma has a poor prognosis it is very important to decrease morbidity and mortality by systematic primary, secondary and tertiary prevention of the oral leukoplakia/erythroplakia. On the basis of histological classification systems the risk of malignant transformation can be evaluated, a defined therapy can be induced and the prognosis can be derived. PMID- 19813164 TI - Evaluation of GH-IGF-I axis in adult patients with coeliac disease. AB - The aim of this study was to evaluate GH/IGF-I axis and other pituitary functions in adult patients with coeliac disease. For this purpose, twenty-eight adult coeliac patients [20M, 8F:19-74 years; body mass index (BMI): 18.5-28 kg/m (2)] were recruited. Basal thyroid, adrenal and gonadal function, serum IGF-I and PRL, and routine parameters were evaluated. Dynamic GH secretion was carried out by GHRH plus arginine test. In 20 patients, antipituitary antibodies (APA) were also evaluated. Seven out of 28 patients, independently from disease onset and the gluten-free diet (GFD), showed an impaired GH secretion (25%). All were males, 2 with severe growth hormone deficiency (GHD) and 5 with partial GHD. In patients with GHD, as compared to coeliac patients with normal GH secretion, HOMA (2.1+/ 1.2 vs. 0.9+/-0.4) and QUICKI (0.35+/-0.03 vs. 0.39+/-0.02) levels were significantly higher and lower, respectively, while IGF-I levels were slightly lower (17.7+/-3.7 vs. 24.7+/-6.3, p=NS). APA were negative in all 20 patients studied. In conclusion, a significant number of adult coeliac patients show an impaired GH secretion, this alteration being predominant in males and independent from disease onset and diet regimen. Given the absence of APAs, the cause of this pituitary dysfunction remains unclear even if a previous autoimmune involvement in some cases cannot be excluded. PMID- 19813166 TI - Hepatobiliary fascioliasis: a case with unusual radiological features. AB - We report a case of hepatobiliary fascioliasis presenting with unusual radiological findings that have not been reported previously. Imaging studies revealed hepatic cystic pouches communicating with intrahepatic bile ducts. Snail like, oval shaped and conglomerated echogenic particles with no acoustic shadowing, suggesting F. hepatica, were detected in these cystic pouches. In addition, secondary sclerosing cholangitis developed after fascioliasis. PMID- 19813165 TI - Does information available at admission for delivery improve prediction of vaginal birth after cesarean? AB - We sought to construct a predictive model for vaginal birth after cesarean (VBAC) that combines factors that can be ascertained only as the pregnancy progresses with those known at initiation of prenatal care. Using multivariable modeling, we constructed a predictive model for VBAC that included patient factors known at the initial prenatal visit as well as those that only become evident as the pregnancy progresses to the admission for delivery. We analyzed 9616 women. The regression equation for VBAC success included multiple factors that could not be known at the first prenatal visit. The area under the curve for this model was significantly greater ( P < 0.001) than that of a model that included only factors available at the first prenatal visit. A prediction model for VBAC success, which incorporates factors that can be ascertained only as the pregnancy progresses, adds to the predictive accuracy of a model that uses only factors available at a first prenatal visit. PMID- 19813167 TI - Diffusion-weighted MRI of the kidneys in patients with familial Mediterranean fever: initial experience. AB - PURPOSE: To evaluate the feasibility of diffusion-weighted magnetic resonance imaging (DW-MRI) in the assessment of renal function in patients with familial Mediterranean fever (FMF). MATERIALS AND METHODS: Thirty healthy volunteers who had no history of renal disease, hypertension or vascular disease and 60 patients with FMF were included in the study. Transverse diffusion-weighted multisection echo-planar MRI was performed with the following diffusion gradient b values: 0, 111, 222, 333, 444, 556, 667, 778, 889 and 1000 s/mm(2). The apparent diffusion coefficient (ADC) values, urine protein and serum creatinine levels, and glomerular filtration rates of the healthy volunteers, patients with renal involvement, and patients without were compared by using ANOVA test. ADCs of the kidneys were calculated separately for low (ADC(low); b = 0, 111, 222, 333 s/mm(2)), average (ADC(avg); of all b values), and high (ADC(high); b = 778, 889, 1000 s/mm(2)) b values to enable the differentiation of the relative influence of perfusion fraction and true diffusion. ADC(high) reflects almost only diffusion, whereas ADC(low) is composed of both diffusion and perfusion. RESULTS: There was statistically significant difference between ADC(low) values of the FMF patients with renal involvement and the control group (P < 0.05). Negative correlation was found between the duration of disease and ADC(low) values of the kidneys (r = 0.223, P = 0.087). CONCLUSION: DW-MRI of the kidneys might allow early detection of the renal changes in patients with FMF. This might prevent the progression of disease by giving proper medical treatment. Further studies with larger numbers of FMF patients and more experience on MRI technique are required to help define more conclusively the precise role of DW imaging in detection of renal changes. PMID- 19813168 TI - Percutaneous radiofrequency thermal ablation in the management of lung tumors: presentation of clinical experience on a series of 35 patients. AB - PURPOSE: To present our results in a series of 35 patients with malignant pulmonary lesions, who underwent radiofrequency thermal ablation (RFA) during a period of 18 months. MATERIALS AND METHODS: In our institution, 55 RFA sessions under computed tomography (CT) guidance were performed on 48 pulmonary malignant lesions (23 inoperable primary and 25 metastatic) in 35 patients. RESULTS: Total necrosis was noted in 19 primary (82.6%) and in 19 metastatic lesions (76%). In four primary (17.4%) and in six metastatic lesions (14%), partial necrosis was achieved, and a second RFA session was performed. The 6-month spiral CT follow-up demonstrated recurrence in seven lesions (14.5%) (four primary and three metastatic), which were treated with an additional RFA session. Two of the patients who underwent the procedure died of disseminated disease after one year, accounting for a 1-year survival rate of 94.2%. Mean survival was 14.48 +/- 3.3 months. CONCLUSION: RFA is an effective method for treating unresectable lung carcinoma and lung metastases. PMID- 19813169 TI - A rare case of multiple sclerosis and cerebral hemorrhage associated with osteopetrosis. AB - Osteopetrosis, or Albers-Schonberg disease, is a rare hereditary disease characterized by osteoclast dysfunction and consequent diminished bone resorption and disturbed bone building and remodeling, resulting in abnormally dense and brittle bones. Bone marrow failure, pathologic fractures, and neurologic deficits are common. Osteopetrosis is diagnosed on radiographs. Patients have generalized osteosclerosis, and radiographs may show evidence of fractures. We report a case of cerebral hemorrhage and multiple sclerosis associated with the benign adult form of osteopetrosis. PMID- 19813170 TI - Extraperitoneal pelvic myolipoma. AB - Myolipoma is a very rare adipocytic tumor occurring most frequently in adults, and usually is located in the retroperitoneum or abdomen. It has been described in the retroperitoneum, spinal cord, orbita, breast, round ligament, subcutaneous tissue, pericardium, rectus sheath of the abdominal wall, and abdominal cavity with attachment to the abdominal wall. Most of these tumors are discovered incidentally and are large when discovered. Radiological findings are nonspecific due to the nonlipomatous component of the tumor. We present radiological findings of a large extraperitoneal pelvic myolipoma adjacent to the anterior abdominal wall, detected incidentally in an elderly woman with a presenting complaint of intractable hiccups. PMID- 19813171 TI - Inner ear lipoma. AB - Intracranial lipomas are uncommon congenital malformations that are often asymptomatic. They are very rarely seen in the inner ear. There are a few case reports in the literature related to intravestibular lipoma. It was also defined in the internal auditory canal and the cerebellopontine angle. We present here a case of an inner ear lipoma that diffusely infiltrated the cochlea, vestibule and the semicircular canals. To our knowledge, this is the first reported case in the literature. PMID- 19813172 TI - Intracranial arterialized venous angioma: case report with new insights from functional brain MRI. AB - We present the case of a 58-year-old man who suffered a left thalamic intracerebral hemorrhage. Brain magnetic resonance imaging (MRI) revealed an incidental venous angioma in the left frontal lobe. Further elucidated by cerebral angiography and functional MRI, this venous angioma exhibited arteriovenous shunting. The arterialized venous angioma represents an uncommon, "mixed" intracranial vascular lesion whose natural history remains unknown. PMID- 19813173 TI - Pelvic retroperitoneal angioleiomyoma mimicking a uterine mass. AB - Angioleiomyoma (vascular myoma) is a rare type of leiomyoma originating from smooth muscle cells and containing thick-walled vessels. There are only a few cases of retroperitoneal angioleiomyoma reported in the literature. Herein, we present the ultrasonography, magnetic resonance imaging and histopathological findings of an asymptomatic patient with a pelvic retroperitoneal angioleiomyoma which radiologically mimicked a uterine mass. PMID- 19813174 TI - Utility of multidetector CT in an emergency setting in acute mesenteric ischemia. AB - Prompt diagnosis of acute mesenteric ischemia is a dilemma which is unfortunately generally made at the irreversible phase of intestinal damage. Besides conventional diagnostic imaging modalities contrast enhanced multidetector computed tomography angiography enables fast and detailed evaluation of the mesenteric circulation and abdominal viscera which provides accurate and rapid diagnosis in the emergency room. PMID- 19813175 TI - Fire-eater's pneumonia characterized by pneumatocele formation and spontaneous resolution. AB - Liquid hydrocarbons derived from petroleum are widely used in industry and in households. Aspiration of massive amounts may lead to an acute and fatal form of exogenous lipoid pneumonia (fire-eater's pneumonia). We present a rare case of chemical pneumonitis following accidental lamp oil aspiration characterized by pneumatocele formation and spontaneous resolution. Initial findings on posteroanterior chest radiographs were smoothly circumscribed lower zone masses with and without air-fluid levels. Chest computed tomography also showed multiple thin-walled cavities with and without air-fluid levels. Complete resolution of findings despite initial severe presentation without corticosteroid or antibiotic treatment is noteworthy. PMID- 19813176 TI - Floating aortic thrombus in a patient with non-Hodgkin's Lymphoma. AB - Aortic thrombus is a rare condition unless there is an underlying wall pathology such as atherosclerosis, aneurysm, dissection, or thrombus within the left heart chambers. It causes visceral or peripheral embolisms, and is fatal, if not treated. These characteristics make early diagnosis and therapy essential. We report here the computed tomography findings of a floating thrombus that hanged on to the normal aortic wall with a thin peduncle and caused peripheral embolism in a 58-year-old lymphoma patient who had no evident source of emboli. PMID- 19813177 TI - Lumbar gossypiboma. AB - Gossypiboma is a mass formed by a retained surgical sponge and reactive tissue. The cases with gossypiboma are usually asymptomatic or with nonspecific symptoms, which delay diagnosis for months or years after surgery. We describe imaging findings in a 43-year-old woman with a symptomatic retained surgical sponge in a lumbar laminectomy site. Ultrasonography, computed tomography, magnetic resonance imaging (MRI), and diffusion-weighted MRI were performed. Gossypiboma should be considered in the differential diagnosis of a mass in a patient with a history of prior surgery. Diffusion-weighted MRI may provide important data for differential diagnosis of gossypiboma. With diffusion-weighted MRI, gossypiboma may be distinguished from an abscess by its low signal intensity and increased apparent diffusion coefficient (ADC) as compared to high signal intensity with low ADC in cases of abscess. PMID- 19813178 TI - Simultaneous determination of itraconazole and hydroxyitraconazole in human plasma by liquid chromatography-isotope dilution tandem mass spectrometry method. AB - A rapid and sensitive liquid chromatography-isotope dilution tandem mass spectrometry method was developed and validated for quantification of itraconazole (ITZ) and its active metabolite hydroxyitraconazole (OH-ITZ ) in human plasma. The plasma samples were extracted with tert-butyl methyl ether and two isotope-labeled internal standards (D5-itraconazole and D5 hydroxyitraconazole) were used. The chromatographic separation was performed on a Capcell Pak C(18) MG III (100 x 2 mm, 5 microm, Shiseido). The protonated ions of analytes were detected in positive ionization in multiple reaction monitoring mode. The plasma method has a lower limit of quantification of 1 ng/mL with a linearity range of 1-500 ng/mL for ITZ and OH-ITZ using 100 microL of plasma. The recoveries of the method were found to be 69.47-71.98% for ITZ and 75.68-82.52% for OH-ITZ. The intra- and inter-batch precision was less than 11% for all quality control samples at concentrations of 2.5, 200 and 400 ng/mL. These results indicate that the method was efficient with a short run time (4.5 min) and acceptable accuracy, precision and sensitivity.The validated method was successfully applied to analysis of human plasma samples in pharmacokinetics study. PMID- 19813179 TI - Low internalised restraint predicts criminal recidivism in young female prisoners. AB - BACKGROUND: The Weinberger Adjustment Inventory (WAI) measures social-emotional adjustment along two dimensions: distress and restraint. Four types of adjustment according to this measure have been shown to correlate with criminal recidivism among young male prisoners: reactive (high distress, low restraint), suppressor (high distress, high restraint), non-reactive (low distress, low restraint) and repressor (low distress, high restraint). AIM: To evaluate the predictive potential of the WAI among young female prisoners. METHODS: Women under 30 years old, consecutively admitted to one of three Norwegian prisons, were asked to complete the WAI. Most of those eligible (102, 94%) did so. Re-conviction data were collected from the National Crime Register 38 months (SD = 9.0) after release. RESULTS: The overall re-conviction rate was 38%. Rates differed according to the four WAI types: 53% in the non-reactive, 50% in the reactive, 22% in the suppressor and 11% in the repressor group (p = 0.006). Kaplan-Meier analyses showed that group differences were explained by the WAI restraint dimension (p = 0.008). Differences on the distress dimension did not influence re conviction. Cox regression analysis (adjusting for age at first court conviction and prior offences) found that women with low restraint scores were almost three times as likely to re-offend as women with high restraint scores. CONCLUSION: The WAI appears to be an effective tool for identifying women who are particularly vulnerable to re-offending. Evidence of high capacity for restraint is protective, regardless of distress levels and even after adjusting for the effect of other criminologically important factors. The findings are suggestive that there may be value in individualising 'treatment' or rehabilitation programmes for prisoners. PMID- 19813180 TI - Factors predicting arrest for homeless persons receiving integrated residential treatment for co-occurring disorders. AB - BACKGROUND: Homeless individuals are at increased risk for health and criminal justice problems. AIMS: The aim of this study was to examine risk factors affecting arrest rates in a cohort of homeless people with co-occurring psychiatric and substance-abuse disorders. METHODS: Baseline data were collected from 96 homeless individuals residing in a residential treatment facility for people with co-occurring disorders. Arrest data were obtained for 2 years following treatment intake. Regression analyses were employed to examine interactions between study variables. RESULTS: One third of the sample was arrested during the 2-year follow-up period, principally for drug offences. People referred to treatment directly from the criminal justice system were four times more likely to re-offend than those referred from other sources. Participants' perceived need for mental-health services reduced risk of arrest while their perception of medical needs increased this risk. CONCLUSIONS: The relationship between referral from a criminal justice source and re-arrest after admission to the treatment facility is unsurprising, and consistent with previous literature, but the suggestion of an independently increased risk in the presence of perceived physical health-care needs is worthy of further study. The lower risk of arrest for people who perceive that they have psychological needs is encouraging. PMID- 19813182 TI - In focus: pest management and Darwin. PMID- 19813181 TI - Comparative study of various normal mode analysis techniques based on partial Hessians. AB - Standard normal mode analysis becomes problematic for complex molecular systems, as a result of both the high computational cost and the excessive amount of information when the full Hessian matrix is used. Several partial Hessian methods have been proposed in the literature, yielding approximate normal modes. These methods aim at reducing the computational load and/or calculating only the relevant normal modes of interest in a specific application. Each method has its own (dis)advantages and application field but guidelines for the most suitable choice are lacking. We have investigated several partial Hessian methods, including the Partial Hessian Vibrational Analysis (PHVA), the Mobile Block Hessian (MBH), and the Vibrational Subsystem Analysis (VSA). In this article, we focus on the benefits and drawbacks of these methods, in terms of the reproduction of localized modes, collective modes, and the performance in partially optimized structures. We find that the PHVA is suitable for describing localized modes, that the MBH not only reproduces localized and global modes but also serves as an analysis tool of the spectrum, and that the VSA is mostly useful for the reproduction of the low frequency spectrum. These guidelines are illustrated with the reproduction of the localized amine-stretch, the spectrum of quinine and a bis-cinchona derivative, and the low frequency modes of the LAO binding protein. PMID- 19813183 TI - Sensory ataxic neuropathy dysarthria and ophthalmoparesis (SANDO) in a sibling pair with a homozygous p.A467T POLG mutation. AB - Two siblings who developed fifth-decade-onset, concurrent progressive sensory ataxia, dysarthria, and ophthalmoparesis were found to be homozygous for the p.A467T mutation of the polymerase gamma (POLG) gene. The clinical course in both subjects was progression to severe disability. The enlarging spectrum of sensory ataxic neuropathies associated with mitochondrial DNA (mtDNA) instability and POLG mutations should be recognized and considered in the differential diagnosis of this unusual presentation. PMID- 19813185 TI - Frequency and predictors of nonalcoholic fatty liver disease in myotonic dystrophy. AB - Nonalcoholic fatty liver disease (NAFLD) is a common chronic liver disease that is strongly associated with insulin resistance. Myotonic dystrophy (DM1) is the most common form of adult-onset muscular dystrophy, and there is a high frequency of insulin resistance due to insulin receptor mRNA splicing defects in muscle tissue. The frequency and predictors of NAFLD in this population have not been described. Thirty-six patients with DM1 were prospectively assessed for the presence of NAFLD and insulin resistance. NAFLD was defined by abnormal liver chemistry tests with ultrasound or pathologic evidence of steatosis in the absence of other liver disease. Abnormal liver chemistry tests were found in 44% of DM1 patients (mean ALT 73 +/- 21 U/L, AST 53 +/- 15 U/L), and 87% were attributable to NAFLD. Clinical predictors of NAFLD included increased insulin resistance by the homeostasis model assessment (HOMA) method (9.5 vs. 4.0 U, P = 0.03), elevated fasting insulin (40.4 vs. 16.1 microIU/ml, P = 0.03), abdominal obesity (98.6 vs. 90.8 cm, P = 0.03), elevated triglycerides (195.7 vs. 136.8 mg/dl, P = 0.02), and elevated total cholesterol (213.6 vs. 180.6 mg/dl, P = 0.02). NAFLD is very common and should be considered in the management of DM1. It is strongly associated with markers of insulin resistance and features of the metabolic syndrome. These findings support the role of peripheral insulin resistance in the pathogenesis of NAFLD. PMID- 19813186 TI - Conduction block of peripheral nerve using high-frequency alternating currents delivered through an intrafascicular electrode. AB - Many diseases are characterized by undesired or pathological neural activity. The local delivery of high-frequency currents has been shown to be an effective method for blocking neural conduction in peripheral nerves and may provide a therapy for these conditions. To date, all studies of high-frequency conduction block have utilized extraneural (cuff) electrodes to achieve conduction block. In this study we show that high-frequency conduction block is feasible using intrafascicular electrodes. PMID- 19813187 TI - Prolonged compound muscle action potential duration in critical illness myopathy. AB - Critical illness myopathy (CIM) is a frequent cause of generalized weakness in the intensive care unit. Prolonged compound muscle action potential (CMAP) durations have been described in this patient population, and this study presents further data on CMAP duration in normal controls and patients with CIM. The findings highlight the importance of testing multiple nerve muscle combinations in weak, critically ill patients. Recognition of this pattern, which has not been widely described, can facilitate the diagnosis of CIM. PMID- 19813188 TI - Immune-mediated necrotizing myopathy associated with statins. AB - We report patients from two neuromuscular centers who were evaluated between the years 2000 and 2008 and met the following criteria: (1) proximal muscle weakness occurring during or after treatment with statins; (2) elevated serum creatine kinase (CK); (3) persistence of weakness and elevated CK despite discontinuation of the statin; (4) improvement with immunosuppressive agents; and (5) muscle biopsy showing necrotizing myopathy without significant inflammation. Twenty-five patients fulfilled our inclusion criteria. Twenty-four patients required multiple immunosuppressive agents. Fifteen patients relapsed after being tapered off immunosuppressive therapy. Exposure to statins prior to onset was significantly higher in patients with necrotizing myopathy (82%) as compared to those with dermatomyositis (18%), polymyositis (24%), and inclusion-body myositis (38%) seen in the same time period. The lack of improvement following discontinuation of statins, the need for immunosuppressive therapy, and frequent relapse when treatment was tapered suggest an immune-mediated etiology for this rare, statin associated necrotizing myopathy. PMID- 19813189 TI - Primary AL amyloid polyneuropathy successfully treated with high-dose melphalan followed by autologous stem cell transplantation. AB - We report 2 patients with polyneuropathy associated with amyloid derived from light chains (AL) who were treated successfully with high-dose melphalan followed by autologous peripheral blood stem cell transplantation (HDM/SCT). Neuropathic symptoms improved in conjunction with normalization of serum-free light chains. In addition to amyloid deposits in tissues, an amyloidogenic light chain itself produced by abnormal plasma cells might be harmful to peripheral nerve function, and thus HDM/SCT seems to be a promising therapy for primary AL amyloid polyneuropathy. PMID- 19813190 TI - Skeletal muscle cell MHC I expression: implications for statin-induced myopathy. AB - Statins can induce necrotizing or inflammatory myopathies in some patients. Increased major histocompatibility complex class I (MHC I) expression has been shown in muscle biopsies of statin-induced myopathy. Therefore, we investigated the effect of statins on the expression of MHC I in muscle cells. Using flow cytometry and polymerase chain reaction (PCR), the rhabdomyosarcoma cell line TE671 and primary cultured skeletal muscle cells (SKMC) were investigated for MHC I expression after incubation with different statins and/or interferon-gamma (IFN gamma). TE671 and SKMC express MHC I in the untreated condition. Statins alone reduced the expression of MHC I in SKMC and had no effect on MHC I in TE671 cells. Statins potentiated the MHC I-inducing effect of IFN-gamma in TE671, but not in SKMC, neither at the protein level nor at the mRNA level. The increased muscle MHC I expression in statin-induced myopathy might not be induced directly by statins themselves. PMID- 19813192 TI - Central nervous system involvement in axonal Charcot-Marie-Tooth disease. PMID- 19813191 TI - In vivo assessment of HCN channel current (I(h)) in human motor axons. AB - The "Trond" protocol of nerve excitability tests has been used widely to assess axonal function in peripheral nerve. In this study, the routine Trond protocol was expanded to refine assessment of cAMP-dependent, hyperpolarization-activated current (I(h)) activity. I(h) activity is generated by hyperpolarization activated, cyclic nucleotide-modulated (HCN) channels in response to hyperpolarization. It limits activity-dependent hyperpolarization, contributes to neuronal automaticity, and is implicated in chronic pain states. Published data regarding I(h) activity in motor nerve are scant. We used additional strong, prolonged hyperpolarizing conditioning stimuli in the threshold electrotonus component of the Trond protocol to demonstrate the time-course of activation of I(h) in motor axons. Fifteen healthy volunteers were tested on four occasions during 1 week. I(h) action was revealed in the threshold electrotonus by the limiting and often reversal, after about 100 ms, of the threshold increase caused by strong hyperpolarizing currents. Statistical analysis by repeated-measures analysis of variance enabled confidence limits to be established for variation between subjects and within subjects. The results demonstrate that, of all the excitability parameters, those dependent on I(h) were the most characteristic of an individual, because variance between subjects was more than four times the variance within subjects. This study demonstrates a reliable method for in vivo assessment of I(h,) and also serves to document the normal variability in nerve excitability properties within subjects. PMID- 19813193 TI - Facioscapulohumeral muscular dystrophy: do neurotrophins play a role? AB - Although the molecular defect of facioscapulohumeral muscular dystrophy (FSHD) is well established and involves the contraction of the polymorphic 3.3 kb D4Z4 repeat on the subtelomeric region of chromosome 4q35, the pathologic effects of this deletion remain largely unknown. As a consequence, no specific treatment for FSHD is at present available. Thus, there is the need to explore new areas in an attempt to better characterize pathophysiological alterations in FSHD that might be useful for managing the disease. Neurotrophins (nerve growth factor, brain derived neurotrophic factor, neurotrophin-3, and neurotrophin-4/5) are a class of proteins involved in the development, maintenance, and function of neurons of the peripheral and central nervous systems. In addition, neurotrophins and their RNAs are expressed in muscle, where they have a role in development and regeneration. In this article we put together the experimental evidence that indicates neurotrophins might be involved in the pathophysiology of FSHD and discuss the possible implications of this assumption. PMID- 19813194 TI - Ros-mediated activation of NF-kappaB and Foxo during muscle disuse. AB - We examined reactive oxygen species as upstream activators of nuclear factor kappaB; (NF-kappaB) and forkhead box O (Foxo) in skeletal muscle during disuse atrophy. Catalase, an enzyme that degrades H2O2, was overexpressed in soleus muscles via plasmid injection prior to 7 days of hindlimb immobilization. The increased catalase activity abolished immobilization-induced transactivation of both NF-kappaB and Foxo and attenuated the loss of muscle mass. Thus, H2O2 may be an important initiator of these signaling pathways that lead to muscle atrophy. PMID- 19813195 TI - Sonographic measurements of longitudinal median nerve sliding in patients following nerve repair. AB - Nerve sliding may be restricted following nerve repair. This could result in increased tension across the repair site and lead to poor functional recovery of the nerve. Ultrasound was used to examine longitudinal median nerve sliding in 10 patients who had previously undergone nerve repair surgery following complete division of the median nerve. The median longitudinal movement in the forearm in response to metacarpophalangeal (MCP) joint movements was 2.15 mm on the injured side, compared with 2.54 mm on the uninjured side, a difference that was significant. There was a significant reduction in nerve sliding following repair (median = 8%, range -8% to 54%; P = 0.02), which correlated with time from injury to surgery (rho = 0.87; P = 0.001). These results indicate that ultrasound can be used as an adjunct assessment tool to monitor both morphology and sliding of the nerve through the repair site. It may have future application in the investigation of patients with persisting functional impairment following primary nerve repair. PMID- 19813196 TI - Use of Evans blue dye to compare limb muscles in exercised young and old mdx mice. AB - Evans blue dye (EBD) is used to mark damaged and permeable muscle fibers in mouse models of muscular dystrophy and as an endpoint in therapeutic trials. We counted EBD-positive muscle fibers and extracted EBD from muscles sampled throughout the hindlimbs in young adult and old mdx mice to determine if the natural variability in morphology would allow measurement of a functional improvement in one limb compared to the contralateral limb. Following one bout of rotarod or treadmill exercise that greatly increased serum creatine kinase levels, the number of EBD(+) muscle fibers in 12-19-month-old mdx mice increased 3-fold, EBD in the muscles increased, and, importantly, contralateral pairs of muscles contained similar amounts of EBD. In contrast, the intra- and interlimb amounts of EBD in 2 7-month-old mdx mice were much too variable. A therapeutic effect can more readily be measured in old mdx mice. These results will be useful in the design of therapy protocols using the mdx mouse. PMID- 19813197 TI - Non-lethal neonatal neuromuscular variant of glycogenosis type IV with novel GBE1 mutations. AB - We report a recent case of the severe congenital variant of glycogen storage disease type IV with prolonged survival. The patient was found to be a compound heterozygote for two novel mutations, a missense mutation in exon 5 (p.H188P, c.563A>C) and a severe mutation in intron 5 (c.691+2T>C). We propose that the genotype and the quality of medical care may account for the severe but non lethal phenotype. PMID- 19813198 TI - Added sampling improves reproducibility of multipoint motor unit estimates. AB - Motor unit number estimation (MUNE) has been used to track motor unit attrition. Studies have used the modified multiple-point stimulation (MPS) technique, collecting three surface motor unit action potentials (sMUAPs) from 3 sites to calculate MUNE. Factoring additional sMUAPs should theoretically improve reproducibility, but the optimal number has not been defined. We evaluated the effect of increased sMUAP sampling on test-retest reproducibility of the modified MPS MUNE technique and found that MUNE reproducibility increased with additional sampling. Muscle Nerve, 2010. PMID- 19813199 TI - Syrian hamster infected with Leishmania infantum: a new experimental model for inflammatory myopathies. AB - Idiopathic inflammatory myopathies (IIMs) are inflammatory disorders of unknown origin. On the basis of clinical, histopathological, and immunological features, they can be differentiated into three major and distinct subsets: dermatomyositis; polymyositis; and inclusion-body myositis. Although a few animal models for IIM are currently available, they lack several characteristic aspects of IIMs. The aim of our study was to examine skeletal muscle involvement in an experimental animal model of visceral leishmaniasis, a disseminated infection caused by the protozoan parasite Leishmania infantum, and to compare features of associated inflammation with those of human IIM. Syrian hamsters infected intraperitoneally with amastigotes of L. infantum were killed at 3 or 4 months post-infection, and the skeletal muscles were studied. Focal inflammation was predominantly observed in the endomysium and, to a lesser extent, in perivascular areas. Degenerating muscle fibers were also found, as well as myonecrosis. Immunofluorescence with confocal laser scanning microscopy was used to characterize the phenotype of inflammatory infiltrates and the distribution of MHC class I and II in muscle biopsies. The infiltrating inflammatory cells consisted mainly of T cells, and CD8(+) T cells were found in non-necrotic muscle fibers that expressed MHC class I on the sarcolemma. In addition to T cells, several macrophages were present. The model we are proposing closely resembles polymyositis and may be useful in studying certain aspects of this disease such as the role of T cells in muscle inflammation and myocytotoxicity, while also providing novel therapeutic targets. PMID- 19813200 TI - Effects of exercise and muscle type on BDNF, NT-4/5, and TrKB expression in skeletal muscle. AB - Muscle-derived neurotrophins are thought to contribute to the adaptation of skeletal muscle to exercise, but the effects of brief exercise interventions on BDNF, NT-4/5, and trkB are not understood. RNA was extracted for RT-PCR from soleus and medial gastrocnemius of Sprague-Dawley rats exercised on a treadmill at speeds up to 20 m/min at 5% incline for 5 or 10 days. BDNF expression was elevated in soleus following 5 days (184%, P < 0.001) but not 10 days of exercise. NT-4/5 and trkB were not affected at either time-point. BDNF mRNA was significantly higher in soleus at rest when compared with medial gastrocnemius (193%, P < 0.05). No significant effects of muscle type were detected for NT-4/5 and trkB. Our results indicate differential control of BDNF expression between soleus and medial gastrocnemius following 5 days of exercise. BDNF may be a protein with an uncharacterized contribution to the acute adaptation of skeletal muscle to exercise, whereas NT-4/5 shows no response. PMID- 19813206 TI - Linearity and reliability of the mechanomyographic amplitude versus dynamic torque relationships for the superficial quadriceps femoris muscles. AB - The purpose of this investigation was to examine the linearity and reliability of the mechanomyographic (MMG) amplitude versus dynamic torque relationships for the vastus lateralis (VL), rectus femoris (RF), and vastus medialis (VM) muscles. Nine healthy men and 11 healthy women performed submaximal to maximal, concentric, isokinetic muscle actions of the leg extensors at 30 degrees s(-1) on two occasions. Surface MMG signals were detected from the VL, RF, and VM of the dominant thigh during both trials. The ranges of the coefficients of determination for the MMG amplitude versus dynamic torque relationships were 0.01 0.94 for the VL, 0.01-0.84 for the RF, and 0.19-0.96 for the VM. The intraclass correlation coefficients for the linear MMG amplitude versus torque slope coefficients were 0.823 (VL), 0.792 (RF), and 0.927 (VM). These results indicate that, when analyzed for individual subjects, the MMG amplitude versus dynamic torque relationships demonstrated inconsistent linearity. When using MMG in the clinical setting, dynamic muscle actions of the superficial quadriceps femoris muscles do not appear to be appropriate for assessing changes in muscle function during strength training. PMID- 19813207 TI - Muscle strength and fatigue in patients with generalized myasthenia gravis. AB - Myasthenia gravis (MG) is characterized by fatigue and fluctuating muscle weakness resulting from impaired neuromuscular transmission (NMT). The objective of this study was to quantify, by direct measurement of muscle force, the strength and fatigue of patients with MG. A maximal voluntary isometric contraction protocol of shoulder abductors was used in conjunction with conventional fatigue and disease-severity instruments. Results from patients with (D-MG) and without (ND-MG) decrement on repetitive nerve stimulation (RNS) of the spinal accessory and axillary nerves were compared with healthy controls. Patients with MG reported greater fatigue than controls. Muscle strength was lowest in the D-MG group, followed by the ND-MG group and controls. Normalized shoulder abduction fatigue and recovery values did not differ between the D-MG and ND-MG groups or controls. The RNS decrement appears to relate best to disease severity and muscle weakness but not to objective measures of fatigue in this population. PMID- 19813208 TI - Prenatal diagnosis of idiopathic infantile arterial calcification with hydrops fetalis. AB - Idiopathic infantile arterial calcification (IIAC) is a rare and nearly always fatal disorder. To date, prenatal diagnosis has been reported in fewer than 10 cases. We describe a series of three cases in which the diagnosis of IIAC was made at 23, 25 and 29 weeks' gestation. All three cases presented with a normal anatomy scan at 20 weeks' gestation with an echogenic intracardiac focus. Follow up scans showed generalized hyperechogenicity and calcification of the walls of the large arteries, particularly the aorta and the iliac arteries. All cases developed hydrops fetalis with cardiomegaly and polyhydramnios later in gestation, resulting in intrauterine fetal death in two cases and neonatal death immediately following delivery in the third. This is the largest case series and the earliest gestational age of prenatal diagnosis of IIAC reported to date. When surveying for the disease, serial scans are important, perhaps from 20 weeks' gestation, with close examination of the iliac and aortic arteries. Detection of echogenic intracardiac focus could be an early marker in patients with a family history of the disease. PMID- 19813209 TI - Study on the applicability of frontomaxillary facial angle in the first-trimester trisomy 21 fetuses in Chinese population. AB - OBJECTIVE: To evaluate the measurement of frontomaxillary facial (FMF) angle at 11 weeks to 13 weeks, 6 days in a Chinese population and its applicability in the screening for fetal trisomy 21. METHOD: In a retrospective study, the stored images for the measurement of fetal nuchal translucency (NT) thickness in a Chinese population from August 2003 to March 2007 were reviewed for the comparison of the FMF angle between 22 trisomy 21 fetuses (study group) with 220 randomly selected normal fetuses with satisfactory images (control group). RESULTS: No association between FMF angle and NT, nasal bone length, or fetal heart rate was shown (P > 0.05). But the FMF angle was significantly greater in the trisomy 21 fetuses (89.7 vs 82.8, P < 0.001). The intraobserver and interobserver agreement were assessed in 9.5% (n = 21) normal cases. Mean (SD) differences of two observers were -0.83 (2.69) degrees (P > 0.05). For each observer, mean (SD) differences between the two paired measurements were -0.98 (3.11) degrees and 0.93 (1.80) degrees , respectively (P > 0.05). CONCLUSION: Our results indicated that measurement of the FMF angle in the first trimester can be incorporated into the screening of trisomy 21 in the Chinese population, and its application is not affected by intraobserver and interobserver variability. PMID- 19813210 TI - Proteomic analysis of liver tissue from HBx-transgenic mice at early stages of hepatocarcinogenesis. AB - The hepatitis B virus X-protein (HBx), a multifunctional viral regulator, participates in the viral life cycle and in the development of hepatocellular carcinoma (HCC). We previously reported a high incidence of HCC in transgenic mice expressing HBx. In this study, proteomic analysis was performed to identify proteins that may be involved in hepatocarcinogenesis and/or that could be utilized as early detection biomarkers for HCC. Proteins from the liver tissue of HBx-transgenic mice at early stages of carcinogenesis (dysplasia and hepatocellular adenoma) were separated by 2-DE, and quantitative changes were analyzed. A total of 22 spots displaying significant quantitative changes were identified using LC-MS/MS. In particular, several proteins involved in glucose and fatty acid metabolism, such as mitochondrial 3-ketoacyl-CoA thiolase, intestinal fatty acid-binding protein 2 and cytoplasmic malate dehydrogenase, were differentially expressed, implying that significant metabolic alterations occurred during the early stages of hepatocarcinogenesis. The results of this proteomic analysis provide insights into the mechanism of HBx-mediated hepatocarcinogenesis. Additionally, this study identifies possible therapeutic targets for HCC diagnosis and novel drug development for treatment of the disease. PMID- 19813211 TI - Atomic force microscopy: a powerful molecular toolkit in nanoproteomics. AB - Analysing microbial cell surface proteins is a challenging task in current microbial proteomic research, which has major implications for drug design, vaccine development, and microbial monitoring. In this context, atomic force microscopy (AFM) has recently emerged has a powerful characterization platform, providing valuable insights into the surface proteome of microbial cells. The aim of this article is to show how advanced AFM techniques, that all have in common functionalization of the AFM tip with specific molecules, can be used to answer pertinent questions related to surface-associated proteins, such as what is their spatial arrangement on the cell surface, and what are the forces driving their interaction with the environment? PMID- 19813212 TI - Crystallin distribution patterns in concentric layers from toad eye lenses. AB - Protein distribution patterns across eye lenses from the Asiatic toad Bufo gargarizans were investigated and individual crystallin classes characterised. Special fractionation that follows the growth mode of the lens was used to yield nine fractions corresponding to layers laid down at different chronological (developmental) stages. Proportions of soluble and insoluble crystallins within each fraction were measured by Bradford assay. Water-soluble proteins in all fractions were separated by size-exclusion HPLC and constituents of each class further characterised by electrophoresis, RP-HPLC and MS analysis. In outer lens layers, alpha-crystallin is the most abundant soluble protein but is not found in soluble proteins in the lens centre. Water-soluble beta-crystallins also decrease from their highest level in the outer lens to negligible mounts in the central lens. The proportion of soluble gamma-crystallin increases significantly towards the lens centre where this is the only soluble protein present. Insoluble protein levels increase significantly towards the lens centre. In B. gargarizans lenses, as with other anurans, the predominant water-soluble protein class is gamma crystallin. No taxon-specific crystallins were found. The relationship between the protein distribution patterns and the functional properties of the lens this species is discussed. PMID- 19813214 TI - Fetal sex assignment by first trimester ultrasound: a Tunisian experience. AB - OBJECTIVE: To assess the feasibility and accuracy of fetal sex identification during the first trimester ultrasound exam. METHODS: A prospective study was carried out on 312 fetuses at 11-14 weeks' gestation. The genital region was examined by transabdominal ultrasound. The angle of the genital tubercle to a horizontal line through the lumbosacral skin was measured. Fetal gender was assigned as male if this angle was > 30 degrees and female when it was < 10 degrees . RESULTS: Sex assignment was feasible in 89.7% and accurate in 85.7% of fetuses. Accuracy was similar in males as in females (87.9% vs 83.3%; NS). However, accuracy increased significantly during the gestational age period in male (Chi-square for trend P = 0.03) but not in female (P = 0.41) fetuses. Compared with singletons, presence of multiple fetuses (n = 12) did not influence feasibility or accuracy (89.2% vs 96% and 85.6% vs 86.4% respectively). In male fetuses, there was a significant increase in the angle of the genital tubercle with increasing crown-rump length(CRL) (r = 0.667; p = 0.025), while in females the angle did not significantly change with increasing gestation. CONCLUSION: Sonographic first trimester sex assignment is highly feasible and accurate. PMID- 19813213 TI - The Shigella dysenteriae serotype 1 proteome, profiled in the host intestinal environment, reveals major metabolic modifications and increased expression of invasive proteins. AB - Shigella dysenteriae serotype 1 (SD1) causes the most severe form of epidemic bacillary dysentery. We present the first comprehensive proteome analysis of this pathogen, profiling proteins from bacteria cultured in vitro and bacterial isolates from the large bowel of infected gnotobiotic piglets (in vivo). Overall, 1061 distinct gene products were identified. Differential display analysis revealed that SD1 cells switched to an anaerobic energy metabolism in vivo. High in vivo abundances of amino acid decarboxylases (GadB and AdiA) which enhance pH homeostasis in the cytoplasm and protein disaggregation chaperones (HdeA, HdeB and ClpB) were indicative of a coordinated bacterial survival response to acid stress. Several type III secretion system effectors were increased in abundance in vivo, including OspF, IpaC and IpaD. These proteins are implicated in invasion of colonocytes and subversion of the host immune response in S. flexneri. These observations likely reflect an adaptive response of SD1 to the hostile host environment. Seven proteins, among them the type III secretion system effectors OspC2 and IpaB, were detected as antigens in Western blots using piglet antisera. The outer membrane protein OmpA, the heat shock protein HtpG and OspC2 represent novel SD1 subunit vaccine candidates and drug targets. PMID- 19813215 TI - A versatile 3D culture model facilitates monitoring of astrocytes undergoing reactive gliosis. AB - A major impediment to CNS repair is the glial scar, which forms following damage and is composed mainly of ramified, 'reactive' astrocytes that inhibit neuronal regrowth. The transition of astrocytes into this reactive phenotype (reactive gliosis) is a potential therapeutic target, but glial scar formation has proved difficult to study in monolayer cultures because they induce constitutive astrocyte activation. Here we demonstrate a 3D collagen gel system in which primary rat astrocytes were maintained in a persistently less reactive state than comparable cells in monolayer, resembling their status in the undamaged CNS. Reactivity, proliferation and viability were monitored and quantified using confocal, fluorescence and time-lapse microscopy, 3D image analysis, RT-PCR and ELISA. To assess the potential of this system as a model of reactive gliosis, astrocytes in 3D were activated with TGFbeta1 to a ramified, reactive phenotype (elevated GFAP, Aquaporin 4, CSPG, Vimentin and IL-6 secretion). This provides a versatile system in which astrocytes can be maintained in a resting state, then be triggered to undergo reactive gliosis, enabling real-time monitoring and quantitative analysis throughout and providing a powerful new tool for research into CNS damage and repair. PMID- 19813216 TI - Utilization of human limbal mesenchymal cells as feeder layers for human limbal stem cells cultured on amniotic membrane. AB - Various cell culture techniques for limbal epithelial cells are currently being used for the transplantation of cultured limbal stem cells. In this study, we explored the possibility of using human limbal mesenchymal cells (HLMCs) as feeder layer for the human limbal epithelial cells (HLECs). Single cell suspension of HLECs was seeded onto denuded amniotic membranes with inactivated 3T3 fibroblasts or HLMCs as feeder layer. Expressions of Cytokeratin 3, Np63 and connexin 43 (Cx43) of the cultured epithelial cells were determined at 28 days and the ultrastructure of the epithelium was examined by transmission electron microscope after 14 days and 28 days of cultivation. In both groups, cells were differentiated into multilayer epithelium at 28 days. Basal cells of the cultured epithelium showed a strong nuclear labeling of Np63, but lacked CK3 and Cx43 expression. Transmission electron microscopy examination showed that there were abundant desmosomal contacts between the cells. The key feature the cultured epithelium was occurrence of a typical basement membrane. These results suggested that HLMCs can be used as an alternative feeder layer for HLECs, which makes the bioengineering product biologically safer for the clinical applications. PMID- 19813217 TI - Characteristics of embryo development in Robertsonian translocations' preimplantation genetic diagnosis cycles. AB - OBJECTIVE: To explore the embryo development characteristics in Robertsonian translocations (RTs) in their preimplantation genetic diagnosis (PGD) cycles. METHOD: A total of 37 RT carrier couples underwent 41 blastomere PGD cycles from August 2005 to September 2008. The development of 272 embryos was analyzed in their PGD cycles. RESULT(S): At D3, there were 161 high-grade embryos, including 59 normal/balanced embryos and 102 abnormal embryos. There was no difference between the normal/balanced embryo group and the abnormal embryo group in terms of the high-grade embryo percentage (64.84% vs 56.35%, p = 0.179). However, at D5 D6, the blastocyst percentage in the normal/balanced embryo group was significantly higher than that in the abnormal embryo group (43.96% vs 20.44%, p = 0.000). CONCLUSION(S): Normal/balanced embryos developed better and a self selective mechanism may exist in the RTs' embryos at the blastocyst formation stage. PMID- 19813218 TI - PP13 mRNA expression in the cellular component of maternal blood as a marker for preeclampsia. AB - OBJECTIVE: To assess the PP13 expression in the cellular component of blood in both preeclamptic patients and asymptomatic pregnant women during the early second trimester. METHODS: In the case-control study, peripheral blood samples were obtained from pregnant women with preeclampsia (n = 24) and controls (n = 22). PP13 RNA expression was quantified in the cellular component of the blood by reverse transcription PCR assay. Next, as a cohort study of asymptomatic pregnant women at early gestation, cellular RNA from 41 cases who developed preeclampsia at later gestation and 123 cases of control were analysed, and the possibility of prediction of preeclampsia was assessed. RESULTS: In symptomatic patients, PP13 level in preeclampsia was significantly lower than that in controls (p < 0.001). In the asymptomatic pregnant women, the PP13 level in preeclampsia was significantly lower than that in the controls (p = 0.008). A receiver-operating characteristics (ROC) curve yielded a detection rate of 24 and 31% at a false positive rate of 5 and 10% respectively. CONCLUSION: An alteration in the cellular PP13 RNA expression would affect the placental pathophysiological change. The cellular PP13 expression level could therefore potentially be one of the key markers to predict the clinical onset of preeclampsia. PMID- 19813219 TI - Lysyl oxidase enhances elastin synthesis and matrix formation by vascular smooth muscle cells. AB - Lysyl oxidase (LOX) is a copper-dependent enzyme that initiates covalent crosslinking of elastin precursors by oxidizing peptidyl lysine to aminoadipic semi-aldehydes. Previous studies have shown LOX deficiency to affect crosslinking of elastin and collagen in vivo, resulting in disorganized connective tissue formation. In this study, we investigated the utility of exogenously supplemented LOX peptides (50-100 microl/well) to elastin synthesis, crosslinking efficiency and matrix deposition in adult rat aortic smooth muscle cell (RASMC) cultures. Additionally, we also examined the role of LOX peptides on SMC proliferation and matrix metalloproteinase (MMP) synthesis in these cultures. Highly purified bovine aorta LOX peptide was found to increase matrix elastin synthesis by 40-80% to that in control cultures in a dose-dependent manner, while the crosslinking efficiency significantly (as measured by the ratio of matrix elastin protein to the total elastin protein synthesized) improved to 45-55% of total elastin synthesized under these conditions. However, LOX peptides affected neither SMC proliferation relative to controls, nor elastin precursor (tropoelastin) synthesis, nor the total elastin synthesis on a per-cell basis. In general, LOX peptides also did not affect MMP-2 and MMP-9 activities relative to control cultures, except for MMP-9 activity suppression at a higher LOX dose, suggesting that these LOX peptide cues could be safely used to enhance tropoelastin crosslinking into matrix structures and elastin matrix yield, within tissue engineered constructs, a major challenge in the field. PMID- 19813220 TI - Evaluation of regional left ventricular longitudinal function in 151 normal fetuses using velocity vector imaging. AB - OBJECTIVE: The purpose of this study was to investigate the clinical value of velocity vector imaging (VVI) in the assessment of normal fetal regional myocardial performance and to establish a normative data set for normal Chinese fetuses. METHODS: One hundred and fifty-one healthy Chinese fetuses were divided into five groups according to their gestational age. Digital dynamic four-chamber views were collected and analyzed offline. The regional tissue velocity, strain, and strain rate of the interventricular septum and left lateral wall were measured in systole and diastole. RESULTS: Normal systolic and diastolic values for tissue velocity, strain, and strain rate were established. Tissue velocity decreased gradually from the basal segment to the apical segment (P < 0.01), whereas the strain and strain rate were stable among all segments in every group (P > 0.05). Tissue velocity was dependent on gestational age (P < 0.05), whereas strain and strain rate were stable throughout gestation (P > 0.05). CONCLUSION: VVI is a novel noninvasive tool for quantitative assessment of regional systolic and diastolic function of the fetal heart. Fetal myocardial velocity, strain, and strain rate measurements can be measured easily and reproducibly. This technique is useful for evaluating cardiac function. PMID- 19813221 TI - Application of a first-trimester prediction model for pre-eclampsia based on uterine arteries and maternal history in high-risk pregnancies. AB - OBJECTIVE: To assess the value of a prediction model for pre-eclampsia (PE) in the first trimester (Ultrasound Obstet Gynecol 2007;30:742-794) for the prediction of late (>34 weeks) and early (< or =34 weeks) PE in a high-risk population. METHODS: Longitudinal study performed in 152 high-risk pregnancies with at least one high-risk condition: previous PE, hypertension, pregestational diabetes, renal disease, obesity, hyperlipidemia, autoimmune disorders, thrombophilia or recurrent pregnancy loss. Mean uterine artery pulsatility index at 11 to 13 + 6 weeks and a series of maternal variables were combined in order to obtain the estimated 'a posteriori risk for PE' in each woman. This risk for unaffected women was compared with that for patients who subsequently developed late and early PE. The performance of such approach was described by receiver operating characteristic curves. RESULTS: Late PE developed in 13 (8.6%) pregnancies and early PE in seven (4.6%). The median 'a posteriori risk for PE' in the unaffected, late PE, and early PE groups was 0.62%, 1.22%, and 2.49% (P < 0.01), respectively. For a false-positive rate of 10%, the detection rates of late and early PE were 23.1 and 42.9%, respectively. CONCLUSIONS: This referenced model shows a modest performance when applied to high-risk women. PMID- 19813222 TI - Label-free colorimetric detection of small molecules utilizing DNA oligonucleotides and silver nanoparticles. PMID- 19813223 TI - Acute toxicity of Schizolobium parahyba aqueous extract in mice. AB - The herbal extract of Schizolobium parahyba leaves is used commonly in the Brazil central region to treat snakebites. This study evaluates the acute toxicological effects of Schizolobium parahyba aqueous extract in mice 24 h after intraperitoneal administration. Acute toxicity was evaluated using biochemical, hematological and histopathological assays. Alterations in the levels of transaminases, bilirubin, albumin and prothrombrin time were observed, and these are likely to occur due to hepatic injury, which was confirmed by light microscopy. Liver histopathological analysis revealed the presence of lymph plasmocitary inflammatory infiltrate, but no other histopathological alterations were observed in any of the other organs analysed. The data confirm the low toxicity of the extract of Schizolobium parahyba and provide a model for the selection of a dose that does not cause injuries in the organism. PMID- 19813224 TI - Selective preconcentration of volatile mercaptans in small SPE cartridges: quantitative determination of trace odor-active polyfunctional mercaptans in wine. AB - A general procedure for the selective preconcentration and purification of mercaptans has been developed. Mercaptans are strongly retained in a small (20 mg) SPE cartridge containing p-hydroxymercurybenzoate. The cartridge can then be rinsed with relatively high volumes of polar (water/methanol mixtures) and non polar (pentane or pentane/ether mixtures) rinsing solutions to remove nearly all volatile compounds lacking a thiol functionality. Retained analytes are further eluted with a small volume of an organic solvent containing 1,4-dithioerythritol. Some basic aspects of the strategy, such as the retention of p hydroxymercurybenzoate in the sorbent and its stability versus different rinsing and eluting systems, have been studied in depth. Light sulfur compounds contained in water or wine, including mercaptans such as methanethiol or thioethers, such as diethyl sulfide, can be quantitatively extracted, although only mercaptans can be quantitatively recovered if a polar rinsing is applied. The strategy has been applied to the GC-MS quantitative determination of some trace polyfunctional mercaptans that are key aromas in wine, such as 2-methyl-3-furanthiol, 2 furfurylthiol, 4-mercapto-4-methyl-2-pentanone, 3-mercaptohexyl acetate or 3 mercaptohexanol. The developed method reaches detection limits in the ng/L range and has a satisfactory analytical behavior, being quite simple and fast. PMID- 19813225 TI - Ethyl caffeate from Verdicchio wine: chromatographic purification and in vivo evaluation of its antifibrotic activity. AB - Ethyl caffeate (CfE, caffeic acid ethyl ester) was extracted from dealcoholized Verdicchio, a white wine from Marche (Italy) with ethyl acetate and then purified with semipreparative reversed-phase high-performance liquid chromatography (RP HPLC) using an ODS2 column (25 cm x 20 mm id) at an isocratic flow of 5 mL/min (the mobile phase A was formic acid 4.5% in water and the mobile phase B was acetonitrile). The CfE extract administered intraperitoneally at 1 mumol/L in rats previously treated with 10 mg/kg dimethylnitrosamine was able to prevent the dimethylnitrosamine-induced loss in body and liver weight, as well as to reduce the degree of liver injury, as determined by alanine aminotransferase values and necroinflammatory score, after a 1-week treatment. This was associated with a reduced hepatic stellate cells activation (from 16.8 to 8.3% of smooth muscle actin positive parenchyma) and proliferation (from 11.3 to 5.5 cells/mm(2)). The collagen synthesis was also reduced: the percentage of Sirius Red positive parenchyma decreased from 21.7 to 7.2%. The CfE levels of Verdicchio wine determined with RP-HPLC-DAD were about 14 times the active levels tested in the in vivo test. CfE can be considered as a promising natural compound for future application in chronic liver disease. PMID- 19813226 TI - Qualitative and quantitative differences in the carotenoid composition of yellow and red peppers determined by HPLC-DAD-MS. AB - The carotenoids of a yellow (F(1) Amanda hybrid) and a red (F(1) Magali hybrid) pepper (Capsicum annuum L.) at the ripe stage, marketed in Campinas, Brazil, were identified by the combined use of the chromatographic behavior, absorption spectra, and chemical reactions, confirmed by the mass spectra. HPLC was carried out with a Spherisorb ODS 2 (C18, monomeric, 3 microm, 4.6 x 150 mm) column and DAD and mass detector. The main carotenoids in the yellow pepper were violaxanthin (27.1-36.6 microg/g), lutein (5.3-9.3 microg/g), and beta-carotene (1.6-3.9 microg/g). Neoxanthin, (Z)-violaxanthin, (Z)-antheraxanthin, (Z)-lutein, alpha-cryptoxanthin, beta-cryptoxanthin, zeta-carotene, alpha-carotene, and phytofluene were also found as minor carotenoids. In red pepper, the major carotenoids were capsanthin (29.2-36.2 microg/g), lutein (5.8-8.7 microg/g), beta carotene (5.1-6.8 microg/g), and violaxanthin (1.8-3.6 microg/g). Minor carotenoids were capsorubin, beta-apo-8'-carotenal, antheraxanthin, zeaxanthin, and beta-cryptoxanthin. PMID- 19813227 TI - Determination of metaflumizone residues in cabbage and soil using ultra performance liquid chromatography/ESI-MS/MS. AB - A simple confirmatory method for the determination of metaflumizone in cabbage tissues and soil samples using ultra-performance liquid chromatography-ESI/MS/MS is presented. Metaflumizone residues in cabbage and soils were extracted with acetonitrile and an aliquot was cleaned up with primary and secondary amine. Two precursor product ion transitions for metaflumizone were measured and evaluated to provide the maximum degree of confidence in results. Under ESI negative conditions, quantitation was achieved by monitoring the fragment m/z = 302 and qualitative fragment m/z = 116.5 while also collecting their parent ion m/z = 505.5. Average recoveries for cabbage and soil at four different levels (0.01, 0.05, 1 and 5 mg/kg) ranged from 77.6 to 87.9% with RSD of 3.5-7.9% and RSD(R) of 4.5-7.1%. The coefficients of determination of R(2) > or = 0.9991 were achieved for metaflumizone calibration curves from 0.01 to 5.0 microg/mL. The metaflumizone LODs in cabbage and soil were determined to be both 0.001 mg/kg with a LOQ of 0.004 mg/kg. This method was able to demonstrate quantitative recoveries and provide confirmatory data for the identification of metaflumizone residues in cabbage tissues and the surrounding soils following commercial application. PMID- 19813228 TI - Identification of non-zein proteins in BR473 maize protein bodies by LC-nanoESI MS/MS. AB - The nutritional value of maize seed is limited due to its high content of storage proteins (zeins), which are deficient in essential amino acids such as lysine and tryptophan. In a previous paper, we showed that protein bodies obtained from BR473 maize variety, developed by Embrapa (Brazilian Agricultural Research Corporation), were mainly constituted by Z27 and a smaller quantity of Z50 gamma zeins. Besides zein proteins, other not identified protein band in the SDS/PAGE was also observed, which could indicate the presence of non-zein proteins additionally to gamma-zeins. In the present paper, we have demonstrated the presence of non-zein proteins in BR473 maize protein bodies by LC-nanoESI-MS/MS and database searching. This fact could be related to the excellent energetic value and higher protein quality of BR473 maize grains, since high lysine concentration in some maize varieties has been related to the presence of cytoskeleton proteins that are non-zeins. We have identified the following proteins: Brittle-1 protein (chloroplast precursor), Legumin-1, glyceroldehyde-3 phosphate dehydrogenase, and elongation factor 1-alpha. PMID- 19813229 TI - Study on the migration of bisphenol-A from baby bottles by stir bar sorptive extraction-thermal desorption-capillary GC-MS. AB - Migration of bisphenol-A (BPA), the principal monomer of polycarbonate (PC) baby bottles, was investigated using an aqueous migration simulant. BPA was determined in 200 mL water samples using stir bar sorptive extraction (SBSE) after in situ derivatization with acetic acid anhydride followed by thermal desorption (TD) capillary GC-MS. The concentration of BPA was calculated using the deuterated internal standard d6-BPA. Calibration for BPA was shown to be linear in a concentration range from 1 ng/L to 10 microg/L with a correlation coefficient >0.99. The LOD for BPA (as acetate) was 0.12 ng/L and LOQ 0.40 ng/L (ppt). PC bottles were heated in a water bath and in a microwave oven at four different temperatures (37, 53, 65, and 85 degrees C). The higher the temperature, the more the BPA was released, and after a few heating cycles, the released concentrations became constant. At normal use, i.e. at 37 degrees C, concentrations are ca. 10 ng/L. No significant difference was noted between water bath and microwave heating illustrating that migration of BPA is mainly temperature dependent. PMID- 19813230 TI - Ethanol extract of Fructus Ligustri Lucidi promotes osteogenesis of mesenchymal stem cells. AB - Fructus Ligustri Lucidi (FLL) has been used in traditional Chinese medicine for over 1000 years. The ethanol extract of FLL (EFLL) has been shown to be a potential candidate in the prevention and treatment of osteoporosis. The present study aimed to determine whether EFLL carries out the effect by promoting osteogenesis in mesenchymal stem cells (MSCs). The osteogenic differentiation of MSCs was evaluated by their alkaline phosphatase (ALP) activities and mineralization. Expression of genes was detected by RT-PCR. We found that EFLL significantly stimulated the ALP activities and shortened the time needed for the mineralization of MSCs during osteogenic differentiation. The expression of several osteoblast differentiation regulators was also upregulated by EFLL during this process. Our study demonstrated that the EFLL is capable of enhancing osteogenic differentiation of MSCs. It might be useful for treating diseases with inadequate bone formation, including osteoporosis. PMID- 19813231 TI - Extraction and separation of water-soluble proteins from Bacillus thuringiensis transgenic and non-transgenic maize species by CZE. AB - A new method for extraction and CZE separation and characterization of water soluble proteins (albumin fraction) from Bacillus thuringiensis-transgenic and non-transgenic maize varieties was developed. Protein content in the extracts from maize flours was estimated by UV-absorbance at 280 nm and Lowry protein assay. Protein extracts were analyzed by CZE in four acidic BGEs (100 mM H(3)PO(4), 50 mM Tris, pH 2.25; 500 mM acetic acid, pH 2.6; 200 mM formic acid, 200 mM acetic acid, pH 2.05; 200 mM iminodiacetic acid, pH 2.26) using a homemade CZE device equipped with bare fused silica capillary and UV-absorption detector at 206 nm. Among the tested BGEs, the best resolution of extracted proteins was achieved in isoelectric BGE, 200 mM iminodiacetic acid, pH 2.26. Well-resolved components of protein extracts were characterized by effective electrophoretic mobilities and corrected (migration times normalized) peak areas. Significant qualitative and quantitative differences in CZE-UV profiling were found. This approach can potentially be used to distinguish the B. thuringiensis variety from the wild maize in an easy and straightforward way. PMID- 19813232 TI - Syntheses and biological evaluation of iriomoteolide 3a and analogues. PMID- 19813233 TI - Dehydrocoelenterazine is the organic substance constituting the prosthetic group of Pholasin. PMID- 19813234 TI - Bronsted acid catalyzed Morita-Baylis-Hillman reaction: a new mechanistic view for thioureas revealed by ESI-MS(/MS) monitoring and DFT calculations. AB - A Morita-Baylis-Hillman (MBH) reaction catalyzed by thiourea was monitored by ESI MS(/MS) and key intermediates were intercepted and characterized. These intermediates suggest that thiourea acts as an organocatalyst in all steps of the MBH reaction cycle, including the rate-limiting proton-transfer step. DFT calculations, performed for a model MBH reaction between formaldehyde and acrolein with trimethylamine as base and in the presence or the absence of thiourea, suggest that thiourea accelerates MBH reactions by decreasing the transition-state (TS) energies through bidentate hydrogen bonding throughout the whole catalytic cycle. In the rate-limiting proton-transfer step, the thiourea acts not as a proton shuttle, but as a Bronsted acid stabilizing the basic oxygen center that is formed in the TS. PMID- 19813235 TI - Cisplatin adducts on a GGG sequence within a DNA duplex studied by NMR spectroscopy and molecular dynamics simulations. AB - The antitumor drug cisplatin(cis-[PtCl2(NH3)2]) reacts with cellular DNA to form GG intrastrand adducts between adjacent guanines as predominant lesions. GGG sites have been shown to be hotspots of platination. To study the structural perturbation induced by binding of cisplatin to two adjacent guanines of a GGG trinucleotide,we examined here the decanucleotide duplex d[(G1C2C3G*4 G*5 G6T7 C8G9C10).d(G11C12G13A14C15C16C17G18-G19C20)] (dsCG*G*G) intrastrand cross-linked at the G* guanines by cis-{Pt(NH3)2}2+ using NMR spectroscopy and molecular dynamics (MD) simulations.The NMR spectra of dsCG*G*G were found to be similar to those of previously characterized DNA duplexes cross-linked by cisplatin at apyG*G*X site (py=pyrimidine; X=C,T, A). This similarity of NMR spectra indicates that the base at the 3'-side of the G*G*-Pt cross-link does not affect the structure to a large extent. An unprecedented reversible isomerization between the duplex dsCG*G*G (bearing a G*4 G*5 -Pt chelate) and duplex dsGG*G*T (bearing a G*5 G*6 -Pt chelate)was observed, which yielded a 40:60 equilibrium between the two intrastrand GG-Pt cross-links. No formation of interstrand cross-links was observed.NMR spectroscopic data of dsCG*G*G indicated that the deoxyribose of the 5'-G* adopts an N-type conformation, and the cytidines C3, C15,and C16 have average phase angles intermediate between S and N. The NMR spectroscopic chemical shifts of dsGG*G*T showed some fundamental differences to those of pyG*G* platinum adducts but were in agreement with the NMR spectra reported previously for the DNA duplexes crosslinked at an AG*G*C sequence by cisplatin or oxaliplatin. The presence of apurine instead of a pyrimidine at the 5'-side of the G*G* cross-link seems therefore to affect the structure of the XG* step significantly. PMID- 19813236 TI - Dotz benzannulation reactions: heteroatom and substituent effects in chromium Fischer carbene complexes. AB - We have carried out a theoretical investigation of the Dotz reaction between acetylene and a series of chromium Fischer-type carbenes [(CO)(5)Cr=C(X)R] with different representative substituents (R=CH=CH(2), Ph) and heteroatom ligands (X=OH, NH(2), OCH(3), N(CH(3))(2)) by using density functional theory with the B3LYP functional. We have studied the Dotz and chromahexatriene mechanisms of benzannulation and also the reaction mechanism leading to cyclopentannulation. For the benzannulation, it was found that the most likely mechanism in the case of vinylcarbenes is the chromahexatriene route, whereas for phenylcarbenes, the Dotz route via a ketene intermediate is clearly the most favorable. The reactions leading to the cyclopentannulated and benzannulated products are more exothermic with vinylcarbenes than with phenylcarbenes and also more exothermic with alkoxycarbenes than with aminocarbenes. The relative stability of the cyclopentannulated products as compared with the benzannulated products increases for bulkier X substituents and on going from alkoxy- to aminocarbenes. The kinetic data concurs with the experimental product distribution found for vinylcarbenes, by which mainly benzannulated products are obtained, and dimethylaminophenylcarbenes, which lead exclusively to cyclopentannulated adducts. PMID- 19813237 TI - Multiple reaction pathways in rhodium-catalyzed hydrosilylations of ketones. AB - A detailed density functional theory (DFT) computational study (using the BP86/SV(P) and B3LYP/TZVP//BP86/SV(P) level of theory) of the rhodium-catalyzed hydrosilylation of ketones has shown three mechanistic pathways to be viable. They all involve the generation of a cationic complex [L(n)Rh(I)]+ stabilized by the coordination of two ketone molecules and the subsequent oxidative addition of the silane, which results in the Rh-silyl intermediates [L(n)Rh(III)(H)SiHMe2]+. However, they differ in the following reaction steps: in two of them, insertion of the ketone into the Rh-Si bond occurs, as previously proposed by Ojima et al., or into the Si-H bond, as proposed by Chan et al. for dihydrosilanes. The latter in particular is characterized by a very high activation barrier associated with the insertion of the ketone into the Si-H bond, thereby making a new, third mechanistic pathway that involves the formation of a silylene intermediate more likely. This "silylene mechanism" was found to have the lowest activation barrier for the rate-determining step, the migration of a rhodium-bonded hydride to the ketone that is coordinated to the silylene ligand. This explains the previously reported rate enhancement for R2SiH2 compared to R3SiH as well as the inverse kinetic isotope effect (KIE) observed experimentally for the overall catalytic cycle because deuterium prefers to be located in the stronger bond, that is, C-D versus M-D. PMID- 19813239 TI - Effects of acacia honey on wound healing in various rat models. AB - Honey is a traditional remedy for the treatment of infected wounds, and is becoming more important as microbial resistance to conventional therapeutic agents increases. A study was conducted to assess the wound-healing activity of Acacia honey using incision, excision, burn and dead-space wound models in rats. Different formulations of honey were used and rats were treated topically as well as orally. Both the higher and lower doses of honey produced a significant effect on healing (p < 0.05). The area of epithelization was found to increase, followed by an increase in wound contraction, skin-breaking strength, tissue granulation. The hydroxyproline content also increased in the rats treated with higher doses of honey compared to control, indicating an increase in collagen formation. PMID- 19813238 TI - Structural analysis of glycosphingolipid analogues obtained by the saccharide primer method using CE-ESI-MS. AB - A glycosphingolipid analogue (12-azidododecyl beta-lactoside) as a saccharide primer has been shown to be useful for the synthesis of oligosaccharide libraries by mammalian cells. In the present study, CE-ESI-MS was employed to elucidate the structure of glycosphingolipid analogues derived from 12-azidododecyl beta lactoside (Lac-C12N3) by mammalian cells. MDCK cells and COLO201 cells were cultured with Lac-C12N3, and the glycosylated products secreted into the medium were collected and separated into acidic and neutral products by column chromatography. The acidic products could be directly analyzed by CE-ESI-MS, while the neutral products were converted to anionic derivatives via a reaction with propiolic acid. With this method, it was possible to analyze both acidic and neutral products glycosylated by MDCK cells and COLO201 cells at high sensitivity. PMID- 19813240 TI - Antiobesity effect of baicalin involves the modulations of proadipogenic and antiadipogenic regulators of the adipogenesis pathway. AB - In this study, the antiobesity effects of baicalin, 5,6-dihydroxyflavone-7 glucuronic acid, were characterized using an in vitro system of adipogenesis, i.e. fat cell formation. Baicalin-treatment of 3T3-L1 preadipocytes was shown to inhibit triglyceride accumulation and lipid droplet formation during induced adipogenesis. Microarray analyses showed that baicalin modulated the expression of genes located in pathways such as adipogenesis, cholesterol biosynthesis, focal adhesion and others. In the adipogenesis pathway, treatment with baicalin significantly down-regulated terminal differentiation markers of adipocytes including fatty acid binding protein 4. The effects of baicalin on the core part of the adipogenesis pathway, however, were paradoxical; the expression levels of CCAAT/enhancer binding protein (C/EBP)beta and C/EBPdelta were up-regulated, while the expression levels of the peroxisome proliferator-activated receptor (PPAR)gamma and C/EBPalpha were down-regulated. The antiadipogenic mechanisms of baicalin can be explained by its effects on the upstream part of adipogenesis pathway; baicalin not only up-regulates the antiadipogenic regulators, C/EBPgamma, C/EBP homologous protein and Kruppel-like factor (KLF)2, but also down-regulates the proadipogenic regulator, KLF15. The overall effects of baicalin on these upstream regulators of adipogenesis were antiadipogenic, resulting in the down-regulation of downstream genes and the inhibition of cellular fat accumulation. PMID- 19813242 TI - The relevance of epigenetics to PTSD: implications for the DSM-V. AB - Epigenetic modifications, such as DNA methylation, can occur in response to environmental influences to alter the functional expression of genes in an enduring and potentially, intergenerationally transmissible manner. As such, they may explain interindividual variation, as well as the long-lasting effects of trauma exposure. Although there are currently no findings that suggest epigenetic modifications that are specific to posttraumatic stress disorder (PTSD) or PTSD risk, many recent observations are compatible with epigenetic explanations. These include recent findings of stress-related gene expression, in utero contributions to infant biology, the association of PTSD risk with maternal PTSD, and the relevance of childhood adversity to the development of PTSD. The relevance of epigenetic mechanisms to formulations of PTSD for the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-V) is described. PMID- 19813241 TI - The role of muscle cells in regulating cartilage matrix production. AB - Muscle is one of the tissues located in close proximity to cartilage tissue. Although it has been suggested that muscle could influence skeletal development through generating mechanical forces by means of contraction, very little is known regarding whether muscle cells release biochemical signals to regulate cartilage gene expression. We tested the hypothesis that muscle cells directly regulate cartilage matrix production by analyzing chondrocytes cocultured with muscle cells in 2D or 3D conditions. We found that chondrocytes cultured with C2C12 muscle cells exhibited enhanced alcian blue staining and elevated expression of collagen II and collagen IX proteins. Although nonmuscle cells did not promote cartilage matrix production, converting them into muscle cells enhanced their pro-chondrogenic activity. Furthermore, muscle cell-conditioned medium led to increased cartilage matrix production, suggesting that muscle cells secrete pro-chondrogenic factors. Taken together, our study suggests that muscle cells may play an important role in regulating cartilage gene expression. This result may ultimately lead to the discovery of novel factors that regulate cartilage formation and homeostasis, and provide insights into improving the strategies for regenerating cartilage. PMID- 19813243 TI - Effects of individual control of pH and hypoxia in chondrocyte culture. AB - Effects of oxygen tension (pO(2)) and pH on gene and protein expression and metabolic activity of human chondrocytes were independently assessed. Chondrocytes were cultured under a range of pH (6.4-7.4) and different pO(2) (5 and 20%) during 5 days in a bioreactor. Effects on gene expression, DNA content, protein expression, and metabolic activity were determined. Linear regression analysis showed that gene expression of type I collagen (COL1), SOX9, and VEGF is significantly lower at acidic pH, while expression of aggrecan, type II collagen, and HIF1A is pH-independent. Higher protein levels of VEGF were found under low pO(2). Acidic pH severely lowered VEGF release into medium, glucose consumption, and lactate production. Extracellular pH proved to more potently influence cell function than oxygen tension, the latter showing down-regulation of COL1 gene expression and up-regulation of VEGF protein under hypoxia. Hypoxic culture inhibits COL1 mRNA expression pH-dependently, while expression of SOX9 is largely hypoxia independent, but pH dependent. Expression of HIF1A and VEGF revealed divergent pH dependencies. Subtle fluctuations in extracellular pH and oxygen tension clearly influence chondrocyte metabolism and marker expression. Sophisticated pH and oxygen control not only allows study of (patho)physiological changes, but also opens new venues in cartilage tissue engineering. PMID- 19813244 TI - Convergence of multisensory inputs in Xenopus tadpole tectum. AB - The integration of multisensory information takes place in the optic tectum where visual and auditory/mechanosensory inputs converge and regulate motor outputs. The circuits that integrate multisensory information are poorly understood. In an effort to identify the basic components of a multisensory integrative circuit, we determined the projections of the mechanosensory input from the periphery to the optic tectum and compared their distribution to the retinotectal inputs in Xenopus laevis tadpoles using dye-labeling methods. The peripheral ganglia of the lateral line system project to the ipsilateral hindbrain and the axons representing mechanosensory inputs along the anterior/posterior body axis are mapped along the ventrodorsal axis in the axon tract in the dorsal column of the hindbrain. Hindbrain neurons project axons to the contralateral optic tectum. The neurons from anterior and posterior hindbrain regions project axons to the dorsal and ventral tectum, respectively. While the retinotectal axons project to a superficial lamina in the tectal neuropil, the hindbrain axons project to a deep neuropil layer. Calcium imaging showed that multimodal inputs converge on tectal neurons. The layer-specific projections of the hindbrain and retinal axons suggest a functional segregation of sensory inputs to proximal and distal tectal cell dendrites, respectively. PMID- 19813246 TI - Risk factors disrupting mucosal integrity and subsequent vancomycin-resistant enterococcus infection. PMID- 19813247 TI - Detection of early precursors of t(12;21) positive pediatric acute lymphoblastic leukemia during follow-up. AB - DNA-, RNA-, and cell-based methods provide different biologic information for determining the presence of minimal residual disease (MRD). We monitored the responses of patients with pediatric acute lymphoblastic leukemia (pALL) using DNA markers, TEL/AML1 expression, and scanning fluorescence microscopy (SFM). Using SFM, 36% of patients exhibited 1.5-3.1 log and 2.9-4.2 log higher MRD levels compared with those based on DNA and RNA markers, respectively. CD10+ ancestor cells with germline antigen receptors, but silent TEL/AML1 expression, may reside in the lymphoid stem cell compartment of treated t(12;21)-positive patients and might act as a potential source of cells for late relapses. PMID- 19813245 TI - Captivity reduces hippocampal volume but not survival of new cells in a food storing bird. AB - In many naturalistic studies of the hippocampus wild animals are held in captivity. To test if captivity itself affects hippocampal integrity, adult black capped chickadees (Poecile atricapilla) were caught in the fall, injected with bromodeoxyuridine to mark neurogenesis, and alternately released to the wild or held in captivity. The wild birds were recaptured after 4-6 weeks and perfused simultaneously with their captive counterparts. The hippocampus of captive birds was 23% smaller than wild birds, with no hemispheric differences in volume within groups. Between groups there was no statistically significant difference in the size of the telencephalon, or in the number and density of surviving new cells. Proximate causes of the reduced hippocampal volume could include stress, lack of exercise, diminished social interaction, or limited caching opportunity-a hippocampal-dependent activity. The results suggest the avian hippocampus-a structure essential for rapid, complex relational and spatial learning-is both plastic and sensitive, much as in mammals, including humans. PMID- 19813248 TI - The Quid Pro Quo of pediatric versus adult services for older adolescent cancer patients. AB - PURPOSE: Data from the State of Georgia suggest that pediatric cancers have better survival outcomes when treated at pediatric cancer centers that are members of the nation's Children's Oncology Group (COG). PATIENTS AND METHODS: To determine if the more adult types of cancer that occur in adolescents are better treated at centers with adult oncology expertise, the reported data were re analyzed according to a scale that assessed whether the type of cancer was more likely to have been treated by oncologists with pediatric versus adult cancer experience. RESULTS: The results showed that survival hazard index was linearly correlated in 15- to 19-year-olds with the pediatric versus adult cancer type index (P < 0.0001). All of the five most pediatric type of cancers had a better survival at COG institutions and all of the three tumors with a better survival at non-COG institutions had the highest adult type scores. CONCLUSION: These results demonstrate that adolescent patients with pediatric types of cancer fare better when their care is conducted or supervised by oncologists who specialize in the care of their type of cancer. The Georgia data are among the first to indicate that the more adult type of cancers are better treated on an adult treatment regimen and/or under the supervision or in conjunction with adult treating oncologists. PMID- 19813249 TI - High incidence of hearing loss in long-term survivors of multisystem Langerhans cell histiocytosis. AB - BACKGROUND: Ear involvement in the acute phase of Langerhans cell histiocytosis (LCH) is commonly seen and well documented, but the long-term sequelae are less well described, particularly in relation to hearing loss. METHODS: We investigated 40 patients with biopsy-proven multisystem LCH >5 years from the end of treatment, using detailed audiological assessment and CT/MRI imaging of the petrous temporal bones. RESULTS: The incidence of ear involvement in the acute phase of disease was 70%. Fifteen of the 39 patients tested (38%) had residual permanent hearing loss at long-term follow-up. CONCLUSIONS: The incidence of hearing loss is much higher than has previously been reported in LCH, and may reflect a referral bias of young (<2 years) and more complex patients to our tertiary centre. However, the hearing loss appears to be highly specific to this patient group when compared to other long-term survivors of childhood cancers, probably due to the propensity of LCH to involve the ears. We therefore recommend audiology testing as an important part of long-term follow-up for patients with multisystem LCH. PMID- 19813250 TI - Outcome of childhood acute lymphoblastic leukemia with induction failure treated by the Japan Association of Childhood Leukemia study (JACLS) ALL F-protocol. AB - BACKGROUND: Children with acute lymphoblastic leukemia (ALL) who fail to achieve complete remission (CR) after induction therapy (induction failure: IF) have a poor prognosis; however, there have been few prospective studies in patients with IF. PATIENTS AND METHODS: Between April 1997 and March 2005, 27 of 1,237 leukemic patients (2.2%) failed to achieve CR after four- or five-drug induction therapy. Twenty-three of these patients entered the F-protocol study, which mainly consisted of acute-myeloid-leukemia-oriented chemotherapy followed by scheduled hematopoietic cell transplantation (HCT). RESULTS: Seventeen (73.9%) of the 23 patients responded to re-induction chemotherapy with CR. Of note, 15 (93.8%) of 16 patients with Philadelphia-chromosome-negative (non-Ph(+)) ALL achieved CR; in contrast, only 2 (28.6%) of 7 Ph(+) patients achieved CR. Fourteen (82.4%) of 17 patients remained in CR (CCR) until their scheduled HCT, 12 of the 14 with CCR underwent HCT as scheduled, and 6 patients remain in first CR after a median of 78 months (range, 49-107 months). The 5-year overall survival (OS) rates of 16 patients with non-Ph(+) and 7 patients with Ph(+) were 43.8 +/- 12.4% and 14.3 +/ 13.2%, respectively (P = 0.012). The 5-year OS rate of the 17 patients who obtained CR by re-induction therapy and the 6 who did not were 47.1 +/- 12.1% and 0%, respectively (P < 0.001). CONCLUSION: Acute-myeloid-leukemia-oriented chemotherapy followed by scheduled HCT is a promising treatment strategy for non Ph(+) ALL patients with IF. PMID- 19813251 TI - Acute silent cerebral infarction in children with sickle cell anemia. AB - Silent cerebral infarctions (SCI) occur in up to 35% of children with sickle cell anemia (HbSS) but are rarely recognized during the initial 10-14 days when diffusion weighted magnetic resonance imaging (MRI) can differentiate acute infarctions from remote events. We report acute SCI in seven children with HbSS who had areas of restricted diffusion on MRI without persistent focal neurologic deficits. Four had acute SCI identified following acute anemic events. Our observations suggest that SCI are detectible in the acute phase, present with subtle neurologic symptoms, result in permanent neurologic injury, and may be caused by acute anemic events. PMID- 19813252 TI - Transcranial doppler ultrasonography (TCD) in infants with sickle cell anemia: baseline data from the BABY HUG trial. AB - BACKGROUND: Transcranial Doppler ultrasonography (TCD) is used to predict stroke risk in children with sickle cell anemia (SCA), but has not been adequately studied in children under age 2 years. PROCEDURE: TCD was performed on infants with SCA enrolled in the BABY HUG trial. Subjects were 7-17 months of age (mean 12.6 months). TCD examinations were successfully performed in 94% of subjects (n = 192). RESULTS: No patient had an abnormal TCD as defined in the older child (time averaged maximum mean TAMM velocity > or =200 cm/sec) and only four subjects (2%) had velocities in the conditional range (170-199 cm/sec). TCD velocities were inversely related to hemoglobin (Hb) concentration and directly related to increasing age. CONCLUSION: Determination of whether the TCD values in this very young cohort of infants with SCA can be used to predict stroke risk later in childhood will require analysis of exit TCD's and long-term follow-up, which is ongoing (ClinicalTrials.gov number, NCT00006400). PMID- 19813254 TI - Thymic cyst: a fourth branchial cleft anomaly. AB - We report a unique case of a fourth branchial cleft cyst found within the thymus of an adult patient. In the literature to date, there have been no reports of such a finding in the adult population. These anomalies can often cause recurrent acute suppurative thyroiditis or recurrent deep neck abscesses. Delay in recognizing the underlying etiology can lead to significant complications. PMID- 19813253 TI - Comparison of birth weight corrected for gestational age and birth weight alone in prediction of development of childhood leukemia and central nervous system tumors. AB - INTRODUCTION: High birth weight (HBW) is an established risk factor for childhood acute lymphoblastic leukemia (ALL). The purpose of this study was to evaluate if birth weight (BW) corrected-for-gestational age is a better predictor than BW alone for occurrence of ALL and other malignancies in children. MATERIALS AND METHODS: Birth certificate data of 2,254 children with cancer who were younger than 5 years old at diagnosis and registered at Texas Cancer Registry during 1995 2003 were compared to 11,734 age-matched controls. Multivariable logistic regression was used to compare models with BW corrected-for-gestational age and BW alone. RESULTS: Compared to children who were appropriate for gestational age (AGA), children who were large for gestational age (LGA) at birth had a 1.66 times (95% CI 1.32-2.10) higher odds of ALL. Similarly, children with a BW > or =4,000 g had a 1.5 times (95% CI 1.18-1.89) higher odds for ALL, compared to children who weighed >2,500 and <4,000 g at birth. Using model diagnostics, the model containing BW corrected-for-gestational age was a better predictor than the model with BW alone [Akaike's Information Criterion (AIC) 4,646 vs. 4,658, respectively]. Odds ratios (OR) were similar for LGA children who were <4,000 g and LGA children who were > or =4,000 g (OR = 1.5, 95% CI 0.97-2.5 and OR = 1.67, 95% CI 1.29-2.16, respectively). BW was not an independent risk factor for acute myeloid leukemia or brain tumors. CONCLUSION: BW corrected-for-gestational age is a better predictor than BW alone of risk for ALL. Future studies using BW variable should incorporate gestational age in their analyses. PMID- 19813255 TI - What's in a name? Evidence that Papanicolaou, not Babes, deserves credit for the Pap test. AB - The purpose of our study is to elaborate on the ongoing controversy regarding the origination of the Pap test between the supporters of George Papanicolaou and Aurel Babes. We studied the original articles published by Aurel Babes and George Papanicolaou and conducted a comparative evaluation of both methods. Babes' method is radically different from Papanicolaou's method. Differences included the sampling method, the fixation and staining technique, and the interpretation of the results regarding cases of cervical cancer. We conclude that the establishment of the technique in clinical practice and the idea of its application as preventive control of cervical cancer belong solely to George Papanicolaou. PMID- 19813256 TI - Cytomorphologic characteristics of fine needle core biopsy of multicystic peritoneal mesothelioma: a case report and review of the literature. AB - Multicystic peritoneal mesothelioma (MPM) is an uncommon cystic mesothelial proliferative lesion. It occurs predominantly in women of reproductive age and most commonly arises in the pelvis. The preoperative diagnosis of MPM is difficult to establish based on clinical and radiographic findings, and has therefore traditionally been diagnosed following surgical resection. Due to differing management of MPM and its differential diagnoses including both benign and malignant lesions, it would be beneficial to diagnose MPM preoperatively. We report a case of MPM in a middle aged female that was diagnosed by fine needle core biopsy and touch preparations, allowing for appropriate clinical management. The cytomorphologic features of needle core biopsy, immunocytochemical studies and differential diagnosis are discussed. Furthermore, despite its infrequency, the current case emphasizes the importance of the inclusion of this entity in the differential diagnosis of cystic lesions of the abdomen and pelvis at the time of on-site evaluation and final diagnosis, in order to avoid misinterpretation of strips of benign mesothelial cells as inadequate for diagnosis. PMID- 19813257 TI - Evidence-based evaluation of the risks of malignancy predicted by thyroid fine needle aspiration biopsies. AB - A National Cancer Institute (NCI) "Thyroid Fine-Needle Aspiration (FNA) State of the Science Conference" recently proposed standardized nomenclature and "risks of malignancies" associated with various diagnostic categories. We evaluated the evidence levels of the data used by NCI to predict malignancy risks and whether those estimates had clinical validity in our patient population.Eight hundred seventy-nine patients underwent thyroid FNA during 2006. FNA diagnoses were translated into NCI diagnostic categories, and 2-year follow-up retrospective information was obtained. Four percentages of malignancies were calculated for each diagnostic category using follow-up information from FNA, thyroidectomy, both, and all patients as denominators. 95% confidence intervals (CI) were estimated for all proportions, and results were analyzed with chi-square statistics. "Relative risk" calculations were performed using the percentage of malignancies in the entire population under study as a denominator.Most of the studies cited by the NCI provided incomplete and variable level III evidence based mainly on surgical follow-up. Among our patients, the percentages of malignancies calculated with follow-up data from all patients as the denominator were similar to the "risk estimates" proposed by the NCI, but estimates based on surgical follow-up overestimated the probability of thyroid malignancy for patients with FNA diagnosis of "benign" and "follicular lesions of undetermined significance" (FLUS). Relative risk and 95% CI calculations suggested that the NCI classification could be simplified into three categories: "benign," "FLUS + neoplasm," and "suspicious + malignant." PMID- 19813258 TI - Comparison of the clinical significance of the Papanicolaou test interpretations LSIL cannot rule out HSIL and ASC-H. AB - Despite the two-tiered classification of dysplasia in The Bethesda System (TBS), rare cases fall into the category squamous intraepithelial lesion (SIL) of indeterminate grade. These Pap tests are often interpreted as "LSIL/ASC-H" or "LSIL" with a comment indicating the presence of cells with features approaching HSIL. Patients with LSIL/ASC-H have a significant risk of CIN 2 or worse (29 61.5%) on follow-up cervical biopsies, similar to the risk of CIN 2 or worse in patients with ASC-H Pap tests (24-68%). The purpose of this study was to compare patients with ASC-H and LSIL/ASC-H Pap tests. Women with LSIL/ASC-H had a slightly lower incidence of CIN 2 or worse (PPV = 35.6%, 95% CI: 29.8-41.4%) on follow-up cervical biopsy than the control ASC-H group (PPV = 40.2%, 95% CI: 31.9 56.3%); this difference was not statistically significant. The difference in the distribution of the biopsy results between the two groups was statistically significant (P < 0.001). The current guidelines for the management of cervical cytologic abnormalities from the American Society for Colposcopy and Cervical Pathology (ASCCP) advocate similar treatment algorithms for both LSIL and ASC-H. The main difference is the option of cytologic follow-up or HPV testing for certain "special populations," as an alternative to colposcopy, for LSIL Pap test results. Based on our results, we recommend (1)LSIL/ASC-H to be added to TBS classification and (2) Pap test cases of LSIL/ASC-H may need to be clinically followed in a manner similar to ASC-H, i.e., colposcopy for all patients. PMID- 19813259 TI - Dual fluorescent protein reporters for studying cell behaviors in vivo. AB - Fluorescent proteins (FPs) are useful tools for visualizing live cells and their behaviors. Protein domains that mediate subcellular localization have been fused to FPs to highlight cellular structures. FPs fused with histone H2B incorporate into chromatin allowing visualization of nuclear events. FPs fused to a glycosylphosphatidylinositol anchor signal sequence label the plasma membrane, highlighting cellular shape. Thus, a reporter gene containing both types of FP fusions would allow for effective monitoring of cell shape, movement, mitotic stage, apoptosis, and other cellular activities. Here, we report a binary color coding system using four differently colored FP reporters that generates 16 distinct color codes to label the nuclei and plasma membranes of live cells in culture and in transgenic mice. As an initial test of this system in vivo, the promoter of the human Ubiquitin C (UBC) gene was used to widely express one of the color-code reporters. Widespread expression of the reporter was attained in embryos; however, both male and female transgenic mice were infertile. In contrast, the promoter of the mouse Oct4/Pou5f1 gene linked to two different color-code reporters specifically labeled blastocysts, primordial germ cells, and postnatal germ cells, and these mice were fertile. Time-lapse movies of fluorescently-labeled primordial germs cells demonstrate the utility of the color code system to visualize cell behaviors. This set of new FP reporters should be a useful tool for labeling distinct cell populations and studying their behaviors in complex tissues in vivo. PMID- 19813260 TI - Interstitial deletion of 18q: comparative genomic hybridization array analysis of 46, XX,del(18)(q21.2.q21.33). AB - BACKGROUND: Interstitial deletion of chromosome 18q is rare, making it difficult to assign phenotypes to particular cytogenetic deletions. CASE: We present an 18 year-old female with an interstitial deletion of chromosome 18q21.2-q21.33. The clinical features included severe psychomotor retardation with mild growth retardation, hypotonia, midfacial hypoplasia, carp-shaped mouth, hypertelorism, strabismus, narrow upward slant palpebral fissures, short philtrum, everted lower lip, malformed ears, flat nasal bridge, and epicanthic folds. Brain abnormalities, such as agenesis of the corpus callosum, and abnormalities of the hands and feet were absent. Initially, the deletion was recognized as 18q21.1 q21.31 by conventional chromosomal analysis, and microarray-based comparative genomic hybridization revealed a 9.6-Mb deletion at 18q21.2-q21.33. The deletion included the transcription factor 4 gene and the methyl-CpG binding domain protein 2 (MBD2) gene, but not the MBD1 gene. CONCLUSIONS: The deletion of the transcription factor 4 gene suggested a possible contribution of the deletion to the patient's facial abnormalities, as observed in Pitt-Hopkins syndrome. Together with other reported cases with interstitial deletion of 18q, a possible contribution of haploinsufficiency in both MBD1 and MBD2 genes to a Rett syndrome like phenotype was suggested, but further genetic studies on other cases are necessary to clarify the genotype-phenotype correlation. PMID- 19813261 TI - Gene expression profiling in the fetal cardiac tissue after folate and low-dose trichloroethylene exposure. AB - BACKGROUND: Previous studies show gene expression alterations in rat embryo hearts and cell lines that correspond to the cardio-teratogenic effects of trichloroethylene (TCE) in animal models. One potential mechanism of TCE teratogenicity may be through altered regulation of calcium homeostatic genes with a corresponding inhibition of cardiac function. It has been suggested that TCE may interfere with the folic acid/methylation pathway in liver and kidney and alter gene regulation by epigenetic mechanisms. According to this hypothesis, folate supplementation in the maternal diet should counteract TCE effects on gene expression in the embryonic heart. APPROACH: To identify transcriptional targets altered in the embryonic heart after exposure to TCE, and possible protective effects of folate, we used DNA microarray technology to profile gene expression in embryonic mouse hearts with maternal TCE exposure and dietary changes in maternal folate. RESULTS: Exposure to low doses of TCE (10 ppb) caused extensive alterations in transcripts encoding proteins involved in transport, ion channel, transcription, differentiation, cytoskeleton, cell cycle, and apoptosis. Exogenous folate did not offset the effects of TCE exposure on normal gene expression, and both high and low levels of folate produced additional significant changes in gene expression. CONCLUSIONS: A mechanism by which TCE induces a folate deficiency does not explain altered gene expression patterns in the embryonic mouse heart. The data further suggest that use of folate supplementation, in the presence of this toxin, may be detrimental and not protective of the developing embryo. PMID- 19813262 TI - Synoviocytes are more sensitive than cartilage to the effects of minocycline and doxycycline on IL-1alpha and MMP-13-induced catabolic gene responses. AB - The objective of this study was to determine the primary articular tissue target of doxycycline and minocycline. Synoviocytes-cartilage cocultures (n = 4) were treated with MMP-13 (25 ng/mL medium) or IL-1 (1.0 ng/mL medium) for 24 h. Doxycycline (4.3, 0.43, 0.043 microM) or minocycline (10, 1.0 or 0.1 microM) were then added and cultures were continued for 96 h. Cartilage and media were analyzed for GAG content. Quantitative PCR was used to measure cartilage MMP-3, MMP-13, aggrecan, COL2A1, ADAMTS-4, and ADAMTS-5 expression, and synoviocyte MMP 3, MMP-13, ADAMTS-4, and ADMATS-5 expression. Total and active MMP-3, MMP-13, and ADAMTS 4/5 enzymes were measured in culture medium. All concentrations of doxycycline and minocycline diminished GAG accumulation in the media. All concentrations of minocycline, but only the highest concentration of doxycycline decreased MMP-3 and MMP-13 expression in synoviocytes but not cartilage, and basal ADAMTS-5 mRNA levels in both synoviocytes and cartilage. Only minocycline decreased active MMP-13 protein in synoviocytes. In summary, the protective effects of tetracycline compounds are more pronounced in synoviocytes than cartilage, and following minocycline compared to doxycycline. Studies to determine the molecular mechanism of action of the tetracyclines in synoviocytes might lead to the design of targeted therapeutics for the treatment of OA or RA. PMID- 19813264 TI - Simultaneous determination of flavonoids and phenylethanoids in the flowers of Verbascum densiflorum and V. phlomoides by high-performance liquid chromatography. AB - INTRODUCTION: Mullein (Verbascum) flowers are highly valued herbal drugs used in the treatment of inflammation, asthma, spasmodic coughs and other respiratory tract diseases. Their phenolic constituents are considered to be responsible for the anti-inflammatory and antimicrobial activity of the herb. However, knowledge about the contents of phenolics in flowers is limited and no HPLC method for their analysis is available. OBJECTIVE: To develop and validate an RP-HPLC-UV method for the simultaneous determination of eight flavonoids and two phenylethanoids in the flowers of Verbascum densiflorum and V. phlomoides. METHODOLOGY: HPLC separation was accomplished on a C(18) Lichrosphere 100 column (5 microm, 250 mm x 4.6 mm, i.d.) with an acetonitrile gradient elution using aqueous 0.5% (w/v) orthophosphoric acid solution containing 1% (v/v) tetrahydrofurane. RESULTS: All the calibration curves showed good linear correlation coefficients (r > 0.997) over the wide test ranges. The relative standard deviation of the method was less than 3.4% for intra- and inter-day assays, and the average recoveries were between 93.5 and 101.9%. High sensitivity was demonstrated with detection limits of 0.062-0.083 microg/mL for flavonoid aglycones, 0.156-0.336 microg/mL for flavonoid glycosides and 0.390-0.555 microg/mL for phenylethanoids. The flower samples of V. phlomoides were found to contain high levels of diosmin and tamarixetin 7-rutinoside (2.327-2.392% of dry weight), whereas verbascoside (0.688-0.742% of dry weight) and luteolin 7 glucoside (0.204-0.279% of dry weight) dominated in the V. densiflorum flower. CONCLUSION: The HPLC method established is appropriate for the quality assurance and the differentiation of V. phlomoides and V. densiflorum samples. PMID- 19813263 TI - Comparison of interlaboratory variation in absolute T-cell counts by single platform and optimized dual-platform methods. AB - BACKGROUND: Previous studies have reported that the adoption of a single-platform flow cytometry cell counting method resulted in lower interlaboratory variation in absolute T cell counts as compared to predicate dual-platform flow cytometry methods which incorporate independent automated lymphocyte counts (Schnizlein Bick et al., Clin Diagn Lab Immunol 2000;7:336-343; Reimann et al., Clin Diagn Lab Immunol 2000;7:344-351). In the present study, we asked whether use of a single-platform method could reduce variation in absolute cell counts across the laboratories in the Multicenter AIDS Cohort Study (MACS) (n = 4), as suggested by the studies cited. METHODS: Identical study samples were shipped overnight to the MACS laboratories either by the National Institute of Allergy and Infectious Diseases, Division of AIDS Immunology Quality Assessment (NIAID- IQA) proficiency testing program (n = 14), or by the Los Angeles site of the MACS (n = 10). For each sample, two tubes of blood were received; one was used for an automated complete blood count and differential, and the other for flow cytometry. The latter was performed using both our current dual-platform method (three-color CD45 gating and automated hematology) and the single-platform method (with TruCOUNT beads to generate the absolute counts). RESULTS: The median percent coefficients of variation (%CVs) for the dual-platform and single-platform methods were 6.6 and 9.9, respectively, for CD4 T cell counts, and 5.9 and 8.5, respectively, for CD8 T cell counts (n = 24). These differences were not statistically significant. The differences in absolute T-cell counts between the MACS sites and the median of all laboratories participating in the NIAID-IQA were smaller for the dual-platform than for single-platform absolute count method. CONCLUSION: In contrast to previous reports, we did not observe lower interlaboratory variation across the MACS sites for single-platform absolute lymphocyte subset counting relative to dual-platform methods. This result may be at least partly explained by the lower interlaboratory variation with the optimized dual-platform method in this study relative to the previous reports. PMID- 19813265 TI - Post-fusion treatment with MG132 increases transcription factor expression in somatic cell nuclear transfer embryos in pigs. AB - The objective of this study was to examine the effect of post-fusion treatment of somatic cell nuclear transfer (SCNT) oocytes with the proteasomal inhibitor MG132 on maturation promoting factor (MPF) activity, nuclear remodeling, embryonic development, and gene expression of cloned pig embryos. Immediately after electrofusion, SCNT oocytes were treated with MG132 and/or caffeine for 2 hr, vanadate for 0.5 hr, or vanadate for 0.5 hr followed by MG132 for 1.5 hr. Of the MG132 concentrations tested (0-5 microM), the 1 microM concentration showed a higher rate of blastocyst formation (25.9%) than 0 (14.2%), 0.5 (16.9%), and 5 microM (16.9%). Post-fusion treatment with MG132, caffeine, and both MG132 and caffeine improved blastocyst formation (22.1%, 21.4%, and 24.4%, respectively), whereas vanadate treatment inhibited blastocyst formation (6.5%) compared to the control (11.1%). When examined 2 hr after fusion and 1 hr after activation, MPF activity remained at a higher (P < 0.05) level in SCNT oocytes that were treated post-fusion with caffeine and/or MG132, but it was decreased by vanadate. The rate of oocytes showing premature chromosome condensation was not altered by MG132 but was decreased by vanadate treatment. In addition, formation of single pronuclei was increased by MG132 compared to control and vanadate treatment. MG132-treated embryos showed increased expression of POU5F1, DPPA2, DPPA3, DPPA5, and NDP52l1 genes compared to control embryos. Our results demonstrate that post fusion treatment of SCNT oocytes with MG132 prevents MPF degradation and increases expression of transcription factors in SCNT embryos, which are necessary for normal development of SCNT embryos. PMID- 19813266 TI - Genistein induces receptor and mitochondrial pathways and increases apoptosis during BCL-2 knockdown in human malignant neuroblastoma SK-N-DZ cells. AB - The potent antiapoptotic molecule Bcl-2 is markedly up-regulated in a majority of cancers, including neuroblastoma. Genistein is an isoflavone with antitumor properties. The present study sought to elucidate the molecular mechanism of genistein-induced apoptosis and also to examine the effect of genistein in increasing apoptosis during Bcl-2 knockdown in human malignant neuroblastoma SK-N DZ cells. The cells were transfected with Bcl-2 siRNA plasmid vector, treated with 10 microM genistein, or the combination, and subjected to TUNEL staining and FACS analysis. Semiquantitative and real-time RT-PCR experiments were performed for examining expression of Fas ligand (FasL), tumor necrosis factor-alpha (TNF alpha), Fas-associated death domain (FADD), and TNFR-1-associated death domain (TRADD). The cell lysates were analyzed by Western blotting for levels of molecules involved in both receptor- and mitochondria-mediated apoptotic pathways. Treatment with the combination of Bcl-2 siRNA and genistein resulted in more than 80% inhibition of cell proliferation. TUNEL staining and FACS analysis demonstrated apoptosis in 70% of cells after treatment with the combination of both agents. Apoptosis was associated with increases in Bax:Bcl-2 ratio, mitochondrial release of cytochrome c, and activation of caspases through the mitochondria-mediated apoptotic pathway. Genistein triggered the receptor mediated apoptotic pathway through upregulation of TNF-alpha, FasL, TRADD, and FADD and activation of caspase-8. Combination of Bcl-2 siRNA and genistein triggered a marked increase in cleavage of DFF45 and PARP that resulted in enhanced apoptosis. Our study demonstrates that Bcl-2 knockdown during genistein treatment effectively induced apoptosis in neuroblastoma cells. Therefore, this strategy could serve as a potential therapeutic regimen to inhibit the growth of human malignant neuroblastoma. PMID- 19813267 TI - Effect of mitral regurgitation on cerebrovascular accidents in patients with atrial fibrillation and left atrial thrombus. AB - BACKGROUND: The effect of mitral regurgitation (MR) on the incidence of new cerebrovascular accidents (CVA) and mortality in patients with atrial fibrillation (AF) and left atrial thrombus (LAT) is unknown. OBJECTIVE: To investigate the effect of MR in patients with AF and LAT on new CVA and mortality. METHODS: Eighty nine consecutive patients, mean age 71 years, with AF and LAT documented by transesophageal echocardiography were investigated to determine the prevalence and severity of MR and the association of the severity of MR with new cerebrovascular accidents (CVA) and mortality at 34-mo follow-up. RESULTS: Of 89 patients, 1 + MR was present in 23 patients (26%), 2 + MR in 44 patients (50%), 3 + MR in 17 patients (19%), and 4 + MR in 3 patients (4%). Mean follow-up was 34 +/- 28 mo. The Cox proportional hazards model showed that the severity of increased MR did not significantly increase new CVA or mortality at 34-mo follow-up. The only variable predictive of mortality was left ventricular ejection fraction (LVEF), and with every unit increase in LVEF, the risk decreased by 3%. CONCLUSION: MR occurred in 87 of 89 patients (98%) with AF and LAT. There was no association between the severity of MR and the incidence of CVA or mortality. PMID- 19813268 TI - Comparison of the sensitivity of conventional cytology and the ThinPrep Imaging System for 1,083 biopsy confirmed high-grade squamous lesions. AB - Liquid-based cytology continues to be utilized as an adjunct to conventional cytology in most Australian laboratories, even though a direct-to-vial ThinPrep protocol has been introduced in many countries with established cervical screening programs. Manual screening of ThinPrep slides has been widely practiced for more than 10 years and the recent introduction of the ThinPrep Imaging System (TPI) has been reported as being more sensitive than the conventional smear (CS) in the identification of high-grade cervical disease.We report our experience with ThinPrep Imaging since its introduction into our routine gynecological cytology service. 87,284 split sample pairs reported using the Imaging System demonstrated a decrease in unsatisfactory reports (3.65% for CS and 0.87% for TPI) and an increase in possible high grade and definite high-grade squamous reports (1.57% for CS and 1.62% for TPI).For 1,083 biopsy confirmed high-grade lesions, the correct diagnosis of high grade or possible high-grade squamous disease was made on the ThinPrep imaged slide in 61.0% (661/1,083) of cases and on the CS in 59.4% (643/1,083). This was not statistically significant. When all abnormalities identified on cytology were considered, including possible low grade and definite low-grade abnormalities, the difference in sensitivity for Thinprep imaged slides of 96.0% (1,040/1,083) and CSs of 91.6% (992/1,083) was statistically significant. PMID- 19813269 TI - Anal cytology in patients with AIDS. AB - The objective of the present study was to study the prevalence of abnormal anal cytology in patients with AIDS. Anal smears, obtained with a cytobrush, of 102 HIV-positive patients of the Emilio Ribas Institute (Sao Paulo, Brazil) were collected, and only after that, the patients were submitted to anoscopy. Thirty two patients had LSIL and 14 others had HSIL. Squamous intra-epithelial lesions were also observed in 38% of the patients without condyloma (18/47): in 9 of the 33 patients without history of condyloma (27%) and in 9 of the 14 patients who had previously treated condyloma (64%). An invasive squamous cell carcinoma was observed in one patient without history of condyloma. In all 13 patients with HSIL, biopsies guided by high resolution anoscopy confirmed high grade dysplasia. Our findings suggest that anal cytology is mandatory in AIDS even in patients without macroscopic anal lesions or without previous history of anal condyloma. PMID- 19813270 TI - Epithelioid angiosarcoma: a neoplasm with potential diagnostic challenges. AB - Epithelioid angiosarcomas are extremely rare tumors associated with poor prognosis and early metastases. Its epithelioid cytomorphology and limited vasoformation make it difficult to distinguish from more common malignancies, such as, carcinoma. This can be a potential diagnostic pitfall for the cytopathologist. In this report, the patient is a 24-year-old man presenting with testicular pain, a pelvic mass, and innumerable liver nodules. Immediate interpretation of the needle core biopsies of the pelvic mass and liver lesions initially favored a poorly differentiated adenocarcinoma. Unusual positive immunohistochemical stains for CD30 and CK7 ultimately led the investigation toward a tumor of mesenchymal origin. Further, immunohistochemical evaluation demonstrated positive CD31 and Factor VIII staining and established the final diagnosis of epithelioid angiosarcoma. The tumor cells were negative for CD34, CK20, alpha-fetoprotein, placental-like alkaline phosphatase, hepatocyte paraffin 1, polyclonal carcinoembryonic antigen, CD10, CA-125, prostate-specific antigen, and prostatic acid phosphatase. This case is reported to illustrate the importance of considering the diagnosis of epithelioid angiosarcoma when encountering an "epithelioid" neoplasm particularly with unusual immunoreactivity for CK7 and CD30. PMID- 19813271 TI - Core binding factor at the crossroads: determining the fate of the HSC. AB - Hematopoietic development requires coordinated actions from a variety of transcription factors. The core binding factor (CBF), consisting of a Runx protein and the CBFbeta protein, is a transcription factor complex that is essential for emergence of the hematopoietic stem cell (HSC) from an endothelial cell stage. The hematopoietic defects observed in either Runx1 or CBFbeta knockout mice underscore the necessity of this complex for definitive hematopoiesis. Despite the requirement for CBF in establishing definitive hematopoiesis, Runx1 loss has minimal impact on maintaining the HSC state postnatally, while CBFbeta may continue to be essential. Lineage commitment, on the other hand, is significantly affected upon CBF loss in the adult, indicating a primary role for this complex in modulating differentiation. Given the impact of normal CBF function in the hematopoietic system, the severe consequences of disrupting CBF activity, either through point mutations or generation of fusion genes, are obvious. The physiologic role of CBF in differentiation is subverted to an active process of self-renewal maintenance by the genetic aberrations, through several possible mechanisms, contributing to the development of hematopoietic malignancies including myelodysplastic syndrome and leukemia. The major impact of CBF on the hematopoietic system in both development and disease highlights the need for understanding the intricate functions of this complex and reiterate the necessity of continued efforts to identify potential points of therapeutic intervention for CBF-related diseases. PMID- 19813273 TI - Risk of dying from prostate cancer in men randomized to screening: differences between attendees and nonattendees. AB - BACKGROUND: Although the true benefits and disadvantages of prostate cancer screening are still not known, the analysis of fatal cases is important for increasing knowledge of the effects of prostate cancer screening on mortality. Who dies from prostate cancer despite participation in a population-based prostate-specific antigen (PSA) screening program? METHODS: From the Goteborg branch of the European Randomized study of Screening for Prostate Cancer, 10,000 men randomly assigned to active PSA-screening every second year formed the basis of the present study. Prostate cancer mortality was attributed to whether the men were attendees in the screening program (attending at least once) or nonattendees. RESULTS: Thirty-nine men died from prostate cancer during the first 13 years. Both overall (34% vs 13 %; P<.0001) and cancer-specific mortality (0.8% vs 0.3 %; P<.005) were found to be significantly higher among nonattendees compared with attendees. Furthermore, the majority of deaths (12 of 18) among screening attendees were in men diagnosed at first screening (prevalent cases). Only 6 deaths (including 3 interval cases) were noted among men complying with the biennial screening program. CONCLUSIONS: Nonattendees in prostate cancer screening constitute a high-risk group for both death from prostate cancer and death from other causes comparable to that described in other cancer screening programs. PMID- 19813272 TI - Human papillomavirus vaccination in survivors of childhood cancer. AB - Effective vaccination is now available to prevent human papillomavirus (HPV), the most common sexually transmitted infection and the cause of cervical cancer, which is the second most common cancer among women worldwide. HPV vaccine uptake is particularly important for females surviving cancer, some of whom are at high risk for HPV complications because of the direct and indirect effects of cancer treatment. Thus, version 3.0 of the Children's Oncology Group's Long-Term Follow Up Guidelines for Survivors of Childhood, Adolescent, and Young Adult Cancer recommends HPV vaccination for all eligible females surviving childhood cancer. Because this vaccine was only approved by the US Food and Drug Administration in 2006, little is known regarding the complexity of vaccination uptake among those surviving cancer. The purpose of this article was to describe the HPV vaccine and its usefulness in the survivorship population, provide a rationale for describing cancer survivors as being at increased risk for HPV complications, identify factors associated with HPV vaccination, and discuss the utilization of these predictors in designing strategies to promote adherence to HPV vaccination recommendations within the survivorship context. PMID- 19813274 TI - Outcome and prognostic features in pediatric gliomas: a review of 6212 cases from the Surveillance, Epidemiology, and End Results database. AB - BACKGROUND: Pediatric gliomas are rare and heterogeneous tumors. The Surveillance, Epidemiology, and End Results (SEER) database allows a large-scale analysis of the clinical characteristics and prognostic features of these tumors. METHODS: The authors analyzed available SEER data on 6212 patients younger than 20 years at diagnosis of glioma (1973-2005), according to 4 age categories: <1 year, 1-3 years, 3-5 years, and 5-20 years. RESULTS: The overall 5- and 10-year survival estimates were 71%+/-0.62% (standard error) and 68%+/-0.67%, respectively. Forty-one percent of gliomas were cerebral; the frequency of cerebellar tumors (22%-32% of gliomas) increased sharply after the first year of life. Of the tumors for which grade was available, 77% were low grade (grade I or II). Tumor grade emerged as the most significant independent prognostic factor in all age groups except the youngest age group, in which extent of resection was most significant. Surgery other than gross total resection was an adverse prognostic factor (hazard ratio, 2.18; 95% confidence interval, 1.78-2.67). Age<3 years predicted a greater likelihood of survival in patients with high-grade gliomas and brainstem tumors. Conversely, age<3 years predicted a lower likelihood of survival in patients with low-grade gliomas. Children aged<1 year received less radiotherapy than older patients (P<.0001) and were less likely to undergo gross total resection (P<.0001). CONCLUSIONS: The survival of children with gliomas is influenced by histologic subtype, age, and extent of resection. Despite its limitations, the SEER database provides a useful tool for studies of rare tumors such as pediatric gliomas. PMID- 19813275 TI - Amifostine does not prevent platinum-induced hearing loss associated with the treatment of children with hepatoblastoma: a report of the Intergroup Hepatoblastoma Study P9645 as a part of the Children's Oncology Group. AB - BACKGROUND: The current study was conducted to determine whether amifostine is effective in reducing the toxicities associated with the administration of platinum-containing regimens in children with hepatoblastoma (HB). METHODS: Patients were enrolled on P9645 beginning in March of 1999. Patients who had stage I/II disease received treatment with 4 cycles of combined cisplatin, 5 fluorouracil, and vincristine (C5V) with or without amifostine. Patients who had stage III/IV disease were randomized to receive treatment with 6 cycles of either C5V with or without amifostine or carboplatin alternating with cisplatin (CC) with or without amifostine. Patients who were randomized to receive amifostine were given a dose of 740 mg/m2 intravenously over 15 minutes before each administration of a platinum agent. RESULTS: Eighty-two patients were considered in a special interim analysis of the incidence of toxicity. The disease outcome for patients who received amifostine was similar to the outcome for patients who did not receive amifostine (P=.22). The incidence of significant hearing loss (>40 dB) was similar for patients who did or did not receive amifostine (38% [14 of 37 patients] vs 38% [17 of 45 patients], respectively; P=.68). There were no differences in the incidence of renal or bone marrow toxicities evaluated. Patients who received amifostine had a higher incidence of hypocalcemia (5% vs 0.5%; P=.00006). CONCLUSIONS: Amifostine in the doses and schedule used in this study failed to significantly reduce the incidence of platinum-induced toxicities in patients with HB. PMID- 19813276 TI - Hepatoblastoma, cisplatin, and ototoxicity: good news on deaf ears. PMID- 19813277 TI - Current applications and future prospects of fine-needle aspiration biopsy of locoregional lymph nodes in the management of breast cancer. AB - Staging of disease is routine in the evaluation of patients newly diagnosed with breast cancer. Assessment of palpable and/or nonpalpable locoregional lymph nodes is an important component of the initial staging. Ultrasound (US) is the favored imaging modality for the initial investigation of lymph nodes and results in a significant increase in the specificity of the overall evaluation when used in conjunction with fine-needle aspiration (FNA). This review provides a concise summary, based on published literature, of the current applications and future prospects of FNA biopsy of locoregional lymph nodes in the initial staging and subsequent surgical management of patients with breast cancer. Patients undergo either sentinel lymph node (SLN) biopsy or complete axillary lymph node dissection, based on whether the axillary lymph node status is determined to be negative or positive in the initial staging process. The status of lymph nodes in the supraclavicular, infraclavicular, and internal mammary regions provides more accurate staging information and also impacts subsequent surgical management. The identification and evaluation of intramammary lymph nodes can add value in the overall assessment of patients with breast cancer. The feasibility of noninvasive imaging modalities for SLN mapping in animal models has indicated a good potential for FNA biopsy in the subsequent investigation of SLNs identified noninvasively in humans. PMID- 19813278 TI - Small interfering RNA-mediated down-regulation of SPAG9 inhibits cervical tumor growth. AB - BACKGROUND: The expression of the SPAG9 is associated with various human malignancies. Earlier work revealed a significant association of SPAG9 expression with the early spread of cervical cancer, making it an attractive therapeutic target. Here, the authors investigated the role of SPAG9 in carcinogenesis of squamous cell carcinoma (SCC) of the cervix. Furthermore, they sought to determine whether ablation of SPAG9 expression reduces the tumor growth of cervical SCC in vivo. METHODS: A plasmid-based small interfering RNA approach was used to specifically knock down the expression of SPAG9 in SiHa cells derived from SCC of the cervix in vitro and in vivo. Reverse transcriptase polymerase chain reaction, immunofluorescence staining, flow cytometry, cellular growth, colony formation, migration, invasion, and wound healing assays were studied to characterize SPAG9 in vitro. Furthermore, a cervical cancer xenograft model in nude mice was established to investigate whether knockdown of SPAG9 reduces the tumor growth of cervical SCC in vivo. RESULTS: The results demonstrated that silencing the SPAG9 by small interfering RNA resulted in inhibition of cell growth, colony formation, migration, and invasion. The authors showed for the first time that the knockdown of SPAG9 expression by small interfering RNA significantly suppressed the tumor growth of cervical SCC in vivo. CONCLUSIONS: These results suggest that SPAG9 expression may play a pivotal role in tumor growth and could contribute to the early spread of cervical cancer. Small interfering RNA-mediated down-regulation of SPAG9 represents a promising therapeutic approach for the treatment of cervical cancer. PMID- 19813279 TI - Cysteine-reactive covalent capture tags for enrichment of cysteine-containing peptides. AB - Considering the tremendous complexity and the wide dynamic range of protein samples from biological origin and their proteolytic peptide mixtures, proteomics largely requires simplification strategies. One common approach to reduce sample complexity is to target a particular amino acid in proteins or peptides, such as cysteine (Cys), with chemical tags in order to reduce the analysis to a subset of the whole proteome. The present work describes the synthesis and the use of two new cysteinyl tags, so-called cysteine-reactive covalent capture tags (C3T), for the isolation of Cys-containing peptides. These bifunctional molecules were specifically designed to react with cysteines through iodoacetyl and acryloyl moieties and permit efficient selection of the tagged peptides. To do so, a thioproline was chosen as the isolating group to form, after a deprotection/activation step, a thiazolidine with an aldehyde resin by the covalent capture (CC) method. The applicability of the enrichment strategy was demonstrated on small synthetic peptides as well as on peptides derived from digested proteins. Mass spectrometric (MS) analysis and tandem mass spectrometric (MS/MS) sequencing confirmed the efficient and straightforward selection of the cysteine-containing peptides. The combination of C3T and CC methods provides an effective alternative to reduce sample complexity and access low abundance proteins. PMID- 19813280 TI - Simultaneous delta15N, delta13C and delta34S measurements of low-biomass samples using a technically advanced high sensitivity elemental analyzer connected to an isotope ratio mass spectrometer. AB - Conventional simultaneous CNS stable isotope abundance measurements of solid samples usually require high sample amounts, up to 1 mg carbon, to achieve exact analytical results. This rarely used application is often impaired by high C:S element ratios when organic samples are analyzed and problems such as incomplete conversion into sulphur dioxide occur during analysis. We introduce, as a technical innovation, a high sensitivity elemental analyzer coupled to a conventional isotope ratio mass spectrometer, with which CNS-stable isotope ratios can be determined simultaneously in samples with low carbon content (<40 microg C corresponding to approximately 100 microg dry weight). The system includes downsized reactors, a temperature program-controlled gas chromatography (GC) column and a cryogenic trap to collect small amounts of sulphur dioxide. This modified application allows for highly sensitive measurements in a fully automated operation with standard deviations better than +/-0.47 per thousand for delta15N and delta34S and +/-0.12 per thousand for delta13C (n = 127). Samples collected from one sampling site in a Baltic fjord within a short time period were measured with the new system to get a first impression of triple stable isotope signatures. The results confirm the potential of using delta34S as a stable isotope tracer in combination with delta15N and delta13C measurements to improve discrimination of food sources in aquatic food webs. PMID- 19813281 TI - Sequence confirmation of chemically modified RNAs using exonuclease digestion and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - A broadly applicable, robust, and rapid method for complete sequence confirmation of highly modified oligonucleotides containing a mixture of 2'-deoxy, 2'-fluoro, 2'-o-methyl, abasic and ribonucleotides is presented. The passenger (sense) and guide (antisense) strands from synthetic short interfering RNA duplexes (siRNA) were digested individually using both 5'- and 3'-exonucleases and the resulting ladders were analyzed using matrix-assisted laser desorption/ionization time-of flight (MALDI-TOF) mass spectrometry. Conditions for enzymatic digestion and MALDI-TOF mass analysis were investigated and optimized, and the digestion pattern and sequence coverage of each strand was discussed. Complete sequence confirmation for the antisense strands of four synthetic RNA duplexes was obtained, whereas a three-base sequence gap in the 5'-end was observed for all four sense strands. A general strategy is proposed for routine sequence confirmation of highly modified oligonucleotides, and the potential for complete automation of the method is also discussed. PMID- 19813282 TI - Isotope pattern vector based tandem mass spectral data calibration for improved peptide and protein identification. AB - Tandem mass spectra contain noisy peaks which make peak picking for peptide identification difficult. Moreover, all spectral peaks can be shifted due to systematic measurement errors. In this paper, a novel use of an isotope pattern vector (IPV) is proposed for denoising and systematic measurement error prediction. By matching the experimental IPVs with the theoretical IPVs of candidate fragment ions, true ionic peaks can be identified. Furthermore, these identified experimental IPVs and their corresponding theoretical IPVs are used in an optimization process to predict the systematic measurement error associated with the target spectrum. In return, the subsequent spectral data calibration based on the predicted systematic measurement error enhances the data quality. We show that such an integrated denoising and calibration process leads to significantly improved peptide and protein identification. Different from the commonly employed chemical calibration methods, our IPV-based method is a purely computational method for individual spectra analysis and globally optimizes the use of spectral data. PMID- 19813283 TI - Increasing high-throughput discovery bioanalysis using automated selected reaction monitoring compound optimization, ultra-high-pressure liquid chromatography, and single-step sample preparation workflows. AB - QuickQuan is an integrated software package for Thermo Scientific triple quadrupole mass spectrometers that allows users to automate routine operations ranging from method development to data processing. QuickQuan automated optimization of compound-selected reaction monitoring (SRM) transitions by evaluating both positive and negative polarities during an infusion. Whichever mode produces the most intense Q1 scan is then carried to product ion spectra. QuickQuan then writes these SRM methods to a shared network database. The total volume of compound needed is 100 microL infused over approximately 1.6 min. The auto-optimization is carried out in 96-well plates and does not require an operator present. The SRM database was shared between two identical TSQ Quantum mass spectrometers. For data acquisition, QuickQuan automatically created a sequence file complete with a data processing method pre-populated with compound IDs and corresponding SRM transitions. To increase throughput we coupled each Finnigan Quantum with ultra-high-pressure liquid chromatography (uHPLC) accomplished using 4x Ultra Flux quaternary pumps that were designed to handle pressures up to 15 000 psi. The total run time for all analyses was 1.2 min using BEH 1.7 microm particle C18 columns. Further time reductions were realized with sample preparation accomplished using Strata Impact protein precipitation plates which provided an in-well protein crash and 0.20 micron filtering in a one-step process. Pharmacokinetic data turnaround time was significantly improved by combining these three techniques of automated method development with the speed efficiency of uHPLC and a single step in well sample preparation. PMID- 19813284 TI - Analysis of large historical accurate mass data sets on sector mass spectrometers. PMID- 19813285 TI - Analysis of S-adenosylmethionine and related sulfur metabolites in bacterial isolates of Pseudomonas aeruginosa (BAA-47) by liquid chromatography/electrospray ionization coupled to a hybrid linear quadrupole ion trap and Fourier transform ion cyclotron resonance mass spectrometry. AB - A comprehensive and highly selective method for detecting in bacterial supernatants a modified sulfur nucleoside, S-adenosyl-L-methionine (SAM), and its metabolites, i.e., S-adenosylhomocysteine (SAH), adenosine (Ado), 5'-deoxy-5' methylthioadenosine (MTA), adenine (Ade), S-adenosyl-methioninamine (dcSAM), homocysteine (Hcy) and methionine (Met), was developed. The method is based on reversed-phase liquid chromatography with positive electrospray ionization (ESI+) coupled to a hybrid linear quadrupole ion trap (LTQ) and 7-T Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS). A gradient elution was employed with a binary solvent of 0.05 M ammonium formate at pH 4 and acetonitrile. The assay involves a simultaneous cleanup of cell-free bacterial broths by solid-phase extraction and trace enrichment of metabolites with a 50 fold concentration factor by using immobilized phenylboronic and anion-exchange cartridges. While the quantitative determination of SAM was performed using stable-isotope-labeled SAM-d3 as an internal standard, in the case of Met and Ade, Met-13C and Ade-15N2 were employed as isotope-labeled internal standards, respectively. This method enabled the identification of SAM and its metabolites in cell-free culture of Pseudomonas aeruginosa grown in Davis minimal broth (formulation without sulphur organic compounds), with routine sub-ppm mass accuracies (-0.27 +/- 0.68 ppm). The resulting contents of S(C)S(S)-SAM, S(S) dcSAM, MTA, Ado and Met in the free-cell supernatant of P. aeruginosa was 56.4 +/ 2.1 nM, 32.2 +/- 2.2 nM, 0.91 +/- 0.10 nM, 19.6 +/- 1.2 nM and 1.93 +/- 0.02 microM (mean +/- SD, n = 4 extractions), respectively. We report also the baseline separation (Rs > or = 1.5) of both diastereoisomeric forms of SAM (S(C)S(S) and S(C)R(S)) and dcSAM (S(S) and R(S)), which can be very useful to establish the relationship between the biologically active versus the inactive species, S(C)S(S)/S(C)R(S) and S(S)/R(S) of SAM and dcSAM, respectively. An additional confirmation of SAM-related metabolites was accomplished by a systematic study of their MS/MS spectra. PMID- 19813286 TI - Synthesis and differentiation of alpha- and beta-glycoporphyrin stereoisomers by electrospray tandem mass spectrometry. PMID- 19813287 TI - Cortical activation during finger tracking vs. ankle tracking in healthy subjects. AB - PURPOSE: Explore in healthy subjects whether the differences in cortical activation between ankle and finger movements observed in earlier studies with no attention to accuracy also apply to accuracy-demanding tasks. METHODS: Twenty healthy subjects performed right-finger and right-ankle tracking tasks during functional magnetic resonance imaging. Cortical activation was analyzed in the primary motor area (M1), primary somatosensory area (S1), supplementary motor area (SMA), and premotor cortex (PMC). The blood-oxygen-level-dependent signal intensity of active voxels was compared between the finger and ankle conditions. RESULTS: The results indicated that finger tracking exhibited greater intensity in contralateral (left) M1 and S1 compared to ankle tracking, and ankle tracking exhibited greater intensity in both contralateral and ipsilateral SMA compared to finger tracking. Both M1 and S1 showed more lateralized (contralaterally) organization during finger tracking compared to ankle tracking. Activation maps derived from contrasting the two tracking conditions to each other suggested that ipsilateral areas serve an important role in accuracy-demanding tasks. Regression analysis revealed signal intensity in contralateral M1 and ipsilateral S1 as significant variables in predicting tracking accuracy, but only for finger tracking. CONCLUSIONS: Differences in lateralization between finger and ankle performance found earlier (Kapreli et al., 2006) during simple movement tasks also hold for accuracy-demanding tasks, but dissimilarity in brain organization exists in that the latter show greater ipsilateral activity for both joints. These results invite further research on controlled movements in the lower limb to promote favorable brain reorganization and recovery in people with brain injury. PMID- 19813288 TI - Disparities in mortality patterns among Canadian immigrants and refugees, 1980 1998: results of a national cohort study. AB - This study examines mortality patterns among Canadian immigrants, including both refugees and non-refugees, 1980-1998. Records of a stratified random sample of Canadian immigrants landing between 1980-1990 (N = 369,936) were probabilistically linked to mortality data (1980-1998). Mortality rates among immigrants were compared to those of the general Canadian population, stratifying by age, sex, immigration category, region of birth and time in Canada. Multivariate analysis examined mortality risks for various immigrant subgroups. Although immigrants presented lower all-cause mortality than the general Canadian population (SMR between 0.34 and 0.58), some cause-specific mortality rates were elevated among immigrants, including mortality from stroke, diabetes, infectious diseases (AIDS and hepatitis among certain subgroups), and certain cancers (liver and nasopharynx). Mortality rates differed by region of birth, and were higher among refugees than other immigrants. These results support the need to consider the heterogeneity of immigrant populations and vulnerable subgroups when developing targeted interventions. PMID- 19813289 TI - Health of Chinese illegal immigrants who arrived by boat on the West Coast of Canada in 1999. AB - This was a retrospective review and descriptive analysis of the findings from the medical screening examinations conducted on the illegal migrants from Fujian Province of China (n = 589) who arrived on four boats on the West Coast of Canada between June 14 and September 9, 1999. The Canadian Navy conducted a screening medical exam of the illegal migrants, with Health Canada and Citizenship and Immigration Canada providing suggestions on the format of the exam. The illegal Chinese migrants were predominantly young, male adults. The most prevalent medical conditions detected were dermatological (55.2%), dental problems (25%), trauma (9.2%), urogenital (7.6%), and head/neck (6.6%). Recently induced trauma was more prevalent among females (20.5%) than males (6.5%). One case of community acquired pneumonia was identified and later diagnosed as active pulmonary tuberculosis. Physicians dealing with illegal migrants should look for unusual physical findings and have a higher clinical suspicion regarding infectious diseases (tuberculosis, scabies) and abuse. Future encounters with illegal migrants should include standardized immigration screening exams, with adequate history taking and follow-up. PMID- 19813290 TI - Spousal-abuse among Canadian immigrant women. AB - The study aimed to investigate the rates of self-reported physical and emotional spousal abuse among recent Canadian-immigrant (CI) women compared to Canadian born (CB) women. The study conducted secondary data analyses on the General Social Survey, 1999. A sample of CB (n = 3548) and CI (n = 313) women was drawn that included women 25 to 49 years of age who were currently married or in a common-law relationship. Person weights and bootstrapping estimates were used to estimate the 95% confidence intervals. The proportion of emotional spousal abuse was higher in CI (14.7%, 95% CI: 10.7-18.8%) compared to CB women (8.7%, 95% CI: 7.8-9.6%). However, the proportion of physical spousal abuse was not statistically different between two groups. Possible explanations are discussed setting direction for future research and services for immigrant women. PMID- 19813291 TI - Intersection of Canadian policy parameters affecting women with precarious immigration status: a baseline for understanding barriers to health. AB - Canadian federal policy provides a framework for the immigration and health experiences of immigrant women. The official immigration category under which a migrant is admitted determines to what degree her right to remain in the country (immigration status) is precarious. Women immigrants fall primarily into the more dependent categories and they experience barriers to access to health services arising from this precarious status. Federal immigration and health policies create direct barriers to health through regulation of immigrants' access to services as well as unintended secondary barriers. These direct and secondary policy barriers intersect with each other and with socio-cultural barriers arising from the migrant's socioeconomic and ethno-cultural background to undermine equitable access to health for immigrant women living in Canada. PMID- 19813292 TI - Providing social support for immigrants and refugees in Canada: challenges and directions. AB - In this article we report research findings from a qualitative study of social support for immigrants and refugees in Canada. We focus on challenges from the perspectives of 137 service providers and policymakers in health and immigrant settlement who participated in in-depth interviews and focus groups in three Canadian cities. Results show that social support is perceived to play an important role in immigrant settlement and to have a positive impact on immigrant health, although immigrants face many systemic challenges. Systemic issues- limited resources, lack of integration of policies and programs and narrow service mandates--also limit service providers' abilities to meet newcomer's needs. This research suggests that changes in public discourse about immigrants' contributions, improved governance and service coordination, and a holistic, long term perspective are important to more effectively support immigrant settlement and to promote immigrant health and well being. PMID- 19813293 TI - Pilot survey of HIV risk and contextual problems and issues in Mexican/Latino migrant day laborers. AB - A preliminary survey was conducted with 102 migrant day laborers (MDLs) to assess HIV risk and related contextual problems and issues. These men were primarily Mexican, of low SES background, low in acculturation to the United States, and their income ranged from $100 and $400 a week, 40% of which is sent back home. The psychosocial context of HIV risk included concerns expressed about lack of money and employment, followed by racism, social isolation, sadness and loneliness. High rates of alcohol use and binge drinking that co-occur with sexual activities were reported. While only 7% of MDLs reported illegal injection drug use, needles were frequently shared without bleach cleaning. Men generally did not carry condoms and knowledge of proper condom use was poor. For the most common form of sex reported, vaginal sex, condom use was infrequent. However, men did report confidence in being able to insist on condom use in challenging sexual situations, and they also reported fairly frequent pro-condom attitudes and behaviors within their social circles. Slightly over half of the men reported sexual activity with female partners, during the past 2 months. These female partners were almost evenly divided into regular sex partners, including spouses, and riskier partners such as one time only sex partners, prostitutes, and multiple sex partners. Results also indicated encouraging efforts by MDLs to reduce risk with risky partners (e.g., more condom use). PMID- 19813294 TI - Utilization of preventive care by Haitian immigrants in Miami, Florida. AB - In order to evaluate the health needs and consequences of barriers to health care access for the Haitian immigrant community of Miami-Dade County, Florida, we collected in 2001 demographic and health needs and access data from a probability sample of county residents of Haitian origin. We computed frequencies and prevalence ratios, and employed chi-square and logistic regression methods for data analysis. Hypertension and diabetes were among the most prevalent health conditions mentioned. Up to 39% of participants failed to have an annual physical and 10% failed to receive care for their serious health conditions. On bivariate analysis, insurance coverage, a usual place of care, educational attainment, household income, citizenship status, and duration of residency were associated with services utilization. Citizenship status was the strongest independent predictor of services utilization. The findings suggest that this community faces distinct health needs and daunting challenges to meet those needs. PMID- 19813295 TI - Perceptions of Asian American men about tobacco cigarette consumption: a social learning theory framework. AB - Little information exists regarding the perceptions that ethnic-specific groups of Asian American men have about tobacco cigarette smoking. Thirty Asian American men of immigrant status living in Seattle, Washington, were stratified by ethnicity (Chinese and Vietnamese), language (Mandarin, Cantonese, Vietnamese) and age to comprise six focus groups (two Mandarin speaking men aged 20-40 years and 10 aged 41-65+ years; three Cantonese men aged 20-40 years and another six aged 41-65+ years; four Vietnamese men aged 20-40 years and another five aged 41 65+ years). All group interviews were audio-taped and six separate hard-copy transcripts were produced, independently theme-coded by three investigators to ensure inter-rater reliability, and analyzed with QRS NUD*IST ethnographic software. Bandura (1969, 1986) categorized emergent contextual themes within the constructs of "predisposing, enabling, and reinforcing" behavioral determinants from Social Learning Theory. Smoking to be sociable emerged as the most salient theme. Awareness of tobacco-related diseases other than lung cancer was less evident, as was a self-perceived lack of will-power to quit. Concerns about side stream smoking affecting family members, along with smoking to alleviate stress, were key findings. Further tobacco-related research is needed that incorporates considerations for cultural dynamics. PMID- 19813297 TI - Barriers to health services perceived by Marshallese immigrants. AB - The Marshallese immigrant population, part of a growing Asian American and Pacific Islander population in the United States, has adverse health conditions and disparities that are mainly attributed to their pre-migration health status. Little is known about the perceived and real barriers Marshallese experience in accessing and utilizing health services in the United States. Because of these barriers, their health status is known to exacerbate. This formative study used qualitative methods, using an ethnographic approach, to identify the ethnocultural and socioeconomic barriers to existing health services as perceived by immigrant Marshallese living in Northwest Arkansas. Recommendations were made to improve timely, culturally competent, and appropriate health services. PMID- 19813296 TI - Routine physical examination and forgone health care among Latino adolescent immigrants in the United States. AB - Knowledge concerning patterns of health care utilization among Latino-adolescent immigrants is needed to develop culturally-appropriate programs. The objectives of this study were to estimate the annual prevalence of having had a routine physical exam and episodes of adolescents' not seeking health care when they thought they should (forgone health care) among Latino adolescents by immigrant generational status. Cross-sectional analysis of data from Latino adolescents in Wave I of the National Longitudinal Adolescent Health Study. First-generation immigrants who had lived in the U.S. < or = 5 years were less likely to receive routine care than third-generation immigrants (39.0% vs. 54.9%). This disparity decreased after adjustment for insurance status, parental education and poverty among Mexican origin adolescents. On average, 16.0% of first-generation immigrants who had lived in the U.S. < or = 5 years and 22.5% of third-generation immigrants reported forgoing health care. After adjustment for age, insurance status, parental education and routine care, recent arrivals were less likely than third-generation immigrants to forgo health care. Recent arrivals were less likely to receive a routine physical exam and to forgo care than third-generation immigrants. Future studies should explore the effect of acculturation on knowledge, beliefs and perceptions about health, illness and care-seeking behaviors. PMID- 19813298 TI - Cerebellar infarcts: key features. AB - Cerebellar infarctions often go unrecognized and misdiagnosed. Easily confused with peripheral vestibular system dysfunction, physicians often miss the cardinal symptoms of dizziness and an abnormal gait. If not treated appropriately and quickly, cerebellar infarcts can lead to coma and death. This review discusses the key features of cerebellar infarction, including the anatomical origination and clinical symptomology of the infarcts. Evaluation recommendations include neuroimaging analysis, which can help clarify the etiology and aid in making therapeutic decisions. Management of patients with cerebellar infarcts is similar to that of patients with posterior circulation ischemia. Antithrombotic drugs, thrombolytics, surgery, and angioplasty/stenting are options. PMID- 19813299 TI - The role of autonomic dysfunction in sudden unexplained death in epilepsy patients. AB - The risk of death in people with epilepsy is increased because of disorders that cause epilepsy, known consequences of seizures, and SUDEP (sudden unexplained death in epilepsy). The incidence of sudden death is many times higher in an epilepsy population than for the general public. SUDEP risk increases with the severity of epilepsy but paradoxically affects young adults preferentially. Important risk factors for SUDEP include age 15 to 45 years, refractory epilepsy, tonic-clonic seizures, nocturnal seizures, and periods during which the patient is not observed. Analyses of epidemiologic studies, observations from witnessed near-deaths or deaths, and pathology data have helped focus attention on respiratory and cardiovascular dysfunction as potential mechanisms of SUDEP. Ictal and postictal effects on autonomic functioning and accidental suffocation are commonly cited as potential factors. Monitoring of patients with a history of nocturnal tonic-clonic seizures might help prevent SUDEP. PMID- 19813300 TI - Evaluation of peripheral neuropathy. Part III: vasculitic, infectious, inherited, and idiopathic neuropathies. AB - In this, the third of a 3-part series on peripheral neuropathy, the syndromes of vasculitic, infectious, inherited, and idiopathic neuropathy are discussed. Vasculitis is a frequent cause of neuropathy in the setting of a connective tissue disease. The infectious neuropathies most likely to be encountered in the United States are those due to varicella-zoster virus, human immunodeficiency virus, Lyme disease, hepatitis C virus, and, most recently, West Nile virus. Inherited neuropathies are divided into 2 main types: predominant motor or predominant sensory. The former are generally classed as the Charcot-Marie-Tooth diseases and the latter as the hereditary sensory neuropathies. Each category has a number of different subtypes. If the results of routine screening tests are negative, the clinician must consider special testing for unusual disorders, including evaluations for underlying autoimmune or malignant disorders, genetic tests for inherited neuropathies, and other unusual or selectively ordered tests. These tests are very expensive and should be ordered only after the common causes of neuropathy are excluded. Unless the neuropathy can be substantially alleviated or cured, symptomatic treatment (most often for pain) plays a significant role for these patients. PMID- 19813301 TI - Genotype, proteotype, phenotype relationships in neurodegenerative diseases. Highlights from the 21st Ipsen Foundation Alzheimer's Disease Symposium, September 13, 2004, Paris, France. PMID- 19813302 TI - Multiple sclerosis: causation and putative etiologic factors. Highlights of the 129th Annual Meeting of the American Neurological Association, October 3-6, 2004, Toronto, Ontario, Canada. PMID- 19813303 TI - Report from the 2004 AES Annual Meeting. Highlights from the 58th Annual Meeting of the American Epilepsy Society, December 3-7, 2004, New Orleans, LA. PMID- 19813304 TI - Memory-sparing cognitive deterioration in the elderly: what is the diagnosis? AB - The case presented highlights the difficult differential diagnosis of memory sparing cognitive decline in an elderly patient with previously stable bipolar illness. Many disorders can contribute to cognitive and behavioral deterioration in this population, including reversible causes, particularly delirium and psychiatric illness, and irreversible structural or progressive processes including vascular disease, dementia with Lewy bodies, normal-pressure hydrocephalus, and frontotemporal dementia. PMID- 19813305 TI - A car crash at 10 AM. PMID- 19813306 TI - Sir Albert William Liley. PMID- 19813307 TI - Arbuthnot Lane's cleft-palate needle-holder. PMID- 19813308 TI - John Bell (1763-1820), the 'father' of surgical anatomy. AB - John was one of four very talented sons of the Reverend William Bell. Two qualified as advocates and both became professors. John and his younger brother Charles (later Sir Charles) entered medicine. John qualified with the Edinburgh MD degree in 1779, then obtained the FRCS Edin diploma in 1786. As a student, initially he studied anatomy under Professor Alexander Monro secundus, and it was soon evident to him that his teacher did not have any first-hand knowledge of the problems encountered by surgeons. He then decided to teach anatomy, while practising as a surgeon, and was among the first to emphasize the relevance of anatomy to surgical practice. During this period he lectured and wrote on anatomy and surgical anatomy. After 1800 he, like many of his surgical colleagues, was excluded from the Edinburgh Royal Infirmary as a consequence of the activities of Professor James Gregory, one of the most influential of the managers of that institution. When he stopped lecturing, he continued practising as a surgeon and wrote several influential textbooks but he never forgave Gregory for exclusion from the Infirmary. He was soon recognized as one of the foremost surgeons in Scotland. He spent the last few years of his life on the Continent attending to the surgical needs of the numerous expatriate Britons who lived there. PMID- 19813309 TI - Professor Sir William Liley (1929-83): New Zealand perinatal physiologist. AB - William (Bill) Liley received his MB ChB from Otago University, Dunedin (New Zealand), in 1954. Under the guidance of the neurophysiologist Professor J C Eccles (1903-97), he carried out major research on neuromuscular transmission, both as an undergraduate at Otago University and as a postgraduate at the Australian National University at Canberra. In 1957 Bill Liley switched to research in obstetrics at the Women's National Hospital at Auckland in New Zealand. He refined the diagnostic procedure for rhesus haemolytic disease of the newborn and was able to predict its severity. Liley developed the technique of intrauterine transfusion of rhesus-negative blood for severely affected fetuses and led the team that carried out the first successful fetal transfusions in the world. He was a passionate advocate of the medical and societal rights of the unborn child. PMID- 19813310 TI - Sir Ernest Graham-Little, MP, MD FRCP (1867-1950): dermatologist and politician. AB - Sir Ernest Graham-Little was not only one of the foremost British dermatologists of the first half of the 20th century but he also served from 1924 to 1950 as an Independent Member of Parliament. As a dermatologist he played a leading role in medical education, wrote a vast number of clinical papers, was the President of both the British Association of Dermatology (as it then was) and the Section of Dermatology of the Royal Society of Medicine, and he described a form of scarring alopecia that has become known as Graham-Little syndrome. He was the first to describe the syndrome later named after Waterhouse and Friedrichsen. In Parliament he expressed his opinions on a vast array of subjects, often advancing views well ahead of his time. Perhaps most remarkably, for much of his time in Parliament he was still an active dermatologist. Notwithstanding the formidable achievements in his twin careers, his name is not well known. PMID- 19813311 TI - Joseph Landsberger (1848-1933): medical man in a time of change. AB - Joseph Landsberger was a Jewish doctor in Germany in the second half of the 19th and much of the first half of the 20th century. He was involved in the scientific advances of his time, especially in the fields of antisepsis and asepsis, bacteriology, surgical technique, public health and therapeutics. PMID- 19813312 TI - The death of Buddha: a medical enquiry. AB - The death of the Buddha (Siddhartha Gautama) has been depicted widely in Buddhist iconography. The Buddha is generally shown with a serene or smiling expression, lying on his right side and resting his head on his right hand. The dates of Buddha's life traditionally are given as 566-486 BC. Buddha died from an illness, the nature of which remains unsettled. The present paper examines a variety of sources and concludes that it was tainted pork that led to his demise. He succumbed to the disease pig-bel, a necrotizing enteritis caused by the toxins of Clostridium perfringens infection. PMID- 19813313 TI - Toxic honey and the march up-country. AB - This article gives an account of the Greek warrior-historian Xenophon and his Anabasis (The March Up-Country), one of the most famous events in military history. It includes a description of how the Greek soldiers, after reaching apparent safety near the south-eastern Black Sea, were felled by a strange honey that rendered them as if dead for a day. Modern understanding of the toxicology of this honey is given, then an epilogue summarizing the rest of Xenophon's life. PMID- 19813314 TI - Carl von Clausewitz (1780-1831) and cholera: the cause of World War II? AB - Here it is proposed that, by killing Prussian Major General Carl von Clausewitz before he could complete the work on his military text On War, cholera strongly influenced the nature of World War I (1914-18) and, by direct extension, contributed to the cause of World War II (1939-45). PMID- 19813315 TI - The portraiture of William Harvey (1578-1657) unveiled by the detective work of Geoffrey Keynes (1887-1983) and Alex Sakula (1917-2003). AB - In November 1948, Sir Geoffrey Keynes exhibited the only known portrait of William Harvey in mid-life. The recovery of this rare portrait was due to Keynes's detective work at visits to Rolls Park in Essex. Dr Alex Sakula discovered the Betchworth portraits of the Harvey family by accident when he was invited to visit a patient, Major General Goulburn, at Betchworth in Surrey. After much research work Sakula was able to mount an informative exhibition at the Royal College of Physicians. Sakula also reminded us of Eliab Harvey (1758 1830), William's descendant, captain of the Temeraire at the Battle of Trafalgar; the year 2005 marks the bicentenary. PMID- 19813316 TI - Dr Archibald Cameron (1707-53). PMID- 19813317 TI - The first woman doctor from Aberystwyth. PMID- 19813318 TI - Localized renal cell cancer: a testing ground for new approaches and technology. PMID- 19813319 TI - Guidelines update: colon and rectal cancers. PMID- 19813320 TI - Bladder cancer. Clinical practice guidelines in oncology. AB - Urothelial tumors represent a spectrum of diseases with a range of prognosis. After a diagnosis is established at any point within the urothelial tract, the patient remains at risk for developing a new lesion at a different or the same location and at a similar or more advanced stage. Continued monitoring for recurrence is an essential part of management, because most recurrences are superficial and can be managed endoscopically. Within each category of disease, more refined methods to determine prognosis and guide management, based on molecular staging, are under development with the goal of optimizing the individual patient's likelihood of cure and chance for organ preservation. For patients with more extensive disease, newer treatments typically involve combined modality approaches, using recently developed surgical procedures, or three dimensional treatment planning for more precise delivery of radiation therapy. Although these are not appropriate in all cases, they do offer the promise of an improved quality of life and prolonged survival. Finally, within the category of metastatic disease, a number of new agents have been identified that appear to be superior to those currently considered to be standard therapies. Experts believe, therefore, that the treatment of urothelial tumors will evolve rapidly over the next few years, with improved outcomes for patients at all stages of disease. PMID- 19813321 TI - Radical cystectomy and surgical quality of care. AB - Defining surgical quality is an imperative and substantial undertaking before its measurement and ultimate improvement. This article defines quality of care and a rationale for its measurement. In the context of radical cystectomy for bladder cancer, we describe a conceptual model for measuring quality of care. Finally, we provide a framework for future research by presenting an overview of recent work pertaining to cystectomy and quality of care. PMID- 19813322 TI - The integration of chemotherapy and surgery for bladder cancer. AB - Despite surgery with curative intent, approximately 50% of patients with muscle invasive transitional cell carcinoma of the bladder will develop distant metastases and succumb to their disease. Attempts to improve outcomes have focused on refining surgical techniques and integrating perioperative chemotherapy. This review summarizes the available literature addressing the role of pelvic lymphadenectomy, neoadjuvant chemotherapy, and adjuvant chemotherapy in the management of patients with muscle-invasive transitional cell carcinoma of bladder. PMID- 19813323 TI - Testicular cancer. Clinical practice guidelines. PMID- 19813324 TI - Management of advanced germ cell cancer in patients with unfavorable prognosis. AB - Advanced germ cell cancer can be cured in most patients using chemotherapy with or without surgery. A small fraction of patients with nonseminomatous tumors (NSGCT) and an even smaller percentage of seminoma patients are destined to have a less favorable outcome, due to an inadequate response to first-line chemotherapy (failure to achieve remission, finding of residual viable carcinoma at post-chemotherapy surgery, or relapse after achieving a remission). Despite the apparent salvage potential for regimens containing ifosfamide or paclitaxel, no proof exists that such combinations are superior to the standard regimen of four cycles of cisplatin, etoposide, and bleomycin (PEB) in the front-line therapy of patients with advanced NSGCT. Other modifications of first-line therapy, such as the addition of paclitaxel or the use of escalated doses of cisplatin, also have failed to increase the cure rate. The use of single or tandem cycles of high-dose chemotherapy (HDT with autologous hematopoietic cell transplant [aHCT]) in various settings (for selected patients with poor prognostic features before therapy, patients predicted to have a poor outcome based on the rate of serum tumor marker decline while on therapy, and patients in relapse or failure to achieve adequate response to standard therapy) has been evaluated in many phase II and a limited number of phase III trials, which are summarized in this review. Important questions that remain to be answered include the role of new agents and the use of more sophisticated techniques to understand prognostic and predictive factors in selecting therapy for GCT. PMID- 19813325 TI - Kidney cancer. Clinical practice guidelines. PMID- 19813326 TI - Controversies in the surgical management of renal cancer. AB - Renal tumors are a common cause of cancer, and renal cell carcinoma accounts for the vast majority of the renal tumors in the United States. The past two decades have produced numerous advances in the treatment of localized and metastatic renal cell carcinoma. Nephron-sparing surgery, laparoscopic nephrectomy, and energy-ablative techniques are now in the armamentarium of the urologist. The role of adrenalectomy and lymphadenectomy are better understood today than in decades past, and recent advances in the understanding of immunotherapy, cytoreductive nephrectomy, and metastatic disease have also improved treatment for this disease. As is often the case as technology and knowledge evolve, controversies regarding the surgical treatment of renal cancer exist. This article outlines some of these controversies and reviews the evidence surrounding each. PMID- 19813327 TI - Management of metastatic renal cancer. AB - The purpose of this article is to review the systemic management options for patients with metastatic renal cancer. We reviewed the literature regarding systemic management of metastatic renal cancer. Treatment options of chemotherapy agents, immunotherapy, molecularly targeted agents, allogeneic stem cell transplantation, vaccines, and other manipulations of the immune system are discussed. No chemotherapy agent used alone or in combination has consistently produced responses to substantiate its routine use. Interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) have shown response rates ranging from 10% to 20%. Some studies have shown that retinoids may enhance the antitumor activity of IFN alpha. Molecularly targeted agents and angiogenic agents are being actively pursued and several studies are showing response rates above 30%. Although nonmyeloablative allogeneic stem cell transplantation shows some promising results, they also have limitations to its use. Therapy strategies that incorporate vaccines as part of comprehensive immune manipulations are also being studied. The systemic treatment of patients with advanced renal cell cancer continues to be a significant challenge. Immunotherapy treatment has shown response in up to 20% of patients. Unfortunately, most do not respond. The current technologies are promising and may be the key step for introduction of better treatments for renal cancer care. PMID- 19813328 TI - [Significance of HDL cholesterol increase for the risk reduction. Methods of intervention. recommendations of experts]. PMID- 19813329 TI - [Why we support the idea that, without knowing of serum level of HDL cholesterol, diagnosis and treatment of coronary disease is difficult]. PMID- 19813330 TI - [HDL in the guidelines of cardiovascular diseases prevention]. PMID- 19813331 TI - [History and pathogenesis of metabolic syndrome]. PMID- 19813332 TI - [Metabolic syndrome as cardiovascular risk factor]. PMID- 19813333 TI - [Metabolic syndrome in Poland]. PMID- 19813334 TI - [Management of hypertension in patient with metabolic syndrome or diabetes]. PMID- 19813335 TI - [Carbohydrate metabolism disorders in metabolic syndrome]. PMID- 19813336 TI - [Treatment of lipid disorders in metabolic syndrome and type 2 diabetes. The place for fibrates]. PMID- 19813337 TI - [Acute Coronary Syndrome in the patient with metabolic syndrome and/or diabetes. Are there any differences in management?]. PMID- 19813338 TI - [Influence of ACE inhibitors and fibrates on endothelial function]. PMID- 19813339 TI - [Pleiotropic action of ACE-I and fibrates]. PMID- 19813340 TI - [Lipids disorders in chronic kidney disease]. PMID- 19813341 TI - [Combination pharmacotherapy therapy in the management of atherogenic dyslipidemia]. PMID- 19813342 TI - Planning and operational considerations for units utilizing military working dogs. AB - Military working dogs are rapidly becoming integral to military operations. While they bring many valuable capabilities to the battlefield, it is important that Special Operations leaders consider canine team capabilities and requirements when planning missions. Careful logistical and operational planning can optimize the health, performance, and readiness of the working dog while protecting the safety and well-being of the team members working with them. We also offer recommendations for medical treatment of dog bites. PMID- 19813343 TI - HIV postexposure prophylaxis for Special Forces soldiers. AB - Exposure to human immunodeficiency virus (HIV) is a recognized occupational hazard to healthcare personnel. The virus also presents an operational hazard to deployed Special Operations Forces (SOF) personnel. Management guidelines for work related exposure to HIV mainly deal with healthcare workers in a first world hospital environment. Formal guidelines for postexposure prophylaxis (PEP) regarding potential HIV exposure in third world environments have not been established. SOF personnel deploy to regions such as sub-Saharan Africa with a reported HIV prevalence of 35% or higher. This article examines the case of a SOF servicemember exposed to HIV in a confrontation with host nation personnel, the problems with trying to utilize current CDC guidelines and host-nation healthcare capabilities, and a proposed solution devised to ensure appropriate PEP in future cases. PMID- 19813344 TI - Differential diagnosis of shoulder pain followed by progressive weakness: a case report. AB - Upper extremity weakness can be the result of a myriad of conditions ranging from contractile tissue injury, joint injury, or injury to central or peripheral nervous system components. Accurate diagnosis is important in establishing an optimal treatment regimen and sound prognosis. This report provides an overview of the diagnosis and treatment of Parsonage-Turner Syndrome, a relatively rare cause of upper extremity weakness and dysfunction. PMID- 19813345 TI - Clinical red flags from the USSOCOM TMEPS--recognize, act fast, and evacuate! AB - Military medical practitioners working in active duty clinics are known for seeing lots of people in a short amount of time. If you've ever seen what goes on every morning at a troop medical clinic on a training post or base you know what I mean. The goal of morning sick call is to find that one really sick person among the many not so sick standing in line for your services. What you learn from working in that setting is how to recognize the red flag--the sign or symptom that clues you in to a potentially dangerous condition. When at war, the ability to recognize the red flag is extremely important. In the austere or unsecure environment, the SOF Medic needs to be the one who knows what the red flags are and what to do about them once spotted. PMID- 19813346 TI - The Special Operations Resuscitation Team: robust Role II medical support for today's SOF environment. AB - Special Operation Forces (SOF) have historically relied upon conventional medical assets for Role II and higher medical support. Over the last five years, the need for SOF-specific medical teams and surgical support was identified and addressed. Several Special Operations based Role II assets are now available to support operations, each with unique personnel and capabilities. The Special Operations Resuscitation Teams (SORT) have been engaged in several joint deployments in the last year, demonstrating the mission readiness and lifesaving trauma support for which the teams were designed. The future of SOF Role II has many unique challenges, including personnel resourcing, training, and joint operational planning. PMID- 19813348 TI - Ultrasound detection of pneumothorax with minimally trained sonographers: a preliminary study. AB - BACKGROUND: Prompt recognition and treatment of a tension pneumothorax is critical to reducing mortality in both military and civilian settings. Physician assistants, Special Operations Forces (SOF) and conventional force Medics are often the first medical providers to care for combat trauma patients with penetrating chest trauma and frequently have limited diagnostic capabilities available to them due to mission constraints. The purpose of this study is to examine the potential for non-physician providers to determine the absence or presence of a pneumothorax in a porcine model, with the use of a portable ultrasound machine, after receiving minimal training. METHODS: Physician assistants, SOF and conventional force Medics, veterinary technicians, and food service inspectors, all naive to ultrasound, were recruited for this study. Participants underwent a brief presentation on detection of a pneumothorax by ultrasound and were then asked to perform a thoracic ultrasound examination on euthanized, ventilated swine. Some of the swine were induced with a pneumothorax prior to these examinations, and all participants were blinded to the absence or presence of a pneumothorax. RESULTS: Twenty-two participants examined a total of 44 hemithoraces. A total of 21 out of 22 pneumothoraces were correctly identified with one false-negative. All 22 normal hemithoraces were correctly identified for a sensitivity of 95.4% (95% CI 0.75-0.99), and a specificity of 100% (95% CI 0.81 1.00), with PPV of 100%, NPV of 95.6%. CONCLUSIONS: Non-physician healthcare providers can accurately detect a pneumothorax with portable ultrasound after receiving minimal focused training. PMID- 19813347 TI - Field evaluation and management of non-battle related knee and ankle injuries by the advanced tactical practitioner in an austere environment. Part one. PMID- 19813350 TI - An evaluation of tactical combat casualty care interventions in a combat environment. AB - BACKGROUND: Tactical combat casualty care (TCCC) is a system of prehospital trauma care designed for the combat environment. Although widely adopted, very few studies have reported on how TCCC interventions are actually delivered on the battlefield, from a quality of care perspective. STUDY DESIGN: This was a prospective study of all trauma patients treated at the Role 3 multinational medical unit (MMU) at Kandahar Airfield Base from February 7, 2006 to May 30, 2006. Primary outcomes were whether or not two TCCC interventions were underused, overused, or misused. Interventions studied were needle decompression of tension pneumothoraces and tourniquet application for exsanguinating extremity injuries. RESULTS: One hundred thirty-four trauma patients were treated at the Role 3 MMU during the study period. Six patients had eight tourniquets applied. Five tourniquets were applied to four patients appropriately and saved their lives. There was one case of misuse where a venous tourniquet was applied. There was one case of overuse where one patient had two tourniquets placed for 4 hours on extremities with no vascular injury. There were seven cases where needle decompression was underused: Seven patients presented with vital signs absent with no needle decompression. There was one case of overuse of needle decompression. There were seven cases of misuse where the patients were decompressed too medially. CONCLUSIONS: Tourniquets save lives. Needle decompression can save lives, but is usually performed in patients with multiple critical injuries. TCCC instructors must reinforce proper techniques and indications for each procedure to ensure that the quality of care provided to injured soldiers on the battlefield remains high. PMID- 19813349 TI - Tourniquets for the control of traumatic hemorrhage: a review of the literature. PMID- 19813351 TI - The command of biotechnology and merciful conquest in military opposition. AB - Biotechnology has an increasingly extensive use for military purposes. With the upcoming age of biotechnology, military operations are depending more on biotechnical methods. Judging from the evolving law of the theory of command, the command of biotechnology is feasible and inevitable. The report discusses some basic characteristics of modern theories of command, as well as the mature possibility of the command theory of military biotechnology. The evolution of the command theory is closely associated with the development of military medicine. This theory is expected to achieve successes in wars in an ultramicro, nonlethal, reversible, and merciful way and will play an important role in biotechnological identification and orientation, defense and attack, and the maintenance of fighting powers and biological monitoring. The command of military biotechnology has not become a part of the virtual military power yet, but it is an exigent strategic task to construct and perfect this theory. PMID- 19813352 TI - Tourniquet use in combat trauma: U.K. military experience. AB - AIM: To determine the prevalence of tourniquet use in combat trauma, the contribution to lives saved and the complications of their use in this environment. POPULATION: All casualties treated at U.K. field hospital facilities in Iraq and Afghanistan and meeting criteria for entry into U.K. Joint Theatre Trauma Registry (JTTR) from 04 Feb 03 to 30 Sep 07. METHODS: Cases were identified from U.K. JTTR. Casualties from Permanent Joint Overseas Bases (PJOBs) were excluded. ISS, NISS, TRISS and ASCOT were calculated automatically within JTTR from AIS 2005 (Military) codes. RESULTS: 1375 patients met U.K. JTTR entry criteria for the period specified (excluding PJOBs). 70/1375 patients (5.1%) were treated with one or more tourniquets (total 107 tourniquet applications). 61/70 (87%) survived their injuries. 17/70 (24%) patients had 2 or more tourniquets applied. 64/70 patients received a tourniquet after April 2006, when tourniquets were introduced as an individual first aid item. 43/70 (61%) patients were U.K. military. CONCLUSIONS: ISS and TRISS are poorly representative of injury severity and outcome for combat trauma involving isolated multiple limb injuries and cannot be used to discriminate whether a tourniquet is life-saving. The presence of severe isolated limb injuries, profound hypovolaemic shock and the requirement for massive transfusion reasonably identifies a cohort where the use of one or more tourniquets pre-hospital to control external bleeding can be said to be life saving. PMID- 19813353 TI - Company command building: combat-ready teams--making sense of killing. PMID- 19813354 TI - Letter to the USSOCOM/SG education and training section. PMID- 19813355 TI - Picture this...Onchocerciasis. PMID- 19813356 TI - What's happening in midwifery education? PMID- 19813357 TI - Universities are already providing successful breastfeeding training. PMID- 19813358 TI - Cover should reflect content. PMID- 19813359 TI - The development of maternity care assistants in Scotland. PMID- 19813360 TI - Giving birth to the maternity care assistant. PMID- 19813361 TI - 'She gave me the confidence': positive mentoring. PMID- 19813362 TI - In praise of mentors. PMID- 19813363 TI - Learning from the 'stork nurse'. PMID- 19813364 TI - Still a midwife? The transition from clinician to educator. PMID- 19813365 TI - Going for the dream: the ups and downs of application. PMID- 19813366 TI - Emotional freedom technique (2). PMID- 19813367 TI - Inspiration from the past (1): Jane Sharp. PMID- 19813368 TI - Experiences of early labour (2): strategies for coping at home. PMID- 19813369 TI - Visiting Osani. PMID- 19813370 TI - Screening: what are we missing? PMID- 19813371 TI - Forgetting how to care. PMID- 19813372 TI - Ward leaders need authority if they are to meet their responsibilities. PMID- 19813373 TI - Preparation, training and support: the cornerstones of ward leadership. PMID- 19813374 TI - The same, but different. AB - With more nursing students opting for degree level entry to the profession, the demand for broad experience is growing. One solution is an exchange programme. PMID- 19813375 TI - Every picture tells a story. AB - Children can be psychologically, as well as physically, damaged by meningitis. Art therapy can help them to explore their fears. PMID- 19813377 TI - Show you care. PMID- 19813376 TI - A conflict of principles? AB - Nurses in countries with laws enabling assisted suicide have a wide range of views. PMID- 19813378 TI - A shot in the dark. PMID- 19813379 TI - Osteoarthritis: pathogenesis, clinical features and management. AB - Osteoarthritis is a degenerative disorder of the synovial joints that is characterised by joint pain, stiffness and limited range of movement. The majority of nurses will encounter patients with this condition even though this might not be the reason they are seeking health care. This article discusses the associated signs and symptoms, investigations, diagnosis and treatment of this condition. PMID- 19813380 TI - Implementation of the NHS Knowledge and Skills Framework. AB - This article describes the implementation of the NHS Knowledge and Skills Framework (KSF) by part of an NHS trust that provides services for people with learning disabilities. It highlights factors that assisted and hindered the implementation of the KSF across services. PMID- 19813382 TI - Literature review. PMID- 19813381 TI - Treatment of hyperglycaemia in patients with type 2 diabetes. AB - This article provides an overview of the lifestyle advice and drug treatment of hyperglycaemia in adults with type 2 diabetes. The value of patient education is highlighted, as is the importance of working with patients to set relevant goals. Long-term complications are briefly discussed along with the multi-faceted treatment of type 2 diabetes, which includes management of blood pressure, lipids and other cardiovascular risk factors in addition to hyperglycaemia. PMID- 19813383 TI - Mutually supportive. PMID- 19813384 TI - Making the most of careers fairs. PMID- 19813385 TI - In search of stones, bones, abdominal groans and psychic moans: putting the pieces together--a case of parathyroid adenoma. AB - An often neglected and frequently forgotten gland in the body is the parathyroid. The parathyroid glands (there are four) play a vital role in maintaining body function by regulating calcium levels through the secretion of parathyroid hormone. When function of the parathyroid gland becomes abnormal and calcium regulation is altered, there can be serious consequences to the patient in the form of a parathyroid adenoma. There is an increased incidence in females older than sixty-five. Due to the variable symptoms, making an accurate diagnosis can be a challenge for the clinician. This paper will provide an overview of the parathyroid glands, identification of the signs and symptoms of a parathyroid adenoma, a discussion about the tests that facilitate a definitive diagnosis and a review of treatment options. A Case Study will assist in putting all the pieces together. PMID- 19813386 TI - Resident centred decision-making and end-of-life treatment in long-term care: what are the consequences to the elderly patient? PMID- 19813387 TI - My journey into gerontological nursing: we are each on unique journeys as gerontological nurses. PMID- 19813388 TI - Strength through unity. PMID- 19813389 TI - Stop dancing with the stars: why the 6-month boogie is costing you money and freedom. PMID- 19813390 TI - How to win in a tough economy with superior customer service. PMID- 19813391 TI - Young adults with disabilities and uninsured for health care. PMID- 19813392 TI - Create. PMID- 19813393 TI - Medicine's NFL draft. PMID- 19813394 TI - Did we drop the ball? PMID- 19813395 TI - ACH fitness clinic. Combating pediatric obesity in clinic and through teaching. PMID- 19813396 TI - Healthcare-associated infections: Arkansas hospitals work to improve patient care and cut costs through prevention. PMID- 19813397 TI - Case of the missed opportunity for temporal artery biopsy. PMID- 19813398 TI - Legally speaking: Federal Trade Commission issues positive advisory opinions on antitrust implications of clinical integration. AB - From time to time, groups of physicians in an area may determine that they would benefit from "integrating" their practices into an IPA, PHO, or other joint venture. The anticipated benefits may include economies of scale, the ability to coordinate care between primary care physicians and specialists, providing disease management services for patients with certain conditions, or a myriad of other reasons. A key characteristic of these proposed integrated models is the ability for the group as a whole to negotiate with insurance companies and self funded health care plans. When a group reaches the point of negotiating collectively for the fees that a pay- or is going to pay for various services throughout the plan, possible antitrust implications arise. PMID- 19813399 TI - A better future for children with type 1 diabetes: Review of the conclusions from the Diabetes Control and Complications Trial and the Epidemiology of Diabetes Interventions and Complications study. AB - Research shows a causal association between hyperglycemia and diabetic complications which include nephropathy, neuropathy, retinopathy and blood vessel and heart disease. Intensive control of blood glucose in children has been debated for years. The Diabetes Control and Complications Trial (DCCT) and the Epidemiology of Diabetes Interventions and Complications (EDIC) study followed more than 1,400 volunteers and intensely managed a portion of those patients over several years. Results from the DCCT study regarding the benefits of intensive diabetes management continue to be significant more than 15 years after they were published. PMID- 19813400 TI - Nurses put politicians on the spot over health as general election looms. PMID- 19813401 TI - Inspiring leaders. AB - Ward sisters and charge nurses need managerial and financial support to provide strong leadership and perform their roles to best effect. PMID- 19813402 TI - Blooming marvellous. AB - Staff at a rehabilitation unit have transformed an unappealing space into a therapeutic garden where patients can improve their physical and mental health. PMID- 19813403 TI - Team health and safety. AB - Patient Safety First is a campaigning organisation dedicated to reducing patient harm. It has adopted a safe surgery checklist that is being implemented across England. PMID- 19813404 TI - History repeats. PMID- 19813405 TI - A technological headache. PMID- 19813406 TI - Evaluating the impact of a cardiac module for post-registration nurses. AB - AIM: To discover whether nursing students believed that a post-registration cardiac module was meeting their needs. METHOD: A descriptive qualitative approach was used in this study. Semi-structured interviews were recorded with a purposive sample of six registered nurses who had previously completed a cardiac module. FINDINGS: Five core themes were identified: the value of the module, being a student, thinking about practice, improving delivery of care and role expansion. CONCLUSION: The study explored the extent to which a cardiac module developed the nurses' confidence, skills and knowledge in the context of their ongoing careers. The findings were generally positive because the nurses recognised the benefits for their practice and personal development. PMID- 19813407 TI - The nurse's role in helping older people to use the toilet. AB - Going to the toilet to eliminate bodily waste in private is a normal daily activity that people tend to take for granted. The inability to perform this task independently can result in an intrusive, embarrassing and stressful experience for the individual. This article outlines the nurse's role in helping older people to use the toilet and the opportunities that this affords for therapeutic intervention, identification of other problems and the promotion of health and wellbeing. PMID- 19813408 TI - Assessing and treating faecal incontinence in children. AB - Faecal incontinence can have a profound effect on the lives of children and their families. Children who have faecal incontinence have a greater risk of being bullied at school, and parents are often frustrated and concerned by the associated social stigma. The social and psychological effects of faecal incontinence on the child can last for a long time. This article provides an overview of the causes of faecal incontinence, discusses assessment of bowel dysfunction and outlines current treatments. The article also highlights the importance of the nurse's role, as part of the multidisciplinary team, in assessing, treating and supporting children and their families to ensure that any interventions have the best chance of succeeding and to minimise the risk of relapse. PMID- 19813410 TI - Study by degrees. PMID- 19813409 TI - Stab wounds. PMID- 19813411 TI - The neighbourhood revolution. PMID- 19813412 TI - Obama's vision. PMID- 19813413 TI - Medicine, conflict and survival: change and continuity. PMID- 19813414 TI - From Solferino to Sri Lanka: health workers, health information, and international law. PMID- 19813415 TI - Call for a global response 2010: new Internet resource and forthcoming conference to examine the links between violent conflict and health. PMID- 19813416 TI - Childhood cancers near German nuclear power stations: the ongoing debate. AB - In late 2007, the significant KiKK study (Kinderkrebs in der Umgebung von KernKraftwerken = Childhood Cancer in the Vicinity of Nuclear Power Plants) in Germany reported a 1.6-fold increase in all cancers and a 2.2-fold increase in leukaemias, among children living within 5 km of all German nuclear power stations. The KiKK study by Kaatsch et al. was extensively described in a recent edition of Medicine Conflict and Survival. It has triggered much discussion as to the cause(s) of these increased cancers. This article reports on recent developments on the KiKK study, including responses by German radiation agencies, and recent epidemiological studies near United Kingdom and French nuclear installations. It reflects the current debate and concludes with advice to policy makers on radiation risks on the relative merits of the KiKK study. An accompanying article outlines a possible explanation for the increased cancers and makes recommendations for future research. PMID- 19813417 TI - Childhood cancers near German nuclear power stations: hypothesis to explain the cancer increases. AB - In early 2008, the very large Kinderkrebs in der Umgebung von Kernkraftwerken [Childhood Cancer near Nuclear Power Plants] (KiKK) study in Germany reported increases in leukaemias and solid cancers among children living near all German nuclear power plants (NPPs). This study, previously described in Medicine, Conflict and Survival, has triggered debates in many countries as to the cause or causes of these increased cancers. An accompanying article reports on the recent developments on the KiKK study including the responses by German radiation agencies, and the results of recent epidemiological studies near United Kingdom and French nuclear installations. This article outlines a possible explanation for the increased cancers. In essence, doses from environmental NPP emissions to embryos/foetuses in pregnant women near NPPs may be larger than suspected, and haematopoietic tissues may be considerably more radiosensitive in embryos/foetuses than in newborn babies. The article concludes with recommendations for further research. PMID- 19813418 TI - Footprints to the future. AB - A brief history of the impact of humans on the earth's biosphere and a description of the more serious of these lead into a description of a method of quantifying the overall contemporary impact. The world's available biological resources--assessed by global footprinting data published by the Global Footprinting Network in October 2008--are compared with the demand for more and more energy by the high, medium and low income group countries. The inescapable conclusion is that, without fossil fuels, the planet cannot support the current population of 6.8 billion, except at significantly lower average levels of consumption. As the population is projected to grow to 9.4 billion by 2050, by which time available energy and earth's biocapacity will have diminished due to the adverse effects of global warming, there is an urgent case for encouraging people to limit family sizes to two or fewer children to avoid the worst effects of the intrinsic resource-consumption mismatch. Examples of countries which have already recognized the problem nationally are described to show that such actions are not only possible and successful, but are becoming increasingly necessary. PMID- 19813419 TI - A matter of time and place: violent conflict and civilian physical health. PMID- 19813420 TI - Sinus grafting with a natural fluorohydroxyapatite for immediate load: a study with histologic analysis and histomorphometry. AB - The goal of this retrospective study was to evaluate the survival rates of dental implants placed in sinuses grafted with a 50:50 composite ratio of autogenous bone and a natural flourohydroxyapatite (FHA) combined with platelet-rich plasma (PRP) using an immediate-load protocol. The authors hypothesized that a 50:50 composite ratio of FHA and autogenous bone combined with PRP would permit immediate loading without compromising implant survival rates. Eleven patients with bilateral partial edentulism of the posterior maxilla were enrolled in this retrospective study. Autogenous bone used in the graft procedure was harvested from the tibia of the left lower extremity. Each patient was grafted with a 50:50 composite ratio of autogenous bone and FHA. Membranes were not used to cover the lateral wall osteotomy site. Platelet-rich plasma was added to the graft material to accelerate and enhance bone regeneration. Four to 6 months after the grafting procedure, 37 hydroxyapatite-coated dental implants were surgically placed and immediately loaded between 72 hours and 5 days later with custom titanium abutments and acrylic provisional restorations placed out of functional occlusion. Six months later, definitive ceramometal restorations were cemented on to the custom abutments. Patients were observed over a 52-week period. The overall implant survival rate was 97.3%. Histologic and histomorphometric analysis of core samples revealed formation of new vital bone in different graft specimens ranging from 23% to 34%. In each core bone sample, 100% of the bone sample was determined to be vital. In the grafted maxillary sinus, the natural FHA combined with autogenous bone in a 50:50 composite ratio with PRP is a suitable graft material permitting immediate load without compromising implant survival rates while decreasing the overall healing time. PMID- 19813421 TI - CaPO4 blasted implants inserted into iliac crest homologue frozen grafts. AB - In the last decade, some investigations have reported that the resorbable blast media surface (also named CaPO4 blasted implants [CaPO4-Bls]) has achieved excellent results. However, no report regarding CaPO4-Bls inserted into fresh frozen bone (FFB) is available. Thus, we planned a retrospective study on a series of CaPO4-Bls inserted into FFB to evaluate their clinical outcome. In the period between December 2003 and December 2006, 16 patients (10 females and 6 males, median age of 55 years) were operated on, and 76 CaPO4-Bls were inserted. The mean implant follow-up was 23 months. Implant diameter and length ranged from 3.25 to 4.5 mm and from 11.5 to 15 mm, respectively. Implants were inserted to replace 7 incisors, 11 cuspids, 31 premolars, and 27 molars. Only 1 out of 76 implants was lost (i.e., survival rate [SVR] = 98.7%), and no differences were detected among the studied variables. When peri-implant crestal bone resorption was used as an indicator of clinical success (i.e., success rate), it was possible to identify some variables that correlated with a better clinical outcome. Specifically, Cox regression showed that removable prosthetic restoration and longer implant length correlated with a statistically significant lower delta implant abutment junction (IAJ; i.e., reduced crestal bone loss) and thus a better clinical outcome. In this study, CaPO4-Bls had high survival and success rates, similar to those reported in previous reports of 2-stage procedures in nongrafted bone. CaPO4-Bls inserted into FFB are reliable devices, although greater marginal bone loss occurs when fixed prosthetic restorations and short implants are used. PMID- 19813422 TI - Implant success in distracted bone versus autogenous bone-grafted sites. AB - Endosseous implants are the treatment of choice for restoring function and reconstructing most edentulous areas of the maxilla and mandible. In general, alveolar bone defects can be reconstructed by either distraction osteogenesis or autogenous bone grafting. After alveolar reconstruction, endosseous implants are used to support and retain the prosthesis for restoration of form and function. Eighty-two consecutive patients requiring alveolar augmentation prior to implant placement were evaluated. All patients were given treatment options for reconstructing their alveolar defects, which included autogenous bone grafting vs distraction osteogenesis. Sixty-five patients received autogenous grafts (anterior iliac crest: 44; retromolar: 17; tibia: 2; chin: 2), and 17 patients underwent distraction osteogenesis prior to implant placement. A total of 184 implants were placed in the autogenous bone-grafted sites and 56 implants in the distracted bone sites. Implants placed in sites restored with autogenous bone grafts had an implant success rate of 97% (178/184), whereas implants placed in distracted bone sites had a success rate of 98% (55/56). In the autogenous grafted group, 3 implants failed in the posterior mandible, one in the anterior maxilla, one in the anterior mandible, and one in the posterior maxilla. In the distraction group, one implant failed in the posterior mandible. Both techniques are associated with good success rates. There was no statistical difference between implant success in autogenous bone vs distracted bone sites in this group of patients. PMID- 19813423 TI - Measurement of clinicians' ability to hand torque dental implant components. AB - There is a varying degree of hand torque abilities using finger drivers among clinicians. Calibrating one's own abilities requires complicated instruments not readily available. This study evaluated a simple-to-use method that allows dental practitioners to have a quantifiable clinical assessment of relative torque ability using finger drivers to torque down dental implant components. A typodont that includes dental implants was mounted in a mannequin placed in a patient reclined position. The subjects were asked to torque as tightly as they could a new healing abutment to an implant secured firmly in resin within the typodont. All participants wore moistened gloves when using a finger driver. The healing abutment was countertorqued using a certified precalibrated precision torque measurement device. The reading on the torque driver was recorded when the healing abutment disengaged. An average of torque values of dentists and dental students was calculated. Fifty subjects had an average maximum torque ability of 24 Ncm (male dentists: 28 Ncm; students: 22 Ncm; male students: 24 Ncm; female students: 19 Ncm). Maximum torque values for all participants ranged from 11 Ncm to 38 Ncm. There was no significant difference between groups. This study showed a varying degree of hand torquing abilities using a finger driver. Clinicians should regularly calibrate their ability to torque implant components to more predictably perform implant dentistry. Dental implant manufacturers should more precisely instruct clinicians as to maximum torque, as opposed to "finger tighten only". PMID- 19813424 TI - Immediate implant placement after removal of a failed implant: a clinical and histological case report. AB - The purpose of this study was to evaluate the clinical success of an implant placed immediately after the explantation of a fractured blade implant. A healthy 58-year-old male nonsmoker presented with a fractured blade implant that had been subjected to biomechanical overload. A new blade implant was placed immediately after the removal of the fractured one. The new implant was placed with a composite graft of collagen gel and corticocancellous porcine bone and covered with a bioabsorbable membrane. Radiographic evaluation at 6 months postoperation showed complete bone healing. No residual bone defect was observed or probed during the uncovering phase; moreover, no mobility, pain, suppuration, or presence of peri-implant radiolucency were observed at the second-stage surgery. PMID- 19813425 TI - Effect of implants on maximum bite force in edentulous patients. AB - One of the main goals of prosthetic dentistry is to reconstruct the masticatory system. Replacing missing teeth by complete or partial denture is associated with decreased bite force and patient satisfaction. The aim of the present study was to measure the maximum bite force (MBF) and to evaluate patients' satisfaction from their conventional complete dentures and their mandibular implant-supported overdentures opposed by complete denture. In this cross sectional analytical descriptive study, seventy-five 45- to 65-year-old patients were divided into 3 groups wearing: (1) conventional complete dentures for up to 6 months; (2) conventional complete dentures for 10 years or more; and (3) complete maxillary dentures opposing mandibular implant-supported overdentures. Bite force was measured by means of electronic bite force measuring device with strain gauges. Three measurements were made on each side on the first molar region and the mean values were recorded. Patient satisfaction was also recorded using questionnaires. Results were analyzed by analysis of variance (ANOVA), t test, chi-square, and Pearson correlation test. The mean MBF for the first, the second, and the third group was, respectively, 5.65 +/- 1.46 kgf, 7.01 +/- 2.1 kgf, and 12.22 +/- 27 kgf. The difference between MBF in patients with overdentures and in patients with conventional complete dentures was statistically significant (P = .001). Patient satisfaction with mandibular implant-supported overdentures was recorded as "great." Results indicated MBF in the patients with mandibular implant-supported overdentures was significantly higher than that of patients with conventional complete dentures. This difference may indicate the important role of dental implants in the improvement of bite force and chewing efficiency and therefore patients' satisfaction. PMID- 19813426 TI - A technique to modify the length of an implant healing abutment. AB - In areas where esthetics is paramount, healing should be adequately completed around the implant healing abutment to stabilize the gingival margin before crown fabrication. This article describes a technique to increase the height of the healing abutment with laser welding when an implant is placed deep with thick overlying soft tissue. PMID- 19813427 TI - Calcium hydroxide paste as a surface detoxifying agent for infected dental implants: two case reports. AB - Dental implant treatment is successful; however, an implant can become infected during or after osseointegration. The two case reports presented here demonstrate, anecdotally, the effectiveness of endodontic calcium hydroxide paste for the surface treatment of infected, healing, or osseointegrated dental implants. Calcium hydroxide may be an appropriate surface detoxifying agent for local dental implant infections. A sequence of calcium hydroxide and 0.2% chlorhexidine gluconate surface treatment may provide a broad range of antimicrobial action for detoxifying recalcitrant infections. Calcium hydroxide should not be left in the surgical site. PMID- 19813428 TI - Management of dental implant fractures. A case history. AB - The widespread use of endosseous osseointegrated implants to replace missing natural teeth increases the chances of implant complications and failures, despite the high initial success rate reported in the literature. Implant fracture is one possible complication that results in ultimate failure of the dental implant. Such a complication poses a management crisis even for the most experienced clinician. This article reports on a case of implant fracture, its possible causes, and how the case was managed. PMID- 19813429 TI - KMA Legal Trust Fund protects physicians' rights. PMID- 19813430 TI - Parental vaccine concerns in Kentucky. AB - BACKGROUND: An increasing number of parents are questioning the safety and necessity of routine childhood immunizations. Locally produced vaccine risk communication materials may be effective in reassuring these parents. However, little is known about specific vaccine safety concerns in the state of Kentucky. METHODS: An Internet-based survey focusing on parental vaccine safety concerns and potential vaccine risk communication strategies was sent to all members of the Kentucky Chapter of the Amerian Academy of Pediatrics. RESULTS: There were 121 respondents who routinely administered childhood vaccines. Of these, 85% reported parental concern about the combined measles-mumps-rubella (MMR) vaccine. Concerns about the influenza and human papillomavirus (HPV) vaccines were also frequent. Of the respondents, 46% noted parental skepticism about all vaccines in general. However, refusal of all vaccines was uncommon in most practices (median 1%, interquartile range 1%-3%). The belief that vaccines cause autism was the most prevalent parental concern, reported by 70% of pediatricians. Physicians also reported that a list of reliable vaccine information Websites and pamphlets addressing common vaccine safety concerns would be the most helpful materials to use during their discussions with concerned parents. CONCLUSION: These findings suggest that specific information about the MMR, influenza, and HPV vaccines, as well as data refuting the putative link between vaccines and autism would be useful to physicians who administer vaccinations. Respondents were especially interested in reliable vaccine information on the Internet. The Websites listed below offer accurate scientific information about vaccines and the diseases they prevent. PMID- 19813431 TI - Cornelia de Lange Syndrome: a case report with clinical review and recommended anticipatory guidance for the general practitioner. AB - Cornelia de Lange Syndrome is a rare congenital malformation syndrome with typical craniofacial abnormalities and can affect the musculoskeletal, cardiovascular, gastrointestinal, genitourinary, and central nervous systems. Not all the patients look alike as the phenotypic appearance is seen on a spectrum. A classic case of CdLS has been reviewed in this article and we have discussed diagnosis, management, and anticipatory guidance. Maintaining consistent health supervision visits and outpatient therapy is vital for these children. Referring the parents to a genetic counselor is recommended if the family desires to have more children. The CdLS foundation Webpage is www.cdlsusa.org. PMID- 19813433 TI - Employed physicians. Report of the Employed Physicians Task Force. PMID- 19813432 TI - Increasing the medical school applicant pool: a key to training more rural physicians. AB - CONTEXT: Workforce studies show shortages of physicians in many areas of the United States. These shortages are especially severe in states such as Kentucky with many rural counties and are predicted to worsen in the future unless there are changes throughout our educational system to build aspirations and prepare students for medical school education. PURPOSE: To examine rural-urban differences and community characteristics of applicants and matriculants to Kentucky's two allopathic medical schools and influences on the educational aspirations of young students who wish to become physicians. METHODS: The number of Kentucky applicants and matriculants to allopathic medical schools was obtained from the Association of American Medical College's data warehouse for the period from 2002-2006. A continuous, multidimensional measure was used to classify counties by degree of rurality. Socio-demographic variables were selected for the counties of residence for applicants and matriculants. Model variables were tested in a least squares multiple regression model for their ability to explain patterns among Kentucky's 120 counties in the number of both resident applicants and matriculants to medical school. Data from a survey of middle school participants in summer health camps were analyzed to help identify important influences on young students aspiring to a career as a health professional, especially becoming a physician, and how these might be supported to increase the supply of rural medical school applicants. FINDINGS: The low number of rural applicants to medical school was highly correlated with the relative rurality of their county of residence, a low physician-to-population ratio and a low number of total primary care physicians. The percentage of county residents having a bachelor's degree level of education or higher had a positive impact on the application rate. Respondents became interested in health careers at age 15 or younger, and parents and grandparents, teachers, and close associates stimulated their aspirations, with teachers being the most influential. CONCLUSIONS: Prospective students respond to their perception of need for physicians. Rural students are influenced by those who are more highly educated. To overcome the shortage of physicians in rural communities efforts must be made to increase the aspirations for medical education of prospective students from rural counties. PMID- 19813434 TI - Comments on online publication. PMID- 19813435 TI - An update on mammography use in Canada. AB - BACKGROUND: This article updates mammography use by Canadian women aged 50 to 69, and reports trends from 1990 to 2008 among the provinces. Characteristics of non users are examined. DATA SOURCES AND METHODS: Data from the 2008 Canadian Community Health Survey (CCHS) were used to update mammography use and to examine factors associated with non-use. Historical estimates were produced using the 2000/2001,2003 and 2005 CCHS, the 1994/1995, 1996/1997 and 1998/1999 National Population Health Survey and the 1990 Health Promotion Survey. Frequency estimates, cross-tabulations and logistic regression analysis were used. RESULTS: In 2008, 72% of women aged 50 to 69 reported having had a mammogram in the past two years, up from 40% in 1990. The increase occurred from 1990 to 2000/2001; rates then stabilized. Between 1990 and 2000/2001, the difference in participation between women in the highest and lowest income quintiles gradually narrowed-from a 26- to a 12-percentage-point difference. In 2008, the disparity widened to 18 percentage points. Non-use was high in British Columbia, Prince Edward Island and Nunavut. Non-use was associated with being an immigrant, living in a lower income household, not having a regular doctor and smoking. PMID- 19813436 TI - Colorectal cancer testing in Canada--2008. AB - OBJECTIVES: This article provides estimates of the report level of colorectal cancer (CRC) testing in the Canadian population aged 50 or older in 2008. DATA SOURCES AND METHODS: The data are from the 2008 Canadian Community Health Survey. With weighted data, the percentage of people who had undergone CRC testing (fecal occult blood test in the past two years or endoscopy within the past five years) was estimated. Bivariate and multivariate analyses were used to examine testing status in relation to personal, socio-economic and other health-related characteristics. RESULTS: In 2008, an estimated 40% of Canadians aged 50 or older reported that they had had CRC testing. The percentage ranged from 28% in Quebec to 53% in Manitoba. Testing was associated with being 65 or older, higher income, having a regular doctor, being a non-smoker, and being physically active. INTERPRETATION: Organized CRC screening was limited in 2008, but may account for some of the differences in participation among the provinces. PMID- 19813437 TI - Smoking cessation: intentions, attempts and techniques. AB - BACKGROUND: A better understanding of charateristics associated with intentions to quit smoking and quit attempts and of the use of various aids is required to develop effective cessation strategies. DATA AND METHODS: Data from the 2006 Canadian Tobacco Use Monitoring Survey were used to examine intentions to quit smoking, quit attempts, use of cessation aids, and receipt of cessation advice from health professionals. RESULTS: One-third of Canadian smokers aged 15 or older reported intentions to quit within the next 30 days, and almost half had tried to quit in the past year. The number of cigarettes smoked per day was associated with intending to quit and quit attempts. The intention to quit was strongly associated with the number of past quit attempts. Half of current smokers who had seen a doctor in the past year had been advised to reduce or quit smoking. PMID- 19813438 TI - Diet quality in Canada. AB - BACKGROUND: In addition to recommendations about the consumption of specific foods and nutriens, a measure of overall diet quality is useful. Over the years, a number of countries, but not Canada, have developed indexes to evaluate diet quality. DATA AND METHODS: The American Healthy Eating Index was adapted to conform to recommendations in Canada's Food Guide. Data from 33,664 respondents to the 2004 Canadian Community Health Survey-Nutrition were used. Usual index scores were calculated with the Software for Intake Distribution Estimation program. Multiple linear regression models were used to examine associations between index scores and various characteristics, particularly the frequency of vegetable and fruit consumption. RESULTS: For the population aged 2 or older, the average score on the Canadian adaptation of the Healthy Eating Index in 2004 was 58.8 out of a possible 100 points. Children aged 2 to 8 had the highest average scores (65 or more). Average scores tended to fall into early adolescence, stabiilizing around 55 at ages 14 to 30. A gradual upturn thereafter brought the average score to around 60 at age 71 or older. At all ages, women's scores exceeded those of men. The frequency of vegetable and fruit consumption was linked to index scores. INTERPRETATION: The American Healthy Eating Index can be adapted to Canadian food intake recommendations. Canadian Community Health Survey questions about the frequency of vegetable and fruit consumption can be used as an approximation of diet quality. PMID- 19813439 TI - Health status, preventive behaviour and risk factors among female nurses. AB - BACKGROUND: This study compares the health status, preventive behaviour and risk factors of female nurses with those of other employed postsecondary-educated women. DATA AND METHODS; Cross-sectional data from the 2003 Canadian Community Health Survey were analyzed. Multiple logistic regression analyses were conducted to adjust for potential confounding by demographic and socio-economic characteristics. RESULTS: When confounding by demographic and socioeconomic characteristics was taken into account, nurses were more likely than other employed postsecondary-educated women to report back problems, that most work days were "quite a bit" or "extremely" stressful, and having had flu immunizations and cervical cancer screening. They were less likely to report insufficient consumption of vegetables and fruit or heavy alcohol use. INTERPRETATION: Canadian nurses' occupation may account for their higher prevalence of back problems and work stress. At the same time, their occupation may motivate flu immunization, cervical cancer screening, and vegetable and fruit consumption. Some problematic aspects of nurses' health profile are similar to those of other educated women. PMID- 19813441 TI - Taking the tension out of portal hypertension. PMID- 19813440 TI - Developmental pathways leading to obesity in childhood. AB - BACKGROUND: Researchers have yet to provide a comprehensive explanation of the variability in the development of childhood obesity, owing in part to the dearth of longitudinal studies. Such an understanding would contribute to the improvement of approaches for the primary and secondary prevention of childhood obesity. This study identifies, in a representative sample of Canadian children, age-related patterns of overweight and obesity between toddlerhood and childhood. DATA AND METHODS: The data are from cycles 2 through 5 (1996/1997 to 2002/2003) of the National Longitudinal Survey of Children and Youth. The sample comprised children aged 24 to 35 months at baseline, who were followed biennially over six years. Group-based mixture modelling analyses (using SAS PROC TRAJ) were conducted to identify the sex-specific developmental trajectories of body mass index (BMI). RESULTS: Group-based modelling identified four BMI trajectories for the girls (stable normal BMI, early declining BMI, late declining BMI, and an accelerating rise to obesity) and three for the boys (stable normal BMI, transient high BMI, and a J-curve rise to obesity). INTERPRETATION: Identifying distinct, sex-specific BMI trajectories is valuable in understanding pathways through which a child may develop obesity. These findings have implications for further research and practice, in particular, that no single approach can be used to prevent or reduce levels of obesity. PMID- 19813442 TI - Early rebleeding and death at 6 weeks in alcoholic cirrhotic patients with acute variceal bleeding treated with emergency endoscopic injection sclerotherapy. AB - BACKGROUND: This study evaluated the incidence of rebleeding and death at 6 weeks after a first episode of acute variceal haemorrhage (AVH) treated by emergency endoscopic sclerotherapy in a large cohort of alcoholic cirrhotic patients. METHODS: From January 1984 to December 2006, 310 alcoholic cirrhotic patients (242 men, 68 women; mean age 51.7 years) with AVH underwent 786 endoscopic variceal injection treatments (342 emergency, 444 elective) during 919 endoscopy sessions in the first 6 weeks after the first variceal bleed. Endoscopic control of initial bleeding, variceal rebleeding and survival at 6 weeks were recorded. RESULTS: Endoscopic intervention controlled AVH in 304 of 310 patients (98.1%). Seventy-five patients (24.2%) rebled, 38 (12.3%) within 5 days and 37 (11.9%) within 6 weeks. No patient scored as Child-Pugh A died. Seventy-seven (24.8%) Child-Pugh B and C patients died, 29 (9.3%) within 5 days and 48 (15.4%) between 6 and 42 days. Mortality increased exponentially as the Child-Pugh score increased, reaching 80% when the score exceeded 13. CONCLUSION: Despite initial control of variceal haemorrhage, 1 in 4 patients (24.2%) rebled within 6 weeks. Survival at 6 weeks was 75.2% and was influenced by the severity of liver failure, with most deaths occurring in Child-Pugh grade C patients. PMID- 19813443 TI - Selective observational management of penetrating neck injury in northern Nigeria. AB - BACKGROUND: The most appropriate management of penetrating neck injury (PNI) remains controversial. This study was conducted to determine the accuracy and safety of physical examination as the basis of selective observational management of PNI at our institution. METHODS: The study was conducted between 1991 and 2006. Patients whose injuries penetrated platysma were included. Following resuscitation, physical signs were utilised to select patients for exploration or observation. Investigations were based on physical signs which, with details of injured structures, treatments and outcomes, were recorded. RESULTS: There were 225 patients of whom 209 (93.0%) were men. Their mean age was 28 years. The majority (74.2%) of cases were stab wounds, and the balance (25.8%) were gunshot injuries. In 37.8% and 27.6% of patients, injuries were sustained during armed civilian conflicts and robberies, respectively. Patients with no signs of significant injuries (37.8%) were treated by observation. Overall, 52.4% underwent neck exploration; injuries requiring repair were found in 87.3% of these patients. Physical signs as a basis of detecting significant injury had a sensitivity of 97.2% and specificity of 87.4%. Overall mortality was 4.0%. CONCLUSION: Physical examination can accurately select patients with PNI who can be safely managed by observation. Physical signs can also identify patients who require further diagnostic evaluations. PMID- 19813444 TI - Aortocaval fistula--rare complication of ruptured abdominal aortic aneurysms. PMID- 19813445 TI - Ogilvie's syndrome, beta-blockers and burns. PMID- 19813446 TI - Eponymous yet anonymous. PMID- 19813447 TI - Treatment of septicaemia and severe bacterial infections in foals with a new cefquinome formulation: a field study. AB - A multicentre field study was conducted in accordance with VICH Guideline on Good Clinical Practice (VICH 2000) to confirm the efficacy and safety of a new formulation of cefquinome for the treatment of naturally occurring severe bacterial infections and septicaemia in foals. Thirty-nine foals suffering from severe bacterial infections (such as pneumonia, gastro-enteritis, arthritis, omphalitis, or wound infections) or acute septicaemia were treated twice daily with the test product (1 mg cefquinome/kg body weight) intravenously for three days and then intramuscularly for three to 11 days. Investigators examined the foals daily and scored both systemic and local clinical signs to assess the response to treatment, treatment success and relapses. On the day of inclusion a blood sample was taken from each foal for IgG determination and blood culture. In case of abnormal clinical findings additional samples were taken for bacteriology. Treatment was successful in 87.2% of cases (34 of 39 foals) and no relapses were observed. The average duration of treatment was 7.5 days. At inclusion, bacterial culture was positive in 40.5% (15 out of 37) of the blood cultures. Escherichia coli, Clostridium perfringens and Staphylococcus spp. were the most common isolates and were all susceptible to cefquinome. E. coli predominated in swabs from umbilical and open wound infections, and in rectal swabs E. coli. There was no correlation between IgG at inclusion and study outcome or treatment duration. The test product was very well tolerated by all of the foals following intravenous and intramuscular injection. The cefquinome formulation tested was effective and safe in the treatment of severe bacterial infections and septicaemia in foals under field conditions. PMID- 19813448 TI - Expression of heat shock proteins in tissues from young pigs exposed to transport stress. AB - The objective of the present study was to investigate the role of heat shock proteins (Hsps) as potential stress response marker in several organs of transported pigs. Constitutive (Hsp90, Hsp70 and Hsp27) and inducible (Hsp72 and Hsp86) Hsps expressed in skeletal muscle, heart, liver and kidney of transported young pigs were investigated. The study comprised 13 German Landrace line pigs (mean weight, approx. 35 +/- 1 kg). Eight animals (n = 8) were transported for a period of 6 h, whereas the control group (n = 5) was kept under normal housing conditions. Identification of Hsps was performed by immunoblot using porcine specific antibodies. Although the 5 Hsps were regularly detected in all porcine tissues, no clear up-regulation could be observed due to transportation. Densitometrical analysis of the immunoblots revealed an unexpected result. Every tissue had a significant reduction of at least 2 Hsp members: Kidney (Hsp90 and Hsp70), M. longissimus dorsi (Hsp90 and Hsp72) and M. gluteus maximus superficialis (Hsp90 and Hsp86), liver (Hsp90, Hsp86 and Hsp27). The heart was most affected, all Hsps were significantly reduced by 26% to 41% after 6 h of transportation. The regular reduction of the large Hsp90 and Hsp86 in nearly all tissues examined point to a new and critical role these Hsps might have in counteracting short-term stress reactions. The fast reduction of Hsps in fatal organs such as heart and kidney may have a relation to organ failure. Hsp90 level may therefore serve as a potential marker for the stress pigs are suffering during transportation. PMID- 19813449 TI - [Mortality in free living siskins (Spinus spinus Linnaeus, 1758) due to Salmonella typhimurium, phage type DT104 and DT013]. AB - This report deals with an enzootic due to Salmonella Typhimurium in two free living Eurasian siskins (Spinus spinus Linnaeus, 1758). Other birds in the vicinity of the siskins were not affected. Clinical signs consisted of non specific symptoms such as ruffled plumage, apathy and reduced food intake. During necropsy, gross lesions were enlarged livers with focal necrosis, pale spleens, enlarged kidneys, pneumonia and enteritis. Salmonella Typhimurium was isolated from internal organs in pure culture. Using the polymerase chain reaction, the detection of Salmonella according to EN ISO 6579:2002 was confirmed. The detailed characterisation of both isolates in the Federal Institute for Risk Assessment and in the Robert Koch Institute yielded for the first siskin Salmonella Typhimurium, 4, 5, 12: i : 1, 2, LT DT104, BT a and for the second siskin Salmonella Typhimurium, 4,12 i : 1, 2, LT DT013, BT c. These phage types were identified for the first time in siskins. The detected phage types have importance as causes of disease not only for free living siskins but also as infectious and zoonotic agents for domestic poultry and poultry products. PMID- 19813450 TI - A first case of ehrlichiosis in a horse in Poland. AB - The study was aimed at determining the cause of a disease in a horse exhibiting symptoms of fever, joint effusion, weakness, and extravasations on the mucous membranes. Blood was drawn from the animal for haematological and biochemical molecular tests. The PCR technique revealed the presence of 16S RNA Ehrlichia spp. genetic material in the blood samples. DNA amplification by means of primers EHR 521 and EHR 747 gave a product with a volume of 247 bp.The sequence of the PCR product obtained showed a 97.6% similarity with a sequence of a fragment of 16S RNA Ehrlichia phagocytophila, gene number EU 090186 in the GenBank. A microscopic examination of blood smears demonstrated the presence in the neutrophiles of intracytoplasmatic structures characteristic for Ehrlichia.The treatment involved the intravenous application of oxytetracycline at a dose of 8 mg/kg of body mass for 7 days, and resulted in a gradual recovery. The positive results of the molecular test and the positive reaction to the therapy are an indication of the first case of ehrlichiosis in a horse in Poland as confirmed in laboratory test. PMID- 19813451 TI - Immunohistological demonstration of Rhodococcus equi in a trotter foal. AB - A 3-month-old female trotter foal was euthanized due to severe dyspnoea. Pathomorphologically a chronic granulomatous to necrotizing pneumonia was found and Rhodoccocus (R.) equi was isolated microbiologically. An immunohistological method using a murine monoclonal antibody against a 15-17 kDa antigen of virulent R. equi was established in formalin-fixed and paraffin-embedded tissue sections using various antigen retrieval techniques to optimize the staining results. Microwave treatment was most suitable for the demonstration of bacterial antigen localized predominantly in intralesional macrophages. Immunohistology is an additional method for identifying R. equi-infections in equine tissue and may be useful in retrospective studies on paraffin-embedded archive material. PMID- 19813452 TI - Economic downturn hits ORs, but few layoffs of periop staff. PMID- 19813453 TI - RI standardizes safe-site protocol. PMID- 19813454 TI - Fine-tuning tissue management. PMID- 19813455 TI - Artificial intervertebral disc replacement for symptomatic cervical disc disease. PMID- 19813456 TI - Recession tests surgery center staffing. PMID- 19813457 TI - Hospitals to report on normothermia. PMID- 19813458 TI - An engineer's eye on turnover time. PMID- 19813459 TI - Time may be right to buy equipment. PMID- 19813460 TI - Liquid chromatography-mass spectrometry method for the determination of amlodipine in human plasma and its application in a bioequivalence study. AB - A sensitive and selective liquid chromatographic-mass spectrometric (LC-MS) method for the determination of amlodipine (CAS 88150-42-9) in human plasma has been developed. Sample preparation was based on liquid-liquid extraction using NaOH and cyclohexane. Analytical determination was carried out on a C18 column interfaced with a single quadrupole mass spectrometer. Positive electrospray ionization was employed as the ionization source. The mobile phase was 10 mmol/L ammonium acetate solution-methanol (30:70, v/v) at the flow rate of 0.2 mL/min. The method was linear in the concentration range of 0.2-20 ng/mL. The lower limit of quantification was 0.2 ng/mL. Amlodipine was sensitive to UV light. The method was validated in terms of accuracy, precision, absolute recovery, and stability. The intra- and interday relative standard deviation across three validation runs over the entire concentration range was less than 8.17%. The accuracy determined at three concentrations (0.4, 2.0 and 10 ng/mL for amlodipine maleate) was within +/- 3.17% in terms of relative error. The method herein described was successfully applied for the evaluation of pharmacokinetic profiles of amlodipine maleate tablets in 20 healthy volunteers. The results showed that AUC, Tmax, Cmax and T 1/2 between the test and reference formulation have no significant difference (P > 0.05). The relative bioavailability was 103.7 +/- 12.3%. PMID- 19813462 TI - Truncated area under the urinary excretion rate curve in the evaluation of alendronate bioequivalence after a single dose in healthy volunteers. AB - This study was designed to evaluate the bioequivalence of two formulations of alendronate (CAS 121268-17-5) 70 mg (test formulation, alendronate 70 mg tablets, vs. the reference formulation) in 80 healthy volunteers under fasting conditions. The trial followed an open, randomized, crossover design with a washout period of 28 days. Urine samples were collected up to 48 h post-dose, and the concentrations of alendronate were determined by HPLC. The mean Ae(0-48) was (mean +/- SD) 152.15 +/- 136.09 microg for the reference formulation and 150.37 +/- 126.20 microg for the test formulation, while the mean Rmax was 53.33 +/- 41.53 microg/h and 55.85 +/- 49.57 microg/h, respectively. No significant differences in pharmacokinetic parameters between the two formulations were found. The 90% confidence interval for the ratios of Ae(0-48) and Rmax of alendronate were within the acceptance range for bioequivalence trials. The results of the present study suggest that the test formulation is bioequivalent to the reference formulation. The analyses of truncated AURC to shorter times showed similar values, which were within the range of bioequivalence. PMID- 19813461 TI - Bioequivalence study of two tablet formulations of ramipril in healthy volunteers. AB - This study was conducted in order to assess the bioequivalence of a test and reference tablet formulation containing 10 mg of ramipril ((1S,5S,7S)-8-[(2S)-2 [[(1S)-1-ethoxycarbonyl-3-phenyl-propyl]amino]propanoyl]-8-azabicyclo[3.3.0] octane-7-carboxylic acid, CAS 87333-19-5). Forty healthy male and female volunteers were treated in a single-centre randomised, single-dose, open-label, 2 way crossover study, with a washout period of 35 days between treatments. Plasma samples were collected up to 168 h post-dosing for the determination of ramipril and its active metabolite, ramiprilat, by LC-MS/MS. The evaluation of bioequivalence was based on the following pharmacokinetic parameters that were calculated by standard non-compartmental methods: the area under the plasma concentration-time curve from zero to the time of the last quantifiable concentration (AUCt) and that extrapolated to infinity (AUC) and the maximum observed concentration (Cmax). The 90% confidence interval of the ratios (test/reference) (obtained by analysis of variance, ANOVA) were 0.83-1.20 for Cmax of ramipril, 0.90-1.10 for Cmax of ramiprilat, 0.95-1.23 for AUC(0-48) of ramipril, 0.97-1.11 for AUC(0-168) of ramiprilat, 0.96-1.23 for AUC of ramipril and 0.98-1.15 for AUC of ramiprilat, i.e. within the predefined acceptable range for the conclusion of bioequivalence. Tmax of the test formulation was 0.67 +/- 0.33 h for ramipril and 2.28 +/- 0.74 h for ramiprilat; Tmax of the reference formulation was 0.71 +/- 0.32 h for ramipril and 2.40 +/- 0.88 for ramiprilat. The ramipril and ramiprilat Tmax values estimated for the test and the reference formulations were not significantly different (p-value > 0.05). The study indicated that the test and reference formulations containing 10 mg of ramipril were equivalent in terms of both the rate and extent of bioavailability. PMID- 19813463 TI - Efficacy and safety of piroxicam patch versus piroxicam cream in patients with lumbar osteoarthritis. A randomized, placebo-controlled study. AB - In order to assess the efficacy and safety of a new patch containing 14 mg of piroxicam (CAS 36322-90-4) 1%, applied once daily, in comparison with a reference marketed formulation, piroxicam 1% cream applied three times a day, placebo patch applied once daily, a randomized, placebo-controlled, parallel-group clinical trial was carried out by general practitioners in patients with lumbar osteoarthritis aged between 18 and 75 years. Pain during daily activities scored on a 100 mm visual analogue scale was the primary outcome measure. Other secondary outcome measures were pain on isometric contraction, on full passive motion, and on pressure, and functional disability. Statistical analysis was performed on the differences between the three groups in the intention-to-treat population (ITT). One hundred and eighty patients were enrolled. The available ITT population comprised 179 patients. The compliance was very good. Decrease in pain score during daily activities after the eight days of study treatment (at the final visit, Vf) was 42.2%, 41.7% and 25.8% in the piroxicam patch, piroxicam cream and placebo groups, respectively. The difference between the pain scores in two active treatments arms was not statistically significant at the Vf whereas the differences between the pain scores of two active treatment arms vs the placebo arm were statistically significant validating the study design. All efficacy measures improved during the study, for both the active treatment groups, and the results for the secondary efficacy variables were generally consistent with those concerning the main efficacy criterion. The difference between the two active treatments in pain during daily activities were statistically significant at the final visit; in fact the 95% CI of the difference between the mean of responder rate of the piroxicam patch and piroxicam cream was -18.3%, +24.4% indicating a trend of superiority of the piroxicam patch versus the cream (per-protocol analysis). The data obtained during the intermediate visit (V2, day 4) allow us to assess that the piroxicam patch was on average better than the piroxicam cream in terms of fast pain reduction (change from baseline: - 29.1% for piroxicam patch in comparison to 24.6% for piroxicam cream). Moreover the piroxicam patch proved to be on average more effective than the piroxicam cream in terms of secondary efficacy endpoints. Safety was considered satisfactory in all groups. CONCLUSIONS: The piroxicam patch is effective in the treatment of lumbar osteoarthritis and has demonstrated to be well tolerated and it improves patients compliance. The piroxicam patch offers a comparable alternative to the marketed piroxicam cream for the treatment of lumbar osteoarthritis with the advantage of a better compliance with the once a day application of the patch compared to three daily applications for the piroxicam cream. PMID- 19813464 TI - Possible link between history of hypersensitivity to a specific non-steroidal anti-inflammatory drug (NSAID) and positive results following challenge test to alternative NSAIDS. AB - INTRODUCTION: In subjects with hypersensitivity reactions to non-steroidal anti inflammatory drugs (NSAIDs), the choice of suitable alternative drugs with the lowest risk of reaction is imperative for therapeutic management. A safe method to exclude drug hypersensitivity is to perform a challenge test for an alternative drug. The present study was conducted to: obtain more information about the safety of NSAIDs; assess the risk of reaction following the administration of a selective or nonselective cyclooxygenase 2 (COX-2) inhibitor in patients with a history of adverse reactions to NSAIDs; investigate if age and/or gender play a role in the susceptibility to develop adverse reactions to NSAIDs. PATIENTS AND METHODS: This retrospective study includes 524 patients with a history of hypersensitivity to NSAIDs admitted to undergo challenge test to an alternative anti-inflammatory drug. Statistical significance was achieved when odds ratio (OR) and risk ratio (RR) values were >1. RESULTS: 8.39% of patients with hypersensitivity reactions to NSAIDs showed a positive challenge test for the alternative drug. Challenge tests for nonselective COX-2 inhibitors were positive in 16.2% of patients with previous reaction to a same drug class and in 12.9% of patients with a history of reaction to selective COX-2 inhibitors. No positive challenge test to a non-selective COX-2 inhibitor was found in patients with a history of hypersensitivity to nimesulide (CAS 51803-78-2). Challenge tests for selective COX-2 inhibitors were positive in 4.6% of patients with a previous reaction to nonselective COX-2 inhibitors and in 7.2% of patients with a history of reaction to selective COX-2 inhibitors. The RR of a positive challenge test to a non-selective COX-2 inhibitor was significant in patients who had a history of reaction to an analogous compound (P 0.21, OR 1.31, RR 1.26). DISCUSSION: In this study, selective COX-2 inhibitors represented the class of NSAIDs less frequently reported as responsible of adverse reaction. These data underline that there is a higher risk to find a positive challenge test to a non selective COX-2 inhibitor than to a selective one in patients with previous adverse reactions to a non-selective COX-2 inhibitor. Moreover, the data evidence that females could have a higher risk compared to males to develop an adverse reaction to selective COX-2 inhibitors. In conclusion, it appears necessary to pay attention to the kind of NSAIDs reported as the cause of hypersensitivity in anamnesis, because it must be considered a successful guide in choosing the alternative drug to administer to the patient during the challenge test. PMID- 19813465 TI - Synthesis of novel tadalafil analogues and their evaluation as phosphodiesterase inhibitors and anticancer agents. AB - Two closely related series of novel beta-carboline derivatives, electronically similar to tadalafil (CAS 171596-29-5), were synthesized and evaluated for their inhibitory effects upon phosphodiesterase 5 (PDE5) and phosphodiesterase 11 (PDE11) and their in vitro tumor cell growth inhibitory activity versus HT29 colorectal carcinoma cell line. Interestingly, some of the synthesized compounds showed growth inhibitory properties that appear to be associated with their ability to inhibit PDE5. Moreover, the PDE5 inhibition seems relevant to the stereochemical aspects of the compounds. PMID- 19813466 TI - Chromatographic/mass spectrometric method for the estimation of itraconazole and its metabolite in human plasma. Application to a bioequivalence study. AB - A HPLC/mass spectrometry method for the estimation of itraconazole (CAS 84625-61 6, ITR) and its active metabolite hydroxyitraconazole (CAS 112559-91-8, HOX) in human plasma was developed. Terconazole (CAS 67915-31-5) was used as an internal standard. The analytical method was fully validated according to FDA and EMEA requirements. The accuracy and precision of the developed method was satisfactory and stability studies showed an acceptable variation (below 15%) of ITR and HOX concentrations when the samples were stored frozen at -75 degrees C for 95 days. The developed method was successfully used for a comparative 2 x 2 period, crossover bioequivalence study of two preparations of ITR (Itrakonazol Genexo 100 mg as the test drug) performed on 36 healthy volunteers. PMID- 19813467 TI - Pharmacokinetics and comparative bioavailability study of two clarithromycin suspensions following administration of a single oral dose to healthy volunteers. AB - The aim of the present study was to compare the pharmacokinetics and bioavailability of two commercial brands of clarithromycin (CAS 81103-11-9) suspensions in healthy male Iranian volunteers. In an open label, single-dose, randomized study with a crossover design an equivalent 500-mg clarithromycin suspension was given orally to each of 24 subjects as a single dose on two treatment days. The treatment periods were separated by a one-week washout period. Blood samples were drawn at different time points and the separated plasma was kept frozen at -20 degrees C for subsequent analysis. The plasma concentrations of the drug were analyzed by a rapid and sensitive HPLC method with UV detection. Mean maximum serum concentrations of 2256.5 +/- 590.1 ng/mL and 2840.2 +/- 717.5 ng/mL were obtained for the test and reference formulation, respectively. The AUC(0-infinity) of clarithromycin was on average 45008.7 +/- 10989.9 ng x h/mL for the test and 45221.3 +/- 2155.7 ng x h/mL for the reference formulation. The calculated 90% confidence intervals for the ratio of Cmax (81.98 94.26%), AUC(0)(t) (91.6-109.15%) and AUC(0)(infinity) (93.08-110.85%) values for the test and reference products were all within the 85-120% interval proposed by the FDA and EMEA. Therefore the clarithromycin suspension of the test and reference formulations are bioequivalent in terms of rate and extent of absorption. PMID- 19813468 TI - Vibrant Soundbridge middle ear implant in mixed hearing loss. Indications, techniques, results. AB - OBJECTIVES: The aim of this article is to illustrate the aetiologies of mixed hearing loss that can benefit from a Vibrant Soundbridge (VSB) middle ear implant, the techniques performed and the first results. MATERIALS AND METHOD: The authors report their experience of 13 implantations in mixed hearing loss due to otosclerosis, sequelae of chronic otitis media and congenital aural atresia. The VSB implant was implanted alone or in association with another middle ear surgical procedure, on the ossicular chain or on the round window membrane. RESULTS: The average auditory gain for all patients is 32 dB for pure tone thresholds, and 25 dB for speech recognition. It results from the addition of a gain on the conductive hearing loss by direct stimulation of the inner ear, to a gain on the sensorineural hearing loss by amplification. CONCLUSION: Middle ear implants are the only hearing aids affording a gain in both the conductive and sensorineural components of mixed hearing losses. PMID- 19813469 TI - Vibrant Soundbridge for hearing restoration after chronic ear surgery. AB - INTRODUCTION: Middle ear surgery is primarily concerned with resolving the discharging pathology, in the case of chronic otitis media (COM), or with complete eradication, in case of cholesteatoma. Either of these procedures may require repeated surgeries, often resulting in severe mixed hearing impairment. A middle ear implant may be indicated in these cases instead of a hearing aid because the anatomical conditions in such cases often impede an adequate acoustic coupling. The objective of this study was to evaluate MED-EL Vibrant Soundbridge (VSB) implantation in patients with severe conductive and mixed hearing loss occurring after middle ear surgery for cholesteatoma or chronic otitis media (COM). MATERIALS AND METHODS: Over a 2-years period, the VSB system was implanted in 40 patients between 35 and 81 year old (mean: 59.5). Surgery was performed with comparable technique in 3 regional hospitals in Italy: Rovereto (n=16), Meran (n=12) and Tortona (n=12). The 40 candidates for implantation had a history of 1-5 previous surgeries. Of those, 20 patients suffered from COM and 20 from, cholesteatomas. The floating mass transducer (FMT) of the VSB was placed and stabilized on the round window niche in 32 cases; alternative positioning was necessary in 8 cases. Bone conduction (BC) was tested 1 day post-operatively. At 1 month post-surgery and between 6-9 months, open-field warble tones threshold in VSB-off and VSB-on conditions and open-field speech audiometry for words in quiet were conducted. RESULTS: Results of BC audiometry one day post-operatively showed no significant changes in hearing. Unaided mean pure tone average (PTA4) was 82.38 dB SPL with a mean speech recognition threshold (SRT) of 94.28 dB SPL. Results obtained after a minimum of three months post-operatively were evaluated in terms of aided thresholds and functional gain. At VSB activation, the mean PTA4 was 50.63 dB SPL with a mean SRT of 61.68 dB. After 6-9 months, the group had a mean PTA4 of 47.89 dB SPL and a mean SRT of 53.33 dB SPL. CONCLUSIONS: Implantation of the VSB with its direct driver of the inner ear fluids appears promising for auditory rehabilitation of severe mixed hearing loss associated with sequelae of cholesteatoma surgery. Patients' results improved over time, allowing us to assume a positive effect of consolidation of the coupling related to fibrosis. Results reported here refer to 6-9 months of observation and do not provide evidence of long term stability. PMID- 19813470 TI - [Side effects and patients expectations after vestibular tests]. AB - OBJECTIVE: Patients satisfaction and side effects assessment after vestibular tests. MATERIALS AND METHODS: Questionnaires were sent to 150 patients after vestibular testing in 2006. Forty-two questionnaires were sent back to us. On each questionnaire there were 5 questions regarding patient expectations, 4 questions about side effects after vestibular tests and one question was related to follow-up medical attention. RESULTS: Most of the patients think that the delay between clinical consultation and vestibular test was not too long. Information patients received beforehand regarding vestibular tests aim, duration, possible side effects and diagnostic expectations are good enough for most of the patients. However patient's assessment of the informed consent document and location of vestibular tests is diverse. The vast majority of patients feel "very important" to receive information about vestibular tests results on the same day. For most of the subjects caloric test is the most disconfortable tests. In the following days, more than half of the subjects feel vestibular symptoms. The most frequent suggestion we received from patients is to give them a phone calls after vestibular tests. CONCLUSION: It is worth mentioning the importance of informing the patient about vestibular test results upon completion. Our aim is to make some changes on the informed consent document given to patients and to make a phone call to patients after vestibular tests. PMID- 19813471 TI - [Carcinogenesis of the ethmoidal adenocarcinoma due to wood dust]. AB - OBJECTIVES: To recognize the mechanisms and the different oncogenic pathways of ethmoid adenocarcinoma (EADC) in woodworkers. METHODS: A systematic review of the literature and evaluation according to scientific evidences. RESULTS: Wood dust less than 10 microm settles on the external aspects of the ethmoid sinus that is the middle turbinate, the middle meatus and the olfactory cleft. The risk of developing an EADC is important from the first year. It is impossible to know what determines the latency period. Usually, this period exceeds 30 years and only 10% of patients are younger than 50 years of age. Chromosomal instability of the respiratory mucosa appears to be one of the major phases in the oncogenic process. Certain biomolecular and genetic factors are shared with the adenocarcinoma of the colon but they are not activated with the same importance and in the same context suggesting two distinct mechanisms of evolution. The local anatomo-clinical setting in the nasal cavity and carcinogenic substances in the wood dust play a key specific role in the development of EADC. No oncoproteins or immunohistochemistry features were identified in the process suggesting a cascade of genetic or molecular series of events. PMID- 19813472 TI - Moderate leukocyte infiltration in the lower turbinate mucosa after a two-week intranasal administration of mometasone furoate, azelastine or salmon calcitonin. AB - OBJECTIVE: We used light microscopy to search for local changes of the nasal mucosa associated with daily intranasal administration of mometasone furoate (MF), azelastine (AZ) or salmon calcitonin (SC). STUDY DESIGN: Biopsies of the lower nasal turbinate were obtained from four groups of 8 individuals after 14 days of daily administration of a saline solution (control group) or of MF AZ and SC. METHODS: Small biopsies of the anterior portion of the lower nasal turbinate were collected with the help of a Hartmann forceps under direct visual inspection. The samples were processed for light microscopy and morphometric analysis. Inflammatory infiltration (neutrophils and lymphocytes) of the nasal mucosa was evaluated by a semiquantitative method. Unpaired t test and Bernoulli distribution were applied to evaluate statistical differences between data from the different groups of samples. RESULTS: Samples of the turbinate mucosa of all of the drug-treated groups showed a moderate enhancement in infiltrating neutrophils when compared with the samples from the control group. Infiltration of lymphocytes in the turbinate chorion was significantly different from controls only in the MF and AZ-treated groups. CONCLUSIONS: Intranasal treatment with MF AZ or SC does not cause significant changes in the general architecture of the nasal mucosa. A moderate inflammatory response of the turbinate mucosa, that was expressed by leukocyte infiltration of epithelium and chorion, was observed in all of the 3 drug-treated groups of patients after the 2-week course of intranasal deposition of MF, AZ or SC. PMID- 19813473 TI - [Bronchoscopic findings in children with prolonged respiratory symptoms]. AB - OBJECTIVE: This retrospective study aims to establish an algorithm indicating a bronchoscopy, based on clinical and radiological criteria predictive of the presence of a foreign body (FB), in children with prolonged respiratory symptoms and no history of foreign body aspiration (FBA); to establish a study of these criteria and to compare the clinical, radiological and broncoscopic findings in such cases. METHODS: We chose to review the records of 73 children (age < or = 15 years) with prolonged respiratory symptoms (for at least 15 days) and no history of FBA who underwent bronchoscopy at our institution between 1996 and 2005. RESULTS: The mean age was 3 years and 2 months, the majority of the patients were between 1 and 3 years of age (56%), 59% of the patients were boys and the mean of evolution of symptoms before the broncoscopy was 3 months and 3 weeks. We found a foreign body (FB) in 17 cases (23.2%). The FBs were in 88.2% of the cases of vegetable origin. A granulation tissue was associated in 47% of positive bronchoscopies. There were no complications related to bronchoscopy. The clinical and the radiological findings were no specific of FBs. CONCLUSION: Bronchoscopy must be indicated in suspected cases of FBA to prevent delayed diagnosis and pulmonary sequelae. The clinical and radiological findings may help the clinician to indicate it. PMID- 19813474 TI - [Objective tinnitus and essential palatal tremor in children: report of a case]. AB - Palatal tremor is a rare neurotological disorder responsible for objective tinnitus in children. Palatal tremor may be symptomatic of an underlying neurological disease or essential when a cause cannot be identified. We report a case of an essential palatal tremor in a 10-year-old girl complaining of clicking tinnitus. No treatment was undergone as she was not obviously bothered by the ear clicking sound. Different treatment modalities have been used for distressing tinnitus related to palatal myoclonus. Recently several publications reported satisfactory results with botulinum toxin injection, which seems to be the treatment of choice. PMID- 19813475 TI - Orbital apex syndrome following inferior turbinate radiofrequency. AB - INTRODUCTION: We report a case of orbital apex syndrome following turbinate radiofrequency. METHODS: The clinical features, investigations (nasofibroscopy, supra-aortic and trans-cranial vessels ultrasounds, CT- and MRI-scans) are described. RESULTS: A forty-year-old man underwent radiofrequency volumetric tissue reduction (RFVTR) for bilateral inferior turbinate hypertrophy. No particular problems were reported during the procedure under general anaesthesia. Immediately after the general anaesthesia, the patient complained of right eyelid ptosis with right monocular blindness. The patient also had ophthalmoplegia and suffered from right corneal anaesthesia and right hypoaesthesia of the cheek. The CT-scan showed right ethmoidal and maxillary sinusitis with no bone or tissue lesions. MRI-scan showed an enlarged aspect of the subarachnoid membrane of the right optic nerve. Corticosteroid treatment was prescribed but did not produce any satisfactory result. No improvement in visual acuity was observed. CONCLUSION: Turbinate radiofrequency may result in a definitive orbital apex syndrome. PMID- 19813476 TI - [Transverse maxillary deficiency: interest in the management of chronic nasal obstruction]. AB - OBJECTIVES: To describe the surgical orthodontic technique of correcting transverse maxillary deficiency associated with nasal obstruction and malocclusion. SURGICAL PROCEDURE: The surgical-orthodontic correction of transverse maxillary deficiency consists in an incomplete Le Fort I procedure, without down-fracture, associated with a sagittal palatal osteotomy. This procedure is called SARE (Surgically Assisted Rapid Expansion). A pre-fabricated orthodontic appliance is cemented to premolars and molars prior to the surgical procedure. The patient activates the appliance allowing a 1 mm widening a day up to 10 mm. A stabilisation phase is mandatory to avoid early relapse of the correction. This phase involves ossification of the midline osteotomy site. RESULTS: A 22 year old female consulted for a chronic nasal obstruction. Previous treatments, medical and surgical (septoplasty) did not improve her symptoms. She presented with a typical transverse maxillary hypoplasia. She underwent a surgery with the technique described above. Nasal patency improvement was noted on the first post-operative day. This was confirmed by comparing pre and post-operative rhinomanometry. An orthodontic treatment followed for several months to stabilize the result. DISCUSSION AND CONCLUSION: Main advantages of this surgical intervention are to correct both the dental malocclusion and the nasal obstruction by widening the nasal floor and the maxillary arch. Post-operative period is mostly uneventful. It can take up to six months to achieve a good result that is why a strong collaboration has to exist between the surgeon, the orthodontist and the patient. PMID- 19813477 TI - [Atypical laryngo-tracheal post-intubation complications: three case reports]. AB - Endotracheal intubation can induce iatrogenic laryngeal and tracheal complications, particularly when the intubation lasts for a long time. The ENT pratician's job is not only to cure the laryngeal sequelaes but also to prevent and diagnose them. Authors report three atypical cases of laryngeal and tracheal post-intubation complications (laryngeal bilateral immobility, interarythenoidal synechia, oesotracheal fistulae) and discuss them regarding medical literature data. PMID- 19813478 TI - [Laryngeal sarcoidosis: case report]. AB - Sarcoidosis is a chronic disease characterized by non-caseating granulomatous inflammation in one or multiple organs. The most commonly involved sites are the lungs and the skin. Involvement of the larynx is rare but significant. We present here a case of biopsy-proven of laryngeal sarcoidosis with favourable evolution without specific treatment. PMID- 19813479 TI - Courage and character, leaders and legends: an interview with Maria Siemionow, MD, PhD, DSc. PMID- 19813480 TI - Quality initiatives in transplantation. PMID- 19813481 TI - Use of failure mode and effects analysis for proactive identification of communication and handoff failures from organ procurement to transplantation. AB - A multidisciplinary team from the University of Wisconsin Hospital and Clinics transplant program used failure mode and effects analysis to proactively examine opportunities for communication and handoff failures across the continuum of care from organ procurement to transplantation. The team performed a modified failure mode and effects analysis that isolated the multiple linked, serial, and complex information exchanges occurring during the transplantation of one solid organ. Failure mode and effects analysis proved effective for engaging a diverse group of persons who had an investment in the outcome in analysis and discussion of opportunities to improve the system's resilience for avoiding errors during a time-pressured and complex process. PMID- 19813482 TI - A system's approach to improve organ donation. AB - Using lessons learned from the US Department of Health and Human Services National Donation Breakthrough Collaborative, New York-Presbyterian Healthcare System (NYPHS) partnered with 5 donor service areas covering its member hospitals to improve donation across the system. By integrating established communication networks with the "spread" techniques of the Breakthrough Collaborative, the NYPHS identified hospital champions and best practices and established standardized outcome metrics. The improvements that resulted were a sustained increase of 40.23% in consent rate and an initial 41.7% increase in conversion rate during the first 6 months, although that conversion rate was not sustainable. During the 8 measured periods, 21 hospitals met or exceeded the 75% conversion rate during 1 or more quarters. NYPHS was able to spread these successes and outcome metrics through its established communication networks of quarterly report cards, regular senior leader meetings, and real-time access to a secure member-only Web site, thus keeping organ and tissue donation at the forefront of hospital leaders' priorities. PMID- 19813483 TI - Evolution of quality at the Organ Center of the Organ Procurement and Transplantation Network/United Network for Organ Sharing. AB - One of the goals of the Organ Center of the Organ Procurement and Transplantation Network/United Network for Organ Sharing is to increase the efficiency of equitable organ allocation in the United States. Recognizing the ever-growing need for organ donors and transplants, leaders at the Organ Center increased its commitment to quality improvement initiatives through the development of a quality management team in 2001. The Organ Center began to focus on ways to capture data on processes and pinpoint areas for improvement. As the collection and analysis of data evolved, the Organ Center embraced formal quality standards, such as improvement cycles. Using these cycles, the Organ Center has seen significant improvement. One initiative involving lifesaving heart, lung, and liver placement showed success by doubling the Organ Center's organ placement rate. Another project involving the validation of donor information demonstrated that the accuracy of organ allocation can be improved by 5% on a consistent basis. As stewards for the gift of life and leaders in organ allocation, the Organ Center uses continuous quality improvement to achieve the goal of increasing the efficiency of equitable organ allocation. PMID- 19813484 TI - Attaining specific donor management goals increases number of organs transplanted per donor: a quality improvement project. AB - Most organ procurement organization professionals and transplant surgeons intuitively know that meeting donor management goals improves organ allocation and transplant outcomes. In this era of evidence-based medicine, it is important to know whether the data support this assumption. All 6 organ procurement organizations in the United Network for Organ Sharing's region 10 agreed on 6 specific donor management goals. The organ procurement organizations then compared the number of organs transplanted per donor when goals were met with the number when goals were not met. Results were broken down by donor type: standard criteria donation, expanded-criteria donation, and donation after cardiac death. For all 6 organ procurement organizations combined, the data for all of 2008 show a substantial and statistically significant improvement in number of organs transplanted per donor for standard criteria donation and total donors when goals are met, with a smaller degree of improvement (although not statistically significant) in the number of organs transplanted per donor for expanded-criteria donation and donation after cardiac death when goals are met. PMID- 19813485 TI - Lung donation after cardiac death of recent heart transplant recipient. AB - In June 2008, Gift of Life Michigan was notified of a potential donor. After the initial chart review, it was determined that the patient would be a candidate for donation after cardiac death. Two things made this case interesting: (1) the patient was an organ recipient himself, having received a heart transplant just a short time before his death, and (2) even though this was a donation after cardiac death and the patient had recently received the heart transplant, the patient's lungs and the left kidney were successfully recovered and used for transplant. PMID- 19813486 TI - Engaging the third estate: the Transplant Growth and Management Collaborative. AB - The Organ Donation and Transplantation Collaboratives that occurred within the United States from 2004 to 2008 helped contribute to a significant increase in organ donors and transplants across the country. Centers were needed to accommodate and maintain this increase in capacity to perform successful transplantations for candidates on the waiting list. The Transplant Growth and Management Collaborative was created to help fulfill this new performance level expectation. In 2007 the US Department of Health and Human Services, Health Resources and Services Administration published a best-practice report based on high-performing centers that experienced a significant increase in volume while maintaining expected, or higher than expected, outcomes. The report produced a change package that outlined common strategies, key change concepts, and actions used at the best-practice centers that could be adapted by other transplant programs by using Plan-Do-Study-Act cycles to test the impact of the changes. This change package and use of the Plan-Do-Study-Act cycles formed the foundation of the Collaborative that occurred from October 2007 through October 2008 to spread best practices to transplant programs willing to commit to making changes that could result in a 20% increase in transplant volume. More than 120 transplant centers participated at some point in the Collaborative. Although preliminary results of the Collaborative show that only a few participating programs achieved the 20% volume increase goal, many participating centers reported putting successful models in place for each of the strategies identified in the best-practice change package. PMID- 19813489 TI - Donor factors that affect the number of organs transplanted per donor. AB - BACKGROUND: Demographic factors and factors from donors' medical and social history influence the number of organs transplanted per donor. The goal for organ procurement organizations is 4.30 organs transplanted per standard criteria donor. OBJECTIVE: To examine the influence of factors related to donors' demographics and donors' medical and social histories on the number of organs transplanted per donor. METHODS: The medical and social histories of 772 deceased donors were reviewed to assess demographic factors (eg, age, sex), diseases (eg, heart disease, hepatitis, diabetes, hypertension), and other factors (eg, body mass index, high-risk behavior, alcohol abuse). Statistics were calculated by using MarketSight software. RESULTS: Between 2003 and 2007, 2409 organs were recovered and transplanted from 772 donors (number of organs per donor, mean [SD], 3.12 [1.73]) in our designated service area. Some demographic factors (eg, age, sex) were associated with significantly fewer organs transplanted per donor. Other factors from donors' medical and social histories also played a major role, with each added factor significantly decreasing the number of organs transplanted per donor. The number of organs transplanted differed significantly between donors with 0 vs 1, 1 vs 2, and 2 vs 3 or more factors related to their medical and social histories. CONCLUSION: Demographic factors and other factors in donors' medical and social histories significantly reduced the number of organs transplanted per donor. An algorithm that uses all potential demographic factors and factors related to medical and social history nested within each other could be developed to predict number of organs transplanted. PMID- 19813487 TI - Correlates of support for living donation among African American adults. AB - CONTEXT: Living donation is studied with much less intensity among African Americans than among the general population. Examination of barriers to living donation can lead to effective strategies to educate dialysis patients and their families about this alternative. OBJECTIVE: To explore the correlates of likelihood of becoming a living donor among community-recruited African American adults. DESIGN/PARTICIPANTS: Cross-sectional data were gathered via self administered questionnaire from 425 African American adults, age 18 years and older, who were recruited from 9 churches in Atlanta. MAIN OUTCOME MEASURES: Self reported likelihood of becoming a living donor to a close family member, an extended family member or friend, or a stranger. RESULTS: More than three quarters of participants were willing to act as living donors to a close family member or spouse and two-thirds to friends or extended family. For likelihood of donating to a friend or extended family member, only willingness to engage in deceased donation was significantly associated; to a stranger, both willingness to engage in deceased donation and attitudes toward donation were significantly associated. Knowledge of and personal experiences with donation and/or transplantation were not significantly associated with likelihood of any type of living donation. CONCLUSIONS: Findings indicate widespread support for living donation to a close family member or spouse. These findings have important implications for dialysis patients who must decide whether to approach friends and/or family about the possibility of serving as a living donor and emphasize the need for interventions to help facilitate this process. PMID- 19813488 TI - Defining high risk in adult kidney transplantation. AB - BACKGROUND: Because identifiable factors contribute to allograft loss, and because no consensus has been reached on the definition of high risk, an interdisciplinary group of nurses, physicians, pharmacists, and social workers was convened in May 2008. OBJECTIVE: Participants sought to reach consensus about the current state of science and best practices related to the definition and management of high-risk kidney transplant recipients. METHODS: An expert facilitator with extensive experience in leading consensus teams guided consensus building activities, which included discussion and small-group work. RESULTS: This consensus group conceptualized the definition of the "high-risk" kidney transplant recipient and provided information to guide the multidisciplinary team in their assessment of these patients before and after transplant. Three key areas, which were conceptualized as independent scales, had a substantial impact on outcomes: (1) transplant recipient medical factors, (2) donor and recipient immunological factors, and (3) transplant recipient psychosocial factors. Though depicted separately, alteration of a specific risk on one scale could influence some risk factors on another scale. In addition, the kidney allograft itself must be considered in the assessment of high risk. CONCLUSIONS: The continuum of risk described here should be useful to transplant clinicians in their assessment of high-risk adult kidney transplant patients, may aid centers in developing a more complete definition of high risk, and may lead to risk-reduction efforts. PMID- 19813490 TI - Effects of administration of intravenous naloxone on gas exchange in brain-dead lung donors. AB - OBJECTIVE: To observe the effect of naloxone on the lung function of potential lung transplant donors with neurogenic pulmonary edema. DESIGN AND INTERVENTIONS: Donors aged 16 to 55 years without any factors to contraindicate lung donation (pneumonia, pulmonary contusion, etc) were included. Ventilator settings were standardized to a tidal volume of 10 to 12 mL/kg, an FIO2 of 0.40, and a respiratory rate that kept PCO2 between 35 and 45 mm Hg. Chest physiotherapy, nebulizer treatments, and frequent suctioning were undertaken. Baseline arterial blood gas analysis and an oxygen challenge were performed. The patients were then given 8 to 10 mg of naloxone. Oxygen challenges and arterial blood gas analyses were repeated every 4 to 6 hours. The data were analyzed by using a paired t test, and each patient served as his or her own control. SETTING: These interventions were performed on the 19 LifeQuest donors who met the set criteria from July 2002 to July 2004. RESULTS: The PaO2 on the oxygen challenge immediately after administration of naloxone increased from 329 (SD 177) to 363 (SD 191) mm Hg, although the increase from baseline was not significant. The PaO2 from the second oxygen challenge (median time, 7 hours after administration of naloxone) increased to 413 (SD 177) mm Hg (P<.01). PMID- 19813491 TI - Development and validation of a tool for assessing understanding of brain death. AB - BACKGROUND: Death by neurological criteria is often misunderstood by laypersons even though they make decisions about withdrawal of care and organ donation. No validated questionnaire for determining laypersons' understanding of brain death exists. Such a tool could be useful in clinical, educational, and research settings. METHODS: Brain death experts and a focus group of laypersons were used to develop a 5-item questionnaire with face validity. The questionnaire explores 3 concepts: apnea, irreversibility, and differentiation between cardiac death, brain death, and persistent vegetative state. The questionnaire was administered to separate groups of laypersons and experts and was readministered 7 to 10 days later. Test-retest reliability for individual items and overall score was measured by using Spearman rank correlation. Internal consistency of the questionnaire was measured by using Cronbach alpha. Utility of the questionnaire in discriminating between scores of laypersons and experts was evaluated by using a t test. RESULTS: Twelve laypersons and 13 experts participated. The test-retest correlation was significant for all questions (Spearman p range, 0.43-0.94) and raw score (Spearman p=0.91, P<.001). Internal consistency was fair (Cronbach alpha=0.64). The questionnaire enabled discrimination of laypersons from experts, with mean (SD) raw scores of 3.0 (1.1) vs 4.8 (0.6), respectively (t test, P<.001). Removal of 1 item improved internal consistency (Cronbach alpha=0.70), but with a corresponding decrease in discriminatory ability. CONCLUSIONS: This simple 5-item questionnaire for evaluating understanding of brain death has test retest reliability, internal consistency, and can be used to discriminate between persons who do and do not understand brain death. PMID- 19813492 TI - Tacrolimus: review of pharmacokinetics, pharmacodynamics, and pharmacogenetics to facilitate practitioners' understanding and offer strategies for educating patients and promoting adherence. AB - Kidney transplantation requires lifelong immunosuppression with agents that prevent allograft rejection. Immunosuppressive regimens typically include a steroid, an immune modulator (eg, azathioprine, mycophenolate mofetil, or mycophenolate sodium), and a calcineurin inhibitor, either cyclosporine or tacrolimus. Tacrolimus is metabolized by cytochrome P450 3A4 in both the liver and small intestine. Drugs that are substrates of cytochrome P450 3A4, as well as inhibitors and inducers of cytochrome P450 3A4, can cause significant interactions with tacrolimus. A review of the pharmacodynamics and pharmacokinetics of tacrolimus is important to enhance practitioners' understanding when using tacrolimus after kidney transplantation. It is also important to educate patients and their families about tacrolimus. Patients' adherence to this medication regimen is pivotal for allograft survival. A consistent and comprehensive approach to education and discharge teaching is a key component of adherence and the attainment of therapeutic drug levels. At Shands Jacksonville Transplant Center, discharge education and teaching tools aid the transplant professionals and facilitate patients' adherence. This in turn supports the goals of maintaining therapeutic serum levels of tacrolimus and improving renal allograft survival. PMID- 19813493 TI - Interview with J. Michael Henderson, MD, Chief Quality Officer, The Cleveland Clinic Health System. Interview by Art Thomson, MA. PMID- 19813494 TI - Resolution 8 is not the answer. PMID- 19813495 TI - The whooping cranes of medicine. PMID- 19813496 TI - Show health the money. PMID- 19813497 TI - Consequences of uninsurance in Wisconsin. AB - BACKGROUND: More than 9% of Wisconsin residents are without health insurance. Several states have passed legislation to increase access to health care and move toward universal coverage. Wisconsin legislators are also considering health care reform proposals that present alternatives to the current system. OBJECTIVE: To describe experiences and concerns of individuals who are uninsured as compared to individuals with health insurance. The purpose is to contribute to the current policy debate on health care reform in Wisconsin. METHODS: Participants in this qualitative study were selected from a set of individuals in west-central Wisconsin who completed a health care survey (n = 222) and gave consent to be contacted for an interview. The interviews were conducted in the spring of 2008; the sample (n = 18) includes equal numbers of individuals with and without health insurance. RESULTS: Regardless of insurance status, participants in this study reported worries about affordability, access to health care, and the influence that health insurance coverage has on their choices in life. Participants agreed that all citizens have a right to health care and favor health care reform to make that right a reality. CONCLUSION: Wisconsin residents, both insured and uninsured, experience worries and uncertainties about health care access and affordability. These qualitative findings provide evidence of public support for health care reform. PMID- 19813498 TI - Wisconsin physician opinions on health care reform. AB - OBJECTIVE: To determine Wisconsin physicians' opinions regarding health care reform. METHODS: The University of Wisconsin Survey Research Center performed a 46-question mail survey of 2500 randomly selected physicians from the Wisconsin Medical Society master list of practicing physicians. Respondents rated opinions on a 5-point Likert scale. Demographics of respondents (sex practice type, geographic location, age) were compared to non-responders and the overall Wisconsin physician population. Data analysis quantified opinions regarding the health care system in Wisconsin and nationally, elements of health care reform proposals, and the role of public policy and government in health care. The analysis emphasized a comparison of primary care versus specialist physician responses. RESULTS: The survey yielded a 38% response rate. Respondent demographics were representative of Wisconsin physicians and very similar to nonresponders. Respondents revealed support for several topics, regardless of the respondent's practice type. Respondents also were in agreement on which elements of reform were most frequently favored and most frequently opposed. Nevertheless, there were many areas where primary care physicians strongly differed from specialists, such as favoring legislation for national health insurance (65.6% primary care versus 46.2% specialist). CONCLUSIONS: Wisconsin physicians responding to this survey expressed dissatisfaction with the health care system in which they currently practice and noted a clear desire for system reform. While most respondents agree on a few key priorities, primary care physicians significantly differ in their preferred strategies for reform and, in particular, the role of government in a reformed system. These results indicate a need for more dialogue and education among physicians in order to achieve a consensus that might help promote reform. PMID- 19813499 TI - Clinical outcomes in a community-based single operator coronary interventional program. AB - BACKGROUND: Physicians who perform fewer than 75 coronary interventional procedures annually and centers where fewer that 200 procedures are performed annually are considered to be low-volume and likely to have higher procedural risk and suboptimal outcomes. METHODS: This was assessed in a retrospective analysis of clinical outcomes in 559 patients who underwent multi-device coronary interventions from 1994 through 2002 (prior to and during the bare metal stent era) performed by a single operator in a community hospital setting with on-site cardiac surgical services. RESULTS: ST- and non-ST-elevation myocardial infarction was present in 36.5% and 10.2% of the population, respectively, with 12.3% of ST-elevation myocardial infarction patients presenting with acute cardiogenic shock. Mortality in all patient subsets was less than 1% when patients with ST-elevation myocardial infarction and cardiogenic shock were excluded. The 6-month target lesion revascularization rate for all devices and in all patient subsets during the study period was 9.1%, and for those treated with coronary stents, the rate was 7.8%. From 1999 through 2002, the rates were 2.1% and 1.9%, respectively. CONCLUSIONS: A low-volume coronary interventional program can achieve angiographic percutaneous coronary interventional success rates and major adverse cardiac events and the need for target lesion revascularization comparable to those in high-volume major cardiac referral centers. These results were obtained even when high-risk patients with ST-elevation myocardial infarction were not excluded from the analysis. PMID- 19813500 TI - Career satisfaction and retention risk among Wisconsin internists. AB - BACKGROUND: Physician career satisfaction has been studied extensively, but career satisfaction as it relates to retention is less well studied. The objective was to assess the relationship between career satisfaction and retention in primary care internal medicine physicians in Wisconsin. METHODS: In this descriptive quantitative study, survey data was assessed for correlations between career satisfaction, risk to retention, and demographics. The survey included 1231 primary care internal medicine physicians in the Wisconsin Medical Directory (2007). MEASUREMENTS: Responses were measured by career satisfaction variables, and demographics and retention variables for the purpose of correlations and regression analysis. RESULTS: Survey responses included 573 physicians. An additional 85 physicians were disqualified. The final survey group included 1146 physicians for a response rate of 50%. A total of 116 physicians (20.2%) reported anticipating leaving their current position, 84 (14.7%) physicians reported anticipating leaving the career of medicine. Identified at risk for retention were 144 (25.1%) physicians. The lowest career satisfaction scores were reported in the areas of compensation (3.19) and practice (3.42) on a scale of 1 (very dissatisfied) to 5 (very satisfied). The highest correlations to retention were associated with practice, followed by compensation satisfaction. The level of significance for this study was identified as 0.05, and the P-value was 0.000. CONCLUSIONS: The study findings reveal a significant risk to the stability of continuity of care for patients, and may cost Wisconsin health care organizations more than $35 million in recruitment costs to replace departing physicians. PMID- 19813501 TI - Painless blood testing to prevent neonatal sepsis. AB - The current US guidelines advise that all women colonized with Group B streptococcus (GBS) at 35-37 weeks, as well as those laboring before this time and all women with GBS urinary tract infections, should be offered intrapartum antibiotic prophylaxis, usually in the form of high-dose intravenous penicillin or ampicillin, unless delivered by planned cesarean section before the onset of labor in a woman with intact membranes. In term and preterm babies who are born to treated women, in addition to babies who act ill, the recommendation is to treat the baby with antibiotics. In certain circumstances, such as when the mother receives an intrapartum antibiotic < 4 hours prior to delivery, the baby receives antibiotics even if the baby appears well. This paper proposes a new process for testing for GBS that involves using the umbilical cord. If this process were used, babies would not need to have blood drawn and would experience less pain. PMID- 19813502 TI - Treatment of intractable abdominal pain patient with Bannayan-Riley-Ruvalcaba syndrome using spinal cord stimulation. AB - OBJECTIVE: This case report presents an application of spinal cord stimulation to a patient with intractable abdominal pain Bannayan-Riley-Ruvalcaba syndrome, that conventional treatment failed to ameliorate. MEASUREMENTS: The patient underwent an uneventful spinal cord stimulator (SCS) trial with percutaneous placement of 2 temporal 8-electrode epidural leads (Medtronic Inc, Minneapolis, Minnesota) to level T6-T7. RESULTS: After experiencing excellent pain relief over the next 3 days, the patient was implanted with permanent leads and a rechargeable generator 4 weeks later and reported sustained pain relief. CONCLUSION: Preliminary outcomes from this case suggest that spinal cord stimulation offers an alternative treatment option for select patients with intractable abdominal pain and Bannayan-Riley-Ruvalcaba (BRR) Syndrome. PMID- 19813503 TI - AMA and Wisconsin Medical Society: together we are stronger. PMID- 19813504 TI - Integration of spine research and clinical activities drives innovation, outcomes. PMID- 19813505 TI - A project to reduce potentially inappropriate medications and drug interactions. PMID- 19813506 TI - Sentinel events, disruptive behavior, and medical staff codes of conduct. AB - Regardless of physician concerns, hospitals and other organizations accredited by the Joint Commission are required to comply with the new leadership standard and define acceptable, disruptive, and inappropriate behaviors in a code of conduct. The new standard also requires them to implement a process for managing disruptive and inappropriate behaviors. Rules and standards can be very effective ways to promote safety and quality, but at the same time may be subject to abuse if not properly monitored. As a result, physicians should continue to monitor how hospitals have implemented the new leadership standard and raise concerns about any vague definitions, encroachment on physician rights, and misuse of the code of conduct. PMID- 19813507 TI - Hawaii DENTAC conducts humanitarian dental missions to remote locations. PMID- 19813508 TI - Networking within your professional association. PMID- 19813509 TI - Treating the special needs patient with a sensory disability: hearing impairment. PMID- 19813511 TI - Patient perceptions: the dental staff's influence on patient satisfaction. PMID- 19813510 TI - Managing cultural differences in the dental office. PMID- 19813512 TI - H.R. 3200--an attempt at health reform. PMID- 19813513 TI - Stark Law changes are on the horizon. PMID- 19813514 TI - Field evaluation and management of non-battle related knee and ankle injuries by the advanced tactical practitioner (ATP) in the austere environment--Part Two. PMID- 19813515 TI - Canine tactical field care. Part two--Massive hemorrhage control and physiologic stabilization of the volume depleted, shock-affected, or heatstroke-affected canine. AB - Military and law enforcement agencies have seen a dramatic increase in the utilization of military working dogs (MWDs) and working canine officers, respectively both at home and in foreign deployments. Due to the fact that professional veterinary care is often distant from internal disaster or foreign deployment sites, the military medic, police tactical medic, or other first response medical care providers may be charged with providing emergency or even basic, non-emergency veterinary care to working canines. The medical principles involved in treating canines are essentially the same as those for treating humans; however, the human healthcare provider needs basic information on canine anatomy and physiology, and common emergency conditions, in order to provide good basic veterinary care until a higher level of veterinary care can be obtained. This article represents the second in a series designed to provide condensed, basic veterinary information on the medical care of working canines, including police canines, federal agency employed working canines, and search-and-rescue dogs, in addition to the MWD, to those who are normally charged with tactical or first responder medical care of human patients. This article focuses on diagnosing and treating some of the more common high-mortality conditions affecting canines in the field including massive hemorrhage, volume-depletion, shock, and heatstroke. PMID- 19813516 TI - A case of reactive arthritis in a Ranger Indoctrination Program (RIP) student. AB - Musculoskeletal complaints comprise the majority of cases encountered by military physicians when evaluating young active duty Soldier-athletes. This is a case of reactive arthritis in a 19-year-old active duty Soldier-athlete whose failure to improve with conservative therapy initiated further investigation. When evaluating what appear to be routine overuse injuries, it is important to actively include other potential causes of musculoskeletal complaints in the differential diagnosis. Further investigation of disease in patients whose symptoms and complaints do not improve with routine conservative care is paramount. Reactive arthritis, though self-limiting in two-thirds of those affected, can become a chronic disabling disease affecting as many as 40 out of 100 patients. Current theories suggest the persistent presence of non-culturable bacteria and bacterial antigens residing in the joint synovia as the etiology of the disease state. There is no curative therapy for reactive arthritis and management is focused on the treatment of symptoms with non-steroidal anti inflammatory drugs (NSAIDs), immunomodulator therapy, and antibiotics if an infectious source is suspected. PMID- 19813517 TI - Functional training program bridges rehabilitation and return to duty. AB - Traditional clinic-based rehabilitation programs often fall short of returning Soldiers to peak condition prior to releasing them for duty. With the higher physical demands placed on the Special Operations Soldier, a bridge program offers rehabilitation professionals a way to maximize recovery, enhance performance, and hopefully prevent injuries (or re-injury). A six week functional training program is outlined and data collection from over two years is presented. Statistically and operationally significant differences were noted in nearly every category tested. Functional Movement Screen scores improved an average of 2.5 points. T-test improvement was 0.5 seconds. Single leg hop time improved 10%. Hop for distance improved approximately 10%. Body fat improvement was statistically significant. Kip-ups improved 32%. Vertical jump height improvement was statistically significant. All subjective fitness category self evaluations demonstrated statistically significant improvements, except for pain. Data suggests that a program like this may be beneficial to patients and non patients seeking a safe, effective alternative training regimen. PMID- 19813518 TI - The impedance threshold device (ITD-7)--a new device for combat casualty care to augment circulation and blood pressure in hypotensive spontaneously breathing warfighters. AB - Inspiration through -7cm H2O resistance results in an increase in venous blood flow back to the heart and a subsequent increase in cardiac output and blood pressure in hypotensive animals and patients. Breathing through the impedance threshold device with 7cm H2O resistance (ITD-7) also reduces intracranial pressure with each inspiration, thereby providing greater blood flow to the brain. A new device called an ITD-7 was developed to exploit these physiological mechanisms to buy time in hypotensive War Fighters when other therapies are not readily available. Animal and clinical data with the ITD-7 demonstrate the potential value and limitations of this new non-invasive approach to enhancing circulation. PMID- 19813519 TI - The United States Army Special Forces--Walter Reed Army Institute of Research Field Epidemiologic Survey Team (Airborne). AB - The U.S. Army Special Forces--Walter Reed Army Institute of Research Field Epidemiological Survey Team (Airborne) was formed in late 1965 and later deployed to Vietnam in 1966. Funded by Walter Reed Army Institute of Research and staffed by highly trained Special Forces qualified medical personnel from Fort Bragg, North Carolina, the team was attached to the 5th Special Forces Group (Airborne) while in Vietnam. During its short existence, the team conducted extensive and important field studies on diseases of military medical importance, often under combat conditions. PMID- 19813520 TI - SORT(ing) out the casualties: the Special Operations Resuscitation Team in Afghanistan. PMID- 19813521 TI - Baseline dissociation and prospective success in Special Forces Assessment and Selection. AB - INTRODUCTION: Although dissociation at the time of trauma (peritraumatic dissociation) has been shown to predict the development of posttraumatic stress disorder (PTSD), it is not yet known whether the tendency to dissociate under nonstressful circumstances (i.e., at baseline) can also serve as a predictor of vulnerability to stress in healthy individuals. METHOD: Baseline symptoms of dissociation (CADSS) were assessed in 774 active duty male Soldiers enrolled in Special Forces Assessment and Selection (SFAS). RESULTS: Soldiers who endorsed experiencing any symptoms of dissociation at baseline were significantly less likely to be successful in SFAS. The greater the number of symptoms of dissociation endorsed at baseline, the greater the likelihood of failure. DISCUSSION: These data explain our earlier findings of fewer symptoms of dissociation in elite troops and may have relevance for the selection and hiring of personnel for nonmilitary, at-risk professions. Better screening may lead to improved primary intervention strategies, better job placement, and lowered risk of PTSD. PMID- 19813522 TI - Prehospital advances in the management of severe penetrating trauma. AB - BACKGROUND: Historic advances in combat prehospital care have been made in the last decade. Unlike other areas of critical care, most of these innovations are not the result of significant improvements in technology, but by conceptual changes in how care is delivered in a tactical setting. The new concept of Tactical Combat Casualty Care has revolutionized the management of combat casualties in the prehospital tactical setting. DISCUSSION: The Tactical Combat Casualty Care concept recognizes the unique epidemiologic and tactical considerations of combat care and that simply extrapolating civilian care concepts to the battlefield are insufficient. SUMMARY: This article examines the most recent and salient advances that have occurred in battlefield prehospital care driven by our ongoing combat experience in the Iraq and Afghanistan and the evolution around the Tactical Combat Casualty Care concept. PMID- 19813523 TI - Results of Vietnamese acupuncture seen at the Second Surgical Hospital. AB - One of the rewards in the medical service in a foreign land is the exposure to exotic diseases and methods of therapy which are virtually unknown to many physicians in the western world. With the treatment of Vietnamese patients by medical personnel from the United States and other countries with a similar system of medical education, new experiences and challenges are presented daily. The use of acupuncture as a form of therapy is undoubtedly as unfamiliar to many physicians as it was to us. In addition to our treatment of American casualties at the Second Surgical Hospital in the Republic of Vietnam, we had the opportunity of treating some interesting problems in Vietnamese patients who had previously been treated with acupuncture. The majority of our own staff questioned, "What is acupuncture?" PMID- 19813524 TI - Overview of combat trauma in military working dogs in Iraq and Afghanistan. PMID- 19813525 TI - Alert--watch for "look alike" Combat Application Tourniquet (C.A.T.). PMID- 19813526 TI - Skill identifier for Army Medical Department officers in Special Operations. PMID- 19813527 TI - Charmak disease in Afghanistan. PMID- 19813528 TI - Picture this...sarcoidosis. PMID- 19813529 TI - [Demented patient with strangulation marks. Was it a crime?]. PMID- 19813530 TI - [Food poisonings are increasing. Dangerous grilled meat (interview by Dr. Judith Neumaier)]. PMID- 19813531 TI - [Germans have an unhealthy life style. How family physicians can make improvements]. PMID- 19813532 TI - [Future prognosis. Germany will be this sick in 40 years]. PMID- 19813533 TI - [Chronic coronary heart disease. Family practice therapy is restored]. PMID- 19813534 TI - [National practice guideline for diabetic foot syndrome. Treatment methods to forestall amputation]. PMID- 19813535 TI - ["Lung problems" since childhood. Surprise in the thoracic x-ray]. PMID- 19813536 TI - ["...Men are so vulnerable ..." does our medicine do justice to men?]. PMID- 19813537 TI - [Depression in men: stress, risk factors and symptoms]. PMID- 19813538 TI - [Aspects of psychosomatic medicine and current treatment options in male sexual dysfunction]. PMID- 19813539 TI - [Emergency checklist: supraventricular tachycardia]. PMID- 19813540 TI - [Diagnosis, treatment and rehabilitation of peripheral facial nerve paralysis]. PMID- 19813541 TI - [Fibromyalgia syndrome: diagnostic criteria and treatment]. PMID- 19813542 TI - [New thrombocyte inhibitor. Fewer ischemic complications in acute coronary syndrome]. PMID- 19813543 TI - [Anticoagulation in atrial fibrillation. Stroke can be prevented this way]. PMID- 19813544 TI - Unlock the doors. PMID- 19813545 TI - Marking the passage of time. PMID- 19813546 TI - Design for Health and Human Services showcase. PMID- 19813547 TI - Selecting durable and safe furniture. PMID- 19813548 TI - A fresh start for homeless services in Miami. PMID- 19813549 TI - A new plan for patients in crisis. PMID- 19813550 TI - Building on history at Eastern State Hospital. PMID- 19813551 TI - Following the evidence toward better design. PMID- 19813553 TI - Patient safety in obstetrics and beyond. PMID- 19813552 TI - The 42 CFR Part 2 and NHIN conundrum. PMID- 19813554 TI - Interview with a quality leader: Paul Gluck, immediate past chair, National Patient Safety Foundation. Interview by Pamela K Scarrow. AB - Dr. Paul Gluck, MD, FACOG, has held many leadership positions. He served as the president/chair of the William A. Little OB/GYN Society, the Miami OB/GYN Society, the Florida OB/GYN Society, the Baptist Health Foundation, the Health Council of south Florida, the Florida Section of the American College of OB/ GYN (ACOG), National Patient Safety Foundation, as well as the Dade County Medical Association. He is currently ACOG assistant secretary and serves on their Executive Committee. Dr. Gluck has an interest in access to healthcare. For his work in establishing a prenatal clinic in an area of critical need he received ACOG president's Service Award and Humanitarian of the Year Award from the South Florida Perinatal Network. He led the Florida initiative to promote depression screening and treatment in women recognized by the Wyeth National Section Award. He co-chaired the Governance Committee of the Mayor's Task Force charged with solving the problem of providing care for the over 450,000 uninsured residents of Miami-Dade County. PMID- 19813555 TI - Advancing patient safety through process improvements. AB - The department of Women's and Children's Services at the Hospital of Saint Raphael (HSR) in New Haven, CT, has initiated several different and successful approaches to reducing patient risk within the department. The department purchased a computerized fetal monitoring and documentation program that has improved the ability to provide high-level antepartal care for mothers and fetuses with automatic patient data management and continuous fetal heart rate surveillance. A Risk Reduction Grant offered through the hospital malpractice insurance program provided the financial assistance for all medical providers to become certified in electronic fetal monitoring. The certification is now a required educational standard for nurses, certified nurse midwives, and for physicians who work in the labor and delivery unit. Infant and pediatric security is incorporated into policy and practice measures that include hospital-wide drills for the prevention of infant abduction. The Obstetrics and Gynecology (OB/GYN) Quality Improvement Committee supports systematic reviews of identified clinical risks and works to find viable solutions to these problems. The hospital has supported specialized obstetrical care through the Maternal Fetal Medicine Unit (MFMU), Newborn Intensive Care Unit (NICU), the Inpatient Pediatric Unit, and the labor and delivery unit. In addition, HSR has initiated an enhanced medical informed consent that is available online for providers and a patient education tool that includes a computer room at the hospital for patient use. PMID- 19813556 TI - Teamwork: crew resource management in a community hospital. AB - Effective communication and teamwork among the healthcare team has a profound impact on patient care; in the Delivery Room it means the best possible outcome for moms and babies. With that goal in mind, our obstetrics team, partnered with Harvard Risk Management Strategies Foundation Team Performance Plus (TPP), implemented an initiative in May 2006. The primary goal of the initiative was to improve patient care through improved communication between disciplines, situation monitoring throughout the continuum of care, mutual support and respect among care givers, and effective team leadership. Through education and implementation of specific communication tools and behaviors, we realized better patient outcomes as well as improved patient and staff satisfaction. PMID- 19813557 TI - Collaborative communication: integrating SBAR to improve quality/patient safety outcomes. AB - Creating work environments that sustain open and supportive communication positively influence teamwork, staff satisfaction, and improved patient quality and safety. The Situation, Background, Assessment, and Recommendation (SBAR) collaborative communication evidence-based practice (EBP) study described in this article introduced collaborative communication integrating SBAR communication process in a pediatrics/perinatal services department of a 271-bed community hospital in northern Arizona. EBP processes framed the study. Evaluation methods for intervention effectiveness and study outcomes integrated both quantitative and qualitative strategies. Staff transferred evidence, knowledge, and skills into practice to achieve enhanced communication, collaboration, satisfaction, and patient safety outcomes meeting the study goal. PMID- 19813558 TI - Back to basics: creating a simulation program for patient safety. AB - As the malpractice crisis in Obstetrics continues to spiral, safety initiatives within Obstetrics have grown to include simulation as a primary risk reduction strategy. Starting a simulation program requires careful planning and the commitment of all nursing and physician providers to promote a culture of patient safety. This article discusses eight recommended steps towards implementing a simulation program and illustrates through a case scenario how the process of building such a program will be integral to both its success and the fulfillment of your department's goals of providing excellent patient care. PMID- 19813559 TI - Integrating team training strategies into obstetrical emergency simulation training. AB - Successful management of obstetrical emergencies such as shoulder dystocia requires the coordinated efforts of a multidisciplinary team of professionals. Simulation education provides an opportunity to learn and master simple as well as complex technical skills needed in emergent situations. Team training has been shown to improve the quality of communication among team members and consequently has an enormous impact on human performance. In the healthcare environment, especially obstetrics where the stakes are high, integrating team training into simulation education can advance efforts to create and sustain a culture of safety. With over 7,100 deliveries annually, our 1,100-bed, two-hospital regional healthcare system embarked on this journey to advance the culture of safety. PMID- 19813560 TI - The unintended consequences of being friendly: a case study. AB - Baby-friendly certification recognizes hospitals that promote early physical bonding between mother and infant, immediately after birth. Most births can accomplish physical bonding without increased risk to mother or infant. When mother or infant have complications and each have intravenous (i.v.) lines and are receiving medications, the physical bonding post-birth may also inadvertently put the patients at risk. A baby-friendly community hospital in New England found that early bonding put an infant at higher risk for medication error when the two i.v. lines were not properly identified and the infant received a medication intended for the mother. The growing body of literature on i.v. medication safety does not address this particular type of error, and this was an error that technology would not have prevented. The "5 rights" of medication safety are not as effective as physical separation of the two individuals during medication administration. A brief separation does not diminish bonding, and the practice has prevented subsequent errors. PMID- 19813561 TI - Virtual patient safety rounds: one hospital system's approach to sharing knowledge. AB - Understanding how and why errors in healthcare happen is essential to improving patient safety. Yet exposure to this learning process is usually limited to those events occurring in one's own institution. Virtual Safety Rounds expands this learning opportunity to multiple hospitals. Twice each month physicians; nurses; and quality, risk, and patient safety staff participate in a discussion about a recent safety event within the healthcare system. Within this safe collegial environment experiences, plans of correction and lessons learned are shared. Hospitals are learning from each other without having to experience the patient safety issue directly. PMID- 19813562 TI - CCHIT & the DMV. Certification is moving in the right direction, but perils lay around every corner. PMID- 19813563 TI - Can high tech happen on a low budget? PMID- 19813564 TI - "It's not my problem" (yet). With readmissions under the microscope, CIOs know chronic disease management is moving front and center. AB - Chronic disease management is a system initiative, not just for primary care or ambulatory. The business case is currently poor for chronic disease management, but may soon change due to reform. Chronic disease management can help prevent costly readmissions. Care management plans are often a component of an EMR. PMID- 19813565 TI - All the help you can get. KLAS' "2009 Mid-Term Performance Review: Software and Professional Services" offers a clear way to see if your main vendor is moving in the right direction. PMID- 19813566 TI - Watching, waiting & studying. While HITECH has providers studying their KLAS reports, a lack of clarity on meaningful use means few are pulling the trigger. PMID- 19813567 TI - Getting together. As changes in policy and meaningful use are made, decisions on financial systems increasingly depend on integration with clinical. PMID- 19813568 TI - One-on-One with KLAS President Adam Gale. Gale discusses a HITECH buying frenzy that wasn't, & why Epic's page of the Mid-Term Performance Review makes competitors green with envy. Interviewed by Anthony Guerra. PMID- 19813569 TI - Has HITECH hijacked Stark? Now that physicians can get reimbursed for EMRs directly from the government, CIOs are reexamining their Stark strategy. AB - HITECH is offering physicians $44,000 for an EMR. Most hospitals have been slow to take advantage of the Stark relaxations. Lowering the Stark subsidy to physicians can weed out those who aren't really motivated to implement an EMR. Most feel the number of physician practices declining hospital Stark funds will be small. Any Stark budget funds not used should be re-invested in linking the hospital and the practices. PMID- 19813570 TI - Right on schedule. While there are no easy wins in IT, some say automated staff scheduling systems come close. AB - Automated systems can house schedules from different units and facilities in a centralized location, giving managers the visibility they need to more effectively allocate resources. Smart CIOs are leveraging the business intelligence tools in scheduling systems to better anticipate staffing needs based on trends. With Web-based staffing, nurses can control their schedules by viewing openings in other facilities and offering to fill shifts. Many hospitals tie in incentives to encourage "floating" to other hospitals within a system. While some hospitals use scheduling as a stand-alone system, others have found value in linking it to other systems like time and attendance, HR and payroll. PMID- 19813571 TI - Cutting the cord. CIOs are leveraging wireless technologies to help nurses deliver patient care safely and efficiently. AB - The proliferation of wireless technologies in the hospital setting has significantly impacted the way nurses deliver care. The primary goals ClOs are looking to achieve in implementing wireless technologies are to improve communication, workflow efficiency and patient safety. Organizations that have successfully implemented wireless technologies have had nursing leaders involved in every key step of the process. PMID- 19813572 TI - The heart of PACS. While most CIOs feel secure taking on the challenge of traditional radiology-focused PACS, cardiology PACS is another story. AB - In terms of both workflow and the type of specialty care involved, cardiology PACS is inherently more complex and challenging than radiology PACS. Very few organizations have gotten beyond some initial work in automating the collection of data from one or a few different clinical processes. Even at this early phase of development, it's clear that clinician workflow and efficiency can be dramatically improved by cardiology PACS implementation. Enterprise-wide image and data storage, retrieval, and sharing capability across cardiology, radiology, and other specialties, is where hospital organizations are headed, though the path is a long and difficult one. PMID- 19813573 TI - Is the bar still too high? ONCHIT appeased some by tweaking CPOE requirements, but many still say the timeline for patient access to records is too aggressive. PMID- 19813574 TI - For all the right reasons. Approaching CPOE from a patient safety and care quality perspective is the first critical step toward success. AB - True CPOE success is about facilitating improved patient safety, care quality, and efficiency in a multidisciplinar environment, and on an ongoing basis. CPOE implementation forces clinician leaders to examine and rework long-ingrained care delivery processes, especially as they build or adapt order sets. The likelihood that CPOE will be a requirement of meaningful use could compel a rapid acceleration in implementation. PMID- 19813575 TI - It's all about workflow. A misplaced focus on automating transactions will never bring about the healthcare system everyone seeks. PMID- 19813576 TI - Smoothing the path. Leading with portals can lay the groundwork for CPOE, making rollouts less risky. PMID- 19813577 TI - Voice of reason. Honest communication can help keep staff grounded, focused and inspired through difficult times. PMID- 19813578 TI - Treat-grandfather of CPOE. While high costs put cell phones out of reach, one vendor was selling an order entry system ahead of its time. PMID- 19813579 TI - A model asthma program. PMID- 19813580 TI - Coding & billing. Capturing revenue, avoiding coding errors take on higher priority. PMID- 19813581 TI - Patient care. Spread of infectious diseases may run high with patient sharing. PMID- 19813582 TI - Technology. Ambulances gain access to vital medical records. PMID- 19813583 TI - Planning for a pandemic. Interview by Haydn Bush. PMID- 19813584 TI - Hospital staff goes into overdrive during anthrax scare. PMID- 19813585 TI - Roxie's excellent adventure. PMID- 19813586 TI - Physician alignment. PMID- 19813587 TI - Building the medical home. AB - All the talk about bundled payments or otherwise paying for care across the continuum has put the spotlight on the medical home model. Challenges abound. PMID- 19813588 TI - Connecting all your docs. PMID- 19813589 TI - 2009 quest for quality award winners. PMID- 19813590 TI - AHA environmental scan 2010. PMID- 19813591 TI - Peer perspective deepens. PMID- 19813592 TI - [A hybrid reconstruction method in electrical impedance tomography based on GMRES and Tikhonov regularization]. AB - Electrical impedance tomography (EIT) is a technique for reconstructing the conductivity distribution of measured field owing to its characteristics of being non-homogeneous, of injecting current at the boundary of the measured subject,of measuring the.corresponding changes in voltage, and of reconstructing the image of the subject consequently. However, the limited measurement data of EIT, and the serious nonlinearity of the field result in ill-posed problem, and the resolution of reconstructed image is poor. To solve the problem, a new hybrid algorithm is herein proposed. The method combines the characteristics of Krylov subspace and Tikhonov regularization, which can improve the real time performance, the quality and robustness of reconstructed image. PMID- 19813593 TI - [Computer-aided surgery planning for implantation of artificial ear]. AB - In conventional ear implantation surgery, clinical physicians usually make a surgery planning based on their observation on series of 2D X-ray images or CT images. Such a planning method requires the physicians to have a high level of clinical experience. Besides, the whole operation is unintuitive, and might have certain risk. Considering these facts, we have developed a computer-aided system for the surgery planning of the implantation of artificial ear based on CT imaging and 3D reconstruction techniques. The system effectively overcomes the main drawbacks in conventional surgery planning techniques, and it makes the surgery planning procedure more precise, safe, and intuitive. PMID- 19813594 TI - [An improved medical image fusion algorithm and quality evaluation]. AB - Medical image fusion is of very important value for application in medical image analysis and diagnosis. In this paper, the conventional method of wavelet fusion is improved,so a new algorithm of medical image fusion is presented and the high frequency and low frequency coefficients are studied respectively. When high frequency coefficients are chosen, the regional edge intensities of each sub image are calculated to realize adaptive fusion. The choice of low frequency coefficient is based on the edges of images, so that the fused image preserves all useful information and appears more distinctly. We apply the conventional and the improved fusion algorithms based on wavelet transform to fuse two images of human body and also evaluate the fusion results through a quality evaluation method. Experimental results show that this algorithm can effectively retain the details of information on original images and enhance their edge and texture features. This new algorithm is better than the conventional fusion algorithm based on wavelet transform. PMID- 19813595 TI - [A study on the four modes for transmitting heart sound signal]. AB - As an important human body sound signal, heart sound is of great value in the researches on diagnostics of heart diseases. This study sought to explore the methods of transmitting heart sound through the telephone correspondence system for simultaneous telemetering cardiac contractility and heart rate. Heart sounds were transmitted from a phone to another phone with 4 modes, the wirelessly transmitted distance between the two phones being 5 m, 10 km, and 1000 km, respectively. The results of experiments show that telemetering cardiac contractility and heart rate can be realized by the telephone correspondence system. Such methods have the advantages of being noninvasive, inexpensive, rapid and convenient; moreover, they can be used repeatedly and be available for in home use. PMID- 19813596 TI - [A method of dental arch auto-detection for dental plaster models]. AB - The shape of dental arch for orthodontic diagnosis and treatment is of great significance. This paper presents an automated method for detecting the dental arch form. Firstly, 3D teeth data model is retrieved by the 3D-optical measuring system. Secondly, the occlusal plane is computed by interactively picking up four feature points. Thirdly, the feature point set is filtered by the rule and two step curve fitting method is used to obtain the dental arch form. Finally, some examples are tested in this work and the results demonstrate that the proposed algorithm is effective and feasible. PMID- 19813597 TI - [A novel image processing and analysis system for medical images based on IDL language]. AB - Medical image processing and analysis system, which is of great value in medical research and clinical diagnosis, has been a focal field in recent years. Interactive data language (IDL) has a vast library of built-in math, statistics, image analysis and information processing routines, therefore, it has become an ideal software for interactive analysis and visualization of two-dimensional and three-dimensional scientific datasets. The methodology is proposed to design a novel image processing and analysis system for medical images based on IDL. There are five functional modules in this system: Image Preprocessing, Image Segmentation, Image Reconstruction, Image Measurement and Image Management. Experimental results demonstrate that this system is effective and efficient, and it has the advantages of extensive applicability, friendly interaction, convenient extension and favorable transplantation. PMID- 19813598 TI - [The development of a wearable pulse oximeter sensor and study of the calibration method]. AB - The paper first analyses the principles of measurement of the two-wave oximeter and their limitations in technology. We propose to filter off motion interference from pulse oximeter signal using an algorithm based on the Beer-Lambert law that requires a three-wave probe (660 nm, 850 nm, and 940 nm). Based on the new algorithm, this paper describes the design principle of the circuitry and the software flowchart. Also, we study the calibration method of the pulse oximeter sensor and discuss the results in this paper. PMID- 19813599 TI - [Research of liver tumor extraction based on transparency]. AB - It is very difficult to use the conventional methods that are based on area and on border for extracting a region of intered such as liver tumor region with vague and irregular boundaries. This paper introduces Poisson matting that uses transparency (alpha value) as self-property of image and seeks its best result value to achieve the aim of extracting object. Meanwhile, this method is applied to the liver tumor CT picture experimental results. It makes improvement in two steps: computing image source a value; calculating foreground image F and background image B. Consequently, successful result is obtained. PMID- 19813600 TI - [Entropy analysis of the EEG alpha activity in depression patients]. AB - The alpha rhythm in electroencephalograph has been associated with the mental state of the brain. The aim of the present study was to analyze the complexity of the alpha activity in depression patients by Renyi entropy (RE) under two conditions, i.e., resting with eyes closed followed by a simple mental arithmetic. The EEGs were recorded in 20 depression patients and 20 age-matched control subjects. Results showed that the alpha activity of depression patients is more complex during resting and is more regular during the mental arithmetic than that of the controls. RE analysis can well characterize the alpha activity in depression patients and could be a valuable tool to study cognitive process and also a potential effective approach for the diagnosis of depression. PMID- 19813601 TI - [Comparison of digital filter and wavelet transform for extracting electroencephalogram rhythm]. AB - It is very important to extract electroencephalogram (EEG) rhythm in clinical diagnoses. Digital filter and wavelet transform are used to extract the rhythm from a piece of EEG at the sampling rate of 2 kHz. The Daubechies order 4 wavelet (db4) was used to decompose the EEG at 8 levels. According to the filter characteristic of wavelet decomposition, the reconstructions of aS, d8, d7, d6 and d5 component are nearly corresponding to the rhythms of delta, theta, alpha, gamma separately. The 6 order ellipse infinite impulse response (IIR) filter is also used to decompose the EEG. As the quality factor of wavelet decomposition filter is constant, the wavelet transform obtains better extracted rhythm than the digital filter. Furthermore, the wavelet transform method can be used to extract the low frequency rhythm from wide frequency band. PMID- 19813602 TI - [Detection of QRS complexes using wavelet transformation and golden section search algorithm]. AB - The extraction and identification of ECG (electrocardiogram) signal characteristic parameters are the basis of ECG analysis and diagnosis. The fast and precise detection of QRS complexes is very important in ECG signal analysis; for it is a pre-requisite for the correlative parameters calculation as well as for correct diagnosis. In our work, firstly, the modulus maximum of wavelet transform is applied to the QRS complexes detection from ECG signal. Once there are mis-detections or missed detections happening, we utilize the Golden Section Search algorithm to adjust the threshold of maxima determination. The correct detection rate of the QRS complexes is up to 99.6% based on MIT-BIH ECG data. PMID- 19813603 TI - [The pulmonary arterial oxygen saturation monitoring via trachea in hypoxia dogs]. AB - This study was aimed to assess the agreement between pulse pulmonary arterial oxygen saturation (StO2) by oximetry in trachea and the mixed venous oxygen saturation (SvO2) by fiberoptic pulmonary arterial catheter, and to evaluate the accuracy of StO2 monitoring during hypoxia. 10 mongred dog's were used. After anesthesia was induced and thorax was opened, we placed a fiberoptic pulmonary artery catheter directly through the dog's right ventricular outlet. Then, we placed and adjusted the trachea catheter, attached the oximetry probe to trachea carina till the high quality StO2 PPG signal was obtained, and till the readings were stable at about +/-2% from fiberoptic catheter. The pair readings of StO2 and SvO2 were recorded at every minute interval in 10 minutes when circulation was stable. Decreasing the inhaled oxygen concentration till the tongue's SpO2 decreased to 60%; recording the changes of StO2 and SvO2 in every 5% drop of tongue's SpO2 when the tongue's SpO2 decreased from 100% to 50%. The results showed that there was a good agreement between the two methods for pulmonary arterial oxygen saturation measurement. However, the difference between the two methods was great and unacceptable during the presence of hypoxia. PMID- 19813604 TI - [Effects of static magnetic fields on aerobes: Escherichia coli, Staphylococcus aureus and Bacillus subtilis]. AB - Nutrient agar medium was exposed in 0.085-0.092 T static magnetic fields for 12 h. Then we densities the optical densities at lamda = 600 (OD600) of Escherichia coli, Staphylococcus aureus and Bacillus subtilis in different culturing stage. The results were compared with those of control group in the normal geomagnetic field. The OD600 values of experimental groups of these three kinds of aerobes were significantly higher than those of control groups from 3h to 9h. However, after 11 h, there was no remarkable difference regarding the OD600 values between the two groups. The dissolved oxygen content of nutrient agar medium was determined by microtitration. The dissolved oxygen of nutrient agar medium under static magnetic for 12h increased 15% in average and there was significant difference when compared with the control. The results showed that the ferro magnetic fields increased the dissolved oxygen content of nutrient agar medium significantly. These findings suggest that the effects of static magnetic fields on Escherichia coli, Staphylococcus aureus and Bacillus subtilis are related to the dissolved oxygen. PMID- 19813605 TI - [Improvement on high frame rate ultrasonic imaging system based on linear frequency-modulated signal]. AB - The high frame rate (HFR) ultrasonic imaging system based on linear frequency modulated (LFM) signal constructs images at a high frame rate; the signal-to noise ratio (SNR) of this system can also be improved. Unfortunately, such pulse compression methods that increase the SNR usually cause range sidelobe artifacts. In an imaging situation, the effects of the sidelobes extending on either side of the compressed pulse will be self-noise along the axial direction and masking of weaker echoes. The improvement on high frame rate ultrasonic imaging system based on LFM signal is considered in this paper. In this proposed scheme, a predistorted LFM signal is used as excited signal and a mismatched filter is applied on receiving end. The results show that the proposed HFR ultrasonic imaging system can achieve higher SNR and the axial resolution is also improved. PMID- 19813606 TI - [A modified fast independent component analysis and its application to ERP extraction]. AB - Feature extraction of event-related potentials (ERP) plays an important part in both basic and clinical researches for cerebral neurophysiology. ICA is a method for separating blind signals based on signal statistic characteristics. In this paper, the fundamental principle, the discrimination condition and the practical algorithm of Independent Component Analysis are discussed. Then, a fast Independent Component Analysis algorithm (Fast ICA) is introduced. But like Fast ICA, its convergence is dependent on initial weight. We bring in a revision factor into the algorithm; thus the new algorithm could implement convergence on a largescale. In this paper, the revision factor is calculated by gradient. By modifying kernel iterate course, several iterations of Fast ICA are merged into one iteration of Modified Fast ICA, so the convergence of ICA will be accelerated. Finally, Modified ICA is applied to ERP extraction. The simulation shows that the convergence speed can be increased by using the improved algorithm. PMID- 19813607 TI - [Bone remodelling model including mechanism of damage and repair]. AB - From the visual angle of mechanics, bone remodelling plays an important role in adapting to load environment and repairing microcracks. In order to describe the process of bone remodelling explicitly, we introduced a bone remodelling model representing the biological features of damage/repair in bone. Based on Basic Multicellular Units (BMUs), the model embodies not only the mechano-transduction of osteocytes' response to mechanical stimuli, but also the biological process of the randomly ocurring microcracks that are resorbed by osteoclasts and then refilled by osteoblasts. A 2-D trabecular bone finite element model was used as an initial configuration in the research. The developments in bone microstructure and material properties have been monitored in the course of 10 simulated years. The conclusions are: (1) Initially, trabecular bone changes prominently, but it tends to be stable later on; (2) The material properties of trabecular bone in compression are much better than in tension. PMID- 19813608 TI - [Preliminary observation of the blocking effect produced by injecting polyacrylamide hydrogel on low hydraulic resistance channel along meridian]. AB - To study the role of meridians in the formation and development of diseases, a pathological model of obstructed channel was established by injecting polyacrylamide hydrogel. The effects of blocking low hydraulic resistance channel (LHRC), produced by injecting polyacrylamide hydrogel and by injecting normal saline, were compared by examining the change in transmission of interstitial fluid pressure wave. The results showed that there was significant decrease (P < 0.01) in interstitial fluid pressure wave after more than 0.5 ml polyacrylamide hydrogel was injected into the channel, whereas no significant changes were found after normal saline was injected or when the hydrogel was outside the channel. The above findings demonstrate that the low hydraulic resistance channel can be blocked by injecting certain amount of polyacrylamide hydrogel and a pathological model of obstructed channel has been established preliminarily. PMID- 19813609 TI - [Effects of alcohol on bone metabolism and biomechanical property of mice]. AB - This study was aimed to detect the effects of alcohol on bone metabolism and biomechanical property of growing mice. Thirty KM mice were randomly divided into 3 groups, namely basal control group (mice were killed at the beginning), normal control group (with distilled water given by gastrogavage), and 50% (V/V) alcohol group (with alcohol given by gastrogavage at the dose of 4 g x kg(-1) x d(-1) for 60 days). All mice were killed and their proximal tibia and tibial diaphysis were processed by undecalcified sections and measured by bone histomorphometry. The biomechanical properties of lumbar vertebra and femur were tested. Compared with normal control, the index of trabecular bone area (% Tb. Ar) of proximal tibial metaphysis (PTM) and the static parameter of cortical bone( Ct. Ar) both decreased obviously (P < 0.05) in alcohol group. Bone formation rate (BFR/TV) of trabecular bone and cortical bone dropped also (P < 0.05). The maximal resistibility of lumbar vertebra and structural mechanical strength of proximal femoral neck both declined significantly (P < 0.01) in alcohol group. Low dose of alcohol inhibited the bone formation rate of growing mice , thus leading to a disorder of bone metabolism and a decrease in biomechanical quality. PMID- 19813610 TI - [The corrosion of pure iron in five different mediums]. AB - The sectional test was adopted in this study to investigate the corrosion of pure iron in 0.15 mol/L NaCl solution, Ringer solution, PBS(-) solution, SBF solution and M199 cell culture medium at three different times. The result shows that different mediums have different corrosion effects on pure iron. The arrangement according to the medium's corrosion ability from the strongest to weakest is 0.15 mol/L NaCl solution (Ringer solution), PBS(-) solution, SBF solution and M199 cell culture medium. The results of scanning electron microscopy and energy dispersive X-ray spectrum analyses show that the addition of HPO4(2-), H2POC4-, Ca2+, Mg2+, SO4(2-) and the organic component can inhibit the corrosion to some degree. PMID- 19813611 TI - [Effect of SspA on the formation of bacterial biofilm covering the surfaces of cardiovascular biomaterial Dacron]. AB - This study sought to assess the effect of SspA on the formation of Staphylococcus aureus biofilm extending over the surfaces of Cardiovascular Biomaterial Dacron. SspA was extracted from the surface of staphylococcus aureus biofilm, purified, and then used to influence the adhesion of Staphylococcus aureus and the formation of Staphylococcus aureus biofilm on Dacron biomaterial surfaces. The formation of the Staphylococcus aureus biofilm on cardiovascular biomaterial Dacron surfaces under gradient SspA concentrations was evaluated by confocal laser microscopy. The result revealed that SspA inhibited the formation of Staphylococcus aureus biofilms on cardiovascular biomaterials surfaces effectively, and it was dose dependent. This study indicates that SspA is effective for preventing biomaterial centered infection and this method is conducive to clinical applications. PMID- 19813612 TI - [Effects of silicon carbide on the cure depth, hardness and compressive strength of composite resin]. AB - The hardness, compressive strength and cure depth are important indices of the composite resin. This investigation was made with regard to the effects of silicon carbide on the cure depth, hardness and compressive strength of the light curing composite resin. Different amounts of silicon carbide were added to the light-curing composite resin, which accounted for 0 wt%, 1 wt%, 0.6 wt%, 0.3 wt%, 0.1 wt%, 0.05 wt% and 0.005 wt% of the composite resin, respectively. The hardness, compressive strength and cure depth of the six afore-mentioned groups of composite resin were measured by the vernier caliper, the vickers hardness tester and the tensile strength of machine, respectively. The results showed that silicon carbide improved the hardness and compressive strength of the light curing composite resin,when the concentration was 0.05 wt%. And the cure depth was close to that of control. PMID- 19813613 TI - [Preparation of porous Ti metal composite scaffold with bioactivity]. AB - In this work, the titanium powder was used for preparing highly interconnected porous scaffolds by impregnating polymer method. Subsequently, the electrochemical method and the biomimetic mineralization method were adopted to deposit calcium phosphate coatings on the sintered scaffold which was supposed to improve the scaffold bioactivity. The experimental results show, with the use of the two methods, the scaffolds are successfully covered by the deposition of the nano-net structure calcium phosphate coating, and they hold their three dimensional interconnected porous structures. Therefore, this kind of bioactive composite scaffold with such mechanical strength as that of woven bone should be a promising bone graft in clinical applications. PMID- 19813614 TI - [A new radiopharmaceutical for osteoarthral cartilage imaging: preparation and animals experiment of 99mTc-CS]. AB - The objective of this research work is to prepare a new and reliable radiopharmaceutical for osteoarthral cartilage imaging. Chondroitin sulfate (CS) was labeled directly with 99mTc by "pretinning" method, the labeling efficiency was 79.30% +/- 1.72% and radiochemical purity was 90.12% +/- 1l.23% after filtration by use of asepsis filter membrane. The biodistribution of 99Tc-CS was studied at 10, 30 min and 1, 2, 3, 4, 5, 6, 7, 8, 9 h respectively after caudal vein injection in normal mice,which showed that the radiolabeled product was washed out from blood quickly, and it was mainly excreted through kidney, but 99mTc-CS had distinctive characteristic of philo- cartilage. Scintigraphy of 99mTc-CS was performed on the rabbit with ostarthritis at 5 h after intravenous injection; the scintigram was clear,and the focus showed increased radiopharmaceutical uptake. The radioactivity ratio of the injured/uninjured side (T/NT) calculated over the region of interest (ROI) was 3.0 +/- 0.6 (P < 0.05). These results indicate that 99mTc-CS is an easily prepared compound with the characteristic of high philo-cartilage and it may be a new ideal cartilage imaging agent. PMID- 19813615 TI - [Bio-effects of nano-TiO2 on lungs of mice]. AB - To evaluate the acute lung toxicity of intratracheally instilled nano-TiO2 in Kunming mice, healthy adult male Kunming mice were randomly grouped by their body weight (5 mice in each group). The lungs of mice were intratracheally instilled with 1 or 10 mg/kg x bw of nano-TiO2. The control group was intratracheally instilled with the same volume of physiological saline. After 1 d, 7 d, 14 d and 28 d of exposure, the bronchoalveolar lavage fluid (BALF) and lung tissue were collected. The indices of BALF were examined. Lung tissues were assess histopathologically. The results showed that all indices of 10 mg/kg x bw groups were obviously higher than those of the control group and the group of nano-1 mg/kg x bw, respectively. Activities of lactate dehydrogenase (LDH) on the 1st, 7th, 14th and 28th day post-exposure (pe), the amounts of malodialdehyde (MDA) on the 1st, 7th and, 14th day pe and total protein (TP) on the 1st and 7th day pe as well as the amounts of leukocyte on the 1st and 7th day pe of 10 mg/kg x bw groups were significantly different as compared with controls (P < 0.05). There were no obvious changes observed in the activity of alkaline phosphatase (ALP) within groups (P > 0.05). Histopathological examination revealed that the lungs of 10 mg/kg x bw groups presented marked increase in pulmonary inflammation. Many TiO2 particles were still clearly found in the interstitium at 28 days pe. In contrast, low-dose instillation put forward a low risk potential for producing adverse effects on pulmonary health. We conclude that the inflammatory reaction gradually ceased after 28 days. Under the same experimental condition, the effect of lung injury was severer in high-dose nano-TiO2 than in low-dose nano-TiO2. PMID- 19813616 TI - [The cytocompatibility of self-assembly hydrogel from the neotype of amphiphilic peptide with neural stem cells]. AB - Neural stem cells (NSCs) cultured in vitro were implanted within three dimensional (3D) and self-assembly hydrogel from IKVAV-containing peptide. The cytocompatibility of hydrogel with NSCs was explored. Neural cells harvested from the cerebral cortex of neonatal mice were dissociated mechanically and cultivated in serum-free media, and were immunohistochemically examined for Nestin, NSE (neuron specific enolase) and GFAP (Glial fibrillary acidic protein); the self assembly hydrogel from 1 wt% amphiphilic peptide was formed with the addition of DMEM/F12 and observed under transmission electron microscope (TEM); 1 x 10(5) /ml NSCs implanted within hydrogel (3D culture system) and seeded onto the surface of coverslips covered with polylysine (two-dimensional culture system, 2D culture system) respectively were incubated without serum at 37 degrees C in 5% CO2 and 95% air. Then the cells in 3D culture system and 2D one were double-labeled with DAPI/Brdu and observed under fluorescence microscope. The Nestin-positive cells were found, and they were able to differentiate into NSE-positive neuron- like cells and GFAP-positive glial-like cells, TEM showed that the hydrogel derived from 1 wt% peptide solution was composed of nanofiber network with the fiber diameters ranging from 3 to 6 nm and the fiber length warying from 100 nm to 1.5 microm, NSCs labeled with Brdu in 2D system were thinly scattered, and those in 3D system formed many neurospheres; the positive rate in 3D was much higher than that in 2D system (P < 0.001). The self-assembly hydrogel from the amphiphilic peptide containing IKVAV sequence had good cyto-compatibility and promoted the proliferation of NSCs. PMID- 19813617 TI - [A comparative study of PTT and CT tests for coagulation evaluation of cardiovascular system external communicating devices]. AB - Based on GB/T16886.4-2003 Standard, the coagulation effects of 5 cardiovascular system external communicating devices made consist of metal and polymer were assessed using the partial thromboplastin time (PTT) and Lee-White coagulation time (CT) tests. The results indicate that PTT test is a stable and valuble method for evaluating the coagulating pathway disturbance of the devices. In line with GB/T 16886.4-2003 Standard, PTT test is a recommendable method for evaluating the external communicating devices Based on GB/T14233.2-2005 Standard, CT test is a method more liable to variation, compared with PTT test. This is due to the complex relativity in the test itself, due to all the factors of coagulating cascade, and due to the individual difference of animal. The question of how to select proper negative control for coagulation evaluation of the Cardiovascular System External Communicating Devices in clinical setting for CT test should be addressed and studied. PMID- 19813618 TI - [Research on prerequisite for using bovine hemoglobin modified by polyethylene glycol as a red blood cell substitute]. AB - This paper aims to raise concerns about the modification of bovine hemoglobin with the use of SC-mPEG as modifying reagent. The yield rates of SC-mPEG-BHB under dissimilar conditions are studied, and more important, a series of methods used to characterize the SC-mPEG-BHB mixtures are built. The best reactive system has been confirmed, i.e., at 4 degrees C, pH9; SC-mPEG and BHB are mixed according to the proportion of 60:1 and be submitted to reaction for 2 h. PMID- 19813619 TI - [Cloning and expression of extracellular region gene located in N-terminus of Leishmania Donovani]. AB - The objective of this study was to construct and express recombinant prokaryotic plasmid pET32a (+)- ast1 in E. coli BL21(DE3). Amastin gene was amplified from genomic DNA of Leishmania Donovani and its transmembran region was predicted by the methods of SOSUI and Tmpred; astl located in N-terminus of amastin gene was amplified and cloned into prokaryotic plasmid pET32a(+), which was named pET32a(+)-ast1, and then rAST1 was expressed in E. coli BL21(DE3). The results of SDS-PAGE and immunobloting assay showed that a fusion protein rAST1 (relative molecular mass about 27 kDa) was able to express in BL21. The recombinant prokaryotic plasmid pET32a(+)- ast1 was successfully constructed, and noted to be efficiently expressed in E. coli BL21(DE3). PMID- 19813620 TI - [Effect of ultrasound activating hematoporphyrin on the activities of antioxidative enzymes in mouse hepatoma 22]. AB - This investigation was made with regard to the influences of ultrasound combined with hematoporphyrin on the activities of antioxidative enzyme in ascites hepatoma 22 (H-22) tumor cells, and to a better understanding of the potential biological mechanism of sonodynamic therapy which involved the damage to cells. Combined with 100 microg/ml hematoporphyrin, high intensity focused ultrasound sonication at a frequency of 1.43 MHz and an intensity level of 2.0 W/cm2 was delivered to H-22 tumor cells for 1 min. The viability of cells was evaluated by typan-blue blue exclusion test. The intracellular reactive oxygen species (ROS) levels were determined by 2',7'-dichlorofluorescein diacetata (DCFH-DA). Enzymatic chemical methods were used to measure the activities of key antioxidative enzymes. The results indicated that the cell damage rate of ultrasound combined with hematoporphyrin was significantly higher than that of the treatment with ultrasound alone, and hematoporphyrin alone had no killing effect on H-22 cells. The level of ROS in cell suspension was significantly increased, and the key antioxidative enzyme activities were obviously decreased after treatment with the combined use of ultrasound and hematoporphyrin. We speculated that the decreased activities of key antioxidative enzymes in cells might be involved in mediating the killing effect on H22 cells in sonodynamic therapy. PMID- 19813621 TI - The Bax BH3 peptide H2-H3 promotes apoptosis by inhibiting Bcl-2's pore-forming and anti-Bax activities in the membrane. AB - Pore-formation and protein-protein interactions are considered to play critical roles in the regulation of apoptosis by Bcl-2 family proteins. During the initiation of apoptosis, the anti-apoptotic Bcl-2 and the pro-apoptotic Bax form different pores to regulate the permeability of mitochondrial outer membrane, playing their opposite functions. Overexpression of Bcl-2 has been found in various cancer cells, therefore it is gaining widespread interest to discover small molecules to compromise Bcl-2 function for anti-cancer treatment. Since Bax binds to Bcl-2's hydrophobic groove via its BH3 domain (composed of helices 2 and 3), by which their functions are inhibited each other, the H2-H3 peptide that contains the functional BH3 domain of Bax has been considered as a potential Bcl 2 antagonist. We recently reported that Bax peptide H2-H3 promotes cell death by inducing Bax-mediated cytochrome c release and by antagonizing Bcl-2's inhibitory effect on Bax. However, the mechanism of how H2-H3 inhibits the anti-apoptotic activity of Bcl-2 remains poorly understood. To address this question, we reconstituted the Bcl-2 or Bax pore-forming process in vitro. We found that H2-H3 inhibited Bcl-2's pore formation and neutralized Bcl-2's inhibitory effect on Bax pore formation in the membrane, whereas the mutant H2-H3 peptide that does not induce apoptosis in cells was shown to have no effect on Bcl-2's activities. Thus, inhibiting Bcl-2's pore-forming and anti-Bax activities in the membrane is strongly correlated with H2-H3's pro-apoptosis function in cells. PMID- 19813622 TI - [Pressure mediated effects on proliferation, apoptosis and apoptosis-associated protein of endothelial cells in the flow field in vitro]. AB - The changes of pressure in local circulation flow field and the alterations of biorheological characteristics in Endothelial cells (ECs) would follow the geometric changes of cardiovascular wall structures and would further result in distinct pathophysiological changes of endothelial cellular proliferation and vitality. This experiment is designed to observe the effects of pressure shift on ECs proliferation, apoptosis, and expression of apoptosis-associated protein, to elucidate the influences of pressure shift on the vitality of ECs, and to shed light on the dose-effect relationship concerned. By adopting flow cytometery, transmission electron microscopy, real-time RT-PCR and Western blotting, we set the levels of pressure loading ECs groups and set down the non-activated cultured ECs,single shear stress loading ECs as the control group for studies on the ultra structure alterations, on the distribution of cell cycle and the changes of proliferation and apoptosis in ECs. We also investigated the changes of the expression of Caspase-3 gene and the changing regularity of P53, Bcl-2 and Fas protein at the translation level. When ECs being exposed to decreased pressure shift (-40 cmH2O), distinct apoptosis in ECs could be observed and a pattern of duration-dependence was seen. The expressions of P53, Bcl-2 and Fas proteins are essential for regulating the genesis and process of ECs apoptosis induced by -40 cmH2O pressure. PMID- 19813623 TI - [Relationships of human laryngeal squamous cell carcinomas with the expression of VEGF-C and VEGFR-3]. AB - In order to detect the relationships of human laryngeal squamous cell carcinoma (LSCC) with the expression of Vascular Endothelial Growth Factor C (VEGF-C) and Vascular Endothelial Growth Factor Receptor-3 (VEGFR-3), we examined the level of VEGF-C and VEGFR-3 gene expression in the samples from 30 cases of normal laryngeal mucosa tissue (NLM), primary laryngeal carcinoma (PLC) and cervical lymph node (CLN) by using reverse transcription polymerase chain reaction (RT PCR). The protein levels of VEGF-C and VEGFR-3 expression were also determined by Western blotting in 10 cases of NLM, PLC, CLN,respectively. We found that in the samples from the same patient, the VEGF-C and VEGFR-3 expression level is different among normal laryngeal mucosa tissue, primary laryngeal carcinoma and cervical lymph node. In primary laryngeal carcinoma, the level of VEGF-C and VEGFR-3 expression was significantly higher in lymph node positive group than in lymph node negative group, and this difference was the associated with the histological grade of differentiation (P < 0.05). The expression levels of VEGF-C and VEGFR-3 were not associated with age, sex, site and T stage (P > 0.5). These results suggest that there is a close relationship of LSCC with VEGF-C/VEGFR-3; and VEGF-C/VEGFR-3 may have the potential for use as an indicator with prognostic significance in dealing with LSCC. PMID- 19813624 TI - [Study on cryopreservation of tissue engineered tendon by vitrification]. AB - In search of a practical method for the cryopreservation of tissue engineered tendon (TET) by vitrification, we adopted 3 kinds of different cryoprotective agents (CPA)(21% DMSO, DP6 and VS55) in studying the freeze-stored effect of different CPA. The cellular morphology and post-thaw viability of the TET were examined by scanning electron microscopy (SEM), flow cytometry, and confocal laser microscopy (CLM). The results showed that there existed statistically significant difference in respect to the post-thaw viability between 21% DMSO and DP6, VS55; The cells specially adhered to the surface of scaffold both before or after cryopreservation by use of 21% DMSO. It was suggested that 21% DMSO as a CPA for TET cryopreservation was better than DP6 and VS55 in the current study. PMID- 19813625 TI - [Effect of body mass index on bone mass of lumbar spine of male patients with ostealgia and arthralgia but without osteoporosis]. AB - For the purpose of identifying the effect of body mass index on bone mineral density, an investigation was made in regard to the relationship between body mass index (BMI), bone mineral density (BMD) and bone mineral content (BMC) of lumbar vertebrals of men without osteoporosis. 838 male patients were diagnosed and treated in West China Hospital of Sichuan University because of ostealgia and arthralgia from Aug. 2003 to Dec. 2005. They were examined by dual-energy X-ray absorptiometry, their L2-L4 bone mineral density being < -2.5SD. Then they were graded by body mass index (Kg/m2) [1st group BMI > or =23, 454 cases (Group A BMI > or = 29, 46 cases; Group B 25 < or = BMI < 28.9, 201 cases; Group C 23 < or = BMI < 24, 9193 cases); 2nd group 18.5 < BMI < 22.9, 311 cases; 3rd group BMI < or = 18.5, 68 cases]. The relationships between BMI and BMD of lumbar spine were analyzed using SPSS 13.0. The results revealed their positions in series: (1) Among Groups 1-3, BMD, average BMD and aBMC of L2-L4 being 3rd group > 2nd group > 1st group, the differencs are statistically significant (P < 0.05 or P < 0.01); BMC, aBMC of L3 and L4 and aBMC of L2 being 3rd group > 2nd group > 2nd group, the differences are significant (P < 0.01); besides BMC of L2 and T-score,the differencs among three groups are not statistically significant (P > 0.05); (2) Among the 1st group, BMD and average BMD of L2-L4 being Group C > Group B > Group A,the differencs are significant (P < 0.05 or P < 0.01); BMC, aBMC of L and I being Group C > Group B > Group A,the differences among the three groups are significant (P < 0.05 or P < 0.01); BMC, aBMC of L3 and L4 and aBMC of L2 being Group C > Group B > Group A, the differences are significant, besides BMC of L2 and T-score,the differences among three groups are not significant (P > 0.05), (3) BMI correlated negatively (P < 0.01, r = -0.189) to the average BMD of lumbar spine. The average BMD of lumbar spine decreases when BMI increases in males without osteoporosis. When obesity is more obvious, the decrease in average BMD of lumbar spine bone is more evidently decreased. The decrease of BMD, BMC and total BMC in L3 and L may be greater than that in L2. PMID- 19813626 TI - [Relationship between apolipoprotein E polymorphism and cognitive function in patients with primary hypertension]. AB - To explore the relationship between apolipoprotein E polymorphism and cognitive function in primary hypertension patients, we collected 200 Chinese primary hypertensive patients. Blood pressure (BP), heart rate (HR), height, body weight, waistline, hip circumference were measured. The Mini Mental State Examination (MMSE) was applied to test the cognitive function and compute score. Full automatic bio-chemistry analyzer was used to determine total cholesterol (TC) and triglyeride (TG) and fasting glucose. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RELP) was used for the analysis of the apolipoprotein E polymorphism. We found that in primary hypertension patients, the genotype frequency of epsilon3/4 and epsilon4/4 were significantly higher in the cognitive impairment group than that in the cognitive normal group. The allele frequency of e4 is obviously higher in the cognitive impairment group than that in the cognitive normal group. Age and epsilon4/4 genetype were positively correlated with hypertensive-cognitive impairment, while cultural level was negtively correlated with it. ApoEepsilon4 allele and age might be risk factors for the cognitive impairment in hypertensive patients. The epsilon4 homozygote (epsilon4/4) might be an important influencing factor for the progression of cognitive impairment. PMID- 19813627 TI - [Segmentation of multiple sclerosis lesions based on Markov random fields model for MR images]. AB - Multiple sclerosis (MS) is an inflammatory demyelinating disease that would damage central nervous system. There is a growing attention to the segmentation algorithms of MS Lesions. An MRF-based algorithm for MS lesions segmentation of T2-weighted MR brain images is developed by utilizing the morphological characteristics of MS lesion tissues. The regions circumscribed by white matter are extracted at first by MRF-based segmentation and region growing methods; the abstracted regions are then segmented again using MRF-based algorithm. The segmented MS lesions of both simulated and clinical T2-weighted MR brain images are presented in the current work. The testing results show that the proposed algorithm is robust and accurate enough for clinical use. PMID- 19813628 TI - [Establishment of database with standard 3D tooth crowns based on 3DS MAX]. AB - The database with standard 3D tooth crowns has laid the groundwork for dental CAD/CAM system. In this paper, we design the standard tooth crowns in 3DS MAX 9.0 and create a database with these models successfully. Firstly, some key lines are collected from standard tooth pictures. Then we use 3DS MAX 9.0 to design the digital tooth model based on these lines. During the design process, it is important to refer to the standard plaster tooth model. After some tests, the standard tooth models designed with this method are accurate and adaptable; furthermore, it is very easy to perform some operations on the models such as deforming and translating. This method provides a new idea to build the database with standard 3D tooth crowns and a basis for dental CAD/CAM system. PMID- 19813629 TI - [A study of sleep stage classification based on permutation entropy for electroencephalogram]. AB - This paper presents a new method for automatic sleep stage classification which is based on the EEG permutation entropy. The EEG permutation entropy has notable distinction in each stage of sleep and manifests the trend of regular transforming. So it can be used as features of sleep EEG in each stage. Nearest neighbor is employed as the pattern recognition method to classify the stages of sleep. Experiments are conducted on 750 sleep EEG samples and the mean identification rate can be up to 79.6%. PMID- 19813630 TI - [Feature detection of tissue lesion in HIFU based on SOM]. AB - Noninvasive detection of tissue lesion is an important aided way in the process of high intensity focused ultrasound (HIFU) treatment. A new method is proposed for noninvasive detection of tissue lesion based on self-organizing maps (SOM) network. Tissue lesion, created by HIFU exposures, inevitably leads to change in B-mode image, since some new interfaces will appear in tissue. The difference-sub image near focus of HIFU, where the tissue lesion appears, is used. Feature of lesion in difference-sub-image is extracted using the method based on SOM network. Experimental results show that the performance of the method proposed in this paper is better than that of others, such as the methods based on average gray, energy and standard deviation of difference-sub-image. PMID- 19813631 TI - [Design and development of the multi-dimensional ultrasonic reconstruction and visualization system]. AB - The design and development of multi-dimensional ultrasonic reconstruction and visualization system (MURVS) have been described in the present paper. This system is basically composed of four modules: the data input/output module, image segmentation and arrangement module, multi-dimensional reconstruction module, and the dynamic visualization module. At first, some algorithms used in the system are introduced by the authors, including the AVI segmentation algorithm, three dimensional interpolation algorithm, and volume rendering algorithms. Then the key questions of techniques to be discussed are: how to design the main modules, how to solve the dynamic visualization problem, and how to implement the system. The experiments indicate that MURVS is able to reconstruct all three-dimensional data fields in one cardiac cycle of a patient within 4 seconds, and dynamically display the motion of the heart. It allows the medical professionals to select different parameters when observing the reconstructed results. This is very helpful for medical professionals to reach more accurate diagnoses of their patients' diseases. PMID- 19813632 TI - [Research on piezoelectric protein sensor array for rapid detecting HIV(1+2) antibody]. AB - To research piezoelectric immunosensor array for rapid detecting HIV(1+2), piezoelectric immunosensor array matrix was designed. HIV(1+2)C1 antigen was immobilized onto the silver electrodes of quartz-crystal microbalance, which was modified with adsorption and cross-linked method. In the clean air flow and monitoring environment, standard quality control and clinical serum sample were detected. The linear range for the measurement of HIV(1+2) was 0.01-0.2 NCU/ml. The sensitivity, specificity and accuracy of HIV(1+2) piezoelectric protein sensor array were 91.7%, 93.3% and 92.7% respectively. PMID- 19813633 TI - [Detection of endpoint for segmentation between consonants and vowels in aphasia rehabilitation software based on artificial intelligence scheduling]. AB - For the purpose of improving the efficiency of aphasia rehabilitation training, artificial intelligence-scheduling function is added in the aphasia rehabilitation software, and the software's performance is improved. With the characteristics of aphasia patient's voice as well as with the need of artificial intelligence-scheduling functions under consideration, the present authors have designed a set of endpoint detection algorithm. It determines the reference endpoints, then extracts every word and ensures the reasonable segmentation points between consonants and vowels, using the reference endpoints. The results of experiments show that the algorithm is able to attain the objects of detection at a higher accuracy rate. Therefore, it is applicable to the detection of endpoint on aphasia-patient's voice. PMID- 19813634 TI - [A review of studies on a subset of rapidly self-renewing marrow stromal cells]. AB - A series of studies have demonstrated that bone marrow mesenchymal stem cells (MSCs) are attractive candidates for cell and gene therapies, because they are readily obtained and multipotentially differentiated. Then homogeneous MSC cultures in vitro with more rapidly self-renewing ability and multipotential differentiation will accelerate and improve their progress in clinical application. Colter et al. found that early colonies contain a third kind of cells very small round cells that rapidly self-renew, besides spindle-shaped cells and large flat cells, called RS cells. RS cells are characterized by their extremely small size, rapid rate of replication, and enhanced potential for multilineage differentiation. Moreover, they can be distinguished from more mature cells in the same cultures by a series of surface epitopes and expressed proteins. Therefore, the results raise the possibility that RS cells may have the greatest potential for long-term engraftment and differentiation in vivo. PMID- 19813635 TI - [The progress of research on the influencing factor of nuclear transfer]. AB - Nuclear transfer (NT) is a new cloning technology developed in recent years. NT methods consist of electrofusion, NT mediated by polyethylene glycol (PEG) and microinjection. The success of somatic nuclear transfer depends on the source of donor nucleus, developmental stage of recipient cytoplasts, cell cycle synchrony of donor nucleus. Different methods of harvesting cells have effect on the efficiency of NT. The somatic nucleus will be reprogrammed after NT and will restore a totipotent state in order to undergo development. PMID- 19813636 TI - [Progresses in recombinant human serum albumin]. AB - Human serum albumin (HSA) is an important biological product in clinical pharmacy practice at present. Its main clinical use is in maintaining colloid osmotic pressure and increasing circulating plasma volume so as to cure hemorrhagic shock, burn, cancer, hypercytosis, hypoalbuminosis, etc. HSA is isolated by fractionating human plasma, which entails possible contamination by viruses or prions. Recombinant human serum albumin (rHSA) has been successfully produced by biological engineering. The structural and functional properties of rHSA are similar to those of plasma derived human serum albumin (pdHSA). Preclinical and clinical trials have confirmed the safety and efficacy of using this rHSA preparation in the treatment of certain diseases. PMID- 19813637 TI - [Advancement of researches on relationship between hepatitis B virus and severe hepatitis B]. AB - Approximately 1% of patients infected with hepatitis B virus (HBV) develops severe hepatitis B, which is characterized by rapid destruction of liver parenchyma. The case fatality rates of HBV-induced severe hepatitis B in China range between 60% and 80%. In other words, more than half of the patients with severe hepatitis B ultimately require liver transplantation or die of liver failure. The pathogenesis of severe hepatitis B is not well understood. Previous studies have indicated that complicated interaction between viral factors and host immune response may be involved. This article tries to review the advancement of researches on the relationship between HBV and severe hepatitis B. PMID- 19813638 TI - [Progress in detection, analysis and application of new techniques to monitoring bowel sounds]. AB - The detection and analysis of bowel sounds are of importance to the diagnosis and treatment of gastrointestinal diseases. In this paper are summarized the recent advances in the detection and analysis as well as in the application of new techniques to monitoring bowel sounds. Several methods, including spectrum analysis, adaptive filtering, wavelet transform, principal component analysis, and artificial neutral network are mainly introduced. All of them are promising for feature extraction and automatic classification of bowel sounds. PMID- 19813639 TI - [Research progress of stochastic resonance in neural models]. AB - In nonlinear systems, noise can improve the responses of the systems with appropriate noise intensity. This phenomenon is called stochastic resonance. Biological neural systems are noisy and stochastic resonance has been found in them experimentally and theoretically. Now many researches focus on the signal transmission and processing in neural models. So this paper introduces the researches of stochastic resonance in noisy neural models. Then the recent research achievement and progress are reviewed in the following three aspects: noise; the development of stochastic resonance; and neural network. At last, the foreground of the study is discussed. PMID- 19813640 TI - [Advances in noninvasive measurement of ankle-brachial index]. AB - The key of Ankle-brachial index (ABI) measurement is the synchronous measurement of four limbs' systolic blood pressures. In this paper is analyzed the inadequacy of the modern blood pressure measurement technologies used in the ABI measurement process. Special emphasis is laid on the principles and characteristics of the double-layer cuffs technology. The research orientation, the existing problems, and the way toward improvement are discussed. PMID- 19813641 TI - [The study and clinical application of absorbable hemostatic agent]. AB - This paper reviews the mechanisms and properties of different absorbable hemostatic agents. The development tendency of absorbable hemostatic agent is forecasted. Its' qualities of being useful to surgeons are introduced and are embodied in surgeon's comprehending of the hemostatic characteristic of absorbable hemostatic agents as well as in their clinical use of such agents accurately. At the same time, the researchers in pursuit of the medical materials can work with reference to the data herein presented. PMID- 19813642 TI - [Antitumor drug delivery system: the progress of researches on block copolymer micelle and its compositive materials]. AB - In antitumor drug delivery system, the block copolymer micelle is a sort of new colloidal state drug carrier in recent years, and it has lots of remarkable properties such as high stability in vitro and in vivo, favourable biocompatibility, controlling of drug release, and targeting to tumor tissue. At present, a great deal of researches have been madeon its compositive materials. Except commonly used micelle materials, the new developmental intelligent micelle materials have been becoming the hotspots of researches on block copolymer micelle. This review is presented with emphasis on the block copolymer micelle and its compositive materials. PMID- 19813643 TI - [A half-colored revolution]. PMID- 19813644 TI - [Nineteenth World Congress of Gerontology and Geriatrics. How can one live a better old age?]. PMID- 19813645 TI - [The care, the nurses and the families in geriatrics]. PMID- 19813646 TI - [Suicide in the aged, a hidden reality]. PMID- 19813647 TI - [Seasonal entertainment, a different approach for entering an institution]. PMID- 19813648 TI - [The fabric of Saint Morpheus, a partner in taking care of sleep problems]. PMID- 19813649 TI - [Nursing care to help along the way]. PMID- 19813650 TI - [A packet of good treatment for considering practices in geriatrics]. PMID- 19813651 TI - [Cardiology in the aged. The heart in all its conditions]. PMID- 19813652 TI - [Cardiology in the aged, the state of positions and perspectives]. PMID- 19813653 TI - [Arterial hypertension and orthostatic hypotension in the aged]. PMID- 19813654 TI - [Heart insufficiency in the aged]. PMID- 19813655 TI - [Coronary diseases in the aged, the role of the interventional therapeutics of revascularization]. PMID- 19813656 TI - [The position of holter monitoring and electrocardiography, between emotion and resignation]. PMID- 19813657 TI - [Venous thromboembolism; specific geriatric considerations in care]. PMID- 19813658 TI - [11/12 Massage-kinesthesia therapy]. PMID- 19813659 TI - [Nurses, usage and families. 8/8--regulatory framework of organ donation]. PMID- 19813660 TI - [5/6 Care and treatment of the aged]. PMID- 19813661 TI - [Influenza pandemic, we must remain vigilant!]. PMID- 19813662 TI - [Parenthood from proof to practice]. PMID- 19813663 TI - [We should prepare ourselves for the future of therapeutic education of the patient]. PMID- 19813664 TI - [Setting limits without producing violence in adolescents]. PMID- 19813665 TI - [Cross-breeding and diversity, an opportunity]. PMID- 19813666 TI - [Parenthood in exile]. PMID- 19813667 TI - [A clinic for half-castes for children of immigrants]. PMID- 19813668 TI - [Selective inability to speak, a expression of division between two cultures]. PMID- 19813669 TI - [Adolescents from here versus from other places]. PMID- 19813670 TI - [Favoring the prevention of postural disorders in premature births]. PMID- 19813671 TI - [Music and noises attending neonatology]. PMID- 19813672 TI - [6/12. Convulsions in childhood]. PMID- 19813673 TI - [6/7. Resuscitation of the newborn and at the end of life]. PMID- 19813674 TI - Laparoscopic repair of incisional hernia: a valid option. PMID- 19813675 TI - Swine flu--a new threat to community. PMID- 19813676 TI - Association of Helicobacter pylori infection with idiopathic thrombocytopenic purpura. AB - OBJECTIVE: To determine the association of Helicobacter pylori infection in patients presenting with idiopathic thrombocytopenic purpura (ITP). METHODS: From March 2007 to March 2008, thirty adult patients with ITP and 30 age and sex matched healthy controls were investigated for the presence of H. pylori infection by Helicobacter pylori stool antigen (HpSA) an enzyme immunoassay (EIA) based method. The criteria for presence of H. pylori infection was a positive stool antigen test. RESULTS: H. pylori infection was found in 19 out of 30 patients with ITP (63.3%) which is well above the frequency of 13 out of 30 (43.3%) in controls. Calculated odds ratio was 2.25 which shows significant association of H. pylori infection with ITP. CONCLUSION: The study confirms the existence of an association between H. pylori infection and ITP. Therefore the screening for H. pylori infection and an attempt to eradicate bacterium in positive cases seems appropriate in patients with ITP at diagnosis. PMID- 19813677 TI - Determination of complication rate of PICC lines in oncological patients. AB - OBJECTIVE: To determine the complication rate of (PICCs) peripherally inserted central catheters in cancer patients with a 1 year prospective cohort study. METHODS: All PICCs inserted in adult cancer patients in Radiology Department of The Aga Khan University Hospital were followed prospectively till removed or patient expired and pattern of complications noted. RESULTS: One hundred and fourty six PICCs were inserted over a period of 1 year and followed for a total of 3329 catheter-days; median placement, 14 days: range 3-218 days. Of these 67 (32.8%) PICCs were complicated and removed earlier, for a rate of 14.4/1000 PICC days. Patients with haematologic malignancies were more likely to have complications as compared to those with solid tissue malignancies. CONCLUSIONS: Despite a significant complication rate, PICCs are a relatively safe and cost effective method of establishing central venous access. PMID- 19813678 TI - Comparison of non cardiac chest pain (NCCP) and acute coronary syndrome (ACS) patients presenting to a tertiary care center. AB - OBJECTIVE: To compare clinical characteristics of patients admitted with chest pain with those who had Acute Coronary Syndrome (ACS) and Non Cardiac Chest Pain (NCCP) presenting at a tertiary care center. METHODS: An analytical crossectional study was done. All patients presenting to the emergency with acute chest pain of age more than 18 years who had electrocardiography done and had an initial serum Troponin I (Trop I) measured were included. Patients were assigned to ACS groups or NCCP group after cardiac workup. Those who did not have cardiac workup were excluded from the study. RESULTS: We enrolled a total of 202 patients. After workup 45.94% were placed in the ACS group and 34.5% in NCCP group. On comparison of the baseline characteristics of the ACS and NCCP group, there was significant difference in age (p <0.001) and Diabetes Mellitus (p <0.002). Comparison of clinical characteristics, showed a significant difference in the character of chest pain (p <0.001), electrocardiogram (p <0.001), -ve Troponin (Trop I) value (p <0.001), pulse (p <0.02) and presence of Pulmonary edema (p <0.006). Non ST elevation myocardial infarction (NSTEMI) (46.5%)was the most common diagnosis in the ACS group and Muscular pain (27.60%) was the most common diagnosis in the NCCP group. CONCLUSION: Patients with NCCP were younger, majority were non- diabetics, had slower pulse, more atypical/non-cardiac chest pain, had more normal ECG at base line as compared to patients with ACS. Muscular chest pain was the most common diagnosis in NCCP and NSTEMI in ACS group. PMID- 19813679 TI - Risk factors and clinical outcome of placental abruption: a retrospective analysis. AB - OBJECTIVE: To determine frequency, obstetrical risk factors and the subsequent feto-maternal outcome in women suffering from placental abruption. METHODS: A retrospective case series study was carried out in the Department of Obstetrics and Gynaecology Unit One, Liaquat University Hospital Hyderabad, Pakistan from 1st January 2006 to 31st December 2006. All women with the diagnosis of placental abruption having more than 24 weeks gestation were included in the study. RESULTS: Of the 2224 delivered women 106 (4.7%) had placental abruption. All of the 106 women were unbooked, with 67 (63%) in the age group 20-35 years, 68 (64%) belonged to rural areas. 98 (92%) patients were multiparous and 57 (54%) were preterm. The commonest medical disorders observed were anaemia in 84 (79%), Diabetes Mellitus in 8 (8%) and gestational hypertension in 8 (8%) patients. There were five maternal deaths, showing case fatality rate of 5%. The foetal prognosis was characterized by low birth weight seen in 74 (70%), low apgar score in 30 (28%) and high still birth rate in 54 (51%), constituting perinatal mortality rate of 25.62/1000 deliveries. CONCLUSION: Abruptio placentae is associated with adverse maternal and foetal outcome. Multiparity, un-booked status, rural residence and maternal anaemia are important risk factors. PMID- 19813680 TI - Gastrointestinal somatization in males and females with depressive disorder. AB - OBJECTIVE: To assess gastrointestinal symptoms among depressed patients presenting in an outpatient psychiatric clinic. METHODS: All consecutive and consenting cases of depressive disorder presenting to the clinic from Jan' 02 to Dec' 05 were inducted in the study. A descriptive study was carried out in a clinical setting at the Neuro-Spinal Medical Institute, Karachi, Pakistan. The assessment was based on detailed history, physical examination, mental state examination and filling of semi structured proforma designed for this purpose. For diagnostic purpose WHO classification ICD-10 criteria was used. RESULTS: A total of 1165 (Male: 56%, Female: 43.2%) patients were assessed. The age ranged from 6 to 90 years for male and 5 to 80 for females with p-value of 0.001. Majority were married. Their educational status revealed that 20.4% of males and 45.1% females were illiterate. The most frequently reported gastrointestinal symptom was decreased appetite (67.7%) followed by constipation (57.7%) and diarrhoea (42.3%). Other reported symptoms included abdominal pain, vomiting, gas, indigestion and nausea in order of frequency. In males 13.7% and 12.7% females had suffered for > 10 years before seeking psychiatric consultation. Surprisingly, about 26.3% of the patients had previously pursued alternative modes of treatment (homeopathic, faith healers and hakeems). CONCLUSION: Somatic symptoms manifesting as gastrointestinal symptoms are common in patients with depressive disorder. Awareness about such an association is essential and will be useful for primary care physicians and gastroenterologists for establishing early diagnosis and management, thus avoiding unnecessary investigations in patients with poor resources. PMID- 19813681 TI - Informed consent in elective surgical procedures: "what do the patients think"? AB - OBJECTIVE: To analyze the patients' perspective of the process of informed consent in a tertiary care hospital setting. METHODS: A cross sectional survey was carried out at a tertiary care teaching hospital Islamabad and 200 patients were included who responded to the questionnaire. The study extended from March to May 2008. Patients were interviewed by trained personnel. Patients undergoing emergency surgery, minors and medical professionals were excluded form the study. Data was entered and frequencies and percentages were analyzed on SPSS 14. RESULTS: A total of 98% of patients admitted that informed consent was taken, however, only 81 (40.5%) patients understood the information provided. Patients reporting that their decision to proceed with surgery was actually influenced by other people including the treating doctor numbered 112 (56%). Only 58 (29%) signed their own consent form, the rest of them were signed by relatives. Only 96 (48%) patients admitted having been informed about possible complications of surgery as against 156 (78%) who were informed of sequelae of not having the surgery done. Signatures taken by the paramedics were 138 (69%). Despite a poor understanding of the process of informed consent, 187 (93.5%) still felt satisfied with the process of informed consent. CONCLUSION: There is a lack of proper understanding by the population and inadequate system of informed consent in our setting. However, majority of the people still feel satisfied with the information provided. There is a unique family system where decision-making is shared within the family. PMID- 19813682 TI - Precipitating factors of hepatic encephalopathy at a tertiary care hospital Jamshoro, Hyderabad. AB - OBJECTIVE: To determine the precipitating factors of hepatic encephalopathy (HE) in patients with liver cirrhosis at Liaquat University Hospital Hyderabad/Jamshoro. METHODS: This hospital based descriptive study was conducted from April 2007 to September 2007. All the patients who were more than 12 years of age and were diagnosed as hepatic encephalopathy were studied. During this period, 87 patients of hepatic encephalopathy were admitted. All patients were carefully examined, relevant investigations were performed and data was collected through pre-designed proforma. RESULTS: Male patients were 65 (75%), above 40 years of age 58 (67%), belonging to interior/periphery of Sindh 54 (62%), in grade IV of hepatic encephalopathy 70 (80%) and Anti-HCV positive were 52 (60%). The most common precipitating factors detected were infection 58 (67%), constipation 43 (49%) and gastrointestinal bleeding 39 (45%). Out of 87 patients, 68 had increased total leucocytes count, 09 patients had hypokalaemia, 24 patients, hyponatraemia, 64 hypoalbuminaemia and 54 patients had a disturbed coagulation profile. Fifty nine patients recovered and were discharged while 20 patients expired. Majority of expired patients had Child-Pugh score 10-15 and were in grade IV of hepatic encephalopathy. CONCLUSION: The study concluded that there were different factors which play a key role in hepatic encephalopathy. In these factors, infection was the most common. PMID- 19813683 TI - Knowledge and practices regarding foot care in diabetic patients visiting diabetic clinic in Jinnah Hospital, Lahore. AB - OBJECTIVE: To assess the knowledge and practices among the diabetic patients regarding foot care. METHODS: In this cross sectional study, by using non probability convenience sampling, 150 diabetic respondents fulfilling the inclusion criteria were included in the study. Their knowledge and practices regarding foot care were assessed by a pre-tested questionnaire and classified as good, satisfactory and poor depending upon the score. Fifteen questions each were asked regarding knowledge and practices of foot care. Each question was assigned one mark. If score was more than 70% (11-15), it was regarded as good, if score was 50-70% (8-10) it was regarded as satisfactory and if score less than 50% (<8) it was regarded as poor both for knowledge and practice for foot care. RESULTS: The mean age of the respondents was 48 +/- 10.8 years. About 29.3% respondents had good knowledge, 40% had satisfactory knowledge and 30.7% had poor knowledge about foot care. Whereas only 14% respondents had good practices for foot care, 54% had satisfactory practices and 32% had poor practices. Education of the respondents had significant statistical association with knowledge (p value<0.001) and practices (p-value <0.001) regarding foot care. Sex and income per capita had shown no significant statistical association with knowledge and practices regarding foot care. CONCLUSION: About one third of diabetic patients had poor knowledge about foot care and only very few patients had good practices for foot care. Literacy has significant association with the knowledge and practices related to foot care in diabetic patients. PMID- 19813684 TI - Isolated oligohydramnios is not an indicator for adverse perinatal outcome. AB - OBJECTIVES: To assess the impact of isolated oligohydramnios on Perinatal outcomes. METHODS: A prospective Cohort Study was conducted at the department of Obstetrics and Gynaecology, The Aga Khan University Hospital, Karachi, Pakistan. Pregnant women with isolated oligohydramnios were recruited for the study after obtaining informed consent. The mode of delivery and perinatal outcome were compared with women having normal amniotic fluid. RESULTS: Between 1st May 2005 and 30th December 2005 a total of 421 women were included in this study. Of these 421 women, 71 were exposed and 350 were unexposed. When compared to the unexposed group women with oligohydramnios had significantly lower birth weight babies and were delivered at a significantly earlier gestational age. However there was no statistical difference in the APGAR scores at birth and NICU admissions between the two groups. The number of inductions and caesareans done for foetal reasons were significantly higher in the exposed group. CONCLUSION: Isolated oligohydramnios is not associated with adverse perinatal outcomes. However, it increases the risk for labour induction and Caesarean section. PMID- 19813685 TI - Treatment outcome of multi-drug resistant tuberculosis in a tertiary care hospital in Karachi. AB - OBJECTIVE: To assess the outcomes of pulmonary multidrug-resistant tuberculosis (MDR-TB) patients treated at Ojha Institute of Chest Diseases (OICD), a reference hospital for TB in Karachi, Pakistan. METHODS: Clinical study for the period 1996 2006, with follow-up until June 2007 was performed. All the culture and sensitivity proven cases of MDR pulmonary TB were initially admitted for 3-6 months till the sputum converted negative. Treatment regimen was decided on individual basis, and included 4-6 drugs. Supervised treatment was given to all patients during the hospitalization. After discharge from the hospital, patients were followed at monthly interval at the outpatient department of OICD for 18 months total. RESULTS: Five hundred and seventy nine adult patients (59.93% male) with mean age of 32.44 +/- 12.63 years were studied. All patients had a history of treatment with first line anti-tuberculosis drugs. Treatment was successful in 227 (39.2%). The mortality rate was 27(4.6%) during hospitalization. During admission 83(14.3%) left treatment and 239 (41.2%) were lost to follow-up during treatment. Treatment failure was observed in three patients. CONCLUSION: The treatment success rate in this study is satisfactory but high default rate is a challenge in the management of MDR tuberculosis. PMID- 19813686 TI - Is serum total IgE levels a good predictor of allergies in children? AB - OBJECTIVE: To study the role of Serum Total IgE levels as a marker of allergy and to see its association with different host and environmental factors, its association with different systemic allergies and with the increased allergic systemic involvement. METHODS: A cross sectional comparative study was conducted at Liaquat National Hospital Paediatric Medicine Unit and Outpatient Clinic, from December 2007 to October 2008. Two hundred and fifty eight children in the age group 6 months to 12 years meeting the inclusion criteria, were enrolled for the study. Complete blood count and serum total IgE levels were done in all patients. Data was collected and tabulated. Chi-square was applied to test the association of serum Total IgE levels with different variables using SPSS and p-value of <0.05 was taken as statistically significant. RESULTS: Out of 258 patients 166(64.37%) had raised serum total IgE levels (108 males and 92 females). Among host factors there was no significant association of serum total IgE levels with age, sex, height of the patient, positive family history of allergy or age at which symptoms developed (p =or> 0.05), though the correlation of serum total IgE levels was seen with weight of the patient (p < 0.049), early weaning (p < 0.01) and bottle feeding (p < 0.01) in children. Among environmental factors serum total IgE levels had a strong association with exposure to passive smoking (p < 0.01), pollen (p < 0.01), cold (p < 0.01), and pets (p < 0.01). Blood eosinophilia was another factor that had positive association with allergy and raised serum total IgE levels (p < 0.01). However association of serum IgE levels was not statistically significant for any systemic allergies, skin allergy (0.608), food allergy (p < 0.210), allergic sinusitis (p < 0.113), allergic rhinitis (p < 0.358), allergic conjunctivitis (p < 0.507) except for bronchial asthma (p =or<0.01) where serum total IgE levels was also associated with severity of bronchial asthma (p < 0.01). Interestingly there was no significant relationship of increased number of systemic allergies with serum IgE levels. CONCLUSION: Serum total IgE level is a good predictor of allergy in children. It is influenced by early weaning, early bottle feeding, exposure to passive smoking, pollen, cold, and pets and is associated with blood eosinophilia. Also serum total IgE level is a strong predictor of allergy in asthmatic children. PMID- 19813687 TI - CT guided transthoracic catheter drainage of intrapulmonary abscess. AB - OBJECTIVE: To determine the efficacy of CT- guided transthoracic catheter drainage of intrapulmonary abscess considering success rate versus complications. METHODS: This prospective study was carried out at radiology department of Al Noor Specialist Hospital, Makkah, Saudi Arabia, from 1.1.2003 to 31.12.2005. Nineteen patients were selected for CT guided percutaneous drainage. Under CT guidance catheter placement was carried out using Seldinger technique. RESULTS: Nineteen patients with lung abscess were selected for the percutaneous CT guided drainage. Eight (42.105%) patients encountered no complications and lung abscess completely resolved with no residual cavity. Five (26.31%) patients developed pneumothorax, which was the most common complication of this study. These patients were kept under observation and followed-up by chest X-rays. Three (15.78%) had mild pneumothorax, which resolved and needed no further management, while two (10.52%) patients developed moderate pneumothorax and chest tube was inserted. Two (10.52%) patients developed mild haemoptysis which resolved within two hours, hence, no further management was required. Two (10.52%) patients had residual cavity and surgery was performed. Congenital cystic adenomatoid malformation (CCAM) was found in both cases. Two patients out of nineteen patients (10.52%) developed bronchopleural fistula and were operated. No mortality occurred during or after the procedure. CONCLUSION: CT allows optimal placement of catheter and hence enables safe and effective percutaneous evacuation of lung abscess. The morbidity and mortality of patients with percutaneous catheter drainage is lower than with surgical resection. Hence, CT guided drainage should be considered the first therapeutic choice in most patients of lung abscess who do not respond to medical therapy. PMID- 19813688 TI - Post tonsillectomy haemorrhage: "is day care surgery safe"? AB - OBJECTIVE: To compare the frequency of haemorrhage after tonsillectomy as a day care procedure versus tonsillectomy as a one day procedure. METHODS: A comparative study was conducted at the Department of ENT and Head and Neck Surgery, Shifa International Hospital, Islamabad from January 2003 to December 2007. The sample consisted of 400 patients, divided into two groups, One group was kept in the hospital for eight hours and was labeled as the daycare group while other group was kept admitted for 23 hours stay (overnight) and was labeled as the one day group. Patients with nasal pathology, bleeding diathesis and less than 4 years of age were excluded. Patients were followed up post operatively on 7th and 14th day and were inquired about any bleeding episode from the mouth during stay at home. The data was analysed at SPSS 13. RESULTS: Post tonsillectomy haemorrhage was seen in 10 patients (2.5%), 4 (2%) occurred in day care group while 6 (3%) in one day group. All were secondary haemorrhages. Chi square test showed no significant difference in haemorrhages between the two groups. CONCLUSIONS: Tonsillectomy can safely be done as a daycare procedure as there is no significant difference of post operative haemorrhage between daycare and one day admission. PMID- 19813689 TI - National Health Accounts: lessons for Pakistan. PMID- 19813690 TI - Myotonic dystrophy and pregnancy. AB - Myotonic dystrophy is the most common neuromuscular disease in adults with a prevalence of 2.4-5.5 per 100,000. Here we describe two cases of DM and discuss their obstetric complications. Our first case concerns a 39 year old multipara whose pregnancies were complicated by recurrent abdominal pain, polyhydramnios and post partum haemorrhage which was attributed to DM. In our second case we discuss the management of a 27 year old woman with dichorionic, di-amniotic twins. Chorionic Villous Sampling at 11 weeks revealed one of the fetuses, a male; to be afflicted by DM. Selective termination of the affected twin was performed. Unfortunately, she developed severe oligohydramnios and chronic liquor leak. This resulted in the intra-uterine death of the second twin 5 days later. Our cases highlight the importance of prenatal diagnosis and prompt genetic counselling. A multidisciplinary team approach is required in the management of such high risk cases. PMID- 19813691 TI - Iatrogenic superior vena caval syndrome. AB - The superior vena caval (SVC) syndrome is a common oncological emergency requiring the quick initiation of appropriate therapy. However, it may also result from a medical procedure e.g. central catheter or temporary pacing wire insertion, with symptoms usually developing acutely and dramatically. If symptoms persist despite removal of the offending device, chemotherapy and radiotherapy are obviously precluded. Alternative treatment modalities include thrombolysis, thrombectomy devices, stents, and surgery. Clinically covert thrombosis is not uncommon, and as interventions and invasive procedures requiring central venous cannulations become commonplace, this iatrogenic complications will inevitably occur more often. Even the use of ultrasound guided insertion does not avoid catheter related obstruction. A case of an iatrogenc haemodialysis catheter related SVCS is presented and the aetiopathogensis, signs and symptoms, diagnosis, and management are discussed. PMID- 19813692 TI - Sirenomelia, the Mermaid syndrome: case report and a brief review of literature. AB - Sirenomelia, the Mermaid Syndrome is a rare and lethal congenital anomaly with an incidence of one in 60,000 to 70,000 pregnancies. Sirenomelia is characterized by complete fusion of the lower limbs, commonly associated with renal agenesis, absent external genitalia and other gastrointestinal defects. Another pathognomonic finding is the presence of single umbilical, persistent vitelline artery which is the chief distinguishing anatomic finding from Caudal Regression Syndrome. We report a case of termination of pregnancy done on the basis of ultrasound diagnosis of bilateral renal agenesis with no liquor volume. The foetus was identified to have characteristic features of Sirenomelia at the time of termination. PMID- 19813693 TI - Brachyspira species blood stream infection. AB - The spirochetes inhabiting the large intestines of humans and animals consist of a diverse group of related organisms. Intestinal spirochetosis caused by Brachyspira pilosicoli is a newly recognized enteric disease of human being and animals manifest as mild colitis and diarrhoea. Few cases have been reported of spirochetaemia especially in critically ill and immunocomprised patients. This is another case of spirochetaemia in a patient diagnosed as a case of chronic liver disease secondary to hepatitis C virus, presented in emergency room with history of fever, abdominal pain and slightly altered mental state. Brachyspira pilosicoli was isolated in anaerobic blood culture bottle. PMID- 19813694 TI - Terrorism and mental health: the issue of psychological fragility. PMID- 19813695 TI - A prospective surveillance of drug prescribing and dispensing in a teaching hospital in western Nepal. AB - OBJECTIVE: To evaluate the drug dispensing practices and patients' knowledge on drug use among the outpatients and to identify and analyze the problems in drug prescribing and dispensing. METHODS: A prospective cross-sectional descriptive study was conducted using World Health Organization (WHO) core drug use indicators from July 13, 2008 to August 15, 2008 in Manipal Teaching Hospital, Pokhara, Nepal. RESULTS: A total of 4231 prescriptions were encountered with the total of 10591 drugs prescribed. The average number of drug per prescription was 2.5. Only 13% (n=10591) of drugs were prescribed by generic name. Percentage of drug prescribed from WHO model list of Essential drugs, Essential drug list of Nepal and Nepalese National Formulary was 21.7%, 32.8% and 42.3% respectively. Antibiotics and injections encountered were 28.3% and 3.1% respectively. Average cost per prescription was found to be Nepalese Rupees (NRs) 285.99 (US $ 3.73). Patient knowledge on correct use of drugs and appropriate labeling was found to be 81% and 1.4% respectively. Average dispensing time per prescription was 52 seconds. CONCLUSION: The finding from current study shows a trend towards irrational prescribing and dispensing. Hence, there is a need for effective intervention programme to encourage the physicians and dispensing pharmacists in promoting more rational drug use. PMID- 19813696 TI - Glucagon like peptide-1 receptor agonists and di-peptidyl peptidase inhibitors... A new class of drugs for the treatment of diabetes. PMID- 19813697 TI - Neurosciences: the final frontier. PMID- 19813698 TI - The prevalence of IgA nephropathy in Pakistan: only a tip of the iceberg. PMID- 19813699 TI - Real-time PCR analysis of genes encoding tumor antigens in esophageal tumors and a cancer vaccine. AB - Tumor antigens are the primary target of therapeutic cancer vaccines. We set out to define and compare the expression pattern of tumor antigen genes in esophagus carcinoma biopsies and in an allogeneic tumor lysate-based cancer vaccine, MelCancerVac. Cells used for vaccine production were treated with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-aza-CdR) to determine whether this treatment could improve the profile of tumor antigen genes expressed in these cells. In addition, the presence of MAGE-A tumor antigen protein was evaluated in the purified tumor cell lysate used in the production of the vaccine. Quantitative PCR was used to assay 74 tumor antigen genes in patients with squamous cell carcinoma of the esophagus. 81% (13/16) of tumors expressed more than five cancer/testis (CT) antigens. A total of 96 genes were assayed in the tumor cell clone (DDM1.7) used to make tumor cell lysate for vaccine preparation. Gene expression in DDM1.7 cells was compared with three normal tissues; 16 tumor antigen genes were induced more than ten-fold relative to normal tissues. Treatment with 5-aza-CdR induced expression of an additional 15 tumor antigens to a total of 31. MAGE-A protein was detected in cell lysate by Western blot at an estimated concentration of 0.2 micrograms/ml or 0.01% of the total protein. PMID- 19813700 TI - Thermodynamic perspective on the dock-lock growth mechanism of amyloid fibrils. AB - The mechanism of addition of a soluble unstructured monomer to a preformed ordered amyloid fibril is a complex process. On the basis of the kinetics of monomer disassociation of Abeta(1-40) from the amyloid fibril, it has been suggested that deposition is a multistep process involving a rapid reversible association of the unstructured monomer to the fibril surface (docking) followed by a slower conformational rearrangement leading to the incorporation onto the underlying fibril lattice (locking). By exploiting the vast time scale separation between the dock and lock processes and using molecular dynamics simulation of deposition of the disordered peptide fragment (35)MVGGVV(40) from the Abeta peptide onto the fibril with known crystal structure, we provide a thermodynamic basis for the dock-lock mechanism of fibril growth. Free energy profiles, computed using implicit solvent model and enhanced sampling methods with the distance (delta(C)) between the center of mass of the peptide and the fibril surface as the order parameter, show three distinct basins of attraction. When delta(C) is large, the monomer is compact and unstructured and the favorable interactions with the fibril results in stretching of the peptide at delta(C) approximately 13 A. As delta(C) is further decreased, the peptide docks onto the fibril surface with a structure that is determined by a balance between intrapeptide and peptide fibril interactions. At delta(C) approximately 4 A, a value that is commensurate with the spacing between beta-strands in the fibril, the monomer expands and locks onto the fibril. Using simulations with implicit solvent model and all atom molecular dynamics in explicit water, we show that the locked monomer, which interacts with the underlying fibril, undergoes substantial conformational fluctuations and is not stable. The cosolutes urea and TMAO destabilize the unbound phase and stabilize the docked phase. Interestingly, small crowding particles enhance the stability of the fibril-bound monomer only marginally. We predict that the experimentally measurable critical monomer concentration, C(R), at which the soluble unbound monomer is in equilibrium with the ordered fibril, increases sharply as temperature is increased under all solution conditions. PMID- 19813701 TI - Ordering and freezing-in phenomena of nanochannel water in crystalline organic/inorganic self-assembled complex [Cr(H2bim)3](TMA) x 23.5 H2O. AB - The thermal properties of crystalline complex [Cr(H(2)bim)(3)](TMA) x 23.5 H(2)O were studied by adiabatic calorimetry to clarify the structural ordering and dynamic freezing-in behaviors of the nanochannel water within the pores possessing crystalline wall structure, where H(2)bim denotes 2,2'-biimidazole and TMA is 1,3,5-benzenetricarboxylic acid. Phase and glass transitions were found to occur at 233 K with the associated entropy of Delta(trs)S = 7.96 J K(-1) mol(-1) and at T(g) = 100 K, respectively, in the hydrated sample. The phase transition was interpreted as attributed to the crystallization-like formation of the hydrogen-bond network of the channel-water molecules. The glass transition was interpreted as a freezing-in phenomenon on the way of the development of the network, and its presence indicates that the network formation achieves no completion even at 100 K. The T(g) value is similar to those found previously in other channel-water systems of [Ni(cyclam)(H(2)O)(2)](3)(TMA)(2) x 24 H(2)O and porous silica. It is noted that the channel water within silica pores with their diameter below 1.8 nm undergoes no structural phase transition while the present one does. The origins of the phase and glass transitions and the implication of their presence are discussed based on the difference in the structures of pore wall interacting with the channel-water molecules. PMID- 19813702 TI - Correlations between nucleophilicities and selectivities in the substitutions of tetrahydropyran acetals. AB - Selectivities that deviate from S(N)1 stereoelectronic models in the nucleophilic substitutions of tetrahydropyran acetals were investigated. When weak nucleophiles were employed, stereoselectivities conformed to known S(N)1 stereoelectronic models. In contrast, stereoselectivities in the substitutions of acetals with strong nucleophiles depended on reaction conditions. Erosions in selectivities were observed when strong nucleophiles were employed in the absence of coordinating counterions. These erosions in selectivities are attributed to rates of nucleophilic additions to oxocarbenium ion intermediates that approach the diffusion limit. When triflate counterions were present, however, S(N)2-like pathways became accessible with strong nucleophiles. In most cases examined, the major stereoisomers formed from reactions that proceeded through S(N)2-like pathways were opposite to the major stereoisomers formed from the analogous reactions that proceeded through S(N)1 pathways. PMID- 19813704 TI - Introduction to frontiers in polymer synthesis. PMID- 19813703 TI - De novo design of antimicrobial polymers, foldamers, and small molecules: from discovery to practical applications. AB - Antimicrobial peptides (AMPs) provide protection against a variety of pathogenic bacteria and are, therefore, an important part of the innate immune system. Over the past decade, there has been considerable interest in developing AMPs as intravenously administered antibiotics. However, despite extensive efforts in the pharmaceutical and biotechnology industry, it has proven difficult to achieve this goal. While researchers have solved some relatively simple problems such as susceptibility to proteolysis, more severe problems have included the expense of the materials, toxicity, poor efficacy, and limited tissue distribution. In this Account, we describe our efforts to design and synthesize "foldamers"-- short sequence-specific oligomers based on arylamide and beta-amino acid backbones, which fold into well-defined secondary structures-- that could act as antimicrobial agents. We reasoned that small "foldamers" would be less expensive to produce than peptides, and might have better tissue distribution. It should be easier to fine-tune the structures and activities of these molecules to minimize toxicity. Because the activities of many AMPs depends primarily on their overall physicochemical properties rather than the fine details of their precise amino acid sequences, we have designed and synthesized very small "coarse-grained" molecules, which are far simpler than naturally produced AMPs. The molecular design of these foldamers epitomizes the positively charged amphiphilic structures believed to be responsible for the activity of AMPs. The designed oligomers show greater activity than the parent peptides. They have also provided leads for novel small molecule therapeutics that show excellent potency in animal models for multidrug resistant bacterial infections. In addition, such molecules can serve as relatively simple experimental systems for investigations aimed at understanding the mechanism of action for this class of antimicrobial agents. The foldamers' specificity for bacterial membranes relative to mammalian membranes appears to arise from differences in membrane composition and physical properties between these cell types. Furthermore, because experimental coarse-graining provided such outstanding results, we developed computational coarse-grained models to enable molecular dynamic simulations of these molecules with phospholipid membranes. These simulations allow investigation of larger systems for longer times than conventional molecular dynamics simulations, allowing us to investigate how physiologically relevant surface concentrations of AMP mimics affect the bilayer structure and properties. Finally, we apply the principles discovered through this work to the design of inexpensive antimicrobial polymers and materials. PMID- 19813706 TI - Understanding of the bridging sheet formation of HIV-1 glycoprotein gp120. AB - As the initial step of the entry of HIV-1 into cells, the interaction of CD4 with gp120 is a central area of concern in HIV-1 biology and intervention studies. CD4 binding induces large conformational changes to gp120, such as the formation of the bridging sheet between the V1/V2 stem and beta20/beta21. Understanding the dynamic process and the mechanism that leads to the formation of the bridging sheet is important. We have studied the formation of the bridging sheet via extensive molecular dynamics simulations on a modeled intermediate state. The intermediate state is derived from the crystal structure of the gp120/CD4 complex with rotation of the alpha1 helix and separation of the V1/V2 stem from beta20/beta21. The molecular dynamics simulations reveal that CD4-bound gp120 leads to the refolding of the bridging sheet but the CD4-free gp120 leads to structures similar to unliganded structures of SIV gp120. The bridging sheet refolds with the S375W mutant, but it does not refold with the W112A and S375W/T257S mutants. Our simulation results are in agreement with experimental observations, and they reveal the importance of the formation of the hydrophobic core to the conformational change of CD4-induced gp120. Other experimental observations, including variations in the binding entropy in different mutants, are also qualitatively reproduced. PMID- 19813707 TI - Prediction of the genetic variety of extra virgin olive oils produced at La Comunitat Valenciana, Spain, by fourier transform infrared spectroscopy. AB - Fourier transform infrared spectroscopy (FTIR), followed by multivariate treatment of the spectral data, was used to classify extra virgin olive oils (EVOOs) according to their genetic variety. EVOO samples corresponding to seven different genetic varieties (Arbequina, Borriolenca, Canetera, Farga, Hojiblanca, Picual, and Serrana) were analyzed. The wavelength scale of the FTIR spectra of the oils was divided into 20 regions. The normalized absorbance peak areas within these regions were used as predictor variables. Classification of the EVOO samples according to their genetic variety was achieved by linear discriminant analysis (LDA). A good resolution among all categories was achieved using a LDA model constructed with only nine predictor variables. With this LDA model, 88% of the EVOOs were correctly classified, with assignment probabilities higher than 95%. This method is helpful for olive oil producers because it provides useful information related to the genetic variety of EVOOs, which is required by European Community regulations. PMID- 19813708 TI - Origin of the photo-cross-linking process in dichromated polyacrylamide under conventional and laser irradiation. AB - This work is devoted to determining the contribution of amide groups in the photoredox and cross-linking process of dichromated polyacrylamide based on the fate of the photoactive species and of the polymer under conventional and laser irradiation. It was shown that, in parallel to the reduction of chromium(VI) into chromium(V), the cross-linking of the matrix occurred through a complexation reaction around chromium(V) and through formation of covalent bonds between macromolecular chains. A comparison with dichromated poly(vinyl alcohol) was also reported to highlight the role of the chemical structure of the polymeric matrix in the mechanism of hologram formation. Moreover, for the first time it was demonstrated by in situ infrared spectroscopy that the physicochemical modifications undergone by the photosensitive materials were similar for the two modes of irradiation. PMID- 19813709 TI - Enantioselective synthesis of 1,2,3-trisubstituted cyclopropanes using gem-dizinc reagents. AB - The first asymmetric cyclopropanation of allylic alcohols using gem-dizinc carbenoids, which allows the synthesis of 1,2,3-substituted cyclopropane derivatives in high yields and excellent enantio- and diastereoselectivities, is reported. The initially formed cyclopropylzinc undergoes an in situ B/Zn exchange with the stoichiometric chiral ligand to generate a cyclopropyl borinate that can be directly engaged in a Suzuki-Miyaura cross-coupling reaction. PMID- 19813710 TI - Ab initio study of the structural, tautomeric, pairing, and electronic properties of seleno-derivatives of thymine. AB - The structural, tautomeric, hydrogen-bonding, stacking, and electronic properties of a seleno-derivative of thymine (T), denoted here as 4SeT and created by replacing O4 in T with Se, are investigated by means of ab initio computational techniques. The structural properties of T and 4SeT are very similar, and the geometrical differences are mainly limited to the adjacent environment of the C Se bond. The canonical "keto" form is the most stable tautomer, in the gas phase and in aqueous solution, for both T and 4SeT. It is argued that the competition between two opposite trends, i.e., a decrease in the base-pairing ability and an increase of the stacking interaction upon incorporation of 4SeT into a duplex, likely explains the similar experimental melting points of a seleno-derivative duplex (Se-DNA) and its native counterpart. Interestingly, the underlying electronic structure shows that replacement of O4 with Se promotes a reduction in the HOMO-LUMO gap and an increase in interplane coupling, which suggests that Se DNA could be potentially useful for nanodevice applications. This finding is further supported by the fact that transfer integrals between 4SeT...A stacked base pairs are larger than those determined for similarly stacked natural T...A pairs. PMID- 19813711 TI - The quasicatalytic mechanism: a variation of the catalytic (EC') mechanism. AB - The classic electrochemical catalytic mechanism, often referred to as the EC' mechanism, is traditionally represented by the two reactions A + e <==> B (E(A/B)(0), k(A/B)(0), alpha(A/B)) and B + P <==> A + Q (K(eq), k(f), k(b)). Implicit in this mechanism is the additional heterogeneous electron transfer P + e <==> Q (E(P/Q)(0), k(P/Q)(0), alpha(P/Q)). To observe EC' behavior, the following conditions must be met (we focus on cyclic voltammetric responses): (1) E(P/Q)(0) > E(A/B)(0) (ensuring that K(eq) > 1), (2) k(P/Q)(0)c(P) exp[ alpha(P/Q)(F/RT)(E - E(P/Q)(0))]/(0.446c(A)(FD(A)|v|/RT)(1/2)) << 1 over the potential range of interest (ensuring that the reaction P + e <==> Q does not occur to any significant extent relative to the peak current for reaction A + e <==> B alone), (3) k(f)c(P)RT/F|v| > 1 (ensuring that the catalytic effect is significant). We offer arguments based on Marcus theory that when condition 2 is met, fulfilling condition 3 will be difficult. This could explain why EC' behavior is rare. In the present work we show that EC'-like cyclic voltammetric responses can be obtained even when P + e <==> Q is facile if D(P,Q) (the diffusion coefficient for the substrate-couple species P and Q) is much smaller than D(A,B) (the diffusion coefficient for the mediator-couple species A and B). When D(P,Q)/D(A,B) is sufficiently small, the system behavior becomes identical to that seen for the classical EC' system. We suggest that this "quasicatalytic" behavior should be considered when EC'-like behavior is observed and when the electrochemical system involves a substrate couple whose diffusion coefficients are much smaller than those of the mediator couple. As has been known for some time, when the diffusion coefficients of species A, B, P, and Q are identical (an assumption commonly made to simplify theoretical analysis) and when both heterogeneous electron transfers are reversible, the homogeneous kinetics have no effect on the cyclic voltammetric response--even though the distribution of species in the diffusion layer is dramatically altered. PMID- 19813712 TI - Self-assembly of AB diblock copolymers under confinement into topographically patterned surfaces. AB - Motivated by recent experiments of copolymer patterning by nanoimprinting, we investigate microphase separation and morphology for symmetric AB diblock copolymers with lamellar structure in bulk, confined between a flat bottom surface and a square-wave top surface by using the self-consistent field theory (SCFT). The efficient and high-order accurate pseudospectral method is adopted to numerically solve the SCFT equations in irregularly shaped domains with the help of the "masking" technique by embedding the confined domains of arbitrary shape within a larger rectangular computational cell. Our simulations reveal that the inverted T-style and trapezoid structures occurring in the relatively strong and weak surface fields, respectively, are following our topographically patterned surface. For neutral walls, when the thickness of the lower section is commensurate with the lamellae period of bulk block copolymers, the topographically patterned surface in this work leads to parallel lamellae, and completely parallel lamellae are favored when both the width and height of the upper section are equal to the lamellae bulk period. Furthermore, the prevalent structures are the parallel lamellae in the upper section combined with the perpendicular lamellae in the lower section. When the walls repel one of the block species, parallel lamellae occur in a wide range of film thicknesses compared to the case of neutral walls. To our knowledge, some new structures, however, such as square and partial square structures and reversed-T and trapezoid structures, have not been reported before under parallel surface confinement. In general, the required structures can be obtained by choosing the proper degree of spatial confinement, characterized by variations of the ratio of film thicknesses to bulk repeat period, and the block-substrate interactions. Moreover, we show that the confinement width of the lower section (or the period of the square wave) plays a critical role in microstructure formation. These findings provide a guide to designing novel microstructures involving symmetric diblock copolymers via topographically patterned surfaces and surface fields, relevant to nanoimprinting. PMID- 19813713 TI - California Hass avocado: profiling of carotenoids, tocopherol, fatty acid, and fat content during maturation and from different growing areas. AB - The California Hass avocado ( Persea americana ) is an example of a domesticated berry fruit that matures on the tree during its growing season but ripens only after being harvested. Avocados are typically harvested multiple times during the growing season in California. Previous research has demonstrated potential health benefits of avocados and extracts of avocado against inflammation and cancer cell growth, but seasonal variations in the phytochemical profile of the fruits being studied may affect the results obtained in future research. Therefore, in the present study, avocados were harvested in January, April, July, and September, 2008, from four different growing locations in California (San Luis Obispo, Ventura, Riverside, and San Diego) and analyzed for total fat content, fatty acid profile, carotenoids, and vitamin E. A significant increase in total carotenoid and fat content of avocados from all regions was noted as the season progressed from January to September. Four carotenoids not previously described in the avocado were quantified. The total content of carotenoids was highly correlated with the total fat content (r = 0.99, p < 0.001) demonstrating a remarkable degree of constancy of carotenoid intake per gram of fat content in the California Hass avocado. Future clinical research on the health benefits of the avocado should specify the time of harvest, degree of ripening, growing area, and the total phytochemical profile of the fruit or extract being studied. These steps will enable researchers to account for potential nutrient-nutrient interactions that might affect the research outcomes. PMID- 19813714 TI - Discrimination of small gas molecules through adsorption: reverse selectivity for hydrogen in a flexible metal-organic framework. AB - Zn(2+) ions react with 3,5-pyridinedicarboxylate (pydc) to form a new metal organic framework (MOF), [Zn(pydc)(dma)] (dma = N,N'-dimethylacetamide), based on a noninterpenetrating (10,3)-a topology. The framework possessing narrow one dimensional channels is highly flexible, and as a result, guest-dependent breakthrough-like adsorptions occur under atmospheric pressure. This "gate opening" requires strong interactions between gas molecules and the adsorbent, and therefore [Zn(pydc)(dma)] shows a reverse selectivity for H(2) at 77 K, which is very unusual in MOFs. At 195 K, only CO(2) is selectively adsorbed by this material because of the temperature dependence of the gated adsorption. PMID- 19813715 TI - Epoxyphomalin A and B, prenylated polyketides with potent cytotoxicity from the marine-derived fungus Phoma sp. AB - Chemical investigation of a strain of the marine-derived fungus Phoma sp. has led to the discovery of epoxyphomalin A (1) and B (2), two new prenylated polyketides with unusual structural features. Epoxyphomalin A (1) showed superior cytotoxicity at nanomolar concentrations toward 12 of a panel of 36 human tumor cell lines. In COMPARE analyses, the observed cytotoxic selectivity pattern of 1 did not correlate with those of reference anticancer agents with known mechanisms of action. PMID- 19813716 TI - Evidence for a boroxinate based Bronsted acid derivative of VAPOL as the active catalyst in the catalytic asymmetric aziridination reaction. AB - Studies are described that were designed to determine the structure of the active catalyst in the asymmetric catalytic aziridination of imines with ethyl diazoacetate (AZ reaction). Evidence suggests that the active catalyst contains a boroxine ring in which one of the three boron atoms is spiro-fused with the two phenol groups of the VAPOL ligand. (11)B and (1)H NMR evidence supports the boroxinate structure B in which the counterion to the boroxinate is the protonated form of the imine. The boroxinate structure is also supported by two solid state structures of a VAPOL boroxinate in which the gegen cation is tetramethyl ammonium and 4-dimethylaminopyridinium. PMID- 19813717 TI - Complexation of pinosylvin, an analogue of resveratrol with high antifungal and antimicrobial activity, by different types of cyclodextrins. AB - The complexation of pinosylvin, a potent antimicrobial and antifungal stilbenoid, by cyclodextrins (CDs) is described for first time in this work. Steady-state fluorescence was used to demonstrate that natural (alpha-, beta-, and gamma-CD) and modified (HP-beta-CD, methyl-beta-CD, and ethyl-beta-CD) CDs are able to complex pinosylvin following a 1:1 stoichiometry. However, substantial differences in the strength of the complexation exist between the CDs tested. Although among natural CDs the interaction of pinosylvin with beta-CD was more efficient than with alpha- and gamma-CD, the results show that the complexation constants (K(F)) were higher for all of the modified CDs than for natural CDs, the highest K(F) being that determined for HP-beta-CD-pinosylvin complexes (12112 +/- 761 M(-1)). Moreover, deprotonation of the hydroxyl group of pinosylvin led to a sharp fall in the K(F) values with respect to those observed for the complexes formed between the protonated structure of this stilbenoid and the CDs. Moreover, a pK(a) value is reported for the first time for pinosylvin. Furthermore, when the temperature of the system was increased, a significant drop was observed in the complexation constant values. From these K(F) values and to throw light on the mechanism of pinosylvin affinity for HP-beta-CD, three thermodynamic parameters, DeltaH degrees , DeltaS degrees , and DeltaG degrees , were calculated. The results show that the complexation of pinosylvin by HP-beta CD is a spontaneous and exothermic process with negative values for entropy changes. Finally, to gain information on the effect of the structure of different compounds belonging to the stilbenoid family on the K(F) values, the complexation of other molecules such as (E)-resveratrol and pterostilbene was studied and compared with the results obtained for the HP-beta-CD-pinosylvin complexes. PMID- 19813718 TI - Poly(bisthiophene-carbazole-fullerene) double-cable polymer as new donor-acceptor material: preparation and electrochemical and spectroscopic characterization. AB - A new donor-acceptor dyad, namely, a 3,6-bis(thiophen-2-yl)carbazole derivative bearing a C(60) fullerene as a side group (BTC-F), was prepared and characterized. Electropolymerization of BTC-F leads to the formation of a donor acceptor double-cable polymer (PBTC-F) with high fullerene content (63 wt %) corresponding to one C(60) per polymer repeat unit. The electronic properties of BTC-F and PBTC-F were studied by electrochemical and spectroscopic techniques. Photoluminescence quenching is observed in diluted solutions of BTC-F compared to the nongrafted monomer BTC indicating that an intramolecular charge transfer takes place between the two components of the dyad. The positions of the HOMO and LUMO levels of the monomer and the polymer were accurately determined by differential pulse voltammetry (DPV). The LUMO energy level of the fullerene moiety in BTC-F lies at 3.7 eV below the vacuum level, i.e., slightly higher than corresponding levels of C(60) and PCBM. DPV characterization of PBTC-F indicates little ground state interaction between the pi-conjugated main chain and the C(60) side groups and a high donor HOMO-acceptor LUMO gap of 1.47 eV. PMID- 19813719 TI - The gas phase anisole dimer: a combined high-resolution spectroscopy and computational study of a stacked molecular system. AB - The gas phase structures of anisole dimer in the ground and first singlet electronic excited states have been characterized by a combined experimental and computational study. The dimer, formed in a molecular beam, has been studied by resonance-enhanced multiphoton ionization and high-resolution laser-induced fluorescence techniques. The assignment of the rotational fine structure of the S(1) <-- S(0) electronic transition origin has provided important structural information on the parallel orientation of aromatic rings of anisole moieties. By comparison with the DFT/TD-DFT computational results, it has been possible to infer the detailed equilibrium structure of the complex. The analysis of the equilibrium structure and interaction energy confirms that the anisole dimer is stabilized by dispersive interaction in the gas phase. This is, to the best of our knowledge, the first detailed work (reporting both theoretical and high resolution experimental data) on an isolated cluster in the pi-stacking configuration. PMID- 19813720 TI - Ab initio multiple spawning dynamics of excited butadiene: role of charge transfer. AB - Ab initio multiple spawning simulations of the photochemical reaction dynamics of s-trans-1,3-butadiene were performed. It is found that nonadiabatic events involving two low-lying excited states begin as early as 10 fs after excitation, resulting in the population being split between the bright 1(1)B(u) state and the dark 2(1)A(g) state. The molecule subsequently twists about a terminal carbon carbon bond regardless of whether it is on the 1(1)B(u) or 2(1)A(g) electronic state. This twisting motion leads to conical intersections between S(1) and S(0). Several regions of the intersection seam involving states of differing character are accessed. The regions of the seam involving intersection between a state of charge-transfer character and a state of covalent character dominate the quenching dynamics, but intersections between two covalent states are also accessed a small percentage of the time. The existence and relative energies of these intersections are validated by optimization at the multistate complete active space second-order perturbation level of theory (MS-CASPT2). Our results point to a new mechanism for photoisomerization of butadiene that emphasizes the role of charge-transfer states. PMID- 19813721 TI - Colloidal stability of Al2O3 nanoparticles as affected by coating of structurally different humic acids. AB - The colloidal stability of three structurally different humic acid (HA)-coated Al(2)O(3) nanoparticles (HAs-Al(2)O(3) NPs) was studied in the presence of Ca(2+). HAs were obtained after sequential extractions of Amherst Peat Soil. Highly polar HA1-coated Al(2)O(3) NPs exhibited strong aggregation in the presence of Ca(2+). HA3 and HA7-coated NPs showed weaker aggregation due to their increased aliphaticity and low polarity. HA7-Al(2)O(3) NPs displayed the weakest aggregation behavior even at relatively high Ca(2+) concentration. The inverse stability ratio (alpha = 1/W) was the lowest for HA7-Al(2)O(3) NPs, reflecting that strong steric stabilization enhanced colloidal stability. Atomic force microscopy (AFM) of pure Al(2)O(3) NPs on Ca(2+)-saturated mica clearly demonstrated significant aggregation following classical Derjaguin-Landau-Verwey Overbeek (DLVO) model for hard spheres. On the contrary, weakly polar HA fraction produced approximately 10 nm thick corona of adsorbed layer around each Al(2)O(3) NP, thus stabilizing coated NP suspension through steric effect. Under alkaline conditions and at low ionic strength, adsorbed HA chains swelled, increasing their osmotic potential, which in turn resulted in stabilization of the colloids. Inherent structural variations of natural organic matter (NOM) played a significant part in colloidal stability of the coated NPs. Thus, development of sterically stabilized NPs may have potential application for water remediation in marine and high salinity conditions. PMID- 19813722 TI - Phosphorus-nitrogen compounds. 18. Syntheses, stereogenic properties, structural and electrochemical investigations, biological activities, and DNA interactions of new spirocyclic mono- and bisferrocenylphosphazene derivatives. AB - The reactions of hexachlorocyclotriphosphazatriene, N(3)P(3)Cl(6), with mono- (1 and 2) and bisferrocenyldiamines (3-5), FcCH(2)NH(CH(2))(n)NHR(1) (R(1) = H or FcCH(2)-), produce mono- (6 and 7) and spirocyclic bisferrocenylphosphazenes (8 10). The fully substituted phosphazenes (11-15 and 18-21) are obtained from the reactions of corresponding partly substituted phosphazenes (6-10) with excess pyrrolidine and NH(2)(CH(2))(3)ONa, respectively. The reactions of 6 with 1-aza 12-crown-4 afford geminal (16) and tris (17) crown ether-substituted phosphazenes. The structural investigations of the compounds have been verified by elemental analyses, mass spectrometry, Fourier transform IR, (1)H, (13)C, and (31)P NMR, and DEPT, COSY, HETCOR, and HMBC techniques. The crystal structures of 7, 10, 11, and 15 have been determined by X-ray crystallography. In 16 and 17, there are one and two stereogenic P atoms, respectively, and they are expected to be in enantiomeric mixtures. The structures of 18-21 look similar to a propeller. In 20 and 21, there are two stereogenic P atoms, and they exist as cis (meso; 20a and 21a) and trans (racemic; 20b and 21b) geometric isomers, according to the chiral solvating agent (S)-(+)-2,2,2-trifluoro-1-(9'-anthryl)ethanol experiments. Moreover, the compounds 18 and 19 have three stereogenic P atoms, and they exist as enantiomeric mixtures. Cyclic voltammetric investigations of compounds 6-21a reveal that ferrocene redox centers undergo oxidation concurrently at the same potential with basically reversible peaks, and these compounds appear to be quite robust electrochemically. The compounds 11-15 have been screened for antibacterial activity against gram positive and gram negative bacteria and for antifungal activity against yeast strains.The compounds 11, 12, 14, and 15 are evaluated for antituberculosis activity against reference strain Mycobacterium tuberculosis H37Rv (ATCC 27294). Interactions between compounds 11-15 and pBR322 plasmid DNA are studied by agarose gel electrophoresis. These compounds induce conformational changes in the DNA helix. PMID- 19813723 TI - Systematic thermodynamics of Magneli-phase and other transition metal oxides. AB - Both the molar enthalpies of formation and the absolute entropies of eight transition metal oxides are found to correlate very strongly with their formula unit volumes at room temperature. The metals are Ti, V, Cr, Nb, Mo, Ce, Pr, and Tb. In particular, the thermodynamic values of additive entities (such as TiO(2) in Ti(n)O(2n-1)) in Magneli phases (that is, recombination phases based on rebuilding after shear) are very close to those of the entity as a pure compound. Thus, reliable values of these thermodynamic properties can readily be predicted for unmeasured or even unsynthesized examples, and literature values can be checked. These assertions are checked against published results for which incomplete data is available. The contributions of the disordered regions which form between the added entities is tentatively estimated. PMID- 19813724 TI - Synthesis and gas sorption properties of a metal-azolium framework (MAF) material. AB - A new strut containing an imidazolium tetracarboxylic acid core has been successfully incorporated into a microporous material using paddlewheel coordinated copper(II) ions as nodes. Sorption studies conducted on this permanently microporous material imply that it can separate carbon dioxide from methane with high selectivity. PMID- 19813725 TI - Membrane heterogeneities and fusogenicity in phosphatidylcholine-phosphatidic acid rigid vesicles as a function of pH and lipid chain mismatch. AB - The role of pH-dependent lipid heterogeneities on the fusogenicity of membranes was evaluated on model lipid bilayers in the form of unilamellar vesicles composed of lipid pairs at a fixed equimolar ratio of phosphatidylcholine (PC) and phosphatidic acid (PA) headgroups. The pH and the hydrophobic composition (lipid acyl tails) of membranes were systematically altered, and their effect on vesicle aggregation, membrane fusogenicity, content release, and content mixing was evaluated. Lowering pH increases the extent of protonated PA headgroups forming phase-separated PA-rich heterogeneities and giving rise to molecular packing defects originating at the phase boundaries. Phase boundaries within the membrane's hydrophobic portion are affected by the lipid acyl-tail dynamics (fluidity) and the matching or nonmatching lengths of the acyl tails of lipid pairs comprising the membrane. The aggregates' size increases with lowering pH and is independent of the membrane's hydrophobic composition. Contrary to aggregation, the initial rates of lipid mixing are proportional to pH and also depend on membrane's hydrophobic composition. The apparent lipid-mixing rate constants are greater for membranes containing lipid pairs with mismatched acyl tail lengths, followed by pairs with matching acyl tails in the gel state and by pairs with matching tails where one lipid is close to its transition temperature. Addition of cholesterol conserves the independence of vesicle aggregation from the membrane's hydrophobic composition. However, it decreases the aggregation rates and inverts the tendency for fusion, among the three types of hydrophobic compositions, suggesting a role of cholesterol's preferential partition in different regions of membrane's heterogeneities. We propose a phenomenological model where the membrane phase boundaries containing molecular packing defects act as "sticking points" on the vesicle exterior via which vesicles aggregate upon contact followed by defect merging via intervesicle lipid exchange and mixing. Such heterogeneous bilayers in the form of drug encapsulating liposomes may potentially improve the therapeutic efficacy by fusing with endosomal membranes, thus increasing drug bioavailability. PMID- 19813726 TI - Sequestration, fluorometric detection, and mass spectroscopy analysis of lanthanide ions using surface modified magnetic microspheres for microfluidic manipulation. AB - Several methods for rapid sequestration, fluorometric detection, and the subsequent mass spectroscopic analysis of lanthanide ions using surface modified polystyrene magnetic microspheres are demonstrated. Mixed-ligand antenna complexes of Eu(3+) in which one of the ligands is attached to the surface of the microspheres have been used as a means for the sequestration, immobilization, and detection of these ions. Using the ion-exchange properties of these microspheres, this scheme has been extended to the detection of nonluminescent ions. The principles of these assays form the basis for operation of a portable microfluidic device for general analytical and nuclear forensics applications and indicate the manner in which the established methods of analytical chemistry, such as liquid-liquid extraction and ion-exchange chromatography, can be adapted for such miniature devices. PMID- 19813727 TI - Selective formation of a linear-shaped bundle of microtubules. AB - By using rigid microtubules (MTs) prepared by polymerization with guanylyl (alpha, beta)-methylene-diphosphonate GMPCPP, giant straight-shaped MT bundles were selectively obtained through a dynamic self-assembly process. We demonstrate the effect of the rigidity on the shape and motility of MT bundle composed of GMPCPP-polymerized MTs (GMPCPP-MTs) compared with control MTs that were polymerized with GTP and stabilized with paclitaxel. PMID- 19813728 TI - Identification of absolute helical structures of aromatic multilayered oligo(m phenylurea)s in solution. AB - The oligomeric aromatic ureas bearing N,N'-dimethylated urea bonds such as 3 have aromatic multilayered structure, based on the (cis,cis)-urea structure, and also have dynamic helical structure (all-R or all-S axis chirality) when the benzene rings are connected at the meta positions. The absolute helical structure of oligo(m-phenylurea)s were identified by the empirical and theoretical studies on the CD and vibrational CD (VCD) spectra. Thus, each enantiomer of the oligo(m phenylurea)s 4 bearing a chiral N-2-(methoxyethoxyethoxy)propyl group were synthesized. Intense dispersion-type CD spectra of 4 were observed, which indicated the induction of handedness in the helical structure. In the VCD spectra of 4 in the film state, the signals due to the carbonyl and aromatic ring vibrations were seen with negative and positive values for compounds 4a and 4b, respectively. The calculations of both CD and VCD spectra of oligo(m-phenylurea)s 3 without any chiral N-substituent gave the same assignment about the axis chirality of 4. Thus, the absolute configurations of 4a and 4b are all-R and all S structures, respectively. PMID- 19813729 TI - Expression of Cry3Bb1 in transgenic corn MON88017. AB - To evaluate the effects of transgenic expression of Coleopteran-specific Bt protein Cry3Bb1 on target and nontarget insects in fields with Bt crops, it is necessary to quantify the Cry3Bb1 contents in the plants. Here, we describe the optimization and validation of the quantitative detection of Cry3Bb1 by adapting the commercially available qualitative PathoScreen double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for quantitative measurements. The optimized method had an average accuracy of 84-109% and was used to quantify the Cry3Bb1 contents of different tissues of Bt corn MON88017 at four developmental stages during three years (2005-2007) in a field trial in Germany. The Cry3Bb1 contents were determined based on both dry weight and fresh weight. Cry3Bb1 expression was highest in young leaves (228.4 microg/g dw and 35.5 microg/g fw) and lowest in pollen (3.8 microg/g fw). In root tissues, the Cry3Bb1 content declined during the growing season from 130 to 40 microg/g dw. A significant decline of Cry3Bb1 contents was also observed during the growing season in other plant tissues. The Cry3Bb1 contents of different plant tissues strongly correlated to each other. On the basis of the total corn biomass produced on 1 hectare, it was estimated that up to 905 g of Cry3Bb1 is produced per hectare Bt corn MON88017. PMID- 19813730 TI - Phenylalanine ammonia-lyase, polyphenol oxidase, and phenol concentration in fruits of Olea europaea L. cv. Picual, Verdial, Arbequina, and Frantoio during ripening. AB - The kinetics and protein-expression level of phenylalanine ammonia-lyase (PAL) and polyphenol oxidase (PPO) in fruits of olive trees (Olea europaea) cv. Picual, Verdial, Arbequina, and Frantoio have been studied in relation to the concentration of total phenolic compounds, oleuropein, hydroxytyrosol, and tyrosol during fruit ripening. Frantoio was the variety that showed the highest total phenol concentration, the highest PAL activity, the lowest PPO activity, and the lowest protein levels. In contrast, Verdial was the variety that showed the lowest total phenol concentration, the least PAL activity, the greatest PPO activity, and the highest protein levels. Arbequina and Picual showed intermediate levels. These results suggest the existence of a coordinated response between PAL, PPO, and the concentration of total phenols over ripening in the four varieties. The concentration of total and specific phenols differed between varieties and specifically changed over ripening. PMID- 19813731 TI - Study on the effects of heating of virgin olive oil blended with mildly deodorized olive oil: focus on the hydrolytic and oxidative state. AB - In this study, mildly deodorized olive oil (DEO) and its admixtures with extra virgin olive oil (EVOO) have been analyzed after heating treatments by microwave and conventional oven. Different patterns in oxidative and hydrolytic degradation of lipids in genuine and sophisticated olive oils have been evaluated by chromatographic and spectroscopic methods and related to heating treatments. The experimental plan focused on the assessment of the (a) hydrolytic degree of the samples by the free acidity and the 1,2- and 1,3-diacylglycerols (DAG) determinations; (b) oxidative status of the samples by the assessment of the peroxide value (POV) and oxidized fatty acid (OFA), the specific absorption at 270 nm (k(270)), the accelerated aging test (OSI) and volatile compounds. In general, the thermal treatment by conventional oven led to a higher content of 1,3-DAG and secondary oxidation products than microwave heating. A duo-trio sensory test was also performed: tasters were not able to discriminate between EVOO and DEO heated by conventional oven whereas they were when oils were microwaved. PMID- 19813732 TI - Blocking peptides decrease tissue transglutaminase processing of gliadin in vitro. AB - Tissue transglutaminase (tTG) plays an important role in celiac disease pathology as it catalyzes deamidation and cross-linking of specific gluten peptides and converts them into potent epitopes recognized by intestinal T-cells. We investigated whether synthetic peptides with high affinity to gliadin could alter tTG activity on gliadin and whole gluten digest. The immobilized substrates were incubated with synthetic peptides identified by the phage display technique and a control peptide with no affinity to gliadin. Transglutaminase activity was measured with time resolved fluorescence. The mean tTG activity, compared to that of the control without the peptides, was reduced by 31, 33, and 36% for three selected gliadin-binding peptides, and 30% for the peptide pool (P < or = 0.001 0.004) when gliadin was the substrate. Finally, substrate specificity experiments suggested that avenin was processed in a manner similar that used for gliadin during in vitro assays with tTG. The results showed that the blocking peptides efficiently reduced tTG processing of gliadin in vitro, and this strategy will be further investigated as an alternative therapy for celiac disease. PMID- 19813733 TI - Effect of cosolvents on the structural stability of endoglucanase from Aspergillus aculeatus. AB - The effects of cosolvents such as sucrose, glycerol, and sorbitol on endoglucanase have been studied by activity, circular dichroic spectroscopy, fluorescence, and apparent thermal transition temperature measurements. The endoglucanase activity increased by 4-fold at 40% cosolvent concentration under optimum conditions. The endoglucanase lost 50% of its activity when exposed to 90 degrees C for more than 30 min (1 h). In the presence of cosolvents, it maintained its original activity and native conformation as indicated by far UV CD at 70 degrees C. The app T(m) increased from a control value of 57 degrees C to a value of 66 degrees C in the presence of 40% sucrose. The partial specific volume of endoglucanase was 0.723 mL/g in sodium acetate buffer. The preferential interaction parameters were negative in all cosolvents, and the maximum hydration of the protein was observed in 40% glycerol where the preferential interaction parameter was -0.126 g/g. The thermal stability of endoglucanase increased in the presence of cosolvents. PMID- 19813734 TI - Effects of ionic liquids on the characteristics of synthesized nano Fe(0) particles. AB - In this paper, high specific surface area zerovalent Fe(0) nanoparticles with unusual morphology and high reductive activity were prepared using a chemical reductive reaction of iron chloride hydrate with sodium borohydride in an aqueous solution of ionic liquids (ILs). The ionic liquids, [C(4)mim]X (X = Cl(-), Br(-), BF(4)(-), PF(6)(-)) and [C(n)mim][BF(4)] (n = 4, 6, 8), were used for this purpose. Transmission electron microscopy, X-ray diffraction, and Brunauer-Emmett Teller surface area measurements were used to characterize the morphology, structure, and physical properties of the as-prepared Fe(0). The results suggest that the nucleation and growth of the Fe(0) were governed by an IL-controlled reductive reaction mechanism. The effects of ionic liquids (cations, anions, and concentration) on the morphology of the Fe(0) were examined. It was shown that anions of the ILs played a key role in the morphology and size of Fe(0), and the nanoparticle size of the Fe(0) prepared in the presence of different ionic liquids follows the trend: [C(4)mim]Cl > [C(4)mim]Br > [C(4)mim][BF(4)] > [C(4)mim][PF(6)] and [C(4)mim][BF(4)] > [C(6)mim][BF(4)] > [C(8)mim][BF(4)]. The reductive activity of the iron nanoparticles was studied by the denitrification experiment of sodium nitrite. The removal rate of nitrite by the IL-capped Fe(0) was found to be much faster than that by the noncapped Fe(0). This indicates that the IL-capped Fe(0) particles are better candidates for the reductive degradation. The high reductive activity of the Fe(0) nanoparticles was attributed to their high surface area and more active sites. PMID- 19813735 TI - Charge mobility and transport behavior in the ordered and disordered states of the regioregular poly(3-hexylthiophene). AB - We theoretically analyze the charge-transfer behavior of regioregular poly(3 hexylthiophene) (rr-P3HT) by quantum mechanical (QM) and molecular dynamics (MD) methods. In particular, we clarify the effects associated with the respective contribution from the ordered and disordered regions. In the ordered regions, the typical value of the hole mobility along the intrachain route is about 1 cm(2) V( 1) s(-1), which is significantly larger than that along the pi-pi interchain route, approximately 10(-2) cm(2) V(-1) s(-1). Our results indicate that the main charge-transfer route within the P3HT ordered lamellae is along the intrachain direction instead of the interchain direction. Moreover, the calculated hole mobility of 10(-2) cm(2) V(-1) s(-1) along the pi-pi interchain route is consistent with the experimental data measured in the P3HT single fibril. In the disordered regions, we propose a crossing-point/bridging-chain model to describe the charge-transport routes. In this model, the hole mobility can reach the limit of around 10(-2) and 1 cm(2) V(-1) s(-1) when the charge takes the interchain route through the crossing points and the intrachain route along the bridging chains, respectively. As expected, the resultant mobility in the disordered state is strongly affected by the ratio of the amount of crossing points and bridging chains. When considering the presence of both ordered and disordered regions, the average overall charge mobility is mainly dominated by the charge transport in the disordered regions. The fact that some of the experimentally measured hole mobility by varying the molecular weight is limited to a maximum value of 10(-2) cm(2) V(-1) s(-1) is mainly due to the presence of more crossing points instead of the bridging chains in the disordered regions. With increasing the amount of the bridging chains in the disordered regions, one can expect an enhancement of the charge mobility, such as to the experimentally obtained high value of 0.1 cm(2) V(-1) s(-1). PMID- 19813736 TI - A versatile new synthetic route to 1N-hydroxyindazoles. AB - A new and versatile cyclization reaction affording rare 1N-hydroxyindazoles is presented. Treatment of 2-nitrobenzylamines with methanolic sodium hydroxide furnishes 1N-hydroxyindazoles regioselectively and in high yield. The reaction tolerates a range of functional groups and electronic effects. PMID- 19813737 TI - Synthesis and photophysics of ambipolar fluoren-9-ylidene malononitrile derivatives. AB - The efficient synthesis of ambipolar 2,7- and 3,6-disubstituted fluoren-9-ylidene malononitrile derivatives (4 and 9) is described. Structure-activity relationships depend on the position of substitution. Population of the S(1) excited state in the 2,7-disubstituted FM derivatives is achieved via higher states, as evidenced from the UV-vis absorption and emission spectra. The results are supported by TDDFT calculations. Charge transfer states were the main deactivation path of the excited states of ambipolar 9b in polar solvents, as evidenced by fluorescence spectroscopy. PMID- 19813738 TI - Oxidation of cyclohexane by high-valent iron bispidine complexes: tetradentate versus pentadentate ligands. AB - The iron-bispidine-catalyzed oxidation of cyclohexane with H(2)O(2), where either a tetradentate or a pentadentate bispidine ligand is coordinated to the iron center, yields up to 35% cyclohexanol and cyclohexanone (alcohol/ketone ratio of up to 4). Product distribution (including (18)O labeling studies), kinetic isotope effects, and the ratio of tertiary/secondary alcohols with adamantane as a substrate (tertiary/secondary) suggest that (i) H abstraction by a ferryl complex is the rate-determining step and that the emerging cyclohexyl radical is short-lived, (ii) there is a parallel reaction involving oxidation by OH radicals, and (iii) there are considerable differences in the reaction pathways between the tetradentate and pentadentate ligand catalyst. These interpretations are fully supported by a DFT-based computational analysis. PMID- 19813739 TI - Effects of urea and trimethylamine-N-oxide on the properties of water and the secondary structure of hen egg white lysozyme. AB - The influence of urea and trimethylamine-N-oxide (TMAO) on the structure of water and secondary structure of hen egg white lysozyme (HEWL) has been investigated. The hydration of these osmolytes was studied in aqueous solutions by means of FTIR spectra of HDO isotopically diluted in H(2)O. The difference spectra procedure was applied to remove the contribution of bulk water and thus to separate the spectra of solute-affected HDO. The structural-energetic characteristic of these solute-affected water molecules shows that, on average, water affected by TMAO forms stronger H-bonds and is more ordered than pure water. In the case of urea, the H-bonds are very similar to those in pure water. To facilitate the interpretation of the obtained spectral results, calorimetric measurements, DFT calculations, and molecular dynamics (MD) simulations of aqueous osmolyte clusters were performed. All of these results confirmed that the interactions of TMAO with water molecules are much stronger than those of urea with water. Additional ATR FTIR measurements were performed to characterize the influence of the examined osmolytes on the secondary structure of HEW lysozyme. The type of interactions (direct or indirect) was determined, based on the second derivatives of ATR protein spectra record during an increase in the osmolyte concentration. The changes in the amide I band shape caused by urea or TMAO were found to correlate quite well with changes in the water structure around these osmolytes. PMID- 19813740 TI - Photoionization of three isomers of the C9H7 radical. AB - Three resonance-stabilized radicals, 1-indenyl (Ind), 1-phenylpropargyl (1PPR), and 3-phenylpropargyl (3PPR), all isomers of the composition C(9)H(7), were generated by jet flash pyrolysis. Their photoionization was examined by VUV synchrotron radiation. The mass spectra show a clean and efficient radical generation when the pyrolysis is turned on. To study the photoionization, photoion yield measurements and threshold photoionization spectroscopy techniques were applied. We determined adiabatic ionization energies (IE(ad)) of 7.53 eV for Ind, 7.20 eV for 3PPR, and 7.4 eV for 1PPR. Ab initio calculations show no major change in geometry upon ionization, in agreement with ionization from a nonbonding molecular orbital. The IEs were also computed and are in agreement with the measured ones. The difference in the IE might allow a distinction of the three isomers in flames. In the indenyl spectrum, an excited a(+) (3)B(2) state of the cation was identified at 8.10 eV, which shows a low-energy vibrational progression of 61 meV. Furthermore, we have examined the dissociative photoionization of the precursors. The indenyl precursor, 1-indenyl bromide, undergoes dissociative photoionization to Ind(+). An appearance energy (AE(0K)) of 10.2 eV was obtained from fitting the experimental breakdown diagram. A binding energy of 1.8 eV can thus be determined for the C-Br bond in 1-indenyl bromide. The phenylpropargyl precursors 1PPBr (1-phenylpropargyl bromide/3-phenyl 3-bromopropyne) and 3PPBr (3-phenylpropargyl bromide/1-phenyl-3-bromopropyne) also lose a bromine atom upon dissociative photoionization. Approximate appearance energies of 9.8 eV for 3PPBr and 9.3 eV for 1PPBr have been determined. PMID- 19813741 TI - Application of Negishi cross-coupling to the synthesis of the cyclic tripeptides OF4949-III and K-13. AB - Syntheses of the cyclic tripeptides OF4949-III 1 and K-13 2 are reported, in which the key steps are intermolecular and intramolecular Negishi cross-coupling reactions, respectively. In addition, the synthesis of a protected isomer of K-13 25 is reported. The synthesis of K-13 features a tripeptidic organozinc reagent 11, one of the most highly functionalized such reagents to be described. An O aryltyrosine derivative 15, prepared by S(N)Ar reaction between Boc-tyrosine and 2-fluorobenzaldehyde, followed by Dakin reaction, iodination, and methylation, is used as a common intermediate for all of the syntheses described. The routes to this class of cyclic tripeptide are among the shortest reported to date and demonstrate the high functional group tolerance of the carbon-zinc bond toward peptide derivatives. PMID- 19813742 TI - Single molecule spectroscopy reveals heterogeneous transport mechanisms for molecular ions in a polyelectrolyte polymer brush. AB - Single molecule polarization and fluorescence correlation spectroscopy were used to evaluate heterogeneous transport mechanisms of molecular ions within supported polyelectrolyte brushes. Modes of diffusive transport include periods of significantly restricted rotational motion, often maintained over tens of milliseconds; periods of fast molecular rotation; and occasional adsorption of fluorescent probe molecules in the brush. The studies reveal rapid switching between orientational states during each observed mode of motion. Through quantitative analysis of state occupation times, the rate constants for transitions from weakly associated to strongly associated states were extracted. Additionally, the pH dependence of the ion transport rates in the brush exhibits an abrupt, rather than continuous, trend. These single molecule studies demonstrate the presence of dynamic anisotropic interactions between the charged molecular probe and the polymer brush and provide experimental evidence of stimuli responsive switchable transport functionality in the polyelectrolyte brush. PMID- 19813743 TI - Biphenyls from Berberis koreana. AB - Three new biphenyls, berbekorin A (1), 2'-hydroxy-3,4,5-trimethoxybiphenyl (2), and 4,5-dihydroxy-3-methoxybiphenyl (3), together with 11 known biphenyls (4-14), were isolated from the n-hexane-soluble fraction of the MeOH extract of the trunk of Berberis koreana. The structures of 1-3 were determined by spectroscopic methods, including 1D and 2D NMR analysis. Compound 1 exhibited cytotoxic activity against the SK-MEL-2 skin melanoma cell line, in particular. Moreover, compounds 2, 5, 6, and 14 inhibited NO production in LPS-activated BV-2 cells, a microglial cell line. PMID- 19813744 TI - Probing compositional variation within hybrid nanostructures. AB - We present a detailed analysis of the structural and magnetic properties of solution-grown PtCo-CdS hybrid structures in comparison to similar free-standing PtCo alloy nanoparticles. X-ray absorption spectroscopy is utilized as a sensitive probe for identifying subtle differences in the structure of the hybrid materials. We found that the growth of bimetallic tips on a CdS nanorod substrate leads to a more complex nanoparticle structure composed of a PtCo alloy core and thin CoO shell. The core-shell architecture is an unexpected consequence of the different nanoparticle growth mechanism on the nanorod tip, as compared to free growth in solution. Magnetic measurements indicate that the PtCo-CdS hybrid structures are superparamagnetic despite the presence of a CoO shell. The use of X-ray spectroscopic techniques to detect minute differences in atomic structure and bonding in complex nanosystems makes it possible to better understand and predict catalytic or magnetic properties for nanoscale bimetallic hybrid materials. PMID- 19813745 TI - Structure and chemical composition of layers adsorbed at interfaces with champagne. AB - The structure and the chemical composition of the layer adsorbed at interfaces involving champagne have been investigated using native champagne, as well as ultrafiltrate (UFch) and ultraconcentrate (UCch) obtained by ultrafiltration with a 10(4) nominal molar mass cutoff. The layer adsorbed at the air/liquid interface was examined by surface tension and ellipsometry kinetic measurements. Brewster angle microscopy demonstrated that the layer formed on polystyrene by adsorption or drop evaporation was heterogeneous, with a domain structure presenting similarities with the layer adsorbed at the air/liquid interface. The surface chemical composition of polystyrene with the adlayer was determined by X-ray photoelectron spectroscopy (XPS). The contribution of champagne constituents varied according to the liquid (native, UFch, and UCch) and to the procedure of adlayer formation (evaporation, adsorption, and adsorption + rinsing). However, their chemical composition was not significantly influenced either by ultrafiltration or by the procedure of deposition on polystyrene. Modeling this composition in terms of classes of model compounds gave approximately 35% (w/w) of proteins and 65% (w/w) of polysaccharides. In the adlayer, the carboxyl groups or esters represent about 18% of carbon due to nonpolypeptidic compounds, indicating the presence of either uronic acids in the complex structure of pectic polysaccharides or of polyphenolic esters. This structural and chemical information and its relationship with the experimental procedures indicate that proteins alone cannot be used as a realistic model for the macromolecules forming the adsorption layer of champagne. Polysaccharides, the other major macromolecular components of champagne wine, are assembled with proteins at the interfaces, in agreement with the heterogeneous character of the adsorbed layer at interfaces. PMID- 19813746 TI - Phthalides by rhodium-catalyzed ketone hydroacylation. AB - Phthalides are biologically relevant five-membered lactones found in herbs, fruits, and vegetables. Herein we communicate the first atom-economical approach to phthalides by using enantioselective ketone hydroacylation. In the presence of Rh[(Duanphos)]X (X = NO(3), OTf, OMs), various 2-ketobenzaldehydes undergo intramolecular hydroacylation to produce phthalide products in good yields and 92 98% ee's. Our study highlights the key role counterions play in controlling both reactivity and enantioselectivity. A concise asymmetric total synthesis of the celery extract (S)-(-)-3-n-butylphthalide is also presented. PMID- 19813747 TI - Toward a rational design of multitarget-directed antioxidants: merging memoquin and lipoic acid molecular frameworks. AB - Novel multitargeted antioxidants 3-6 were designed by combining the antioxidant features, namely, a benzoquinone fragment and a lipoyl function, of two multifunctional lead candidates. They were then evaluated to determine their profile against Alzheimer's disease. They showed antioxidant activity, improved following enzymatic reduction, in mitochondria and T67 cell line. They also displayed a balanced inhibitory profile against amyloid-beta aggregation and acetylcholinesterase, emerging as promising molecules for neuroprotectant lead discovery. PMID- 19813748 TI - 405 nm absorption detection in nanoliter volumes. AB - Analytical UV absorption detection for microfluidic devices, capillary electrophoresis, and even high-performance liquid chromatography is hampered by the small detection volumes, short absorption paths, and the need to sample at a high rate with a stable background and low noise. Fiber-loop ring-down spectroscopy (FLRDS) permits absorption detection of dilute liquid samples in volumes as small as a few nanoliters, while being insensitive to light source fluctuations and permitting a millisecond temporal resolution. We demonstrate a FLRDS based detection scheme that is compatible in dimensions (<200 microm absorption path, 6.0 nL detection volume) and optical design (405 nm detection wavelength, fiber coupled) with existing separation systems. An optical/fluidic interface has been built that allows injection of laser light into the loop while also permitting delivery of the sample. The detection limit of tartrazine was determined to be 5 microM (30 fmol) corresponding to an absorption of 0.11 cm( 1). Equivalent results were obtained when detecting myoglobin, a heterocyclic pharmaceutical ingredient, and 5.17 microm diameter polystyrene beads. PMID- 19813749 TI - P450 enzymes: their structure, reactivity, and selectivity-modeled by QM/MM calculations. PMID- 19813750 TI - Highly regioselective N-2 arylation of 4,5-dibromo-1,2,3-triazole: efficient synthesis of 2-aryltriazoles. AB - Reaction of 4,5-dibromo-1,2,3-triazole with electron-deficient aromatic halides in the presence of potassium carbonate in DMF produces the corresponding 2-aryl 4,5-dibromotriazoles with high regioselectivity. Subsequent debromination of these triazoles by hydrogenation furnishes 2-aryltriazoles in excellent yields. Overall, this two-step process provides an efficient access to 2-aryl-1,2,3 triazoles. PMID- 19813751 TI - An asymmetric ortholithiation approach to inherently chiral calix[4]arenes. AB - A general asymmetric synthesis of inherently chiral calix[4]arenes is described: using a chiral oxazoline derived from L-valine, an ortholithiation strategy is employed to give inherently chiral calix[4]arenes with high (93%) enantiomeric excesses. A crystal structure of a phosphine oxide intermediate has been obtained, unambiguously assigning the major diastereomer in the reaction; a mechanism explaining this result is proposed. PMID- 19813752 TI - Highly enantioselective catalytic 1,3-dipolar cycloaddition involving 2,3 allenoate dipolarophiles. AB - A bisphosphoric acid-catalyzed 1,3-dipolar cycloaddition of buta-2,3-dienoates with azomethine ylides yields 3-methylenepyrrolidine derivatives with excellent enantioselectivity (up to 97% ee). PMID- 19813753 TI - Highly efficient and tunable synthesis of dioxabicyclo[4.2.1] ketals and tetrahydropyrans via gold-catalyzed cycloisomerization of 2-alkynyl-1,5-diols. AB - A highly efficient gold(I) chloride catalyzed cycloisomerization of 2-alkynyl-1,5 diol (1) to dioxabicyclo[4.2.1] ketal (2) and its further transformation to tetrahydropyran (3) are reported. The diol is readily obtained by the reduction of 2-alkynyl-substituted glutarates, isolated from the Michael addition of allenoates to methyl acrylate. These reactions proceeded smoothly under very mild conditions. A plausible mechanism for the formation of the said tetrahydropyran from the corresponding ketal is proposed. PMID- 19813754 TI - Inhibition of the dimerization and active site of HIV-1 protease by secondary metabolites from the Vietnamese mushroom Ganoderma colossum. AB - A new farnesyl hydroquinone, ganomycin I (1), was isolated along with ganomycin B (2) from the chloroform extract of the fruiting bodies of the Vietnamese mushroom Ganoderma colossum. These compounds inhibited HIV-1 protease with IC50 values of 7.5 and 1.0 microg/mL, respectively. Kinetic studies using Zhang-Poorman and Lineweaver plots revealed that compound 2 competitively inhibited the active site of the enzyme, whereas the tetracyclic triterpene schisanlactone A, previously isolated from the same fungus, was a dimerization inhibitor, with an IC50 value of 5.0 microg/mL. The previous findings were also confirmed by the virtual docking of both compounds with HIV-1 protease crystal structure. PMID- 19813755 TI - Ultrasmall mode volume plasmonic nanodisk resonators. AB - We study the resonant modes of nanoscale disk resonators sustaining metal insulator-metal (MIM) plasmons and demonstrate the versatility of these cavities to achieve ultrasmall cavity mode volume. Ag/SiO2/Ag MIM structures were made by thin-film deposition and focused ion beam milling with cavity diameters that ranged from d = 65-2000 nm. High-resolution two-dimensional cavity-mode field distributions were determined using cathodoluminescence imaging spectroscopy and are in good agreement with boundary element calculations. For the smallest cavities (d = 65-140 nm), the lowest order mode (m = 1, n = 1) is observed in the visible spectral range. This mode is of similar nature as the one in plasmonic particle dimers, establishing a natural connection between localized and traveling plasmon cavities. A cavity quality factor of Q = 16 is observed for the 105 nm diameter cavity, accompanied by a mode volume as small as 0.00033lamda(0)(3). The corresponding Purcell factor is 900, making these ultrasmall disk resonators ideal candidates for studies of enhanced spontaneous emission and lasing. PMID- 19813756 TI - Light-controlled one-sided growth of large plasmonic gold domains on quantum rods observed on the single particle level. AB - We create large gold domains (up to 15 nm) exclusively on one side of CdS or CdSe/CdS quantum rods by photoreduction of gold ions under anaerobic conditions. Electrons generated in the semiconductor by UV stimulation migrate to one tip where they reduce gold ions. Large gold domains eventually form; these support efficient plasmon oscillations with a light scattering cross section large enough to visualize single hybrid particles in a dark-field microscope during growth in real time. PMID- 19813757 TI - Theoretical characterization of an atmospheric pressure glow discharge used for analytical spectrometry. AB - We have investigated the plasma processes in an atmospheric pressure glow discharge (APGD) in He used for analytical spectrometry by means of fluid and Monte Carlo (MC) simulations. Typical results include the potential and electric field distributions in the plasma, the density profiles of the various plasma species throughout the discharge, the mean electron energy, as well as the rates of the various collision processes in the plasma, and the relative importance of the different production and loss rates for the various species. The similarities and differences with low-pressure glow discharges are discussed. The main differences are a very small cathode dark space region and a large positive column as well as the dominant role of molecular ions. Some characteristic features of the APGD, such as the occurrence of the different spatial zones in the discharge, are illustrated, with links to experimental observations. PMID- 19813758 TI - ent-Pimarane diterpenoids from Siegesbeckia orientalis and structure revision of a related compound. AB - Six new ent-pimarane diterpenoids, namely, ent-14beta,16-epoxy-8-pimarene 3beta,15alpha-diol (1), 7beta-hydroxydarutigenol (2), 9beta-hydroxydarutigenol (3), 16-O-acetyldarutigenol (4), 15,16-di-O-acetyldarutoside (5), and 16-O acetyldarutoside (6), were isolated from the ethanol extract of Siegesbeckia orientalis. Their structures were elucidated on the basis of spectroscopic studies, and the absolute configuration of 1 was established by the modified Mosher's method. Furthermore, the structure of ent-12beta,16-epoxy-8-pimarene 2alpha,15beta,19-triol, recently isolated from the same plant, should be revised as ent-14beta,16-epoxy-8-pimarene-2alpha,15alpha,19-triol (10). PMID- 19813759 TI - Mechanism of formation of (deoxy)guanosine adducts derived from peroxidase catalyzed oxidation of the carcinogenic nonaminoazo dye 1-phenylazo-2 hydroxynaphthalene (Sudan I). AB - We investigated peroxidase-mediated oxidation of and the formation of the (deoxy)guanosine adduct by 1-phenylazo-2-hydroxynaphthalene (Solvent Yellow 14, Sudan I), a liver and urinary bladder carcinogen for rodents and a potent contact allergen and sensitizer for humans. Using thin layer chromatography (TLC) and/or high performance liquid chromatography (HPLC) combined with mass and/or nuclear magnetic resonance (NMR) spectrometry, we characterized the structures of two major peroxidase-mediated Sudan I metabolites and those of the adducts of (deoxy)guanosine that are formed during Sudan I oxidation. Peroxidase oxidizes Sudan I to radical species that react with another Sudan I radical to form the Sudan I dimer, or in the presence of (deoxy)guanosine, the oxidized Sudan I can attack the exocyclic amino group of guanine, forming the 4-[(deoxy)guanosin-N(2) yl]Sudan I adduct. The reaction product with a second Sudan I radical results in a dimer where the oxygen 2 radical of Sudan I reacted with carbon 1 in the second Sudan I skeleton. The Sudan I dimer is unstable and decomposes spontaneously to the second oxidation product. This compound consists of the 4-oxo-Sudan I skeleton connected via the oxygen of its 2-hydroxyl group and nitrogen of its azo group with carbon 1 of 2-oxonaphthalene, having a unique spironaphthooxadiazine structure. If (deoxy)guanosine is present during the formation of this Sudan I metabolite, an adduct, in which this Sudan I metabolite is bound to the exocyclic amino group of guanine, is generated. This (deoxy)guanosine adduct is again unstable and decomposes spontaneously to the same adduct that is formed by the direct reaction of oxidized Sudan I, the 4-[(deoxy)guanosin-N(2)-yl]Sudan I adduct. The results presented here are the first structural characterization of Sudan I-(deoxy)guanosine adducts formed during the oxidation of this carcinogen by peroxidase. PMID- 19813760 TI - Requirement of a unique Ca(2+)-binding loop for folding of Tk-subtilisin from a hyperthermophilic archaeon. AB - Tk-subtilisin from the hyperthermophiolic archaeon Thermococcus kodakaraensis matures from Pro-Tk-subtilisin upon autoprocessing and degradation of Tk propeptide [Tanaka, S., Saito, K., Chon, H., Matsumura, H., Koga, Y., Takano, K., and Kanaya, S. (2007) J. Biol. Chem. 282, 8246-8255]. It requires Ca(2+) for folding and assumes a molten globule-like structure in the absence of Ca(2+) even in the presence of Tk-propeptide. Tk-subtilisin contains seven Ca(2+)-binding sites. Four of them (Ca2-Ca5) are located within a long loop, which mostly consists of a unique insertion sequence of this protein. To analyze the role of this Ca(2+)-binding loop, three mutant proteins, Deltaloop-Tk-subtilisin, DeltaCa2-Pro-S324A, and DeltaCa3-Pro-S324A, were constructed. These proteins were designed to remove the Ca(2+)-binding loop, Ca2 site, or Ca3 site of Pro-Tk subtilisin or its active site mutant Pro-S324A. Far-UV CD spectra of these proteins refolded in the absence and presence of Ca(2+) indicated that Deltaloop Tk-subtilisin completely lost the ability to fold into a native structure. In contrast, two other proteins retained this ability, although their refolding rates were greatly decreased compared to that of Pro-S324A. Determination of the crystal structures of these proteins purified in a Ca(2+)-bound form indicates that the structures of DeltaCa2-Pro-S324A and DeltaCa3-Pro-S324A are virtually identical to that of Pro-S324A, except that they lack the Ca2 and Ca3 sites, respectively, and the structure of the Ca(2+)-binding loop is destabilized. Nevertheless, these proteins were slightly more stable than Pro-S324A. These results suggest that the Ca(2+)-binding loop is required for folding of Tk subtilisin but does not seriously contribute to the stabilization of Tk subtilisin in a native structure. PMID- 19813762 TI - Classification of cytochrome p(450) activities using machine learning methods. AB - The cytochrome P(450) (CYP) system plays an integral part in the metabolism of drugs and other xenobiotics. Knowledge of the structural features required for interaction with any of the different isoforms of the CYP system is therefore immensely valuable in early drug discovery. In this paper, we focus on three major isoforms (CYP 1A2, CYP 2D6, and CYP 3A4) and present a data set of 335 structurally diverse drug compounds classified for their interaction (as substrate, inhibitor, or any interaction) with these isoforms. We also present machine learning models using a variety of commonly used methods (k-nearest neighbors, decision tree induction using the CHAID and CRT algorithms, random forests, artificial neural networks, and support vector machines using the radial basis function (RBF) and homogeneous polynomials as kernel functions). We discuss the physicochemical features relevant for each end point and compare it to similar studies. Many of these models perform exceptionally well, even with 10 fold cross-validation, yielding corrected classification rates of 81.7 to 91.9% for CYP 1A2, 89.2 to 92.9% for CYP 2D6, and 87.4 to 89.9% for CYP3A4. Our models help in understanding the structural requirements for CYP interactions and can serve as sensitive tools in virtual screenings and lead optimization for toxicological profiles in drug discovery. PMID- 19813761 TI - Structure of chemokine-derived antimicrobial Peptide interleukin-8alpha and interaction with detergent micelles and oriented lipid bilayers. AB - Interleukin-8alpha (IL-8alpha) is an antimicrobial peptide derived from the chemokine IL-8. Solution NMR was used to determine the atomic-resolution structure of IL-8alpha in SDS micelles. Solid-state NMR and tryptophan fluorescence were used to probe the interaction of IL-8alpha with model membranes. The peptide interacted differently with anionic versus purely zwitterionic micelles or bilayers. Tryptophan fluorescence demonstrated a deeper position of Trp4 in SDS micelles and POPC/POPG bilayers compared to pure POPC bilayers, consistent with (2)H order parameters, which also indicated a deeper position of the peptide in POPC/POPG bilayers compared to POPC bilayers. Paramagnetic probe data showed that IL-8alpha was situated roughly parallel to the SDS micelle surface, with a slight tilt that positioned the N-terminus more deeply in the micelle compared to the C-terminus. (15)N solid-state NMR spectra indicated a similar, nearly parallel position for the peptide in POPC/POPG bilayers. (31)P and (2)H solid-state NMR demonstrated that the peptide did not induce the formation of any nonlamellar phases and did not significantly disrupt bilayer orientation in aligned model membranes composed of POPC or POPC and POPG. PMID- 19813763 TI - Estradiol and progesterone-mediated regulation of P-gp in P-gp overexpressing cells (NCI-ADR-RES) and placental cells (JAR). AB - The effect of progesterone and estrogen treatment on the expression and function of P-glycoprotein (P-gp) was evaluated in JAR cells and a P-gp overexpressing cell line, NCI-ADR-RES. Western blot analysis and real-time Q-PCR were used to evaluate P-gp protein and MDR1 mRNA expression respectively in the cells following incubation with progesterone (P4) and/or beta-estradiol (E2). Cellular uptake studies of the P-gp substrates, saquinavir and paclitaxel, were performed to evaluate function. Treatment with either E2 or P4 resulted in a significant increase in P-gp protein levels in the NCI-ADR-RES cells at concentrations of or greater than 100 nM or 10 nM, respectively. JAR cells also had increased levels of P-gp with 100 nM of P4 but were much more sensitive to E2 showing increased P gp at a concentration of 1 nM. Furthermore, E2 or P4 treatment resulted in a significant decrease in cellular uptake of the P-gp substrates tested in these cells lines. Based on mRNA quantitation, a transient increase (2-fold) in MDR1 levels was observed at 8 h postincubation with either E2 or P4, while MDR1 levels remained high in the JAR cells treated with E2 for 72 h postincubation. The addition of actinomycin D, a transcription inhibitor negated the increase in P-gp by P4 and E2. P4 and E2 increase P-gp expression and function in NCI-ADR-RES and JAR cells with the ERalpha-expressing cells (JAR) much more sensitive to E2. Furthermore, transcriptional regulation by E2 and P4 likely contributes to the modulation of P-gp levels. PMID- 19813764 TI - Examination of the role of Taft-type steric parameters in asymmetric catalysis. AB - We report the use of Taft steric parameters to correlate the substituent size of a ligand with the enantiomeric ratio of a reaction. Linear free energy relationships can be constructed by plotting the log of enantiomeric ratio (er) versus the steric parameters reported by Taft and modified by Charton. Successful correlations were found for aldehyde and ketone allylation under NHK conditions using modular oxazoline ligands developed in our laboratory. Using these correlations, ligands were designed and evaluated for carbonyl allylation reactions. A break in the Charton plot results and is attributed to a global structural change in the catalyst. Additionally, several previously reported enantioselective reactions are analyzed resulting in excellent correlations for both catalysts and substrates. Finally, limitations and issues are presented with illustrative examples. PMID- 19813765 TI - InBr3: a versatile catalyst for the different types of Friedel-Crafts reactions. AB - Mild and efficient InBr(3)-catalyzed Friedel-Crafts alkylation of heteroaromatic or electron-rich aromatic compounds with alpha-amido sulfones at room temperature in CH(2)Cl(2) has been developed. The products undergo further Friedel-Crafts alkylation with heteroaromatic or electron-rich aromatic compounds leading to unsymmetrical or bis-symmetrical triaryl methanes in good yield. Alpha-amido sulfones are employed for the synthesis of the unsymmetrical and bis-symmetrical triaryl methanes. The use of mild reaction condition, low catalytic loading, and high yield are the advantages of the present procedures. PMID- 19813766 TI - Olefin metathesis-iodoetherification-dehydroiodination strategy for spiroketal subunits of polyether antibiotics. AB - The convergent synthesis of two pentacyclic analogues of the polyether monensin A is described. Although different with respect to the configuration of the alcohol at the 3 position of the six-membered ring of the spiroketal subunit, the configuration at the acetal center in both structures is unchanged and is consistent with the anomeric effect. The key synthetic steps are the coupling of two complex segments via an olefin metathesis, and the subsequent conversion of a dihydroxyalkene to the spiroketal through an iodoetherification-dehydroiodination sequence. The compatibility of these transformations with a variety of functional groups makes the overall strategy appropriate for highly substituted frameworks. PMID- 19813767 TI - Synthesis of alpha-amino amides via N,O-acetals derived from Weinreb amides. AB - An easy and straightforward synthesis of alpha-amino amides via a base-mediated rearrangement of modified Weinreb amides into N,O-acetals is presented. Subsequent arylation, alkylation, alkenylation, or alkynylation of this intermediate affords the corresponding alpha-amino amides in excellent yields. Furthermore, a more generalized protocol for the alpha-arylation of Weinreb amides lacking an alpha-amino moiety is also discussed. PMID- 19813768 TI - Studies on the synthesis of the ABC rings of (+/-)-hexacyclinic acid. AB - A synthesis of the ABC-rings of the polyketide natural product hexacyclinic acid has been achieved. The B-ring was formed first via an intramolecular ester tethered Diels-Alder reaction, and the A-ring was annulated to this by means of a SmI(2) mediated reductive 5-exo-trig cyclization of a samarium-ketyl radical onto a vinyl group. Finally, the C-ring was closed using olefin metathesis. Interestingly, use of enyne metathesis resulted in the synthesis of a more functionalized 5-membered C-ring in a model system, but an undesired 6-membered C ring in the actual system. An investigation of this change in selectivity is discussed. PMID- 19813769 TI - Synthesis of hybrid masked triyne-phenylene axial rods containing (E)-beta chlorovinylsilanes in the pi-conjugated framework. AB - A two-directional synthesis of a masked hexayne 7, in which two beta chlorovinylsilanes protect two of the internal alkynes, is reported. The key step involves the Pd-catalyzed oxidative dimerization of alkyne 10 to provide diyne 12, which is elaborated into centrosymmetric masked hexayne 7 in four steps. Masked hexayne 7 is a constitutional isomer of masked hexayne 2, which has been used as a monomer unit for oligoyne assembly. Although masked hexayne 7 was not as convenient a building block as 2 for application in oligoyne assembly, one of its precursors, namely alkyne 10, could be used successfully in Sonogashira couplings, which allowed the incorporation of aromatic spacers and the formation of hybrid masked triyne-phenylenes 20 and 28. Compounds 20 and 28 both contain removable end-groups, which will permit their application as building blocks for the assembly of classes of long-chain, pi-conjugated rod-like molecules. Rod-like molecule 34, which possesses a similar conjugated scaffold as 28, was also prepared by using a similar strategy. Treatment of 34 with TBAF effected a 2-fold dechlorosilylation to provide a rigid rod molecule 35 in which two phenylene units interrupt an octayne scaffold. PMID- 19813770 TI - Improving metabolite knowledge in stable atherosclerosis patients by association and correlation of GC-MS and 1H NMR fingerprints. AB - The plasma of patients with stable carotid atherosclerosis (n = 9), and healthy subjects (n = 10) have been fingerprinted with both GC-MS and (1)H NMR. Principal component analysis (PCA), partial least-squares-discriminant analysis (PLS-DA) and orthogonal partial least-squares-discriminant analysis (OPLS-DA) have been applied to the profiles from each technique both separately and in combination. These techniques complement each other and enable a clearer picture of the biological samples to be interpreted not only for classification purposes, but also more importantly to define the metabolic state of patients with carotid atherosclerosis. The results showed at least 24 metabolites that were significantly modified in the group of atherosclerotic patients by this nontargeted procedure. Most of the changes can be associated to alterations of the metabolism characteristics of insulin resistance that can be strongly related to the metabolic syndrome. In addition, correlations among variables accounting for the classification show amino acids as variables whose changes showed a high degree of correlation. GC-MS and (1)H NMR fingerprints can provide complementary information in the identification of altered metabolic pathways in patients with carotid atherosclerosis. Moreover, correlations among the results with both techniques, instead of a single study, can provide a deeper insight into the patient state. PMID- 19813771 TI - A dynamic range compression and three-dimensional peptide fractionation analysis platform expands proteome coverage and the diagnostic potential of whole saliva. AB - Comprehensive identification of proteins in whole human saliva is critical for appreciating its full diagnostic potential. However, this is challenged by the large dynamic range of protein abundance within the fluid. To address this problem, we used an analysis platform that coupled hexapeptide libraries for dynamic range compression (DRC) with three-dimensional (3D) peptide fractionation. Our approach identified 2340 proteins in whole saliva and represents the largest saliva proteomic dataset generated using a single analysis platform. Three-dimensional peptide fractionation involving sequential steps of preparative isoelectric focusing (IEF), strong cation exchange, and capillary reversed-phase liquid chromatography was essential for maximizing gains from DRC. Compared to saliva not treated with hexapeptide libraries, DRC substantially increased identified proteins across physicochemical and functional categories. Approximately 20% of total salivary proteins are also seen in plasma, and proteins in both fluids show comparable functional diversity and disease-linkage. However, for a subset of diseases, saliva has higher apparent diagnostic potential. These results expand the potential for whole saliva in health monitoring/diagnostics and provide a general platform for improving proteomic coverage of complex biological samples. PMID- 19813772 TI - Clinical and economic comparison of frovatriptan versus other oral triptans in the treatment of acute migraine in the real-world setting. AB - Triptans (serotonin 5-HT(1B/1D) receptor agonists) such as frovatriptan have been shown to be highly effective and well tolerated in the treatment of patients with acute migraine. However, the large number of available triptans has led to the issue of how best to decide which triptan should be prescribed at an individual patient level. This review focuses on frovatriptan and highlights parameters that affect oral triptan choice, discusses the results of several open-label clinical and post-marketing surveillance studies of frovatriptan, and compares these naturalistic data with those from similar studies of other oral triptans. Efficacy data obtained from these trials are used to compare costs of treating migraine with frovatriptan and other oral triptans in four European countries. Studies of triptans in migraine have used several outcomes deemed important to patients, including complete pain relief, absence of recurrence, rapid onset of action, no side effects, restoration of functional ability, improvements in quality of life, and cost. In contrast to indirect evidence from some individual randomized, double-blind studies, results from open-label naturalistic studies and a meta-analysis of 31 placebo-controlled efficacy studies suggest that frovatriptan is associated with a lower rate of migraine recurrence than with other triptans in a real-world clinical setting (17% for frovatriptan 2.5 mg vs 23-40% for other triptans in the meta-analysis). It is likely that this may be due to the terminal elimination half-life of this agent (about 26 hours), which is longer than that of other triptans. Naturalistic studies indicate that the long duration of action of frovatriptan appears to confer other benefits such as greater patient satisfaction, with over 90% of patients and doctors rating frovatriptan therapy as 'very good' or 'good'. The cost of treatment with different triptans based on the number of tablets required per episode varies widely in each of the four countries analysed (Great Britain Pound 4.95-7.98 in France, Great Britain Pound 6.78-12.58 in Germany, Great Britain Pound 4.32-9.73 in the UK and Great Britain Pound 6.69-10.36 in Italy, based on lowest possible costs for branded versions in 2008) due to differences in both the acquisition costs of these agents and in the headache recurrence rates. Frovatriptan compares favourably with other available triptans with regard to cost per migraine attack on this basis, although head-to-head studies are required to confirm these data. The low rate of headache recurrence with frovatriptan compared with other oral triptans, and the associated lower treatment costs, deserve consideration when choosing an oral triptan for the treatment of patients with acute migraine. PMID- 19813773 TI - Perioperative use of nonsteroidal anti-inflammatory drugs: results of a UK regional audit. AB - BACKGROUND AND OBJECTIVE: The use of nonsteroidal anti-inflammatory drugs (NSAIDs) is well established as a part of multimodal perioperative analgesia treatment, especially in day-case surgery where opioid sparing is important. The aim of this study was to assess the current perioperative use of NSAIDs, the specific drugs used, and the route by which they were administered. METHODS: A pilot study was undertaken, followed by a second, larger study. Numbered questionnaires were sent to all members of the anaesthetic departments of six target National Health Service Hospitals within the south of the UK. The questionnaires asked specific questions relating to the use of NSAIDs in their departments during the preoperative, perioperative and postoperative periods. Responses to the questionnaires were anonymous except for the identity of the hospital concerned. RESULTS: The pilot study indicated that perioperative NSAIDs continue to be used by anaesthetists and this was confirmed by the second audit. In total, 371 questionnaires were sent out, with 51% of recipients (189/371) responding. All of the respondents reported using NSAIDs (diclofenac, ibuprofen, ketorolac, ketoprofen and parecoxib), with many of the respondents using more than one NSAID. Diclofenac was the most commonly used NSAID overall. However, some of the NSAIDs - notably ketorolac (41% of respondents), diclofenac (61% of respondents) and ketoprofen (7% of respondents) were not used according to the manufacturer's recommendations. CONCLUSION: These data reveal that NSAIDs were still widely used as analgesics for many surgical procedures at the time of the audit. Intravenous (IV) NSAID administration is the preferred route because of its reliability and speed of onset. The results of this audit indicated significant use of IV NSAIDs (ketorolac, diclofenac and ketoprofen) not in accordance with manufacturers' recommendations, with some NSAIDs even being used in the absence of a product licence for use by the IV route. This may be due to a lack of a satisfactory licensed product. A new formulation of IV diclofenac may fulfil this currently unmet need. PMID- 19813774 TI - Pharmacokinetics and antihypertensive effects of candesartan cilexetil in patients undergoing haemodialysis: an open-label, single-centre study. AB - BACKGROUND AND OBJECTIVE: In patients with endstage renal failure (ERF), activation of the renin-angiotensin-aldosterone system plays an important role in the onset and maintenance of arterial hypertension. This study aimed to elucidate the antihypertensive effect, pharmacokinetics and safety of candesartan cilexetil in patients with ERF undergoing haemodialysis. METHODS: In 14 anuric hypertensive patients undergoing haemodialysis (mean+/- SD 24-hour systolic [SBP]/diastolic [DBP] blood pressure [BP] 142.9 +/- 11.1/75.0 +/- 10.1 mmHg), 24-hour BP measurements on the second interdialysis day per week were performed at baseline and at weeks 4, 12 and 24. All patients started antihypertensive treatment with candesartan cilexetil 4 mg once daily immediately before the start of haemodialysis. Subsequently, the dose was titrated upward to 8 mg once daily until the patient's mean ambulatory BP measurement (ABPM) values were <130/80 mmHg. Plasma candesartan pharmacokinetics were investigated on days 7 and 14 after starting candesartan cilexetil treatment and after each titration step. RESULTS: After 6 months all patients demonstrated well controlled BP (ABPM mean +/- SD SBP 129.6 +/- 21.7/DBP 69.4 +/- 10.4 mmHg) and a significantly reduced pulse pressure (from a mean +/- SD 67.9 +/- 13.7 mmHg at baseline to a mean +/- SD 60.2 +/- 14.7 mmHg at 6 months), without any adverse events. Candesartan plasma concentrations increased over 3 hours followed by a continuous decline. Plasma concentrations remained stable after 7 and 14 days, independent of dosing. However, administration of candesartan cilexetil 8 mg (five patients) resulted in plasma concentrations about 1.4 times higher than those for candesartan cilexetil 4 mg. CONCLUSION: In this study with small number of patients with ERF undergoing haemodialysis, candesartan cilexetil was effective in lowering BP and pulse pressure without accumulation or associated adverse effects such as elevated potassium or symptomatic hypotension. PMID- 19813775 TI - Comparison of the pharmacokinetics of apricitabine in the presence and absence of ritonavir-boosted tipranavir: a phase I, open-label, controlled, single-centre study. AB - BACKGROUND AND OBJECTIVE: Apricitabine is a deoxycytidine analogue nucleoside reverse transcriptase inhibitor for the treatment of HIV infection. The aim of this phase I study was to investigate whether administration of apricitabine with the HIV protease inhibitor tipranavir (ritonavir-boosted) affects the pharmacokinetic profile of apricitabine. METHODS: This phase I study was conducted in 18 healthy adult male subjects. Subjects received a single dose of apricitabine 800 mg on the morning of day 1 followed by tipranavir 500 mg plus ritonavir 200 mg every 12 hours from day 2 to day 9 to achieve steady-state concentrations of tipranavir/ritonavir. On day 10, subjects received a single morning dose of apricitabine 800 mg and a single dose of tipranavir 500 mg plus ritonavir 200 mg. Following dosing on days 1, 9 and 10, pharmacokinetic sampling was undertaken over 12 hours post-dosing to determine the plasma concentrations of apricitabine and tipranavir. RESULTS: The administration of a single dose of apricitabine 800 mg in the presence of steady-state tipranavir/ritonavir concentrations resulted in an increase in the apricitabine area under the plasma concentration-time curve of approximately 40% and in the apricitabine maximum plasma concentration of approximately 25% relative to apricitabine 800 mg administered alone. Apricitabine was well tolerated when administered with tipranavir/ritonavir. CONCLUSION: A moderate increase in apricitabine exposure was seen after co-administration with ritonavir-boosted tipranavir but this increase was not of clinical significance. No adjustment of apricitabine dosing is required when administered with ritonavir-boosted tipranavir. PMID- 19813776 TI - Safety/Tolerability and efficacy of rivastigmine in taiwanese patients with Alzheimer's disease: a prospective post-marketing surveillance study. AB - BACKGROUND AND OBJECTIVES: Rivastigmine is approved for the symptomatic treatment of mild to moderate dementia in patients with Alzheimer's disease. The drug was launched in Taiwan in 2000. The primary objective of this post-marketing surveillance (PMS) study was to describe the safety/tolerability of treatment with rivastigmine capsules in patients with Alzheimer's disease. The secondary objectives of this study were to define the optimal titration pattern, maintenance dose, efficacy and patient satisfaction with treatment with rivastigmine capsules. METHODS: This was a prospective, non-interventional post marketing observational study in patients who met the criteria for mild or moderate Alzheimer's disease. The primary outcome measure for this trial was the incidence of emerging adverse events. Dosages related to titration patterns and maintenance doses were summarized. Efficacy evaluations conducted using the Mini Mental State Examination, Clinical Dementia Rating and modified Instrumental Activities of Daily Living scales were also primary outcome measures, and results are shown descriptively. The patients' therapeutic responses to rivastigmine and satisfaction with rivastigmine were secondary outcome measures. Therapeutic response and treatment satisfaction were summarized descriptively. RESULTS: A total of 264 patients were enrolled into the study. The mean duration of exposure to rivastigmine during the study was 151.1 days. Patients were taking rivastigmine 1.5-6 mg twice daily and the most frequent maintenance dose level was 4.5 mg twice daily. Among patients treated with rivastigmine, all primary and secondary outcome measures showed improvement or stabilization of cognition and global functioning. Of the 253 safety analysis patients, 155 patients (61.3%) reported at least one adverse event. The most frequent adverse events by system organ class were psychiatric disorders (9.1%) and gastrointestinal disorders (8.3%). The most common adverse events observed were dizziness (5.5%), insomnia (5.1%), anorexia (4.0%) and gastrointestinal symptoms such as constipation (4.0%), vomiting (4.0%) and nausea (3.6%). These symptoms were mild in severity. A total of 12 patients (4.7%) reported 16 serious adverse events, including two deaths, three fractures, three behavioural and psychological symptoms of dementia, one syncope with head trauma, one peptic ulcer, and six other hospitalizations. None were reported to be related to rivastigmine. CONCLUSIONS: Based on the results of this study, rivastigmine administered usually at a dose of 3-6 mg twice daily was found to be well tolerated. Although the rate of adverse events was high, the majority of these symptoms were mild in severity and short in duration. This study also demonstrated the efficacy of rivastigmine in at least stabilizing the symptoms of Alzheimer's disease. PMID- 19813777 TI - Effects of benidipine, a long-acting T-type calcium channel blocker, on home blood pressure and renal function in patients with essential hypertension: a retrospective, 'real-world' comparison with amlodipine. AB - BACKGROUND AND OBJECTIVE: Calcium channel antagonists (calcium channel blockers [CCBs]) are often used in the treatment of patients with hypertension to achieve strict blood pressure (BP) targets. In the present study, we compared the antihypertensive effects (determined by home BP [HBP] measurements) and the effects on renal function of benidipine (hydrochloride) and amlodipine (mesylate), a commonly used CCB. METHODS: Changes in HBP and urinary albumin excretion (UAE) were investigated in 47 benidipine and 37 amlodipine recipients with essential hypertension and albuminuria between January 2007 and December 2007. Both benidipine and amlodipine significantly reduced morning and evening HBP over a 12-month period. RESULTS: Both medications also significantly reduced UAE compared with pretreatment values; however, the reduction in UAE observed in the benidipine group occurred independent of the drug's antihypertensive effects, whereas a positive correlation was shown between the reduction in morning systolic BP and UAE in the amlodipine group. CONCLUSIONS: These results demonstrate that benidipine favourably affects renal function in patients with essential hypertension compared with amlodipine, suggesting that the clinical benefits of benidipine as an antihypertensive drug include a renoprotective effect. PMID- 19813779 TI - On the sign of the adaptive passive fathometer impulse response. AB - Harrison [J. Acoust. Soc. Am. 125, 3511-3513 (2009)] presented a mathematical explanation for a sign-inversion induced to the passive fathometer response by minimum variance distortionless response (MVDR) beamforming. Here a concise mathematical formulation is offered, which decomposes the cross-spectral density matrix into coherent and incoherent components and allows the matrix inversion to be obtained exactly by eigendecomposition. This shows that, in the region containing the bottom reflection, the MVDR fathometer response is identical to that obtained with conventional processing multiplied by a negative factor. PMID- 19813778 TI - Pancytopenia associated with levetiracetam treatment. AB - Levetiracetam is a pyrrolidine derivate that is approved for the treatment of seizures. It has a favourable pharmacokinetic profile, no clearly established pharmacological interactions, good tolerability and offers the possibility of rapid treatment. We describe a case of pancytopenia and multiple infections observed in the context of levetiracetam treatment. A 65-year-old woman who underwent surgical removal of a meningioma developed progressive pancytopenia complicated by pneumonia and multiple liver abscesses after starting levetiracetam therapy. Bone marrow aspiration and biopsy confirmed the diagnosis. A gradual spontaneous recovery of normal haematopoiesis followed discontinuation of levetiracetam. PMID- 19813780 TI - Time domain visualization using acoustic holography implemented by temporal and spatial complex envelope. AB - Spatial envelope was proposed to show the location of acoustic sources and overall radiation pattern by the authors [C.-S. Park and Y.-H. Kim, J. Acoust. Soc. Am. 125, 206-211 (2009)]. The envelope can provide sufficient information on where the sources are and how the energy propagates into space. This concept is certainly useful for time domain acoustic holography since one can utilize not only spatial envelope to envisage what one needs to know but also temporal envelope to reduce the number of data. A holographic process to obtain spatial envelope is therefore introduced and verified, and how much one can reduce the processing time by implementing envelopes is compared with the conventional holography. PMID- 19813781 TI - Octave-shifted pitch matching in nonword imitations: the effects of lexical stress and speech sound disorder. AB - Perceptual similarities of musical tones separated by octave intervals are known as octave equivalence (OE). Peter et al. [(2008). Proceedings of the Fourth Conference on Speech Prosody, edited S. Maduerira, C. Reis, and P. Barbosa, Luso Brazilian Association of Speech Sciences, Campinas, pp. 731-734] found evidence of octave-shifted pitch matching (OSPM) in children during verbal imitation tasks, implying OE in speech tokens. This study evaluated the role of lexical stress and speech sound disorder (SSD) in OSPM. Eleven children with SSD and 11 controls imitated low-pitched nonwords. Stimulus/response f(0) ratios were computed. OSPM was expressed preferentially in stressed vowels. SSD was associated with reduced expression of OSPM in unstressed vowels only. Results are consistent with the psycholinguistic prominence of lexical stress and prosodic deficits in SSD. PMID- 19813782 TI - Acoustic standing wave suppression using randomized phase-shift-keying excitations. AB - Recent papers have demonstrated that acoustic standing waves can be inhibited by frequency-modulated spread-spectrum excitation. An alternative method is studied here that is designed to be more practical for implementation in phased arrays. The method operates using phase-shift-keying (PSK), which introduces phase shifts into the driving signal to break wave symmetry. Sequential and random binary-PSK (BPSK) and quadrature-PSK (QPSK) excitations are studied in water, using a carrier frequency of 250 kHz and a time segment of 10 cycles. The resulting acoustic field is measured with a transducer inside a plastic-walled chamber and compared with continuous wave excitation. Results indicate that both the random BPSK and QPSK methods can reduce time-averaged spatial intensity variation caused by standing waves by approximately six times. PMID- 19813783 TI - Comments on "The science and applications of acoustics" [Springer-Verlag (2000, 2006)]. AB - Raichel's "The Science and Applications of Acoustics" (Springer-Verlag, Berlin, 2000) has extensive material that appears to have been extracted from four other texts without proper attribution. The material so used extends to figures, detailed structure of entire sections and developments, and phrasing of individual passages. While in some instances like material between the texts has been paraphrased to some extent, in others it appears almost unaltered. PMID- 19813784 TI - Propagation of nonlinear acoustic plane waves in an elastic gas-filled tube. AB - This paper deals with modeling of nonlinear plane acoustic waves propagating through an elastic tube filled with thermoviscous gas. A description of the interactions between gas and an elastic tube wall is carried out by the continuity equation of a wall velocity. Simplification on the basis of the local reaction assumption enables to model an acoustic treatment on the tube wall by using a wall impedance. Because there are considerable losses due to wall friction, the influences of the acoustic boundary layer were also considered. Using certain assumptions a special form of the Burgers equation was derived which enables to describe the propagation of nonlinear waves in the elastic tube. This model equation takes into account nonlinear, dissipative, and dispersion effects which compete each other. Characteristic lengths of the supposed effects and numerical results with respect to the source frequency were used for a qualitative analysis of the model equation. Applicability of this model equation was demonstrated by series of measurements. By application of the long-wave approximation the Korteweg-de Vries-Burgers and Kuramoto-Sivashinsky equations were derived from the modified Burgers equation. PMID- 19813785 TI - Numerical investigation of nonlinear propagation distortion effects in helicopter rotor noise. AB - The effect of nonlinear propagation distortion on helicopter rotor noise is presented based on measured data for low-speed descent and numerical calculations that predict the noise level away from the helicopter with and without nonlinear effects. It is shown that for some frequency bands the difference between linear and nonlinear calculations can be as high as 7 dB. Blade vortex interaction (BVI) noise, the dominant noise contributor during descent, is mainly examined. It is shown that advancing side BVI noise is affected by nonlinear distortion, while retreating side BVI noise is not. Based on signal characteristics at source, two quantities are derived. The first quantity (termed polarity) is based on the pressure gradient of the source signal and can be used to determine whether a BVI signal will evolve as an advancing or a retreating side signal. The second quantity (termed weighted rise time) is a measure of the impulsiveness of the BVI signal and can be used to determine at which frequency nonlinear effects start to appear. Finally, polarity and weighted rise time are shown to be applicable in cases of BVI noise generated from different blade tips, as well as in cases of non-BVI noise. PMID- 19813786 TI - A three dimensional parabolic equation method for sound propagation in moving inhomogeneous media. AB - In this paper, a formulation of the Helmholtz equation for three dimensional sound propagation in a moving inhomogeneous medium in cylindrical coordinates is derived. Based on this formulation, a three dimensional parabolic equation (PE) is constructed. This PE can be used to model sound propagation in an inhomogeneous arbitrary moving medium. The method is used here to simulate three dimensional outdoor sound propagation above a rigid flat ground surface. The numerical results for two simple wind cases are presented and compared with analytical results to validate the methodology. Examples of propagation problems with more complicated wind are then included to demonstrate the importance of including the wind velocity directly in the PE method. PMID- 19813787 TI - Radiative transfer theory applied to ocean bottom modeling. AB - Research on the propagation of acoustic waves in the ocean bottom sediment is of interest for active sonar applications such as target detection and remote sensing. The interaction of acoustic energy with the sea floor sublayers is usually modeled with techniques based on the full solution of the wave equation, which sometimes leads to mathematically intractable problems. An alternative way to model wave propagation in layered media containing random scatterers is the radiative transfer (RT) formulation, which is a well established technique in the electromagnetics community and is based on the principle of conservation of energy. In this paper, the RT equation is used to model the backscattering of acoustic energy from a layered elastic bottom sediment containing distributions of independent scatterers due to a constant single frequency excitation in the water column. It is shown that the RT formulation provides insight into the physical phenomena of scattering and conversion of energy between waves of different polarizations. PMID- 19813788 TI - Riverbed sediment classification using multi-beam echo-sounder backscatter data. AB - A method has recently been developed that employs multi-beam echo-sounder backscatter data to both obtain the number of sediment classes and discriminate between them by applying the Bayes decision rule to multiple hypotheses [Simons and Snellen, Appl. Acoust. 70, 1258-1268 (2009)]. In deep water, the number of scatter pixels within the beam footprint is large enough to ensure Gaussian distributions for the backscatter strengths and to increase the discriminative power between acoustic classes. In very shallow water (<10 m), however, this number is too small. This paper presents an extension of this high-frequency methodology for these environments, together with a demonstration of its performance using backscatter data from the river Waal, The Netherlands. The objective of this work is threefold. (i) Increasing the discriminating power of the classification method: high-resolution bathymetry data allow precise bottom slope corrections for obtaining the true incident angle, and the high-resolution backscatter data reduce the statistical fluctuations via an averaging procedure. (ii) Performing a correlation analysis: the dependence of acoustic backscatter classification on sediment physical properties is verified by observing a significant correlation of 0.75 (and a disattenuated correlation of 0.90) between the classification results and sediment mean grain size. (iii) Enhancing the statistical description of the backscatter intensities: angular evolution of the K-distribution shape parameter indicates that the riverbed is a rough surface, in agreement with the results of the core analysis. PMID- 19813789 TI - On the consideration of motion effects in the computation of impulse response for underwater acoustics inversion. AB - The estimation of the impulse response (IR) of a propagation channel may be of great interest for a large number of underwater applications: underwater communications, sonar detection and localization, marine mammal monitoring, etc. It quantifies the distortions of the transmitted signal in the underwater channel and enables geoacoustic inversion. The propagating signal is usually subject to additional and undesirable distortions due to the motion of the transmitter channel-receiver configuration. This paper shows the effects of the motion while estimating the IR by matched filtering between the transmitted and the received signals. A methodology to compare IR estimation with and without motion is presented. Based on this comparison, a method for motion effect compensation is proposed in order to reduce motion-induced distortions. The proposed methodology is applied to real data sets collected in 2007 by the Service Hydrographique et Oceanographique de la Marine in a shallow water environment, proving its interest for motion effect analysis. Motion compensated estimation of IRs is computed from sources transmitting broadband linear frequency modulations moving at up to 12 knots in the shallow water environment of the Malta plateau, South of Sicilia. PMID- 19813790 TI - Acoustic mode radiation from the termination of a truncated nonlinear internal gravity wave duct in a shallow ocean area. AB - Horizontal ducting of sound between short-wavelength nonlinear internal gravity waves in coastal environments has been reported in many theoretical and experimental studies. Important consequences arising at the open end of an internal wave duct (the termination) are examined in this paper with three dimensional normal mode theory and parabolic approximation modeling. For an acoustic source located in such a duct and sufficiently far from the termination, some of the propagating sound may exit the duct by penetrating the waves at high grazing angles, but a fair amount of the sound energy is still trapped in the duct and propagates toward the termination. Analysis here shows that the across duct sound energy distribution at the termination is unique for each acoustic vertical mode, and as a result the sound radiating from the termination of the duct forms horizontal beams that are different for each mode. In addition to narrowband analysis, a broadband simulation is made for water depths of order 80 m and propagation distances of 24 km. Situations occur with one or more modes absent in the radiated field and with mode multipath in the impulse response. These are both consistent with field observations. PMID- 19813791 TI - Wave scattering from encapsulated microbubbles subject to high-frequency ultrasound: contribution of higher-order scattering modes. AB - The theoretical understanding of encapsulated microbubble response to high frequency ultrasound (HFUS) excitation is still limited although some novel experimental HFUS contrast imaging techniques have been well developed. In this paper, the higher-order modal (HOM) contributions to the scattered field are studied for such microbubbles driven by 1-100 MHz ultrasound. An exact solution of all small-amplitude vibrational modes of a single encapsulated microbubble in water is given by the wave scattering theory (WST) method and compared to results obtained from Church's Rayleigh-Plesset-like model for the small-amplitude radial oscillation of a microbubble in an incompressible fluid. From numerical results, we show that the HOM field contribution is significant for scattering properties from individual Nycomed microbubbles with normalized frequency > or = 0.2. It is also shown that the multiple scattering is strengthened for monodispersed Definity microbubbles of 3 microm radius at frequencies >40 MHz. However, comparisons between the authors' analyses and known experimental data for polydispersed Definity microbubbles indicate that the HOM contributions are insignificant in attenuation estimation at frequencies <50 MHz. In conclusion, the WST model analysis suggests that HOM scattering is an important consideration for single bubbles but may be less critical in the modeling of polydispersed Definity bubbles at high frequencies. PMID- 19813792 TI - Sound radiation quantities arising from a resilient circular radiator. AB - Power series expansions in ka are derived for the pressure at the edge of a radiator, the reaction force on the radiator, and the total radiated power arising from a harmonically excited, resilient, flat, circular radiator of radius a in an infinite baffle. The velocity profiles on the radiator are either Stenzel functions (1-(sigma/a)2)n, with sigma the radial coordinate on the radiator, or linear combinations of Zernike functions Pn(2(sigma/a)2-1), with Pn the Legendre polynomial of degree n. Both sets of functions give rise, via King's integral for the pressure, to integrals for the quantities of interest involving the product of two Bessel functions. These integrals have a power series expansion and allow an expression in terms of Bessel functions of the first kind and Struve functions. Consequently, many of the results in [M. Greenspan, J. Acoust. Soc. Am. 65, 608-621 (1979)] are generalized and treated in a unified manner. A foreseen application is for loudspeakers. The relation between the radiated power in the near-field on one hand and in the far field on the other is highlighted. PMID- 19813793 TI - Hybrid method for determining the parameters of condenser microphones from measured membrane velocities and numerical calculations. AB - Typically, numerical calculations of the pressure, free-field, and random incidence response of a condenser microphone are carried out on the basis of an assumed displacement distribution of the diaphragm of the microphone; the conventional assumption is that the displacement follows a Bessel function. This assumption is probably valid at frequencies below the resonance frequency. However, at higher frequencies the movement of the membrane is heavily coupled with the damping of the air film between membrane and backplate and with resonances in the back chamber of the microphone. A solution to this problem is to measure the velocity distribution of the membrane by means of a non-contact method, such as laser vibrometry. The measured velocity distribution can be used together with a numerical formulation such as the boundary element method for estimating the microphone response and other parameters, e.g., the acoustic center. In this work, such a hybrid method is presented and examined. The velocity distributions of a number of condenser microphones have been determined using a laser vibrometer, and these measured velocity distributions have been used for estimating microphone responses and other parameters. The agreement with experimental data is generally good. The method can be used as an alternative for validating the parameters of the microphones determined by classical calibration techniques. PMID- 19813794 TI - Channel separation of crosstalk cancellation systems with mismatched and misaligned sound sources. AB - Loudspeakers in virtual sound imaging systems are usually modeled as omnidirectional monopole sources. These models are, however, only an approximation for the low frequency range. This paper presents an analytical model of crosstalk cancellation systems in a free field which takes into account the scattering and spatial characteristics of the sound sources. Based on the proposed model, the effects caused by the spatial characteristics of the sound source and its misalignments on the performance of the crosstalk cancellation system are studied numerically. It is found that although the factors such as the directivity of the sound sources and the distance between the sound sources and receiver affect the performance of the system to a certain extent, the channel separation of the crosstalk cancellation system, however, is most sensitive to the misalignment of the subtended angle of the sound sources. Therefore, if highly accurate binaural cues are required in practical applications, the type and characteristics of the playback sound sources, their locations, and orientations all should be considered carefully. PMID- 19813795 TI - Steady state and time-dependent energy equilibration in two-dimensional random elastic slabs. AB - The static and dynamic transport properties of elastic wave propagation through two-dimensional random slabs without internal reflection were studied at two different scattering parameters: one for Rayleigh scattering and the other for Rayleigh-Gans scattering. The spatial distribution and temporal evolution of shear (s-) and compressional (p-) wave energy densities inside the slabs were calculated by solving the radiative transfer equation and the generalized diffusion equation (GDE). The comparison of their results can determine the region of validity of the GDE. The process of energy equilibration between the two wave modes was demonstrated explicitly as well as the process of diffusion. The depth inside a slab that is needed to reach energy equilibration or diffusive behavior is found to be dependent on source polarization. The results also show that the bulk equilibration ratio can be found inside a sample only when the sample is sufficiently thick. Deviations of the equilibration ratio from its bulk value are found near the output surface due to the absence of in-flow energy flux. The behavior of the deviations is sensitive to the scattering parameter but independent of source polarization. PMID- 19813796 TI - On the correlation of non-isotropically distributed ballistic scalar diffuse waves. AB - Theorems indicating that a fully equipartitioned random wave field will have correlations equivalent to the Green's function that would be obtained in an active measurement are now legion. Studies with seismic waves, ocean acoustics, and laboratory ultrasound have confirmed them. So motivated, seismologists have evaluated apparent seismic travel times in correlations of ambient seismic noise and tomographically constructed impressive maps of seismic wave velocity. Inasmuch as the random seismic waves used in these evaluations are usually not fully equipartitioned, it seems right to ask why it works so well, or even if the results are trustworthy. The error, in apparent travel time, due to non-isotropic specific intensity is evaluated here in a limit of large receiver-receiver separation and for the case in which the source of the noise is in the far field of both receivers. It is shown that the effect is small, even for cases in which one might have considered the anisotropy to be significant, and even for station pairs separated by as little as one or two wavelengths. A formula is derived that permits estimations of error and corrections to apparent travel time. It is successfully compared to errors seen in synthetic waveforms. PMID- 19813797 TI - Estimating sound power radiated from rectangular baffled panels using a radiation factor. AB - A method is introduced which is shown to predict radiated sound power from rectangular baffled panels. The method employs a filtered wavenumber transform to extract the power in the supersonic wavenumbers on the panel and a radiation factor to scale the supersonic power to match the actual radiated sound power. Although empirically derived, the radiation factor is shown to be related to the radiation efficiency of an infinite panel. The radiation factor is simple, depending only on the ratio of the wavenumbers of the panel to the radiation medium, and the method is straightforward to use, requiring only the panel normal velocities. The computation is efficient, as much as two orders of magnitude faster than a Rayleigh integration, thus providing a means of combining sound power predictions with finite element optimizations. A formula is derived which predicts the lowest frequency for which the method is valid as a function of the bin width of the wavenumber transform. The radiation factor method is shown to produce radiated sound power estimates which favorably compare to estimates derived from intensity measurements of physical test specimens and to Rayleigh integral estimates computed using both simulated and measured velocities. PMID- 19813798 TI - A comparative analysis of acoustic energy models for churches. AB - Different models to improve prediction of energy-based acoustic parameters in churches have been proposed by different researchers [E. Cirillo and F. Martellotta, J. Acoust. Soc. Am. 118, 232-248 (2005); T. Zamarreno et al., J. Acoust. Soc. Am. 121, 234-250 (2006)]. They all suggested variations to the "revised" theory proposed by Barron and Lee [J. Acoust. Soc. Am. 84, 618-628 (1988)], starting from experimental observations. The present paper compares these models and attempts to generalize their use taking advantage of the measurements carried out in 24 Italian churches differing in style, typology, and location. The whole sample of churches was divided into two groups. The first was used to fine-tune existing models, with particular reference to the "mu model," which was originally tested only on Mudejar-Gothic churches. Correlations between model parameters and major typological and architectural factors were found, leading to a classification that greatly simplifies parameter choice. Finally, the reliability of each model was verified on the rest of the sample, showing that acoustic parameters can be predicted with reasonable accuracy provided that one of the specifically modified theories is used. The results show that the model requiring more input parameters performs slightly better than the other which, conversely, is simpler to apply. PMID- 19813799 TI - Effect of boundary slip on the acoustical properties of microfibrous materials. AB - A variety of new porous materials with unusually small pores have been manufactured in the past decades. To predict their acoustical properties, the conventional models need to be modified. When pore size becomes comparable to the molecular mean free path of a saturating fluid, the no-slip conditions on the pore surface are no longer accurate and hence the slip effects have to be taken into account. In this paper, sound propagation in microfibrous materials is modeled analytically, approximating the geometry by a regular array of rigid parallel cylinders. It has been shown that velocity and thermal slip on a cylinder surface significantly changes the model predictions leading to lower attenuation coefficient and higher sound speed values. The influence of material porosity, fiber orientation, and size on these effects is investigated. Finite element method is used to numerically solve the oscillatory flow and heat transfer problems in a square array of cylindrical fibres. Numerical results are compared with predictions of the analytical model and the range of its validity is identified. PMID- 19813800 TI - Acoustic absorption calculation in irreducible porous media: a unified computational approach. AB - A critical task in predicting and tailoring the acoustic absorption properties of porous media is the calculation of the frequency-dependent effective density and compressibility tensors, which are explicitly related to the micro-scale permeability properties. Although these two quantities exhibit strong sensitivity to physics occurring at complex micro-scale geometries, most of the existing literature focuses on employing very limited in-house and oftentimes multiple numerical analysis tools. In order to predict these parameters and acoustic absorption efficiently and conveniently, this article synthesizes multiple disparate approaches into a single unified formulation suitable for incorporation into a commercial analysis package. Numerical results computed herein for four close-packed porous media are compared to similar results available in the literature. These include simple cubic, body-centered cubic, and face-centered cubic structures, and also hexagonal close-packed, which has not appeared in the literature. Together with critical comparisons of a hybrid versus direct numerical approaches, the close agreement demonstrates the capabilities of the unified formulation to analyze and control the acoustic absorption properties at the microscopic level. PMID- 19813801 TI - Predicting the sound insulation of single leaf walls: extension of Cremer's model. AB - In his 1942 paper on the sound insulation of single leaf walls, Cremer [(1942). Akust. Z. 7, 81-104] made a number of approximations in order to show the general trend of sound insulation above the critical frequency. Cremer realized that these approximations limited the application of his theory to frequencies greater than twice the critical frequency. This paper removes most of Cremer's approximations so that the revised theory can be used down to the critical frequency. The revised theory is used as a correction to the diffuse field limp panel mass law below the critical frequency by setting the nonexistent coincidence angle to 90 degrees. The diffuse field limp panel mass law for a finite size wall is derived without recourse to a limiting angle by following the average diffuse field single sided radiation efficiency approach. The shear wave correction derived by Heckl and Donner [(1985). Rundfunktech Mitt. 29, 287-291] is applied to the revised theory in order to cover the case of thicker walls. The revised theory predicts the general trend of the experimental data, although the agreement is usually worse at low frequencies and depends on the value of damping loss factor used in the region of and above the critical frequency. PMID- 19813802 TI - Relation between derived-band auditory brainstem response latencies and behavioral frequency selectivity. AB - Derived-band click-evoked auditory brainstem responses (ABRs) were obtained for normal-hearing (NH) and sensorineurally hearing-impaired (HI) listeners. The latencies extracted from these responses, as a function of derived-band center frequency and click level, served as objective estimates of cochlear response times. For the same listeners, auditory-filter bandwidths at 2 kHz were estimated using a behavioral notched-noise masking paradigm. Generally, shorter derived band latencies were observed for the HI than for the NH listeners. Only at low click sensation levels, prolonged latencies were obtained for some of the HI listeners. The behavioral auditory-filter bandwidths accounted for the across listener variability in the ABR latencies: Cochlear response time decreased with increasing filter bandwidth, consistent with linear-system theory. The results link cochlear response time and frequency selectivity in human listeners and offer a window to better understand how hearing impairment affects the spatiotemporal cochlear response pattern. PMID- 19813803 TI - Diotic and dichotic detection with reproducible chimeric stimuli. AB - Subject responses were measured for individual narrow-band reproducible stimuli in a low-frequency tone-in-noise detection task. Both N0S0 and N0Spi conditions were examined. The goal of the experiment was to determine the relative importance of envelope and fine-structure cues. Therefore, chimeric stimuli were generated by recombining envelopes and fine structures from different reproducible stimuli. Detection judgments for noise-alone or tone-plus-noise stimuli that had common envelopes but different fine structures or common fine structures but different envelopes were compared. The results showed similar patterns of responses to stimuli that shared envelopes, indicating the importance of envelope cues; however, fine-structure cues were also shown to be important. The relative weight assigned to envelope and fine-structure cues varied across subjects and across interaural conditions. The results also indicated that envelope and fine-structure information are not processed independently. Implications for monaural and binaural models of masking are discussed. PMID- 19813804 TI - An evaluation of models for diotic and dichotic detection in reproducible noises. AB - Several psychophysical models for masked detection were evaluated using reproducible noises. The data were hit and false-alarm rates from three psychophysical studies of detection of 500-Hz tones in reproducible noise under diotic (N0S0) and dichotic (N0Spi) conditions with four stimulus bandwidths (50, 100, 115, and 2900 Hz). Diotic data were best predicted by an energy-based multiple-detector model that linearly combined stimulus energies at the outputs of several critical-band filters. The tone-plus-noise trials in the dichotic data were best predicted by models that linearly combined either the average values or the standard deviations of interaural time and level differences; however, these models offered no predictions for noise-alone responses. The decision variables of more complicated temporal models, including the models of Dau et al. [(1996a). J. Acoust. Soc. Am. 99, 3615-3622] and Breebaart et al. [(2001a). J. Acoust. Soc. Am. 110, 1074-1088], were weakly correlated with subjects' responses. Comparisons of the dependencies of each model on envelope and fine-structure cues to those in the data suggested that dependence upon both envelope and fine structure, as well as an interaction between them, is required to predict the detection results. PMID- 19813805 TI - Informational masking in young and elderly listeners for speech masked by simultaneous speech and noise. AB - Three experiments measured the effects of age on informational masking of speech by competing speech. The experiments were designed to minimize the energetic contributions of the competing speech so that informational masking could be measured with no large corrections for energetic masking. Experiment 1 used a "speech-in-speech-in-noise" design, in which the competing speech was presented in noise at a signal-to-noise ratio (SNR) of -4 dB. This ensured that the noise primarily contributed the energetic masking but the competing speech contributed the informational masking. Equal amounts of informational masking (3 dB) were observed for young and elderly listeners, although less was found for hearing impaired listeners. Experiment 2 tested a range of SNRs in this design and showed that informational masking increased with SNR up to about an SNR of -4 dB, but decreased thereafter. Experiment 3 further reduced the energetic contribution of the competing speech by filtering it into different frequency bands from the target speech. The elderly listeners again showed approximately the same amount of informational masking (4-5 dB), although some elderly listeners had particular difficulty understanding these stimuli in any condition. On the whole, these results suggest that young and elderly listeners were equally susceptible to informational masking. PMID- 19813807 TI - Detection probability of vocalizing dugongs during playback of conspecific calls. AB - Dugongs (Dugong dugon) were monitored using simultaneous passive acoustic methods and visual observations in Thai waters during January 2008. Chirp and trill calls were detected by a towed stereo hydrophone array system. Two teams of experienced observers conducted standard visual observations on the same boat. Comparisons of detection probabilities of acoustic and visual monitoring between two independent observers were calculated. Acoustic and visual detection probabilities were 15.1% and 15.7%, respectively, employing a 300 s matching time interval. When conspecific chirp calls were broadcast from an underwater speaker deployed on the side of the observation boat, the detection probability of acoustic monitoring rose to 19.2%. The visual detection probability was 12.5%. Vocal hot spots characterized by frequent acoustic detection of calls were suggested by dispersion analysis, while dugongs were visually observed constantly throughout the focal area (p<0.001). Passive acoustic monitoring assisted the survey since detection performance similar to that of experienced visual observers was shown. Playback of conspecific chirps appeared to increase the detection probability, which could be beneficial for future field surveys using passive acoustics in order to ensure the attendance of dugongs in the focal area. PMID- 19813806 TI - Effects of external noise on detection of intensity increments. AB - The detection of an intensity increment in a longer duration sinusoid or pedestal is often used as a measure of intensity resolution, but the decision processes underlying this measure are poorly understood. Thresholds were obtained for detection of an increment in a 370-ms, 4-kHz pedestal in quiet or in noise to determine the relative contributions of background noise level and pedestal level, the effect of increment duration, and the effect of different noise spectra. Increment detection thresholds expressed in units of DeltaL[10 log(1+DeltaI/I)] decreased as pedestal levels increased. At low pedestal levels, increment detection was limited by the masking effect of the noise and was similar across noise conditions for pedestals of equal sensation level. At high pedestal levels, the noise had no effect and increment detection was determined by the pedestal level in dB SPL (sound pressure level). Increment detection improved with increasing increment duration and was altered less by a noise band above the pedestal/increment frequency than by a broadband noise that produced equal masking at the pedestal/increment frequency. The quadratic-compression model described by Neely and Jesteadt [(2005). Acta Acust. Acust. 91, 980-991] provided a better approximation to the data than a model based on excitation patterns. PMID- 19813809 TI - Auditory stream segregation in cochlear implant listeners: measures based on temporal discrimination and interleaved melody recognition. AB - The evidence that cochlear implant listeners routinely experience stream segregation is limited and equivocal. Streaming in these listeners was explored using tone sequences matched to the center frequencies of the implant's 22 electrodes. Experiment 1 measured temporal discrimination for short (ABA triplet) and longer (12 AB cycles) sequences (tone/silence durations = 60/40 ms). Tone A stimulated electrode 11; tone B stimulated one of 14 electrodes. On each trial, one sequence remained isochronous, and tone B was delayed in the other; listeners had to identify the anisochronous interval. The delay was introduced in the second half of the longer sequences. Prior build-up of streaming should cause thresholds to rise more steeply with increasing electrode separation, but no interaction with sequence length was found. Experiment 2 required listeners to identify which of two target sequences was present when interleaved with distractors (tone/silence durations = 120/80 ms). Accuracy was high for isolated targets, but most listeners performed near chance when loudness-matched distractors were added, even when remote from the target. Only a substantial reduction in distractor level improved performance, and this effect did not interact with target-distractor separation. These results indicate that implantees often do not achieve stream segregation, even in relatively unchallenging tasks. PMID- 19813808 TI - Integration of auditory and vibrotactile stimuli: effects of phase and stimulus onset asynchrony. AB - The perceptual integration of 250 Hz, 500 ms vibrotactile and auditory tones was studied in detection experiments as a function of (1) relative phase and (2) temporal asynchrony of the tone pulses. Vibrotactile stimuli were delivered through a single-channel vibrator to the left middle fingertip and auditory stimuli were presented diotically through headphones in a background of 50 dB sound pressure level broadband noise. The vibrotactile and auditory stimulus levels used each yielded 63%-77%-correct unimodal detection performance in a 2-I, 2-AFC task. Results for combined vibrotactile and auditory detection indicated that (1) performance improved for synchronous presentation, (2) performance was not affected by the relative phase of the auditory and tactile sinusoidal stimuli, and (3) performance for non-overlapping stimuli improved only if the tactile stimulus preceded the auditory. The results are generally more consistent with a "Pythagorean Sum" model than with either an "Algebraic Sum" or an "Optimal Single-Channel" Model of perceptual integration. Thus, certain combinations of auditory and tactile signals result in significant integrative effects. The lack of phase effect suggests an envelope rather than fine-structure operation for integration. The effects of asynchronous presentation of the auditory and tactile stimuli are consistent with time constants deduced from single-modality masking experiments. PMID- 19813810 TI - Simultaneous measures of electropalatography and intraoral pressure in selected voiceless lingual consonants and consonant sequences of German. AB - This work assessed relationships among intraoral pressure (IOP), electropalatographic (EPG) measures, and consonant sequence duration, in the following obstruents, clusters, and affricates of German: /t/, /sh/, /sht/, and /tsh/. The data showed significant correlations between IOP and percentage of articulatory contact (PC) for all speakers, whereas duration and place of articulation (measured by the EPG center of gravity) contributed less to IOP changes. Speakers differed in the strength of this relationship, possibly reflecting differences in vocal tract morphology or degree of laryngeal abduction. Single-point EPG and IOP measures in fricatives showed consistent correspondences across consonantal contexts, but the relationships for the stops were more complex and reflected positional effects. Temporal compression was observed for both members of the cluster, but only the fricative portion of the affricate. Conversely, coarticulation was observed for both the stop and fricative portion of the affricate, but only for the stop portion of the cluster, possibly reflecting biomechanical constraints. No clear differences were observed in coarticulatory resistance for stops and fricatives. These data contribute to a limited literature on articulatory-aerodynamic relationships in voiceless consonants and consonant sequences, and will provide a baseline for considering longer combinations of obstruents. PMID- 19813811 TI - Ranking vocal fold model parameters by their influence on modal frequencies. AB - The purpose of this study was to identify, using computational models, the vocal fold parameters which are most influential in determining the vibratory characteristics of the vocal folds. The sensitivities of vocal folds modal frequencies to variations model parameters were used to determine the most influential parameters. A detailed finite element model of the human vocal fold was created. The model was defined by eight geometric and six material parameters. The model included transitional boundary regions to idealize the complex physiological structure of real human subjects. Parameters were simultaneously varied over ranges representative of actual human vocal folds. Three separate statistical analysis techniques were used to identify the most and least sensitive model parameters with respect to modal frequency. The results from all three methods consistently suggest that a set of five parameters are most influential in determining the vibratory characteristics of the vocal folds. PMID- 19813812 TI - Acoustic-articulatory mapping in vowels by locally weighted regression. AB - A method for mapping between simultaneously measured articulatory and acoustic data is proposed. The method uses principal components analysis on the articulatory and acoustic variables, and mapping between the domains by locally weighted linear regression, or loess [Cleveland, W. S. (1979). J. Am. Stat. Assoc. 74, 829-836]. The latter method permits local variation in the slopes of the linear regression, assuming that the function being approximated is smooth. The methodology is applied to vowels of four speakers in the Wisconsin X-ray Microbeam Speech Production Database, with formant analysis. Results are examined in terms of (1) examples of forward (articulation-to-acoustics) mappings and inverse mappings, (2) distributions of local slopes and constants, (3) examples of correlations among slopes and constants, (4) root-mean-square error, and (5) sensitivity of formant frequencies to articulatory change. It is shown that the results are qualitatively correct and that loess performs better than global regression. The forward mappings show different root-mean-square error properties than the inverse mappings indicating that this method is better suited for the forward mappings than the inverse mappings, at least for the data chosen for the current study. Some preliminary results on sensitivity of the first two formant frequencies to the two most important articulatory principal components are presented. PMID- 19813813 TI - A biomechanical model of cardinal vowel production: muscle activations and the impact of gravity on tongue positioning. AB - A three-dimensional (3D) biomechanical model of the tongue and the oral cavity, controlled by a functional model of muscle force generation (lambda-model of the equilibrium point hypothesis) and coupled with an acoustic model, was exploited to study the activation of the tongue and mouth floor muscles during the production of French cardinal vowels. The selection of the motor commands to control the tongue and the mouth floor muscles was based on literature data, such as electromyographic, electropalatographic, and cineradiographic data. The tongue shapes were also compared to data obtained from the speaker used to build the model. 3D modeling offered the opportunity to investigate the role of the transversalis, in particular, its involvement in the production of high front vowels. It was found, with this model, to be indirect via reflex mechanisms due to the activation of surrounding muscles, not voluntary. For vowel /i/, local motor command variations for the main tongue muscles revealed a non-negligible modification of the alveolar groove in contradiction to the saturation effect hypothesis, due to the role of the anterior genioglossus. Finally, the impact of subject position (supine or upright) on the production of French cardinal vowels was explored and found to be negligible. PMID- 19813814 TI - Talker-to-listener distance effects on speech production and perception. AB - Simulating talker-to-listener distance (TLD) in virtual audio environments requires mimicking natural changes in vocal effort. Studies have identified several acoustic parameters manipulated by talkers when varying vocal effort. However, no systematic study has investigated vocal effort variations due to TLD, under natural conditions, and their perceptual consequences. This work examined the feasibility of varying the vocal effort cues for TLD in synthesized speech and real speech by (a) recording and analyzing single word tokens spoken at 1 m < or = TLD < or = 32 m, (b) creating synthetic and modified speech tokens that vary in one or more acoustic parameters associated with vocal effort, and (c) conducting perceptual tests on the reference, synthetic, and modified tokens to identify salient cues for TLD perception. Measured changes in fundamental frequency, intensity, and formant frequencies of the reference tokens across TLD were similar to other reports in the literature. Perceptual experiments that asked listeners to estimate TLD showed that TLD estimation is most accurate with real speech; however, large standard deviations in the responses suggest that reliable judgments can only be made for gross changes in TLD. PMID- 19813815 TI - Perceived loudness of speech based on the characteristics of glottal excitation source. AB - The impulse-like characteristic of glottal excitation in speech production is an important factor in the perception of loudness of speech signals. This characteristic is attributed to the abruptness of the closing phase in the glottal cycle. In this paper, an acoustic feature, called strength of excitation, is proposed to represent the impulse-like nature of excitation. The strength of excitation is derived from the linear prediction residual of speech signals, where the residual can be considered as an estimate of the source of excitation. Since the loudness of speech is perceived over one or more utterances of speech, it is hypothesized that the distribution of strength of excitation is indicative of the perceived loudness of speech. The distribution of strength of excitation is shown to distinguish between soft and loud utterances of speakers. The distribution can also help in discriminating between the loudness of two speakers. The loudness measure obtained using the distribution of the strength of excitation is in agreement with the subjective judgment of loudness of speech. PMID- 19813816 TI - Fractal ladder models and power law wave equations. AB - The ultrasonic attenuation coefficient in mammalian tissue is approximated by a frequency-dependent power law for frequencies less than 100 MHz. To describe this power law behavior in soft tissue, a hierarchical fractal network model is proposed. The viscoelastic and self-similar properties of tissue are captured by a constitutive equation based on a lumped parameter infinite-ladder topology involving alternating springs and dashpots. In the low-frequency limit, this ladder network yields a stress-strain constitutive equation with a time fractional derivative. By combining this constitutive equation with linearized conservation principles and an adiabatic equation of state, a fractional partial differential equation that describes power law attenuation is derived. The resulting attenuation coefficient is a power law with exponent ranging between 1 and 2, while the phase velocity is in agreement with the Kramers-Kronig relations. The fractal ladder model is compared to published attenuation coefficient data, thus providing equivalent lumped parameters. PMID- 19813817 TI - Why do Chinese alligators (Alligator sinensis) form bellowing choruses: a playback approach. AB - Crocodilians are quite vocal relative to other reptile groups, and the alligators are among the most vocal of the crocodilians. The Chinese alligator, Alligator sinensis, is usually solitary but engages in bellowing choruses in certain waters during the mating season. This paper reports the organization of Chinese alligator's bellowing choruses based upon field observations and playback experiments. Alligators of both genders engaged in the choruses, remaining immobile throughout and inclining toward bellowing synchronously (i.e., starting and finishing at about the same time). The choruses lasted about 10 min with abrupt onset and offset. Moreover, playback experiments revealed that both male and female alligators responded equally to bellowing stimuli from the same and opposite sexes and that none of the tested alligators approached the loudspeaker in spite of playback of male or female stimuli. These suggest that Chinese alligators may not bellow to compete for or attract mates during the choruses. Instead, when their ecological behaviors, namely, dispersed inhabitation, multi copulation, restricted mating season, etc., are considered, we hypothesize that they may synchronize bellows to enhance group detectability for assembling individuals into certain waters for subsequent copulations. PMID- 19813818 TI - Note types and coding in Parid vocalizations: the chick-a-dee call of the chestnut-backed chickadee (Poecile rufuscens). AB - A first step to understanding how a species communicates acoustically is to identify, categorize, and quantify the acoustic parameters of the elements that make up their vocalizations. The "chick-a-dee" call notes of the chestnut-backed chickadee (Poecile rufescens) were sorted into four call note categories, A, C, D, and Dh notes, based on their acoustic structure as observed in sound spectrograms, and evaluated based on the syntactical ordering of the note types within calls. The notes were then analyzed using quantitative measures and it was determined which features have the potential to convey information to discriminate note type, individual, and the geographic origin of the producer. The findings were comparable to previous research of congeners in that chestnut backed chickadee calls were produced with a relatively fixed syntax and contained similarly structured note types across all geographic regions. Overall this information will form a base for future research on chestnut-backed chickadee vocalizations and will strengthen the foundation for future comparative evolutionary studies. PMID- 19813819 TI - Sound pressure and particle acceleration audiograms in three marine fish species from the Adriatic Sea. AB - Fishes show great variability in hearing sensitivity, bandwidth, and the appropriate stimulus component for the inner ear (particle motion or pressure). Here, hearing sensitivities in three vocal marine species belonging to different families were described in terms of sound pressure and particle acceleration. In particular, hearing sensitivity to tone bursts of varying frequencies were measured in the red-mouthed goby Gobius cruentatus, the Mediterranean damselfish Chromis chromis, and the brown meagre Sciaena umbra using the non-invasive auditory evoked potential-recording technique. Hearing thresholds were measured in terms of sound pressure level and particle acceleration level in the three Cartesian directions using a newly developed miniature pressure-acceleration sensor. The brown meagre showed the broadest hearing range (up to 3000 Hz) and the best hearing sensitivity, both in terms of sound pressure and particle acceleration. The red-mouthed goby and the damselfish were less sensitive, with upper frequency limits of 700 and 600 Hz, respectively. The low auditory thresholds and the large hearing bandwidth of S. umbra indicate that sound pressure may play a role in S. umbra's hearing, even though pronounced connections between the swim bladder and the inner ears are lacking. PMID- 19813820 TI - Simulation of shear wave propagation in a soft medium using a pseudospectral time domain method. AB - Elastography applications require the use of efficient models to simulate the propagation of shear waves in soft media such as human tissues. These models are needed to improve understanding of the measured displacement field, to reconstruct the viscoelasticity of heterogeneous tissues, and to test inversion algorithms. This paper reports a numerical model based on a pseudospectral time domain method developed to simulate shear and compression wave propagation in an axisymmetric heterogeneous viscoelastic medium. This model was adapted to the study of soft tissues where the ratio between the compression and the shear wave velocity was about a thousand and validated in the homogeneous situation by comparison with an analytical model based on elastodynamic Green's functions. Displacements obtained experimentally using transient elastography are presented, compared with simulation results, and discussed. PMID- 19814536 TI - Molecular structure determination from x-ray scattering patterns of laser-aligned symmetric-top molecules. AB - We investigate the molecular structure information contained in the x-ray diffraction patterns of an ensemble of rigid CF(3)Br molecules aligned by an intense laser pulse at finite rotational temperature. The diffraction patterns are calculated at an x-ray photon energy of 20 keV to probe molecular structure at angstrom-scale resolution. We find that a structural reconstruction algorithm based on iterative phase retrieval fails to extract a reliable structure. However, the high atomic number of Br compared with C or F allows each diffraction pattern to be treated as a hologram. Using this approach, the azimuthal projection of the molecular electron density about the alignment axis may be retrieved. PMID- 19814537 TI - Unified expression for the rate constant of the bridged electron transfer derived by renormalization. AB - Electron transfer (ET) from a donor to an acceptor through an energetically close intermediary state on a midway molecule is a process found often in natural and artificial solar-energy capturing systems such as photosynthesis. This process has often been thought of in terms of opposing "superexchange" and "sequential or hopping" mechanisms, and the recent theory of Sumi and Kakitani (SK) [J. Phys. Chem. B 105, 9603 (2001)] has shown an interpolation between these mechanisms. In their theory, however, dynamics governing the most interesting intermediary region between them has artificially been introduced by phenomenologies. The dynamics is played by decoherence among electronic states, their decay, and thermalization of phonons in the medium. The present work clarifies the dynamics on a microscopic basis by means of renormalization in electronic coupling among the states, and gives a complete unified expression of the rate constant of the ET. It merges to that given by the SK theory in the semiclassical approximation for phonons interacting with an electron transferred. PMID- 19814538 TI - Generalized Langevin models of molecular dynamics simulations with applications to ion channels. AB - We present a new methodology, which combines molecular dynamics and stochastic dynamics, for modeling the permeation of ions across biological ion channels. Using molecular dynamics, a free energy profile is determined for the ion(s) in the channel, and the distribution of random and frictional forces is measured over discrete segments of the ion channel. The parameters thus determined are used in stochastic dynamics simulations based on the nonlinear generalized Langevin equation. We first provide the theoretical basis of this procedure, which we refer to as "distributional molecular dynamics," and detail the methods for estimating the parameters from molecular dynamics to be used in stochastic dynamics. We test the technique by applying it to study the dynamics of ion permeation across the gramicidin pore. Given the known difficulty in modeling the conduction of ions in gramicidin using classical molecular dynamics, there is a degree of uncertainty regarding the validity of the MD-derived potential of mean force (PMF) for gramicidin. Using our techniques and systematically changing the PMF, we are able to reverse engineer a modified PMF which gives a current-voltage curve closely matching experimental results. PMID- 19814539 TI - Modified adiabatic approximation: charge asymmetry in HD+ and HD. AB - The modified adiabatic approximation is discussed, in which the interaction of electrons with nuclei is partitioned between the electronic and nuclear Hamiltonian, in order to simulate the finite nuclear mass effect. The proposed formalism is universal and can be used in calculations for molecules of any size. The effect of electron localization on the deuteron in vibrationally excited states of HD(+) and the permanent dipole moment of HD, typically both explained in terms of nonadiabatic couplings between g and u states, are well reproduced with this method. PMID- 19814541 TI - Using the ONIOM hybrid method to apply equation of motion CCSD to larger systems: benchmarking and comparison with time-dependent density functional theory, configuration interaction singles, and time-dependent Hartree-Fock. AB - Equation of motion coupled-cluster singles and doubles (EOM-CCSD) is one of the most accurate computational methods for the description of one-electron vertical transitions. However, its O(N(6)) scaling, where N is the number of basis functions, often makes the study of molecules larger than 10-15 heavy atoms prohibitive. In this work we investigate how accurately less expensive methods can approximate the EOM-CCSD results. We focus on our own N-layer integrated molecular orbital molecular mechanics (ONIOM) hybrid scheme, where the system is partitioned into regions which are treated with different levels of theory. For our set of benchmark calculations, the comparison of conventional configuration interaction singles (CIS), time-dependent Hartree-Fock (TDHF), and time-dependent density functional theory (TDDFT) methods and ONIOM (with different low level methods) showed that the best accuracy-computational time combination is obtained with ONIOM(EOM:TDDFT), which has a rms of the error with respect to the conventional EOM-CCSD of 0.06 eV, compared with 0.47 eV of the conventional TDDFT. PMID- 19814540 TI - Exploration of effective potential landscapes using coarse reverse integration. AB - We describe a reverse integration approach for the exploration of low-dimensional effective potential landscapes. Coarse reverse integration initialized on a ring of coarse states enables efficient navigation on the landscape terrain: Escape from local effective potential wells, detection of saddle points, and identification of significant transition paths between wells. We consider several distinct ring evolution modes: Backward stepping in time, solution arc length, and effective potential. The performance of these approaches is illustrated for a deterministic problem where the energy landscape is known explicitly. Reverse ring integration is then applied to noisy problems where the ring integration routine serves as an outer wrapper around a forward-in-time inner simulator. Two versions of such inner simulators are considered: A Gillespie-type stochastic simulator and a molecular dynamics simulator. In these "equation-free" computational illustrations, estimation techniques are applied to the results of short bursts of inner simulation to obtain the unavailable (in closed-form) quantities (local drift and diffusion coefficient estimates) required for reverse ring integration; this naturally leads to approximations of the effective landscape. PMID- 19814542 TI - Comprehensive investigation about the second order term of thermodynamic perturbation expansion. AB - Monte Carlo simulations are carried out for the second order term in the thermodynamic perturbation expansion around a hard sphere reference fluid. The sample potentials considered cover a wide spectrum: From two frequently employed, namely hard sphere plus square well potential and hard core attractive Yukawa potential, to two kinds of repulsive potentials, namely hard sphere plus square shoulder potential and hard sphere plus triangle shoulder potential; the investigated potential range also extends from extremely short range to rather long range. The obtained simulation data are used to evaluate performance of two theoretical approaches, i.e., a traditional macroscopic compressibility approximation (MCA) and a recent coupling parameter expansion. Extensive comparison shows that the coupling parameter expansion provides a reliable method for accurately calculating the second order term of the high temperature series expansion, while the widely accepted MCA fails quantitatively or even qualitatively for most of the situations investigated. PMID- 19814543 TI - Inversion of radial distribution functions to pair forces by solving the Yvon Born-Green equation iteratively. AB - We develop a new method to invert the target profiles of radial distribution functions (RDFs) to the pair forces between particles. The target profiles of RDFs can be obtained from all-atom molecular dynamics (MD) simulations or experiments and the inverted pair forces can be used in molecular simulations at a coarse-grained (CG) scale. Our method is based on a variational principle that determines the mean forces between CG sites after integrating out the unwanted degrees of freedom. The solution of this variational principle has been shown to correspond to the Yvon-Born-Green (YBG) equation [Noid et al., J. Phys. Chem. B 111, 4116 (2007)]. To invert RDFs, we solve the YBG equation iteratively by running a CG MD simulation at each step of iteration. A novelty of the iterative YBG method is that during iteration, CG forces are updated according to the YBG equation without imposing any approximation as is required by other methods. As a result, only three to ten iterations are required to achieve convergence for all cases tested in this work. Furthermore, we show that not only are the target RDFs reproduced by the iterative solution; the profiles of the three-body correlation function in the YBG equation computed from all-atom and CG simulations also have a better agreement. The iterative-YBG method is applied to compute the CG forces of four molecular liquids to illustrate its efficiency and robustness: water, ethane, ethanol, and a water/methanol mixture. Using the resulting CG forces, all of the target RDFs observed in all-atom MD simulations are reproduced. We also show that the iterative-YBG method can be applied with a virial constraint to expand the representability of a CG force field. The iterative-YBG method thus provides a general and robust framework for computing CG forces from RDFs and could be systematically generalized to go beyond pairwise forces and to include higher-body interactions in a CG force field by applying the aforementioned variational principle to derive the corresponding YBG equation for iterative solution. PMID- 19814544 TI - Quantum mechanical scoring for protein docking. AB - We develop a docking protocol based on quantum mechanical/molecular mechanical calculations in which quantum mechanical energy is used as scoring. We test the protocol with three groups of examples with various binding site characteristics. The new docking method performs as well as or better than conventional docking methods in all three groups. In particular, for proteins with primarily hydrophobic binding sites, structural motifs with possible pi-pi interactions are often found and it is shown that these can be better modeled with quantum mechanical scoring docking than force field based methods. It seems that the new method performs in such cases to a great accuracy. PMID- 19814545 TI - Toward improved density functionals for the correlation energy. AB - Eleven density functionals, including some of the most widely used ones, are tested on their ability to predict nonrelativistic, electron correlation energies for the 17 atoms from He to Ar, the 17 cations from Li(+) to K(+), and 11 (1)S state atoms from Ca to Rn. They all lead to relatively poor predictions for the heavier atoms. Reparametrization of these functionals improves their performance for light atoms but does not alleviate their problems with the heavier, closed shell atoms. Several novel, few-parameter, density functionals for the correlation energy are developed heuristically. Four new functionals lead to qualitatively improved predictions for the heavier atoms without unreasonably compromising accuracy for the lighter atoms. Further progress would be facilitated by reliable estimates of electron correlation energies for more atoms, particularly heavy ones. PMID- 19814547 TI - Dissociative charge exchange dynamics of HN2(+) and DN2(+). AB - Translational spectroscopy coupled with coincidence detection techniques has been used to study the dissociation dynamics of ground state H/D+N(2) products resulting from charge exchange between keV beams of HN(2)(+)/DN(2)(+) and cesium. Analysis of the product kinetic energy release suggests that dissociation of HN(2) and DN(2) proceeds from initial populations in the (2)A("), 2 (2)A('), and 3s Rydberg electronic states of the neutral molecule. Although all three excited electronic states must eventually couple to the 1 (2)A(') ground state of HN(2)/DN(2), the resulting dissociation dynamics exhibit a significant dependence on the initial electronic state. Potential mechanisms are discussed in light of the observed product kinetic energy release distributions. PMID- 19814546 TI - Optimal estimators and asymptotic variances for nonequilibrium path-ensemble averages. AB - Existing optimal estimators of nonequilibrium path-ensemble averages are shown to fall within the framework of extended bridge sampling. Using this framework, we derive a general minimal-variance estimator that can combine nonequilibrium trajectory data sampled from multiple path-ensembles to estimate arbitrary functions of nonequilibrium expectations. The framework is also applied to obtain asymptotic variance estimates, which are a useful measure of statistical uncertainty. In particular, we develop asymptotic variance estimates pertaining to Jarzynski's equality for free energies and the Hummer-Szabo expressions for the potential of mean force, calculated from uni- or bidirectional path samples. These estimators are demonstrated on a model single-molecule pulling experiment. In these simulations, the asymptotic variance expression is found to accurately characterize the confidence intervals around estimators when the bias is small. Hence, the confidence intervals are inaccurately described for unidirectional estimates with large bias, but for this model it largely reflects the true error in a bidirectional estimator derived by Minh and Adib. PMID- 19814548 TI - A theoretical study of water equilibria: the cluster distribution versus temperature and pressure for (H2O)n, n = 1-60, and ice. AB - The size distribution of water clusters at equilibrium is studied using quantum chemical calculations in combination with statistical thermodynamics. The necessary energetic data is obtained by quantum-chemical B3LYP computations and through extrapolations from the B3LYP results for the larger clusters. Clusters with up to 60 molecules are included in the equilibrium computations. Populations of different cluster sizes are calculated using both an ideal gas model with noninteracting clusters and a model where a correction for the interaction energy is included analogous to the van der Waals law. In standard vapor the majority of the water molecules are monomers. For the ideal gas model at 1 atm large clusters [56-mer (0-120 K) and 28-mer (100-260 K)] dominate at low temperatures and separate to smaller clusters [21-22-mer (170-280 K) and 4-6-mer (270-320 K) and to monomers (300-350 K)] when the temperature is increased. At lower pressure the transition from clusters to monomers lies at lower temperatures and fewer cluster sizes are formed. The computed size distribution exhibits enhanced peaks for the clusters consisting of 21 and 28 water molecules; these sizes are for protonated water clusters often referred to as magic numbers. If cluster-cluster interactions are included in the model the transition from clusters to monomers is sharper (i.e., occurs over a smaller temperature interval) than when the ideal gas model is used. Clusters with 20-22 molecules dominate in the liquid region. When a large icelike cluster is included it will dominate for temperatures up to 325 K for the noninteracting clusters model. Thermodynamic properties (C(p), DeltaH) were calculated with in general good agreement with experimental values for the solid and gas phase. A formula for the number of H-bond topologies in a given cluster structure is derived. For the 20-mer it is shown that the number of topologies contributes to making the population of dodecahedron-shaped cluster larger than that of a lower-energy fused prism cluster at high temperatures. PMID- 19814549 TI - The photoelectron spectrum of the ethoxide anion: conical intersections, the spin orbit interaction, and sequence bands. AB - The negative ion photoelectron spectrum of the ethoxide anion (ethoxide-h(5)) and that of its fully deuterated analog, ethoxide-d(5), are calculated using the multimode vibronic coupling approach. A two state quasidiabatic Hamiltonian H(d) is constructed which includes all terms through second order in the full 18 dimensional internal coordinate space. H(d) is centered at the ab initio determined minimum energy crossing (MEX) point on the symmetry-allowed (2)A(") (2)A(') accidental seam of conical intersection and determined from ab initio energy gradients and derivative couplings. It reproduces the local topography of the (2)A(")-(2)A(') MEX, in addition to accurately representing the geometries, energetics, and harmonic frequencies of equilibrium and saddle point structures located on the ground electronic state potential energy surface in the vicinity of the MEX. Spin-orbit effects are included. The results for ethoxy-h(5) are compared to photoelectron and slow electron velocity-map imaging (SEVI) spectra. By comparing the measured and predicted photoelectron spectrum, the accuracy of the electronic structure treatment is inferred. The existence of sequence bands in the SEVI spectrum is established. PMID- 19814550 TI - Theoretical investigation of the potential energy surface of the van der Waals complex CH4-N2. AB - The interaction potential energy surface of the van der Waals CH(4)-N(2) complex has been calculated for a broad range of intermolecular separations and configurations in the approximation of rigid interacting molecules at the CCSD(T) and MP2 levels of theory using the correlation consistent aug-cc-pVTZ basis set. The BSSE correction was taken into account for all the calculations. The most stable configurations of the complex were found. Binding energies were calculated in the CBS limit with accounting for the molecular deformations. The harmonic and anharmonic fundamental vibrational frequencies and rotational constants for the ground and first excited vibrational states were calculated for the most stable configurations at the MP2 level of theory with BSSE correction. Fitting parameters were found for the most stable configuration for the Lennard-Jones and Esposti-Werner potentials. PMID- 19814551 TI - Homogeneous nucleation with magic numbers: aluminum. AB - Homogeneous nucleation of clusters that exhibit magic numbers is studied numerically, using as an example aluminum at 2000 K, based on recent calculations of free energies [Li et al., J. Phys. Chem. C 111, 16227 (2007)] and condensation rate constants [Li and Truhlar, J. Phys. Chem. C 112, 11109 (2008)] that provide a database for Al(i) up to i=60. The nucleation behavior for saturation ratios greater than about 4.5 is found to be dominated by a peak in the free energy change associated with the reaction iAl-->Al(i) at i=55, making it the critical size over a wide range of saturation ratios. Calculated steady-state nucleation rates are many orders of magnitude lower than predicted by classical nucleation theory (CNT). The onset of nucleation is predicted to occur at a saturation ratio of about 13.3, compared to about 5.1 in CNT, while for saturation ratios greater than about 25 the abundance of magic-numbered clusters becomes high enough to invalidate the assumption that cluster growth occurs solely by monomer addition. Transient nucleation is also predicted to be substantially different than predicted by CNT, with a much longer time required to reach steady state: about 10(-4) s at a saturation ratio of 20, compared to about 10(-7) s from CNT. Magic numbers are seen to play an important role in transient nucleation, as the nucleation currents for clusters of adjacent sizes become equal to each other in temporally successive groups, where the largest cluster in each group is the magic-numbered one. PMID- 19814552 TI - Multidimensional steric effects for the XeI* (B, C) formations in the oriented Xe* (3P(2),M(J) = 2) + oriented CF3I reaction. AB - Steric effects for the XeI(*) (B) and XeI(*) (C) formations in the oriented Xe(*) ((3)P(2),M(J)=2)+oriented CF(3)I reaction have been observed as a function of the mutual configuration between molecular orientation and atomic alignment in the collision frame. The mutual configuration exercises the significant influences on the stereoanisotropy for both the reactivity and the branching to the XeI(*) (B) and XeI(*) (C) channels. PMID- 19814553 TI - Photochemistry of the water dimer: time-dependent quantum wave-packet description of the dynamics at the S1-S0 conical intersection. AB - The photoinduced electron-driven proton-transfer dynamics of the water-dimer system has been investigated by time-dependent quantum wave-packet calculations. The main nuclear degrees of freedom driving the system from the Frank-Condon region to the S(0)-S(1) conical intersection are the distance between the oxygen atoms and the displacement of the hydrogen atom from the oxygen-oxygen bond center. Two important coupling modes have been investigated: Rotation of the H donating water dangling proton and asymmetric stretching of the H-accepting water dangling protons' O(a)H bonds. Potential energy surfaces of the ground and lowest excited electronic states have been constructed on the basis of ab initio calculations. The time-dependent quantum wave-packet propagation has been employed within the (2 + 1)-dimensional systems for the description of the nonadiabatic dynamics of water dimer. The effects of the initial vibrational state of the system on the electronic population transfer and dissociation dynamics are presented. To approximate the photochemical behavior of water dimer in bulk water, we add a boundary condition into the (2 + 1)-dimensional systems to simulate the existence of water bulk. The results provide insight into the mechanisms of excited state deactivation of the water-dimer system in gas phase and in bulk water through the electron-driven proton-transfer process. PMID- 19814555 TI - Penning ionization and ion fragmentation of formamide HCONH2 by He*, Ne*, and Ar* in molecular beams. AB - Mass spectra from Penning ionization by metastable atom bombardment (MAB) in the title system at kinetic energies near 1 kcal/mol are reported. The experiments employ a supersonic excited noble gas beam crossing an effusive beam of formamide vapor. Product ions are extracted perpendicular to the plane of the beams, analyzed by a quadrupole mass filter, and counted by a scintillation-type ion counter. Relative to 70 eV electron impact, the He(*) and Ne(*) spectra show more extensive breakage of C-N and C-H bonds despite the smaller available energy, while the Ar(*) spectrum shows only the molecular ion (m/z 45), H atom elimination (44), and the decarbonylation products CO+NH(3)(+) (17). Fragmentation in the latter system has been analyzed using a combination of ab initio calculations and Rice-Ramsperger-Kassel-Marcus theory with tunneling correction; good agreement with the experimental 45/44/17 intensity ratio 100/6.8+/-0.7/6.2+/-1.7 is obtained. 15% of m/z 17 and 50% of m/z 44 is attributed to tunneling. The ab initio decarbonylation reaction path yields a hydrogen bonded H(2)N-HCO(+) transition state, which transfers a proton while proceeding downhill to the observed products, while both the path and the energetics support the earlier conclusion that the lowest lying electronically excited state of the ion (2pi or 2a(")) crosses the ground state early along the reaction path, thereby dominating the dynamics of decarbonylation. PMID- 19814554 TI - Femtosecond pump-probe photoionization-photofragmentation spectroscopy: photoionization-induced twisting and coherent vibrational motion of azobenzene cation. AB - We report studies of ultrafast dynamics of azobenzene cation using femtosecond photoionization-photofragmentation spectroscopy. In our experiments, a femtosecond pump pulse first produces an ensemble of azobenzene cations via photoionization of the neutrals. A delayed probe pulse then brings the evolving ionic system to excited states that ultimately undergo ion fragmentation. The dynamics is followed by monitoring either the parent-ion depletion or fragment ion formation as a function of the pump-probe delay time. The observed transients for azobenzene cation are characterized by a constant ion depletion modulated by a rapidly damped oscillatory signal with a period of about 1 ps. Theoretical calculations suggest that the oscillation arises from a vibration motion along the twisting inversion coordinate involving displacements in CNNC and phenyl-ring torsions. The oscillation is damped rapidly with a time constant of about 1.2 ps, suggesting that energy dissipation from the active mode to bath modes takes place in this time scale. PMID- 19814556 TI - Ab initio quantum Monte Carlo study of the positronic hydrogen cyanide molecule. AB - Quantum Monte Carlo methods are used to investigate the binding of a positron to the hydrogen cyanide (HCN) and lithium hydride (LiH) molecules. Our value of the adiabatic positron affinity (PA) of LiH of 1.010(3) eV is very close to the best theoretical value of 1.005 eV, obtained from variational calculations using explicitly correlated Gaussian basis sets [K. Strasburger, J. Chem. Phys. 114, 00615 (2001)]. We have obtained a reliable estimate of 0.0378(48) eV for the PA of the HCN molecule, which is almost 20 times larger than that obtained at the Hartree-Fock level, and strongly supports the binding of a positron in the electrostatic field of the HCN molecule. Our results show the importance of correlation effects for describing weakly bound positronic molecular complexes. PMID- 19814557 TI - Imaging transient species in the femtosecond A-band photodissociation of CH3I. AB - A nonresonant femtosecond laser pulse centered at 802 nm is used to probe the real time photodissociation dynamics of CH(3)I in the A-band at 267 nm. Using multiphoton ionization with this probe laser pulse and velocity map ion imaging of CH(3)(+), we have followed the time evolution of the translational energy and spatial anisotropy of the CH(3) fragment, which in turn has permitted to image the C-I bond breaking from the initial Franck-Condon region up to the final products along the reaction coordinate. Given the temporal width of our pump and probe laser pulses (approximately 80 fs), a mechanism is proposed by which transient species are probed by simultaneous absorption of pump and probe laser pulses through intermediate Rydberg and ionic states of CH(3)I while the pump and probe pulses overlap in time. This study shows how the combination of femtosecond multiphoton ionization and ion imaging techniques provides an ideal tool to resolve in time the different stages of the bond breaking event in a polyatomic molecule. PMID- 19814558 TI - Linear and nonlinear optical properties of a series of Ni-dithiolene derivatives. AB - Some linear and nonlinear optical (NLO) properties of Ni(SCH)(4) and several of its derivatives have been computed by employing a series of basis sets and a hierarchy of methods (e.g., HF, DFT, coupled cluster, and multiconfigurational techniques). The electronic structure of Ni(SCH)(4) has been also analyzed by using CASSCF/CASPT2, ab initio valence bond, and DFT methods. In particular we discuss how the diradicaloid character (DC) of Ni(SCH)(4) significantly affects its NLO properties. The quasidegeneracy of the two lowest-energy singlet states 1 (1)A(g) and 1 (1)B(1u), the clear DC nature of the former, and the very large number of low-lying states enhance the NLO properties values. These particular features are used to interpret the NLO properties of Ni(SCH)(4). The DC of the considered derivatives has been estimated and correlated with the NLO properties. CASVB computations have shown that the structures with Ni(II) are the dominant ones, while those with Ni(0) and Ni(IV) have negligible weight. The weights of the four diradical structures were discussed in connection with the weight of the structures, where all the electrons are paired. Comparative discussion of the properties of Ni(SCH)(4) with those of tetrathia fulvalene demonstrates the very large effect of Ni on the properties of the Ni-dithiolene derivatives. A similar remarkable effect on the NLO properties is produced by one or two methyl or C(3)S groups. The considered Ni-dithiolene derivatives have exceptionally large NLO properties. This feature in connection with their other physical properties makes them ideal candidates for photonic applications. PMID- 19814559 TI - Structure, spectroscopic properties, and photochemistry of the hydroxymethoxy radical. AB - The hydroxymethoxy (HMO) radical is proposed to be the primary product of photodissociation of the atmospherically important hydroxymethyl hydroperoxide (HMHP). This transient species is still unknown and the present study provides theoretical predictions of properties, spectroscopy, and photochemistry of this radical for the first time. Structures, harmonic frequencies, vertical and vibrationally resolved absorption spectra are computed for several electronic states, using state-of-the-art ab initio electronic structure methods. The lowest excited state, absorbing in the mid to near infrared, seems to be the most promising candidate for spectroscopic identification of HMO. The electron affinity of 2.232 eV and the characteristic photodetachment spectrum is also predicted to be suitable for experimental investigations. By contrast, the B state absorbing around 3.5 eV is proposed to undergo fast photodissociation, forming CH(2)O and OH, and thus appears less useful for spectroscopic detection of HMO. However, the photodissociation may be important for the atmospheric chemistry of HMHP. Ionization of HMO will also lead to dissociation or rearrangement of the cation and will yield only unspecific spectra. PMID- 19814560 TI - Phase behavior of a de-ionized binary mixture of charged spheres in the presence of gravity. AB - We report on the phase behavior of an aqueous binary charged sphere suspension under exhaustively de-ionized conditions as a function of number fraction of small particles p and total number density n. The mixture of size ratio Gamma=0.557 displays a complex phase diagram. Formation of bcc crystals with no compositional order dominates. We observe a region of drastically decreased crystal stability at 0.55p>0.95 crystal formation is partially assisted by gravity, i.e., gravitational separation of the two species precedes crystal formation for samples in the coexistence range. In the composition range corresponding to the decreased crystal stability only lower bounds of the freezing and melting line are obtained, but the general shape of the phase diagram is retained. At p=0.93 and n=43 microm(-3) two different crystalline phases coexist in the bulk, while at p=0.4 additional Bragg peaks appear in the static light scattering experiments. This strongly suggests that we observe an eutectic in the region of decreased stability, while the enhanced stability at p=0.4 seems to correlate with compound formation. PMID- 19814561 TI - The velocity, refractive index, and equation of state of liquid ammonia at high temperatures and high pressures. AB - The high temperature and high pressure Brillouin scattering studies of liquid ammonia have been performed in a diamond anvil cell. Acoustic velocity, refractive index, adiabatic bulk modulus, and the equation of state of liquid ammonia were determined at temperatures up to 410 K and at pressures up to the solidification point. Velocity and refractive index increase smoothly with increasing pressure along isothermals but decrease slightly with the temperature increase. The bulk modulus increases linearly with pressure and its slope dB/dP decreases slightly with increasing temperature from 6.67 at 297 K to 5.94 at 410 K. PMID- 19814562 TI - Influence of temperature on coherent dynamics of a two-level system immersed in a dissipative spin bath. AB - The quantum dynamics of a two-level system coupled to an Ohmic spin bath is studied by means of the perturbation approach based on a unitary transformation. A scattering function xi(k) is introduced in the transformation to take into account quantum fluctuations. By the master equation within the Born approximation, nonequilibrium dynamics quantities are calculated. The method works well for the coupling constant 0 or =17 years) with BTcP. The study was conducted at 36 centers across the USA. The study comprised a 2-week open-label titration phase, followed by a double-blind efficacy phase, during which patients received sublingual fentanyl citrate orally disintegrating tablet (sublingual fentanyl ODT) or placebo, in a random order. The primary efficacy endpoint was the sum of pain intensity difference (SPID) over 30 min post-administration. Secondary efficacy endpoints included pain intensity difference (PID) and pain relief (PR) throughout the 60-min post-dose assessment period. Following efficacy evaluation, patients entered a long-term safety phase of up to 12 months. Adverse events were recorded throughout the study. [ CLINICAL TRIAL REGISTRATION: NCT00262678] RESULTS: A total of 131 patients entered the titration phase, of whom 61 were included in the primary efficacy analysis. Sublingual fentanyl ODT provided significant improvements in SPID relative to placebo at 30 min (49.5 vs. 36.6, p = 0.0004) and 60 min post-administration (143.0 vs. 104.5, p = 0.0002). Furthermore, sublingual fentanyl ODT provided significant improvements in PID and PR compared to placebo, from 10 min post-dose (p = 0.0055 and p = 0.049 for PID and PR, respectively). Patient recruitment was stopped early, due to positive interim analysis results (significant at prespecified level, p < or = 0.0414). Overall, sublingual fentanyl ODT was well-tolerated both systemically and sublingually, with 41 patients experiencing > or =1 study drug-related adverse event (AE). The most common AEs included nausea (12.2%), vomiting (5.3%) and somnolence (4.6%). One serious AE (mild affect lability) was considered possibly related to study medication. The observed pattern of AEs was consistent with that previously observed with fentanyl. CONCLUSIONS: Sublingual fentanyl ODT was efficacious and well-tolerated for the treatment of BTcP in opioid-tolerant patients with cancer. Sublingual fentanyl ODT provided significant improvements in pain intensity compared to placebo, from 10 min post-administration and throughout the 60-min post-dose assessment period. Sublingual fentanyl ODT was well tolerated over 12 months of treatment. PMID- 19814588 TI - rhBMP2/7 heterodimer: an osteoblastogenesis inducer of not higher potency but lower effective concentration compared with rhBMP2 and rhBMP7 homodimers. AB - Heterodimeric bone morphogenetic proteins (BMPs) were exhibited to be more potent than and thus potential substitutes for homodimeric BMPs whose clinical application is limited for the drawbacks resulted from their higher effective doses. This study aims to delineate the biofunctional characteristics of recombinant human BMP2/7 (rhBMP2/7) heterodimer in inducing osteoblastogenesis of MC3T3-E1 through in vitro time-course and dose-response studies. rhBMP2/7 heterodimer induced cell migration with a significantly lower optimal concentration and higher peak effect than the respective homodimers. rhBMP2/7 heterodimer induced cell differentiation with significantly lower threshold concentrations but similar maximum effects. On day 28, the area of calcium depositions induced by 50 ng/mL rhBMP2/7 was 12- or 38-fold more than that of 50 ng/mL rhBMP2 or 50 ng/mL rhBMP7, respectively. The results indicated that rhBMP2/7 heterodimer was an osteoblastogenesis inducer of not higher potency but lower effective concentration compared with rhBMP2 and rhBMP7 homodimers. PMID- 19814587 TI - Scleral reinforcement through host tissue integration with biomimetic enzymatically degradable semi-interpenetrating polymer network. AB - Enzymatically degradable semi-interpenetrating polymer networks (edsIPNs) were explored for their biocompatibility and ability to promote new scleral tissue growth, as a means of reinforcing the posterior wall of the eye. The edsIPNs comprised thermoresponsive poly(N-isopropylacrylamide-co-acrylic acid), customizable peptide crosslinkers cleavable by matrix metalloproteinases, and interpenetrating linear poly(acrylic acid)-graft-peptide chains to engage with cell surface receptors. Rheological studies revealed an increase in stiffness at body temperature; the complex shear modulus |G*| was 14.13 +/- 6.13 Pa at 22 degrees C and 63.18 +/- 12.24 Pa at 37 degrees C, compatible with injection at room temperature. Primary chick scleral fibroblasts and chondrocytes cultured on edsIPN increased by 15.1- and 11.1-fold, respectively, over 11 days; both exhibited delayed onset of exponential growth compared with the cells plated on tissue culture polystyrene. The edsIPN was delivered by retrobulbar injection (100 microL) to nine 2-week-old chicks to assess biocompatibility in vivo. Ocular axial dimensions were assessed using A-scan ultrasonography over 28 days, after which eyes were processed for histological analysis. Although edsIPN injections did not affect the rate of ocular elongation, the outer fibrous sclera showed significant thickening. The demonstration that injectable biomimetic edsIPNs stimulate scleral fibrous tissue growth represents proof-of-principle for a novel approach for scleral reinforcement and a potential therapy for high myopia. PMID- 19814589 TI - Age-related changes in material behavior of porcine mitral and aortic valves and correlation to matrix composition. AB - Recent studies showing significant changes in valvular matrix composition with age offer design criteria for age-specific tissue-engineered heart valves. However, knowledge regarding aging-related changes in valvular material properties is limited. Therefore, 6-week, 6-month, and 6-year-old porcine aortic valves (AV) and mitral valves (MV) were subjected to uniaxial tensile testing. In addition to standard material parameters, the radius of transition curvature (RTC) was measured to assess the acuteness of the transition region of the tension-strain curve. Radially, the MV had greater stiffness and a smaller RTC compared with the AV. Circumferentially, the center of the MV anterior leaflet (MVAC) had the highest stiffness (MVAC > AV > MV free edge [MVF]), greater stress relaxation (MVAC > MVF/AV), lowest extensibility (MVAC < AV < MVF), and smaller RTC compared with MVF (AV < MVAC < MVF). AV and MV radial strips had a larger RTC compared with circumferential strips. Aging elevated stiffness for MV and AV radial and circumferential strips, elevated stress relaxation in AV and MVF circumferential strips, and increased RTC for MV radial and MVF circumferential strips. In conclusion, there are significant age-related differences in the material properties of heart valves, which parallel differences in tissue composition and structure, likely impact valve function, and highlight the need for age-specific design goals for tissue-engineered heart valves. PMID- 19814590 TI - Localization of the potential zonal marker clusterin in native cartilage and in tissue-engineered constructs. AB - An increasing number of studies aim to engineer cartilage tissue that more closely mimic the zonal organization of native articular tissue. Further understanding of zonal differences is crucial for successful development and evaluation of such grafts. We therefore aimed to characterize the secretion of the potential zonal marker clusterin by zonal articular chondrocytes in osteoarthritic and healthy articular cartilage and in tissue-engineered constructs. Clusterin secreted by superficial, middle, and deep zones equine chondrocytes was immunolocalized in cytospins of alginate cultured superficial, middle, and deep zones equine chondrocytes. Clusterin was present within the superficial zone of native cartilage; after isolation of the cells from healthy articular cartilage, staining for clusterin was limited to cells derived from the superficial zone. Staining disappeared after expansion, but reappeared during (re)differentiation and was more pronounced within the cultures derived from the superficial zones of the cartilage. The presence of clusterin was associated with clusters of differentiating chondrocytes, rather than highly proliferative cells and did not specifically colocalize with proteoglycan-4. Because staining for clusterin was more abundant in cultures of superficial chondrocytes compared to those of cells of the deeper layers, it may be used to further characterize zonal cartilage constructs. PMID- 19814591 TI - The nitric oxide donor S-nitrosoglutathione reduces apoptotic primary liver cell loss in a three-dimensional perfusion bioreactor culture model developed for liver support. AB - INTRODUCTION: Artificial extracorporeal support for hepatic failure has met with limited clinical success. In hepatocytes, nitric oxide (NO) functions as an antiapoptotic modulator in response to a variety of stresses. We hypothesized that NO administration would yield improved viability and hepatocellular restructuring in a four-compartment, hollow fiber-based bioreactor with integral oxygenation for dynamic three-dimensional perfusion of hepatic cells in bioartificial liver support systems. METHODS: Isolated adult rat liver cells were placed in culture medium alone (control) or medium supplemented with various concentrations of an NO donor (S-nitrosoglutathione [GSNO]) in the bioreactors. Media samples were obtained from the cell perfusion circuit to monitor cellular response. After 24 and 72 h, histology biopsies were taken to investigate spontaneous restructuring of the cells. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to quantify apoptotic nuclei. RESULTS: Control bioreactors exhibited 47.9 +/- 2.9% (mean +/- standard error of the mean) apoptotic nuclei. In contrast, NO-treated bioreactors exhibited a biphasic response. Fewer apoptotic nuclei were seen in the 200 and 500 microM GSNO groups (14.4 +/- 0.4%). No effect was observed in the 10 microM GSNO group (47.3%), and increased TUNEL staining was observed in the 1000 microM GSNO group (82.6%). Media lactate dehydrogenase levels were lower in bioreactor groups treated with 200 or 500 microM GSNO (310 +/- 38 IU/L) compared with the control group (919 +/- 188 IU/L; p < 0.05). Protein synthesis was not affected, as measured by albumin levels in the media (115 +/- 19 microg/day/cell inoculum in GSNO-treated bioreactors at 24 h vs. 110 +/- 13 in controls; p = 0.851). Histologically, all of the bioreactor groups exhibited liver cell aggregates with some attached to the bioreactor capillaries. Increased numbers of cells in the aggregates and superior spontaneous restructuring of the cells were seen at 24 and 72 h in the bioreactor groups treated with either 200 or 500 microM GSNO compared with the control groups. CONCLUSION: Addition of an NO donor reduces adult rat liver cell apoptosis during the initial 24 h after cell inoculation within a three-dimensional perfusion bioreactor system for liver support and promotes liver cell aggregation and spontaneous restructuring of the cells at 24 and 72 h. GSNO-treated bioreactors remain metabolically active and show significantly lower levels of cellular injury as compared with controls. Further studies will be required to evaluate the impact of NO treatment of liver support bioreactors for clinical studies. PMID- 19814592 TI - Synthesis and antimicrobial studies of s-triazine based heterocycles. AB - In an effort to discover new candidates with improved antimicrobial activities we report here the synthesis and in vitro biological evalution of various series of 2-(N-methylamino)-4-(N,N-dimethylamino)-6-(arylthioureido)-s-triazine (5a-j) and (N-methylamino)-4-(N,N-dimethylamino)-6-(arylureido)-s-triazine (6a-j). All the synthesized compounds were screened in vitro for their antibacterial activity against two different gram-positive bacteria (S. aureus, B. subtilis) and two different gram-negative bacteria (P. aeruginosa, E. coli) using the broth dilution method. PMID- 19814593 TI - Cervical cancer screening among Asian Canadian immigrant and nonimmigrant women. AB - OBJECTIVES: To examine whether Pap smear screening and determinants of screening for Asian immigrants are different from those for nonimmigrants in Canada. METHODS: Data from the Canadian Community Health Survey Cycle 2.1 (2003) were used. Explanatory and multivariate logistic regression analyses were performed to compare rates and determinants of Pap smear screening between Asian immigrants and nonimmigrants. RESULTS: In this study, 64,604 women were included. Asian immigrants had significantly lower rates of Pap smear screening (52%) compared to nonimmigrants (72%). Lack of necessity and time are important barriers. CONCLUSION: Pap smear screening should be promoted, particularly in recent Asian immigrants. PMID- 19814594 TI - BOUNCE: an exploratory healthy lifestyle summer intervention for girls. AB - OBJECTIVES: To assess the efficacy of the BOUNCE parent-daughter intervention in promoting selected physical fitness measures and activity. METHODS: Thirty-seven Latino and African American parent-daughter pairs participated. The intervention entailed physical activities, nutrition classes, and self-esteem activities. Child's height, weight, waist circumference, body fat %, blood pressure, physical fitness, and activity were assessed pre- and postintervention. RESULTS: A repeated-measures ANOVA yielded significant results [F(4,26) = 793.003, P < 0.001]. Post hoc analyses demonstrated significant (P < 0.05) changes in BMI, waist circumference, physical fitness, and activity. CONCLUSIONS: The BOUNCE intervention was effective in promoting fitness and activity in minority girls. PMID- 19814595 TI - Testing social cognitive mechanisms of exercise in college students. AB - OBJECTIVES: To determine if different dimensions of outcome expectations are mediators between self-efficacy and exercise among college students. METHODS: Students (n = 290) reported exercise levels, self-efficacy, and expectations using a Web-based questionnaire. Factor analysis was performed to identify dimensions of expectations; regression analyses tested whether each factor mediated the relationship between self-efficacy and exercise. RESULTS: Physical, mental, social, and self-evaluative expectations emerged from the factor analysis, but none was associated with exercise independent of self-efficacy. Self-efficacy, however, was significantly associated with exercise independent of expectations (P < 0.01). CONCLUSIONS: College students' activity expectations are multidimensional, but do not mediate the relationship between self-efficacy and exercise. PMID- 19814596 TI - Assessment of pregnancy cigarette smoking and factors that predict denial. AB - OBJECTIVES: To determine rates of pregnancy smoking concealment compared to behavioral observation and to identify factors predicting untruthful denial. METHODS: Review of 843 delivery charts. RESULTS: Based on observation during delivery hospitalization, 8% of smokers denied hospital admission, 16% denied throughout prenatal care. Compared with those admitting smoking, false deniers had higher levels of education, incomes and adequate prenatal care utilization; and were less likely to have drug use, STDs, or hepatitis C. Observation was a valid tool for assessing smoking status. CONCLUSIONS: Typical concealers were those considered lower risk, increasing the chance they would go undetected as smokers. PMID- 19814597 TI - Use of program evaluation in community youth tobacco cessation programs. AB - OBJECTIVE: To examine the inclusion of program evaluation components in a national sample of youth tobacco cessation programs. METHODS: Program administrators were interviewed to ascertain program characteristics associated with the inclusion of program evaluation components. RESULTS: Two thirds of all surveyed programs (n=591) had an evaluation component; most included attendance, user satisfaction, and quitting measures. Programs with an evaluation component were significantly more likely to report annual funding greater than the median and to conduct a follow-up with participants. CONCLUSION: Program characteristics and associated evaluation components are summarized, and recommendations are given for a minimal level of program evaluation planning. PMID- 19814599 TI - Health risk perception, optimistic bias, and personal satisfaction. AB - OBJECTIVES: To examine change in risk perception and optimistic bias concerning behavior-linked health threats and environmental health threats between adolescence and young adulthood and how these factors related to personal satisfaction. METHODS: In 1996 and 2002, 1624 adolescents responded to a mailed questionnaire. RESULTS: Adolescents showed strong positive optimistic bias concerning behaviorlinked risks, and this optimistic bias increased with age. Increase in optimistic bias over time predicted increase in personal satisfaction. CONCLUSIONS: The capacity to process and perceive potential threats in a positive manner might be a valuable human ability positively influencing personal satisfaction and well-being. PMID- 19814598 TI - Individual and job-related variation in infant feeding practices among working mothers. AB - OBJECTIVE: To document working mothers' infant feeding practices and delineate factors that may shape infant feeding. METHODS: Cross-sectional data were obtained from a community sample of working women with 8-month old infants (n=199). RESULTS: Nearly all working mothers used commercially prepared foods like infant cereals, fruits, and vegetables. Approximately one-fifth fed infants french fries, sweetened beverages, and sweetened desserts. Unhealthy infant feeding was elevated among unmarried mothers, those with less education, and those with a nonstandard work schedule. CONCLUSIONS: Working mothers use commercially prepared foods for infant feeding. Socially disadvantaged working mothers' infant feeding may pose health and developmental risks. PMID- 19814600 TI - Key principles in internet-based weight management systems. AB - OBJECTIVE: To provide a review of weight management components as a means for developing suggested guidelines for online program development or selection. METHODS: Forty-two published studies associated with Internet programs were reviewed. RESULTS: Studies included online weight management components: tailored feedback, social support, self-monitoring, and interactive components. Five key principles associated with Internet-based weight management programming are presented: (1) recreate the human experience, (2) personalize to the individual, (3) create a dynamic experience, (4) provide a supportive environment, and (5) build upon sound theory. CONCLUSIONS: Key principles can guide in the development or selection of online weight management programs. PMID- 19814601 TI - Identity consolidation and health risk behaviors in college students. AB - OBJECTIVES: To investigate the protective role of personal identity consolidation against health risk behaviors in college-attending emerging adults. METHODS: A multisite sample of 1546 college students completed measures of personal identity consolidation and recent risk behavior engagement. RESULTS: Multivariate Poisson regression indicated that personal identity consolidation was negatively related to binge drinking, illicit drug use, sexual risk behaviors, and risky driving. These findings were consistent across gender, ethnicity, and place of residence. CONCLUSIONS: A consolidated sense of personal identity may protect college attending emerging adults from health-compromising behaviors. Health professionals could incorporate an identity development component into college health programming. PMID- 19814602 TI - Understanding physical activity maintenance in breast cancer survivors. AB - OBJECTIVES: To examine predictors of physical activity (PA) 6 months after a randomized controlled trial. METHODS: Breast cancer survivors (N=377) completed assessments at baseline, postintervention, and 6 months after the intervention. Data were collected on demographic, medical, psychosocial, motivational, and behavioral variables at baseline, postintervention, and 6 months. RESULTS: Survivors meeting PA guidelines at baseline (OR = 5.54, 95% CI = 2.99-9.82, P < 0.001) and postintervention (OR = 2.92, 95% CI = 1.63-5.23, P < 0.001) had a greater likelihood of meeting PA guidelines at 6 months follow-up. CONCLUSIONS: Interventions promoting PA maintenance should focus on optimizing PA behavior, psychosocial functioning, and motivation. PMID- 19814604 TI - Factors related to adolescent drinking in Appalachia. AB - OBJECTIVES: To examine the relationships among parental monitoring, perceptions of peer drinking, and adolescent alcohol consumption. METHODS: Tenth- and 12th grade students (N=648) in a rural, Appalachian county were surveyed. RESULTS: A binomial logistic regression revealed a composite of those who had perceptions that many peers drank, low parental monitoring, and no biological male guardian in the home were 8.496 times more likely to have ever been drunk. Other characteristics resulted in lower odds. CONCLUSIONS: Parental monitoring and perceptions of peer drinking were important predictors of drinking in this rural sample. Prevention efforts in school and at home should address both variables. PMID- 19814603 TI - Adolescent sleep, risk behaviors, and depressive symptoms: are they linked? AB - OBJECTIVE: To explore how weekday and weekend sleep patterns are related to adolescent substance use, depressive symptoms, and school truancy. METHODS: Selfreport surveys of 242 youth (93.4% white, mean age 16.4 years). RESULTS: Longer weekday sleep duration was inversely associated with depressive symptoms, past month alcohol use, and drunkenness. Later weekend bedtime and wake-times, compared to those of weekdays, were associated with increased substance use and truancy. CONCLUSIONS: Weekday sleep duration appears to be protective for substance use, depression and school truancy for teenagers. However, inconsistent sleep patterns between weekdays and weekends were associated with a range of markers for adolescent risk. PMID- 19814605 TI - Twenty five years of poison center experience with propoxyphene overdoses the FDA could not find. PMID- 19814606 TI - Renal failure after ingestion of Amanita proxima. PMID- 19814607 TI - Development of a physiologically based kinetic model for 99m-technetium-labelled carbon nanoparticles inhaled by humans. AB - Particulate air pollution is associated with respiratory and cardiovascular morbidity and mortality. Recent studies investigated whether and to which extent inhaled ultrafine particles are able to translocate into the bloodstream in humans. However, their conclusions were conflicting. We developed a physiologically based kinetic model for (99m)technetium-labelled carbon nanoparticles (Technegas). The model was designed to analyse imaging data. It includes different translocation rates and kinetics for free technetium, and small and large technetium-labelled particles. It was calibrated with data from an experiment designed to assess the fate of nanoparticles in humans after inhalation of Technegas. The data provided time courses of radioactivity in the liver, stomach, urine, and blood. Parameter estimation was performed in a Bayesian context with Markov chain Monte Carlo (MCMC) techniques. Our analysis points to a likely translocation of particle-bound technetium from lung to blood, at a rate about twofold lower than the transfer rate of free technetium. Notably, restricting the model so that only free technetium would have been able to reach blood circulation resulted in much poorer fits to the experimental data. The percentage of small particles able to translocate was estimated at 12.7% of total particles. The percentage of unbound technetium was estimated at 6.7% of total technetium. To our knowledge, our model is the first PBPK model able to use imaging data to describe the absorption and distribution of nanoparticles. We believe that our modeling approach using Bayesian and MCMC techniques provides a reasonable description on which to base further model refinement. PMID- 19814608 TI - Rapidly growing mycobacteria: emerging pathogens in cosmetic procedures of the skin. PMID- 19814609 TI - Clinical management of rapidly growing mycobacterial cutaneous infections in patients after mesotherapy. AB - BACKGROUND: Increasing numbers of patients are expressing an interest in mesotherapy as a method of reducing body fat. Cutaneous infections due to rapidly growing mycobacteria are a common complication of such procedures. METHODS: We followed up patients who had developed cutaneous infections after undergoing mesotherapy during the period October 2006-January 2007. RESULTS: Sixteen patients were infected after mesotherapy injections performed by the same physician. All patients presented with painful, erythematous, draining subcutaneous nodules at the injection sites. All patients were treated with surgical drainage. Microbiological examination was performed on specimens that were obtained before and during the surgical procedure. Direct examination of skin smears demonstrated acid-fast bacilli in 25% of the specimens that were obtained before the procedure and 37% of the specimens obtained during the procedure; culture results were positive in 75% of the patients. Mycobacterium chelonae was identified in 11 patients, and Mycobacterium frederiksbergense was identified in 2 patients. Fourteen patients were treated with antibiotics, 6 received triple therapy as first-line treatment (tigecycline, tobramycin, and clarithromycin), and 8 received dual therapy (clarithromycin and ciprofloxacin). The mean duration of treatment was 14 weeks (range, 1-24 weeks). All of the patients except 1 were fully recovered 2 years after the onset of infection, with the mean time to healing estimated at 6.2 months (range, 1-15 months). CONCLUSIONS: This series of rapidly growing mycobacterial cutaneous infections highlights the difficulties in treating such infections and suggests that in vitro susceptibility to antibiotics does not accurately predict their clinical efficacy. PMID- 19814610 TI - Cruise ship environmental hygiene and the risk of norovirus infection outbreaks: an objective assessment of 56 vessels over 3 years. AB - BACKGROUND: Norovirus infection outbreaks (NoVOs) occur frequently in closed populations, such as cruise ship passengers. Environmental contamination is believed to play an important role in NoVO propagation. METHODS: Trained health care professionals covertly evaluated the thoroughness of disinfection cleaning (TDC) of 6 standardized objects (toilet seat, flush handle or button, toilet stall inner handhold, stall inner door handle, restroom inner door handle, and baby changing table surfaces) with high potential for fecal contamination in cruise ship public restrooms, by means of a previously validated novel targeting method. RESULTS: Fifty-six cruise ships (approximately 30% of 180 vessels operated by 9 large cruise lines) were evaluated from July 2005 through August 2008. Overall, 37% (range, 4%-100%; 95% confidence interval, 29.2%-45.4%) of 8344 objects in 273 randomly selected public restrooms were cleaned daily. The TDC did not differ by cruise line and did not correlate with the Centers for Disease Control and Prevention (CDC) Vessel Sanitation Program inspection scores (r(2), .002; P = .75). More than half the vessels had overall TDC scores <30%, although several of these low-scoring ships had near-perfect CDC sanitation scores. The mean TDC of the 3 ships evaluated within 4 months before a NoVO (10.3%) was substantially less than the mean TDC of the 40 ships that did not experience NoVOs (40.4%) (P < .004). CONCLUSIONS: An objective evaluation of public restroom environmental hygiene on 56 cruise ships found that only 37% of selected toilet area objects were cleaned on a daily basis. Low TDC scores may predict subsequent NoVO-prone vessels. Enhanced public restroom cleaning may prevent or moderate NoVOs on cruise ships. PMID- 19814612 TI - Collaboration: the foundation for success in rare disease genetic test development. PMID- 19814613 TI - New trends in molecular biomarker discovery for breast cancer. AB - Breast cancer is one of the most common and leading causes of cancer death in women. Early diagnosis, selection of appropriate therapeutic strategies, and efficient follow-up play an important role in reducing mortality. Recently, HER 2/neu in breast cancer has been routinely used to guide treatment of using Trastuzumab in less than 25-30% of patients. More new biomarkers will be still expected in the future to tailor treatments. However, there are still many obstacles in developing clinically useful biomarker tests for clinical practice. A lack of specificity of tumor markers and lack of sensitivity of testing systems have been noticed, which limit their clinical use. Finding biomarkers for breast cancer could allow physicians to identify individuals who are susceptible to certain types and stages of cancer to tailor preventive and therapeutic modalities based on the genotype and phenotype information. These biomarkers should be cancer specific, and sensitively detectable in a wide range of specimen(s) containing cancer-derived materials, including body fluids (plasma, serum, urine, saliva, etc.), tissues, and cell lines. This review highlights the new trends and approaches in breast cancer biomarker discovery, which could be potentially used for early diagnosis, development of new therapeutic approaches, and follow-up of patients. PMID- 19814614 TI - Design and validation of a conformation-sensitive capillary electrophoresis system for mutation identification of the COL7A1 gene with automated peak comparison. AB - Dystrophic epidermolysis bullosa is a heritable skin disease in which blisters occur because of a defect in type VII collagen resulting from mutations in the COL7A1 gene that is composed of 118 exons. Although a few mutations are specific to certain populations owing to founder effects, and although a few mutational hotspots exist, most mutations are unique to families and can be found scattered throughout the entire COL7A1 gene. This emphasizes the need for a sensitive, reliable, and efficient mutation scanning technique. Therefore, we developed a conformation-sensitive capillary electrophoresis (CSCE) system for COL7A1 mutation scanning. Here we report on the design and validation of this system. The CSCE technique is based on the principle of heteroduplex formation when polymerase chain reaction-amplified DNA fragments containing heterozygous sequence changes are slowly reannealed. These fluorescently labeled fragments have different migration characteristics and can be detected on a multi-capillary automated sequencer. Validation was performed by analysis of 29 known COL7A1 sequence changes, covering 33% of amplicons. After optimization of the conditions, all 29 sequence changes were detected by the CSCE system, irrespective of length or CG-content of amplicons and position of sequence changes, reflecting an analytical sensitivity of 90.2-100% (95% confidence interval). We conclude that this CSCE system is a rapid, reliable, cost effective, and highly sensitive way of mutation scanning for COL7A1 in a molecular genetics service laboratory. PMID- 19814615 TI - Comparison of the cytogenetic and molecular analyses in the assessment of imatinib response in chronic myelocytic leukemia. AB - We aimed to compare the cytogenetic and molecular analyses in the assessment of imatinib mesylate response in patients suffering the chronic phase of chronic myelocytic leukemia who were refractory to alpha-interferon treatment. A total of 117 patients in the chronic phase of chronic myelocytic leukemia were included. The patients were treated with 400 mg/day imatinib mesylate. Bone marrow samples were obtained for the cytogenetic and molecular analyses. Patients without the Ph chromosome were defined as complete cytogenetic responders. Partial cytogenetic response was determined when the Ph chromosome was detected in 1-35% of the cells. Molecular response was determined by quantitative real-time reverse transcriptase polymerase chain reaction (QR-PCR) and defined as no detection of BCR-ABL mRNA. The frequencies of complete and partial cytogenetic response were 29% (n = 34) and 15% (n = 18), respectively. No cytogenetic response was achieved in 56% (n = 65) of the patients. Molecular response was achieved in 62% (n = 21) and 33% (n = 6) of the complete and partial cytogenetic responders, respectively. All of the 65 patients with no cytogenetic response were also molecular nonresponders. We conclude that there is reasonable agreement between the cytogenetic and molecular analyses. Both methods are complementary in the assessment of response to therapy. PMID- 19814616 TI - Possible association of combined vitamin D receptor and estrogen receptor genotypes and low bone mineral density in Jordanian postmenopausal women. AB - The genetics of osteoporosis has been extensively studied over the last 20 years. Many of the studies have been aimed at identifying possible risk factors and possible association with low bone mineral density (BMD). Vitamin D receptor (VDR) and estrogen receptor (ER) gene polymorphisms were the first to be studied. Some studies have shown a possible association for individual VDR and ER or combined VDR and ER genotypes in some populations, and others showed lack of such an association. This study is aiming at identification of a possible association with low BMD in Jordanian postmenopausal women. We used restriction fragment length polymorphisms (RFLPs) to study four polymorphisms in the VDR gene and two polymorphisms in the ER gene. Our sample was composed of 100 normal controls and 120 samples from patients with symptomatic vertebral fractures. The results showed a possible association of the ppxx genotype with low BMD in controls and patients and an association of the AaBbTT genotype with high BMD in control subjects. The AABBTT, AABBTTFF, and AABBTTFFPPXX genotypes showed a possible association with low BMD in patients. Further studies are needed to confirm the last finding since it could be an important predictor of low BMD in the Jordanian population. PMID- 19814617 TI - Diagnosis of Russell-Silver syndrome by the combined bisulfite restriction analysis-denaturing high-performance liquid chromatography assay. AB - Russell-Silver syndrome (RSS) is characterized by prenatal and postnatal growth retardation, triangular facies, and fifth-finger clinodactyly. Half of all patients with RSS have hypomethylation of the differentially methylated region of the H19 gene on chromosome 11p15.5. Hence, a quantitative methylation analysis of this region can be useful for the molecular diagnosis of RSS. However, conventional assays based on bisulfite clone sequencing are rather time and labor consuming and are not suitable for clinical use. In the present study, we investigated a possible method of quantitatively determining H19 hypomethylation in RSS patients using a combined bisulfite restriction analysis (COBRA) denaturing high-performance liquid chromatography (DHPLC) assay; in this combined assay, polymerase chain reaction products amplified from the H19 differentially methylated region of bisulfite-treated genomic DNA were analyzed using a COBRA assay, which detects methylation-dependent sequence differences in the bisulfite treated genomic DNA using a restriction enzyme analysis. We designed the assay so that a restriction enzyme (HinfI) would cut the methylated, but not the unmethylated, template. The molar ratio between the cut and uncut fragments was measured using DHPLC, and the construction of a calibration curve enabled the methylation index for the original genomic DNA to be estimated. An analysis of seven RSS patients using the COBRA-DHPLC assay demonstrated that three of the seven RSS patients had a low methylation index of around 10%. A comparison of the methylation indices obtained using COBRA-DHPLC and conventional bisulfite clone sequencing revealed an excellent intermethod agreement. In summary, we have developed a robust, rapid, and cost-effective COBRA-DHPLC-based screening system for RSS. PMID- 19814618 TI - The analysis of the relationship between A1298C and C677T polymorphisms of the MTHFR gene with prostate cancer in Eskisehir population. AB - Prostate cancer is the most common cause of cancer deaths in men and is a major health problem worldwide. Methylene tetrahydrofolate reductase (MTHFR) plays an important role for folate metabolism and is also an important source for DNA methylation and DNA synthesis (nucleotide synthesis). The objective of this study was to investigate the relationship between the A1298C and C677T polymorphisms of the MTHFR gene and prostate cancer in the Turkish population. In our study, 93 prostate cancer patients between the ages of 50-89 and a control group of 166 benign prostate hyperplasia patients were evaluated. C677T and A1298C polymorphism ratios were compared among these two groups, and an analysis was made to see if there is a statistically meaningful difference. In this study, it has been observed that C677T polymorphism of the MTHFR gene produces no statistically significant difference for T allele frequency and the genotype frequency in prostate cancer patients and male controls with benign prostate hyperplasia not having prostate cancer, whereas it has been observed that A1298C polymorphism produces a statistically significant difference for C allele frequency in prostate cancer patients and controls and that it also produces a statistically marginal significance for genotype frequencies. PMID- 19814619 TI - The association of MMP-9 enzyme activity, MMP-9 C1562T polymorphism, and MMP-2 and -9 and TIMP-1, -2, -3, and -4 gene expression in lung cancer. AB - AIM: To investigate the association of gene expression of MMP-2 and -9, and TIMP 1, -2, -3, and -4 and polymorphism frequencies of MMP-9 C1562T and plasma MMP-9 enzyme activity in lung cancer patients. METHODS: In this study, DNA and RNA samples were extracted from peripheral blood of 300 subjects (200 lung cancer patients and 100 controls). MMP-9 C1562T polymorphism was determined using restriction fragment length polymorphism (RFLP) method, and expression of MMP-2 and -9, TIMP-1, -2, -3, and -4 was determined using reverse transcriptase polymerase chain reaction. Plasma MMP-9 enzyme activity levels were measured using enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION: The frequencies of C1562T genotypes were found to be CC 67%, CT 30%, and TT 3% in the control group and CC 75%, CT 24%, and TT 1% in the patient group. It was determined that CC genotype frequency increases significantly in patients according to control group. Plasma MMP-9 enzyme activity levels increased in patients with lung cancer compared to the control group. The cut-off value of MMP-9 enzyme activity was determined as 7.76 ng/mL by receiver operating characteristics curve analysis. The sensitivity, specificity, positive predictive value, and negative predictive value were 77%, 51%, 75.9%, and 52.6%, respectively. The expression of MMP-2 and TIMP-1 was found to be higher in lung cancer patients. Finally, we claim that determination of MMP-9 enzyme levels and expression of MMP-2 and -9 and TIMP-1 can be used as a marker in lung cancer. PMID- 19814620 TI - GJB2 mutations in patients with nonsyndromic hearing loss from Croatia. AB - The aim of the study was to determine (1) the frequency and type of mutations in the coding region of the GJB2 gene (sequencing), (2) the frequency of splice site mutation IVS1 + 1G > A in the GJB2 gene (multiplex ligation-dependent probe amplification analysis), (3) possible copy number changes in the GJB2, GJB3, GJB6, and WFS1 genes (multiplex ligation-dependent probe amplification analysis), and (4) the frequency of del(GJB6-D13S1830) in the GJB6 gene in 58 unrelated patients with nonsyndromic hearing loss from Croatia. About 44.8% of our patients presented with mutation in the GJB2 gene. We identified seven sequence variations. Six of them had previously been reported as disease related (35delG, W24X, V37I, L90P, 313del14, and IVS1 + 1G > A), and we report here for the first time one novel variant, -24A > C. We detected the greatest frequency of 35delG allele compared to the other alleles (35.3%). Allelic frequencies of other common mutations accounted for 2.6-0.9% of analyzed chromosomes. Neither GJB6 deletion nor copy number changes in the GJB2, GJB3, GJB6, and WFS1 genes were found. The 35delG/35delG genotype was associated with severe to profound hearing loss in 94% of 35delG homozygotes. High mutation rate (44%) indicates that testing of the GJB2 gene will clarify the genetic cause in almost half of the cases of recessive nonsyndromic hearing loss in Croatia. PMID- 19814621 TI - Rate and timing of hepatitis C virus relapse after a successful course of pegylated interferon plus ribavirin in HIV-infected and HIV-uninfected patients. AB - Information on the rate and timing of hepatitis C virus (HCV) relapse after treatment with pegylated interferon plus ribavirin is scarce. Among 604 patients treated for chronic hepatitis C, the 386 who were human immunodeficiency virus (HIV) positive attained an end-of-treatment response less frequently and experienced relapse more often than did the 218 who were HIV negative. However, episodes of HCV relapse occurred before week 12 in most cases, regardless of HIV status. PMID- 19814622 TI - Pneumococcal conjugate vaccine, polysaccharide vaccine, or both for adults? We're not there yet. PMID- 19814623 TI - The R753Q polymorphism abrogates toll-like receptor 2 signaling in response to human cytomegalovirus. AB - Toll-like receptor 2 (TLR2) serves as a pattern recognition receptor that signals the presence of cytomegalovirus. Herein, we report that R753Q polymorphism paralyzes TLR2-mediated immune signaling in cells exposed to cytomegalovirus glycoprotein B. This immunologic impairment could serve as a biologic mechanism underlying the association between the TLR2 R753Q polymorphism and cytomegalovirus disease in humans. PMID- 19814624 TI - The immunogenicity of 7-valent pneumococcal conjugate vaccine versus 23-valent polysaccharide vaccine in adults aged 50-80 years. AB - BACKGROUND: Infections with pneumococci are a major cause of morbidity and mortality in the elderly population. Although 23-valent pneumococcal polysaccharide vaccine (PPV) is recommended for elderly persons, the potential benefits of conjugate vaccine use in this age group remain unclear. METHODS: We performed an open-label, randomized study that compared 7-valent pneumococcal conjugate vaccine (7vPnC) with PPV in 599 adults aged 50-80 years. Vaccinees received either 1 dose of 7vPnC or PPV or 1 dose of 7vPnC followed by a dose of 7vPnC or PPV 6 months later. Groups were stratified so they contained similar numbers of individuals aged 50-59, 60-69, and 70-80 years. Concentrations of immunoglobulin G specific for the serotypes in 7vPnC were measured before and 4-6 weeks after each vaccination and 1 year after enrollment. RESULTS: Although baseline antibody levels were slightly lower in the older age groups, responses (fold rises) to either vaccine did not depend on age. Single-dose 7vPnC was superior for only 3 serotypes. Administration of a second dose of PPV or 7vPnC was similarly immunogenic in adults primed with 7vPnC, and titers after a second dose were similar to the first. CONCLUSIONS: Pneumococcal vaccines retain their immunogenicity when administered into the eighth decade of life, but a second dose, when assessed by antibody titers alone, has little utility. 7vPnC vaccines do not lead to subsequent hyporesponsiveness. CLINICAL TRIALS REGISTRATION: http://www.clinicaltrials.gov/NCT00197821. PMID- 19814625 TI - Immunological and biochemical correlates of adjunctive dexamethasone in Vietnamese adults with bacterial meningitis. AB - Adjunctive treatment to improve outcome from bacterial meningitis has centered on dexamethasone. Among Vietnamese patients with bacterial meningitis, cerebrospinal fluid (CSF) opening pressure and CSF:plasma glucose ratios were significantly improved and levels of CSF cytokines interleukin (IL)-6, IL-8, and IL-10 and were all statistically significantly lower after treatment in patients who were randomized to dexamethasone, compared with levels in patients who received placebo. PMID- 19814626 TI - Very late-onset group B Streptococcus meningitis, sepsis, and systemic shigellosis due to interleukin-1 receptor-associated kinase-4 deficiency. AB - We describe a child with very late-onset group B Streptococcus sepsis and meningitis, systemic shigellosis, and chronic osteomyelitis. Peripheral blood cells obtained from the patient and her brother did not respond to stimulation with either interleukin-1beta or lipopolysaccharide. Sequencing of the interleukin-1 receptor-associated kinase-4 gene revealed 2 novel mutations. PMID- 19814628 TI - Circadian pattern and the effect of standardized physical exercise on procollagen IIA N-peptide (PIIANP) in rheumatoid arthritis at different stages and in healthy individuals. AB - BACKGROUND: Variant collagen IIA is re-expressed in diseased cartilage. Low procollagen IIA N-peptide (PIIANP) levels in serum have recently been reported in rheumatoid arthritis (RA). We investigated circadian rhythmicity and effect of physical activity on PIIANP in early and longstanding RA and in healthy subjects. METHODS: Patients with early and longstanding RA and controls were included. Fasting and serial blood samples were collected during 24 h. PIIANP response to physical activity was studied before and serially after standardized exercise. RESULTS AND CONCLUSION: In RA at different stages and healthy individuals, PIIANP exhibited no circadian rhythmicity, and PIIANP in serum was not influenced by physical activity. PMID- 19814627 TI - High rate of virologic suppression with raltegravir plus etravirine and darunavir/ritonavir among treatment-experienced patients infected with multidrug resistant HIV: results of the ANRS 139 TRIO trial. AB - BACKGROUND: The introduction of 2 or 3 fully active drugs in human immunodeficiency virus (HIV)-infected patients receiving failing antiretroviral therapy is a key determinant of subsequent treatment efficacy. The aim of this study was to assess the safety and efficacy of a regimen containing raltegravir, etravirine, and darunavir/ritonavir for treatment-experienced patients infected with multidrug-resistant HIV. METHODS: Patients enrolled in this phase II, noncomparative, multicenter trial were naive to the investigational drugs and had plasma HIV RNA levels >1000 copies/mL, a history of virologic failure while receiving nonnucleoside reverse-transcriptase inhibitors (NNRTI), > or =3 primary protease inhibitor and nucleoside reverse transcriptase inhibitor (NRTI) mutations, and < or =3 darunavir and NNRTI mutations. The primary end point was the proportion of patients with plasma HIV RNA levels <50 copies/mL at 24 weeks. RESULTS: A total of 103 patients enrolled in the study. At baseline, genotypic resistance profiles showed a median of 4 primary protease inhibitor mutations, 1 NNRTI mutation, and 6 NRTI mutations. In addition to the investigational drugs, 90 patients (87%) received optimized background therapy that included NRTIs (86 patients) or enfuvirtide (12 patients). At week 24, 90% of patients (95% confidence interval, 85%-96%) had an HIV RNA level <50 copies/mL. At week 48, 86% (95% confidence interval, 80%-93%) had an HIV RNA level <50 copies/mL. The median CD4 cell count increase was 108 cells/mm(3). Grade 3 or 4 clinical adverse events were reported in 15 patients (14.6%). Only 1 patient discontinued the investigational antiretroviral regimen, because of an adverse event. CONCLUSION: In patients infected with multidrug-resistant virus who have few remaining treatment options, the combination of raltegravir, etravirine, and darunavir/ritonavir is well tolerated and is associated with a rate of virologic suppression similar to that expected in treatment-naive patients. PMID- 19814629 TI - Trends in cardiac surgery-associated acute renal failure in the United States: a disproportionate increase after heart transplantation. AB - Acute renal failure (ARF) is common after cardiac surgery and more frequent after complex cardiac surgery. While the incidence of ARF is increasing after coronary artery bypass graft (CABG) surgery, trends in other forms of cardiac surgery remain unclear. We investigated the trend of ARF in various cardiac procedures and compared patterns using CABG surgery as a reference group. The study population consisted of discharges from the Nationwide Inpatient Sample from 1988 to 2003, grouped according to surgery as: CABG, CABG with mitral valve, CABG with other valve, valve alone, and heart transplant. Standard diagnostic codes were used to identify ARF among discharges. Multivariable regression was used to determine trends in ARF among various procedures with CABG as a reference group. The incidence of ARF increased in all five groups (p < 0.001) over the 16-year period. The ARF incidence was highest in the heart transplant group (17%). Compared to the CABG population, patients following heart transplantation developed ARF at higher rates during the study period. In contrast, while ARF increased over time in other groups, the rates of rise were slower than in CABG patients. Among heart surgery procedures, ARF incidence is highest in heart transplantation. The incidence of ARF is also increasing at a faster rate in this group of patients in contrast to other procedure groups when compared to CABG surgery. The disproportionate increase in ARF burden after heart transplantation is a concern due to its strong association with chronic kidney disease and mortality. PMID- 19814630 TI - The effect of strict volume control on cognitive functions in chronic hemodialysis patients. AB - Cognitive dysfunction is a well-known complication of chronic renal failure that is evident in 30% of hemodialysis (HD) patients. However, the pathogenesis of this dysfunction is unknown. Left ventricular hypertrophy could develop in hypertensive HD patients without establishing normovolemia. Our aim was to evaluate the effect of strict volume control by salt restriction and ultrafiltration on cognitive functions in HD patients. This cross-sectional study was composed of 22 HD patients who were normotensive by applying a strict volume control, 24 HD patients who were normotensive by receiving anti-hypertensive drugs, and 20 healthy controls. The strict volume control was defined as managing of blood pressure control by strict salt restriction and insistent ultrafiltration. P300 recording as an indicator of cognitive disfunction was measured when blood pressures were reached at target level at the end of six month follow-up period. In all patients, dimensions of the heart were evaluated with echocardiography on an interdialytic day. The cardiothoracic ratio and echocardiographic dimensions were significantly lower in patients with strict volume control. P300 amplitudes were significantly lower in patients on antihypertensive drugs than in patients with strict volume control (9.5 +/- 5.1 versus 11.3 +/- 5.4 muV). P300 latency was longer in patients on antihypertensive drugs than in the control group and patients with strict volume control (359.9 +/ 39.6 versus 345.6 +/- 36.7 ms). Our results suggest that hypervolemia may be one of the causal and potentially modifiable factors of cognitive dysfunction. Strict volume control may have beneficial effects on cognitive functions in hemodialysis patients. PMID- 19814631 TI - Trends in dialysis modality for individuals with acute kidney injury. AB - BACKGROUNDS: There are no national level data on types of dialysis in use for acute kidney injury (AKI). We aimed to assess trends in dialysis modality for AKI and mortality associated with each modality from 1998 to 2005. METHODS: Using data from the 5% Medicare cohort, we identified individuals with AKI requiring dialysis. Individuals with preexisting end-stage renal disease were excluded. Intermittent hemodialysis (IHD), daily intermittent hemodialysis, and continuous renal replacement therapy (CRRT) were defined using Current Procedure Terminology codes. Mortality was defined as death during 30 days after the first dialysis session. RESULTS: Between 1998 and 2005, there were a total of 18,249 patients identified with AKI requiring renal replacement therapy. CRRT was increasingly used for AKI, with 9.9% of patients in 1998 to 18.3% by 2005. Proportion of daily dialysis decreased during this period, while use of IHD remained stable at approximately 68%. Overall 30-day mortality declined from 44.4% in 1998 to 40.2% in 2005. Crude mortality for CRRT was highest in all years (51.0-61.8%), followed by daily (38.2-49.9%) and IHD groups (35.8-43.4%). Multinomial logistic regression analysis showed that white race, presence of sepsis, atherosclerotic heart diseases, peripheral vascular diseases, dysrhythmia, gastrointestinal and liver diseases, and any year after 2000 were independently associated with higher odds of using CRRT after adjusting for other variables. CONCLUSION: The proportion of patients using CRRT has increased over time. Mortality associated with IHD has decreased from 1998 to 2005. Mortality associated with different dialysis modalities is likely the result of severity of illness. PMID- 19814632 TI - The effect of the type of membrane on intradialytic complications and mortality in crush syndrome. AB - AIMS: There are not enough data about the type of the membrane that should be used in acute intermittent hemodialysis (IHD) in patients with crush syndrome where intradialytic complication rate is high. The effects of dialyzers on outcome have been investigated in this study. METHODS: Patients who required IHD due to crush syndrome after a big earthquake that struck Marmara in 1999 have been studied. Hemodynamic and biochemical analyses at the time of admission were examined. The patients were divided into three groups according to the type of dialyzers (viz., hemophan, polysulfone, and combined). RESULTS: Forty-five patients were included in the study (mean age: 33.9 +/- 13.3 years, mean HD session per patient: 8.8 +/- 6.1). In all, 408 dialyzers were used during IHD therapy (21% hemophan). The types of dialyzers used were hemophan (8 patients), polysulfone (18 patients), and the combination of the two (19 patients). The demographic and biochemical parameters related to crush syndrome were not different statistically. All sessions were anticoagulant-free. Hypotension and coagulation of sets were the main intradialytic complications. Five (11%) patients died, but there was no correlation between mortality rates and the type of the dialyzer used. Serum albumin, blood pressure, and thrombocyte counts were found to be related to mortality. CONCLUSION: No effect of the type of dialysis membrane on outcome was detected in patients with crush syndrome. Other potential factors, which may responsible for the complications and mortality, should be investigated. PMID- 19814633 TI - Superior outcome of nafamostat mesilate as an anticoagulant in patients undergoing maintenance hemodialysis with intracerebral hemorrhage. AB - AIMS: The incidence of complications associated with cerebrovascular diseases in patients who receive hemodialysis for a long-term period is higher than that of other complications. It is known that mortality due to cerebral hemorrhage is two times higher compared to non-dialysis patients. Anti-coagulants used for hemodialysis are essential. Accordingly, in cases in which the cerebral hemorrhage occurred, the selection of anti-coagulants for the prevention of further bleeding poses a great challenge to physicians. The change of hematoma and patient prognosis has a direct relationship. Many ongoing studies are conducted to examine the causative factors causing the increased hematoma and their related prognostic factors. In the current study, we examined the effect of nafamostat mesylate (a serine protease inhibitor) on the change of hematoma compared to heparin in hemodialysis patients. METHODS: The current study was conducted in 17 hemodialysis patients who developed a cerebral hemorrhage. These patients were assigned to two groups based on the type of anti-coagulants that they used (i.e., nafamostat mesylate and heparin). Then, the factors affecting the change of hematoma following the onset of cerebral hemorrhage were examined. The prognosis of hematoma was assessed based on brain CT scans, which were performed two weeks after the onset of cerebral hemorrhage in four groups. Following this, groups 1 (the decreased hematoma) and 2 (the decreased delay) were merged to group A (resolving group), and groups 3 (the increased hematoma) and 4 (the death following the aggravation) were merged to group B (the expansion group) for further analysis. RESULTS: There were no significant differences in baseline characteristics between the nafamostat group and the heparin group. A comparison between the resolving group and the expansion group also showed that there were no significant differences in baseline characteristics. In the anti coagulants and the change of hematoma, however, there were significant differences between the two groups (p = 0.024). A comparison of the change of hematoma between the four groups was also made. This showed that platelet counts and BUN level were significant factors (Platelet; p = 0.042, BUN; p = 0.043 ANOVA with resolving group). CONCLUSIONS: Nafamostat mesylate has a similar profile of anti-coagulative activity to heparin. It is assumed, however, that nafamostat has an affirmative effect on the recovery of damaged sites following the onset of cerebral hemorrhage. It is an anti-coagulant that can be safely used for hemodialysis following the onset of cerebral hemorrhage. PMID- 19814634 TI - The spectrum of adult postinfectious glomerulonephritis in the new millennium. AB - BACKGROUND: Postinfectious glomerulonephritis is rare in adults. The characteristics of the disease now differ from what were described decades ago. The goal of this study is to illustrate the clinicopathological spectrum of the disease in the modern era. METHODS: Between July 2000 and June 2008, 20 adult cases of postinfectious glomerulonephritis were identified at a medical center in Taiwan. The patients' records were retrospectively reviewed with respect to clinical presentation, microbiology, serology, morphology of renal biopsy, and clinical course. RESULTS: There were 14 males and 6 females. The mean age was 61 years. All patients developed acute renal failure, and the majority (65%) required dialysis support during the disease course. Hypocomplementemia was present in 60% of patients. The most frequently identified infectious agent was Staphylococcus (60%). Histological characteristics showed two distinct patterns of glomerulonephritis: diffuse endocapillary proliferative glomerulonephritis (65%) and focal mesangial proliferative glomerulonephritis (35%). There were no significant differences in the clinical presentation and outcome between the two groups. However, glomerular neutrophil infiltration was more commonly present in diffuse endocapillary proliferative pattern (p = 0.017). The percentage of patients with focal mesangial proliferative pattern significantly increased over time (p < 0.001). At the last follow-up, 6 patients (30%) had died, 6 (30%) were in complete remission, 4 (20%) had partial remission with renal insufficiency, and 4 (20%) were on chronic dialysis. CONCLUSIONS: Our data suggested that Staphylococcus had become the leading pathogen in adult postinfectious glomerulonephritis over the past 10 years. Furthermore, atypical histological feature with focal mesangial proliferative pattern was increasingly identified over time. The prognosis was still guarded, carrying a considerable mortality rate and risk for developing chronic renal failure. PMID- 19814635 TI - The effect of heavy muscle activity on renal cytoresistance in rats. AB - Cytoresistance is the term used to describe the response of the proximal tubule cells to various stress inducers via cholesterol accumulation. However, the role of extensive exercise as a renal insult has not been examined. In this study, the effect of heavy muscle activity on proximal tubule cytoresistance was investigated. Results obtained from rats subjected to running a treadmill for five days were compared to those of controls. Extensive muscle activity-induced soleus citrate synthase and blood lactate elevation were associated with normal MAP, RBF, and GFR. Blood electrolytes and cholesterol levels remained unchanged, whereas the total and free cholesterol accumulations in the proximal tubule cells of the exercised group were higher than controls. Cholesterol-loaded tubules were more resistant (as proved by LDH release) to an ATP-depleted/calcium overloaded second stress. These data clearly demonstrate that heavy muscle activity induces cholesterol accumulation in the proximal tubules of kidney, without influencing ATP generation. PMID- 19814636 TI - Protective effects of pycnogenol against ischemia reperfusion-induced oxidative renal injury in rats. AB - INTRODUCTION: Oxygen free radicals are involved in pathophysiology of ischemia/reperfusion (I/R) injury. This study was designed to assess the possible protective effect of pycnogenol (PYC) against I/R-induced oxidative renal damage. MATERIALS AND METHODS: Wistar albino rats were unilaterally nephrectomized and subjected to 45 min of renal pedicle occlusion followed by 3 h of reperfusion. PYC (10 mg kg(-1), i.p.) or saline was administered at 15 min prior to ischemia and immediately before the reperfusion period. At the end of the 3 h, rats were decapitated and trunk blood was collected. Creatinine, blood urea nitrogen (BUN), and lactate dehydrogenase (LDH) activity were measured in the serum samples, while proinflammatory cytokines, TNF-alpha, IL-1beta, and IL-6 levels were assayed in plasma samples. Kidney samples were taken for the determination of tissue malondialdehyde (MDA), glutathione (GSH) levels, Na+, K+-ATPase, and myeloperoxidase (MPO) activities, and the extent of tissue injury was analyzed microscopically. RESULTS: Ischemia/reperfusion caused a significant decrease in tissue GSH level and Na+, K+-ATPase activity, which was accompanied with significant increases in the renal MDA level and MPO activity. Similarly, serum creatinine and BUN levels, as well as LDH and IL-1beta, IL-6, and TNF-alpha levels, were elevated in the saline-treated I/R group as compared to saline treated control group. On the other hand, PYC treatment reversed all these biochemical indices, as well as histopathological alterations that were induced by I/R. CONCLUSIONS: Findings of the present study suggest that pycnogenol exerts renoprotective effects, via its free radical scavenging and antioxidant activities, that appear to involve the inhibition of tissue neutrophil infiltration. PMID- 19814637 TI - Effects of everolimus on cytokines, oxidative stress, and renal histology in ischemia-reperfusion injury of the kidney. AB - BACKGROUND: To evaluate the effects of everolimus on renal ischemia-reperfusion injury (IRI). METHODS: Wistar albino rats were divided into control, ischemia reperfusion (IR), and ischemia-reperfusion/everolimus (IR/eve) groups. Everolimus was administered for seven consecutive days to the IR/eve group prior to injury. IR and IR/eve groups underwent forty-five minutes ischemia followed by the application of reperfusion at 2 and 24 hours. Blood samples and kidneys were taken from all animals. RESULTS: . Serum blood urea nitrogen and creatinine levels increased at two hours of reperfusion in the IR and IR/eve groups, and decreased at 24 hours of reperfusion in the IR group. In the IR/eve group, we detected significantly high interleukin-6 levels and low tumor necrosis factor alpha and malondialdehyde levels at 24 hours. Myeloperoxidase levels increased at two hours of reperfusion in the IR/eve group, but decreased significantly at 24 hours. Everolimus did not improve renal tubular and interstitial injuries in renal IRI. CONCLUSIONS: It has been demonstrated that pretreatment with everolimus has beneficial effects on cytokines and oxidative stress in renal IRI. However, these effects are insufficient for the correction of histopathological changes and restoration of normal kidney function. PMID- 19814638 TI - Comparison of the efficacy of melatonin and 1400W on renal ischemia/reperfusion injury: a role for inhibiting iNOS. AB - INTRODUCTION: We investigated the roles of melatonin (a powerful antioxidant, iNOS inhibitor, and a scavenger of peroxynitrite) and 1400W (a strong and selective inhibitor of inducible nitric oxide) on renal dysfunction and injury induced by ischemia/reperfusion (I/R) of rat kidney, since oxidative and nitrosative injury are believed to be the major causes. MATERIALS AND METHODS: Thirty-two male Sprague-Dawley rats were divided into four groups of sham operated, I/R, I/R + Melatonin and I/R + 1400W. Rats were given either melatonin (10 mg/kg) or 1400W (10 mg/kg) in the I/R + Melatonin and I/R + 1400W groups respectively at 6 h prior to ischemia and at the beginning of reperfusion via intraperitoneal route. I/R injury was induced by 60 min of bilateral renal ischemia followed by 6 h of reperfusion. After reperfusion, kidneys and blood were obtained for histopathologic and biochemical evaluation. RESULTS: Melatonin and 1400W had an ameliorative effect on both oxidative and nitrosative stress in the kidneys against renal I/R injury in rats. In addition, melatonin significantly reduced elevated nitro-oxidative stress product, restored decreased antioxidant enzymes and attenuated histological alterations when compared with 1400W. CONCLUSIONS: Both Melatonin and 1400W were efficient in ameliorating experimental I/R injury of the kidneys. Moreover, melatonin was more effective than 1400W possibly through inhibiting iNOS as well as scavenging free oxygen radicals and peroxynitrite. PMID- 19814639 TI - Estrogen protects against oxidative multiorgan damage in rats with chronic renal failure. AB - The impact of sex dimorphism on chronic renal failure (CRF)-induced oxidative multiorgan damage and the effects of estradiol (E(2)) loss and E(2) supplementation on the progress of CRF were studied. Sprague-Dawley rats underwent 5/6 nephrectomy (CRF), and a group of female rats had bilateral ovariectomy (OVX), while the sham-operated rats had no nephrectomy or OVX. Rats received either estradiol propionate (50 microg/kg/day) or vehicle for six weeks. Serum BUN levels were elevated in both male and female CRF groups treated with vehicle, while creatinine level was not significantly changed in the female CRF group. CRF-induced elevation in serum TNF-alpha of male rats was abolished when the animals were treated with E(2), while OVX exaggerated TNF-alpha response. In OVX and male rats with CRF, E(2) treatment reversed the malondialdehyde elevations in all the studied tissues (kidney, heart, lung, ileum, brain, liver, and gastrocnemius muscle), while depletion of glutathione in these tissues was prevented by E(2) treatment. Similarly, increased levels of myeloperoxidase activity, lucigenin chemiluminescence, and collagen in most of the tissues were reversed by E(2) treatment. The findings show that the extent of tissue injuries was relatively less in females, while ovariectomy exacerbated all the indices of oxidative injury. Moreover, the administration of E(2), with its potent anti oxidant and anti-inflammatory effects, markedly improved CRF-induced systemic inflammatory outcomes in both male and female rats by depressing tissue neutrophil infiltration and modulating the release of inflammatory cytokines. PMID- 19814640 TI - The role of Na+-H+ exchanger isoform 1 in aldosterone-induced glomerulosclerosis in vivo. AB - Aldosterone is reported to promote fibrosis of multiple organs. Recent studies showed that Na+-H+ exchanger isoform 1 (NHE1) was involved in mineralocorticoid induced tissue fibrosis. The present study examined the role of NHE1 in aldosterone-induced glomerulosclerosis in rats. SD male rats were subjected to 5/6 nephrectomy and divided into four groups: rats subjected to sham operation were used as control (SHAM group), 5/6 nephrectomy (SNX group), SNX treated with aldosterone via osmotic mini-pump (ALDO group), and SNX treated with aldosterone plus NHE1 inhibitor 5-(N, N-Dimethyl) amiloride hydrochloride (DMA) (ALDO+DMA group). The rats were sacrificed at the 12th week. We found that aldosterone treatment significantly increased kidney weight/body weight ratio and systolic blood pressure compared with SNX rats. Aldosterone also increased proteinuria and serum creatinine level. The NHE1 antagonist DMA significantly reversed the effect of aldosterone on proteinuria, but had no effect on the aldosterone associated hypertension and the elevation of serum creatinine. The remnant kidney of 5/6 nephrectomized rats exhibited increased glomerulosclerosis score, tubulointerstitial fibrosis, and tubular proteinaceous cast, which were significantly enhanced by aldosterone treatment. DMA treatment significantly reduced aldosterone-associated glomerulosclerosis, but failed to improve aldosterone-induced tubulointerstitial fibrosis and tubular proteinaceous cast. The aldosterone-induced increase in renal TGFbeta1 and PCNA was significantly prevented by treatment with DMA. Our data showed that NHE1 inhibitor reduced aldosterone-induced glomerulosclerosis but not hypertension in 5/6 nephrectomized rats. The present study suggested that NHE1 contributed to aldosterone-induced glomerulosclerosis and could be a potential therapeutic target for chronic kidney disease. PMID- 19814641 TI - Acute tubular necrosis associated with non-hemorrhagic Dengue fever: a case report. AB - Dengue fever (DF) is an arthropod-born viral infection affecting humans. Dengue viruses are transmitted through the bites of the mosquito Aedes aegypti. Acute renal failure (ARF) is reported in patients who are affected mainly with Dengue hemorrhagic fever (DHF), which is a severe presentation of the disease. We report the case of a 24-year-old Omani female with no past history of particular medical problems. She was referred to our hospital for the further management of acute renal failure. She had clinical features of DF without DHF. The kidney biopsy showed features of acute tubular necrosis (ATN). She had a complete recovery after 25 days and required three sessions of hemodialysis. We conclude that DF even without DHF may lead to ATN and ARF. Clinicians should be aware of this etiology. Treatment is supportive and may require dialysis. The prognosis could be favorable. PMID- 19814642 TI - Preserved residual kidney function after twelve years' hemodialysis. AB - Residual kidney function (RKF) contributes significantly to solute clearance and fluid removal for dialysis patients, and the presence of RKF is associated with less morbidity and better long-term outcome. Most studies demonstrate that peritoneal dialysis preserves RKF better than hemodialysis (HD). Herein, we report a 55-year-old man with end stage renal failure who had been on chronic HD for 12 years. His RKF is preserved with very slow decline during the past years. Without specific intervention, delicate fluid management, minimal ultrafiltration, and stable hemodynamics during HD may help maintain his RKF. He is currently normotensive with good nutritional status. Although unexpected, we report this HD patient can preserve his RKF for at least 12 years. PMID- 19814643 TI - A case of progressive hypertension preceding gemcitabine-associated thrombotic microangiopathy complicated by acute kidney injury and stroke. AB - Gemcitabine-associated thrombotic microangiopathy is being increasingly recognized as a serious complication of treatment. We report a normotensive patient who developed progressive hypertension after commencing gemcitabine therapy. She also developed subtle changes in her platelet count and serum creatinine months before her emergent presentation. Clinicians should be aware of new onset or worsening hypertension and 'mild' biochemical changes in gemcitabine treated patients. PMID- 19814644 TI - Hydralazine-induced ANCA-positive pauci-immune glomerulonephritis: a case report and literature review. AB - We report a case of hydralazine-induced alveolar hemorrhage and anti-neutrophil cytoplasmic antibody (ANCA)-positive pauci-immune glomerulonephritis, with serum anti-histone antibodies present, features not previously described in the literature with this drug. A 50-year-old Caucasian female had hypertension treated with hydralazine 75mg TID for three years, and a lung nodule followed up periodically with chest-computed tomographies. She was admitted to the hospital for hemoptysis and newly discovered diffuse pulmonary ground-glass opacities. Transbronchial lung biopsy showed alveolar hemorrhage. Serum creatinine was 3.5 mg/dL and urinalysis showed 2+blood, 30-50RBC/hpf and red blood cell casts. ANCA against myeloperoxidase were present. Anti-double-stranded DNA, ANA, and anti histone antibodies were positive. Serum complements were normal. Renal biopsy revealed focal crescentic necrotizing glomerulonephritis with negative immunofluorescence, consistent with pauci-immune ANCA-positive vasculitis. Serum creatinine returned to baseline three days after hydralazine was discontinued, and the hemoptysis resolved after treatment with cyclophosphamide and prednisone was started. We concluded that this case represents a hydralazine-induced small vessel vasculitis rather than an idiopathic one. The possibility of hydralazine induced vasculitis should be considered when patients treated with hydralazine develop a pulmonary-renal syndrome. Anti-histone antibodies may be present in the absence of full classification criteria of drug-induced lupus. PMID- 19814645 TI - Acute kidney injury in patients with inactive cytochrome P450 polymorphisms. AB - Medications are a major source of acute kidney injury, especially in critically ill patients. Medication-induced renal injury can occur through a number of mechanisms. We present two cases of acute kidney injury (AKI) where inactive cytochrome P450 (CYP) polymorphism may have played a role. The first patient developed a biopsy-proven allergic interstitial nephritis following urethrotomy. Genetic testing revealed the patient to be heterozygous for an inactivating polymorphism CYP2C9*3 and homozygous for an inactivating polymorphism CYP2D6*4. Patient had received several doses of promethazine, which is metabolized by CYP2D6*4. Another patient developed AKI on several occasions after exposure to lansoprazole and allopurinol. CYP testing revealed the patient to be homozygous for inactivating polymorphism CYP2C19*2, which is responsible for the metabolism of lansoprazole. These are the first two cases of AKI associated with non functional polymorphisms of cytochrome P450 superfamily. While the exact mechanism has not been worked out, it introduced the possibility of a new source of kidney injury. PMID- 19814646 TI - Angio-embolization of renal artery pseudoaneurysm after renal biopsy: a case report. AB - Renal artery pseudoaneurysm is a rare clinical entity that has been reported after renal biopsy, percutaneous renal surgery, penetrating trauma, and rarely blunt renal trauma. We present the case of a 37-year-old man with ruptured renal artery pseudoaneurysm accompanied by massive gross hematuria, urinary clot retention, and bladder tamponade, which were the presenting signs seven hours after renal biopsy. Abdominal CT scan showed a large perinephric, intracapsular hematoma of left kidney. His angiogram revealed a left renal segmental artery pseudoaneurysm that measured 1 cm x 1 cm. He was successfully treated by selective embolization of the arterial branch supplying the pseudoaneurysm. PMID- 19814647 TI - Acute renal failure associated with acute non-fulminant hepatitis A: a case report and review of literature. AB - Most of the cases of non-fulminant hepatitis A carry good renal and overall prognosis. Here we present a case of acute renal failure associated with acute non-fulminant hepatitis A, for which kidney biopsy showed interstitial nephritis and tubular necrosis. We also review the literature and possible pathogenesis. PMID- 19814648 TI - Challenges of genomics and proteomics in nephrology. AB - An increasing number of patients suffering from renal diseases and limitations in standard diagnostic and therapeutic approaches has created an intense interest in applying genomics and proteomics in the field of nephrology. Genomics has provided a vast amount of information, linking the gene activity with disease. However, proteomic technologies allow us to understand proteins and their modifications, elucidating properties of cellular behavior that may not be reflected in analysis of gene expression. The application of these innovative approaches has recently yielded the promising new urinary biomarkers for acute kidney injury and chronic kidney disease, thus providing a better insight in renal pathophysiology and establishing the basis for new therapeutic strategies. Despite significant improvements in therapeutics, the mortality and morbidity associated with acute renal failure (ARF) remain high. The lack of early markers for ARF causes an unacceptable delay in initiating therapy. These biomarker panels will probably be useful for assessing the duration and severity of ARF, and for predicting progression and adverse clinical outcomes. Kidney failure leads to the uremic syndrome characterized by accumulation of uremic toxins, which are normally cleared by the kidneys. Proteomics has gained considerable interest in this field, as a new and promising analytical approach to identify new uremic toxins. The urinary proteome as a tool for biomarker discovery is still in its early phase. A major challenge will be the integration of proteomics with genomics data and their functional interpretation in conjunction with clinical results and epidemiology. PMID- 19814649 TI - Effects of emissions from different type of residential heating upon cyclic guanosine monophosphate (cGMP) in blood platelets of residents. AB - We hypothesized that different types of residential heating would be associated with different levels of indoor carbon monoxide (CO) and further that this might result in a differential in the concentration of cyclic 3':5' guanosine monophosphate (cGMP) in blood platelets in exposed residents. Individuals, who were recruited from homes using different fuel for heating, donated a venous blood sample in the winter and in the summer. In the winter the median blood platelet cGMP value for the group using liquid propane gas (LPG) was 65% higher than for the group using piped natural gas for heating (p <0.001). Also in the group using LPG, the median concentration of cGMP in the winter was 39% higher than the summer median (p < 0.003). The mean indoor concentrations of CO were measured over a period of 1 week during the winter and were <1 ppm. We conclude that observed differences were associated with emissions from different types of heating but that CO exposure alone is too low to explain these. PMID- 19814653 TI - Health-based Provisional Advisory Levels (PALs) for homeland security. AB - The Homeland Security Presidential Directive #8 (HSPD-8) for National Emergency Preparedness was issued to " establish policies to strengthen the preparedness of the United States to prevent and respond to threatened or actual domestic terrorist attacks, major disasters, and other emergencies by requiring a national domestic all- hazards preparedness goal. "In response to HSPD-8 and HSPD-22 (classified) on Domestic Chemical Defense, the US Environmental Protection Agency (US EPA) National Homeland Security Research Center (NHSRC) is developing health based Provisional Advisory Levels (PALs) for priority chemicals (including chemical warfare agents, pesticides, and toxic industrial chemicals) in air and drinking water. PALs are temporary values that will neither be promulgated, nor be formally issued as regulatory guidance. They are intended to be used at the discretion of risk managers in emergency situations. The PAL Program provides advisory exposure levels for chemical agents to assist in emergency planning and response decision-making, and to aid in making informed risk management decisions for evacuation, temporary re-entry into affected areas, and resumed-use of infrastructure, such as water resources. These risk management decisions may be made at the federal, state, and local levels. Three exposure levels (PAL 1, PAL 2, and PAL 3), distinguished by severity of toxic effects, are developed for 24 hour, 30-day, 90-day, and 2-year durations for potential exposure to drinking water and ambient air by the general public. Developed PALs are evaluated both by a US EPA working group, and an external multidisciplinary panel to ensure scientific credibility and wide acceptance. In this Special Issue publication, we present background information on the PAL program, the methodology used in deriving PALs, and the technical support documents for the derivation of PALs for acrylonitrile, hydrogen sulfide, and phosgene. PMID- 19814654 TI - Inhibition of LH-stimulated androgen production in rat immature Leydig cells: Effects on nuclear receptor steroidogenic factor 1 by FGF2. AB - Both fibroblast growth factor 2 (FGF2) and luteinizing hormone (LH) have been reported to regulate androgen production in Leydig cells in progenitor Leydig cells. The objective of the present study is to examine the regulation of androgen production in rat immature Leydig cells (ILCs). ILCs were isolated from 35-day-old rat testes and cultured in DMEM/F12 medium with LH (1 ng/ml) or FGF2 (10 ng/ml). 5alpha-Androstane-3alpha, 17beta-diol (3alpha-DIOL), the primary androgen in ILCs, and testosterone (T) were measured by Radioimmuno assay. The results showed the LH stimulated androgen production in ILCs, and FGF2 did not. However, FGF2 decreased the LH-stimulated androgen production. Real-time PCR and enzyme assay showed that FGF2 decreased levels of several steroidogenic enzymes, inhibited the expressions of steroidogenic acute regulatory (StAR) protein and steroidogenic factor 1 (Nr5a1) in LH-stimulated ILCs. FGF2-mediated inhibition of Nr5a1gene expression may be the mechanism through which FGF2 inhibits LH stimulated androgen production. PMID- 19814655 TI - Long-term and acute effects of gliadin on small intestine of patients on potentially pathogenic networks in celiac disease. AB - Celiac disease (CD) is a complex, immune-mediated intolerance to gliadin that develops in genetically susceptible individuals. Although the main driving force of the disease is an aberrant autoimmune response, several other pathogenic mechanisms, many still unidentified, are also involved. In order to describe at a network level the alterations provoked by a gliadin insult on the intestinal mucosa of patients, we compared the expression profiles of biopsies from 9 active and 9 treated patients (long-term effects of gliadin), and of 10 biopsies from gluten-free diet treated patients that were incubated in vitro with or without gliadin (acute effects) and integrated significantly altered transcripts into potentially pathogenic biological processes. Using information on Kyoto Encyclopedia of Genes and Genomes pathways and Gene Ontology terms represented among the differentially expressed genes, we observed important dysfunction in several complex networks, including those related to cell-cell communication, intracellular signaling, ubiquitin-proteasome system, cell cycle/apoptosis and extracellular matrix. The reconstruction of the role of these biological networks in the development of the intestinal lesion in CD provides a comprehensive picture of key events that contribute to the disease, and could point towards novel functional candidates that might be potential therapeutic targets or responsible for genetic susceptibility. PMID- 19814656 TI - SR 57746A/xaliproden, a non-peptide neurotrophic compound: prospects and constraints for the treatment of nervous system diseases. AB - Neurodegenerative disorders such as Alzheimer's disease or amyotrophic lateral sclerosis as well as peripheral neuropathies are difficult to treat due to a limited range of effective drugs. Neurotrophic growth factors promote neuronal survival and differentiation and could hence be interesting tools to treat these diseases. Their therapeutic use is limited due their short half-life, their inability to cross the BBB and potential side effects including tumor promotion. SR 57746A is a non-peptide, orally active compound that exhibits neuroprotective effects in various model systems in vitro and in vivo. SR 57746A shows--amongst other activities--agonistic activity on 5-HT(1A) receptors. Several clinical trials examined SR 57746A in patients with Alzheimer's disease, amyotrophic lateral sclerosis or chemotherapy-induced peripheral sensory neuropathy. This article reviews the preclinical and clinical data on SR 57746A and points out potential future applications of this compound. However, due to disapointing results in phase III trials, Sanofi-Aventis recently decided to discontinue the development of this drug. PMID- 19814657 TI - UGT3A: novel UDP-glycosyltransferases of the UGT superfamily. AB - Mammalian UDP-glycosyltransferases (UGTs) are divided into four families: UGT1, UGT2, UGT3, and UGT8. UGT3 is the last of the gene families to be identified, and until relatively recently, little was known about the function of these enzymes. In this article, we present new analyses of the UGT3 family genes, including the structure of the UGT3A locus, interspecies sequence conservation, single nucleotide polymorphisms, and splice variants. We also review recently published work that has revealed that one member of this family, UGT3A1, has a unique enzymatic function: N-acetylglucosaminidation. Finally, we discuss the possible biological significance of the UGT3A enzymes. PMID- 19814658 TI - Synthetic vehicles for DNA vaccination. AB - DNA vaccination is an attractive immunization method able to induce robust cellular immune responses in pre-clinical models. However, clinical DNA vaccination trials performed thus far have resulted in marginal responses. Consequently, strategies are currently under development to improve the efficacy of DNA vaccines. A promising strategy is the use of synthetic particle formulations as carrier systems for DNA vaccines. This review discusses commonly used synthetic carriers for DNA vaccination and provides an overview of in vivo studies that use this strategy. Future recommendations on particle characteristics, target cell types and evaluation models are suggested for the potential improvement of current and novel particle delivery systems. Finally, hurdles which need to be tackled for clinical evaluation of these systems are discussed. PMID- 19814659 TI - Stroke prognosis in diabetes mellitus: new insights but questions remain. AB - Diabetes is not only an independent risk factor for lacunar infarction, especially in patients with multiple lacunar infarcts, but the presence of diabetes is also associated with worse functional recovery in these patients. PMID- 19814660 TI - Cellular stroke therapy: from cell replacement to trophic support. AB - Although initially considered as primarily a cell-replacement strategy, researchers today attribute a combination of both cellular repair and trophic support from transplanted cells as mediators of recovery. PMID- 19814661 TI - PGI(2) receptor agonist demonstrates efficacy in pulmonary arterial hypertension. AB - The first orally active nonprostanoid PGI(2) receptor agonist has shown highly positive results in a Phase IIa study for the treatment of pulmonary arterial hypertension. PMID- 19814662 TI - Cangrelor: a review on its mechanism of action and clinical development. AB - In patients with acute coronary syndromes and undergoing percutaneous coronary intervention, numerous large-scale clinical trials have shown that adjunctive treatment with the P2Y(12) receptor antagonist clopidogrel in addition to aspirin reduces ischemic events. These studies underscore the importance of blockade of the P2Y(12) signaling pathway in these settings. However, recent findings have shown that clopidogrel therapy may have some shortcomings. These include its broad range of interindividual-response profiles, where patients with low P2Y(12) inhibitory effects have an increased risk of recurrent ischemic events, including stent thrombosis, and its irreversible mechanism of action. These observations underscore the need for novel antiplatelet agents overcoming these limitations. Cangrelor (AR-C69931MX) is an intravenous, direct-acting and reversible P2Y(12) receptor antagonist. Cangrelor has a rapid onset and offset of action and achieves significantly greater degrees of platelet inhibition compared with clopidogrel. This article provides an overview of the current status of knowledge on cangrelor, focusing on its pharmacologic properties, clinical development and potential future applications. PMID- 19814663 TI - Efficacy of aspirin for secondary prevention in patients with peripheral artery disease. AB - Evaluation of: Berger JS, Krantz MJ, Kittelson JM, Hiatt WR. Aspirin for the prevention of cardiovascular events in patients with peripheral artery disease: a meta-analysis of randomized trials. JAMA 301, 1909-1919 (2009). Aspirin decreases the risk of cardiovascular events in patients with prior coronary heart or cerebrovascular disease. The American College of Cardiology/American Heart Association guidelines recommend a low-dose aspirin regimen (75-325 mg/day) to reduce the risk of cardiovascular events in patients with peripheral artery disease (PAD). However, the effect of aspirin for secondary prevention in patients with PAD has not been well established. The paper under evaluation performed a meta-analysis of 18 trials to investigate the effect of aspirin on cardiovascular events (nonfatal myocardial infarction, nonfatal stroke and cardiovascular death) in patients with PAD. The results of this meta-analysis in a PAD cohort revealed that treatment with aspirin did not significantly reduce the combined end point of cardiovascular events; however, aspirin resulted in a significant reduction in the incidence of nonfatal stroke. This analysis raises a number of questions regarding the overall efficacy of aspirin in PAD and what should be the optimal antiplatelet therapy in patients with PAD: aspirin, clopidogrel or perhaps a combination of aspirin and clopidogrel. PMID- 19814664 TI - Anticoagulation for heart failure: selecting the best therapy. AB - Heart failure (HF) incidence approaches 10 per 1000 population after the age of 65 years, and is the most common Medicare diagnosis. The management of HF patients has improved over the last decade, but the risk of thromboembolic complications is frequently underestimated and currently, no treatment options have been formally implemented to decrease the incidence of events. In particular, stroke is a major complication of HF. HF ranks second after atrial fibrillation in the number of cardiogenic strokes caused. The actual rate of stroke in recent HF trials ranges between 1.5 and 2.4% per year. Warfarin is of proven efficacy in patients with ischemic heart disease and atrial fibrillation, but there has been little research testing antithrombotic agents in patients with HF. Aspirin may be another promising drug, but trials have shown a higher rate of HF admissions with aspirin, suggesting that aspirin may attenuate the angiotensin converting enzyme inhibitor effect on cardiac function. We eagerly await the completion of large, randomized trials to help guide us in these important issues affecting a large number of patients. PMID- 19814665 TI - Statin use before and after acute ischemic stroke onset improves neurological outcome. AB - The devastating consequences of stroke, both in terms of morbidity and mortality, and the economic implications it has worldwide, coupled with a growing knowledge regarding its pathophysiology, has led to the development of novel therapeutic strategies in recent years that have had an enormous impact on clinical practice and the outcome of stroke patients. While thrombolysis is regarded as the most important of these developments, its relatively narrow therapeutic window and complexity of administration imply that only a minority of stroke patients can benefit from it. Among other strategies, in an attempt to overcome this limitation, research has focused on the development of neuroprotection aimed to salvage ischemic brain tissue by means complementary to reperfusion. Among these approaches, the use of statins stands out, which, by inhibiting the HMG CoA reductase, interfere with the formation of isoprenoid intermediates in the biosynthesis of cholesterol, and have been shown to have a positive impact in both the incidence and outcome of acute stroke. The main results of animal models, observational studies, clinical trials and opinion articles available in the medical literature covering the effect of statins on acute brain ischemia will be reviewed in order to provide in-depth information regarding their mode of action and current evidence regarding their potential benefits for ischemic stroke patients. PMID- 19814666 TI - Statins and stroke prevention. AB - Over the past decade, statins have been proved to significantly decrease coronary events in the primary and secondary prevention of coronary artery disease. Recent clinical trials have indicated that statins significantly reduce stroke risk in patients with vascular disease. A meta-analysis of randomized trials of statins in combination with other preventive strategies, involving 165,792 individuals, showed that each 1-mmol/l (39 mg/dl) decrease in LDL-cholesterol equates to a reduction in relative risk for stroke of 21.1 (95% CI: 6.3-33.5; p = 0.009). It is not known whether these findings might be due to the cholesterol-reduction effect of statins or to the pleiotropic effects of statins, such as improved endothelial function, decreased platelet aggregability and reduced vascular inflammation. In the secondary prevention of stroke, The Stroke Prevention by Aggressive Reduction of Cholesterol Levels study found that treatment with atorvastatin reduced the risk of recurrent cerebrovascular events in patients with recent stroke or transient ischemic attack but no history of heart disease. PMID- 19814667 TI - Antithrombotic medication for stroke prevention. AB - This article provides the reader with an overview and an update on antithrombotic stroke-prevention strategies. Relevant journals were hand-searched by the authors to compile a broad, but not comprehensive, summary of innovative and clinically relevant studies. These findings were then summarized to provide an expert opinion on selected fields of medical stroke prevention. We conclude that aspirin, clopidogrel or dipyridamol-aspirin are the cornerstone therapies in patients with noncardioembolic stroke. More potent antiplatelet drugs or the combination of aspirin and clopidogrel prevent more ischemic events but also lead to more bleeding complications. For secondary stroke prevention in patients with atrial fibrillation, oral anticoagulation is more effective than aspirin or the combination of aspirin and clopidogrel. PMID- 19814668 TI - Ischemic tolerance in stroke treatment. AB - Although outcome after stroke treatment has significantly improved over the last 30 years, there has been no revolutionary breakthrough. Among different combined approaches, systemic thrombolysis in combination with neuroprotection became a favorite research target. Recent studies suggest that transient ischemic attacks may represent a clinical model of such ischemic tolerance; thus, a new focus on this research has emerged. In this review, we show the parallels between ischemia and neuroprotection and discuss the potential therapeutic options that may be opened by this new molecular knowledge. PMID- 19814669 TI - Stroke in HIV infection and AIDS. AB - HIV/AIDS appears to increase the risk of both ischemic and hemorrhagic stroke. This increased risk is most apparent in the young HIV-infected population in which other risk factors for stroke are seldom evident. Mechanisms underlying the increased risk include opportunistic infectious meningitides and vasculitides, primary HIV vasculopathy, altered coagulation and cardioembolic events, although the cause may be multifactorial or remain cryptic. With better control of HIV via effective, highly active antiretroviral therapy, the role of many of these risks has been mitigated, only to be supplanted by an aging population with more conventional atherosclerotic risk factors magnified by the hyperlipidemia attending the use of protease inhibitors. Selecting the appropriate therapy for treating stroke in the HIV-infected patient is dependent on diagnostic rigor in identifying its underlying etiology. PMID- 19814670 TI - Predicting recurrent stroke after minor stroke and transient ischemic attack. AB - The risk of a subsequent stroke following an acute transient ischemic attack or minor stroke is high, with 90-day risk at approximately 10%. Identification of those patients at the highest risk for recurrent stroke following a transient ischemic attack or minor stroke may allow risk-specific management strategies to be implemented, such as hospital admission with expedited work-up for those at high risk and emergency room discharge for those at low risk. Predictors of recurrent stroke, including the ABCD2 score, brain imaging and the stroke mechanism, are reviewed in this article, with a focus on recent literature. An emphasis is placed on the importance of early imaging of the brain parenchyma (diffusion-weighted imaging) and vascular imaging to identify patients at high risk for recurrence. The need for identification of the cause of the initial event, allowing therapies to be tailored to the individual patient, is discussed. PMID- 19814671 TI - Chronic total occlusions in peripheral vasculature: techniques and devices. AB - Peripheral artery disease continues to increase in an ever-aging society and consumes a significant part of health resources. Chronic total occlusions account for a significant portion of the lesions encountered by endovascular interventionalists. Unlike the coronary circulation, these occlusions are often long and associated with other features of complexity. The two primary issues concerning these lesions are the ability to safely achieve intimal angiographic success and the long-term durability of therapy. Traditionally, the use of percutaneous techniques was limited to certain anatomic subsets, such as stenosis or total occlusions. However, recent advances in wire technology, re-entry catheters, alternative crossing tools and atherectomy devices have increased the interventional armamentarium for this challenging disease subset. This article will focus on some of the techniques in crossing chronic total occlusions lesions, as well as the available devices to improve long-term patency. PMID- 19814673 TI - The proceedings of the 17th International Conference on Chelation: application of effective chelation therapies in iron loading and non iron loading conditions, and the gap in the prevention and treatment policies on thalassemia between developed and developing countries. AB - Substantial progress in the use of chelating drugs for the treatment of iron overload and of non iron loading conditions has been presented during the 17th International Conference on Chelation (ICOC) held in November 2007 at Shenzhen, China. Major challenges lie ahead for the prevention and treatment of thalassemia in China, India, Thailand, Indonesia and many other developing countries where millions of heterozygote thalassemia carriers live and thousands of homozygote thalassemia patients are born annually. The progressive improvement of the economic climate in developing countries will increase the demand and resources for more prenatal and antenatal diagnoses, transfusions and chelation therapy in forthcoming years. Despite the major advances in diagnosis and treatment in developed countries, the vast majority of thalassemia patients in developing countries die untreated because they cannot afford the cost of transfusions and chelation therapy. New approaches and infrastructures and more efforts are needed to overcome the difficulties of supplying new techniques and treatments to patients in developing countries. International and local organizations need to be persuaded to act collectively and effectively to improve chelation and related treatments for thalassemia and other conditions, especially at this time that universally effective and inexpensive chelation therapies can be applied. PMID- 19814674 TI - Advances in the prevention and treatment are changing thalassemia from a fatal to a chronic disease. experience from a Cyprus model and its use as a paradigm for future applications. AB - Thalassemia is endemic in Cyprus with a frequency of 1 in 6 persons being a heterozygote and about 1 in 1,000 a homozygous thalassemia major patient. Cyprus has been a pioneer nation in reducing and almost eliminating the number of births of thalassemia major patients by introducing prenatal and antenatal diagnosis. The risks associated with bone marrow transplantation (BMT) make transfusion and chelation therapy the major form of treatment for the vast majority of thalassemia patients. Improved transfusion techniques, diagnostic methods, iron chelation and supportive therapy have increased the quality of life and survival of patients, some of whom are exceeding 50 years of age. The introduction of effective chelation therapy protocols using primarily deferiprone (L1) in combination with deferoxamine (DFO) resulted in the reduction of iron overload induced cardiac failures, which is the main cause of death in thalassemia major. Despite their chronic condition and tedious clinical management many patients are successful professionals, married and have children. The advancement in treatment is transforming thalassemia from a fatal to a chronic condition and some families are opting for giving birth to a thalassemic child rather than abortion. PMID- 19814675 TI - Thalassemia incidence and treatment in China with special reference to Shenzhen City and Guangdong province. AB - According to the data from different screening studies, thalassemia is the most commonly seen hereditary hemolytic disease in China. The reported prevalence of thalassemia carriers varies but it is most prevalent in Southern China. In the past, the outcome of patients with thalassemia major has been very poor due to unfavorable economic background. With economic improvement in the past 10 years, increasing number of patients can get regular transfusion and chelation, and more patients can be treated by hemopoietic stem cell transplantation. A better prevention network has been built up over the years, but there are still babies being born with severe forms of thalassemia every year. A more comprehensive preventive program and public education are vital. PMID- 19814676 TI - Update on thalassemia treatment in Taiwan, including bone marrow transplantation, chelation therapy, and cardiomyopathy treatment effects. AB - Over the past few decades, Taiwan has seen striking improvements in the life expectancy of its 400 registered beta-thalassemia major (beta-TM) patients due mainly to adequate transfusion regimens and effective iron chelation therapy. Since 1995, Taiwanese citizens have enjoyed universal health care through National Health Insurance (NIH), receiving comprehensive treatment at minimal cost. In 1984, a national program for thalassemia prevention, control, and hematopoietic stem cell transplantation (HSCT) was initiated. Recent data show 1- and 2-year event-free survival rates of 85 and 78%, respectively. Chelation agents like deferoxamine (DFO), deferiprone (L1) and deferasirox (DFRA) are available in Taiwan, and therapy is tailored to individuals based on drug availability and tissue distribution of iron load. Intensive chelation regimens combining L1 and DFO are recommended in patients with cardiac complications, while DFRA has been found to be effective in reducing serum ferritin, with acceptable side effects. Here, we report advances in thalassemia treatment in Taiwan and suggest treatment guidelines. PMID- 19814677 TI - Uses and limitations of serum ferritin, magnetic resonance imaging T2 and T2* in the diagnosis of iron overload and in the ferrikinetics of normalization of the iron stores in thalassemia using the International Committee on Chelation deferiprone/deferoxamine combination protocol. AB - Excess cardiac iron deposition leads to congestive cardiac failure and accounts for more than 70% of deaths in thalassemia major patients. In three separate studies involving 145 thalassemia patients, serum ferritin and magnetic resonance imaging (MRI) relaxation times T2 and T2* have been compared for assessing iron load levels during chelation treatment. In two studies, variable levels of cardiac iron load have been detected by T2 and T2* in patients treated with deferoxamine (DFO), which, however, were unrelated to serum ferritin. In most cases, similar range levels from normal to severe cardiac iron load could be identified by both the T2 and T2* methods. However, in a few cases there were substantial differences in the levels detected between the two methods. In the third study, the ferrikinetics of the normalization of the iron stores during the International Committee on Chelation (ICOC) deferiprone (L1)/DFO combination protocol was followed up using T2 and T2* and serum ferritin. Iron deposits were found not to be proportionally distributed between the liver and the heart or uniformly distributed within each organ. Iron mobilization in each patient varied and iron deposits in each organ were cleared at different rates. Despite some limitations, the application of the MRI relaxation times T2 and T2* offers the best diagnostic methods for iron overload estimations in most organs and especially the heart. These MRI methods and serum ferritin could also be used for the ferrikinetics of iron mobilization and removal during chelation therapy and the normalization of the iron stores during the ICOC L1/DFO combination protocol. There is a need to standardize the two MRI relaxation times T2 and T2* methods and identify the factors causing the differences between them. PMID- 19814679 TI - A new era in iron chelation therapy: the design of optimal, individually adjusted iron chelation therapies for the complete removal of iron overload in thalassemia and other chronically transfused patients. AB - A new era in iron chelation therapy began with the successful removal of excess iron load and the maintenance of normal iron stores in thalassemia patients using the International Committee on Chelation (ICOC) protocols. This achievement was based on two phases, firstly the introduction of deferiprone (L1) (80-100 mg/kg/day) and deferoxamine (DFO) (40-60 mg/kg at least 3 days per week) combination therapy, which appears to progressively remove all excess storage iron and thereafter by the introduction of L1 monotherapy that can maintain physiological range levels of serum ferritin, cardiac and liver magnetic resonance imaging (MRI) T2*. This new development is likely to change current practices and set a new gold standard in the treatment of transfusional iron loaded patients leading to an increased survival and the change of thalassemia from a fatal to a chronic disease. A major aspect of the improved therapies is the ability of L1 to mobilize and remove excess cardiac iron and reduce congestive cardiac failure, which is the main cause of death in thalassemia patients. Further, new developments include the use of alternating sequential chelation therapies and selected dose protocols with L1, DFO and deferasirox (DFRA) for overcoming toxicity and efficacy complications observed in some patients treated with monotherapies or combination therapies. The selection and adjustment of dose protocols is crucial for providing optimum chelation therapy for each individual patient. PMID- 19814678 TI - A practical chelation protocol based on stratification of thalassemic patients by serum ferritin and magnetic resonance imaging cardiac T2*. AB - Our previous study showed that combined therapy with deferiprone (L1) and deferoxamine (DFO) was safe and efficacious in reducing iron overload in poorly chelated thalassemia major patients for the short-term but the magnetic resonance imaging (MRI) T2* evaluation was not available at that time. Since October 2006, we applied a standardized chelation protocol by stratifying transfusion-dependent thalassemic patients into three groups, namely well-chelated group (A), poorly chelated group without (B) or with (C) risk of cardiac complications, based on their serum ferritin (SF) levels and magnetic resonance imaging (MRI) cardiac T2* measurements. The patients in each group were given options of chelation regimens to improve their iron overload status. Chelation regimens included continuation or intensification of DFO alone (Regimen Ic or Ii, respectively), L1 alone (Regimen II), and combined therapy with L1 and DFO (Regimen III). Group A patients continued with Regimen Ic. Group B patients could opt for either Regimen Ii or II/III. Group C patients could opt for either Regimen Ii or III. Serum ferritin levels and MRI cardiac and liver T2* measurements were evaluated after 1 year of treatment. Fifty-seven patients (27 males, 30 females; age range 5-34 years, median: 25 years) were categorized into Group A (n = 3), B (n = 20) and C (n = 34). All Group A patients continued with DFO treatment. In Group B, seven were on Regimen Ii, five on Regimen II and five on Regimen III. In Group C, five were on Regimen Ii, two on Regimen II and 26 on Regimen III. Significant improvement was noted only for Group C patients using Regimen III (combined therapy) in SF levels, cardiac T2* and liver T2* measurements. PMID- 19814680 TI - Iron chelation therapy for patients with myelodysplastic syndrome. AB - Chronic blood transfusions are necessary for patients with hereditary anemia such as thalassemia, and for patients with myelodysplastic syndrome (MDS) who become anemic and transfusion-dependent. A common consequence of chronic transfusion is iron accumulation that can lead to organ damage. While there is general agreement regarding the value of iron chelation therapy to reduce the detrimental effects of iron overload in thalassemia major, the same is not true for MDS. The malignant nature of the disease and the relatively high cost of iron chelation therapy make cost-effectiveness an issue of great concern. Furthermore, the positive assessment of a drug's cost-effectiveness in one country does not necessarily justify its use in another country. More prospective studies are needed to identify the best iron chelator for patients with MDS as well as to identify those patients who will benefit most from iron chelation therapy. PMID- 19814681 TI - Treatment with deferiprone for iron overload alleviates bone marrow failure in a Fanconi anemia patient. AB - Fanconi anemia (FA) is a rare inherited disorder characterized by congenital abnormalities, progressive bone marrow failure and cancer susceptibility. There are no reports in the literature about a specific therapy effective in treating the progressive bone marrow failure of FA except for hematopoietic stem cell transplantation (HSCT). A FA patient started to receive deferiprone (L1) therapy due to iron overload. We report here that the white blood cell counts, hemoglobin (Hb) levels and platelet counts were significantly higher during the L1-treated period than when without L1 therapy. Therefore, L1 therapy may be worth considering for FA patients who cannot undergo HSCT. PMID- 19814682 TI - Effects of chelators (deferoxamine, deferiprone and deferasirox) on the growth of Klebsiella pneumoniae and Aeromonas hydrophila isolated from transfusion dependent thalassemia patients. AB - Infections are among the leading causes of death for thalassemia major patients. The known predisposing factors of infection include prior splenectomy, iron overload and use of iron chelator such as deferoxamine (DFO). While encapsulated organisms frequently found in splenectomized patients were readily controlled by prophylactic vaccination and vigilant antibiotic treatment, ferrophilic organisms such as Yersinia and Klebsiella remain common pathogens in thalassemic patients. Yersinia infections are more prevalent in temperate regions and Klebsiella infections are commonly found in tropical and subtropical areas. While the use of DFO further aggravates the risk of Yersinia infection, oral chelators such as deferiprone (L1) do not enhance the growth of Yersinia in vitro or in vivo. We found that the growth of Klebsiella was marginally enhanced by DFO in vitro when compared to Yersinia. Such an unfavorable effect was not found in either L1 or deferasirox (DFRA) in vitro. The growth of Aeromonas was not affected by the presence of all three forms of chelators. Therefore, we suggest that factors other than DFO may account for the increased prevalence of Klebsiella and Aeromonas infection in Asian thalassemic patients. PMID- 19814683 TI - Long-term effects of pamidronate in thalassemic patients with severe bone mineral density deficits. AB - Osteoporosis is a common complication in thalassemia major (TM). Our previous study demonstrated severe bone mineral density (BMD) deficits at spine and hip in 62 and 35% of TM patients. This study assessed the effects of different treatments (calcium, vitamin D and bisphosphonate) on patients' BMD, which was measured at baseline and after 3-year treatments by dual energy X-ray absorptiometry (DEXA). Twenty-one untreated patients, 11 patients on calcium/vitamin D and seven patients on additional pamidronate, were recruited. They were comparable for gender (p = 0.630) and serum ferritin levels (p = 0.412). The median BMD Z-scores at lumbar spine and left hip improved only in patients with standard plus pamidronate treatments (baseline: -3.01 and -3.05, end-of-study: -2.12 and -2.09; p = 0.018 and 0.028, respectively). In contrast, BMD Z-scores at hip worsened in untreated patients (p = 0.034). In conclusion, long-term improvement in BMD in TM patients was observed with bisphosphonate but not calcium and vitamin D treatment. PMID- 19814684 TI - Risk/benefit assessment, advantages over other drugs and targeting methods in the use of deferiprone as a pharmaceutical antioxidant in iron loading and non iron loading conditions. AB - Tissue damage caused by oxidative stress is a common characteristic of many conditions involving different major organs such as the brain, heart, liver and kidneys. The treatment of such conditions using classical antioxidants is not in most cases sufficient or effective because it lacks specificity and has a low therapeutic index. Increased evidence from in vitro, in vivo and clinical studies suggest that deferiprone (L1) can be used as a potent pharmaceutical antioxidant by mobilizing labile iron and copper and/or inhibiting their catalytic activity in the formation of free radicals and oxidative stress in tissue damage. The high therapeutic index, tissue penetration, rapid iron binding and clearance of the iron complex, and the low toxicity of L1, support its application as an antioxidant pharmaceutical for adjuvant, alternative or main therapy, especially in conditions where other treatments have failed. Substantial clinical improvement and reversal in most cases of the tissue damage has been observed in cardiomyopathy in thalassemia, diabetic nephropathy and glomerulonephritis in kidney disease, Friedreich's Ataxia and Fanconi Anemia patients. In contrast to L1, both deferoxamine (DFO) and deferasirox (DFRA) have major disadvantages in their use in non iron loading conditions due to toxicity implications. Further studies in the above and other conditions and optimization of the L1 therapy in each individual will increase the prospects of the application and role of L1 as a universal antioxidant pharmaceutical. PMID- 19814685 TI - Enzyme levels and G-463A polymorphism of myeloperoxidase in chronic lymphocytic leukemia and multiple myeloma. AB - The aim of this study was to investigate how myeloperoxidase (MPO) G-463A gene polymorphism and enzyme levels varied among patients with chronic lymphocytic leukemia (CLL) and multiple myeloma (MM) and to find the relationship between the MPO gene, enzyme levels, and clinical parameters. We studied the sera from 40 healthy volunteers, patients with CLL (n = 34) and MM (n = 28). In subjects with homozygote GG genotype, MPO levels were higher in the patients with both CLL and MM than in the control group. This difference was statistically significant in patients with CLL. In conclusion, homozygote GG genotype is found to be associated with an increasing amount of serum MPO. In accordance with the results of the study, we assess that the increase in the MPO enzyme level in the patient groups with CLL and MM generated bactericidal effects as well as the increased formation of ROP, thus setting off a pro-cell death pathway and playing a role on the pathogenesis of lymphoproliferative malignancies through this mechanism. PMID- 19814686 TI - Decreased major histocompatibility complex class II expression in diffuse large B cell lymphoma does not correlate with CpG methylation of class II transactivator promoters III and IV. PMID- 19814687 TI - Cyclophosphamide, doxorubicin, vincristine, methotrexate, bleomicin and prednisone plus rituximab in untreated young patients with low-risk (age-adjusted international prognostic index 0-1) diffuse large B-cell lymphoma. AB - Young patients (aged 18-60 years) with good-prognosis diffuse large B-cell lymphoma (DLBCL) [0 and 1 risk factor according to age-adjusted international prognostic index (aa-IPI)] are distinguished from patients with poor-prognosis. Six cycles of cyclophosphamide, doxorubicin, vincristine, prednisone (CHOP) in combination with rituximab achieved best results in young patients with low-risk DLBCL. We retrospectively analyzed the data of 82 patients (18-60 years) with untreated aa-IPI 0-1 DLBCL, from six Italian institutions, who underwent, between January 2002 and January 2007, rituximab-cyclophosphamide, doxorubicin, vincristine, methotrexate, bleomicin and prednisone (R-MACOP-B) therapy followed, in patients with bulky presentation, by 30-36 Gy involved-field radiation therapy (34 patients). An overall response rate was noted in 77 out of 82 (94%) patients (75 patients had a complete response (91%), 2 patients had a partial response). With a median follow-up of 46 months, 7-year progression-free and overall survival were estimated to be 91% and 94%, respectively. R-MACOP-B regimen followed by involved-field radiation on bulky presentation is safe and very effective in the treatment of young patients with low-risk DLBCL. PMID- 19814688 TI - Loss of Irf8 does not co-operate with overexpression of BCL-2 in the induction of leukemias in vivo. PMID- 19814689 TI - Dedication to the clinical investigators for their work with hairy cell leukemia. PMID- 19814690 TI - Historical aspects and milestones in the development of effective treatment for hairy cell leukemia. AB - Hairy cell leukemia (HCL) is a rare B-cell lymphoproliferative disorder, but is gratifying to treat for both physicians and patients. During the 50 years since its initial description as a clinical entity, hematologists have been fascinated by the bizarre appearance of the malignant cell with its hair-like projections. Therapeutic strategies have evolved from splenectomy to interfere to the purine analogues which have become the treatments of-choice and are very effective. Immunoconjugate Therapy with BL22 (anti-CD22 [corrected] antibody linked to truncated to Pseudomonas exotoxin) represents the newest milestone in the development of effective treatment for hairy cell leukemia. PMID- 19814691 TI - The biology of hairy cell leukemia. AB - As in all malignancies, the biology of HCL reflects both the behaviour of the malignant cells (hairy cells) themselves and their two-way interaction with the microenvironment. However, the tissue interactions of HCs are particularly striking and involve extensive remodelling of bone marrow, spleen and liver, with relative sparing of lymph nodes. The mechanistic basis of this remodelling is now largely understood and is described herein. Regarding HCs themselves, they are late clonal B cells which have often undergone heavy-chain-isotype class switching and whose VH genes are usually mutated. HCs are highly activated cells in which a number of signalling pathways are constitutively active. This activation determines many of the specific features of HCs, but its cause remains unknown. PMID- 19814692 TI - Cladribine in the treatment of hairy cell leukemia: initial and subsequent results. AB - Hairy cell leukemia (HCL) is a chronic B-cell lymphoproliferative disorder associated with pancytopenia, splenomegaly, and recurrent infections. Although interferon alpha and pentostatin were initially found to be effective in this disease, cladribine has emerged as the preferred initial therapy. Cladribine given as a single continuous intravenous seven-day infusion is the dosing schedule with the most durable complete remissions and evaluated in the greatest number of patients with HCL. Patients who relapse after purine analogue therapy, whether it be cladribine or pentostatin, can be successfully retreated with cladribine. Patients with HCL may develop complications of recurrent infection and second malignancies. Although both complications are postulated to be related to therapy, they may also be due to the duration and burden of the disease. Minimal residual disease detected on bone marrow biopsy is thought to predict for future relapse. We will review the initial and subsequent results of cladribine in the management of HCL. PMID- 19814693 TI - Combination therapies to improve the long-term outcome in hairy cell leukemia. AB - Since its first description in 1958, hairy cell leukemia (HCL) had a long period of different unsatisfactory treatments till the early 1980s, when interferon-alfa and subsequently purine analogs (PAs) pentostatin or cladribine, or pentostatin were described to determine durable responses. However, second lines of treatments are very often needed in the life span of patients with HCL and a small minority of patients are refractory and with poor prognosis. These disadvantages have led to test the efficacy of additional compounds, mainly monoclonal antibodies, combined with PAs. The objective of this presentation is to review the current knowledge of combination treatments in HCL. Combination treatments have been used or proposed for different purposes in HCL: to reduce acute toxicity, to improve complete remission (CR) rate, to overcome resistance/refractoriness, to prolong disease-free survival, or to prolong overall survival. The type of patients to be selected for the combinations and the perspectives of combinations of PAs with monoclonal antibodies will also be discussed. PMID- 19814694 TI - Long-term follow-up studies in hairy cell leukemia. AB - Enormous progress has been made in the management of patients with hairy cell leukemia (HCL) over the past 50 years since this disease was initially described in 1958. The introduction of the two commonly used purine nucleoside analogs (pentostatin and cladribine, respectively) has independently changed the natural history of this rare malignancy. Both agents are equivalent in terms of response and long-term results. Advances in therapy are being further pursued with inclusion of monoclonal antibodies (e.g. rituximab) and other immunotherapeutic approaches. Patients with this disease now can live a near normal life expectancy, but the disease has not yet been cured. Clinical trials must continue to address the remaining unanswered questions. PMID- 19814695 TI - Importance of minimal residual disease in hairy cell leukemia: monoclonal antibodies as a therapeutic strategy. AB - With the use of nucleoside analogs as frontline therapy, the prognosis of hairy cell leukemia (HCL) has improved dramatically. Unfortunately, disease recurrence remains problematic. Eradication of minimal residual disease (MRD) persisting after therapy may further improve outcome. The evolution of available techniques used to assess MRD, and the potential incorporation of novel agents such as monoclonal antibodies (MoAbs) into the treatment armamentarium for HCL mandate that MRD analyses be performed concurrently with routine assessments of disease status. Herein, the available data regarding the prevalence and clinical relevance of MRD after therapy for HCL is reviewed. PMID- 19814696 TI - Approach to the patient after relapse of hairy cell leukemia. AB - The current hairy cell leukemia (HCL) treatment is excellent, but evidence of cure with purine analogs cladribine and pentostatin, is lacking. Significant long term immune suppression, particularly to CD4+ T-cells, and declining complete remission rates with each course, make the identification of new therapies an important goal. The anti-CD20 monoclonal antibody (Mab) rituximab displays significant activity, and, while causing prolonged normal B-cell depletion, spares T-cells. Recombinant immunotoxins, containing an Fv fragment of a Mab fused to truncated Pseudomonas exotoxin, have shown efficacy in HCL resistant to both purine analogs and rituximab. LMB-2 targets CD25 and can induce remission providing the HCL cells are CD25+. All HCL cells display CD22. Recombinant anti CD22 immunotoxin BL22, targeting CD22, has shown significant efficacy in phase I and II testing, and avoids prolonged suppression of both normal B- and T-cells. An improved high-affinity version of BL22, termed HA22, is currently undergoing phase I testing. PMID- 19814697 TI - Treatment outcomes for percutaneous nephrolithotomy in renal allografts. AB - PURPOSE: We reviewed our experience with percutaneous nephrolithotomy (PCNL) in patients with renal allografts over 19 years. MATERIALS AND METHODS: We cross referenced all renal allograft recipients at our institution with all PCNLs performed. We identified 16 renal allograft recipients who underwent a total of 21 PCNL procedures between 1986 and 2005. These charts were abstracted and the outcomes reviewed. RESULTS: The mean follow-up was 7.2 years. No loss of renal function occurred in the perioperative period. Five patients lost graft function during the follow-up period; however, none of these patients' loss of function was related to the PCNL. No patient had sepsis. Thirteen of the 16 patients were rendered stone free during long-term follow-up. CONCLUSIONS: PCNL is a safe and effective method of treating nephrolithiasis in a transplanted kidney. PMID- 19814698 TI - Clinical experience with ultrasound-based real-time tracking lithotripsy in the single renal stone treatment. AB - PURPOSE: An ultrasound-based real-time tracking system has been shown to improve the efficacy of lithotripsy in animal studies. This system has been implemented into an electromagnetic shockwave lithotripter (LM-9200 ELMA), and the treatment outcomes were prospectively evaluated. MATERIALS AND METHODS: A total of 1332 patients with one renal stone were treated with LM-9200 ELMA lithotripter between December 2004 and November 2007. Fluoroscopy and ultrasound were alternately used for calculus localization and focusing first. After the calculus was focused and displayed clearly by ultrasound, the ultrasound-based real-time tracking system was applied for calculus monitoring and shockwave trigger control. All patients were followed for up to 3 months. Data regarding stone location, stone size, shockwave number, treatment time, stone free rate, and possible complications were obtained. RESULTS: Of the 865 males and 467 females treated (mean stone size = 12.3 mm), the stone-free rate in 3 months was 80.0%. For all patients, neither anesthetics nor analgesics were necessary. The incidence of blood pressure and microscopic hematuria grading increment at 2 weeks after treatment was 6.8% and 13.8%, respectively. There were no subcapsular hematoma or prolonged hematuria observed. Stone size was an important factor predicting stone-free rate (9.1 +/- 2.1 mm in the successful group vs. 22.0 +/- 3.1 mm in the failed group, p < 0.001). The calculated efficiency quotient of this machine was 0.67. CONCLUSION: An electromagnetic shockwave lithotripter using ultrasound-based real-time tracking system showed efficient crushing of renal calculi in vivo. Our experience has shown that patients treated by this lithotripter are safe with slight microscopic hematuria and having 80% chance of stone clearance. PMID- 19814699 TI - The effects of tolterodine extended release and alfuzosin for the treatment of double-j stent-related symptoms. AB - AIM: To evaluate the effects of tolterodine extended release (ER) and alfuzosin for the treatment of Double-J stent-related lower urinary tract symptoms. MATERIALS AND METHODS: Fifty-two patients (33 men and 19 women; mean age 52.0 years) who underwent insertion of a Double-J stent after urological surgery were prospectively randomized into three groups. Group 1 included 20 patients who received 10 mg of alfuzosin, once daily for 6 weeks; group 2 included 20 patients who received 4 mg of tolterodine ER, once daily for 6 weeks; group 3 included 12 patients who received a placebo for the same protocol. All patients completed a validated Ureteral Stent Symptom Questionnaire at 6 weeks after the stent placement. RESULTS: The mean urinary symptom index was 22.1 in group 1, 22.1 in group 2, and 28.1 in the placebo group (p = 0.032). The mean pain scores were 8.2, 11.7, and 16.2, respectively (p = 0.020). There were no significant differences in urinary symptoms and pain between the alfuzosin and tolterodine ER groups. In addition, there was no significant difference in the general health, work performance, and sexual performance scores among the groups. CONCLUSIONS: Tolterodine ER and alfuzosin improve stent-related urinary symptoms and body pain. PMID- 19814700 TI - Retroperitoneal laparoendoscopic single-site surgery in urology: initial experience. AB - PURPOSE: To present our initial experience with single-port laparoscopic urologic surgery via the retroperitoneal approach using the Alexis wound retractor with flexible laparoscopic instrumentation. PATIENTS AND METHODS: Since December 2008, 14 patients have undergone single-port laparoscopic surgery for various urological diseases, including renal cyst decortication in 5 patients, adrenalectomy in 2 patients, nephroureterectomy in 2 patients, nephrectomy in 3 patients, and ureterolithotomy in 2 patients. For the retroperitoneal approach, the Alexis wound retractor with a single port was used. RESULTS: All cases were completed without conversion to standard laparoscopic or open surgery. The mean operative time and mean estimated blood loss for decortication of the renal cysts were 56 minutes (range 45-70 minutes) and 179 mL (range 22-398 mL), respectively; the mean operative time and mean estimated blood loss for extirpative or reconstructive surgery were 156 minutes (range 115-21 minutes) and 403 mL (range 35-1324 mL), respectively. The mean hospital stay was 6.5 days (range 4-14 days). There were no major intraoperative complications, but wound dehiscence and bleeding were noted in two patients. CONCLUSION: Retroperitoneal single-port laparoscopic urologic surgery, particularly in patients requiring free-hand suturing, is technically feasible. The initial clinical experience of organ ablative, extirpative, and reconstructive surgery with this approach is reported. PMID- 19814701 TI - Laser phototherapy as topical prophylaxis against radiation-induced xerostomia. AB - The common consequences of radiotherapy (RT) to the head and neck are oral mucositis, xerostomia, and severe pain. The aim of this study was to verify how laser phototherapy (LPT) used for oral mucositis could influence xerostomia symptoms and hyposalivation of patients undergoing RT. Patients were divided into two groups: 12 individuals receiving three laser irradiations per week (G1) and 10 patients receiving one laser irradiation per week (G2). A diode laser (660 nm, 6 J/cm(2), 0.24 J, 40 mW) was used until completely healing of the lesions or the end of the RT. At the first and last laser sessions, whole resting and stimulated saliva were collected, and questionnaires were administered. According to Wilcoxon and Student statistical test, xerostomia for G1 was lower than for G2 (p < 0.05), and salivary flow rate was no different before and after RT, except for stimulated collection of G2, which was lower (p < 0.05). Our results suggest that LPT can be beneficial as an auxiliary therapy for hypofunction of salivary glands. PMID- 19814702 TI - Effect of 830 nm laser phototherapy on osteoblasts grown in vitro on Biosilicate scaffolds. AB - OBJECTIVE: The purpose of this study was (i) to develop a method for successfully seeding osteoblasts onto a glass-ceramic scaffold designed for use in clinical settings, and (ii) to determine whether the application of laser phototherapy at 830 nm would result in osteoblast proliferation on the glass-ceramic scaffold. BACKGROUND: The use of bioscaffolds is considered a promising strategy for a number of clinical applications where tissue healing is sub-optimal. As in vitro osteoblast growth is a slow process, laser phototherapy could be used to stimulate osteoblast proliferation on bioscaffolds. METHODS: A methodology was developed to seed an osteoblastic (MC3T3) cell line onto a novel glass-ceramic scaffold. Seeded scaffolds were irradiated with a single exposure of 830 nm laser at 10 J/cm(2) (at diode). Non-irradiated seeded scaffolds acted as negative controls. Cell proliferation was assessed seven days after irradiation. RESULTS: Osteoblastic MC3T3 cells were successfully grown on discs composed of a glass ceramic composite. Laser irradiation produced a 13% decrease in MC3T3 cell proliferation on glass-ceramic discs (mean +/- SD = 0.192 +/- 0.002) compared with control (non-irradiated) discs (mean +/-SD = 0.22 +/- 0.002). CONCLUSIONS: Despite successful seeding of bioscaffolds with osteoblasts, laser phototherapy resulted in a reduction in cell growth compared to non-irradiated controls. Future research combining laser phototherapy and glass-ceramic scaffolds should take into account possible interactions of the laser with matrix compounds. PMID- 19814703 TI - Retraction Notice: Clinical Management of Postoperative Vomiting after Strabismus Surgery in Children AB - Formal retraction notice of article entitled Clinical Management of Postoperative Vomiting after Strabismus Surgery in Children (Curr Drug Saf. 2010 Apr; 5(2):132 48) by Dr Y. Fujii. This article is being retracted as a result of:Failure of Dr. Fijii's institution as well as of himself to rationalize the legitimacy of the study and/or its data as stipulated in request by the Editors-in-Chief of the journals extended on April 9, 2012. PMID- 19814714 TI - Efficient expression and secretion of recombinant human growth hormone in the methylotrophic yeast Pichia pastoris: potential applications for other proteins. AB - A simple high yielding process for the production of rhGH (recombinant human growth hormone) in the Pichia pastoris system is described. The approach adopted the addition of surfactants during fermentation along with methanol induction. A Pichia integrant harbouring multiple-copy, non-codon-optimized hGH showed poor expression in complex and defined media. Inclusion of the surfactants Tween 20 or Tween 80 during induction enhanced the expression levels significantly in shake flask studies. The combination of 2 litres of basal salt medium and Tween 20 in a bioreactor culminated in 3 x 10(4)-fold elevated expression of the protein (approximately 500 mg/l) as estimated by ELISA. SDS/PAGE and Western-blot analyses revealed that the Pichia-derived rhGH migrated as a single band with a molecular mass of approximately 22 kDa and had the same immunoreactivity as native commercial rhGH. Analysis of Pichia-derived purified rhGH and commercial rhGH on an Agilent 2100 Bioanalyzer revealed overlapping peaks displaying authentic N-terminal processing of Pichia-derived rhGH, which was further confirmed by N-terminal sequencing. In addition, matrix-assisted laser-desorption ionization-time of flight analysis of the protein confirmed its authenticity. These results indicate that the P. pastoris expression system can be effective in the production of rhGH at commercially relevant levels. PMID- 19814715 TI - Role of the cysteine protease interpain A of Prevotella intermedia in breakdown and release of haem from haemoglobin. AB - The gram-negative oral anaerobe Prevotella intermedia forms an iron(III) protoporphyrin IX pigment from haemoglobin. The bacterium expresses a 90 kDa cysteine protease, InpA (interpain A), a homologue of Streptococcus pyogenes streptopain (SpeB). The role of InpA in haemoglobin breakdown and haem release was investigated. At pH 7.5, InpA mediated oxidation of oxyhaemoglobin to hydroxymethaemoglobin [in which the haem iron is oxidized to the Fe(III) state and which carries OH- as the sixth co-ordinate ligand] by limited proteolysis of globin chains as indicated by SDS/PAGE and MALDI (matrix-assisted laser desorption ionization)-TOF (time-of-flight) analysis. Prolonged incubation at pH 7.5 did not result in further haemoglobin protein breakdown, but in the formation of a haemoglobin haemichrome (where the haem Fe atom is co-ordinated by another amino acid ligand in addition to the proximal histidine residue) resistant to degradation by InpA. InpA-mediated haem release from hydroxymethaemoglobin agarose was minimal compared with trypsin at pH 7.5. At pH 6.0, InpA increased oxidation at a rate greater than auto-oxidation, producing aquomethaemoglobin (with water as sixth co-ordinate ligand), and resulted in its complete breakdown and haem loss. Aquomethaemoglobin proteolysis and haem release was prevented by blocking haem dissociation by ligation with azide, whereas InpA proteolysis of haem-free globin was rapid, even at pH 7.5. Both oxidation of oxyhaemoglobin and breakdown of methaemoglobin by InpA were inhibited by the cysteine protease inhibitor E-64 [trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane]. In summary, we conclude that InpA may play a central role in haem acquisition by mediating oxyhaemoglobin oxidation, and by degrading aquomethaemoglobin in which haem-globin affinity is weakened under acidic conditions. PMID- 19814723 TI - The end of a myth: cloning and characterization of the ovine melatonin MT(2) receptor. AB - BACKGROUND AND PURPOSE: For many years, it was suspected that sheep expressed only one melatonin receptor (closely resembling MT(1) from other mammal species). Here we report the cloning of another melatonin receptor, MT(2), from sheep. EXPERIMENTAL APPROACH: Using a thermo-resistant reverse transcriptase and polymerase chain reaction primer set homologous to the bovine MT(2) mRNA sequence, we have cloned and characterized MT(2) receptors from sheep retina. KEY RESULTS: The ovine MT(2) receptor presents 96%, 72% and 67% identity with cattle, human and rat respectively. This MT(2) receptor stably expressed in CHO-K1 cells showed high-affinity 2[(125)I]-iodomelatonin binding (K(D)= 0.04 nM). The rank order of inhibition of 2[(125)I]-iodomelatonin binding by melatonin, 4-phenyl-2 propionamidotetralin and luzindole was similar to that exhibited by MT(2) receptors of other species (melatonin > 4-phenyl-2-propionamidotetralin > luzindole). However, its pharmacological profile was closer to that of rat, rather than human MT(2) receptors. Functionally, the ovine MT(2) receptors were coupled to G(i) proteins leading to inhibition of adenylyl cyclase, as the other melatonin receptors. In sheep brain, MT(2) mRNA was expressed in pars tuberalis, choroid plexus and retina, and moderately in mammillary bodies. Real-time polymerase chain reaction showed that in sheep pars tuberalis, premammillary hypothalamus and mammillary bodies, the temporal pattern of expression of MT(1) and MT(2) mRNA was not parallel in the three tissues. CONCLUSION AND IMPLICATIONS: Co-expression of MT(1) and MT(2) receptors in all analysed sheep brain tissues suggests that MT(2) receptors may participate in melatonin regulation of seasonal anovulatory activity in ewes by modulating MT(1) receptor action. PMID- 19814724 TI - Protective role of the antidiabetic drug metformin against chronic experimental pulmonary hypertension. AB - BACKGROUND AND PURPOSE: Pulmonary arterial hypertension (PAH) is associated with increased contraction and proliferation of pulmonary vascular smooth muscle cells. The anti-diabetic drug metformin has been shown to have relaxant and anti proliferation properties. We thus examined the effect of metformin in PAH. EXPERIMENTAL APPROACH: Metformin effects were analysed in hypoxia- and monocrotaline-induced PAH in rats. Ex vivo and in vitro analyses were performed in lungs, pulmonary artery rings and cells. KEY RESULTS: In hypoxia- and monocrotaline-induced PAH, the changes in mean pulmonary arterial pressure and right heart hypertrophy were nearly normalized by metformin treatment (100 mg.kg( 1).day(-1)). Pulmonary arterial remodelling occurring in both experimental models of PAH was also inhibited by metformin treatment. In rats with monocrotaline induced PAH, treatment with metformin significantly increased survival. Metformin increased endothelial nitric oxide synthase phosphorylation and decreased Rho kinase activity in pulmonary artery from rats with PAH. These effects are associated with an improvement of carbachol-induced relaxation and reduction of phenylephrine-induced contraction of pulmonary artery. In addition, metformin inhibited mitogen-activated protein kinase activation and strongly reduced pulmonary arterial cell proliferation during PAH. In vitro, metformin directly inhibited pulmonary artery smooth muscle cell growth. CONCLUSIONS AND IMPLICATIONS: Metformin protected against PAH, regardless of the initiating stimulus. This protective effect may be related to its anti-remodelling property involving improvement of endothelial function, vasodilatory and anti proliferative actions. As metformin is currently prescribed to treat diabetic patients, assessment of its use as a therapy against PAH in humans should be easier. PMID- 19814725 TI - Long-term effects of losartan on structure and function of the thoracic aorta in a mouse model of Marfan syndrome. AB - BACKGROUND AND PURPOSE: During development of thoracic aortic aneurysms in a mouse model of Marfan syndrome, upregulation of matrix metalloproteinase (MMP)-2 and -9 was accompanied by compromised aortic constriction and endothelium dependent relaxation. Losartan has been proposed for the prevention of thoracic aortic aneurysm. We hypothesized that losartan would suppress MMP-2/-9 activation and improve aortic vasomotor function in this model. EXPERIMENTAL APPROACH: A well-characterized mouse model of Marfan syndrome (Fbn1(C1039G/+)) was used. Starting at 6 weeks old, Marfan mice were untreated or given losartan (0.6 g.L( 1) in drinking water, n= 30). The littermate Fbn1(+/+) mice served as control. Thoracic aortas were studied at 3, 6 and 9 months by histology and by contractility assays in isolated segments in vitro. KEY RESULTS: Losartan improved elastic fibre organization and increased aortic breaking stress. Losartan reduced the activity and protein expression of MMP-2 and MMP-9 at all ages. Aortic constriction in response to membrane depolarization or phenylephrine was increased by losartan at 3 and 9 months by 100-200%. Active force of aortic smooth muscle was also increased at 6 and 9 months. Acetylcholine-induced endothelium-dependent relaxation was improved by 30% after 3 months of losartan treatment, but such improvement disappeared with longer duration of treatment, accompanied by reduced phosphorylation of endothelial nitric oxide (NO) synthase(Ser1177), Akt(Thr308) and Akt(Ser473), compared with the control. CONCLUSIONS AND IMPLICATIONS: Losartan improved the contractile function of aorta and reduced MMP activation. However, the endothelial NO pathway remained suppressed in the thoracic aorta during losartan treatment, which might limit its long-term benefits in Marfan syndrome. PMID- 19814726 TI - Dysfunction of endothelial and smooth muscle cells in small arteries of a mouse model of Marfan syndrome. AB - BACKGROUND AND PURPOSE: Marfan syndrome, a connective tissue disorder caused by mutations in FBN1 encoding fibrillin-1, results in life-threatening complications in the aorta, but little is known about its effects in resistance vasculature. EXPERIMENTAL APPROACH: Second-order mesenteric arteries from mice at 3, 6 and 10 months of age (n= 30) heterozygous for the Fbn1 allele encoding a cysteine substitution (Fbn1(C1039G/+)) were compared with those from age-matched control littermates. KEY RESULTS: Stress-strain curves indicated that arterial stiffness was increased at 6 and 10 months of age in Marfan vessels. Isometric force measurement revealed that contraction in response to potassium (60 mM)-induced membrane depolarization was decreased by at least 28% in Marfan vessels at all ages, while phenylephrine (3 microM)-induced contraction was reduced by at least 40% from 6 months. Acetylcholine-induced relaxation in Marfan vessels was reduced to 70% and 45% of control values, respectively, at 6 and 10 months. Sensitivity to sodium nitroprusside was reduced at 6 months (pEC(50)= 5.64 +/- 0.11, control pEC(50)= 7.34 +/- 0.04) and 10 months (pEC(50)= 5.99 +/- 0.07, control pEC(50)= 6.99 +/- 0.14). Pretreatment with N(omega)-Nitro-L-arginine methyl ester (200 microM) had no effect on acetylcholine-induced relaxation in Marfan vessels, but reduced vasorelaxation in control vessels to 57% of control values. Addition of indomethacin (10 microM) and catalase (1000 U.mL(-1)) further inhibited vasorelaxation in Marfan vessels to a greater degree compared with control vessels. CONCLUSIONS AND IMPLICATIONS: Pathogenesis of Marfan syndrome in resistance-sized arteries increases stiffness and impairs vasomotor function. PMID- 19814727 TI - Cloning and pharmacological characterization of the dog P2X7 receptor. AB - BACKGROUND AND PURPOSE: Human and rodent P2X7 receptors exhibit differences in their sensitivity to antagonists. In this study we have cloned and characterized the dog P2X7 receptor to determine if its antagonist sensitivity more closely resembles the human or rodent orthologues. EXPERIMENTAL APPROACH: A cDNA encoding the dog P2X7 receptor was isolated from a dog heart cDNA library, expressed in U 2 OS cells using the BacMam viral expression system and characterized in electrophysiological, ethidium accumulation and radioligand binding studies. Native P2X7 receptors were examined by measuring ATP-stimulated interleukin-1beta release in dog and human whole blood. KEY RESULTS: The dog P2X7 receptor was 595 amino acids long and exhibited high homology (>70%) to the human and rodent orthologues although it contained an additional threonine at position 284 and an amino acid deletion at position 538. ATP possessed low millimolar potency at dog P2X7 receptors. 2'-&3'-O-(4benzoylbenzoyl) ATP had slightly higher potency but was a partial agonist. Dog P2X7 receptors possessed relatively high affinity for a number of selective antagonists of the human P2X7 receptor although there were some differences in potency between the species. Compound affinities in human and dog blood exhibited a similar rank order of potency as observed in studies on the recombinant receptor although absolute potency was considerably lower. CONCLUSIONS AND IMPLICATIONS: Dog recombinant and native P2X7 receptors display a number of pharmacological similarities to the human P2X7 receptor. Thus, dog may be a suitable species for assessing target-related toxicity of antagonists intended for evaluation in the clinic. PMID- 19814728 TI - Irbesartan-mediated reduction of renal and cardiac damage in insulin resistant JCR : LA-cp rats. AB - BACKGROUND AND PURPOSE: Angiotensin II receptor antagonists (ARBs), originally developed for antihypertensive properties, have pleiotropic effects including direct vascular actions. We tested the hypothesis that the ARB irbesartan would be effective against micro- and macrovascular complications of the prediabetic metabolic syndrome using the obese, insulin-resistant JCR : LA-cp rat that exhibits micro- and macrovascular disease with ischaemic myocardial lesions and renal disease. EXPERIMENTAL APPROACH: Obese male rats were treated with irbesartan (30 mg.kg(-1).day(-1), incorporated into chow) from 12 to 25 weeks of age. KEY RESULTS: Irbesartan treatment caused no change in food intake or body weight. Fasting glycaemic control of the JCR : LA-cp rats was marginally improved, at the expense of increased plasma insulin levels ( approximately 50%). Fasting plasma triglycerides were marginally reduced ( approximately 25%), while cholesterol concentrations were unchanged. Elevated concentrations of adiponectin, monocyte chemotactic protein-1 and plasminogen activator inhibitor-1 were reduced along with severity of glomerular sclerosis. Macrovascular dysfunction (aortic hypercontractile response to noradrenergic stimulus and reduced endothelium-dependent relaxation) was improved and frequency of ischaemic myocardial lesions reduced (62%). CONCLUSIONS AND IMPLICATIONS: Irbesartan reduces markers of inflammation and prothombotic status, improves macrovascular function and reduces glomerular sclerosis and myocardial lesions in a model of the metabolic syndrome. Unlike pharmaceutical agents targeted on metabolic dysfunction, irbesartan reduced end-stage disease without major reduction of plasma lipids or insulin. The protective effects appear to be secondary to unknown intracellular mechanisms, probably involving signal transduction pathways. Understanding these would offer novel pharmaceutical approaches to protection against cardiovascular disease. PMID- 19814729 TI - FTY720 (fingolimod) in Multiple Sclerosis: therapeutic effects in the immune and the central nervous system. AB - FTY720 (fingolimod) is a first-in-class sphingosine 1-phosphate (S1P) receptor modulator that was highly effective in Phase II clinical trials for Multiple Sclerosis (MS). FTY720 is phosphorylated in vivo by sphingosine kinase-2 to form the active moiety FTY720-phosphate that binds to four of the five G protein coupled S1P receptor subtypes. Studies using conditional S1P1 receptor-deficient and sphingosine kinase-deficient mice showed that the egress of lymphocytes from lymph nodes requires signalling of lymphocytic S1P1 receptors by the endogenous ligand S1P. The S1P mimetic FTY720-phosphate causes internalization and degradation of cell membrane-expressed S1P1, thereby antagonizing S1P action at the receptor. In models of human MS and demyelinating polyneuropathies, functional antagonism of lymphocytic S1P1 slows S1P-driven egress of lymphocytes from lymph nodes, thereby reducing the numbers of autoaggressive TH17 cells that recirculate via lymph and blood to the central nervous system and the sciatic/ischiatic nerves. Based on its lipophilic nature, FTY720 crosses the blood-brain barrier, and ongoing experiments suggest that the drug also down modulates S1P1 in neural cells/astrocytes to reduce astrogliosis, a phenomenon associated with neurodegeneration in MS. This may help restore gap-junctional communication of astrocytes with neurons and cells of the blood-brain barrier. Additional effects may result from (down-) modulation of S1P3 in astrocytes and of S1P1 and S1P5 in oligodendrocytes. In conclusion, FTY720 may act through immune-based and central mechanisms to reduce inflammation and support structural restoration of the central nervous system parenchyma. Beyond the autoimmune indications, very recent studies suggest that short-term, low-dose administration of FTY720 could help treat chronic (viral) infections. Differential effects of the drug on the trafficking of naive, central memory and effector memory T cell subsets are discussed. PMID- 19814730 TI - Tobacco smoke affects expression of peroxisome proliferator-activated receptor gamma in monocyte/macrophages of patients with coronary heart disease. AB - BACKGROUND AND PURPOSE: Tobacco smoke represents a relevant risk factor for coronary heart disease (CHD). Although peroxisome proliferator-activated receptor (PPAR)gamma activation reduces inflammation and atherosclerosis, expression of PPARgamma in cells and its modulation by smoking are poorly investigated. We previously reported that monocyte/macrophages from healthy smokers exhibited an enhanced constitutive expression of PPARgamma. Here, we evaluated PPARgamma expression and basal cytokine release in monocytes and monocyte-derived macrophages (MDMs) from 85 CHD patients, classified by their smoking habit (smokers, non-smokers and ex-smokers), and assessed the role of PPARgamma ligands in this context. EXPERIMENTAL APPROACH: PPARgamma protein was detected by Western blot and semi-quantified by PPARgamma/beta-actin ratio; cytokine release was measured by elisa and nuclear factor-kappaB (NF-kappaB) translocation by electrophoretic mobility shift assays. KEY RESULTS: As compared to the other groups, MDMs from smoker CHD patients exhibited a reduced PPARgamma/beta-actin ratio and an increased spontaneous release of tumour necrosis factor-alpha (TNF alpha) and interleukin-6, but with no major variations in monocytes. In cells from selected CHD patients, rosiglitazone inhibited TNF-alpha release and NF kappaB translocation induced by phorbol-12-myristate 13-acetate. The selective PPARgamma antagonist GW9662 reversed these effects, with some variations related to smoking habit. CONCLUSIONS AND IMPLICATIONS: In CHD patients, exposure to tobacco smoke profoundly affected PPARgamma expression, and this was related to levels of secretion of pro-inflammatory cytokines. MDMs from CHD smokers showed the lowest PPARgamma expression and released more inflammatory cytokines. Moreover, rosiglitazone's ability to inhibit cytokine release and its reversal by GW9662 clearly indicated PPARgamma involvement in these changes in CHD patients. PMID- 19814731 TI - Differential effects of quercetin, apigenin and genistein on signalling pathways of protease-activated receptors PAR(1) and PAR(4) in platelets. AB - BACKGROUND AND PURPOSE: The modulation by flavonoids of platelet responses induced by thrombin has been little investigated, and the antiplatelet activity, as well as possible inhibitory mechanisms of these compounds on thrombin signalling, has not yet been elucidated. We explored whether flavonoids affect platelet signalling pathways triggered by thrombin and by the selective activation of its protease-activated receptors (PARs) 1 and 4, and analysed the antagonism of these polyphenols at thrombin receptors. EXPERIMENTAL APPROACH: We investigated the effect of a range of polyphenolic compounds on platelet aggregation, 5-HT secretion, intracellular calcium mobilization, protein kinase activity and tyrosine phosphorylation, triggered by thrombin and PAR agonist peptides (PAR-APs). The ability of these flavonoids to bind to thrombin receptors was investigated by competitive radioligand binding assays using (125)I-thrombin. KEY RESULTS: Quercetin, apigenin and genistein impaired platelet aggregation, as well as 5-HT release and calcium mobilization, induced by thrombin and PAR-APs. Quercetin and apigenin were inhibitors of protein kinases, but genistein exhibited a minimal ability to suppress platelet phosphorylation. Binding assays did not establish any kind of interaction between thrombin receptors and any of the flavonoids tested. CONCLUSIONS AND IMPLICATIONS: Quercetin, apigenin and genistein did not inhibit thrombin responses by interacting with thrombin receptors, but by interfering with intracellular signalling. While inhibition by genistein may be a consequence of affecting calcium mobilization, subsequent platelet secretion and aggregation, for quercetin and apigenin, inhibition of kinase activation may also be involved in the impairment of platelet responses. PMID- 19814732 TI - Vitamin D inhibits growth of human airway smooth muscle cells through growth factor-induced phosphorylation of retinoblastoma protein and checkpoint kinase 1. AB - BACKGROUND AND PURPOSE: Airway remodelling in asthma is manifested, in part, as increased airway smooth muscle (ASM) mass, reflecting myocyte proliferation. We hypothesized that calcitriol, a secosteroidal vitamin D receptor (VDR) modulator, would inhibit growth factor-induced myocyte proliferation. EXPERIMENTAL APPROACH: Human ASM cell cultures were derived from bronchial samples taken during surgery. ASM cells were treated with platelet-derived growth factor (PDGF) (10 ng.mL(-1)) for 24 h in the presence of calcitriol, dexamethasone or a checkpoint kinase 1 (Chk1) inhibitor (SB218078). The effects of calcitriol on PDGF-mediated cell proliferation were assessed by thymidine incorporation assay, propidium iodide based cell cycle analysis, caspase-3 assay and immunoblotting for specific cell cycle modulators. KEY RESULTS: Calcitriol, but not dexamethasone, inhibited PDGF induced ASM DNA synthesis concentration dependently (IC(50)= 520 +/- 52 nM). These effects were associated with VDR-mediated expression of cytochrome CYP24A1 with no effects on ASM apoptosis. Calcitriol substantially inhibited (P < 0.01) PDGF-stimulated cell growth in ASM derived from both normal (59 +/- 8%) and asthmatic subjects (57 +/- 9%). Calcitriol inhibited PDGF-induced phosphorylation of retinoblastoma protein (Rb) and Chk1, with no effects on PDGF-mediated activation of extracellular signal-regulated kinases 1/2, PI3-kinase and S6 kinase, or expression of p21(Waf/Cip-1), p27(Kip1), cyclin D and E2F-1. Consistent with these observations, SB218078 also inhibited (IC(50)= 450 +/- 100 pM) PDGF-induced cell cycle progression. CONCLUSIONS AND IMPLICATIONS: Calcitriol decreased PDGF-induced ASM cell growth by inhibiting Rb and Chk1 phosphorylation. PMID- 19814734 TI - PC-407, a celecoxib derivative, inhibited the growth of colorectal tumor in vitro and in vivo. AB - This study aimed to observe the growth-inhibitory effect of PC-407 (4-[5-naphthyl 3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide), a celecoxib derivative synthesized in our lab, in human colorectal cancer cells and a colitis-associated colorectal cancer (CACC) model, and investigate the relative molecular mechanisms. SW-1116 (expressing a high level of cyclooxygenase-2 [COX-2]), HT-29 (expressing a moderate level of COX-2), and SW-480 (no expression of COX-2) cell lines were exposed to different concentrations of celecoxib (0-100 micromol/L) or PC-407 (0-100 micromol/L). Then, COX-2 levels were assessed by reverse transcription-PCR and Western blotting. COX-2 activity was evaluated by measuring prostaglandin E(2) concentration using enzyme-linked immunoassay. A mouse model of colitis-associated carcinogenesis was employed to determine the effect of PC 407 in vivo. PC-407 inhibited cell growth in a concentration-dependent manner, and the IC(50) values of PC-407 for growth inhibition of SW-1116, HT-29, and SW 480 cells were 17.60 +/- 3.02, 18.14 +/- 2.81, and 8.13 +/- 0.40 micromol/L, respectively. PC-407 down-regulated COX-2 mRNA and protein levels and reduced prostaglandin E(2) production significantly. In vivo, PC-407 inhibited the genesis of CACC effectively. Our data indicate that PC-407 can inhibit the growth of tumor both in vitro and in vivo and suggest that the effect probably involves inhibition of the COX-2 pathway and other COX-2-independent pathways. PMID- 19814733 TI - Visualization of gene expression in the live subject using the Na/I symporter as a reporter gene: applications in biotherapy. AB - Biotherapies involve the utilization of antibodies, genetically modified viruses, bacteria or cells for therapeutic purposes. Molecular imaging has the potential to provide unique information that will guarantee their biosafety in humans and provide a rationale for the future development of new generations of reagents. In this context, non-invasive imaging of gene expression is an attractive prospect, allowing precise, spacio-temporal measurements of gene expression in longitudinal studies involving gene transfer vectors. With the emergence of cell therapies in regenerative medicine, it is also possible to track cells injected into subjects. In this context, the Na/I symporter (NIS) has been used in preclinical studies. Associated with a relevant radiotracer ((123)I(-), (124)I(-), (99m)TcO4(-)), NIS can be used to monitor gene transfer and the spread of selectively replicative viruses in tumours as well as in cells with a therapeutic potential. In addition to its imaging potential, NIS can be used as a therapeutic transgene through its ability to concentrate therapeutic doses of radionuclides in target cells. This dual property has applications in cancer treatment and could also be used to eradicate cells with therapeutic potential in the case of adverse events. Through experience acquired in preclinical studies, we can expect that non-invasive molecular imaging using NIS as a transgene will be pivotal for monitoring in vivo the exact distribution and pharmacodynamics of gene expression in a precise and quantitative way. This review highlights the applications of NIS in biotherapy, with a particular emphasis on image-guided radiotherapy, monitoring of gene and vector biodistribution and trafficking of stem cells. PMID- 19814736 TI - Clinical and laboratory phenotype associated with the aspirin-like defect. PMID- 19814738 TI - Interferon as an alternative to purine analogues in the treatment of hairy cell leukaemia. PMID- 19814739 TI - Advances in the understanding of acquired haemophilia A: implications for clinical practice. AB - Acquired haemophilia A is an auto-immune disease caused by an inhibitory antibody to factor VIII. Patients with an acquired factor VIII inhibitor are at risk of life- and limb-threatening bleeding until the inhibitor has been eradicated. Management relies on rapid and accurate diagnosis, control of bleeding episodes, investigation for a precipitating cause and eradication of the inhibitor by immunosuppression. Patients should always be managed jointly with a specialist centre even if they present without overt bleeding. Despite an extensive literature, few controlled data are available and management guidelines are predominantly based on case reports, retrospective cohorts and expert opinion. This paper reviews the current literature on incidence, pathogenesis, diagnosis, haemostatic therapy and inhibitor eradication strategies. Potential future developments are discussed. PMID- 19814740 TI - The fundamental contribution of William Bate Hardy to shape the concept of mast cell heterogeneity. AB - This review article acknowledges the pioneering contribution of William Bate Hardy in shaping the concept of mast cell heterogeneity. In two outstanding papers, published in 1894 and 1895, he focussed on the 'wandering cells' (the modern leucocytes) in different mammalian species and distinguished two types of granular basophil cells, i.e., the coarsely granular basophil cells and the splanchnic basophil cells. These corresponded to the populations of connective tissue-type and mucosal mast cells, respectively, described 70 years later by Enerback in rodents. Among the coarsely granular basophil cells, he also differentiated those cells which populated the serosal cavities - the so-called coelomic coarsely granular basophil cells - from the common coarsely granular basophil cells, which were localized in the connective tissues. He stated that the granular basophil cells presented with different morphological and histochemical characteristics in diverse animal species as well as at different anatomical sites. Remarkably, he performed a series of functional experiments on the basophil cells as well as the other wandering cells, and suggested the view that different granular basophil cells might express functional specializations. PMID- 19814741 TI - Circulating mesothelial cells following multiple ribs fractures. PMID- 19814742 TI - Laboratory monitoring of Scott Syndrome. PMID- 19814743 TI - The assessment of regional gut transit times in healthy controls and patients with gastroparesis using wireless motility technology. AB - BACKGROUND: Wireless pH and pressure motility capsule (wireless motility capsule) technology provides a method to assess regional gastrointestinal transit times. AIMS: To analyse data from a multi-centre study of gastroparetic patients and healthy controls and to compare regional transit times measured by wireless motility capsule in healthy controls and gastroparetics (GP). METHODS: A total of 66 healthy controls and 34 patients with GP (15 diabetic and 19 idiopathic) swallowed wireless motility capsule together with standardized meal (255 kcal). Gastric emptying time (GET), small bowel transit time (SBTT), colon transit time (CTT) and whole gut transit time (WGTT) were calculated using the wireless motility capsule. RESULTS: Gastric emptying time, CTT and WGTT but not SBTT were significantly longer in GP than in controls. Eighteen percent of gastroparetic patients had delayed WGTT. Both diabetic and idiopathic aetiologies of gastroparetics had significantly slower WGTT (P < 0.0001) in addition to significantly slower GET than healthy controls. Diabetic gastroparetics additionally had significantly slower CTT than healthy controls (P = 0.0054). CONCLUSIONS: In addition to assessing gastric emptying, regional transit times can be measured using wireless motility capsule. The prolongation of CTT in gastroparetic patients indicates that dysmotility beyond the stomach in GP is present, and it could be contributing to symptom presentation. PMID- 19814744 TI - Review article: influenza A (H1N1) virus in patients with inflammatory bowel disease. AB - BACKGROUND: Infection with influenza A (H1N1)v (swine flu) has caused widespread anxiety, among patients who are potentially immunocompromised, such as those being treated for inflammatory bowel disease. AIM: To provide guidance for physicians and their patients on the risk, prevention and management of influenza A (H1N1)v infection. METHODS: Medline was searched using the following key words: 'swine flu', 'immunosuppression', inflammatory bowel disease', 'recommendations', 'immunization', 'vaccination'. Organizations such as European Centre for Disease Prevention and Control, the Centers for Disease Control and Prevention and the World Health Organization were consulted for recent papers and recommendations regarding immunocompromised patients and influenza A (H1N1)v infection. RESULTS: Pandemic influenza A (H1N1) virus predominantly affects young patients. Those who are immunocompromised because of underlying disease or treatment are considered at higher risk of complications from influenza A (H1N1). They should be offered prevention (vaccination, postexposure prophylaxis) or treatment with antiviral drugs, if affected. Pneumococcal infection is a complication of influenza infection; therefore, pneumococcal vaccination appears advisable. Seasonal influenza vaccination is also recommended. Withdrawal of immunosuppressive treatment appears advisable during severe active infection if possible. CONCLUSIONS: Pragmatic advice is the best that can be offered in the current circumstances because of paucity of evidence. Investigation into the impact of influenza A (H1N1)v infection in young people with chronic conditions is needed. PMID- 19814745 TI - Systematic review: distribution of advanced neoplasia according to polyp size at screening colonoscopy. AB - BACKGROUND: The impact of not referring sub-centimetre polyps identified at CT colonography upon the efficacy of colorectal cancer screening remains uncertain. AIM: To determine the distribution of advanced neoplasia according to polyp size in a screening setting. METHODS: Published studies reporting the distribution of advanced adenomas in asymptomatic screening cohorts according to polyp size were identified by MEDLINE and EMBASE searches. Predefined outputs were the screening rates of advanced adenomas represented by diminutive (< or =5 mm), small (6-9 mm), sub-centimetre (<10 mm) and large (> or =10 mm) polyp sizes. RESULTS: Data from four studies with 20 562 screening subjects met the primary inclusion criteria. Advanced adenomas were detected in 1155 (5.6%) subjects (95% CI = 5.3 5.9), corresponding to diminutive, small and large polyps in 4.6% (95% CI = 3.4 5.8), 7.9% (95% CI = 6.3-9.4) and 87.5% (95% CI = 86-89.4) of cases respectively. The frequency of advanced lesions among patients whose largest polyp was diminutive, small, sub-centimetre and large in size was 0.9%, 4.9%, 1.7% and 73.5% respectively. CONCLUSIONS: Based on this systematic review, a 6-mm polyp size threshold for polypectomy referral would identify over 95% of subjects with advanced adenomas, whereas a 10-mm threshold would identify 88% of cases. PMID- 19814746 TI - Irritable bowel syndrome symptoms and health related quality of life in female veterans. AB - BACKGROUND: The status and determinants of health-related quality of life (HRQOL) in female veterans with and without irritable bowel syndrome (IBS) are unknown. AIM: To compare HRQOL in female veterans with and without IBS symptoms and examine the contribution of post-traumatic stress disorder, depression and anxiety to HRQOL. METHODS: A cross-sectional study of 339 female veterans. Self report questionnaires were used to evaluate IBS symptoms, post-traumatic stress disorder, depression, anxiety and HRQOL. RESULTS: Symptoms consistent with IBS were present in 33.5% of participants. Female veterans with IBS symptoms had significant reductions in physical component score and 5 of 8 Health Related Quality of Life subscales and on 7 of 8 Irritable Bowel Syndrome Quality Of Life subscales than female veterans without IBS symptoms. Compared with the US general female population, female veterans had significantly lower Health Related Quality of Life physical component score and mental component scores (MCS) irrespective of IBS symptom status. Differences in the MCS score were most explained by depression, while those in the physical component score were most explained by anxiety. CONCLUSIONS: Irritable bowel syndrome symptoms in female veterans are associated with considerable reduction in HRQOL. However, female veterans, regardless of IBS symptom status, have lower HRQOL compared with the general US female population. PMID- 19814747 TI - Screening for hepatitis B in chemotherapy patients: survey of current oncology practices. AB - BACKGROUND: Hepatitis B virus (HBV) reactivation occurs in up to 78% of patients receiving cytotoxic chemotherapy for nonhepatic malignancies. Reactivation can lead to hepatic dysfunction, jaundice and fulminant hepatic failure. Current recommendations include screening patients at risk for HBV prior to immunosuppressive therapy and initiating antiviral prophylaxis in patients with chronic HBV. AIM: To investigate current practice among oncologists regarding HBV screening and antiviral prophylaxis in candidates for chemotherapy. METHODS: A survey was sent to American Medical Association registered oncologists assessing demographics and HBV screening practices. Statistical analysis was performed using Fisher's exact test. RESULTS: In all, 265 responses were received. Office based physicians were less likely to screen for HBV prior to chemotherapy (P < 0.001). Years in practice varied: 51% with <5 years, 29% with 5-15 years and 18% with >15 years, with no difference in screening practices between groups (P = N.S.). Responders screen for HBV as follows: never - 20%, only in the presence of abnormal liver biochemistries - 30%, risk factors or history of hepatitis - 38%. In patients with known HBV, 75% of oncologists refer to specialists, 7% initiate therapy, while 15% do not refer or initiate therapy, most of whom are in an office setting (P = 0.02). CONCLUSIONS: Twenty per cent of oncologists never screen for HBV prior to initiating chemotherapy. Office-based physicians were less likely to screen, treat or refer to a specialist prior to chemotherapy. Greater education regarding risk of HBV reactivation is needed for clinicians treating patients with immunosuppressive therapies. PMID- 19814748 TI - A comparison of a flexometallic tracheal tube with the intubating laryngeal mask tracheal tube for nasotracheal fibreoptic intubation using the two-scope technique. AB - We compared the incidence and site of impingement of a flexometallic tracheal tube with those of the re-usable intubating laryngeal mask (ILMA) tube in 60 anaesthetised patients undergoing nasotracheal fibreoptic intubation for oral surgery. A two-scope technique was used, observing the site of impingement with one scope whilst intubating with the other. The tubes were 6.0-mm in females and 6.5-mm in males. Impingement occurred with 10 (33%) flexometallic and 2 (7%) ILMA tubes (p < 0.032). In all but one case, the impingement was posterior to the right arytenoid cartilage. When impingement was observed, a single disempaction with a 90 degrees anticlockwise rotational manoeuvre overcame impingement in every case except one, allowing successful intubation. We conclude that the incidence of impingement of the tracheal tube, and therefore of potential laryngeal trauma from nasotracheal fibreoptic intubation, is significantly greater with the flexometallic tube than with the ILMA tube. PMID- 19814749 TI - Gestational diabetes and offspring body disproportion. AB - AIM: It has been demonstrated that females born large for gestational age (LGA) in weight but not length are at increased risk of being obese at childbearing age. We addressed the question whether women with gestational diabetes mellitus (GDM) are at increased risk of giving birth to such infants. METHODS: Birth characteristics of 884,267 infants of non-diabetic mothers and 7817 of mothers with GDM were analysed. LGA was defined as birth weight or birth length >2 standard deviation scores for gestational age. Multiple logistic regression analysis was performed. RESULTS: The odds ratio (OR) for a woman with GDM to give birth to an LGA infant that was heavy alone was four times increased (OR: 3.71, 95% CI: 3.41-4.04). Furthermore, in the population of mothers giving birth to LGA infants, the proportion heavy alone was 68% in the group of women with GDM compared with 64.4% in the group of non-diabetic women. The risks were independent of gender of the foetus. CONCLUSION: Women with GDM have an almost four times higher risk of delivering an LGA infant that is heavy alone. The noted disproportion between weight and length in infants of such mothers may have an impact on the risk of later obesity. PMID- 19814750 TI - The definition of human viability: a historical perspective. AB - AIM: To investigate the current concepts related to the definition of human viability in a historical perspective. METHODS: A review of the international literature was performed by using PubMed, Google, Embase to find definitions of Human Viability. The papers collected were divided chronologically. RESULTS: Over the years, many authors have debated the concept of the limit of human viability, starting from Ballantyne in the early 1900. The definition differs from author to author. CONCLUSION: The definition of human viability is still under discussion. It depends primarily, but not only, on gestational age. A number of genetic, cultural, social and technological variables are also influential. This definition could have an impact on legislation and the widespread misunderstanding of this concept in the lay public could also be enhanced by a frank discussion among clinicians and other professionals. PMID- 19814751 TI - Asylum-seeking children with severe loss of activities of daily living: clinical signs and course during rehabilitation. AB - AIM: To investigate whether severe loss of activities of daily living (ADL) in asylum-seeking children is associated with physical disease or toxic influences and to describe the clinical course during the recovery process. METHODS: A total of 29 asylum-seeking children with severe loss of ADL were regularly assessed by physical examinations, laboratory tests and a structured evaluation of their ADL status during rehabilitation. RESULTS: A total of 12 children had previously recorded suicide attempts and 21 were recorded to have experienced traumatic events in their country of origin. The mean time from turning point to recovery was 6 months. Of the study participants, 22 needed enteral feeding and 18 gained weight during recovery. All children had a pulse rate and systolic blood pressure within the normal range. No sign of intoxication or physical disease was identified in laboratory tests or clinical examinations, with the exception of one case of epilepsy. CONCLUSION: Physical disease, pharmacological sedation or anorexia nervosa was not considered to be a probable cause of the loss of ADL in these children. The high rate of psychosocial risk factors and the stressful event of being in an asylum-seeking process call for further investigation of psychosomatic mechanisms. PMID- 19814752 TI - Mastoid subperiosteal abscess as a first sign of unnoticed cholesteatoma in children. AB - AIM: To present children who underwent mastoidectomy for congenital cholesteatoma presented as a subperiosteal abscess. RESULTS: All seven children (age range 7-14 years, six boys) presented with retroauricular swelling, erythema and fluctuation in the mastoid area, and one child also had a mastoid-cutaneous fistula. Five children had otorrhoea, while the other two had normal-appearing tympanic membranes. None of the children had a history of middle ear disease. Four children were treated with antibiotics for a recent episode of otitis media prior to admission. The main findings at surgery were pus, granulations and erosion of the mastoid cortex. Pseudomonas aeruginosa and Proteus sp. were isolated from the abscess in two patients, and the other five cultures were negative. All the patients demonstrated some degree of hearing impairment after surgery. CONCLUSION: Surgical eradication of a mastoid SA in older children is essential as it may be the first indication of an underlying CC. PMID- 19814753 TI - Low frequency of VHL germline mutations in Norwegian patients presenting with isolated central nervous system hemangioblastomas--a population-based study. AB - OBJECTIVES: Explore the genetic and clinical incidence of von Hippel-Lindau disease in patients presenting with isolated central nervous system hemangioblastomas. RESULTS: We report a 3.2% (1/31) and 25% (8/32) incidence of genetic and clinical VHL, respectively. One patient tested positive for a VHL mutation that has not previously been reported. This genotype phenotypically predicts VHL type 2B. We had seven patients with renal cysts. In a total follow up of 33 person years, none of these cysts progressed to renal cell carcinoma. CONCLUSION: von Hippel-Lindau disease anchored in germline mutations of the VHL gene is rare in the Norwegian population as opposed to clinical VHL disease, which appears to be relatively common in patients with apparently sporadic hemangioblastomas. There exists insufficient data regarding the natural history of patients with renal cysts, which makes it difficult to include or disregard these lesions as an entity of VHL disease. PMID- 19814754 TI - A patients state of health (disease) is a quality which defines relevance of a molecular variation, not vice versa. PMID- 19814755 TI - Cerebrospinal fluid outflow and intracranial pressure in hydrocephalic patients with external ventricular drainage. AB - BACKGROUND AND PURPOSE: The aim of this study was to monitor the 24 h cerebrospinal fluid (CSF) outflow and intracranial pressure (ICP) in hydrocephalic adult patients with external ventricular drainage (EVD). PATIENTS AND METHODS: Twelve patients (5M/7F) aged 30-69 years suffering from acute hydrocephalus requiring EVD were admitted in the neuro-intensive care unit. The CSF collecting bag was continuously weighted using a high-precision scale, the filtered output of which was fed at 1 Hz to a computer and converted to flow (Q'ext(csf)). ICP was also recorded. RESULTS: One patient was excluded because more than 80% of the Q'ext(csf) data were rejected by the system. The mean +/- SD Q'ext(csf) and ICP were respectively 7.5 +/- 3.4 ml/h (range 1.6-12.1 ml/h) and 12.4 +/- 2.7 mmHg. Two patterns of Q'ext(csf) were identified: a continuous profile and a discontinuous one with numerous bursts frequently associated with manoeuvres such as cough or chest physiotherapy. The short-term variations of Q'ext(csf) and ICP were usually unrelated. CONCLUSION: The study stresses the important inter and intra-subject variability of Q'ext(csf) in patients with EVD. The mean Q'ext(csf) is lower than the reference production rate (21 ml/h), raising the question of persistent CSF absorption and/or depressed secretion. The independent changes of Q'ext(csf) and ICP on the short term is likely to be explained by the pressure-volume characteristics of the intracranial space. PMID- 19814756 TI - Grey plumage colouration in the duck is genetically determined by the alleles on two different, interacting loci. AB - Based on the observation of a grey phenotype in the F(1) generation from a cross between two white plumage duck varieties, the white Kaiya and the white Liancheng, we hypothesized a possible interaction between two autosomal loci that determine grey plumage. Using the parental and F(1) individuals, seven testing combinations including five different F(1) intercrosses (F(2)) and two different backcrosses (BC(1) and BC(2)) were designed to test our hypothesis. It was demonstrated by chi-squared analysis that six test matings produced offspring in the expected ratios between the grey and white, with P-values ranging from 0.50 to 0.99. Another mating, where all white offspring were expected, produced 33 white individuals. These results verified that the interaction between two loci produced the grey phenotype. The C locus, which carries the recessive allele (c), was previously thought to be the only gene responsible for white plumage in the duck. This is the first report that an allele (t), carried by the white Liancheng at a different autosomal locus, also determines white plumage in ducks. Furthermore, the dominant alleles at both loci can interact with each other to produce the grey phenotype, and a new dark phenotype, observed in some F(2) individuals, can be attributed to the dosage effect of the T allele. PMID- 19814757 TI - Urticarial rush due to finasteride. PMID- 19814759 TI - Emergency department Chlamydia screening through partnership with the public health department. AB - BACKGROUND: The emergency department (ED) serves a population that may benefit from numerous screening initiatives but screening in the ED is challenging due to crowding as well as resource and time constraints. One option may be to collect specimens in the ED and then partner with the public health department (PHD) to analyze the specimens off-site and arrange follow-up treatment. OBJECTIVES: The objective was to explore the feasibility of chlamydia screening in females using a partnership model in which the ED is responsible for urine collection and the PHD is responsible for chlamydia testing, notification, and treatment. METHODS: A collaborative partnership-based chlamydia screening project was initiated at a large (90,000 visits/yr), urban, teaching ED from April 2007 to April 2008. Study information sheets were handed out to a convenience sample of eligible female patients and visitors (15-24 yr of age). Those wishing to participate provided a urine sample and follow-up contact information. The information sheet also asked if they had either lower abdominal pain or vaginal discharge (affirmative answer for either was considered symptomatic). Specimens collected in the ED were retrieved by PHD staff for off-site testing. The PHD contacted those participants testing positive using the patient provided contact information and arranged for treatment. RESULTS: Of the 633 women offered screening, 296 (47%) agreed to testing and provided samples. Of the 296 tested, 38 (12.8%) were positive for chlamydia infection, and 25 (66%) received follow-up and treatment; 13 could not be contacted through information they provided. A higher percentage of symptomatic subjects (23 of 115, or 20%) tested positive for chlamydia than asymptomatic subjects (15 of 181, or 8.3%; p < 0.01). CONCLUSIONS: This study demonstrates the feasibility of an ED-PHD partnership for chlamydia screening in young women. This model can potentially be applied to other initiatives and may improve public health screening without creating significant additional burdens for crowded EDs. PMID- 19814758 TI - Six-month follow-up of a brief intervention on self-reported safety belt use among emergency department patients. AB - OBJECTIVES: Safety belt use (SBU) reduces motor vehicle deaths by 45%. We previously reported that a brief intervention improved self-reported SBU among emergency department (ED) patients at 3 months. We sought to determine if these effects were sustained at 6 months postenrollment. METHODS: This was a prospective, randomized controlled trial of adult patients (age > or = 21 years) at an academic medical center ED from February 2006 to May 2006. Patients were systematically sampled for self-reported SBU. Those with self-reported SBU less than "always" were asked to participate. Subjects were surveyed at baseline with a nine-item series of situational SBU questions scored on a five-point Likert scale (e.g., 5 = always, 1 = never). This nine-item average comprised the mean SBU score. Subjects were randomized to a control group (CG) and an intervention group (IG). The CG received an injury prevention brochure; the IG received a brief motivation interview by a trained interventionist and the brochure. Subjects were phoned at 3 and 6 months to determine interval change in SBU scores via a standard script. Repeated-measures analysis of covariance and t-tests were used to analyze trends in mean SBU scores between groups, as well as to test mean changes in SBU scores from the 3- to 6-month intervals. RESULTS: Of 432 eligible patients, 292 enrolled (mean age = 35 years, SD +/- 11.2 years; 61% male). At baseline, there were no significant demographic differences; the IG (n = 147) and CG (n = 145) had similar mean SBU scores (2.8 vs. 2.6, p = 0.31), and 66% (n = 96 in each) completed both 3- and 6-month follow-up. The mean SBU score at 6 months in the IG was greater than in the CG group (3.6 vs. 2.9, p < 0.001), as were the mean SBU score differences from baseline (IG = 0.84 vs. CG = 0.29, p < 0.001). These differences were sustained from the 3-month interval (IG = -0.02 vs. CG = 0.06, p > 0.05). CONCLUSIONS: The previously reported finding that ED patients who received a brief motivation interview reported higher SBU scores at 3 months compared to a CG was sustained at 6-month follow-up. Although limited by self report, a brief intervention may enhance lasting SBU behavior among high-risk ED patients. PMID- 19814760 TI - Documenting the SARS epidemic in mainland China. PMID- 19814761 TI - No evidence of over-reporting of SARS in mainland China. AB - OBJECTIVE: To find out whether there was over-reporting of SARS patients in mainland China in view of the relatively low case fatality ratio in mainland China, in comparison with other affected countries and areas. METHODS: We searched PubMed for all SARS antibody detection papers (in English or Chinese language) using the keywords 'SARS' and 'antibody'. Then the resulting articles were further read through to select the SARS detection results using ELISA methods of serum samples collected at least 1 month after disease onset. A multi level logistic regression was applied to test for possible differences in the proportions positive between locations of the study. RESULTS: A total of 48 studies were identified, including 39 from mainland China and nine from elsewhere (Hong Kong, Taiwan, Canada and Vietnam). For mainland China, there was no difference between Guangdong, Beijing and other provinces in the proportions testing positive (83.0%, 85.8% and 85.4% respectively). The grand average of 84.2% seropositive was lower than the 95.1% for the countries and areas outside of mainland China combined. However, this difference was far from significant after correcting for dependency of individual tests within the same study. CONCLUSIONS: Our study showed no evidence of over-reporting of SARS in mainland China, nor in Guangdong, where the SARS epidemic started. Even if the lower seroprevalence in mainland China, relative to other affected areas, does represent actual over-reporting, then this factor can only explain a modest 10% of the lower case fatality in mainland China. PMID- 19814762 TI - The SARS outbreak in a general hospital in Tianjin, China: clinical aspects and risk factors for disease outcome. AB - OBJECTIVES: To describe clinical characteristics of severe acute respiratory syndrome (SARS) patients in a hospital in Tianjin, China, thereby comparing probable and suspected cases; to study risk factors associated with the death of cases; to describe the implementation of preventive interventions during the hospital outbreak. METHODS: Physical and haematological information was obtained from clinical records. White blood cell counts, and percentages of neutrophilic granulocytes and neutrophilic lymphocytes were measured. The service department of the hospital provided information about daily use of protective materials. Differences in clinical symptoms between probable and suspected SARS cases were tested by Fisher's exact test. Non-linear mixed modelling was used to test for differences between the haematological patterns for probable and suspected cases. Risk factors for dying among probable SARS cases were tested by logistic regression. RESULTS: The SARS outbreak started with a patient from Beijing on 15 April 2003, and spread quickly among the healthcare workers and in-patients in the hospital. In total 90 probable and 21 suspected cases were reported, with 17 deaths among them (case fatality rate 15%). Haematological patterns were significantly different between probable and suspected cases, whereas the percentages with certain clinical symptoms showed no apparent difference. Death of probable SARS cases was only significantly associated with high age and use of a respiratory machine (mainly for the most severe cases), whereas e.g. co morbidity and steroid treatment showed no impact in multivariate analysis. Stringent control measures, including distribution of huge numbers of protective materials, started on 20 April, which soon lead to a strong decrease in the incidence of new cases. After the last SARS case left the hospital on 6 June, protective materials were dispensed at substantially lower rates, but not to zero, as was the case during the first days of the outbreak. CONCLUSION: The working definition of probable SARS used during the epidemic appeared to have been fairly accurate. Many valuable lessons were learned regarding prevention of hospital spread of infection, especially the need to have sufficient protective supplies available and to implement these rigidly and at an early stage of an (threatening) epidemic. PMID- 19814763 TI - Transmission of SARS in three Chinese hospitals. AB - OBJECTIVE: To quantify the transmissibility of severe acute respiratory syndrome (SARS) in hospitals in mainland China and to assess the effectiveness of control measures. METHODS: We report key epidemiological details of three major hospital outbreaks of SARS in mainland China, and estimate the evolution of the effective reproduction number in each of the three hospitals during the course of the outbreaks. RESULTS: The three successive hospital outbreaks infected 41, 99 and 91 people of whom 37%, 60% and 70% were hospital staff. These cases resulted in 33 deaths, five of which occurred in hospital staff. In a multivariate logistic regression, age and whether or not the case was a healthcare worker (HCW) were found to be significant predictors of mortality. The estimated effective reproduction numbers (95% CI) for the three epidemics peaked at 8 (5, 11), 9 (4, 14) and 12 (7, 17). In all three hospitals the epidemics were rapidly controlled, bringing the reproduction number below one within 25, 10 and 5 days respectively. CONCLUSIONS: This work shows that in three major hospital epidemics in Beijing and Tianjin substantially higher rates of transmission were initially observed than those seen in the community. In all three cases the hospital epidemics were rapidly brought under control, with the time to successful control becoming shorter in each successive outbreak. PMID- 19814764 TI - The impact of public health control measures during the SARS epidemic in mainland China. AB - We tracked the effective reproductive number (Rt) over time to assess the impact of important public health control measures in the five most SARS-affected geographic areas in mainland China. As soon as the Chinese authorities gained full control of all activities to combat SARS, Rt decreased dramatically and consistently below one. Many control measures that seriously affected public life were implemented afterwards, i.e., when the epidemic was already dying down. PMID- 19814765 TI - Alpha-defensins increase lung fibroblast proliferation and collagen synthesis via the beta-catenin signaling pathway. AB - Alpha-defensins are released from granules of leukocytes and are implicated in inflammatory and fibrotic lung diseases. In the present study, the effects of alpha-defensins on the proliferation and collagen synthesis of lung fibroblasts were examined. We found that alpha-defensin-1 and alpha-defensin-2 induced dose dependent increases in the incorporation of 5-bromo-2'-deoxy-uridine into newly synthesized DNA in two lines of human lung fibroblasts (HFL-1 and LL-86), suggesting that alpha-defensin-1 and alpha-defensin-2 stimulate the proliferation of lung fibroblasts. alpha-defensin-1 and alpha-defensin-2 also increased collagen-I mRNA (COL1A1) levels and protein contents of collagen-I and active/dephosphorylated beta-catenin without changes in total beta-catenin protein content in lung fibroblasts (HFL-1 and LL-86). Inhibition of the beta catenin signaling pathway using quercetin prevented increases in cell proliferation and the protein content of collagen-I and active/dephosphorylated beta-catenin in lung fibroblasts, and in COL1A1 mRNA levels and collagen release into culture medium induced by alpha-defensin-1 and alpha-defensin-2. Knocking down beta-catenin using small interfering RNA technology also prevented alpha defensin-induced increases in cell proliferation and the protein content of collagen-I and active/dephosphorylated beta-catenin in lung fibroblasts, and in COL1A1 mRNA levels. Moreover, increases in the phosphorylation of glycogen synthase kinase 3beta, accumulation/activation of beta-catenin, and collagen synthesis induced by alpha-defensin-1 and alpha-defensin-2 were prevented by p38 mitogen-activated protein kinase inhibitor SB203580 and phosphoinositide 3-kinase inhibitor LY294002. These results indicate that alpha-defensin-1 and alpha defensin-2 stimulate proliferation and collagen synthesis of lung fibroblasts. The beta-catenin signaling pathway mediates alpha-defensin-induced increases in cell proliferation and collagen synthesis of lung fibroblasts. alpha-defensin induced activation of beta-catenin in lung fibroblasts might be caused by phosphorylation/inactivation of glycogen synthase kinase 3beta as a result of the activation of the p38 mitogen-activated protein kinase and phosphoinositide 3 kinase/Akt pathways. PMID- 19814766 TI - Idiopathic granulomatous and necrotising inflammatory disorders of the canine central nervous system: a review and future perspectives. AB - Granulomatous meningoencephalomyelitis, necrotizing meningoencephalitis, and necrotizing leukoencephalitis are common inflammatory conditions of the canine central nervous system. Although each disease has unique histopathological features, these canine disorders collectively seem to be aberrant immune responses directed against the central nervous system. A review of the neurological signs and general neurodiagnostic approach to canine meningoencephalitis is followed by an overview of the specific clinical and neuropathological features of granulomatous meningoencephalomyelitis, necrotizing meningoencephalitis, and necrotizing leukoencephalitis. The aetiopathogenesis of each disorder is explored including potential genetic, immunological, and environmental factors along with the current and prospective immunomodulatory therapies for meningoencephalitis. PMID- 19814767 TI - Lack of advantage of high-fibre, moderate-carbohydrate diets in dogs with stabilised diabetes. AB - OBJECTIVES: To assess the influence of high-fibre, moderate-carbohydrate diets with two levels of dietary fat, compared with a commercial diet with moderate fibre, low-carbohydrate and higher fat, on insulin requirement, glycaemic control and lipid profile of dogs with stabilised diabetes. METHODS: Twelve dogs with spontaneous diabetes mellitus were studied. Glycaemic control was evaluated by plasma fructosamine, glycosylated haemoglobin and 48-hour serial blood glucose measurements. The insulin dosage required to maintain clinical stability was also determined. Lipid profiles comprised serial measurements of plasma cholesterol, triglyceride, free glycerol and free fatty acids. Data were analysed using analysis of variance. RESULTS: There were no significant differences in insulin requirement or glycaemic control among diets. Weight loss occurred when the dogs were fed the high-fibre, moderate-carbohydrate, moderate-fat diet (P<0.002), whereas weight was maintained with the other two diets. The high-fibre, moderate carbohydrate, moderate-fat diet resulted in lower mean plasma cholesterol compared with either of the higher-fat diets (P< or =0.003), and lower mean plasma triglyceride (P=0.060), free fatty acid (P<0.001) and free glycerol (P=0.015) than the commercial diet. CLINICAL SIGNIFICANCE: For stable diabetic dogs, high-fibre, moderate-carbohydrate diets offered no significant advantage compared with a commercial diet with moderate fibre and low carbohydrate. Diets with high fibre, moderate carbohydrate and moderate fat should not be routinely recommended for dogs with thin body condition. PMID- 19814768 TI - Cardiac conotruncal malformations in a family of beagle dogs. AB - OBJECTIVES: To study forms of congenital heart disease present in a family of beagle dogs with a strong prevalence of ventricular septal defect and to document the pathological findings associated with the ventricular septal defects and investigate the mode of transmission. METHODS: The animals were investigated by physical examination, radiography, electrocardiography and ultrasonography to diagnose the presence of congenital heart disease. Some animals were diagnosed at post-mortem examination and the dead animals underwent post-mortem examination to verify the presence of ventricular septal defect. An analysis of pedigree was undertaken and two of the affected animals were mated to investigate the mode of transmission. RESULTS: Among the 28 dogs evaluated clinically or by post-mortem examination, 14 cases of ventricular septal defect were identified. The post mortem examination showed some abnormalities of the ventricular outflow region associated with malformation of conotruncal septum development. When two affected dogs were bred, congenital heart disease was present in all of the offspring. CLINICAL SIGNIFICANCE: The congenital heart disease identified in this beagle family can be classified as conotruncal malformation, and an autosomal recessive mode of inheritance was suggested by pedigree analysis. PMID- 19814769 TI - Outcome following removal of canine spindle cell tumours in first opinion practice: 104 cases. AB - OBJECTIVES: To define the outcome of a cohort of canine patients with a histological diagnosis of spindle cell tumour of soft tissue managed solely by surgery in first opinion practice. METHODS: Clinical details of 104 spindle cell sarcomas submitted to Finn Pathologists during the year 2000 were reviewed. Questionnaires were sent to the submitting veterinarians, requesting details about the tumour, surgery performed and ultimate outcome of the patient. RESULTS: The method of surgical resection was described as marginal in 45 dogs (44.2 per cent). Excision margins of 3 cm or more were described in less than 10 per cent of cases. Tumours recurred locally in 29 dogs (27.9 per cent). Eighteen dogs (21.7 per cent) died of tumour-related causes. Most deaths were unrelated to sarcoma (50 dogs, 60.2 per cent) or unknown (15 dogs, 18 per cent). The median survival time was 1013 days. Tumour size, location or degree of surgical resection were not significantly related to survival or tumour recurrence. A palpable assessment of tumour invasion into underlying tissues was significantly associated with decreased disease-free interval (P<0.0001) and survival time (P = 0.0070). CLINICAL SIGNIFICANCE: The results of this retrospective study indicate that many spindle cell tumours managed in first opinion practice exhibit a low grade biological behaviour and may respond well to more conservative surgery than current recommendations advise. PMID- 19814770 TI - Gastrointestinal foreign bodies in dogs and cats: a retrospective study of 208 cases. AB - OBJECTIVES: To establish predilection sites of obstruction and to investigate clinical factors associated with a poor outcome. METHODS: A retrospective study of 208 consecutive cases over a 48-month period from first-opinion practice. RESULTS: Overall, 91 per cent of cases recovered with higher survival rates from discrete foreign bodies (94 per cent in dogs and 100 per cent in cats) as opposed to linear foreign bodies (80 per cent in dogs and 63 per cent in cats). English bull terriers, springer spaniels, Staffordshire bull terriers, Border collies and Jack Russell terriers were over-represented. In dogs, 63 per cent of obstructions occurred in the jejunum but foreign objects were encountered at all points along the gastrointestinal tract. A longer duration of clinical signs, the presence of a linear foreign body and multiple intestinal procedures were associated with significantly increased mortality. Neither the degree of obstruction (partial or complete) nor the location of the foreign body was shown to have a significant influence on survival. CLINICAL SIGNIFICANCE: Prompt presentation, diagnosis and surgical intervention improve the outcome of gastrointestinal obstruction by foreign bodies. At surgery, the minimal number of intestinal procedures should be performed to restore the integrity of the alimentary tract. PMID- 19814771 TI - Predicting recovery of urination control in cats after sacrocaudal injury: a prospective study. AB - OBJECTIVES: To determine whether intact tail base pain sensation predicts control of urination after sacrocaudal fracture-subluxation in cats. METHODS: Twenty-one cats affected by sacrocaudal fracture-subluxation were diagnosed routinely by radiography. The ability of each cat to detect a painful stimulus applied to the tail base was tested by application of pressure using a standard surgical instrument. Cats were then hospitalised for up to 30 days and categorised according to whether they showed control of urination. A value of P<0.05 was regarded as significant. The results were analysed to produce data regarding sensitivity, specificity and the positive and negative predictive value of the test. RESULTS: All 11 cats that had intact tail base sensation showed control of urination within three days. Four of the 10 cats without tail base pain sensation did not recover control of urination by day 30. CLINICAL SIGNIFICANCE: Intact tail base sensation predicts control of urination and is therefore a useful screening test. PMID- 19814772 TI - The ageing gut: diminished symptom response to a standardized nutrient stimulus. AB - BACKGROUND The prevalence of dyspepsia and the severity of reflux symptoms decreases with advancing age. We postulate that advancing age influences sensory function and this will be associated with a diminished symptom response to a standardized meal stimulus.Our aim was to assess the influence of age on visceral sensory function. METHODS Baseline gastrointestinal symptoms and anxiety and depression were assessed in 53 healthy volunteers using validated questionnaires. After an 8-h fast, subjects received 200 mL of a standardized enteral feeding solution every 5 min up to a cumulative volume of 800 mL. After each 200 mL drink, five key symptoms were assessed (fullness, abdominal pain, retrosternal/abdominal burning, nausea and regurgitation) using a standardized instrument on visual analogue scales (0-100). The cumulative symptom score across all symptoms was calculated. KEY RESULTS Fullness was the most prominent symptom reported (79.8 +/- 9.5) followed by nausea (14.9 +/- 4.9) and pain (9.8 +/- 4.5); these three items accounted for more than 90% of the overall symptom load. The cumulative pain and nausea scores during a standardized nutrient challenge were significantly and inversely correlated with age (r = -0.43, P = 0.002 and r = 0.28, P = 0.045). Subjects >60 years of age reported significantly lower pain and nausea scores (0.9 +/- 0.9, 4.5 +/- 3.9) than did subjects <40 years (22 +/- 11.9, P = 0.002; 29.3 +/- 12, P = 0.043). CONCLUSIONS & INFERENCES Symptom responses to a standardized nutrient challenge, in particular pain and nausea, are inversely correlated with age. PMID- 19814773 TI - Enteric circular muscle dysfunction in the cystic fibrosis mouse small intestine. AB - BACKGROUND Cystic fibrosis (CF) has multiple effects on the gastrointestinal system, including altered motility. The Cftr knockout mouse model of CF has impaired small intestinal transit but the mechanism is unknown. METHODS Behaviour of circular smooth muscle was studied in an organ bath. Expression levels of prostaglandin (PG) degradative genes were measured by quantitative RT-PCR, and PGE(2) levels were measured by enzyme immunoassay. KEY RESULTS Cystic fibrosis circular muscle activity was erratic and had variable frequency of contractions, as compared to WT. The CF tissue was non-responsive to cholinergic stimulation or direct KCl depolarization. PGE(2) and PGF(2alpha) are significantly elevated in the CF mouse small intestine, and we hypothesized these contribute to impaired smooth muscle activity. After inhibition of PG synthesis, the CF circular muscle exhibited greater cholinergic responsiveness, which was reversed by exogenous PGE(2). PGF(2alpha) enhanced activity of CF tissue only after inhibition of PG synthesis. The enteric microbiota was implicated in PGE(2)-mediated dysmotility because broad spectrum antibiotic treated WT mice, which have slowed transit, exhibit impaired circular muscle activity. This was accompanied by decreased expression of PG degradative genes and increased intestinal PGE(2) levels. Furthermore, administration of oral laxative, which eradicates bacterial overgrowth and improves transit in CF mice, increased expression of PG degradative genes, decreased PGE(2) levels, and improved CF muscle activity. CONCLUSIONS & INFERENCES These results suggest that the enteric microbiota modulates PGE(2) levels in a complex manner, which affects enteric smooth muscle activity and contributes to slower small intestinal transit in CF. PMID- 19814774 TI - Does body position modify the results of oesophageal high resolution manometry? AB - BACKGROUND Oesophageal motility classification using high resolution manometry (HRM) has been established in the supine position. Nevertheless, examination in the sitting position is more physiological. Our aim was to determine if body position modifies oesophago-gastric junction (OGJ) morphology and oesophageal motility. METHODS A total of 100 patients (47 males, mean age 51 years) were included in this study. The oesophageal HRM protocol included examination in supine and sitting positions. Recordings were reviewed by two different operators. Amplitude, duration, velocity, Distal Contractile Integral (DCI) and Pressurization Front Velocity of oesophageal waves induced by swallowing were recorded. KEY RESULTS The lower oesophageal sphincter resting pressure was not significantly changed by body position. The sitting position modified the OGJ classification in 12 patients. The inter-observer agreement to classify OGJ was moderate (kappa = 0.54 and 0.46, in the supine and sitting positions respectively) while it was good to diagnose motility disorders (kappa = 0.72 and 0.83). The percentage of normal waves was lower in the sitting position in comparison with the supine position (56%vs 67%, P < 0.01). The DCI was also lower in the sitting position (1125 mmHg.s.cm vs 1639, P < 0.01) as well as the amplitude of oesophageal waves. Finally the diagnosis was concordant in both positions in 72 patients. CONCLUSIONS & INFERENCES Body position can affect OGJ morphology and oesophageal motility assessment by HRM in some patients. Normal values in the sitting position should thus be determined. Inter-observer variation for the proposed classification of OGJ morphology must also be taken into account. PMID- 19814775 TI - Is there any association between disturbed gastrointestinal visceromotor and sensory function and impaired quality of life in functional dyspepsia? AB - BACKGROUND Functional dyspepsia (FD) is now categorized into the epigastric pain syndrome (EPS) and the postprandial distress syndrome (PDS). However, the role of disturbed gastric emptying and sensory function for the reduction of health related quality of life (HRQOL) has not yet been studied in EPS and PDS. METHODS A total of 300 refractory FD patients and 450 healthy blood donors (BD) were studied. BD were stratified in subjects with (BD+) and without (BD-) concomitant FD symptoms. Gastric motor and sensory function, generic and disease-specific HRQOL [physical (PCS) and mental component summary (MCS)] and affective disorders were assessed. Twenty randomly selected BD-, 50 BD+ (36 PDS, 72%), and 110 FD (95 PDS, 86.4%) patients had additional function testing. KEY RESULTS Health-related quality of life was significantly reduced in FD patients (PCS = 40.7 +/- 8.8, MCS = 39.7 +/- 11.3, both P < 0.0001) compared to BD+ (PCS = 52.0 +/- 7.6, MCS = 49.0 +/- 9.4) and BD- (PCS = 56.0 +/- 4.3, MCS = 52.8 +/- 7.2). GET (t((1/2)), min) was significantly (both P < 0.0001) longer in FD patients (143.0 +/- 7.3) compared to BD+ (101.1 +/- 6.3) and BD- (73.8 +/- 7.6). FD patients scored significantly higher for 'pain' (P < 0.0001) and 'nausea' (P = 0.023), there was no difference for 'fullness' compared to BD. Impairment of GET was not associated with HRQOL. In FD patients, an augmented symptom response to the test meal (fullness, nausea) was associated with MCS, there was no difference between FD patients with EPS or PDS. CONCLUSIONS & INFERENCES In EPS and PDS, delayed gastric empting and altered sensory function are disease markers but not directly linked to the severity of HRQOL impairment or clinical presentation of FD. PMID- 19814776 TI - Quantitative resistance increases the durability of qualitative resistance to Leptosphaeria maculans in Brassica napus. AB - It has frequently been hypothesized that quantitative resistance increases the durability of qualitative (R-gene mediated) resistance but supporting experimental evidence is rare. To test this hypothesis, near-isogenic lines with/without the R-gene Rlm6 introduced into two Brassica napus cultivars differing in quantitative resistance to Leptosphaeria maculans were used in a 5 yr field experiment. Recurrent selection of natural fungal populations was done annually on each of the four plant genotypes, using crop residues from each genotype to inoculate separately the four series of field trials for five consecutive cropping seasons. Severity of phoma stem canker was measured on each genotype and frequencies of avirulence alleles in L. maculans populations were estimated. Recurrent selection of virulent isolates by Rlm6 in a susceptible background rendered the resistance ineffective by the third cropping season. By contrast, the resistance was still effective after 5 yr of selection by the genotype combining this gene with quantitative resistance. No significant variation in the performance of quantitative resistance alone was noted over the course of the experiment. We conclude that quantitative resistance can increase the durability of Rlm6. We recommend combining quantitative resistance with R gene mediated resistance to enhance disease control and crop production. PMID- 19814777 TI - Specificity and levels of nonhost resistance to nonadapted Blumeria graminis forms in barley. AB - The genetic basis of nonhost resistance of barley to nonadapted formae speciales of Blumeria graminis is not known, as there is no barley line that is susceptible to these nonadapted formae speciales, such as the wheat powdery mildew pathogen, Blumeria graminis f.sp. tritici (Bgt). Barley accessions with rudimentary susceptibility to an isolate of the nonadapted Bgt were identified. Those accessions were intercrossed in two cycles and two lines, called SusBgt(SC) and SusBgt(DC), with substantial susceptibility to Bgt at the seedling stage were selected. The quantitative variation among barley accessions and in the progenies after convergent crossing suggests a polygenic basis for this nonhost resistance. Both lines allowed an unusually high level of haustorium formation and colony development by Bgt. The SusBgt lines and their ancestor lines also allowed haustorium formation and conidiation by four out of seven isolates of other nonadapted B. graminis forms. Analysis of the infection process suggested that nonhost resistance factors are specific to the form and developmental stage of B. graminis. Resistances to establishment (first haustorium), colonization (subsequent haustoria) and conidiation are not associated. The lines developed will be of use in elucidating the genetic basis of nonhost resistance to Bgt in barley, and in gene expression and complementation studies on nonhost resistance. PMID- 19814778 TI - When divergent life histories hybridize: insights into adaptive life-history traits in an annual weed. AB - *Colonizing weed populations face novel selective environments, which may drive rapid shifts in life history. These shifts may be amplified when colonists are hybrids of species with divergent life histories. Selection on such phenotypically diverse hybrids may create highly fecund weeds. We measured the phenotypic variation, strength of natural selection and evolutionary response of hybrid and nonhybrid weeds. *We created F(1) hybrids of wild radish, an early flowering, small-stemmed weed, and its late-flowering, large-stemmed, crop relative (Raphanus spp.). Replicate wild and hybrid populations were established in an agricultural landscape in Michigan, USA. The consequences of three generations of natural selection were measured in a common garden experiment. *Hybrid populations experienced strong selection for larger, earlier flowering plants whereas selection was relatively weak on wild populations. Large plant size evolved two to three times faster in the hybrid populations than in wild populations, yet hybrid populations did not evolve earlier flowering. Strong selection on size and phenotypic correlations between age at reproduction and size may have limited the response of flowering phenology. *Our findings demonstrate hybridization between species with divergent life histories may catalyse the rapid evolution of certain adaptive, weedy traits while tradeoffs limit the evolution of others. PMID- 19814779 TI - Deterministic Effects Propagation Networks for reconstructing protein signaling networks from multiple interventions. AB - BACKGROUND: Modern gene perturbation techniques, like RNA interference (RNAi), enable us to study effects of targeted interventions in cells efficiently. In combination with mRNA or protein expression data this allows to gain insights into the behavior of complex biological systems. RESULTS: In this paper, we propose Deterministic Effects Propagation Networks (DEPNs) as a special Bayesian Network approach to reverse engineer signaling networks from a combination of protein expression and perturbation data. DEPNs allow to reconstruct protein networks based on combinatorial intervention effects, which are monitored via changes of the protein expression or activation over one or a few time points. Our implementation of DEPNs allows for latent network nodes (i.e. proteins without measurements) and has a built in mechanism to impute missing data. The robustness of our approach was tested on simulated data. We applied DEPNs to reconstruct the ERBB signaling network in de novo trastuzumab resistant human breast cancer cells, where protein expression was monitored on Reverse Phase Protein Arrays (RPPAs) after knockdown of network proteins using RNAi. CONCLUSION: DEPNs offer a robust, efficient and simple approach to infer protein signaling networks from multiple interventions. The method as well as the data have been made part of the latest version of the R package "nem" available as a supplement to this paper and via the Bioconductor repository. PMID- 19814780 TI - Papular xanthomas and erosive arthritis in a 3 year old girl, is this a new MRH variant? AB - Xanthomatous skin lesions and arthritis in children are not a common association. We present the case of a 3 year old girl who presented with xanthomatous lesions in the periungual region of both hands, around the nares and on her forehead, associated with significant arthritis that was clinically compatible with multicentric reticulohistiocytosis. However, pathology of the xanthomatous lesions was more suggestive of papular xanthoma, a disease that is not associated with arthritis. Based on her presentation and the negative lipid workup, she was treated for presumed multicentric reticulohistiocytosis. Multiple treatment strategies were utilized, with improvement on a combination of infliximab, methotrexate, and prednisone. We review the different diagnoses that should be considered in children with xanthomas and arthritis as well as the different pharmacologic therapies used in children with multicentric reticulohistiocytosis. PMID- 19814782 TI - Urinary estrogen metabolites and prostate cancer: a case-control study and meta analysis. AB - OBJECTIVE: To investigate prostate cancer (Pca) risk in relation to estrogen metabolism, expressed as urinary 2-hydroxyestrone (2-OHE1), 16alpha hydroxyestrone (16alpha-OHE1) and 2-OHE1 to 16alpha-OHE1 ratio. METHODS: We conducted a case-control study within the Western New York Health Cohort Study (WNYHCS) from 1996 to 2001. From January 2003 through September 2004, we completed the re-call and follow-up of 1092 cohort participants. Cases (n = 26) and controls (n = 110) were matched on age, race and recruitment period according to a 1:4 ratio. We used the unconditional logistic regression to compute crude and adjusted odds ratios (OR) and 95% confident interval (CI) of Pca in relation to 2-OHE1, 16alphaOHE1 and 2-OHE1 to 16alpha-OHE1 by tertiles of urine concentrations (stored in a biorepository for an average of 4 years). We identified age, race, education and body mass index as covariates. We also conducted a systematic review of the literature which revealed no additional studies, but we pooled the results from this study with those from a previously conducted case-control study using the DerSimonian-Laird random effects method. RESULTS: We observed a non-significant risk reduction in the highest tertile of 2 OHE1 (OR 0.72, 95% CI 0.25-2.10). Conversely, the odds in the highest tertile of 16alpha-OHE1 showed a non-significant risk increase (OR 1.76 95% CI 0.62-4.98). There was a suggestion of reduced Pca risk for men in the highest tertile of 2 OHE1 to 16alpha-OHE1 ratio (OR 0.56, 95% CI 0.19-1.68). The pooled estimates confirmed the association between an increased Pca risk and higher urinary levels of 16alpha-OHE1 (third vs. first tertile: OR 1.82, 95% CI 1.09-3.05) and the protective effect of a higher 2-OHE 1 to 16alpha-OHE1 ratio (third vs. first tertile: OR 0.53, 95% CI 0.31-0.90). CONCLUSION: Our study and the pooled results provide evidence for a differential role of the estrogen hydroxylation pathway in Pca development and encourage further study. PMID- 19814781 TI - Retinoic acid enhances skeletal muscle progenitor formation and bypasses inhibition by bone morphogenetic protein 4 but not dominant negative beta catenin. AB - BACKGROUND: Understanding stem cell differentiation is essential for the future design of cell therapies. While retinoic acid (RA) is the most potent small molecule enhancer of skeletal myogenesis in stem cells, the stage and mechanism of its function has not yet been elucidated. Further, the intersection of RA with other signalling pathways that stimulate or inhibit myogenesis (such as Wnt and BMP4, respectively) is unknown. Thus, the purpose of this study is to examine the molecular mechanisms by which RA enhances skeletal myogenesis and interacts with Wnt and BMP4 signalling during P19 or mouse embryonic stem (ES) cell differentiation. RESULTS: Treatment of P19 or mouse ES cells with low levels of RA led to an enhancement of skeletal myogenesis by upregulating the expression of the mesodermal marker, Wnt3a, the skeletal muscle progenitor factors Pax3 and Meox1, and the myogenic regulatory factors (MRFs) MyoD and myogenin. By chromatin immunoprecipitation, RA receptors (RARs) bound directly to regulatory regions in the Wnt3a, Pax3, and Meox1 genes and RA activated a beta-catenin-responsive promoter in aggregated P19 cells. In the presence of a dominant negative beta catenin/engrailed repressor fusion protein, RA could not bypass the inhibition of skeletal myogenesis nor upregulate Meox1 or MyoD. Thus, RA functions both upstream and downstream of Wnt signalling. In contrast, it functions downstream of BMP4, as it abrogates BMP4 inhibition of myogenesis and Meox1, Pax3, and MyoD expression. Furthermore, RA downregulated BMP4 expression and upregulated the BMP4 inhibitor, Tob1. Finally, RA inhibited cardiomyogenesis but not in the presence of BMP4. CONCLUSION: RA can enhance skeletal myogenesis in stem cells at the muscle specification/progenitor stage by activating RARs bound directly to mesoderm and skeletal muscle progenitor genes, activating beta-catenin function and inhibiting bone morphogenetic protein (BMP) signalling. Thus, a signalling pathway can function at multiple levels to positively regulate a developmental program and can function by abrogating inhibitory pathways. Finally, since RA enhances skeletal muscle progenitor formation, it will be a valuable tool for designing future stem cell therapies. PMID- 19814783 TI - In vivo99mTc-HYNIC-annexin V imaging of early tumor apoptosis in mice after single dose irradiation. AB - BACKGROUND: Apoptosis is a major mode of hematological tumor death after radiation. Early detection of apoptosis may be beneficial for cancer adaptive treatment. 99mTc-HYNIC-annexinV has been reported as a promising agent for in vivo apoptosis imaging. The purpose of this study is to evaluate the feasibility of in vivo99mTc-HYNIC-annexinV imaging of radiation- induced apoptosis, and to investigate its correlation with radiosensitivity. METHODS: Ten days after inoculation of tumor cells in the right upper limbs, the mice were randomly divided into two groups. The imaging group (4 mice each level, 4 dose levels) was injected with 4-8 MBq 99mTc-HYNIC-annexinV 24 hours after irradiation and imaged 1 hr post-injection, and the mice were sacrificed immediately after imaging for biodistribution analysis of annexin V. The observation group (4 mice each level, 2 dose levels) was only observed for tumor regression post-radiation. The number of apoptotic cells in a tumor was estimated with TUNEL assay. RESULTS: The 99mTc HYNIC-annexin V uptake in E14 lymphoma significantly increased as the radiation dose escalated from 0 to 8 Gy, and significantly correlated with the number of TUNEL-positive cells (r = 0.892, P < 0.001). The Annexin-V uptake and the number of TUNEL-positive cells in El4 lymphoma were significantly greater than those in S180 sarcoma. With 8 Gy, S180 sarcoma tumor showed scanty apoptosis and less shrinkage while El4 lymphoma showed remarkable apoptosis and complete remission. CONCLUSION: 99mTc-HYNIC-annexinV in vivo imaging is a feasible method to detect early radiation-induced apoptosis in different tumors, and might be predictive for radiation sensitivity. PMID- 19814784 TI - Genomic DNA k-mer spectra: models and modalities. AB - BACKGROUND: The empirical frequencies of DNA k-mers in whole genome sequences provide an interesting perspective on genomic complexity, and the availability of large segments of genomic sequence from many organisms means that analysis of k mers with non-trivial lengths is now possible. RESULTS: We have studied the k-mer spectra of more than 100 species from Archea, Bacteria, and Eukaryota, particularly looking at the modalities of the distributions. As expected, most species have a unimodal k-mer spectrum. However, a few species, including all mammals, have multimodal spectra. These species coincide with the tetrapods. Genomic sequences are clearly very complex, and cannot be fully explained by any simple probabilistic model. Yet we sought such an explanation for the observed modalities, and discovered that low-order Markov models capture this property (and some others) fairly well. CONCLUSIONS: Multimodal spectra are characterized by specific ranges of values of C+G content and of CpG dinucleotide suppression, a range that encompasses all tetrapods analyzed. Other genomes, like that of the protozoa Entamoeba histolytica, which also exhibits CpG suppression, do not have multimodal k-mer spectra. Groupings of functional elements of the human genome also have a clear modality, and exhibit either a unimodal or multimodal behaviour, depending on the two above mentioned values. PMID- 19814785 TI - Optimized sample preparation for two-dimensional gel electrophoresis of soluble proteins from chicken bursa of Fabricius. AB - BACKGROUND: Two-dimensional gel electrophoresis (2-DE) is a powerful method to study protein expression and function in living organisms and diseases. This technique, however, has not been applied to avian bursa of Fabricius (BF), a central immune organ. Here, optimized 2-DE sample preparation methodologies were constructed for the chicken BF tissue. Using the optimized protocol, we performed further 2-DE analysis on a soluble protein extract from the BF of chickens infected with virulent avibirnavirus. To demonstrate the quality of the extracted proteins, several differentially expressed protein spots selected were cut from 2 DE gels and identified by matrix-assisted laser desorption ionization time-of flight mass spectrometry (MALDI-TOF MS). RESULTS: An extraction buffer containing 7 M urea, 2 M thiourea, 2% (w/v) 3-[(3-cholamidopropyl)-dimethylammonio]-1 propanesulfonate (CHAPS), 50 mM dithiothreitol (DTT), 0.2% Bio-Lyte 3/10, 1 mM phenylmethylsulfonyl fluoride (PMSF), 20 U/ml Deoxyribonuclease I (DNase I), and 0.25 mg/ml Ribonuclease A (RNase A), combined with sonication and vortex, yielded the best 2-DE data. Relative to non-frozen immobilized pH gradient (IPG) strips, frozen IPG strips did not result in significant changes in the 2-DE patterns after isoelectric focusing (IEF). When the optimized protocol was used to analyze the spleen and thymus, as well as avibirnavirus-infected bursa, high quality 2-DE protein expression profiles were obtained. 2-DE maps of BF of chickens infected with virulent avibirnavirus were visibly different and many differentially expressed proteins were found. CONCLUSION: These results showed that method C, in concert extraction buffer IV, was the most favorable for preparing samples for IEF and subsequent protein separation and yielded the best quality 2-DE patterns. The optimized protocol is a useful sample preparation method for comparative proteomics analysis of chicken BF tissues. PMID- 19814786 TI - Ethnic background and differences in health care use: a national cross-sectional study of native Dutch and immigrant elderly in the Netherlands. AB - BACKGROUND: Immigrant elderly are a rapidly growing group in Dutch society; little is known about their health care use. This study assesses whether ethnic disparities in health care use exist and how they can be explained. Applying an established health care access model as explanatory factors, we tested health and socio-economic status, and in view of our research population we added an acculturation variable, elaborated into several sub-domains. METHODS: Cross sectional study using data from the "Social Position, Health and Well-being of Elderly Immigrants" survey, conducted in 2003 in the Netherlands. The study population consisted of first generation immigrants aged 55 years and older from the four major immigrant populations in the Netherlands and a native Dutch reference group. The average response rate to the survey was 46% (1503/3284; country of origin: Turkey n = 307, Morocco n = 284, Surinam n = 308, the Netherlands Antilles n = 300, the Netherlands n = 304). RESULTS: High ethnic disparities exist in health and health care utilisation. Immigrant elderly show a higher use of GP services and lower use of physical therapy and home care. Both self-reported health status (need factor) and language competence (part of acculturation) have high explanatory power for all types of health services utilisation; the additional impact of socio-economic status and education is low. CONCLUSION: For all health services, health disparities among all four major immigrant groups in the Netherlands translate into utilisation disparities, aggravated by lack of language competence. The resulting pattern of systematic lower health services utilisation of elderly immigrants is a challenge for health care providers and policy makers. PMID- 19814787 TI - New estimates of the number of children living with substance misusing parents: results from UK national household surveys. AB - BACKGROUND: The existing estimates of there being 250,000 - 350,000 children of problem drug users in the UK (ACMD, 2003) and 780,000 - 1.3 million children of adults with an alcohol problem (AHRSE, 2004) are extrapolations of treatment data alone or estimates from other countries, hence updated, local and broader estimates are needed. METHODS: The current work identifies profiles where the risk of harm to children could be increased by patterns of parental substance use and generates new estimates following secondary analysis of five UK national household surveys. RESULTS: The Health Survey for England (HSfE) and General Household Survey (GHS) (both 2004) generated consistent estimates - around 30% of children under-16 years (3.3 - 3.5 million) in the UK lived with at least one binge drinking parent, 8% with at least two binge drinkers and 4% with a lone (binge drinking) parent. The National Psychiatric Morbidity Survey (NPMS) indicated that in 2000, 22% (2.6 million) lived with a hazardous drinker and 6% (705,000) with a dependent drinker. The British Crime Survey (2004) and NPMS (2000) indicated that 8% (up to 978,000) of children lived with an adult who had used illicit drugs within that year, 2% (up to 256,000) with a class A drug user and 7% (up to 873,000) with a class C drug user. Around 335,000 children lived with a drug dependent user, 72,000 with an injecting drug user, 72,000 with a drug user in treatment and 108,000 with an adult who had overdosed. Elevated or cumulative risk of harm may have existed for the 3.6% (around 430,000) children in the UK who lived with a problem drinker who also used drugs and 4% (half a million) where problem drinking co-existed with mental health problems. Stronger indicators of harm emerged from the Scottish Crime Survey (2000), according to which 1% of children (around 12,000 children) had witnessed force being used against an adult in the household by their partner whilst drinking alcohol and 0.6% (almost 6000 children) whilst using drugs. CONCLUSION: Whilst harm from parental substance use is not inevitable, the number of children living with substance misusing parents exceeds earlier estimates. Widespread patterns of binge drinking and recreational drug use may expose children to sub-optimal care and substance-using role models. Implications for policy, practice and research are discussed. PMID- 19814788 TI - Nutritional and socio-economic factors associated with Plasmodium falciparum infection in children from Equatorial Guinea: results from a nationally representative survey. AB - BACKGROUND: Malaria has traditionally been a major endemic disease in Equatorial Guinea. Although parasitaemia prevalence on the insular region has been substantially reduced by vector control in the past few years, the prevalence in the mainland remains over 50% in children younger than five years. The aim of this study is to investigate the risk factors for parasitaemia and treatment seeking behaviour for febrile illness at country level, in order to provide evidence that will reinforce the EG National Malaria Control Programme. METHODS: The study was a cross-sectional survey of children 0 to 5 years old, using a multistaged, stratified, cluster-selected sample at the national level. It included a socio-demographic, health and dietary questionnaires, anthropometric measurements, and thick and thin blood smears to determine the Plasmodium infection. A multivariate logistic regression model was used to determine risk factors for parasitaemia, taking into account the cluster design. RESULTS: The overall prevalence of parasitemia was 50.9%; it was higher in rural (58.8%) compared to urban areas (44.0%, p = 0.06). Age was positively associated with parasitemia (p < 0.0001). In rural areas, risk factors included longer distance to health facilities (p = 0.01) and a low proportion of households with access to protected water in the community (p = 0.02). Having had an episode of cough in the 15 days prior to the survey was inversely related to parasitemia (p = 0.04). In urban areas, the risk factors were stunting (p = 0.005), not having taken colostrum (p = 0.01), and that someone in the household slept under a bed net (p = 0.002); maternal antimalarial medication intake during pregnancy (p = 0.003) and the household socio-economic status (p = 0.0002) were negatively associated with parasitemia. Only 55% of children with fever were taken outside their homes for care, and treatment seeking behaviour differed substantially between rural and urban populations. CONCLUSION: Results suggest that a national programme to fight malaria in Equatorial Guinea should take into account the differences between rural and urban communities in relation to risk factors for parasitaemia and treatment seeking behaviour, integrate nutrition programmes, incorporate campaigns on the importance of early treatment, and target appropriately for bed nets to reach the under-fives. PMID- 19814789 TI - Seroprevalence of HBV and HCV in primary hepatocellular carcinoma patients in Zimbabwe. AB - BACKGROUND: Primary hepatocellular carcinoma (PHC) is one of the most common cancers in Zimbabwe. Hepatitis B virus (HBV) and hepatitis C virus (HCV) are suspected to play a major role in causing this cancer. The objective of this study was to determine the seroprevalence of HBV and HCV in PHC at Parirenyatwa Referral Hospital in Zimbabwe. We evaluated the serological markers of the two viruses in patients with PHC using commercially available enzyme-linked immunosorbent kits. RESULTS: Out of the 60 patients with PHC, 48.3% were seropositive for HBV and 20.0% were seropositive for HCV. Co-infection by HCV and HBV was found in 8% of the patients. Only 13.3% of the health controls (blood donors) were positive for HBV. All the controls were negative for HCV. CONCLUSION: The high seropositivity of HBV and HCV in PHC in Zimbabwe suggested that the two viruses were a major cause of the cancer. PMID- 19814790 TI - Antimicrobial resistances do not affect colonization parameters of intestinal E. coli in a small piglet group. AB - BACKGROUND: Although antimicrobial resistance and persistence of resistant bacteria in humans and animals are major health concerns worldwide, the impact of antimicrobial resistance on bacterial intestinal colonization in healthy domestic animals has only been rarely studied. We carried out a retrospective analysis of the antimicrobial susceptibility status and the presence of resistance genes in intestinal commensal E. coli clones from clinically healthy pigs from one production unit with particular focus on effects of pheno- and/or genotypic resistance on different nominal and numerical intestinal colonization parameters. In addition, we compared the occurrence of antimicrobial resistance phenotypes and genotypes with the occurrence of virulence associated genes typical for extraintestinal pathogenic E. coli. RESULTS: In general, up to 72.1% of all E. coli clones were resistant to ampicillin, chloramphenicol, kanamycin, streptomycin, sulfamethoxazole or tetracycline with a variety of different resistance genes involved. There was no significant correlation between one of the nominal or numerical colonization parameters and the absence or presence of antimicrobial resistance properties or resistance genes. However, there were several statistically significant associations between the occurrence of single resistance genes and single virulence associated genes. CONCLUSION: The demonstrated resistance to the tested antibiotics might not play a dominant role for an intestinal colonization success in pigs in the absence of antimicrobial drugs, or cross-selection of other colonization factors e.g. virulence associated genes might compensate "the cost of antibiotic resistance". Nevertheless, resistant strains are not outcompeted by susceptible bacteria in the porcine intestine. PMID- 19814791 TI - Health service use in indigenous Sami and non-indigenous youth in North Norway: a population based survey. AB - BACKGROUND: This is the first population based study exploring health service use and ethno-cultural factors in indigenous Sami and non-Sami youth in North Norway. The first aim of the present study was to compare the frequency of health service use between Sami adolescents and their non-indigenous peers. The second aim was to explore the relationships between health service use and ethno-cultural factors, such as ethnic context, Sami self-identification, perceived discrimination and Sami language competence. Finally, we wanted to explore the relationship between use of health services and emotional and behavioural problems. METHOD: The Norwegian Arctic Adolescent Health Study was conducted among 10th graders (15-16 years old) in junior high schools in North Norway. The sample consisted of 4,449 adolescents, of whom 450 (10.1%) were indigenous Sami and 3,999 (89.9%) were non-Sami. RESULTS: Sami and non-Sami youth used all health services with equal frequency. However, several ethno-cultural factors were found to influence health service use. Sami youth in more assimilated ethnic contexts used general practitioners more than non-Sami youth. Youth with Sami self identification had a higher probability of using the school health service compared with other youth. Ethnic barriers to health service use were also identified. Sami speaking youth with a high degree of perceived discrimination had lower probability of using school health services than non-Sami speaking youth. Sami youth with conduct problems were less likely than non-Sami to use psychologist/psychiatrist. The present study demonstrated a relationship between health need and actual health service use. CONCLUSION: Culture-specific factors influenced the help-seeking process in indigenous youth; some factors acted as barriers against health service use and other factors increased the probability of health service use. PMID- 19814792 TI - Characterization of unknown genetic modifications using high throughput sequencing and computational subtraction. AB - BACKGROUND: When generating a genetically modified organism (GMO), the primary goal is to give a target organism one or several novel traits by using biotechnology techniques. A GMO will differ from its parental strain in that its pool of transcripts will be altered. Currently, there are no methods that are reliably able to determine if an organism has been genetically altered if the nature of the modification is unknown. RESULTS: We show that the concept of computational subtraction can be used to identify transgenic cDNA sequences from genetically modified plants. Our datasets include 454-type sequences from a transgenic line of Arabidopsis thaliana and published EST datasets from commercially relevant species (rice and papaya). CONCLUSION: We believe that computational subtraction represents a powerful new strategy for determining if an organism has been genetically modified as well as to define the nature of the modification. Fewer assumptions have to be made compared to methods currently in use and this is an advantage particularly when working with unknown GMOs. PMID- 19814793 TI - Electrical muscle stimulation preserves the muscle mass of critically ill patients: a randomized study. AB - INTRODUCTION: Critically ill patients are characterized by increased loss of muscle mass, partially attributed to sepsis and multiple organ failure, as well as immobilization. Recent studies have shown that electrical muscle stimulation (EMS) may be an alternative to active exercise in chronic obstructive pulmonary disease (COPD) and chronic heart failure (CHF) patients with myopathy. The aim of our study was to investigate the EMS effects on muscle mass preservation of critically ill patients with the use of ultrasonography (US). METHODS: Forty-nine critically ill patients (age: 59 +/- 21 years) with an APACHE II admission score >or=13 were randomly assigned after stratification upon admission to receive daily EMS sessions of both lower extremities (EMS-group) or to the control group (control group). Muscle mass was evaluated with US, by measuring the cross sectional diameter (CSD) of the vastus intermedius and the rectus femoris of the quadriceps muscle. RESULTS: Twenty-six patients were finally evaluated. Right rectus femoris and right vastus intermedius CSD decreased in both groups (EMS group: from 1.42 +/- 0.48 to 1.31 +/- 0.45 cm, P = 0.001 control group: from 1.59 +/- 0.53 to 1.37 +/- 0.5 cm, P = 0.002; EMS group: from 0.91 +/- 0.39 to 0.81 +/- 0.38 cm, P = 0.001 control group: from 1.40 +/- 0.64 to 1.11 +/- 0.56 cm, P = 0.004, respectively). However, the CSD of the right rectus femoris decreased significantly less in the EMS group (-0.11 +/- 0.06 cm, -8 +/- 3.9%) as compared to the control group (-0.21 +/- 0.10 cm, -13.9 +/- 6.4%; P < 0.05) and the CSD of the right vastus intermedius decreased significantly less in the EMS group (-0.10 +/- 0.05 cm, -12.5 +/- 7.4%) as compared to the control group (-0.29 +/- 0.28 cm, -21.5 +/- 15.3%; P < 0.05). CONCLUSIONS: EMS is well tolerated and seems to preserve the muscle mass of critically ill patients. The potential use of EMS as a preventive and rehabilitation tool in ICU patients with polyneuromyopathy needs to be further investigated. TRIAL REGISTRATION: clinicaltrials.gov: NCT00882830. PMID- 19814794 TI - A compiled and systematic reference map of nucleosome positions across the Saccharomyces cerevisiae genome. AB - Nucleosomes have position-specific functions in controlling gene expression. A complete systematic genome-wide reference map of absolute and relative nucleosome positions is needed to minimize potential confusion when referring to the function of individual nucleosomes (or nucleosome-free regions) across datasets. We compiled six high-resolution genome-wide maps of Saccharomyces cerevisiae nucleosome positions from multiple labs and detection platforms, and report new insights. Data downloads, reference position assignment software, queries, and a visualization browser are available online http://atlas.bx.psu.edu/. PMID- 19814795 TI - Oxidative stress and S-100B protein in children with bacterial meningitis. AB - BACKGROUND: Bacterial meningitis is often associated with cerebral compromise which may be responsible for neurological sequelae in nearly half of the survivors. Little is known about the mechanisms of CNS involvement in bacterial meningitis. Several studies have provided substantial evidence for the key role of nitric oxide (NO) and reactive oxygen species in the complex pathophysiology of bacterial meningitis. METHODS: In the present study, serum and CSF levels of NO, lipid peroxide (LPO) (mediators for oxidative stress and lipid peroxidation); total thiol, superoxide dismutase (SOD) (antioxidant mediators) and S-100B protein (mediator of astrocytes activation and injury), were investigated in children with bacterial meningitis (n = 40). Albumin ratio (CSF/serum) is a marker of blood-CSF barriers integrity, while mediator index (mediator ratio/albumin ratio) is indicative of intrathecal synthesis. RESULTS: Compared to normal children (n = 20), patients had lower serum albumin but higher NO, LPO, total thiol, SOD and S-100B. The ratios and indices of NO and LPO indicate blood CSF barriers dysfunction, while the ratio of S-100B indicates intrathecal synthesis. Changes were marked among patients with positive culture and those with neurological complications. Positive correlation was found between NO index with CSF WBCs (r = 0.319, p < 0.05); CSF-LPO with CSF-protein (r = 0.423, p < 0.01); total thiol with LPO indices (r = 0.725, p < 0.0001); S-100B and Pediatric Glasow Coma Scores (0.608, p < 0.0001); CSF-LPO with CSF-S-100B (r = 0.482, p < 0.002); serum-total thiol with serum S-100B (r = 0.423, p < 0.01). CONCLUSION: This study suggests that loss of integrity of brain-CSF barriers, oxidative stress and S-100B may contribute to the severity and neurological complications of bacterial meningitis. PMID- 19814796 TI - AI-2 does not function as a quorum sensing molecule in Campylobacter jejuni during exponential growth in vitro. AB - BACKGROUND: Campylobacter jejuni contains a homologue of the luxS gene shown to be responsible for the production of the signalling molecule autoinducer-2 (AI-2) in Vibrio harveyi and Vibrio cholerae. The aim of this study was to determine whether AI-2 acted as a diffusible quorum sensing signal controlling C. jejuni gene expression when it is produced at high levels during mid exponential growth phase. RESULTS: AI-2 activity was produced by the parental strain NCTC 11168 when grown in rich Mueller-Hinton broth (MHB) as expected, but interestingly was not present in defined Modified Eagles Medium (MEM-alpha). Consistent with previous studies, the luxS mutant showed comparable growth rates to the parental strain and exhibited decreased motility halos in both MEM-alpha and MHB. Microarray analysis of genes differentially expressed in wild type and luxS mutant strains showed that many effects on mRNA transcript abundance were dependent on the growth medium and linked to metabolic functions including methionine metabolism. Addition of exogenously produced AI-2 to the wild type and the luxS mutant, growing exponentially in either MHB or MEM-alpha did not induce any transcriptional changes as analysed by microarray. CONCLUSION: Taken together these results led us to conclude that there is no evidence for the role of AI-2 in cell-to-cell communication in C. jejuni strain NCTC 11168 under the growth conditions used, and that the effects of the luxS mutation on the transcriptome are related to the consequential loss of function in the activated methyl cycle. PMID- 19814797 TI - Clinical effectiveness and cost-effectiveness of pegvisomant for the treatment of acromegaly: a systematic review and economic evaluation. AB - BACKGROUND: Acromegaly, an orphan disease usually caused by a benign pituitary tumour, is characterised by hyper-secretion of growth hormone (GH) and insulin like growth factor I (IGF-1). It is associated with reduced life expectancy, cardiovascular problems, a variety of insidiously progressing detrimental symptoms and metabolic malfunction. Treatments include surgery, radiotherapy and pharmacotherapy. Pegvisomant (PEG) is a genetically engineered GH analogue licensed as a third or fourth line option when other treatments have failed to normalise IGF-1 levels. METHODS: Evidence about effectiveness and cost effectiveness of PEG was systematically reviewed. Data were extracted from published studies and used for a narrative synthesis of evidence. A decision analytical economic model was identified and modified to assess the cost effectiveness of PEG. RESULTS: One RCT and 17 non-randomised studies were reviewed for effectiveness. PEG substantially reduced and rapidly normalised IGF 1 levels in the majority of patients, approximately doubled GH levels, and improved some of the signs and symptoms of the disease. Tumour size was unaffected at least in the short term. PEG had a generally safe adverse event profile but a few patients were withdrawn from treatment because of raised liver enzymes. An economic model was identified and adapted to estimate the lower limit for the cost-effectiveness of PEG treatment versus standard care. Over a 20 year time horizon the incremental cost-effectiveness ratio was pound81,000/QALY and pound212,000/LYG. To reduce this to pound30K/QALY would require a reduction in drug cost by about one third. CONCLUSION: PEG is highly effective for improving patients' IGF-1 level. Signs and symptoms of disease improve but evidence is lacking about long term effects on improved signs and symptoms of disease, quality of life, patient compliance and safety. Economic evaluation indicated that if current standards (UK) for determining cost-effectiveness of therapies were to be applied to PEG it would be considered not to represent good value for money. PMID- 19814798 TI - Eps15: a multifunctional adaptor protein regulating intracellular trafficking. AB - Over expression of receptor tyrosine kinases is responsible for the development of a wide variety of malignancies. Termination of growth factor signaling is primarily determined by the down regulation of active growth factor/receptor complexes. In recent years, considerable insight has been gained in the endocytosis and degradation of growth factor receptors. A crucial player in this process is the EGFR Protein tyrosine kinase Substrate #15, or Eps15. This protein functions as a scaffolding adaptor protein and is involved both in secretion and endocytosis. Eps15 has been shown to bind to AP-1 and AP-2 complexes, to bind to inositol lipids and to several other proteins involved in the regulation of intracellular trafficking. In addition, Eps15 has been detected in the nucleus of mammalian cells. Activation of growth factor receptors induces tyrosine phosphorylation and mono-ubiquitination of Eps15. The role of these post translational modifications of Eps15 is still a mystery. It is proposed that Eps15 and its family members Eps15R and Eps15b are involved in the regulation of membrane morphology, which is required for intracellular vesicle formation and trafficking. PMID- 19814799 TI - Influence of the RNase H domain of retroviral reverse transcriptases on the metal specificity and substrate selection of their polymerase domains. AB - Reverse transcriptases from HIV-1 and MuLV respectively prefer Mg2+ and Mn2+ for their polymerase activity, with variable fidelity, on both RNA and DNA templates. The function of the RNase H domain with respect to these parameters is not yet understood. To evaluate this function, two chimeric enzymes were constructed by swapping the RNase H domains between HIV-1 RT and MuLV RT. Chimeric HIV-1 RT, having the RNase H domain of MuLV RT, inherited the divalent cation preference characteristic of MuLV RT on the DNA template with no significant change on the RNA template. Chimeric MuLV RT, likewise partially inherited the metal ion preference of HIV-1 RT. Unlike the wild-type MuLV RT, chimeric MuLV RT is able to use both Mn.dNTP and Mg.dNTP on the RNA template with similar efficiency, while a 30-fold higher preference for Mn.dNTP was seen on the DNA template. The metal preferences for the RNase H activity of chimeric HIV-1 RT and chimeric MuLV RT were, respectively, Mn2+ and Mg2+, a property acquired through their swapped RNase H domains. Chimeric HIV-1 RT displayed higher fidelity and discrimination against rNTPs than against dNTPs substrates, a property inherited from MuLV RT. The overall fidelity of the chimeric MuLV RT was decreased in comparison to the parental MuLV RT, suggesting that the RNase H domain profoundly influences the function of the polymerase domain. PMID- 19814800 TI - A stochastic context free grammar based framework for analysis of protein sequences. AB - BACKGROUND: In the last decade, there have been many applications of formal language theory in bioinformatics such as RNA structure prediction and detection of patterns in DNA. However, in the field of proteomics, the size of the protein alphabet and the complexity of relationship between amino acids have mainly limited the application of formal language theory to the production of grammars whose expressive power is not higher than stochastic regular grammars. However, these grammars, like other state of the art methods, cannot cover any higher order dependencies such as nested and crossing relationships that are common in proteins. In order to overcome some of these limitations, we propose a Stochastic Context Free Grammar based framework for the analysis of protein sequences where grammars are induced using a genetic algorithm. RESULTS: This framework was implemented in a system aiming at the production of binding site descriptors. These descriptors not only allow detection of protein regions that are involved in these sites, but also provide insight in their structure. Grammars were induced using quantitative properties of amino acids to deal with the size of the protein alphabet. Moreover, we imposed some structural constraints on grammars to reduce the extent of the rule search space. Finally, grammars based on different properties were combined to convey as much information as possible. Evaluation was performed on sites of various sizes and complexity described either by PROSITE patterns, domain profiles or a set of patterns. Results show the produced binding site descriptors are human-readable and, hence, highlight biologically meaningful features. Moreover, they achieve good accuracy in both annotation and detection. In addition, findings suggest that, unlike current state-of-the-art methods, our system may be particularly suited to deal with patterns shared by non-homologous proteins. CONCLUSION: A new Stochastic Context Free Grammar based framework has been introduced allowing the production of binding site descriptors for analysis of protein sequences. Experiments have shown that not only is this new approach valid, but produces human-readable descriptors for binding sites which have been beyond the capability of current machine learning techniques. PMID- 19814801 TI - KEGGconverter: a tool for the in-silico modelling of metabolic networks of the KEGG Pathways database. AB - BACKGROUND: The KEGG Pathway database is a valuable collection of metabolic pathway maps. Nevertheless, the production of simulation capable metabolic networks from KEGG Pathway data is a challenging complicated work, regardless the already developed tools for this scope. Originally used for illustration purposes, KEGG Pathways through KGML (KEGG Markup Language) files, can provide complete reaction sets and introduce species versioning, which offers advantages for the scope of cellular metabolism simulation modelling. In this project, KEGGconverter is described, implemented also as a web-based application, which uses as source KGML files, in order to construct integrated pathway SBML models fully functional for simulation purposes. RESULTS: A case study of the integration of six human metabolic pathways from KEGG depicts the ability of KEGGconverter to automatically produce merged and converted to SBML fully functional pathway models, enhanced with default kinetics. The suitability of the developed tool is demonstrated through a comparison with other state-of-the art relevant software tools for the same data fusion and conversion tasks, thus illustrating the problems and the relevant workflows. Moreover, KEGGconverter permits the inclusion of additional reactions in the resulting model which represent flux cross-talk with neighbouring pathways, providing in this way improved simulative accuracy. These additional reactions are introduced by exploiting relevant semantic information for the elements of the KEGG Pathways database. The architecture and functionalities of the web-based application are presented. CONCLUSION: KEGGconverter is capable of producing integrated analogues of metabolic pathways appropriate for simulation tasks, by inputting only KGML files. The web application acts as a user friendly shell which transparently enables the automated biochemically correct pathway merging, conversion to SBML format, proper renaming of the species, and insertion of default kinetic properties for the pertaining reactions. The tool is available at: http://www.grissom.gr/keggconverter. PMID- 19814802 TI - Diesel exhaust particles modulate the tight junction protein occludin in lung cells in vitro. AB - BACKGROUND: Using an in vitro triple cell co-culture model consisting of human epithelial cells (16HBE14o-), monocyte-derived macrophages and dendritic cells, it was recently demonstrated that macrophages and dendritic cells create a transepithelial network between the epithelial cells to capture antigens without disrupting the epithelial tightness. The expression of the different tight junction proteins in macrophages and dendritic cells, and the formation of tight junction-like structures with epithelial cells has been demonstrated. Immunofluorescent methods combined with laser scanning microscopy and quantitative real-time polymerase chain reaction were used to investigate if exposure to diesel exhaust particles (DEP) (0.5, 5, 50, 125 mug/ml), for 24 h, can modulate the expression of the tight junction mRNA/protein of occludin, in all three cell types. RESULTS: Only the highest dose of DEP (125 mug/ml) seemed to reduce the occludin mRNA in the cells of the defence system however not in epithelial cells, although the occludin arrangement in the latter cell type was disrupted. The transepithelial electrical resistance was reduced in epithelial cell mono-cultures but not in the triple cell co-cultures, following exposure to high DEP concentration. Cytotoxicity was not found, in either epithelial mono cultures nor in triple cell co-cultures, after exposure to the different DEP concentrations. CONCLUSION: We concluded that high concentrations of DEP (125 mug/ml) can modulate the tight junction occludin mRNA in the cells of the defence system and that those cells play an important role maintaining the epithelial integrity following exposure to particulate antigens in lung cells. PMID- 19814803 TI - Identification of glucocorticoid-induced leucine zipper as a key regulator of tumor cell proliferation in epithelial ovarian cancer. AB - BACKGROUND: Little is known about the molecules that contribute to tumor progression of epithelial ovarian cancer (EOC), currently a leading cause of mortality from gynecological malignancies. Glucocorticoid-Induced Leucine Zipper (GILZ), an intracellular protein widely expressed in immune tissues, has been reported in epithelial tissues and controls some of key signaling pathways involved in tumorigenesis. However, there has been no report on GILZ in EOC up to now. The objectives of the current study were to examine the expression of GILZ in EOC and its effect on tumor cell proliferation. RESULTS: GILZ expression was measured by immunohistochemical staining in tissue sections from 3 normal ovaries, 7 benign EOC and 50 invasive EOC. GILZ was not detected on the surface epithelium of normal ovaries and benign tumors. In contrast, it was expressed in the cytoplasm of tumor cells in 80% EOC specimens. GILZ immunostaining scores correlated positively to the proliferation marker Ki-67 (Spearman test in univariate analysis, P < 0.00001, r = 0.56). They were also higher in tumor cells containing large amounts of phosphorylated protein kinase B (p-AKT) (unpaired t test, P < 0.0001). To assess the effect of GILZ on proliferation and AKT activation, we used the BG-1 cell line derived from ovarian tumor cells as a cellular model. GILZ expression was either enhanced by stable transfection or decreased by the use of small interfering (si) RNA targeting GILZ. We found that GILZ increased cell proliferation, phospho-AKT cellular content and AKT kinase activity. Further, GILZ upregulated cyclin D1 and phosphorylated retinoblastoma (p-Rb), downregulated cyclin-dependent kinase inhibitor p21, and promoted the entry into S phase of cell cycle. CONCLUSION: The present study is the first to identify GILZ as a molecule produced by ovarian cancer cells that promotes cell cycle progression and proliferation. Our findings clearly indicate that GILZ activates AKT, a crucial signaling molecule in tumorigenesis. GILZ thus appears as a potential key molecule in EOC. PMID- 19814804 TI - Variation in the human soluble epoxide hydrolase gene and risk of restenosis after percutaneous coronary intervention. AB - BACKGROUND: Restenosis represents the major limiting factor for the long-term efficacy of percutaneous coronary intervention (PCI). Several genetic factors involved in the regulation of the vascular system have been described to play a role in the pathogenesis of restenosis. We investigated whether the EPHX2 K55R polymorphism, previously linked to significantly higher risk for coronary heart disease (CHD), was associated with the occurrence of restenosis after PCI. The association with incident CHD should have been confirmed and a potential correlation of the EPHX2 K55R variant to an increased risk of hypertension was analysed. METHODS: An overall cohort of 706 patients was studied: This cohort comprised of 435 CHD patients who had undergone successful PCI. Follow-up coronary angiography in all patients was performed 6 months after intervention. Another 271 patients in whom CHD had been excluded by coronary angiography served as controls. From each patient EDTA-blood was drawn at the baseline ward round. Genomic DNA was extracted from these samples and genotyping was performed by real time PCR and subsequent melting curve analysis. RESULTS: In CHD patients 6 month follow-up coronary angiography revealed a restenosis rate of 29.4%, classified as late lumen loss as well as lumen re-narrowing >or= 50%.Statistical analysis showed an equal genotype distribution in restenosis patients and non-restenosis patients (A/A 82.0% and A/G + G/G 18.0% versus A/A 82.1% and A/G + G/G 17.9%). Moreover, neither a significant difference in the genotype distribution of CHD patients and controls nor an association with increased risk of hypertension was found. CONCLUSION: The results of the present study indicate that the EPHX2 K55R polymorphism is not associated with restenosis after PCI, with incidence of CHD, or with an increased risk of hypertension and therefore, can not serve as a predictor for risk of CHD or restenosis after PCI. PMID- 19814805 TI - Polymorphism of viral dsRNA in Xanthophyllomyces dendrorhous strains isolated from different geographic areas. AB - BACKGROUND: Strains of the astaxanthin producing yeast Xanthophyllomyces dendrorhous have been isolated from different cold regions around the earth, and the presence of double stranded RNA (dsRNA) elements was described in some isolates. This kind of viruses is widely distributed among yeasts and filamentous fungi and, although generally are cryptic in function, their studies have been a key factor in the knowledge of important fungi. In this work, the characterization and genetic relationships among dsRNA elements were determined in strains representatives of almost all regions of the earth where X. dendrorhous have been isolated. RESULTS: Almost all strains of X. dendrorhous analyzed carry one, two or four dsRNA elements, of molecular sizes in the range from 0.8 to 5.0 kb. Different dsRNA-patterns were observed in strains with different geographic origin, being L1 (5.0 kb) the common dsRNA element. By hybridization assays a high genomic polymorphism was observed among L1 dsRNAs of different X. dendrorhous strains. Contrary, hybridization was observed between L1 and L2 dsRNAs of strains from same or different regions, while the dsRNA elements of minor sizes (M, S1, and S2) present in several strains did not show hybridization with neither L1 or L2 dsRNAs. Along the growth curve of UCD 67-385 (harboring four dsRNAs) an increase of L2 relative to L1 dsRNA was observed, while the S1/L1 ratio remains constant, as well as the M/L1 ratio of Patagonian strain. Strains cured of S2 dsRNA were obtained by treatment with anisomycin, and comparison of its dsRNA contents with uncured strain, revealed an increase of L1 dsRNA while the L2 and S1 dsRNA remain unaltered. CONCLUSION: The dsRNA elements of X. dendrorhous are highly variable in size and sequence, and the dsRNA pattern is specific to the geographic region of isolation. Each L1 and L2 dsRNA are viral elements able to self replicate and to coexist into a cell, and L1 and S2 dsRNAs elements could be part of a helper/satellite virus system in X. dendrorhous. PMID- 19814806 TI - Effects of isoflavone-supplemented soy yogurt on lipid parameters and atherosclerosis development in hypercholesterolemic rabbits: a randomized double blind study. AB - BACKGROUND: There is increasing interest in natural treatments to control dyslipidemia and reduce the risk of cardiovascular disease. Previous studies have demonstrated the beneficial effects of soy yogurt fermented with Enterococcus faecium CRL 183 and of dietary isoflavones on the lipid profile. The purpose of the present study was to investigate the effects of isoflavone-supplemented soy yogurt, fermented with E. faecium CRL183, on lipid parameters and atherosclerosis development in rabbits with induced hypercholesterolemia. METHODS: Forty-eight rabbits were randomly assigned to eight groups fed on the following diets for 60 days: C - control; IY - isoflavone-supplemented soy yogurt; H - hypercholesterolemic (1.0% cholesterol wt/wt diet); HY - hypercholesterolemic plus soy yogurt; HIY - hypercholesterolemic plus isoflavone-supplemented soy yogurt; HP - hypercholesterolemic plus placebo; HI - hypercholesterolemic plus isoflavone and HE - hypercholesterolemic plus pure culture of E. faecium CRL 183. Serum lipids and autoantibodies against oxLDL (oxLDL Ab) were analyzed on days 0, 30 and 60 of the treatment and the atherosclerotic lesions were quantified at the end of the experiment. RESULTS: Soy yogurt, soy yogurt supplemented with isoflavones and placebo promoted significant reductions in total cholesterol level (38.1%, 27.0% and 26.6%, respectively). Significant increases in serum HDL C concentration relative to group H were detected in animals that ingested soy yogurt, with or without the isoflavone supplement (55.2%), E. faecium culture (43.3%) or placebo (35.8%). Intake of soy yogurt and soy yogurt supplemented with isoflavones prevented the rise of oxLDL Ab during the study period. The extent of atherosclerosis in the thoracic and abdominal aortas was reduced in the HIY, HY and HP groups. However, when the whole aorta was analyzed, animals treated with soy yogurt supplemented with isoflavones exhibited the greatest reduction (51.4%, P < 0.05) in atherosclerotic lesion area, compared to group H. CONCLUSION: Soy yogurt could be consumed as an alternative means of reducing the risk of cardiovascular disease by improving the lipid profile and inhibiting oxLDL Ab formation. Our findings also suggest that isoflavone supplementation may enhance the antiatherosclerotic effect of soy yogurt. PMID- 19814808 TI - Blood feeding by the Rocky Mountain spotted fever vector, Dermacentor andersoni, induces interleukin-4 expression by cognate antigen responding CD4+ T cells. AB - BACKGROUND: Tick modulation of host defenses facilitates both blood feeding and pathogen transmission. Several tick species deviate host T cell responses toward a Th2 cytokine profile. The majority of studies of modulation of T cell cytokine expression by ticks were performed with lymphocytes from infested mice stimulated in vitro with polyclonal T cell activators. Those reports did not examine tick modulation of antigen specific responses. We report use of a transgenic T cell receptor (TCR) adoptive transfer model reactive with influenza hemagglutinin peptide (110-120) to examine CD4+ T cell intracellular cytokine responses during infestation with the metastriate tick, Dermacentor andersoni, or exposure to salivary gland extracts. RESULTS: Infestation with pathogen-free D. andersoni nymphs or administration of an intradermal injection of female or male tick salivary gland extract induced significant increases of IL-4 transcripts in skin and draining lymph nodes of BALB/c mice as measured by quantitative real-time RT PCR. Furthermore, IL-10 transcripts were significantly increased in skin while IL 2 and IFN-gamma transcripts were not significantly changed by tick feeding or intradermal injection of salivary gland proteins, suggesting a superimposed Th2 response. Infestation induced TCR transgenic CD4+ T cells to divide more frequently as measured by CFSE dilution, but more notably these CD4+ T cells also gained the capacity to express IL-4. Intracellular levels of IL-4 were significantly increased. A second infestation administered 14 days after a primary exposure to ticks resulted in partially reduced CFSE dilution with no change in IL-4 expression when compared to one exposure to ticks. Intradermal inoculation of salivary gland extracts from both male and female ticks also induced IL-4 expression. CONCLUSION: This is the first report of the influence of a metastriate tick on the cytokine profile of antigen specific CD4+ T cells. Blood feeding by D. andersoni pathogen-free nymphs or intradermal injection of salivary gland extracts programs influenza hemagglutinin influenza peptide specific TCR transgenic CD4+ T cells to express IL-4. PMID- 19814807 TI - Dynamic regulation of the endocannabinoid system: implications for analgesia. AB - The analgesic effects of cannabinoids are well documented, but these are often limited by psychoactive side-effects. Recent studies indicate that the endocannabinoid system is dynamic and altered under different pathological conditions, including pain states. Changes in this receptor system include altered expression of receptors, differential synthetic pathways for endocannabinoids are expressed by various cell types, multiple pathways of catabolism and the generation of biologically active metabolites, which may be engaged under different conditions. This review discusses the evidence that pain states alter the endocannabinoid receptor system at key sites involved in pain processing and how these changes may inform the development of cannabinoid-based analgesics. PMID- 19814809 TI - Evaluating geographic imputation approaches for zip code level data: an application to a study of pediatric diabetes. AB - BACKGROUND: There is increasing interest in the study of place effects on health, facilitated in part by geographic information systems. Incomplete or missing address information reduces geocoding success. Several geographic imputation methods have been suggested to overcome this limitation. Accuracy evaluation of these methods can be focused at the level of individuals and at higher group levels (e.g., spatial distribution). METHODS: We evaluated the accuracy of eight geo-imputation methods for address allocation from ZIP codes to census tracts at the individual and group level. The spatial apportioning approaches underlying the imputation methods included four fixed (deterministic) and four random (stochastic) allocation methods using land area, total population, population under age 20, and race/ethnicity as weighting factors. Data included more than 2,000 geocoded cases of diabetes mellitus among youth aged 0-19 in four U.S. regions. The imputed distribution of cases across tracts was compared to the true distribution using a chi-squared statistic. RESULTS: At the individual level, population-weighted (total or under age 20) fixed allocation showed the greatest level of accuracy, with correct census tract assignments averaging 30.01% across all regions, followed by the race/ethnicity-weighted random method (23.83%). The true distribution of cases across census tracts was that 58.2% of tracts exhibited no cases, 26.2% had one case, 9.5% had two cases, and less than 3% had three or more. This distribution was best captured by random allocation methods, with no significant differences (p-value > 0.90). However, significant differences in distributions based on fixed allocation methods were found (p value < 0.0003). CONCLUSION: Fixed imputation methods seemed to yield greatest accuracy at the individual level, suggesting use for studies on area-level environmental exposures. Fixed methods result in artificial clusters in single census tracts. For studies focusing on spatial distribution of disease, random methods seemed superior, as they most closely replicated the true spatial distribution. When selecting an imputation approach, researchers should consider carefully the study aims. PMID- 19814810 TI - Modulation of pathogen-induced CCL20 secretion from HT-29 human intestinal epithelial cells by commensal bacteria. AB - BACKGROUND: Human intestinal epithelial cells (IECs) secrete the chemokine CCL20 in response to infection by various enteropathogenic bacteria or exposure to bacterial flagellin. CCL20 recruits immature dendritic cells and lymphocytes to target sites. Here we investigated IEC responses to various pathogenic and commensal bacteria as well as the modulatory effects of commensal bacteria on pathogen-induced CCL20 secretion. HT-29 human IECs were incubated with commensal bacteria (Bifidobacterium infantis or Lactobacillus salivarius), or with Salmonella typhimurium, its flagellin, Clostridium difficile, Mycobacterium paratuberculosis, or Mycobacterium smegmatis for varying times. In some studies, HT-29 cells were pre-treated with a commensal strain for 2 hr prior to infection or flagellin stimulation. CCL20 and interleukin (IL)-8 secretion and nuclear factor (NF)-kappaB activation were measured using enzyme-linked immunosorbent assays. RESULTS: Compared to untreated cells, S. typhimurium, C. difficile, M. paratuberculosis, and flagellin activated NF-kappaB and stimulated significant secretion of CCL20 and IL-8 by HT-29 cells. Conversely, B. infantis, L. salivarius or M. smegmatis did not activate NF-kappaB or augment CCL20 or IL-8 production. Treatment with B. infantis, but not L. salivarius, dose-dependently inhibited the baseline secretion of CCL20. In cells pre-treated with B. infantis, C. difficile-, S. typhimurium-, and flagellin-induced CCL20 were significantly attenuated. B. infantis did not limit M. Paratuberculosis-induced CCL20 secretion. CONCLUSION: This study is the first to demonstrate that a commensal strain can attenuate CCL20 secretion in HT-29 IECs. Collectively, the data indicate that M. paratuberculosis may mediate mucosal damage and that B. infantis can exert immunomodulatory effects on IECs that mediate host responses to flagellin and flagellated enteric pathogens. PMID- 19814811 TI - Riboswitch detection using profile hidden Markov models. AB - BACKGROUND: Riboswitches are a type of noncoding RNA that regulate gene expression by switching from one structural conformation to another on ligand binding. The various classes of riboswitches discovered so far are differentiated by the ligand, which on binding induces a conformational switch. Every class of riboswitch is characterized by an aptamer domain, which provides the site for ligand binding, and an expression platform that undergoes conformational change on ligand binding. The sequence and structure of the aptamer domain is highly conserved in riboswitches belonging to the same class. We propose a method for fast and accurate identification of riboswitches using profile Hidden Markov Models (pHMM). Our method exploits the high degree of sequence conservation that characterizes the aptamer domain. RESULTS: Our method can detect riboswitches in genomic databases rapidly and accurately. Its sensitivity is comparable to the method based on the Covariance Model (CM). For six out of ten riboswitch classes, our method detects more than 99.5% of the candidates identified by the much slower CM method while being several hundred times faster. For three riboswitch classes, our method detects 97-99% of the candidates relative to the CM method. Our method works very well for those classes of riboswitches that are characterized by distinct and conserved sequence motifs. CONCLUSION: Riboswitches play a crucial role in controlling the expression of several prokaryotic genes involved in metabolism and transport processes. As more and more new classes of riboswitches are being discovered, it is important to understand the patterns of their intra and inter genomic distribution. Understanding such patterns will enable us to better understand the evolutionary history of these genetic regulatory elements. However, a complete picture of the distribution pattern of riboswitches will emerge only after accurate identification of riboswitches across genomes. We believe that the riboswitch detection method developed in this paper will aid in that process. The significant advantage in terms of speed, of our pHMM-based approach over the method based on CM allows us to scan entire databases (rather than 5'UTRs only) in a relatively short period of time in order to accurately identify riboswitch candidates. PMID- 19814812 TI - Text mining and manual curation of chemical-gene-disease networks for the comparative toxicogenomics database (CTD). AB - BACKGROUND: The Comparative Toxicogenomics Database (CTD) is a publicly available resource that promotes understanding about the etiology of environmental diseases. It provides manually curated chemical-gene/protein interactions and chemical- and gene-disease relationships from the peer-reviewed, published literature. The goals of the research reported here were to establish a baseline analysis of current CTD curation, develop a text-mining prototype from readily available open source components, and evaluate its potential value in augmenting curation efficiency and increasing data coverage. RESULTS: Prototype text-mining applications were developed and evaluated using a CTD data set consisting of manually curated molecular interactions and relationships from 1,600 documents. Preliminary results indicated that the prototype found 80% of the gene, chemical, and disease terms appearing in curated interactions. These terms were used to re rank documents for curation, resulting in increases in mean average precision (63% for the baseline vs. 73% for a rule-based re-ranking), and in the correlation coefficient of rank vs. number of curatable interactions per document (baseline 0.14 vs. 0.38 for the rule-based re-ranking). CONCLUSION: This text mining project is unique in its integration of existing tools into a single workflow with direct application to CTD. We performed a baseline assessment of the inter-curator consistency and coverage in CTD, which allowed us to measure the potential of these integrated tools to improve prioritization of journal articles for manual curation. Our study presents a feasible and cost-effective approach for developing a text mining solution to enhance manual curation throughput and efficiency. PMID- 19814813 TI - Phylogeny and evolutionary history of Leymus (Triticeae; Poaceae) based on a single-copy nuclear gene encoding plastid acetyl-CoA carboxylase. AB - BACKGROUND: Single- and low- copy genes are less likely subject to concerted evolution, thus making themselves ideal tools for studying the origin and evolution of polyploid taxa. Leymus is a polyploid genus with a diverse array of morphology, ecology and distribution in Triticeae. The genomic constitution of Leymus was assigned as NsXm, where Ns was presumed to be originated from Psathyrostachys, while Xm represented a genome of unknown origin. In addition, little is known about the evolutionary history of Leymus. Here, we investigate the phylogenetic relationship, genome donor, and evolutionary history of Leymus based on a single-copy nuclear Acc1 gene. RESULTS: Two homoeologues of the Acc1 gene were isolated from nearly all the sampled Leymus species using allele specific primer and were analyzed with those from 35 diploid taxa representing 18 basic genomes in Triticeae. Sequence diversity patterns and genealogical analysis suggested that (1) Leymus is closely related to Psathyrostachys, Agropyron, and Eremopyrum; (2) Psathyrostachys juncea is an ancestral Ns-genome donor of Leymus species; (3) the Xm genome in Leymus may be originated from an ancestral lineage of Agropyron and Eremopyrum triticeum; (4) the Acc1 sequences of Leymus species from the Qinghai-Tibetan plateau are evolutionarily distinct; (5) North America Leymus species might originate from colonization via the Bering land bridge; (6) Leymus originated about 11-12MYA in Eurasia, and adaptive radiation might have occurred in Leymus during the period of 3.7-4.3 MYA and 1.7-2.1 MYA. CONCLUSION: Leymus species have allopolyploid origin. It is hypothesized that the adaptive radiation of Leymus species might have been triggered by the recent upliftings of the Qinghai-Tibetan plateau and subsequent climatic oscillations. Adaptive radiation may have promoted the rapid speciation, as well as the fixation of unique morphological characters in Leymus. Our results shed new light on our understanding of the origin of Xm genome, the polyploidization events and evolutionary history of Leymus that could account for the rich diversity and ecological adaptation of Leymus species. PMID- 19814814 TI - Antiretroviral treatment outcomes from a nurse-driven, community-supported HIV/AIDS treatment programme in rural Lesotho: observational cohort assessment at two years. AB - INTRODUCTION: Lesotho has the third highest HIV prevalence in the world (an adult prevalence of 23.2%). Despite a lack of resources for health, the country has implemented state-of-the-art antiretroviral treatment guidelines, including early initiation of treatment (<350 cells/mm3), tenofovir in first line, and nurse initiated and managed HIV care, including antiretroviral therapy (ART), at primary health care level. PROGRAMME APPROACH: We describe two-year outcomes of a decentralized HIV/AIDS care programme run by Doctors Without Borders/Medecins Sans Frontieres, the Ministry of Health and Social Welfare, and the Christian Health Association of Lesotho in Scott catchment area, a rural health zone covering 14 clinics and one district hospital. Outcome data are described through a retrospective cohort analysis of adults and children initiated on ART between 2006 and 2008. DISCUSSION AND EVALUATION: Overall, 13,243 people have been enrolled in HIV care (5% children), and 5376 initiated on ART (6.5% children), 80% at primary care level. Between 2006 and 2008, annual enrolment more than doubled for adults and children, with no major external increase in human resources. The proportion of adults arriving sick (CD4 <50 cells/mm3) decreased from 22.2% in 2006 to 11.9% in 2008. Twelve-month outcomes are satisfactory in terms of mortality (11% for adults; 9% for children) and loss to follow up (8.8%). At 12 months, 80% of adults and 89% of children were alive and in care, meaning they were still taking their treatment; at 24 months, 77% of adults remained in care. CONCLUSION: Despite major resource constraints, Lesotho is comparing favourably with its better resourced neighbour, using the latest international ART recommendations. The successful two-year outcomes are further evidence that HIV/AIDS care and treatment can be provided effectively at the primary care level. The programme highlights how improving HIV care strengthened the primary health care system, and validates several critical areas for task shifting that are being considered by other countries in the region, including nurse-driven ART for adults and children, and lay counsellor-supported testing and counselling, adherence and case management. PMID- 19814815 TI - A first generation BAC-based physical map of the rainbow trout genome. AB - BACKGROUND: Rainbow trout (Oncorhynchus mykiss) are the most-widely cultivated cold freshwater fish in the world and an important model species for many research areas. Coupling great interest in this species as a research model with the need for genetic improvement of aquaculture production efficiency traits justifies the continued development of genomics research resources. Many quantitative trait loci (QTL) have been identified for production and life history traits in rainbow trout. A bacterial artificial chromosome (BAC) physical map is needed to facilitate fine mapping of QTL and the selection of positional candidate genes for incorporation in marker-assisted selection (MAS) for improving rainbow trout aquaculture production. This resource will also facilitate efforts to obtain and assemble a whole-genome reference sequence for this species. RESULTS: The physical map was constructed from DNA fingerprinting of 192,096 BAC clones using the 4-color high-information content fingerprinting (HICF) method. The clones were assembled into physical map contigs using the finger-printing contig (FPC) program. The map is composed of 4,173 contigs and 9,379 singletons. The total number of unique fingerprinting fragments (consensus bands) in contigs is 1,185,157, which corresponds to an estimated physical length of 2.0 Gb. The map assembly was validated by 1) comparison with probe hybridization results and agarose gel fingerprinting contigs; and 2) anchoring large contigs to the microsatellite-based genetic linkage map. CONCLUSION: The production and validation of the first BAC physical map of the rainbow trout genome is described in this paper. We are currently integrating this map with the NCCCWA genetic map using more than 200 microsatellites isolated from BAC end sequences and by identifying BACs that harbor more than 300 previously mapped markers. The availability of an integrated physical and genetic map will enable detailed comparative genome analyses, fine mapping of QTL, positional cloning, selection of positional candidate genes for economically important traits and the incorporation of MAS into rainbow trout breeding programs. PMID- 19814816 TI - The word landscape of the non-coding segments of the Arabidopsis thaliana genome. AB - BACKGROUND: Genome sequences can be conceptualized as arrangements of motifs or words. The frequencies and positional distributions of these words within particular non-coding genomic segments provide important insights into how the words function in processes such as mRNA stability and regulation of gene expression. RESULTS: Using an enumerative word discovery approach, we investigated the frequencies and positional distributions of all 65,536 different 8-letter words in the genome of Arabidopsis thaliana. Focusing on promoter regions, introns, and 3' and 5' untranslated regions (3'UTRs and 5'UTRs), we compared word frequencies in these segments to genome-wide frequencies. The statistically interesting words in each segment were clustered with similar words to generate motif logos. We investigated whether words were clustered at particular locations or were distributed randomly within each genomic segment, and we classified the words using gene expression information from public repositories. Finally, we investigated whether particular sets of words appeared together more frequently than others. CONCLUSION: Our studies provide a detailed view of the word composition of several segments of the non-coding portion of the Arabidopsis genome. Each segment contains a unique word-based signature. The respective signatures consist of the sets of enriched words, 'unwords', and word pairs within a segment, as well as the preferential locations and functional classifications for the signature words. Additionally, the positional distributions of enriched words within the segments highlight possible functional elements, and the co-associations of words in promoter regions likely represent the formation of higher order regulatory modules. This work is an important step toward fully cataloguing the functional elements of the Arabidopsis genome. PMID- 19814817 TI - Investigating the robustness of the classical enzyme kinetic equations in small intracellular compartments. AB - BACKGROUND: Classical descriptions of enzyme kinetics ignore the physical nature of the intracellular environment. Main implicit assumptions behind such approaches are that reactions occur in compartment volumes which are large enough so that molecular discreteness can be ignored and that molecular transport occurs via diffusion. Though these conditions are frequently met in laboratory conditions, they are not characteristic of the intracellular environment, which is compartmentalized at the micron and submicron scales and in which active means of transport play a significant role. RESULTS: Starting from a master equation description of enzyme reaction kinetics and assuming metabolic steady-state conditions, we derive novel mesoscopic rate equations which take into account (i) the intrinsic molecular noise due to the low copy number of molecules in intracellular compartments (ii) the physical nature of the substrate transport process, i.e. diffusion or vesicle-mediated transport. These equations replace the conventional macroscopic and deterministic equations in the context of intracellular kinetics. The latter are recovered in the limit of infinite compartment volumes. We find that deviations from the predictions of classical kinetics are pronounced (hundreds of percent in the estimate for the reaction velocity) for enzyme reactions occurring in compartments which are smaller than approximately 200 nm, for the case of substrate transport to the compartment being mediated principally by vesicle or granule transport and in the presence of competitive enzyme inhibitors. CONCLUSION: The derived mesoscopic rate equations describe subcellular enzyme reaction kinetics, taking into account, for the first time, the simultaneous influence of both intrinsic noise and the mode of transport. They clearly show the range of applicability of the conventional deterministic equation models, namely intracellular conditions compatible with diffusive transport and simple enzyme mechanisms in several hundred nanometre sized compartments. An active transport mechanism coupled with large intrinsic noise in enzyme concentrations is shown to lead to huge deviations from the predictions of deterministic models. This has implications for the common approach of modeling large intracellular reaction networks using ordinary differential equations and also for the calculation of the effective dosage of competitive inhibitor drugs. PMID- 19814818 TI - Employment in the Ecuadorian cut-flower industry and the risk of spontaneous abortion. AB - BACKGROUND: Research on the potentially adverse effects of occupational pesticide exposure on risk of spontaneous abortion (SAB) is limited, particularly among female agricultural workers residing in developing countries. METHODS: Reproductive histories were obtained from 217 Ecuadorian mothers participating in a study focusing on occupational pesticide exposure and children's neurobehavioral development. Only women with 2+ pregnancies were included in this study (n = 153). Gravidity, parity and frequency of SAB were compared between women with and without a history of working in the cut-flower industry in the previous 6 years. Logistic regression analysis was conducted to assess the relation between SAB and employment in the flower industry adjusting for maternal age. RESULTS: In comparison to women not working in the flower industry, women working in the flower industry were significantly younger (27 versus 32 years) and of lower gravidity (3.3 versus 4.5) and reported more pregnancy losses. A 2.6 (95% CI: 1.03-6.7) fold increase in the odds of pregnancy loss among exposed women was observed after adjusting for age. Odds of reporting an SAB increased with duration of flower employment, increasing to 3.4 (95% CI: 1.3, 8.8) among women working 4 to 6 years in the flower industry compared to women who did not work in the flower industry. CONCLUSION: This exploratory analysis suggests a potential adverse association between employment in the cut-flower industry and SAB. Study limitations include the absence of a temporal relation between exposure and SAB, no quantification of specific pesticides, and residual confounding such as physical stressors (i.e., standing). Considering that approximately half of the Ecuadorian flower laborers are women, our results emphasize the need for an evaluating the reproductive health effects of employment in the flower industry on reproductive health in this population. PMID- 19814819 TI - Melanocortin peptides inhibit urate crystal-induced activation of phagocytic cells. AB - INTRODUCTION: The melanocortin peptides have marked anti-inflammatory potential, primarily through inhibition of proinflammatory cytokine production and action on phagocytic cell functions. Gout is an acute form of arthritis caused by the deposition of urate crystals, in which phagocytic cells and cytokines play a major pathogenic role. We examined whether alpha-melanocyte-stimulating hormone (alpha-MSH) and its synthetic derivative (CKPV)2 influence urate crystal-induced monocyte (Mo) activation and neutrophil responses in vitro. METHODS: Purified Mos were stimulated with monosodium urate (MSU) crystals in the presence or absence of melanocortin peptides. The supernatants were tested for their ability to induce neutrophil activation in terms of chemotaxis, production of reactive oxygen intermediates (ROIs), and membrane expression of CD11b, Toll-like receptor 2 (TLR2) and TLR4. The proinflammatory cytokines interleukin (IL)-1beta, IL-8, and tumor necrosis factor-alpha (TNF-alpha) and caspase-1 were determined in the cell-free supernatants. In parallel experiments, purified neutrophils were preincubated overnight with or without melanocortin peptides before the functional assays. RESULTS: The supernatants from MSU crystal-stimulated Mos exerted chemoattractant and priming activity on neutrophils, estimated as ROI production and CD11b membrane expression. The supernatants of Mos stimulated with MSU in the presence of melanocortin peptides had less chemoattractant activity for neutrophils and less ability to prime neutrophils for CD11b membrane expression and oxidative burst. MSU crystal-stimulated Mos produced significant levels of IL-1beta, IL-8, TNF-alpha, and caspase-1. The concentrations of proinflammatory cytokines, but not of caspase-1, were reduced in the supernatants from Mos stimulated by MSU crystals in the presence of melanocortin peptides. Overnight incubation of neutrophils with the peptides significantly inhibited their ability to migrate toward chemotactic supernatants and their capacity to be primed in terms of ROI production. CONCLUSIONS: Alpha-MSH and (CKPV)2 have a dual effect on MSU crystal-induced inflammation, inhibiting the Mos' ability to produce neutrophil chemoattractants and activating compounds and preventing the neutrophil responses to these proinflammatory substances. These findings reinforce previous observations on the potential role of alpha-MSH and related peptides as a new class of drugs for treatment of inflammatory arthritis. PMID- 19814820 TI - Immunological considerations of modern animal models of malignant primary brain tumors. AB - Recent advances in animal models of glioma have facilitated a better understanding of biological mechanisms underlying gliomagenesis and glioma progression. The limitations of existing therapy, including surgery, chemotherapy, and radiotherapy, have prompted numerous investigators to search for new therapeutic approaches to improve quantity and quality of survival from these aggressive lesions. One of these approaches involves triggering a tumor specific immune response. However, a difficulty in this approach is the the scarcity of animal models of primary CNS neoplasms which faithfully recapitulate these tumors and their interaction with the host's immune system. In this article, we review the existing methods utilized to date for modeling gliomas in rodents, with a focus on the known as well as potential immunological aspects of these models. As this review demonstrates, many of these models have inherent immune system limitations, and the impact of these limitations on studies on the influence of pre-clinical therapeutics testing warrants further attention. PMID- 19814821 TI - Vitamin C supplement use may protect against gallstones: an observational study on a randomly selected population. AB - BACKGROUND: Animal experiments have shown a protective effect of vitamin C on the formation of gallstones. Few data in humans suggest an association between reduced vitamin C intake and increased prevalence of gallstone disease. The aim of this study was to assess the possible association of regular vitamin C supplementation with gallstone prevalence. METHODS: An observational, population based study of 2129 subjects aged 18-65 years randomly selected from the general population in southern Germany was conducted. Abdominal ultrasound examination, completion of a standardized questionnaire, compilation of anthropometric data and blood tests were used. Data were collected in November and December 2002. Data analysis was conducted between December 2005 and January 2006. RESULTS: Prevalence of gallstones in the study population was 7.8% (167/2129). Subjects reporting vitamin C supplementation showed a prevalence of 4.7% (11/232), whereas in subjects not reporting regular vitamin C supplementation, the prevalence was 8.2% (156/1897). Female gender, hereditary predisposition, increasing age and body-mass index (BMI) were associated with increased prevalence of gallstones. Logistic regression with backward elimination adjusted for these factors showed reduced gallstone prevalence for vitamin C supplementation (odds ratio, OR 0.34; 95% confidence interval, CI 0.14 to 0.81; P = 0.01), increased physical activity (OR 0.62; 95% CI, 0.42 to 0.94; P = 0.02), and higher total cholesterol (OR 0.65; 95% CI, 0.52 to 0.79; P < 0.001). CONCLUSION: Regular vitamin C supplementation and, to a lesser extent, increased physical activity and total cholesterol levels are associated with a reduced prevalence of gallstones. Regular vitamin C supplementation might exert a protective effect on the development of gallstones. PMID- 19814822 TI - Is the intraosseous access route fast and efficacious compared to conventional central venous catheterization in adult patients under resuscitation in the emergency department? A prospective observational pilot study. AB - BACKGROUND: For patients' safety reasons, current American Heart Association and European Resuscitation Council guidelines recommend intraosseous (IO) vascular access as an alternative in cases of emergency, if prompt venous catheterization is impossible. The purpose of this study was to compare the IO access as a bridging procedure versus central venous catheterization (CVC) for in-hospital adult emergency patients under resuscitation with impossible peripheral intravenous (IV) access. We hypothesised, that CVC is faster and more efficacious compared to IO access. METHODS: A prospective observational study comparing success rates and procedure times of IO access (EZ-IO, Vidacare Corporation) versus CVC in adult (>/=18 years of age) patients under trauma and medical resuscitation admitted to our emergency department with impossible peripheral IV catheterization was conducted. Procedure time was defined from preparation and insertion of vascular access type until first drug or infusion solution administration. Success rate on first attempt and procedure time for each access route was evaluated and statistically tested. RESULTS: Ten consecutive adult patients under resuscitation, each receiving IO access and CVC, were analyzed. IO access was performed with 10 tibial or humeral insertions, CVC in 10 internal jugular or subclavian veins. The success rate on first attempt was 90% for IO insertion versus 60% for CVC. Mean procedure time was significantly lower for IO cannulation (2.3 min +/- 0.8) compared to CVC (9.9 min +/- 3.7) (p < 0.001). As for complications, failure of IO access was observed in one patient, while two or more attempts of CVC were necessary in four patients. No other relevant complications, like infection, bleeding or pneumothorax were observed. CONCLUSION: Preliminary data demonstrate that IO access is a reliable bridging method to gain vascular access for in-hospital adult emergency patients under trauma or medical resuscitation with impossible peripheral IV access. Furthermore, IO cannulation requires significantly less time to enable administration of drugs or infusion solutions compared to CVC. Because CVC was slower and less efficacious, IO access may improve the safety of adult patients under resuscitation in the emergency department. PMID- 19814823 TI - Homology-based annotation of non-coding RNAs in the genomes of Schistosoma mansoni and Schistosoma japonicum. AB - BACKGROUND: Schistosomes are trematode parasites of the phylum Platyhelminthes. They are considered the most important of the human helminth parasites in terms of morbidity and mortality. Draft genome sequences are now available for Schistosoma mansoni and Schistosoma japonicum. Non-coding RNA (ncRNA) plays a crucial role in gene expression regulation, cellular function and defense, homeostasis, and pathogenesis. The genome-wide annotation of ncRNAs is a non trivial task unless well-annotated genomes of closely related species are already available. RESULTS: A homology search for structured ncRNA in the genome of S. mansoni resulted in 23 types of ncRNAs with conserved primary and secondary structure. Among these, we identified rRNA, snRNA, SL RNA, SRP, tRNAs and RNase P, and also possibly MRP and 7SK RNAs. In addition, we confirmed five miRNAs that have recently been reported in S. japonicum and found two additional homologs of known miRNAs. The tRNA complement of S. mansoni is comparable to that of the free living planarian Schmidtea mediterranea, although for some amino acids differences of more than a factor of two are observed: Leu, Ser, and His are overrepresented, while Cys, Meth, and Ile are underrepresented in S. mansoni. On the other hand, the number of tRNAs in the genome of S. japonicum is reduced by more than a factor of four. Both schistosomes have a complete set of minor spliceosomal snRNAs. Several ncRNAs that are expected to exist in the S. mansoni genome were not found, among them the telomerase RNA, vault RNAs, and Y RNAs. CONCLUSION: The ncRNA sequences and structures presented here represent the most complete dataset of ncRNA from any lophotrochozoan reported so far. This data set provides an important reference for further analysis of the genomes of schistosomes and indeed eukaryotic genomes at large. PMID- 19814824 TI - Conformational changes and loose packing promote E. coli Tryptophanase cold lability. AB - BACKGROUND: Oligomeric enzymes can undergo a reversible loss of activity at low temperatures. One such enzyme is tryptophanase (Trpase) from Escherichia coli. Trpase is a pyridoxal phosphate (PLP)-dependent tetrameric enzyme with a Mw of 210 kD. PLP is covalently bound through an enamine bond to Lys270 at the active site. The incubation of holo E. coli Trpases at 2 degrees C for 20 h results in breaking this enamine bond and PLP release, as well as a reversible loss of activity and dissociation into dimers. This sequence of events is termed cold lability and its understanding bears relevance to protein stability and shelf life. RESULTS: We studied the reversible cold lability of E. coli Trpase and its Y74F, C298S and W330F mutants. In contrast to the holo E. coli Trpase all apo forms of Trpase dissociated into dimers already at 25 degrees C and even further upon cooling to 2 degrees C. The crystal structures of the two mutants, Y74F and C298S in their apo form were determined at 1.9A resolution. These apo mutants were found in an open conformation compared to the closed conformation found for P. vulgaris in its holo form. This conformational change is further supported by a high pressure study. CONCLUSION: We suggest that cold lability of E. coli Trpases is primarily affected by PLP release. The enhanced loss of activity of the three mutants is presumably due to the reduced size of the side chain of the amino acids. This prevents the tight assembly of the active tetramer, making it more susceptible to the cold driven changes in hydrophobic interactions which facilitate PLP release. The hydrophobic interactions along the non catalytic interface overshadow the effect of point mutations and may account for the differences in the dissociation of E. coli Trpase to dimers and P. vulgaris Trpase to monomers. PMID- 19814825 TI - Human stool contains a previously unrecognized diversity of novel astroviruses. AB - Human astroviruses are a leading cause of gastrointestinal disease. Since their discovery in 1975, 8 closely related serotypes have been described in humans, and more recently, two new astrovirus species, astrovirus MLB1 and astrovirus VA1, were identified in diarrhea patients. In this study, we used consensus astrovirus primers targeting the RNA polymerase to define the diversity of astroviruses present in pediatric patients with diarrhea on two continents. From 416 stool specimens comprising two different cohorts from Vellore, India, 35 samples were positive. These positive samples were analyzed further by either sequencing of the approximately 400 bp amplicon generated by the consensus PCR or by performing additional RT-PCR specific for individual astroviruses. 19 samples contained the classic human astrovirus serotypes 1-8 while 7 samples were positive for the recently described astrovirus MLB1. Strikingly, from samples that were positive in the consensus PCR screen but negative in the specific PCR assays, five samples contained sequences that were highly divergent from all previously described astroviruses. Sequence analysis suggested that three novel astroviruses, tentatively named astroviruses VA2, MLB2 and VA3, were present in these five patient specimens (AstV-VA2 in 2 patients, AstV-MLB2 in 2 patients and AstV-VA3 in one patient). Using the same RT-PCR screening strategy, 13 samples out of 466 tested stool specimens collected in St. Louis, USA were positive. Nine samples were positive for the classic human astroviruses. One sample was positive for AstV-VA2, and 3 samples were positive for AstV-MLB2 demonstrating that these two viruses are globally widespread. Collectively, these findings underscore the tremendous diversity of astroviruses present in fecal specimens from diarrhea patients. Given that a significant fraction of diarrhea etiologies is currently unknown, it is plausible that these or other yet unrecognized astroviruses may be responsible for at least part of the undiagnosed cases. PMID- 19814826 TI - Information management to enable personalized medicine: stakeholder roles in building clinical decision support. AB - BACKGROUND: Advances in technology and the scientific understanding of disease processes are presenting new opportunities to improve health through individualized approaches to patient management referred to as personalized medicine. Future health care strategies that deploy genomic technologies and molecular therapies will bring opportunities to prevent, predict, and pre-empt disease processes but will be dependent on knowledge management capabilities for health care providers that are not currently available. A key cornerstone to the potential application of this knowledge will be effective use of electronic health records. In particular, appropriate clinical use of genomic test results and molecularly-targeted therapies present important challenges in patient management that can be effectively addressed using electronic clinical decision support technologies. DISCUSSION: Approaches to shaping future health information needs for personalized medicine were undertaken by a work group of the American Health Information Community. A needs assessment for clinical decision support in electronic health record systems to support personalized medical practices was conducted to guide health future development activities. Further, a suggested action plan was developed for government, researchers and research institutions, developers of electronic information tools (including clinical guidelines, and quality measures), and standards development organizations to meet the needs for personalized approaches to medical practice. In this article, we focus these activities on stakeholder organizations as an operational framework to help identify and coordinate needs and opportunities for clinical decision support tools to enable personalized medicine. SUMMARY: This perspective addresses conceptual approaches that can be undertaken to develop and apply clinical decision support in electronic health record systems to achieve personalized medical care. In addition, to represent meaningful benefits to personalized decision-making, a comparison of current and future applications of clinical decision support to enable individualized medical treatment plans is presented. If clinical decision support tools are to impact outcomes in a clear and positive manner, their development and deployment must therefore consider the needs of the providers, including specific practice needs, information workflow, and practice environment. PMID- 19814827 TI - The self-assessment scale of cognitive complaints in schizophrenia: a validation study in Tunisian population. AB - BACKGROUND: Despite a huge well-documented literature on cognitive deficits in schizophrenia, little is known about the own perception of patients regarding their cognitive functioning. The purpose of our study was to create a scale to collect subjective cognitive complaints of patients suffering from schizophrenia with Tunisian Arabic dialect as mother tongue and to proceed to a validation study of this scale. METHODS: The authors constructed the Self-Assessment Scale of Cognitive Complaints in Schizophrenia (SASCCS) based on a questionnaire covering five cognitive domains which are the most frequently reported in the literature to be impaired in schizophrenia. The scale consisted of 21 likert-type questions dealing with memory, attention, executive functions, language and praxia. In a second time, the authors proceeded to the study of psychometric qualities of the scale among 105 patients suffering from schizophrenia spectrum disorders (based on DSM- IV criteria). Patients were evaluated using the Positive and Negative Syndrome Scale (PANSS), the Global Assessment Functioning Scale (GAF scale) and the Calgary Depression Scale (CDS). RESULTS: The scale's reliability was proven to be good through Cronbach alpha coefficient equal to 0.85 and showing its good internal consistency. The intra-class correlation coefficient at 11 weeks was equal to 0.77 suggesting a good stability over time. Principal component analysis with Oblimin rotation was performed and yielded to six factors accounting for 58.28% of the total variance of the scale. CONCLUSION: Given the good psychometric properties that have been revealed in this study, the SASCCS seems to be reliable to measure schizophrenic patients' perception of their own cognitive impairment. This kind of evaluation can't substitute for objective measures of cognitive performances in schizophrenia. The purpose of such an evaluation is to permit to the patient to express his own well-being and satisfaction of quality of life. PMID- 19814828 TI - Pichinde virus induces microvascular endothelial cell permeability through the production of nitric oxide. AB - This report is the first to demonstrate infection of human endothelial cells by Pichinde virus (PIC). PIC infection induces an upregulation of the inducible nitric oxide synthase gene; as well as an increase in detectable nitric oxide (NO). PIC induces an increase in permeability in endothelial cell monolayers which can be abrogated at all measured timepoints with the addition of a nitric oxide synthase inhibitor, indicating a role for NO in the alteration of endothelial barrier function. Because NO has shown antiviral activity against some viruses, viral titer was measured after addition of the NO synthase inhibitor and found to have no effect in altering virus load in infected EC. The NO synthase inhibition also has no effect on levels of activated caspases induced by PIC infection. Taken together, these data indicate NO production induced by Pichinde virus infection has a pathogenic effect on endothelial cell monolayer permeability. PMID- 19814829 TI - The immediate effect of individual manipulation techniques on pulmonary function measures in persons with chronic obstructive pulmonary disease. AB - BACKGROUND: The use of manipulation has long been advocated in the treatment of chronic obstructive pulmonary disease (COPD), but few randomized controlled clinical trials have measured the effect of manipulation on pulmonary function. In addition, the effects of individual manipulative techniques on the pulmonary system are poorly understood. Therefore, the purpose of this study was to determine the immediate effects of four osteopathic techniques on pulmonary function measures in persons with COPD relative to a minimal-touch control protocol. METHODS: Persons with COPD aged 50 and over were recruited for the study. Subjects received five, single-technique treatment sessions: minimal-touch control, thoracic lymphatic pump (TLP) with activation, TLP without activation, rib raising, and myofascial release. There was a 4-week washout period between sessions. Protocols were given in random order until all five techniques had been administered. Pulmonary function measures were obtained at baseline and 30 minutes posttreatment. For the actual pulmonary function measures and percent predicted values, Wilcoxon signed rank tests were used to test within-technique changes from baseline. For the percent change from baseline, Friedman tests were used to test for between-technique differences. RESULTS: Twenty-five subjects were enrolled in the study. All four tested osteopathic techniques were associated with adverse posttreatment changes in pulmonary function measures; however, different techniques changed different measures. TLP with activation increased posttreatment residual volume compared to baseline, while TLP without activation did not. Side effects were mild, mostly posttreatment chest wall soreness. Surprisingly, the majority of subjects believed they could breathe better after receiving osteopathic manipulation. CONCLUSION: In persons with COPD, TLP with activation, TLP without activation, rib raising, and myofascial release mildly worsened pulmonary function measures immediately posttreatment relative to baseline measurements. The activation component of the TLP technique appears to increase posttreatment residual volume. Despite adverse changes in pulmonary function measures, persons with COPD subjectively reported they benefited from osteopathic manipulation. PMID- 19814830 TI - Microbicides development programme: engaging the community in the standard of care debate in a vaginal microbicide trial in Mwanza, Tanzania. AB - BACKGROUND: HIV prevention research in resource-limited countries is associated with a variety of ethical dilemmas. Key amongst these is the question of what constitutes an appropriate standard of health care (SoC) for participants in HIV prevention trials. This paper describes a community-focused approach to develop a locally-appropriate SoC in the context of a phase III vaginal microbicide trial in Mwanza City, northwest Tanzania. METHODS: A mobile community-based sexual and reproductive health service for women working as informal food vendors or in traditional and modern bars, restaurants, hotels and guesthouses has been established in 10 city wards. Wards were divided into geographical clusters and community representatives elected at cluster and ward level. A city-level Community Advisory Committee (CAC) with representatives from each ward has been established. Workshops and community meetings at ward and city-level have explored project-related concerns using tools adapted from participatory learning and action techniques e.g. chapati diagrams, pair-wise ranking. Secondary stakeholders representing local public-sector and non-governmental health and social care providers have formed a trial Stakeholders' Advisory Group (SAG), which includes two CAC representatives. RESULTS: Key recommendations from participatory community workshops, CAC and SAG meetings conducted in the first year of the trial relate to the quality and range of clinic services provided at study clinics as well as broader standard of care issues. Recommendations have included streamlining clinic services to reduce waiting times, expanding services to include the children and spouses of participants and providing care for common local conditions such as malaria. Participants, community representatives and stakeholders felt there was an ethical obligation to ensure effective access to antiretroviral drugs and to provide supportive community-based care for women identified as HIV positive during the trial. This obligation includes ensuring sustainable, post-trial access to these services. Post-trial access to an effective vaginal microbicide was also felt to be a moral imperative. CONCLUSION: Participatory methodologies enabled effective partnerships between researchers, participant representatives and community stakeholders to be developed and facilitated local dialogue and consensus on what constitutes a locally appropriate standard of care in the context of a vaginal microbicide trial in this setting. TRIAL REGISTRATION: Current Controlled Trials ISRCTN64716212. PMID- 19814831 TI - Health-promoting lifestyles of university students in mainland China. AB - BACKGROUND: Health-promoting lifestyles of adolescents are closely related to their current and subsequent health status. However, few studies in mainland China have examined health-promoting behaviors among university students, notwithstanding the dramatic development of higher education over the past two decades. Moreover, no study has applied a standardized scale to such an investigation. The adolescent health promotion (AHP) scale has been developed and is commonly used for measuring adolescent health-promoting lifestyles in Taiwan. The aim of this study is to determine the appropriateness of the AHP for use in mainland China. METHODS: A cross-sectional study was performed on a total of 420 undergraduates, who were randomly selected using a two-stage stratified sampling method in a university in Guangzhou city, mainland China. The simplified Chinese version of the AHP scale, comprising six dimensions (Nutrition behavior, Social support, Life-appreciation, Exercise behavior, Health-responsibility and Stress management), was used to measure health-promoting lifestyles among undergraduates. The reliability of the AHP scale was assessed using split-half reliability coefficients, intraclass correlation coefficients and Cronbach's alpha coefficient. Validity was assessed by factor analysis and correlation analysis. Factors associated with health-promoting lifestyles were identified using multiple linear regression. RESULTS: Cronbach's coefficients were greater than 0.7 in all dimensions of the AHP scale except for Nutrition behavior (0.684). Intraclass correlation coefficients ranged from 0.689 to 0.921. Split half reliability coefficients were higher than 0.7 in three AHP dimensions (Social support, Life-appreciation and Exercise behavior). Our results were generally in accordance with the theoretical construction of the AHP scale. The mean score for each of the six dimensions was lower than 70. Gender and grade were the factors primarily associated with health-promoting lifestyles among undergraduates. CONCLUSION: The AHP is a valid and reliable instrument for assessing health-promoting lifestyles of undergraduates in mainland China, which remain at a low level. Health behavior education taking account of gender and grade differences may also be applied. PMID- 19814832 TI - Community health needs assessment with precede-proceed model: a mixed methods study. AB - BACKGROUND: Community health services in China have developed over the last few decades. In order to use limited health resources more effectively, we conducted a community health needs assessment. This aimed to provide an understanding of the community's health problems and the range of potential factors affecting risk behaviours for the priority health problems. METHODS: We used the precede-proceed model for the needs assessment. Triangulation of data, methods and researchers were employed in data collection. RESULTS: Main findings include: cardiovascular diseases (CVDs) were identified as the priority health problems in the study communities; risk factors associated with CVDs included smoking, physical inactivity and unhealthy eating behaviours, particularly amongst male residents with low education level; factors negatively affecting behaviours were classified into predisposing factors (limited knowledge, beliefs and lack of perceived needs), enabling factors (limited access to health promotion activities, unawareness of health promotion, lack of work-site and school health promotion, absence of health promotion related policy) and reinforcing factors (culture). Policies and organization were not perfect; there were limited staff skilled in providing health promotion in the community. CONCLUSION: CVDs were identified by the communities as priority health problems. Future health programs should focus on smoking, physical inactivity and unhealthy eating behaviours. Behaviour change strategies should take predisposing factors, enabling factors and reinforcing factors into consideration. Policies, organization and human resource need strengthening. PMID- 19814833 TI - The impact of extended electrodiagnostic studies in ulnar neuropathy at the elbow. AB - BACKGROUND: This study aimed to explore the value of extended motor nerve conduction studies in patients with ulnar nerve entrapment at the elbow (UNE) in order to find the most sensitive and least time-consuming method. We wanted to evaluate the utility of examining both the sensory branch from the fifth finger and the dorsal branch of the ulnar nerve. Further we intended to study the clinical symptoms and findings, and a possible correlation between the neurophysiological findings and pain. METHODS: The study was prospective, and 127 UNE patients who were selected consecutively from the list of patients, had a clinical and electrodiagnostic examination. Data from the most symptomatic arm were analysed and compared to the department's reference limits. Student's t - test, chi-square tests and multiple regression models were used. Two-side p values < 0.05 were considered as significant. RESULTS: Ulnar paresthesias (96%) were more common than pain (60%). Reduced ulnar sensitivity (86%) and muscle strength (48%) were the most common clinical findings. Adding a third stimulation site in the elbow mid-sulcus for motor conduction velocity (MCV) to abductor digiti minimi (ADM) increased the electrodiagnostic sensitivity from 80% to 96%. Additional recording of ulnar MCV to the first dorsal interosseus muscle (FDI) increased the sensitivity from 96% to 98%. The ulnar fifth finger and dorsal branch sensory studies were abnormal in 39% and 30% of patients, respectively. Abnormal electromyography in FDI was found in 49% of the patients. Patients with and without pain had generally similar conduction velocity parameter means. CONCLUSION: We recommend three stimulation sites at the elbow for MCV to ADM. Recording from FDI is not routinely indicated. Sensory studies and electromyography do not contribute much to the sensitivity of the electrodiagnostic evaluation, but they are useful to document axonal degeneration. Most conduction parameters are unrelated to the presence of pain. PMID- 19814834 TI - Do neurooncological patients and their significant others agree on quality of life ratings? AB - INTRODUCTION: Patients suffering from brain tumours often experience a wide range of cognitive impairments that impair their ability to report on their quality of life and symptom burden. The use of proxy ratings by significant others may be a promising alternative to gain information for medical decision making or research purposes, if self-ratings are not obtainable. Our study investigated the agreement of quality of life and symptom ratings by the patient him/herself or by a significant other. METHODS: Patients with primary brain tumours were recruited at the neurooncological outpatient unit of Innsbruck Medical University. Quality of life self- and proxy-ratings were collected using the EORTC QLQ-C30 and its brain cancer module, the QLQ-BN20. RESULTS: Between May 2005 and August 2007, 42 pairs consisting of a patient and his/her significant other were included in the study. Most of the employed quality of life scales showed fairly good agreement between patient- and proxy-ratings (median correlation 0.46). This was especially true for Physical Functioning, Sleeping Disturbances, Appetite Loss, Constipation, Taste Alterations, Visual Disorders, Motor Dysfunction, Communication Deficits, Hair Loss, Itchy Skin, Motor Dysfunction and Hair Loss. Worse rater agreement was found for Social Functioning, Emotional Functioning, Cognitive Functioning, Fatigue, Pain, Dyspnoea and Seizures. CONCLUSION: The assessment of quality of life in brain cancer patients through ratings from their significant others seems to be a feasible strategy to gain information about certain aspects of patient's quality of life and symptom burden, if the patient is not able to provide information himself. PMID- 19814835 TI - Antioxidant-enriched enteral nutrition and immuno-inflammatory response after major gastrointestinal tract surgery. AB - Major surgery induces an immuno-inflammatory response accompanied by oxidative stress that may impair cellular function and delay recovery. The objective of the study was to investigate the effect of an enteral supplement, containing glutamine and antioxidants, on circulating levels of immuno-inflammatory markers after major gastrointestinal tract surgery. Patients (n 21) undergoing major gastrointestinal tract surgery were randomised in a single-centre, open-label study. The effects on circulating levels of immuno-inflammatory markers were determined on the day before surgery and on days 1, 3, 5 and 7 after surgery. Major gastrointestinal surgery increased IL-6, TNF receptor 55/60 (TNF-R55) and C reactive protein (CRP). Surgery reduced human leucocyte antigen-DR (HLA-DR) expression on monocytes. CRP decrease was more pronounced in the first 7 d in the treatment group compared with the control group. In the treatment group, from the moment Module AOX was administered on day 1 after surgery, TNF receptor 75/80 (TNF-R75) level decreased until the third post-operative day and then stabilised, whereas in the control group the TNF-R75 level continued to increase. The results of the present pilot study suggest that enteral nutrition enriched with glutamine and antioxidants possibly moderates the immuno-inflammatory response (CRP, TNF R75) after surgery. PMID- 19814836 TI - The probiotic Lactobacillus acidophilus reduces cholesterol absorption through the down-regulation of Niemann-Pick C1-like 1 in Caco-2 cells. AB - Elevated blood cholesterol is an important risk factor associated with atherosclerosis and CHD. The search for mediators that fine tune cholesterol homeostasis has recognised probiotics as being potentially beneficial. Here, we present data describing bacterial regulation of Niemann-Pick C1-like 1 (NPC1L1), which, when weakly expressed, results in a marked reduction in intestinal absorption of cholesterol. The probiotic Lactobacillus acidophilus ATCC 4356 reduced NPC1L1 gene expression and inhibited the cellular uptake of micellar cholesterol in Caco-2 cells. Soluble effector molecules secreted by ATCC 4356 were shown to be responsible for the decrease in NPC1L1. Furthermore, ATCC 4356 mediated this effect partly through the liver X receptors (LXR). The role of NPC1L1 and the LXR in cholesterol metabolism underscores the basis for the use of probiotics, such as ATCC 4356, in managing hypercholesterolaemia. PMID- 19814837 TI - Trends in food intake in French children from 1999 to 2007: results from the INCA (etude Individuelle Nationale des Consommations Alimentaires) dietary surveys. AB - The objectives of the present study were to assess the intake of different food groups in French children aged 3-17 years (n 1455), and to analyse trends since a dietary survey undertaken 8 years ago. Dietary intake was evaluated using data from the 2006-7 cross-sectional INCA2 national dietary survey (etude Individuelle Nationale sur les Consommations Alimentaires), based on a 7 d food record. Dietary intake (percentage of subjects consuming the food group and amount eaten) was assessed for thirty-nine food categories. We observed variations in food consumption by age, sex, North-South regional gradient, seasonal period and educational level of the responding parent. Trends in dietary intake between 1999 and 2007 were determined by comparing the INCA1 (n 1126) and the INCA2 surveys. Both surveys had been carried out using the same methodology. The findings showed a decrease in energy intake in children aged 3-14 years, due to a reduction in the consumption of foods of animal origin and sweetened products. In adolescents aged 15-17 years, energy intake remained rather stable; during this 8-year period, the consumption of meat decreased, whereas the consumption of savoury snacks such as sandwiches and hamburgers significantly increased. These trends occurred during a time of growing concern about overweight and the associated co morbidities in France. A number of public health measures were implemented over this period to improve dietary habits and physical activity patterns in children and adults. The periodic monitoring of dietary patterns through the INCA surveys is an essential part of the surveillance network in France. PMID- 19814838 TI - The association between dietary protein intake and bone mass accretion in pubertal girls with low calcium intakes. AB - To assess the association between protein intakes and bone mass accrual in girls, data were analysed for 757 pre-pubertal girls (mean age 10.1 years) in urban Beijing, China, who participated in a 5-year study including 2 years of milk supplementation (intervention groups only) and 3 years of follow-up study. At 0, 12, 24, 48 and 60 months from the baseline, bone mass of the proximal or distal forearm (PF or DF) and total body (TB) was measured with dual energy X-ray absorptiometry; dietary intakes were assessed by a 3-d food record (including two weekdays and one weekend day). Linear mixed models were used and continuous variables were logarithm transformed. The mean longitudinal Ca intake (432-675 mg/d on average) positively influenced bone mineral content (BMC) at TB, PF and DF after controlling for baseline bone mass and other possible confounders. However, negative associations were observed between protein intake (55.9-61.0 g/d on average) and BMC accrual at TB, PF or DF (beta = - 1.92, - 10.2 or - 4.82, respectively, P < 0.01) after adjustment. When protein intake was considered according to animal or plant food sources, protein from animal foods, particularly meat, had significant negative effects on BMC accrual at DF or PF after adjustment. It was concluded that higher protein intake, especially from animal foods, appeared to have a negative effect on bone mass accrual in Chinese pubertal girls with low Ca intakes. PMID- 19814839 TI - The concept of dementia: retain, reframe, rename or replace? AB - From antiquity the term "dementia" has denoted a state of severe acquired intellectual deterioration which significantly interferes with the fulfillment of personal, social or occupational roles, and makes the individual dependent on care and supervision by others. The medical concept of dementia refers to a pattern of cognitive and behavioral symptoms which typically arises from chronic and often progressive brain diseases. The quantitative expression of this pattern shows broad variability, and some patients fall within the boundaries of the concept whose intellectual and functional abilities are only mildly impaired. On the other hand, the concept currently has an unduly narrow qualitative bandwidth, because it is modeled after the subtype which occurs in Alzheimer's disease but does not represent a good fit for other important subtypes. In the authors' view, the concept of dementia should be retained despite its limitations, since it has an important role in directing the physician's attention to a certain group of underlying pathologies. This diagnostic role of the concept will remain important in primary care even if biological indicators for one or several etiologies will become part of the diagnostic routine in research units in the future. The medical construct has further value since it entitles patients to medical treatment, social assistance and legal protection. Although in our opinion the concept of dementia does not need to be replaced, upcoming revisions of the psychiatric classification systems will have to reframe it by emphasizing the heterogeneity of the psychopathological symptom pattern. In view of the increasing importance of early diagnosis and treatment, however, the term "dementia", which literally means "absence of mind", is no longer an appropriate and timely designation for the broad range of cognitive and behavioral limitations covered by the concept. It should be renamed, using a terminology which accommodates scientific advance and meets the requirements of medical communication while preserving the benefits for patients and their families. PMID- 19814840 TI - Assessing speech and communication impairments in cognitive disorders: an innovative development in a memory clinic. PMID- 19814841 TI - The accuracy of the Clock Drawing Test compared to that of standard screening tests for Alzheimer's disease: results from a study of Brazilian elderly with heterogeneous educational backgrounds. AB - BACKGROUND: Although the Clock Drawing Test (CDT) is the second most used test in the world for the screening of dementia, there is still debate over its sensitivity, specificity, application and interpretation in dementia diagnosis. This study has three main aims: to evaluate the sensitivity and specificity of the CDT in a sample composed of older adults with Alzheimer's disease (AD) and normal controls; to compare CDT accuracy to the that of the Mini-mental State Examination (MMSE) and the Cambridge Cognitive Examination (CAMCOG); and to test whether the association of the MMSE with the CDT leads to higher or comparable accuracy as that reported for the CAMCOG. METHODS: Cross-sectional assessment was carried out for 121 AD and 99 elderly controls with heterogeneous educational levels from a geriatric outpatient clinic who completed the Cambridge Examination for Mental Disorder of the Elderly (CAMDEX). The CDT was evaluated according to the Shulman, Mendez and Sunderland scales. RESULTS: The CDT showed high sensitivity and specificity. There were significant correlations between the CDT and the MMSE (0.700-0.730; p < 0.001) and between the CDT and the CAMCOG (0.753 0.779; p < 0.001). The combination of the CDT with the MMSE improved sensitivity and specificity (SE = 89.2-90%; SP = 71.7-79.8%). Subgroup analysis indicated that for elderly people with lower education, sensitivity and specificity were both adequate and high. CONCLUSIONS: The CDT is a robust screening test when compared with the MMSE or the CAMCOG, independent of the scale used for its interpretation. The combination with the MMSE improves its performance significantly, becoming equivalent to the CAMCOG. PMID- 19814842 TI - Predicting memory decline as a risk factor for Alzheimer's disease in older post menopausal women: quod erat demonstrandum? PMID- 19814843 TI - An effective approach to decrease antipsychotic and benzodiazepine use in nursing homes: the RedUSe project. AB - BACKGROUND: This study evaluated a multi-faceted, interdisciplinary intervention to reduce the use of benzodiazepines and antipsychotics in nursing homes - the "RedUSe" (Reducing Use of Sedatives) project. METHODS: The RedUSe project was a controlled trial conducted in 25 nursing homes in Tasmania, with 13 intervention and 12 control homes. A series of pharmacist-led strategies were provided to intervention homes including two medication audit and feedback cycles, educational sessions for staff and an interdisciplinary sedative review. Data on psychotropic drug use at each nursing home were collected utilizing a customized computer program at baseline, 12 and 26 weeks. The RedUSe project was registered as a controlled trial at the Australian New Zealand Clinical Trials Registry, registration number: ACTRN12608000221358. RESULTS: For each measure, an average of 1591 residents were audited. Over the six-month trial, there was a significant reduction in the percentage of intervention home residents regularly taking benzodiazepines (31.8% to 26.9%, p < 0.005) and antipsychotics (20.3% to 18.6%, p < 0.05), whereas control home psychotropic use did not alter significantly. For residents taking benzodiazepines and antipsychotics at baseline, there were significantly more dose reductions/cessations in intervention homes than in control homes (benzodiazepines: 39.6% vs. 17.6%, p < 0.0001; antipsychotics: 36.9% vs. 20.9%, p < 0.01). CONCLUSIONS: RedUSe led to a significant reduction in the proportion of residents in nursing homes taking benzodiazepines and antipsychotics, and a significant increase in the number of dose reductions of these agents. Our findings suggest that a multi-faceted program, coordinated through a community pharmacy, can offer an effective approach in reducing psychotropic use in nursing homes. PMID- 19814844 TI - Morphological effects and tegumental alterations induced by mefloquine on schistosomula and adult flukes of Schistosoma mansoni. AB - There is a pressing need to develop novel anti-schistosomal drugs, as current treatment relies largely on praziquantel (PZQ). To further strengthen current evidence of the anti-schistosomal properties of mefloquine (MQ), we studied the temporal effect of this compound in vitro and in vivo, and examined alterations on the tegumental surface of schistosomula and adults of S. mansoni by means of scanning electron microscopy (SEM). Schistosomula and adults were each incubated in vitro using MQ over a wide concentration range (1-100 microg/ml). In addition, mice infected with adult S. mansoni were treated with a single oral dose of 400 mg/kg MQ, and worms were recovered 24, 48, 72, 96 and 120 h following treatment. MQ showed a rapid onset of action on schistosomula in vitro; 100 and 75 microg/ml of MQ killed schistosomula immediately; the minimal lethal and effective concentrations of MQ on schistosomula after 1 h were 25 and 5 microg/ml, respectively. Adult worms incubated with 100 and 10 microg/ml of MQ were dead after 1 h and 24 h of incubation, respectively. A hepatic shift of adult schistosomes was observed in mice already 24 h after treatment, and 120 h following treatment >98% of all worms had translocated to the liver. SEM observations revealed extensive tegumental destruction, including blebbing, shrinking and sloughing, particularly following in vitro incubation and on the tegument of female worms. PMID- 19814845 TI - Control of schistosomiasis in sub-Saharan Africa: progress made, new opportunities and remaining challenges. AB - Several other journal supplements have documented progress made in the control of schistosomiasis in Egypt, China and Brazil, however, with more than 97% of the schistosome infections now estimated to occur in Africa, the relevance of this special issue in Parasitology cannot be overemphasized. In total, 18 articles are presented, inclusive of a lead-editorial from the WHO highlighting a seminal resolution at the 54th World Health Assembly in 2001 that advocated de-worming. Facilitated by a US$ 30 million grant from the Bill and Melinda Gates Foundation in 2002, the Schistosomiasis Control Initiative subsequently fostered implementation of large-scale schistosomiasis (and soil-transmitted helminthiasis) control programmes in six selected African countries. From 2005, CONTRAST, a European union-funded consortium, was formed to conduct multi disciplinary research pertaining to optimisation of schistosomiasis control. Progress made in schistosomiasis control across sub-Saharan Africa since the turn of the new millennium is reviewed, shedding light on the latest findings stemming from clinical, epidemiological, molecular and social sciences research, inclusive of public health interventions with monitoring and evaluation activities. New opportunities for integrating the control of schistosomiasis and other so-called neglected tropical diseases are highlighted, but more importantly, several opportune questions that arise from it frame the remaining challenges ahead for an enduring solution. PMID- 19814846 TI - Mesocestoides corti: morphological features and glycogen mobilization during in vitro differentiation from larva to adult worm. AB - Mesocestodes corti has the capacity to develop from the tetrathyridium (larva) stage to adult worm in vitro by trypsin and serum stimulation. Consequently, it has been used as an experimental model system for studying cestode development, host-parasite relationships and anthelmintic drugs. We describe morphological features in 5 different developmental stages of M. corti obtained in vitro, including larvae from the peritoneal cavity of infected mice, trypsin- and serum stimulated larvae, elongated parasites as well as segmented and mature worms. It is unambiguously confirmed that sexually mature worms are obtained as a result of this in vitro process of differentiation. Defined cellular regions are present in all stages of development studied, some of them surrounded by a basal lamina. Glycogen is present in the larvae obtained from the mouse peritoneal cavity and in parasites encapsulated in the mouse host liver. Glycogen distribution in the parasite changes on trypsin and serum stimulation to differentiate. We propose that changes in the distribution of neutral polysaccharides in the parenchyma of the parasite at different stages of development and degradation of polysaccharides in the transition from segmented to adult worm are related to energy needs necessary for the cellular processes leading to the mature specimen. PMID- 19814847 TI - Identification and metabolite profiling of Sitophilus oryzae L. by 1D and 2D NMR spectroscopy. AB - The polyphagous insect Sitophilus oryzae L. (Coleoptera:Curculionidae) has a tremendous adaptability in feeding behaviour, making it a serious invasive pest of stored cereals. The present study identifies the metabolite composition of Sitophilus oryzae (S. oryzae) using Nuclear Magnetic Resonance (NMR) spectroscopy. Assignment of 1D-proton by NMR, 1H-1H COSY, 2D-TOCSY 1H-1H, had been done. Amongst the various biochemically important metabolites isoleucine, valine, leucine, beta-hydroxybutyrate, lysine, glutamate, glutamine, proline, lactate, alanine, di-methylamine, alpha-glucose, beta-glucose, choline, glycerophosphorylcholine and tyrosine are present in S. oryzae. In wheat-fed S. oryzae, the presence of threonine and the absence of lactate is observed. In rice fed S. oryzae, however, the presence of lactate and the absence of threonine were observed. Barley-fed S. oryzae shows presence of both tyrosine and lactate. It is concluded that the pest S. oryzae has adaptability on different stored cereals and grains, depicting the presence of earlier reported metabolites. The present study aims to identify the key metabolic components and associated enzymes in Sitophilus oryzae fed on different cereals. PMID- 19814848 TI - Adult fecundity, host plant preferences, field activity and parasitism in the leaf weevil Phyllobius pyri (L.) (Coleoptera: Curculionidae). AB - Adults of the leaf weevil Phyllobius pyri (L.) (Coleoptera: Curculionidae) feed on a wide variety of broadleaved trees and occasionally cause severe defoliation in newly established farm woodlands. There is little information, however, on the relative susceptibility of different tree species to damage or on the habitat associations of adults and larvae of P. pyri, which might indicate the conditions that predispose trees to attack. Captures of adult P. pyri in emergence and flight traps in the current study indicated population densities in grassland of 0.5-6.4 adults per m2 at emergence but higher densities up to 13.5 per m2 in young pine plantations, where there was a mixture of grassy patches and young, naturally regenerating birch trees. The close proximity of larval food resources (grass roots) and a favoured adult host-plant, which also occurs in young farm woodlands, provided ideal conditions for P. pyri and allowed high population densities to develop. Feeding and performance experiments indicated that cherry, birch, oak and hornbeam were most susceptible to P. pyri, whereas field maple, hawthorn, rowan, lime and especially ash were resistant. Adult female P. pyri emerged in May reproductively immature and fed on tree foliage for 15.9+/-0.9 days before laying their first batch of eggs. Adults lived for 33.3+/-1.5 days, on average, and females laid a mean of 191.9+/-34.5 eggs (maximum=589) during their lifetime. Eggs hatched after 16-20 days. During 2003 and 2004, 11-16% of adult P. pyri were parasitised by Pygostylus falcatus (Hymenoptera: Braconidae) and 19-29% were parasitised by Rondania fasciata (Diptera: Tachinidae). PMID- 19814849 TI - Native range assessment of classical biological control agents: impact of inundative releases as pre-introduction evaluation. AB - Diadromus pulchellus Wesmael (Hymenoptera: Ichneumonidae) is a pupal parasitoid under consideration for introduction into Canada for the control of the invasive leek moth, Acrolepiopsis assectella (Zeller) (Lepidoptera: Acrolepiidae). Since study of the parasitoid outside of quarantine was not permitted in Canada at the time of this project, we assessed its efficacy via field trials in its native range in central Europe. This was done by simulating introductory releases that would eventually take place in Canada when a permit for release is obtained. In 2007 and 2008, experimental leek plots were artificially infested with pest larvae to mimic the higher pest densities common in Canada. Based on a preliminary experiment showing that leek moth pupae were suitable for parasitism up to 5-6 days after pupation, D. pulchellus adults were mass-released into the field plots when the first host cocoons were observed. The laboratory-reared agents reproduced successfully in all trials and radically reduced leek moth survival. Taking into account background parasitism caused by naturally occurring D. pulchellus, the released agents parasitized at least 15.8%, 43.9%, 48.1% and 58.8% of the available hosts in the four release trials. When this significant contribution to leek moth mortality is added to previously published life tables, in which pupal parasitism was absent, the total pupal mortality increases from 60.1% to 76.7%. This study demonstrates how field trials involving environmental manipulation in an agent's native range can yield predictions of the agent's field efficacy once introduced into a novel area. PMID- 19814850 TI - The transmission of nosocomial pathogens in an intensive care unit: a space-time clustering and structural equation modelling approach. AB - We investigated the incidence of cases of nosocomial pathogens and risk factors in an intensive treatment unit ward to determine if the number of cases is dependent on location of patients and the colonization/infection history of the ward. A clustering approach method was developed to investigate the patterns of spread of cases through time for five microorganisms [methicillin-resistant Staphylococcus aureus (MRSA), Acinetobacter spp., Klebsiella spp., Candida spp., and Pseudomonas aeruginosa] using hospital microbiological monitoring data and ward records of patient-bed use. Cases of colonization/infection by MRSA, Candida and Pseudomonas were clustered in beds and through time while cases of Klebsiella and Acinetobacter were not. We used structural equation modelling to analyse interacting risk factors and the potential pathways of transmission in the ward. Prior nurse contact with colonized/infected patients, mediated by the number of patient-bed movements, were important predictors for all cases, except for those of Pseudomonas. General health and invasive surgery were significant predictors of cases of Candida and Klebsiella. We suggest that isolation and bed movement as a strategy to manage MRSA infections is likely to impact upon the incidence of cases of other opportunist pathogens. PMID- 19814851 TI - Rabies epidemiology and control in Turkey: past and present. AB - SUMMARYTurkey is the only country in Europe where urban dog-mediated rabies persists. Control measures in recent decades have reduced the burden of rabies to relatively low levels but foci of disease still persist, particularly in urban areas. Occasional human cases result from this persistence although the source of these appears to be both dog and wildlife reservoirs. This review considers the current state of rabies in Turkey including current control measures, the varying epidemiology of the disease throughout this country and the prospects for rabies elimination. PMID- 19814852 TI - A prospective study of diurnal cortisol and cognitive function in community dwelling elderly people. AB - BACKGROUND: Elevated cortisol levels due to hypothalamic-pituitary-adrenal (HPA) axis stress response have been associated with cognitive impairment. However, the causal relationship between stress and subsequent cognitive impairment remains unclear, notably because of the small number of gender-stratified prospective studies. METHOD: Salivary cortisol secretion was evaluated in 197 non-depressed community-dwelling elderly people at three time points on the day of hospital attendance for a clinical examination and again on the following day at home, in a distinct environmental context. Cognitive performance was evaluated at baseline and at 2- and 4-year follow-up. RESULTS: Cross-sectional logistic analyses adjusted for age and education indicated that men with high morning cortisol at the hospital had higher risk of low cognitive performance in verbal fluency [odds ratio (OR) 3.0, p=0.05] and visuospatial performance (OR 5.1, p=0.03). Impairment in verbal fluency was observed in women with moderate high morning cortisol (OR 3.6, p=0.05) or moderate slow diurnal rhythm (OR 3.7, p=0.04). In longitudinal analyses, slow diurnal rhythm (flatter slope) was associated with decline over 4 years in visuospatial performance (OR 7.7, p=0.03) and visual memory (OR 4.1, p=0.03) in men, and in verbal fluency (OR 6.0, p=0.01) in women. High morning cortisol was associated with decline in visual memory in women (OR 5.1, p=0.06). CONCLUSIONS: HPA axis dysregulation seems to be associated with low cognitive performance in the elderly. Slower cortisol elimination rates could predict cognitive decline affecting principally non-verbal functioning in men and verbal functioning in women. The effects are independent of environmental context, apolipoprotein E (ApoE) genotype or psychopathology. Interventions blocking this pathway may provide new therapeutic options to prevent cognitive decline. PMID- 19814853 TI - Food insecurity - not just about rural communities in Africa and Asia. PMID- 19814854 TI - Food insecurity: the skeleton in the national closet. PMID- 19814855 TI - Do food regulatory systems protect public health? PMID- 19814856 TI - Marketing of unhealthy food to young children: Brazilian Goliath skulking. PMID- 19814857 TI - Importance of pharmaceutical composition and evidence from clinical trials and pharmacological studies in determining effectiveness of chondroitin sulphate and other glycosaminoglycans: a critique. AB - OBJECTIVES: Chondroitin sulphate (CS) has attracted much interest over the past two decades or so as a biological agent for use in the relief of pain and joint symptoms in osteoarthritis. Earlier clinical investigations produced variable, if encouraging results. This variability was partly due to limitations on the study designs and the lack of availability of standardized CS. Recently, high quality and fully standardized CS (Condrosulf) has become available and its effects have been studied in large-scale osteoarthritis trials, which are discussed here. KEY FINDINGS: There is now evidence for symptom- and structure-modifying (radiologically-observed) effects. These studies show that CS (a) has slow onset of response and that relief of pain may not be like that of the direct analgesic actions of non-steroidal anti-inflammatory drugs (NSAIDs), (b) there are indications of reduced need for intake of analgesics (e.g. NSAIDs) in patients taking CS, and (c) quality of life and cost-benefits may be associated with use of CS. Safety evaluations show that the incidence of adverse reactions is low. Pharmacokinetic studies indicate that although oral absorption is relatively fast CS has moderate oral bioavailability (15-24%) and that depolymerised and degraded CS that is evident after absorption, together with CS itself, may take some time to accumulate in target joints. The pharmacodynamic actions of CS indicate that it has anti-inflammatory effects that include multiple actions involving reduction of catabolic reactions and enhanced anabolic (proteoglycan) synthetic reactions in cartilage and may block osteoclast activation in bone. Further studies are required to (a) establish the effects of depolymerised and degraded CS on degradation of cartilage and bone in vitro, and (b) MRI and other investigations of the effects in osteoarthritis of long-term CS treatment. SUMMARY: The findings from this review show there may be potential value of CS in reducing the dependence on intake of NSAIDs and analgesics in patients with osteoarthritis, while at the same time having favourable safety. PMID- 19814858 TI - Quality of different chondroitin sulfate preparations in relation to their therapeutic activity. AB - OBJECTIVES: Chondroitin sulfate is currently recommended by the European League Against Rheumatism (EULAR) as a SYSADOA (symptomatic slow acting drug for osteoarthritis) in Europe in the treatment of knee and hand osteoarthritis based on research evidence and meta-analysis of numerous clinical studies. Furthermore, recent clinical trials demonstrated its possible structure-modifying effects. Chondroitin sulfate, alone or in combination with glucosamine or other ingredients, is also utilized as a nutraceutical in dietary supplements in Europe and the USA. However, it is derived from animal sources by extraction and purification processes. As a consequence, source material, manufacturing processes, the presence of contaminants and many other factors contribute to the overall biological and pharmacological actions of these agents. We aim to review the quality control of chondroitin sulfate in pharmaceutical-grade preparations and nutraceuticals. KEY FINDINGS: Pharmaceutical-grade formulations of chondroitin sulfate are of high and standardized quality, purity and properties, due to the stricter regulations to which this drug is subjected by local national health institutes as regards production and characteristics. On the contrary, as several published studies available in literature indicate, the chondroitin sulfate quality of several nutraceuticals is poor. Additionally, there are no definite regulations governing the origin of the ingredients in these nutraceuticals and the origin of the ingredients in natural products is the most important factor ensuring quality, and thus safety and efficacy, in particular for chondroitin sulfate, due to its extraction from different sources. CONCLUSIONS: Due to the poor chondroitin sulfate quality of some nutraceuticals, we conclude that stricter regulations regarding their quality control should be introduced to guarantee the manufacture of high quality products for nutraceutical utilization and to protect customers from low-quality, ineffective and potentially dangerous products. There is a need for specific and accurate analytical procedures, which should be enforced to confirm purity and label claims both for raw materials and finished chondroitin sulfate products, and also to govern the origin of ingredients. Until these stricter regulations are in place, then it is strongly recommended that pharmaceutical-grade chondroitin sulfate is used rather than food supplements. PMID- 19814859 TI - Boswellia resin: from religious ceremonies to medical uses; a review of in-vitro, in-vivo and clinical trials. AB - OBJECTIVES: Despite its historical-religious, cultural and medical importance, Boswellia has not been thoroughly studied, and gaps still exist between our knowledge of the traditional uses of the resin and the scientific data available. Here we review the pharmacology of Boswellia resin and of the small molecules identified as the active ingredients of the resin. KEY FINDINGS: The resin of Boswellia species ('frankincense', 'olibanum') has been used as incense in religious and cultural ceremonies since the beginning of written history. Its medicinal properties are also widely recognized, mainly in the treatment of inflammatory conditions, as well as in some cancerous diseases, wound healing and for its antimicrobial activity. Until recently, work on Boswellia focused on the immunomodulatory properties of the resin and boswellic acids were considered to be the main, if not the only, active ingredients of the resin. Hence, this family of triterpenoids was investigated by numerous groups, both in vitro and in vivo. These compounds were shown to exert significant anti-inflammatory and pro apoptotic activity in many assays: in vitro, in vivo and in clinical trials. We recently found incensole acetate and its derivatives, which are major components of Boswellia resin, to be nuclear factor-kappaB inhibitors, thus suggesting that they are, at least in part, responsible for its anti-inflammatory effects. Incensole acetate also exerts a robust neuroprotective effect after brain trauma in mice. Furthermore, it causes behavioural as well as anti-depressive and anxiolytic effects in mice. It is also a potent agonist of the transient receptor potential (TRP)V3 channel. It thus seems that incensole acetate and its derivatives play a significant role in the effects that Boswellia resin exerts on biological systems. CONCLUSIONS: Altogether, studies on Boswellia resin have provided an arsenal of bio-active small molecules with a considerable therapeutic potential that is far from being utilized. PMID- 19814860 TI - Arsthinol nanosuspensions: pharmacokinetics and anti-leukaemic activity on NB4 promyelocytic leukaemia cells. AB - OBJECTIVES: The organoarsenical arsthinol was used in the 1950s in the treatment of amoebiasis and yaws and was considered as 'highly tolerated'. The aim of this work was to study its anti-leukaemic activity and to develop nanosuspensions of the drug, thereby limiting brain concentrations and the risk of encephalopathy. METHODS: Arsthinol nanosuspensions were produced by high-pressure homogenization. The anti-leukaemic activity was assessed on NB4 acute promyelocytic leukaemia cells (vs solutions of arsthinol, As(2)O(3) and melarsoprol). In addition, a pharmacokinetics study was performed to compare the nanosuspensions and the solution of arsthinol. KEY FINDINGS: Arsthinol induced growth inhibition of NB4 cells at lower concentration (IC50 (concentration inhibiting growth by 50%) = 0.78 +/- 0.08 micromol/l after 24 h) than As(2)O(3) (IC50 = 1.60 +/- 0.23 micromol/l after 24 h) or melarsoprol (IC50 = 1.44 +/- 0.08 micromol/l after 24 h). When formulated as nanosuspension, arsthinol remained cytotoxic (IC50 = 1.33 +/- 0.30 micromol/l after 24 h). This formulation also reduced the drug's access to the brain (C(max) = 0.03 micromol/g) whereas bone marrow concentrations remained very high (C(max) = 2 micromol/g). CONCLUSIONS: Nanosuspensions of arsthinol could be proposed for further studies in the treatment of acute promyelocytic leukaemia. PMID- 19814861 TI - In-vitro interaction of L-dopa with bacterial adhesins of Helicobacter pylori: an explanation for clinicial differences in bioavailability? AB - OBJECTIVES: Recent investigations on the pharmacokinetics of levodopa (L-dopa) indicated that the presence of Helicobacter pylori in patients with Parkinson's disease, orally treated with L-dopa, influences the absorption of this compound, which consequently leads to decreased plasma levels. Therefore this work aims to study a potential in-vitro interaction of L-dopa with H. pylori and its surface adhesins. METHODS: Solutions containing L-dopa of different concentrations were incubated with H. pylori at different bacterial densities and time intervals. Free L-dopa was quantified from the incubation supernatants by HPLC. A flow cytometric assay with fluorescence labelled H. pylori was used to investigate the influence of L-dopa on the bacterial adhesion of H. pylori: FITC-labelled bacteria were pre-incubated with L-dopa, followed by incubation with gastric epithelial cells (AGS cells) and FACS quantification of adhering bacteria. KEY FINDINGS: Evaluation of time- and concentration-dependent incubation experiments indicated a significant decrease in L-dopa concentrations when coming into contact with H. pylori. The reduction in L-dopa concentrations was determined as 47 to 12%, referred to the initial starting concentration, with time-dependency and dependency of the H. pylori density. FITC-labelled H. pylori, pre-incubated with differing L-dopa concentrations, were shown to have a significantly reduced bacterial adhesion to AGS cells, with a maximum reduction of 22 +/- 9%. These results demonstrate a direct interaction of L-dopa with the outer membrane proteins of H. pylori responsible for the adhesion to gastric epithelial cells. By this interaction the unbound L-dopa concentration in bacterial suspension was strongly reduced. CONCLUSIONS: This study suggests a potential in-vitro interaction of L-dopa with H. pylori adhesins, confirming the clinical changes found in pharmacokinetics of L-dopa therapy by H. pylori-positive patients with Parkinson's disease. PMID- 19814862 TI - In-vitro and in-vivo metabolic studies of the candidate chemopreventative pentamethylchromanol using liquid chromatography/tandem mass spectrometry. AB - OBJECTIVES: This study focuses on the in-vitro metabolic profiles of pentamethylchromanol in human, rat, dog and non-human primates, and characterizes the associated metabolic kinetics and specific human isozymes responsible for metabolism. Additional investigations compare in-vitro data with in-vivo metabolic data from rats and dogs. METHODS: In-vitro metabolites were generated from commercially available microsomes, S9 fractions and cytochrome P450 isozymes. Reaction mixtures were analysed using liquid chromatography/tandem mass spectrometry for metabolite identification, stability, phenotyping and kinetic profiles. Plasma samples were collected from 28-day toxicology studies in rats and dogs, and analysed using the same methodology as for the identification of in vitro metabolites. KEY FINDINGS: Samples from in-vitro experiments produced a total of eight identified metabolites while five were observed in the in-vivo samples. Kinetic analysis of metabolites in human microsomes generated Michaelis constants (K(M)) ranging from 10.9 to 104.9 mum. Pentamethylchromanol metabolic stability varied by species and multiple isozymes were identified for the observed biotransformation pathways. Pentamethylchromanol is susceptible to multiple metabolic pathways and differential metabolic stability, which is species dependent. CONCLUSIONS: In-vitro metabolism was not a strong predictor of in-vivo metabolism for the samples assays but showed glucuronidation and sulfation as common biotransformation pathways. PMID- 19814863 TI - Effect of intestinal ischaemia/reperfusion on P-glycoprotein-mediated ileal excretion of rhodamine 123 in the rat. AB - OBJECTIVES: We have shown that ischaemia/reperfusion in the small intestine at an early phase, such as 1 h after reperfusion, induced not only functional changes in the membrane, such as P-glycoprotein (P-gp) dysfunction, but also decreased expression of P-gp protein and mdr1a mRNA. In the present study we examined whether intestinal ischaemia/reperfusion modifies the P-gp-mediated ileal excretion transport system in rats beyond 1 h after reperfusion. METHODS: To evaluate the contribution of P-gp-mediated transport to the ileal excretion of rhodamine 123, we used Western blotting to measure the expression of P-gp protein levels isolated from the ileum at different reperfusion times after 60 min of ischaemia. We also measured the expression of inducible nitric oxide synthase (iNOS) mRNA using real-time RT-PCR. KEY FINDINGS: Ileal excretion of rhodamine 123 decreased at 3 h after reperfusion and had recovered at 24 h. Changes in villi structure at 3 h and its recovery at 24 h were also observed. Verapamil, a competitive inhibitor of P-gp, significantly inhibited ileal clearance of rhodamine 123 to the lumen at 24 h after reperfusion, suggesting that P-gp was working at this time. These results suggest that intestinal ischaemia/reperfusion induced decrease in P-gp-mediated ileal excretion of rhodamine 123 was probably due to impaired P-gp-mediated transport. Levels of P-gp protein and iNOS mRNA in the ileum decreased 3 h after ischaemia/reperfusion and returned to control levels after 24 h. CONCLUSIONS: These findings suggest that intestinal ischaemia/reperfusion markedly decreases P-gp-mediated ileal excretion of rhodamine 123, probably by decreasing the expression of P-gp protein, which is likely to be due to increased lipid peroxidation via iNOS. PMID- 19814864 TI - Faster clearance of mirodenafil in rats with acute renal failure induced by uranyl nitrate: contribution of increased protein expression of hepatic CYP3A1 and intestinal CYP1A1 and 3A1/2. AB - OBJECTIVES: It has been reported that mirodenafil is primarily metabolized via hepatic cytochrome P450 (CYP) 1A1/2, 2B1/2, 2D1 and 3A1/2 in rats. It has also been reported that the protein expression of hepatic CYP3A1 and intestinal CYP1A1 and 3A1/2 increases and that of hepatic CYP2D1 decreases in rats with acute renal failure induced by uranyl nitrate (U-ARF rats). Thus, the pharmacokinetics of mirodenafil were studied in control and U-ARF rats. METHODS: The pharmacokinetic parameters of mirodenafil and SK3541 (a metabolite of mirodenafil) were compared after the intravenous and oral administration of mirodenafil at a dose of 20 mg/kg to U-ARF and control rats. KEY FINDINGS: After interavenous administration of mirodenafil to U-ARF rats, the total area under the concentration-time curve (AUC) of mirodenafil was significantly smaller (36.5% decrease) than controls, possibly due to the significantly faster non-renal clearance (66.1% increase; because of increase in the protein expression of hepatic CYP3A1) than controls. After the oral administration of mirodenafil to U-ARF rats, the AUC of mirodenafil was also significantly smaller (47.8% decrease) due to the increase in the protein expression of hepatic CYP3A1 and intestinal CYP1A1 and 3A1/2 compared with controls. CONCLUSIONS: After both intravenous and oral administration of mirodenafil to U-ARF rats, the AUC(SK3541)/AUC(mirodenafil) ratios were comparable with that in controls and this could be due to further metabolism of SK3541 in rats. PMID- 19814865 TI - Therapeutic index of methotrexate depends on circadian cycling of tumour necrosis factor-alpha in collagen-induced arthritic rats and mice. AB - OBJECTIVES: Rheumatoid arthritis is an autoimmune disorder of unknown aetiology. Morning stiffness, a characteristic feature of rheumatoid arthritis, shows a 24-h rhythm. Noticing this rhythm, we hypothesized the presence of a similar rhythm for a rheumatoid arthritis indicator, in addition to dosing-time dependency of the anti-rheumatic effect of methotrexate in arthritis induced by collagen in rats and mice, which reflect the symptomatology of rheumatoid arthritis patients. METHODS: To measure tumour necrosis factor (TNF)-alpha concentration, blood was taken at different times (2, 6, 10, 14, 18 or 22 h after the light was turned on (HALO)) in collagen-induced arthritic mice. Methotrexate was administered at two different dosing times based on these findings to estimate arthritis. KEY FINDINGS: The arthritis score was significantly lower in the 22 HALO-treated group than in the control and 10 HALO-treated groups in collagen-induced arthritic rats and mice. Plasma TNF-alpha concentrations showed obvious 24-h rhythms, with higher levels at light phase and lower levels at dark phase after rheumatoid arthritis crisis. Arthritis was relieved after administration of methotrexate during the dark phase in synchronization with the 24-h rhythm. CONCLUSIONS: Our findings suggest that choosing an optimal dosing time associated with the 24-h cycling of TNF-alpha could lead to effective treatment of rheumatoid arthritis by methotrexate. PMID- 19814866 TI - Comparative study between the effect of the peroxisome proliferator activated receptor-alpha ligands fenofibrate and n-3 polyunsaturated fatty acids on activation of 5'-AMP-activated protein kinase-alpha1 in high-fat fed rats. AB - OBJECTIVES: Obesity is a risk factor for type 2 diabetes mellitus. It results from an energy imbalance in which energy intake exceeds energy expenditure. The cellular fuel gauge 5'-AMP-activated protein kinase (AMPK) is a heterotrimeric protein consisting of one catalytic subunit (alpha) and two non-catalytic subunits (beta and gamma), and approximately equal levels of alpha1 and alpha2 complexes are present in the liver. AMPK regulates metabolic pathways in response to metabolic stress and in particular ATP depletion to switch on energy-producing catabolic pathways such as beta-oxidation of fatty acids and switch off energy depleting processes such as synthesis of fatty acid and cholesterol. A high-fat diet alters AMPK-alpha1 gene expression in the liver and skeletal muscle of rats and results in body weight gain and hyperglycaemia. The aim of this study was to investigate and compare the potential effects of peroxisome proliferator activated receptor (PPAR)-alpha agonists fenofibrate and n-3 polyunsaturated fatty acids (PUFAs) in modulation of AMPK-alpha1 activity in liver and skeletal muscle of high-fat diet fed rats. METHODS: Reverse transcription-polymerase chain reaction was used for determination of AMPK-alpha1 in liver and soleus muscle and both PPAR-alpha and CPT-1 in hepatic tissues. Serum, total cholesterol, triacylglycerol, fatty acid and fasting blood glucose were determined colorimetrically. KEY FINDINGS: Both PPAR-alpha agonists, fenofibrate and n-3 PUFA, increased the mRNA expression of AMPK-alpha1 activity in liver and skeletal muscle of obese diabetic rats. Fenofibrate was superior in its activation of hepatic mRNA expression of AMPK-alpha 1 to exert more lipolytic effect and body weight reduction, as estimated through the decrease of triacylglycerol output and serum levels of fatty acid on the one hand and the increase in CPT-1 mRNA expression, the key enzyme in beta-oxidation of fatty acid, on the other hand. n 3 PUFA activated AMPK-alpha1 mRNA expression in skeletal muscle much more than fenofibrate to reveal more hypoglycaemic effect. CONCLUSIONS: The PPAR-alpha agonists fenofibrate and n-3 PUFA could efficiently activate AMPK-alpha1 mRNA expression in liver and skeletal muscle to exert body weight reduction and hypoglycaemic effect, respectively. PMID- 19814867 TI - Acteoside inhibits alpha-MSH-induced melanin production in B16 melanoma cells by inactivation of adenyl cyclase. AB - OBJECTIVES: The aim of the study was to determine the mechanism of the whitening effect of acteoside. METHODS: We used tyrosinase activity and melanin production stimulated in B16 melanoma cells by alpha-melanocyte stimulating hormone (alpha MSH) or forskolin to measure the whitening effect of acteoside. KEY FINDINGS: Acteoside did not directly inhibit mushroom tyrosinase activity, but dose dependently inhibited tyrosinase activity and melanin production in B16 melanoma cells stimulated by 1 micromol/l alpha-MSH. Acteoside also reduced cyclic AMP levels in cells stimulated by 1 micromol/l alpha-MSH, suggesting direct inhibition of adenyl cyclase. Acteoside also inhibited production of both melanin and cyclic AMP in cells stimulated by 1 micromol/l forskolin, an adenyl cyclase activator. Acteoside showed antioxidant activity in a cell-free DPPH (1-diphenyl 2-picrylhydroazyl) assay and inhibited generation of intracellular reactive oxygen species. CONCLUSIONS: These results suggest that the whitening activity of acteoside results from inhibition of adenyl cyclase and alpha-MSH signalling. PMID- 19814868 TI - Enhanced anticancer activity of glutamate prodrugs of all-trans retinoic acid. AB - OBJECTIVES: All-trans retinoic acid (ATRA), an active metabolite of vitamin A, is widely used in the treatment of acute promyelocytic leukaemia and myelodysplastic syndrome. However, its high lipophilicity is thought to be responsible for the slow dissolution and low bioavailability following oral administration. In order to obtain compounds with better solubility characteristics to improve the transportation and bioavailability of ATRA, derivatives of ATRA containing glutamic acid or its sodium salt were synthesised. METHODS: The ATRA derivatives synthesised - all-trans retinoyl glutamate (RAE) and all-trans retinoyl sodium glutamate (RAENa(2)) - were characterised in terms of melting point, optical rotation, mass spectrometry, NMR and partition coefficient. A liposomal preparation formed from RAE was characterised by particle size and zeta potential. The anti-tumour activity of RAE and RAENa(2) was compared with that of ATRA in mice bearing S(180) tumours and their effects on the cell cycle were determined in human pro-myelocytic leukaemia HL-60 cells. KEY FINDINGS: RAE and RAENa(2) were more active than ATRA against tumour growth. Flow cytometry indicated that RAE and RAENa(2) induced HL-60 cell cycle arrest, similar to ATRA. DNA fragmentation studies suggested that apoptosis may be one of the mechanisms responsible for the anti-tumour activities. CONCLUSIONS: The two derivatives of ATRA, RAE and RAENa(2), exhibited improved aqueous solubility and were more effective in mice bearing S(180) tumours. PMID- 19814869 TI - Intestinal inflammation and seizure susceptibility: understanding the role of tumour necrosis factor-alpha in a rat model. AB - OBJECTIVES: The aim of the study was to evaluate the correlation between colitis and susceptibility to seizures. METHODS: Colitis was induced in Wistar rats by a single intracolonic administration of trinitrobenzene sulfonic acid (TNBS; 20 mg in 35% ethanol). The control group were given intracolonic vehicle. One group of rats with colitis were treated with thalidomide (150 mg/kg p.o.) daily for 14 days. The other colitis group received vehicle only. On day 15, seizure susceptibility was tested by administration of pentylenetetrazole (40 mg/kg i.p.). Colonic tissue was collected for estimation of morphological score, and malondialdehyde, superoxide dismutase, catalase and glutathione peroxidase. Tumour necrosis factor (TNF)-alpha levels were measured in serum and brain samples. KEY FINDINGS: The colitis group showed a significant increase in seizure score and reduction in onset time compared with the control group. Thalidomide was protective against seizures, resulting in decreased seizure score and significantly delaying the onset of seizures. Thalidomide also provided significant protection against TNBS-induced colonic damage in terms of morphological and histological score and levels of lipid peroxidation, superoxide dismutase, catalase and glutathione peroxidase in colonic tissue. The level of TNF-alpha in serum was also reduced significantly whereas brain TNF-alpha level was reduced but not significantly. CONCLUSIONS: TNBS-induced colitis increased seizure susceptibility to a subconvulsive dose of pentylenetetrazole; the immunomodulator thalidomide was protective. PMID- 19814870 TI - Cardioprotective actions of two bioflavonoids, quercetin and rutin, in experimental myocardial infarction in both normal and streptozotocin-induced type I diabetic rats. AB - OBJECTIVES: Revascularization therapy is the mainstay of treatment in the management of myocardial infarction in normal and diabetic patients. We attempted to evaluate the cardioprotective actions of quercetin and rutin in ischaemia reperfusion-induced myocardial infarction in both normal and diabetic rats. METHODS: Myocardial infarct size was measured using the staining agent 2,3,5 triphenyltetrazoliumchloride. Serum and tissue malondialdehyde levels and superoxide dismutase and catalase in heart tissue were estimated spectrophotometrically. A lead II electrocardiogram was monitored at various intervals throughout the experiment. KEY FINDINGS: Results demonstrated the larger infarct size, enhanced lipid peroxidation, partial depletion of antioxidant enzymes and drastic drop in heart rate in diabetic hearts subjected to in-vivo ischaemia-reperfusion in comparison to normal rats subjected to ischaemia-reperfusion. Furthermore, quercetin and rutin significantly limit the infarct size in both normal and diabetic animals in a similar fashion. However, rutin offered complete cardioprotection at a dose of 10 mg/kg in terms of limiting infarct size. Both flavonoids could partially but significantly attenuate the lipid peroxidation. In addition, treatment has shown moderate improvement in heart rate in both normal and diabetic rats. CONCLUSIONS: Our data suggest the possible cardioprotective effects of quercetin and rutin in ischaemia reperfusion injury in both normal and diabetic rats, and that protection might be in part due to the attenuation of oxidative stress and moderate increment in antioxidant reserves. PMID- 19814871 TI - Hepatoprotective effects of salidroside on fulminant hepatic failure induced by D galactosamine and lipopolysaccharide in mice. AB - OBJECTIVES: The aim was to investigate the protective effect of salidroside isolated from Rhodiola sachalinensis A. Bor. (Crassulaceae) on D galactosamine/lipopolysaccharide-induced fulminant hepatic failure. METHODS: Hepatotoxicity was induced by an intraperitoneal injection of D-galactosamine (700 mg/kg) and lipopolysaccharide (10 mug/kg); salidroside (20, 50 and 100 mg/kg) was administered intraperitoneally 1 h before induction of hepatoxicity. Liver injury was assessed biochemically and histologically. KEY FINDINGS: Salidroside attenuated the induced acute increase in serum aspartate aminotransferase and alanine aminotransferase activities, and levels of tumour necrosis factor-alpha levels and serum nitric oxide. It restored depleted hepatic glutathione, superoxide dismutase, catalase and glutathione peroxidase activities, decreased malondialdehyde levels and considerably reduced histopathological changes. Histopathological, immunohistochemical and Western blot analyses also demonstrated that salidroside could reduce the appearance of necrotic regions and expression of caspase-3 and hypoxia-inducible factor-1alpha in liver tissue. CONCLUSIONS: Salidroside protected liver tissue from the oxidative stress elicited by D-galactosamine and lipopolysaccharide. The hepatoprotective mechanism of salidroside appear to be related to antioxidant activity and inhibition of hypoxia-inducible factor-1alpha. PMID- 19814872 TI - Potential beneficial effect of naringenin on lipid peroxidation and antioxidant status in rats with ethanol-induced hepatotoxicity. AB - OBJECTIVES: The aim was to study the effect of naringenin, a biologically active compound, on tissue antioxidant status and lipid peroxidation in ethanol-induced hepatotoxicity in rats. METHODS: Rats were divided into four groups: Groups 1 and 2 received isocaloric glucose and 0.5% carboxymethyl cellulose; groups 3 and 4 received 20% ethanol equivalent to 6 g/kg daily for 60 days. In addition, groups 2 and 4 were given naringenin (50 mg/kg) daily for the last 30 days of the experiment. KEY FINDINGS: The results showed significantly elevated levels of serum aspartate and alanine transaminases, gamma-glutamyl transpeptidase, tissue thiobarbituric acid reactive substances, conjugated dienes, lipid hydroperoxides and protein carbonyl content, and significantly lowered activities/levels of antioxidants such as superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione-S-transferase, reduced glutathione and vitamins C and E in ethanol-treated rats compared with control rats. Administration of naringenin to rats with ethanol-induced liver injury significantly decreased the levels of serum aspartate and alanine transaminases, gamma-glutamyl transpeptidase, tissue thiobarbituric acid reactive substances, conjugated dienes, lipid hydroperoxides and protein carbonyl content and significantly elevated the activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glutathione-S-transferase, and the levels of reduced glutathione and vitamins C and E in the tissues compared with unsupplemented ethanol-treated rats. Histological changes observed in the liver correlated with the biochemical findings. CONCLUSIONS: Taken together these findings suggest that naringenin has a therapeutic potential in the abatement of ethanol-induced hepatotoxicity. PMID- 19814874 TI - Penetration of oseltamivir and its active metabolite into the brain after lipopolysaccharide-induced inflammation in mice. AB - OBJECTIVES: Oseltamivir phosphate is used for the treatment of influenza virus infections. Recently, oral intake has been associated with abnormal behaviour. The present study examined the brain penetration of oseltamivir phosphate and oseltamivir carboxylate, its active metabolite, during inflammation. METHODS: Male C57BL/6 mice were given three i.p. injections of lipopolysaccharide (LPS) or saline. We studied the concentration of Evans blue (a marker of blood-brain barrier function) and oseltamivir phosphate and its active metabolite in the brain and plasma. KEY FINDINGS: The brain-to-plasma ratio of Evans blue compared with saline-treated control mice increased significantly with LPS dose. LPS induced inflammation increased the permeation of drugs through the blood-brain barrier. The concentration of oseltamivir phosphate in both brain and plasma was 2-fold higher in mice treated with LPS than in control mice. Although the plasma concentration of the active carboxylate was not significantly altered by inflammation, the brain concentration was increased 2.7-fold in mice treated with LPS compared with control mice. CONCLUSIONS: Administration of oseltamivir phosphate in the presence of inflammation increased the brain concentration of both parent drug and active metabolite, which may explain the central nervous system side-effects observed with this agent. PMID- 19814873 TI - Evaluation of anti-angiogenic, anti-inflammatory and antinociceptive activity of coenzyme Q(10) in experimental animals. AB - OBJECTIVES: This work aimed to assess some pharmacological activities of coenzyme Q(10) (CoQ(10)) in animal experimental models. METHODS: The chick chorioallantoic membrane assay was used to evaluate anti-angiogenic activity of CoQ(10). Anti inflammatory activity of CoQ(10) was confirmed using two animal models of inflammation. These were the vascular permeability and air pouch models, models of acute and sub-acute inflammation, respectively. Antinociceptive activity was assessed by the acetic acid-induced abdominal constriction response. KEY FINDINGS: CoQ(10) dose-dependently displayed inhibition of chick chorioallantoic membrane angiogenesis. In the acetic acid-induced vascular permeability model in mice, CoQ(10) at 50, 100 and 200 mg/kg reduced vascular permeability from 0.74 +/ 0.01 (A(590)) to 0.67 +/- 0.01 (P < 0.01), 0.46 +/- 0.02 (P < 0.01) and 0.30 +/- 0.01 (P < 0.01), respectively. In the carrageenan-induced inflammation in the air pouch, CoQ(10) was able to diminish exudate volume, the number of polymorphonulcear leucocytes and nitrite content in the air pouches. CoQ(10) at 25, 50 and 100 mg/kg significantly reduced acetic acid-induced abdominal constriction in mice from 27.0 +/- 2.00 (number of abdominal constrictions) to 17.7 +/- 0.33 (P < 0.01), 9.3 +/- 0.67 (P < 0.01) and 1.3 +/- 0.33 (P < 0.01), respectively, suggesting a strong antinociceptive activity. CONCLUSIONS: CoQ(10) possessed considerable anti-angiogenic, anti-inflammatory and antinociceptive activity, possibly via down-regulating the level of nitric oxide, which partly supported its use as a dietary supplement and in combination therapy. PMID- 19814875 TI - [Associations between geohelminths and socioenvironmental conditions among different human populations in Argentina]. AB - OBJECTIVES: To analyze the relationship between the geohelminth species found in urban, suburban, and rural areas of the Buenos Aires and Misiones provinces of Argentina, and the socioenvironmental conditions that promote infection by these parasites. METHODS: This was a cross-sectional study that analyzed 700 human fecal samples taken from 319 families residing in an urban population (UP) and two suburban ones (SUP1, SUP2) in the Province of Buenos Aires, and a rural one in Misiones (RP). Thirty-five samples of dog feces and 205 earth samples were taken, plus surveys were performed of the socioenvironmental characteristics of the study areas. The Ritchie, Carles-Barthelemy, Fulleborn, and Kato-Katz parasite analysis techniques were used. RESULTS: The highest incidence of parasites was in RP (78.4%), followed by the suburban areas SUP1 (35.0%) and SUP2 (25.8%), and lastly, the urban area (5.7%). Ancylostomatidae (71.1%) and Strongyloides stercoralis (22.2%) were found only in RP, while Ascaris lumbricoides, Hymenolepis nana, and Trichuris trichiura were most frequent in SUP1. The samples of dog feces from the Misiones province had the highest incidence of parasites (100%) compared to those from Buenos Aires, but the urban dirt from this city proved to be more contaminated. Poor hygiene habits in food preparation and toileting, overcrowding, walking barefoot, and dwellings with dirt floors were significantly associated with a higher frequency of intestinal geohelminths and pseudogeohelminths (P < 0.05). CONCLUSIONS: The great number of cases of multiple infestations seen among suburban as well as rural populations calls for special attention and underscores the need for more extensive epidemiological studies that take on the different aspects of this complex issue with the goal of implementing more efficient health initiatives and programs. PMID- 19814876 TI - Estimating diabetes and diabetes-free life expectancy in Mexico and seven major cities in Latin America and the Caribbean. AB - OBJECTIVES: To estimate diabetes and diabetes-free life expectancy in seven major cities in Latin America and the Caribbean, plus Mexico as a whole. METHODS: Data from the Survey on Health, Well-being, and Aging in Latin America and the Caribbean (n = 10 602) and the Mexican Health and Aging Study (n = 6 953) on individuals 60 or more years of age were used in this study. Estimates of diabetes and diabetes-free life expectancy were obtained by applying the Sullivan method. RESULTS: Diabetes life expectancy for men 60 years of age was highest in Mexico City (4.5 years) and Bridgetown (3.4 years), and lowest in Havana (1.3 years). Diabetes-free life expectancy for men 60 years of age was highest in Santiago (17.6 years) and lowest in Bridgetown (14.2 years) and Sao Paulo (14.3 years). For women, diabetes life expectancy was highest in Bridgetown (5.4 years), followed by Mexico City and Havana; but these three cities also had the lowest diabetes-free life expectancy. Women 60 years of age in Buenos Aires had the lowest diabetes life expectancy (2.5 years), and in Santiago, the highest, with a diabetes-free life expectancy of 20.7 years. CONCLUSIONS: Older individuals in Latin America and the Caribbean can expect to live a large proportion of their remaining lives with diabetes. There were also important differences across settings; in particular, the pronounced diabetes burden in Barbados and Mexico and among women. Given the fast growth of the elderly population in these societies, it is crucial to promote healthy eating and exercise as a way of reducing the burden of diabetes. PMID- 19814877 TI - Prevalence and social distribution of risk factors for chronic noncommunicable diseases in Brazil. AB - OBJECTIVE: To assess risk factors for chronic noncommunicable disease (CND) and to identity social inequalities in their distribution among the adult Brazilian population. METHODS: Study of CND risk factors (including tobacco use, overweight and obesity, low fruit and vegetable intake (LFVI), insufficient leisure-time physical activity (LTPA), sedentary lifestyle, and alcohol abuse, among other risks) in a probabilistic sample of 54 369 individuals from Brazil's 26 state capitals and Federal District, in 2006, using the Surveillance System of Risk and Protective Factors for Chronic Non-Communicable Diseases through Telephone Interviews (VIGITEL), a computer-assisted telephone interviewing (CATI) survey system, and calculated age-adjusted prevalence ratios for trends in education levels using Poisson regression with linear models. RESULTS: Men reported higher tobacco use, overweight, LFVI, sedentary lifestyle, and alcohol abuse versus women, but lower insufficient LTPA. In men, education was associated with increased overweight and sedentary lifestyle, but decreased tobacco use, LFVI, and insufficient LTPA. Among women, education was associated with decreased tobacco use, overweight, obesity, LFVI, and insufficient LTPA, but increased sedentary lifestyle. CONCLUSION: In Brazil, prevalence of CND risk factors (except insufficient LTPA) is higher in men. For both sexes, the CND risk factor prevalence ratio is influenced by level of education. PMID- 19814878 TI - [Death from violent causes and economic cycles in Bogota, Colombia: a time-series study, 1997-2006]. AB - OBJECTIVES: To determine seasonal trends in the number of deaths from violence in Bogota, Colombia from 1997-2006 and to analyze any possible associations with the city's economic cycles. METHODS: An ecologic study based on the official death toll from violence registered between January 1997 and September 2006 in Santa Fe de Bogota, Colombia. Auto Regressive Integrated Moving Average (ARIMA) time series trimester models were used to describe the behavior of violent deaths. The explanatory variables for the city's economic cycles were the trimester series of the consumer price index, the unemployment rate, and the total number of people employed; these were related to the death series through transfer function models. RESULTS: Of the 36 575 total deaths from violence in Bogota, 57.7% were homicides and 23.7% resulted from traffic accidents. The homicide, suicide, and traffic accident deaths showed downward trends, with a noteworthy seasonal pattern in the traffic accident and suicide deaths that peaked during the last two trimesters of each year. Significant associations were found between all the death series from all the different causes and at least one economic series, and in each case a respective transfer function model was identified. CONCLUSIONS: Homicides constituted the largest portion of the violent deaths, and exposed a seasonal pattern to this death type. Significant associations were found between deaths from violence and some economic variables, revealing a cycle tied to the life of the city. The socioeconomic circumstances affecting a society were confirmed to influence the level of violence. PMID- 19814879 TI - [Infant mortality and urban marginalization: a spatial analysis of their relationship in a medium-sized city in northwest Mexico]. AB - OBJECTIVE: To identify areas with high risk of infant mortality and any possible correlation with the population's socioeconomic status through the use of a geographic information system and spacial analysis techniques. METHODS: An exploratory ecologic study was conducted in Hermosillo, the capital of Sonora, Mexico, in 2000-2003. The urban marginalization index (UMI) and the infant mortality rate (IMR) were determined for each of the city's basic geostatistical areas (BGA). The UMI and IMR were statistically calculated to identify geographic areas in which they were concentrated and to determine the degree of spatial correlation between these indicators. To determine the general spatial autocorrelation and spatial clustering of UMIs and IMRs within the city and the BGAs, Morans I index, Ipop statistics, and Besag and Newell's method were employed. RESULTS: The mean IMR was 14.3 per 1,000 live births, higher in the BGAs with greater social marginalization (16.2 per 1,000) and lower in those with less (11.7 per 1,000). The UMI range was -3.1-6.6 (maximum: 4.3; minimum: -2.7). Autocorrelation was found among the UMI (Moran I = 0.62), with significant clustering in the city's northwest, northeast, and southeast parts. Local clustering of high IMRs was found in Hermosillo's central and western areas, albeit without autocorrelation (Moran I = -0.007). High risk areas (high IMR and high UMI) were found in the city's northwestern section. CONCLUSIONS: Spatial clusters with high IMR were found in socially marginalized areas in the northwestern part of Hermosillo, a city of medium size located in northwestern Mexico. These results, reached through a combination of spatial analysis techniques and geographic information tools can help guide interventions specifically designed for these high risk residential areas. PMID- 19814880 TI - Chlamydia trachomatis infection and associated risk factors in a low-income marginalized urban population in coastal Peru. AB - OBJECTIVES: To estimate Chlamydia trachomatis (CT) infection prevalence and associated risk factors among a low-income marginalized urban population in Peru. METHODS: Between April 2003 and April 2005, men and women at high-risk for sexually transmitted infections (STIs) were recruited from low-income urban areas in three coastal cities in Peru (Chiclayo, Lima, and Trujillo). Consenting participants were studied using a sero-epidemiologic survey. Urine and vaginal swabs collected from men and women were evaluated using polymerase chain reaction (PCR) (COBAS AMPLICOR (CT/NG) Test, Roche Molecular Diagnostics, Branchburg, NJ, USA) for CT. RESULTS: Among the 2 440 participants recruited for the study (2 145 men and 295 women), overall prevalence of CT infection was 6.6% (95% CI, 5.6 7.6%): 5.5% (95% CI, 4.5-6.5%) in men and 14.9% (95% CI, 11.7-27.1%) in women. Chlamydial infection was inversely associated with age and positively associated with HIV infection and dysuria in men. Among women, chlamydial infection was inversely associated with age and positively associated with number of sex partners. CONCLUSIONS: CT infection was common among high-risk men and women in urban coastal Peru. Because chlamydial infection is associated with complications related to female reproduction, including infertility and ectopic pregnancy, interventions to prevent and treat infection and studies to determine the feasibility of population-based screening for CT should be conducted among the high-risk female population. PMID- 19814881 TI - [The pharmaceutical market in Mexico: size, value, and concentration]. AB - OBJECTIVES: To describe the pharmaceutical drug market in Mexico in terms of its size, structure, business' market power, and consumer negotiating power. METHODS: A descriptive study based on data from the 2004 Economics Census and the reports of IMS Health, Inc. (Norwalk, Connecticut, United States of America). Sales amounts and volumes of Mexico's pharmaceutical companies from 2002-2005 were obtained and the Herfindahl-Hirschman Index (HHI) and its inverse were calculated as indicators of the market's degree of concentration; also, price elasticity was determined by a product index. RESULTS: The total value of the products manufactured by the pharmaceutical sector was 115 billion in 2006 Mexican pesos, of which 99% pertained to companies categorized as large. This amount constituted 1.2% of the national gross domestic product that year (20.0% of the health sector's portion, estimated to be 6.0%) and 3.9% of the total value of manufactured goods. The HHI of Mexico's pharmaceutical market during the study period was about 0.04, albeit with a steady decline, and its inverse decreased from 23 to 26. The price elasticity of pharmaceutical products was minimal (0.007, 0.003, and -0.002). CONCLUSIONS: This study constitutes a preliminary description of Mexico's pharmaceutical market, one of the country's most dynamic economic sectors. It confirmed that the market is a rigid oligopoly, and thus supports enactment of firmer regulatory tools to reduce the power of the manufacturers in favor of that of the consumers. PMID- 19814882 TI - Accurately estimating breast cancer survival in Spain: cross-matching local cancer registries with the National Death Index. AB - OBJECTIVES: To assess the impact of using data from the National Death Index (NDI) of Spain to estimate breast cancer survival rates among residents of Girona and Zaragoza diagnosed in 1995-1999. METHODS: This was an observational, longitudinal epidemiologic study, using two population-based cancer registries. Data collected were of female residents of Girona or Zaragoza who had been diagnosed with breast cancer in 1995-1999. Observed and relative 5-year survival rates were estimated, first using the information available from the Girona and Zaragoza cancer registries, and then with the inclusion of NDI data. The 5-year relative survival rate and corresponding 95% Confidence Intervals were estimated using the Hakulinen method. The Kaplan-Maier method and Log Rank test were used to compare survival curves. RESULTS: No statistically significant difference in survival curves was observed in Girona for the data obtained before and after cross-matching with the NDI. However, there was a significant difference in Zaragoza. A comparison of the relative survival rates of each of the two registries before NDI cross-matching showed differences of 3.9% (5-year) and 16.1% (10-year) between the two, whereas after the cross-match, the difference was only 0.5% (5-year) and 1.2% (10-year). CONCLUSIONS: In Spain it is imperative that there be systematic use of NDI data to supplement cancer registries, so that comparisons of relative survival rates between registries can be improved. PMID- 19814883 TI - Cost-of-illness study of type 2 diabetes mellitus in Colombia. AB - OBJECTIVE: To determine the per patient and overall cost of illness of type 2 diabetes mellitus (T2DM) in Colombia from Ministry of Health and societal perspectives. METHODS: A published Markov transition model was adapted for Colombia, using the clinical expertise of a Colombian endocrinologist. Transition probabilities for the model were derived from an international literature review. A model was run for a time horizon of 42 years. Direct resources (drugs, laboratory, medical, hospital, other health care) were identified and cost was ascertained by using national price lists, international health care guidelines, and other Colombian studies or data from other countries. Indirect costs (work time lost) were calculated by using the human capital approach. Annual and lifetime direct and indirect costs, in 2007 U.S. dollars with a 5% discount rate, were determined on a per patient basis and projected to the overall Colombian population. Costs were clustered according to treatments and outcomes. RESULTS: The estimated annual cost was $2.7 billion from the societal perspective and $921 million from the Ministry of Health perspective. The annual direct cost per patient was $288, and the indirect cost was $559 (total = $847). This cost was distributed across disease outcomes as follows: diabetes treatment (drugs), 47%; cardiac and coronary disease, 24%; stroke, 15%; amputation, 9%; nephropathy, 3%; retinopathy, 2%. Macrovascular complications made up 86% of the annual direct costs and 95% of the annual indirect costs of T2DM. CONCLUSIONS: We estimated the annual cost of T2DM for Colombia from societal, Ministry of Health, and Colombian Health System perspectives. We also estimated annual direct cost per patient and the cost of treating diabetes and macrovascular complications. The economic burden is substantial and comparable to results for other countries. The model showed a logical disease progression. PMID- 19814884 TI - [Identifying areas of high risk for ophidism in Cordoba, Argentina, using SIGEpi software]. AB - OBJECTIVES: To determine the case distribution of accidental ophidism in the Cordoba province of Argentina, identify high risk areas, and evaluate the usefulness of the SIGEpi software program (Pan American Health Organization, Washington, DC, United States of America). METHODS: Information regarding the 299 cases of accidental ophidism reported in Cordoba in 1995-2006 was collected from the Provincial Epidemiology Department's official records of incidents involving venomous creatures. The SIGEpi software program was used for geographic mapping. Specific maps were produced to identify, quantify, and visualize the danger zones and areas of high risk for ophidism. RESULTS: The incidents occurred mostly in the northern and western areas of the province. Snakes of the Bothrops genus were responsible for the majority of the incidents (87.7% of the total). The departments with the highest annual incidence rates were Pocho and Rio Seco (> 10 incidents per 100,000 residents). Three ophidism high-risk zones were identified: one in far west of the province, another in the far north, and another just north of Banados del Rio Dulce and the Mar Chiquita lagoon. CONCLUSIONS: Accidental ophidism constitutes a major health problem in the Cordoba province where incidence rates are higher than those areas with more severe economic issues tied to rural labor. SIGEpi software is a reasonable choice for studying public health challenges and proved to be effective in identifying areas at high risk for ophidism. PMID- 19814885 TI - Depressive symptoms among poor older adults in Mexico: prevalence and associated factors. AB - OBJECTIVE: To describe the presence of depressive symptoms (DS) and factors associated with them among poor Mexican older male and female adults (OA). METHODS: A survey was conducted among OA enrolled in a governmental poverty relief program (Oportunidades) in Mexico. Participants completed a short version of the Center for Epidemiologic Studies Depression Scale and answered questions about health and life conditions. RESULTS: The prevalence of DS over the cutoff point was 43% (confidence interval (CI) 39.5-46.4). DS were associated with different variables among male and female participants. For men, difficulties in performing daily life activities were associated with DS over the cutoff point (odds ratio (OR) 2.62, CI 1.58-4.34) and literacy was associated with less DS (OR 0.46, CI 0.31-0.68). For women, difficulties with daily life activities were associated with more DS (OR 2.50, CI 1.57-3.97), and being head of the family was also positively associated (OR 1.81, CI 1.11-2.93). CONCLUSIONS: DS were frequent among this sample of poor OA. The differences between men and women in variables associated with DS highlight the importance of considering the gendered aspects of growing old. PMID- 19814886 TI - HIV/AIDS in Cuba: lessons and challenges. PMID- 19814887 TI - [Response to AIDS in Brazil: contributions of social movements and the sanitary reform]. AB - This paper briefly outlines how the political scenario and the mobilization of different actors have contributed to the construction of a public health policy in response to the AIDS epidemics in Brazil. Three factors are presented and discussed: the political context of the 1980s, characterized by redemocratization, growth of social movements, and consolidation of the Brazilian health care reform; the socio-cultural context of the 1970s and 1980s, characterized by achievement of individual freedom, which was key to the organization of the AIDS movement; and finally the actions carried out in the international scenario to support the sustainability of the Brazilian domestic policy and the reinforcement of a global response to face the epidemics in lower middle income economies. PMID- 19814888 TI - Cross-border utilization of health care services by United States residents living near the Mexican border. AB - OBJECTIVES: To determine what proportion of El Paso (Texas, United States of America) residents access health care services in Mexico, which services they use, and why they cross for care. METHODS: A cross-sectional, telephone survey of El Paso County residents was conducted from October-November 2007 to determine use of health care services in Mexico. At total of 2 560 telephone interviews were completed. Descriptive statistics and Chi-square analysis were used to determine the proportion crossing the US-Mexico border for care and identify correlates of crossing. RESULTS: The proportion of El Paso residents that had crossed into Mexico for some type of health care service during the two years prior to the survey interview was 32.5%. Of border crossers, 27.1% used health services; 63.2%, dental services; 82.0%, pharmacy; and 9.8%, traditional healers. Reasons given were cost, lack of health insurance, language barriers, and convenience. Hispanic ethnicity, having lived in Ciudad Juarez, being uninsured, and younger age were associated with crossing. The majority were satisfied with the health services received. CONCLUSIONS: Crossing the border to access health care services in Mexico is not uncommon for US residents living in El Paso County, Texas. Given the high rate of uninsured in the United States and the increasingly stringent security requirements, health policies may be needed that allow for cross-border care, making use of this alternative health care system easier and safer. PMID- 19814889 TI - [Direct medical costs of invasive pneumococcal disease and radiologically diagnosed pneumonia among Chilean children]. AB - OBJECTIVES: To determine the direct medical costs of health care services for cases of invasive pneumococcal disease (IPD) and pneumonia acquired in the community and confirmed by radiology (NAC-Rx) among Chilean children. METHODS: A prospective follow-up study of the health services delivered to 594 children 0-35 months of age with IPD and 1 489 children 1-35 months with NAC-Rx, diagnosed and treated by organizations within public health network of the Region Metropolitana de Chile. The value of the health services was established according to rates supplied by the Fondo Nacional de Salud (FONASA, the National Health Fund) and prices charged by two private clinics. The national IPD and NAC-Rx rates were estimated to calculate the total national economic burden for the population covered by state health insurance. RESULTS: The mean cost of cases requiring hospitalization was US$ 1 056.20 for IPD and US$ 594.80 for NAC-Rx, while that of cases treated by out-patient services was US$ 77.70 and US$ 65.20, respectively. The cost of the same services for in-patient care at the private clinics was US$ 4 484.10 and US$ 2 962.70 at one clinic and US$ 9 967.50 and US$ 6 578.40 at the other. The estimated national annual cost of services for children under 5 years of age, according to FONASA rates, was US$ 789 045 for IPD and US$ 13 823 289 for NAC-Rx. CONCLUSIONS: The high demand for services and financial resources for NAC Rx in children 0-3 years of age is a tremendously powerful public health reason to support the routine use of pneumococcal vaccination in Chilean children. PMID- 19814890 TI - Maternal low birth weight and adverse perinatal outcomes: the 1982 Pelotas Birth Cohort Study, Brazil. AB - OBJECTIVE: To explore the association between maternal low birth weight (LBW) and adverse perinatal outcomes and to discriminate between confounders and mediating factors of these associations in a population-based birth cohort of Southern Brazil. METHODS: Data from 794 female members of the 1982 Pelotas Birth Cohort Study known to have delivered a live-born singleton offspring up to December 2004 were analyzed. Maternal birth weights were recorded in 1982. The associations between maternal and offspring characteristics were estimated by Poisson regression. Confounding was tested for socioeconomic, demographic, and psychosocial factors. Maternal anthropometric characteristics and hypertensive diseases during pregnancy were considered mediating factors. RESULTS: An increase of 100 grams (g) in mothers' birth weight predicted a gain of 21 g in their infants' birth weight (95% confidence interval (CI) 13.0-29.0 g, P < 0.001). Maternal LBW was associated with offspring LBW (prevalence ratio (PR) 2.28 (95% CI 1.34-3.89), P = 0.002), preterm birth (PR 1.78 (95% CI 1.12-2.81), P = 0.01), and small for gestational age (SGA) (PR 1.93 (95% CI 1.14-3.26), P = 0.01). A causal chain linking maternal LBW and offspring SGA was mediated by maternal prepregnancy weight. CONCLUSION: Offspring of young women born with LBW are more likely to be LBW, preterm, and SGA. Public health strategies aimed at decreasing the frequency of LBW are necessary to reduce the perpetuation of adverse perinatal outcomes in later generations. The intermediate role of prepregnancy weight among LBW women opens a promising window to decreasing the prevalence of SGA in similar populations. PMID- 19814891 TI - Field and laboratory preparedness: challenges to rolling out new multidrug resistant tuberculosis diagnostics. AB - OBJECTIVES: In a pilot implementation project of the microscopic-observation drug susceptibility methodology, we conducted a process evaluation to identify health system and logistic challenges that need to be addressed in order to harness the benefits of rolling out promising new diagnostic tools for multidrug-resistant tuberculosis (MDRTB). METHODS: Regional data relating to health system practices and performance related to the MDRTB diagnostic algorithm were collected at health center, local, and regional laboratories. RESULTS: Parallel implementation of a new test and an existing method creates demands on funds, personnel, sample transport, and information systems in addition to new test startup costs. Obviating the need for primary culture at intermediate laboratories through direct drug susceptibility testing (DST) at the regional reference laboratory significantly reduces delay. Field application of well-defined national guidelines for DST is patchy. If fidelity to national guidelines were perfect, DST requests would increase more than 50-fold, with important implications for laboratory capacity. CONCLUSIONS: Implementing a new MDRTB diagnostic presents challenges to the laboratory environment, the existing DST process, and the application of national guidelines in peripheral clinics. Assessing each element can maximize efficient use of a new tool. Specifically, strengthening systems for transferring samples to the laboratory and delivering results to the requesting clinic in addition to investing in personnel and laboratory resources are integral to harnessing the benefits of high-performance new diagnostic tests and can bring added value to other programs in the health care system. PMID- 19814892 TI - [Violent youth gangs in Madrid: socialization and culturalization]. AB - OBJECTIVES: This study explores the subject of youth involved in violent groups or gangs, with the goal of further understanding the indoctrination, socialization, and culturalization processes undergone by youth involved in group violence or gangs. Furthermore, to examine the dynamics between peer pressure and other social factors (dating relationships, work, family, etc.) within the theoretical framework of the theories of primary socialization and differential socialization. METHODS: A qualitative analysis of 40 interviews of youth belonging to violent gangs/groups. RESULTS: According to the theories of primary socialization and differential socialization, over socialization by the violent group and under socialization by all other social entities can be assumed. Regarding parental supervision and support, three family types were clearly associated with the problem of youth violence. The distinct or unified social identity of the violent youth, as well as their individual self esteem and self image, formed a combination of processes whose relevance was highly predictive. Lastly, an accurate indicator of how these youth mature is their support network perceived, absolute, and relative (distributed among the various influencing forces). CONCLUSIONS: The study clearly outlines the need for re-imposing fundamental philosophical epistemology and methodologies on social forces of this kind, incorporating elements key to the postmodern, constructionist, and opposing perspectives. PMID- 19814893 TI - Proposed methodology for monitoring antiretroviral drugs price negotiations in Latin America and the Caribbean. AB - OBJECTIVES: The spread of HIV/AIDS challenges governments to provide antiretroviral (ARV) treatment at affordable prices, and various initiatives have been developed with that intent. In Latin America and the Caribbean, four subregional negotiations were conducted during 2002-2005 to reduce drug prices and thus broaden access to ARVs. Studies were carried out to monitor the negotiations, and the development of a monitoring methodology was recommended. The objective of the current study was to develop and describe a potential methodology for monitoring ARV price negotiations. METHODS: The study, carried out in 2006-2007, consisted of a design phase and validation phase. The design phase included an extensive literature review and development of a theoretical framework. Validation was performed using health professional consensus and pilot studies in three countries-Barbados, Honduras, and Peru-representing the Caribbean, Central American, and Andean subregions. RESULTS: The results included a detailed logic model and a 40-indicator framework. Both were tested in the field. Indicators were evaluated for feasibility, pertinence, and sensitivity, based on the outcome of the pilot study. CONCLUSIONS: This monitoring methodology is designed to help countries self-evaluate progress toward implementation of ARV price negotiations. The results of the pilot study indicate that its implementation in the field helped elucidate the ARV price negotiation process by identifying local conditions and indirectly measuring countries' negotiating capacities. PMID- 19814894 TI - Assistive technology applied to education of students with visual impairment. AB - OBJECTIVES: Verify the application of assistive technology, especially information technology in the education of blind and low-vision students from the perceptions of their teachers. METHODS: Descriptive survey study in public schools in three municipalities of the state of Sao Paulo, Brazil. The sample comprised 134 teachers. RESULTS: According to the teachers' opinions, there are differences in the specificities and applicability of assistive technology for blind and low-vision students, for whom specific computer programs are important. Information technology enhances reading and writing skills, as well as communication with the world on an equal basis, thereby improving quality of life and facilitating the learning process. The main reason for not using information technology is the lack of planning courses. The main requirements for the use of information technology in schools are enough computers for all students, advisers to help teachers, and pedagogical support. CONCLUSIONS: Assistive technology is applied to education of students with visual impairment; however, teachers indicate the need for infrastructure and pedagogical support. Information technology is an important tool in the inclusion process and can promote independence and autonomy of students with visual impairment. PMID- 19814895 TI - Assessment of the quality of immunization data produced by the national individual registration system in Uruguay, 2006. AB - OBJECTIVE: The nominal registration system of Uruguay's national immunization program (NIP) tracks administered vaccines on a paper form filled out after each vaccination and collated into a national database, thus allowing for individual follow-up. This study performed a comprehensive assessment of the quality of Uruguay's immunization data in November 2006 to evaluate the validity of the information and to confirm the high national immunization coverage reported by the program. METHODS: The research team analyzed the concordance of the operational-level numerators (infant immunization data from 18 public and private vaccination centers in six country departments) with department- and national level data, and compared the national-level (NIP) infant denominators with other official sources. A standardized questionnaire was used to evaluate system performance at the operational (vaccination center), department, and national level. Rapid house-to-house monitoring was conducted to generate additional coverage estimates. RESULTS: Numerator accuracy throughout the data flow was 100%, and national-level denominators appeared to be exhaustive. Overall system performance was excellent (proper archiving and recording of form data, sufficient supply of forms, timely flow of information, adequate defaulter tracing practices and computer system security). The main weaknesses were the degree of data analysis and feedback to peripheral levels. House-to-house monitoring showed high overall immunization coverage (97%). CONCLUSION: Uruguay's NIP registration system produces remarkably reliable information, ensuring valid measurement of immunization coverage. In addition, by allowing for monitoring of each child's current vaccination status, it facilitates management of interventions designed to reduce vaccination default and thus helps achieve the country's high level of coverage. PMID- 19814896 TI - [Economic factors related to the Millennium Development Goals: a literature review]. AB - OBJECTIVES: To systematize all the information published on the status of and progress made toward the Health-related Millennium Development Goals (HMDGs), as well as to understand associations with certain economic factors and the potential for success. METHODS: A search was conducted for all scientific articles covering the Millennium Development Goals in general, published from 1 January 2000 to 31 August 2006, in the electronic databases of the EBSCO, CSA Illumina, Thomson Gale, SwetsWise, and BIREME. All original articles in English or Spanish that evaluated HMDG status, progress, and determinants were selected. The analysis evaluated the distribution of determinants of HMDG status or progress, the HMDGs referred to, the study type, the relationship between economic indicators and health, the study location, and the status and potential for attaining the HMDGs. The quality of the articles was also rated. RESULTS: Of the 304 original articles found, 114 (37.5%) covered one or more HMDGs. The most frequently addressed goals were those concerning infant and maternal mortality. Of the 39 articles that evaluated HMDGs and their association with economic variables, 13 dealt with economic factors related to equity, policy, or globalization. Economic and policy factors were most frequently associated with HMDG status or progress. CONCLUSIONS: There is a definitive trend toward measuring HMDG status and progress according to economic factors that reflect the population's condition. There is an information gap regarding government spending, international commerce, international aid, and global economic policy. The potential for achieving HMDGs in poor countries is low. PMID- 19814897 TI - [New self-care technologies and the risk of self-diagnosis through the Internet]. AB - Encouraged by a solid commercial structure and growing cultural influence, information and communication technologies for health (ICTH) appear as a natural reference point for matters of self-care. As a first trend, ICTH involved end user consultations to websites specializing in health. Soon after, the development of algorithms focused on the identification of illnesses to meet the needs of consumers seeking distant, impersonal technical advice. Despite the comfort and confidentiality, there is evidence that such resources have generated more doubt and anxiety than meaningful clarifications. According to the literature, users seem to have difficulties in identifying and naming their own symptoms, in addition to having to choose between shallow or alarmist advice. Despite the investments made in providing information, most sources of information do not rely on previous cultural studies about how users interpret their health conditions when resorting to self-diagnosis, which ends up by pushing them, paradoxically, to look for specialists. PMID- 19814898 TI - [Development of primary health care competencies]. AB - Given that the lack of qualified human resources devoted to primary health care (PHC) is one of the reasons why Alma Ata goals are as yet unattained, PAHO/WHO sponsored the design and implementation of a virtual course for training health care professionals at this level. The course was designed around competencies identified as being necessary for renewing primary care, founded on the premise that providing quality, sustainable services amenable to the public must emanate from professionals who lead the change process at all levels in the system. In this first phase, instruction is aimed at PHC leaders, managers, and other decision makers. The course is offered through the Virtual Public Health Campus and is 27 weeks long. PMID- 19814899 TI - [National public health institutes: a coordinated and comprehensive approach toward influenza A H1N1 and other threats to public health]. AB - The current influenza A H1N1 epidemic has demonstrated once again the importance of being able to count on robust, coordinated, and comprehensive public health systems. Countries that do rely on such health systems have proven to recognize, diagnose, and treat influenza in a timely manner, and to provide the public with the education needed to minimize the number of deaths and acute cases. The International Association of National Public Health Institutes (IANPHI) recommends that all countries begin to coordinate their national public health efforts. IANPHI offers its support and guidance to all who would create or strengthen their national public health institutes. By strengthening national public health systems, the ability to collaborate and the security of all countries is heightened. These institutions exist for the public good, and any improvements made to them are also a contribution toward better health for the population. PMID- 19814900 TI - Special issue for Terry Caelli. PMID- 19814901 TI - Coupling the world with the observer: from analysis of information to active vision. AB - In this paper we define the content of information in an image and show how it can be computed by taking into account different levels of resolution, in the framework of information theory and the thermodynamics of irreversible transformations. The results thus obtained will eventually be exploited to derive a mechanism for active exploration of visual space suitable to perform a dynamic coupling between the agent and its environment. PMID- 19814902 TI - Contrast- and illumination-invariant object recognition from active sensation. AB - It has been suggested that the deleterious effect of contrast reversal on visual recognition is unique to faces, not objects. Here we show from priming, supervised category learning, and generalization that there is no such thing as general invariance of recognition of non-face objects against contrast reversal and, likewise, changes in direction of illumination. However, when recognition varies with rendering conditions, invariance may be restored and effects of continuous learning may be reduced by providing prior object knowledge from active sensation. Our findings suggest that the degree of contrast invariance achieved reflects functional characteristics of object representations learned in a task-dependent fashion. PMID- 19814903 TI - Efficient visual coding and the predictability of eye movements on natural movies. AB - We deal with the analysis of eye movements made on natural movies in free-viewing conditions. Saccades are detected and used to label two classes of movie patches as attended and non-attended. Machine learning techniques are then used to determine how well the two classes can be separated, i.e., how predictable saccade targets are. Although very simple saliency measures are used and then averaged to obtain just one average value per scale, the two classes can be separated with an ROC score of around 0.7, which is higher than previously reported results. Moreover, predictability is analysed for different representations to obtain indirect evidence for the likelihood of a particular representation. It is shown that the predictability correlates with the local intrinsic dimension in a movie. PMID- 19814904 TI - Representation of space: image-like or sensorimotor? AB - We investigate the relation between the physical world and its mental representation in the 'cognitive map', and test if this representation is image like and complies with the laws of Euclidean geometry. We have developed a new experimental technique using 'impossible' virtual environments (VE) to directly influence the representational development. Subjects explore a number of VEs -- some 'normal', others with severe violations of Euclidean metrics or planar topology. We check if these manipulated properties cause problems in navigation performance. A consistent VE should be easily represented mentally in a map-like fashion, while a VE with severe violations should prove difficult. Surprisingly, we found no substantial influence of the impossible VEs on navigation performance, and forced-choice tests showed little evidence that subjects were aware of manipulations. This suggests that the representation does not resemble a two-dimensional image-like map. Alternatives to consider are sensorimotor and graph-like representations. PMID- 19814905 TI - Novel kernels for error-tolerant graph classification. AB - One of the major difficulties in graph classification is the lack of mathematical structure in the space of graphs. The use of kernel machines allows us to overcome this fundamental limitation in an elegant manner by addressing the pattern recognition problem in an implicitly existing feature vector space instead of the original space of graphs. In this paper we propose three novel error-tolerant graph kernels -- a diffusion kernel, a convolution kernel, and a random walk kernel. The kernels are closely related to one of the most flexible graph matching methods, graph edit distance. Consequently, our kernels are applicable to virtually any kind of graph. They also show a high degree of robustness against various types of distortion. In an experimental evaluation involving the classification of line drawings, images, diatoms, fingerprints, and molecules, we demonstrate the superior performance of the proposed kernels in conjunction with support vector machines over a standard nearest-neighbor reference method and several other graph kernels including a standard random walk kernel. PMID- 19814906 TI - Faster graphical models for point-pattern matching. AB - It has been shown that isometric matching problems can be solved exactly in polynomial time, by means of a Junction Tree with small maximal clique size. Recently, an iterative algorithm was presented which converges to the same solution an order of magnitude faster. Here, we build on both of these ideas to produce an algorithm with the same asymptotic running time as the iterative solution, but which requires only a single iteration of belief propagation. Thus our algorithm is much faster in practice, while maintaining similar error rates. PMID- 19814907 TI - Spatial-temporal modeling of interactive image interpretation. AB - We consider the problem of spatial-temporal modeling of interactive image interpretation. The interactive process is composed of a sequential prediction step and a change detection step. Combining the two steps leads to a semi automatic predictor that can be applied to a time-series, yields good predictions, and requests new human input when a change point is detected. The model can effectively capture changes of image features and gradually adapts to them. We propose an online framework that naturally addresses these problems in a unified manner. Our empirical study with a synthetic data set and a road tracking dataset demonstrate the efficiency of the proposed approach. PMID- 19814913 TI - Home and school environmental assessment and remediation. AB - Children spend a considerable portion of their time indoors. Therefore, homes and schools are an important source of allergen exposure. Chronic exposure to the major indoor allergens can lead to allergic sensitization and provoke allergic symptoms in children. Environmental assessment is crucial for the identification and quantification of such allergens in indoor spaces. Reduction of allergen exposure below sensitization and symptom thresholds is possible with various remediation techniques. This article reviews and discusses evidence for the assessment and remediation of indoor allergens commonly found in homes and schools. A literature review was performed using the PubMed database for English language articles published between January 1, 1980, and February 2009. Additional information was obtained from a review of recent textbooks and one professional society's webpage. PMID- 19814914 TI - Personalized medicine: a pediatric perspective. AB - The aim of pediatric personalized medicine is to uniquely combine genetic variation with developmental stage and environmental exposure to provide a tailored preventive, diagnostic, and therapeutic regimen. Recent advances in genomic research have identified many genetic variants that may be related to allergic and inflammatory disease and therapeutic response. These include variants involved in immune response, barrier proteins, and medication response. Current evidence also suggests that the effect of genetic variation often depends on the developmental stage of a child and environmental exposure such as infection or tobacco smoke during a specific stage. Personalized medicine is a new and exciting field with the potential to significantly improve medical care for children and adults. PMID- 19814915 TI - An update on exercise-induced bronchoconstriction with and without asthma. AB - Exercise-induced bronchoconstriction (EIB) is defined as transient, reversible bronchoconstriction that develops after strenuous exercise. It is a heterogeneous syndrome made up of a spectrum of phenotypes ranging from the asymptomatic military recruit whose condition is detected by diagnostic exercise challenge to the athlete with known asthma to the elite athlete for whom EIB represents an overuse or injury syndrome. If exercise is the only identified trigger for bronchoconstriction, it is called EIB. However, when it is associated with known asthma, then it is defined as EIB with asthma. This review discusses the pathogenesis, presentation, diagnosis, and management of EIB and EIB with asthma. PMID- 19814916 TI - Asthma phenotypes. AB - The many roads leading to the syndrome of asthma have proven to be intricately interconnected. The chronic inflammation of asthma is characterized by airway hyperreactivity and variable reversibility. Past classification systems relied on assessment of daily impairment and the distinction between intrinsic (nonallergic) and extrinsic (allergic). With more precise asthma phenotypes, association studies likely will have greater significance. In addition, patients at higher risk for severe disease can be more effectively managed, and treatments can be directed to responders. In this review, we look at eight identified phenotypes: atopic and nonatopic status, pre-asthma wheezing, inflammatory mediator predominance, aspirin-sensitive, exercise-induced, severe, and flare prone asthma. Although significant overlap exists among the distinctions, any future phenotype classification system will need to incorporate these eight clinical asthmatic populations. PMID- 19814918 TI - Otitis media and antihistamines. AB - Otitis media with effusion (OME) and acute otitis media (AOM) continue to be a significant source of morbidity in the United States, as they account for more than 3.5 billion dollars in annual costs, and OME is the sixth most common reason for an emergency department visit. The efficacy of antihistamines in the treatment of OME/AOM has been thoroughly refuted in the literature during the past three decades. However, most studies to date have used first-generation antihistamines with or without decongestants. We propose that second-generation antihistamines may have increased efficacy in the treatment of OME/AOM because of their greater selectivity for histamine receptors and lack of anticholinergic activity. Further clinical trials may be warranted, as medical treatment with second-generation antihistamines, if proven more efficacious, may reduce the need for antibiotic treatment and surgical intervention. PMID- 19814917 TI - Biofilms in pediatric respiratory and related infections. AB - Bacteria can grow as free-floating, planktonic bacteria or complex communities called biofilms. Biofilms promote bacterial growth and diversity and offer bacteria unique environments, including aerobic and anaerobic layers, that facilitate resistance to antimicrobial therapies. Respiratory and related structures provide ideal environments for the development of bacterial biofilms, which predispose patients to recurrent and chronic infections. Biofilms are important for the persistence of chronic rhinosinusitis, pulmonary infections in cystic fibrosis, chronic otitis media, and device-related infections. Antimicrobial therapy that is proven effective against planktonic bacteria is often insufficiently effective against the defenses of biofilms. Furthermore, biofilms modify themselves following exposure to antimicrobial therapy, thus developing increased resistance. Understanding the nature of biofilms in common pediatric infections is essential to comprehending the expected course of bacterial illness and identifying treatments that are most likely to be beneficial against more resistant biofilms. PMID- 19814919 TI - Role of adenoids and adenoiditis in children with allergy and otitis media. AB - Adenoids and/or tonsil inflammation with concomitant obstructive hypertrophy is one of the oldest and most common pediatric problems. Adenoids are a component of Waldeyer's ring and because of their anatomic position can be relevant in the pathogenesis of otitis media when they are inflamed and/or enlarged. Adenoid pads can create mechanical eustachian tube obstruction. Therefore, in some cases, adenoidectomy may have a role in the clinical management of otitis media with effusion. However, eustachian tube dysfunction related to the adenoids may also have an allergy-related functional component. Allergic inflammation has been described for middle ear effusion, and some studies have reported that mast cells increase and allergic mediators release in adenoids as well. Nasal endoscopy has a key role in confirming a diagnosis of adenoid hypertrophy and/or adenoiditis and in detecting an association between adenoid inflammation/infection and otitis media with effusion, especially during infancy and early childhood. PMID- 19814920 TI - Relationship between pediatric obesity and otitis media with effusion. AB - Although eustachian tube dysfunction and bacterial infection have been shown to cause otitis media with effusion (OME), other etiologies are possible. One of the most common medical conditions in children is obesity, which can have effects throughout the body. Little is known, however, about the relationship between obesity and OME. Obesity may result in altered cytokine expression, gastroesophageal reflux disease, or fat accumulation, all of which may contribute to OME. Conversely, OME may induce taste changes through middle ear cavity inflammation, thus contributing to obesity. A similar pattern of taste change has been shown in patients with gustatory nerve anesthesia. Further research on the relationship between obesity and OME may help to determine the exact etiology of OME and contribute to our knowledge about the causes of obesity. PMID- 19814921 TI - Immunomodulators in asthma therapy. AB - New developments in the field of allergy and immunology have yielded a variety of novel therapeutic approaches in recent years, and more agents are at the clinical trial stage. Among the therapeutic approaches discussed in this review are Toll like receptor agonists, immunostimulatory oligodeoxynucleotides, orally and parenterally administered cytokine blockers, and specific cytokine receptor antagonists. Transcription factor modulators targeting syk kinase, peroxisome proliferator-activated receptor-gamma, and nuclear factor-kappaB are also being evaluated in the treatment of asthma. The anti-IgE monoclonal antibody omalizumab has established effectiveness in patients with allergic asthma, but the criteria for selecting patients who are most likely to benefit from it are less clear. This review summarizes data from human clinical trials with immunomodulators to discuss the rationale for their use, their efficacy, and adverse events associated with them. PMID- 19814922 TI - Neural control of airway inflammation. AB - Abnormal neural function contributes to the pathogenesis of airway disease. In addition to affecting airway physiology, the nerves produce and release inflammatory mediators, contributing to the recruitment and activation of leukocytes. Activated inflammatory cells in turn affect the function of airway nerves, changing the production and release of neurotransmitters. Cross-talk between airway nerves and leukocytes helps to maintain chronic inflammation and accentuates neural control of the airways. PMID- 19814923 TI - [Is there a future for medical treatment of obesity?]. PMID- 19814924 TI - [Laparoscopic surgery for colorectal cancer in Denmark]. AB - INTRODUCTION: Laparoscopic resection of colorectal cancer may be oncologically equal to open surgery. Since 2001 the departments performing colorectal cancer surgery in Denmark, have reported variables for patient diagnostics, treatment and course to the DCCG database. MATERIAL AND METHODS: This article is based on data for patients classified with laparoscopic procedure until 31.12.2007. RESULTS: From an annual number of 50 operations, the number increased to 761 completed laparoscopic operations in 2007. The median number of investigated lymph nodes in radical colon operation increased from 11 in 2001-2005 to 15 in 2007. The median blood loss was 50 ml. The postoperative complications were 27% after rectum surgery and 20% after colon surgery, whereas the rate of anastomotic leakage was 7% in both groups. Total 30-day-mortality was 5.5% in the rectum group and 3% in the colon group, and the median length of stay was eight and four days, respectively. Departments performing more than 100 laparoscopic procedures removed significantly more lymph nodes during colon surgery and had lower rate of anastomosis leakage compared with departments with less experience. CONCLUSION: The oncologic quality of laparoscopic surgery was satisfying in the short term. The postoperative complication rate was equal to international results. Yet, a rate of 7% anastomotic leakage after colon surgery warrants further investigation. PMID- 19814925 TI - [Penetrating trauma treated by the Medical Emergency Care Unit in Copenhagen]. AB - INTRODUCTION: The World Health Organization defines violence as a public health problem, affecting mainly boys and young men. Interpersonal violence with sharp objects and arms is rare in Denmark, but has been the focus of attention from the media and general population. This study describes the incidence and characteristics of penetrating trauma treated by the Medical Emergency Care Unit in Copenhagen (MECU) from 2000 to 2007. MATERIAL AND METHODS: The study is a historical cohort study with 30-day follow-up. Patients treated by the MECU for penetrating trauma were included. The physician manning the MECU prospectively registered clinical data and pre-hospital treatment for later analyses. Mortality after 30 days was assessed via the Danish Civil Registry System. RESULTS: A total of 764 patients were eligible for inclusion. We were unable to identify three patients due to missing civil registration numbers. Among the remaining 761 patients, 86% were male with a median age of 32 years. A total of 12 foreigners could only be followed to hospital discharge. The annual incidence of penetrating trauma remained unchanged around 93 cases throughout the study period (p = 0.39). The percentages of trauma caused by violence, as well as 30-day mortality, were constant. CONCLUSION: The incidence of and mortality from penetrating trauma has not been rising in Copenhagen during the last eight years. PMID- 19814926 TI - [Use of plastic adhesive drapes during surgery may increase the risk of surgical site infections. A survey of a Cochrane review]. AB - In theory, the products act as a barrier, which hinders the spreading of bacteria from the deeper skin layers and hair follicles to the incision. On the other hand, the use of plastic adhesive drapes may promote bacterial overgrowth due to a >>greenhouse effect<<. This Cochrane review which is based on seven trials showed that there was no evidence that plastic adhesive drapes reduces the surgical site infection rate and some evidence that they increase infection rates in clean operations. Consequently, their use should be abandoned. Further studies are warranted to determine the effect of other adhesive products currently used. PMID- 19814927 TI - [Dopamine agonist therapy in early Parkinson's disease. A survey of a Cochrane review]. AB - The Cochrane analysis confirms some well-known facts: initial therapy with dopamine agonists (DA) delays the incidence of motor complications, but is associated with inferior motor control and a higher frequency of non-motor adverse events (AE) compared with levodopa. Some serious AE are missing, probably due to the design of the included studies. The results will not change current treatment guidelines, but they emphasize the need for >>real-life<< studies with patients' quality of life or economic parameters as primary outcomes. PMID- 19814928 TI - [Benzodiazepine reduction does not imply an increased consumption of antidepressants. A survey of two medical practices]. AB - INTRODUCTION: The consumption of benzodiazepines (BZ) and cyclopyrrolones (CP) has attracted great interest in recent years due to the serious side-effects associated with these drugs. Two medical practices introduced a more restrictive approach to the prescription of BZ and CP. The intervention reduced the total consumption of anxiolytica by 75% and hypnotica by 90% during the two and a half year period. There is a general lack of knowledge about whether an intervention of this nature reduces drug consumption in general, or merely substitutes the consumption of drugs. Here, antidepressants (AD) are especially relevant substitutes. MATERIAL AND METHODS: In two medical practices, the consumption of AD was followed before, during and after the intervention. Consumption was followed via the Danish Medicines Agency's website Ordiprax, which listed the amount of prescription medicines sold from pharmacies. RESULTS: The total quantity of prescribed AD remained unchanged in the two practices (p = 0.05). There were minor shifts in prescription classes. This result should be seen in the context of the consumption of AD in the county as a whole, which rose by 8.6% per annum during the intervention period. CONCLUSION: In the primary sector, it is possible to reduce the amount of BZ and CP without causing a concomitant increase in the prescription of AD. The two medical practices of the present survey did not introduce a deliberately restrictive approach, but simply an increasing awareness when prescribing to patients, especially regarding the duration of prescriptions. PMID- 19814929 TI - [Management of serum calcium with cinacelet in parathyroid cancer]. AB - Treatment with cinacalcet and intravenous bisphosphonate has improved the previously often fatal prognosis of parathyroid cancer. We report two cases, of which one did not react to treatment and died within a short time-frame. In the other case, cinacalcet controlled serum calcium during a prolonged time period despite metastases. However, increasing doses of cinacalcet combined with more frequent intravenous bisphosphonate were necessary. PMID- 19814930 TI - [Fatigue fractures of the hip in healthy marathon runner]. AB - Fatigue fractures of the hip are rare causes of hip pain. Delayed diagnostics of this type of fracture can cause serious complications. We describe a case concerning a 32-year-old male amateur runner who experienced hip pain with rapid onset during his first marathon. After eight days his general practitioner referred him to the hospital and a dislocated fracture of the femoral neck was detected. PMID- 19814931 TI - [Mastitis-like changes as debut symptom of Kaposi's sarcoma and AIDS]. AB - Kaposi's sarcoma of the breast is rare but has been reported to imitate breast lumps or gynaecomastia, usually accompanied by Kaposi's sarcoma of the skin. We report a case of a patient presenting with mastitis of the breast as the first symptom of Kaposi's sarcoma and AIDS. Although uncommon, the diagnosis should be considered in patients with mastitis without other obvious causes. PMID- 19814932 TI - [Convalescence and sick leave--evidence versus tradition]. PMID- 19814933 TI - [Convalescence, sick leave and health economics]. AB - National health policies have been proposed to reduce long-term absenteeism in order to increase labour supply. Convalescence interventions have been developed and optimised and shown to reduce long-term absenteeism and as such they form an integral part of the national health policy. We describe absenteeism and convalescence within an economic perspective. In Denmark, the economic costs of absenteeism amounted to more than 37 billion Danish kroner in 2006, including sick leave benefits and non-productive wage expenditure. Optimising convalescence interventions, if efficacious, is highly cost-effective. PMID- 19814934 TI - [Recovalescence and sick leave following total knee and hip arthroplasty]. AB - Length of stay has been decreasing after total knee and hip arthroplasty (TKA and THA) in Denmark during recent years, and patients regain functional capacity faster - and it is therefore of importance to reevaluate the sick-leave period and recommendations given. The factors potentially influencing recovalescence are limited to pain, swelling, reduced range of motion, postoperative nausea and vomiting and sleep disturbances. This article evaluates these changes and gives recommendations with regard to sick leave and restrictions after operation. PMID- 19814935 TI - [Reconvalescence after surgery for proximal femoral fractures]. AB - In Denmark, 10,000 patients with hip fractures are annually admitted to hospital. These patients are generally old with a high degree of comorbidities. Current treatment strategies for peri- and postoperative treatment are presented in relation to patient reconvalescence. Overall expected hospitalization is 5-25 days with 1-3 months of analgesic treatment and physiotherapy. The 30-day mortality is 10% and the reoperation rate is 10-20%. The use of specialized centres and accelerated clinical programmes in Denmark is recommended. PMID- 19814936 TI - [Convalescence and sick leave following inguinal hernia repair]. AB - The duration of convalescence after inguinal hernia repair is determined by a number of factors, among which are information and recommendations given by the surgeon and the general practitioner, level and duration of postoperative pain, type of repair, level of physical activity at work and during leisure activities and development of complications. The literature provides no evidence to support an increased risk of recurrence with early resumption of physical activity after inguinal hernia repair. Patients should be encouraged to resume full activities immediately after surgery, only limited by pain. PMID- 19814937 TI - [Reconvalescence following Caesarean section]. AB - Recommendations for reconvalescence and restriction of physical activity after elective caesarean section are not consistent and there is a lack of evidence on the subject. Although physiological changes are rapidly normalized after the operation, the women experience an extended period characterized by fatigue caused by lack of sleep due to nursing and breastfeeding of the baby. This paper accounts for the effects of post-operative pain, breastfeeding, fatigue, sexuality and physical recovery in the period of reconvalecence following elective caesarean section. PMID- 19814938 TI - [Convalescence and sick leave following transurethral prostatectomy]. AB - Recovery after transurethral prostatectomy (TUR-P) is characterized by lower urinary tract symptoms (LUTS) and haematuria often affecting the patient's social life negatively. Procedure-specific information reduces the patient's anxiety postoperatively. When giving advice on level of activity during recovery, the risk of haematuria is the most important factor. Since bleeding ceases in 95% of cases within three weeks, patients should avoid hard physical activity for three weeks. Normal activity can be resumed immediately, although bothersome LUTS may be a limiting factor. Equivalent advice should be given with regard to the need for sick-leave. PMID- 19814939 TI - [Convalescence and sick leave after colonic surgery]. AB - The benefits of fast-track surgery are improved recovery and a shorter period of postoperative convalescence. After uncomplicated colonic resection, there is no pathophysiological basis for recommending a postoperative convalescence of more than 2-3 weeks. Advice after discharge - Convalescence: Daily activities and light exercise should be resumed as soon as possible. Physically moderate work should be resumed after two weeks. Physically severe exercise and lifts (> 10 kg) as well as physically strenuous work may be resumed after 3-4 weeks. PMID- 19814940 TI - [Convalescence and sick leave following cardiac surgery]. AB - A literature review regarding symptoms and recommendations in relation to e.g. driving and physical activity precautions during the recovery period following coronary artery bypass and/or valve surgery was performed. Postoperative pain, fatigue, anxiety, depression, sleeping problems and neurocognitive impairment affecting daily living are common. Every patient should be offered individually tailored cardiac rehabilitation. Patients should be on sick leave for 4-8 weeks following sternotomy. Recommendations during convalescence are based on a low level of evidence. PMID- 19814941 TI - [Convalescence and sick leave after influenza]. AB - Seasonal influenza has a significant impact on individuals and society alike. In otherwise healthy adults, a typical case of seasonal influenza is associated with six to eight days of clinical symptoms, and about four to five days of sick leave. Transmission mainly takes place during the initial four days of the illness. Convalescence from disease should be expected to comprise seven days from symptom onset. In mild cases with symptoms lasting only few days, convalescence could be limited to four days unless prevention of transmission is given very high priority. Shorter duration of convalescence and workplace absence is not recommended due to risk of transmission. PMID- 19814942 TI - [Convalescence and decline in physical function level following intensive therapy]. AB - More patients survive critical illness, which emphasises the need to assess outcome measures other than mortality. A prolonged decline in physical function is frequently observed after discharge in the critically ill. Neuromuscular dysfunction and muscle atrophy incurred during intensive care may prolong convalescence after discharge. Thus, strategies to counteract neuromuscular dysfunction and to improve physical outcome may reduce the overall burden of critical illness. This review describes the most common predisposing factors and discusses preventative measures and interventions. PMID- 19814943 TI - [Elective surgery reconvalescence after pre-operative physical training]. AB - Elective surgery is associated with decreased postoperative musculoskeletal function. High aerobic capacity and muscle strength prior to operation is associated with a shorter and more uncomplicated postoperative period. Pre operative training as intervention has only showed very limited effect in relation to abdominal or alloplastic surgery, whereas strength and coordination training prior to operation has been shown to have a beneficial effect on postoperative movement function after reconstructive ligament surgery of the knee. PMID- 19814944 TI - [A Danish intestinal pseudomyiasis case caused by Eristalis tenax]. AB - Intestinal pseudomyiasis caused by the larvae of the drone fly Eristalis tenax is sporadically reported and symptoms are varying. We report a ten-year-old boy with intermittent nonspecific abdominal pain. He noticed a larva in his stools which was later identified as the rat-tailed larva of Eristalis tenax. After passing the larva his symptoms subsided. No treatment was given. We aim to register the first case of human pseudomyaisis caused by E. tenax in Denmark. PMID- 19814945 TI - [Foot- and ankle trauma in a diabetic patient]. AB - If confronted with a neuropathic patient with an acute foot-ankle trauma, including fracture, this patient is by definition Eichenholtz stage 0. This is of major importance for the treatment. Whereas the surgical handling of fractures does not differ from ordinary fracture treatment, the subsequent treatment of sprains and fractures with immobilisation and a non-weight bearing period is significantly prolonged, frequently doubled, in diabetic patients. Follow-up includes frequent ambulatory controls including X-rays. If the neuroarthropathy progresses, the period of treatment is adjusted accordingly. PMID- 19814946 TI - [Hungry bone syndrome due to vitamin-D deficiency]. AB - An 85-year-old woman was admitted after being bedridden for six months with muscular weakness and diarrhoea. Vitamin 25-OH-D, serum calcium, magnesium and phosphate were low, and parathyroid hormone as well as alkaline phosphatase were increased. Dual x-ray absorption scan showed reduced bone mineral density. The patient was treated with calcium, magnesium and vitamin D, initially intravenously and subsequently as tablets. Clinically the patient's condition slowly normalized. Remineralization of the skeleton resulted in consistent hypocalcaemia for more than nine months. PMID- 19814947 TI - Effects of olanzapine and risperidone on glucose metabolism and insulin sensitivity in chronic schizophrenic patients with long-term antipsychotic treatment: a randomized 5-month study. AB - BACKGROUND: Comparisons of diabetic potential, glucose related metabolic levels, and insulin resistance between olanzapine and risperidone have produced variable results in cross-sectional and epidemiologic studies. Randomized prospective studies of metabolic effects during treatment with these drugs may provide results that are more informative. METHOD: Hospitalized patients with chronic schizophrenia (DSM-IV), most of whom had been treated with multiple antipsychotics in the past, were randomly assigned to treatment with a single antipsychotic, olanzapine or risperidone, for a period of 5 months. At baseline and every treatment month thereafter, fasting glucose, insulin, insulin-related metabolic measures, and prolactin were assessed, and an oral glucose tolerance test (OGTT) was performed during baseline and months 1, 2, and 5 of treatment. Weight was assessed monthly, and waist and hip measures were taken at baseline and month 5. Data were analyzed on 23 patients randomly assigned to risperidone and 23 patients randomly assigned to olanzapine. The study was conducted from February 2003 to August 2007. RESULTS: Most patients were overweight or obese at baseline (mean body mass index [BMI] = 29.4), but there were no differential drug effects on weight change and no differences between drug groups at the 5-month time point. There were no overall drug treatment differences in fasting glucose or glycohemoglobin or 2-hour glucose levels in OGTT and no differences between the two drug groups at the 5-month time point. There were no consistent drug treatment differences in the number of patients who developed borderline or diabetic glucose levels. Olanzapine-treated patients showed a significantly greater increase than risperidone-treated patients in a fasting measure of insulin resistance (P = .041), and olanzapine patients showed greater decreases in insulin sensitivity during OGTT (P = .023) compared to risperidone-treated patients. Olanzapine-treated patients had a significantly greater increase in 1 hour glucose and insulin levels during OGTT in subsequent months compared to baseline and greater increase in glucose and insulin area under the curve over time than the risperdone-treated patients. Prolactin levels decreased in olanzapine patients and increased in risperidone patients (P values approximately .02). There were no significant drug treatment differences in C-peptide levels or 2 indices proposed as measures of insulin secretion or beta-cell function (homeostasis model assessment of beta-cell function [HOMA-B], BIGTT-acute insulin response surrogate measure [BIGTT-AIR]). Changes in insulin resistance over time were not strongly related to changes in BMI or waist circumference during study drug treatment. CONCLUSIONS: The increase in insulin levels during olanzapine treatment may compensate for the increase in insulin resistance and serve to reduce fasting and postprandial glucose levels. This may contribute to the lack of differences between olanzapine and risperidone in indices of diabetic or prediabetic glucose levels or glycohemoglobin. How many years this compensatory mechanism will persist needs further investigation. Periodic OGTT tests measuring glucose and insulin levels would be helpful in assessing the status of beta-cell insulin reserve in patients treated with olanzapine and other second-generation antipsychotics and assessing an individual patient's risk for conversion to type 2 diabetes. TRIAL REGISTRATION: clinicaltrials.gov Identifier NCT00287820. PMID- 19814948 TI - Next-step strategies for panic disorder refractory to initial pharmacotherapy: a 3-phase randomized clinical trial. AB - BACKGROUND: More data are needed to guide next-step interventions for panic disorder refractory to initial intervention. METHOD: This 24-week randomized clinical trial (RCT) enrolled 46 patients with DSM-IV-defined panic disorder from November 2000 to April 2005 and consisted of 3 phases. Patients who failed to meet remission criteria were eligible for randomization in the next treatment phase. Phase 1 was a 6-week lead-in with open-label sertraline flexibly dosed to 100 mg (or escitalopram equivalent) to prospectively define treatment refractoriness (lack of remission). Phase 2 was a 6-week double-blind RCT of (1) increased-dose selective serotonin reuptake inhibitor (SSRI) versus (2) continued SSRI plus placebo. Phase 3 was a 12-week RCT of added cognitive-behavioral therapy (CBT) compared to "medication optimization" with SSRI plus clonazepam. Primary endpoints were remission and change in Panic Disorder Severity Scale (PDSS) score in the intent-to-treat sample in each phase. RESULTS: In phase 1, 20.5% (8/39) of the patients achieved remission, and only baseline severity predicted endpoint PDSS score (beta [SE] = 1.04 [0.15], t = 6.76, P < .001). In phase 2, increasing the SSRI dose did not result in greater improvement or remission rates (placebo 15% [n = 2] vs increased dose 9% [n = 1]: Fisher exact test P = NS). In phase 3, remission was minimal (medication optimization = 11% [n = 1]; CBT = 10% [n = 1]), with a lack of group difference in PDSS score reduction (t(17) = 0.51, P > .60) consistent with a small effect size (d = 0.24). CONCLUSIONS: Although power was limited and larger studies are needed, we failed to find evidence for greater benefit of increased SSRI dose versus continuation of current dose for panic disorder symptomatic after 6 weeks at moderate dose. Further, augmentation with CBT or medication optimization with clonazepam augmentation in nonremitted panic after 12 weeks of an SSRI did not differ, suggesting that both are reasonable next-step options. However, low overall remission rates in this comorbid refractory population suggest that better predictors of response to specific treatments over time and additional interventions are needed. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00118417. PMID- 19814949 TI - Easy and low-cost identification of metabolic syndrome in patients treated with second-generation antipsychotics: artificial neural network and logistic regression models. AB - OBJECTIVE: Metabolic syndrome (MetS) is an important side effect of second generation antipsychotics (SGAs). However, many SGA-treated patients with MetS remain undetected. In this study, we trained and validated artificial neural network (ANN) and multiple logistic regression models without biochemical parameters to rapidly identify MetS in patients with SGA treatment. METHOD: A total of 383 patients with a diagnosis of schizophrenia or schizoaffective disorder (DSM-IV criteria) with SGA treatment for more than 6 months were investigated to determine whether they met the MetS criteria according to the International Diabetes Federation. The data for these patients were collected between March 2005 and September 2005. The input variables of ANN and logistic regression were limited to demographic and anthropometric data only. All models were trained by randomly selecting two-thirds of the patient data and were internally validated with the remaining one-third of the data. The models were then externally validated with data from 69 patients from another hospital, collected between March 2008 and June 2008. The area under the receiver operating characteristic curve (AUC) was used to measure the performance of all models. RESULTS: Both the final ANN and logistic regression models had high accuracy (88.3% vs 83.6%), sensitivity (93.1% vs 86.2%), and specificity (86.9% vs 83.8%) to identify MetS in the internal validation set. The mean +/- SD AUC was high for both the ANN and logistic regression models (0.934 +/- 0.033 vs 0.922 +/- 0.035, P = .63). During external validation, high AUC was still obtained for both models. Waist circumference and diastolic blood pressure were the common variables that were left in the final ANN and logistic regression models. CONCLUSION: Our study developed accurate ANN and logistic regression models to detect MetS in patients with SGA treatment. The models are likely to provide a noninvasive tool for large-scale screening of MetS in this group of patients. PMID- 19814956 TI - Molecular characterisation of PFGE non-typable methicillin-resistant Staphylococcus aureus in The Netherlands, 2007. AB - In 2007 in The Netherlands, 30% of all human isolates of methicillin-resistant Staphylococcus aureus (MRSA) sent to the National Institute for Public Health and the Environment could not be typed by pulsed-field gel electrophoresis (non typable (NT)-MRSA). Molecular characterisation of the NT-MRSA isolates revealed 27 different spa types and two distinct SCCmec types, type IV and V. All NT-MRSA isolates were closely related based on spa and multi-locus sequence typing and belonged to the ST398 lineage. The rapid increase of NT-MRSA (ST398) isolates over the last years shows the importance of this relatively new clonal lineage. PMID- 19814957 TI - Trichinellosis outbreak in Lithuania, Ukmerge region, June 2009. AB - An outbreak of trichinellosis due to wild boar meat was detected in Lithuania in June 2009. The outbreak affected 107 people all of whom had consumed sausages made of wild boar meat. Inspection of food samples confirmed the presence of Trichinella larvae in the meat. PMID- 19814958 TI - Oseltamivir susceptibility in south-western France during the 2007-8 and 2008-9 influenza epidemics and the ongoing influenza pandemic 2009. AB - The recent emergence of seasonal influenza A(H1N1) strains resistant to oseltamivir makes it necessary to monitoring carefully the susceptibility of human influenza viruses to neuraminidase inhibitors. We report the prevalence of the oseltamivir resistance among influenza A viruses circulating in south-western France over the past three years: seasonal influenza A(H1N1), seasonal influenza A(H3N2), and the influenza A(H1N1)v viruses associated with the ongoing 2009 pandemic. The main result of the study is the absence of oseltamivir resistance in the pandemic H1N1 strains studied so far (n=129). PMID- 19814961 TI - ECDC in collaboration with the VAESCO consortium to develop a complementary tool for vaccine safety monitoring in Europe. PMID- 19814959 TI - Enhanced surveillance of initial cases of pandemic H1N1 2009 influenza in Ireland, April-July 2009. AB - From 28 April to 18 July 2009 there were 156 cases of pandemic H1N1 2009 influenza confirmed in Ireland. During this time, Ireland was in containment phase, and detailed case-based epidemiological information was gathered on all cases presenting in the community and acute health care setting. Active case finding was performed among contacts of cases. Eighty percent of cases were in people less than 35 years of age and 86% were imported. The most frequent symptoms were fever, sore throat, myalgia and dry cough. Nine people were hospitalized, no fatalities occurred. PMID- 19814962 TI - Laboratory support for the diagnosis and surveillance of sexually transmitted infections (STIs) in Eastern Europe. AB - This report outlines the proceedings of the 4th Annual Meeting of the Eastern European Network for Sexual and Reproductive Health (EE SRH Network) [1,2], which took place at Uppsala University in Uppsala, Sweden between 30 May and 3 June, 2009. PMID- 19814963 TI - Results of a vaccination campaign against human papillomavirus in the province of La Spezia, Liguria, Italy, March-December 2008. AB - Sexually transmitted diseases caused by human papillomavirus (HPV) are being diagnosed more frequently than others. It is accepted that HPV infection is a necessary cause for all cases of cervical carcinoma and a large number of other anogenital and oral cancers. Two vaccines have been developed and were licensed in 2007, which can prevent infections and pre-cancerous lesions due to HPV. In Italy pre-adolescent age (12 years-old) was identified as the ideal age for vaccination against HPV. In Liguria, the first free HPV vaccination campaign was started on 8 March 2008 in 12 year-old girls. We assessed the adherence to the vaccination during the 2008 campaign as 80.6%, 79.0% and 64.1%, respectively, for the first, second and third dose of vaccine in the target population. PMID- 19814964 TI - Ongoing rubella outbreak in Bosnia and Herzegovina, March-July 2009--preliminary report. AB - Between 24 March and 31 July 2009, 342 clinically diagnosed cases of rubella were notified in five municipalities in Republika Srpska, Bosnia and Herzegovina. Fourteen cases were laboratory-confirmed by positive IgG against rubella virus. Four virus isolates were obtained and identified as genotype 2B strains, with one isolate differing by a single mutation in the region of the E1 gene. This ongoing outbreak revealed gaps in the immunisation programme during the war in BiH (1992 1995) and highlights the need to revise legislation to permit immunisation of children above 14 years of age with measles, mumps, rubella (MMR) vaccine and to introduce supplemental immunisation activities. PMID- 19814965 TI - Early estimates of 2009 pandemic influenza A(H1N1) virus activity in general practice in France: incidence of influenza-like illness and age distribution of reported cases. AB - In the end of August 2009, an unusually elevated level of influenza-like illness (ILI) activity was reported to the French Sentinel Network. We quantified the observed excess in ILI cases in France during summer 2009 and characterised age patterns in reported cases. An excess of cases has been observed since 5 July, with a time increasing trend. The cumulated estimated excess number of ILI cases was 269,935 [179,585; 316,512], corresponding to 0.5% French population over the period. Compared to the same period in the past years, relative cumulated incidence was greater among young subjects and lower among subjects over 65 years old. Compared to past epidemics, the relative cumulated incidence was greater in children less than 5 years old. This excess of cases may reflect the current spread of the A(H1N1) virus in France, subject to the following limitations: estimates were based on clinical cases consulting a GP; large media coverage may have led to a non specific increase in consultation rates. PMID- 19814966 TI - Residual immunity in older people against the influenza A(H1N1)--recent experience in northern Spain. AB - The 2009 pandemic influenza A(H1N1) virus has a higher incidence in children and young adults, a pattern that has also been reported in seasonal influenza caused by the influenza A(H1N1)virus. We analysed age at infection in symptomatic patients with influenza in the Basque Country (northern Spain), reported through the sentinel influenza surveillance system which monitors 2.2-2.5%of the population. Between September 1999 and August 2009,influenza A(H3N2) or seasonal influenza A(H1N1) was detected in 941 patients, and from April to August 2009, pandemic influenza A(H1N1) was detected in 112 patients. The H3/H1 seasonal influenza ratio was between 3.3 and 3.4 in the under 60 year olds,but 9.8 in older individuals, suggesting that people born before 1950 have residual immunity against the influenza A H1N1 subtype (both seasonal and pandemic). PMID- 19814967 TI - [Aerosol therapy in treatment of childhood otitis media with effusion]. AB - INTRODUCTION AND OBJECTIVES: We address the efficacy of aerosol therapy in the treatment of otitis media with effusion during childhood. We study audiometric recovery in comparison with other classic treatments. MATERIAL AND METHODS: This is a retrospective analysis of 37 patients suffering from otitis media with effusion treated with aerosols. We analyze the pure tone audiometry gap results for the whole sample of patients. We also evaluate the characteristics of the group of patients that had previously required surgery and the group withdrawn from aerosol therapy for not responding. RESULTS: Thirty seven patients with a mean age of 6.8 years met the inclusion criteria. Audiometric tests were performed at the beginning of the treatment and after one month, 3 months and finally 6-12 months. In audiometric terms, 76% of the patients achieved results similar to those obtained after surgery. Seven patients were withdrawn from treatment due to poor or no response to aerosol therapy or due to a lack of collaboration. Two patients developed complications not related to aerosol therapy (tympanic perforation and cholesteatoma pearl). CONCLUSION: The efficacy of aerosol therapy is comparable to that obtained with classic treatments. We have found no differences in the outcomes obtained in the group previously treated with surgery. We found no indicators of poor response in those patients where the treatment failed. PMID- 19814968 TI - [Considerations on acute otitis externa for its optimized treatment]. AB - INTRODUCTION AND OBJECTIVES: To evaluate the optimal treatment for diffuse acute otitis externa (DAOE) by means of a new staging system based on clinical and otoscopic findings. METHODS: Monitoring of otitis cases diagnosed at our Centre over the last 21 months (n=1,026), taking into account those who returned for a second visit, either via the emergency service or with a prior appointment. Staging of otitis externa by degree of obstruction of the external auditory canal (EAC): under 25% (type I, n=174); between 25% and 75% (type II; n=277); over 75% (type III; n=359); total obstruction due to organized oedema (type IV; n=216). RESULTS: In most cases at an advanced stage, the canal was cleared by aspiration and placement of gauze or cellulose sponges. Ototopical drops were chosen in more than 95% of cases in every group, but anti-inflammatory and systemic antibiotic agents were administered in significantly larger amounts to patients in stages III and IV. Stage IV showed a statistically significant increase in the percentage of second visits. These manoeuvres improved pain, fullness and otorrhea significantly faster in stages I and II. CONCLUSIONS: Staging of otitis allows comparisons of the level of involvement and treatment recommendations: aspiration is advisable in stages II, III and IV; placement of a wick inside the canal is necessary in stages III and IV. Ototopical drops seem to be needed in every case and systemic antibiotics almost only in stage IV. Acidifying agents and topic glycerine vehicles access the canal better than aqueous solutions and would be the first choice in advanced stages. PMID- 19814969 TI - [Association between endolymphatic hydrops and benign paroxysmal positional vertigo: coincidence or causality?]. AB - INTRODUCTION: The aim of this study was to determine the prevalence of benign paroxysmal positional vertigo (BPPV) in patients diagnosed with endolymphatic hydrops (EH) and to analyze whether this association is a factor for poor prognosis in these patients. MATERIAL AND METHODS: A retrospective study was carried out in our department among the patients definitively diagnosed as having Meniere's disease according to the criteria of the American Academy of Otolaryngology and BPPV by the usual triggering manoeuvres. RESULTS: The prevalence of BPPV in our patients with EH was 5.1%. The rate of cure with the repositioning manoeuvres is 90.9%, however success with the first manoeuvre was only 63.36%. Recurrences occurred in up to 22.22%. We found a lower success rate with the first manoeuvre in relation to the rest of the patients with BPPV without EH. This difference is statistically significant (P=0.042). DISCUSSION: We did not find that these patients to have a lower rate of cure with the repositioning manoeuvres or a higher rate of recurrence. But in our protocol the first manoeuvre had a lower success rate than in the rest of the patients. PMID- 19814970 TI - [Inverted acoustic reflex: an analysis of its morphological characteristics in different physiological and pathological situations]. AB - INTRODUCTION AND OBJECTIVES: Inverted acoustic reflex (IAR) or inverted acoustic effect is the name given to the positive deflexion observed during the carrying out of an admittancemetry when the ear is subjected to a high intensity sound, rather than the typical negative deflexion due to the contraction of the middle ear musculature. The objective of the paper is to analyze the morphology of the IAR by relating it to the situations (physiological or pathological) in which this effect appears with the aim of establishing a pathogenic hypothesis. METHODS: We study the admittance traces obtained in 50 patients on stimulating the ear with high sound intensities (between 100 and 110 dBs HL) in different situations (ageing, death, otosclerosis, facial palsy, perception hypoacusia and normoacusia). We analyse the morphologies of the resulting traces as well as the on-set and off-set latencies of the reflexes. RESULTS: The morphology and latency parameters are similar in all cases in which IAR appears, regardless of the physical conditions of the ear but always with the prior condition of the absence of any contraction of the stapes bone muscle. The inverted acoustic effect does not disappear after deep muscular relaxation induced by anaesthesia and is present in corpses at a time when neural activity can no longer take place. CONCLUSIONS: Although some of the characteristics of IAR cannot be explained by purely mechanical hypotheses, the appearance of this effect in a corpse and its non-disappearance with muscular relaxation means that we consider its origin to be a purely mechanical effect not mediated by any muscular reflex contraction in the human middle ear. PMID- 19814971 TI - [Peripheral and central audiologic findings in patients with Vogt-Koyanagi-Harada syndrome]. AB - OBJECTIVE: To identify and describe peripheral and central audiological abnormalities of patients with Vogt Koyanagi Harada syndrome. MATERIALS AND METHODS: Prospective, cross-sectional, observational, descriptive study. Vogt Koyanagi Harada patients referred from an ophthalmologic centre were assessed for signs and symptoms of auditory pathology. Peripheral audiological and central auditory processing tests were performed. To standardize the latter, methodological controls were matched for sex, age and audiometric chart. RESULTS: 21 patients were included (3 male, 18 female), mean age 40.7 years (+/-11.82). Nine of the 21 complained of tinnitus and nine suspected hearing loss. Exclusively or mainly sensorial abnormalities of pure tone audiometric chart were documented in 61.9% (mainly selective troughs at 4 and 8 kHz); 95.3% of patients had deficit in high-frequency audiometry. Five of the patients had sensorial disorders in the speech audiometry. No central auditory processing disorder was seen. CONCLUSIONS: Most of the patients with Vogt Koyanagi Harada syndrome had objective peripheral audiological abnormalities, although few revealed any symptoms. PMID- 19814972 TI - [The role of chemotherapy in advanced oral cavity cancer]. AB - OBJECTIVE: To evaluate the role of chemotherapy in advanced cancer of the oral cavity by assessing its influence on the survival of patients receiving palliative care and its ability to improve the expectations of curative surgery. PATIENTS AND METHOD: Ninety-nine consecutive patients were analyzed to assess their course, the prognostic factors for tumour progression, and overall survival after treatment with two different chemotherapy regimes: cisplatin plus bleomycin (n=45) or cisplatin plus 5-fluorouracil (n=52). For the analysis, patients were split into groups depending on the treatment intention: palliative (n=41) or neo adjuvant (n=56). RESULTS: The response rate was 56.7%. The response was greater in the neo-adjuvant group (73.2%) than among the palliative patients (36.6%) (P<0.001); greater with the bleomycin regime (73.3%) than with 5-fluorouracil (44.2%) (P=0.003); and greater in patients with stage III (77.8%) than stage IV (50%) (P=0.021). Survival was higher among patients receiving bleomycin than in those on 5-fluorouracil (P=0.019) and among those with stage III than stage IV (P=0.013). In the palliative treatment group, the response was associated with greater survival (P<0.001). In the neo-adjuvant group, 30 patients (53.6%) underwent surgery, with complete resection in 26 of them (86.7%). Sixteen cases (61.5%) presented recurrence. Chemotherapy response was not associated with operability. Survival was higher in the group with complete tumour resection (P<0.041). CONCLUSIONS: The response to chemotherapy in oral cavity tumours was greater in patients with stage III and in those receiving bleomycin. Chemotherapy may improve survival in palliative cases. In neo-adjuvant cases, no great influence was seen in the conversion of doubtful cases into candidates for surgery. PMID- 19814973 TI - [Thyroid function alteration in patients with radiation therapy of the neck]. AB - INTRODUCTION: The aim of this study is to determine whether the incidence of hypothyroidism in patients with Head and Neck Cancer who have undergone neck radiotherapy justifies the inclusion of thyroid function monitoring in the pre operative and follow-up evaluation protocols. MATERIAL AND METHODS: A retrospective study was conducted with all the patients seen in the Oncology Section of the Otorhinolaryngology Department in the "Hospital de Clinicas", Montevideo, Uruguay, from January 1970 to May 2008. A total of 550 case records were analyzed. Of the 550 patients, 188 were excluded due to the fact they had not received radiotherapy as part of their treatment and 362 were included in the study. Of these, 55 patients had thyroid stimulating hormone (TSH) level determination after treatment with radiation therapy. Hypothyroidism was defined as a TSH value greater than or equal to 4.5 mIU/L, regardless of whether or not any symptoms were presented. RESULTS: 36.4% of the patients were diagnosed as having clinical or sub-clinical hypothyroidism. The type of treatment carried out, particularly whether or not surgical resection was performed, was found to be the most significant predictive factor for the development of hypothyroidism (P=0.054). CONCLUSION: Monitoring of the thyroid function on a six-month or annual basis, at least during the first 5 post-treatment years, has to be included in the follow-up and control protocols of every patient undergoing neck RT, and patients with TSH values over 4.5 mIU/L should be referred to the endocrinologist for hormone replacement, whether they present symptoms or not. PMID- 19814974 TI - [Prognostic significance of angiogenesis in surgically treated supraglottic squamous cell carcinomas of the larynx]. AB - INTRODUCTION AND OBJECTIVES: It is now generally accepted that angiogenesis is crucial in tumour growth. However, controversy still exists regarding the prognostic significance of angiogenesis in head and neck carcinomas. The aim of this paper is to determine the prognostic significance of angiogenesis in a homogeneously treated group of supraglottic squamous cell carcinomas. MATERIAL AND METHODS: 108 patients surgically treated for squamous cell carcinoma of the supraglottic larynx were studied. Angiogenesis was estimated in the primary tumour and in the nodal metastases by determining microvessel density using the "hot spot" method. Anti-CD34 antibody was used to stain blood vessels. RESULTS: The mean microvessel density in primary tumours was 72+/-34 vessels/mm(2) and 58.5+/-31.5 vessels/mm(2) in nodal metastases. No correlation was found between microvessel density in the primary tumours and the corresponding nodal metastasis (P=0.195). No significant differences in microvessel density were observed in relation to clinico-pathological parameters or survival (P=0.19). CONCLUSIONS: Our results suggest that microvessel density is not a useful prognostic marker in surgically treated supraglottic squamous cell carcinomas. PMID- 19814975 TI - [Baha: a new longitudinal incision with fixture out of the incision]. AB - Since the introduction of bone-anchored hearing aids (BAHA) a few decades ago, their effectiveness has made the use of these devices a well-established method. The most common complications associated with BAHA are dermatological, so a lot of changes were introduced in the surgical technique to reduce the frequency of skin reactions. We have analyzed our experience in the management of the skin and soft tissues over 8 years' experience with BAHA. We present a new longitudinal incision with the fixture out of the incision. PMID- 19814976 TI - [Reconstruction of pharyngeal defects]. AB - Reconstruction of pharyngeal defects continues to present a clinical challenge for the Head and Neck surgeon. We have different reconstructive options to preserve speech, airway and swallowing functions. Reconstructive surgery implies a balance between oncologic cure, patient morbidity and quality of life. Classical reconstructive techniques include pedicled cervical cutaneous or myocutaneous flaps and distal myocutaneous flaps such as from the pectoralis major. Current microvascular technique options have a differing incidence of complications but always with high success rates. This article reviews the most current options on reconstructive techniques in pharyngeal defects. PMID- 19814977 TI - [Tuberculous arthritis of the temporomandibular joint caused by Mycobacterium bovis. A diagnostic conundrum]. AB - Infrequent variants of extrapulmonary tuberculosis pose a challenging public health problem because of the delay in their diagnosis and treatment. Joint involvement is usually encountered in major joints (hip, elbow, knee and ankle). Tuberculous involvement of the temporomandibular joint is extremely uncommon but must be considered in patients with pain, inflammation or joint stiffness not resolved with conventional treatment. PMID- 19814978 TI - [Retropharyngeal liposarcoma: apropos of a case of apnoeas and hypopnoea during sleep]. AB - Liposarcomas rarely occur in the head and neck. We report a case of myxoid liposarcoma occurring in the retropharyngeal space in a middle-aged male with clinical symptoms of obstructive sleep apnoea. PMID- 19814979 TI - [Mucocutaneous Leishmaniasis: an imported illness with ENT repercussions]. AB - We report a case of mucocutaneous Leishmaniasis, an uncommon illness in our area. Leishmaniasis covers a group of diseases caused by protozoa of the genus Leishmania with several pathogenic species transmitted by Phlebotomus mosquitoes. Leishmania braziliensis is endemic in parts of South America and is responsible for the mucocutaneous Leishmaniasis reported here. The initial lesion is cutaneous and appears as an ulcer on arms or legs, leaving a scar. In some untreated cases, a late phase may affect the ENT area with hard to treat chronic destructive and mutilating lesions. PMID- 19814980 TI - [Solitary extramedullary plasmocytoma of tubaric tonsil]. AB - Several histopathological modalities of tumours can grow in the rhinopharynx. Solitary extramedullary plasmocytoma (SEP) is a low frequent variety of low-grade lymphoma. The aim of this work is to present a new case of SEP localized in rhinopharynx and to perform a review of diagnostic and therapeutic protocols of these neoplasias. They must be taken into account in the differential diagnosis of nasopharyngeal tumours. PMID- 19814981 TI - [Vestibular findings in patients with Vogt-Koyanagi-Harada syndrome]. AB - OBJECTIVE: To identify and describe vestibular abnormalities in patients with Vogt-Koyanagi-Harada syndrome (VKH). MATERIALS AND METHOD: Prospective, cross sectional, observational and descriptive study. Patients with VKH referred by an ophthalmological center, were interrogated and physically examined in search of signs of vestibular abnormalities, and if positive, they underwent videonystagmography, computerized dynamic posturography, tonal audiometry and tympanometry. RESULTS: Out of 21 patients with VKH, only 10 were included in the study due to presenting data of vestibular abnormalities (10/10 with vestibular symptoms and 9/10 with abnormalities in the physical exploration). The age average was 37.8 years. The videonystagmography was mainly abnormal in ocular saccades test (10/10). The posturography showed a higher alteration of the visual (4/10) and vestibular (4/10) afferents. A diagnosis of benign paroxysmal positional vertigo was mostly concluded (6/10). None presented abnormalities of the middle ear nor data of central pathology, 6/10 presented abnormalities in tonal audiometry. CONCLUSIONS: Peripheral vestibular disorder is often present in the population with VKH. PMID- 19814982 TI - [Paediatric cochlear implantation in the critical period of the auditory pathway, our experience]. AB - INTRODUCTION: Numerous experimental and clinical studies have suggested a critical or sensitive period in which the auditory pathway develops its greatest potential in terms of plasticity and learning. Early cochlear implantation performed in prelingual deaf children in this period provides a better prognosis for language acquisition. The aim of this study is to show the importance of cochlear implantation before this critical period ends. METHODS: We conducted an observational, longitudinal, retrospective study of 57 children suffering profound prelingual bilateral sensorineural hearing loss who had received Advanced Bionics implants at our ENT department between June, 1998, and November, 2006. Data on their audiometric thresholds, the disyllabic word test adapted to children, open-set sentences recognition test and the Nottingham scale were analyzed. RESULTS: The analysis of audiometric thresholds showed no differences between children receiving the implants at different ages. However, statistically significant differences (p<0.05) were found in speech tests between groups of children receiving the implants before and after 4 years of age. CONCLUSIONS: Our results are in line with other publications showing differences in auditory performance when comparing children with early implants versus children receiving the implants at a later age. We found the greatest differences at 4 years of age. Nevertheless, these findings should not exclude children over this age from implantation. PMID- 19814983 TI - [CSF fistulae following surgery for cerebellopontine angle tumours and their relationship with the body mass index]. AB - INTRODUCTION: The most frequent complication after vestibular schwannoma surgery is cerebrospinal fluid (CSF) fistula. MATERIAL AND METHODS: Retrospective study of 170 patients who had vestibular schwannoma (163) or other tumours in the cerebello pontine angle (CPA) (7). Resection was carried out using different approaches: retrosigmoid (66%), translabyrinthine (24%), middle cranial fossa (5%) and others (5%). We studied the day of onset, location and treatment of the CSF leaks, and the influence of demographic, radiological and clinical variables, including Body Mass Index. RESULTS: 27 patients developed a CSF fistula (15.9%): 15 were incisional (8.8%), 8 patients developed CSF rhinorrhea (4.7%), 1 CSF otorrhea (0.6%) and 3 a combination of the above (1.8%). We controlled 11 CSF fistulae with bed rest and compressive dressings (6.5%), 7 required lumbar drainage (4.1%), 2 lumbar drainage and wound closure with local anaesthesia (1.2%) and 7 patients required returning to the operating room under general anaesthesia (4.1%). There was not a significant relationship between the apparition of CSF and tumour size, type of approach, age or body mass index (BMI). CONCLUSIONS: Despite the great development and new surgical techniques, CSF fistulae are still a frequent complication after VS surgery. There might be other aetiological factors such as intracranial pressure. There was no significant relationship between CSF fistula and BMI. Adequate management and early conservative measures led to reduced reintervention rates of less than 5% in all patients. PMID- 19814984 TI - [Neurocognitive and behavioural abnormalities in paediatric sleep-related breathing disorders]. AB - INTRODUCTION: Behavioural and neurocognitive abnormalities in children may be a consequence of sleep-related breathing disorders. The effectiveness of assessments based on questioning parents is dubious and objective assessment tools are therefore required. AIM: To ascertain the impact of these abnormalities in children with sleep-related breathing disorders and compare the reliability of questioning parents in relation to validated psychological tests. METHOD: A prospective study was performed on 20 children with sleep-related breathing disorders and 20 healthy control children between 3 and 12 years of age. Both groups were subjected to a battery of validated psychological tests. The results of both groups were compared with each other and with the response to clinical questionnaires given to parents in the problem group. RESULTS: More than 75% of the cases in the problem group presented abnormalities with regard to attention, anxiety, memory and spatial structuring. The percentage involvement in all concepts was higher in the problem group. Comparisons of attention (40% of children affected in the control group and 80% in the problem group), memory (50% and 84.2%), and spatial structuring (45% and 75%) were statistically significant. More abnormality was observed in the parameters assessed with psychological tests than the equivalent concept obtained from interviewing the parents. Comparison of abnormal concentration assessed from the questionnaires (40% of children affected) with attention during the psychological test (80%), memory (15% and 84.21%), and delayed language development (10%) compared to spatial structuring (75%) was statistically significant. CONCLUSIONS: A high prevalence of behavioural and neurocognitive abnormalities was observed in children with sleep related breathing disorders compared to a control group of healthy children. The use of objective assessment such as psychological tests revealed more abnormalities than were expressed by parents in response to clinical interviews. PMID- 19814985 TI - [Paranasal sinuses mucoceles. Our experience in 72 patients]. AB - INTRODUCTION: Traditional treatment for paranasal sinuses mucoceles recommended total removal through external approaches. Since the 90s, endoscopic marsupialization has been proposed as optimal surgical treatment. We present our experience in the treatment of this pathology. MATERIAL AND METHOD: A retrospective review of 72 patients treated for paranasal sinuses mucoceles between 1980 and 2006 in our ENT department was performed. We describe clinical features, surgical approaches employed and recurrence of disease. RESULTS: The sample was composed of 72 patients with average follow-up period of 44 months (range 13-214 months). A total of 81 mucoceles were presented, with 44% affecting the frontal sinus or frontoethmoidal cells, followed in frequency by maxillary sinus mucoceles (35%). Twenty-nine percent of the patients did not present predisposing factors; 31% of patients had a history of nasal polyposis, 35% had undergone previous sinus surgery and 14% suffered previous facial fractures; 48 mucoceles patients were treated endoscopically and 33 were treated with external or combined approaches. Recurrence was found in 7 patients, 2 in the endoscopic surgery group and 5 in the external/combined surgery group. CONCLUSIONS: Endoscopic marsupialization is a safe approach with a low rate of recurrence. The endoscopic approach may be unsuitable for frontal lateral sinus mucoceles or those with significant bone blockage. PMID- 19814986 TI - [Surgical management of salivary gland tumours]. AB - INTRODUCTION AND OBJECTIVES: Salivary gland tumours account for 3% of head and neck tumours. The aim of this study is to analyze our series of tumours of the salivary glands requiring surgical treatment at our centre from 2004 to 2007. MATERIAL AND METHODS: This study is a retrospective review of 49 patients diagnosed as having tumours of the major and minor salivary glands or ectopic salivary tumours, and surgically treated at our hospital between 2004 and 2007. We reviewed their clinical characteristics, imaging findings, fine-needle cytology results, surgical charts (sub-maxillectomies, parotidectomies, palatal tumour excisions and cervicotomies), final pathology findings, and course with at least one year follow-up. RESULTS: Forty-nine salivary tumours were treated, including 43 parotid tumours (87%), 3 sub-mandibular tumours (6%), 1 palatal tumour (2%) and 2 ectopic tumours (4%). Sixteen percent of the tumours were malignant. Fine-needle cytology sensitivity was 40%, whereas specificity was 100%. Out of 43 parotidectomies, 40 (93%) were primary parotidectomies, and 3 were revision parotidectomies. Most parotidectomies (81%) were superficial or partial and 8 (19%) were total. Parotidectomy complications are similar to those described previously in the literature: permanent facial palsy in superficial or partial parotidectomy (5%), wound dehiscence or necrosis (13%), post-operative bleeding (4%), fever or wound infection (7%), sialoceles (44%) and Frey's syndrome (2%). CONCLUSIONS: Parotid tumours are the most common salivary gland tumours. Most of them are benign, but Warthin's tumour is more frequent than usual in our series. Early and late complications from parotidectomy are uncommon, although sialocele is a common transitory complication in our series. PMID- 19814987 TI - [Radiological diagnostic of the non-pathological conditions of the petrous apex]. AB - Many patients with otological symptoms are remitted to the otolaryngology outpatient clinics every day. These patients commonly undergo imaging studies, generally magnetic resonance imaging (MRI). In some cases, a positive unilateral result is found in the form of a potentially pathological signal that can be observed in the petrous apex region. We present the cases of 6 patients (aged between 26 and 62 years) with asymmetric bone marrow distribution or trapped mucous fluid secretions in the petrous apex, collected over a 6-year period. Diagnosis was made with the use of CT scans and MRI. All of the patients were referred for skull base surgery. In all cases a non-pathologic asymmetry was diagnosed in the petrous apex. Certain non-pathologic conditions of the petrous apex must be treated expectantly without any surgery. PMID- 19814988 TI - [Communication of needs in laryngectomized patients: Pictorrino]. AB - OBJECTIVE: To design, create and validate a pictogram, called Pictorrino, as a tool to obtain direct and simple communication with laryngectomized patients. PATIENTS AND METHODS: In a prospective sample of 10 laryngectomized patients within the first postoperative week, their principal needs and demands were evaluated, and were confirmed in a retrospective second sample of 10 laryngectomized patients who attended reviews in the outpatient clinic. Thereafter, pictograms were created to state such needs in as clear a fashion as possible. RESULTS: A pictogram was designed and validated, named and registered as Pictorrino, consisting of a board, which showed these pictograms on one of its sides, and a visual analogue scale of pain, with the aim of enabling the patient to express the demand or need at every moment. CONCLUSIONS: With Pictorrino we have achieved a multicultural tool that allows a more direct communication with laryngectomized patients. PMID- 19814989 TI - [Optical coherence tomography applications in otolaryngology]. AB - Optical coherence tomography is a novel imaging technique providing high resolution bidimensional images of tissue microstructures. Several studies have been published on the use of this technique in different fields of medicine, particularly ophthalmology. There are very few studies in the field of otolaryngology. This paper presents various applications of optical coherence tomography in the different sub-specialties of otolaryngology, as well as the benefits of this technique over traditional diagnostic methods. PMID- 19814990 TI - [Analysis of the otorhinolaryngology services authorized for the training of residents]. AB - The aim of this study was to obtain information on the current educational offer of the authorized Units with the intention of evaluating their teaching capacity and identify their weaknesses and shortcomings. For this purpose an electronic, self-completing questionnaire was sent to the various teaching units. In addition, information on the most important aspects of the management of hospitals was also collected. Fifty-eight forms were received and except for 5 cases the information from the management of the hospitals was also received. The resources for external consultation, the number of special examinations in Audiology, Speech and Otoneurology, the resources in the operating room and the number of surgical interventions as well as the scientific activity developed in the last 5 years, was outlined. From the figures obtained, some critical areas were identified for the training of current residents in otolaryngology, which were also scored. Considering a threshold of 5 points, excluding the performance of several of these basic requirements, 19 services were below the threshold. PMID- 19814991 TI - [Tonsillar actinomycosis manifested as expectorated debris]. AB - Actinomycosis is a festering bacterial infection frequently affecting the cervicofacial area, for which the germs responsible are Gram-positive bacilli of Actinomyces sp. We present a case of atypical presentation of actinomycosis, in the shape of repetitive mass in the tonsillar fossa with complex therapeutic management. PMID- 19814992 TI - [Abscess of cavum and ophthalmoplegia secondary to malignant external otitis]. AB - We present a rare case of ophthalmoplegia, labyrinthitis and abscess of cavum secondary to skull base osteomyelitis by malignant external otitis. Since symptoms persisted in spite of antibiotic therapy, surgical drainage using a transnasal endoscopic approach was performed. PMID- 19814993 TI - [Parapharyngeal extension of a giant tracheocele]. PMID- 19814994 TI - [Tympanic blast injury]. PMID- 19814995 TI - [Consolidate and open new pathways]. PMID- 19814996 TI - Substrate discrimination by ergothioneine transporter SLC22A4 and carnitine transporter SLC22A5: gain-of-function by interchange of selected amino acids. AB - ETT (originally designated as OCTN1; human gene symbol SLC22A4) and CTT (OCTN2; SLC22A5) are highly specific transporters of ergothioneine and carnitine, respectively. Despite a high degree of sequence homology, both carriers discriminate precisely between substrates: ETT does not transport carnitine, and CTT does not transport ergothioneine. Our aim was to turn ETT into a transporter for carnitine and CTT into a transporter for ergothioneine by a limited number of point mutations. From a multiple alignment of several mammalian amino acid sequences, those positions were selected for conversion that were momentously different between ETT and CTT from human but conserved among all orthologues. Mutants were expressed in 293 cells and assayed for transport of ergothioneine and carnitine. Several ETT mutants clearly catalyzed transport of carnitine, up to 35% relative to wild-type CTT. Amazingly, complementary substitutions in CTT did not provoke transport activity for ergothioneine. In similar contrast, carnitine transport by CTT mutants was abolished by very few substitutions, whereas ergothioneine transport by ETT mutants was maintained even with the construct most active in carnitine transport. To explain these results, we propose that ETT and CTT use dissimilar pathways for conformational change, in addition to incongruent substrate binding sites. In other words, carnitine is excluded from ETT by binding, and ergothioneine is excluded from CTT by turnover movement. Our data indicate amino acids critical for substrate discrimination not only in transmembrane segments 5, 7, 8, and 10, but also in segments 9 and 12 which were hitherto considered as unimportant. PMID- 19814997 TI - Heteronemin, a spongean sesterterpene, inhibits TNF alpha-induced NF-kappa B activation through proteasome inhibition and induces apoptotic cell death. AB - In this study, we investigated the biological effects of heteronemin, a marine sesterterpene isolated from the sponge Hyrtios sp. on chronic myelogenous leukemia cells. To gain further insight into the molecular mechanisms triggered by this compound, we initially performed DNA microarray profiling and determined which genes respond to heteronemin stimulation in TNFalpha-treated cells and which genes display an interaction effect between heteronemin and TNFalpha. Within the differentially regulated genes, we found that heteronemin was affecting cellular processes including cell cycle, apoptosis, mitogen-activated protein kinases (MAPKs) pathway and the nuclear factor kappaB (NF-kappaB) signaling cascade. We confirmed in silico experiments regarding NF-kappaB inhibition by reporter gene analysis, electrophoretic mobility shift analysis and I-kappaB degradation. In order to assess the underlying molecular mechanisms, we determined that heteronemin inhibits both trypsin and chymotrypsin-like proteasome activity at an IC(50) of 0.4 microM. Concomitant to the inhibition of the NF-kappaB pathway, we also observed a reduction in cellular viability. Heteronemin induces apoptosis as shown by annexin V-FITC/propidium iodide staining, nuclear morphology analysis, pro-caspase-3, -8 and -9 and poly(ADP ribose) polymerase (PARP) cleavage as well as truncation of Bid. Altogether, results show that this compound has potential as anti-inflammatory and anti cancer agent. PMID- 19814998 TI - Measuring levels of proteins by various technologies: can we learn more by measuring turnover? AB - In routine experiments, scientists measure the levels of various substances such as proteins after various treatments. Detection of a change in levels suggests an impact of treatment on that particular protein. However, we sometimes forget the importance of turnover in this process. Proteins have half-lives that may change in response to treatments (which is in fact why levels may change), and an examination of half-lives may yield better clues as to how treatment affects the protein. After an exploration of the quantitative aspects of protein turnover, several interesting conclusions may be drawn. (1) Even though levels of some proteins may NOT change after treatments, their half-lives and turnovers do change, and these may be more sensitive indicators of the impact of treatments on the proteins of interest. (2) Treatments can affect protein levels because they alter either the synthesis or degradation of the protein or both. But, the rate of change of the levels depends on the half-life of the protein. If the experimenter waits only a fraction of a half-life of the protein after treatment, no significant change in level may be found since it can take up to 5 half-lives for the protein level to adjust to about 97% of its new level after treatment. (3) Half-lives of the same protein can vary in different species and experimental conditions may have to be altered if using different species. These factors suggest that a consideration of protein turnover and half-lives will be useful for future studies of this type. PMID- 19814999 TI - Chain termination and inhibition of mammalian poly(A) polymerase by modified ATP analogues. AB - We report the inhibition of mammalian polyadenylation by the triphosphate derivatives of adenosine analogues, 8-chloroadenosine (8-Cl-Ado) and 8 aminoadenosine (8-amino-Ado), which are under preclinical and clinical investigations for the treatment of hematological malignancies. The nucleotide substrate specificity of bovine poly(A) polymerase (PAP) towards C8-modified ATP analogues was examined using primer extension assays. Radiolabeled RNA primers were incubated with bovine PAP, and in the absence of ATP, no primer extension was observed with 8-Cl-ATP, whereas 8-amino-ATP resulted in chain termination. The effects of modified ATP analogues on ATP-dependent poly(A)-tail synthesis by bovine PAP also were determined, and incubation with analogue triphosphate resulted in significant reduction of poly(A)-tail length. To model the biochemical consequences of 8-Cl-Ado incorporation into RNA, a synthetic RNA primer containing a 3'-terminal 8-Cl-AMP residue was evaluated, and polyadenylation of the primer by bovine PAP with ATP was blocked completely. To explain these experimental observations and probe the possible structural mechanisms, molecular modeling was employed to examine the interactions between PAP and various ATP analogues. Molecular docking demonstrated that C8 modifications of ATP led to increased distance between the 3'-hydroxyl group of the RNA oligonucleotide terminus and the alpha-phosphate of ATP that render the molecules in an unfavorable position for incorporation into RNA. Similarly, C8 substitution with a chlorine or amino group at the 3'-terminal residue of RNA also inhibits further chain elongation by PAP. In conclusion, modified ATP analogues may exert their biological effects through polyadenylation inhibition, and thus may provide an RNA-directed mechanism of action for 8-Cl-Ado and 8-amino Ado. PMID- 19815000 TI - Magnolol and honokiol prevent learning and memory impairment and cholinergic deficit in SAMP8 mice. AB - The therapeutic use of neurotrophic factors to treat neurodegenerative disorders, including Alzheimer's disease, is considered feasible. Magnolol and honokiol, constituents of the Magnolia plant, are small organic compounds with neurotrophic activity. We investigated whether magnolol and honokiol can prevent age-related learning and memory impairment and cholinergic deficits in senescence-accelerated mice (SAM). Magnolol (1, 10 mg/kg) or honokiol (0.1, 1 mg/kg) were orally administered to SAMP8 mice once a day for 14 days in 2-month-old mice. Learning and memory performance were evaluated by passive avoidance tests and location and object novelty recognition tests. SAMP8 mice showed significant impairment of learning and memory at 4 and 6 months of age. This age-related learning and memory impairment was prevented by pretreatment with either magnolol (10 mg/kg) or honokiol (1 mg/kg). Cholinergic neuron densities in the medial septum and vertical limb of the diagonal band of the forebrain were evaluated by an immunohistochemical analysis of choline acetyltransferase (ChAT). SAMP8 mice showed a significant cholinergic deficit at 6 months of age. These age-related cholinergic deficits were prevented by treatment with either magnolol (10 mg/kg) or honokiol (1 mg/kg). Moreover, SAMP8 mice showed decreased activity of Akt, a member of the prosurvival pathway, in the forebrain at 2 months of age. A 14-day treatment with either magnolol (10 mg/kg) or honokiol (1 mg/kg) enhanced phosphorylation of Akt in the forebrain at 2 months of age. These results suggest that magnolol and honokiol prevent age-related learning and memory impairment by preserving cholinergic neurons in the forebrain. These compounds may have potential therapeutic applications to various neurodegenerative disorders. PMID- 19815002 TI - Tar DNA binding protein of 43 kDa (TDP-43), 14-3-3 proteins and copper/zinc superoxide dismutase (SOD1) interact to modulate NFL mRNA stability. Implications for altered RNA processing in amyotrophic lateral sclerosis (ALS). AB - Amyotrophic lateral sclerosis (ALS) is a fatal neurological disease characterized by progressive motor neuron degeneration in association with neurofilament (NF) aggregate formation. This process is accompanied by an alteration in the stoichiometry of NF subunit protein expression such that the steady state levels of the low molecular weight NF (NFL) mRNA levels are selectively suppressed. We have previously shown that each of TDP-43, 14-3-3 and mutant SOD1 can function as NFL mRNA 3'UTR binding proteins that directly affect the stability of NFL transcripts. In this study, we demonstrate that the interaction of TDP-43 with the NFL mRNA 3' UTR involves ribonucleotide (UG) motifs present on stem loops of the 3'UTR as well as the RRM1 and RRM2 motifs of TDP-43. Ex vivo, TDP-43, 14-3-3 and SOD1 proteins interact to modulate NFL mRNA stability, although in vivo, only TDP-43 and either mutant or wild-type SOD1 co-localize in ALS motor neurons. TDP 43 was observed to co-localize to RNA transport granules (Staufen immunoreactive) in both control and ALS spinal motor neurons. In contrast, both stress granules (TIA-1 immunoreactive) and processing bodies (P-bodies; XRN-1 immunoreactive) were more prevalent in ALS motor neurons than in controls and demonstrated strong co-localization with TDP-43. Using RNA-IP-PCR, we further demonstrate that NFL mRNA is preferentially sequestered to both stress granules and P-bodies in ALS. These data suggest that NFL mRNA processing is fundamentally altered in ALS spinal motor neurons to favour compartmentalization within both stress granules and P-bodies, and that TDP-43 plays a fundamental role in this process. PMID- 19815003 TI - Aromatase expression in the normal and epileptic human hippocampus. AB - Aromatase is a key enzyme in estrogen biosynthesis that is involved in neuronal plasticity in the rodent hippocampus. Although aromatase mRNA expression has been detected in the human hippocampus, its cellular distribution has yet to be determined. Here, we have examined the immunohistochemical distribution of aromatase in the normal and the epileptic and sclerotic human hippocampus. In both the normal and epileptic hippocampus, aromatase was detected in numerous CA1 CA3 pyramidal neurons, in granule cells of the dentate gyrus and in interneurons that co-expressed the calcium-binding proteins calbindin, calretinin or parvalbumin. However, only a small subpopulation of astrocytes was immunoreactive for aromatase in either the normal and epileptic hippocampus. The widespread expression of aromatase in a large population of neurons in the normal and damaged hippocampus suggests that local estrogen formation may play an important role in human hippocampal function. PMID- 19815001 TI - Lateral hypothalamic orexin/hypocretin neurons: A role in reward-seeking and addiction. AB - Orexins (synonymous with hypocretins) are recently discovered neuropeptides made exclusively in hypothalamus. Behavioral, anatomical, and neurophysiological studies show that a subset of these cells, specifically those in lateral hypothalamus (LH), are involved in reward processing and addictive behaviors. Fos expression in LH orexin neurons varied in proportion to conditioned place preference (CPP) for morphine, cocaine, or food. This relationship occurred both in drug-naive rats and in animals during protracted morphine withdrawal, when drug preference was elevated but food preference was decreased. Inputs to the LH orexin cell field from lateral septum and bed nucleus of the stria terminalis were Fos-activated during cocaine CPP in proportion to the preference expressed in each animal. This implies that these inputs may be involved in driving the conditioned responses in LH orexin neurons. Related studies showed that LH orexin neurons that project to ventral tegmental area (VTA) had greater Fos induction in association with elevated morphine preference during protracted withdrawal than non-VTA-projecting orexin neurons, indicating that the VTA is an important site of action for orexin's role in reward processing. In addition, stimulation of LH orexin neurons, or microinjection of orexin into VTA, reinstated an extinguished morphine preference. In self-administration studies, the orexin 1 receptor antagonist SB-334867 (SB) blocked cocaine-seeking induced by discrete or contextual cues previously associated with cocaine, but not by a priming injection of cocaine. There was no effect of SB on cocaine self-administration itself, indicating that it did not interfere with the drug's reinforcing properties. Neurophysiological studies revealed that locally applied orexin often augmented responses of VTA dopamine (DA) neurons to activation of the medial prefrontal cortex (mPFC), consistent with the view that orexin facilitates activation of VTA DA neurons by stimulus-reward associations. This LH-to-VTA orexin pathway was found to be necessary for learning a morphine place preference. These findings are consistent with results showing that orexin facilitates glutamate-mediated responses, and is necessary for glutamate dependent long-term potentiation in VTA DA neurons. We surmise from these studies that LH orexin neurons play an important role in reward processing and addiction and that LH orexin cells are an important input to VTA for behavioral effects associated with reward-paired stimuli. PMID- 19815004 TI - Overexpression of hippocampal cholinergic neurostimulating peptide in heterozygous transgenic mice increases the amount of ChAT in the medial septal nucleus. AB - Acetylcholine modulates neural activity in the hippocampal glutamatergic pathway via the induction of phosphorylated Erk and may act as a novel transmitter in septohippocampal memory formation. However, how acetylcholine synthesis in the septal nucleus is regulated is unknown. We have purified a peptide from the hippocampus of the young adult rat, named hippocampal cholinergic neurostimulating peptide (HCNP) that induces acetylcholine synthesis in vitro in the septal nucleus. Previously, levels of this peptide and/or precursor protein were reported to be decreased, and the protein to be nitrated in the brains of patients with Alzheimer's disease. Here, to clarify the involvement in the regulation of acetylcholine synthesis in vivo in the medial septal nucleus, we generated HCNP precursor transgenic mice, using a Ca2+ calmodulin-dependent protein kinase II genomic promoter. The amount of cholineacetyltransferase (ChAT) in the medial septal nucleus was increased in heterozygous HCNP transgenic mice, compared with non-transgenic littermates. This result suggests that HCNP is involved in regulating acetylcholine synthesis in vivo in the medial septal nucleus and, as such, is important for memory function. PMID- 19815005 TI - Effects of epigallocatechin gallate on tissue protection and functional recovery after contusive spinal cord injury in rats. AB - Recent studies revealed the neuroprotective effects of epigallocatechin gallate (EGCG) on a variety of neural injury .The purpose of this study was to determine the effects of EGCG on the tissue protection and behavioral improvement after spinal cord injury (SCI). Rats were randomly divided into four groups of 18 rats each as follows: sham-operated group, trauma group, and EGCG treatment groups (50 mg/kg, i.p., immediately and 1 hour after SCI). Spinal cord samples were taken 24 hours after injury and studied for determination of malodialdehyde (MDA) levels, immunohistochemistry of Bax and Bcl-2, and TUNEL reaction. Behavioral testing was performed weekly up to 6 weeks post-injury. Then, the rats were euthanized for histopathological assessment. The results showed that MDA levels were significantly decreased in EGCG treatment groups. Greater Bcl-2 and attenuated Bax expression could be detected in the EGCG-treated rats. EGCG significantly reduced TUNEL-positive rate. Also, EGCG significantly reduced the percentage of lesion area and improved behavioral function than the trauma group. On the basis of these findings, we propose that EGCG may be effective in protecting rat spinal cord from secondary injury. PMID- 19815006 TI - Remarkable increase in 3-nitrotyrosine in the cerebrospinal fluid in patients with lacunar stroke. AB - The goal of our study was whether free radicals contribute to the pathogenesis of the lacunar stroke to investigate the day after hospitalization, the concentrations of 3-nitrotyrosine and tyrosine in the cerebrospinal fluid (CSF) from living patients. The subjects included 20 living patients with lacunar stroke and 20 controls. The NIH stroke scale score was used to assess the severity of the stroke, including that the patients were mild cases. There was no expansion of the infarct lesion in the brain, as assessed by CT on the day following admission. The concentration of 3-nitrotyrosine was significantly higher in patients with lacunar stroke. In contrast, the concentration of tyrosine did not differ between the two groups. Furthermore, the 3 nitrotyrosine/tyrosine ratio was significantly higher in patients with lacunar stroke than in controls. Our results show that free radicals are produced in the CSF of lacunar stroke patients and that nitration of neuronal proteines is enhanced under this condition. These obsetvations suggest that lacunar stroke patients should be treated with edaravon, which is a free radical scavenger usually prescribed for cases of major strokes, as it will likely improve the prognosis of these patients. PMID- 19815007 TI - Sex differences in NADPH-diaphorase activity in the rat posterodorsal medial amygdala. AB - The rat posterodorsal medial amygdala (MePD) is a sexually dimorphic area implicated in the control of reproduction. Interestingly, nitric oxide (NO) synthetizing neurons are widely distributed in brain regions involved with the modulation of sexual behavior. Here we studied the NADPH-diaphorase (NADPH-d) activity and the number of positive cells in the MePD of adult males and adult females either across the estrous cycle (diestrus, proestrus, estrus, and metaestrus) or following ovariectomy and substitutive therapy (consisting of oil, estradiol alone, the combination of estradiol and progesterone, or progesterone alone). The NADPH-d histochemical technique was followed by a semi-quantitative analysis using optical densitometry. Males showed a higher MePD regional optical density and neuronal optical density than females across the estrous cycle, with the exception of the diestrus phase (P<0.01). No differences were found in these parameters during the ovarian cycle (P>0.05). There were no statistically significant differences among males and cycling females in the number of NADPH-d positive cells (P>0.05). Additionally, no statistically significant difference was found in the regional optical density, in the neuronal optical density, or in the number of NADPH-d positive neurons when comparing the data from ovariectomized females that received vehicle or the three different hormonal replacement therapies (P=0.07, P=0.18, and P=0.95, respectively). Results suggest that NADPH-d activity in the rat MePD is different between sexes but in females it is not affected by changing levels of circulating gonadal hormones in physiological or supraphysiological conditions. PMID- 19815008 TI - Dopaminergic neurotoxicity of HIV-1 gp120: reactive oxygen species as signaling intermediates. AB - We examined the role of reactive oxygen species (ROS) in loss of dopaminergic neurons (DNs) from the substantia nigra (SN) in neuroAIDS. The frequency of Parkinson-like symptomatology, and DN loss, in neuroAIDS is often attributed to nonspecific DN fragility to oxidative stress. Cultured DN are more sensitive to ROS than non-dopaminergic neurons (RN): DN underwent apoptosis at far lower H(2)O(2) concentrations than RN. Gene delivery of glutathione peroxidase (GPx1), which detoxifies H(2)O(2), largely protected both neuron types. HIV-1 envelope, gp120, which elicits oxidative stress in neurons, caused apoptosis more readily in DN than in RN. However, unlike apoptosis caused by H(2)O(2), gp120-induced DN apoptosis was specific: DNs were specifically more sensitive than RN to receptor mediated [Ca(2+)](i) fluxes triggered by gp120. Gp120-induced Ca(2+) signaling in both neuron types was inhibited by GPx1 or Cu/Zn superoxide dismutase (SOD1), implicating superoxide and peroxide in ligand (gp120)-induced signaling upstream of Ca(2+) release from intracellular stores. In vivo, rats given 10 ng of gp120 stereotaxically showed rapid DN loss within the SN, while loss of RN in the SN and caudate-putamen (CP) was slower and required > or =100 ng of gp120. Furthermore, gp120 injected into the CP was transported axonally retrograde to the SN, causing delayed DN loss there. This, too, was prevented by SOD1 or GPx1. DNs are therefore specifically hypersensitive to gp120-induced apoptosis, signaling for which involves ROS intermediates. These findings may help explain why DN loss and Parkinson's-like dysfunction predominate in neuroAIDS and may apply to other neurodegenerative diseases involving the SN. PMID- 19815009 TI - Glutamate, excitotoxicity, and programmed cell death in Parkinson disease. PMID- 19815010 TI - A bilateral cervical contusion injury model in mice: assessment of gripping strength as a measure of forelimb motor function. AB - Here, we describe a bilateral cervical contusion model for mice. Adult female mice received graded bilateral contusion injuries at cervical level 5 (C5) using a commercially available impactor (the IH device). Three separate experiments were carried out to define conditions that produce impairments in forelimb function without unacceptable impairment of general health. A grip strength meter (GSM) was used to assess gripping ability as a measure of forelimb motor function; lesion size was assessed histologically by staining cross sections for H&E and GFAP. In Experiment 1, mice received injuries of 30 kilodynes (kdyn); these produced minimal deficits on grip strength. In Experiment 2, mice received injuries of 75 kdyn and 100 kdyn. Injuries of 75 kdyn produced transient deficits in gripping that recovered between 3 and 15 days post-injury (dpi) to about 90% of control; injuries of 100 kdyn produced deficits that recovered to about 50% of control. In Experiment 3, none of the mice that received injuries of 100 kdyn recovered gripping ability. Histological assessment revealed graded injuries that ranged from damage limited primarily to the dorsal column (DC) to damage to the DC, grey matter, ventral column and lateral column. Most lesions filled in with a fibrous tissue matrix, but fluid-filled cystic cavities were found in 13% of the 100 kdyn injury group and a combination of fibrous-filled/fluid-filled cystic cavities were found in 22% and 38% of the 75-kdyn and 100-kdyn injury groups, respectively. There was minimal urine retention following cervical contusion injuries indicating preservation of bladder function. Our results define conditions to produce graded bilateral cervical contusion injuries in mice and demonstrate the usefulness of the GSM for assessing forelimb motor function after cervical contusions. PMID- 19815011 TI - Influence of the vanilloid receptor TRPV1 on the activation of spinal cord glia in mouse models of pain. AB - Although activation of spinal glia has been implicated in the development of pathological pain, the mechanisms underlying glial activation are not fully understood. One such mechanism may be triggered by reaction to neuroactive substances released from central axons of sensory afferents. The vanilloid receptor TRPV1, a nonselective cation channel in nociceptive sensory afferents, mediates the release of neurotransmitters, such as glutamate and CGRP in the dorsal horn, which can subsequently activate glia. To test the hypothesis that activation of spinal glia is mediated, at least in part, by TRPV1, we studied the expression of markers for microglia (ionized calcium-binding adapter molecule 1, Iba1) and astrocytes (glial fibrillary acidic protein, GFAP) in the spinal cord of TRPV1 knockout mice (KO) vs. wild-type mice (WT) in models of acute (intraplantar capsaicin), inflammatory (adjuvant-induced arthritis, AIA), and neuropathic pain (partial sciatic nerve ligation, PSNL). We found that (i) naive KO mice had denser immunostaining for both Iba1 and GFAP than naive WT mice; (ii) the immunostaining for Iba1 increased significantly in treated mice, compared to naive mice, 3 days after capsaicin and 7-14 days after AIA or PSNL, and was significantly greater in WT than in KO mice 3 days after capsaicin, 7-14 days after AIA, and 7 days after PSNL; and iii) the immunostaining for GFAP increased significantly in treated mice, compared to naive mice, 3 days after capsaicin and 14-21 days after AIA or PSNL, and was significantly greater in WT than in KO mice 14 days after AIA or PSNL. Our results suggest that TRPV1 plays a role in the activation of spinal glia in mice with nociceptive, inflammatory, and neuropathic pain. PMID- 19815012 TI - The effects of parkin suppression on the behaviour, amyloid processing, and cell survival in APP mutant transgenic mice. AB - Parkin suppression induces accumulation of beta-amyloid in mutant tau mice. We studied the effect of parkin suppression on behaviour and brain pathology in APP(swe) mutant mice. We produced double mutant mice with human mutated APP(swe)+partial (hemizygote) or total (homozygote) deletion of Park-2 gene. We studied the development, behaviour, brain histology, and biochemistry of 12- and 16-month-old animals in 6 groups of mice, with identical genetic background: wild type (WT), APP(swe) overexpressing (APP), hemizygote and homozygote deletion of Park-2 (PK(+/-) and PK(-/-), respectively), and double mutants (APP/PK(+/-) and APP/PK(-/-)). APP mice have reduced weight gain, decreased motor activity, and reduced number of entrances and of arm alternation in the Y-maze, abnormalities which were partially or completely normalized in APP/PK(+/-) and APP/PK(-/-) mice. The double mutants had similar number of mutant human APP transgene copies than the APP and levels of 40 and 80 kDa proteins; but both of them, APP/PK(+/-) and APP/PK(-/-) mice, had less plaques in cortex and hippocampus than the APP mice. APP mutant mice had increased apoptosis, proapoptotic Bax/Bcl2 ratios, and gliosis, but these death-promoting factors were normalized in APP/PK(+/-) and APP/PK(-/-) mice. APP mutant mice had an increased number of tau immunoreactive neuritic plaques in the cerebral cortex as well as increased levels of total and phosphorylated tau protein, and these changes were partially normalized in APP/PK(+/-) heterozygotic and homozygotic APP/PK(-/-) mice. Compensatory protein degrading systems such as HSP70, CHIP, and macroautophagy were increased in APP/PK(+/-) and APP/PK(-/-). Furthermore, the chymotrypsin- and trypsin-like proteasome activities, decreased in APP mice in comparison with WT, were normalized in the APP/PK(-/-) mice. We proposed that partial and total suppression of parkin triggers compensatory mechanisms, such as chaperone overexpression and increased autophagy, which improved the behavioural and cellular phenotype of APP(swe) mice. PMID- 19815013 TI - Subtle neurological and metabolic abnormalities in an Opa1 mouse model of autosomal dominant optic atrophy. AB - The ubiquitously expressed gene OPA1 is the main disease causing gene for autosomal dominant optic atrophy (ADOA). These patients present with bilateral reduction in visual acuity, central visual field defects and impaired color vision, secondary to the progressive loss of retinal ganglion cells (RGCs) and subsequent degeneration of the optic nerve. Up to now, it is not clear why a mutation in a ubiquitously expressed gene affects only RGCs and the optic nerve. Twenty-two-month-old Opa1 animals underwent a full examination following the Shirpa protocol. Weight, food intake and life span were monitored. Rotarod treadmill experiments were performed to assess neuromuscular function. Limb skeletal muscle was evaluated morphologically, mitochondrial cytochrome c oxidase (COX) activity was studied histochemically and mtDNA integrity was determined by long-range PCR. The Shirpa test showed that 33% of the Opa1 mice suffered from tremor and 52% of the Opa1 animals showed an abnormal clutching reflex. Control animals performed well in the accelerating Rotarod treadmill experiment whereas the Opa1 mice performed significantly worse. Skeletal muscle fibers were morphologically normal, had normal COX activity and showed no evidence of secondary mtDNA damage in contrast to patients with syndromic ADOA. We also found a highly significant difference in body weight. Our results demonstrate that OPA1 mutations affect not only RGCs but also other tissues and cell types, though to a lesser extent. In particular we found deficits in both neuromuscular and metabolic function. We therefore want to encourage clinicians to be vigilant about to extra-ocular manifestations in ADOA patients. PMID- 19815014 TI - Cybrid models of Parkinson's disease show variable mitochondrial biogenesis and genotype-respiration relationships. AB - Sporadic Parkinson's disease (sPD) is a nervous system-wide disease that presents with a bradykinetic movement disorder and frequently progresses to include depression and cognitive impairment. Cybrid models of sPD are based on expression of sPD platelet mitochondrial DNA (mtDNA) in neural cells and demonstrate some similarities to sPD brains. In sPD and CTL cybrids we characterized aspects of mitochondrial biogenesis, mtDNA genomics, composition of the respirasome and the relationships among isolated mitochondrial and intact cell respiration. Cybrid mtDNA levels varied and correlated with expression of PGC-1 alpha, a transcriptional co-activator regulator of mitochondrial biogenesis. Levels of mtDNA heteroplasmic mutations were asymmetrically distributed across the mitochondrial genome; numbers of heteroplasmies were more evenly distributed. Neither levels nor numbers of heteroplasmies distinguished sPD from CTL. sPD cybrid mitochondrial ETC subunit protein levels were not altered. Isolated mitochondrial complex I respiration rates showed limited correlation with whole cell complex I respiration rates in both sPD and CTL cybrids. Intact cell respiration during the normoxic-anoxic transition yielded K(m) values for oxygen that directly related to respiration rates in CTL but not in sPD cell lines. Both sPD and CTL cybrid cells are substantially heterogeneous in mitochondrial genomic and physiologic properties. Our results suggest that mtDNA depletion may occur in sPD neurons and could reflect impairment of mitochondrial biogenesis. Cybrids remain a valuable model for some aspects of sPD but their heterogeneity mitigates against a simple designation of sPD phenotype in this cell model. PMID- 19815016 TI - Acanthamoeba polyphaga is a possible host for Vibrio cholerae in aquatic environments. AB - Acanthamoeba is a genus of free-living amoebae found to be able to host many bacterial species living in the environment. Acanthamoebae and Vibrio cholerae are found in the aquatic environments of cholera endemic areas. Previously it has been shown that V. cholerae O1 and O139 can survive and grow in Acanthamoeba castellanii. The aim of this study was to examine the ability of Acanthamoeba polyphaga to host V. cholerae O1 and O139. The interaction between A. polyphaga and V. cholerae strains was studied by means of viable amoeba cell counts and viable count of the bacteria in the absence and presence of amoebae. The viable count of intracellularly growing bacteria was estimated by utilizing gentamicin assay. Electron microscopy was used to determine the localization of V. cholerae inside A. polyphaga. The results showed that A. polyphaga enhanced growth and survival of V. cholerae, which grew and survived inside the amoeba cells for 2weeks. The electron microscopy showed that A. polyphaga hosted intracellular V. cholerae localized in the vacuoles of amoeba cell. Neither the presence of V. cholerae together with A. polyphaga nor the intracellular localization of the bacteria inhibited growth and survival of A. polyphaga. The outcome of the interaction between these microorganisms may support strongly the role of A. polyphaga as host for V. cholerae O1 and O139. PMID- 19815017 TI - Deceptively simple but simply deceptive--Caenorhabditis elegans lifespan studies: considerations for aging and antioxidant effects. AB - The nematode worm Caenorhabditis elegans (C. elegans) is increasingly popular as a model organism for aging studies as well as for testing antioxidants and other compounds for effects on longevity. However, results in the literature are sometimes confusing and contradictory. This review introduces C. elegans as a model organism, discusses aspects that make it attractive for aging and antioxidant research, and addresses some problems and potential artifacts. PMID- 19815015 TI - Biological markers of amyloid beta-related mechanisms in Alzheimer's disease. AB - Recent research progress has given detailed knowledge on the molecular pathogenesis of Alzheimer's disease (AD), which has been translated into an intense, ongoing development of disease-modifying treatments. Most new drug candidates are targeted on inhibiting amyloid beta (Abeta) production and aggregation. In drug development, it is important to co-develop biomarkers for Abeta-related mechanisms to enable early diagnosis and patient stratification in clinical trials, and to serve as tools to identify and monitor the biochemical effect of the drug directly in patients. Biomarkers are also requested by regulatory authorities to serve as safety measurements. Molecular aberrations in the AD brain are reflected in the cerebrospinal fluid (CSF). Core CSF biomarkers include Abeta isoforms (Abeta40/Abeta42), soluble APP isoforms, Abeta oligomers and beta-site APP-cleaving enzyme 1 (BACE1). This article reviews recent research advances on core candidate CSF and plasma Abeta-related biomarkers, and gives a conceptual review on how to implement biomarkers in clinical trials in AD. PMID- 19815018 TI - A DNA-assisted binding assay for weak protein-protein interactions. AB - We describe a new method used for quantitating weak interactions between proteins in which the weak interaction is "assisted" by a known DNA-DNA interaction. Oligonucleotides, which are conjugated to proteins of interest, contain short complementary DNA sequences that provide additional binding energy for protein protein interactions. A stretch of unpaired bases links the protein to the hybridizing DNA sequence to allow formation of both protein-protein and DNA-DNA interactions with minimal structural interference. We validated the DNA-assisted binding method using heterodimerizing coiled-coil proteins. The method was then used to measure the predicted weak interaction between two domains of the Escherichia coli L-arabinose operon regulatory protein AraC. The interaction between domains has the expected magnitude (K(d)=0.37 mM) in the absence of arabinose. Upon addition of arabinose, we detected a weaker and unexpected interaction, which may necessitate modification of the proposed mechanism of AraC. The DNA-assisted binding method may also prove useful in the study of other weak protein-protein interactions. PMID- 19815019 TI - Structure and function of the oxidoreductase DsbA1 from Neisseria meningitidis. AB - Neisseria meningitidis encodes three DsbA oxidoreductases (NmDsbA1-NmDsbA3) that are vital for the oxidative folding of many membrane and secreted proteins, and these three enzymes are considered to exhibit different substrate specificities. This has led to the suggestion that each N. meningitidis DsbA (NmDsbA) may play a specialized role in different stages of pathogenesis; however, the molecular and structural bases of the different roles of NmDsbAs are unclear. With the aim of determining the molecular basis for substrate specificity and how this correlates to pathogenesis, we undertook a biochemical and structural characterization of the three NmDsbAs. We report the 2.0-A-resolution crystal structure of the oxidized form of NmDsbA1, which adopted a canonical DsbA fold similar to that observed in the structures of NmDsbA3 and Escherichia coli DsbA (EcDsbA). Structural comparisons revealed variations around the active site and candidate peptide-binding region. Additionally, we demonstrate that all three NmDsbAs are strong oxidases with similar redox potentials; however, they differ from EcDsbA in their ability to be reoxidized by E. coli DsbB. Collectively, our studies suggest that the small structural differences between the NmDsbA enzymes and EcDsbA are functionally significant and are the likely determinants of substrate specificity. PMID- 19815020 TI - Aggression in rainbow trout is inhibited by both MR and GR antagonists. AB - The present study has investigated the effect of exogenous cortisol on aggression in juvenile rainbow trout, along with the involvement of mineralocorticoid (MR) and glucocorticoid receptors (GR) mediating the effects of cortisol. Fish were fed pellets supplemented with cortisol, the GR antagonist mifepristone (RU486) in combination with cortisol, the MR antagonist spironolactone (SA) in combination with cortisol or both antagonists in combination with cortisol. Aggressive behaviour was then assessed 1h subsequent to feeding. Our results showed that the attack latency was increased by exogenous cortisol, an effect that was not abolished by the antagonists. The intensity of aggression was not changed by exogenous cortisol. However, the intensity of aggression was significantly reduced by both antagonists. These results are discussed with regard to cortisol affecting aggressive behaviour through genomic and non-genomic pathways. Our results have demonstrated the involvement of both MR and GR in regulating behavioural responses during social interactions in teleost fish. The intensity of aggression seen in control and cortisol treated fish is probably mediated by the basal levels of cortisol through the intracellular MRs and GRs. We conclude that the initiative to engage in social confrontations is mediated through a non genomic pathway, which could involve extracellular corticoid receptors. Further, the majority of arguments lean towards the MR and GR antagonists blocking the effect of cortisol on aggressive intensity through intracellular receptors. If this is the case, then it is probable that these two aspects of aggressive behaviour are based on different neuronal mechanisms. PMID- 19815021 TI - Effect of moderate intake of sweeteners on metabolic health in the rat. AB - The rise in prevalence of obesity, diabetes, metabolic syndrome, and fatty liver disease has been linked to increased consumption of fructose-containing foods or beverages. Our aim was to compare the effects of moderate consumption of fructose containing and non-caloric sweetened beverages on feeding behavior, metabolic and serum lipid profiles, and hepatic histology and serum liver enzymes, in rats. Behavioral tests determined preferred (12.5-15%) concentrations of solutions of agave, fructose, high fructose corn syrup (HFCS), a combination of HFCS and Hoodia (a putative appetite suppressant), or the non-caloric sweetener Stevia (n=5/gp). HFCS intake was highest, in preference and self-administration tests. Groups (n=10/gp) were then assigned to one of the sweetened beverages or water as the sole source of liquid at night (3 nights/wk, 10wks). Although within the normal range, serum cholesterol was higher in the fructose and HFCS groups, and serum triglycerides were higher in the Agave, HFCS, and HFCS/Hoodia groups (vs. water-controls, p<0.05). Liver histology was normal in all groups with no evidence of steatosis, inflammation, or fibrosis; however serum alanine aminotransferase was higher in the fructose and HFCS groups (vs. water-controls, p<0.05). Serum inflammatory marker levels were comparable among Stevia, agave, fructose, HFCS, and water-consuming groups, however levels of IL-6 were significantly lower in association with the ingestion of Hoodia. There were no differences in terminal body weights, or glucose tolerance assessed by 120-min IVGTTs performed at the end of the 10-week regimen. We conclude that even moderate consumption of fructose-containing liquids may lead to the onset of unfavorable changes in the plasma lipid profile and one marker of liver health, independent of significant effects of sweetener consumption on body weight. PMID- 19815022 TI - Psychophysical correlates in children with sensory modulation disorder (SMD). AB - Sensory modulation disorder (SMD), affecting approximately 5% of children, is characterized by sensory over or under-responsiveness to a range of stimuli in several modalities. Children with over-responsiveness (SOR) demonstrate increased aversion to certain natural stimuli that manifests as increased distress and avoidance behaviors to common stimuli, accompanied by abnormal electrodermal responses and brain evoked potentials to various stimuli. This study is the first to use quantitative sensory testing to characterize the somatosensory sub modalities of children with SMD. Seventy eight children aged 6-10 years (44 SMD children and 34 classmate controls) were tested. A diagnosis of SMD and SMD-free using the Short Sensory Profile was ascertained by the Sensory Profile Questionnaire, both completed by participants' mothers. Sensory detection thresholds for skin warming, cooling, punctate dynamic tactile sensation, vibration and thermal pain thresholds for heat and cold were determined at several body sites. Pain and prickle intensities for pinprick and prickly stimuli and the duration and intensity of the after-sensations of prickliness and pain evoked by the prickle stimuli were assessed. Compared to the control children, SMD children showed significant cool hypoesthesia, higher pain intensity to pinprick and to prickly stimuli, and significantly more pain after-sensation to the prickly stimuli. No significant differences between groups were found in most of the sensory and pain thresholds at any tested site. These results indicate, for the first time, that children with SMD perceive more pain, and that their pain lasts longer. Our results demonstrate that SOR does not imply lowered sensory thresholds but abnormal processing suprathreshold noxious stimuli. PMID- 19815023 TI - Effect of sampling array irregularity and window size on the discrimination of sampled gratings. AB - The effect of sampling irregularity and window size on orientation discrimination was investigated using discretely sampled gratings as stimuli. For regular sampling arrays, visual performance could be accounted for by a theoretical analysis of aliasing produced by undersampling. For irregular arrays produced by adding noise to the location of individual samples, the incidence of perceived orientation reversal declined and the spatial frequency range of flawless performance expanded well beyond the nominal Nyquist frequency. These results provide a psychophysical method to estimate the spatial density and the degree of irregularity in the neural sampling arrays that limit human visual resolution. PMID- 19815024 TI - Differential changes in perceived contrast following contrast adaptation in humans. AB - Perceived contrast is reduced after prolonged exposure to a textured pattern (contrast adaptation). The size of this effect is dependent on the relationship between the adapting contrast and the test contrast. It is generally accepted that the greatest reductions occur when the adapting contrast is much higher than the test contrast. Here this relationship was examined for a wide range of spatial frequencies. The results show that the effect of the adapt/test ratio on perceived contrast following contrast adaptation is highly spatial frequency dependent. At high spatial frequencies >1cpd perceived contrast was reduced for all adapting contrasts, which is consistent with other studies. However, at low spatial frequencies (<1cpd) the perceived contrast was actually above veridical perception when the adapting contrast was lower than the test contrast. This finding has not been previously reported and has important implications for models of contrast perception. PMID- 19815025 TI - Active travel to school, BMI and participation in organised and non-organised physical activity among Portuguese adolescents. AB - OBJECTIVES: To assess the relationship between participation in organised (OPA) and non-organised physical activity (NOPA), commuting to school and BMI. METHODS: Questionnaire assessing participation in OPA, NOPA, and travel behavior submitted in spring 2006 to 721 Portuguese secondary school students. Active travellers walked or biked (AT). Passive travellers drove or took the bus (PT). Height and weight were measured. RESULTS: AT was related to NOPA among boys (odds ratio (OR)=1.8, p < or = 0.05) but not among girls. AT was unrelated to OPA. BMI was weakly associated with OPA (boys: OR=0.56, p < or = 0.05; girls: OR=0.54, p < or = 0.05) and not with NOPA. CONCLUSION: After-school organised and non-organised physical activities do not predict active travelling to school among Portuguese adolescents. PMID- 19815026 TI - Structural adaptation of serine hydroxymethyltransferase to low temperatures. AB - Structural adaptation of serine hydroxymethyltransferase (SHMT), a pyridoxal-5' phosphate dependent enzyme that catalyzes the reversible conversion of l-serine and tetrahydropteroylglutamate to glycine and 5,10-methylene tetrahydropteroylglutamate, synthesized by microorganisms adapted to low temperatures has been analyzed using a comparative approach. The variations of amino acid properties and frequencies among three temperature populations (psychrophilic, mesophilic, hyper- and thermophilic) of SHMT sequences have been tested. SHMTs display a general increase of polarity specially in the core, a more negatively charged surface, and enhanced flexibility. Subunit interface is more hydrophilic and less compact. Electrostatic potential of the tetrahydrofolate binding site has been compared. The enzyme from Psychromonas ingrahamii, the organism with the lowest adaptation temperatures, displayed the most positive potential. In general, the property variations show a coherent opposite trend in the hyperthermophilic population: in particular, increase of hydrophobicity, packing and decrease of flexibility was observed. PMID- 19815027 TI - Novel targets for anti-retroviral therapy. AB - Novel targets for the management of HIV infection have become increasingly relevant in view of extensive drug resistance, side effects and high pill burden of some of the conventional anti-retroviral agents. Structure based drug design using X-ray crystallography, nuclear magnetic resonance, and mass spectrometry have lead to the identification of novel class of drugs targeting new stages of HIV life cycle. These agents include chemokine receptor antagonists and the integrase inhibitors which were recently approved for HIV treatment, as well as numerous other agents directed to previously untested targets such as the maturation inhibitors, zinc finger inhibitors, pharmacological CDK inhibitors, Tat-TAR interaction inhibitors, anti-CD4 monoclonal antibody, antisense oligonucleotides, oxidisers of the HIV lipid envelope and agents acting on the proviral DNA. Use of new agents with novel mechanism of action requires the development of new laboratory assays to detect viral tropism and new resistance mutations. Despite the new developments providing hope to patients and clinicians in the fights against HIV, eradication of the disease still remains elusive. This review discusses issues surrounding the development of these new agents. PMID- 19815028 TI - Lipopolysaccharide: Biosynthetic pathway and structure modification. AB - Lipopolysaccharide that constitutes the outer leaflet of the outer membrane of most Gram-negative bacteria is referred to as an endotoxin. It is comprised of a hydrophilic polysaccharide and a hydrophobic component referred to as lipid A. Lipid A is responsible for the major bioactivity of endotoxin, and is recognized by immune cells as a pathogen-associated molecule. Most enzymes and genes coding for proteins responsible for the biosynthesis and export of lipopolysaccharide in Escherichia coli have been identified, and they are shared by most Gram-negative bacteria based on genetic information. The detailed structure of lipopolysaccharide differs from one bacterium to another, consistent with the recent discovery of additional enzymes and gene products that can modify the basic structure of lipopolysaccharide in some bacteria, especially pathogens. These modifications are not required for survival, but are tightly regulated in the cell and closely related to the virulence of bacteria. In this review we discuss recent studies of the biosynthesis and export of lipopolysaccharide, and the relationship between the structure of lipopolysaccharide and the virulence of bacteria. PMID- 19815029 TI - Importance of normothermia control in investigating delayed neuronal injury in a mouse global ischemia model. AB - This study aims to establish a mouse global cerebral ischemia model in which the physiological parameter measurements and neuronal injury evaluations are conducted in the same group of animals and to identify the effect of post ischemic core temperature (CT) on the outcome of neuronal injury. Global ischemia was induced by 12-min bilateral common carotid artery occlusion followed by 7 days of reperfusion in C57BL/6 mice. Immediately after occlusion, mice were randomly assigned to be kept in environments of different temperatures [25 degrees C (room temperature, group 1), 33-34 degrees C for 2h (group 2), and 33 34 degrees C for 24h (group 3)] before being returned to their home cages. We found that in group 1, CT declined to approximately 32 degrees C after ischemia and then recovered at 24h post-ischemia; in group 2, CT remained at the pre ischemia level during the first 2h, declined after the mice were returned to room temperature, and recovered at 24h post-ischemia; and in group 3, CT remained constant at the pre-ischemia level throughout the reperfusion period. The number of surviving neurons in a sector of the hippocampal CA1 region was significantly lower in all ischemic groups than in the sham controls, but the number was significantly higher in group 1 than that in groups 2 or 3 (P<0.05). We observed that CT declines initially but recovers spontaneously at 24h post-ischemia. Early post-ischemic hypothermia impacts the delayed neuronal injury, suggesting that tight temperature control immediately following ischemia is important to obtain the most reproducible neuronal damage in mouse models of cerebral global ischemia. PMID- 19815030 TI - Development of a real-time RT-PCR assay for a novel influenza A (H1N1) virus. AB - A pandemic caused by a novel influenza A virus (H1N1) poses a serious public health threat. In this study, a real-time reverse transcriptase PCR (RT-PCR) assay based on the hemagglutinin gene was developed that discriminates the novel H1N1 from swine influenza virus, seasonal H1N1/H3N2 virus and the highly pathogenic H5N1 avian influenza virus. The sensitivity of this assay was 0.2 50% tissue culture infective dose of virus and 200 copies of in vitro-transcribed target RNA. Three hundred and forty-eight clinical specimens from suspected H1N1 patients were tested using this assay, and forty-two (12.07%) were found to be positive. Tests using the real-time PCR assay recommended by WHO and virus isolation gave identical results. This sensitive and specific real-time RT-PCR assay will contribute to the early diagnosis and control of the emerging H1N1 influenza pandemic. PMID- 19815031 TI - Development and laboratory validation of a lateral flow device for the detection of serotype SAT 2 foot-and-mouth disease viruses in clinical samples. AB - A lateral flow device (LFD) for the detection of foot-and-mouth disease virus (FMDV) of the SAT 2 serotype was developed using a monoclonal antibody (Mab 2H6). The performance of the LFD was evaluated in the laboratory on suspensions of vesicular epithelia: 305 positive for FMDV type SAT 2 from suspected cases of vesicular disease collected from 30 countries and 1002 samples shown to be negative for FMDV type SAT 2 collected from 67 countries between 1968 and 2008. The diagnostic sensitivity of the LFD for FMDV type SAT 2 was higher at 88% compared to 79% obtained by the reference method of antigen ELISA, and the diagnostic specificity of the LFD was approximately 99% compared to 100% for the ELISA. The device recognized FMDV strains of wide diversity within the FMDV SAT 2 serotype and gave a superior performance for their detection compared to the 1F10 LFD which had been developed previously and shown to perform less well for the detection of FMDVs of this particular serotype. Reactions in the SAT 2 2H6 LFD with the viruses of other FMDV serotypes and swine vesicular disease (which produces a clinically indistinguishable syndrome in pigs), did not occur. These data illustrate the potential for the LFD to be employed to complement the 1F10 device next to the animal in the pen-side diagnosis of FMD, for providing rapid and objective support to veterinarians in their clinical judgment of the disease and for specific confirmation of a FMDV type SAT 2 infection. PMID- 19815032 TI - Antimanic efficacy of retigabine in a proposed mouse model of bipolar disorder. AB - Retigabine is a novel compound with anticonvulsant efficacy. Preclinical studies have indicated that the compound, like other anticonvulsants may also have antimanic efficacy. Bipolar disorder is characterized by episodes of depression and mania, which show a progressively faster recurrence and an increase in severity with time. Recurrence of episodes in bipolar disorders is suggested to reflect a process of sensitization. Repeated intermittent administration of amphetamine in rodents gives rise to a behavioral sensitization phenomena argued to have similarities to the sensitization found in humans. The aims were therefore to explore the predictive validity of the amphetamine sensitization model as a behavioral model of mania by testing the effect of a range of antimanic drugs and to evaluate the effect of retigabine on the sensitized amphetamine response. Furthermore, since withdrawal from prolonged use of amphetamine in humans can result in depression symptoms it was explored if a state of anhedonia could be assessed by testing saccharine preference before and during the withdrawal period of the model. The tested antimanic drugs (lithium, valproate, carbamazepine and lamotrigine) all attenuated the sensitized locomotor activity induced and with the exception of valproate the found effects seemed not to be due to sedation. Interestingly, retigabine also attenuated the induced locomotor activity with a lowest effective dose at 1.0mg/kg, whereas basal locomotor activity was only reduced at 8.0mg/kg, suggesting a genuine calming and antimanic-like efficacy of the compound. In addition, saccharine preference data suggest that withdrawal from the d-amphetamine pre-treatment regimen may induce depression-like behavior indicating that both manic and depression-like behavior is expressed in this mouse model. PMID- 19815033 TI - Validation of a new method for immobilising kinetoplastid parasites for live cell imaging. AB - The kinetoplastid parasites are responsible for three of the ten most neglected tropical diseases as classified by the WHO. Recent advances in molecular and cellular analyses have allowed rapid progress in our understanding of the biology of these lethal pathogens. In this study we validate a new method for immobilising Trypanosoma brucei and Leishmania major parasites while maintaining a high level of viability. This allows reproducible live cell imaging of these highly motile organisms, thus enabling a full complement of advanced microscopic techniques to be utilised to better understand these pathogenic species. PMID- 19815035 TI - Review of brain functioning in depression for semantic processing and verbal fluency. AB - Neurobiological models of depression point to brain regions that are proposed to be involved with both emotion regulation and language processing. This qualitative review focused on neurophysiological evidence for semantic processing and verbal fluency deficits associated with left frontal lobe and dorsolateral prefrontal cortex functioning in depression, respectively. Findings suggest that there are no behavioral or neurophysiological evidence of performance differences between depressed and healthy individuals for semantic processing of neutral information, arguing against generalized left frontal lobe deficits. However, the preponderance of evidence points to enhanced processing of negative information in both left and right frontal lobes and behavior. Studies of verbal fluency were limited to non-emotional information. The majority of studies evaluated phonemic verbal fluency in depression (e.g., producing words that begin with a particular letter) and results generally showed bilateral hypoactivation of the frontal lobe with no concomitant deficits in behavioral performance. Overall, semantic processing and verbal fluency studies did not provide substantive evidence of specific left frontal lobe deficits. Evidence that emotional information may differentially impact brain functioning relative to neutral information in depression suggests that examination of verbal fluency for emotional information may contribute to the elucidation of executive functioning processes associated with the dorsolateral prefrontal cortex in depression. PMID- 19815036 TI - Nanovehicles for enhanced oral delivery of taxanes. PMID- 19815037 TI - Differential metabolic responses to pluronic in MDR and non-MDR cells: a novel pathway for chemosensitization of drug resistant cancers. AB - A synthetic amphiphilic block copolymer, Pluronic, is a potent chemosensitizer of multidrug resistant (MDR) cancers that has shown promise in clinical trials. It has unique activities in MDR cells, which include a decrease in ATP pools and inhibition of P-glycoprotein (Pgp) resulting in increased drug accumulation in cells. This work demonstrates that Pluronic rapidly (15min) translocates into MDR cells and co-localizes with the mitochondria. It inhibits complex I and complex IV of the mitochondria respiratory chain, decreases oxygen consumption and causes ATP depletion in MDR cells. These effects are selective and pronounced for MDR cells compared to non-MDR counterparts and demonstrated for both drug-selected and Pgp-transfected cell models. Furthermore, inhibition of Pgp functional activity also abolishes the effects of Pluronic on intracellular ATP levels in MDR cells suggesting that Pgp contributes to increased responsiveness of molecular "targets" of Pluronic in the mitochondria of MDR cells. The Pluronic caused impairment of respiration in mitochondria of MDR cells is accompanied with a decrease in mitochondria membrane potential, production of ROS, and release of cytochrome c. Altogether these effects eventually enhance drug-induced apoptosis and contribute to potent chemosensitization of MDR tumors by Pluronic. PMID- 19815038 TI - Temperature-dependent transfer of amphotericin B from liposomal membrane of AmBisome to fungal cell membrane. AB - Liposomal amphotericin B (AMPH-B), also known as AmBisome, exhibits a potent antifungal effect through its binding to ergosterol contained within the fungal cell membrane. However, the mechanism responsible for the movement of AmBisome derived AMPH-B to the fungal cell membrane through the cell wall is not yet clear. Therefore, in the present study we aimed at elucidating this mechanism operating in Saccharomyces cerevisiae. AmBisome showed its antifungal effect against S. cerevisiae at 35 degrees C but not at 4 degrees C, whereas free AMPH-B was effective at both temperatures. A significant difference in the amount of AMPH-B transferred to the fungal cells between incubation at 4 and 35 degrees C was also observed when AmBisome was used. Confocal microscopic study, however, indicated that NBD-labeled AmBisome was localized on the surface of the fungal cells at either temperature. To decrease the affinity of AMPH-B for the liposomal membrane, we entrapped AMPH-B in fluid liposomes containing egg yolk phosphatidylcholine (EPC) instead of hydrogenated soy PC (HSPC). These liposomes showed the antifungal effect even at 4 degrees C. On the contrary, AMPH-B in liposomes containing ergosterol (Erg-AmB) instead of cholesterol showed a significantly weaker antifungal effect at 35 degrees C with reduced transfer of AMPH-B to the fungal cells. These results suggest that not the binding of AmBisome to target cells but the transfer of AMPH-B from liposomal membrane of AmBisome to the cell membrane is critical for the antifungal activity of AmBisome. This transfer is dependent on the temperature, fluidity of the liposomal membrane, and the affinity of AMPH-B for the fungal cell membrane. PMID- 19815039 TI - How to study dendriplexes II: Transfection and cytotoxicity. AB - This paper reviews different techniques for analyzing the transfection efficiencies and cytotoxicities of dendriplexes-complexes of nucleic acids with dendrimers. Analysis shows that three plasmids are mainly used in transfection experiments: plasmid DNA encoding luciferase from the firefly Photinus pyralis, beta-galactosidase, or green fluorescent protein. The effective charge ratio of transfection does not directly correlate with the charge ratio obtained from gel electrophoresis, zeta-potential or ethidium bromide intercalation data. The most popular cells for transfection studies are human embryonic kidney cells (HEK293), mouse embryonic cells (NIH/3T3), SV40 transformed monkey kidney fibroblasts (COS 7) and human epithelioid cervical carcinoma cells (HeLa). Cellular uptake is estimated using fluorescently-labeled dendrimers or nucleic acids. Transfection efficiency is measured by the luciferase reporter assay for luciferase, X-Gal staining or beta-galactosidase assay for beta-galactosidase, and confocal microscopy for green fluorescent protein. Cytotoxicity is determined by the MTT test and lactate dehydrogenase assays. On the basis of the papers reviewed, a standard essential set of techniques for characterizing dendriplexes was constructed: (1) analysis of size and shape of dendriplexes in dried/frozen state by electron or atomic force microscopy; (2) analysis of charge/molar ratio of complexes by gel electrophoresis or ethidium bromide intercalation assay or zeta potential measurement; (3) analysis of hydrodynamic diameter of dendriplexes in solution by dynamic light scattering. For the evaluation of transfection efficiency the essential techniques are (4) luciferase reporter assay, beta galactosidase assay or green fluorescent protein microscopy, and (5) cytotoxicity by the MTT test. All these tests allow the transfection efficiencies and cytotoxicities of different kinds of dendrimers to be compared. PMID- 19815040 TI - Predicting food hygiene. An investigation of social factors and past behaviour in an extended model of the Health Action Process Approach. AB - PURPOSE: The current study uses an extended version of the Health Action Process Approach (HAPA) to predict food safety behaviour. DESIGN/METHODOLOGY/APPROACH: Two hundred and fifty-nine participants completed a questionnaire assessing variables of HAPA as well as additional predictors including: risk awareness, outcome expectancy, action self-efficacy, subjective norm, social support, past behaviour, and intention. One week later, participants returned a follow up questionnaire measuring their planning, maintenance self-efficacy, recovery self efficacy, social support and behaviour. Regressions were utilised to predict variances in intention, planning and behaviour. FINDINGS: The original HAPA model predicted 30.8% of the variance in intention and 17% of the variance in behaviour, confirming the suitability of the HAPA to predict food safety behaviour. The HAPA constructs also predicted 17.2% of variance in planning. The extended version of the model significantly increased the proportion of variance explained to 54.3% and 38.8% of variance in intention and behaviour respectively. RESEARCH LIMITATIONS/IMPLICATIONS: The HAPA was able to predict intention and behaviour comparably to other studies. The additional variables revealed intention to adopt food safety behaviours is partially dependent on parents of individuals, and behaviour is habitual in nature, explaining the low predictive power of planning. PRACTICAL IMPLICATIONS: The current study sheds light on the behaviour and suggests possible ways to design interventions. Interventions should focus on the habitual nature of the behaviour, thus implementing new correct habits. ORIGINALITY/VALUE: This study aimed to bolster the lack of studies using the HAPA to model food safety behaviour. This study also aimed to include additional variables into HAPA to increase the amount of predicted variance in intention and behaviour. PMID- 19815041 TI - Willingness-to-accept and purchase genetically modified rice with high folate content in Shanxi Province, China. AB - Neural-tube defects (NTDs) are considered to be the most common congenital malformations. As Shanxi Province, a poor region in the North of China, has one of the highest reported prevalence rates of NTDs in the world, folate fortification of rice is an excellent alternative to low intake of folate acid pills in this region. This paper investigates the relations between socio demographic indicators, consumer characteristics (knowledge, consumer perceptions on benefits, risks, safety and price), willingness-to-accept and willingness-to pay genetically modified (GM) rice. The consumer survey compromises 944 face-to face interviews with rice consumers in Shanxi Province, China. Multivariate analyses consist of multinomial logistic regression and multiple regression. The results indicate that consumers generally are willing-to-accept GM rice, with an acceptance rate of 62.2%. Acceptance is influenced by objective knowledge and consumers' perceptions on benefits and risks. Willingness-to-pay GM rice is influenced by objective knowledge, risk perception and acceptance. Communication towards the use of GM rice should target mainly improving knowledge and consumers' perceptions on high-risk groups within Shanxi Province, in particular low educated women. PMID- 19815034 TI - Identification of immediate response genes dominantly expressed in juvenile resistant and susceptible Biomphalaria glabrata snails upon exposure to Schistosoma mansoni. AB - Resistance or susceptibility of the snail host Biomphalaria glabrata to Schistosoma mansoni is determined by the genetics of both the snail and parasite. Although Mendelian genetics governs adult resistance to infection, juvenile resistance and susceptibility are complex traits. In this study, suppression subtractive hybridization was used to construct forward and reverse cDNA libraries to identify genes involved in the immediate response of juvenile resistant (BS-90), non-susceptible (LAC2) snails, and susceptible (NMRI) snails after early exposure to S. mansoni. Expressed Sequence Tags (ESTs) were generated from the repertoire of enriched transcripts. In resistant snails, several ESTs corresponded to transcripts involved in immune regulation/defense response. While no defense related transcripts were found among juvenile susceptible snail ESTs, we detected transcripts involved in negative regulation of biological process/morphogenesis/proliferation. Differential gene expression and temporal regulation of representative transcripts were compared among snails pre- and post exposure to either normal or attenuated miracidia using quantitative real time RT PCR. Results showed that several transcripts, such as fibrinolytic C terminal domain, cytidine deaminase, macrophage expressed gene 1, protein kinase C receptor, anti-microbial peptide; theromacin and Fas remained up-regulated regardless of whether or not snails were exposed to normal or attenuated miracidia. While ESTs related to C-type lectin and low-density lipoprotein receptor were induced only by exposure to normal miracidia. By comparing changes in gene expression between resistant and susceptible juvenile snails responding either to normal or attenuated parasites, we can conclude that the transcription of genes associated with the intra-dermal penetration process of the snail host by invading miracidia may need to be taken into account when assessing differential gene expression between resistant and susceptible strains of B.glabrata in relation to S. mansoni exposure. PMID- 19815042 TI - Improved appetite after multi-micronutrient supplementation for six months in HIV infected South African children. AB - The aim of the study was to assess the effect of multi-micronutrient supplementation on the appetite of HIV-infected children. HIV-infected children (6-24 months) who had previously been hospitalized were enrolled into a double blind randomized trial, and given daily multi-micronutrient supplements or placebos for six months. Appetite tests were performed at enrollment and after three and six months. Appetite was measured as ad libitum intake of a commercial cereal test food served after an overnight fast according to standardized procedures. Body weights and total amount of test food eaten were measured. In total, 99 children completed the study (50 on supplements and 49 on placebos). Amounts eaten per kilogram body weight in the supplement group at enrollment and after six months were 36.7+/-17.7 g/kg (mean+/-SD) and 41.3+/-15.0 g/kg respectively, while the amounts in the placebo group were 47.1+/-14.9 g/kg and 45.7+/-13.1g/kg respectively. The change in amount eaten per kilogram body weight over six months was significantly higher in the supplement group (4.7+/-14.7 g/kg) than in the placebo group (-1.4+/-15.1g/kg). Multi-micronutrient supplementation for six months seems to significantly improve the appetite of HIV infected children. PMID- 19815043 TI - Negative mood increases selective attention to food cues and subjective appetite. AB - Following negative reinforcement and affect-regulation models of dysfunctional appetitive motivation, this study examined the effect of negative mood on objective and subjective cognitive indices of motivation for food; i.e., attentional bias for food cues and self-reported hunger/urge to eat, respectively. The study extended previous research on the effect of mood on food motivation by using (i) an experimental mood manipulation, (ii) an established index of attentional bias from the visual-probe task and (iii) pictorial food cues, which have greater ecological validity than word stimuli. Young female adults (n=80) were randomly allocated to a neutral or negative mood induction procedure. Attentional biases were assessed at two cue exposure durations (500 and 2000ms). Results showed that negative mood increased both attentional bias for food cues and subjective appetite. Attentional bias and subjective appetite were positively inter-correlated, suggesting a common mechanism, i.e. activation of the food-reward system. Attentional bias was also associated with trait eating style, such as external and restrained eating. Thus, current mood and trait eating style each influenced motivation for food (as reflected by subjective appetite and attentional bias). Findings relate to models of cognitive mechanisms underlying normal and dysfunctional appetitive motivation and eating behaviour. PMID- 19815044 TI - Emotional and uncontrolled eating styles and chocolate chip cookie consumption. A controlled trial of the effects of positive mood enhancement. AB - The study tested the effects of positive mood enhancement on chocolate chip cookie consumption in the context of emotional and uncontrolled eating styles. The relationship between emotional eating style and chocolate chip cookie intake was assumed to be mediated by uncontrolled eating style. Further, it was hypothesized that the effectiveness of the positive mood enhancement may be more salient among those who have effective control of their eating. In this experimental study, respondents (N=106, 70% women, aged 16-45 years old) were assigned by means of cluster randomization to the control or positive mood enhancement condition (a comedy movie clip). Compared to the control condition, positive mood enhancement resulted in consuming on average 53.86 kcal less. Relationships between emotional eating style and cookie intake were mediated by uncontrolled eating. Moderated mediation analysis indicated that the effect of a mediator (uncontrolled eating) on cookie intake was moderated by the group assignment. Positive mood enhancement resulted in eating on average 3.3 cookies less among individuals with a more controlled eating style. By contrast, among those who presented uncontrolled eating, positive mood enhancement led to consuming an average of 1.7 cookies more. PMID- 19815045 TI - Functional activation of monoamine transporters by luteolin and apigenin isolated from the fruit of Perilla frutescens (L.) Britt. AB - Monoamine transporters playing major roles in regulating normal and abnormal synaptic activity are associated with various neuropsychological disorders. In spite of the discovery of a series of structurally different monoamine transporter antagonists for the therapy approach, no practical pharmaceutical can act as a transporter activator. Here, we isolated luteolin and apigenin from the fruit of Perilla frutescens (L.) Britt by using an activity-guided extraction technique, and proved that the two compounds possess actions of enhancing monoamine uptake either upon monoamine-transporter transgenic Chinese hamster ovary (CHO) cells or upon wild dopaminergic cell lines, with higher specificity for dopamine (DA) uptake than for norepinephrine (NE)- and serotonin (5HT) uptake, as well as with more potency and greater efficacy for luteolin than for apigenin. Further, in the transgenic cells, the principal NE/DA uptake activation by luteolin was significantly prevented by respective transporter inhibitor, and the transmitter-uptake-enhancing action was independent of its ligands, which is in support of the compounds as monoamine transporter activators. Furthermore, luteolin evoked a marked disinhibition of cocaine-targeted effect in CHO cells overexpressing dopamine transporter. Thus, luteolin and apigenin function as monoamine transporter activators, which would improve several hypermonoaminergic neuropsychological disorders, especially cocaine dependence, through up regulating monoamine transporter activity. PMID- 19815046 TI - Matrix metalloproteinase-3 contributes to vulnerability of the nigral dopaminergic neurons. AB - Dopamine(DA)rgic neurons are particularly vulnerable due to the presence of oxidative stress-inducing molecules such as DA, tetrahydrobiopterin, iron and tyrosine hydroxylase (TH). We have recently observed that matrix metalloproteinase-3 (MMP-3) is involved in degeneration of DArgic neurons. In the present study, we sought to explore the role of MMP-3 in DArgic neurons not exposed to apparent stress conditions. In 8-week-old male mice deficient of MMP-3 gene (MMP-3 KO), the total number of DArgic neurons in the substantia nigra was considerably higher than wild type (WT). Primary cultured mesencephalic neurons from MMP-3 KO showed higher [(3)H]DA uptake capability compared to that of WT. The number of TH-immunopositive neurons and the length of average dendritic branch were also greater. This appeared to be selective for the DArgic system, because [(3)H]GABA uptake and calbindin D-28K and MAP-2 immunoreactivities were unaltered. On the other hand, no differences were noted in the levels of the striatal DA, DOPAC and BH4 and TH protein between the KO and WT. Interestingly, TH immunodensity per cell was lower in the DArgic neurons of MMP-3 KO both in primary culture and in vivo, suggesting the presence of a compensatory mechanism. These results further indicate a role of MMP-3 in the demise of DArgic neurons and suggest MMP-3 as a candidate cellular target for neuroprotective therapy. PMID- 19815047 TI - Co-engagement of alpha(4)beta(1) integrin (VLA-4) and CD4 or CD8 is necessary to induce maximal Erk1/2 phosphorylation and cytokine production in human T cells. AB - The alpha(4)beta(1) integrin VLA-4 (very-late activation antigen-4) and the lineage-specific CD4 and CD8 receptors have been proposed as putative co stimulatory receptors on T cells. To assess the relative contribution of signaling through the TCR, CD28 and these accessory molecules, we activated human T cells using soluble antibodies recognizing all four of these T-cell receptor classes (CD3, CD28, CD4/CD8, and VLA-4), and we assessed the degree of activation using higher-order flow cytometry detecting intracellular Erk1/2 phosphorylation and production of IL-2 and IFN-gamma. We found that: (1) co-stimulation via CD4/CD8, in addition to CD28, is required for optimal T-cell activation; (2) VLA 4 binding consistently potentiates CD4(+) and CD8(+) T-cell activation; (3) augmentation of T-cell activation through VLA-4 binding is most pronounced following engagement of CD4/CD8. These results confirm that multiple signals, including VLA-4 engagement, are necessary for maximal T-cell activation beyond that induced via the TCR and CD28. PMID- 19815048 TI - Ginkgo biloba for attention-deficit/hyperactivity disorder in children and adolescents: a double blind, randomized controlled trial. AB - BACKGROUND: Although stimulants are highly effective in controlling the symptoms of Attention-Deficit/Hyperactivity Disorder (ADHD), some children will not respond to, or are intolerant of stimulants. Thus, the desire for safe and effective nonstimulant medications has risen during the past several years. Ginkgo biloba has been suggested in the treatment of dementia and memory impairment. We hypothesized that G.biloba would be beneficial for treatment of ADHD, and this could be evaluated in a double blind, randomized, parallel group comparison of G.biloba (Ginko T.D. Tolidaru, Iran) and methylphenidate. METHODS: Fifty outpatients (39 boys and 11 girls) with a DSM-IV-TR diagnosis of ADHD were study population of this trial. Subjects were recruited from an outpatient child and adolescent clinic for a 6 week double blind, randomized clinical trial. All study subjects were randomly assigned to receive treatment using tablet of Ginko T.D. at a dose of 80-120 mg/day depending on weight (80 mg/day for <30 kg and 120 mg/day for >30 kg) (group 1) or methylphenidate at a dose of 20-30 mg/day depending on weight (20 mg/day for <30 kg and 30 mg/day for >30 kg (group 2) for a 6 week double blind, randomized clinical trial. The principal measure of outcome was the Teacher and Parent ADHD Rating Scale- IV. Patients were assessed at baseline and at 21 and 42 days after the medication started. RESULTS: Significant differences were observed between the two groups on the Parent and Teacher Rating Scale scores. The changes at the endpoint compared to baseline were: -6.52+/-11.43 (mean+/-S.D.) and -15.92+/-11.44 (mean+/-S.D.) for Ginko T.D. and methyphenidate, respectively for Parent ADHD Rating Scale. The changes at the endpoint compared to baseline were: -0.84+/-6.79 (mean+/-S.D.) and -14.04+/-8.67 (mean+/-S.D.) for Ginko T.D. and methyphenidate, respectively for Teacher ADHD Rating Scale. The difference between the Ginko T.D. and methylphenidate groups in the frequency of side effects was not significant except for decreased appetite, headache and insomnia that were observed more frequently in the methylphenidate group. CONCLUSION: The results of this study suggest that administration of G.biloba was less effective than methylphenidate in the treatment of ADHD. PMID- 19815049 TI - Stem cell-derived hepatocytes and their use in toxicology. AB - Better prediction of safety risk and understanding of mechanism of action of drug candidates remain a major challenge in order to prevent late stage attrition. Continuous efforts are made to improve and develop new models, especially in some areas such as hepatotoxicity. Besides primary hepatocytes and transformed liver cell lines, stem cells either isolated from embryos or adult tissues or obtained by reprogramming somatic cells are emerging as a new potential source of unlimited numbers of hepatocytes. Presently, only hepatocyte-like cells expressing low levels of liver-specific markers, especially drug metabolizing and detoxifying enzymes, are usually obtained, making them still unsuitable as metabolically competent cells for toxicity studies. The only exceptions are some hepatoma cell lines, particularly the HepaRG cell line that can differentiate from a bipotent progenitor stage to attain the functional capacity of normal adult hepatocytes in primary culture without losing the indefinite growth property of transformed cells. Since the research field on stem cells is growing fast marked advances might be expected in the next future. PMID- 19815050 TI - Polycythemia vera erythroid precursors exhibit increased proliferation and apoptosis resistance associated with abnormal RAS and PI3K pathway activation. AB - OBJECTIVE: Polycythemia vera (PV) is characterized by erythrocytosis associated with the presence of the activating JAK2(V617F) mutation in a variable proportion of hematopoietic cells. JAK2(V617F) is detected in other myeloproliferative neoplasms, does not appear to be the PV-initiating event, and its specific role in deregulated erythropoiesis in PV is incompletely understood. We investigated the pathogenesis of PV to characterize abnormal proliferation and apoptosis responses and aberrant oncogenic pathway activation in primary PV erythroid precursors. MATERIALS AND METHODS: Peripheral blood CD34(+) cells isolated from PV patients and healthy controls were grown in liquid culture to expand a population of primary erythroblasts for experiments designed to analyze cellular proliferation, apoptosis, JAK2(V617F) mutation status, cytokine-dependent protein phosphorylation and gene expression profiling using Affymetrix microarrays. RESULTS: The survival and proliferation of PV erythroblasts were growth factor dependent under strict serum-free conditions requiring both erythropoietin (EPO) and stem cell factor. PV erythroblasts exhibited EPO hypersensitivity and enhanced cellular proliferation associated with increased EPO-mediated extracellular signal-regulated kinases 1 and 2 phosphorylation. EPO-induced AKT phosphorylation was observed in PV but not normal erythroblasts, an effect associated with apoptosis resistance in PV erythroblasts. Analysis of gene expression and oncogenic pathway activation signatures revealed increased RAS (p<0.01) and phosphoinositide-3 kinase (p<0.05) pathway activation in PV erythroblasts. CONCLUSION: Deregulated erythropoiesis in PV involves EPO hypersensitivity and apoptosis resistance of erythroid precursor cells associated with abnormally increased activation of RAS-ERK and phosphoinositide-3 kinase-AKT pathways. These data suggest that investigation of the mechanisms of abnormal RAS and phosphoinositide-3 kinase pathway activation in erythroblasts may contribute to our understanding of the molecular pathogenesis of PV. PMID- 19815051 TI - The rapamycin-derivative RAD001 (everolimus) inhibits cell viability and interacts with the Akt-mTOR-p70S6K pathway in human medullary thyroid carcinoma cells. AB - Over-expression of the proto-oncogene Akt/PKB has been demonstrated in some neuroendocrine tumor models. Akt may activate downstream proteins such as mTOR and p70S6K, inducing tumor proliferation. The rapamycin-derivative RAD001, everolimus, interacts with this pathway by antagonizing mTOR, but its effects on neuroendocrine tumors are largely unknown. We explored the mechanism of action of RAD001 on cell proliferation, hormonal secretion and on Akt/mTOR/p70S6K pathway activation, in a human medullary thyroid carcinoma (MTC) cell-line (TT) and in cells derived from human MTCs. Treatment with RAD001 significantly inhibited cell viability in a dose- and time-dependent fashion, and diminished phosphorylation of Akt downstream targets, mTOR and p70S6K, in both TT cell-line and cultured human MTCs. Akt phosphorylation was not affected by RAD001. RAD001 induced cell cycle arrest in the G(0)/G(1) phase in TT cells, but had no effect on apoptosis. Moreover, RAD001 did not affect calcitonin and carcinoembryonic antigen secretion in TT cells and in human MTCs. RAD001 seems to have potent anti-proliferative effect in human MTC cells, which suggest that clinical trials of this agent are of considerable interest. PMID- 19815052 TI - The ERK and PI3K signaling pathways mediate inhibition of insulin-like growth factor-1 receptor mRNA expression by somatostatin. AB - Somatostatins (SSs) are a structurally diverse family of peptide hormones that regulate various aspects of growth, development, and metabolism in vertebrates. Previously, we showed that SSs inhibit mRNA and functional expression of insulin like growth factor-1 receptors (IGFR1) in gill filaments of rainbow trout. In this study, we used trout gill filaments, which express in high abundance two distinct IGFR1s, IGFR1A and IGFR1B, to examine the mechanism(s) through which SSs exert their inhibitory effects on IGFR1 expression. SS-14, a predominat SS isoform, directly stimulated the phosphorylation of extracellular signal regulated kinase (ERK) and protein kinase B (Akt), a downstream target of phosphatidylinositol 3-kinase (PI3K), in filaments incubated in vitro. Activation of ERK and Akt by SS-14 was rapid, occuring within 5-10 min, and was concentration-dependent. The ERK pathway inhibitor, U0126, retarded SS-14 stimulated phosphorylation of ERK 1/2, whereas the PI3K inhibitor, LY294002, blocked SS-14-stimulated phosphorylation of Akt. SS-14-inhibited expression of IGFR1 mRNAs was blocked by both U0126 and LY294002. These data indicate that SS 14 inhibition of IGFR1 mRNA expression is mediated through the ERK and PI3K/Akt signaling pathways. PMID- 19815053 TI - Sulfonylureas uncouple glucose-dependence for GPR40-mediated enhancement of insulin secretion from INS-1E cells. AB - Activation of GPR40 is reported to enhance insulin secretion in the presence of glucose. We determined whether sulfonylureas could replace glucose for GPR40 mediated enhancement of insulin secretion and investigated underlying mechanisms using INS-1E cells. GW9508, a specific agonist of GPR40, significantly enhanced insulin secretion in the presence of high concentrations of glucose. In contrast, sulfonylureas increased insulin secretion in the absence of glucose. In the presence of sulfonylureas, activation of GPR40 significantly enhanced insulin secretion. The L-type calcium channel (LTCC) activator S-(-)-Bay K8644 also concentration-dependently increased insulin secretion in the absence of glucose. In the presence of 10 micromol/L S-(-)-Bay K8644, GW9508 significantly increased insulin secretion. On the other hand, the LTCC blocker nifedipine significantly inhibited insulin secretion mediated by either glucose, glipizide or glucose plus GW9508. Thus, sulfonylureas could replace glucose to support GPR40-mediated enhancement of insulin secretion, whereas blockage of LTCC reduced both glucose and sulfonylurea-mediated insulin secretion. PMID- 19815054 TI - Obesity and type 2 diabetes: slow down!--Can metabolic deceleration protect the islet beta cell from excess nutrient-induced damage? AB - Islet beta-cell dysfunction is a characteristic and the main cause of hyperglycaemia of Type 2 diabetes. Understanding the mechanisms that cause beta cell dysfunction will lead to better therapeutic outcomes for patients with Type 2 diabetes. Chronic fatty acid exposure of susceptible islet beta-cells causes dysfunction and death and this is associated with increased reactive oxygen species production leading to oxidative stress and increased endoplasmic reticulum stress. We present the hypothesis that metabolic deceleration can reduce both oxidative and endoplasmic reticulum stress and lead to improved beta cell function and viability when exposed to a deleterious fat milieu. This is illustrated by the C57BL/6J mouse which is characterised by reduced insulin secretion and glucose intolerance associated with a mutation in nicotinamide nucleotide transhydrogenase (Nnt) but is resistant to obesity induced diabetes. On the other hand the DBA/2 mouse has comparatively higher insulin secretion and better glucose tolerance associated with increased Nnt activity but is susceptible to obesity-induced diabetes, possibly as a result of increased oxidative stress. We therefore suggest that in states of excess nutrient load, a reduced ability to metabolise this load may protect both the function and viability of beta-cells. Strategies that reduce metabolic flux when beta-cells are exposed to nutrient excess need to be considered when treating Type 2 diabetes. PMID- 19815055 TI - Diabetes alters protein expression of hyperpolarization-activated cyclic nucleotide-gated channel subunits in rat nodose ganglion cells. AB - Vagal afferent neurons, serving as the primary afferent limb of the parasympathetic reflex, could be involved in diabetic autonomic neuropathy. Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are expressed in the vagal afferent neurons and play an important role in determining cell membrane excitation. In the present study, the protein expression and the electrophysiological characteristics of HCN channels were investigated in nodose ganglion (NG) afferent neurons (A-fiber and C-fiber neurons) from sham and streptozotocin (STZ)-induced diabetic rats. In the sham NG, HCN1, HCN3, and HCN4 were expressed in the A-fiber neurons; and HCN2, HCN3, and HCN4 were expressed in the C-fiber neurons. Compared to the sham NG neurons, diabetes induced the expression of HCN2 in the A-fiber neurons besides overexpression of HCN1 and HCN3; and enhanced the expression of HCN2 and HCN3 in C-fiber neurons. In addition, whole-cell patch-clamp data revealed diabetes also increased HCN currents in A-fiber and C-fiber neurons. However, we found that diabetes did not alter the total nodose afferent neuron number and the ratio of A-fiber/C-fiber neurons. These results indicate that diabetes induces the overexpression of HCN channels and the electrophysiological changes of HCN currents in the A- and C fiber nodose neurons, which might contribute to the diabetes-induced alteration of cell excitability in the vagal afferent neurons. PMID- 19815056 TI - Crocin and geniposide profiles and radical scavenging activity of gardenia fruits (Gardenia jasminoides Ellis) from different cultivars and at the various stages of maturation. AB - The major components of gardenia fruits are geniposide and water soluble pigment crocins. In this study, we investigate crocins and geniposide profiles of gardenia fruits from different cultivars and at the various stages of maturation. DPPH scavenging activity of gardenia fruits from different cultivars and at the various stages of fruit maturation was also assayed. Quantitative determination of crocins in the gardenia at the various stages of maturation revealed a significant increase when ripening. However, geniposide content was negatively correlated with ripening stages. A significant difference was observed when comparing crocin content of different gardenia from various cultivars and geniposide content also showed marked variety. Current study indicated no relationship between crocin and geniposide content in gardenia fruits at the various stages of maturation and DPPH radical scavenging activity. Data showed that, although crocins feature markedly less DPPH scavenging activity than gardenia ethanol extract, total crocin content of gardenias collected in various cultivars correlate, to a certain degree, with radical scavenging effects of the Chinese traditional medicine (r=0.75). PMID- 19815057 TI - A further application of the active time model to multiple concurrent variable interval schedules. AB - In this experiment we show that the active time model (ATM) accurately predicts probe data from multiple concurrent VI VI schedules. Subjects were trained under a concurrent VI 30-s VI 60-s and a concurrent VI 60-s VI 120-s schedule. Two types of unreinforced probes were then conducted. The first paired the two VI 60 s stimuli. These stimuli, while equivalent in their associated absolute rates of reinforcement, differed in their relative rates of reinforcement. The second probe paired the VI 30-s stimulus with the relatively rich VI 60-s stimulus. In contrast with the first probe, these stimuli differed in their absolute rates of reinforcement, while being similar in their relative rates. During the first set of probes, birds preferred the VI 60-s stimulus trained with the VI 120-s schedule. During the second set of probes, birds were indifferent to the two stimuli. These results are less extreme than others reported in the literature. Nonetheless, we found that ATM accurately fit individual subject data in both sets of probes. In contrast a variant of scalar expectancy theory did not fit the data at either the individual or group level. PMID- 19815058 TI - Posture in ovo as a precursor of footedness in ostriches (Struthio camelus). AB - Two categories of behaviour involving lateralized posture were observed in semi natural conditions in ostriches (Struthio camelus). Observing preferences for left or right foot, both in the forward foot posture (the foot standing in front at rest) and the starting foot used to initiate locomotion, a population-level right-foot preference was shown for the whole group and for each of the three age ranges considered (chick, young and adult). Ostriches are known to rely upon a lateralized behaviour during hatching (using their right foot to break the egg shell) suggesting the hypothesis that the precocious motor laterality observed at hatching might stand as a precursor of limb preference later in development, as already observed in other avian species. PMID- 19815059 TI - Site-specific bone loss in senescence-accelerated mouse (SAMP6): a murine model for senile osteoporosis. AB - The senescence-accelerated mouse strain P6 (SAMP6) is a model of senile osteoporosis, which possesses many features of senile osteoporosis in humans. So far, little is known about the systemic bone microstructural changes that occur at multiple skeletal sites. In this study, we therefore, investigated site (vertebra, femur and tibia) dependence of bone microstructure and bone mineral density (BMD) in SAMP6 and the normal control mouse (SAMR1) at 5 and 12months of age using quantitative micro computed tomography (micro-CT) and image analysis software. As compared with SAMR1, the most prominent change in SAMP6 was the reduction of vertebral trabecular bone volume fraction (BV/TV) and trabecular BMD. Moderate decrease of trabecular bone mass was observed in the proximal tibia and distal femur. Increased marrow area and periosteal perimeter were investigated, though the cortical area and cortical thickness had no marked changes in the mid-tibial and mid-femoral cortical bones. These results indicate that bone microstructural properties in SAMP6 are remarkably heterogeneous throughout the skeleton, which is analogous to changes that occur in human bones. These findings further validate the relevance of SAMP6 as a model of senile osteoporosis. PMID- 19815060 TI - Evaluation of transcriptional activity of caspase-3 gene as a marker of acute neurotoxicity in rat cerebellar granular cells. AB - Caspase-3 is a key protein involved in the classical apoptosis mechanism in neurons, as in many other cells types. In the present research, we describe the transcriptional activity of caspase-3 gene as a marker of acute toxicity in a primary culture model of rat cerebellar granule neurons (CGNs). CGNs were incubated for 16h in complete medium containing the chemicals at three concentrations (10, 100microM and 1mM). A total of 48 different compounds were tested. Gene transcriptional activity was determined by low-density array assays, and by single Taqman caspase-3 assays. Results from the PCR arrays showed that the caspase-3 gene was up-regulated when CGNs were exposed to neurotoxic chemicals. Significative correlations were found between the transcriptional activity of caspase-3 and the activity of some other genes related to apoptosis, cell-cycle and ROS detoxification. In our experiments, acute exposure of CGNs to well-documented pro-apoptotic xenobiotics modulated significantly caspase-3 gene expression, whereas chemicals not related to apoptosis did not modify caspase-3 gene expression. In conclusion, acute exposure of CGNs to neurotoxic compounds modulates the transcriptional activity of genes involved in the classical apoptotic pathway, oxidative stress and cell-cycle control. Transcriptional activity of caspase-3 correlates significantly with these changes and it could be a good indicator of acute neurotoxicity. PMID- 19815061 TI - Induction of oxidative stress and inflammatory cytokines by manganese chloride in cultured T98G cells, human brain glioblastoma cell line. AB - Manganese, an essential trace nutrient in human beings, has been widely used in the steel industry to improve hardness, stiffness, and strength. With the increased applications of manganese compounds, discharge into the environment has rapidly increased and may exert adverse effects on human health. In this study, manganese toxicity was investigated using cultured T98G cells, which are derived from human glioblasts with the ability to differentiate into several different types of neuroglia. Cytotoxicity was shown in manganese-treated groups (100, 200, 400, and 800microM of MnCl(2)), and cell viability was decreased to 58.8% of the control group at 2days after treatment with 800microM of MnCl(2). When cells were treated with manganese for 24h, ROS dose-dependently increased while antioxidant intracellular GSH decreased. With the generation of ROS, the increased activity of caspase-3 was shown, and was followed by chromatin condensation and breakage, which is an indication of the cellular apoptotic process. ROS also triggered pro inflammatory responses in cultured T98G cells, which were demonstrated by the increased gene expression and protein levels of IL-6 and IL-8. PMID- 19815062 TI - Methodology for phase selection of a weak basic drug candidate, utilizing kinetic solubility profiles in bio-relevant media. AB - We aimed to develop a phase selection methodology for a weak basic active pharmaceutical ingredient (API) that would require less than 10mg of the API and monitor the real-time kinetic solubility of the API in two bio-relevant media. Three sets of kinetic solubility measurements were conducted for free form I and the disulfate salt of an API (compound A) in order to determine the better API phase for further development of the compound. Tests consisted of solid API dissolution in both simulated gastric fluid (SGF) and fasted-state intestinal fluid (FaSSIF), and precipitation kinetics by injection of liquid state API into FaSSIF. All dissolution tests were conducted above the saturated concentrations in order to determine the compounds' thermodynamic and kinetic solubility to trace the API's phase transitions during dissolution. The pharmacokinetic profiles of compound A following oral administration of two API phases were evaluated in dogs. Results of the three sets of kinetic solubility measurements showed different kinetic solubility profiles for the two API phases under gastrointestinal conditions, indicating that the disulfate salt is preferred over free form I due to its superior kinetic solubility profile. This conclusion is consistent with the bioavailability results obtained in dogs. PMID- 19815063 TI - Artificial neural networks in the optimization of a nimodipine controlled release tablet formulation. AB - Artificial neural networks (ANNs) were employed in the optimization of a nimodipine zero-order release matrix tablet formulation, and their efficiency was compared to that of multiple linear regression (MLR) on an external validation set. The amounts of PEG-4000, PVP K30, HPMC K100 and HPMC E50LV were used as independent variables following a statistical experimental design, and three dissolution parameters (time at which the 90% of the drug was dissolved, t(90%), percentage of nimodipine released in 2 and 8h, Y(2h), and Y(8h), respectively) were chosen as response variables. It was found that a feed-forward back propagation ANN with eight hidden units showed better fit for all responses (R(2) of 0.96, 0.90 and 0.98 for t(90%), Y(2h) and Y(8h), respectively) compared to the MLR models (0.92, 0.87 and 0.92 for t(90%), Y(2h) and Y(8h), respectively). The ANN was further simplified by pruning, which preserved only PEG-4000 and HPMC K100 as inputs. Optimal formulations based on ANN and MLR predictions were identified by minimizing the standardized Euclidian distance between measured and theoretical (zero order) release parameters. The estimation of the similarity factor, f(2), confirmed ANNs increased prediction efficiency (81.98 and 79.46 for the original and pruned ANN, respectively, and 76.25 for the MLR). PMID- 19815064 TI - Estrogen utilization of IGF-1-R and EGF-R to signal in breast cancer cells. AB - As breast cancer cells develop secondary resistance to estrogen deprivation therapy, they increase their utilization of non-genomic signaling pathways. Our prior work demonstrated that estradiol causes an association of ERalpha with Shc, Src and the IGF-1-R. In cells developing resistance to estrogen deprivation (surrogate for aromatase inhibition) and to the anti-estrogens tamoxifen, 4-OH tamoxifen, and fulvestrant, an increased association of ERalpha with c-Src and the EGF-R occurs. At the same time, there is a translocation of ERalpha out of the nucleus and into the cytoplasm and cell membrane. Blockade of c-Src with the Src kinase inhibitor, PP-2 causes relocation of ERalpha into the nucleus. While these changes are not identical in response to each anti-estrogen, ERalpha binding to the EGF-R is increased in response to 4-OH-tamoxifen when compared with tamoxifen. The changes in EGF-R interactions with ERalpha impart an enhanced sensitivity of tamoxifen-resistant cells to the inhibitory properties of the specific EGF-R tyrosine kinase inhibitor, AG 1478. However, with long term exposure of tamoxifen-resistant cells to AG 1478, the cells begin to re-grow but can now be inhibited by the IGF-R tyrosine kinase inhibitor, AG 1024. These data suggest that the IGF-R system becomes the predominant signaling mechanism as an adaptive response to the EGF-R inhibitor. Taken together, this information suggests that both the EGF-R and IGF-R pathways can mediate ERalpha signaling. To further examine the effects of fulvestrant on ERalpha function, we examined the acute effects of fulvestrant, on non-genomic functionality. Fulvestrant enhanced ERalpha association with the membrane IGF-1-receptor (IGF-1-R). Using siRNA or expression vectors to knock-down or knock-in selective proteins, we further demonstrated that the ERalpha/IGF-1-R association is Src-dependent. Fulvestrant rapidly induced IGF-1-R and MAPK phosphorylation. The Src inhibitor PP2 and IGF-1 R inhibitor AG1024 greatly blocked fulvestrant-induced ERalpha/IGF-1-R interaction leading to a further depletion of total cellular ERalpha induced by fulvestrant and further enhanced fulvestrant-induced cell growth arrest. More dramatic was the translocation of ERalpha to the plasma membrane in combination with the IGF-1-R as shown by confocal microscopy. Taken in aggregate, these studies suggest that secondary resistance to hormonal therapy results in usage of both IGF-R and EGF-R for non-genomic signaling. PMID- 19815065 TI - H2O2-dependent translocation of TCTP into the nucleus enables its interaction with VDR in human keratinocytes: TCTP as a further module in calcitriol signalling. AB - Translationally controlled tumour protein (TCTP) is an evolutionarily highly conserved molecule implicated in many processes related to cell cycle progression, proliferation and growth, to the protection against harmful conditions including apoptosis and to the human allergic response. We are showing here that after application of mild oxidative stress, human TCTP relocates from the cytoplasm to the nuclei of HaCaT keratinocytes where it directly associates with the ligand-binding domain of endogenous vitamin D(3) receptor (VDR) through its helical domain 2 (AA 71-132). Interestingly, the latter harbours a putative nuclear hormone receptor coregulatory LxxLL-like motif which seems to be involved in the interaction. Moreover, we demonstrate that VDR transcriptionally induces the expression of TCTP by binding to a previously unknown VDR response element within the TCTP promotor. Conversely, ectopically overexpressed TCTP downregulates the amount of VDR on both mRNA as well as protein level. These data, to conclude, suggest a kind of feedback regulation between TCTP and VDR to regulate a variety of (Ca(2+) dependent) cellular effects and in this way further underscore the physiological relevance of this novel protein-protein interaction. PMID- 19815066 TI - Four and a half LIM domain 2 alters the impact of aryl hydrocarbon receptor on androgen receptor transcriptional activity. AB - Aryl hydrocarbon receptor (AhR) ligands modulate androgen receptor (AR) signaling in prostate cancer cells through partially defined mechanisms. Furthermore, these facilitatory and inhibitory effects of AhR on AR signaling appear to be cell or context specific. In the present study we demonstrate that both AhR and AhR nuclear translocator (ARNT) interact with AR. AhR but not ARNT enhanced the AR transcriptional activity which was independent of exogenous AhR ligand treatment (2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD). We then tested if coactivators common to both receptors alter the facilitatory effect of AhR on AR activity. NcoA4 overexpression did not alter the AhR facilitatory effect on AR, whereas SRC1 overexpression further enhanced the effect. In contrast, FHL2 overexpression blocked the facilitatory effect of AhR. In the presence of exogenous FHL2 expression, AhR repressed AR activity, whereas at low endogenous levels of FHL2 expression, AhR overexpression enhanced AR activity. At high FHL2 expression levels, TCDD treatment decreased AR activity and this effect was reversed by AhR overexpression. These findings demonstrate that AhR modulation of AR activity is differentially altered by the level of FHL2 and AhR present in the cell. PMID- 19815067 TI - Ingested double-stranded RNAs can act as species-specific insecticides. AB - A serious shortcoming of many insecticides is that they can kill non-target species. To address this issue, we harnessed the sequence specificity of RNA interference (RNAi) to design orally-delivered double-stranded (ds) RNAs that selectively killed target species. Fruit flies (Drosophila melanogaster), flour beetles (Tribolium castaneum), pea aphids (Acyrthosiphon pisum), and tobacco hornworms (Manduca sexta) were selectively killed when fed species-specific dsRNA targeting vATPase transcripts. We also demonstrate that even closely related species can be selectively killed by feeding on dsRNAs that target the more variable regions of genes, such as the 3' untranslated regions (UTRs): four species of the genus Drosophila were selectively killed by feeding on short (<40 nt) dsRNAs that targeted the 3' UTR of the gamma-tubulin gene. For the aphid nymphs and beetle and moth larvae, dsRNA could simply be dissolved into their diets, but to induce RNAi in the drosophilid species, the dsRNAs needed to be encapsulated in liposomes to help facilitate uptake of the dsRNA. This is the first demonstration of RNAi following ingestion of dsRNA in all of the species tested, and the method offers promise of both higher throughput RNAi screens and the development of a new generation of species-specific insecticides. PMID- 19815068 TI - Sequence and function of lysosomal and digestive cathepsin D-like proteinases of Musca domestica midgut. AB - Musca domestica larvae display in anterior and middle midgut contents, a proteolytic activity with pH optimum of 3.0-3.5 and kinetic properties like cathepsin D. Three cDNAs coding for preprocathepsin D-like proteinases (ppCAD 1, ppCAD 2, ppCAD 3) were cloned from a M. domestica midgut cDNA library. The coded protein sequences included the signal peptide, propeptide and mature enzyme that has all conserved catalytic and substrate binding residues found in bovine lysosomal cathepsin D. Nevertheless, ppCAD 2 and ppCAD 3 lack the characteristic proline loop and glycosylation sites. A comparison among the sequences of cathepsin D-like enzymes from some vertebrates and those found in M. domestica and in the genomes of Aedes aegypti, Drosophila melanogaster, Tribolium castaneum, and Bombyx mori showed that only flies have enzymes lacking the proline loop (as defined by the motif: DxPxPx(G/A)P), thus resembling vertebrate pepsin. ppCAD 3 should correspond to the digestive cathepsin D-like proteinase (CAD) found in enzyme assays because: (1) it seems to be the most expressed CAD, based on the frequency of ESTs found. (2) The mRNA for CAD 3 is expressed only in the anterior and proximal middle midgut. (3) Recombinant procathepsin D-like proteinase (pCAD 3), after auto-activation has a pH optimum of 2.5-3.0 that is close to the luminal pH of M. domestica midgut. (4) Immunoblots of proteins from different tissues revealed with anti-pCAD 3 serum were positive only in samples of anterior and middle midgut tissue and contents. (5) CAD 3 is localized with immunogold inside secretory vesicles and around microvilli in anterior and middle midgut cells. The data support the view that on adapting to deal with a bacteria rich food in an acid midgut region, M. domestica digestive CAD resulted from the same archetypical gene as the intracellular cathepsin D, paralleling what happened with vertebrates. The lack of the proline loop may be somehow associated with the extracellular role of both pepsin and digestive CAD 3. PMID- 19815069 TI - Does corazonin signal nutritional stress in insects? AB - The undecapeptide corazonin, initially discovered from the American cockroach as a strong cardioaccelerator, is now known to be ubiquitously present in arthropods, although it is absent from some species, notably Coleoptera. The structure of its precursor is similar to the GnRH precursor, while it acts through a receptor related to the GnRH receptor; corazonin thus appears to be an arthropod homolog of GnRH. It is produced by neuroendocrine cells in the brain, as well as interneurons in the ventral nerve cord. These two cell types are generally present in insects; in most species there are also other neurons producing corazonin. Its function in insects has remained obscure; its cardioacceleratory effects are limited to a few cockroach species, while in other species different physiological effects have been described. Most spectacularly it induces changes associated with the gregarious phase in migratory locusts and in the silkworm it reduces the size of the cocoon formed. Corazonin is able to induce ecdysis in two moth species, however locusts and flies in which the corazonin gene is no longer expressed, ecdyse normally and, hence, it is not clear whether corazonin is essential for ecdysis. As the corazonin neuroendocrine cells in the brain express receptors for two midgut peptides, it seems likely that their activity is modulated by the midgut endocrine cells. I propose that in insects corazonin might be released under conditions of nutritional stress, which can explain several of the observed physiological effects of this neurohormone. PMID- 19815070 TI - Induction of chronic renal allograft dysfunction in a rat model with complete and exclusive MHC incompatibility. AB - Chronic allograft dysfunction is one of the most important reasons for late graft loss after renal transplantation. Its etiology is multifactorial and combines immunological as well as non-immunological mechanisms. It is known from large registry data that MHC mismatches are inversely correlated to long term allograft survival. Although this is a well known aspect, the mechanisms of MHC-driven graft damage and the impact of other immunological factors leading to chronic rejection are poorly understood. In patients it is impossible to study MHC mismatches without considering non-MHC differences. Further more common animal models for chronic rejection are all characterized by non-MHC as well as MHC disparities. To exclusively study MHC mediated immunoresponses we established a rat model of renal transplantation using congenic rat strains differing in their entire MHC class I and class II, but sharing the genetic background of the LEW rat. After an initial short term of immunosuppression all animals developed renal impairment with severe albuminuria. Half of the animals died of renal failure in week 7 to 14 and showed pathological characteristics of chronic allograft damage including IF/TA and severe glomerulopathy. The majority of these recipients developed circulating donor-specific MHC alloantibodies. Allografts were significantly infiltrated with T-cells, macrophages and NK-cells. We established a MHC congenic rat model to investigate immunological mechanisms of chronic renal allograft rejection exclusively induced by a complete MHC mismatch. We demonstrated humoral as well as cellular immunoresponses leading to chronic allograft loss in 50% of animals. PMID- 19815071 TI - An endocannabinoid tone limits excitotoxicity in vitro and in a model of multiple sclerosis. AB - The aim of this study was to evaluate how endocannabinoids interact with excitotoxic processes both in vitro, using primary neural cell cultures, and in vivo, in the TMEV-IDD model of multiple sclerosis. First, we observed that neuronal cells respond to excitotoxic challenges by the production of endocannabinoid molecules which in turn exerted neuroprotective effects against excitotoxicity. The inhibitor of endocannabinoid uptake, UCM707, protected specifically against AMPA-induced excitotoxicity, by activating CB(1) and CB(2) cannabinoid receptors, as well as the nuclear factor, PPARgamma. This neuroprotective effect was reverted by blocking the glial glutamate transporter, GLT-1. Mice subjected to the model of multiple sclerosis showed a decrease in the expression of GLT-1. UCM707 reversed this loss of GLT-1 and induced a therapeutic effect. Our data indicate that the enhancement of the endocannabinoid tone leads to neuroprotection against AMPA-induced excitotoxicity and provides therapeutic effects in this model of multiple sclerosis. PMID- 19815072 TI - Interactions of peptide amidation and copper: novel biomarkers and mechanisms of neural dysfunction. AB - Mammalian genomes encode only a small number of cuproenzymes. The many genes involved in coordinating copper uptake, distribution, storage and efflux make gene/nutrient interactions especially important for these cuproenzymes. Copper deficiency and copper excess both disrupt neural function. Using mice heterozygous for peptidylglycine alpha-amidating monooxygenase (PAM), a cuproenzyme essential for the synthesis of many neuropeptides, we identified alterations in anxiety-like behavior, thermoregulation and seizure sensitivity. Dietary copper supplementation reversed a subset of these deficits. Wildtype mice maintained on a marginally copper-deficient diet exhibited some of the same deficits observed in PAM(+/-) mice and displayed alterations in PAM metabolism. Altered copper homeostasis in PAM(+/-) mice suggested a role for PAM in the cell type specific regulation of copper metabolism. Physiological functions sensitive to genetic limitations of PAM that are reversed by supplemental copper and mimicked by copper deficiency may serve as indicators of marginal copper deficiency. PMID- 19815073 TI - Overproduction, purification and characterisation of Tbj1, a novel Type III Hsp40 from Trypanosoma brucei, the African sleeping sickness parasite. AB - The heat shock protein 40 (Hsp40) family of proteins act as co-chaperones of the heat shock protein 70 (Hsp70) chaperone family, and together they play a vital role in the maintenance of cellular homeostasis. The Type III class of Hsp40s are diverse in terms of both sequence identity and function and have not been extensively characterised. The Trypanosoma brucei parasite is the causative agent of Human African Trypanosomiasis, and possesses an unusually large Hsp40 complement, consisting mostly of Type III Hsp40s. A novel T. brucei Type III Hsp40, Tbj1, was heterologously expressed, purified, and found to exist as a compact monomer in solution. Using polyclonal antibodies to the full-length recombinant protein, Tbj1 was found by Western analysis to be expressed in the T. brucei bloodstream-form. Tbj1 was found to be able to assist two different Hsp70 proteins in the suppression of protein aggregation in vitro, despite being unable to stimulate their ATPase activity. This indicated that while Tbj1 did not possess independent chaperone activity, it potentially functioned as a novel co chaperone of Hsp70 in T. brucei. PMID- 19815074 TI - Purification of RNA polymerase from mycobacteria for optimized promoter polymerase interactions. AB - In vitro transcription analysis is important to understand the mechanism of transcription. Various assays for the analysis of initiation, elongation and termination form the basis for better understanding of the process. Purified RNA polymerase (RNAP) with high specific activity is necessary to carry out variety of these specific reactions. The RNAP purified from Mycobacterium smegmatis from exponential phase showed low promoter specificity in promoter-polymerase interaction studies. This is due to the presence of a large number of sigma factors during exponential phase and under-representation of sigma(A) required for house-keeping transcription. We describe an in vivo reconstitution of RNAP holoenzyme with sigma(A) and its purification, which resulted in holoenzyme with stoichiometric sigma(A) content. The reconstituted holoenzyme showed enhanced promoter-specific binding and promoter-specific-transcription activity compared to the enzyme isolated using standard procedure. Such in vivo reconstitution of stoichiometric holoenzyme could facilitate promoter-specific transcription assays, especially in organisms which encode a large number of sigma factors. PMID- 19815075 TI - Use of a new catabolite repression resistant promoter isolated from Bacillus subtilis KCC103 for hyper-production of recombinant enzymes. AB - Bacillus subtilis KCC103 hyper-produces alpha-amylase and the synthesis is resistant to carbon catabolite repression. The strain efficiently produced alpha amylase in low cost agro-biomass based medium rich in simple sugars without catabolite repression. Here, the catabolite repression resistant promoter (amyR4) of alpha-amylase was isolated from KCC103 and used to synthesize recombinant enzymes in B. subtilis. When the bgaB gene encoding beta-galactosidase of Bacillus stearothermophilus was cloned and expressed under the amyR4 promoter, high level of beta-galactosidase activity was found in Escherichia coli (28 U/ml)) and B. subtilis (19 U/ml). Further, the genes encoding endoxylanase (xynA) and carboxymethyl cellulase (bglC) from B. subtilis were cloned with signal peptides and expressed with CCR-resistant amyR4 promoter. In E. coli, the expression was intracellular with activities of cellulase and xylanase at 76 and 105IU/ml respectively. The expression was extracellular in B. subtilis with activities at 17 and 17 IU/ml of cellulase and xylanase respectively in LB medium. When recombinant B. subtilis was cultured in LB-glucose medium, the synthesis of recombinant enzymes was not subject to catabolite repression and the expression was observed throughout the growth. This is important as glucose in the medium can prevent sporulation of the Bacillus and prevent activation of the other scavenger pathways that leads to degradation of recombinant proteins. The catabolite derepressed promoter of alpha-amylase from B. subtilis KCC103 can be efficiently used for overexpression of various industrial enzymes. PMID- 19815076 TI - Screening of effective column rinse solvent for Protein-A chromatography. AB - Mildly acidic arginine solution is highly effective in elution of bound proteins from Protein-A columns. Although Protein-A is specific in antibody capture, it does bind other proteins, which must then be removed before elution by aqueous arginine solution. If they are not removed, a strong elution property of aqueous arginine solutions will elute the contaminating proteins along with antibodies. Here we have examined various salt solutions as a column rinse solvent. We screened various solvents for their effects on binding of purified antibodies to Protein-A, instead of their effectiveness to elute the bound contaminants. Those solvents that result in a slight flow-through of the antibodies during loading should be effective in eluting non-specifically bound proteins that have weaker affinity for Protein-A than antibodies: namely, if a particular solvent reduces antibody binding to Protein-A, it is expected to be effective in reducing binding of contaminants and hence eluting them. Such screening showed a few compounds, including arginine and sodium acetate, as potential column rinse agents. A combination of arginine and sodium acetate was tested for a few crude materials containing antibodies. PMID- 19815077 TI - Molecular cloning and characterization of a heat shock protein 70 gene in swimming crab (Portunus trituberculatus). AB - Hsp70 can stimulate cells of the innate immune system directly by acting as "danger"-signaling molecules. To understand the immune defense mechanisms of swimming crab Portunus trituberculatus (Decapoda: Brachyura: Portunidae), the cDNA of Hsp70 (designated PtHsp70) was cloned by the combination of homology cloning and rapid amplification of cDNA ends (RACE). The full-length PtHsp70 cDNA was 2195 bp, including an open reading frame (ORF) of 1950 bp encoding a polypeptide of 650 amino acids with estimated molecular mass of 71.1514 kDa and theoretical isoelectric point of 5.38. FASTA and BLAST analysis indicated that PtHsp70 should be an inducible cytosolic member of the Hsp70 family. The coding region of PtHsp70 was uninterrupted and four SNPs with 1133C/T, 1311C/T, 1551C/T and 1809 A/G were detected by direct sequencing of 20 genomic samples. Using fluorescent real-time quantitative PCR, the transcriptional expression of PtHsp70 showed a clear time-dependent response after challenge by Vibrio alginolyticus, the main causative agent of emulsification disease causing large mortality in P. trituberculatus. This is the first report on the expression of Hsp70 induced by pathogen stimulation in Brachyura. Phylogenetic analysis revealed that the inducible Hsp70s were divided into two groups in crab and PtHsp70 was clustered into the Hsp/Hsc group (Clade I) by maximum-likelihood (ML) and Bayesian inference (BI) methods. GAP repeat and GGMP motif of inducible Hsp70 gene in the crab species were only found in Clade I. PMID- 19815078 TI - A new methodology for the estimation of fiber populations in the white matter of the brain with the Funk-Radon transform. AB - The Funk-Radon Transform (FRT) is a powerful tool for the estimation of fiber populations with High Angular Resolution Diffusion Imaging (HARDI). It is used in Q-Ball imaging (QBI), and other HARDI techniques such as the recent Orientation Probability Density Transform (OPDT), to estimate fiber populations with very few restrictions on the diffusion model. The FRT consists in the integration of the attenuation signal, sampled by the MRI scanner on the unit sphere, along equators orthogonal to the directions of interest. It is easily proved that this calculation is equivalent to the integration of the diffusion propagator along such directions, although a characteristic blurring with a Bessel kernel is introduced. Under a different point of view, the FRT can be seen as an efficient way to compute the angular part of the integral of the attenuation signal in the plane orthogonal to each direction of the diffusion propagator. In this paper, Stoke's theorem is used to prove that the FRT can in fact be used to compute accurate estimates of the true integrals defining the functions of interest in HARDI, keeping the diffusion model as little restrictive as possible. Varying the assumptions on the attenuation signal, we derive new estimators of fiber orientations, generalizing both Q-Balls and the OPDT. Extensive experiments with both synthetic and real data have been intended to show that the new techniques improve existing ones in many situations. PMID- 19815079 TI - Learning partially directed functional networks from meta-analysis imaging data. AB - We propose a new exploratory method for the discovery of partially directed functional networks from fMRI meta-analysis data. The method performs structure learning of Bayesian networks in search of directed probabilistic dependencies between brain regions. Learning is based on the co-activation of brain regions observed across several independent imaging experiments. In a series of simulations, we first demonstrate the reliability of the method. We then present the application of our approach in an extensive meta-analysis including several thousand activation coordinates from more than 500 imaging studies. Results show that our method is able to automatically infer Bayesian networks that capture both directed and undirected probabilistic dependencies between a number of brain regions, including regions that are frequently observed in motor-related and cognitive control tasks. PMID- 19815080 TI - LEAP: learning embeddings for atlas propagation. AB - We propose a novel framework for the automatic propagation of a set of manually labeled brain atlases to a diverse set of images of a population of subjects. A manifold is learned from a coordinate system embedding that allows the identification of neighborhoods which contain images that are similar based on a chosen criterion. Within the new coordinate system, the initial set of atlases is propagated to all images through a succession of multi-atlas segmentation steps. This breaks the problem of registering images that are very "dissimilar" down into a problem of registering a series of images that are "similar". At the same time, it allows the potentially large deformation between the images to be modeled as a sequence of several smaller deformations. We applied the proposed method to an exemplar region centered around the hippocampus from a set of 30 atlases based on images from young healthy subjects and a dataset of 796 images from elderly dementia patients and age-matched controls enrolled in the Alzheimer's Disease Neuroimaging Initiative (ADNI). We demonstrate an increasing gain in accuracy of the new method, compared to standard multi-atlas segmentation, with increasing distance between the target image and the initial set of atlases in the coordinate embedding, i.e., with a greater difference between atlas and image. For the segmentation of the hippocampus on 182 images for which a manual segmentation is available, we achieved an average overlap (Dice coefficient) of 0.85 with the manual reference. PMID- 19815082 TI - Static images of novel, moveable objects learned through touch activate visual area hMT+. AB - Although many studies have found similar cortical areas activated during the recognition of objects encoded through vision or touch, little is known about cortical areas involved in the crossmodal recognition of dynamic objects. Here, we investigated which cortical areas are involved in the recognition of moving objects and were specifically interested in whether motion areas are involved in the recognition of dynamic objects within and across sensory modalities. Prior to scanning, participants first learned to recognise a set of 12 novel objects, each presented either visually or haptically, and either moving or stationary. We then conducted fMRI whilst participants performed an old-new task with static images of learned or not-learned objects. We found the fusiform and right inferior frontal gyri more activated to within-modal visual than crossmodal object recognition. Our results also revealed increased activation in area hMT+, LOC and the middle occipital gyrus, in the right hemisphere only, for the objects learned as moving compared to the learned static objects, regardless of modality. We propose that the network of cortical areas involved in the recognition of dynamic objects is largely independent of modality and have important implications for understanding the neural substrates of multisensory dynamic object recognition. PMID- 19815083 TI - Diffusion orientation transform revisited. AB - Diffusion orientation transform (DOT) is a powerful imaging technique that allows the reconstruction of the microgeometry of fibrous tissues based on diffusion MRI data. The three main error sources involving this methodology are the finite sampling of the q-space, the practical truncation of the series of spherical harmonics and the use of a mono-exponential model for the attenuation of the measured signal. In this work, a detailed mathematical description that provides an extension to the DOT methodology is presented. In particular, the limitations implied by the use of measurements with a finite support in q-space are investigated and clarified as well as the impact of the harmonic series truncation. Near- and far-field analytical patterns for the diffusion propagator are examined. The near-field pattern makes available the direct computation of the probability of return to the origin. The far-field pattern allows probing the limitations of the mono-exponential model, which suggests the existence of a limit of validity for DOT. In the regimen from moderate to large displacement lengths the isosurfaces of the diffusion propagator reveal aberrations in form of artifactual peaks. Finally, the major contribution of this work is the derivation of analytical equations that facilitate the accurate reconstruction of some orientational distribution functions (ODFs) and skewness ODFs that are relatively immune to these artifacts. The new formalism was tested using synthetic and real data from a phantom of intersecting capillaries. The results support the hypothesis that the revisited DOT methodology could enhance the estimation of the microgeometry of fiber tissues. PMID- 19815081 TI - A comparison of fMRI adaptation and multivariate pattern classification analysis in visual cortex. AB - Functional magnetic resonance imaging (fMRI) has become a ubiquitous tool in cognitive neuroscience. The technique allows noninvasive measurements of cortical responses in the human brain, but only on the millimeter scale. Because a typical voxel contains many thousands of neurons with varied properties, establishing the selectivity of their responses directly is impossible. In recent years, two methods using fMRI aimed at studying the selectivity of neuronal populations on a 'subvoxel' scale have been heavily used. The first technique, fMRI adaptation, relies on the observation that the blood oxygen level-dependent (BOLD) response in a given voxel is reduced after prolonged presentation of a stimulus, and that this reduction is selective to the characteristics of the repeated stimuli (adapters). The second technique, multivariate pattern analysis (MVPA), makes use of multivariate statistics to recover small biases in individual voxels in their responses to different stimuli. It is thought that these biases arise due to the uneven distribution of neurons (with different properties) sampled by the many voxels in the imaged volume. These two techniques have not been compared explicitly, however, and little is known about their relative sensitivities. Here, we compared fMRI results from orientation-specific visual adaptation and orientation-classification by MVPA, using optimized experimental designs for each, and found that the multivariate pattern classification approach was more sensitive to small differences in stimulus orientation than the adaptation paradigm. Estimates of orientation selectivity obtained with the two methods were, however, very highly correlated across visual areas. PMID- 19815084 TI - X inactivation and disease. AB - X inactivation is the mechanism by which mammals adjust the X-linked gene dosage between the sexes. The dosage difference between XX females and XY males is functionally equalized by silencing one of the two X chromosomes in female cells. This dosage-compensation mechanism is based on the long functional Xist RNA. Here, we review our understanding of dosage compensation and Xist function in the context of disease. PMID- 19815085 TI - Restoring chromatin after replication: how new and old histone marks come together. AB - In dividing cells genome stability and function rely on faithful transmission of both DNA sequence and its organization into chromatin. In the course of DNA replication chromatin undergoes transient genome-wide disruption followed by restoration on new DNA. This involves tight coordination of DNA replication and chromatin assembly processes in time and space. Dynamic recycling and de novo deposition of histones are fundamental for chromatin restoration. Histone post translational modifications (PTMs) are thought to have a causal role in establishing distinct chromatin structures. Here we discuss PTMs present on new and parental histones and how they influence genome stability and restoration of epigenetically defined domains. Newly deposited histones must change their signature in the process of chromatin restoration, this may occur in a step-wise fashion involving replication-coupled processes and information from recycled parental histones. PMID- 19815086 TI - Planar cell polarity and cilia. AB - In the last few years, evidence has come to light suggesting that planar cell polarity signaling in vertebrates may be controlled and modulated by primary cilia, subcellular organelles that emerge from the plasma membrane of most cell types. This characteristic distinguishes vertebrate planar cell polarity signaling from that in insects. We review here some of the experimental evidence contributing to this finding. These observations have begun to suggest molecular and cellular mechanisms of the so-called ciliopathies, important human diseases characterized by defective ciliary functions. PMID- 19815087 TI - Chromatin regulation and non-coding RNAs at mammalian telomeres. AB - In eukaryotes, terminal chromosome repeats are bound by a specialized nucleoprotein complex that controls telomere length and protects chromosome ends from DNA repair and degradation. In mammals the "shelterin" complex mediates these central functions at telomeres. In the recent years it has become evident that also the heterochromatic structure of mammalian telomeres is implicated in telomere length regulation. Impaired telomeric chromatin compaction results in a loss of telomere length control. Progressive telomere shortening affects chromatin compaction at telomeric and subtelomeric repeats and activates alternative telomere maintenance mechanisms. Dynamics of chromatin structure of telomeres during early mammalian development and nuclear reprogramming further indicates a central role of telomeric heterochromatin in organismal development. In addition, the recent discovery that telomeres are transcribed, giving rise to UUAGGG-repeat containing TelRNAs/TERRA, opens a new level of chromatin regulation at telomeres. Understanding the links between the epigenetic status of telomeres, TERRA/TelRNA and telomere homeostasis will open new avenues for our understanding of organismal development, cancer and ageing. PMID- 19815088 TI - Viral vectors for the treatment of alcoholism: use of metabolic flux analysis for cell cultivation and vector production. AB - The HEK293 cell line has been used for the production of adenovirus vectors to be used in the potential treatment of alcoholism using a gene therapy strategy. Culture optimization and scale-up has been achieved by first adapting the cells to serum-free media and secondly by growing them in suspension. Adenovirus production after infection was increased, resulting in higher specific glucose consumption and lactate accumulation rates compared to the growth phase. We applied media design tools and Metabolic Flux Analysis (MFA) to compare the metabolic states of cells during growth and adenovirus production and to optimize culture media according to the metabolic demand of the cells in terms of glucose and glutamine concentrations. This allowed obtaining a higher maximum cell concentration and increased adenovirus production by minimizing the production of metabolites that can have an inhibitory effect on cell growth. We have proposed a stoichiometric equation for adenovirus synthesis. MFA results allowed determination of how these changes in composition affected the way cells distribute their nutrient resources during cell growth and virus production. Virus purification was successfully achieved using chromatography and Aqueous Two Phase Systems (ATPS). PMID- 19815089 TI - Drug discovery dilemma and Cura quartet collaboration. AB - Many parties contribute to discovery of new drugs - academic researchers, industry scientists, government agencies, and disease foundation helping to corral the resources necessary to sustain research efforts - but it has never been more apparent until now that these parties must work together to accomplish the shared goal of improving health. At a recent conference at the Georgetown University Medical Center, a group of prominent scientists from the academic, industry, government and disease advocacy communities came together to discuss new paths forward for stronger inter-institutional collaboration to establish a framework for translating new discoveries into drugs, improving proof of concept (PoC) studies, and reducing attrition at the clinical stage of drug development. PMID- 19815090 TI - Do DNA repair genes OGG1, XRCC3 and XRCC7 have an impact on susceptibility to bladder cancer in the North Indian population? AB - OBJECTIVE: Polymorphisms in DNA repair genes may be associated with altered DNA repair capacity, thereby influencing an individual's susceptibility to smoking related cancers such as bladder cancer. Therefore, we sought to examine the correlation between single nucleotide polymorphisms in DNA repair genes and bladder cancer. METHODOLOGY: We undertook a case-control study of 212 urothelial bladder cancer (UBC) cases and 250 controls to investigate the association between OGG1 (C1245G rs1052133), XRCC3 (C18067T, rs861539) and XRCC7 (G6721T, rs7003908) polymorphisms and bladder cancer susceptibility by PCR-RFLP and the ARMS method. We also investigated gene-environment interactions. RESULTS: The OGG1 GG genotype was associated with an elevated risk of urothelial bladder cancer (UBC) (OR, 2.10; p, 0.028). XRCC7 + 6721 GG was also associated with increased susceptibility to UBC (OR, 4.45; p, 0.001). In a recessive model, the OGG1 GG genotype showed an increased risk of TaG(2,3) + T1G(1-3) tumors. Additionally, the OGG1 GG genotype in non-smokers represented a 2.46-fold greater risk (OR, 2.46; p, 0.035) in bladder cancer patients. Subsequent analysis demonstrated more pronounced association of XRCC7 with smokers (OR, 4.39; p, 0.001). XRCC7 also showed increased association with TaG(2,3) + T1G(1-3) tumors and muscle invasive tumors (OR, 3.16; p, 0.001 and OR, 4.24; p, 0.001, respectively). Multiple Cox regression analysis in non-muscle invasive bladder tumor (NMIBT) patients demonstrated an association of the OGG1 GG polymorphism with a high risk of recurrence in patients on cystoscopic surveillance (HR, 4.04; p, 0.013). Subsequently, shorter recurrence-free survival (log rank p, 0.024; CC/GG, 42/24) was observed. CONCLUSION: Our data suggest association of a variant (GG) genotype of OGG1 with increased UBC susceptibility and a high risk of tumor recurrence in NMIBT patients on cystoscopic surveillance. XRCC7 G allele carriers (TG+GG) are also at an elevated risk for susceptibility to UBC as evidenced by a high odds ratio throughout the analysis. PMID- 19815091 TI - Thymol, a naturally occurring monocyclic dietary phenolic compound protects Chinese hamster lung fibroblasts from radiation-induced cytotoxicity. AB - The effect of thymol (TOH), a dietary compound was investigated for its ability to protect against radiation-induced cytotoxicity in Chinese hamster lung fibroblast (V79) cells growing in vitro. Treatment of V79 cells with 25 microg/ml of TOH prior to 10 Gy gamma radiation resulted increase in the cell viability than that of radiation alone as evaluated by MTT assay. Similarly, there was a significant increase in the surviving fraction observed with 25 microg/ml of TOH administered 1h prior to graded doses of gamma radiation. Further, 25 microg/ml TOH treatment before irradiation significantly decreased the percentage of radiation-induced apoptotic cells (sub-G(1) population) analyzed by flow cytometry as well as DNA ladder assay. TOH was found to inhibit various free radicals generated in vitro, viz., DPPH, O(2), ABTS(+) and OH in a concentration dependent manner. TOH also inhibited the radiation-induced decrease in intracellular glutathione, superoxide dismutase and catalase enzyme levels in V79 cells accompanied by the reduction in lipid peroxides. Our study demonstrated antagonistic potential of TOH against radiation-induced oxidative stress, lipid peroxidation resulting in increased cell viability. PMID- 19815092 TI - Analysis of charge and mass effects on peroxidase expressions and activities in Arabidopsis thaliana after low-energy ion irradiation. AB - In order to study the impact of incident ions on organism mutation, positively and negatively charged low-energy oxygen and hydrogen ions were implanted into the dry seeds of Arabidopsis thaliana using three different dosages. The effects of ion irradiation on peroxidase abundance, activity, isozyme patterns, and transcription were analyzed. The results showed that ion irradiation increases peroxidase activity, transcription, and translation. In all analyses, the relative magnitude of effect of the three ions was consistent, with the implantation of O(+) and H(+) affecting peroxidase more than O(-). This finding suggests that the charge of an incident ion influences cellular systems more than its mass. PMID- 19815093 TI - NDRG2 induced by oxidized LDL in macrophages antagonizes growth factor productions via selectively inhibiting ERK activation. AB - During atherogenesis, macrophage foam cells produce prodigious growth factors, cytokines, and chemokines, which play the central roles in inflammatory process in atherosclerotic plaque formation. In the present study, we identified a new protein marker, N-Myc downstream-regulated protein 2 (NDRG2), which is significantly up-regulated in oxidized low density lipoprotein (oxLDL) treated macrophages and in human atherosclerotic plaques. Over-expression and siRNA knockdown studies showed that NDRG2 is a negative regulator of platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) productions in macrophages. Furthermore, we investigated the effects of NDRG2 on MAPK signal activation. Our results showed ERK1/2 activation, but not P38 or JNK1/2 activation, is responsible for regulation of NDRG2 on VEGF and PDGF productions. Consistent with the PDGF levels, the vascular smooth muscle cell (VSMC) proliferation was also regulated by the conditional medium of the oxLDL treated macrophages with NDRG2 knockdown or over-expression. Neutralizing anti-PDGF antibody can significantly inhibit the enhanced VSMC proliferation by macrophage medium with NDRG2 knockdown. Our present results demonstrate that NDRG2 participates in oxLDL-induced macrophage activation and modulates ERK1/2 dependent PDGF and VEGF production, which has potential application in atherogenesis. PMID- 19815094 TI - Alendronate affects calcium dynamics in cardiomyocytes in vitro. AB - Therapy with bisphosphonates, including alendronate (ALN), is considered a safe and effective treatment for osteoporosis. However, recent studies have reported an unexpected increase in serious atrial fibrillation (AF) in patients treated with bisphosphonates. The mechanism that explains this side effect remains unknown. Since AF is associated with an altered sarcoendoplasmic reticulum calcium load, we studied how ALN affects cardiomyocyte calcium homeostasis and protein isoprenylation in vitro. Acute and long-term (48h) treatment of atrial and ventricular cardiomyocytes with ALN (10(-8)-10(-6)M) was performed. Changes in calcium dynamics were determined by both fluorescence measurement of cytosolic free Ca(2+) concentration and western blot analysis of calcium-regulating proteins. Finally, effect of ALN on protein farnesylation was also identified. In both atrial and ventricular cardiomyocytes, ALN treatment delayed and diminished calcium responses to caffeine. Only in atrial cells, long-term exposure to ALN induced transitory calcium oscillations and led to the development of oscillatory component in calcium responses to caffeine. Changes in calcium dynamics were accompanied by changes in expression of proteins controlling sarcoendoplasmic reticulum calcium. In contrast, ALN minimally affected protein isoprenylation in these cells. In summary, treatment of atrial cardiomyocytes with ALN-induced abnormalities in calcium dynamics consistent with induction of a self stimulatory, pacemaker-like behavior, which may contribute to the development of cardiac side effects associated with these drugs. PMID- 19815095 TI - Aortoenteric fistula: an uncommon cause of hematemesis with characteristic clinical presentation. PMID- 19815096 TI - Effect of process control agent on the porous structure and mechanical properties of a biomedical Ti-Sn-Nb alloy produced by powder metallurgy. AB - The influence of different amounts and types of process control agent (PCA), i.e., stearic acid and ethylene bis-stearamide, on the porous structure and mechanical properties of a biomedical Ti-16Sn-4Nb (wt.%) alloy was investigated. Alloy synthesis was performed on elemental metal powders using high-energy ball milling for 5h. Results indicated that varying the PCA content during ball milling led to a drastic change in morphology and particle-size distribution of the ball-milled powders. Porous titanium alloy samples sintered from the powders ball milled with the addition of various amounts of PCA also revealed different pore morphology and porosity. The Vickers hardness of the sintered titanium alloy samples exhibited a considerable increase with increasing PCA content. Moreover, the addition of larger amounts of PCA in the powder mixture resulted in a significant increase in the elastic modulus and peak stress for the sintered porous titanium alloy samples under compression. It should also be mentioned that the addition of PCA introduced contamination (mainly carbon and oxygen) into the sintered porous product. PMID- 19815097 TI - Developments in metallic biodegradable stents. AB - Interest in metallic degradable biomaterials research has been growing in the last decade. Both scientific journals and patent databases record a high increase in publications in this area. Biomedical implants with temporary function, such as coronary stents, are the targeted applications for this novel class of biomaterials. It is expected that stents made of degradable biomaterials, named biodegradable stents, will provide a temporary opening into a narrowed arterial vessel until the vessel remodels and will progressively disappear thereafter. Biodegradable stents made of metal have recently been progressed into preclinical tests in humans after their first introduction in early 2000s. By referring to patents and journal publications, this paper reviews the developments in biodegradable stents, with emphasis on those made of metals, starting from the first design ideas to validation testing. PMID- 19815098 TI - In vitro degradation and cytotoxicity of Mg/Ca composites produced by powder metallurgy. AB - Mg/Ca (1 wt.%, 5 wt.%, 10 wt.% Ca) composites were prepared from pure magnesium and calcium powders using the powder metallurgy method, aiming to enlarge the addition of Ca content without the formation of Mg(2)Ca. The microstructures, mechanical properties and cytotoxicities of Mg/Ca composite samples were investigated. The corrosion of Mg/Ca composites in Dulbecco's modified Eagle's medium (DMEM) for various immersion intervals was studied by electrochemical impedance spectroscopy measurements and environmental scanning electron microscope, with the concentrations of released Mg and Ca ions in DMEM for various immersion time intervals being measured. It was shown that the main constitutional phases were Mg and Ca, which were uniformly distributed in the Mg matrix. The ultimate tensile strength (UTS) and elongation of experimental composites decreased with increasing Ca content, and the UTS of Mg/1Ca composite was comparable with that of as-extruded Mg-1Ca alloy. The corrosion potential increased with increasing Ca content, whereas the current density and the impedance decreased. It was found that the protective surface film formed quickly at the initial immersion stage. With increasing immersion time, the surface film became compact, and the corrosion rate of Mg/Ca composites slowed down. The surface film consisted mainly of CaCO(3), MgCO(3)x3H(2)O, HA and Mg(OH)(2) after 72 h immersion in DMEM. Mg/1Ca and Mg/5Ca composite extracts had no significant toxicity (p>0.05) to L-929 cells, whereas Mg/10Ca composite extract induced approximately 40% reduced cell viability. PMID- 19815099 TI - On the in vitro and in vivo degradation performance and biological response of new biodegradable Mg-Y-Zn alloys. AB - A design strategy deployed in developing new biodegradable Mg-Y-Zn alloys is summarized and the key factors influencing their suitability for medical applications are described. The Mg-Y-Zn alloys reveal microstructural features and mechanical characteristics expected to be appropriate for vascular intervention applications. The focus of this article lies in the evaluation of the degradation performance and biological response of the alloys with respect to their potential as implant materials (stents). The degradation characteristics analyzed by immersion testing and electrochemical impedance spectroscopy in simulated physiological media reveal slow and homogeneous degradation. In vitro cell tests using human umbilical vein endothelial cells indicate good cytocompatibility on the basis of the alloys' eluates (extracts). Animal studies carried out with pigs on Mg-2Y-1Zn (in wt.%) reveal an auspicious in vivo performance. Evaluation of preparations derived from implants in various types of tissues indicates homogeneous degradation and only limited gas formation during in vivo testing. The characteristics of the tissue reactions indicate good biocompatibility. The new Mg-Y-Zn alloys show an interesting combination of preferred microstructural, mechanical, electrochemical and biological properties, which make them very promising for degradable implant applications. PMID- 19815100 TI - Evaluation of composition and crosslinking effects on collagen-based composite constructs. AB - Vascular grafts are widely used for a number of medical treatments. Strength, compliance, endothelialization and availability are issues of most concern for vascular graft materials. With current approaches, these requirements are difficult to satisfy simultaneously. To explore an alternative approach, the present study has engineered the collagen gel construct by incorporating mimetic components and crosslinking the construct with different crosslinkers. The effects of component additives, such as chitosan and elastin, have been evaluated in terms of their mechanical and biological properties. Results demonstrate that the incorporation of chitosan and/or elastin alter stress-strain curves in the low stress loading region, and significantly improve the stretching ratio and ultimate stress of gel constructs compared to collagen constructs. Electron microscopy results suggest that the mechanical improvements might be due to microstructural modifications by chitosan sheets and elastin fibers. The effects of crosslinkers, such as formaldehyde, genipin and ethyl-(dimethyl aminopropyl) and carbodiimide hydrochloride (EDAC) have also been evaluated. Results demonstrate that formaldehyde, EDAC and genipin employ different mechanisms to crosslink collagen-based constructs, and use of genipin as a construct crosslinker exhibits improved elongation and endothelial coverage as compared to formaldehyde and EDAC. In addition, extending gelation time increased the elastic modulus but not the ultimate strength. Therefore, this study suggests that the mimicry of natural vessel tissues with properly crosslinked biopolymer composites could be a potential material design strategy for vascular graft materials. PMID- 19815101 TI - In vivo performance of absorbable collagen sponges with rosuvastatin in critical size cortical bone defects. AB - Rosuvastatin (RSV) is a synthetic statin with favourable pharmacologic properties, but its local effect in bone has yet to be investigated. The aim of this study was to evaluate the potential of absorbable collagen sponge (ACS) as a carrier for RSV to enhance bone formation in critical-size cortical bone defects adjacent to titanium implants. ACS, treated with different concentrations of RSV (R1 = 8.7 + or - 1.8 microg; R2 = 52.0 + or - 4.4 microg; R3 = 259.1 + or - 8.8 microg) or phosphate-buffered saline alone, were placed into the bone marrow through a defect made in the proximal tibial cortical bone of New Zealand White rabbits. One empty defect (SHAM) served as an internal control in each animal. After a healing time of 4 weeks, a concentration-dependent increase of alkaline phosphatase activity in ACS treated with RSV was detected in the bone fluid after removing the implants. In addition, a significant concentration-dependent increase in BMP-2 mRNA levels was found in the cortical bone tissue adjacent to the RSV-treated ACS. The cortical architecture of bone defects analysed by micro computed tomography showed a trend towards higher bone volume in the ACS+R1 group compared with SHAM, which was accompanied by an increase in the bone mineral density. Evaluation of histological sections showed new bone formation in ACS treated with RSV but not in untreated ACS. These results indicate that RSV, when administered locally in bone, may have a potential effect in stimulating bone formation. PMID- 19815102 TI - Adult presentation of giant retroperitoneal cystic lymphangioma: case report. AB - This case describes the rare presentation of a retroperitoneal cystic lymphangioma in a 35 year old female patient. The lymphangioma ultimately progressed to the point of inducing clinical symptoms, thus requiring surgical removal - which was accomplished without incident. The relevant clinical pictures are included for educational value. PMID- 19815103 TI - Wheat quality related differential expressions of albumins and globulins revealed by two-dimensional difference gel electrophoresis (2-D DIGE). AB - Comparative proteomics analysis offers a new approach to identify differential proteins among different wheat genotypes and developmental stages. In this study, the non-prolamin expression profiles during grain development of two common or bread wheat cultivars (Triticum aestivum L.), Jing 411 and Sunstate, with different quality properties were analyzed using two-dimensional difference gel electrophoresis (2-D DIGE). Five grain developmental stages during the post anthesis period were sampled corresponding to the cumulative averages of daily temperatures ( degrees C: 156 degrees C, 250 degrees C, 354 degrees C, 447 degrees C and 749.5 degrees C). More than 400 differential protein spots detected at one or more of the developmental stages of the two cultivars were monitored, among which 230 proteins were identified by MS. Of the identified proteins, more than 85% were enzymes possessing different physiological functions. A total of 36 differential proteins were characterized between the two varieties, which are likely to be related to wheat quality attributes. About one quarter of the proteins identified expressed in multiple spots with different pIs and molecular masses, implying certain post-translational modifications (PTMs) of proteins such as phosphorylations and glycosylations. The results provide new insights into biochemical mechanisms for grain development and quality. PMID- 19815104 TI - Bone involvement in clusters of autoimmune diseases: just a complication? AB - Bone loss, described in individual groups of patients with Type 1 diabetes (T1D), autoimmune thyroid disease (ATD) or celiac disease (CD) is usually viewed as a complication of these diseases. There is increasing evidence that alterations in the immune system may directly affect bone mass. Clustering of autoimmune diseases in the same individual might predispose to higher risk of osteopenia due to imbalance in immune regulation. The aim of this study was to evaluate bone involvement in clusters of the most common autoimmune diseases (T1D, ATD and CD) in children. The study was performed at a tertiary care center for the care of pediatric diabetes. One-hundred-two patients with T1D alone or associated with ATD and/or CD were studied; 13 patients had cluster of three autoimmune diseases. Amplitude-dependent speed of sound (AD-SoS) was measured by phalangeal quantitative ultrasound and expressed as standard deviation score (SDS). AD-SoS SDS < -2 was considered indicative of osteopenia. Osteopenia was equally distributed among children with T1D alone (8.1%), T1D associated with ATD (7.7%) or CD (10.3%), while it was 53.8% in patients presenting with three autoimmune diseases. Poor compliance to gluten-free diet increased osteopenia to 18.8% in patients with T1D and CD and 80% in patients with three autoimmune disorders. No difference among groups was found with regard to gluco-metabolic control, calcium metabolism, thyroid function. In conclusion bone impairment in multiple autoimmune diseases might be considered not only a complication due to endocrine or nutritional mechanisms, but also a consequence of an immunoregulatory imbalance. Alterations of homeostatic mechanisms might explain an imbalance of osteoclast activity leading to osteopenia. PMID- 19815105 TI - Inhibition of methylation decreases osteoblast differentiation via a non-DNA dependent methylation mechanism. AB - S-adenosylmethionine (SAM)-dependent methylation of biological molecules including DNA and proteins is rapidly being uncovered as a critical mechanism for regulation of cellular processes. We investigated the effects of reduced SAM dependent methylation on osteoblast differentiation by using periodate oxidized adenosine (ADOX), an inhibitor of SAM-dependent methyltransferases. The capacity of this agent to modulate osteoblast differentiation was analyzed under non osteogenic control conditions and during growth factor-induced differentiation and compared with the effect of inhibition of DNA methylation by 5-Aza-2' deoxycytidine (5-Aza-CdR). Without applying specific osteogenic triggers, both ADOX and 5-Aza-CdR induced mRNA expression of the osteoblast markers Alp, Osx, and Ocn in murine C2C12 cells. Under osteogenic conditions, ADOX inhibited differentiation of both human mesenchymal stem cells and C2C12 cells. Gene expression analysis of early (Msx2, Dlx5, Runx2) and late (Alp, Osx, Ocn) osteoblast markers during bone morphogenetic protein 2-induced C2C12 osteoblast differentiation revealed that ADOX only reduced expression of the late phase Runx2 target genes. By using a Runx2-responsive luciferase reporter (6xOSE), we showed that ADOX reduced the activity of Runx2, while 5-Aza-CdR had no effect. Taken together, our data suggest that decreased SAM-dependent methyltransferase activity leads to impaired osteoblast differentiation via non-DNA-dependent methylation mechanisms and that methylation is a regulator of Runx2-controlled gene expression. PMID- 19815106 TI - What's in a cough? And how can it be prevented: report on the 1st Pneumonia Working Group Meeting, Seoul, 15 December 2007. PMID- 19815107 TI - Zinc-dependent binding between peptides derived from rainbow trout CD8alpha and LCK. AB - The cytoplasmic tail of mammalian CD8alpha binds the kinase LCK in a zinc dependent manner. In analogy with a previous study for humans (Kim et al., 2003) peptides were synthesized from rainbow trout CD8alpha and LCK. Surface plasmon resonance (SPR) analysis indicated that also in fish these molecules bind to each other in a zinc-dependent manner. PMID- 19815108 TI - Contrast enhanced computed tomography can predict the glycosaminoglycan content and biomechanical properties of articular cartilage. AB - OBJECTIVE: An early hallmark of osteoarthritis (OA) is the progressive loss of glycosaminoglycans (GAGs), the extracellular matrix (ECM) component of articular cartilage that confers it with compressive stiffness. Our aim in this work is to establish the feasibility of using Contrast Enhanced Computed Tomography (CECT) with an anionic iodinated contrast agent - Cysto Conray II - as a minimally invasive tool to measure the changes in the GAG content as well as the compressive stiffness of articular cartilage. METHODS: The GAG content of mated osteochondral plugs excised from bovine patello-femoral joints was progressively degraded using chondroitinase ABC. The mated plugs were then immersed in an anionic, tri-iodinated contrast agent, imaged using peripheral quantitative computed tomography (pQCT), subjected to an unconfined compressive stress relaxation test and the GAG content measured using 1,9-dimethylmethylene blue (DMMB) assay. Partial correlation analysis was performed to compare the variation in X-ray attenuation measured by pQCT to the variation in GAG content and in equilibrium compressive modulus. RESULTS: The X-ray attenuation of cartilage exposed to an anionic, tri-iodinated, contrast agent measured by quantitative computed tomography (QCT) accounted for 83% of the variation in GAG content (r(2)=0.83, P<0.0001) and 93% of the variation in the equilibrium compressive modulus (r(2)=0.93, P<0.0001). CONCLUSION: Using a mated osteochondral plug model to evaluate the biochemical composition and biomechanical properties of cartilage, this study demonstrates the interrelationships between X-ray attenuation, GAG content, and equilibrium compressive modulus, and that CECT can be used to monitor and quantify changes in the GAG content and biomechanical properties of articular cartilage. PMID- 19815109 TI - Anaphylaxis: Recent advances in assessment and treatment. AB - The incidence rate of anaphylaxis is increasing, particularly during the first 2 decades of life. Common triggers include foods, medications, and insect stings. Clinical diagnosis is based on a meticulous history of an exposure or event preceding characteristic symptoms and signs, sometimes but not always supported by a laboratory test such as an elevated serum total tryptase level. Physician initiated investigation of patients with anaphylaxis whose symptoms and signs are atypical sometimes leads to important insights into previously unrecognized triggers and mechanisms. In idiopathic anaphylaxis, in which no trigger can be confirmed by means of skin testing or measurement of specific IgE, the possibility of mastocytosis or a clonal mast cell disorder must be considered in addition to the possibility of a previously unrecognized trigger. Long-term risk reduction in patients with anaphylaxis focuses on optimal management of relevant comorbidities such as asthma and other respiratory diseases, cardiovascular disease, and mastocytosis or a clonal mast cell disorder; avoidance of the relevant confirmed allergen trigger; and relevant immunomodulation such as medication desensitization, venom immunotherapy, and possibly in the future, immunotherapy with food. Emergency preparedness for recurrence of anaphylaxis in community settings includes having epinephrine (adrenaline) autoinjectors available, knowing when and how to use them, and having a written, personalized anaphylaxis emergency action plan and up-to-date medical identification. Randomized controlled trials of the pharmacologic interventions used in an acute anaphylaxis episode are needed. PMID- 19815110 TI - Mechanisms of mast cell signaling in anaphylaxis. AB - The recent development of a consensus definition and proposed diagnostic criteria for anaphylaxis offers promise for research efforts and a better understanding of the epidemiology and pathogenesis of this enigmatic and life-threatening disease. This review examines basic principles and recent research advances in the mechanisms of mast cell signaling believed to underlie anaphylaxis. The unfolding complexity of mast cell signaling suggests that the system is sensitive to regulation by any of several individual signaling pathways and intermediates and that complementary pathways regulate mast cell activation by amplified signals. The signaling events underlying anaphylactic reactions have largely been identified through experiments in genetically modified mice and supported by biochemical studies of mast cells derived from these mice. These studies have revealed that signaling pathways exist to both upregulate and downregulate mast cell responses. In this review we will thus describe the key molecular players in these pathways in the context of anaphylaxis. PMID- 19815111 TI - Anaphylaxis syndromes related to a new mammalian cross-reactive carbohydrate determinant. AB - Anaphylaxis is a severe allergic reaction that can rapidly progress and occasionally be fatal. In instances in which the triggering allergen is not obvious, establishing the cause of anaphylaxis is pivotal to long-term management. Assigning cause is limited, however, by the number of known exposures associated with anaphylaxis. Therefore identification of novel causative agents can provide an important step forward in facilitating new, allergen-specific approaches to management. In contrast to the view that carbohydrate-directed IgE has minimal, if any, clinical significance, recent data suggest that IgE antibodies to carbohydrate epitopes can be an important factor in anaphylaxis that might otherwise appear to be idiopathic. Here we review the evidence relating to carbohydrates in food allergy and anaphylaxis and discuss the implications of a new mammalian cross-reactive carbohydrate determinant. PMID- 19815113 TI - Sexual dimorphism: Is it relevant to steroid resistance or asthma control? PMID- 19815114 TI - Reports of suicidality in clinical trials of montelukast. AB - BACKGROUND: In recent years, a number of drugs and drug classes have come under scrutiny by the US Food and Drug Administration regarding suicidality (including suicidal behavior and ideation). OBJECTIVE: We sought to perform 2 reviews (requested by the US Food and Drug Administration) of the number of events possibly related to suicidality reported in Merck clinical trials of montelukast. METHODS: Method 1 was a descriptive review of clinical adverse experiences (AEs) from 116 studies (double-blind and open-label, adult and pediatric, and single- and multiple-dose studies) completed as of March 2008. Summaries were constructed from investigator-reported AE terms possibly related to suicidality (completed suicide, suicide attempt, and suicidal ideation) or self-injurious behavior. Method 2 used a retrospective adjudication of investigator-reported AEs and other events listed in the study database described as possibly suicidality-related adverse events (PSRAEs) in a prespecified set of 41 double-blind, placebo controlled trials completed as of April 2008. RESULTS: No completed suicides were reported in any study. For the descriptive review, 20,131 adults and children received montelukast, 9,287 received placebo, and 8,346 received active control; AEs possibly related to suicidality were rare and were similar between the montelukast and placebo or active-control groups. For the adjudicated review across 22,433 patients, there were 730 adjudicated events. In 9,929 patients taking montelukast, 1 PSRAE was identified (classified as suicidal ideation); none were identified in 7,780 and 4,724 patients taking placebo and active control, respectively. CONCLUSIONS: Assessed by using 2 complementary methods, there were no reports of completed suicide, and reports of PSRAEs were rare in patients receiving montelukast and similar to those seen in control subjects. PMID- 19815115 TI - Assessing risk: Data from montelukast clinical trials. PMID- 19815116 TI - Analysis of behavior-related adverse experiences in clinical trials of montelukast. AB - BACKGROUND: Frequencies of behavior-related adverse experiences (BRAEs) in controlled clinical studies of leukotriene modifier drugs have not been summarized. OBJECTIVE: We sought to compare the frequency of BRAEs in patients receiving montelukast or placebo in a retrospective analysis of Merck clinical trial data. METHODS: An adverse experience database was constructed to include all double-blind, placebo-controlled trials of montelukast meeting prespecified criteria. BRAEs (described using the Medical Dictionary for Regulatory Activities controlled vocabulary dictionary) were prespecified to include any term in the Psychiatric Disorders System Organ Class, selected terms related to general disorders, and terms related to akathisia. Frequencies of BRAEs (overall, leading to study discontinuation, and/or serious) were summarized. Analyses estimated the odds ratios (ORs) for montelukast versus placebo based on the frequency of patients with BRAEs in each study. RESULTS: In total 35 adult and 11 pediatric placebo-controlled trials were included; 11,673 patients received montelukast, 8,827 received placebo, and 4,724 received active control. The frequency of patients with 1 or more BRAEs was 2.73% and 2.27% in the montelukast and placebo groups, respectively; the OR for montelukast versus placebo was 1.12 (95% CI, 0.93-1.36). The frequency of patients with a BRAE leading to study discontinuation was 0.07% and 0.11% in the montelukast and placebo groups, respectively (OR, 0.52; 95% CI, 0.17-1.51). The frequency of patients with a BRAE considered serious was 0.03% in both treatment groups. CONCLUSION: Reports of BRAEs were infrequent in clinical trials of montelukast. Those leading to study discontinuation or considered serious were rare. Frequencies were similar regardless of treatment group. PMID- 19815117 TI - Does exposure to cats and dogs decrease the risk of allergic sensitization and disease? PMID- 19815118 TI - Exaggerated IL-17 response to epicutaneous sensitization mediates airway inflammation in the absence of IL-4 and IL-13. AB - BACKGROUND: Atopic dermatitis (AD) is characterized by local and systemic T(H)2 responses to cutaneously introduced allergens and is a risk factor for asthma. Blockade of T(H)2 cytokines has been suggested as therapy for AD. OBJECTIVES: We sought to examine the effect of the absence of IL-4 and IL-13 on the T(H)17 response to epicutaneous sensitization in a murine model of allergic skin inflammation with features of AD. METHODS: Wild-type, IL4 knockout (KO), IL13 KO and IL4/13 double KO (DKO) mice were subjected to epicutaneous sensitization with ovalbumin (OVA) or saline and airway challenged with OVA. Systemic immune responses to OVA, skin and airway inflammation, and airway hyperresponsiveness were examined. RESULTS: OVA-sensitized DKO mice exhibited impaired T(H)2-driven responses with undetectable OVA-specific IgE levels and severely diminished eosinophil infiltration at sensitized skin sites but intact dermal infiltration with CD4(+) cells. DKO mice mounted exaggerated IL-17A but normal IFN-gamma and IL-5 systemic responses. Airway challenge of these mice with OVA caused marked upregulation of IL-17 mRNA expression in the lungs, increased neutrophilia in bronchoalveolar lavage fluid, airway inflammation characterized by mononuclear cell infiltration with no detectable eosinophils, and bronchial hyperresponsiveness to methacholine that were reversed by IL-17 blockade. IL-4, but not IL-13, was identified as the major T(H)2 cytokine that downregulates the IL-17 response in epicutaneously sensitized mice. CONCLUSION: Epicutaneous sensitization in the absence of IL-4/IL-13 induces an exaggerated T(H)17 response systemically and in lungs after antigen challenge that results in airway inflammation and airway hyperresponsiveness. PMID- 19815119 TI - Induction of long-lived allergen-specific plasma cells by mucosal allergen challenge. AB - BACKGROUND: Allergen-specific IgE antibodies are responsible for the pathogenesis of type I hypersensitivity. In patients with allergy, IgE titers can persist in the apparent absence of allergen for years. Seasonal allergen exposure triggers clinical symptoms and enhances allergen-specific IgE. Whether allergen-specific plasma cells originating from seasonal allergen exposures can survive and become long-lived is so far unclear. OBJECTIVE: We analyzed the localization and lifetimes of allergen-specific IgE-secreting, IgA-secreting, and IgG(1)-secreting plasma cells after allergen inhalation in an ovalbumin-induced murine model of allergic asthma. METHODS: Ovalbumin-specific IgG(1)-secreting, IgA-secreting, and IgE-secreting cells in lungs, spleen, and bone marrow were isolated and tested for antibody secretion by the ELISpot technique. Longevity of ovalbumin-specific plasma cells was determined by cyclophosphamide treatment, which depletes proliferating plasmablasts but leaves plasma cells untouched. Ovalbumin aerosol induced infiltrates in lungs were localized by confocal microscopy. RESULTS: Long lived ovalbumin-specific plasma cells were generated by systemic sensitization and survived in bone marrow and spleen, maintaining systemic ovalbumin-specific titers of IgG, IgA, and IgE. On inhalation of ovalbumin-containing aerosol, sensitized mice developed airway inflammation and more ovalbumin-specific IgG(1) secreting, IgA-secreting, and IgE-secreting cells in the lungs and in secondary lymphoid organs. These plasma cells joined the pool of ovalbumin-specific plasma cells in the bone marrow and became long-lived-that is, they are resistant to cyclophosphamide. Termination of ovalbumin inhalation depleted ovalbumin-specific plasma cells from the lungs, but they persisted in spleen and bone marrow. CONCLUSION: Our results show that inhalation of aerosolized allergen generates long-lived, allergen-specific IgG(1)-secreting, IgA-secreting, and IgE-secreting plasma cells that survive cytostatic treatment. PMID- 19815120 TI - Mumps resurgences in the United States: A historical perspective on unexpected elements. AB - In 2006 the United States experienced the largest nationwide mumps epidemic in 20 years, primarily affecting college dormitory residents. Unexpected elements of the outbreak included very abrupt time course (75% of cases occurred within 90 days), geographic focality (85% of cases occurred in eight rural Midwestern states), rapid upward and downward shift in peak age-specific attack rate (5-9 year olds to 18-24-year olds, then back), and two-dose vaccine failure (63% of case-patients had received two doses). To construct a historical context in which to understand the recent outbreak, we reviewed US mumps surveillance data, vaccination coverage estimates, and relevant peer-reviewed literature for the period 1917-2008. Many of the unexpected features of the 2006 mumps outbreak had been reported several times previously in the US, e.g., the 1986-1987 mumps resurgence had extremely abrupt onset, rural geographic focality, and an upward then-downward age shift. Evidence suggested recurrent mumps outbreak patterns were attributable to accumulation of susceptibles in dispersed situations where the risk of endemic disease exposure was low and were triggered when this susceptible population was brought together in crowded living conditions. The 2006 epidemic followed this pattern, with two unique variations: it was preceded by a period of very high vaccination rates and very low disease incidence and was characterized by two-dose failure rates among adults vaccinated in childhood. Data from the past 80 years suggest that preventing future mumps epidemics will depend on innovative measures to detect and eliminate build-up of susceptibles among highly vaccinated populations. PMID- 19815121 TI - Nebivolol: the somewhat-different beta-adrenergic receptor blocker. AB - Although its clinical use in Europe dates almost 10 years, nebivolol is a beta blocker that has been only recently introduced in the U.S. market. Like carvedilol, nebivolol belongs to the third generation of beta-blockers, which possess direct vasodilator properties in addition to their adrenergic blocking characteristics. Nebivolol has the highest beta(1)-receptor affinity among beta blockers and, most interestingly, it substantially improves endothelial dysfunction via its strong stimulatory effects on the activity of the endothelial nitric oxide synthase and via its antioxidative properties. Because impaired endothelial activity is attributed a major causal role in the pathophysiology of hypertension, coronary artery disease, and congestive heart failure, the endothelium-agonistic properties of nebivolol suggest that this drug might provide additional benefit beyond beta-receptor blockade. Although lesser beta blocker-related side effects have been reported in patients with chronic obstructive pulmonary disease or impotence taking nebivolol, side effects and contraindications overlap those of other beta-blockers. Clinically, this compound has been proven to have antihypertensive and anti-ischemic effects as well as beneficial effects on hemodynamics and prognosis in patients with chronic congestive heart failure. Further studies are now necessary to compare the benefit of nebivolol with that of other drugs in the same class and, most importantly, its prognostic impact in patients with hypertension. PMID- 19815122 TI - Improving the diagnosis of acute heart failure using a validated prediction model. AB - OBJECTIVES: We sought to derive and validate a prediction model by using N terminal pro-B-type natriuretic peptide (NT-proBNP) and clinical variables to improve the diagnosis of acute heart failure (AHF). BACKGROUND: The optimal way of using natriuretic peptides to enhance the diagnosis of AHF remains uncertain. METHODS: Physician estimates of probability of AHF in 500 patients treated in the emergency department from the multicenter IMPROVE CHF (Improved Management of Patients With Congestive Heart Failure) trial recruited between December 2004 and December 2005 were classified into low (0% to 20%), intermediate (21% to 79%), or high (80% to 100%) probability for AHF and then compared with the blinded adjudicated AHF diagnosis. Likelihood ratios were calculated and multiple logistic regression incorporated covariates into an AHF prediction model that was validated internally by the use of bootstrapping and externally by applying the model to another 573 patients from the separate PRIDE (N-Terminal Pro-BNP Investigation of Dyspnea in the Emergency Department) study of the use of NT proBNP in patients with dyspnea. RESULTS: Likelihood ratios for AHF with NT proBNP were 0.11 (95% confidence interval [CI]: 0.06 to 0.19) for cut-point values <300 pg/ml; increasing to 3.43 (95% CI: 2.34 to 5.03) for values 2,700 to 8,099 pg/ml, and 12.80 (95% CI: 5.21 to 31.45) for values > or =8,100 pg/ml. Variables used to predict AHF were age, pre-test probability, and log NT-proBNP. When applied to the external data by use of its adjudicated final diagnosis as the gold standard, the model appropriately reclassified 44% of patients by intermediate clinical probability to either low or high probability of AHF with negligible (<2%) inappropriate redirection. CONCLUSIONS: A diagnostic prediction model for AHF that incorporates both clinical assessment and NT-proBNP has been derived and validated and has excellent diagnostic accuracy, especially in cases with indeterminate likelihood for AHF. PMID- 19815123 TI - Diagnosing acute heart failure: the mathematician and the clinician. PMID- 19815124 TI - Altered myocardial substrate metabolism and decreased diastolic function in nonischemic human diabetic cardiomyopathy: studies with cardiac positron emission tomography and magnetic resonance imaging. AB - OBJECTIVES: This study was designed to evaluate myocardial substrate and high energy phosphate (HEP) metabolism in asymptomatic men with well-controlled, uncomplicated type 2 diabetes with verified absence of cardiac ischemia, and age matched control subjects, and to assess the association with myocardial function. BACKGROUND: Metabolic abnormalities, particularly an excessive exposure of the heart to circulating nonesterified fatty acids and myocardial insulin resistance are considered important contributors to diabetic cardiomyopathy in animal models of diabetes. The existence of myocardial metabolic derangements in uncomplicated human type 2 diabetes and their possible contribution to myocardial dysfunction still remain undetermined. METHODS: In 78 insulin-naive type 2 diabetes men (age 56.5 +/- 5.6 years, body mass index 28.7 +/- 3.5 kg/m(2), glycosylated hemoglobin A(1c) 7.1 +/- 1.0%; expressed as mean +/- SD) without cardiac ischemia and 24 normoglycemic control subjects (age 54.5 +/- 7.1 years, body mass index 27.0 +/- 2.5 kg/m(2), glycosylated hemoglobin A(1c) 5.3 +/- 0.2%), we assessed myocardial left ventricular (LV) function by magnetic resonance imaging, and myocardial perfusion and substrate metabolism by positron emission tomography using H(2)(15)O, carbon (11)C-palmitate, and 18-fluorodeoxyglucose 2-fluoro-2-deoxy-D glucose. Cardiac HEP metabolism was assessed by phosphorous P 31 magnetic resonance spectroscopy. RESULTS: In patients, compared with control subjects, LV diastolic function (E/A ratio: 1.04 +/- 0.25 vs. 1.26 +/- 0.36, p = 0.003) and myocardial glucose uptake (260 +/- 128 nmol/ml/min vs. 348 +/- 154 nmol/ml/min, p = 0.015) were decreased, whereas myocardial nonesterified fatty acid uptake (88 +/- 31 nmol/ml/min vs. 68 +/- 18 nmol/ml/min, p = 0.021) and oxidation (85 +/- 30 nmol/ml/min vs. 63 +/- 19 nmol/ml/min, p = 0.007) were increased. There were no differences in myocardial HEP metabolism or perfusion. No association was found between LV diastolic function and cardiac substrate or HEP metabolism. CONCLUSIONS: Patients versus control subjects showed impaired LV diastolic function and altered myocardial substrate metabolism, but unchanged HEP metabolism. We found no direct relation between cardiac diastolic function and parameters of myocardial metabolism. PMID- 19815125 TI - Potential clinical and economic consequences of noncardiac incidental findings on cardiac computed tomography. AB - OBJECTIVES: We sought to determine the incidence, clinical significance, and potential financial impact of noncardiac incidental findings (IF) identified with cardiac computed tomography (CT). BACKGROUND: Cardiac CT is gaining acceptance and may lead to the frequent discovery of extracardiac IF. METHODS: Consecutive patients undergoing cardiac CT had noncardiac structures evaluated after full field of view (32 to 50 cm) reconstruction. IF were categorized as clinically significant (CS), indeterminate, or clinically insignificant. Patient follow-up was performed by telephone, and verified with hospital records and/or communication with physicians. RESULTS: Of 966 patients (58 +/- 16 years of age, 55.4% men, >98% outpatients), 401 (41.5%) patients had noncardiac IF. A total of 12 (1.2%) patients had CS findings, and 68 (7.0%) patients had indeterminate findings. At follow-up (18.4 +/- 7.6 months), none of the indeterminate findings became CS. Although 3 patients with indeterminate findings were diagnosed with malignant lesions, they were unrelated to the IF. After adjusting for age, IF were not an independent predictor of noncardiac death. Noncardiac death and cancer death in patients with and without IF were not statistically different. One patient suffered a major complication related to the investigation of an IF. The total direct cost associated with investigating IF was Canadian $57,596 (U.S. $83,035). CONCLUSIONS: Although noncardiac IF are common, clinically significant or indeterminate IF are less prevalent. Rates of death were similar in patients with and without IF, and IF was not an independent predictor of noncardiac death. The investigation of IF is not without cost or risk. Larger studies are required to assess the potential mortality benefit of identifying IF. PMID- 19815126 TI - The dilemma of incidental findings on cardiac computed tomography. PMID- 19815127 TI - The year in non-ST-segment elevation acute coronary syndrome. PMID- 19815128 TI - Scimitar sign in a patient with an atrial septal defect: a comprehensive noninvasive assessment with transthoracic echocardiography and computed tomography. PMID- 19815129 TI - Safety and efficacy of pacemaker reuse in underdeveloped nations: a case series. PMID- 19815130 TI - The right ventricular outflow tract in arrhythmogenic right ventricular cardiomyopathy. PMID- 19815133 TI - Improve care with clinical practice guidelines. PMID- 19815134 TI - Board certification critical to preservation of a specialty. PMID- 19815136 TI - Cervical vertebrae maturation method: poor reproducibility. AB - INTRODUCTION: The cervical vertebrae maturation (CVM) method has been advocated as a predictor of peak mandibular growth. This method relies on the clinician's ability to determine the stage of maturation of the vertebrae. Careful examination of reports of this technique shows methodologic flaws that can lead to inflated levels of reproducibility. The purpose of this study was to evaluate the reproducibility of CVM stage determination by using a more stringent methodology. METHODS: Ten practicing orthodontists, trained in the CVM method, evaluated 30 individual and 30 pairs of cephalometric radiographs in 2 sessions to determine the CVM stage. Interobserver and intraobserver reliability was determined by using the Kendall coefficient of concordance and the weighted kappa statistic. RESULTS: All degrees of interobserver and intraobserver agreement were moderate (Kendall's W, 0.4-0.8). Interobserver agreement levels for CVM staging of the 10 orthodontists at both times were below 50%. Agreement improved marginally with the use of 2 longitudinal radiographs. Intraobserver agreement was only slightly better; on average, clinicians agreed with their own staging only 62% of the time. CONCLUSIONS: Based on these results, we cannot recommend the CVM method as a strict clinical guideline for the timing of orthodontic treatment. PMID- 19815138 TI - Geometric morphometric evaluation of cervical vertebrae shape and its relationship to skeletal maturation. AB - INTRODUCTION: Cervical vertebrae shape has been proposed as a diagnostic factor for assessing skeletal maturation in orthodontic patients. However, evaluation of vertebral shape is mainly based on qualitative criteria. Comprehensive quantitative measurements of shape and assessments of its predictive power have not been reported. Our aims were to measure vertebral shape by using the tools of geometric morphometrics and to evaluate the correlation and predictive power of vertebral shape on skeletal maturation. METHODS: Pretreatment lateral cephalograms and corresponding hand-wrist radiographs of 98 patients (40 boys, 58 girls; ages, 8.1-17.7 years) were used. Skeletal age was estimated from the hand wrist radiographs. The first 4 vertebrae were traced, and 187 landmarks (34 fixed and 153 sliding semilandmarks) were used. Sliding semilandmarks were adjusted to minimize bending energy against the average of the sample. Principal components analysis in shape and form spaces was used for evaluating shape patterns. Shape measures, alone and combined with centroid size and age, were assessed as predictors of skeletal maturation. RESULTS: Shape alone could not predict skeletal maturation better than chronologic age. The best prediction was achieved with the combination of form space principal components and age, giving 90% prediction intervals of approximately 200 maturation units in the girls and 300 units in the boys. Similar predictive power could be obtained by using centroid size and age. Vertebrae C2, C3, and C4 gave similar results when examined individually or combined. C1 showed lower correlations, signifying lower integration with hand-wrist maturation. CONCLUSIONS: Vertebral shape is strongly correlated to skeletal age but does not offer better predictive value than chronologic age. PMID- 19815140 TI - Use of cervical vertebral maturation to determine skeletal age. AB - INTRODUCTION: The purpose of this study was to evaluate the validity of the cervical vertebral maturation (CVM) method as an indicator of skeletal age in the circumpubertal period by correlating it to the hand-wrist method (HWM). METHODS: Hand-wrist and lateral cephalometric radiographs of 400 Chinese subjects were randomly selected. Their ages were 10 to 15 years for girls and 12 to 17 years for boys, so they were within the circumpubertal period. Skeletal ages were assessed according to the CVM method and the HWM. RESULTS: The CVM was significantly correlated with HWM skeletal age (Spearman r = 0.9521 [boys] and 0.9408 [girls]). All patients in cervical vertebral stage 3 of the CVM corresponded to stages MP3-FG or MP3-G (around the peak of the growth spurt) in the HWM. CONCLUSIONS: The CVM is a valid indicator of skeletal growth during the circumpubertal period, providing information for timing of growth modification. PMID- 19815142 TI - Percentiles relative to maxillary permanent canine inclination by age: a radiologic study. AB - INTRODUCTION: Few studies have investigated developmental norms for maxillary permanent canine eruption. In this observational cross-sectional study, we aimed to provide an age-related description of the percentiles relative to canine inclination in a large sample of nonorthodontic patients. Associations between inclination and sector were also analyzed. METHODS: Canine inclination and sector location were measured on 1020 panoramic radiographs obtained from subjects of white ancestry aged between 8 and 11 years not seeking orthodontic treatment. The total sample comprised 2037 canines. RESULTS: Canine inclination increases between 8 and 9 years and decreases between 9 and 11 years. The greatest value for each percentile is at 9 years. A linear model should be hypothesized for differences in canine inclination between 2 successive ages in correspondence to each percentile. The proportion of sector 2 canines decreases and that of sector 1 increases with age. In the same age group, the inclination generally decreases as the sector decreases. CONCLUSIONS: Percentiles by age show the average canine inclination in a certain population. Further studies are required to verify whether percentiles can be a diagnostic aid for determining normal canine inclination at a given age and for quantifying the risk of canine impaction or adjacent root resorption. PMID- 19815144 TI - Informed consent recall and comprehension in orthodontics: traditional vs improved readability and processability methods. AB - INTRODUCTION: Low general and health literacy in the United States means informed consent documents are not well understood by most adults. Methods to improve recall and comprehension of informed consent have not been tested in orthodontics. The purposes of this study were to evaluate (1) recall and comprehension among patients and parents by using the American Association of Orthodontists' (AAO) informed consent form and new forms incorporating improved readability and processability; (2) the association between reading ability, anxiety, and sociodemographic variables and recall and comprehension; and (3) how various domains (treatment, risk, and responsibility) of information are affected by the forms. METHODS: Three treatment groups (30 patient-parent pairs in each) received an orthodontic case presentation and either the AAO form, an improved readability form (MIC), or an improved readability and processability (pairing audio and visual cues) form (MIC + SS). Structured interviews were transcribed and coded to evaluate recall and comprehension. RESULTS: Significant relationships among patient-related variables and recall and comprehension explained little of the variance. The MIC + SS form significantly improved patient recall and parent recall and comprehension. Recall was better than comprehension, and parents performed better than patients. The MIC + SS form significantly improved patient treatment comprehension and risk recall and parent treatment recall and comprehension. Patients and parents both overestimated their understanding of the materials. CONCLUSIONS: Improving the readability of consent materials made little difference, but combining improved readability and processability benefited both patients' recall and parents' recall and comprehension compared with the AAO form. PMID- 19815146 TI - Long-term effects of orthodontic therapy on the maxillary dental arch and nasal cavity. AB - INTRODUCTION: Rapid maxillary expansion (RME) can enlarge the maxillary dental arch while increasing nasal cavity volume and nasal valve area, and decreasing upper airway resistance over time. However, the long-term effects of RME on arch morphology and nasal cavity geometry in patients treated with RME compared with the general population are unclear. METHODS: Three-dimensional morphometic analysis and acoustic rhinometry were used to evaluate the maxillary dental arches and nasal cavities in a post-RME-treatment group (n = 25) and a control group (n = 25). RESULTS: Palatal area and volume increased substantially after expansion and remained stable until posttreatment. Small increases in intermolar distance were not significant between the time points. Nasal airway resistance significantly decreased after RME and remained stable until posttreatment. Nasal cavity volume was stable during expansion and posttreatment. It increased significantly during expander stabilization and 9-12 months after expander removal. The minimal cross-sectional area significantly increased between the end of expansion and expander removal and remained stable until posttreatment. All measurements were comparable with those of the matched controls at posttreatment except for palatal area, which was smaller in the treatment group. CONCLUSIONS: RME is a powerful tool to normalize most of the variables investigated. Edgewise orthodontic treatment and a retention regimen that consisted of maxillary circumferential or traditional Hawley retainers met satisfactory standards to stabilize the achieved outcomes. Future studies should include morphometric, functional, and skeletal analyses so that the effects of growth and remodeling are better elucidated. PMID- 19815148 TI - Physical properties of root cementum: part 14. The amount of root resorption after force application for 12 weeks on maxillary and mandibular premolars: a microcomputed-tomography study. AB - INTRODUCTION: Orthodontic force magnitude is a primary factor in root resorption. Quantitative studies of root resorption after force application for 4 and 8 weeks have been conducted. In this study, we investigated the root surface topography and the amount of root resorption after the application of controlled light and heavy forces in a buccal direction for 12 weeks. In addition, the amounts of root resorption when controlled light and heavy forces were applied to the maxillary and mandibular first premolars were quantified. METHODS: Forty maxillary and mandibular first premolars were collected from 10 orthodontic patients (age range, 12.7-18.2 years; mean, 14.3 years). A light buccally directed orthodontic force of 25 g was applied to the experimental tooth on 1 side, and a heavy orthodontic force of 225 g was applied on the contralateral premolar. After 12 weeks of force application, the experimental teeth were extracted and scanned with the microcomputed tomography x-ray system. Resorption craters were analyzed with specially designed software for direct volumetric measurements. The tooth movements produced by light and heavy forces were also measured. RESULTS: There was individual variation in all comparisons. The light force produced significantly less root resorption than did the heavy force. The maxillary first premolars were more likely to suffer from orthodontically induced inflammatory root resorption than the mandibular first premolars (P = 0.036). There was a significant difference between buccal and lingual surfaces (P = 0.003), with greater root resorption on the buccal surface. The distribution pattern of the resorption cavities was greatest in the buccal-cervical, buccal-middle, lingual middle, and lingual-apical areas in both the light-force and heavy-force groups, corresponding with the pressure zones of tipping movement. The mean amount of tooth movement in the heavy-force group was almost twice as much as in the light force group. CONCLUSIONS: The volume of root resorption craters induced by buccally directed forces for 12 weeks on the maxillary and mandibular first premolars was directly proportional to the magnitude of the force. The maxillary premolars seemed to be more susceptible to orthodontic root resorption than did the mandibular premolars. PMID- 19815149 TI - Standard of care: why it is necessary. PMID- 19815150 TI - How can the specialty establish a standard of care? PMID- 19815151 TI - Effect of timing on the outcomes of 1-phase nonextraction therapy of Class II malocclusion. AB - INTRODUCTION: The aim of this cephalometric study was to evaluate the role of timing in relation to skeletal maturity on the outcomes of nonextraction comprehensive Class II therapy. METHODS: Three samples of patients with Class II Division 1 malocclusion were treated with headgear combined with fixed appliances and Class II elastics. Lateral cephalograms were taken of all subjects before therapy (T1) and at an average interval of 6 months after therapy (T2). The first sample (23 subjects) was treated before the pubertal growth spurt, the second sample (24 subjects) received therapy during the pubertal growth spurt, and the third sample (13 subjects) was treated at a postpubertal stage of development. The average T1 to T2 interval was approximately 30 months for all patients, with an average treatment duration of 24 months. Longitudinal observations of a group of 17 subjects with untreated Class II malocclusions were compared with the treated groups at the 3 skeletal maturation intervals with nonparametric statistics. RESULTS: Class II treatment before or during the pubertal growth spurt induced significant favorable skeletal changes (restricted maxillary advancement in prepubertal patients and enhanced mandibular growth in pubertal patients). Patients treated after the pubertal growth spurt had only significant dentoalveolar changes. CONCLUSIONS: The greatest amount of dentoskeletal correction of Class II malocclusion with 1-phase nonextraction treatment occurred in patients treated during the pubertal growth spurt. PMID- 19815152 TI - Effects of preoperative ibuprofen on pain after separator placement. AB - INTRODUCTION: Many patients experience pain with orthodontics, but there is no widely accepted standard of care for controlling orthodontic pain. Previous studies were inconclusive as to the most effective way to manage orthodontic pain. The purpose of this study was to assess the effectiveness in reducing pain of preemptive ibuprofen added to an ibuprofen regimen administered after separator placement. We also examined the contributions of psychological factors and sex to the experience of pain. METHODS: The subjects were randomly assigned to group A, 400 mg of ibuprofen 1 hour before separator placement (D1), 3 hours after placement (D2), and 7 hours after placement (D3); group B, placebo at D1, 400 mg of ibuprofen at D2 and D3; or group C, placebo at D1, D2, and D3. Before separator placement, the subjects completed 2 psychological surveys, a masticatory efficiency test, and a visual analog scale for expected pain and experienced pain during the masticatory efficiency test. After placement, the subjects recorded their actual pain, kept a pain diary for 24 hours after separator placement, and performed a 24-hour follow-up masticatory efficiency test. RESULTS: Group A, receiving ibuprofen before and after separator placement, experienced significantly less pain (P <0.05) at 6 hours, at bedtime, and at awakening on the second day. No contributions to pain were found for psychological factors or sex. CONCLUSIONS: Preemptive administration of analgesics should be recommended to orthodontic patients before separator placement. PMID- 19815153 TI - Three-dimensional orthodontic force measurements. AB - INTRODUCTION: Until recently, much of the orthodontic biomechanics literature was restricted to 2-dimensional experimental studies and, more recently, to assumption-based 3-dimensional computer modeling. There is little evidence in the literature regarding 3-dimensional experimental measurements and analysis of orthodontic force systems. METHODS: The purpose of this study was the design, construction, and validation of a laboratory-based human mouth model capable of accurately measuring forces and moments applied by orthodontic fixed appliances on all teeth in 1 arch. A high canine malocclusion was simulated, and forces and moments acting on the canine, lateral incisor, and premolar were measured with passive and conventional ligation. RESULTS: We were successful in building this human mouth model. The error in force measurements of the 14 transducers was 1.54%. The force system resulting from passive ligation brackets was considerably different from that of conventional ligation. CONCLUSIONS: This method will allow us, for the first time in the history of our specialty, to determine with great accuracy the forces acting on orthodontically treated teeth. Future research will focus on simulating many types of orthodontic clinical applications of full-fixed or partial-fixed appliances. PMID- 19815154 TI - Relationship between malocclusion, orthodontic treatment, and tooth wear. AB - INTRODUCTION: Although the association between orthodontic treatment and dental health has been studied previously, no studies have hitherto investigated whether the need for, or the receipt of, orthodontic treatment leads to differences in tooth wear in the long term. METHODS: In this retrospective, cross-sectional, association study, an index specifically designed for dental study casts was used to examine tooth wear in 307 adults aged 30 to 31 years. RESULTS: The men showed significantly higher levels of tooth wear than did the women. No significant differences in tooth wear in relation to need for or receipt of orthodontic treatment were recorded. Likewise, there appeared to be no significant association between tooth wear and reported intake of acidic drinks and foods. CONCLUSIONS: Neither the need for nor the provision of orthodontic treatment contributes to increased tooth wear. PMID- 19815155 TI - Further evidence for an association between mandibular height and the growth hormone receptor gene in a Japanese population. AB - INTRODUCTION: Craniofacial morphology has a significant genetic component. It has recently been reported that single nucleotide polymorphisms (SNPs) in the growth hormone receptor gene (GHR) are associated with mandibular height. To confirm these findings, we genotyped SNPs in healthy Japanese subjects, about 1.7 times the number analyzed in previous reports. METHODS: By using 5 SNPs in exon 10 of the GHR, we studied the relationships between genotypes and craniofacial linear measurements. The allelic frequencies in a multi-ethnic population (Han Chinese, African American, European American, and Hispanic) were also observed. RESULTS: There was a significant association between SNPs and mandibular ramus height (P = 0.029), confirming our previous report of an association between polymorphism P561T and mandibular ramus height. Moreover, the SNP, C422F, was in linkage disequilibrium with P561T. Subjects with genotype CC of polymorphism P561T and genotype GG of polymorphism C422F had significantly greater mandibular ramus height than those with genotypes CA and GT. Four of the 5 SNPs were found almost exclusively in Asians, with the frequencies in other populations extremely small. CONCLUSIONS: Our results indicate that the GHR polymorphisms P561T and C422F are associated with mandibular ramus height in Japanese population and suggest that the SNPs of the GHR associated with the Japanese are likely to be different in other ethnic groups. This might partly explain the differing craniofacial morphology among different ethnicities. PMID- 19815156 TI - Association of functional gene polymorphism IL-1beta in patients with external apical root resorption. AB - INTRODUCTION: External apical root resorption (EARR) is a frequent complication of orthodontic treatment. Interleukin 1beta (IL-1beta) is a potent stimulus for bone resorption and osteoclastic cell recruitment during orthodontic tooth movement. The purpose of this study was to assess the association between a polymorphism in the IL-1beta genes and EARR during orthodontic treatment. METHODS: The sample included 61 Brazilian orthodontic patients, divided into 2 groups according to the presence (affected group, n = 23) or absence (control group, n = 38) of EARR in the central and lateral maxillary incisors in the posttreatment period. DNA was obtained from buccal swab cells. The polymorphism was analyzed by the polymerase chain reaction followed by digestion with restriction enzyme. The polymerase chain reaction products were analyzed in 10% polyacrylamide gel and stained with silver. RESULTS: There were significant statistical differences (P <0.05) among the frequencies of the alleles and genotypes of the IL-1beta gene polymorphism between the affected and unaffected groups, suggesting that allele 1 predisposed the subjects to EARR (odds ratio = 4.0). CONCLUSIONS: The polymorphism of the IL-1beta gene is associated with root resorption in the studied population. PMID- 19815157 TI - Fluoride release and cariostatic potential of orthodontic adhesives with and without daily fluoride rinsing. AB - INTRODUCTION: In this study, we aimed to evaluate the fluoride-release profiles and caries lesion development in an enamel model with brackets cemented with 4 orthodontic adhesives with and without daily fluoride exposure. METHODS: Four orthodontic adhesives (Ketac Cem mu, 3M ESPE, Seefeld, Germany; Fuji Ortho LC, GC Corporation, Tokyo, Japan; Light-Bond, Reliance Orthodontic Products, Itasca, Ill; and Transbond XT, 3M Unitek, Monrovia, Calif) were used. Brackets were bonded on bovine enamel with each adhesive (n = 10) and subjected to alternate cycles of demineralizing (pH 4.55) and remineralizing (pH 6.8) solutions. Unbracketed enamel samples served as a reference. Five samples from each group were immersed in a fluoride mouth rinse (250 ppm fluoride) for 1 minute each day (test groups). Fluoride release was measured at regular intervals over 28 days. The mineral distribution of peribracket enamel after 28 days was quantified by transversal microradiographs. RESULTS: Fluoride-release profiles of Ketac Cem mu, Fuji Ortho LC, and Light-Bond were high for the first 24 hours and reached a constant level after 2 weeks. Fuji Ortho LC released significantly more fluoride than did the other adhesives in both the control and test groups (P <0.01, repeated measures ANOVA and Bonferroni test). Enamel bonded with Fuji Ortho LC had significantly shallower lesions and less mineral loss (P <0.01, 2-way ANOVA, and Tukey HSD). CONCLUSIONS: Bonding of orthodontic brackets with Fuji Ortho LC resulted in less peribracket enamel demineralization with and without daily fluoride rinsing, mainly due to its better fluoride-release profile. In contrast, Transbond XT and Light-Bond offered few cariostatic effects to the enamel. PMID- 19815158 TI - Craniofacial characteristics of patients with heart disease. AB - INTRODUCTION: Children with congenital heart disease commonly experience delayed growth. Studies comparing growth and development of these children generally found that they have decreased height and weight, but with no information on craniofacial growth. The aim of this study was to evaluate the craniofacial characteristics of patients with heart disease. METHODS: We used lateral head cephalograms of 131 patients, aged 8 to 12, attending the Department of Pediatrics of the Hospital of the State University of Rio de Janeiro in Brazil on a routine basis. The study group comprised 50 boys and 38 girls with congenital heart disease. The control group included 20 boys and 23 girls with no heart disease. The skeletal, dental, growth, esthetic, and vertical patterns were analyzed with several cephalometric measurements. The chi-square test was applied to verify a possible association between sex and heart disease, and the Student t test was used to verify differences between the groups of patients with and without heart disease. RESULTS: Heart disease is not sex-related (chi-square = 0.267). But some measurements had sex differences in the groups with and without heart diseases. The Student t test showed significant differences between the groups in the SNA angle measurement in male patients (P = 0.032), and in ANB angle (P = 0.002) and the relationship of the lower lip to the Steiner line (P = 0.034) in the girls. CONCLUSIONS: Skeletal patterns were characterized by maxillary protrusion in the boys with heart disease and by increased ANB angles in the girls without heart disease. The esthetic pattern showed a more pronounced lower lip in the girls without heart disease. However, from a clinical point of view, these were slight and irrelevant alterations. The results suggest that patients suffering from heart disease have similar craniofacial characteristics to those without this condition. PMID- 19815159 TI - Asymmetry of the lips of orthognathic surgery patients. AB - INTRODUCTION: The purpose of this study was to examine asymmetry of the lips in patients with jaw deformity and facial asymmetry. METHODS: Pretreatment and posttreatment frontal facial photographs of 17 patients who had mandibular prognathism with facial asymmetry were analyzed. Linear and angular measurements were made on the photographs to assess the asymmetry of the lips. The upper and lower vermilion borders of the lips were divided into quadrants according to the y-axis, and each was measured. In addition, skeletal deviation (position of menton) was determined from the frontal cephalograms. RESULTS: The pretreatment linear and angular measurements showed asymmetry of the lips, but this significantly improved after treatment, and the lip form was almost symmetrical. The areas of vermilion border in the upper and lower lips became almost even during treatment. The ratio of the areas on the 2 sides was nearly 1.00. There was little correlation between the horizontal skeletal changes of menton and lip form. CONCLUSIONS: Asymmetry of the lips caused by deviation of the mandible can be almost completely corrected by orthognathic treatment. PMID- 19815160 TI - Change of lip-line cant after 1-jaw orthognathic surgery in patients with mandibular asymmetry. AB - INTRODUCTION: The purpose of this study was to investigate the change of lip-line cant (LLC) after 1-jaw orthognathic surgery in mandibular asymmetry patients. METHODS: Preoperative and postoperative data of 22 patients having 1-jaw orthognathic surgery, with menton deviation over 2 degrees before the surgery, were our subjects. LLC was measured in the preoperative and postoperative frontal photographs, and its change was correlated with various craniofacial measurements obtained from preoperative and postoperative frontal cephalograms and maxillofacial 3-dimensional computed tomography images. RESULTS: Although these subjects had 2.4 degrees of LLC on average before surgery, LLC improved to 0.5 degrees after surgery, and the change (1.9 degrees ) was statistically significant. In the correlation analysis, preoperative LLC showed positive correlations with menton deviation and mandibular anterior occlusal plane cant. In the correlation analysis of LLC change, it had positive correlations with preoperative LLC and mandibular anterior occlusal plane cant and preoperative and postoperative change of menton deviation. CONCLUSIONS: These results suggest that LLC is present with chin deviation, even without significant maxillary canting, and can be improved considerably by 1-jaw surgery alone. PMID- 19815161 TI - Comparison of corticotomy-facilitated vs standard tooth-movement techniques in dogs with miniscrews as anchor units. AB - INTRODUCTION: One method used to accelerate orthodontic tooth movement is the corticotomy-facilitated (CF) technique. The purposes of this study were to (1) identify the effect of the CF technique on orthodontic tooth movement compared with the standard technique, and (2) explore the histologic basis of the difference between the 2 techniques. METHODS: Six dogs, aged 6 to 9 months, were used in this study. Extraction of the maxillary second premolar and miniscrew placement were done bilaterally in the maxilla. On the right side, the corticotomy was performed. The first premolars were distalized against the miniscrews with nickel-titanium coil springs on both sides. One dog was killed each week after orthodontic force application. RESULTS: The first premolar on the CF side moved significantly more rapidly (P <0.05). Histologic findings showed more active and extensive bone remodeling on both the compressive and tension sides in the CF group. CONCLUSIONS: The CF technique doubled the rate of orthodontic tooth movement. Histologically, the more active and extensive bone remodeling in the CF group suggested that the acceleration of tooth movement associated with corticotomy is due to increased bone turnover and based on a regional acceleratory phenomenon. PMID- 19815162 TI - Efficiency of a skeletonized distal jet appliance supported by miniscrew anchorage for noncompliance maxillary molar distalization. AB - INTRODUCTION: Conventional anchorage appliances rely exclusively on intraoral anchorage for noncompliance molar distalization. The partial coverage of the palate, in particular, often results in compromised oral hygiene. An innovative alternative combines a skeletonized distal jet appliance with 2 paramedian miniscrews for additional anchorage. The objectives of this study were to investigate the suitability of the skeletonized distal jet for translatory molar distalization and to check the quality of the supporting anchorage setup. METHODS: Two paramedian miniscrews (length, 8-9 mm; diameter, 1.6 mm) were placed into the anterior area of the palate in 10 patients. Skeletonized distal jet appliances fitted with composite to the first premolars and the collars of the miniscrews were used for bilateral molar distalization, and the coil springs were activated with a distalization force of 200 cN on each side. RESULTS: The study confirmed the suitability of the appliance for translatory molar distalization (3.92 +/- 0.53 mm) with slight mesial inward rotation (on average, 8.35 degrees +/- 7.66 degrees and 7.88 degrees +/- 5.50 degrees ). The forces acting reciprocally on the anchorage setup were largely absorbed by the anchorage unit involving 2 anchorage teeth and 2 miniscrews. Significant anchorage loss, in the form of first premolar mesialization of 0.72 +/- 0.78 mm, was found. CONCLUSIONS: The skeletonized distal jet appliance supported by additional miniscrew anchorage allows translatory molar distalization. Although the anchorage design combining 2 miniscrews at a paramedian location and the periodontium of 2 anchorage teeth does not offer the quality of stationary anchorage, it achieves greater molar distalization in total sagittal movement than conventional anchorage designs with an acrylic button. PMID- 19815163 TI - Titanium miniplate as anchorage to close a premolar space by means of mesial movement of the maxillary molars. AB - We report the orthodontic treatment of a woman who was congenitally missing teeth 15, 47, and 48. A titanium miniplate was used as anchorage to close the space of the missing premolar by mesial movement of the maxillary posterior teeth into the space of tooth 15. Teeth 16 and 17 were moved 7.4mm mesially, until the proximal contact of tooth 16 with tooth 14 was detected clinically. Good esthetic and dental results were achieved after 34 months of active treatment. Miniplates can be used as orthodontic anchorage to move maxillary molars forward. PMID- 19815164 TI - Treatment of a horizontal open bite with an invisible multiloop appliance in a girl with tooth trauma. AB - This case report presents the treatment of a girl with a Class II horizontal open bite and severe crowding with an invisible appliance. She had been in a severe traffic accident at 5 years 10 months of age. Her teeth, especially the mandibular right lateral incisor, canine, and first premolar had ankylosed, and their roots were severely resorbed and fused to the alveolar bone. Orthodontic treatment started at the age of 12 years. The mandibular left first premolar and both maxillary first premolars were extracted to reduce overjet and crowding. The left first premolar was extracted and transplanted to the extracted position of the right first premolar. Because the patient wanted an invisible appliance, we selected the Fujita lingual bracket system for her treatment. During the final stage of treatment, a multiloop mushroom archwire was placed to correct the open bite in the maxillary arch with vertical elastics. Six years after retention, her occlusion remained stable, and the transplanted premolar was functioning normally. PMID- 19815165 TI - Use of technology in the orthodontic practice: a day in the life. PMID- 19815166 TI - An interesting and provocative issue to close out the 10th volume of CRM. PMID- 19815167 TI - Early and mid-term clinical outcomes with the CATANIA coronary stent system vs. bare metal stents in patients with coronary artery disease. AB - BACKGROUND: The potential for the CATANIA (CAT) stent to be an alternative to both bare-metal stents (BMS) and drug-eluting stents (DES) has been recently demonstrated in the Assessment of The LAtest Non-Thrombogenic Angioplasty stent (ATLANTA) first-in-human study. The aim of the present study was to compare short term outcomes of patients treated with the CAT stent with those treated with BMS. METHODS: Based on an internal registry, the 30-day and 6-month risk-adjusted outcomes for patients who received the CAT stent (n=254) were compared against outcomes of a historical cohort of patients who received BMS (n=552) between January 2001 and December 2001. RESULTS: At 30 days, use of BMS vs. the CAT stent resulted in borderline significant differences with respect to major adverse cardiac and cerebrovascular events (MACCE) and cardiac death or myocardial infarction. At 6 months, BMS showed a statistically significant higher adjusted risk of MACCE (HR 2.79, 95% CIs 1.20-6.48, P=.017) and no differences with respect to the subcomponent end points. The cumulative incidence of definite stent thrombosis (Academic Research Consortium defined) at 6 months was 0.39% for the CAT stent and 2.35% for the BMS. CONCLUSIONS: This study confirms the favorable early and mid-term safety profile and the high-level efficacy of the CAT stent in the treatment of de novo coronary lesions seen in the ATLANTA trial. The use of stents with a nanothin Polyzene-F surface treatment provided improved results with respect to BMS and lower risk of acute and subacute stent thrombosis. PMID- 19815168 TI - Safety and efficacy of carotid artery stenting in persons aged 80 years and above. AB - Carotid artery stenting (CAS) remains a controversial procedure in spite of an emerging abundance of trial data showing both safety and efficacy. CAS is under significant scrutiny by the Centers for Medicare and Medicaid Services as it relates to persons aged 80 years and above. We report 1-year follow up both clinically and with ultrasonography in a small cohort of patients with an average age of 85 years. PMID- 19815169 TI - Prophylactic use of manual thrombectomy in ST-segment elevation myocardial infarction. AB - OBJECTIVE: We sought to evaluate the effects of manual thrombectomy on myocardial reperfusion performed during percutaneous coronary intervention (PCI) for ST segment elevation myocardial infarction (STEMI). BACKGROUND: Complete reperfusion after primary PCI is compromised by the presence of intraluminal thrombus. Thus effective and safe extraction of thrombus in a timely fashion is important for successful reperfusion. METHODS: Thirty-two patients (age 51+/-12 years, males 78%) with STEMI and angiographic evidence of intraluminal thrombus underwent thrombectomy during an 18-month period. Thrombectomy was performed after the presence of thrombus was confirmed angiographically by the operator either before or after primary angioplasty. Thrombectomy was performed using the 6F Export Aspiration Catheter (Medtronic Corporation, Santa Rosa, CA, USA). Myocardial reperfusion using Thrombolysis in Myocardial Infarction (TIMI) flow and myocardial blush grade was assessed by two independent observers. RESULTS: The infarct-related artery was left anterior descending (59%), right coronary artery (19%), saphenous venous graft (19%), or left circumflex artery (3%). The coronary lesion was Type B in 62% and Type C in 37% patients, with an average length of 18.2+4.6 mm and reference vessel diameter of 3.2+/-0.4 mm. The preprocedural TIMI flow was 0 in 62%, 1 in 12%, 2 in 22%, and 3 in 3% of patients. The postprocedural TIMI flow was 0 in 3%, 1 in 6%, 2 in 25%, and 3 in 56% of patients. The postprocedural myocardial blush grade was 0 in 6%, 1 in 9%, 2 in 35%, and 3 in 48% of patients. The in-hospital mortality was 0 and the 30-day mortality was 3%. CONCLUSION: Manual thrombectomy using an Export catheter is safe and effective in establishing myocardial reperfusion after STEMI. PMID- 19815170 TI - Preliminary observations using optical coherence tomography to assess neointimal coverage of a metal stent in a porcine model. AB - BACKGROUND: Concerns surrounding late stent thrombosis have prompted the development of novel imaging techniques to assess neointimal coverage. Recent clinical studies have evaluated optical coherence tomography (OCT) to evaluate neointimal coverage, but pathologic correlation in an animal model is lacking. We assessed the hypothesis that OCT could accurately assess early neointimal coverage in a porcine model. METHODS: OCT imaging of bare metal stents in each coronary artery was performed at implantation (n=6), Day 4 (n=3), and Day 20 (n=3), and images were evaluated at three cross-sections per stented segment. Neointimal strut coverage was categorized by OCT as covered or uncovered, and neointimal thickness was determined (Day 20). Pathological correlation was obtained using scanning electron microscopy (SEM) to assess strut coverage (Day 4) and histomorphometry to quantify neointimal thickness (Day 20). RESULTS: At Day 4, OCT imaging detected 28 (26%) of 109 uncovered struts, and the ratio of uncovered/total strut area by SEM was 31%. All imaging modalities showed complete coverage at Day 20. Mean (+/-SE) neointimal thickness at Day 20 was 109+/-6 microm by OCT (n=116 struts) and 93+/-5 microm by pathology (n=68). Mean neointimal thickness on a segment-by-segment basis determined by OCT correlated with mean histomorphometric analysis (Reviewer 1: r=.74, P=.092 and Reviewer 2: r=0.60, P=.212). CONCLUSIONS: Day 4 represents an important time point for the assessment of early neointimal coverage in the porcine model. OCT imaging accurately assesses the extent and thickness of early neointimal coverage with good pathologic correlation. OCT represents a promising imaging modality for the in vivo assessment of neointimal coverage. PMID- 19815171 TI - Late-acquired incomplete stent apposition: morphologic characterization. AB - Incomplete stent apposition (ISA) is a lack of contact between stents and the underlying vessel wall, best described by intravascular ultrasound (IVUS). Late acquired incomplete apposition, defined as complete stent apposition at the time of procedure but ISA at follow-up, is an unusual IVUS finding reported in intracoronary brachytherapy, bare-metal stent (BMS), and drug-eluting stent (DES) implantation. Late-acquired ISA is observed relatively more frequently with DES implantation compared with BMS implantation. Possible mechanisms of this phenomenon include focal/extensive vascular remodeling and dissolution of thrombus. While there are conflicting reports regarding the possible impact of this IVUS finding on clinical outcomes, recent reports of DES have suggested its possible association with late adverse cardiac events including late stent thrombosis. In this paper, we review the incidence, location, underlying pathology, and possible clinical sequelae of late-acquired ISA, primarily focusing on that of DES. PMID- 19815172 TI - Biomaterial optimization in a percutaneous aortic valve stent using finite element analysis. AB - BACKGROUND: Vascular support structures are important devices for treating valve stenosis. Large population of patients are treated for valvular disease and the principal mode of treatment is the use of percutaneous valvuloplasty. Stent devices are proving to be an improved technology in minimal invasive cardiac surgery. This technology now accounts for 20% of treatments in Europe. This new technology provides highly effective results at minimal cost and short duration of hospitalization. METHODS: This paper discusses the design and finite element analysis (FEA) of a percutaneous aortic valve stent. The stent design was modeled and subjected to FEA. The improved model design was carried out to meet the functional and surgical requirements. Analysis was done with different materials with loads ranging from 50 to 73 kgf/mm(2). These forces were selected because these values are far greater than the normal human blood pressure which ranges from 10 to 16 kPa. It was also to understand the mechanical behavior of different stent materials under such high pressures. RESULTS: A stent model was generated and its physical, mechanical, and behavioral properties were studied. Finite element analysis and simulation of the model help the designer to optimize the geometry suitable for performance during and after implantation. The design objective for the stent is to have long-term durability, low thrombogenicity, and resistance to migration and paravalvular leak. CONCLUSION: The analysis performed in this paper may aid in understanding the stent's tolerable pressure ranges in comparison with the physiological pressures exerted by the heart and cardiac blood flow during abnormal cardiovascular conditions. It may also help in finding the material that will best suit the stent. PMID- 19815173 TI - Multiple coronary artery aneurysm formation one year and four years after sirolimus-eluting coronary stent implantation. AB - A 61 years old male patient having the complaint of angina pectoris underwent coronary angiography that revealed critical luminal stenosis at left anterior descending artery (LAD) and diagonal branch. Percutaneous transluminal coronary angioplasty was performed and sirolimus eluting stents (CYPHER; Johnson & Johnson -Cordis) 2.75 x 10 mm and 2.75 x 10 mm were deployed respectively. One year after the procedure coronary angiography revealed coronary artery aneurysm (CAA) formation in diagonal branch and mild ectasia in LAD stent area. Last coronary angiography four years and three months after initial intervention showed multiple CAA at diagonal branch and LAD. This case report is an example of late formation of CAA by sirolimus-coated coronary stent. PMID- 19815175 TI - Fibromuscular dysplasia of ostial left main coronary artery depicted by intravascular ultrasound. PMID- 19815174 TI - Funneling: enhancing results of small-vessel stenting. AB - Until October 2008, coronary drug-eluting stents with a diameter of < or =2.5 mm were not approved by the FDA. Target vessels of < or =2 mm in diameter pose a major challenge in view of high restenosis rates when stented bare metal stents (BMS) are used and distal edge dissection when oversized stents are deployed. Described is a method ("funneling") to optimize stenting of small vessels. This strategy combines stenting the distal part of the lesion with short (8-12 mm length) 2-mm bare metal stent while stenting with a larger-diameter (> or =2.5 mm) drug-eluting stent (DES) with considerable DES-BMS overlap (leaving only the distal 4-6 mm of the BMS not overlapped by a DES). The two stents create a funnel that is for the most part drug eluting. With funneling, both drug elution and larger diameter are attained, minimizing both edge dissection and restenosis. PMID- 19815176 TI - Macromolecular trafficking and immune evasion in african trypanosomes. AB - Intracellular trafficking is a major mechanism contributing to maintenance of the surface composition in most eukaryotic cells. In the case of unicellular eukaryotic pathogens, the surface also represents the host-parasite interface. Therefore, the parasite surface is both a critical player in immune recognition, from the host's point of view, or in immune evasion, from the pathogen's point. The African trypanosomes are remarkable in dwelling throughout their period in the mammalian host within the bloodstream and tissue spaces, and have evolved several mechanisms that facilitate chronic infection. Here, we discuss current understanding of intracellular trafficking pathways of trypanosomes, and relate these processes to immune evasion strategies by the parasite and avoidance of immune responses from the host. PMID- 19815178 TI - Genotype-phenotype mapping developmental biology confronts the toolkit paradox. AB - The quest to understand the relationship between an organism's DNA sequence and three-dimensional form is an interdisciplinary task, integrating diverse fields of the life sciences. The relevance of the metaphor of a genotype-phenotype map is explored from a developmental perspective, in light of the recent concept of a "molecular toolkit" of protein-coding genes, and the widespread view that analyzing the logic and mechanics of gene regulation at multiple levels is key to explaining how morphology is genetically encoded. We discuss the challenges of decoding genomes despite variable genetic backgrounds, the dynamically changing physical and molecular contexts of the internal environment during development, and the impact of external forces on morphogenesis. PMID- 19815179 TI - Role of spindle asymmetry in cellular dynamics. AB - The mitotic spindle is mostly perceived as a symmetric structure. However, in many cell divisions, the two poles of the spindle organize asters with different dynamics, associate with different biomolecules or subcellular domains, and perform different functions. In this chapter, we describe some of the most prominent examples of spindle asymmetry. These are encountered during cell-cycle progression in budding and fission yeast and during asymmetric cell divisions of stem cells and embryos. We analyze the molecular mechanisms that lead to generation of spindle asymmetry and discuss the importance of spindle-pole differentiation for the correct outcome of cell division. PMID- 19815177 TI - Biological and biophysical properties of vascular connexin channels. AB - Intercellular channels formed by connexin proteins play a pivotal role in the direct movement of ions and larger cytoplasmic solutes between vascular endothelial cells, between vascular smooth muscle cells, and between endothelial and smooth muscle cells. Multiple genetic and epigenetic factors modulate connexin expression levels and/or channel function, including cell-type independent and cell-type-specific transcription factors, posttranslational modifications, and localized membrane targeting. Additionally, differences in protein-protein interactions, including those between connexins, significantly contribute to both vascular homeostasis and disease progression. The biophysical properties of the connexin channels identified in the vasculature, those formed by Cx37, Cx40, Cx43 and/or Cx45 proteins, are discussed in this chapter in the physiological and pathophysiological context of vessel function. PMID- 19815180 TI - Cell adhesion in amphibian gastrulation. AB - The amphibian gastrula can be regarded as a single coherent tissue which folds and distorts itself in a reproducible pattern to establish the embryonic germ layers. It is held together by cadherins which provide the flexible adhesion required for the massive cell rearrangements that accompany gastrulation. Cadherin expression and adhesiveness increase as one goes from the vegetal cell mass through the anterior mesendoderm to the chordamesoderm, and then decrease again slightly in the ectoderm. Together with a basic random component of cell motility, this flexible, differentially expressed adhesiveness generates surface and interfacial tension effects which, in principle, can exert strong forces. However, conclusive evidence for an in vivo role of differential adhesion-related effects in gastrula morphogenesis is still lacking. The most important morphogenetic process in the amphibian gastrula seems to be intercellular migration, where cells crawl actively across each other's surface. The crucial aspect of this process is that cell motility is globally oriented, leading for example to mediolateral intercalation of bipolar cells during convergent extension of the chordamesoderm or to the directional migration of unipolar cells during translocation of the anterior mesendoderm on the ectodermal blastocoel roof. During these movements, the boundary between ectoderm and mesoderm is maintained by a tissue separation process. PMID- 19815181 TI - Molecular and cell biology of testicular germ cell tumors. AB - Although testicular germ cell tumors (TGCTs) are relatively uncommon, they are particularly important as they tend to affect children and young men, representing the most common tumor in male aged from 20 to 40years. TGCTs are a heterogeneous group of tumors, with specific peculiarities reflecting on epidemiologic distribution and clinic-pathological features. TGCTs show a high cure rates in both seminomas and nonseminomas and represent the model of a curable neoplasia: sensitive serum tumor markers, accurate prognostic classification, contribute to a high effectiveness of cancer therapy. However, up to 30% of patients diagnosed with metastatic nonseminomas do not achieve a durable remission, and in metastatic teratomas cisplatin-based treatment resistance has been observed. These different prognostic and therapeutic features of TGCTs highlight the need for a better understanding of the molecular biology of TGCT, that could help to improve disease management and to tailor aggressiveness of treatment to the severity of the prognosis. PMID- 19815183 TI - The Operational Demand Evaluation Checklist (ODEC) of workload for railway signalling. AB - This paper is concerned with the interpretation and assessment of mental workload, and in particular assessment of the load imposed by the work system. It highlights a framework created to direct the development of workload assessment tools capable of assessing the dimensions most relevant to the population being studied, in our case railway signallers. A tool to capture the operational demands on the rail signaller was required to evaluate the load from the system they operated. This paper justifies the need for, and describes the development of, the Operational Demand Evaluation Checklist (ODEC), using techniques like repertory grid with active signallers. The practical experience of the development, evaluation, live use and validation of ODEC is discussed and the paper concludes by suggesting that the approach could be adopted to interpret the concept of workload in other work domains. PMID- 19815184 TI - Decoupling implicit measures of pleasant and unpleasant social attitudes. AB - Social attitudes are integral to understanding a wide range of pathological states. The present study adapted the Implicit Association Test, a widely used implicit measure of attitudes, for understanding social attitudes and behavior. In a first study, data from a traditional "Bipolar" IAT and our modified "Unipolar" pleasant and unpleasant IATs were compared in their associations with explicit measures of social attitudes and behavior. In a second study, we examined the relationship between implicit measures of social attitudes and social behavior during a laboratory procedure. Collectively, the present data support the uncoupling of pleasant and unpleasant valences when using implicit measures to understand social attitudes. PMID- 19815182 TI - Polarity proteins and cell-cell interactions in the testis. AB - In mammalian testes, extensive junction restructuring takes place in the seminiferous epithelium at the Sertoli-Sertoli and Sertoli-germ cell interface to facilitate the different cellular events of spermatogenesis, such as mitosis, meiosis, spermiogenesis, and spermiation. Recent studies in the field have shown that Rho GTPases and polarity proteins play significant roles in the events of cell-cell interactions. Furthermore, Rho GTPases, such as Cdc42, are working in concert with polarity proteins in regulating cell polarization and cell adhesion at both the blood-testis barrier (BTB) and apical ectoplasmic specialization (apical ES) in the testis of adult rats. In this chapter, we briefly summarize recent findings on the latest status of research and development regarding Cdc42 and polarity proteins and how they affect cell-cell interactions in the testis and other epithelia. More importantly, we provide a new model in which how Cdc42 and components of the polarity protein complexes work in concert with laminin fragments, cytokines, and testosterone to regulate the events of cell-cell interactions in the seminiferous epithelium via a local autocrine-based regulatory loop known as the apical ES-BTB-basement membrane axis. This new functional axis coordinates various cellular events during different stages of the seminiferous epithelium cycle of spermatogenesis. PMID- 19815185 TI - Addition of amines and carbon nucleophiles to vinyl sulfone-modified 6-deoxy-hex 3-enopyranoside: a case of nucleophile dependent diastereoselectivity. AB - Reactions of amines and carbon nucleophiles with 4-sulfonyl-hex-3-enopyranoside generate a range of C-3 amino- and C-3 branched-chain sugars, which are analogues of 3-amino-3,6-dideoxy sugars and 3-C-branched-chain-3,6-dideoxy sugars. The diastereoselectivity of addition reaction is nucleophile dependent; while both nitrogen and carbon nucleophiles added in cis-fashion, amines generated C3-C4 trans-diaxial products (gulo-derivatives), and carbon nucleophiles afforded C3-C4 trans-diequatorial products (gluco-analogues). PMID- 19815186 TI - Structural analysis of an immunoenhancing heteropolysaccharide isolated from the green (unripe) fruits of Solenum melongena (Brinjal). AB - A water-soluble polysaccharide (SMPS) isolated from the aqueous extract of the green (unripe) fruits of Solenum melongena (Brinjal) contains D-galactose, D methyl galacturonate, 3-O-acetyl D-methyl galacturonate, and L-arabinose in a molar proportion of nearly 1:1:1:1. Structural investigation of the polysaccharide was carried out using total acid hydrolysis, methylation analysis, periodate oxidation study, and NMR studies ((1)H, (13)C, DQF-COSY, TOCSY, NOESY, ROESY, HMQC, and HMBC). On the basis of the above experiments the structure of the repeating unit of the polysaccharide (SMPS) was established as -->5)-alpha-L Araf-(1-->4)-beta-D-Galp-(1-->2)-alpha-D-3-O-Ac-GalpA6Me-(1-->2)alpha-D-galpA6Me (1--> This molecule showed splenocyte and thymocyte activations. PMID- 19815187 TI - Exemplar similarity and rule application. AB - We report four experiments examining effects of instance similarity on the application of simple explicit rules. We found effects of similarity to illustrative exemplars in error patterns and reaction times. These effects arose even though participants were given perfectly predictive rules, the similarity manipulation depended entirely on rule-irrelevant features, and attention to exemplar similarity was detrimental to task performance. Comparison of results across studies suggests that the effects are mandatory, non-strategic and not subject to conscious control, and as a result, should be pervasive throughout categorization. PMID- 19815188 TI - Ageing affects brain activity in highly educated older adults: An ERP study using a word-stem priming task. AB - In this event-related evoked potentials (ERP) study, the neural correlates of a group of highly educated older adults were compared with those of a group of young adults while performing a word-stem completion priming task under semantic and lexical encoding conditions. The results revealed that both age groups exhibited robust priming. The older participants showed better performance than the young adults. Both groups exhibited ERP repetition effects at posterior sites, but only the older adults showed additional frontal activity. The results suggest that highly performing older adults compensate for their lower level of parieto-occipital functioning, reflected by smaller P300 amplitude at posterior sites, by recruiting frontal sites as a mode of brain adaptation. PMID- 19815189 TI - Perspectives of couples with high risk of transmitting genetic disorders. AB - OBJECTIVE: To investigate the preference for preimplantation genetic diagnosis (PGD) as an alternative to prenatal diagnosis (PND) in a large group of couples representing a wide array of genetic disorders. We also investigated the couple's familiarity with PGD and presented time trade-off scenarios for PGD versus PND, as PGD treatment is regularly accompanied by waiting lists. DESIGN: Questionnaire study. SETTING: Patient organizations representing genetic disorders. PATIENT(S): A total of 210 couples carrying genetic disorders. MAIN OUTCOME MEASURE(S): Preference for PGD or PND and familiarity with PGD in carrier couples. RESULT(S): Fifteen organizations representing 38 genetic disorders agreed to participate. Nine hundred eighty-three couples responded. In total 210 couples were in their reproductive years (women 18-40 years) and had a desire to conceive. Ninety couples (42%) had never heard of PGD. After they were informed, 127 couples (60%) wanted to have diagnostic testing (PND or PGD) performed. Ninety-four (74%) of these couples preferred testing with PGD. When no waiting list was used 102 couples (80%) preferred PGD. With a 2-year waiting list for PGD, 58 couples (46%) would opt for PGD. CONCLUSION(S): Many carrier couples are unaware of the existence of PGD. When informed, most couples prefer PGD more than PND. The preference for PGD decreases with longer waiting lists. PMID- 19815190 TI - Reproductive health of women electing bariatric surgery. AB - OBJECTIVE: To describe the reproductive health history and characteristics of women having bariatric surgery and to determine whether this differs by age of onset of obesity. DESIGN: Retrospective and cross-sectional analyses of self reported survey data. SETTING: Six sites of the Longitudinal Assessment of Bariatric Surgery-2 study. PATIENT(S): The study included 1,538 females having bariatric surgery. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Reported polycystic ovary syndrome (PCOS), pregnancy and fertility history, contraceptive use, and plans for pregnancies. RESULT(S): Mean age was 44.8 years (range, 18-78 years); mean body mass index was 47.2 kg/m2 (range, 33.8-87.3 kg/m2). PCOS had been diagnosed by a health care provider in 13.1% of subjects. Of women who had tried to conceive, 41.9% experienced infertility and 61.4% had a live birth after experiencing infertility. In the whole group, prior live birth was reported by 72.5%. Women who were obese by 18 years old were more likely to report PCOS and infertility and less likely to have ever been pregnant, compared with women who became obese later in life. Future pregnancy was important to 30.3% of women younger than 45 years, whereas 48.6% did not plan to become pregnant in the future. In the year before surgery, 51.8% used contraception. CONCLUSION(S): Self reporting of obesity by age 18 appears to be related to reproductive morbidity. Women undergoing bariatric surgery have important reproductive health care needs, including reliable contraception and counseling about plans for postoperative pregnancy. PMID- 19815191 TI - Defective endometrial prostaglandin synthesis identified in patients with repeated implantation failure undergoing in vitro fertilization. AB - OBJECTIVE: To define the role of prostaglandins (PG) in the endometrium of patients with repeated failure of embryo implantation. Prostaglandins are produced after the sequential oxidation of arachidonic acid by cyclooxygenases (COX-1 and COX-2) and terminal PG synthases. DESIGN: Case-control study. SETTING: In vitro fertilization unit at a university hospital. PATIENT(S): Thirty-four women, comprising of 19 patients with repeated IVF failure and 15 controls with proven fertility. INTERVENTION(S): Endometrial expression levels of the enzymes responsible for the PG synthesis were compared between the two groups. MAIN OUTCOME MEASURE(S): Cytosolic phospholipase A2 (cPLA2alpha) expression and activity were assessed by Western blot. Expression of cyclooxygenase-2, secretory phospholipase A2 group IIA, V, and IB (sPLA2-IIA, sPLA2-V, sPLA2-IB), glypican-1, PG E synthase, PG E receptors, and lysophosphatidic acid receptor 3 (LPA3) was measured by real-time polymerase chain reaction (PCR). Localization of COX-2, sPLA2-IIA, and LPA3 within the secretory endometrium was detected by immunohistochemistry. RESULT(S): Patients displaying recurrent implantation failure expressed reduced levels of cPLA2alpha and COX-2 compared with controls. In response to this deficiency, sPLA2-IIA was found to be overexpressed. Interestingly, LPA3, which is known to converge on the cPLA2-arachidonic acid-COX PG signaling pathway, was also decreased in these patients. CONCLUSION(S): Prostaglandin synthesis appears to be disrupted in patients with repeated IVF failure compared with fertile controls. We therefore suggest that reduced PG synthesis in the human endometrium may lead to poor endometrial receptivity. PMID- 19815192 TI - Comparison of pregnancy outcomes in anonymous shared versus exclusive donor oocyte in vitro fertilization cycles. AB - OBJECTIVE: To determine whether there is a difference in pregnancy outcomes between women undergoing a shared versus exclusive donor oocyte cycle. DESIGN: Retrospective analysis. SETTING: University IVF center. PATIENT(S): Women undergoing either a shared (n=656 cycles), exclusive (n=225 cycles), or shared converted to exclusive (n=22 cycles) donor oocyte cycle from January 2000 December 2005. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Number of eggs retrieved, eggs fertilized, embryos transferred, embryos cryopreserved, clinical pregnancy rates (PR), live birth rates, spontaneous abortion rates. RESULT(S): Pregnancy outcomes in 656 shared cycles were compared with 225 exclusive cycles and 22 shared converted to exclusive donor oocyte cycles. Overall, there was no difference in the clinical PR among the three groups; however, the exclusive group did have a significantly greater number of embryos cryopreserved and this event occurred more frequently in such a cycle. CONCLUSION(S): Women undergoing a donor oocyte IVF cycle can choose to share the donor's oocytes with another recipient without compromising their PR; however, the probability of cryopreservation in such a shared donor oocyte cycle is significantly reduced. Therefore, the recipient must weigh the financial burden of an exclusive cycle with the desires for cryopreservation in an IVF cycle. PMID- 19815193 TI - Increased fetal cell trafficking in murine lung following complete pregnancy loss from exposure to lipopolysaccharide. AB - To determine whether chemically induced miscarriage affects fetomaternal trafficking in a mouse model, we measured the amount of fetal DNA present in various maternal organs by polymerase chain reaction amplification following exposure to lipopolysaccharide (LPS). As the frequency of fetal cells and the number of animals with detectable microchimerism following LPS injection were significantly increased, particularly in lung tissue compared to controls, with no signs of an inflammatory response, we conclude that LPS-induced miscarriage results in increased murine fetomaternal cell trafficking, supporting a relationship between fetal loss and the establishment of fetal cell microchimerism. PMID- 19815194 TI - Outcomes of day-1, day-3, and blastocyst cryopreserved embryo transfers. AB - The optimal developmental stage for cryopreserving embryos in IVF-ET remains controversial. Our study demonstrates that besides an improvement in postthaw survival rate for day-1 and blastocyst cryopreserved ET over day-3, all three groups attained statistically similar implantation, clinical pregnancy, multiple, twinning, and male gender rates. PMID- 19815195 TI - Potential indications for ovarian autotransplantation based on the analysis of 5,571 autopsy findings of females under the age of 40 in Japan. AB - Ovarian cryopreservation and autotransplantation could be of potential value for preservation of fertility in the patients with various malignancies. Ovarian tissue should be cryopreserved actively for fertility preservation, but stored tissue should be autotransplanted with much caution until reliable methods are established to detect minimal residual disease in grafts in precise and reproducible manners. PMID- 19815196 TI - Demographic variables related to successful anonymous oocyte donor recruitment. AB - Only 4% of initial applicants to the oocyte donor program completed the screening process and actually donated a cohort of oocytes. Medical screen-outs were more likely for women who had children, were unmarried, and were less educated, whereas psychological screen-outs were more common among younger applicants, and both younger and African-American women were more likely to withdraw without donating. PMID- 19815197 TI - Factors affecting success rates in two concurrent clinical IVF trials: an examination of potential explanations for the difference in pregnancy rates between the United States and Europe. AB - OBJECTIVE: To compare a US clinical trial of gonadotropin therapy for IVF with a similar European trial to determine what factors may explain the higher clinical pregnancy rate in the US trial. DESIGN: Comparison of baseline, treatment, and outcome variables in the United States (US) and European trials. SETTING: IVF practices in the US (n=4) and Europe (n=6). PATIENT(S): 297 women undergoing IVF. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate. RESULT(S): Clinical pregnancy rates were 43.4% in the US compared with 29.7% in Europe (p=0.016), with a live birth rate of 38.2% versus 27.6% (p=0.064). This difference in clinical pregnancy rate could not be explained by differences in the US versus Europe for number of embryos transferred (2.3 vs. 2.6) or female age (34.6 vs. 30.4). Although the starting dose of gonadotropin was higher in the US trial compared with the European trial (300 versus 225 IU), the total dose of gonadotropin was only slightly higher in the US. In multiple logistic regression analysis of 81 pretransfer variables on clinical pregnancy, the only two found to be significant predictors of outcome were baseline endometrial thickness following down-regulation and number of days of gonadotropin treatment. CONCLUSION(S): This study suggests the possibility that US pregnancy rates may be higher in part because of differences in down-regulation or gonadotropin dosing. Other factors not assessed in these studies or in national datasets likely also contribute to the difference in pregnancy rates. PMID- 19815198 TI - Serum and follicular endocrine profile is different in modified natural cycles than in cycles stimulated with gonadotropin and gonadotropin-releasing hormone antagonist. AB - This study compared serum and follicular anti-mullerian hormone, LH, FSH, E(2), and P concentrations in modified natural cycles and controlled ovarian hyperstimulation cycles to analyze their effect on oocyte and embryo quality and implantation. Follicular anti-mullerian hormone, FSH, E(2), and P concentrations had no effect on embryo quality, whereas high serum and follicular LH concentrations in natural cycles modified only with hCG did not compromise oocyte and embryo quality but might have a negative effect on implantation. PMID- 19815199 TI - Dynamic simulation and nonlinear control of a rigorous batch reactive distillation. AB - This work deals with the dynamics and control of a high-purity batch distillation column with chemical reaction. A heterogeneous esterification reaction between the acetic acid and butanol takes place to produce butyl acetate. The process model is formulated considering variable liquid holdup, UNIQUAC model for thermodynamic property predictions, nonlinear Francis weir formula for tray hydraulics, pseudohomogeneous model to represent the reaction kinetics and rigorous energy balance. A structured and simple iterative approach is devised to compute the vapor flows with the fast convergence, under the rigorous energy balance. The representative column is treated with a distillate policy based on which, the lightest product, water is removed as distillate at the starting of production phase. As a consequence, the column gets progressively richer with the main product, butyl acetate. In addition, almost complete conversion of the limiting reactant is achieved. In order to maintain the product purity at the top, a nonlinear generic model controller (GMC) in two different forms has been proposed. Finally, a comparative closed-loop performance is addressed. It is shown that the control scheme, along with the effective distillate strategy, leads to almost complete conversion of ingredients and high-purity products. PMID- 19815200 TI - 60 Million non-facility births: who can deliver in community settings to reduce intrapartum-related deaths? AB - BACKGROUND: For the world's 60 million non-facility births, addressing who is currently attending these births and what effect they have on birth outcomes is a key starting point toward improving care during childbirth. OBJECTIVE: We present a systematic review of evidence for the effect of community-based cadres community-based skilled birth attendants (SBAs), trained traditional birth attendants (TBAs), and community health workers (CHWs)-in improving perinatal and intrapartum-related outcomes. RESULTS: The evidence for providing skilled birth attendance in the community is low quality, consisting of primarily before-and after and quasi-experimental studies, with a pooled 12% reduction in all cause perinatal mortality (PMR) and a 22%-47% reduction in intrapartum-related neonatal mortality (IPR-NMR). Low/moderate quality evidence suggests that TBA training may improve linkages with facilities and improve perinatal outcomes. A randomized controlled trial (RCT) of TBA training showed a 30% reduction in PMR, and a meta analysis demonstrated an 11% reduction in IPR-NMR. There is moderate evidence that CHWs have a positive impact on perinatal-neonatal outcomes. Meta-analysis of CHW packages (2 cluster randomized controlled trials, 2 quasi-experimental studies) showed a 28% reduction in PMR and a 36% reduction in early neonatal mortality rate; one quasi-experimental study showed a 42% reduction in IPR-NMR. CONCLUSION: Skilled childbirth care is recommended for all pregnant women, and community strategies need to be linked to prompt, high-quality emergency obstetric care. CHWs may play a promising role in providing pregnancy and childbirth care, mobilizing communities, and improving perinatal outcomes in low income settings. While the role of the TBA is still controversial, strategies emphasizing partnerships with the health system should be further considered. Innovative community-based strategies combined with health systems strengthening may improve childbirth care for the rural poor, help reduce gross inequities in maternal and newborn survival and stillbirth rates, and provide an effective transition to higher coverage for facility births. PMID- 19815202 TI - Two million intrapartum-related stillbirths and neonatal deaths: where, why, and what can be done? AB - BACKGROUND: Intrapartum-related neonatal deaths ("birth asphyxia") are a leading cause of child mortality globally, outnumbering deaths from malaria. Reduction is crucial to meeting the fourth Millennium Development Goal (MDG), and is intimately linked to intrapartum stillbirths as well as maternal health and MDG 5, yet there is a lack of consensus on what works, especially in weak health systems. OBJECTIVE: To clarify terminology for intrapartum-related outcomes; to describe the intrapartum-related global burden; to present current coverage and trends for care at birth; and to outline aims and methods for this comprehensive 7-paper supplement reviewing strategies to reduce intrapartum-related deaths. RESULTS: Birth is a critical time for the mother and fetus with an estimated 1.02 million intrapartum stillbirths, 904,000 intrapartum-related neonatal deaths, and around 42% of the 535,900 maternal deaths each year. Most of the burden (99%) occurs in low- and middle-income countries. Intrapartum-related neonatal mortality rates are 25-fold higher in the lowest income countries and intrapartum stillbirth rates are up to 50-fold higher. Maternal risk factors and delays in accessing care are critical contributors. The rural poor are at particular risk, and also have the lowest coverage of skilled care at birth. Almost 30,000 abstracts were searched and the evidence is evaluated and reported in the 6 subsequent papers. CONCLUSION: Each year the deaths of 2 million babies are linked to complications during birth and the burden is inequitably carried by the poor. Evidence-based strategies are urgently needed to reduce the burden of intrapartum-related deaths particularly in low- and middle-income settings where 60 million women give birth at home. PMID- 19815201 TI - Linking families and facilities for care at birth: what works to avert intrapartum-related deaths? AB - BACKGROUND: Delays in receiving effective care during labor and at birth may be fatal for the mother and fetus, contributing to 2 million annual intrapartum stillbirths and intrapartum-related neonatal deaths each year. OBJECTIVE: We present a systematic review of strategies to link families and facilities, including community mobilization, financial incentives, emergency referral and transport systems, prenatal risk screening, and maternity waiting homes. RESULTS: There is moderate quality evidence that community mobilization with high levels of community engagement can increase institutional births and significantly reduce perinatal and early neonatal mortality. Meta-analysis showed a doubling of skilled birth attendance and a 36% reduction in early neonatal mortality. However, no data are available on intrapartum-specific outcomes. Evidence is limited, but promising, that financial incentive schemes and community referral/transport systems may increase rates of skilled birth attendance and emergency obstetric care utilization; however, impact on mortality is unknown. Current evidence for maternity waiting homes and risk screening is low quality. CONCLUSIONS: Empowering communities is an important strategy to reduce the large burden of intrapartum complications. Innovations are needed to bring the poor closer to obstetric care, such as financial incentives and cell phone technology. New questions need to be asked of "old" strategies such as risk screening and maternity waiting homes. The effect of all of these strategies on maternal and perinatal mortality, particularly intrapartum-related outcomes, requires further evaluation. PMID- 19815203 TI - Neonatal resuscitation in low-resource settings: what, who, and how to overcome challenges to scale up? AB - BACKGROUND: Each year approximately 10 million babies do not breathe immediately at birth, of which about 6 million require basic neonatal resuscitation. The major burden is in low-income settings, where health system capacity to provide neonatal resuscitation is inadequate. OBJECTIVE: To systematically review the evidence for neonatal resuscitation content, training and competency, equipment and supplies, cost, and key program considerations, specifically for resource constrained settings. RESULTS: Evidence from several observational studies shows that facility-based basic neonatal resuscitation may avert 30% of intrapartum related neonatal deaths. Very few babies require advanced resuscitation (endotracheal intubation and drugs) and these newborns may not survive without ongoing ventilation; hence, advanced neonatal resuscitation is not a priority in settings without neonatal intensive care. Of the 60 million nonfacility births, most do not have access to resuscitation. Several trials have shown that a range of community health workers can perform neonatal resuscitation with an estimated effect of a 20% reduction in intrapartum-related neonatal deaths, based on expert opinion. Case studies illustrate key considerations for scale up. CONCLUSION: Basic resuscitation would substantially reduce intrapartum-related neonatal deaths. Where births occur in facilities, it is a priority to ensure that all birth attendants are competent in resuscitation. Strategies to address the gap for home births are urgently required. More data are required to determine the impact of neonatal resuscitation, particularly on long-term outcomes in low income settings. PMID- 19815204 TI - Obstetric care in low-resource settings: what, who, and how to overcome challenges to scale up? AB - BACKGROUND: Each year, approximately 2 million babies die because of complications of childbirth, primarily in settings where effective care at birth, particularly prompt cesarean delivery, is unavailable. OBJECTIVE: We reviewed the content, impact, risk-benefit, and feasibility of interventions for obstetric complications with high population attributable risk of intrapartum-related hypoxic injury, as well as human resource, skill development, and technological innovations to improve obstetric care quality and availability. RESULTS: Despite ecological associations of obstetric care with improved perinatal outcomes, there is limited evidence that intrapartum interventions reduce intrapartum-related neonatal mortality or morbidity. No interventions had high-quality evidence of impact on intrapartum-related outcomes in low-resource settings. While data from high-resource settings support planned cesarean for breech presentation and post term induction, these interventions may be unavailable or less safe in low resource settings and require risk-benefit assessment. Promising interventions include use of the partograph, symphysiotomy, amnioinfusion, therapeutic maneuvers for shoulder dystocia, improved management of intra-amniotic infections, and continuous labor support. Obstetric drills, checklists, and innovative low-cost devices could improve care quality. Task-shifting to alternative cadres may increase coverage of care. CONCLUSIONS: While intrapartum care aims to avert intrapartum-related hypoxic injury, rigorous evidence is lacking, especially in the settings where most deaths occur. Effective care at birth could save hundreds of thousands of lives a year, with investment in health infrastructure, personnel, and research--both for innovation and to improve implementation. PMID- 19815205 TI - Reducing intrapartum-related deaths and disability: can the health system deliver? AB - BACKGROUND: Each year 1.02 million intrapartum stillbirths and 904,000 intrapartum-related neonatal deaths (formerly called "birth asphyxia") occur, closely linked to 536,000 maternal deaths, an estimated 42% of which are intrapartum-related. OBJECTIVE: To summarize the results of a systematic evidence review, and synthesize actions required to strengthen healthcare delivery systems and home care to reduce intrapartum-related deaths. METHODS: For this series, systematic searches were undertaken, data synthesized, and meta-analyses carried out for various aspects of intrapartum care, including: obstetric care, neonatal resuscitation, strategies to link communities with facility-based care, care within communities for 60 million non-facility births, and perinatal audit. We used the Lives Saved Tool (LiST) to estimate neonatal deaths prevented with relevant interventions under 2 scenarios: (1) to address missed opportunities for facility and home births; and (2) assuming full coverage of comprehensive emergency obstetric care and emergency newborn care. Countries were first grouped into 5 Categories according to level of neonatal mortality rate and examined, and then priorities were suggested to reduce intrapartum-related deaths for each Category based on health performance and possible lives saved. RESULTS: There is moderate GRADE evidence of effectiveness for the reduction of intrapartum-related mortality through facility-based neonatal resuscitation, perinatal audit, integrated community health worker packages, and community mobilization. The quality of evidence for obstetric care is low, requiring further evaluation for effect on perinatal outcomes, but is expected to be high impact. Over three quarters of intrapartum-related deaths occur in settings with weak health systems marked by low coverage of skilled birth attendance (<50%), low density of skilled human resources (<0.9 per 1000 population) and low per capita spending on health (9km/day run produced risks 65% lower for angina (P=0.008), 29% lower for nonfatal CHD (P=0.04), and 26% lower for fatal and nonfatal CHD (P=0.06). CONCLUSIONS: Exceeding guideline physical activity levels produce important CHD risk reductions. PMID- 19815209 TI - A role for MMP-3 genetic variation in atherosclerosis susceptibility? PMID- 19815210 TI - NT-proBNP is associated with coronary heart disease risk in healthy older women but fails to enhance prediction beyond established risk factors: results from the British Women's Heart and Health Study. AB - OBJECTIVE: Limited evidence suggests NT-proBNP improves prediction of coronary heart disease (CHD) events but further data are needed, especially in people without pre-existing CHD and in women. METHODS: We measured NT-proBNP in serum from 162 women with incident CHD events and 1226 controls (60-79 years) in a case control study nested within the prospective British Women's Heart and Health Study. All cases and controls were free from CHD at baseline. We related NT proBNP to CHD event risk, and determined to what extent NT-proBNP enhanced CHD risk prediction beyond established risk factors. RESULTS: The odds ratio for CHD per 1 standard deviation increase in log(e)NT-proBNP was 1.37 (95% CI: 1.13-1.68) in analyses adjusted for established CHD risk factors, social class, CRP and insulin. However, addition of log(e)NT-proBNP did not improve the discrimination of a prediction model including age, social class, smoking, physical activity, lipids, fasting glucose, waist:hip ratio, hypertension, statin and aspirin use, nor a standard Framingham risk score model; area under the receiver operator curve for the former model increased from 0.676 to 0.687 on inclusion of NT proBNP (p=0.3). Furthermore, adding NT-proBNP did not improve calibration of a prediction model containing established risk factors, nor did inclusion more appropriately re-classify participants in relation to their final outcome. Findings were similar (independent associations, but no prediction improvement) for fasting insulin and CRP. CONCLUSION: These results caution against use of NT proBNP for CHD risk prediction in healthy women and suggest a need for larger studies in both genders to resolve outstanding uncertainties. PMID- 19815211 TI - Mandibular bone remodeling induced by dental implant. AB - The ability to assess the effects of an implant on bone remodeling is of particular importance to prosthesis placement planning and associated treatment assurance. Prediction of on-going bone responses will enable us to improve the performance of a restoration. Although the bone remodeling for long bones had been extensively studied, there have been relatively few reports for dental scenarios despite its increasing significance with more and more dental implant placements. This paper aimed to develop a systematic protocol to assess mandibular bone remodeling induced by dental implantation, which extends the remodeling algorithms established for the long bones into dental settings. In this study, a 3D model for a segment of a human mandible was generated from in vivo CT scan images, together with a titanium implant embedded to the mandible. The results examined the changes in bone density and stiffness as a result of bone remodeling over a period of 48 months. Resonance frequency analysis was also performed to relate natural frequencies to bone remodeling. The density contours are qualitatively compared with clinical follow-up X-ray images, thereby providing validity for the bone remodeling algorithm presented in dental bone analysis. PMID- 19815212 TI - Innovations in cell mechanobiology. PMID- 19815213 TI - Emergent morphogenesis: elastic mechanics of a self-deforming tissue. AB - Multicellular organisms are generated by coordinated cell movements during morphogenesis. Convergent extension is a key tissue movement that organizes mesoderm, ectoderm, and endoderm in vertebrate embryos. The goals of researchers studying convergent extension, and morphogenesis in general, include understanding the molecular pathways that control cell identity, establish fields of cell types, and regulate cell behaviors. Cell identity, the size and boundaries of tissues, and the behaviors exhibited by those cells shape the developing embryo; however, there is a fundamental gap between understanding the molecular pathways that control processes within single cells and understanding how cells work together to assemble multicellular structures. Theoretical and experimental biomechanics of embryonic tissues are increasingly being used to bridge that gap. The efforts to map molecular pathways and the mechanical processes underlying morphogenesis are crucial to understanding: (1) the source of birth defects, (2) the formation of tumors and progression of cancer, and (3) basic principles of tissue engineering. In this paper, we first review the process of tissue convergent extension of the vertebrate axis and then review models used to study the self-organizing movements from a mechanical perspective. We conclude by presenting a relatively simple "wedge-model" that exhibits key emergent properties of convergent extension such as the coupling between tissue stiffness, cell intercalation forces, and tissue elongation forces. PMID- 19815215 TI - Exogenous and endogenous force regulation of endothelial cell behavior. AB - Endothelial cells live in a dynamic environment where they are constantly exposed to external hemodynamic forces and generate cytoskeletal-based endogenous forces. These exogenous and endogenous forces are critical regulators of endothelial cell health and blood vessel maintenance at all generations of the vascular system, from large arteries to capillary beds. The first part of this review highlights the role of the primary exogenous hemodynamic forces of shear, cyclic strain, and pressure forces in mediating endothelial cell response. We then discuss the emergent role of the mechanical properties of the extracellular matrix and of cellular endogenous force generation on endothelial cell function, implicating substrate stiffness and cellular traction stresses as important mediators of endothelial cell health. The intersection of exogenous and endogenous forces on endothelial cell function is discussed, suggesting some of the many remaining questions in the field of endothelial mechanobiology. PMID- 19815214 TI - Shape, loading, and motion in the bioengineering design, fabrication, and testing of personalized synovial joints. AB - With continued development and improvement of tissue engineering therapies for small articular lesions, increased attention is being focused on the challenge of engineering partial or whole synovial joints. Joint-scale constructs could have applications in the treatment of large areas of articular damage or in biological arthroplasty of severely degenerate joints. This review considers the roles of shape, loading and motion in synovial joint mechanobiology and their incorporation into the design, fabrication, and testing of engineered partial or whole joints. Incidence of degeneration, degree of impairment, and efficacy of current treatments are critical factors in choosing a target for joint bioengineering. The form and function of native joints may guide the design of engineered joint-scale constructs with respect to size, shape, and maturity. Fabrication challenges for joint-scale engineering include controlling chemo mechano-biological microenvironments to promote the development and growth of multiple tissues with integrated interfaces or lubricated surfaces into anatomical shapes, and developing joint-scale bioreactors which nurture and stimulate the tissue with loading and motion. Finally, evaluation of load-bearing and tribological properties can range from tissue to joint scale and can focus on biological structure at present or after adaptation. PMID- 19815216 TI - Musculoskeletal mechanobiology: interpretation by external force and engineered substratum. AB - Mechanobiology aims to discover how the mechanical environment affects the biological activity of cells and how cells' ability to sense these mechanical cues is converted into elicited cellular responses. Musculoskeletal mechanobiology is of particular interest given the high mechanical loads that musculoskeletal tissues experience on a daily basis. How do cells within these mechanically active tissues interpret external loads imposed on their extracellular environment, and, how are cell-substrate interactions converted into biochemical signals? This review outlines many of the main mechanotransduction mechanisms known to date, and describes recent literature examining effects of both external forces and cell-substrate interactions on musculoskeletal cells. Whether via application of external forces and/or cell substrate interactions, our understanding and regulation of musculoskeletal mechanobiology can benefit by expanding upon traditional models, and shedding new light through novel investigative approaches. Current and future work in this field is focused on identifying specific forces, stresses, and strains at the cellular and tissue level through both experimental and computational approaches, and analyzing the role of specific proteins through fluorescence-based investigations and knockdown models. PMID- 19815217 TI - Mechanics of microtubules. AB - Microtubules are rigid cytoskeletal filaments, and their mechanics affect cell morphology and cellular processes. For instance, microtubules for the support structures for extended morphologies, such as axons and cilia. Further, microtubules act as tension rods to pull apart chromosomes during cellular division. Unlike other cytoskeletal filaments (e.g., actin) that work as large networks, microtubules work individually or in small groups, so their individual mechanical properties are quite important to their cellular function. In this review, we explore the past work on the mechanics of individual microtubules, which have been studied for over a quarter of a century. We also present some prospective on future endeavors to determine the molecular mechanisms that control microtubule rigidity. PMID- 19815218 TI - Expansion and hydrodynamic properties of beta-cyclodextrin polymer/tungsten carbide composite matrix in an expanded bed. AB - The expansion and hydrodynamic properties of matrix are significant for expanded bed adsorption (EBA) processes. A series of new composite matrices CroCD-TuC are studied and estimated in an expanded bed. It is found that the heavier matrix is better suited for high operation fluid velocity than the lighters. Although the Richardson-Zaki equation can well correlate the bed voidage with fluid velocity for all CroCD-TuC matrices tested, the modifications are proposed to improve the accuracy of theoretical predictions of correlation parameters, including terminal settling velocity (U(t)) and expansion index (n). Residence time distributions (RTDs) are determined, and the Bodenstein number (Bo) and axial dispersion coefficient (D(ax)) are employed to analyze the liquid mixing in the expanded bed. It is found for CroCD-TuC matrices, both parameters notably changed with the variation of fluid velocity and viscosity. Furthermore, D(ax) is an intuitive parameter estimating the bed stability on various operating conditions, and also a restriction on developing the matrix for high operation fluid velocity. The comparison of the hydrodynamic properties on different matrices reveals that CroCD-TuC 3 and CroCD-TuC 4 seem superior to other matrices in hydrodynamic properties, making them promising matrices for further use. The correlations as the functions of fluid velocity and viscosity have been established which may provide beneficial information for practical applications of CroCD-TuC matrices in EBA processes. PMID- 19815219 TI - A microscale approach for predicting the performance of chromatography columns used to recover therapeutic polyclonal antibodies. AB - A microscale approach is described which screens conditions for recovering polyclonal antibodies from ovine sera by mixed-mode cation-exchange chromatography. The impact of pH and loading buffer salt concentration were assessed using robotically operated 20microL packed pipette tips. Low salt concentrations delivered capacities up to 41mg/mL, while only half this level was obtained at high salt concentrations. Two of the screened conditions were then tested in a 10mL packed bed and overall trends in capacity, yield and purity were found to be retained. Microscale pipette tips thus provided a useful basis for the rapid, approximate definition of a chromatography design space. PMID- 19815220 TI - Engineering of a two-step purification strategy for a panel of monoclonal immunoglobulin M directed against undifferentiated human embryonic stem cells. AB - A two-step purification strategy comprising of polyethylene glycol (PEG) precipitation and anion-exchange chromatography was developed for a panel of monoclonal immunoglobulin M (IgM) (pI 5.5-7.7) produced from hybridoma cultures. PEG precipitation was optimized with regards to concentration, pH and mixing. For anion-exchange chromatography, different resins were screened of which Fractogel EMD, a polymer grafted porous resin had the highest capacity. Despite its significantly slower mass transfer, the binding capacity was still higher compared to a convection driven resin (monolith). This purification strategy was successfully demonstrated for all 9 IgMs in the panel. In small scale most antibodies could be purified to >95% purity with the exception of two which gave a lower final purity (46% and 85%). The yield was dependent on the different antibodies ranging from 28% to 84%. Further improvement of recovery and purity was obtained by the digestion of DNA present in the hybridoma supernatant using an endonuclease, benzonase. So far this strategy has been applied for the purification of up to 2l hybridoma supernatants. PMID- 19815221 TI - Rapid high performance liquid chromatography method development with high prediction accuracy, using 5cm long narrow bore columns packed with sub-2microm particles and Design Space computer modeling. AB - Many different strategies of reversed phase high performance liquid chromatographic (RP-HPLC) method development are used today. This paper describes a strategy for the systematic development of ultrahigh-pressure liquid chromatographic (UHPLC or UPLC) methods using 5cmx2.1mm columns packed with sub 2microm particles and computer simulation (DryLab((R)) package). Data for the accuracy of computer modeling in the Design Space under ultrahigh-pressure conditions are reported. An acceptable accuracy for these predictions of the computer models is presented. This work illustrates a method development strategy, focusing on time reduction up to a factor 3-5, compared to the conventional HPLC method development and exhibits parts of the Design Space elaboration as requested by the FDA and ICH Q8R1. Furthermore this paper demonstrates the accuracy of retention time prediction at elevated pressure (enhanced flow-rate) and shows that the computer-assisted simulation can be applied with sufficient precision for UHPLC applications (p>400bar). Examples of fast and effective method development in pharmaceutical analysis, both for gradient and isocratic separations are presented. PMID- 19815222 TI - Binding and elution behavior of proteins on strong cation exchangers. AB - This work provides a broad survey of binding and elution behavior of proteins on strong cation exchangers. Four proteins comprising two monoclonal antibodies, lysozyme, and cytochrome c were used as models in the investigation. Seven chromatography resins with different base matrices were compared. Dynamic binding capacity as a function of salt concentration was examined for a monoclonal antibody and lysozyme. Elution behavior as a function of gradient slope was modeled to determine the characteristic charge, essentially a measure of the number of sites involved in binding, for each protein on each resin. Trends with respect to dynamic binding capacity and elution behavior are analyzed and discussed. PMID- 19815223 TI - Group-specific fragmentation of pesticides and related compounds in liquid chromatography-tandem mass spectrometry. AB - Current strategies in the LC-MS analysis of pesticides and related compounds in environmental samples, fruits and vegetables, and biological samples mostly rely on the selection of appropriate precursor/product-ion combinations (transitions) for selected reaction monitoring (SRM), often based on automated parameter optimization and selection of the transition. Such a procedure does not require any information on the type of fragmentation reaction involved in the generation of the product ion from the selected precursor ion. However, such information does become important in untargeted screening for unknown contaminants in environmental and food samples, which are generally based on a combination of high-resolution mass spectrometry and (multistage) tandem mass spectrometry. With this in mind, the group-specific fragmentation behaviour has been studied for six classes of pesticides and herbicides, i.e., triazines, organophosphorous pesticides, phenylurea herbicides, carbamates, sulfonylurea herbicides, and chlorinated phenoxy acid herbicides. When relevant, some comparison was made between fragmentation of protonated molecules in MS-MS and of molecular ions generated by electron ionization in GC-MS. PMID- 19815224 TI - Analysis of perfluorinated chemicals in sludge: method development and initial results. AB - A fast, rigorous method was developed to maximize the extraction efficacy for ten perfluorocarboxylic acids and perfluorooctanesulfonate from waste-water-treatment sludge and to quantitate using liquid chromatography, tandem-mass spectrometry (LC/MS/MS). First, organic solvents were tested for extraction efficiency, including acetonitrile (ACN), methanol (MeOH), isopropanol (IPA), tetrahydrofuran (THF), and 50/50 ACN/MeOH (v/v). Among the extractants tested, 50/50 ACN/MeOH yielded the best results for our combined criteria of extraction efficacy and solvent-handling convenience. Second, chemical pretreatment prior to solvent extraction was tested with sodium hydroxide (NaOH), potassium hydroxide (KOH), hydrochloric acid (HCl), and potassium persulfate (K(2)S(2)O(8)). Pretreatment with NaOH and HCl effectively recovered additional PFCs from the sludge, but KOH and K(2)S(2)O(8) digestion were less effective than no pretreatment. Third, cleanup methods were investigated with solid-phase extraction using HLB (hydrophilic-lipophilic balanced) and WAX (weak-anion exchange) stationary phases, and with ion-pairing. The HLB stationary phase yielded a slight edge over the other two cleanup strategies in terms of recoverable PFCs and chromatographic separation. Finally, the appropriateness of isotopically labeled PFCs for quantitating unlabeled PFCs using isotopic dilution in complex sludge extracts was evaluated by comparison to results obtained with the standard-addition method. A National Institute of Standards and Technology (NIST) domestic sludge (CRM 2781) was analyzed using our finalized method and compared with previously reported results. PMID- 19815225 TI - Enantiomeric resolution of biomarkers in space analysis: Chemical derivatization and signal processing for gas chromatography-mass spectrometry analysis of chiral amino acids. AB - The work compares two GC-MS methods for enantioselective separation of amino acids as suitable candidate for stereochemical analysis of chiral amino acids on board spacecrafts in space exploration missions of solar system body environments. Different derivatization reagents are used: a mixture of alkyl chloroformate-alcohol-pyridine to obtain the alkyl alkoxy carbonyl esters and a mixture of perfluorinated alcohols and anhydrides to form perfluoroacyl perfluoroalkyl esters. 20 proteinogenic amino acids were derivatized with the two procedures and submitted to GC-MS analysis on a Chirasil-l-Val stationary phase. The results were then compared in terms of the enantiomeric separation achieved and intensity of MS response. The combination of methyl chloroformate (MCF) and heptafluoro-1-butanol (HFB) allows separation of 14 enantiomeric pairs, five of which display a resolution (R(s)>or=1.2) supposed to be sufficient to quantify the enantiomeric excess. Three mixtures of trifluoroacetic (TFAA) and heptafluorobutyric (HFBA) anhydrides were combined with the corresponding perfluorinated alcohols - TFE (2,2,2-trifluoro-1-ethanol) and HFB (2,2,3,3,4,4,4 heptafluoro-1-butanol) - to give three different reagents (TFAA-TFE, TFAA-HFB, HFBA-HFB): the derivatives obtained show separation of the same number of proteinogenic amino acids (14 of 20) at a temperature lower than column bleeding limit (200 degrees C) and 8 of them give a separation with R(s)>or=1.2. Linearity study and limit of detection (X(LOD)) computation show that both methods are suitable for quantitative determination of several amino acid diastereomers at trace level (X(LOD) approximately 0.5nmol as derivatized quantity). Both the procedures were coupled with automatic data handling to increase their suitability for space analysis: the simplified data treatment is especially helpful to handle the low quality data recovered from space experiments and labor and time are saved, as imposed by the space experiments requiring a rapid delivery of the results. To achieve this aim, a chemometric approach based on the computation of the Autocovariance Function (ACVF) was applied to extract information on the enantiomeric pairs present in the sample and the enantioseparation achieved on the chiral column. PMID- 19815226 TI - A thermodynamic solution model for calcium carbonate: Towards an understanding of multi-equilibria precipitation pathways. AB - Thermodynamic solubility calculations are normally only related to thermodynamic equilibria in solution. In this paper, we extend the use of such solubility calculations to help elucidate possible precipitation reaction pathways during the entire reaction. We also estimate the interfacial energy of particles using only solubility data by a modification of Mersmann's approach. We have carried this out by considering precipitation reactions as a succession of small quasi equilibrium states. Thus possible equilibrium precipitation pathways can be evaluated by calculating the evolution of surface charge, particle size and/or interfacial energy during the ongoing reaction. The approach includes the use of the Kelvin's law to express the influence of particle size on the solubility constant of precipitates, the use of Nernst's law to calculate surface potentials from solubility calculations and relate this to experimentally measured zeta potentials. Calcium carbonate precipitation and zeta potential measurements of well characterised high purity calcite have been used as a model system to validate the calculated values. The clarification of the change in zeta potential on titration illustrates the power of this approach as a tool for reaction pathway prediction and hence knowledge based tailoring of precipitation reactions. PMID- 19815227 TI - Monomeric and dimeric anionic surfactants: A comparative study of self aggregation and mineralization. AB - A study on the phase behavior and structure of the alkanolamine salts of the dimeric amphiphile 3,4-bis-dodecyloxycarbonyl-hexanedioic acid (GS-H) is presented for the first time. Data are compared to those of the corresponding monomeric surfactant (lauric acid, LA). The alkanolamine salts of GS-H show very low Krafft points (<0 degrees C) and form hexagonal liquid crystals at concentrations lower than its monomeric counterpart, indicating that aggregation is favored for dimeric surfactants. The minimum concentration for liquid crystal formation increases for bulky alkanolamines with a structure-disrupting effect, such as triethanolamine (TEA). However, the specific surface areas per molecule in the liquid crystals derived from small-angle X-ray scattering (SAXS) are similar for monoethanolamine (MEA) and TEA salts; the same can be said when comparing monomeric (LA) and dimeric (GS-H) salts. GS-H can also form hexagonal and lamellar liquid crystals with organic aminosilanes acting as reactive counterions, as revealed by solvent penetration experiments with polarized optical microscopy (POM). Consequently, mineralization with silica and alumina was carried out by a sol-gel method using GS-H as a possible structure-directing agent. Both silica and alumina samples possessed a lamellar structure, which disappears on calcination; however, calcined alumina has indeed a high surface area coming mainly from micropores. It was found that the surfactant/aminosilane ratio is critical for obtaining structured silica before calcination. PMID- 19815228 TI - Synthesis and properties of cationic surfactants with tuned hydrophylicity. AB - A series of pyridinium-based cationic surfactants has been synthesised and their amphiphilic properties have been studied by conductivity and surface tension measurements. The modification of the substitution pattern on the pyridinium ring by hydrophobic moieties (methyl vs. hydrogen and presence or not of condensed benzene ring) gave the opportunity to investigate structure-activity relationships. Characterization by conductivity and surface tension measurements shed light on the behaviour at the air/water interface and in the micellar environment. In particular, the tendency to form ion pairs at very low concentration was evidenced for all the surfactants substituted on the ring, but not for the simple pyridinium ones. The formation of ion pairs affects both the conductivity and the surface tension plots, showing that a series of steps is involved during the adsorption to the air/water surface. An attempt was made to qualify the single steps in the adsorption at the surface layer. Those steps were attributed to different chemical species (free surfactant ions or ion pairs) and to different arrangements of the surfactant. This work also represents a contribution of investigation at very low surfactant concentrations and high surface tension values. PMID- 19815229 TI - Immunohistochemical characterization of inflammatory and glial responses in a case of necrotizing leucoencephalitis in a French bulldog. AB - A 3-year-old male French bulldog was presented with blindness, staggering and ataxia and was humanely destroyed due to worsening of the neurological signs. At post-mortem examination a non-suppurative leucoencephalitis with extensive malacia within the forebrain was found. In addition, a bilateral necrotizing optic neuritis and focal retinitis was detected. Immunohistochemistry revealed a CD3(+) T-cell dominated inflammatory response with intralesional reactive astrocytes expressing glial fibrillary acidic protein. Astroglia-like cells expressing vimentin, which is characteristic of immature astrocytes, were found within the malacic lesions. The pathological findings are similar to those described in idiopathic necrotizing leucoencephalitis (NLE) of Yorkshire terriers and substantiate the hypothesis that NLE is not a breed-specific disorder that exclusively affects Yorkshire terriers, but also the French bulldog. PMID- 19815230 TI - Correlation of ST-segment "hump sign" during exercise testing with impaired diastolic function of the left ventricle. AB - BACKGROUND: The appearance of a discrete upward deflection of the ST segment, termed the ST hump sign during exercise testing has been associated with resting hypertension and exaggerated blood pressure response to exercise. OBJECTIVE: We investigated the correlation between the presence of hump sign during exercise testing with coexisting impaired diastolic function of the left ventricle (LV) at these patients. METHODS: We formed a cohort of 237 nonconsecutive patients (140 males, 41 +/- 5 years old) having undergone a treadmill test, according to the Bruce protocol, which divided into 2 groups: group A, including 130 patients which presented ST-segment hump sign at any of the leads of the electrocardiograms recorded during exercise, and group B, including 107 patients that didn't. All patients subsequently underwent an echocardiographic estimation of the LV diastolic function, using conventional and Tissue Doppler Imaging techniques. RESULTS: From 237 patients included in our study, 106 had echocardiographic signs of diastolic LV dysfunction. Among them, the appearance of ST hump sign at the peak of exercise testing was observed in 93 patients (88%), particularly in the inferior and lateral leads, while no ST hump sign was observed only in 13 patients (12%) with impaired diastolic LV function. CONCLUSIONS: The appearance of ST segment hump sign during exercise testing is strongly correlated with diastolic LV dysfunction and can be used as an exercise electrocardiographic index of diastolic LV dysfunction, independently from the echocardiographic study. PMID- 19815231 TI - ST-segment depression in aVR as a predictor of culprit artery and infarct size in acute inferior wall ST-segment elevation myocardial infarction. AB - BACKGROUND: ST-segment depression in lead aVR in acute inferior wall ST-segment elevation myocardial infarction (STEMI) has recently been suggested as a predictor of left circumflex (LCx) artery involvement. The purpose of this study is to evaluate the clinical significance of aVR depression during inferior wall STEMI. METHODS: This study included 106 consecutive patients who presented with inferior wall STEMI and underwent urgent coronary angiogram. Clinical and angiographic findings were compared between patients with and without aVR depression > or = 0.1 mV. RESULTS: The sensitivity and specificity of aVR depression as a predictor of LCx infarction were 53% and 86%, respectively. In patients with right coronary artery infarction, aVR depression was associated with increased cardiac enzymes and the involvement of a large posterolateral branch, which may explain the larger infarction. CONCLUSIONS: ST-segment depression in lead aVR in inferior wall STEMI predicts LCx infarction or larger RCA infarction involving a large posterolateral branch. PMID- 19815232 TI - Allograft inflammatory factor-1 is up-regulated in warm and cold ischemia reperfusion injury in rat liver and may be inhibited by FK506. AB - BACKGROUND: Allograft inflammatory factor-1 (AIF-1) plays an important role in immune response and vasculopathy in allografts. The present study investigated activation of AIF-1 in warm and cold ischemia-reperfusion (IR) injury of rat liver, and the potential inhibitory effect of FK506. METHODS: We used warm IR injury, orthotopic transplantation, and allograft rejection models in this study. We assessed expression of AIF-1 mRNA and protein, as well as its inducers interferon-gamma (IFN-gamma) and interleukin-1beta (IL-1beta). The potential inhibitory effect of FK506 on AIF-1 in a rat macrophage cell line and in these three models was also assessed. RESULTS: AIF-1 mRNA and protein, as well as its inducers IFN-gamma and IL-1beta, were significantly increased in the warm IR injury, orthotopic transplantation, and allograft rejection models. Real-time RT PCR and Western blotting showed that pretreatment with low-dose FK506 partially inhibited AIF-1 activation as well as its inducers (IFN-gamma and IL-1beta) in these three models. Western blotting showed that IFN-gamma and IL-1beta activated AIF-1 in a macrophage cell line, but pretreatment with FK506 did not inhibit AIF 1 activation in vitro. Hematoxylin and eosin staining showed that edema and necrosis in the liver, as well as alanine aminotransferase (ALT) in serum, of these three groups was reduced after FK506 pretreatment. CONCLUSION: AIF-1 was activated in warm and cold IR injury, and pretreatment with low-dose FK506 partly inhibited AIF-1 activation and reduced warm and cold IR injury. PMID- 19815233 TI - Change in brain glucose after enteral nutrition in subarachnoid hemorrhage. AB - BACKGROUND: The alteration of brain extracellular glucose after enteral nutrition (EN) remains unclear. In this study, we used brain microdialysis methods to estimate whether the physiologic elevation of plasma glucose following EN affects brain glucose metabolism of aneurysmal subarachnoid hemorrhage (SAH) patients. METHODS: Brain extracellular glucose, lactate, glycerol, glutamate, and pyruvate were measured with a brain microdialysis probe in 12 patients (mean age: 60.0 y+/ 7.8 y) after SAH. The EN was initially administered a mean of 3.2 d after the onset of SAH. All of the measured parameters were estimated before and after EN. RESULTS: Cerebral perfusion pressure did not significantly change after SAH during the study period. Plasma glucose rose significantly after EN (141.4+/ 11.6mg/dL before EN versus 183.8+/-26.2mg/dL immediately after EN (P=0.0006), 177.7+/-30.2mg/dL at 2h after EN (P=0.0033)). The brain extracellular glucose before EN (2.5+/-0.92mmol/L) was significantly lower than the levels measured just after (3.49+/-1.0mmol/L, P=0.0186) and 2h after the end of EN (3.70+/ 1.0mmol/L, P=0.0053). Brain extracellular concentrations of lactate, glutamate, pyruvate, and glycerol showed no significant changes. CONCLUSIONS: Brain extracellular glucose increased after the transient elevation of plasma glucose following EN. These results suggest that brief, physiologic elevations in plasma glucose after EN produced no changes in brain extracellular glutamate concentration or lactate/pyruvate ratio. These data may help determine the plasma glucose levels most effective for avoiding brain metabolic acidosis in patients after SAH. It remains unclear, however, how SAH itself influences these findings. PMID- 19815234 TI - Fibrin sealant (Tisseel) for hiatal mesh fixation in an experimental model in pigs. AB - BACKGROUND: This study was designed to assess the efficacy of the fibrin sealant fixation of titanized polypropylene mesh in experimental hiatal mesh closure in pigs. Prosthetic hiatal closure is recommended for the repair of large hiatal/paraesophageal hernias as well as for antireflux surgery. However, only limited data exist on the favorable choice of meshes and fixation devices. Migration of the implant and trauma to neighboring organs due to perforating devices, such as sutures or tacks, present potentially lethal complications. In this study, we propose the fixation of titanized polypropylene meshes (TS) specifically developed for hiatal closure (TISure; GfE Medizintechnik GmbH, Nuremberg, Germany) with human fibrin sealant (FS, Tisseel; Baxter Biosciences, Vienna, Austria). MATERIALS AND METHODS: A laparotomy was carried out in 7 mini pigs (27-30 kg bodyweight) under general anaesthesia, and a TS was implanted after precise dissection of the right and left crura and the crural commissure. The key hole of the TS was placed around the esophagus at the gastroesophageal junction. One mL of FS was applied with the Easy Spray system (Baxter Biosciences, Vienna, Austria) for circular and three dimensional mesh fixation onto the diaphragm. Due to the lack of accepted gold standards of hiatal mesh reinforcement, no control group was used. Animals were sacrificed after 4 wk, and meshes were explanted after macroscopical assessment of the correct position and tissue integration. Histology was performed. RESULTS: All meshes showed excellent tissue integration and no signs of migration or dislocation. FS was completely degraded and replaced by well vascularized fibroblastic tissue. CONCLUSIONS: Titanized polypropylene mesh with FS fixation was found to be a safe and efficient combination for reinforcement of the hiatal closure in this preliminary experimental model. PMID- 19815235 TI - Motorcycle helmets save lives, but not limbs: a National Trauma Data Bank analysis of functional outcomes after motorcycle crash. AB - BACKGROUND: The incidence of motorcycle fatalities over the last 10 y has nearly doubled. There is considerable evidence that motorcycle helmets save lives, but there are little data regarding the impact that helmets have on functional outcomes after a motorcycle crash. The objective of this study was to determine the difference between helmeted and non-helmeted motorcyclists in the odds of developing a functional deficit at discharge in three domains: speech, locomotion, and feeding. METHODS: Reviewed cases in the National Trauma Databank v7.0 involved in motorcycle collisions. Multiple logistic regression was used to analyze the effect of helmets on mortality and functional outcomes, adjusting for age, race, gender, insurance status, anatomic and physiologic injury severity, and head injury. RESULTS: The adjusted odds of mortality (0.75; 95% CI 0.65-0.86) and functional deficits in speech (0.82; 95% CI 0.69-0.97), locomotion (1.19; 95% CI 1.11-1.29), and feeding (0.96 95% CI 0.84-1.08) among helmeted riders was compared with non-helmeted motorcyclists with equivalent injuries. CONCLUSION: Helmeted motorcyclists are less likely to die and develop a deficit in speech after a motorcycle collision. These data support that motorcycle helmets are important in preventing functional deficits related to head injury. PMID- 19815236 TI - Infarction induced myocardial apoptosis and ARC activation. AB - BACKGROUND: Apoptosis is thought to play a role in infarction induced ventricular remodeling. Apoptosis repressor with caspase recruitment domain (ARC) has been shown to limit cardiomyocytes apoptosis; however, its role in the pathogenesis of heart failure is not established. This study examines the regional and temporal relationships of apoptosis, ARC, and remodeling. METHODS: Myocardium was harvested from the infarct borderzone and remote regions of the left ventricle (LV) at 2 (n=8), 8 (n=6), and 32 (n=5) wk after MI. Activated ARC was compared with myocardial apoptosis in each region at each time. Both were then compared with the progression of remodeling. RESULTS: LV systolic volume increased by a factor 1.56+/-0.06 and 2.09+/-0.07 at 2 and 8 wk, respectively then stabilized by 32 wk (2.08+/-0.18). Activated ARC was elevated at 2 wk, diminished at 8 wk, and increased again at 32 wk in both regions. Apoptosis was elevated at 2 wk, and further increased at 8 wk. By 32 wk, apoptosis had diminished significantly. CONCLUSIONS: In a large animal infarction model, remodeling varied directly with the degree of apoptosis and inversely with ARC activation, suggesting that ARC acts as a natural regulatory phenomenon that limits apoptosis induced ventricular remodeling. PMID- 19815237 TI - American Ginseng inhibits induced COX-2 and NFKB activation in breast cancer cells. AB - BACKGROUND: Epidemiologic evidence suggests reduced breast cancer mortality in users of American Ginseng (AG) (Panax quinquefolium). We hypothesized that AG extract decreases proliferation of human breast cancer cells via an anti inflammatory effect applicable to the prevention of breast and other cancers. MATERIAL AND METHODS: A defined lyophilized aqueous extract of AG (LEAG) was dissolved in DMSO 1mg/mL, and serially diluted in saline. The cell lines MDA MB 231 and MCF7 were stimulated with the phorbol ester PDBu and treated with 100-500 mcg/mL LEAG. Proliferation was measured by MDA assay. Induced COX-2 expression was assayed by ELISA. Activation of NFkappaB by phosphorylation of the p65 subunit was quantified by CASE (cellular activation of signaling ELISA). RESULTS: Both cell lines had reduced proliferation when treated with LEAG. PDBu stimulation of MDA MB 231 increased expression of the COX-2 protein 20-fold at 48 hours (P<0.005). COX-2 protein expression remained at baseline concentrations in PDBu- treated MDA MB 231 cells exposed to 100 mcg/mL LEAG. The CASE assay showed a 4-fold increase in p65 activation 24 hours after PDBu treatment in normal medium, while phosphorylated p65 dropped below baseline in the cells treated with PDBu plus LEAG. CONCLUSION: In MDA MB 231, COX-2 was inducible with PDBu. This induced COX-2 expression was blocked by 100 microgram/mL LEAG in a time course consistent with the decline in the activated p65 subunit of NFkappaB. These results provide an anti-inflammatory mechanism for a possible anti-cancer effect of American Ginseng. PMID- 19815238 TI - Stomal complications in the newborn with necrotizing enterocolitis. AB - BACKGROUND: Infants who develop necrotizing enterocolitis (NEC) are usually managed with fecal diversion. The integrity of the bowel being diverted is often suboptimal. Our clinical impression is that stomas created in this circumstance are fraught with complications. The purpose of this study is to quantify the rate of these complications and identify risk factors. METHODS: A retrospective data collection from May 1999 to May 2008 on infants undergoing laparotomy for NEC was conducted. Data collected included gestational age, birth weight, age, and weight at operation, indications for surgical therapy, procedure performed, time to stoma output, time to takedown of stoma, complication directly related to the ostomy, and mortality. Data comparisons were analyzed statistically using chi(2), Pearson's correlation, Fisher's exact test, or a 2-tailed Student's t-test with significance reported for P<0.05. RESULTS: A total of 73 patients were identified. Mean gestational age was 28 (+/-4) wk, mean birth weight was 1247 (+/ 713) g. Mean age at the time of surgery was 23 (+/-27) d, and mean weight at operation was 1513 (+/-1306) g. The most common indication for surgical intervention was pneumoperitoneum (n=43, 58%). The most common level of intestinal diversion was the ileum (n=63, 85%). In-house mortality was 13%. There were 31 patients (42%) who developed 32 stoma-related complications. Demographic or preoperative variables that were a significant predictor of stoma-related complications were gestational age (P=0.003) and preoperative weight (P=0.024). CONCLUSION: Premature infants carry a risk for developing stoma-related complications. Within that cohort, there is significantly increased risk of stoma related complications in patients who are younger in gestational age and who have low preoperative weight. Future prospective studies may allow insight into preventative practices. PMID- 19815239 TI - Effective immune restoration after immunosuppressant discontinuation in a lupus patient presenting progressive multifocal leukoencephalopathy. AB - Progressive multifocal leukoencephalopathy (PML) is an opportunistic infection of the central nervous system with JC virus. Few cases have been described in lupus patients. We describe biopsy-proven PML in a lupus patient receiving mycophenolate mofetil and corticosteroids. Although the patient received no antiviral treatment, the polymerase chain reaction test for JC virus became negative in cerebrospinal fluid after immunosuppressant discontinuation and the patient survived. We discuss the restoration of immune efficiency after immunosuppressant discontinuation in this case and compare the clinical, radiological and histological features with the inflammatory PML form described in human immunodeficiency virus-infected patients. PMID- 19815241 TI - Acute health problems related to the operation mounted to clean the Hebei Spirit oil spill in Taean, Korea. AB - The authors investigated acute health problems in people engaged in the operation mounted to clear the Hebei Spirit oil spill which occurred in December 2007 in Taean County, South Korea, and identified the risk factors associated with the development of symptoms. Eight hundred forty-six people engaged in the clean up operation for periods between 7 and 14 days were examined. Demographic information and risk factors were obtained using a questionnaire. Symptoms were classified into six categories: back pain, skin lesions, headache, and eye, neurovestibular, and respiratory symptoms. Residents and volunteers engaged in the Hebei Spirit oil spill clean up operation experienced acute health problems. Risk analyses revealed that more frequent and greater exposure was strongly associated with a higher occurrence of symptoms. PMID- 19815242 TI - Effect of iron overload on glucose metabolism in patients with hereditary hemochromatosis. AB - Diabetes mellitus (DM) affects 30% to 60% of patients with hereditary hemochromatosis (HH). The underlying pathophysiology of DM in patients with hemochromatosis has not been fully elucidated. We studied both insulin secretion and insulin sensitivity in a cohort of patients with HH. We studied glucose metabolism in 53 newly diagnosed HH patients using a standard 75-g oral glucose tolerance test. Basal and stimulated insulin sensitivities were calculated using the quantitative insulin sensitivity check index and oral glucose insulin sensitivity index, respectively. beta-Cell function was assessed using C-peptide concentrations during the oral glucose tolerance test after adjusting for ambient insulin sensitivity. Twenty healthy subjects served as the control group. Fifteen subjects (28%) with HH had abnormal glucose tolerance (AGT). Seven (13%) had DM, and 8 (15%) had impaired glucose tolerance. As well as higher fasting glucose and glycated hemoglobin, those with AGT had a higher fasting insulin and C-peptide levels compared with those with normal glucose tolerance (NGT) (all Ps < .05). Insulin sensitivity measurements showed that the subjects in HH group with AGT were more insulin resistant than the subjects with NGT and controls subjects (P < .05). No significant changes were observed between the groups with NGT and AGT regarding hepatic insulin extraction and both indices related to insulin release in subjects with HH. Our cohort of patients with hemochromatosis and AGT had features similar to typical type 2 DM patients. These findings challenge the traditional view that DM in hemochromatosis is due primarily to iron-induced beta cell failure. PMID- 19815243 TI - Effects of rosiglitazone and metformin treatment on apelin, visfatin, and ghrelin levels in patients with type 2 diabetes mellitus. AB - Visfatin, ghrelin, and apelin are the most recently identified adipocytokines; but their response to insulin-sensitizing agents is poorly clarified. We aimed to assess the differential effects of either rosiglitazone or metformin monotherapy on the aforementioned adipocytokines in patients with type 2 diabetes mellitus (T2DM). One hundred T2DM patients (30 men, 70 women), with poor glycemic control (glycosylated hemoglobin >6.5%) while taking 850 mg of metformin daily, were enrolled. All participants were randomized to receive either adjunctive therapy with rosiglitazone (8 mg/d, n = 50) or the maximum dose (2550 mg/d) of metformin (MET group, n = 50). Anthropometric parameters, glycemic and lipid profile, high sensitivity CRP (hs-CRP), insulin resistance (homeostasis model assessment of insulin resistance index [HOMA-IR]), visfatin, ghrelin, and apelin were assessed at baseline and after 14 weeks of therapy. Both rosiglitazone and metformin led to similar, significant improvement in glycemic profile and apelin levels, whereas lipid parameters, fat mass, and visfatin remained almost unaffected (P > .05). Insulin resistance was significantly attenuated in both groups, but to a lesser degree in the MET group (P = .045). Rosiglitazone-treated patients experienced a significant decrease in hs-CRP and systolic blood pressure compared with baseline values and those of the MET group (P < .05). Besides, rosiglitazone treatment considerably increased plasma ghrelin (3.74 +/- 1.52 ng/mL) in comparison with either baseline (P = .034) or metformin monotherapy values (-2.23 +/- 1.87 ng/mL, P = .008). On the other hand, the MET group, rather than the rosiglitazone group, had decreased body mass index (-0.79 +/- 0.47 vs 0.56 kg/m(2), P = .009). The aforementioned changes in apelin and ghrelin were independently associated with HOMA-IR changes. Both rosiglitazone and metformin favorably changed glycemic indexes and apelin levels. The addition of rosiglitazone seemed to confer greater benefits in ghrelin, hs-CRP, systolic blood pressure, and HOMA-IR regulation than metformin monotherapy. Although these results reflect improvement in cardiovascular risk profile, the overall clinical importance of insulin sensitizers must be further assessed. PMID- 19815240 TI - Cardiovascular event rates in patients with cerebrovascular disease and atherothrombosis at other vascular locations: results from 1-year outcomes in the Japanese REACH Registry. AB - The REduction of Atherothrombosis for Continued Health (REACH) Registry is a large, international, prospective cohort of patients with atherothrombosis or multiple (>or=3) risk factors (MRFs) for atherothrombosis. Japanese patients (n=5193) were enrolled into the REACH registry between August and December 2004. One-year event rate in patients with cerebrovascular disease (CVD) was compared with that of patients with symptomatic atherothrombosis at other locations. After one year (n=5021), patients with CVD (n=1962) experienced a higher rate of non fatal strokes than patients with coronary artery disease (CAD), peripheral artery disease (PAD) or MRFs alone (2.77% vs. 1.28%, 2.07% and 1.56%, respectively), but a lower rate of non-fatal myocardial infarction (0.45% vs. 1.31%, 0.77% and 0.66%, respectively). Patients with CVD plus disease in >or=1 other vascular bed had higher rates of cardiovascular events than patients with CVD alone. Overall, event rates including non-fatal stroke, non-fatal myocardial infarction and cardiovascular death were higher for patients with CVD and PAD than for patients with CVD and CAD. Asymptomatic carotid stenosis >or=70% and ankle-brachial index <0.9 were significant predisposing factors for stroke. Patients with CVD and co existing atherothrombotic diseases had a high risk of recurrent events, including events arising in other vascular beds than originally diagnosed. PMID- 19815244 TI - Tiglicamides A-C, cyclodepsipeptides from the marine cyanobacterium Lyngbya confervoides. AB - The Floridian marine cyanobacterium Lyngbya confervoides afforded cyclodepsipeptides, termed tiglicamides A-C (1-3), along with their previously reported analogues largamides A-C (4-6), all of which possess an unusual tiglic acid moiety. Their structures were deduced by one- and two-dimensional NMR combined with mass spectrometry and the absolute configurations established by chiral HPLC and Marfey's analysis of the degradation products. Compounds 1-3 moderately inhibited porcine pancreatic elastase in vitro with IC(50) values from 2.14 to 7.28 microM. Compounds 1-6 differ from each other by one amino acid residue within the cyclic core structure, suggesting an unusually relaxed substrate specificity of the nonribosomal peptide synthetase that is the putative biosynthetic enzyme responsible for the corresponding amino acid incorporation. PMID- 19815245 TI - Hydroxylation of the diterpenes ent-kaur-16-en-19-oic and ent-beyer-15-en-19-oic acids by the fungus Aspergillus niger. AB - The diterpenes ent-kaur-16-en-19-oic acid (1) and ent-beyer-15-en-19-oic acid (2) are the major constituents of a spasmolytic diterpenic mixture obtained from the roots of Viguiera hypargyrea, a Mexican medicinal plant. Microbial transformation of 1 and 2 was performed with Aspergillus niger. Two metabolites, ent 7alpha,11beta-dihydroxy-kaur-16-en-19-oic acid (4) and ent-1beta,7alpha-dihydroxy kaur-16-en-19-oic acid (5), were isolated from the incubation of 1, and one metabolite, ent-1beta,7alpha-dihydroxy-beyer-15-en-19-oic acid (6), was isolated in high yield (40%) from 2. The structures were elucidated on the basis of spectroscopic analyses and confirmed by X-ray crystallographic studies. Compounds 1-4 and 6 and methyl ester derivatives 4a and 6a were evaluated for their ability to inhibit the electrically induced contraction of guinea-pig ileum. Compounds 1, 3, 4, 4a and 5 were significantly active. These results showed that dihydroxylation of 1 at 7beta, 11alpha-, and 1alpha, 7beta-positions resulted in a loss of potency. PMID- 19815246 TI - [Deep brain stimulation and gait disorders in Parkinson disease]. AB - INTRODUCTION: Gait disorders and freezing of gait (FOG) are seen in most patients with advanced Parkinson disease. Response to levodopa and deep brain stimulation is variable across patients. STATE OF ART: Thalamic stimulation is ineffective on gait and can even worsen balance when bilaterally applied. Pallidal stimulation moderately improves gait disorders and FOG although this effect tends to wane after three to five years. Stimulation of the subthalamic nucleus (STN) improves levodopa-responsive gait disorders and FOG. However, some patients worsen after STN stimulation and others are better improved under levodopa than under STN stimulation. Synergistic effects of the two treatments have been reported. As for pallidal stimulation, there is a failure of long-term STN stimulation to improve gait, probably due to the involvement of non-dopaminergic pathways as the disease progresses. Levodopa-resistant gait disorders and FOG do not usually benefit from STN stimulation. In the rare cases of levodopa-induced FOG, STN stimulation may be indirectly effective, as it enables reduction or arrest of the levodopa treatment. PERSPECTIVES: Pedunculopontine nucleus stimulation has recently been performed in small groups of patients with disabling gait disorders and FOG. Although encouraging, the first results need to be confirmed by controlled studies involving larger series of patients. CONCLUSIONS: Overall, gait disorders remain a motor PD symptom that is little improved, or only temporarily, by current pharmacological and surgical treatments. Patient management is complex. PMID- 19815247 TI - Effect of trimerization motifs on quaternary structure, antigenicity, and immunogenicity of a noncleavable HIV-1 gp140 envelope glycoprotein. AB - The external domains of the HIV-1 envelope glycoprotein (gp120 and the gp41 ectodomain, collectively known as gp140) contain all known viral neutralization epitopes. Various strategies have been used to create soluble trimers of the envelope to mimic the structure of the native viral protein, including mutation of the gp120-gp41 cleavage site, introduction of disulfide bonds, and fusion to heterologous trimerization motifs. We compared the effects on quaternary structure, antigenicity, and immunogenicity of three such motifs: T4 fibritin, a GCN4 variant, and the Escherichia coli aspartate transcarbamoylase catalytic subunit. Fusion of each motif to the C-terminus of a noncleavable JRCSF gp140(-) envelope protein led to enhanced trimerization but had limited effects on the antigenic profile and CD4-binding ability of the trimers. Immunization of rabbits provided no evidence that the trimerized gp140(-) constructs induced significantly improved neutralizing antibodies to several HIV-1 pseudoviruses, compared to gp140 lacking a trimerization motif. However, modest differences in both binding specificity and neutralizing antibody responses were observed among the various immunogens. PMID- 19815248 TI - Impact of microfiltration treatment of secondary wastewater effluent on biofouling of reverse osmosis membranes. AB - The effects of microfiltration (MF) as pretreatment for reverse osmosis (RO) on biofouling of RO membranes were analyzed with secondary wastewater effluents. MF pretreatment reduced permeate flux decline two- to three-fold, while increasing salt rejection. Additionally, the oxygen uptake rate (OUR) in the biofouling layer of the RO membrane was higher for an RO system that received pretreated secondary wastewater effluent compared to a control RO system that received untreated secondary effluent, likely due to the removal of inert particulate/colloidal matter during MF. A higher cell viability in the RO biofilm was observed close to the membrane surface irrespective of pretreatment, which is consistent with the biofilm-enhanced concentration polarization effect. Bacterial 16S rRNA gene clone library analysis revealed dominant biofilm communities of Proteobacteria and Bacteroidetes under all conditions. The Cramer-von Mises test statistic showed that MF pretreatment did not significantly change the bacterial community structure of RO membrane biofilms, though it affected bacterial community structure of non-membrane-associated biofilms (collected from the feed tank wall). The finding that the biofilm community developed on the RO membrane was not influenced by MF pretreatment may imply that RO membranes select for a conserved biofilm community. PMID- 19815249 TI - Impacts of salinity on the performance of high retention membrane bioreactors for water reclamation: A review. AB - Recent efforts in the field of used water treatment and water reclamation have led to the development of a number of innovative high retention membrane bioreactor (HRMBR) systems. These systems invariably combine a high rejection membrane separation with a biological treatment. A common positive outcome of these systems is that smaller size organic contaminants are effectively retained, which facilitates their biodegradation and thus produces high quality product water. This provides the desired high level of separation, but also leads to salt accumulation with potentially adverse effects on the operations. The effects of elevated salt condition are complex, and impact on aspects covering physicochemical parameters, microbiology and membrane performance. The salt concentration factor is an important operating parameter to be optimised in the HRMBR systems. This paper aims to elucidate the important issues associated with the use of HRMBR systems under elevated salt conditions up to 50gL(-1). PMID- 19815250 TI - Cost and effluent quality controllers design based on the relative gain array for a nutrient removal WWTP. AB - The main objective of this work was the design of different effluent quality controllers and a cost controller for WWTPs. This study was based on the relative gain array (RGA) analysis applied to an anaerobic/anoxic/aerobic (A(2)/O) configuration of a simulated WWTP, with combined removal of organic matter, nitrogen and phosphorus. The RGA analysis was able to point out the best pairing amongst the input and the output control variables of the plant to design low order and decentralized effluent quality controllers, such as proportional integral controllers for each variable of interest (ammonium, nitrate and phosphate). In a second step, a cost controller to automatically search for the most economic setpoints of the effluent quality controllers was implemented based on the best decentralized control structure tested. The simulated plant was operated under different control modes that chronologically represent control configurations becoming gradually more complex: (i) in open loop; (ii) with dissolved oxygen (DO) control in the last aerobic reactor only; (iii) with the effluent quality controllers active; (iv) with the effluent quality controllers active and automatically receiving the setpoints from a cost controller. The effluent quality controllers alone and the cost control together with effluent quality controllers could save up to 42,000 Euros/year and 225,000 Euros/year, respectively, when compared to the operating costs of the plant operating with DO control (a reduction of 2.5% and 13% of the operating costs, respectively). The cost controller proved to be a good tool for automating the search of the most profitable setpoints of the effluent quality controllers for a given cost setpoint. PMID- 19815251 TI - Psychiatric pharmaceuticals in the environment. AB - Psychiatric pharmaceuticals, such as anxiolytics, sedatives, hypnotics, and antidepressants, are among the most prescribed active substances throughout the world. The occurrence of these widely used compounds in environmental matrices (wastewaters, surface, ground and drinking waters, soils, sediments, bio-solids and tissue), as well as the first studies indicating their high persistence and toxicity to non-target organisms, justify the growing concern about these emerging environmental pollutants. Despite this increasing interest, there is a considerable lack of knowledge about the environmental fate of a large number of psychiatric pharmaceuticals and further research about this topic is needed. This paper aims to review the literature data related to the occurrence, persistence, environmental fate and toxicity for non-target organisms of this group of pharmaceuticals. The analytical methods developed for the determination of psychiatric medicines in environmental matrices are also highlighted. PMID- 19815252 TI - PCDD/Fs in ambient air in north-east Italy: the role of a MSWI inside an industrial area. AB - The stack gases of a municipal solid waste incinerator (MSWI), and ambient air were sampled in four locations around the plant for the analysis of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDDs/Fs). The sampling area was close to an industrial area near Trieste, in north-east Italy. The purpose of the study was to estimate the impact of the MSWI emissions and to distinguish the contribution of these emissions from other potential emission sources in the industrial area. PCDD/F atmospheric concentrations were similar to those generally detected in urban-rural areas with one location about 2-3 times more contaminated than the others. Since the most contaminated location was inside the industrial area but upwind of the MSWI, principal component analysis (PCA) was used to establish whether other sources were the cause. This analysis clearly showed that a local steel plant's emission was the main source of PCDDs/Fs in ambient air. This study highlights the usefulness of multivariate data analysis such as PCA to identify, among different potential emission sources, the one really responsible for the contamination. PMID- 19815253 TI - Brominated flame retardants in the Arctic environment--trends and new candidates. AB - Polybrominated diphenyl ethers (PBDEs) containing two to 10 bromines are ubiquitous in the Arctic, in both abiotic and biotic samples. Hexabromocyclododecane (HBCD) is also ubiquitous in the Arctic, with the gamma HBCD isomer predominating in air, the alpha-HBCD isomer predominating in biota and similar concentrations of alpha-, beta- and gamma-HBCD found in marine sediments. Other brominated flame retardants (BFRs) found in some Arctic samples are polybrominated biphenyls (PBBs), tetrabromobisphenol A (TBBPA), 1,2-bis(2,4,6 tribromophenoxy)ethane (BTBPE), hexabromobenzene (HxBBz), pentabromoethylbenzene (PBEB), pentabromotoluene (PBT), and 1,2-dibromo-4-(1,2-dibromoethyl)cyclohexane (TBECH). Temporal trends of tetra- to heptaBDEs and HBCD show increasing concentrations or a tendency to levelling off depending on the matrix (air, sediment, biota) and location, but no uniform picture for the Arctic emerges. BDE 209 concentrations are increasing in air. PBDEs and HBCD spatial trends in seabirds and marine mammals are similar to those seen previously for polychlorinated biphenyls (PCBs), with highest concentrations found in organisms from East Greenland and Svalbard. These trends indicate western Europe and eastern North America as important source regions of these compounds via long range atmospheric transport and ocean currents. Latitudinal trends showed lower concentrations and fluxes of PBDEs at higher latitudes. The tetra-hexaBDEs and alpha-HBCD biomagnify in Arctic food webs. Results for BDE-209 are more conflicting, showing either only low or no biomagnification potential. PBDE and HBCD concentrations are lower in terrestrial organisms and higher in marine top predators such as some killer whale populations in Alaska and glaucous gulls from the Barents Sea area. Higher concentrations are seen near populated areas indicating local sources. Findings of BTBPE, HxBBz, PBEB, PBT and TBECH in seabirds and/or marine mammals indicate that these compounds reach the Arctic, most probably by long range atmospheric transport and accumulate in higher trophic level organisms and that increasing use as PBDE replacements will lead to increasing concentrations. PMID- 19815254 TI - Atmospheric levels of BTEX compounds during the 2008 Olympic Games in the urban area of Beijing. AB - The hourly concentrations of BTEX (Benzene, Toluene, Ethylbenzene, m,p-Xylene and o-Xylene) in the urban area of Beijing were measured during July-October 2008, covering the periods of the 2008 Olympic Games and Paralympic Games. The atmospheric BTEX were pre-concentrated on Tenax-TA tubes, and analyzed by GC-PID (Gas Chromatography with Photo Ionization Detector) after thermal desorption. During the games, the mean daytime concentrations of benzene, toluene, ethylbenzene, m,p-xylene and o-xylene were 2.37, 3.97, 1.92, 3.51 and 1.90 microg/m3, respectively, and were 52.8%, 63.9%, 56.4%, 56.8% and 46.9%, respectively lower than those after the games. The significantly positive correlation between BTEX and CO as well as the ratio of benzene/toluene suggested that the vehicle exhaust was the major source of BTEX during the whole investigated period. The extremely high ratios of ethylbenzene to m,p-xylene (E/X) were mainly observed at noontime in haze days, indicating that photochemical reactions were highly active under these typical days. PMID- 19815255 TI - YouTube as source of prostate cancer information. AB - OBJECTIVES: Patients can search the Internet for prostate cancer information, and YouTube is a popular Web site that they may consult. We analyzed the prostate cancer videos on YouTube for information content and the presence of bias. METHODS: YouTube was searched for videos about prostate-specific antigen (PSA) testing, radiotherapy, and surgery for prostate cancer. The included videos were in English and <10 minutes long. Two physician viewers watched each video and assigned a score for information content (excellent, fair, poor) and bias (for, against, neutral, or balanced). A third viewer arbitrated any discrepancies. The kappa statistic was used to measure interobserver variability, and Pearson's test was used to assess correlation. RESULTS: A total of 14 PSA videos, 5 radiotherapy videos, and 32 surgery videos were analyzed. The PSA testing videos averaged 1480 +/- 2196 views and 146 +/- 174 s long and had an average viewer rating of 3.1 +/- 2.1 (viewer rating scale 0-5). The surgery videos averaged 2044 +/- 3740 views and 172 +/- 122 s long and had an average viewer rating of scored 3 +/- 2.2. The radiotherapy videos averaged 287 +/- 255 views and 97 +/- 45 s long and had a score of 1.8 +/- 2.5. The information content was fair or poor for 73% of all videos. The bias for surgery, radiotherapy, or PSA testing was present in 69% of videos; 0% of videos were biased against treatment or PSA testing. The interobserver variability was well above than expected by chance alone. CONCLUSIONS: The results of our study have shown that although some videos are robust sources of information, given the preponderance of modest and unbalanced information among reviewed videos, YouTube is an inadequate source of prostate cancer information for patients. PMID- 19815256 TI - The effect of testosterone replacement therapy on bladder functions and histology in orchiectomized mature male rats. AB - OBJECTIVE: To investigate the effect of testosterone replacement therapy on bladder functions and smooth muscle/collagen content in orchidectomized orchiectomized mature male rats. METHODS: The study included 25 mature male Sprague-Dawley rats divided into 3 groups. After bilateral orchiectomy, 8 rats received intramuscular saline injection, as a control group, and 8 rats received intramuscular injection of testosterone undecanoate 100 mg/kg as a treatment group. The sham group had 9 rats. Urodynamic studies were performed in all groups, before and after the study. The rats were killed after 60 days, and cystometric findings and smooth muscle/collagen ratio of the bladders were compared between the groups. RESULTS: From the beginning to the end of the experiment, mean maximal bladder capacity increased 46.61% +/- 20.82 in the testosterone treatment group, while decreased 38.91% +/- 17.83 in control group, revealing a significant difference (P = .002). Smooth muscle/collagen ratio was significantly higher in the testosterone treatment group (1.53 +/- .34) than in the control group (1.05 +/- .32), (P = .01). CONCLUSIONS: This study showed that bladder capacity and smooth muscle/collagen content improved with testosterone therapy in orchiectomized rats. Therefore, testosterone replacement therapy in late-onset hypogonadal men with urogenital dysfunction may have a positive role to improve bladder function by increasing bladder smooth muscle. PMID- 19815257 TI - Single-port transvesical excision of foreign body in the bladder. AB - OBJECTIVES: To present a novel technique to remove intravesical polypropylene mesh through a single laparoscopic port placed directly into the bladder. METHODS: A Triport single-site access system was placed transvesically and carbon dioxide was used for insufflation of the bladder. A combination of straight and articulating laparoscopic instruments was used to dissect the mesh away from the bladder mucosa and transect each end for complete removal of foreign bodies. Mucosal reapproximation was performed on the latter case. RESULTS: Two patients were managed adequately in the outpatient setting. No suprapubic catheters were necessary, and patients were discharged within 23 hours. CONCLUSIONS: Removal of foreign bodies of the bladder through a single transvesical laparoscopic port is technically feasible. This procedure offers excellent visualization of mesh material, especially near the bladder neck where these foreign bodies often reside. This approach offers patients a minimally invasive approach through a single small incision. PMID- 19815258 TI - Bacterial sepsis after prostate biopsy--a new perspective. AB - OBJECTIVES: To determine the incidence of sepsis following transrectal ultrasound (TRUS)-guided prostate biopsy at our center. METHODS: We retrospectively reviewed a group of 24 men who presented with urosepsis after undergoing TRUS biopsy at our center. RESULTS: Of the 24 men, 22 were given prophylactic ciprofloxacin. The median time to presentation of sepsis was 1 day after biopsy. The median length of hospitalization was 4 days. Escherichia coli was the most frequent cause of urosepsis (67%). Variable resistance patterns were observed. Enterobacter cloacae and Streptococcus viridans were isolated in 2 cases. No bacteria were isolated in 6 cases. Two patients who received extensive antibiotic prophylaxis still developed urosepsis. Treatment of patients infected with multiresistant anaerobic strains using metronidazole among others, proved successful. High sensitivities toward cefazolin, gentamicin, and tobramycin were observed. The number of cases reported was likely an underestimation, because some patients may have reported to other hospitals and were not captured by this study. In addition, some patients may not have developed infection and urosepsis despite harboring ciprofloxacin-resistant bacteria. CONCLUSIONS: Prophylactic ciprofloxacin is still a useful option for the prevention of urosepsis after TRUS biopsy, as the incidence is relatively low. For the patient who develops urosepsis after TRUS biopsy, ciprofloxacin resistance needs to be suspected and the treatment regime should be tailored to the resistance profiles of the local region, the patient's medical history, and the culture and sensitivity reports. PMID- 19815259 TI - Public survey and survival data do not support recommendations to discontinue prostate-specific antigen screening in men at age 75. AB - OBJECTIVES: To evaluate the US Preventative Services Task Force (USPSTF) recommendation to discontinue prostate-specific antigen (PSA) screening at age 75. METHODS: Public survey: A cohort of 340 patients was surveyed at our PSA screening clinic and stratified by awareness of the recommendation and education level. Age (< 75, >or= 75), race, health insurance status, knowledge of prostate cancer, and opinion on screening discontinuation at age 75 was evaluated between groups. Disease risk and survival analysis: A cohort of 4196 men who underwent radical prostatectomy between 1988 and 2008 was stratified into age groups: < 65, 65-74, and >or= 75. Associations between clinicopathologic variables, disease risk, and survival were compared between age groups using univariate and multivariate analysis. RESULTS: Approximately 78% of men surveyed disagreed with the USPSTF recommendation. The number of men who disagreed was not significantly different between awareness groups (P = .962). Awareness of new screening guidelines showed a significant difference (P = .006) between education groups. Age >or= 75 years was predictive of high-risk disease based on D'Amico's criteria (odds ratio = 2.72, P = .003). Kaplan-Meier and Cox regression analyses showed an association of men aged >or= 75 years with higher rate of PSA recurrence, distant metastasis, and disease specific death compared with the age groups of < 65 and 65-74 (P <.05). CONCLUSIONS: Men presenting to our PSA screening clinic disagreed with discontinuation of screening at age 75. Men aged >or= 75 years had higher risk disease and poorer survival. The USPSTF recommendation was supported neither by public opinion nor disease risk and survival results. PMID- 19815260 TI - Percutaneous renal cryoablation of angiomyolipomas in patients with solitary kidneys. AB - OBJECTIVES: To determine the feasibility and safety of performing percutaneous cryoablation of angiomyolipomas (AMLs) in patients with solitary kidneys. METHODS: Three patients with AMLs involving a solitary kidney underwent computed tomography-guided percutaneous cryoablation. All lesions were located in the lateral/posterior part of the kidney, allowing for safe access from the skin for cryoprobe insertion. Intravenous sedation and local anesthesia were used for each patient. Follow-up computed tomography or magnetic resonance imaging and physical examination, urinalysis, and serum blood urea nitrogen/creatinine measurement were performed to evaluate for lesion recurrence and to evaluate the safety profile. RESULTS: Three tumors (1.2-2.5 cm) were treated. The patients experienced minimal to no pain during percutaneous cryoablation, and all were discharged the same day. No procedural or postoperative complications were noted. During the follow-up period (5-36 months), the first 2 patients had no radiographic evidence of recurrence. Initial follow-up imaging of the third patient displayed persistent AMLs. CONCLUSIONS: A review of the published data suggested the necessity to prophylactically treat AMLs of solitary kidneys. In this series, percutaneous cryoablation proved a safe and effective method for treating these lesions. This ultimately provides a minimally invasive option for similar patients, potentially avoiding an open surgical procedure or the risk of hemorrhage. PMID- 19815261 TI - Retroperitoneal laparoscopic live donor nephrectomy: a cost-effective approach. AB - OBJECTIVES: To establish a modified cost-effective and safe approach of retroperitoneal laparoscopic live donor nephrectomy (RPLLDN). METHODS: Between December 2003 and December 2008, total of 109 living related renal donors underwent consecutive three-port RPLLDNs. The initial retroperitoneal space was created by insertion of a rubber catheter attached to a saline-filled midfinger of a glove. The renal hilum and ureter were circumferentially mobilized. The tributaries of renal vessels were divided by harmonic scalpel without any clips. In laparoscopy, the ureter was sheared with scissors. A longitudinal 6-8 cm skin incision was enlarged form the primary trocar distally to retrieve the graft. The main renal vessels were controlled, using 2 Hem-o-lok clips placed at each proximal ends. The graft was retrieved manually through the skin incision. RESULTS: All the 109 RPLLDNs were carried out successfully. The mean operation time and mean warm ischemic time was 129.7 +/- 42.6 and 3.6 +/- 1.2 minutes, respectively. The mean blood loss was 73.6 +/- 53.7 mL. No blood transfusion or open conversion was required. No major complication occurred in the donors, and only 5% of the donors suffered from minor complications. A total of 3% and 4% of the recipients developed major and minor complications, respectively, and 12.8% of the donors required analgesics. The mean level of postoperative serum creatinine of the donors was 1.31 +/- 0.22 mg/dL and the mean level of postoperative serum creatinine of the recipients at the first month was 1.59 +/- 0.91 mg/dL. CONCLUSIONS: The modified approach of RPLLDN could be a cost effective and safe alternative for developing countries. PMID- 19815262 TI - Surgical treatment of renal cell carcinoma in the immunocompromised transplant patient. AB - OBJECTIVES: To determine whether iatrogenic immunosuppression used after transplantation infers a poor prognosis of renal cell carcinoma (RCC) as natural negative immune regulators have been associated with decreased cancer-specific survival from RCC. METHODS: All patients with a solid organ transplant who underwent radical nephrectomy or nephron-sparing surgery for nonhereditary sporadic RCC from 1970 to 2003 were identified and retrospectively reviewed. RESULTS: We identified 17 patients with surgically treated rcc who also underwent a solid organ transplant: 11 with transplant before RCC and 6 with transplant after RCC. Type of transplant included 9 kidney, 3 heart, 3 liver, 1 kidney and liver, and 1 kidney and pancreas. Tumor pathology included 10 clear-cell RCC and 7 papillary RCC. At the last follow-up 6 patients died at a mean of 5.9 years after nephrectomy. Among the 11 patients still alive, mean follow-up was 7.6 years. Only 1 patient died of RCC. This patient had metastatic clear-cell RCC that was completely resected 8 years before renal transplant. He had a recurrence 2 years post transplant and died 3 years after recurrence. No other patients experienced local or distant disease recurrence. Immunosuppression was decreased in only 2 patients; one secondary to RCC metastases and another for recurrent skin cancer. CONCLUSIONS: Surgical resection of RCC in transplant patients is associated with a low rate of progression, despite optimal immunosuppression. We recommend surgical resection of low-risk, organ-confined RCC without reduction in immunosuppression in patients with solid organ transplants. PMID- 19815263 TI - Dornier Lithotripter S 220 F EMSE: the first report of over 1000 treatments. AB - OBJECTIVE: To assess the short-term efficacy and safety of extracorporeal shock wave lithotripsy (ESWL) with the standard Dornier Lithotripter S 220 F EMSE in the treatment of a large population of 736 patients with renal and ureteral stones. This is the only report with more than 1000 treatments performed with this device. METHODS: Between January 2003 and July 2006, a total of 479 renal and 257 ureteral stones were treated with 1168 ESWL sessions. ESWL was performed on an outpatient basis. Patients were evaluated after 1 and 3 months. Stone size and location, total number of shockwaves, stone-free rate, and complications were investigated. RESULTS: The stone-free rate for renal calculi was 60.5% at 1 month and 82.5% at 3 months. The stone-free rate for ureteral stones was 58% at 1 month and 82.9% at 3 months. The overall stone-free rate was 59.6% at 1 month and 82.5% at 3 months. Anesthesia was not needed in any case. Analgesia with hospital admission was necessary in 15 patients (2.0%). The major complications observed were renal hematoma in only 1 patient (0.1%), obstruction with sepsis in 3 patients (0.4%), and steinstrasse development in 5 patients (0.7%). CONCLUSIONS: The Dornier Lithotripter S 220 F EMSE is a safe and effective tool in the treatment of urolithiasis; the lack of invasiveness and absence of anesthesia confirm its worthy role as an alternative to ureterorenoscopy and percutaneous nephrolitotomy. PMID- 19815264 TI - A prospective randomized study comparing shock wave lithotripsy and semirigid ureteroscopy for the management of proximal ureteral calculi. AB - OBJECTIVES: To conduct a prospective randomized study comparing both techniques for the management of solitary radio-opaque upper ureteral stones < 2 cm in diameter. The ideal treatment for upper ureteral stones > 1 cm size remains to be determined with shock wave lithotripsy (SWL) and ureteroscopy (URS) being acceptable options. METHODS: A total of 200 patients were included in the study. They were randomized into 2 equal groups. Group A underwent in situ SWL as a primary therapy. Group B underwent URS, using semirigid URS with intracorporeal lithotripsy. Efficiency quotient (EQ), cost analysis, and predictors of failure were estimated for both techniques. RESULTS: For stones of size > or = 1 cm, the initial stone-free rate for URS and SWL was 88% and 60%, respectively. The estimated EQ was 0.79 and 0.43 for both techniques respectively. For stones < 1 cm, the initial stone-free rate for URS and SWL was 100% and 80%, respectively. The estimated EQ was 0.88 and 0.70 for both techniques, respectively. The mean cumulative costs were significantly more in SWL group (P <.05). Predictors of URS failure included; male gender, failure to pass guidewire beyond the stone, and extravasation. Predictors of SWL failure included large stone size > 1 cm, calcium oxalate monohydrate stone, and higher degrees of hydronephrosis. CONCLUSIONS: URS with intracorporeal lithotripsy is an acceptable treatment modality for all proximal ureteral calculi, particularly stones > 1 cm. SWL should remain the first-line therapy for proximal ureteral calculi < or = 1 cm because of the less invasive nature and lower anesthesia (i.v. sedation). PMID- 19815265 TI - Silodosin in the treatment of the signs and symptoms of benign prostatic hyperplasia: a 9-month, open-label extension study. AB - OBJECTIVES: To evaluate long-term safety of the highly selective alpha(1A) adrenoceptor antagonist silodosin in men with signs and symptoms of benign prostatic hyperplasia (BPH). METHODS: Patients enrolled in this open-label extension study had completed 1 of 2 identical double-blind, placebo-controlled 12-week studies of silodosin treatment for symptomatic BPH. For 40 weeks, patients received silodosin 8 mg once daily with breakfast. Adverse events (AEs) were recorded to evaluate safety. Change in International Prostate Symptom Score was a secondary variable. RESULTS: Of the 661 participants, 435 (65.8%) completed the study; 431 patients (65.2%) experienced 924 AEs. No serious AEs that were considered drug-related by investigators occurred. AEs reported most often (percentage of patients) were retrograde ejaculation (20.9%), diarrhea (4.1%), and nasopharyngitis (3.6%). Orthostatic hypotension and dizziness occurred in 2.6% and 2.9% of patients, respectively. The percentage of patients with treatment-emergent AEs, stratified by preceding double-blind treatment (placebo or silodosin), was higher for de novo (previous treatment with placebo, 71.5%) than for continuing silodosin treatment (58.3%). More patients receiving de novo (7.5%) vs continuing treatment (1.9%) discontinued study participation because of retrograde ejaculation. Mean International Prostate Symptom Score change (standard deviation) from baseline to week 40 (observed cases) was -4.5 (6.7) for de novo treatment (P <.0001) and -1.6 (6.0) for continuing treatment (P <.01). CONCLUSIONS: Silodosin was well tolerated by men with BPH-related symptoms and was associated with low incidences of dizziness and orthostatic hypotension. Discontinuation because of retrograde ejaculation was more common among patients receiving silodosin de novo than in those who continued silodosin treatment. PMID- 19815266 TI - Testicular development and establishment of spermatogenesis in Nili-Ravi buffalo bulls. AB - Fifteen longitudinally reared Nili-Ravi buffalo bulls (Bubalus bubalis) were slaughtered at 1, 6, 12, 18, and 24 mo of age (n=3 per group) to observe testicular development and to examine qualitatively the establishment of spermatogenesis. With the age held constant, scrotal circumference and testes weight were correlated (0.95; P<0.05). Testes weight increased from 3.5+/-0.7 at 1 mo of age to 185+/-30g at 24 mo of age. Seminiferous tubules diameter developed in a linear fashion (57microm at 1 mo and 178microm at 24 mo), and the lumen formed at 12 mo of age. Differentiation of basal indifferent supporting cells to Sertoli cells started at 6 mo, and formation of Sertoli cells completed near 12 mo of age. Gonocytes predominated at 1 mo, but by 12 mo, most had been replaced by spermatogonia, thus rapid proliferation of tubular contents occurred at 12 mo (testes weight=75g). Spermatocytes were first observed at 12 mo, and their number increased through 18 and 24 mo. Establishment of spermatogenesis, as reflected by appearance of significant number of spermatids, occurred by 18 mo of age (testes weight 122g). Thus, the establishment of spermatogenesis was progressive from birth, and marked changes were observed during the last 6 mo. PMID- 19815267 TI - A lexical basis for N400 context effects: evidence from MEG. AB - The electrophysiological response to words during the 'N400' time window (approximately 300-500 ms post-onset) is affected by the context in which the word is presented, but whether this effect reflects the impact of context on access of the stored lexical information itself or, alternatively, post-access integration processes is still an open question with substantive theoretical consequences. One challenge for integration accounts is that contexts that seem to require different levels of integration for incoming words (i.e., sentence frames vs. prime words) have similar effects on the N400 component measured in ERP. In this study we compare the effects of these different context types directly, in a within-subject design using MEG, which provides a better opportunity for identifying topographical differences between electrophysiological components, due to the minimal spatial distortion of the MEG signal. We find a qualitatively similar contextual effect for both sentence frame and prime-word contexts, although the effect is smaller in magnitude for shorter word prime contexts. Additionally, we observe no difference in response amplitude between sentence endings that are explicitly incongruent and target words that are simply part of an unrelated pair. These results suggest that the N400 effect does not reflect semantic integration difficulty. Rather, the data are consistent with an account in which N400 reduction reflects facilitated access of lexical information. PMID- 19815269 TI - Treatment of clinically isolated syndrome: to be PreCISe. PMID- 19815268 TI - Effect of glatiramer acetate on conversion to clinically definite multiple sclerosis in patients with clinically isolated syndrome (PreCISe study): a randomised, double-blind, placebo-controlled trial. AB - BACKGROUND: Glatiramer acetate, approved for the treatment of relapsing-remitting multiple sclerosis, reduces relapses and disease activity and burden monitored by MRI. We assessed the efficacy of early treatment with glatiramer acetate in delaying onset of clinically definite multiple sclerosis. METHODS: In this randomised, double-blind trial, undertaken in 80 sites in 16 countries, 481 patients presenting with a clinically isolated syndrome with unifocal manifestation, and two or more T2-weighted brain lesions measuring 6 mm or more, were randomly assigned to receive either subcutaneous glatiramer acetate 20 mg per day (n=243) or placebo (n=238) for up to 36 months, unless they converted to clinically definite multiple sclerosis. The randomisation scheme used SAS-based blocks stratified by centre, and patients and all personnel were masked to treatment assignment. The primary endpoint was time to clinically definite multiple sclerosis, based on a second clinical attack. Analysis was by intention to treat. A preplanned interim analysis was done for data accumulated from 81% of the 3-year study exposure. This study was registered with ClinicalTrials.gov, number NCT00666224. FINDINGS: All randomly assigned participants were analysed for the primary outcome. Glatiramer acetate reduced the risk of developing clinically definite multiple sclerosis by 45% compared with placebo (hazard ratio 0.55, 95% CI 0.40-0.77; p=0.0005). The time for 25% of patients to convert to clinically definite disease was prolonged by 115%, from 336 days for placebo to 722 days for glatiramer acetate. The most common adverse events in the glatiramer acetate group were injection-site reactions (135 [56%] glatiramer acetate vs 56 [24%] placebo) and immediate post-injection reactions (47 [19%] vs 12 [5%]). INTERPRETATION: Early treatment with glatiramer acetate is efficacious in delaying conversion to clinically definite multiple sclerosis in patients presenting with clinically isolated syndrome and brain lesions detected by MRI. FUNDING: Teva Pharmaceutical Industries, Israel. PMID- 19815270 TI - Cell recruitment and transfection in gene activated collagen matrix. AB - Realization of systems able to both recruit cells and influence their fate (affecting their processes) represents a new approach for tissue regeneration. We investigated the potency of gene activated matrix (GAM) and implemented the GAM strategy in order to achieve a control of gene expression, as well as a specific cell recruitment. To this aim we developed a 3D DNA bio-activated collagen matrix by Poly (ethylenimine) (PEI)/DNA complex immobilization in the matrix through biotin/avidin bond. Moreover, we realised a serum based chemotactic gradient within the matrix in order to directionally attract NIH3T3 cells. In this system, cells are recruited and forced to migrate through the matrix where they find the bound PEI/DNA complexes and are transfected. The transfected cells can act as local in vivo bioreactors, secreting plasmid encoded proteins that augment tissue repair and regeneration. 3D cell migration and cell transfection were monitored through time-lapse video microscopy and fluorescence microscopy. Cell transfection was also quantified through FACS analysis. Results show that our engineered matrix is able to recruit external cells and transfect them once internalized, therefore it could help in tissue repairing strategy. PMID- 19815271 TI - Alternative bearing materials for intervertebral disc arthroplasty. AB - The objective of our study was to test alternative polymer-on-polymer articulations for cervical total disc arthroplasty with favourable biotribological properties and the benefit of radiolucency in comparison to the clinically well established metal-on-polyethylene coupling. In vitro wear simulation was performed according to ISO 18192-1:2008 (E) with the clinically introduced activ C cervical artificial disc (Aesculap AG Tuttlingen, Germany) made of UHMWPE/CoCr29Mo6 in a direct comparison to experimental disc articulations made of PEEK, CFR-PEEK and PEK. Each material combination was tested for 10 million cycles with a customised 6 station spinal wear simulator (EndoLab Thansau, Germany). Gravimetric and geometric wear assessment, optical surface characterisation and an estimation of particle size and morphology were performed. The gravimetric wear rate of the clinical reference polyethylene-on cobalt-chromium was 1.0+/-0.1 mg/million cycles, compared to 1.4+/-0.4 mg/million cycles for PEEK, to 0.02+/-0.02 mg/million cycles for CFR-PEEK and 0.8+/-0.1 mg/million cycles for PEK. In conclusion, a number of different candidate materials for total cervical disc arthroplasty were compared using the same disc design. Whereas the polymer-on-polymer articulation of PEK showed no substantial benefit in comparison to polyethylene-on-cobalt-chromium and whereas natural PEEK tends towards pitting and delamination, the carbon fibre reinforced PEEK demonstrated an excellent wear behaviour with a reduction in order of a magnitude. Therefore, the CFR-PEEK based polymer-on-polymer articulations may be an alternative to polyethylene-on-metal and have a high potential for next generation disc replacements. PMID- 19815272 TI - The osteogenic differentiation of adult bone marrow and perinatal umbilical mesenchymal stem cells and matrix remodelling in three-dimensional collagen scaffolds. AB - Adult human mesenchymal stem cells from bone marrow (BM-MSC) represent a promising source for skeletal regeneration. Perinatal MSC from Wharton's Jelly of the umbilical cord (UC-MSC) are expected to possess enhanced differentiation capacities due to partial expression of pluripotency markers. For bone tissue engineering, it is important to analyse in vitro behaviour of stem cell/biomaterial hybrids concerning in vivo integration into injured tissue via migration, matrix remodelling and differentiation. This study compares the cell mediated remodelling of three-dimensional collagen I/III gels during osteogenic differentiation of both cell types. When activated through collagen contact and subjected to osteogenic differentiation, UC-MSC differ from BM-MSC in expression and synthesis of extracellular matrix (ECM) proteins as shown by histology, immunohistochemistry, Western Blot analysis and realtime-RT-PCR. The biosynthetic activity was accompanied in both cell types by the ultrastructural appearance of hydroxyapatite/calcium crystals and osteogenic gene induction. Following secretion of matrix metalloproteinases (MMP), both MSC types migrated into and colonised the collagenous matrix causing matrix strengthening and contraction. These results indicate that UC-MSC and BM-MSC display all features needed for effective bone fracture healing. The expression of ECM differs in both cell types considerably, suggesting different mechanisms for bone formation and significant impact for bone tissue engineering. PMID- 19815273 TI - High throughput methods applied in biomaterial development and discovery. AB - The high throughput discovery of new bio materials can be achieved by rapidly screening many different materials synthesised by a combinatorial approach to identify the optimal composition that fulfils a particular biomedical application. Here we review the literature in this area and conclude that for polymers this process is best achieved in a microarray format, which enable thousands of cell-material interactions to be monitored on a single chip. Polymer microarrays can be formed by printing pre-synthesised polymers or by printing monomers onto the chip where on-slide polymerisation is initiated. The surface properties of the material can be analysed and correlated to the biological performance using high throughput surface analysis, including time-of-flight secondary ion mass spectrometry (ToF-SIMS), X-ray photoelectron spectroscopy (XPS) and water contact angle (WCA) measurements. This approach enables the surface properties responsible for the success of a material to be understood, which in turn provides the foundations of future material design. The high throughput discovery of materials using polymer microarrays has been explored for many cell-based applications including the isolation of specific cells from heterogeneous populations, the attachment and differentiation of stem cells and the controlled transfection of cells. Further development of polymerisation techniques and high throughput biological assays amenable to the polymer microarray format will broaden the combinatorial space and biological phenomenon that polymer microarrays can explore, and increase their efficacy. This will, in turn, facilitate the discovery of optimised polymeric materials for many biomaterial applications. PMID- 19815274 TI - Symptomatology in adolescents following initial disclosure of sexual abuse: the roles of crisis support, appraisals and coping. AB - OBJECTIVE: The present study examined possible explanations for symptom development and variability in sexually abused adolescents. A theoretical model of sexual abuse, in which appraisal, coping and crisis support play prominent roles, was tested in a clinical group of 100 sexually abused adolescents. METHOD: Participants, aged 12-18 years, completed questionnaires regarding severity of abuse, negative appraisals, crisis support, coping strategies, and trauma-related stress symptoms. RESULTS: Since severity of abuse had no influence on the association between appraisals and symptoms it was eliminated from our model. Structural equation modeling analyses showed a significant interrelationship between the examined concepts. Adolescents who appraised the abuse as more threatening showed more internalizing and externalizing trauma symptoms. Moreover, more negative appraisals were associated with more avoidance as well as with more active coping strategies. Direct crisis support was associated with less negative appraisals and with the use of more active coping strategies. CONCLUSIONS: The study showed that especially the buffering role of crisis support and the role of negative appraisals explained symptom development and variety in sexually abused adolescents. PRACTICE IMPLICATIONS: Adaptation to sexual abuse includes the ways in which adolescents perceive the event as threatening and harmful, how they cope with the abusive experience, and how they receive direct support from their family. Clinicians, therefore, need to expand the focus of their assessments beyond age- and trauma-specific symptoms, to age- and trauma-specific appraisals, coping, and social support. The assessment of these factors should take place as soon as possible after the disclosure of the abuse. Individualized treatment plans and clinical interventions need to be based on these personal and environmental variables, rather than reliance on the influence of abuse-related characteristics, such as the severity or type of abuse. PMID- 19815275 TI - Murine BAFF-receptor residues 168-175 are essential for optimal CD21/35 expression but dispensable for B cell survival. AB - BAFF-R (B cell-activating factor belonging to the tumor necrosis factor family receptor) regulates B lymphocyte survival, maturation, homeostasis, and self tolerance through signaling mechanisms that are not completely understood. A spontaneous BAFF-R mutation, Bcmd-1, disrupts BAFF-R signaling. However, it is not clear why the Bcmd-1-encoded BAFF-R fails to adequately support B cell survival, optimal CD21/35 expression, and B-cell tolerance to dsDNA, since it is 95% identical to the wild-type (wt) BAFF-R and retains the only known signaling motif. A retrotransposon insertion in A/WySnJ strain mice generated the Bcmd-1 allele, replacing the eight C-terminal BAFF-R residues with 21 retrotransposon encoded residues. New data reported here show that the displaced residues, previously thought to have no signaling role, are essential for optimal CD21/35 expression but contribute little to B cell survival signaling. Analysis of wt Baffr or Bcmd-1 homozygous (A/WySnJ X B6.BCL2)F2 mice confirmed that BCL2 complemented Bcmd-1 for B cell survival but not CD21/35 expression. Through in vivo retroviral transduction experiments, we show that Baffr complemented Bcmd-1 for B cell survival but not CD21/35 expression, whereas the BaffrDelta103-175 deletion mutant lacking the BAFF-R cytoplasmic domain failed to support these functions. Importantly, we show that the BaffrDelta168-175 deletion mutant lacking the retrotransposon-displaced residues, and a BaffrT170A mutant lacking a critical threonine, supported B cell survival but failed to support optimal CD21/35 expression. These data provide the first evidence for a possible bifurcation at the receptor level in the BAFF-R signaling pathway. We suggest that discrete BAFF-R cytoplasmic domains may interact with distinct downstream pathways to provide fine control over B cell survival, maturation, and tolerance induction. PMID- 19815276 TI - Prevalence of visually significant cataract and factors associated with unmet need for cataract surgery: Los Angeles Latino Eye Study. AB - PURPOSE: To estimate the prevalence of visually significant cataract in a US Latino population and to report predisposing, enabling, need, and health behavior characteristics associated with the unmet need for cataract surgery (UNCS). DESIGN: Population-based, cross-sectional study. PARTICIPANTS: A total of 6142 Latinos 40 years and older from 6 census tracts in Los Angeles County, California. METHODS: Participants completed an in-home interview and a comprehensive eye examination that included assessment of lens opacification, using the slit lamp-based Lens Opacities Classification System II (LOCS II), and best-corrected visual acuity. Visually significant cataract was defined by any LOCS II grading >or=2, best-corrected visual acuity <20/40, cataract as the primary cause of vision impairment, and self-reported vision of fair or worse. Because cataract surgery is not needed in all persons, participants with a visually significant cataract or prior cataract surgery in at least 1 eye composed the at-risk cohort needing cataract surgery. Unmet need for cataract surgery was defined as any person in the at-risk cohort who had at least 1 eye with a visually significant cataract. Univariate and stepwise logistic regression analyses were used to identify predisposing, enabling, need, and health behavior characteristics associated with UNCS. MAIN OUTCOME MEASURES: Prevalence of visually significant cataract and odds ratios (ORs) for factors associated with UNCS. RESULTS: Of 6142 participants who completed the interview and clinical examination, 118 (1.92%) had visually significant cataract in at least 1 eye. Of the 344 participants who have needed cataract surgery, 118 (34.3%) had UNCS. Independent factors associated with UNCS included health behavior: having last eye examination >or=5 years ago compared with <1 year ago (OR, 3.76; 95% confidence interval [CI], 1.71-8.25), and enabling factors: being uninsured (OR, 2.79; CI, 1.30-5.19), income less than $20,000 (OR, 2.60; CI, 1.40-5.56), and self-reported barriers to eye care (OR, 2.41; CI, 1.14-5.13). CONCLUSIONS: Latinos in our study had a substantial UNCS. Because Latinos with specific health behavior and enabling characteristics were more likely to have UNCS, interventions aimed at modifying these characteristics may be beneficial in reducing the unmet need and thus reducing the burden of visual impairment related to cataract in the United States. PMID- 19815277 TI - Incidence of dislocation of intraocular lenses and pseudophakodonesis 10 years after cataract surgery. AB - PURPOSE: To estimate the incidence of early and late intraocular lens (IOL) dislocation and the frequency of pseudophakodonesis in a population-based cohort of cataract surgery cases. The patients were followed up from before to 10 years after surgery. DESIGN: Cohort study. PARTICIPANTS: Eight hundred ten cataract surgery patients. METHODS: A prospective population-based cohort of 810 cataract surgery patients with presenile or senile cataracts was examined before surgery. Ten years later, 289 (73%) of 395 survivors agreed to participate in an eye examination. In addition to a routine eye examination of the anterior and posterior segment, all eyes were assessed for pseudophakodonesis and significant dislocation of the IOL. The medical records were studied and information concerning previous postoperative surgical interventions such as IOL exchange or repositioning was noted. This information was also obtained from the records of the deceased patients and those unable or unwilling to participate. The material was analyzed statistically. MAIN OUTCOME MEASURES: Previous IOL exchange or repositioning surgery, significant IOL dislocation, and degree of pseudophakodonesis. RESULTS: Most patients (n = 795/810; 98%) underwent sutureless clear corneal phacoemulsification surgery with a 3.2-mm temporal incision. A foldable IOL was implanted, 95% of which were an Alcon MA60BM AcrySof (Alcon Inc, Fort Worth, TX). Approximately 40% of the patients had pseudoexfoliations (PEX). After a 10-year follow-up, 5 (0.6%) of the 800 patients at risk required surgery for a dislocated IOL. All of these patients were male, and in all cases, the dislocation was late and within the capsular bag. The cumulative incidence over 10 years was 1%. At the examination 10 years after surgery, 2 (0.7%) of 287 patients at risk had pronounced pseudophakodonesis and 4 (1.4%) had moderate pseudophakodonesis. CONCLUSIONS: The 10-year cumulative incidence of dislocated IOLs needing surgical attention was low in this population-based cohort with a high frequency of PEX. Early dislocation did not occur in any of the patients. The risk of this complication in an individual patient seems to be low. Because of the large number of people with previous cataract surgery, dislocated IOLs may cause a relatively large public health care burden. PMID- 19815278 TI - Imaging of long-term retinal damage after resolved cotton wool spots. AB - PURPOSE: Patients infected with the human immunodeficiency virus (HIV) develop noninfectious retinopathy characterized by retinal cotton wool spots (CWS) and microvascular abnormalities. Ophthalmoscopically, CWS fade with time. We hypothesized that structural changes should be permanent and possibly visible well after ophthalmoscopic resolution. We used simultaneous spectral domain optical coherence tomography (SD-OCT)/scanning laser ophthalmoscope (SLO) to allow colocalization of the lesions and determine the extent and location of residual damage after ophthalmoscopic resolution of the lesions. DESIGN: Retrospective, noninterventional case series. PARTICIPANTS: Eight eyes of 7 HIV patients with 19 resolved retinal CWS. METHODS: Nineteen retinal CWS were imaged between 2 and 16 years (median, 7.84) after the acute lesions using simultaneous SD-OCT and SLO examinations. The areas of the previous CWS were scanned by overlaying the color retinal image over the SLO image and scanning at high resolution in the horizontal plane through the resolved lesion. Each CWS lesion had a control area taken from the same eye within 2 disc diameters of the lesion. The thickness of each of the retinal layers was compared between lesions and control areas using a paired t-test with multitest correction. MAIN OUTCOME MEASURES: Thickness of the retinal nerve fiber layer (RNFL), ganglion cell layer (GCL), inner plexiform layer (IPL), inner nuclear layer (INL), outer plexiform layer (OPL), and outer nuclear layer (ONL). RESULTS: The greatest loss of thickness was seen in the retinal GCL with a 43% reduction in thickness. There was a statistically significant thinning of the RNFL, GCL, IPL, INL, and OPL. The median thickness differences ranged from 5 to 7 microns. This difference was highly significant. Another striking finding was the displacement of the ONL toward the retinal surface resulting in an apparent increase in thickness of the ONL by >15% (median difference, 12 microns). CONCLUSIONS: Our data, using ultrahigh resolution and high-speed SD-OCT/SLO, show and quantify the presence of permanent retinal destruction associated with retinal CWS in HIV disease. PMID- 19815279 TI - Major eye diseases and risk factors associated with systemic hypertension in an adult Chinese population: the Beijing Eye Study. AB - PURPOSE: To assess the relationship of hypertension with major eye diseases and other ocular parameters. DESIGN: Population-based study. PARTICIPANTS: The Beijing Eye Study is a population-based study that included 4439 Chinese subjects examined at the baseline examination in 2001; there was a follow-up examination in 2006, in which 3251 subjects participated, of whom 3222 had blood pressure measurements. METHODS: All participants underwent an ophthalmic examination, anthropometric measurements, and blood pressure measurement. Hypertension was defined as a systolic blood pressure >or=140 mmHg and/or diastolic blood pressure >or=90 mmHg, and/or self-reported current treatment for hypertension with antihypertensive medication. MAIN OUTCOME MEASURES: Blood pressure and ocular parameters, including intraocular pressure and prevalence of major ophthalmic diseases. RESULTS: Mean age of participants in the present study was 60.4+/-10.0 years. Hypertension was present in 1500 (46.6%) of the 3222 subjects who had their blood pressure measured. In multiple regression analysis, hypertension was associated with higher intraocular pressure (beta = 0.39; 95% confidence interval [CI], 0.12-0.66; P = 0.005), focal arteriolar narrowing (odds ratio [OR], 1.78; 95% CI, 1.34-2.36; P<0.001), arteriovenous nicking (OR, 1.50; 95% CI, 1.11-2.04; P = 0.009), generalized retinal arteriolar narrowing (OR, 1.65; 95% CI, 1.30 2.09; P<0.001), retinal vein occlusions (OR, 2.86; 95% CI, 1.21-6.80; P = 0.02), and diabetic retinopathy (OR, 1.90; 95% CI, 1.08-3.31; P = 0.02). Hypertension was not significantly associated with the prevalence of open-angle glaucoma (P = 0.19), angle-closure glaucoma (P = 0.15), age-related macular degeneration (AMD) (P = 0.73), nuclear cataract (P = 0.88), posterior subcapsular cataract (P = 0.30), cortical cataract (P = 0.10), or area of alpha zone (P = 0.05) or beta zone of parapapillary atrophy (P = 0.95). CONCLUSIONS: In Chinese persons, while controlling for other systemic parameters, hypertension was associated with increased intraocular pressure, retinal microvascular abnormalities, and prevalence of retinal vein occlusion and diabetic retinopathy. Hypertension was not associated significantly with AMD, age-related cataract, or glaucoma. PMID- 19815280 TI - Reticular pseudodrusen are subretinal drusenoid deposits. AB - PURPOSE: To characterize reticular pseudodrusen, a potential risk factor for late age-related macular degeneration. DESIGN: Retrospective, observational case series. PARTICIPANTS: Fifty-eight eyes of 33 patients with pseudodrusen (20 female). METHODS: Consecutive patients with reticular pseudodrusen, diagnosed by their typical appearance and distribution using ophthalmoscopy, the blue channel of color fundus photographs, and near infrared images. The patients were imaged by spectral domain optical coherence tomography (SD OCT), and correlations were made between the near infrared images and the SD OCT images. The SD OCT findings in patients with pseudodrusen were compared with previously reported histologic findings of subretinal drusenoid deposits. The histologic specimens were reevaluated with the additional knowledge of the clinical information. MAIN OUTCOME MEASURES: Spectral domain optical coherence tomography and histologic characteristics of pseudodrusen. RESULTS: The mean age of the 33 patients was 81.7 years. The correlating SD OCT scans showed collections of granular hyperreflective material above the retinal pigment epithelium (RPE), in the subretinal space located primarily between the RPE and the boundary between the inner and outer segments of the photoreceptors (IS/OS boundary). In a more advanced stage, this material formed small mounds that broke through the IS/OS boundary. There were no correlates to the deposits seen under the RPE or in the choroid. These findings were similar in character to previously reported histologic characterization of subretinal drusenoid deposits, which had identified the presence of membranous debris, unesterified cholesterol, and complement within the deposits. CONCLUSIONS: Pseudodrusen seen by clinical examination may be subretinal drusenoid deposits seen by histologic examination. This unexpected location suggests that potential pathophysiologic mechanisms on both sides of the RPE need to be taken into account in theories related to the development of age-related macular degeneration. PMID- 19815281 TI - Intraocular lens power selection in the second eye of patients undergoing bilateral, sequential cataract extraction. AB - PURPOSE: To determine if the 1-month postoperative error of predicted refraction of the first eye can be used to alter intraocular lens (IOL) power selection and improve refractive results for the second eye in patients undergoing bilateral, sequential phacoemulsification with IOL implantation (phaco/IOL). DESIGN: Retrospective, consecutive, case series. PARTICIPANTS: Three hundred consecutive patients who underwent uncomplicated bilateral, sequential phaco/IOL between January 1, 2006, and December 31, 2007, by a single surgeon using a single IOL platform. METHODS: The observed second eye 1-month postoperative spherical equivalent refractive error was compared with calculations of the hypothetic 1 month postoperative spherical equivalent refractive error if the first eye error had been fully or partially incorporated into the choice of IOL power for the second eye. The optimal amount of partial adjustment was determined. MAIN OUTCOME MEASURES: The error of predicted refraction: the difference between the actual or hypothetic spherical equivalent refractive errors and those predicted by preoperative calculations. RESULTS: A total of 206 patients met inclusion criteria. The average of the absolute value of the unadjusted second eye error (|E(UNADJ)|) was +0.44 diopters (D) compared with +0.42 D for the fully adjusted second eye error (|E(FULL)|). The optimal amount of adjustment of the second eye IOL power was determined to be 50%; the average of the absolute value of this partially adjusted second eye error (|E(PARTIAL,50%)|) was +0.36 D; this was statistically different from |E(UNADJ)| (P<0.0001) and |E(FULL)| (P = 0.001). The statistically significant benefit was observed for patients with either myopic or hyperopic errors in the first eye. The percentages of patients achieving postoperative refractions within 0.5 D and 1.0 D of the predicted refraction were 66.5% and 90.3%, respectively, for the unadjusted second eye, 67.0% and 90.8%, respectively, for the hypothetic fully adjusted second eye, and 74.3% and 93.7%, respectively, for the hypothetic partially adjusted (50%) second eye. CONCLUSIONS: Accounting for 50% of the observed error of predicted refraction of the first eye reduced the error of predicted refraction in the second eye. This novel methodology has the potential to improve the refractive outcomes in the second eye of patients with cataract. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article. PMID- 19815282 TI - An outbreak of post-cataract surgery endophthalmitis caused by Pseudomonas aeruginosa. AB - OBJECTIVE: Infectious endophthalmitis is among the most serious complications of cataract surgery. Gram-negative bacteria, including Pseudomonas aeruginosa, are responsible for less than 30% of cases; however, their rapidity of infection and virulence often results in poor visual outcome despite prompt antibiotic treatment. The purpose of this study was to investigate an outbreak of post cataract surgery P. aeruginosa endophthalmitis in India. DESIGN: Hospital-based case series. PARTICIPANTS: Twenty patients with acute postoperative endophthalmitis who underwent cataract surgery at one of the peripheral centers of Joseph Eye Hospital, Tiruchirapalli, Tamil Nadu, India, from February 23 to April 2, 2008. INTERVENTIONS: Vitreous aspirates and environmental surveillance specimens were inoculated for culture. Antibiotic susceptibility testing was performed by agar diffusion method. Polymerase chain reaction (PCR) with enterobacterial repetitive intergenic consensus (ERIC) primers (ERIC-PCR) was used to establish the clonal relationship between clinical and environmental isolates. MAIN OUTCOME MEASURES: Post-cataract surgery P. aeruginosa endophthalmitis. RESULTS: Pseudomonas aeruginosa was isolated from 20 eyes with postoperative endophthalmitis, the phacoemulsifier's internal tubes, the povidone iodine solution, and the operating theater air-conditioning system. All strains were multidrug-resistant to cefazolin, chloramphenicol, tetracycline, aminoglycosides, and fluoroquinolones; conversely, most of them were susceptible to polymyxin B. Polymerase chain reaction with enterobacterial repetitive intergenic consensus primers disclosed 2 major clusters: six genetically identical clinical isolates shared 94% of similarity with the air-conditioning isolate; 11 other clinical isolates had 88% of similarity with the former strain. Despite the prompt use of intravitreal antibiotics, 10 patients had evisceration or phthisis of the affected eye. CONCLUSIONS: The outcome of cataract surgery related P. aeruginosa endophthalmitis is poor. The detection of multidrug resistant isolates is a serious problem, jeopardizing an appropriate choice of treatment. Polymerase chain reaction with enterobacterial repetitive intergenic consensus results strongly suggest that the main source of infection in this outbreak was the contaminated air-conditioning system. Polymerase chain reaction with enterobacterial repetitive intergenic consensus is an inexpensive, fast, reproducible, and discriminatory DNA typing tool for effective epidemiologic surveillance of clinical and environmental isolates of P. aeruginosa. PMID- 19815283 TI - Analysis of diluted vitreous samples from vitrectomy is useful in eyes with severe acute postoperative endophthalmitis. AB - PURPOSE: This study was designed to compare the diagnostic yield of microbiological analysis performed on diluted and undiluted vitreous samples from pars plana vitrectomy (PPV) in patients with acute postcataract endophthalmitis. DESIGN: Cohort study, evaluation of diagnostic test or technology. PARTICIPANTS: Patients with acute postcataract endophthalmitis (<6 weeks). METHODS: Undiluted and diluted vitreous samples were taken from 34 consecutive patients at the beginning of PPV as part of the multicenter prospective study of the French Institutional Endophthalmitis Study (FRIENDS) group. Vitrectomy was performed after 1 (n = 12) or 2 (n = 22) intravitreous antibiotic injections. McNemar's nonparametric test was used to compare culture and polymerase chain reaction (PCR) results between diluted and undiluted samples. MAIN OUTCOME MEASURES: Rate of positivity of conventional culture (brain heart infusion broth) and eubacterial PCR tests from undiluted and diluted vitreous samples. RESULTS: The microbiological analysis of both undiluted and diluted vitreous samples detected and identified a bacterial pathogen in 26 out of 34 cases (76.4%). The analysis of undiluted and diluted vitreous at the time of PPV, using eubacterial PCR and conventional culture, gave similar results (P = 0.99; McNemar test). However, eubacterial PCR was more sensitive than culture in detecting bacteria in vitreous at the time of PPV (76% vs 6%; P = 0.001; McNemar test). The difference in sensitivity between the 2 techniques was primarily associated with false-negative culture results for undiluted samples (2/3 of cases), mainly for coagulase negative staphylococci. CONCLUSIONS: The microbiological results obtained combining PCR and culture techniques were similar for diluted vitreous and undiluted vitreous analysis. When eubacterial PCR is available, sampling diluted vitreous, an easier procedure, may replace sampling undiluted vitreous. PMID- 19815284 TI - Long-term topical steroid treatment after penetrating keratoplasty in patients with pseudophakic bullous keratopathy. AB - PURPOSE: To assess the use of long-term topical corticosteroid treatment in patients with pseudophakic bullous keratopathy (PBK) after penetrating keratoplasty (PK). DESIGN: Retrospective cohort study. PARTICIPANTS: This study considered patients with PBK undergoing an initial PK procedure for visual reasons in the United Kingdom between April 1999 and March 2004. There were 1274 initial PK procedures for PBK reported to United Kingdom Transplant in this period, of which 1184 (91%) were grafted for visual reasons. Of these 1184 grafts, follow-up was reported in 1033 instances (87%). METHODS: A Cox regression model was used to investigate the combined effects of all preoperative factors (recipient age, human leukocyte antigen [HLA] matching, trephine size, deep stromal vascularization, surgeon activity) on graft failure. The model was fitted using all preoperative factors first, and subsequently, factors associated with corticosteroid and other medications were included. MAIN OUTCOME MEASURES: Graft survival. RESULTS: Three-year survival of grafts for PBK was 65% (95% confidence interval [CI], 59%-70%). Topical corticosteroids were still being used beyond 18 months after surgery in 378 (37%) of the 1033 corneal grafts included in this study. The grafts of patients not currently receiving steroids were 1.5 times as likely to fail (hazard ratio [HR], 1.5; 95% CI, 1.0-2.2; P<0.03). Lack of HLA matching (P = 0.006), trephine size or=8.00 mm (P = 0.03), recipient age younger than 65 years (P = 0.003), and corneal vascularization (P = 0.04) all increased the risk of graft failure. CONCLUSIONS: The use of long-term postoperative corticosteroids improved graft survival after PK for PBK. Barring patient contraindications for long-term topical corticosteroid use, clinicians should consider maintaining patients with PBK on long-term postoperative corticosteroid maintenance. PMID- 19815285 TI - Predicted long-term outcome of corneal transplantation. AB - OBJECTIVE: To analyze graft survival and the outcome of the corneal endothelium after corneal transplantation in a single model to predict the long-term prognosis of these grafts. DESIGN: Cohort study. Data were recorded prospectively and then analyzed retrospectively. PARTICIPANTS: One thousand one hundred forty four consecutive eyes of 1144 patients who underwent corneal transplantation between 1992 and 2006. INTERVENTIONS: Penetrating keratoplasty and deep anterior lamellar keratoplasty. MAIN OUTCOME MEASURES: Slit-lamp examination and wide field specular microscopy results. A joint analysis of endothelial cell loss and time to graft failure was undertaken. From midterm simultaneous analysis of graft survival and endothelial cell loss, long-term graft survival was predicted. RESULTS: The observed 5- and 10-year graft survival estimates were, respectively, 74% and 64%. The average endothelial cell density (cell loss) was 2270 cells/mm(2) before surgery, 1058 cells/mm(2) (-53%) during the sixth postoperative year, and 865 cells/mm(2) (-61%) during the 10th postoperative year. Overall, the predicted graft survival estimate was 27% at 20 years and 2% at 30 years. Both observed and predicted graft survival were higher in patients who had undergone lamellar keratoplasty than in patients who had undergone penetrating keratoplasty and had normal recipient endothelium and higher in patients who had undergone penetrating keratoplasty and had normal recipient endothelium than in patients who had undergone penetrating keratoplasty and had impaired recipient endothelium. CONCLUSIONS: For corneal diseases involving the endothelium, penetrating keratoplasty seems to be a good therapeutic approach in elderly patients because the graft life-span may be similar to the patient life expectancy. Conversely, for younger patients, penetrating keratoplasty is only a midterm therapeutic approach. For corneal diseases not involving the endothelium, deep anterior lamellar keratoplasty seems to be a promising therapeutic approach with higher long-term expected survival. PMID- 19815286 TI - Interventions improve poor adherence with once daily glaucoma medications in electronically monitored patients. AB - PURPOSE: To investigate the impact of an intervention program to improve adherence with topical, once daily therapy for glaucoma. DESIGN: Randomized controlled clinical trial. PARTICIPANTS: Sixty-six patients with glaucoma being treated with a prostaglandin analog in 1 or both eyes at the Scheie Eye Institute or Wilmer Eye Institute between November 2006 and June 2007. METHODS: In an observational study, participants who took 75% or fewer doses (as measured using the travoprost Dosing Aid [DA]) during an initial 3-month period were randomized into 2 groups. The intervention group watched an educational video, reviewed current barriers to drop-taking and possible solutions with a study coordinator, received regular phone call reminders, and had audible and visible reminders activated on their DA devices. The control group was told to take drops as prescribed and received no additional intervention. MAIN OUTCOME MEASURES: Change in drop use adherence as determined by the DA device. RESULTS: In the 3-month observation period before randomization, intervention group patients had used a mean of 54+/-17% of scheduled doses, and this increased to 73+/-22% during the following 3-month period (P<0.001, n = 35). The control mean adherence rate of 46+/-23% at baseline was statistically unchanged during the follow-up observation period (51+/-30%, P = 0.16, n = 31). In a multivariate analysis, intervention, baseline compliance rate of <50%, and white ethnicity were predictors of improved adherence during the 3 months of intervention. The intraocular pressure (IOP) of the intervention and control groups did not change between months 3 and 6 after intervention (P = 0.96, 0.34, respectively), and there was no correlation of IOP change with adherence rate change between both groups (Pearson correlation r = 0.06, P = 0.51). CONCLUSIONS: A multifaceted intervention significantly increased adherence with glaucoma medications. Those with improved adherence were in the intervention group, had very low adherence rates at baseline, and were white. IOP did not correlate with adherence. Further research is needed to determine which components of this intervention were most effective. PMID- 19815287 TI - Emerging prevalence of microsporidial keratitis in Singapore: epidemiology, clinical features, and management. AB - OBJECTIVE: To investigate the incidence and epidemiologic factors involved in the development of microsporidial keratitis. The association of host immune status and clinical pattern, clinical features, and the role of fluoroquinolone monotherapy in treatment are also examined. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: All cases (124 patients, 134 eyes) of microsporidial keratitis confirmed with modified trichrome stain positive of corneal scrape over a 4-year period. METHODS: Epidemiologic factors were observed. Host immune status with human immunodeficiency virus (HIV) serology and CD4/CD8 analysis was performed when consent was obtained. Visual acuity (VA) and slit-lamp examination throughout the course of keratitis was recorded. Treatment used included topical fluoroquinolones (ciprofloxacin 0.3%, moxifloxacin 0.5%, gatifloxacin 0.5%, levofloxacin 0.5%, or norfloxacin 0.3%) as monotherapy or in combination with topical fumagillin and/or systemic albendazole. Where corneal edema developed, ultrasound corneal pachymetry was recorded. MAIN OUTCOME MEASURES: Demographic features and epidemiologic factors, including host immune status. Clinical features and disease course, including the response to different therapeutic regimes. RESULTS: Patients ranged in age from 11 to 68 years (mean, 31.9; median, 30) with a male:female ratio of 8:1 (females n = 17 [13.7%]). We performed HIV serology and CD4/CD8 in 45.9% of cases (n = 57); all the cases tested were negative with normal T-cell indices. Epidemiologic factors included soil exposure (50%), contact lens wear (21.1%), and topical steroid treatment (17.1%). The VA on presentation ranged from 20/20 to 20/100 (median, 20/30) with no loss in lines of VA on resolution. Common features were follicular papillary conjunctivitis and coarse punctate epithelial lesions in 3 patterns--diffuse, peripheral, and paracentral--evolving into nummular keratitis before resolution. Resolution occurred in 99% of cases on topical fluoroquinolone monotherapy. Four patients had recurrent disease that resolved with repeat fluoroquinolone or fluoroquinolone/oral albendazole combination. Two new clinical features were identified--diffuse endotheliitis (19.4%) with corneal edema and limbitis. CONCLUSIONS: This study identifies an increasing incidence of microsporidial keratitis in Singapore with a strong correlation with prior soil exposure. Diffuse endotheliitis and limbitis have not been described and resolves with topical steroid therapy. Topical fluoroquinolone monotherapy is a valid treatment option. PMID- 19815288 TI - Uveitis in a patient treated with Bacille-Calmette-Guerin: possible antigenic mimicry of mycobacterial and retinal antigens. AB - PURPOSE: To investigate the cellular immune response in uveitis developing after intravesical Bacille-Calmette-Guerin (BCG) applications. DESIGN: Experimental study. PARTICIPANTS: A 72-year-old HLA-B27-negative patient with bilateral granulomatous anterior uveitis that developed during the third cycle of intravesical BCG applications she was receiving for treatment of bladder carcinoma. METHODS: The patient's peripheral T cell reactivity to ocular autoantigens was compared with the response to purified protein derivative (PPD) from Mycobacterium tuberculosis. T-cell proliferation and cytokine and chemokine secretion were measured in vitro. MAIN OUTCOME MEASURES: Anterior uveitis was treated successfully with topical corticosteroids and cycloplegics. RESULTS: The following were demonstrated: proliferation to PPD, interphotoreceptor retinoid binding protein (IRBP), and IRBP-peptide R16, as well as secretion of proinflammatory cytokines in response to PPD, retinal soluble antigen (S-Ag), IRBP, cellular retinal-binding protein (CRALBP), and some S-Ag and IRBP peptides. CONCLUSIONS: These data indicate the generation of a polyclonal autoimmune reaction elicited by BCG. Amino acid sequence alignments revealed homologies between proteins from M. tuberculosis, BCG, and retinal antigens, suggesting antigenic mimicry as a potential cause of uveitis in this patient. PMID- 19815289 TI - Optical coherence tomography: pathology correlation of optic disc melanocytoma. AB - OBJECTIVE: To correlate combined optical coherence tomography (OCT) and scanning laser ophthalmoscope (SLO) imaging to clinical and histopathologic characteristics of optic disc melanocytoma (ODM). DESIGN: Retrospective, consecutive clinical case analysis. PARTICIPANTS: Twenty patients with ODM were evaluated. TESTING: All underwent ophthalmic examinations including a best corrected visual acuity, slit-lamp examination, dilated ophthalmoscopy, fundus photography with angiography, B-scan ultrasonography, and combined OCT and SLO imaging. MAIN OUTCOME MEASURES: One eye, treated by enucleation, was evaluated by pathologic analysis and was correlated to its combined OCT and SLO images. These findings were correlated to clinical and OCT information from 19 patients with ODM. RESULTS: Histopathologic and combined OCT and SLO features included: disruption of the internal limiting membrane, disorganization of the retina (overlying the tumor), and visualization of the tumor's subretinal surface. Compressive and infiltrative changes in the optic nerve, peripapillary choroid, and retina were correlated directly to combined OCT and SLO images. To varying degrees, these histopathologic findings were seen on combined OCT and SLO. However, the pigment of the ODMs blocked OCT reflectance deep to the tumor's surface. CONCLUSIONS: This study demonstrated that OCT and SLO findings could be correlated directly with histopathologic characteristics. Imaging with OCT can be used to diagnose and correlate secondary retinal, superficial, and subretinal tumor characteristics. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article. PMID- 19815290 TI - Quantitative iris parameters and association with narrow angles. AB - PURPOSE: To investigate the relationship between quantitative iris parameters (iris curvature [I-Curv], iris area [I-Area], and iris thickness) and the presence of narrow angles. DESIGN: Cross-sectional, community-based study. PARTICIPANTS: We recruited 2047 subjects >50 years old without ophthalmic symptoms from a community clinic in Singapore. METHODS: All subjects underwent gonioscopy and anterior segment optical coherence tomography (AS-OCT) under dark conditions. An eye was considered to have narrow angles if the posterior pigmented trabecular meshwork was not visible for >/=180 degrees on nonindentation gonioscopy with the eye in the primary position. Customized software was used on horizontal AS-OCT scans to measure I-Curv, I-Area, and iris thickness 750 mum (IT750) and 2000 mum (IT2000) from the scleral spur. The average of both temporal and nasal measured values of the right eye was used for analysis. MAIN OUTCOME MEASURES: The association between iris parameters and narrow angles on gonioscopy. RESULTS: Iris parameters from 1465 eyes (71.6%) were available for analysis. Of these, 315 subjects (21.5%) had narrow angles. The mean I-Curv (0.366 vs 0.259 mm; P<0.020), IT750 (0.476 vs 0.453 mm; P<0.001), and IT2000 (0.491 vs 0.482 mm; P = 0.010) were greater in persons with than without narrow angles. After adjusting for age, gender, anterior chamber depth, axial length, and pupil size, the greater I-Curv, I-Area, IT750, and IT2000 were significantly associated with narrow angles (odds ratio [OR] 2.5 and 95% confidence interval [CI], 1.3-5.1; OR, 2.7 and 95% CI, 1.6-4.8; OR, 2.6 and 95% CI, 1.6-4.1; OR, 2.7 and 95% CI, 1.5-4.7, comparing 4th with 1st quartile for each parameter, respectively). In stratified analysis, women and subjects aged >/=60 years had stronger associations for most iris parameters with narrow angles than men and younger subjects. CONCLUSIONS: Quantitative iris parameters (I-Curv, I-Area, and iris thickness) are independently associated with narrow angles, particularly in women and older subjects. These data provide further insights into the pathogenesis of angle closure in Singaporeans. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references. PMID- 19815291 TI - Background comparison of typical age-related macular degeneration and polypoidal choroidal vasculopathy in Japanese patients. AB - OBJECTIVE: To compare background factors of the 2 most dominant subtypes of exudative age-related macular degeneration (AMD) in the Japanese population: typical AMD and polypoidal choroidal vasculopathy (PCV). DESIGN: Cross-sectional comparison. PARTICIPANTS: Consecutive patients with typical AMD (n = 89) and PCV (n = 138) for the primary survey. For the secondary survey, the number of participants was extended to include 148 typical AMD and 170 PCV patients. All the patients included in the present study had been followed up at The University of Tokyo Hospital outpatient macular clinic. METHODS: Background data on gender; age; body mass index; smoking; alcohol consumption; and histories of hypertension, diabetes mellitus (DM), hyperlipidemia, ischemic heart disease, stroke, intensive light exposure, central serous chorioretinopathy (CSC), cataract surgery, glaucoma, and steroid use were obtained mainly through interview. The interviewers were masked to the subtype diagnosis of AMD. Univariate and multivariate logistic regression analyses were performed to identify differences in the background factors between typical AMD and PCV. In the secondary survey, the association of a history of CSC and PCV was confirmed further, and funduscopic findings of an atrophic retinal pigment epithelial (RPE) tract and focal photocoagulation scars that could indicate a history of CSC were investigated. MAIN OUTCOME MEASURES: Frequency and mean of background factors in patients with typical AMD or PCV. RESULTS: The 2 groups showed similar backgrounds with the exception of their histories of DM and CSC. A history of DM was more frequent in typical AMD (24.7% vs. 13.0% in the primary survey; P = 0.027), whereas a history of CSC was more prevalent in PCV (3.4% vs. 14.7% in the secondary survey; P = 0.0005). Funduscopic findings of an atrophic RPE tract or focal photocoagulation scars were found more frequently in PCV (0.7% vs. 7.6%; P = 0.002). CONCLUSIONS: Background factors of typical AMD and PCV are similar but not identical. A history of DM and CSC are more frequent in typical AMD and PCV, respectively. PMID- 19815292 TI - Neovascular age-related macular degeneration: intraocular cytokines and growth factors and the influence of therapy with ranibizumab. AB - PURPOSE: To investigate concentrations of growth factors and inflammatory cytokines in eyes with neovascular age-related macular degeneration (AMD) before and during therapy with intravitreal ranibizumab and to identify associations with disease activity. DESIGN: Prospective clinical trial. PARTICIPANTS AND CONTROLS: Twenty-eight eyes of patients with neovascular AMD were compared with 28 eyes of age-matched patients with cataract as control. METHODS: Ranibizumab was administered intravitreously once at baseline, and retreatments were given at monthly visits if optical coherence tomography (OCT) revealed macular edema or vision loss had occurred. Aqueous humor samples were taken each time intravitreal interventions were performed. Follow-up was 12 months. Luminex (Luminex Inc., Austin, TX) multiplex assays were used for measurement of 29 different growth factors and cytokines, including vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF). MAIN OUTCOME MEASURES: Differences in the concentrations of growth factors and inflammatory cytokines in eyes with neovascular AMD compared with control eyes and the influence of therapy with intravitreal ranibizumab. RESULTS: A significantly increased expression of VEGF (P = 0.033) and a significantly decreased expression of PDGF (P = 0.038) were measured in the aqueous humor of eyes with neovascular AMD. Furthermore, a significant decrease of VEGF (P<0.001) was observed after intravitreal injection of ranibizumab along with significant changes in visual acuity and central retinal thickness (P = 0.039 and P<0.001). During follow-up with a flexible regimen, a correlation was identified between increased VEGF levels and persistent or recurrent macular edema. Changes in PDGF levels were strongly associated with alterations in VEGF concentration. CONCLUSIONS: Vascular endothelial growth factor and PDGF-AA seemed to be associated with disease activity of neovascular AMD. Intravitreal anti-angiogenic treatment with ranibizumab resulted in significantly decreased intraocular VEGF expression below physiologic levels compared with controls. This effect was measurable as long as 4 weeks after each injection and was prolonged by consecutive retreatment. With recurrence after discontinuation of treatment, VEGF levels increased again. PMID- 19815293 TI - Low-dose aspirin and medical record-confirmed age-related macular degeneration in a randomized trial of women. AB - OBJECTIVE: To test whether alternate-day low-dose aspirin affects incidence of age-related macular degeneration (AMD) in a large-scale randomized trial of women. DESIGN: Randomized, double-masked, placebo-controlled trial. PARTICIPANTS: Thirty-nine thousand eight hundred seventy-six healthy female health professionals aged 45 years or older. INTERVENTION: Participants were assigned randomly to receive either 100 mg aspirin on alternate days or placebo and were followed up for the presence of AMD for an average of 10 years. MAIN OUTCOME MEASURES: Incident AMD responsible for a reduction in best-corrected visual acuity to 20/30 or worse based on self-report confirmed by medical record review. RESULTS: After 10 years of treatment and follow-up, there were 111 cases of AMD in the aspirin group and 134 cases in the placebo group (hazard ratio, 0.82; 95% confidence interval, 0.64-1.06). CONCLUSIONS: In a large-scale randomized trial of female health professionals with 10 years of treatment and follow-up, low-dose aspirin had no large beneficial or harmful effect on risk of AMD. PMID- 19815294 TI - Familial asymptomatic macular telangiectasia type 2. AB - OBJECTIVE: To report findings in asymptomatic family members of patients with macular telangiectasia type 2. DESIGN: Prospective, observational, cross sectional case series. PARTICIPANTS: Four patients with symptomatic macular telangiectasia type 2 (index patients) and 5 relatives, including 2 sets of monozygotic twins. METHODS: Screening of family members of participants in a non interventional natural history study of macular telangiectasia type 2. Ophthalmologic examination included best-corrected visual acuity testing, fundus biomicroscopy, fluorescein angiography (FA), optical coherence tomography (OCT), and fundus autofluorescence (FAF) imaging. MAIN OUTCOME MEASURES: Evidence for macular telangiectasia type 2 in any of the imaging methods used and visual function of the family members studied. RESULTS: In the first family, 2 of 3 daughters of a severely affected 68-year-old woman had features of macular telangiectasia type 2. Although one of the daughters was diagnosed by biomicroscopic examination, the second daughter was diagnosed only by subtle changes on OCT and FAF imaging. Both affected daughters were asymptomatic and were unaware that they had the condition. In the second family, clinical examination showed that the 60-year-old brother of the 75-year-old index patient obviously was affected, despite a lack of any subjective visual dysfunction. The 65-year-old monozygotic twin of the third index patient showed a slight retinal thinning within a small area temporal to the foveola in both eyes as well as minor staining on FA and a subtle monocular loss of macular pigment. The 56-year old asymptomatic monozygotic twin of the last proband had opacification of the retina with leakage on FA in the right eye. The fellow eye was unremarkable except for an abnormal FAF signal that was present in both eyes. CONCLUSIONS: Macular telangiectasia type 2 may be more common than previously assumed, but patients may not seek ophthalmic care if their visual function is normal. The study of these early, asymptomatic cases may yield valuable insights into the pathogenesis of the condition. Further research is warranted to determine whether there is an underlying, dominantly inherited genetic abnormality in macular telangiectasia type 2 of variable penetrance and expressivity. PMID- 19815295 TI - Psychoeducation for patients with bipolar disorder receiving lithium: short and long term impact on locus of control and knowledge about lithium. AB - BACKGROUND: Psychoeducation is now considered as part of the integrated treatment for bipolar disorder. But the psychological changes involved in it have been poorly studied. METHOD: We compared the locus of control (LOC, a key variable for health-related behaviours as well as for education practices), the knowledge about lithium [Lithium Knowledge Questionnaire (LKQ)] and attitude about lithium [Attitude towards Lithium Questionnaire (ALQ)] before and after a brief hospital based psychoeducational programme for euthymic patients with bipolar disorder (ICD-10 criteria) receiving lithium prophylaxis. The occurrence of hospitalisations was recorded during the two years before and after the patients underwent psychoeducation. All patients were reassessed after 24 months. RESULTS: 50 consecutive participants at a psychoeducational programme were evaluated. The LKQ but not the ALQ scores increased significantly after the programme. The external "powerful others" component of the LOC significantly increased after psychoeducation. The observed changes were maintained after 24 months. The patients' level of satisfaction was excellent and sustained. There was only a trend for a decrease in the rate of hospitalisations. LIMITATIONS: The knowledge about lithium was assessed with an experimental instrument. Patients followed in a university department may not be representative of bipolar patients at large. CONCLUSIONS: Psychoeducation enduringly increases the knowledge about lithium and induces long term changes in the locus of control that may reflect a shift in illness representations. The LOC may be an important target of psychoeducation for euthymic patients with bipolar disorder. PMID- 19815296 TI - Neurological considerations: autism and Parkinson's disease. AB - Within the spectrum of disorders that manifest obsessive-compulsive (OC) features lies a sub-cluster of neurological conditions. Autism and Parkinson's disease (PD) are examples of two such neurological disorders that seem quite dissimilar on the surface. Yet, both conditions can include repetitive behaviors of a compulsive-impulsive nature. Furthermore, while autism and PD differ in other associated symptom domains that shape the course of each disorder, both disorders share some phenomenology in the core domain of repetitive behaviors and involve basal ganglia and frontal lobe dysfunction, similar to OC disorder (OCD). Accordingly, examination of the similarities and differences between autism and PD may provide insight into the pathophysiology and treatment of OC spectrum disorders. The current review focuses on the phenomenology, comorbidity, course of illness, family history, brain circuitry, and treatment of autism and PD, as they relate to OCD and OC spectrum disturbances. PMID- 19815297 TI - Validation of immunoassays for the candidate renal markers C-reactive protein, immunoglobulin G, thromboxane B2 and retinol binding protein in canine urine. AB - The study of early markers for glomerular and tubular dysfunction in dogs with renal diseases holds promise to gain new insights in the pathogenesis of canine nephropathies. However, the validation of such markers in canine urine is largely lacking. Therefore, immunoassays for the quantification of a set of four urinary markers, C-reactive protein (CRP), immunoglobulin G (IgG), thromboxane B(2) (TXB(2)) and retinol binding protein (RBP), were validated by determining their sensitivity, reproducibility, precision and accuracy in a large patient group. The results show that the immunoassays are appropriate for analysis of urinary CRP, IgG, TXB(2) and RBP in dogs. Furthermore, the significant differences in urinary concentrations of the selected glomerular and tubular markers between healthy (H) dogs and dogs with several types of nephropathies (R) support their future application in both clinical settings and research models. PMID- 19815298 TI - Comparative chemiluminescence of neonatal and adult ovine polymorphonuclear leukocytes. AB - Polymorphonuclear leukocytes (PMN) serve on the first line of the immunological defense of ruminants. However, despite the high morbidity and mortality of neonatal lambs to a wide range of infections there have been no definitive studies undertaken to ascertain whether there might be functional differences in PMN from neonatal lambs when compared to those from adult sheep. To determine whether there were differences in the oxidative respiratory burst of PMN, luminol dependent chemiluminescence (CL) measurements were made of PMN from lambs at 1 week, 1, 2 and 3 months of age and at the same time from their respective dams. PMN isolated from lambs exhibited significantly lower levels of CL until 2 months of age. At 3 months of age the PMN produced levels of CL equal to that of their dams. As CL mirrors the ability of PMN to efficiently phagocytize and kill pathogens the present findings would suggest that PMN of neonatal lambs during the first 2 months are likely less capable of defending them from pathogenic organisms. PMID- 19815299 TI - Brevicompanine E reduces lipopolysaccharide-induced production of proinflammatory cytokines and enzymes in microglia by inhibiting activation of activator protein 1 and nuclear factor-kappaB. AB - Excessive release of proinflammatory cytokines by activated microglia can cause neurotoxicity in neurodegenerative diseases. We found that Brevicompanine E (BE), isolated from a deep ocean sediment derived fungus Penicillium sp., inhibited lipopolysaccharide (LPS)-induced tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), inducible nitric oxide (iNOS) and cyclooxygenase-2 (COX-2) production in microglia. Moreover, electrophoretic mobility shift assay (EMSA) demonstrated that BE attenuated nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1) DNA binding activity in LPS-induced microglia. Consistent with this finding, BE inhibited LPS-induced IkappaBalpha degradation, NF-kappaB nuclear translocation, and also Akt, c-Jun NH2-terminal kinase (JNK) phosphorylation. Thus, BE may be potentially useful for modulating neuroinflammation. PMID- 19815300 TI - Rational and systematic protein purification process development: the next generation. AB - Current biopharmaceutical manufacturing strongly relies on using purification platform processes, offering harmonization of practices and speed-to-market. However, the ability of such processes to respond quickly to anticipated higher quality and capacity demands is under question. Here, we describe novel approaches for purification process development that incorporate biothermodynamics, modern high throughput experimentation and simulation tools. Such development leads to production platform-specific databases containing thermodynamic protein descriptors of major host cell proteins over a range of experimental conditions. This will pave the way for in silico purification process development, providing better process understanding and the potential to respond quickly to product quality and market demands. Future efforts will focus on improving this field further and enabling more rationale in process development. PMID- 19815301 TI - Intra-renal hemodynamics and carotid intima-media thickness in the metabolic syndrome. AB - AIMS: Metabolic syndrome (MetS) is associated with increased cardiovascular risk. We hypothesize that early vascular changes are already present at the time of diagnosis of MetS. The relationship of different measures of early vascular impairment with body fat distribution and the natural progression of MetS was examined in newly diagnosed subjects non-pharmacologically treated. METHODS: 246 consecutively enrolled subjects were categorized according to the presence of MetS and type 2 diabetes (T2D). Intra-renal Doppler flow was used to ascertain resistive (RI) and pulsatility (PI) indices as markers of vascular resistance. Carotid intima-media thickness (IMT), cutis-rectis (CR) and rectis-aorta (RA) thicknesses were measured by ultrasonography; RA/CR ratio was used as measure of body fat distribution. Pro-inflammatory cytokines, C-reactive protein, oxidative markers insulin and adiponectin blood concentrations were also measured. RESULTS: Baseline characteristics demonstrated increasing trends in biochemical, inflammatory, and oxidative parameters from MetS-, MetS+, to MetS+/T2D (p<0.001). After adjusting for age, the same increasing trends across the groups were observed in both sexes in IMT (p<0.001), RI (p<0.001) and PI (p<0.001). IMT correlated with RI (r=0.25; p<0.001), PI (r=0.26; p<0.001), and RA/CR ratio (r=0.43; p<0.001). CONCLUSIONS: Carotid IMT and intra-renal resistances are elevated at an early stage in MetS and are associated with a dysregulated production of fat-derived hormones and cytokines. PMID- 19815302 TI - Association of PTPN22 C1858T and CTLA-4 A49G polymorphisms with Type 1 Diabetes in Croatians. AB - In this case-control study the association between the PTPN22 1858T and CTLA-4 49G gene variants and T1D in Croatian population was examined. We found that distribution of PTPN22 C1858T and CTLA-4 A49G genotypes between T1D patient (n=102) and control (n=193) groups differ significantly (p<0.0001 and p=0.012, respectively). Moreover, although the risk alleles of both SNPs are distributed more frequently in patients, the significant difference is observed only for PTPN22 1858T allele (p<0.0001). This is therefore the first evidence that analyzed gene variants contribute to T1D pathogenesis in Croatian population. PMID- 19815303 TI - Ultrastructural changes in the glomerular filtration barrier and occurrence of proteinuria in Chinese patients with type 2 diabetic nephropathy. AB - AIM: Diabetic nephropathy (DN) is one of the most important causes of end stage renal disease in the world. Its hallmark is proteinuria. Therefore, we set out to clarify the structural changes that occur in the glomerular filtration barrier in Chinese patients with true type 2 diabetic nephropathy, and to examine the relationship between these structural changes and proteinuria. METHODS: 42 Chinese patients with true T2DN were divided into three groups according to urinary protein excretion. Glomerular volume, endothelial cell density, endothelial cell number, glomerular basement membrane (GBM) width, podocyte density, podocyte number and foot process width were evaluated using light and electron microscopic morphometry. RESULT: Glomerular volume and endothelial cell number were increased in diabetic patients, but there was no difference between patients with respect to the degree of proteinuria. As proteinuria progressed, endothelial cell density remained unchanged, while the glomerular basement membrane (GBM) and podocyte foot process width increased, podocyte density and number decreased. CONCLUSIONS: Podocyte and GBM change more obviously during the development of proteinuria. Besides, proteinuria was inversely related to podocyte density, and directly related to GBM and glomerular volume. PMID- 19815304 TI - The epidermal growth factor receptor ligand amphiregulin is a negative regulator of hepatic acute-phase gene expression. AB - BACKGROUND/AIMS: The modulation of the hepatic acute-phase reaction (APR) that occurs during inflammation and liver regeneration is important for allowing normal hepatocellular proliferation and the restoration of homeostasis. Activation of acute-phase protein (APP) gene expression by interleukin-6 (IL-6) type cytokines is thought to be counteracted by growth factors released during hepatic inflammation and regeneration. The epidermal growth factor receptor (EGFR) ligand amphiregulin (AR) is readily induced by inflammatory signals and plays a nonredundant protective role during liver injury. In this paper, we investigated the role of AR as a modulator of liver APP gene expression. METHODS: Expression of APP genes was measured in the livers of AR(+/+) and AR(-/-)mice during inflammation and regeneration and in cultured liver cells treated with AR and oncostatin M (OSM). Crosstalk between AR and OSM signalling was studied. RESULTS: APP genes were overexpressed in the livers of AR(-/-) mice during inflammation and hepatocellular regeneration. In cultured AR-null hepatocytes and human hepatocellular carcinoma (HCC) cells after AR knockdown, APP gene expression is enhanced. AR counteracts OSM-triggered signal transducer and activator of transcription 3 signalling in hepatocytes and attenuates APP gene transcription. CONCLUSIONS: Our data support the relevance of EGFR-mediated signalling in the modulation of cytokine-activated pathways. We have identified AR as a key regulator of hepatic APP gene expression during inflammation and liver regeneration. PMID- 19815305 TI - Porphobilinogen deaminase over-expression in hepatocytes, but not in erythrocytes, prevents accumulation of toxic porphyrin precursors in a mouse model of acute intermittent porphyria. AB - BACKGROUND & AIMS: Acute intermittent porphyria (AIP) is characterized by hepatic porphobilinogen deaminase (PBGD) deficiency resulting in a marked overproduction of presumably toxic porphyrin precursors. Our study aimed to assess the protective effects of bone marrow transplantation or PBGD gene transfer into the liver against phenotypic manifestations of acute porphyria attack induced in an AIP murine model. METHODS: Lethally irradiated AIP mice were intravenously injected with 5x10(6) nucleated bone marrow cells from wild type or AIP donor mice. To achieve liver gene transfer, AIP mice received via hydrodynamic injection plasmids expressing human PBGD or luciferase, driven by a liver specific promoter. RESULTS: Erythrocyte PBGD activity increased 2.4-fold in AIP mice receiving bone marrow cells from normal animals. Nevertheless, phenobarbital administration in these mice reproduced key features of acute attacks, such as massively increased urinary porphyrin precursor excretion and decreased motor coordination. Hepatic PBGD activity increased 2.2-fold after hydrodynamic injection of therapeutic plasmid. Mice injected with the luciferase control plasmid showed a high excretion of porphyrin precursors after phenobarbital administration whereas just a small increase was observed in AIP mice injected with the PBGD plasmid. Furthermore, motor disturbance was almost completely abolished in AIP mice treated with the therapeutic plasmid. CONCLUSIONS: PBGD deficiency in erythroid tissue is not associated with phenotypic manifestations of acute porphyria. In contrast, PBGD over-expression in hepatocytes, albeit in a low proportion, reduced precursor accumulation, which is the hallmark of acute porphyric attacks. Liver-directed gene therapy might offer an alternative to liver transplantation applicable in patients with severe and recurrent manifestations. PMID- 19815306 TI - Magnetic resonance imaging and spectroscopy for quantification of hepatic steatosis: urgent need for standardization! PMID- 19815307 TI - Personalized medicine: factors influencing reimbursement. AB - OBJECTIVES: Personalized medicine (PM) has attracted tremendous interest, but yielded few marketed products. We examined factors influencing the reimbursement of existing PM technologies. METHODS: We conducted six case studies of the following paired genetic tests and treatments: HER2/neu with trastuzumab (Herceptin); hepatitis C genotyping with ribavirin/pegylated interferon; Oncotype DX with chemotherapy; UGT1A1 with irinotecan (Camptosar); VKORC1/CYP2C9 with warfarin; BRCA1/2 with prophylactic surgical measures; and Oncotype DX with chemotherapy. We developed a framework for categorizing PM technology, and assessed factors influencing reimbursement, including quality of evidence, type of regulatory oversight, presence of clinical guidelines, and cost-effectiveness. RESULTS: PM is not a monolithic concept, but rather encompasses different types of technology. The strength of evidence available for existing PM technology varies widely and, along with endorsement of clinical guidelines, appears to be the strongest predictor of reimbursement. In the absence of reimbursement, direct to-consumer marketing has continued for some PM technology. The type of regulatory oversight and the results of cost-effectiveness analysis do not appear to be associated with reimbursement to date. CONCLUSIONS: To date, the promise and hype of PM has outpaced its evidentiary support. In order to achieve favorable coverage and reimbursement and to support premium prices for PM, manufacturers will need to bring better clinical evidence to the marketplace and better establish the value of their products. PMID- 19815308 TI - Different gene regulation strategies revealed by analysis of binding motifs. AB - Coordinated regulation of gene expression relies on transcription factors (TFs) binding to specific DNA sites. Our large-scale information-theoretical analysis of > 950 TF-binding motifs demonstrates that prokaryotes and eukaryotes use strikingly different strategies to target TFs to specific genome locations. Although bacterial TFs can recognize a specific DNA site in the genomic background, eukaryotic TFs exhibit widespread, nonfunctional binding and require clustering of sites to achieve specificity. We find support for this mechanism in a range of experimental studies and in our evolutionary analysis of DNA-binding domains. Our systematic characterization of binding motifs provides a quantitative assessment of the differences in transcription regulation in prokaryotes and eukaryotes. PMID- 19815309 TI - Treatment of subcutaneous phaeohyphomycosis and prospective follow-up of 17 kidney transplant recipients. AB - BACKGROUND: Subcutaneous phaeohyphomycosis in solid organ recipients may have an adverse outcome. OBJECTIVE: We sought to describe the disease course, treatment, and outcome of allograft function in kidney transplant recipients with phaeohyphomycosis. METHODS: Seventeen patients were followed for a mean period of 25.4 months to analyze the clinical response to treatment. RESULTS: There was no treatment failure or relapsing disease among 12 patients who completed treatment. Two patients were still in treatment with disease remission. One patient discontinued the study during treatment with partial remission, one died after finishing treatment with disease remission, and one was dropped from the study because contact was lost. Immunosuppressive regimens were not changed. Two of 17 patients had a significant reduction in allograft function. LIMITATIONS: The follow-up time was short and the number of patients was small. CONCLUSIONS: The outcome of phaeohyphomycosis in kidney transplant recipients was favorable with minimal impact on renal allograft function. PMID- 19815310 TI - Occupational exposures in emergency medical service providers and knowledge of and compliance with universal precautions. AB - BACKGROUND: Little is known about compliance with universal precautions (CUP) or occupational exposures to blood and body fluids among Emergency Medical Services (EMS) providers. The objective of this study was to obtain estimates of CUP and knowledge of universal precautions (KUP), occupational exposures, and needle and lancet sticks in the prehospital environment. METHODS: A convenience sample of workers (n=311, 51% response) from 17 agencies in Virginia that provided emergency ground transportation (volunteer, commercial, government rescue squads, and fire departments) completed a questionnaire on certification and training, KUP, CUP, exposures and needlesticks, risk perceptions, and demographic variables. RESULTS: Nearly all EMS providers reported exposures and were concerned about risk of HIV and hepatitis. Providers reported inconsistent CUP when treating patients or using needles, including failure to wear gloves (17%) and to appropriately dispose of contaminated materials (79%), including needles (87%), at all times. Certification type (advanced and basic) was related to both KUP and CUP. Of those respondents reporting current sharps use, 40% recapped needles. A lancet stick was reported by 1.4% (n=5), and 4.5% reported a needlestick (n=14). CONCLUSION: EMS providers working in the prehospital environment experience significant exposures but are not consistently using universal precautions. PMID- 19815311 TI - [Acute hepatitis due to hepatitis C virus infection in the adult population]. AB - INTRODUCTION: Knowledge of acute hepatitis C infection (AHC) is important to design effective prevention and treatment strategies. The aim of this study was to determine the clinical and epidemiological features and outcomes of patients diagnosed with AHC. METHODS: A retrospective clinical-epidemiological study was carried out in adult patients diagnosed with AHC between 2000 and 2008 in our hospital. RESULTS: Sixteen patients were diagnosed. The most probable source of transmission was intravenous drug abuse in four patients, a percutaneous injury in three healthcare workers, a medical procedure in three patients and sexual contact in two patients. The source of infection was unknown in four patients. Diagnosis was based on acute clinical symptoms in eight patients, follow-up of healthcare injury in three patients and was fortuitous in five patients. Nine patients showed severe symptoms, while seven patients were asymptomatic or had mild symptoms. The four patients who received treatment were cured. Of the 12 patients that did not receive treatment, the disease became chronic in 58.3%. Progression to chronic disease was more frequent in patients without jaundice than in those with jaundice (85.7% vs 16.7%). CONCLUSIONS: Because medical procedures are still a risk factor for hepatitis C infection, active surveillance is required as well as compliance with prevention measures against parenteral transmission. The percentage of patients with jaundice who progress to chronic disease is low and consequently the start of treatment in these patients could be delayed according to their clinical course. PMID- 19815312 TI - [Pain and unexpressed symptoms: the other dementia]. AB - Patients with advanced dementia are biologically, socially and personally highly vulnerable. The care of these patients is a challenge in terms of both the quantity of care required and qualitative aspects (the need for specific and adapted approaches). The advanced phases of dementia are characterized by severe speech impairment, loss of mobility, and feeding and nutritional alterations (in patients with severe cognitive and functional impairment). Problems of recognition and verbal expression of sensations hampers the diagnostic and therapeutic approach. This article briefly reviews the clinical characteristics of the symptoms and syndromes prevalent in these patients (pain, neuropsychiatric symptoms, delirium, epilepsy) and emphasizes the general principles for prevention and therapeutic approaches. PMID- 19815313 TI - [INCA study: cardiac failure or vascular risk?]. PMID- 19815314 TI - Synthesis and characterisation of thiosemicarbazonato molybdenum(VI) complexes and their in vitro antitumor activity. AB - New dioxomolybdenum(VI) complexes were obtained by the reaction of [MoO2(acac)2] with thiosemicarbazone ligands derived from 3-thiosemicarbazide and 4 (diethylamino)salicylaldehyde (H2L1), 2-hydroxy-3-methoxybenzaldehyde (H2L2) or 2 hydroxy-1-naphthaldehyde (H2L3). In all complexes thiosemicarbazonato ligands are coordinated to molybdenum as tridentate ONS-donors. Octahedral coordination of each molybdenum atom is completed by methanol molecule (in 1a-3a) or by oxygen atom of Mo=O unit from the neighbouring molecule (in 1-3). All complexes were characterized by means of chemical analyses, IR spectroscopy, TG and NMR measurements. The molecular structures of the ligand H2L2 and complex [MoO2L2(CH3OH)].CH3OH (2a) have been determined by single crystal X-ray crystallography. The characterisation of thiosemicarbazonato molybdenum(VI) complexes (1-4) as well as of the 4-phenylthisemicarbazonato molybdenum(VI) complexes (5-8) in aqueous medium revealed that upon dissolving complexes in water, most likely to some extent dissociation took place, although experimental data didn't allow exact quantification of dissociation. The antiproliferative effects of studied molybdenum(VI) complexes (1-8) on the human cell lines were identical to the activity of their corresponding ligands. PMID- 19815315 TI - 2-Azetidinone derivatives: design, synthesis, in vitro anti-microbial, cytotoxic activities and DNA cleavage study. AB - A novel series of 3-chloro-4-[4-(2-oxo-2H-chromen-4-ylmethoxy)-phenyl]-1-phenyl azetidin-2-one derivatives (5a-j) have been synthesized from 4 aryloxymethylcoumarins (1a-e) and 4-aryliminomethyl-phenols (3a-b). The title compounds were screened for their in vitro anti-bacterial and anti-fungal activities. Results revealed that, compounds (5c), (5f), (5h) and (5j) showed excellent anti-microbial activity against a panel of microorganisms. Brine shrimp bioassay was also carried out to study their in vitro cytotoxic properties among which (5h) and (5j) displayed potent cytotoxic activity against Artemia salina. The DNA cleavage activity of some compounds was studied by agarose gel electrophoresis method. All synthesized compounds were characterized using IR, (1)H NMR, (13)C NMR, MS and elemental analysis. PMID- 19815316 TI - Novel hybrids from lamellarin D and combretastatin A 4 as cytotoxic agents. AB - A new series of hybrids of lamellarin D and combretastatin A 4, 1,2-diphenyl-5,6 dihydropyrrolo [2,1-a] isoquinolines, were designed as cytotoxic agents based on principles of combination in medicinal chemistry and taking the parent compounds' different anti-proliferative mechanisms into consideration. Twenty-two novel hybrids were synthesized through a convenient route, with a key step of core pyrrole formation and evaluated for their anti-proliferative activities in vitro against K-562, A-549, SMMC-7721, SGC-7901 and HCT-116 cancer cell lines. The results showed that some hybrids had good anti-proliferative activities in low IC50 ranges. PMID- 19815317 TI - [Fever and abdominal pain in a 56-year-old woman]. PMID- 19815318 TI - The role of axial torque in disc herniation. AB - BACKGROUND: Epidemiological studies have found associations between lifting, lifting and twisting and twisting alone with increased incidence of disc herniation. This study investigated the role of repeated dynamic axial torque/twist combined with repeated flexion on the disc herniation mechanism. METHODS: Porcine cervical spines were tested in one of the following four testing protocols: flexion-extension only; axial torque/twist only; flexion-extension followed by axial torque/twist; or axial torque/twist followed by flexion extension. Plane film radiographs and computed tomography with contrast in the nucleus were obtained at regular intervals during and following the mechanical testing process together with final dissection to determine the disc injury patterns. FINDINGS: Axial torque/twist in combination with repetitive flexion extension motion, regardless of order, encouraged radial delamination within the annulus (67.5% of specimens). Alternatively, repetitive flexion motion alone encouraged posterior or posterolateral nucleus tracking through the annulus. Axial torque/twist alone was unable to initiate a disc herniation. Both X-ray images with contrast and computed tomography were not good at detecting radial delamination observed during dissection. INTERPRETATION: The clinical relevance is that twisting may cause more radial delamination while repeated flexion causes more posterior tracking of the nucleus giving guidance for both prevention and rehabilitation decisions. In addition, X-ray images with contrast are not effective at detecting the radial delamination which was exacerbated by combined loading in flexion extension and axial torque/twist. PMID- 19815319 TI - Response and potential of agroforestry crops under global change. AB - The use of agroforestry crops is a promising tool for reducing atmospheric carbon dioxide concentration through fossil fuel substitution. In particular, plantations characterised by high yields such as short rotation forestry (SRF) are becoming popular worldwide for biomass production and their role acknowledged in the Kyoto Protocol. While their contribution to climate change mitigation is being investigated, the impact of climate change itself on growth and productivity of these plantations needs particular attention, since their management might need to be modified accordingly. Besides the benefits deriving from the establishment of millions of hectares of these plantations, there is a risk of increased release into the atmosphere of volatile organic compounds (VOC) emitted in large amounts by most of the species commonly used. These hydrocarbons are known to play a crucial role in tropospheric ozone formation. This might represent a negative feedback, especially in regions already characterized by elevated ozone level. PMID- 19815320 TI - Environmental monitoring of Domingo Rubio stream (Huelva Estuary, SW Spain) by combining conventional biomarkers and proteomic analysis in Carcinus maenas. AB - Element load, conventional biomarkers and altered protein expression profiles were studied in Carcinus maenas crabs, to assess contamination of "Domingo Rubio" stream, an aquatic ecosystem that receives pyritic metals, industrial contaminants, and pesticides. Lower antioxidative activities - glucose-6 phosphate and 6-phosphogluconate dehydrogenases, catalase - were found in parallel to higher levels of damaged biomolecules - malondialdehyde, oxidized glutathione -, due to oxidative lesions promoted by contaminants, as the increased levels of essential - Zn, Cu, Co - and nonessential - Cr, Ni, Cd - elements. Utility of Proteomics to assess environmental quality was confirmed, especially after considering the six proteins identified by de novo sequencing through capLC-muESI-ITMS/MS and homology search on databases. They include tripartite motif-containing protein 11 and ATF7 transcription factor (upregulated), plus CBR-NHR-218 nuclear hormone receptor, two components of the ABC transporters and aldehyde dehydrogenase (downregulated). These proteins could be used as novel potential biomarkers of the deleterious effects of pollutants present in the area. PMID- 19815321 TI - Pd-nanoparticles cause increased toxicity to kiwifruit pollen compared to soluble Pd(II). AB - In the present study, endpoints including in vitro pollen performance (i.e., germination and tube growth) and lethality were used as assessments of nanotoxicity. Pollen was treated with 5-10 nm-sized Pd particles, similar to those released into the environment by catalytic car exhaust converters. Results showed Pd-nanoparticles altered kiwifruit pollen morphology and entered the grains more rapidly and to a greater extent than soluble Pd(II). At particulate Pd concentrations well below those of soluble Pd(II), pollen grains experienced rapid losses in endogenous calcium and pollen plasma membrane damage was induced. This resulted in severe inhibition and subsequent cessation of pollen tube emergence and elongation at particulate Pd concentrations as low as 0.4 mg L(-1). Particulate Pd emissions related to automobile traffic have been increasing and are accumulating in the environment. This could seriously jeopardize in vivo pollen function, with impacts at an ecosystem level. PMID- 19815322 TI - Are private physicians more likely to veto generic substitution of prescribed pharmaceuticals? AB - Physicians' decisions whether or not to veto generic substitution were analyzed using a sample of 350,000 pharmaceutical prescriptions from the county of Vasterbotten, Sweden. Although generic substitution reforms have been introduced in many European countries and American states, this is to my knowledge the first study on this topic. The topic is important since physicians' decisions regarding generic substitution not only directly affect patients' and insurers' costs for pharmaceuticals, but also indirectly since more bans against substitution reduces price-competition between pharmaceutical firms. The primary purpose was to test if physicians working at private practices were more likely to oppose substitution than county-employed physicians working on salary. It was found that private physicians were 50-80% more likely to veto substitution. Also, the probability of a veto was found to increase as patients' copayments decreased. This might indicate moral hazard in insurance, though other explanations are plausible. PMID- 19815323 TI - Visual expertise does not predict the composite effect across species: a comparison between spider (Ateles geoffroyi) and rhesus (Macaca mulatta) monkeys. AB - Humans are subject to the composite illusion: two identical top halves of a face are perceived as "different" when they are presented with different bottom halves. This observation suggests that when building a mental representation of a face, the underlying system perceives the whole face, and has difficulty decomposing facial features. We adapted a behavioural task that measures the composite illusion to examine the perception of faces in two nonhuman species. Specifically we had spider (Ateles geoffroyi) and rhesus monkeys (Macaca mulatta) perform a two-forced choice, match-to-sample task where only the top half of sample was relevant to the task. The results of Experiment 1 show that spider monkeys (N=2) process the faces of familiar species (conspecifics and humans, but not chimpanzees, sheep, or sticks), holistically. The second experiment tested rhesus monkeys (N=7) with the faces of humans, chimpanzees, gorillas, sheep, and sticks. Contrary to prediction, there was no evidence of a composite effect in the human (or familiar primate) condition. Instead, we present evidence of a composite illusion in the chimpanzee condition (an unfamiliar primate). Together, these experiments show that visual expertise does not predict the composite effect across the primate order. PMID- 19815324 TI - Pulmonary inflammatory myofibroblastic tumour: a confusing diagnosis. PMID- 19815325 TI - Lung resections in patients with only one lung. PMID- 19815326 TI - [Hypersensitivity pneumonitis after exposure to Candida spp]. AB - Hypersensitivity pneumonitis (HP) is a lung disease caused by heavy and recurrent inhalation of antigens. We describe the case of a patient with HP caused by domestic exposure to Candida spp. The diagnosis was made by taking into consideration the, clinical presentation, exposure history, radiological findings, bronchoalveolar lavage, lung function and the immuno-allergy study. The diagnosis was definitively confirmed by performing a specific bronchial provocation test. It has been shown that there is cross-reactivity between different Candida species, and despite making the diagnosis in this case with Candida albicans, we were unable to define exactly which species was responsible for the HP. PMID- 19815327 TI - [Sleep medicine and transport workers. Medico-social aspects with special reference to sleep apnoea syndrome]. PMID- 19815328 TI - [Assessment of pulmonary function prior to lung resection]. PMID- 19815329 TI - The sensitivity and specificity of a new formula to distinguish endometrioid type endometrial carcinoma from ovarian endometrial carcinoma. AB - OBJECTIVES: Endometrioid type adenocarcinoma sometimes occupies both endometrium and ovary and in some cases the origin cannot be determined. STUDY DESIGN: In this study, we established a formula to distinguish ovarian endometrioid cancer (EOC) from endometrioid type endometrial cancer (EEC), based on our previous report of cyclin and KI67 expression pattern by immunohistochemistry of 36 EECc and 37 OECc by the logistic regression. We calculated the diagnostic accuracy using 92 test samples retrospectively and finally could diagnose the origin of 16 cases in whom endometrioid type adenocarcinoma arose in both ovary and endometrium and could be determined by Scully's criteria, and 15 cases in whom endometrioid type adenocarcinoma arose in both ovary and endometrium and Scully's criteria were not useful retrospectively. RESULTS: The estimated formula is as follows: Logit(Prob(EOC))=-1.1437-0.0853 CNA+0.0423 CNB+0.173 CND1+0.0129 CNE+0.0224 CNF+0.0508 KI67, where Prob(EOC) is the probability that a clinical sample is EOC. If Prob(EOC) is larger than 0.5, the diagnosis is ovarian cancer; if less than 0.5 it is endometrial cancer. Finally, using the formula, 37 of 48 EECs (77.1%) and 33 of 44 EOCs (75.0%) were correctly classified, with an accuracy of 76.1% (p<0.0001), retrospectively. In 12 of the 16 cases (75%) who could be determined by Scully's criteria, the origin determined by Scully's criteria was concordant with the origin determined by the formula retrospectively. In the other 15 cases, 12 cases were judged as ovary/ovary, 2 cases were judged as uterus/uterus and 1 case was judged as uterus/ovary. CONCLUSION: The formula we established was thought to be useful to distinguish the origin of the cases in whom endometrioid type adenocarcinoma arises in both ovary and endometrium. PMID- 19815330 TI - One mole may hide another: The case of an ovarian hydatidiform mole coexistent with intrauterine hydatidiform mole. PMID- 19815331 TI - A double nucleotide insertion-induced frame-shift mutation of the androgen receptor gene in a familial complete androgen insensitivity syndrome. AB - OBJECTIVE: A wide spectrum of androgen receptor (AR) gene mutations has been reported in complete androgen insensitivity syndrome (CAIS). The molecular basis of androgen resistance was investigated in a female with familial CAIS. STUDY DESIGN: AR gene and protein were investigated by PCR and direct sequencing and Western immunoblotting, respectively. RESULTS: Sequencing analysis of DNA of the patient identified a double nucleotide insertion in exon 4 that results in the frame-shift leading to premature terminal signal in the beginning of exon 6. This mutation predicted the synthesis of a truncated AR that lacks the entire ligand binding molecules. Immunoblotting analysis of the gonad removed from the patient detected the mutated AR protein of 94 kDa. Positive control revealed the normal apparent molecular mass of 110 kDa. DNA sequencing of her mother demonstrated the presence of both canonical and mutated sequences in the exon 4 through 8. CONCLUSION: These findings suggested that the previously undescribed insertion mutation in the AR gene is the cause of CAIS in this family. PMID- 19815332 TI - DNA-templated photo-induced silver nanowires: fabrication and use in detection of relative humidity. AB - A very simple and novel approach of fabricating Ag-DNA network is herein reported. The Ag-DNA network can be formed from reduction of silver ion absorbed on DNA template by sunlight. Mesh size of the Ag-DNA network and the diameter of the Ag-DNA nanowire can be controlled by adjusting the concentration of DNA and irradiation time, respectively. Furthermore, the Ag-DNA network placed onto comb like gold electrodes can be utilized as a sensor for humidity, which presents a good response to the detection of relative humidity (RH). PMID- 19815333 TI - Denitrification using a monopolar electrocoagulation/flotation (ECF) process. AB - Nitrate levels are limited due to health concerns in potable water. Nitrate is a common contaminant in water supplies, and especially prevalent in surface water supplies and shallow wells. Nitrate is a stable and highly soluble ion with low potential for precipitation or adsorption. These properties make it difficult to remove using conventional water treatment methods. A laboratory batch electrocoagulation/flotation (ECF) reactor was designed to investigate the effects of different parameters such as electrolysis time, electrolyte pH, initial nitrate concentration, and current rate on the nitrate removal efficiency. The optimum nitrate removal was observed at a pH range of between 9 and 11. It appeared that the nitrate removal rate was 93% when the initial nitrate concentration and electrolysis time respectively were 100 mg L(-1)-NO(3)( ) and 40 min. The results showed a linear relationship between the electrolysis time for total nitrate removal and the initial nitrate concentration. It is concluded that the electrocoagulation technology for denitrification can be an effective preliminary process when the ammonia byproduct must be effectively removed by the treatment facilities. PMID- 19815334 TI - Endovascular treatment for moyamoya disease in a Caucasian twin with angioplasty and Wingspan stent. AB - The pathogenesis and genetics of moyamoya disease (MMD) remain a mystery. Here we report a case of female Caucasian adult twins with similar presentations of bilateral MMD that were treated with different surgical modalities. One twin was treated with a bypass and remains event free (defined as either transient ischemic event or stroke) at her 4-year follow-up. The second twin underwent angioplasty to treat a left middle cerebral artery stenosis as well as the placement of a Wingspan stent to treat a right supraclinoid ICA stenosis on separate occasions. The left middle cerebral artery angioplasty thrombosed due to the discontinuation of clopidogrel resulting in recurrent symptoms, principally aphasia, and the Wingspan stent underwent angioplasty for in-stent stenosis. Despite this, the stent has been patent and the patient has remained event free at her 2-year follow-up, post-placement. The relatively new Wingspan stent has been used in the past for intracerebral atherosclerotic lesions, however, this is the first report of it being used in a case of MMD. An overview of the MMD literature in twins and endovascular therapy is given. PMID- 19815335 TI - The activity of a designer tissue inhibitor of metalloproteinases (TIMP)-1 against native membrane type 1 matrix metalloproteinase (MT1-MMP) in a cell-based environment. AB - The surface-anchored membrane type 1 matrix metalloproteinase (MT1-MMP) degrades a wide range of extracellular matrix components that includes collagens, laminins, fibronectin and the structural proteoglycan aggrecan. The enzyme modulates cell motility and plays an important role in tumour invasion and proliferation. We have previously designed a variant of tissue inhibitor of metalloproteinase (TIMP)-1 bearing a triple mutation (V4A+P6V+T98L, or N-TIMP 1(mt1)) that forms tight binary complex with the soluble catalytic domain of MT1 MMP [M.H. Lee, M. Rapti, G. Murphy, J. Biol. Chem. 278 (2003) 40224-40230]. Here, we report our latest findings on the cellular potency of this mutant against native MT1-MMP in cell-based environment. We show that N-TIMP-1(mt1) is a highly potent inhibitor against the ectodomain form of MT1-MMP (K(i) 9.53nM) with potential for further development as a therapeutic agent. The mutant is devoid of pro-MMP-2-activating capability but is highly effective in blocking MT1-MMP mediated FITC-labelled collagen and gelatin film degradation in HTC75 fibrosarcoma and MCF7 breast cancer models. Most encouragingly, N-TIMP-1(mt1) is also effective against CD44 shedding in HTC75 cells and able to prevent tubule formation in human umbilical vascular endothelial cells (HUVEC) in a 3D fibrin gel model. We are interested in the development of the TIMPs as therapeutic agents against MT1-MMP related disorders such as cancers. Our findings here indicate the potential for the design of selective TIMPs with refined specificity and possibility for future therapeutic application. PMID- 19815336 TI - MSX1 induces the Wnt pathway antagonist genes DKK1, DKK2, DKK3, and SFRP1 in neuroblastoma cells, but does not block Wnt3 and Wnt5A signalling to DVL3. AB - Neuroblastoma is the most common extra-cranial solid childhood cancer; it arises from neural crest-derived cells of the sympathetic nervous system. The anomalous regulation of embryonic developmental pathways like Delta-Notch and Wnt has been implicated in aberrant cell growth and differentiation in many (childhood) tumours. We have previously found regulation of Delta-Notch pathway genes by the MSX1 neural crest development gene in a neuroblastoma cell line, and significant correlations between these genes in neuroblastic tumours. However, a clear role for the Wnt pathway in neuroblastic tumours has not yet been determined. We now analyze the complete spectrum of genes regulated by inducible expression of MSX1 in the SJNB8 neuroblastoma cell line using Affymetrix expression profiling. We show that MSX1 induces the expression of four different Wnt pathway inhibitor genes: Dickkopf 1-3 (DKK1-3) and secreted frizzled-related protein 1 (SFRP1), and provide evidence that high expression of two of these genes correlates with good prognosis. We were able to demonstrate that both the canonical Wnt3 and the alternative Wnt5A ligands are highly expressed in neuroblastic tumours and cell lines, and specifically activate the DVL3 Wnt co-receptor protein in SJNB8 neuroblastoma cells. These results suggest involvement of MSX1 in Wnt signalling and demonstrate activity of the more upstream Wnt pathway in neuroblastic cells. PMID- 19815337 TI - Dissolution of a new explosive formulation containing TNT and HMX: comparison with octol. AB - GIM (Greener Insensitive Material) is a new explosive formulation made of HMX (51.5%), TNT (40.7%), and a binder, ETPE (7.8%), which is currently investigated by the Canadian Department of National Defense for a wider use by the Army. In the present study, dissolution of GIM in water was measured and compared to the dissolution of octol (HMX/TNT: 70/30). Although the presence of ETPE did not prevent completely TNT and HMX from dissolving, GIM appeared to dissolve more slowly than octol. The ETPE was shown to prevent the formulation particles from collapsing and to retard the dissolution of both TNT and HMX by limiting their exposure to water. In both octol and GIM, the dissolution rate of the particles was governed by the compound(s) that are slower to dissolve, i.e. HMX in octol, and HMX and ETPE in GIM. A model based on Fick's diffusion law allowed fitting well the dissolution data of octol but was less appropriate to fit the data of GIM likely due to a physical rearrangement of the solid upon dissolution. The present findings demonstrate that ETPE in GIM decreases the risks of explosives leakage from particles of the new formulation and should facilitate the collecting of non-exploded GIM particles in training sites. PMID- 19815338 TI - Removal of tinidazole from waters by using ozone and activated carbon in dynamic regime. AB - The main objective of the present study was to analyze the efficacy of technologies based on ozone and activated carbon in dynamic regime to remove organic micropollutants from waters, using the antibiotic tinidazole (TNZ) as a model compound. Results obtained in static regime show that the presence of activated carbon (GAC) during tinidazole ozonation: (i) increases its removal rate, (ii) reduces oxidation by-product toxicity, and (iii) reduces the concentration of dissolved organic matter. Study of the ozone/activated carbon system in dynamic regime showed that ozonation of tinidazole before the adsorption process considerably improves column performance, increasing the volume of water treated. It was observed that the efficacy of the treatment considerably increased with a shorter contact time between TNZ and O(3) streams before entering the column allowing a much higher volume of TNZ solution to be treated compared with the use of activated carbon alone, and reducing by 75% the amount of activated carbon required per unit of treated water volume. TNZ removal by the O(3)/GAC system is lower in natural waters and especially in wastewaters, than in ultrapure water. The toxicity results obtained during TNZ treatment with O(3)/GAC system showed that toxicity was directly proportional to the concentration of TNZ in the effluent, verifying that oxidation of the organic matter in the natural waters did not increase the toxicity of the system. PMID- 19815339 TI - Chemical and structural evaluation of activated carbon prepared from jute sticks for Brilliant Green dye removal from aqueous solution. AB - Activated carbons have been prepared from jute sticks by chemical activation using ZnCl(2) and physical activation using steam for the removal of Brilliant Green dye from aqueous solution. The activated carbons and charcoal prepared from jute sticks were characterized by evaluating the surface chemistry, structural features and surface morphology. The maximum BET surface area was obtained to be 2304 m(2)/g for chemical activated carbon (ACC) while it is 730 and 80 m(2)/g for steam activated carbon (ACS) and charcoal, respectively. The FT-IR spectra exhibited that the pyrolysis and steam activation of jute sticks resulted in the release of aliphatic and O-containing functional groups by thermal effect. However, the release of functional groups is the effect of chemical reaction in the ZnCl(2) activation process. A honeycomb-type carbon structure in ACC was formed as observed on SEM images. Although charcoal and ACC were prepared at 500 degrees C the ACC exhibited much lower Raman sensitivity due to the formation of condensed aromatic ring systems. Due to high surface area and high porous structure with abundance of functional groups, the ACC adsorbed dye molecules with much higher efficiency than those of ACS and charcoal. PMID- 19815340 TI - Electrochemical treatment of phenolic waters in presence of chloride with boron doped diamond (BDD) anodes: experimental study and mathematical model. AB - This work deals with an experimental and numerical study on the electrochemical treatment of waters containing phenolic compounds with boron-doped diamond (BDD) anodes. Anodic oxidation of m-cresol, as a model of phenolic compound, was investigated by galvanostatic electrolyses. The electrolyses were carried out under different experimental conditions by using an impinging-jet flow cell inserted in a hydraulic circuit in a closed loop. On the basis of the experimental results a mathematical model was implemented to simulate the effect of the chemistry of organic compounds and solution on the process, in particular the effect of chlorides on the kinetics of m-cresol oxidation. The effect of hydrodynamics of the cell on the mass transfer towards the electrode surface was also considered. The model was validated through comparison with experimental data: the results showed that the proposed model well interpreted the complex effect on removal efficiency of such operative parameters as current density, hydrodynamic of the reactor and chemistry of the solution. The model predictions were utilised to obtain quantitative information on the reaction mechanism, as well as to predict the performance of the process under different operative conditions, by calculating some relevant figures of merit. PMID- 19815341 TI - Cloud point extraction with/without chelating agent on-line coupled with inductively coupled plasma optical emission spectrometry for the determination of trace rare earth elements in biological samples. AB - The on-line incorporation of cloud point extraction (CPE) with/without 8 hydroxyquinoline (8-Ox) as chelating agent into flow injection analysis associated with inductively coupled plasma optical emission spectrometry (ICP OES) for determining trace rare earth elements (REEs) is presented and evaluated. The significant parameters affecting on-line cloud point extraction of REEs such as sample pH, flow rate, 8-Ox concentration, Triton X-114 concentration were systematically studied. Under the optimized conditions, with the consumption of 3.0 mL sample solution, the limits of detection (3 sigma) were ranged from 41.4 pg mL(-1) (Yb) to 448 pg mL(-1) (Gd) with relative standard deviations (RSDs) of 1.0% (Eu)-5.9% (Sm) for on-line CPE-ICP-OES with 8-Ox as chelating agent, and 69.0 pg mL(-1) (Sc) to 509.5 pg mL(-1) (Sm) with RSDs of 2.9% (Yb)-7.5% (Ho) for on-line CPE-ICP-OES without 8-Ox as chelating agent, respectively. The sample throughput of 17samples h(-1) was obtained for both systems. The developed methods of on-line CPE-ICP-OES were validated by the analysis of certified reference material (GBW07605, tea leaves) and real biological samples of pig liver, Auricularia auricula and mushroom. PMID- 19815342 TI - Removal of direct azo dyes and aromatic amines from aqueous solutions using two beta-cyclodextrin-based polymers. AB - Two beta-cyclodextrin (beta-CD)-based polymers were synthesized using 4,4' methylene-bis-phenyldiisocyanate (MDI) or hexamethylenediisocyanate (HMDI) as a cross linking agent in dimethylformamide and used as sorbents for the removal of azo dyes (Evans Blue and Chicago Sky Blue), as well as aromatic amines (benzidine, p-chloroaniline and alpha-naphthalamine) from aqueous solutions. The sorption experiments were carried out by using batch-wise procedure involving the determination of pH effect, sorbate concentration and contact time. Moreover, from the equation isotherms such as Langmuir and Freundlich were successfully applied to model the experimental data. From the results polymer 2 was found to be a better sorbent for both azo dyes and the aromatic amines as compared to polymer 1 as proved by Langmuir isotherm model. The proposed sorption mechanism involved several kinds of interactions: physical adsorption, hydrogen bonding and formation of an inclusion complex due to the beta-CD molecules through host-guest interactions. PMID- 19815343 TI - Adsorptive removal of cationic surfactants from aqueous solutions onto high-area activated carbon cloth monitored by in situ UV spectroscopy. AB - Activated carbon cloth (ACC) was used as adsorbent for the removal of cationic surfactants such as benzyltrimethylammonium chloride (BTMACl), benzyltriethylammonium chloride (BTEACl), benzyltributylammonium chloride (BTBACl), benzyldimethyldecylammonium chloride (BDMDACl), benzyldimethyltetradecyl ammonium chloride (BDMTDACl), benzyldimethylhexadecylammonium chloride (BDMHDACl), N-dodecylpyridinium chloride (N-DPCl) and N-cetylpyridinium chloride (CPCl) from aqueous solutions. The adsorption efficiency of the ACC was evaluated for cationic surfactants. Adsorption process was followed by in situ UV spectroscopic technique. The kinetic data, so obtained, were treated according to the pseudo first-order, the pseudo second-order, the Elovich and the intraparticle diffusion models in order to understand the adsorption mechanism of cationic surfactants onto the ACC. The best fit was found with the pseudo second-order model. The experimental isotherm data were obtained at 30 degrees C and analyzed by the Freundlich and the Langmuir models. The parameters of isotherm equations were determined. The Freundlich model was found to represent the experimental data better than the Langmuir model. The observed adsorption behaviors are discussed in terms of the pH of the solution, the nature of cationic surfactants (e.g. functional groups, size, and hydrophobicity) and the nature of the ACC (e.g. surface charge, pore size). A fair linear correlation was found between some adsorption parameters and apparent molar volumes at infinite dilution for benzyltrialkylammonium chlorides. PMID- 19815344 TI - Comparison of photodegradative efficiencies and mechanisms of Victoria Blue R assisted by Nafion-coated and fluorinated TiO2 photocatalysts. AB - The purposes of this research were to study the effects of two modified photocatalysts, Nafion-coated TiO(2) and fluorinated TiO(2), and photocatalytic degradation of Victoria Blue R in aqueous solution. Photocatalytic degradation of Victoria Blue R was accelerated by the modified photocatalysts. Bulk and surface characterizations of the resulting powders were carried out. Attachment of the anions to the TiO(2) surface using the Nafion-coated-TiO(2) possibly results in increased adsorption of the cationic dye, and the degradation rate is larger for the cationic dye. It was found that Victoria Blue R on the two illuminated TiO(2) surfaces underwent very different changes. To obtain a better understanding on the mechanistic details of this modified-TiO(2)-assisted photodegradation of the Victoria Blue R dye with UV irradiation, a large number of intermediates of the process were separated, identified, and characterized by a high-performance liquid chromatography-mass spectrometry technique. Several probable photodegradation pathways were proposed and discussed. PMID- 19815345 TI - Stabilization/solidification of petroleum drill cuttings: leaching studies. AB - This work explores the effectiveness of Portland cement (CEM I), with the addition of high carbon fly ash (HCFA), as a novel binder, for the improvement of leachability-related properties of stabilized/solidified (s/s) petroleum drill cuttings. A factorial design experiment was adopted to investigate the effects of waste-to-binder ratio, HCFA addition, and curing time on leachate pH, acid neutralization capacity (ANC), and metal, chloride and hydrocarbon leaching. The leachate pH and ANC of all products suggested successful formation of a calcium silicate-hydrate-based matrix with good resistance to acid attack, and little detrimental effect from drill cuttings addition. Leaching of amphoteric metals was significantly affected by pH, which was a function of other studied factors. All studied factors also affected leaching of chloride and hydrocarbons. CEM I, without HCFA addition, was more effective in immobilizing chlorides, but the overall chloride immobilization was poor in all runs. HCFA addition significantly reduced the leaching of hydrocarbons. Comparison of milligram of contaminant leached per kilogram of drill cuttings from the s/s products and untreated drill cuttings provided clear evidence of hydrocarbon and chloride immobilization. This work shows that HCFA improved the immobilization of organic contaminants and may represent an inexpensive binder for stabilization/solidification of organic wastes. PMID- 19815346 TI - A prospective study of persistence in the prediction of smoking cessation outcome: results from a randomized clinical trial. AB - Research has had mixed success in identifying pretreatment variables which can be used to guide treatment and enhance outcome. A critical first step in the process is to identify variables that reliably predict outcome. Some recent studies, largely retrospective, have found mixed evidence on the relationship between task persistence and smoking outcome measures. In the present study, we use data from a randomized clinical trial (N=241) to prospectively investigate the ability of persistence to predict outcome. Findings from multivariate analyses did not support our hypotheses: persistence did not predict outcome. We discuss these findings in relation to previous studies by focusing on theoretical and measurement issues related to the study of persistence in smoking cessation research. We conclude by recommending directions for future research, including conceptual clarification of the relationship between persistence and theoretically related constructs and investigations of variables that may moderate relationships between these constructs and cessation outcome. PMID- 19815347 TI - The mirror neuron system may play a role in the pathogenesis of mass hysteria. AB - Mass hysteria or collective hysteria usually begins when an individual shows a hysteric manifestation in front of others in the same group who later contagiously acquire the same symptoms. The underlying pathogenesis of mass hysteria is still unknown. It has been demonstrated that the mirror neuron system (MNS) provides an important neural substrate for humans' ability to imitate and there is an inhibitive component of MNS keeping us from imitating everything we see. We proposed that the inhibitive component for MNS automatic imitation may not function well in individuals of the group that results in the outbreaks of mass hysteria. We also provide evidences from emotional contagion, gender difference and treatment in mass hysteria to support this hypothesis. PMID- 19815348 TI - Acetaminophen and neural degeneration: is there a possible link? PMID- 19815349 TI - A new hypothesis of cause of syncope: trigeminocardiac reflex during extraction of teeth. AB - Transient Loss Of Consciousness (TLOC) or vasovagal syncope is well known phenomenon in dental/maxillofacial surgery. Despite considerable study of vasovagal syncope, its pathophysiology remains to be fully elucidated. After having encountered a case of trigeminocardiac reflex after extraction of maxillary first molar we observed and studied 400 extractions under local anesthesia to know the relation between trigeminocardiac reflex and syncope. We make hypothesis that trigeminocardiac reflex which is usually seen under general anesthesia when all sympathetic reflexes are blunted can also occur under local anesthesia during extractions of maxillary molars (dento-cardiac reflex) and mediate syncope. PMID- 19815350 TI - Does maternal saliva contain fetal DNA usable for prenatal diagnostics? AB - Non-invasive molecular analysis of fetal DNA is the diagnostic goal of prenatal medicine. Circulating fetal DNA can be detected in maternal plasma. Recently, it has been detected in the urine of pregnant women. We hypothesize that fetal DNA is present also in maternal saliva and that advances in stabilization and isolation of nucleic acids from saliva enable non-invasive and repeated sampling for prenatal diagnostics. The hypothesis is testable using saliva samples of pregnant women with confirmed male fetuses. Y-specific sequences should be detectable in salivary DNA. Caution must be given to the prevention of contamination. If proved in large studies, the presence of fetal DNA fragments in maternal saliva would enable a wide range of applications in prenatal medicine. PMID- 19815351 TI - Long-term memory in brain magnetite. AB - Despite theoretical and experimental efforts to model neuronal networks, the origin of cerebral cognitive functions and memory formation are still unknown. Recently, we have proposed that in addition to chemical and electrical signals, the cellular components of the neocortex (especially neurons and astrocytes) may communicate with each other through magnetic signals generated by themselves. This magnetic communication would be the ground of short-term memory. In the present paper, we propose that brain magnetite may be a component of the mechanisms, conserved during evolution, to detect and transduce magnetic fields generated inside the cerebral neocortex. Specifically, we propose a possible role for magnetite nanoparticles, distributed through neuronal and astroglial membranes, in perception, transduction and storage of information that arrives to the neocortex. PMID- 19815353 TI - Experimental infection model for Johne's disease using a lyophilised, pure culture, seedstock of Mycobacterium avium subspecies paratuberculosis. AB - Johne's disease is a severe chronic enteritis of ruminants caused by Mycobacterium avium subspecies paratuberculosis (Map). Repeatable infections of known duration are required for validation of new diagnostic tests, evaluation of pathogenesis and development of improved vaccines. In the first study of its type, a standardised experimental model for Johne's disease was developed based on a lyophilised, low passage, pure culture, seedstock of Map. Experimental inoculations of sheep with accurately enumerated doses of Map resulted in infection outcomes across multiple trials that were modulated by the interval between inoculation and examination. Compared to an inoculum consisting of an intestinal mucosal homogenate from a naturally affected sheep, clinical signs from the pure culture of Map were manifested later, but other measures of infection were similar. Immunological assays showed that most of the inoculated animals were IFN-gamma positive in the early stages of the infection. Over time, an increasing number of sheep became Map-specific antibody positive, developed typical histopathological lesions and shed Map in their faeces. The repeatability and utility of this experimental infection model will enable study of many aspects of Johne's disease. It is the first study to show that models for Johne's disease can be standardised in relevant species using traditional microbiological approaches to production and storage of seedstock. It is recommended that an international bank of master seedstock be established, containing low passage isolates that are representative of the major strains of Map, S and C. PMID- 19815352 TI - Methamphetamine use parameters do not predict neuropsychological impairment in currently abstinent dependent adults. AB - Methamphetamine (meth) abuse is increasingly of public health concern and has been associated with neurocognitive dysfunction. Some previous studies have been hampered by background differences between meth users and comparison subjects, as well as unknown HIV and hepatitis C (HCV) status, which can also affect brain functioning. We compared the neurocognitive functioning of 54 meth dependent (METH+) study participants who had been abstinent for an average of 129 days, to that of 46 demographically comparable control subjects (METH-) with similar level of education and reading ability. All participants were free of HIV and HCV infection. The METH+ group exhibited higher rates of neuropsychological impairment in most areas tested. Among meth users, neuropsychologically normal (n=32) and impaired (n=22) subjects did not differ with respect to self-reported age at first use, total years of use, route of consumption, or length of abstinence. Those with motor impairment had significantly greater meth use in the past year, but impairment in cognitive domains was unrelated to meth exposure. The apparent lack of correspondence between substance use parameters and cognitive impairment suggests the need for further study of individual differences in vulnerability to the neurotoxic effects of methamphetamine. PMID- 19815354 TI - Decreased CD47 expression during spontaneous apoptosis targets neutrophils for phagocytosis by monocyte-derived macrophages. AB - BACKGROUND: Neutrophils (PMN) are the primary leukocyte responders during acute inflammation. After migrating into the tissues, PMN undergo programmed cell death (apoptosis) and are subsequently removed via phagocytosis by resident macrophages during the resolution phase. Efficient phagocytosis of apoptotic neutrophils is necessary for successful resolution. CD47 plays a critical role in mediating the phagocytic response, although its role in the phagocytosis of apoptotic PMN is incompletely understood. AIMS: In the present study we tested the hypotheses that CD47 modulates the targeting of apoptotic PMN for phagocytosis, and that this process is altered in neonatal PMN. STUDY DESIGN: Adult and neonatal PMN were examined for their expression of CD47. To investigate CD47-mediated functions, apoptotic adult and neonatal PMN were co-cultured with monocyte-derived macrophages (MDM) and the phagocytic index was determined using a flow cytometry based assay. RESULTS: We observed lower basal surface CD47 levels on neonatal vs. adult PMN. In both groups, spontaneous apoptosis led to decreased surface and total cellular CD47 expression. Adult and neonatal MDM ingested apoptotic neonatal target PMN more avidly than apoptotic adult target PMN. Masking of surface CD47 on PMN with a monoclonal antibody enhanced MDM phagocytic activity. CONCLUSIONS: Our results suggest that age-dependent expression of CD47 on PMN may account for differences in their ingestion by macrophages and in the resolution of inflammation. PMID- 19815355 TI - Hormonal, biochemical, and hematological profiles in female camels (Camelus dromedarius) affected with reproductive disorders. AB - The aim of this study was to assess the blood profiles in female camels affected with common reproductive disorders. Estradiol-17beta (E(2)), progesterone (P(4)), thyroxin (T(4)), zinc (Zn), copper (Cu), calcium (Ca), phosphorus (P), magnesium (Mg), cholesterol, glucose, triglycerides, total protein, albumin, globulin, hematocrite, and total and differential white blood cell counts (WBC) were determined in blood of female camels affected with endometritis (n=15), vaginal adhesions (n=15), and ovarian cysts (n=15). Normal cyclic animals were used as controls (n=15). Diagnosis of reproductive disorders was based on transrectal palpation, ultrasonographic examination, and exploration of the vagina. Increased WBC counts (P=0.03) and a tendency for neutrophelia (P=0.05) were noted in female camels with vaginal adhesions. These animals were also characterized by having higher concentration of serum P(4) (P=0.0001), T(4) (P=0.001) and total protein (P=0.007), in comparison with female camels with endometritis, ovarian cysts, or controls. Animals having ovarian cysts with thin walls and homogenous hypoechogenic contents had greater serum E(2) (P=0.001) and P(4) (P=0.0001) than those having ovarian cysts with thick walls and non-homogenous echogenic contents. Animals with endometritis, vaginal adhesions, and ovarian cysts revealed lower serum Zn concentration than that of control group (P=0.003). Other blood parameters did not differ significantly compared to controls. In conclusion, this is the first report characterizing blood constituents in female camels with various reproductive disorders. These profiles may be valuable in clarifying the etio-pathogenesis of these disorders. PMID- 19815356 TI - Effects of sleep disturbance on the quality of life of Turkish menopausal women: a population-based study. AB - OBJECTIVES: The purpose of this study was to investigate sleep disturbances among menopausal women: their prevalence, risk factors for them and the quality of life of women who have them. DESIGN: A population-based sample of 887 Turkish women aged 45-59 years and living in Malatya was recruited in this cross-sectional descriptive study. The women were administered the Interview Form, which covers sociodemographic, health and lifestyle variables, as well as the Women's Health Initiative Insomnia Rating Scale, the Menopause Specific Quality of Life Questionnaire and the Beck Depression Inventory. RESULTS: The prevalence of sleep disturbance in this sample of menopausal women was 54%. Logistic regression models revealed that the risk of sleep disturbance was 2.4 times higher in the perimenopausal than in the premenopausal period, 1.7 times higher among those who received hormone therapy than among those who did not, 1.5 times higher among those with a physical disease than among those without, and 3.9 times higher among those with depression than among those without; an increase of one year in age was associated with a 5% increase in the prevalence of sleep disturbance. Average scores on the vasomotor, psychosocial, physical and sexual sub-scales of the Menopause Specific Quality of Life Questionnaire were significantly higher for women with sleep disturbance than for those without (P<0.001). CONCLUSIONS: The prevalence of sleep disturbance was found to be high among menopausal women. Initiatives aimed at reducing sleep disturbance should be added to menopausal care programmes in order to improve the quality of life of menopausal women. PMID- 19815357 TI - Characteristics of the synchronous occipital and frontopolar spike phenomenon in Panayiotopoulos syndrome. AB - PURPOSE: The synchronous appearance of an occipital and frontopolar spike (the Fp O spike) is characteristic of Panayiotopoulos syndrome (PS). This phenomenon is also seen in various other types of epilepsy, particularly those that occur in childhood. Using dipole analysis and sequential mapping, we investigated the characteristics of the Fp-O spike observed in seven patients with PS and six patients with symptomatic localization-related epilepsy in childhood (SLE). METHODS: We analyzed both one averaged spike and 20 manually selected successive individual Fp-O spikes for each patient through sequential topographical mapping with steps of 10 ms from 40 ms before to 40 ms after the negative maximum peak of each spike. For dipole analysis, a period of 40 ms before the maximum negative peak of the averaged spike in each patient was examined using equivalent current dipole localization software. RESULTS: Sequential mapping revealed that occipital negative peaks preceded frontal negative peaks in all of the PS patients, as well as in two of the six SLE patients. The four remaining SLE patients did not exhibit preceding occipital peaks. In all of the patients with PS, representative dipole locations were in the posterior area, whereas in SLE patients they were scattered over more anterior areas. The estimated sources of the Fp-O and O spikes appeared to have the same position and orientation in the two PS patients. CONCLUSION: We conclude that Fp-O spikes in PS occur as the result of a rapid spread of epileptic activity from the posterior areas to the anterior areas of the brain. Fp-O spikes in PS patients show a uniform topographical pattern and dipole location, whereas those in other patients show more heterogeneity in these features. These findings support the homogeneity of PS and thus its designation as a syndrome. PMID- 19815358 TI - Positive association between benign familial infantile convulsions and LGI4. AB - PURPOSE: LGI4 is located in 19q13.11, where the locus of benign familial infantile convulsions (BFIC) has been mapped. LGI4 belongs to a family of proteins with the epilepsy-associated repeat (EAR) domain and is associated with various epilepsies. We investigated whether LGI4 is a candidate gene for BFIC. METHODS: Fifteen patients with BFIC were examined for mutations and/or polymorphisms of LGI4 by using a direct sequencing method. RESULTS: Several frequent polymorphisms were identified. The genotype frequency distribution of c.1722G/A polymorphism was significantly different between patients with BFIC and control subjects (p<0.05). Logistic regression analysis showed that the G allele of c.1722G/A polymorphism had significant recessive effects on the increased relative risk for BFIC (p<0.05). There was no association between c.1722G/A polymorphism and benign familial neonatal convulsion, an epilepsy phenotype similar to BFIC but genetically distinguished from BFIC. DISCUSSION: The positive genotypic association between BFIC and c.1722G/A polymorphism suggests that LGI4 might contribute to the susceptibility to BFIC. PMID- 19815359 TI - Computed tomography and magnetic resonance imaging findings of solitary fibrous tumors in the pelvis: correlation with histopathological findings. AB - OBJECTIVE: We aimed to analyze the computed tomography (CT) and magnetic resonance imaging (MRI) findings of pelvic solitary fibrous tumors (SFTs) and to improve the diagnostic efficacy for such tumors. METHODS: Six cases of pelvic SFTs confirmed by histopathology were analyzed retrospectively. Of the 6 patients, 4 had undergone CT scanning, and 2 had undergone magnetic resonance imaging. All the patients had undergone unenhanced and contrast-enhanced examinations, and 2 had also undergone dynamic CT enhancement examination. Image characteristics such as shape, size, number, edge, attenuation or intensity for each lesion before and after contrast enhancement were analyzed and compared with the pathomorphology of the tumors. RESULTS: All the 6 cases showed oval or rounded and well-defined masses. Unenhanced CT images showed heterogeneous masses with patchy, necrotic foci in 3 cases and homogeneous mass in 1 case. None of the tumors showed calcification. Contrast-enhanced CT images showed marked, heterogeneous enhancement in the first and second cases. Dynamic enhancement scan demonstrated mild homogeneous enhancement in the third case and mild prolonged, delayed enhancement and washout in the fourth case. T1-weighted MR images showed heterogeneous mild hypointense lesion with linear hyperintensity in 1 case, and homogeneous isointensity in the other. T2-weighted images showed heterogeneous mixed intensity in 1 case and mostly hyperintensive lesion with hypointense foci in another case. A case showed marked heterogeneous enhancement and another showed marked homogeneous enhancement on contrast-enhanced T1-weighted images. CONCLUSION: Radiological findings of pelvic SFTs are variable and nonspecific. However, a well-defined, ovoid or rounded mass with hypointense on MR T2-weighted images and variable enhancement on CT and MR images may suggest the diagnosis of SFTs. Pelvic SFTs should be included in the differential diagnosis of regional tumors. PMID- 19815360 TI - Development of a two-dimensional liquid chromatography system for isolation of drug metabolites. AB - The separation, isolation and identification of drug metabolites from complex endogenous matrices like urine, plasma and tissue extracts are challenging tasks. Metabolites are usually first identified by mass spectrometry and tentative structures proposed from product ion spectra. In many cases mass spectrometry cannot be used to determine positional isomers and metabolites have to be fractionated in microgram amounts for analysis by NMR. To overcome the difficulties associated with separation and isolation of drug metabolites from biological matrices, a new two-dimensional liquid chromatography system has been developed. The retention times of 45 acidic, basic and neutral compounds were determined on liquid chromatographic columns with different stationary phases in order to identify two columns with highly different selectivity to be used for two-dimensional liquid chromatography. Drug metabolites of three model compounds were first generated in vitro with liver microsomes and then compared with potential metabolites formed by oxidation with hydrogen peroxide catalyzed by meso-tetra (4-sulphonatophenyl) porphine (porphine). The results showed that the porphine system could be used as a complementary system for the generation of phase I microsomal metabolites with high yield of some metabolites in a less complex matrix. The two-dimensional liquid chromatography system was used to separate and isolate microsomal and porphine generated drug metabolites in off line and on-line mode. Finally, to verify the utility of the developed system, urine samples were spiked with metabolite standards of model compounds for separation in the two-dimensional system. Excellent separations were obtained with an amide column in the first dimension and a pentafluorophenylpropyl (PFPP) column in the second dimension. The metabolites were successfully separated from each other as well as from the complex biological matrix. The results demonstrate the applicability of the system for fractionation of drug metabolites but it could also be used in many other analytical purposes, especially for basic compounds. Trace levels of metabolites were successfully separated in the on-line mode which failed in the off-line mode. PMID- 19815361 TI - A validated stability-indicating UPLC method for desloratadine and its impurities in pharmaceutical dosage forms. AB - A novel stability-indicating gradient reverse phase ultra-performance liquid chromatographic (RP-UPLC) method was developed for the determination of purity of desloratadine in presence of its impurities and forced degradation products. The method was developed using Waters Aquity BEH C18 column with mobile phase containing a gradient mixture of solvents A and B. The eluted compounds were monitored at 280nm. The run time was 8min within which desloratadine and its five impurities were well separated. Desloratadine was subjected to the stress conditions of oxidative, acid, base, hydrolytic, thermal and photolytic degradation. Desloratadine was found to degrade significantly in oxidative and thermal stress conditions and stable in acid, base, hydrolytic and photolytic degradation conditions. The degradation products were well resolved from main peak and its impurities, thus proved the stability-indicating power of the method. The developed method was validated as per ICH guidelines with respect to specificity, linearity, limit of detection, limit of quantification, accuracy, precision and robustness. This method was also suitable for the assay determination of desloratadine in pharmaceutical dosage forms. PMID- 19815362 TI - Development and validation of a RP-HPLC method for the determination of gentamicin sulfate and its related substances in a pharmaceutical cream using a short pentafluorophenyl column and a charged aerosol detector. AB - Gentamicin sulfate is a potent broad spectrum aminoglycoside antibiotic which is used as an active pharmaceutical ingredient (API) against both Gram-positive and Gram-negative bacteria. A reversed-phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated to determine the composition of gentamicin sulfate and to estimate its related substances (without any pre- or post-column derivatization) in a pharmaceutical cream. As gentamicin has a weak UV chromophore, it is not possible to detect low levels of known and unknown related substances of gentamicin using a UV detector. In this method, a Charged Aerosol Detector (CAD) was used to obtain high sensitivity that was necessary for the intended purpose of the method. This method can separate all the analogues of gentamicin including all known and unknown related substances of the API. A short (5cmx4.6mm) pentafluorophenyl HPLC column from Restek (Allure PFP) was used with an ion-pair gradient mobile phase consisting of (A) heptafluorobutyric acid:water:acetonitrile (0.025:95:5, v/v/v) and (B) trifluoroacetic acid:water:acetonitrile (1:95:5, v/v/v). PMID- 19815363 TI - Rapid identification of Propionibacterium acnes from blood cultures by fluorescence in situ hybridization. AB - A newly designed probe for rapid identification of Propionibacterium acnes by fluorescence in situ hybridization was evaluated using 111 isolates from subculture and showed 100% sensitivity and specificity. A sensitivity of 95% and a specificity of 100% were achieved with direct application on 55 blood cultures containing Gram-positive rods. PMID- 19815364 TI - A rapid, simple, and sensitive loop-mediated isothermal amplification method to detect toxigenic Vibrio cholerae in rectal swab samples. AB - Loop-mediated isothermal amplification (LAMP) method was designed for clinical diagnosis of Vibrio cholerae carrying the ctxA gene. The detection limits of the method were 5 fg of purified genomic DNA/reaction and 0.54 CFU/reaction. The method was applied to rectal swab samples from cholera patients and healthy volunteers (19 subjects each) and yielded the same results as the "gold standard" culture method, while the polymerase chain reaction-based method failed to detect V. cholerae in 8 of the positive samples. Direct application of this LAMP method without precultivation enabled the rapid detection of 5 asymptomatic carriers from rectal swabs of 21 household contacts of cholera patients. This LAMP method could be a sensitive, specific, inexpensive, and rapid detection tool for V. cholerae carrying the ctxA gene in the clinical laboratory and in the field. PMID- 19815365 TI - Patient-clinician information engagement increases treatment decision satisfaction among cancer patients through feeling of being informed. AB - OBJECTIVE: Examine how patient-clinician information engagement (PCIE) may operate through feeling informed to influence patients' treatment decision satisfaction (TDS). METHODS: Randomly drawn sample (N=2013) from Pennsylvania Cancer Registry, comprised of breast, prostate and colon cancer patients completed mail surveys in the Fall of 2006 (response rate=64%) and Fall of 2007. Of 2013 baseline respondents, 85% agreed to participate in follow-up survey (N=1703). Of those who agreed, 76% (N=1293) completed follow-up surveys. The sample was split between males and females. The majority of participants were White, over the age of 50, married, and with a high school degree. Most reported having been diagnosed with in situ and local cancer. RESULTS: PCIE was related to concurrent TDS (beta=.06) and feeling informed (beta=.15), after confounder adjustments. A mediation analysis was consistent with PCIE affecting TDS through feeling informed. Baseline PCIE predicted feeling informed (beta=.04) measured 1 year later, after adjustments for baseline feeling informed and other confounders. Feeling informed was related to concurrent TDS (beta=.35) after confounder adjustment and follow-up TDS (beta=.13) after baseline TDS and confounder adjustment. CONCLUSION: Results suggest PCIE affects TDS in part through patients' feeling informed. PRACTICE IMPLICATIONS: PCIE may be important in determining patients' level of feeling informed and TDS. PMID- 19815366 TI - VEGF modulates the effects of gonadotropins in granulosa cells. AB - Follicle selection is associated with an increase in the expression of vascular endothelial growth factor (VEGF) and its receptors in granulosa cells, however, the roles of VEGF in regulating the function of these or other non-endothelial cells in the ovary have not been explored in detail. The current study used bovine cell cultures to investigate potential roles of VEGF in the regulation of granulosa cell function during follicle development. Granulosa cells were obtained from morphologically healthy follicles 4 to 8 mm or 9 to 14 mm in diameter (corresponding to diameters before and after the establishment of dominance, respectively, during a bovine follicular wave) and exposed to a range of VEGF concentrations (1 to 100 ng/mL) encompassing concentrations found naturally in bovine dominant follicles. A concentration of VEGF of 1 ng/mL induced significant proliferation of granulosa cells from 4- to 8-mm follicles (P=0.024) and increased the proliferative response of these cells to follicle stimulating hormone (FSH; P=0.045); whereas higher doses of VEGF had no effect on proliferation (P=0.9). Treatment with VEGF induced an overall increase in mean extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation (P=0.02). In contrast, VEGF, alone or in combination with FSH, had no effect on expression of the steroidogenic enzyme, CYP11A1, by cells from 4- to 8-mm follicles (P=0.9). Granulosa cells from 9- to 14-mm follicles responded to 1 ng/mL VEGF with an increase in expression of the ovulation-associated gene, PTGS2 (P=0.003) but higher VEGF doses had no effect (P=0.9). The PTGS2 response to 1 ng/mL VEGF was similar to that induced by treatment with luteinizing hormone (LH). Interestingly, the stimulatory effects of LH on ERK1/2 phosphorylation (P=0.003) and PTGS2 expression (P<0.01) in granulosa cells from 9- to 14-mm follicles were abolished (P=0.2) by specific chemical inhibition of VEGF receptor 2 (VEGFR2). These results suggest novel and important roles of VEGF and its receptor, VEGFR2, in mediating and/or enhancing the effects of gonadotropins in granulosa cells. PMID- 19815367 TI - [Acute pancreatitis induced by olanzapine]. PMID- 19815368 TI - [Arterial catheterization: a safe procedure?]. PMID- 19815369 TI - [Therapy education for patients receiving oral anti-coagulants vitamin K antagonists]. AB - The vitamin K antagonists (VKA) remain to this day the only oral form of therapeutic anticoagulation. Approximately 1% of the French population, mainly elderly, is treated with these anticoagulants. Oral anticoagulants have significant risks of iatrogenic complications; indeed they are the leading cause of such drug-induced complications, predominantly hemorrhages. AFSSAPS (French Drug and Medical Products Agency) clinical practice recommendations, repeatedly disseminated, emphasize the education of patients receiving VKAs. Managing oral anticoagulant treatment is challenging, with a significant risk of under- or overdosing and consequently, thrombosis or hemorrhage. The therapeutic window is narrow, multiple drug-interactions are possible, and the specific dose required for a particular individual to achieve appropriate International Normalized Ratio (INR) levels is unpredictable. The literature contains few randomized controlled trials about the efficacy of education for patients treated with oral anticoagulants. These education programs are not standardized and are therefore varied and difficult to compare. Nevertheless, studies demonstrate the importance of patient education programs in reducing the risk of hemorrhage and achieving better treatment stability. The Grenoble region hospital-community network for vascular diseases (GRANTED) has developed an education program for these patients, consisting of individual sessions for the patient and/or a friend or family member (either at a health care facility or at the patient's home), telephone support and group sessions, and using educational tools and supports. There is also a link with the general practitioner who receives a report. This approach makes it possible to adapt the educational message to individual patients and their daily lives, as well as directly involving them in the management of their treatment. PMID- 19815371 TI - Predictors of specific phobia in young women: a prospective community study. AB - Potential predictors of incidence of specific phobia were investigated within the conceptual framework of the vulnerability-stress model. At two time points separated by approximately 17 months, a community sample of 1261 German women (18 25 years of age) completed a structured interview. A broad range of potential stress- and vulnerability-related predictors was recorded at initial assessment. The strongest predictors of incidence were: high levels of preexisting psychopathology, a lack of coping skills, and a negative cognitive style. Assessing individual differences in stress and vulnerability thus seem to offer additional information about etiology of specific phobia beyond traditional learning theory. Incorporating the role of these risk factors may be useful for identifying individuals who are at increased risk and improving measures of prevention. PMID- 19815370 TI - [Screening for carbon monoxide (CO) poisoning by general practitioners in home visits]. AB - OBJECTIVE: To evaluate the interest of carbon monoxide (CO) detector used by general practionners visiting patients at home. METHODS: CO detector (cost: 200 euros) was attributed to 300 general practionners visiting at least 20 patients at home per week. Alarm was triggered when ambient CO concentration exceeded 80ppm. Measurement of CO in expired breath was also possible. Activity and alarms were prospectively collected. Circumstances of intoxication were recorded. Evaluation was finally performed. The end-poind was to quantify CO-poisoning detected by the use of the device and the cost of this strategy. RESULTS: From November 2001 to November 2004, 65 scenes of intoxication with 79 victims were prospectively reported by 12 general practionners. Final evaluation revealed that 23 physicians omitted to declare alarms. Alarm incidence was of 1 for 17.527 visits; with a related cost of approximately 858 euros for 24 months. Ambient carbon monoxide concentration exceeded 200ppm in 25% of cases. Hospital admission was required for 91% of the victims. Hyperbaric oxygen therapy was performed in two cases. General practionners (n=272) considered that CO detector was useful for safety reasons (91%), they wanted to continue the experience, but did not plan to buy such device (59%). DISCUSSION: Use of CO detectors by general practionners visiting patients at home allowed to identify 65 scenes of CO intoxication. In most cases, the cause of the visit did not suggested CO poisoning. The cost of the device seems to limits its large use. CONCLUSION: CO detector is a safety tool for both general prationners and patients. Its large use has to be questioned. PMID- 19815372 TI - Are children's own interpretations of ambiguous situations based on how they perceive their mothers have interpreted ambiguous situations for them in the past? AB - The intergenerational transmission of anxious cognitions and behaviors may be a possible mechanism underlying concordance between maternal and child anxieties. By disambiguating ambiguous situations for their child in a threatening manner, anxious mothers may 'train' their children towards a general tendency to also interpret ambiguity in a threatening way. Ninety-two children and their mothers completed trait anxiety measures. Children completed two ambiguous scenario questionnaires, to measure their own interpretations of ambiguous situations (ASQ C) and to measure their expectations of their mother's interpretation and behavior in response to ambiguous situations involving them (ASQ-EM). Maternal and child anxiety were significantly correlated. Children who made threat interpretations also anticipated that their mother would disambiguate situations for them in a threatening way. The relationship between maternal anxiety and child threat cognitions was mediated by children's expectations of how their mother would disambiguate situations for them when taken together with children's trait anxiety. The present findings provide preliminary support for the suggestion that children of anxious mothers may learn to interpret and respond to ambiguous situations based on how their mothers have interpreted ambiguous situations for them in the past. PMID- 19815373 TI - Multi-sensory rooms: comparing effects of the Snoezelen and the Stimulus Preference environment on the behavior of adults with profound mental retardation. AB - The present study examined whether Snoezelen and Stimulus Preference environments have differential effects on disruptive and pro-social behaviors in adults with profound mental retardation and autism. In N=27 adults these target behaviors were recorded for a total of 20 sessions using both multi-sensory rooms. Three comparison groups were created by diagnosis and motor respective linguistic abilities. Each client was exposed to only one multi-sensory room. Results showed that Snoezelen intervention decreased disruptive behaviors only in individuals with autism, while Stimulus Preference increased pro-social behaviors only in participants with profound mental retardation with co-occurring poor motor and linguistic abilities. Furthermore, several trend analyses of the improved behaviors were conducted throughout all sessions toward short and mid term effects of the multi-sensory room applications. These findings support both the prudence of using the Snoezelen room in individuals with developmental disabilities and the importance of using a Stimulus Preference assessment in multi-sensory environments in clients with profound mental retardation. PMID- 19815374 TI - Expanding the test of counterfeit deviance: are sexual knowledge, experience and needs a factor in the sexualised challenging behaviour of adults with intellectual disability? AB - It is posited within the literature that the sexualised challenging behaviour of adults with intellectual disability may be influenced by low levels of sexual knowledge, lack of sexual experience and unmet sexual needs. In this study, individuals with sexualised challenging behaviour were identified and matched for gender, age and ability level with individuals recruited to the non-sexualised and no challenging behaviour groups. All (n=24) were interviewed using the Socio Sexual Knowledge and Attitudes Tool - Revised (SSKAAT-R) and the Sexual Knowledge, Experience and Needs Scale for Intellectual Disability (Sex-Ken-ID) to assess their sexual knowledge, experience and needs. Adaptive behaviour was measured as a covariate. In the current study, contrary to expectations in the wider literature, the sexualised challenging behaviour group showed significantly higher levels of sexual knowledge in several areas when adaptive behaviour was controlled. Their needs in relation to Dating and Intimacy were also significantly higher but no differences were found between groups in relation to sexual experience. The implications of these findings for service provision are outlined along with the considerations of directions for future research. PMID- 19815375 TI - Movement Assessment Battery for Children (M-ABC): establishing construct validity for Israeli children. AB - The Movement Assessment Battery for Children (M-ABC) is one of the most accepted tools, both in clinical practice and in research, for the diagnosis of Developmental Coordination Disorders (DCDs) in children. The present study aimed to: (1) establish the construct validity of M-ABC in Israel by comparing the motor performance of typically developed children in four age groups (ranging from 6 to 12 years) and (2) examine the impact of socio-demographic parameters on children's motor performance. Participants were 249 typical children, between 4.1 and 12.08 years old, whose motor performance was evaluated by the M-ABC. The results showed that age, gender, mother's education level, and socioeconomic status had an impact on children's motor performance. Thus, the M-ABC may serve as a suitable tool for examining the motor performance of children in Israel. This examination should also refer to socio-demographic factors in order to shed light on the contribution of environmental disadvantages to children's motor performance. PMID- 19815376 TI - Mindfulness as a potential intervention for stimulus over-selectivity in older adults. AB - Ageing is related to significant declines in cognitive functioning. This effect can have a serious impact on the physical and psychological health of older adults as well as their quality of life. One phenomenon linked to cognitive deficits, particularly attention, that has been demonstrated to emerge with ageing is over-selectivity. Over-selectivity occurs when behavior is controlled by a limited number of stimuli in the environment. Mindfulness is a construct that specifically targets attention and awareness of the present moment. The current study aimed to remediate over-selectivity in an elderly population by means of a focused attention/mindfulness induction. The results of this study indicated that the level of emergent over-selectivity in an elderly population was significantly reduced after a focused attention induction when compared to an unfocused attention induction. The findings are discussed in terms of the efficacy of mindfulness training in reducing over-selectivity. PMID- 19815377 TI - The neuropsychology of 22q11 deletion syndrome. A neuropsychiatric study of 100 individuals. AB - The primary objective of this study was to study the impact of ASD/ADHD on general intellectual ability and profile, executive functions and visuo-motor skills in children and adults with 22q11 deletion syndrome (22q11DS). A secondary aim was to study if gender, age, heart disease, ASD, ADHD or ASD in combination with ADHD had an impact on general intellectual ability and profile. One hundred consecutively referred individuals aged 1-35 years with 22q11DS were given in depth neuropsychological assessments. Mean full scale IQ was 71 with a normal distribution around this mean. Higher IQ for females than males, and a negative trend for IQ with higher age were found. Intellectual impairment, as well as visuo-motor dysfunction, was found to be related to 22q11DS per se and not to ASD/ADHD. In the area of executive function, the presence of ASD/ADHD predicted poor planning ability in the children in the study. PMID- 19815378 TI - Noise and tremor in the perception of vocal aging in males. AB - OBJECTIVE/HYPOTHESIS: To specify a set of acoustic cues for vocal aging and to establish their perceptual relevance. STUDY DESIGN: Perceptual testing. METHODS: To identify the acoustic and perceptual correlates of the aging voice, voice quality [in conjunction with speaking rate and fundamental frequency (F(0))] was systematically manipulated using resynthesis to determine its effect on perceived age. Ten young male voices were resynthesized using two levels of noise (random modulation of F(0) contour) and two levels of tremor (constant modulation of F(0) contour with a low-amplitude wave) under a speaking-rate manipulation (an increase in speaking rate that is common to older male voices). These materials were submitted to 40 naive listeners in an age-estimation task. Two sets of comparison materials were also included for evaluation: unmanipulated samples from a 150 voice database of young, middle-aged, and older voices and disordered voice samples representing natural manifestations of the voice qualities of interest. RESULTS: Speaking rate, highest degree of tremor, and highest degree of noise all shifted, in an additive manner, the mean perceived age of the young male voices by a maximum of 12 years on average; individual voices were observed being shifted by a generation. Fundamental frequency manipulations had no significant effect on perceived age. CONCLUSIONS: Voice quality (both tremor and noise) and speaking rate are all perceptually relevant cues of age in male voices. PMID- 19815379 TI - Diastolic dysfunction and its histopathological correlation in obstructive hypertrophic cardiomyopathy in children and adolescents. AB - BACKGROUND: Histopathologic hallmarks of hypertrophic cardiomyopathy (HCM) include myocyte hypertrophy and disarray as well as interstitial and endocardial fibrosis. Published correlations between echocardiographic parameters and histopathologic findings are scarce. METHODS: All patients aged <20 years (n = 45; 15 female patients; median age, 14 years) with obstructive HCM undergoing septal myectomy at the Mayo Clinic from 2003 to 2007 were identified. A retrospective review of echocardiographic data was performed, and these data were compared with the histologic findings from the myectomy specimens. RESULTS: Histopathologic analysis of myectomy specimens revealed significant myocyte hypertrophy (100%), myocyte disarray (98%), interstitial fibrosis (95%), and subendocardial fibrosis (97%). On multivariate regression analysis, there was a significant relationship between the degree of myocyte disarray and echocardiographic markers of left ventricular diastolic dysfunction. CONCLUSION: The results of this study suggest that myocyte disarray is a key factor responsible for diastolic dysfunction in pediatric patients with obstructive HCM. These findings provide novel insights into the mechanism of diastolic dysfunction in HCM that warrant further study. PMID- 19815380 TI - Continued global left ventricular remodeling is not the sole mechanism responsible for the late recurrence of ischemic mitral regurgitation after restrictive annuloplasty. AB - BACKGROUND: Recurrence of ischemic mitral regurgitation (MR) late after mitral valve annuloplasty (MVA) is generally believed to be due to continued left ventricular (LV) remodeling. The aim of this study was to determine if other mechanisms could be involved in MR recurrence. METHODS AND RESULTS: Preoperative (10 +/- 11 days), early postoperative (6 +/- 4 days), and late postoperative (1.5 +/- 0.6 years) transthoracic echocardiograms of 26 patients (68 +/- 7 years, 23 male [88%]) who underwent restrictive MVA and coronary artery bypass graft surgery were reviewed. Mitral valve geometry and MR severity were assessed using anterior leaflet and posterior leaflet angles and the anterior leaflet concavity area, defined as the area enclosed between the AL and a line connecting the tip of the leaflet and its basal insertion at the annulus. Recurrent MR (vena contracta > 3 mm) was observed in 10 patients (38.5%). Among the 10 patients with recurrent MR, 5 had significant late postoperative increase in LV end-systolic (preoperative: 66 +/- 27 mL; early postoperative: 61 +/- 11 mL; late postoperative: 89 +/- 30 mL, P = .04) and end-diastolic (preoperative: 122 +/- 39 mL; early postoperative: 108 +/- 22 mL; late postoperative: 139 +/- 39 mL, P = .04) volumes and PL angle (early postoperative: 65 +/- 12 degrees; late postoperative: 77 +/- 8 degrees, P = .04), suggesting that recurrent MR is related to continued adverse LV remodeling and ensuing worsening of leaflet tethering. However, in the remaining 5 patients with recurrent MR, there was no significant change in LV end-systolic and end-diastolic volumes, but there was a significant increase in anterior leaflet concavity area (early postoperative: 6 +/- 11 mm(2); late postoperative: 50 +/- 3 mm(2), P = .012). CONCLUSION: Although recurrent MR is often associated with continued adverse LV remodeling after restrictive MVA, this mechanism fails to explain all recurrences. In the absence of LV dilatation, recurrent MR might be explained by localized LV remodeling in the vicinity of papillary muscles resulting in increased AL tethering at the bending point. PMID- 19815381 TI - "Grey zone" patterns of unexplained endocarditis: still a challenge for clinical decision making. AB - The authors report two cases of unexplained active inflammatory endocarditis with totally different clinical presentations. The patients had undergone previous mitral repair surgery and were referred for multiple soft mobile masses on the mitral ring without clinical or laboratory signs of endocarditis. Serologic screening and blood culture results were negative, including those for specific fastidious bacteria, as well as immunologic tests to rule out "nonbacterial thrombotic endocarditis." Before new surgery, both patients were treated with long-term antibiotic and anticoagulant therapy, with no significant changes in clinical setting and echocardiographic patterns. In neither case was it possible to characterize a specific microorganism: the intraoperative findings were highly evocative of active endocarditis with a macroscopic infiltration of the mitral ring, and culture results from surgical material and valvular tissue were negative. PMID- 19815382 TI - Assessments of right ventricular volume and function using three-dimensional echocardiography in older children and adults with congenital heart disease: comparison with cardiac magnetic resonance imaging. AB - BACKGROUND: The utility of three-dimensional echocardiography (3DE) for right ventricular (RV) assessment is uncertain in older children and adults with congenital heart disease (CHD), in whom the right ventricle is often dilated and dysfunction is common. METHODS: RV assessments using 3DE were compared with manual tracing and automated border detection (ABD) with magnetic resonance imaging (MRI) as the reference method. Twenty-eight of 54 consecutive patients (52%; median age, 17 years) with CHD had adequate three-dimensional echocardiographic data sets for analysis. RESULTS: There were wide ranges of RV size (mean RV end-diastolic volume index, 143 +/- 43 mL/m(2)) and function (mean RV ejection fraction [EF], 48 +/- 10%) on MRI. End-diastolic volume was underestimated on 3DE by 20% (P < .001) and to a greater degree in larger ventricles (P < .001). There was no significant difference in EF measurements between 3DE methods and MRI except for ABD (-2.6 +/- 6, P = .03). The mean analysis time for ABD was 5 minutes, compared with 19 minutes for manual tracing (P < .0001). CONCLUSION: Approximately half the patients with CHD had adequate three-dimensional echocardiographic images. Three-dimensional echocardiography accurately estimated EF but underestimated volume, particularly when the right ventricle was dilated. ABD minimally underestimated EF but offered a significant reduction in analysis time. PMID- 19815383 TI - Comparison of left ventricular ejection fraction and left ventricular global strain as determinants of infarct size in patients with acute myocardial infarction. AB - BACKGROUND: The aim was to compare left ventricular ejection fraction (LVEF) and left ventricular (LV) global strain by speckle tracking as predictors of final infarct size. METHODS: LV global strain and LVEF by echocardiography were assessed in the acute phase and after revascularization in 39 patients with ST elevation myocardial infarction treated with thrombolysis. RESULTS: After revascularization, global strain and LVEF correlated well with infarct size measured by contrast-enhanced cardiac magnetic resonance. A cutoff value of 15.0% for global strain had a sensitivity of 90% and a specificity of 86% to identify myocardial infarcts larger than 20%. Interobserver variability, expressed by intraclass correlation coefficients, for global strain and LVEF was 0.91 and 0.72, respectively. CONCLUSIONS: LV global strain is a more precise diagnostic predictor of large infarcts compared with LVEF and is more reproducible. Global strain measured after revascularization demonstrates advantages over LVEF in the evaluation of LV injury in patients with ST-elevation myocardial infarction. PMID- 19815384 TI - Aberrant left innominate artery from the left descending aorta in right aortic arch: echocardiographic diagnosis. AB - A right aortic arch with a left descending aorta and an aberrant left innominate artery is a rare but recognized vascular anomaly that can result in compression of the trachea and the esophagus. This vascular anomaly has been diagnosed using cardiac catheterization and angiography. Recently, computed tomography and magnetic resonance imaging have been used for noninvasive diagnosis. The aim of this report is to highlight the possibility of echocardiographic diagnosis. PMID- 19815385 TI - Anomalous origin of the right coronary artery from the left ventricle. AB - A 22-month-old boy was referred to pediatric cardiology for evaluation of a heart murmur. He was asymptomatic except for occasional wheezing with activity. On evaluation, he was found to have both systolic and diastolic murmurs. Electrocardiography demonstrated possible left ventricular hypertrophy, with no evidence of ST-segment abnormalities. Echocardiography and cardiac catheterization showed an anomalous origin of the patient's right coronary artery from his left ventricle, just inferior to his aortic valve annulus. PMID- 19815386 TI - Left ventricle to right ventricle size discrepancy in the fetus: the presence of critical congenital heart disease can be reliably predicted. AB - BACKGROUND: Prenatal ventricular size discrepancy with disproportionately smaller left ventricle than right ventricle (L-R/VD) can be a marker for important left sided structural heart disease in the newborn. METHODS: We reviewed the echocardiograms of all fetuses evaluated at our center with L-R/VD from July 1, 2004 to January 1, 2008. RESULTS: Of the 35 fetuses, 20 (57%) had critical arch obstruction and underwent neonatal intervention (group 1); 15 (43%) did not require newborn intervention (group 2). Ratios comparing left with right heart structures were significantly lower in group 1 fetuses compared with group 2 fetuses. Aortic arch measurement 1.40 or (a history of) intermittent claudication were excluded. Serial troponin-T measurements were performed routinely before and after surgery. The main study endpoint was perioperative myocardial damage, the composite of myocardial ischaemia and infarction. Multivariate regression analyses, adjusted for conventional risk factors, evaluated the relation between asymptomatic low ABI and perioperative myocardial damage. RESULTS: In total, 148 (23%) patients had asymptomatic low ABI (mean 0.73, standard deviation+/-0.13). Perioperative myocardial damage was recorded in 107 (18%) patients. Multivariate regression analyses demonstrated that asymptomatic low ABI was associated with an increased risk of perioperative myocardial damage (odds ratio (OR): 2.4, 95% CI: 1.4-4.2) CONCLUSIONS: This study demonstrated that asymptomatic low ABI has a prognostic value to predict perioperative myocardial damage in vascular surgery patients, incremental to risk factors imbedded in conventional cardiac risk indices. PMID- 19815433 TI - Effect of different penetration enhancers on diclofenac permeation across horse skin. AB - Diclofenac is a hydrophilic non-steroidal anti-inflammatory drug widely used in humans and animals. Previous reports have shown that this compound has low percutaneous absorption in horses. The effect of five penetration enhancers (10% urea, 15% and 20% oleic acid and 5% and 10% d-limonene) on the percutaneous absorption of diclofenac diethylamine through horse skin was evaluated in vitro using Franz-type diffusion cells. All tested penetration enhancers induced a significant increase in diclofenac diethylamine permeation, with limonene showing the highest enhancing effect at the lowest concentration (5%) applied. The presence of the permeation enhancers did not affect lag-time. This is the first in vitro study of the effects of penetration enhancers on transdermal permeation of diclofenac diethylamine across horse skin. The results suggested that urea, limonene and 5% oleic acid were useful for enhancing the transdermal absorption of diclofenac diethylamine and may assist in the development of a transdermal formulation of diclofenac diethylamine for use in horses. PMID- 19815434 TI - The role of N-methyl-D-aspartate receptor subunit NR2B in spinal cord in cancer pain. AB - Cancer pain is one kind of the most common and severe kinds of chronic pain. No breakthrough regarding the mechanisms and therapeutics of cancer pains has yet been achieved. Based on the well established involvement of the NMDA (N-methyl-D aspartate) receptor containing NR2B in inflammatory pain and neuropathic pain and the effective pain relief obtained with ketamine in cancer patients with intractable pain, we supposed that NR2B in the spinal cord was an important factor for cancer pain. In this study, we investigated the possible role of NR2B in the spinal cord using a murine model of bone cancer pain. C3H/HeJ mice were inoculated into the intramedullary space of the right femur with Osteosarcoma NCTC 2472 cells to induce ongoing bone cancer-related pain behaviors. At day 14 after operation, the expression of NR2B mRNA and NR2B protein in the spinal cord were higher in tumor-bearing mice compared to the sham mice. Intrathecal administration of 5 and 10 microg of NR2B subunit-specific NMDA receptor antagonist ifenprodil attenuated cancer-evoked spontaneous pain, thermal hyperalgesia and mechanical allodynia. These results suggest that NR2B in the spinal cord may participate in bone cancer pain in mice, and ifenprodil may be a useful alternative or adjunct therapy for bone cancer pain. The findings may lead to novel strategies for the treatment of bone cancer pain. PMID- 19815435 TI - Pure phase encode magnetic field gradient monitor. AB - Numerous methods have been developed to measure MRI gradient waveforms and k space trajectories. The most promising new strategy appears to be magnetic field monitoring with RF microprobes. Multiple RF microprobes may record the magnetic field evolution associated with a wide variety of imaging pulse sequences. The method involves exciting one or more test samples and measuring the time evolution of magnetization through the FIDs. Two critical problems remain. The gradient waveform duration is limited by the sample T(2)*, while the k-space maxima are limited by gradient dephasing. The method presented is based on pure phase encode FIDs and solves the above two problems in addition to permitting high strength gradient measurement. A small doped water phantom (1-3 mm droplet, T(1), T(2), T(2)* < 100 micros) within a microprobe is excited by a series of closely spaced broadband RF pulses each followed by FID single point acquisition. Two trial gradient waveforms have been chosen to illustrate the technique, neither of which could be measured by the conventional RF microprobe measurement. The first is an extended duration gradient waveform while the other illustrates the new method's ability to measure gradient waveforms with large net area and/or high amplitude. The new method is a point monitor with simple implementation and low cost hardware requirements. PMID- 19815436 TI - Cross-calibration of iDXA and Prodigy on spine and femur scans in Korean adults. AB - In this study, the authors compared bone mineral density (BMD) determined using GE Lunar iDXA and Prodigy and derived cross-calibration equations for the 2 devices in Korean adults. One hundred subjects (66 women and 34 men) participated in this study. Bone mineral density of spine and femur was measured by iDXA and Prodigy dual-energy X-ray absorptiometry (GE Lunar, Madison, WI). Subjects were divided into 3 groups. The first group (30 subjects) was scanned twice using Prodigy for precision testing and then once using iDXA. The second group (30 subjects) was scanned twice using iDXA and then once using Prodigy. Cross calibration equations were derived using these results. The derived equations were tested in the third group (40 subjects). Predicted values from calculations based on Prodigy findings were compared with measured iDXA data. A significant difference was found between the BMD determined using the 2 devices (p < 0.001). However, linear regression analysis showed a high level of agreement between the two (r(2) from 0.984 to 0.994, p < 0.001). Bland-Altman analysis revealed no significant correlations between Prodigy and iDXA. Cross-calibration equations decreased systematic errors between Prodigy and iDXA by 0.4% at the spine, 0.8% at the femoral neck, and 0.1% at the total femur. A high level of agreement was found between Prodigy and iDXA in Korean adults. Cross-calibration equations proved reliable based on comparisons of measured and calculated BMD values. PMID- 19815437 TI - Phalangeal quantitative ultrasound and bone mineral density in evaluating cortical bone loss: a study in postmenopausal women with primary hyperparathyroidism and subclinical iatrogenic hyperthyroidism. AB - Twenty-five postmenopausal women with primary hyperparathyroidism (PHPT) and 30 age-matched women with subclinical hyperthyroidism (sHTH) were studied to assess cortical bone loss. One hundred two healthy women were also recruited. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry at lumbar spine (LS), femoral neck (FN) and femoral total (FT), and at one-third of the radius (R). Amplitude-dependent speed of sound (ADSoS) and Ultrasound Bone Profile Index (UBPI) were also evaluated using phalangeal quantitative ultrasound (QUS). A significant correlation was found between QUS and BMD at LS (ADSoS, p < 0.05) and R (ADSoS and UBPI, p < 0.001) in controls. QUS significantly correlated with BMD at LS, FN (p < 0.01), and FT (p < 0.001) in sHTH. No correlations were found in the PHPT group. Mean T-score values of all parameters were significantly lower in patients compared with controls (p < 0.001); however, they did not differ between PHPT and sHTH patients. T-score of R, ADSoS, and UBPI was reduced compared with other sites (p < 0.001) in both diseases. In postmenopausal women with PHPT and sHTH, bone loss is mainly detectable at cortical level. However, qualitative and/or structural changes of bone could account for the lack of correlations between these 2 techniques at cortical sites. PMID- 19815438 TI - Hereditary 1,25-dihydroxyvitamin D-resistant rickets with alopecia resulting from a novel missense mutation in the DNA-binding domain of the vitamin D receptor. AB - The rare genetic recessive disease, hereditary vitamin D resistant rickets (HVDRR), is caused by mutations in the vitamin D receptor (VDR) that result in resistance to the active hormone 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3) or calcitriol). In this study, we examined the VDR from a young boy with clinical features of HVDRR including severe rickets, hypocalcemia, hypophosphatemia and partial alopecia. The pattern of alopecia was very unusual with areas of total baldness, adjacent to normal hair and regions of scant hair. The child failed to improve on oral calcium and vitamin D therapy but his abnormal chemistries and his bone X-rays normalized with intravenous calcium therapy. We found that the child was homozygous for a unique missense mutation in the VDR gene that converted valine to methionine at amino acid 26 (V26M) in the VDR DNA-binding domain (DBD). The mutant VDR was studied in the patient's cultured skin fibroblasts and found to exhibit normal [(3)H]1,25(OH)(2)D(3) binding and protein expression. However, the fibroblasts were unresponsive to treatment with high concentrations of 1,25(OH)(2)D(3) as demonstrated by their failure to induce CYP24A1 gene expression, a marker of 1,25(OH)(2)D(3) responsiveness. We recreated the V26M mutation in the WT VDR and showed that in transfected COS-7 cells the mutation abolished 1,25(OH)(2)D(3)-mediated transactivation. The mutant VDR exhibited normal ligand-induced binding to RXRalpha and to the coactivator DRIP205. However, the V26M mutation inhibited VDR binding to a consensus vitamin D response element (VDRE). In summary, we have identified a novel V26M mutation in the VDR DBD as the molecular defect in a patient with HVDRR and an unusual pattern of alopecia. PMID- 19815439 TI - Biochemical profiling to predict disease severity in metachromatic leukodystrophy. AB - Metachromatic leukodystrophy is a neurodegenerative disease that is characterized by a deficiency of arylsulfatase A, resulting in the accumulation of sulfatide and other lipids in the lysosomal network of affected cells. Accumulation of sulfatide in the nervous system leads to severe impairment of neurological function with a fatal outcome. Prognosis is often poor unless treatment is carried out before the onset of clinical symptoms. Pre-symptomatic detection of affected individuals may be possible with the introduction of newborn screening programs. The ability to accurately predict clinical phenotype and rate of disease progression in asymptomatic individuals will be essential to assist selection of the most appropriate treatment strategy. Biochemical profiling, incorporating the determination of residual enzyme protein/activity using immune based assays, and metabolite profiling using electrospray ionization-tandem mass spectrometry, was performed on urine and cultured skin fibroblasts from a cohort of patients representing the clinical spectrum of metachromatic leukodystrophy and on unaffected controls. Residual enzyme protein/activity in fibroblasts was able to differentiate unaffected controls, arylsulfatase A pseudo-deficient individuals, pseudo-deficient compound heterozygotes and affected patients. Metachromatic leukodystrophy phenotypes were distinguished by quantification of sulfatide and other secondarily altered lipids in urine and skin fibroblasts; this enabled further differentiation of the late-infantile form of the disorder from the juvenile and adult forms. Prediction of the rate of disease progression for metachromatic leukodystrophy requires a combination of information on genotype, residual arylsulfatase A protein and activity and the measurement of sulfatide and other lipids in urine and cultured skin fibroblasts. PMID- 19815440 TI - Application of oligonucleotide array CGH to the simultaneous detection of a deletion in the nuclear TK2 gene and mtDNA depletion. AB - Thymidine kinase 2 (TK2), encoded by the TK2 gene on chromosome 16q22, is one of the deoxyribonucleoside kinases responsible for the maintenance of mitochondrial deoxyribonucleotide pools. Defects in TK2 mainly cause a myopathic form of the mitochondrial DNA depletion syndrome (MDDS). Currently, only point mutations and small insertions and deletions have been reported in TK2 gene; gross rearrangements of TK2 gene and possible hepatic involvement in patients with TK2 mutations have not been described. We report a non-consanguineous Jordanian family with three deceased siblings due to mtDNA depletion. Sequence analysis of the father detected a heterozygous c.761T>A (p.I254N) mutation in his TK2 gene; however, point mutations in the mother were not detected. Subsequent gene dosage analysis using oligonucleotide array CGH identified an intragenic approximately 5.8-kb deletion encompassing the 5'UTR to intron 2 of her TK2 gene. Sequence analysis confirmed that the deletion spans c.1-495 to c.283-2899 of the TK2 gene (nucleotide 65,136,256-65,142,086 of chromosome 16). Analysis of liver and muscle specimens from one of the deceased infants in this family revealed compound heterozygosity for the paternal point mutation and maternal intragenic deletion. In addition, a significant reduction of the mtDNA content in liver and muscle was detected (10% and 20% of age- and tissue-matched controls, respectively). Prenatal diagnosis was performed in the third pregnancy. The fetus was found to carry both the point mutation and the deletion. This child died 6months after birth due to myopathy. A serum specimen demonstrated elevated liver transaminases in two of the infants from whom results were available. This report expands the mutation spectrum associated with TK2 deficiency. While the myopathic form of MDDS appears to be the main phenotype of TK2 mutations, liver dysfunction may also be a part of the mitochondrial depletion syndrome caused by TK2 gene defects. PMID- 19815441 TI - Axonal neuropathy with unusual clinical course in young Snowshoe cats. AB - Neuropathies in cats are mostly acquired. In comparison to the dog, only very few inherited forms have been described. This case report describes the clinical and diagnostic findings of a suspected inherited feline axonal neuropathy with a very unique clinical course. Two young related male Snowshoe cats were presented with an initially progressive history of recurrent pelvic limb weakness. Electrodiagnostic investigations suggested an axonopathy and muscle-nerve biopsies confirmed axonal degeneration. Over the following 2 years both cats stabilised without any specific treatment, and repeat electrodiagnostic investigations and muscle biopsy in one cat confirmed the tendency for remission. PMID- 19815442 TI - Fetal origins of insulin resistance and the metabolic syndrome: a key role for adipose tissue? AB - For several years now, the epidemiological data have shown an inverse relationship between birth-weight and the development in later life of cardiovascular disease and metabolic disorders. The term "small for gestational age" (SGA) describes a neonate whose birth-weight is two standard deviations (SD) below the reference mean, corrected for gestational age and gender. SGA is associated with increased risks of developing hypertension, insulin resistance and type2 diabetes. However, the association with an atherogenic lipid profile is less clear. Nevertheless, all of the components of the metabolic syndrome are present. Yet, in spite of the large body of data in the literature, the biological mechanisms underlying this association are still unclear. To explain the association, various hypotheses have been proposed, pointing to the role of a detrimental fetal environment or genetic susceptibility, or interaction between the two, and to the particular dynamic changes in adiposity that occur during catch-up growth. However, not only quantitative, but also qualitative, abnormalities of adipose tissue have been observed, suggesting a critical role of this organ in the development of metabolic complications. PMID- 19815443 TI - Differential effect of plasma or erythrocyte AGE-ligands of RAGE on expression of transcripts for receptor isoforms. AB - AIM: Binding of advanced glycation end-products (AGEs) to the receptor for AGEs (RAGE) contributes to diabetic vascular complications. RAGE transcript splicing generates membrane-bound proteins [full-length (FL) and N-truncated (Nt)] and a soluble protein [endogenous secretory (esRAGE)] that may act as a decoy. We tested the effect of AGE-ligands on the regulation of RAGE isoforms and the consequences on red blood cell (RBC) adhesion. METHODS: RAGE isoforms were measured by real-time RT-PCR assay, using a LightCycler System, in human umbilical vein endothelial cells (HUVECs), incubated with either characterized AGEs [Nvarepsilon-(carboxymethyl)lysine human serum albumin (CML-HSA) and methylglyoxal-modified HSA (MG-HSA)] or with RBCs from diabetic patients (DRBCs). Inhibition of RAGE access was achieved by using blocking either anti-RAGE antibodies or recombinant RAGE. Adhesion of DRBCs to endothelium was measured under flow conditions using HUVECs stimulated with MG-HSA or CML-HSA. Antibodies directed to RBC membrane proteins were tested for blocking DRBC adhesion in static conditions. RESULTS: MG-HSA stimulated the expression of membrane-bound RAGE (FL+Nt) and esRAGE transcripts to similar extents, while CML-HSA and DRBC more selectively induced mRNA for FL and Nt-RAGE. Anti-RAGE antibody inhibited the effect of glycated proteins. Stimulation of HUVECs with CML-HSA enhanced DRBC adhesion, while MG-HSA had no effect. CD233 (band 3) was glycated in DRBC membrane, and anti-CD233 antibodies inhibited the adhesion of DRBCs, as did the anti-RAGE and anti-AGE antibodies. CONCLUSIONS: Receptor engagement by distinct AGEs differentially enhances expression of RAGE isoforms and DRBC adhesion. The CML-adduct, by facilitating adhesion, has more deleterious effects than MG derived AGEs. PMID- 19815444 TI - [The cricopharyngeal muscle and the laryngeal nerves: contribution to the functional anatomy of swallowing]. AB - OBJECTIVES: The aim of our study has been to describe the cricopharyngeal muscle and its nerve supply from the recurrent laryngeal nerve and from the superior laryngeal nerve. METHODS: Six normal adult human pharynges (12 sides) obtained from cadavers were studied. Micro-dissections were carried out thanks to a Leica microscope. Measures were obtained with a micro-digital caliber. RESULTS AND CONCLUSION: The morphological feature of the cricopharyngeal muscle (CPM) showed three different muscular bundles: superior oblique muscle fibers are joining the inferior pharyngeal constrictor muscle, horizontal circular muscle fibers are 11 to 12mm high, and 3mm thick, inferior oblique muscle fibers are joining external layer of the superior oesophagus. The recurrent laryngeal nerve through its dorsal branch supplies the posterior part of the muscular bundles of the CPM. The superior laryngeal nerve by its lateral branch supplies the anterior part of the CPM. So this double vagal innervation of the CPM from superior and inferior laryngeal nerves participates in the laryngo-pharyngeal coordination specially for swallowing. PMID- 19815445 TI - [Half a century of human and medical cytogenetics]. AB - In 1956, the number of chromosomes in humans is set at 46; in 1959, the link between a disability (mongolism) and a chromosomal anomaly (the Down syndrome) is established: human and medical cytogenetics were born. Since then, progress has been remarkable: the techniques of chromosomal and molecular cytogenetics can reach a resolution of the size of a single gene with a pangenomic scope. Practical applications are constantly expanded. The clinical impact is significant, from the genetic counselling in constitutional to the targeted therapies. Fifty years later, cytogenetics can be defined as the science which aims to detect chromosomal abnormalities, whether constitutional or acquired, using chromosomal or molecular techniques aiming to study the arrangement of genes in chromosomes, to quantify the number of gene copy and to look for the presence of gene fusion. PMID- 19815447 TI - The effect of levodopa treatment on cerebral hemodynamics in patients with Parkinson's disease: serial transcranial Doppler studies. AB - Levodopa treatment in patients with Parkinson's disease (PD) is known to cause elevation in serum homocysteine levels. We investigated whether this increase in homocysteine level influences cerebral vascular flow velocity and resistance using transcranial Doppler (TCD). This study included 17 patients with de novo PD. Homocysteine levels and TCD parameters at middle cerebral artery were investigated before and after 3 months of levodopa treatment. Correlation analyses were done between changes in homocysteine levels and TCD parameters. After 3 months of levodopa treatment, homocysteine level increased significantly from 13.3mg/dL to 17.0 mg/dL (p < 0.001), but there were no meaningful changes in mean velocity (MV) and pulsatility index (PI). Correlation analysis revealed that the changes in homocysteine level had negative correlation with MV (r = -0.53, p = 0.027) and positive correlation with PI (r = 0.55, p = 0.028). Our study infer that although short-term treatment of levodopa itself does not cause overall alteration of cerebral blood flow velocities and resistances, patients who has greater degree of increased homocysteine level may still be at a risk of developing cerebral vascular stiffness. PMID- 19815448 TI - Neuralgic amyotrophy following botulinum toxin injection. PMID- 19815449 TI - Disparate routes through support: negotiating the sites, stages and support of informal dementia care. AB - Worldwide people with dementia are usually cared for at home by informal carers who may themselves have poor health and/or live in social situations which intensify their needs. The scale of these needs continues to be underappreciated and they are exacerbated by the limited social, cultural and emotional resources that carers can draw upon. This paper looks at the disparities in support, and the complex negotiations made by carers, as they reconcile the everyday realities of informal care in the home. Appreciation of these issues is essential in understanding carers' coping strategies in an ageing population. PMID- 19815450 TI - Building social cognitive models of language change. AB - Studies of language change have begun to contribute to answering several pressing questions in cognitive sciences, including the origins of human language capacity, the social construction of cognition and the mechanisms underlying culture change in general. Here, we describe recent advances within a new emerging framework for the study of language change, one that models such change as an evolutionary process among competing linguistic variants. We argue that a crucial and unifying element of this framework is the use of probabilistic, data driven models both to infer change and to compare competing claims about social and cognitive influences on language change. PMID- 19815446 TI - Parkin and PINK1 parkinsonism may represent nigral mitochondrial cytopathies distinct from Lewy body Parkinson's disease. AB - Recent authors have concluded that Parkinson's disease (PD) is too heterogeneous to still be considered a single discrete disorder. They advise broadening the concept of PD to include genetic parkinsonisms, and discard Lewy pathology as the confirmatory biomarker. However, PD seen in the clinic is more homogeneous than often recognized if viewed from a long-term perspective. With appropriate diagnostic criteria, it is consistently associated with Lewy neuropathology, which should remain the gold standard for PD diagnostic confirmation. PD seen in the clinic has an inexorable course with eventual development of not only levodopa-refractory motor symptoms, but often cognitive dysfunction and prominent dysautonomia. This contrasts with homozygous parkin, PINK1 or DJ1 parkinsonism, characterized by young-onset (usually <40 years), and a comparatively benign course of predominantly levodopa-responsive symptoms without dementia or prominent dysautonomia. Parkin neuropathology is non-Lewy, with neurodegeneration predominantly confined to substantia nigra (and locus ceruleus), consistent with the limited clinical phenotype. Given the restricted and persistently levodopa responsive phenotype, these familial cases might be considered "nigropathies". Based on emerging laboratory evidence linking parkin and PINK1 (and perhaps DJ1) to mitochondrial dysfunction, these nigropathies may represent nigral mitochondrial cytopathies. The dopaminergic substantia nigra is uniquely vulnerable to mitochondrial challenges, which might at least be partially attributable to large energy demands consequent to thin, unmyelinated axons with enormous terminal fields. Although sporadic PD is also associated with mitochondrial dysfunction, Lewy neurodegeneration represents a more pervasive disorder with perhaps a second, or different primary mechanism. PMID- 19815451 TI - Adsorption of difluoromethane on titanium dioxide: investigation of the FTIR spectra and quantum-mechanical studies of the adsorbate-substrate structures. AB - The interaction of difluoromethane (CH(2)F(2)) with the TiO(2) surface (P25 Degussa) at room temperature has been studied by Fourier-transform infrared spectroscopy for the first time. From the comparison between the adsorption characteristics and the gas-phase spectra it can be deduced that the molecule adsorbs through an acid-base interaction between one F atom and the surface Lewis acid site (Ti(4+)) and an H-bond between the CH(2) group and the surface Lewis basic site (OH(-) or O(2-)). In order to obtain more information about the orientation geometry and the variation of the molecular structural parameters, a quantum-mechanical investigation at DFT/B3LYP level has been also performed, considering the anatase (101) surface and focusing on the O(2-) as Lewis basic site. The resulting adsorbate-substrate structures involve the formation of an acid-base interaction between one F atom and the Ti(4+) ion and differ for the number of the involved H-bonds. According to the scaled vibrational frequencies, the simulated adsorption model which better agrees with the experimental data corresponds to that in which the CH(2) group interacts with the surface by only one H-bond. PMID- 19815454 TI - Effect of wrist position on sensorimotor processing in the grip-lift task. PMID- 19815452 TI - Social, organizational, and contextual characteristics of clinical decision support systems for intensive insulin therapy: a literature review and case study. AB - INTRODUCTION: Evaluations of computerized clinical decision support systems (CDSS) typically focus on clinical performance changes and do not include social, organizational, and contextual characteristics explaining use and effectiveness. Studies of CDSS for intensive insulin therapy (IIT) are no exception, and the literature lacks an understanding of effective computer-based IIT implementation and operation. RESULTS: This paper presents (1) a literature review of computer based IIT evaluations through the lens of institutional theory, a discipline from sociology and organization studies, to demonstrate the inconsistent reporting of workflow and care process execution and (2) a single-site case study to illustrate how computer-based IIT requires substantial organizational change and creates additional complexity with unintended consequences including error. DISCUSSION: Computer-based IIT requires organizational commitment and attention to site-specific technology, workflow, and care processes to achieve intensive insulin therapy goals. The complex interaction between clinicians, blood glucose testing devices, and CDSS may contribute to workflow inefficiency and error. Evaluations rarely focus on the perspective of nurses, the primary users of computer-based IIT whose knowledge can potentially lead to process and care improvements. CONCLUSION: This paper addresses a gap in the literature concerning the social, organizational, and contextual characteristics of CDSS in general and for intensive insulin therapy specifically. Additionally, this paper identifies areas for future research to define optimal computer-based IIT process execution: the frequency and effect of manual data entry error of blood glucose values, the frequency and effect of nurse overrides of CDSS insulin dosing recommendations, and comprehensive ethnographic study of CDSS for IIT. PMID- 19815455 TI - A commentary on "Periodic limb movements both in non-REM and in REM sleep: relationships between cerebral and autonomic activities" by Allena et al. Clin Neurophysiol 2009;120:1282-90. PMID- 19815456 TI - Motor evoked potential latency, motor threshold and electric field measurements as indices of transcranial magnetic stimulation depth. PMID- 19815457 TI - Serum collagen-derived peptides are unaffected by physical training in older sedentary subjects. AB - Cardiac fibrosis occurs with normal aging and may have important pathological consequences. Accumulating evidence shows that biochemical assessment of fibrosis using collagen markers such as serum levels of the aminoterminal propeptide of types I and III collagen (PINP, PIIINP) and the carboxyterminal telopeptide of type I collagen (ICTP) represents a practical, validated and non-invasive method to assess myocardial collagen turnover. It is generally accepted that a sedentary lifestyle may be at least partly responsible for the age-related changes in the cardiovascular system. We aimed to investigate whether dynamic aerobic endurance training at lower or higher intensity (33% and 66% of heart rate reserve) may influence markers of collagen synthesis (PINP, PIIINP) and degradation (ICTP) in at least 55-year-old healthy sedentary men and women. A randomised crossover study comprising three 10-week periods was performed. In the first and third period, participants exercised at, respectively, lower or higher intensity in random order, with a sedentary period in between. Training programs were identical except for intensity and were performed three times 50 min/week. 39 (18 men) out of 48 randomised participants completed the study; age averaged 59 years. Serum concentrations of PINP, PIIINP and ICTP were determined at baseline and at the end of each 10-week period. Our results showed that 10 weeks of endurance training at lower or higher intensity did not significantly alter serum markers of collagen synthesis and degradation in healthy older men and women. PMID- 19815458 TI - Using EMGs and kinematics data to study the take-off technique of experts and novices for a pole vaulting short run-up educational exercise. AB - This study attempts to characterise the electromyographic activity and kinematics exhibited during the performance of take-off for a pole vaulting short run-up educational exercise, for different expertise levels. Two groups (experts and novices) participated in this study. Both groups were asked to execute their take off technique for that specific exercise. Among the kinematics variables studied, the knee, hip and ankle angles and the hip and knee angular velocities were significantly different. There were also significant differences in the EMG variables, especially in terms of (i) biceps femoris and gastrocnemius lateralis activity at touchdown and (ii) vastus lateralis and gastrocnemius lateralis activity during take-off. During touchdown, the experts tended to increase the stiffness of the take-off leg to decrease braking. Novices exhibited less stiffness in the take-off leg due to their tendency to maintain a tighter knee angle. Novices also transferred less energy forward during take-off due to lack of contraction in the vastus lateralis, which is known to contribute to forward energy transfers. This study highlights the differences in both groups in terms of muscular and angular control according to the studied variables. Such studies of pole vaulting could be useful to help novices to learn expert's technique. PMID- 19815459 TI - Living with breast cancer, a Lebanese experience. AB - AIM: The purpose of this qualitative phenomenological study is to provide in depth understanding of the experience of Lebanese women living with breast cancer. BACKGROUND: Breast cancer is the most frequently diagnosed cancer in women worldwide. In Lebanon, a country of 4 million people, breast cancer is as well the most widespread type of cancer among Lebanese women. The meaning of cancer diagnosis, the meaning of childbearing and femininity all have cultural bases in Lebanon. The international literature lacks information on how Lebanese women live with breast cancer when compared with women of other cultures. METHOD: The study followed purposeful sampling and saturation principles in which 10 participants with a mean age of 51.3 years were chosen based on their actual knowledge of the phenomenon, and their readiness to share that knowledge. Data were collected between December 2007 and May 2008. All interviews were audio taped and transcribed verbatim. Data were analyzed based on the Utrecht School of Phenomenology. FINDINGS: Four major core themes describing the participants' lived experience emerged from the interviews: Living with losses; living with guilt feeling; living with fears and uncertainty; Living with the need to know and to share that knowledge. CONCLUSION: The experience of Lebanese women with breast cancer revealed distinctive themes not reported by other women from other cultures. The results of this study challenge health care providers and educators to be aware of the difficulties that Lebanese women are facing when they are living with breast cancer. PMID- 19815460 TI - Evaluation of palliative care nursing education seminars. AB - PURPOSE: More than 50 million people die each year around the world. Nurses are crucial in providing care to these individuals and their families as they spend the most time at the bedside with patients and families. Yet many nurses have received little or no education about palliative care. METHODS/SAMPLE: The Open Society Institute (OSI) and the Open Medical Institute (OMI) partnered with End of-Life Nursing Education Consortium (ELNEC) to develop an international nursing palliative care curriculum. This international curriculum was implemented with two training courses held in Salzburg, Austria in October 2006 (n=38) and April 2008 (n=39) representing 22 Eastern European/Central Asian countries. Participants were asked to establish goals in disseminating the palliative care information when they returned to their country. The participants were mentored/followed for a 12-month period to evaluate their palliative care knowledge as well as challenges encountered. KEY RESULTS: The participants provided excellent ratings for the training courses indicating that the courses were stimulating and met their expectations. The 12-month follow-up demonstrated many challenges (i.e., lack of funds, institutional support, fear of death), in advancing palliative care within each participant's setting/country as well as many examples of successful implementation. CONCLUSIONS: There is an urgent need for improved palliative care throughout the world. The ELNEC-International curriculum is designed to address the need for increased palliative care education in nursing. In order to improve the quality of life for those facing life-threatening illnesses around the world, ongoing support is needed for world wide palliative care educational efforts. PMID- 19815461 TI - Transitioning a master's of nursing course from campus to on-line delivery: Lessons learned. AB - Distance education continues to expand in nursing programs to service students in urban as well as rural areas in Canada. This article discusses the author's experience with transitioning a master's course in nursing from a traditional seminar format with 7-8 students to one that included distance students joining the classroom in real-time. The course, 'Nursing in Tertiary Health Care' which covers the fall and winter semesters addresses issues and concepts associated with advanced practice nursing for clients of all ages in tertiary care settings. The associated practicum selected by the student with the assistance of the faculty member provides an opportunity to examine the five domains of the Clinical Nurse Specialist role (consultation, education, research, leadership and practice) as one example of advanced practice nursing. The author has taught this course for 12 years in a traditional seminar format. During 2006-2007, a synchronous on-line version of this course was also offered whereby distance students joined the classroom at the same time as the other students in house at the university. This was done using personal computers from home or office, and a bridging program called 'Bridgit'. Lessons learned in making this transition will be discussed. PMID- 19815462 TI - Evaluation of new vaccines for tuberculosis in the guinea pig model. AB - The guinea pig is a very useful animal model for evaluating new tuberculosis candidate vaccines. In addition to established methods for bacterial load determinations, new technologies are emerging that allow us to specifically evaluate effects of vaccines on the pathology of the disease process and the expression by the host of cell mediated immunity. Limitations to the model include housing and related costs, which often contribute to issue with study design and adequate statistical power, and the use of laboratory strains of Mycobacterium tuberculosis which lack the high virulence and immune evasion properties of newly emerging clinical isolates. PMID- 19815463 TI - Blood transfusion therapy for 41 earthquake casualties. AB - BACKGROUND: We aimed to analyze the efficacy of blood transfusions given to Wen Chuan earthquake casualties. MATERIALS AND METHODS: Forty-one blood transfusions were administered. Laboratory indexes including routine blood tests, electrolyte tests, and blood coagulation functions were recorded. RESULTS: Between 400-6400mL of whole blood and/or erythrocyte suspensions were given to the casualties. The mean volume was 1233mL. The plasma volume infused in 22 cases (1550mL) differed from 200mL to 9600mL. There was no occurrence of transfusion reaction. Erythrocyte and Hct levels increased significantly after transfusion, while electrolyte levels remained the same. CONCLUSION: Blood transfusion was an effective therapy for the earthquake casualties. PMID- 19815464 TI - On "A comment on 'Prediction of protein structural classes by a new measure of information discrepancy"'. AB - This note points out that the comment article by Kedarisetti et al. (2006a, refer to 'K-comment' hereafter) was based on the code analysis with methodological and mathematical errors. The incorrect modification of the original code and FDOD method has resulted in the wrong conclusions in K-comment. PMID- 19815465 TI - Binasal visual field defects are not specific to vigabatrin. AB - This study investigated the visual defects associated with the antiepileptic drug vigabatrin (VGB). Two hundred four people with epilepsy were grouped on the basis of antiepileptic drug therapy (current, previous, or no exposure to VGB). Groups were matched with respect to age, gender, and seizure frequency. All patients underwent objective assessment of electrophysiological function (wide-field multifocal electroretinography) and conventional visual field testing (static perimetry). Bilateral visual field constriction was observed in 59% of patients currently taking VGB, 43% of patients who previously took VGB, and 24% of patients with no exposure to VGB. Assessment of retinal function revealed abnormal responses in 48% of current VGB users and 22% of prior VGB users, but in none of the patients without previous exposure to VGB. Bilateral visual field abnormalities are common in the treated epilepsy population, irrespective of drug history. Assessment by conventional static perimetry may neither be sufficiently sensitive nor specific to reliably identify retinal toxicity associated with VGB. PMID- 19815466 TI - Efficacy and safety of Creon 24,000 in subjects with exocrine pancreatic insufficiency due to cystic fibrosis. AB - BACKGROUND: Pancreatic enzyme replacement therapy is critical for adequate nutrition in cystic fibrosis (CF) patients with exocrine pancreatic insufficiency (EPI). METHODS: This was a double-blind, randomised, placebo-controlled, two period crossover study assessing efficacy and safety of Creon 24,000-unit capsules in CF subjects > or =12 years with EPI. Patients were randomised to one of two 5-day sequences, Creon/placebo or placebo/Creon (target dose, 4000 lipase units/g fat). Primary outcome was the coefficient of fat absorption (CFA); secondary outcomes were coefficient of nitrogen absorption (CNA), symptoms, and safety. RESULTS: Thirty-two subjects were randomised. Mean CFA and CNA were significantly greater with Creon than placebo (CFA, 88.6% vs. 49.6%; CNA, 85.1% vs. 49.9%; p<0.001 for both). Symptoms were improved and fewer treatment-emergent adverse events were reported with Creon than placebo. One patient discontinued for weight loss unrelated to study drug. CONCLUSIONS: This study demonstrated Creon was effective in treating EPI due to CF and was safe and well tolerated. PMID- 19815467 TI - A novel fractionation method prior to MS-based proteomics analysis using cascade biomimetic affinity chromatography. AB - This is the first report that combines cascade biomimetic affinity fractionation with MS-based proteomics analysis. Our lab has constructed an affinity ligand library composed of thousands of ligands with different protein-binding properties. Structural differences between these ligands result in different non bonded protein-ligand interactions, thus each ligand exhibits a specific affinity to some protein groups. In this work, we first screened out three affinity ligands with large difference in protein-binding properties. Next, cascade combination of these ligands was applied to fractionate tissue sample into simple subgroups prior to trypsin digestion and LC-MS/MS analysis. In this study, 391 non-redundant protein groups were identified in unfractionated rat liver cytosol, 499 protein groups were identified in 2 fractions of the first affinity fractionation, 616 in 4 fractions of the second fractionation, and 738 in 8 fractions of the third fractionation (an 88.74% increase). Ultimately, a total of 859 unique protein groups were identified in all cascade fractions (a 119.6% increase compared with unfractionated sample). The proteins detected in each fraction were bioinformatically categorized according to their physicochemical characteristics (relative molecular mass, pI, GRAVY value and TM helices). This approach highlighted the sensitivity of this method to a wide variety of protein classes. PMID- 19815468 TI - A metabonomic approach identifies human urinary phenylacetylglutamine as a novel marker of interstitial cystitis. AB - An ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) based metabonomic approach was applied to identify a candidate metabolite with not known to be associated with interstitial cystitis (IC). IC is a chronic clinical syndrome associated with urinary frequency and urgency and/or pelvic pain. The ability to non-invasively diagnose the early stage of IC would be important for improving the patient's quality of life. The current standard IC diagnosis is cystoscopy, which is invasive and painful. Urine samples from the following were taken and analyzed: 10 IC patients, 10 bacterial cystitis (BC) patients, and 10 healthy volunteers (HVs) to identify an IC marker; and subsequently analyzed 5 IC patients and 5 HVs for marker validation. The urinary marker of IC was identified as phenylacetylglutamine (PAGN) using NMR and MS/MS analysis. In addition, quantitative methods were developed to determining the urinary PAGN levels using UPLC-UV. The urinary level of PAGN measured relative to creatinine (Cr) was significantly elevated in IC patients (mean 0.47mg/mg Cr) compared with BC patients (mean 0.25mg/mg Cr) and HVs (mean 0.11mg/mg Cr). Interestingly, urinary PAGN/Cr ratios in patients with mild IC (grade I) and moderate IC (grade II) were higher than for patients with severe IC (grade III). Moreover, urinary PAGN/Cr ratios with mild and moderate IC patients (mean 0.30mg/mg Cr) were higher than HVs (mean 0.059mg/mg Cr), in the validation set. These findings establish urinary PAGN/Cr ratios as a novel urinary marker of IC, and may contribute to early diagnosis of IC patients. PMID- 19815469 TI - [Consensus document from the Spanish Society of Paediatric Infectious Diseases (SEIP) on the diagnosis and treatment of congenital cytomegalovirus infection]. AB - Cytomegalovirus (CMV) is the leading cause of congenital infection in developed countries, affecting 0.3 to 0.6% of all live births in Europe. Primary CMV infection occurs in 1 to 4% of seronegative women during pregnancy and may be transmitted to the fetus in 40% of cases. Up to 10% of intrauterine CMV infections result in symptomatic congenital disease at birth. Half of these children and 13% of those born with asymptomatic infection will develop long-term sequelae, especially neurosensory hearing loss and mental retardation. Accurate diagnosis of primary maternal and fetal infection is now possible using the avidity index of anti-CMV IgG and virological testing to detect the virus in amniotic fluid. Symptomatic congenital infection may be preventable using CMV hyperimmune globulin during pregnancy. The gold standard for diagnosis of congenital CMV infection is the detection of the virus in urine within the first 2 weeks of life by rapid cell culture techniques (shell vial) or nucleic acid amplification of viral DNA (PCR). Retrospective diagnosis can be achieved by detection of viral DNA by PCR in dried blood spots (Guthrie card) collected on filter paper in the first days of life. Currently available drugs for the treatment of congenital CMV include ganciclovir and its oral prodrug valganciclovir. Treatment with intravenous ganciclovir for six weeks may prevent hearing deterioration in children with symptomatic congenital CMV infection and central nervous system involvement. Valganciclovir may be an excellent alternative because of its good bio-availability, providing plasma concentrations similar to those achieved with intravenous ganciclovir. PMID- 19815470 TI - [Intrasellar enchondroma in female child with Ollier disease]. PMID- 19815471 TI - [Foreign body management. Combined bronchoscopy in a Paediatric Pulmonology Unit]. AB - The foreign body (FB) aspiration is a significant of morbidity and mortality in childhood. The severity of the acute obstruction of the upper airway and the complications that the FB creates in the lower airway requires early diagnosis and treatment. Classically, the FB is removed with a rigid bronchoscope, with general anaesthesia, by surgeons. Nowadays, there are publications on removing FB with flexible bronchoscopy. There is now discussion on which is the best instrument, rigid or flexible bronchoscope. Flexible bronchoscopy is a minimal invasive procedure that allows the diagnosis to be made and locate the FB, which can only be removed in selected cases. Therefore, authors believe that FB removal is more effective with rigid bronchoscopy. It guarantees patient safety and the success of the procedure. However, both, flexible and rigid bronchoscopes are complementary, so their combined use is the most appropriate choice. We report the management of five cases of FB in paediatric patients with a combined procedure using flexible and rigid bronchoscopy. PMID- 19815472 TI - [Talking about death to children with cancer]. AB - INTRODUCTION: Paediatric cancer treatment includes a multidisciplinary intervention in all treatment phases, and particularly in the palliative phase. One of the main skills is information. This study tries to explore the level of information that children on palliative care have about their own death. SAMPLE AND METHODS: We retrospectively collected the psychosocial variables of 45 oncology patients who died as inpatients in the Hospital Nino Jesus Hospital (HNJS) between 2006 and 2007. The concept of death is analysed according to each child development stage. RESULTS: We found a relationship between the age of the children and the information they have about their own death, as well as a statistical significance between the information that the child has and the information received from their parents. Children between 3 to 6 years old have more information about their own death than children between 7 to 11 years old. CONCLUSIONS: Our results confirmed that the older age group had more information on their imminent death. However children between 3 to 6 years old have more information than children between 7 to 11 years old. Probably as children between 3 to 6 years old have a magical concept of death, it makes it easier to talk about their terminal phase. Over-protection and the difficulty to talk about death shows differences between what children know and what parents tell them about their palliative phase. PMID- 19815473 TI - [Adolescent obesity: anxiety, cognitive and behavioural symptoms characteristic of eating disorders]. AB - OBJECTIVE: To assess the presence of symptoms of anxiety, cognitive and behavioural symptoms characteristic of eating disorders in overweight adolescents. MATERIAL AND METHODS: The sample consisted of 297 adolescents (55.2% female and 44.8% male) aged from 11 to 17 years, divided into four groups according to their BMI percentile: normal weight, overweight, obesity and severe obesity. The questionnaires used were the Eating Disorders Inventory-2 (EDI-2) and the state-trait anxiety questionnaire (STAI). RESULTS: The results showed that the adolescents with severe obesity were more concerned about their physical appearance (body dissatisfaction and obsession with being thin), by their social environment (interpersonal distrust), and showed more anxiety state and anxiety trait than adolescent overweight or obese. In the clinical sample, the adolescents with normal weight showed specific factors for developing eating disorders in the future. CONCLUSIONS: As the BMI increased, symptoms of anxiety and symptoms associated with eating disorders were also increased. The findings of this study are discussed in order to improve therapeutic interventions. PMID- 19815474 TI - [Severe bronchiolitis and anisocoria in a 3-month old infant]. PMID- 19815475 TI - Disordered eating behaviors and body image in a longitudinal pilot study of adolescent girls: what happens 2 years later? AB - We assessed the prospective association of risk factors for eating and body image disturbances after a 2-year follow-up in a community sample of Spanish adolescent girls. The participants included 128 Spanish girls aged 12-14, who took part in a 28-month prospective study. Aspects assessed were eating attitudes (Eating Attitudes Test), influence of the body shape model (questionnaire on influences of the aesthetic body shape model), extreme weight-control behaviors (Eating Disorder Examination-Questionnaire), body image (Body Image Questionnaire) and Body Mass Index (BMI). BMI, extreme weight-control behaviors and body image problems emerged as potential predictors of an increase in eating disturbances. An increased influence of the thinness model was significantly associated with reduced body satisfaction and body image problems. Preventive programs are needed to contribute reducing the impact of sociocultural influences with regard to thinness, the use of extreme weight-control behaviors and overweight in adolescents. PMID- 19815476 TI - The drive for muscularity in men: media influences and objectification theory. AB - Presently, objectification theory has yielded mixed results when utilized to explain body image concerns in men. An online survey assessing internalization of media ideals, self-objectification, body surveillance, body shame, the drive for muscularity, and body mass index (BMI) was completed by 244 predominantly college aged males. Path analyses were used to investigate relationships among these variables where it was hypothesized that objectification variables would mediate the relationship between internalization of media ideals and the drive for muscularity. Internalization of media ideals was the strongest predictor of the drive for muscularity, followed by BMI, though variables of objectification theory had no impact on the drive for muscularity contrary to hypotheses. The results suggest that objectification theory may not be applicable to men as it is currently measured. PMID- 19815477 TI - The influence of the hijab (Islamic head-cover) on perceptions of women's attractiveness and intelligence. AB - This study examined the effects of wearing the hijab, or Islamic headwear, on men's perceptions of women's attractiveness and intelligence. A total of 57 non Muslim men and 41 Muslim men rated a series of images of women, half of whom were unveiled and half of whom wore the hijab. For attractiveness and intelligence ratings, a mixed analysis of variance showed a significant effect of hijab status, with women wearing the hijab being rated more negatively than unveiled women. For attractiveness ratings, there was no significant effect of participant religion, although non-Muslim men rated unveiled women significantly higher than veiled women. For intelligence ratings, non-Muslim men provided significantly higher ratings than Muslim men for both conditions. In addition, Muslim men's ratings of the attractiveness and intelligence of women wearing the hijab was positively correlated with self-reported religiosity. These results are discussed in relation to religious stereotyping within increasingly multi-cultural societies. PMID- 19815478 TI - Necrotising fasciitis of the extremity. AB - Necrotising fasciitis is a rare, life-threatening infection where there is necrosis of fascia, subcutaneous tissues and skin. Three adult patients presented with this condition affecting the extremities and all had predisposing medical factors. Each one complained of severe pain that seemed disproportionate to the skin affected. They underwent prompt surgical debridement and received antibiotic treatment and supportive therapy. These cases highlight the increasing incidence and differences in presentation of necrotising fasciitis, as well as the important symptom of pain. Patients can become moribund and require intensive care support. High mortality rates are well recognised. PMID- 19815479 TI - Surgery plus targeted therapy for renal cell carcinoma with isolated spermatic cord metastasis. AB - We present a case of renal cell carcinoma with metastasis to the ipsilateral spermatic cord. We report the clinical presentation and management of this case and review the scientific literature of this unusual case presentation. PMID- 19815480 TI - Tolerance of sunitinib in dialyzed patients with metastatic renal cell carcinoma. AB - The use of sunitinib in dialysis patients is poorly described but is of clinical importance. We report 2 cases of patients receiving sunitinib for metastatic renal cell carcinoma while undergoing dialysis. The first patient is undergoing hemodialysis and, though responding to sunitinib, is having significant fatigue and hypertension. The second patient underwent peritoneal dialysis and also had significant problems with hypertension. The tolerance of sunitinib in the setting of dialysis can be challenging as these interventions can have synergistic side effects. Close monitoring for toxicity and dosage manipulations might be required if such therapy is attempted. PMID- 19815482 TI - Dose escalation using a hypofractionated, intensity-modulated radiation therapy boost for localized prostate cancer: preliminary results addressing concerns of high or low alpha/beta ratio. AB - PURPOSE: The possibility that prostate cancers have a low alpha/beta ratio led to a schedule including a hypofractionated boost. The purpose of this study was to analyze the outcomes of this regimen. PATIENTS AND METHODS: Between 2002 and 2007, 125 patients with localized prostate cancer were treated. Median follow-up was 33 months. Radiation therapy was delivered to a planning target volume including the prostate and seminal vesicles with a 1-1.5 cm margin to block edge using a 6-field technique to 45 Gy in 25 fractions. This was followed by a 2.5 Gy/fraction intensity-modulated radiation therapy boost to the prostate alone to a total dose of 75 Gy in 61 low-risk patients and 77.5 Gy to the prostate and seminal vesicles in 64 high- and intermediate-risk patients. RESULTS: There have been 2 (1.6%) biochemical failures, 1 death from prostate cancer, and 1 death in a patient with no evidence of disease. Rates of acute genitourinary and gastrointestinal toxicity (grade 1 and 2) for the whole group were 31.2% and 16%, respectively. Rates of chronic genitourinary and gastrointestinal toxicity (grade 1 and 2) for the whole group were 30.4% and 27.2%, respectively. There were 2 patients (1.6%) with grade 3 gastrointestinal toxicity at 12 and 18 months' follow-up. They had radiation proctitis requiring laser cauterization. CONCLUSION: The preliminary results of this novel schedule were excellent. Given that the alpha/beta ratio is still in question, this technique addresses concerns regarding low and high ratios. This technique is a suitable alternative method of dose escalation in the treatment of localized prostate cancer. PMID- 19815483 TI - Safety and efficacy of PF-3512676 for the treatment of stage IV renal cell carcinoma: an open-label, multicenter phase I/II study. AB - PURPOSE: Single-agent PF-3512676 (agatolimod), a Toll-like receptor 9 agonist, was examined in an open-label, single-arm, multicenter phase I/II study to determine its maximum tolerated dose (MTD), safety profile, antitumor activity, pharmacokinetics, and immunologic effects in patients with advanced metastatic renal cell carcinoma (RCC). PATIENTS AND METHODS: PF-3512676 was administered subcutaneously weekly for up to 24 weeks to 39 adults with stage IV RCC. Patients were excluded if they had received previous therapy other than surgery. Phase I dose escalation began at 0.08 mg/kg, with phase II expansion to 20 patients to estimate objective response rates at 0.16 mg/kg. Doses were subsequently escalated to 0.81 mg/kg according to the phase I design. RESULTS: An MTD was not reached. One patient who received 0.54 mg/kg had dose-limiting toxicities (grade 3 nonhematologic adverse events [AEs], including anorexia). The most commonly reported AEs were flu-like symptoms and local injection-site reactions of mild-to moderate severity. The most commonly reported serious AE was grade 3 fatigue in 4 patients (10%). Grade 4 AEs included anemia, exacerbated dyspnea, and polyarthralgia in 1 patient each. Two patients (5%), 1 each in the 0.16-mg/kg and 0.54-mg/kg cohorts, achieved a partial response. Both responses were durable (35 and 40 months). CONCLUSION: This was the first study to examine PF-3512676 safety and antitumor activity in patients with advanced RCC. Single-agent treatment was tolerable. At the doses tested, PF-3512676 had modest antitumor activity. Additional studies in combination with other agents or at higher monotherapy doses might be warranted. PMID- 19815484 TI - Arsenic trioxide in recurrent urothelial cancer: a cancer and leukemia group B phase II trial (CALGB 99903). AB - BACKGROUND: Arsenic trioxide is highly active in patients with acute promyelocytic leukemia. There are also preclinical data to suggest that this drug might be active in nonhematopoietic malignancies, and transitional cell carcinoma cell lines are particularly sensitive to this agent. PATIENTS AND METHODS: Twelve evaluable patients with metastatic urothelial cancer were treated with arsenic trioxide in a phase II trial conducted by the Cancer and Leukemia Group B. Eligible patients were required to have measurable urothelial cancer and a maximum of 1 previous chemotherapy regimen. Arsenic trioxide was given at a dose of 0.3 mg/kg daily for 5 days every 28 days. RESULTS: No major responses were observed; 4 patients achieved stable disease. The median survival was 6.5 months (95% CI, 3.9-13.4 months). The most commonly observed toxicities included fatigue and malaise, anemia, nausea, emesis, and constipation. CONCLUSION: Arsenic trioxide at this dose and schedule does not have significant activity in previously treated urothelial cancer and has substantial toxicity in this patient population. PMID- 19815485 TI - The mechanism of action of estrogen in castration-resistant prostate cancer: clues from hormone levels. AB - BACKGROUND: Estrogen therapy plays a role in the management of castration resistant prostate cancer (CRPC), although the mechanism of action is not fully known. This current analysis reports the relationship of change in adrenal androgen levels and prostate-specific antigen (PSA) response in patients with CRPC treated with estrogen therapy. PATIENTS AND METHODS: Hormone levels were measured for patients with CRPC treated in a multicenter phase II trial of 2 estrogen-containing compounds, the herbal supplement PC-SPES and diethylstilbestrol (DES), with known efficacy in CRPC. Patients with castrate levels of testosterone were randomized to initially receive either PC-SPES 960 mg t.i.d. or DES 3 mg/day. Levels of testosterone, dihydrotestosterone (DHT), estradiol, estrone, sex hormone-binding globulin (SHBG), dehydroepiandrosterone (DHEA), DHEA-sulfate (DHEA-S), and androstenedione were obtained at baseline and at 12-week intervals until disease progression. Hormone levels were obtained for 38 patients, 20 treated with PC-SPES and 18 treated with DES. RESULTS: Significant declines between baseline and 12 weeks of treatment were observed in levels of serum testosterone (P < .001), estrone (P = .02), and DHEA (P < .001). The percent changes at 12 weeks in these hormone levels were inversely proportional to baseline values as measured by Spearman's rank correlation (testosterone: -0.41, P = .01; estrone: -0.64, P = .0001; DHEA: -0.39, P = .02). Levels of SHBG increased in almost all of the patients (97%), with a median percent increase of > 5-fold (P < .0001). Of the 38 evaluable patients, 15 (39% [95% CI, 24%-57%]) experienced a > 50% decline in PSA level. There was no significant difference between treatment groups or between responders and nonresponders in baseline distributions for any of the hormones. At follow-up, 73% of the responders had a decline in the level of DHEA-S compared with 41% of the nonresponders, resulting in a difference in the distribution of the percent change between the subsets (Mann-Whitney test: P = .03). Conversely, 64% of the responders compared with 30% of the nonresponders experienced an increase in DHT, with differing distributions of percent change (P = .02). CONCLUSION: Androgens decline in response to estrogen therapy. A decline in DHEA-S and a rise in DHT are both associated with a decline in PSA while patients receive estrogen therapy. PMID- 19815486 TI - Anti-epidermal growth factor receptor monoclonal antibody cetuximab plus Doxorubicin in the treatment of metastatic castration-resistant prostate cancer. AB - PURPOSE: An open-label, dose-escalating phase Ib/IIa trial was performed to establish a safety profile of ascending doses of cetuximab (IMC C225) in combination with doxorubicin administered weekly for 6 treatments in patients with metastatic castration-resistant prostate cancer. The secondary endpoint was to assess the efficacy of cetuximab in combination with doxorubicin as well as to determine the optimal biologic dose and the maximum tolerated dose. PATIENTS AND METHODS: Patients in 8 groups received escalating doses of cetuximab 20-300 mg/m(2) plus doxorubicin 15 or 20 mg/m(2) given intravenously weekly for 6 consecutive weeks, followed by a 1-week observation period. A treatment response was defined as a > 50% decline in prostate-specific antigen (PSA) or regression of radiographically measurable disease. RESULTS: Of the 36 treated patients, 25% had grade 2 neutropenia, 39% had leukopenia, and 44% had stomatitis at doxorubicin 20 mg/m(2). Erythematous skin exanthema was seen in 38% of the patients. There was no significant regression of bone or soft tissue disease, but stable disease was observed in 20 (65%) of the 31 patients with bone disease and 14 (61%) of the 23 patients with lymph node disease. Declines in PSA were modest in the 36 patients, with 1 (2.7%) with an 80% decline from baseline, 2 (5.6%) with > 50% to < 80% declines, and 14 (39%) with progression. Median survival was approximately 18 months. CONCLUSION: In a heavily pretreated population of men with metastatic castration-resistant prostate cancer, this study of cetuximab/doxorubicin was associated with minimal PSA declines posttherapy, though median survival was longer compared to historical control groups. Further studies with cetuximab combined with more contemporary chemotherapy for castration-resistant prostate cancer might be warranted. PMID- 19815487 TI - Drug management of prostate cancer: prevalence and consequences of renal insufficiency. AB - BACKGROUND: The Renal Insufficiency and Anticancer Medications (IRMA) study reported a renal insufficiency (RI) prevalence of 50%-60% in a population of almost 5000 patients with solid tumors, 80% of whom were being treated with anticancer drugs that either necessitated dosage adjustment or were potentially nephrotoxic drugs. A national multicenter study from 15 cancer centers in France analyzed IRMA data on patients with prostate cancer. PATIENTS AND METHODS: Data on patients with prostate cancer from the IRMA study were analyzed. Renal function was calculated using Cockcroft-Gault and abbreviated Modification of Diet in Renal Disease (aMDRD) formulas to estimate the prevalence of RI. Anticancer drugs' potential renal toxicity and need for dosage adjustment were detailed. RESULTS: Of the 222 IRMA patients with prostate cancer, 14.9% had a serum creatinine (SCr) level of > 110 micromol/L. When using Cockcroft-Gault and aMDRD formulas, 62.6% and 55.9%, respectively, of the patients had RI. Of the 228 anticancer drug prescriptions, 82.9% required dose adjustments for RI or were drugs with no available data on their administration in patients with RI. Of the patients treated, 86.9% received >or= 1 such drug, but only 29.1% received nephrotoxic drugs. CONCLUSION: The prevalence of RI in patients with prostate cancer was very high in spite of a normal SCr level in most cases. Some anticancer drugs, particularly some bisphosphonates and platinum salts, might be nephrotoxic and/or need dosage adjustment. However, other important drugs in prostate cancer, such as docetaxel, neither require dose reduction nor present with potential nephrotoxicity. Both issues were significantly more important in the patients with bone metastases compared with those with nonmetastatic disease. PMID- 19815488 TI - Activity of dutasteride plus ketoconazole in castration-refractory prostate cancer after progression on ketoconazole alone. AB - BACKGROUND: Ketoconazole is a commonly used secondary hormonal therapy in castration-refractory prostate cancer (CRPC), but disease progression inevitably occurs. Both prostatic and metastatic lesions in patients with CRPC overexpress 5 alpha reductase (SRDA5) type I. We hypothesized that SRDA5 inhibition in combination with ketoconazole would mitigate progression after treatment with ketoconazole alone. PATIENTS AND METHODS: A total of 10 patients with CRPC with progression after ketoconazole treatment were treated with a combination of ketoconazole plus dutasteride 0.5 mg/day, a dual SRDA5 inhibitor. RESULTS: After dutasteride addition, 8 (80%) of the 10 patients had varying degrees of prostate specific antigen (PSA) decline relative to baseline. Median progression-free survival after dutasteride addition was 4.9 months (range, 2.7+ to 9.8 months); no patient had a >OR= 50% PSA decline. CONCLUSION: We conclude that dutasteride added to ketoconazole at the time progression might prolong time to PSA progression in patients with CRPC. PMID- 19815489 TI - A case of adult metastatic Xp11 translocation renal cell carcinoma treated successfully with sunitinib. AB - Renal cell carcinomas (RCC) associated with Xp11.2 translocations (transcription factor E3 gene fusions) are rare tumors predominantly reported in children. Although only a small number of adult Xp11.2 translocation RCC cases have been described, this is likely due to the possibility of this tumor being unrecognized because of its histologic similarity to either clear cell or papillary subtypes based on hematoxylin and eosin staining. Herein, we describe the first successful case of systemic treatment of metastatic Xp11 RCC with sunitinib, a currently accepted standard of care in metastatic RCC, and attempt to elucidate potential mechanisms behind sunitinib activity in this histologic subtype. PMID- 19815490 TI - Primary extraosseous ewing sarcoma of the kidney with level III inferior vena cava thrombus. AB - Ewing sarcoma typically presents as a skeletal-based tumor, with rare instances of peripheral primitive neuroectodermal tumor (PNET) arising in the soft tissues. Few examples of organ-based PNETs have been previously described in the literature. These tumors are exceedingly rare as a primary renal neoplasm. To our knowledge, this is the first report documenting a primary extraskeletal Ewing sarcoma of the kidney with thrombus extending into the intrahepatic inferior vena cava thrombus (level III). PMID- 19815491 TI - A renal mass in a patient with melanoma. AB - A 61-year-old man presented to the emergency room with significant weight loss. Laboratory analysis revealed elevations in blood urea nitrogen, creatinine, and white blood cell count. Computed tomography imaging showed a large, infiltrative mass in the right renal vein, with metastasis to the brain. Biopsy of soft tissue mass and kidney revealed positive staining for malignant melanoma. Malignant melanoma to the kidney is extremely rare, and imaging modalities alone cannot differentiate neoplasms in the kidney. It is therefore necessary to use specific immunocytochemical staining along with imaging modalities to make a specific diagnosis when the primary origin of the tumor is unknown. PMID- 19815492 TI - Sociology: a lost connection in social psychology. AB - For the first half of the 20th century, sociology was one of the closest allies of social psychology. Over the past four decades, however, the connection with sociology has weakened, whereas new connections with neighboring disciplines (e.g., biology, economics, political science) have formed. Along the way, the sociological perspective has been largely lost in mainstream social psychology in the United States. Most social psychologists today are not concerned with collective phenomena and do not investigate social structural factors (e.g., residential mobility, socioeconomic status, dominant religion, political systems). Even when the social structural factors are included in the analysis, psychologists typically treat them as individual difference variables. Sociologist C. Wright Mills famously promoted sociological imagination, or the ability to see distal yet important social forces operating in a larger societal context. By comparing sociological perspectives to psychological perspectives, this article highlights the insights that the sociological perspective and sociological imagination can bring to social psychology. PMID- 19815494 TI - Effectiveness of adalimumab in treating patients with active psoriatic arthritis and predictors of good clinical responses for arthritis, skin and nail lesions. AB - OBJECTIVES: To evaluate the effectiveness of adalimumab in patients with psoriatic arthritis (PsA) and identify predictors of good clinical response for joint and skin lesions. METHODS: Patients received adalimumab 40 mg every other week in addition to standard therapy in this prospective, 12-week, open-label, uncontrolled study. Four definitions of good clinical response were used: > or =50% improvement in American College of Rheumatology response criteria (ACR50), good response according to European League Against Rheumatism (EULAR) guidelines, a > or =3-grade improvement in Physician Global Assessment of psoriasis (PGA) and a > or =50% improvement in the Nail Psoriasis Severity Index (NAPSI). Response predictors were determined by logistic regression with backward elimination (selection level was 5%). RESULTS: Of 442 patients, 94% completed 12 weeks of treatment. At week 12, 74%, 51% and 32% of the patients had achieved ACR20, 50 and 70, respectively; 87% and 61% experienced moderate and good responses according to EULAR criteria, respectively. The percentage of patients with PGA results of "clear/almost clear" increased from 34% (baseline) to 68%. The mean NAPSI score was reduced by 44%. No new safety signals were detected. A lower Health Assessment Questionnaire Disability Index (HAQ-DI) score, greater pain assessment, male sex and absence of systemic glucocorticoid therapy were strongly associated with achievement of ACR50 and good response according to EULAR criteria. In addition, greater C-reactive protein concentration and polyarthritis predicted ACR50, and non-involvement of large joints predicted a good response according to EULAR criteria. CONCLUSIONS: Adalimumab was effective in patients with PsA. Lower impairment of physical function, greater pain, male sex and no systemic treatment with glucocorticoids were factors that increased the chance of achieving a good clinical response. PMID- 19815495 TI - Novel expression signatures identified by transcriptional analysis of separated leucocyte subsets in systemic lupus erythematosus and vasculitis. AB - OBJECTIVE: To optimise a strategy for identifying gene expression signatures differentiating systemic lupus erythematosus (SLE) and antineutrophil cytoplasmic antibody-associated vasculitis that provide insight into disease pathogenesis and identify biomarkers. METHODS: 44 vasculitis patients, 13 SLE patients and 25 age and sex-matched controls were enrolled. CD4 and CD8 T cells, B cells, monocytes and neutrophils were isolated from each patient and, together with unseparated peripheral blood mononuclear cells (PBMC), were hybridised to spotted oligonucleotide microarrays. RESULTS: Using expression data obtained from purified cells a substantial number of differentially expressed genes were identified that were not detectable in the analysis of PBMC. Analysis of purified T cells identified a SLE-associated, CD4 T-cell signature consistent with type 1 interferon signalling driving the generation and survival of tissue homing T cells and thereby contributing to disease pathogenesis. Moreover, hierarchical clustering using expression data from purified monocytes provided significantly improved discrimination between the patient groups than that obtained using PBMC data, presumably because the differentially expressed genes reflect genuine differences in processes underlying disease pathogenesis. CONCLUSION: Analysis of leucocyte subsets enabled the identification of gene signatures of both pathogenic relevance and with better disease discrimination than those identified in PBMC. This approach thus provides substantial advantages in the search for diagnostic and prognostic biomarkers in autoimmune disease. PMID- 19815496 TI - Transgenic songbirds offer an opportunity to develop a genetic model for vocal learning. AB - Zebra finches are widely used for studying the basic biology of vocal learning. The inability to introduce genetic modifications in these animals has substantially limited studies on the molecular biology of this behavior, however. We used an HIV-based lentivirus to produce germline transgenic zebra finches. The lentivirus encoded the GFP regulated by the human ubiquitin-C promoter [Lois C, Hong EJ, Pease S, Brown EJ, Baltimore D (2002) Science 295:868-872], which is active in a wide variety of cells. The virus was injected into the very early embryo (blastodisc stage) to target the primordial germline cells that later give rise to sperm and eggs. A total of 265 fertile eggs were injected with virus, and 35 hatched (13%); 23 of these potential founders (F0) were bred, and three (13%) produced germline transgenic hatchlings that expressed the GFP protein (F1). Two of these three founders (F0) have produced transgenic young at a rate of 12% and the third at a rate of 6%. Furthermore, two of the F1 generation transgenics have since reproduced, one having five offspring (all GFP positive) and the other four offsping (one GFP positive). PMID- 19815497 TI - High-throughput in vivo screening of targeted molecular imaging agents. AB - The rapid development and translation of targeted molecular imaging agents from bench to bedside is currently a slow process, with a clear bottleneck between the discovery of new compounds and the development of an appropriate molecular imaging agent. The ability to identify promising new molecular imaging agents, as well as failures, much earlier in the development process using high-throughput screening techniques could save significant time and money. This work combines the advantages of combinatorial chemistry, site-specific solid-phase radiolabeling, and in vivo imaging for the rapid screening of molecular imaging agents. A one-bead-one-compound library was prepared and evaluated in vitro, leading to the identification of 42 promising lead peptides. Over 11 consecutive days, these peptides, along with a control peptide, were successfully radiolabeled with 4-[(18)F]fluorobenzoic acid and evaluated in vivo using microPET. Four peptides were radiolabeled per day, followed by simultaneous injection of each individual peptide into 2 animals. As a result, 4 promising new molecular imaging agents were identified that otherwise would not have been selected based solely on in vitro data. This study is the first example of the practical application of a high-throughput screening approach using microPET imaging of [(18)F]-labeled peptides for the rapid in vivo identification of potential new molecular imaging agents. PMID- 19815498 TI - Formation of nanoparticles of blue haze enhanced by anthropogenic pollution. AB - The molecular processes leading to formation of nanoparticles of blue haze over forested areas are highly complex and not fully understood. We show that the interaction between biogenic organic acids and sulfuric acid enhances nucleation and initial growth of those nanoparticles. With one cis-pinonic acid and three to five sulfuric acid molecules in the critical nucleus, the hydrophobic organic acid part enhances the stability and growth on the hydrophilic sulfuric acid counterpart. Dimers or heterodimers of biogenic organic acids alone are unfavorable for new particle formation and growth because of their hydrophobicity. Condensation of low-volatility organic acids is hindered on nano sized particles, whereas ammonia contributes negligibly to particle growth in the size range of 3-30 nm. The results suggest that initial growth from the critical nucleus to the detectable size of 2-3 nm most likely occurs by condensation of sulfuric acid and water, implying that anthropogenic sulfur emissions (mainly from power plants) strongly influence formation of terrestrial biogenic particles and exert larger direct and indirect climate forcing than previously recognized. PMID- 19815499 TI - The interaction of TIGIT with PVR and PVRL2 inhibits human NK cell cytotoxicity. AB - NK cell cytotoxicity is controlled by numerous NK inhibitory and activating receptors. Most of the inhibitory receptors bind MHC class I proteins and are expressed in a variegated fashion. It was recently shown that TIGIT, a new protein expressed by T and NK cells binds to PVR and PVR-like receptors and inhibits T cell activity indirectly through the manipulation of DC activity. Here, we show that TIGIT is expressed by all human NK cells, that it binds PVR and PVRL2 but not PVRL3 and that it inhibits NK cytotoxicity directly through its ITIM. Finally, we show that TIGIT counter inhibits the NK-mediated killing of tumor cells and protects normal cells from NK-mediated cytotoxicity thus providing an "alternative self" mechanism for MHC class I inhibition. PMID- 19815500 TI - Structural evolution of p53, p63, and p73: implication for heterotetramer formation. AB - Oligomerization of members of the p53 family of transcription factors (p53, p63, and p73) is essential for their distinct functions in cell-cycle control and development. To elucidate the molecular basis for tetramer formation of the various family members, we solved the crystal structure of the human p73 tetramerization domain (residues 351-399). Similarly to the canonical p53 tetramer, p73 forms a tetramer with D(2) symmetry that can be described as a dimer of dimers. The most striking difference between the p53 and p73 tetramerization domain is the presence of an additional C-terminal helix in p73. This helix, which is conserved in p63, is essential for stabilizing the overall architecture of the tetramer, as evidenced by the different oligomeric structures observed for a shortened variant lacking this helix. The helices act as clamps, wrapping around the neighboring dimer and holding it in place. In addition, we show by mass spectrometry that the tetramerization domains of p63 and p73, but not p53, fully exchange, with different mixed tetramers present at equilibrium, albeit at a relatively slow rate. Taken together, these data provide intriguing insights into the divergent evolution of the oligomerization domain within the p53 family, from the ancestral p63/p73-like protein toward smaller, less promiscuous monomeric building blocks in human p53, allowing functional separation of the p53 pathway from that of its family members. PMID- 19815501 TI - Mechanisms of NKT cell anergy induction involve Cbl-b-promoted monoubiquitination of CARMA1. AB - Repeated injection of alpha-galactosylceramide, an agonistic ligand for natural killer T (NKT) cells, results in long-term unresponsiveness or anergy, which severely limits its clinical application. However, the molecular mechanisms leading to NKT anergy induction remain unclear. We show here that the decreased IFN-gamma production and failed tumor rejection observed in anergized NKT cells are rescued by Cbl-b deficiency. Cbl-b E3 ligase activity is critical for the anergy induction, as revealed by the similarity between Cbl-b(-/-) and its RING finger mutant NKT cells. Cbl-b binds and promotes monoubiquitination to CARMA1, a critical signaling molecule in NFkappaB activation. Ubiquitin conjugation to CARMA1 disrupts its complex formation with Bcl10 without affecting its protein stability. In addition, CARMA1(-/-) NKT cells are defective in IFN-gamma production. The study identifies an important signaling pathway linking Cbl-b induced monoubiquitination to NFkappaB activation in NKT cell anergy induction, which may help design approaches for human cancer therapy. PMID- 19815502 TI - The crystal structure of sphingosine-1-phosphate in complex with a Fab fragment reveals metal bridging of an antibody and its antigen. AB - The pleiotropic signaling lipid sphingosine-1-phosphate (S1P) plays significant roles in angiogenesis, heart disease, and cancer. LT1009 (also known as sonepcizumab) is a humanized monoclonal antibody that binds S1P with high affinity and specificity. Because the antibody is currently in clinical trials, it is important to confirm by structural and biochemical analyses that it binds its target in a predictable manner. Therefore, we determined the structure of a complex between the LT1009 antibody Fab fragment and S1P refined to 1.90 A resolution. The antibody employs unique and diverse strategies to recognize its antigen. Two metal ions bridge complementarity determining regions from the antibody light chain and S1P. The coordination geometry, inductively coupled plasma spectroscopy, surface plasmon resonance spectroscopy, and biochemical assays suggest that these are Ca(2+). The amino alcohol head group of the sphingosine backbone is recognized through hydrogen bonding interactions from 1 aa side chain and polypeptide backbone atoms of the antibody light and heavy chains. The S1P hydrophobic tail is almost completely enclosed within a hydrophobic channel formed primarily by the heavy chain. Both treatment of the complex with metal chelators and mutation of amino acids in the light chain that coordinate the metal atoms or directly contact the polar head group abrogate binding, while mutations within the hydrophobic cavity also decrease S1P binding affinity. The structure suggests mechanistic details for recognition of a signaling lipid by a therapeutic antibody candidate. Moreover, this study provides direct structural evidence that antibodies are capable of using metals to bridge antigen:antibody complexes. PMID- 19815503 TI - Mechanisms of gold biomineralization in the bacterium Cupriavidus metallidurans. AB - While the role of microorganisms as main drivers of metal mobility and mineral formation under Earth surface conditions is now widely accepted, the formation of secondary gold (Au) is commonly attributed to abiotic processes. Here we report that the biomineralization of Au nanoparticles in the metallophillic bacterium Cupriavidus metallidurans CH34 is the result of Au-regulated gene expression leading to the energy-dependent reductive precipitation of toxic Au(III) complexes. C. metallidurans, which forms biofilms on Au grains, rapidly accumulates Au(III)-complexes from solution. Bulk and microbeam synchrotron X-ray analyses revealed that cellular Au accumulation is coupled to the formation of Au(I)-S complexes. This process promotes Au toxicity and C. metallidurans reacts by inducing oxidative stress and metal resistances gene clusters (including a Au specific operon) to promote cellular defense. As a result, Au detoxification is mediated by a combination of efflux, reduction, and possibly methylation of Au complexes, leading to the formation of Au(I)-C-compounds and nanoparticulate Au(0). Similar particles were observed in bacterial biofilms on Au grains, suggesting that bacteria actively contribute to the formation of Au grains in surface environments. The recognition of specific genetic responses to Au opens the way for the development of bioexploration and bioprocessing tools. PMID- 19815504 TI - Tumor-associated and microbial proteases compromise host IgG effector functions by a single cleavage proximal to the hinge. AB - The successful elimination of pathogenic cells and microorganisms by the humoral immune system relies on effective interactions between host immunoglobulins and Fc gamma receptors on effector cells, in addition to the complement system. Essential Ig motifs that direct those interactions reside within the conserved IgG lower hinge/CH2 interface. We noted that a group of tumor-related and microbial proteases cleaved human IgG1s in that region, and the "nick" of just one of the heavy chains profoundly inhibited IgG1 effector functions. We focused on IgG1 monoclonal antibodies (mAbs) since IgG1 is the most abundant human subclass and demonstrates robust Fc-mediated effector functions. The loss of Fc mediated cell killing activities was correlated with diminished binding to the Fc gamma family of receptors, but a similar decrease in affinity was not observed toward the FcRn receptor that maintains IgG in circulation. Endogenous human IgG cleavage products of comparable size to mAbs with the single cleavage were detected by Western blot analysis in synovial fluid from patients with rheumatoid arthritis and in breast carcinoma extracts. Their detection is problematic under physiological conditions, since there is no loss of structure, and antigen binding capability is unaffected. These findings suggest that within the hostile proteolytic microenvironments associated with many diseases, key effector functions of host IgGs, or therapeutic Abs, may be compromised. PMID- 19815505 TI - Olfactory perceptual learning requires adult neurogenesis. AB - Perceptual learning is required for olfactory function to adapt appropriately to changing odor environments. We here show that newborn neurons in the olfactory bulb are not only involved in, but necessary for, olfactory perceptual learning. First, the discrimination of perceptually similar odorants improves in mice after repeated exposure to the odorants. Second, this improved discrimination is accompanied by an elevated survival rate of newborn inhibitory neurons, preferentially involved in processing of the learned odor, within the olfactory bulb. Finally, blocking neurogenesis before and during the odorant exposure period prevents this learned improvement in discrimination. Olfactory perceptual learning is thus mediated by the reinforcement of functional inhibition in the olfactory bulb by adult neurogenesis. PMID- 19815506 TI - Negative regulation of MyD88-dependent signaling by IL-10 in dendritic cells. AB - IL-10 produced by dendritic cells (DC) can limit or terminate ongoing inflammatory responses by inhibiting the proinflammatory cytokine production. Currently, the molecular mechanism by which IL-10 suppresses cytokine production is still ill-defined. In this study, we showed that IL-10 produced by DC dampens myeloid differentiation factor (MyD)88-dependent, but not MyD88-independent signaling. At the molecular level, IL-10 induces ubiquitination and subsequent protein degradation of MyD88-dependent signaling molecules, including IL-1 receptor-associated kinase 4 and TNF-receptor associated factor 6. Protein degradation by IL-10 was associated with decreased phosphorylation of p38, JNK, and IKK. All of these events were prevented by either blocking IL-10 receptor signaling or inhibiting proteasome degradation. IL-10 induced LPS hyporesponsiveness using the same mechanisms, i.e., ubiquitination and protein degradation. Thus, a previously undescribed regulatory mechanism by which IL-10 mediated protein degradation contributes to the inhibition of inflammatory cytokine production and endotoxin tolerance in DC. PMID- 19815507 TI - Epstein-Barr virus LMP2A bypasses p53 inactivation in a MYC model of lymphomagenesis. AB - Although Epstein-Barr virus (EBV) is linked to Burkitt's lymphoma (BL), the role of the virus in lymphomagenesis is unclear. LMP2A, encoded by EBV, can be detected in BL biopsies and has prosurvival functions. We generated mice expressing MYC and LMP2A in B cells. LMP2A/lambda-MYC mice show greatly accelerated tumor onset. Similar to previous work, we found p53 mutations in lambda-MYC tumors; however, we detected no mutations in the rapidly arising LMP2A/lambda-MYC tumors. We further demonstrate that the p53 pathway is functionally intact in LMP2A/lambda-MYC tumors, which have increased levels of PUMA and sensitivity to p53 activation by Nutlin. This work shows that LMP2A can permit tumorigenesis in the presence of an intact p53 pathway, identifying an important contribution of EBV to BL. PMID- 19815508 TI - Macroevolution and the biological diversity of plants and herbivores. AB - Terrestrial biodiversity is dominated by plants and the herbivores that consume them, and they are one of the major conduits of energy flow up to higher trophic levels. Here, we address the processes that have generated the spectacular diversity of flowering plants (>300,000 species) and insect herbivores (likely >1 million species). Long-standing macroevolutionary hypotheses have postulated that reciprocal evolution of adaptations and subsequent bursts of speciation have given rise to much of this biodiversity. We critically evaluate various predictions based on this coevolutionary theory. Phylogenetic reconstruction of ancestral states has revealed evidence for escalation in the potency or variety of plant lineages' chemical defenses; however, escalation of defense has been moderated by tradeoffs and alternative strategies (e.g., tolerance or defense by biotic agents). There is still surprisingly scant evidence that novel defense traits reduce herbivory and that such evolutionary novelty spurs diversification. Consistent with the coevolutionary hypothesis, there is some evidence that diversification of herbivores has lagged behind, but has nevertheless been temporally correlated with that of their host-plant clades, indicating colonization and radiation of insects on diversifying plants. However, there is still limited support for the role of host-plant shifts in insect diversification. Finally, a frontier area of research, and a general conclusion of our review, is that community ecology and the long-term evolutionary history of plant and insect diversification are inexorably intertwined. PMID- 19815509 TI - Miz1 is a signal- and pathway-specific modulator or regulator (SMOR) that suppresses TNF-alpha-induced JNK1 activation. AB - The proinflammatory cytokine TNF-alpha exerts its pleiotropic functions through activation of multiple downstream effectors, including JNK1. Yet, the underlying regulatory mechanism is incompletely understood. Here, we report that the transcription factor Myc-interacting zinc-finger protein 1 (Miz1) selectively suppresses TNF-alpha-induced JNK1 activation and cell death independently of its transcription activity. Proteomics analysis and yeast two-hybrid screening reveal that Miz1 is a JNK-associated protein. The TNF-alpha-induced activation of JNK1 is augmented in Miz1-deficient mouse embryonic fibroblasts (Miz1(-/-) MEFs), but the augmentation is abrogated by reintroduction of Miz1 or its transcription deficient mutant. The regulation by Miz1 is highly specific, because it regulates TNF-alpha-induced TRAF2 K63-linked polyubiquitination. Neither JNK1 activation by IL-1beta or UV nor TNF-alpha-induced activation of p38, ERK, or IkappaB kinase complex is affected by the loss of Miz1. The TNF-alpha-induced cell death also is accelerated in Miz1(-/-) MEFs. Upon TNF-alpha stimulation, Miz1 is degraded rapidly by the proteasome, relieving its suppression on JNK1 activation. Thus, our results show that in addition to being a transcription factor Miz1 acts as a signal- and pathway-specific modulator or regulator that specifically regulates TNF-alpha-induced JNK1 activation and cell death. PMID- 19815510 TI - Beta-adrenergic signaling accelerates and synchronizes cardiac ryanodine receptor response to a single L-type Ca2+ channel. AB - As the most prototypical G protein-coupled receptor, beta-adrenergic receptor (betaAR) regulates the pace and strength of heart beating by enhancing and synchronizing L-type channel (LCC) Ca(2+) influx, which in turn elicits greater sarcoplasmic reticulum (SR) Ca(2+) release flux via ryanodine receptors (RyRs). However, whether and how betaAR-protein kinase A (PKA) signaling directly modulates RyR function remains elusive and highly controversial. By using unique single-channel Ca(2+) imaging technology, we measured the response of a single RyR Ca(2+) release unit, in the form of a Ca(2+) spark, to its native trigger, the Ca(2+) sparklet from a single LCC. We found that acute application of the selective betaAR agonist isoproterenol (1 microM, < or = 20 min) increased triggered spark amplitude in an LCC unitary current-independent manner. The increased ratio of Ca(2+) release flux underlying a Ca(2+) spark to SR Ca(2+) content indicated that betaAR stimulation helps to recruit additional RyRs in synchrony. Quantification of sparklet-spark kinetics showed that betaAR stimulation synchronized the stochastic latency and increased the fidelity (i.e., chance of hit) of LCC-RyR intermolecular signaling. The RyR modulation was independent of the increased SR Ca(2+) content. The PKA antagonists Rp-8-CPT-cAMP (100 microM) and H89 (10 microM) both eliminated these effects, indicating that betaAR acutely modulates RyR activation via the PKA pathway. These results demonstrate unequivocally that RyR activation by a single LCC is accelerated and synchronized during betaAR stimulation. This molecular mechanism of sympathetic regulation will permit more fundamental studies of altered betaAR effects in cardiovascular diseases. PMID- 19815511 TI - Rapid DNA loss as a counterbalance to genome expansion through retrotransposon proliferation in plants. AB - Transposable elements, particularly LTR-retrotransposons, comprise the primary vehicle for genome size expansion in plants, while DNA removal through illegitimate recombination and intrastrand homologous recombination serve as the most important counteracting forces to plant genomic obesity. Despite extensive research, the relative impact of these opposing forces and hence the directionality of genome size change remains unknown. In Gossypium (cotton), the 3-fold genome size variation among diploids is due largely to copy number variation of the gypsy-like retrotransposon Gorge3. Here we combine comparative sequence analysis with a modeling approach to study the directionality of genome size change in Gossypium. We demonstrate that the rate of DNA removal in the smaller genomes is sufficient to reverse genome expansion through Gorge3 proliferation. These data indicate that rates of DNA loss can be highly variable even within a single plant genus, and that the known mechanisms of DNA loss can indeed reverse the march toward genomic obesity. PMID- 19815512 TI - Theoretical examination of quantum coherence in a photosynthetic system at physiological temperature. AB - The observation of long-lived electronic coherence in a photosynthetic pigment protein complex, the Fenna-Matthews-Olson (FMO) complex, is suggestive that quantum coherence might play a significant role in achieving the remarkable efficiency of photosynthetic electronic energy transfer (EET), although the data were acquired at cryogenic temperature [Engel GS, et al. (2007) Evidence for wavelike energy transfer through quantum coherence in photosynthetic systems. Nature 446:782-786]. In this paper, the spatial and temporal dynamics of EET through the FMO complex at physiological temperature are investigated theoretically. The numerical results reveal that quantum wave-like motion persists for several hundred femtoseconds even at physiological temperature, and suggest that the FMO complex may work as a rectifier for unidirectional energy flow from the peripheral light-harvesting antenna to the reaction center complex by taking advantage of quantum coherence and the energy landscape of pigments tuned by the protein scaffold. A potential role of quantum coherence is to overcome local energetic traps and aid efficient trapping of electronic energy by the pigments facing the reaction center complex. PMID- 19815513 TI - Peroxisome dependency of alkyl-containing GPI-anchor biosynthesis in the endoplasmic reticulum. AB - Glycosylphosphatidylinositol-anchored proteins (GPI-APs) play various roles in cell-cell and cell-environment interactions. GPI is synthesized in the endoplasmic reticulum (ER) from phosphatidylinositol (PI) through step-wise reactions including transfers of monosaccharides and preassembled GPI is transferred en bloc to proteins. Cellular PI contains mostly diacyl glycerol and unsaturated fatty acid in the sn-2 position, whereas mammalian GPI-APs have mainly 1-alkyl-2-acyl PI and almost exclusively stearic acid, a saturated chain, at the sn-2 position. The latter characteristic is the result of fatty acid remodeling occurring in the Golgi, generating GPI-anchors compatible with raft membrane. The former characteristic is the result of diacyl to alkyl-acyl change occurring in the third GPI intermediate, glucosaminyl-inositolacylated-PI (GlcN acyl-PI). Here we investigated the origin of the sn-1 alkyl-chain in GPI-APs. Using cell lines defective in the peroxisomal alkyl-phospholipid biosynthetic pathway, we demonstrated that generation of alkyl-containing GPI is dependent upon the peroxisomal pathway. We further demonstrated that in cells defective in the peroxisome pathway, the chain composition of the diacyl glycerol moiety in GlcN-acyl-PI is different from those in the first intermediate N acetylglucosaminyl-PI and cellular PI, indicating that not only diacyl to alkyl acyl change but also diacyl to diacyl change occurs in GlcN-acyl-PI. We therefore propose a biosynthetic step within GlcN-acyl-PI in which the diacyl glycerol (or diacyl phosphatidic acid) part is replaced by diradyl glycerol (or diradyl phosphatidic acid). These results highlight cooperation of three organelles, the ER, the Golgi, and the peroxisome, in the generation of the lipid portion of GPI APs. PMID- 19815514 TI - 2D IR provides evidence for mobile water molecules in beta-amyloid fibrils. AB - The motion of water molecules close to amide groups causes their vibrational frequencies to vary rapidly in time. These variations are uniquely sensed by 2 dimensional infrared spectroscopy (2D IR). Here, it is proposed from 2 dimensional experiments on fibrils of amyloid beta (Abeta)40 that there are water molecules in the fibrils. The spatial locations of the water (D(2)O) were inferred from the responses of 18 amide modes of Abeta40 labeled with (13)C = (18)O. Fast frequency variations were found for residues L17 and V18 and for the apposed residues L34 and V36, suggesting cavities or channels containing mobile water molecules can form between the 2 sheets. Spectroscopic analysis showed that there are 1.2 water molecules per strand in the fibrils. The (13)C = (18)O substitution of 1 residue per strand creates a linear array of isotopologs along the fibril axis that manifests clearly identifiable vibrational transitions. Here, it is shown from the distributions of amide-I' vibrational frequencies that the regularity of these chains is strongly residue dependent and in most cases the distorted regions are also those associated with the putative mobile water molecules. It is proposed that Abeta40 fibrils contain structurally significant mobile water molecules within the intersheet region. PMID- 19815515 TI - Water-mediated signal multiplication with Y-shaped carbon nanotubes. AB - Molecular scale signal conversion and multiplication is of particular importance in many physical and biological applications, such as molecular switches, nano gates, biosensors, and various neural systems. Unfortunately, little is currently known regarding the signal processing at the molecular level, partly due to the significant noises arising from the thermal fluctuations and interferences between branch signals. Here, we use molecular dynamics simulations to show that a signal at the single-electron level can be converted and multiplied into 2 or more signals by water chains confined in a narrow Y-shaped nanochannel. This remarkable transduction capability of molecular signal by Y-shaped nanochannel is found to be attributable to the surprisingly strong dipole-induced ordering of such water chains, such that the concerted water orientations in the 2 branches of the Y-shaped nanotubes can be modulated by the water orientation in the main channel. The response to the switching of the charge signal is very rapid, from a few nanoseconds to a few hundred nanoseconds. Furthermore, simulations with various water models, including TIP3P, TIP4P, and SPC/E, show that the transduction capability of the Y-shaped carbon nanotubes is very robust at room temperature, with the interference between branch signals negligible. PMID- 19815516 TI - Age-associated increase in lifespan of naive CD4 T cells contributes to T-cell homeostasis but facilitates development of functional defects. AB - With age, T-cell generation from the thymus is much reduced, yet a substantial naive T-cell pool is maintained even in aged animals, suggesting that naive T cells either persist longer or turn over faster to maintain T-cell homeostasis. We found that with age, naive CD4 T cells became progressively longer-lived. Their longer lifespan did not depend on recognition of self-peptide/class II. Newly generated naive T cells derived from aged stem cells had a shorter lifespan, like that of young naive T cells. Conversely, naive CD4 T cells derived from middle-aged thymectomized mice were longer-lived in vivo, and their development of functional defects was accelerated. These observations suggest that naive T cells develop their longer lifespan during their sojourn in the periphery. Increased longevity of naive CD4 T cells correlated well with reduced expression of proapoptotic molecule Bim. We suggest that the intrinsic increase in longevity helps maintain naive T-cell homeostasis but facilitates the development of functional defects in mice. PMID- 19815517 TI - Uncovering pathways in DNA oligonucleotide hybridization via transition state analysis. AB - DNA hybridization plays a central role in biology and, increasingly, in materials science. Yet, there is no precedent for examining the pathways by which specific single-stranded DNA sequences interact to assemble into a double helix. A detailed model of DNA is adopted in this work to examine such pathways and to determine the role of sequence, if any, on DNA hybridization. Transition path sampling simulations reveal that DNA rehybridization is prompted by a distinct nucleation event involving molecular sites with approximately four bases pairing with partners slightly offset from those involved in ideal duplexation. Nucleation is promoted in regions with repetitive base pair sequence motifs, which yield multiple possibilities for finding complementary base partners. Repetitive sequences follow a nonspecific pathway to renaturation consistent with a molecular "slithering" mechanism, whereas random sequences favor a restrictive pathway involving the formation of key base pairs before renaturation fully ensues. PMID- 19815518 TI - Feed-forward inhibition as a buffer of the neuronal input-output relation. AB - Neuronal processing depends on the input-output (I/O) relation between the frequency of synaptic stimulation and the resultant axonal firing rate. The all or-none properties of spike generation and active membrane mechanisms can make the neuronal I/O relation very steep. The ensuing nearly bimodal behavior may severely limit information coding, as minimal input fluctuations within the expected natural variability could cause neuronal output to jump between quiescence and maximum firing rate. Here, using biophysically and anatomically realistic computational models of individual neurons, we demonstrate that feed forward inhibition, a ubiquitous mechanism in which inhibitory interneurons and their target cells are activated by the same excitatory input, can change a steeply sigmoid I/O curve into a double-sigmoid typical of buffer systems. The addition of an intermediate plateau stabilizes the spiking response over a broad dynamic range of input frequency, ensuring robust integration of noisy synaptic signals. Both the buffered firing rate and its input firing range can be independently and extensively modulated by biologically plausible changes in the weight and number of excitatory synapses on the feed-forward interneuron. By providing a soft switch between essentially digital and analog rate-code, this continuous control of the circuit I/O could dramatically increase the computational power of neuronal integration. PMID- 19815519 TI - Temporomandibular joint formation requires two distinct hedgehog-dependent steps. AB - We conducted a genetic analysis of the developing temporo-mandibular or temporomandi-bular joint (TMJ), a highly specialized synovial joint that permits movement and function of the mammalian jaw. First, we used laser capture microdissection to perform a genome-wide expression analysis of each of its developing components. The expression patterns of genes identified in this screen were examined in the TMJ and compared with those of other synovial joints, including the shoulder and the hip joints. Striking differences were noted, indicating that the TMJ forms via a distinct molecular program. Several components of the hedgehog (Hh) signaling pathway are among the genes identified in the screen, including Gli2, which is expressed specifically in the condyle and in the disk of the developing TMJ. We found that mice deficient in Gli2 display aberrant TMJ development such that the condyle loses its growth-plate-like cellular organization and no disk is formed. In addition, we used a conditional strategy to remove Smo, a positive effector of the Hh signaling pathway, from chondrocyte progenitors. This cell autonomous loss of Hh signaling allows for disk formation, but the resulting structure fails to separate from the condyle. Thus, these experiments establish that Hh signaling acts at two distinct steps in disk morphogenesis, condyle initiation, and disk-condyle separation and provide a molecular framework for future studies of the TMJ. PMID- 19815520 TI - Alpha-actinin2 cytoskeletal protein is required for the functional membrane localization of a Ca2+-activated K+ channel (SK2 channel). AB - The importance of proper ion channel trafficking is underpinned by a number of channel-linked genetic diseases whose defect is associated with failure to reach the cell surface. Conceptually, it is reasonable to suggest that the function of ion channels depends critically on the precise subcellular localization and the number of channel proteins on the cell surface membrane, which is determined jointly by the secretory and endocytic pathways. Yet the precise mechanisms of the entire ion channel trafficking pathway remain unknown. Here, we directly demonstrate that proper membrane localization of a small-conductance Ca(2+) activated K(+) channel (SK2 or K(Ca)2.2) is dependent on its interacting protein, alpha-actinin2, a major F-actin crosslinking protein. SK2 channel localization on the cell-surface membrane is dynamically regulated, and one of the critical steps includes the process of cytoskeletal anchoring of SK2 channel by its interacting protein, alpha-actinin2, as well as endocytic recycling via early endosome back to the cell membrane. Consequently, alteration of these components of SK2 channel recycling results in profound changes in channel surface expression. The importance of our findings may transcend the area of K(+) channels, given that similar cytoskeletal interaction and anchoring may be critical for the membrane localization of other ion channels in neurons and other excitable cells. PMID- 19815521 TI - Some quantum weirdness in physiology. PMID- 19815522 TI - Trade-offs and synergies between carbon storage and livelihood benefits from forest commons. AB - Forests provide multiple benefits at local to global scales. These include the global public good of carbon sequestration and local and national level contributions to livelihoods for more than half a billion users. Forest commons are a particularly important class of forests generating these multiple benefits. Institutional arrangements to govern forest commons are believed to substantially influence carbon storage and livelihood contributions, especially when they incorporate local knowledge and decentralized decision making. However, hypothesized relationships between institutional factors and multiple benefits have never been tested on data from multiple countries. By using original data on 80 forest commons in 10 countries across Asia, Africa, and Latin America, we show that larger forest size and greater rule-making autonomy at the local level are associated with high carbon storage and livelihood benefits; differences in ownership of forest commons are associated with trade-offs between livelihood benefits and carbon storage. We argue that local communities restrict their consumption of forest products when they own forest commons, thereby increasing carbon storage. In showing rule-making autonomy and ownership as distinct and important institutional influences on forest outcomes, our results are directly relevant to international climate change mitigation initiatives such as Reduced Emissions from Deforestation and Forest Degradation (REDD) and avoided deforestation. Transfer of ownership over larger forest commons patches to local communities, coupled with payments for improved carbon storage can contribute to climate change mitigation without adversely affecting local livelihoods. PMID- 19815523 TI - Functional interchangeability of rod and cone transducin alpha-subunits. AB - Rod and cone photoreceptors use similar but distinct sets of phototransduction proteins to achieve different functional properties, suitable for their role as dim and bright light receptors, respectively. For example, rod and cone visual pigments couple to distinct variants of the heterotrimeric G protein transducin. However, the role of the structural differences between rod and cone transducin alpha subunits (Talpha) in determining the functional differences between rods and cones is unknown. To address this question, we studied the translocation and signaling properties of rod Talpha expressed in cones and cone Talpha expressed in rods in three mouse strains: rod Talpha knockout, cone Talpha GNAT2(cpfl3) mutant, and rod and cone Talpha double mutant rd17 mouse. Surprisingly, although the rod/cone Talpha are only 79% identical, exogenously expressed rod or cone Talpha localized and translocated identically to endogenous Talpha in each photoreceptor type. Moreover, exogenously expressed rod or cone Talpha rescued electroretinogram responses (ERGs) in mice lacking functional cone or rod Talpha, respectively. Ex vivo transretinal ERG and single-cell recordings from rd17 retinas treated with rod or cone Talpha showed comparable rod sensitivity and response kinetics. These results demonstrate that cone Talpha forms a functional heterotrimeric G protein complex in rods and that rod and cone Talpha couple equally well to the rod phototransduction cascade. Thus, rod and cone transducin alpha-subunits are functionally interchangeable and their signaling properties do not contribute to the intrinsic light sensitivity differences between rods and cones. Additionally, the technology used here could be adapted for any such homologue swap desired. PMID- 19815524 TI - Substrate positioning controls the partition between halogenation and hydroxylation in the aliphatic halogenase, SyrB2. AB - The alpha-ketoglutarate-dependent hydroxylases and halogenases employ similar reaction mechanisms involving hydrogen-abstracting Fe(IV)-oxo (ferryl) intermediates. In the halogenases, the carboxylate residue from the His(2)(Asp/Glu)(1) "facial triad" of iron ligands found in the hydroxylases is replaced by alanine, and a halide ion (X(-)) coordinates at the vacated site. Halogenation is thought to result from "rebound" of the halogen radical from the X-Fe(III)-OH intermediate produced by hydrogen (H(*)) abstraction to the substrate radical. The alternative decay pathway for the X-Fe(III)-OH intermediate, rebound of the hydroxyl radical to the substrate radical (as occurs in the hydroxylases), reportedly does not compete. Here we show for the halogenase SyrB2 that positioning of the alkyl group of the substrate away from the oxo/hydroxo ligand and closer to the halogen ligand sacrifices H(*) abstraction proficiency for halogen-rebound selectivity. Upon replacement of L Thr, the C4 amino acid tethered to the SyrB1 carrier protein in the native substrate, by the C5 amino acid L-norvaline, decay of the chloroferryl intermediate becomes 130x faster and the reaction outcome switches to primarily hydroxylation of C5, consistent with projection of the methyl group closer to the oxo/hydroxo by the longer side chain. Competing H(*) abstraction from C4 results primarily in chlorination, as occurs at this site in the native substrate. Consequently, deuteration of C5, which slows attack at this site, switches both the regioselectivity from C5 to C4 and the chemoselectivity from hydroxylation to chlorination. Thus, substrate-intermediate disposition and the carboxylate --> halide ligand swap combine to specify the halogenation outcome. PMID- 19815525 TI - Comparative genomics reveal the mechanism of the parallel evolution of O157 and non-O157 enterohemorrhagic Escherichia coli. AB - Among the various pathogenic Escherichia coli strains, enterohemorrhagic E. coli (EHEC) is the most devastating. Although serotype O157:H7 strains are the most prevalent, strains of different serotypes also possess similar pathogenic potential. Here, we present the results of a genomic comparison between EHECs of serotype O157, O26, O111, and O103, as well as 21 other, fully sequenced E. coli/Shigella strains. All EHECs have much larger genomes (5.5-5.9 Mb) than the other strains and contain surprisingly large numbers of prophages and integrative elements (IEs). The gene contents of the 4 EHECs do not follow the phylogenetic relationships of the strains, and they share virulence genes for Shiga toxins and many other factors. We found many lambdoid phages, IEs, and virulence plasmids that carry the same or similar virulence genes but have distinct evolutionary histories, indicating that independent acquisition of these mobile genetic elements has driven the evolution of each EHEC. Particularly interesting is the evolution of the type III secretion system (T3SS). We found that the T3SS of EHECs is composed of genes that were introduced by 3 different types of genetic elements: an IE referred to as the locus of enterocyte effacement, which encodes a central part of the T3SS; SpLE3-like IEs; and lambdoid phages carrying numerous T3SS effector genes and other T3SS-related genes. Our data demonstrate how E. coli strains of different phylogenies can independently evolve into EHECs, providing unique insights into the mechanisms underlying the parallel evolution of complex virulence systems in bacteria. PMID- 19815526 TI - Distinct and overlapping roles for two Dicer-like proteins in the RNA interference pathways of the ancient eukaryote Trypanosoma brucei. AB - Trypanosoma brucei is one of the most ancient eukaryotes where RNA interference (RNAi) is operational and is the only single-cell pathogen where RNAi has been extensively studied and used as a tool for functional analyses. Here, we report that the T. brucei RNAi pathway, although relying on a single Argonaute protein (AGO1), is initiated by the activities of two distinct Dicer-like enzymes. Both TbDCL1, a mostly cytoplasmic protein, and the previously undescribed nuclear enzyme TbDCL2 contribute to the biogenesis of siRNAs from retroposons. However, TbDCL2 has a predominant role in generating siRNAs from chromosomal internal repeat transcripts that accumulate at the nucleolus in RNAi-deficient cells and in initiating the endogenous RNAi response against retroposons and repeats alike. Moreover, siRNAs generated by both TbDCL1 and TbDCL2 carry a 5'-monophosphate and a blocked 3' terminus, suggesting that 3' end modification is an ancient trait of siRNAs. We thus propose a model whereby TbDCL2 fuels the T. brucei nuclear RNAi pathway and TbDCL1 patrols the cytoplasm, posttranscriptionally silencing potentially harmful nucleic acid parasites that may access the cytoplasm. Nevertheless, we also provide evidence for cross-talk between the two Dicer-like enzymes, because TbDCL2 is implicated in the generation of 35- to 65-nucleotide intermediate transcripts that appear to be substrates for TbDCL1. Our finding that dcl2KO cells are more sensitive to RNAi triggers than wild-type cells has significant implications for reverse genetic analyses in this important human pathogen. PMID- 19815527 TI - Structural imperatives impose diverse evolutionary constraints on helical membrane proteins. AB - The amino acid sequences of transmembrane regions of helical membrane proteins are highly constrained, diverging at slower rates than their extramembrane regions and than water-soluble proteins. Moreover, helical membrane proteins seem to fall into fewer families than water-soluble proteins. The reason for the differential restrictions on sequence remains unexplained. Here, we show that the evolution of transmembrane regions is slowed by a previously unrecognized structural constraint: Transmembrane regions bury more residues than extramembrane regions and soluble proteins. This fundamental feature of membrane protein structure is an important contributor to the differences in evolutionary rate and to an increased susceptibility of the transmembrane regions to disease causing single-nucleotide polymorphisms. PMID- 19815528 TI - Simultaneous imaging of tumor oxygenation and microvascular permeability using Overhauser enhanced MRI. AB - Architectural and functional abnormalities of blood vessels are a common feature in tumors. A consequence of increased vascular permeability and concomitant aberrant blood flow is poor delivery of oxygen and drugs, which is associated with treatment resistance. In the present study, we describe a strategy to simultaneously visualize tissue oxygen concentration and microvascular permeability by using a hyperpolarized (1)H-MRI, known as Overhauser enhanced MRI (OMRI), and an oxygen-sensitive contrast agent OX63. Substantial MRI signal enhancement was induced by dynamic nuclear polarization (DNP). The DNP achieved up to a 7,000% increase in MRI signal at an OX63 concentration of 1.5 mM compared with that under thermal equilibrium state. The extent of hyperpolarization is influenced mainly by the local concentration of OX63 and inversely by the tissue oxygen level. By collecting dynamic OMRI images at different hyperpolarization levels, local oxygen concentration and microvascular permeability of OX63 can be simultaneously determined. Application of this modality to murine tumors revealed that tumor regions with high vascular permeability were spatio-temporally coincident with hypoxia. Quantitative analysis of image data from individual animals showed an inverse correlation between tumor vascular leakage and median oxygen concentration. Immunohistochemical analyses of tumor tissues obtained from the same animals after OMRI experiments demonstrated that lack of integrity in tumor blood vessels was associated with increased tumor microvascular permeability. This dual imaging technique may be useful for the longitudinal assessment of changes in tumor vascular function and oxygenation in response to chemotherapy, radiotherapy, or antiangiogenic treatment. PMID- 19815529 TI - AMP-activated protein kinase adapts rRNA synthesis to cellular energy supply. AB - AMP-activated protein kinase (AMPK) senses changes in the intracellular AMP/ATP ratio, switching off energy-consuming processes and switching on catabolic pathways in response to energy depletion. Here, we show that AMPK down-regulates rRNA synthesis under glucose restriction by phosphorylating the RNA polymerase I (Pol I)-associated transcription factor TIF-IA at a single serine residue (Ser 635). Phosphorylation by AMPK impairs the interaction of TIF-IA with the TBP containing promoter selectivity factor SL1, thereby precluding the assembly of functional transcription initiation complexes. Mutation of Ser-635 compromises down-regulation of Pol I transcription in response to low energy supply, supporting that activation of AMPK adapts rRNA synthesis to nutrient availability and the cellular energy status. PMID- 19815530 TI - CD4+ T cells control the differentiation of Gr1+ monocytes into fibrocytes. AB - Fibrocytes are collagen-type-I-producing cells that arise at low frequency from hematopoietic cells. We have analyzed in mice which leukocyte subsets are required for generation of fibrocytes and show that murine fibrocytes develop from the subpopulation of CD11b(+) CD115(+) Gr1(+) monocytes under the control of CD4(+) T cells. In the absence of CD4(+) T cells, differentiation of fibrocytes was markedly reduced in vitro and in vivo. In the presence of CD4(+) T cells, the characteristics of T-cell activation critically determined development of fibrocytes. Polyclonal activation of CD4(+) T cells induced the release of soluble factors that completely prevented the outgrowth of fibrocytes and could be identified as IL-2, TNF, IFN-gamma, and IL-4. Application of IL-2 and TNF significantly reduced the appearance of fibrocytes and the severity of fibrosis in the model of unilateral ureteral obstruction. In contrast, activation of CD4(+) T cells in the presence of calcineurin inhibitors, but not mTOR inhibitors, markedly enhanced the outgrowth of fibrocytes and renal deposition of collagen I. Taken together, we show that differentiation of fibrocytes is critically dependent on CD4(+) T cells and that the context of T-cell activation determines whether development of fibrocytes is supported or blocked. Our data may have implications for prevention of organ fibrosis in autoimmune diseases and transplantation. PMID- 19815531 TI - Ubiquitin-dependent lysosomal targeting of GABA(A) receptors regulates neuronal inhibition. AB - The strength of synaptic inhibition depends partly on the number of GABA(A) receptors (GABA(A)Rs) found at synaptic sites. The trafficking of GABA(A)Rs within the endocytic pathway is a key determinant of surface GABA(A)R number and is altered in neuropathologies, such as cerebral ischemia. However, the molecular mechanisms and signaling pathways that regulate this trafficking are poorly understood. Here, we report the subunit specific lysosomal targeting of synaptic GABA(A)Rs. We demonstrate that the targeting of synaptic GABA(A)Rs into the degradation pathway is facilitated by ubiquitination of a motif within the intracellular domain of the gamma2 subunit. Blockade of lysosomal activity or disruption of the trafficking of ubiquitinated cargo to lysosomes specifically increases the efficacy of synaptic inhibition without altering excitatory currents. Moreover, mutation of the ubiquitination site within the gamma2 subunit retards the lysosomal targeting of GABA(A)Rs and is sufficient to block the loss of synaptic GABA(A)Rs after anoxic insult. Together, our results establish a previously unknown mechanism for influencing inhibitory transmission under normal and pathological conditions. PMID- 19815532 TI - Promoting oligodendrogenesis and myelin repair using the multiple sclerosis medication glatiramer acetate. AB - The formation of oligodendrocytes (oligodendrogenesis) and myelin is regulated by several neurotrophic factors. Strategies to increase the level of these trophic molecules may facilitate repair in demyelinating conditions, such as multiple sclerosis (MS). Because leukocytes are a source of neurotrophic factors, and as glatiramer acetate (GA) generates T helper 2 (Th2) lymphocytes that are not known to be harmful, we tested the hypothesis that GA regulates oligodendrogenesis and myelin formation. First, we generated GA-reactive Th2 cells and determined that they produced transcripts for neurotrophic factors, including insulin-like growth factor-1 (IGF-1). The conditioned medium from GA-reactive T cells elevated IGF-1 protein and promoted the formation of oligodendrocyte precursor cells (OPCs) from embryonic brain-derived forebrain cells in culture. We next subjected mice to lysolecithin-induced demyelination of the spinal cord. At 7 days after the insult, the number of OPCs in the demyelinated dorsal column was higher than that in uninjured controls, and was further increased by the daily s.c. injection with GA. Increased OPC generation by GA was associated temporally with the elevation of IGF-1 and brain-derived neurotrophic factor (BDNF) in the spinal cord. Finally, the resultant remyelination at 28 days was higher in mice treated with GA during the first 7 days of injury compared with vehicle controls. These results indicate that GA promotes oligodendrogenesis and remyelination through mechanisms that involve the elevation of growth factors conducive for repair. PMID- 19815533 TI - Identification and characterization of the tungsten-containing class of benzoyl coenzyme A reductases. AB - Aromatic compounds are widely distributed in nature and can only be biomineralized by microorganisms. In anaerobic bacteria, benzoyl-CoA (BCoA) is a central intermediate of aromatic degradation, and serves as substrate for dearomatizing BCoA reductases (BCRs). In facultative anaerobes, the mechanistically difficult reduction of BCoA to cyclohexa-1,5-dienoyl-1-carboxyl CoA (dienoyl-CoA) is driven by a stoichiometric ATP hydrolysis, catalyzed by a soluble, three [4Fe-4S] cluster-containing BCR. In this work, an in vitro assay for BCR from the obligately anaerobic Geobacter metallireducens was established. It followed the reverse reaction, the formation of BCoA from dienoyl-CoA in the presence of various electron acceptors. The benzoate-induced activity was highly specific for dienoyl-CoA (K(m) = 24 +/- 4 microM). The corresponding oxygen sensitive enzyme was purified by several chromatographic steps with a 115-fold enrichment and a yield of 18%. The 185-kDa enzyme comprised 73- and 20-kDa subunits, suggesting an alpha(2)beta(2)-composition. MS analysis revealed the subunits as products of the benzoate-induced bamBC genes. The alphabeta unit contained 0.9 W, 15 Fe, and 12.5 acid-labile sulfur. Results from EPR spectroscopy suggest the presence of one [3Fe-4S](0/+1) and three [4Fe-4S](+1/+2) clusters per alphabeta unit; oxidized BamBC exhibited an EPR signal typical for a W(V) species. The FeS clusters and the W- cofactor could only be fully reduced by dienoyl-CoA. BamBC represents the prototype of a previously undescribed class of dearomatizing BCRs that differ completely from the ATP-dependent enzymes from facultative anaerobes. PMID- 19815534 TI - Automated microscopy, dipsticks and the diagnosis of urinary tract infection. AB - OBJECTIVES: Automated microscopy is increasingly used to screen urine samples for suspected urinary tract infection (UTI). A 98.8% negative predictive value has been reported in adult studies. The aim of our study was to validate this method in a paediatric population. METHODS: Urine samples were collected from children with known or suspected nephrourological disease attending nephrology and urology clinics over a 6-week period. Samples were tested with dipstick, the UF-100 flow cytometer (automated microscopy) and culture. A gold standard of a positive culture of morethan 10(5) colony forming units per ml (cfu/ml) with a pathogenic organism was used and the sensitivity, specificity and likelihood ratios were calculated. RESULTS: 280 urine samples were collected from 263 patients (143 male, median age 10.2 years, range 0.1-19.75 years). 221 (79%) were midstream or clean-catch samples. Automated microscopy identified 42 of 186 samples as requiring culture and 17 of 19 samples which had a pure growth of more than 10(5) cfu/ml. Two patients were not identified by automated microscopy: one was treated for vulvovaginitis, and one commenced prophylactic antibiotics prior to the culture result being obtained. The sensitivity, specificity, positive and negative likelihood ratios were 0.89, 0.85, 5.98 and 0.17, respectively. This compared to 0.95, 0.72, 3.34 and 0.29, respectively, with urine dipstick. CONCLUSION: Automated microscopy performed comparably to urine dipstick in the diagnosis of UTI with improved specificity and likelihood ratios with slightly reduced sensitivity. The data support the use of automated microscopy for screening urine samples for culture in children, but different automated microscopy methods and algorithms require local evaluation. PMID- 19815537 TI - Pfetin as a prognostic biomarker in gastrointestinal stromal tumor: novel monoclonal antibody and external validation study in multiple clinical facilities. AB - OBJECTIVE: The clinical course of gastrointestinal stromal tumor (GIST) spans a wide spectrum from a curable disorder to a highly malignant disease that leads to metastasis and death. To develop prognostic modalities for GIST patients, we developed a mouse monoclonal antibody against pfetin, the prognostic value of which has been previously reported. METHODS: The reactivity of the monoclonal antibody against pfetin was examined by western blotting and immunohistochemistry. RESULTS: Western blotting demonstrated that the monoclonal antibody was specific to pfetin. The immunohistochemical study demonstrated that the 5-year disease-free survival rate was 93.2% and 94.5% for GIST patients with pfetin-positive tumors and 70.0% and 80.7% for those with pfetin-negative tumors in the 159 cases from the National Cancer Center Hospital (P < 0.0001) and in the 100 cases from Niigata University Medical and Dental Hospital (P < 0.0001), respectively. Uni- and multivariate analyses revealed that pfetin expression was a powerful prognostic factor among the clinico-pathological parameters examined. CONCLUSIONS: These results establish pfetin as a practical prognostic marker for GIST patients after surgery. Pfetin may also present a novel therapeutic target to prevent recurrence of GIST. PMID- 19815536 TI - The Mycobacterium bovis bacille Calmette-Guerin phagosome proteome. AB - Mycobacterium tuberculosis and Mycobacterium bovis bacille Calmette-Guerin (BCG) alter the maturation of their phagosomes and reside within a compartment that resists acidification and fusion with lysosomes. To define the molecular composition of this compartment, we developed a novel method for obtaining highly purified phagosomes from BCG-infected human macrophages and analyzed the phagosomes by Western immunoblotting and mass spectrometry-based proteomics. Our purification procedure revealed that BCG grown on artificial medium becomes less dense after growth in macrophages. By Western immunoblotting, LAMP-2, Niemann Pick protein C1, and syntaxin 3 were readily detectable on the BCG phagosome but at levels that were lower than on the latex bead phagosome; flotillin-1 and the vacuolar ATPase were barely detectable on the BCG phagosome but highly enriched on the latex bead phagosome. Immunofluorescence studies confirmed the scarcity of flotillin on BCG phagosomes and demonstrated an inverse correlation between bacterial metabolic activity and flotillin on M. tuberculosis phagosomes. By mass spectrometry, 447 human host proteins were identified on BCG phagosomes, and a partially overlapping set of 289 human proteins on latex bead phagosomes was identified. Interestingly, the majority of the proteins identified consistently on BCG phagosome preparations were also identified on latex bead phagosomes, indicating a high degree of overlap in protein composition of these two compartments. It is likely that many differences in protein composition are quantitative rather than qualitative in nature. Despite the remarkable overlap in protein composition, we consistently identified a number of proteins on the BCG phagosomes that were not identified in any of our latex bead phagosome preparations, including proteins involved in membrane trafficking and signal transduction, such as Ras GTPase-activating-like protein IQGAP1, and proteins of unknown function, such as FAM3C. Our phagosome purification procedure and initial proteomics analyses set the stage for a quantitative comparative analysis of mycobacterial and latex bead phagosome proteomes. PMID- 19815538 TI - Quality assurance in the prospective multi-institutional trial on definitive radiotherapy using high-dose-rate intracavitary brachytherapy for uterine cervical cancer: the individual case review. AB - OBJECTIVE: To assess compliance with the radiotherapy protocol of a multi institutional prospective study (JAROG0401/JROSG04-2), which investigated the efficacy and toxicity of definitive radiotherapy using high-dose-rate intracavitary brachytherapy (HDR-ICBT) for early-stage uterine cervical cancer patients. METHODS: Individual case reviews (ICRs) were performed on all 60 study participants. Radiotherapy data were submitted to the quality assurance (QA) committee, which performed ICRs on 16 QA items according to previously selected criteria. The items focused on quality of external beam radiotherapy (EBRT), HDR ICBT and both. Each item was determined to be either acceptable or a deviation. The QA committee performed ICR three times as planned, two during the patient accrual and the final one just after the final patient accrued. The QA results of the first and second reviews were reported back to the investigators after each ICR. RESULTS: In 40 cases (67%), all 16 QA items were classified as acceptable. One deviation was found in 16 cases, two deviations were identified in 3 cases and three deviations were noted in 1 case. The most frequently observed deviation was missing the rules for determining point A (10 cases). The items described by quantitative values, such as prescribed doses, certain time intervals and overall treatment time, were well followed. The proportion of deviations gradually decreased during the ICR process. CONCLUSIONS: The present ICR demonstrated the favorable radiotherapy compliance with the JAROG0401/JROSG04-2 protocol. The QA process using ICRs can potentially be used to improve the quality of radiotherapy, including HDR-ICBT in the multi-institutional prospective studies for cervical cancer. PMID- 19815539 TI - Right ventricle in pulmonary arterial hypertension: haemodynamics, structural changes, imaging, and proposal of a study protocol aimed to assess remodelling and treatment effects. AB - Although right ventricular (RV) failure is the main cause of death in patients with pulmonary arterial hypertension (PAH), there is insufficient data about the effects of PAH treatment on RV geometry and function mainly because the RV assessment has been hampered by its complex crescentic shape, large infundibulum, and its trabecular nature. Echocardiography is a widely available imaging technique particularly suitable for follow-up studies, because of its non invasive nature, low cost, and lack of ionizing radiation or radioactive agent. Real-time three-dimensional echocardiography (RT3DE) has been shown to be accurate in assessing RV and left ventricular (LV) volumes, stroke volumes, and ejection fractions in comparison with cardiac magnetic resonance imaging. In this review, we describe RV structural and functional changes which occur in patients with PAH and strengths and weaknesses of current non-invasive imaging techniques to assess them. Finally, we describe an ongoing multicentre, prospective observational study involving seven centres expert in treating patients with PAH from four different countries. Investigators will use conventional and advanced echo parameters from RT3DE and speckle-tracking echocardiography to assess the extent of LV and RV remodelling before symptom onset and during pharmacological treatment in patients with PAH. Seventy patients who will survive for at least 1 year will be recruited. All the participating institutions will perform comprehensive standard 2D and Doppler as well as RT3DE examinations with a pre defined imaging protocol. Measurements will be performed at the core echocardiography laboratory by experienced observers who will be unaware of each patient's treatment assignment and whether the examination was a baseline or a follow-up study. Enrolment duration is expected to be 1 year. PMID- 19815540 TI - Adrenergic stimulation increases repolarization dispersion and reduces activation repolarization coupling along the RV endocardium of patients with cardiomyopathy. AB - AIMS: Dispersion of repolarization (DOR) in the human heart is minimized by activation-repolarization coupling. Adrenergic stimulation can be proarrhythmic in patients with impaired left-ventricular function and its effect on repolarization dispersion has not been systematically investigated. Our objective was to study the effect of dobutamine on repolarization dispersion and activation repolarization coupling in patients with cardiomyopathy. METHODS AND RESULTS: Activation recovery intervals (ARI) and activation times (AT) were measured from unipolar electrograms at 10 sites along the apicobasal right ventricle (RV) in 14 patients with cardiomyopathy (LVEF < 40%). These measurements were made during control, dobutamine 2.5-5.0 microg/kg/min, and a recontrol phase while maintaining constant heart rates with atrial pacing. Dispersion of repolarization was calculated from the total recovery time (TRT, AT+ARI). Activation repolarization coupling was assessed by linear regression of ARI and AT. Dispersion of repolarization across all 10 sites and between adjacent sites increased with dobutamine compared with control (whole DOR: range 15 +/- 2 vs. 12 +/- 2 ms, P = 0.06 and standard deviation 5.5 +/- 0.9 vs. 4.3 +/- 0.9 ms, P = 0.04; adjacent DOR: 5.9 +/- 0.8 vs. 4.5 +/- 0.6 ms, P = 0.04). This was associated with shallower ARI/AT slopes (-0.3 +/- 0.2 vs. -0.8 +/- 0.2, P = 0.05) and a decrease in ARI-AT correlation (R(2) 0.4 +/- 0.1 vs. 0.6 +/- 0.1, P = 0.05) with dobutamine compared with control. CONCLUSION: Adrenergic stimulation increases apicobasal RV DOR and reduces coupling between activation and repolarization in patients with cardiomyopathy. This may provide a mechanism for the proarrhythmic potential of heightened adrenergic states in these patients. PMID- 19815541 TI - TRAF2 must bind to cellular inhibitors of apoptosis for tumor necrosis factor (tnf) to efficiently activate nf-{kappa}b and to prevent tnf-induced apoptosis. AB - Tumor necrosis factor (TNF) receptor-associated factor-2 (TRAF2) binds to cIAP1 and cIAP2 (cIAP1/2) and recruits them to the cytoplasmic domain of several members of the TNF receptor (TNFR) superfamily, including the TNF-TNFR1 ligand receptor complex. Here, we define a cIAP1/2-interacting motif (CIM) within the TRAF-N domain of TRAF2, and we use TRAF2 CIM mutants to determine the role of TRAF2 and cIAP1/2 individually, and the TRAF2-cIAP1/2 interaction, in TNFR1 dependent signaling. We show that both the TRAF2 RING domain and the TRAF2 CIM are required to regulate NF-kappaB-inducing kinase stability and suppress constitutive noncanonical NF-kappaB activation. Conversely, following TNFR1 stimulation, cells bearing a CIM-mutated TRAF2 showed reduced canonical NF-kappaB activation and TNF-induced RIPK1 ubiquitylation. Remarkably, the RING domain of TRAF2 was dispensable for these functions. However, like the TRAF2 CIM, the RING domain of TRAF2 was required for protection against TNF-induced apoptosis. These results show that TRAF2 has anti-apoptotic signaling roles in addition to promoting NF-kappaB signaling and that efficient activation of NF-kappaB by TNFR1 requires the recruitment of cIAP1/2 by TRAF2. PMID- 19815542 TI - Structure-based mechanism of CMP-2-keto-3-deoxymanno-octulonic acid synthetase: convergent evolution of a sugar-activating enzyme with DNA/RNA polymerases. AB - The enzyme CMP-Kdo synthetase (KdsB) catalyzes the addition of 2-keto-3 deoxymanno-octulonic acid (Kdo) to CTP to form CMP-Kdo, a key reaction in the biosynthesis of lipopolysaccharide. The reaction catalyzed by KdsB and the related CMP-acylneuraminate synthase is unique among the sugar-activating enzymes in that the respective sugars are directly coupled to a cytosine monophosphate. Using inhibition studies, in combination with isothermal calorimetry, we show the substrate analogue 2beta-deoxy-Kdo to be a potent competitive inhibitor. The ligand-free Escherichia coli KdsB and ternary complex KdsB-CTP-2beta-deoxy-Kdo crystal structures reveal that Kdo binding leads to active site closure and repositioning of the CTP phosphates and associated Mg(2+) ion (Mg-B). Both ligands occupy conformations compatible with an S(n)2-type attack on the alpha phosphate by the Kdo 2-hydroxyl group. Based on strong similarity with DNA/RNA polymerases, both in terms of overall chemistry catalyzed as well as active site configuration, we postulate a second Mg(2+) ion (Mg-A) is bound by the catalytically competent KdsB-CTP-Kdo ternary complex. Modeling of this complex reveals the Mg-A coordinated to the conserved Asp(100) and Asp(235) in addition to the CTP alpha-phosphate and both the Kdo carboxylic and 2-hydroxyl groups. EPR measurements on the Mn(2+)-substituted ternary complex support this model. We propose the KdsB/CNS sugar-activating enzymes catalyze the formation of activated sugars, such as the abundant CMP-5-N-acetylneuraminic acid, by recruitment of two Mg(2+) to the active site. Although each metal ion assists in correct positioning of the substrates and activation of the alpha-phosphate, Mg-A is responsible for activation of the sugar-hydroxyl group. PMID- 19815543 TI - Phosphorylation of protease-activated receptor-2 differentially regulates desensitization and internalization. AB - Protease-activated receptor 2 (PAR2) is a G protein-coupled receptor irreversibly activated by extracellular proteases. Activated PAR2 couples to multiple heterotrimeric G-protein subtypes including G alpha(q), G alpha(i), and G alpha(12/13). Most activated G protein-coupled receptors are rapidly desensitized and internalized following phosphorylation and beta-arrestin binding. However, the role of phosphorylation in regulation of PAR2 signaling and trafficking is not known. To investigate the function of phosphorylation, we generated a PAR2 mutant in which all serines and threonines in the C-tail were converted to alanines and designated it PAR2 0P. In mammalian cells, the addition of agonist induced a rapid and robust increase in phosphorylation of wild-type PAR2 but not the 0P mutant, suggesting that the major sites of phosphorylation occur within the C-tail domain. Moreover, desensitization of PAR2 0P signaling was markedly impaired compared with the wild-type receptor. Wild-type phosphorylated PAR2 internalized through a canonical dynamin, clathrin- and beta-arrestin-dependent pathway. Strikingly, PAR2 0P mutant internalization proceeded through a dynamin dependent but clathrin- and beta-arrestin-independent pathway in both a constitutive and agonist-dependent manner. Collectively, our studies show that PAR2 phosphorylation is essential for beta-arrestin binding and uncoupling from heterotrimeric G-protein signaling and that the presence of serine and threonine residues in the PAR2 C-tail hinder constitutive internalization through a non canonical pathway. Thus, our studies reveal a novel function for phosphorylation that differentially regulates PAR2 desensitization and endocytic trafficking. PMID- 19815544 TI - Regulated endoplasmic reticulum-associated degradation of a polytopic protein: p97 recruits proteasomes to Insig-1 before extraction from membranes. AB - Polytopic membrane proteins subjected to endoplasmic reticulum (ER)-associated degradation are extracted from membranes and targeted to proteasomes for destruction. The extraction mechanism is poorly understood. One polytopic ER protein subjected to ER-associated degradation is Insig-1, a negative regulator of cholesterol synthesis. Insig-1 is rapidly degraded by proteasomes when cells are depleted of cholesterol, and its degradation is inhibited when sterols accumulate in cells. Insig-2, a functional homologue of Insig-1, is degraded slowly, and its degradation is not regulated by sterols. Here, we report that a single amino acid substitution in Insig-2, Insig-2(L210A), causes Insig-2 to be degraded in an accelerated and sterol-regulated manner similar to Insig-1. In seeking an explanation for the accelerated degradation, we found that proteasomes bind to wild type Insig-1 and mutant Insig-2(L210A) but not to wild type Insig-2, whereas the proteins are still embedded in cell membranes. This binding depends on at least two factors, ubiquitination of Insig and association with the ATPase p97/VCP complex. These data suggest that p97 recruits proteasomes to polytopic ER proteins even before they are extracted from membranes. PMID- 19815545 TI - G protein-coupled receptor kinase-2 constitutively regulates D2 dopamine receptor expression and signaling independently of receptor phosphorylation. AB - We investigated the regulatory effects of GRK2 on D(2) dopamine receptor signaling and found that this kinase inhibits both receptor expression and functional signaling in a phosphorylation-independent manner, apparently through different mechanisms. Overexpression of GRK2 was found to suppress receptor expression at the cell surface and enhance agonist-induced internalization, whereas short interfering RNA knockdown of endogenous GRK2 led to an increase in cell surface receptor expression and decreased agonist-mediated endocytosis. These effects were not due to GRK2-mediated phosphorylation of the D(2) receptor as a phosphorylation-null receptor mutant was regulated similarly, and overexpression of a catalytically inactive mutant of GRK2 produced the same effects. The suppression of receptor expression is correlated with constitutive association of GRK2 with the receptor complex as we found that GRK2 and several of its mutants were able to co-immunoprecipitate with the D(2) receptor. Agonist pretreatment did not enhance the ability of GRK2 to co-immunoprecipitate with the receptor. We also found that overexpression of GRK2 attenuated the functional coupling of the D(2) receptor and that this activity required the kinase activity of GRK2 but did not involve receptor phosphorylation, thus suggesting the involvement of an additional GRK2 substrate. Interestingly, we found that the suppression of functional signaling also required the G betagamma binding activity of GRK2 but did not involve the GRK2 N-terminal RH domain. Our results suggest a novel mechanism by which GRK2 negatively regulates G protein-coupled receptor signaling in a manner that is independent of receptor phosphorylation. PMID- 19815546 TI - Lysophosphatidylcholine activates adipocyte glucose uptake and lowers blood glucose levels in murine models of diabetes. AB - Glucose homeostasis is maintained by the orchestration of peripheral glucose utilization and hepatic glucose production, mainly by insulin. In this study, we found by utilizing a combined parallel chromatography mass profiling approach that lysophosphatidylcholine (LPC) regulates glucose levels. LPC was found to stimulate glucose uptake in 3T3-L1 adipocytes dose- and time-dependently, and this activity was found to be sensitive to variations in acyl chain lengths and to polar head group types in LPC. Treatment with LPC resulted in a significant increase in the level of GLUT4 at the plasma membranes of 3T3-L1 adipocytes. Moreover, LPC did not affect IRS-1 and AKT2 phosphorylations, and LPC-induced glucose uptake was not influenced by pretreatment with the PI 3-kinase inhibitor LY294002. However, glucose uptake stimulation by LPC was abrogated both by rottlerin (a protein kinase Cdelta inhibitor) and by the adenoviral expression of dominant negative protein kinase Cdelta. In line with its determined cellular functions, LPC was found to lower blood glucose levels in normal mice. Furthermore, LPC improved blood glucose levels in mouse models of type 1 and 2 diabetes. These results suggest that an understanding of the mode of action of LPC may provide a new perspective of glucose homeostasis. PMID- 19815547 TI - HxcQ liposecretin is self-piloted to the outer membrane by its N-terminal lipid anchor. AB - Secretins are an unusual and important class of bacterial outer membrane (OM) proteins. They are involved in the transport of single proteins or macromolecular structures such as pili, needle complexes, and bacteriophages across the OM. Secretins are multimeric ring-shaped structures that form large pores in the OM. The targeting of such macromolecular structures to the OM often requires special assistance, conferred by specific pilotins or pilot proteins. Here, we investigated HxcQ, the OM component of the second Pseudomonas aeruginosa type II secretion system. We found that HxcQ forms high molecular mass structures resistant to heat and SDS, revealing its secretin nature. Interestingly, we showed that HxcQ is a lipoprotein. Construction of a recombinant nonlipidated HxcQ (HxcQnl) revealed that lipidation is essential for HxcQ function. Further phenotypic analysis indicated that HxcQnl accumulates as multimers in the inner membrane of P. aeruginosa, a typical phenotype observed for secretins in the absence of their cognate pilotin. Our observations led us to the conclusion that the lipid anchor of HxcQ plays a pilotin role. The self-piloting of HxcQ to the OM was further confirmed by its correct multimeric OM localization when expressed in the heterologous host Escherichia coli. Altogether, our results reveal an original and unprecedented pathway for secretin transport to the OM. PMID- 19815548 TI - Calnexin phosphorylation attenuates the release of partially misfolded alpha1 antitrypsin to the secretory pathway. AB - Calnexin is a type I integral membrane phosphoprotein resident of the endoplasmic reticulum. Its intraluminal domain has been deduced to function as a lectin chaperone coordinating the timing of folding of newly synthesized N-linked glycoproteins of the secretory pathway. Its C-terminal cytosolic oriented extension has an ERK1 phosphorylation site at Ser(563) affecting calnexin association with the translocon. Here we find an additional function for calnexin phosphorylation at Ser(563) in endoplasmic reticulum quality control. A low dose of the misfolding agent l-azetidine 2-carboxylic acid slows glycoprotein maturation and diminishes the extent and rate of secretion of newly synthesized secretory alpha1-antitrypsin. Under these conditions the phosphorylation of calnexin is enhanced at Ser(563). Inhibition of this phosphorylation by the MEK1 inhibitor PD98059 enhanced the extent and rate of alpha1-antitrypsin secretion comparable with that achieved by inhibiting alpha-mannosidase activity with kifunensine. This is the first report in which the phosphorylation of calnexin is linked to the efficiency of secretion of a cargo glycoprotein. PMID- 19815549 TI - Meckel-Gruber syndrome protein MKS3 is required for endoplasmic reticulum associated degradation of surfactant protein C. AB - Autosomal dominant mutations in the SFTPC gene are associated with idiopathic pulmonary fibrosis, a progressive lethal interstitial lung disease. Mutations that cause misfolding of the encoded proprotein surfactant protein C (SP-C) trigger endoplasmic reticulum (ER)-associated degradation, a pathway that segregates terminally misfolded substrate for retrotranslocation to the cytosol and degradation by proteasome. Microarray screens for genes involved in SP-C ER associated degradation identified MKS3/TMEM67, a locus previously linked to the ciliopathy Meckel-Gruber syndrome. In this study, MKS3 was identified as a membrane glycoprotein predominantly localized to the ER. Expression of MKS3 was up-regulated by genetic or pharmacological inducers of ER stress. The ER lumenal domain of MKS3 interacted with a complex that included mutant SP-C and associated chaperones, whereas the region predicted to encode the transmembrane domains of MKS3 interacted with cytosolic p97. Deletion of the transmembrane and cytosolic domains abrogated interaction of MKS3 with p97 and resulted in accumulation of mutant SP-C proprotein; knockdown of MKS3 also inhibited degradation of mutant SP C. These results support a model in which MKS3 links the ER lumenal quality control machinery with the cytosolic degradation apparatus. PMID- 19815550 TI - Anionic lipids allosterically modulate multiple nicotinic acetylcholine receptor conformational equilibria. AB - Anionic lipids influence the ability of the nicotinic acetylcholine receptor to gate open in response to neurotransmitter binding, but the underlying mechanisms are poorly understood. We show here that anionic lipids with relatively small headgroups, and thus the greatest ability to influence lipid packing/bilayer physical properties, are the most effective at stabilizing an agonist-activatable receptor. The differing abilities of anionic lipids to stabilize an activatable receptor stem from differing abilities to preferentially favor resting over both uncoupled and desensitized conformations. Anionic lipids thus modulate multiple acetylcholine receptor conformational equilibria. Our data suggest that both lipids and membrane physical properties act as classic allosteric modulators influencing function by interacting with and thus preferentially stabilizing different native acetylcholine receptor conformational states. PMID- 19815551 TI - Non-redundant roles of phosphoinositide 3-kinase isoforms alpha and beta in glycoprotein VI-induced platelet signaling and thrombus formation. AB - Platelets are activated by adhesion to vascular collagen via the immunoglobulin receptor, glycoprotein VI (GPVI). This causes potent signaling toward activation of phospholipase Cgamma2, which bears similarity to the signaling pathway evoked by T- and B-cell receptors. Phosphoinositide 3-kinase (PI3K) plays an important role in collagen-induced platelet activation, because this activity modulates the autocrine effects of secreted ADP. Here, we identified the PI3K isoforms directly downstream of GPVI in human and mouse platelets and determined their role in GPVI dependent thrombus formation. The targeting of platelet PI3Kalpha or -beta strongly and selectively suppressed GPVI-induced Ca(2+) mobilization and inositol 1,4,5-triphosphate production, thus demonstrating enhancement of phospholipase Cgamma2 by PI3Kalpha/beta. That PI3Kalpha and -beta have a non-redundant function in GPVI-induced platelet activation and thrombus formation was concluded from measurements of: (i) serine phosphorylation of Akt, (ii) dense granule secretion, (iii) intracellular Ca(2+) increases and surface expression of phosphatidylserine under flow, and (iv) thrombus formation, under conditions where PI3Kalpha/beta was blocked or p85alpha was deficient. In contrast, GPVI-induced platelet activation was insensitive to inhibition or deficiency of PI3Kdelta or -gamma. Furthermore, PI3Kalpha/beta, but not PI3Kgamma, contributed to GPVI-induced Rap1b activation and, surprisingly, also to Rap1b-independent platelet activation via GPVI. Together, these findings demonstrate that both PI3Kalpha and -beta isoforms are required for full GPVI-dependent platelet Ca(2+) signaling and thrombus formation, partly independently of Rap1b. This provides a new mechanistic explanation for the anti-thrombotic effect of PI3K inhibition and makes PI3Kalpha an interesting new target for anti-platelet therapy. PMID- 19815552 TI - Low density lipoprotein receptor-related protein 1 promotes anti-apoptotic signaling in neurons by activating Akt survival pathway. AB - The low density lipoprotein receptor-related protein 1 (LRP1) is a multi-ligand receptor abundantly expressed in neurons. Previous work has shown that brain LRP1 levels are decreased during aging and in Alzheimer disease. Although mounting evidence has demonstrated a role for LRP1 in the metabolism of apolipoprotein E/lipoprotein and amyloid-beta peptide, whether LRP1 also plays a direct role in neuronal survival is not clear. Here, we show that LRP1 expression is critical for the survival of primary neurons under stress conditions including trophic withdrawal, the presence of apoptosis inducers, or amyloid-beta-induced neurotoxicity. Using lentiviral short hairpin RNA to knock down endogenous LRP1 expression, we showed that a depletion of LRP1 leads to an activation of caspase 3 and increased neuronal apoptosis, an effect that was rescued by a caspase-3 inhibitor. A correlation between decreased Akt phosphorylation and the activation of caspase-3 was demonstrated in LRP1 knocked down neurons. Notably, LRP1 knockdown decreased insulin receptor levels in primary neurons, suggesting that decreased neuronal survival might be a consequence of an impaired insulin receptor signaling pathway. Correspondingly, both insulin receptor and phospho Akt levels were decreased in LRP1 forebrain knock-out mice. These results demonstrate that LRP1 mediates anti-apoptotic function in neurons by regulating insulin receptor and the Akt survival pathway and suggest that restoring LRP1 expression in Alzheimer disease brain might be beneficial to inhibiting neurodegeneration. PMID- 19815553 TI - Suboptimal activation of protease-activated receptors enhances alpha2beta1 integrin-mediated platelet adhesion to collagen. AB - Thrombin and fibrillar collagen are potent activators of platelets at sites of vascular injury. Both agonists cause platelet shape change, granule secretion, and aggregation to form the primary hemostatic plug. Human platelets express two thrombin receptors, protease-activated receptors 1 and 4 (PAR1 and PAR4) and two collagen receptors, the alpha(2)beta(1) integrin (alpha(2)beta(1)) and the glycoprotein VI (GPVI)/FcRgamma chain complex. Although these receptors and their signaling mechanisms have been intensely studied, it is not known whether and how these receptors cooperate in the hemostatic function of platelets. This study examined cooperation between the thrombin and collagen receptors in platelet adhesion by utilizing a collagen-related peptide (alpha2-CRP) containing the alpha(2)beta(1)-specific binding motif, GFOGER, in conjunction with PAR activating peptides. We demonstrate that platelet adhesion to alpha2-CRP is substantially enhanced by suboptimal PAR activation (agonist concentrations that do not stimulate platelet aggregation) using the PAR4 agonist peptide and thrombin. The enhanced adhesion induced by suboptimal PAR4 activation was alpha(2)beta(1)-dependent and GPVI/FcRgamma-independent as revealed in experiments with alpha(2)beta(1)- or FcRgamma-deficient mouse platelets. We further show that suboptimal activation of other platelet G(q)-linked G protein coupled receptors (GPCRs) produces enhanced platelet adhesion to alpha2-CRP. The enhanced alpha(2)beta(1)-mediated platelet adhesion is controlled by phospholipase C (PLC), but is not dependent on granule secretion, activation of alpha(IIb)beta(3) integrin, or on phosphoinositol-3 kinase (PI3K) activity. In conclusion, we demonstrate a platelet priming mechanism initiated by suboptimal activation of PAR4 or other platelet G(q)-linked GPCRs through a PLC-dependent signaling cascade that promotes enhanced alpha(2)beta(1) binding to collagens containing GFOGER sites. PMID- 19815554 TI - Formin-like 1 (FMNL1) is regulated by N-terminal myristoylation and induces polarized membrane blebbing. AB - The formin protein formin-like 1 (FMNL1) is highly restrictedly expressed in hematopoietic lineage-derived cells and has been previously identified as a tumor associated antigen. However, function and regulation of FMNL1 are not well defined. We have identified a novel splice variant (FMNL1gamma) containing an intron retention at the C terminus affecting the diaphanous autoinhibitory domain (DAD). FMNL1gamma is specifically located at the cell membrane and cortex in diverse cell lines. Similar localization of FMNL1 was observed for a mutant lacking the DAD domain (FMNL1DeltaDAD), indicating that deregulation of autoinhibition is effective in FMNL1gamma. Expression of both FMNL1gamma and FMNL1DeltaDAD induces polarized nonapoptotic blebbing that is dependent on N terminal myristoylation of FMNL1 but independent of Src and ROCK activity. Thus, our results describe N-myristoylation as a regulative mechanism of FMNL1 responsible for membrane trafficking potentially involved in a diversity of polarized processes of hematopoietic lineage-derived cells. PMID- 19815555 TI - The deubiquitinating enzyme BAP1 regulates cell growth via interaction with HCF 1. AB - The deubiquitinating enzyme BRCA1-associated protein 1 (BAP1) possesses growth inhibitory activity and functions as a tumor suppressor. In this study we report that BAP1 also plays positive roles in cell proliferation. BAP1 depletion by RNAi inhibits cell proliferation as does overexpression of a dominant negative mutant of BAP1. Mass spectrometry analyses of copurified proteins revealed that BAP1 is associated with factors involved in chromatin modulation and transcriptional regulation. We show that the interaction with host cell factor-1 (HCF-1), a cell cycle regulator composed of HCF-1N and HCF-1C, is critical for the BAP1-mediated growth regulation. We found that HCF-1N is modified with Lys-48-linked polyubiquitin chains on its Kelch domain. The HCF-1 binding motif of BAP1 is required for interaction with HCF-1N and mediates deubiquitination of HCF-1N by BAP1. The importance of the BAP1-HCF-1 interaction is underscored by the fact that growth suppression by the dominant negative BAP1 mutant is entirely dependent on the HCF-1 binding motif. These results suggest that BAP1 regulates cell proliferation by deubiquitinating HCF-1. PMID- 19815556 TI - Down-regulation of seladin-1 increases BACE1 levels and activity through enhanced GGA3 depletion during apoptosis. AB - Seladin-1 is a neuroprotective protein selectively down-regulated in brain regions affected in Alzheimer disease (AD). Seladin-1 protects cells against beta amyloid (Abeta) peptide 42- and oxidative stress-induced apoptosis activated by caspase-3, a key mediator of apoptosis. Here, we have employed RNA interference to assess the molecular effects of seladin-1 down-regulation on the beta secretase (BACE1) function and beta-amyloid precursor protein (APP) processing in SH-SY5Y human neuroblastoma cells in both normal and apoptotic conditions. Our results show that approximately 60% reduction in seladin-1 protein levels, resembling the decrease observed in AD brain, did not significantly affect APP processing or Abeta secretion in normal growth conditions. However, under apoptosis, seladin-1 small interfering RNA (siRNA)-transfected cells showed increased caspase-3 activity on average by 2-fold when compared with control siRNA-transfected cells. Increased caspase-3 activity coincided with a significant depletion of the BACE1-sorting protein, GGA3 (Golgi-localized gamma ear-containing ADP-ribosylation factor-binding protein), and subsequently augmented BACE1 protein levels and activity. Augmented BACE1 activity in turn correlated with the enhanced beta-amyloidogenic processing of APP and ultimately increased Abeta production. These adverse changes associated with decreased cell viability in seladin-1 siRNA-transfected cells under apoptosis. No changes in GGA3 or BACE1 levels were found after seladin-1 knockdown in normal growth conditions. Collectively, our results suggest that under stress conditions, reduced seladin-1 expression results in enhanced GGA3 depletion, which further leads to augmented post-translational stabilization of BACE1 and increased beta amyloidogenic processing of APP. These mechanistic findings related to seladin-1 down-regulation are important in the context of AD as the oxidative stress induced apoptosis plays a key role in the disease pathogenesis. PMID- 19815557 TI - Cytoplasmic ACK1 interaction with multiple receptor tyrosine kinases is mediated by Grb2: an analysis of ACK1 effects on Axl signaling. AB - ACK1 (activated Cdc42-associated kinase 1), a cytoplsmic tyrosine kinase, is implicated in metastatic behavior, cell spreading and migration, and epidermal growth factor receptor (EGFR) signaling. The function of ACK1 in the regulation of receptor tyrosine kinases requires a C-terminal region that demonstrates a significant homology to the EGFR binding domain of MIG6. In this study, we have identified additional receptor tyrosine kinases, including Axl, leukocyte tyrosine kinase, and anaplastic lymphoma kinase, that can bind to the ACK1/MIG6 homology region. Unlike the interaction between MIG6 and EGFR, our data suggest that these receptor tyrosine kinases require the adaptor protein Grb2 for efficient binding, which interacts with highly conserved proline-rich regions that are conserved between ACK1 and MIG6. We have focused on Axl and compared how ACK1/Axl differs from the ACK1/EGFR axis by investigating effects of knockdown of endogenous ACK1. Although EGFR activation promotes ACK1 turnover, Axl activation by GAS6 does not; interestingly, the reciprocal down-regulation of GAS6 stimulated Axl is blocked by removing ACK1. Thus, ACK1 functions in part to control Axl receptor levels. Silencing of ACK1 also leads to diminished ruffling and migration in DU145 and COS7 cells upon GAS6-Axl signaling. The ability of ACK1 to modulate Axl and perhaps anaplastic lymphoma kinase (altered in anaplastic large cell lymphomas) might explain why ACK1 can promote metastatic and transformed behavior in a number of cancers. PMID- 19815558 TI - Features of subunit NuoM (ND4) in Escherichia coli NDH-1: TOPOLOGY AND IMPLICATION OF CONSERVED GLU144 FOR COUPLING SITE 1. AB - The bacterial H(+)-pumping NADH-quinone oxidoreductase (NDH-1) is an L-shaped membrane-bound enzymatic complex. Escherichia coli NDH-1 is composed of 13 subunits (NuoA-N). NuoM (ND4) subunit is one of the hydrophobic subunits that constitute the membrane arm of NDH-1 and was predicted to bear 14 helices. We attempted to clarify the membrane topology of NuoM by the introduction of histidine tags into different positions by chromosomal site-directed mutagenesis. From the data, we propose a topology model containing 12 helices (helices I-IX and XII-XIV) located in transmembrane position and two (helices X and XI) present in the cytoplasm. We reported previously that residue Glu(144) of NuoM was located in the membrane (helix V) and was essential for the energy-coupling activities of NDH-1 (Torres-Bacete, J., Nakamaru-Ogiso, E., Matsuno-Yagi, A., and Yagi, T. (2007) J. Biol. Chem. 282, 36914-36922). Using mutant E144A, we studied the effect of shifting the glutamate residue to all sites within helix V and three sites each in helix IV and VI on the function of NDH-1. Twenty double site directed mutants including the mutation E144A were constructed and characterized. None of the mutants showed alteration in the detectable levels of expressed NuoM or on the NDH-1 assembly. In addition, most of the double mutants did not restore the energy transducing NDH-1 activities. Only two mutants E144A/F140E and E144A/L147E, one helix turn downstream and upstream restored the energy transducing activities of NDH-1. Based on these results, a role of Glu(144) for proton translocation has been discussed. PMID- 19815559 TI - Molecular basis of glucagon-like peptide 1 docking to its intact receptor studied with carboxyl-terminal photolabile probes. AB - The glucagon-like peptide 1 (GLP1) receptor is a member of Family B G protein coupled receptors and represents an important drug target for type 2 diabetes. Despite recent solution of the structure of the amino-terminal domain of this receptor and that of several close family members, understanding of the molecular basis of natural ligand GLP1 binding to its intact receptor remains limited. The goal of this study was to explore spatial approximations between specific receptor residues within the carboxyl terminus of GLP1 and its receptor as normally docked. Therefore, we developed and characterized two high affinity, full-agonist photolabile GLP1 probes having sites for covalent attachment in positions 24 and 35. Both probes labeled the receptor specifically and saturably. Subsequent peptide mapping using chemical and proteinase cleavages of purified wild-type and mutant GLP1 receptor identified that the Arg(131)-Lys(136) segment at the juxtamembrane region of the receptor amino terminus contained the site of labeling for the position 24 probe, and the specific receptor residue labeled by this probe was identified as Glu(133) by radiochemical sequencing. Similarly, nearby residue Glu(125) within the same region of the receptor amino-terminal domain was identified as the site of labeling by the position 35 probe. These data represent the first direct demonstration of spatial approximation between GLP1 and its intact receptor as docked, providing two important constraints for the modeling of this interaction. This should expand our understanding of the molecular basis of natural agonist ligand binding to the GLP1 receptor and may be relevant to other family members. PMID- 19815560 TI - TRPC5 is a Ca2+-activated channel functionally coupled to Ca2+-selective ion channels. AB - TRPC5 forms non-selective cation channels. Here we studied the role of internal Ca(2+) in the activation of murine TRPC5 heterologously expressed in human embryonic kidney cells. Cell dialysis with various Ca(2+) concentrations (Ca(2+)(i)) revealed a dose-dependent activation of TRPC5 channels by internal Ca(2+) with EC(50) of 635.1 and 358.2 nm at negative and positive membrane potentials, respectively. Stepwise increases of Ca(2+)(i) induced by photolysis of caged Ca(2+) showed that the Ca(2+) activation of TRPC5 channels follows a rapid exponential time course with a time constant of 8.6 +/- 0.2 ms at Ca(2+)(i) below 10 microM, suggesting that the action of internal Ca(2+) is a primary mechanism in the activation of TRPC5 channels. A second slow activation phase with a time to peak of 1.4 +/- 0.1 s was also observed at Ca(2+)(i) above 10 microM. In support of a Ca(2+)-activation mechanism, the thapsigargin-induced release of Ca(2+) from internal stores activated TRPC5 channels transiently, and the subsequent Ca(2+) entry produced a sustained TRPC5 activation, which in turn supported a long-lasting membrane depolarization. By co-expressing STIM1 plus ORAI1 or the alpha(1)C and beta(2) subunits of L-type Ca(2+) channels, we found that Ca(2+) entry through either calcium-release-activated-calcium or voltage dependent Ca(2+) channels is sufficient for TRPC5 channel activation. The Ca(2+) entry activated TRPC5 channels under buffering of internal Ca(2+) with EGTA but not with BAPTA. Our data support the hypothesis that TRPC5 forms Ca(2+)-activated cation channels that are functionally coupled to Ca(2+)-selective ion channels through local Ca(2+) increases beneath the plasma membrane. PMID- 19815561 TI - Crystal structures of progressive Ca2+ binding states of the Ca2+ sensor Ca2+ binding domain 1 (CBD1) from the CALX Na+/Ca2+ exchanger reveal incremental conformational transitions. AB - Na(+)/Ca(2+) exchangers (NCX) constitute a major Ca(2+) export system that facilitates the re-establishment of cytosolic Ca(2+) levels in many tissues. Ca(2+) interactions at its Ca(2+) binding domains (CBD1 and CBD2) are essential for the allosteric regulation of Na(+)/Ca(2+) exchange activity. The structure of the Ca(2+)-bound form of CBD1, the primary Ca(2+) sensor from canine NCX1, but not the Ca(2+)-free form, has been reported, although the molecular mechanism of Ca(2+) regulation remains unclear. Here, we report crystal structures for three distinct Ca(2+) binding states of CBD1 from CALX, a Na(+)/Ca(2+) exchanger found in Drosophila sensory neurons. The fully Ca(2+)-bound CALX-CBD1 structure shows that four Ca(2+) atoms bind at identical Ca(2+) binding sites as those found in NCX1 and that the partial Ca(2+) occupancy and apoform structures exhibit progressive conformational transitions, indicating incremental regulation of CALX exchange by successive Ca(2+) binding at CBD1. The structures also predict that the primary Ca(2+) pair plays the main role in triggering functional conformational changes. Confirming this prediction, mutagenesis of Glu(455), which coordinates the primary Ca(2+) pair, produces dramatic reductions of the regulatory Ca(2+) affinity for exchange current, whereas mutagenesis of Glu(520), which coordinates the secondary Ca(2+) pair, has much smaller effects. Furthermore, our structures indicate that Ca(2+) binding only enhances the stability of the Ca(2+) binding site of CBD1 near the hinge region while the overall structure of CBD1 remains largely unaffected, implying that the Ca(2+) regulatory function of CBD1, and possibly that for the entire NCX family, is mediated through domain interactions between CBD1 and the adjacent CBD2 at this hinge. PMID- 19815562 TI - 'Leaky' ryanodine receptors and sudden cardiac death. PMID- 19815563 TI - Long-term improvement in mdx cardiomyopathy after therapy with peptide-conjugated morpholino oligomers. AB - AIMS: The cardiomyopathy found in Duchenne muscular dystrophy (DMD) is responsible for death due to heart failure in approximately 30% of patients and additionally contributes to many DMD morbidities. Strategies to bypass DMD causing mutations to allow an increase in body-wide dystrophin have proved promising, but increasing cardiac dystrophin continues to be challenging. The purpose of this study was to determine if therapeutic restoration of cardiac dystrophin improved the significant cardiac hypertrophy and diastolic dysfunction identified in X-linked muscular dystrophy (mdx) dystrophin-null mouse due to a truncation mutation over time after treatment. METHODS AND RESULTS: Mice lacking dystrophin due to a truncation mutation (mdx) were given an arginine-rich, cell penetrating, peptide-conjugated phosphorodiamidate morpholino oligomer (PPMO) that delivered a splice-switching oligonucleotide-mediated exon skipping therapy to restore dystrophin in mdx mice before the development of detectable cardiomyopathy. PPMO successfully restored cardiac dystrophin expression, preserved cardiac sarcolemma integrity, and prevented the development of cardiac pathology that develops in mdx-null mice over time. By echocardiography and Doppler analysis of the mitral valve, we identified that PPMO treatment of mdx mice prevented the cardiac hypertrophy and diastolic dysfunction identified in sham-treated, age-matched mdx mice, characteristic of DMD patients early in the disease process, in as little as 5-6 weeks after the initiation of treatment. Surprisingly, despite the short-term replacement of cardiac dystrophin (<1% present after 12 weeks by immunodetection), PPMO therapy also provided a durable cardiac improvement in cardiac hypertrophy and diastolic dysfunction for up to 7 months after the initiation of treatment. CONCLUSION: These results demonstrate for the first time that PPMO-mediated exon skipping therapy early in the course of DMD may effectively prevent or slow down associated cardiac hypertrophy and diastolic dysfunction with significant long-term impact. PMID- 19815564 TI - Activation of SIRT1 by resveratrol induces KLF2 expression conferring an endothelial vasoprotective phenotype. AB - AIMS: Resveratrol activates Sirtuin 1 (SIRT1), a nicotinamide adenine dinucleotide-dependent deacetylase which modulates metabolic homeostasis and improves several pathophysiological features present in diseases of ageing. In particular, it has been shown that SIRT1 activation improves endothelial dysfunction and suppresses vascular inflammation, two central pathophysiological processes involved in the initiation and progression of cardiovascular disease. The downstream targets of SIRT1 activation in this context, however, remain poorly defined. Therefore, in this study, we aimed to characterize mechanistically how SIRT1 activation regulates the endothelial vasoprotective phenotype. METHODS AND RESULTS: We demonstrate that SIRT1 activation by resveratrol increases the expression of the transcription factor Kruppel-like factor 2 (KLF2) in human vascular endothelial cells, resulting in the orchestrated regulation of transcriptional programs critical for conferring an endothelial vasoprotective phenotype. Moreover, we show that KLF2 upregulation by resveratrol occurs via a mitogen-activated protein kinase 5/myocyte enhancing factor 2-dependent signalling pathway. CONCLUSION: Collectively, these observations provide a new mechanistic framework to understand the vascular protective effects mediated by SIRT1 activators and define KLF2 as a critical mediator of these effects. PMID- 19815566 TI - Three strikes--don't die of a broken heart. AB - There are multiple layers of complexity in prevention of vehicle related blunt traumatic aortic rupture (BTAR), many of which are enshrined within government policy and car design. We present a 'layers of protection analysis' (LOPA) based loosely on original work by Professor John Doyle, which describes these attempts to 'design out' the risk of BTAR following a vehicle collision. We have modified this approach to include a physiological dimension suggesting that this may be a factor in susceptibility to aortic injury following trauma. Understanding processes involved in BTAR following vehicle collisions is key to designing preventative processes. PMID- 19815565 TI - On to the road to degradation: atherosclerosis and the proteasome. AB - Protein metabolism is a central element of every living cell. The ubiquitin proteasome system (UPS) is an integral part of the protein metabolism machinery mediating post-transcriptional processing and degradation of the majority of intracellular proteins. Over the past few years, remarkable progress has been made in our understanding of the role of the UPS in vascular biology and pathobiology, particularly atherosclerosis. This review reflects on the recent developments from the effects on endothelial cells and the initial stage of atherosclerosis to the effects on vascular smooth muscle and the progression stage of atherosclerosis and finally to the effects on cell viability and the complication stage of atherosclerosis. It will conclude with the integration of the available information in a synoptic view of the involvement of the UPS in atherosclerosis. PMID- 19815567 TI - Transfusion of red blood cells: the impact on short-term and long-term survival after coronary artery bypass grafting, a ten-year follow-up. AB - Transfusion of red blood cells (RBC) and other blood products in patients undergoing coronary artery bypass grafting (CABG) is associated with increased mortality and morbidity. We retrospectively analyzed data of patients who underwent an isolated coronary bypass graft operation between January 1998 and December 2007. Mean follow-up was 1696+/-1026 days, with exclusion of 122 patients lost to follow-up and 80 patients who received 10 units of RBC. Of the remaining patients, 8001 (76.7%) received no RBC, 1621 (15.2%) received 1-2 units of RBC, 593 (5.7%) received 3-5 units and 220 (2.1%) received 6-10 units. The number of transfused RBC was a predictor for early but not for late mortality. When compared to expected survival, survival of patients not receiving any blood product was better, while survival of patients receiving >3 units of RBC was worse. Transfusion of RBC is an independent, dose-dependent risk factor for early mortality after revascularization. Compared to expected survival, receiving no RBC improves patient long-term survival, whereas receiving three or more units of RBC significantly decreases patient survival. PMID- 19815568 TI - Surgical management of right coronary artery-coronary sinus fistula causing severe mitral and tricuspid regurgitation. AB - Coronary arteriovenous (AV) fistula is a rare congenital anomaly, mostly diagnosed incidentally during routine coronary angiography. We report a symptomatic patient with right coronary artery to coronary sinus (RCA-CS) fistula, complicated by aneurysmal dilatation and thrombosis of the CS, causing severe mitral regurgitation (MR) and tricuspid regurgitation (TR). PMID- 19815569 TI - BcMF9, a novel polygalacturonase gene, is required for both Brassica campestris intine and exine formation. AB - BACKGROUND AND AIMS: The polygalacturonase (PG) gene family has been found to be enriched in pollen of several species; however, little is currently known about the function of the PG gene in pollen development. To investigate the exact role that the PG gene has played in pollen development and about this family in general, one putative PG gene, Brassica campestris Male Fertility 9 (BcMF9), was isolated from Chinese cabbage (Brassica campestris ssp. chinensis, syn. B. rapa ssp. chinensis) and characterized. METHODS: RT-PCR, northern blotting and in situ hybridization were used to analyse the expression pattern of BcMF9, and antisense RNA technology was applied to study the function of this gene. KEY RESULTS: BcMF9 is expressed in particular in the tapetum and microspore during the late stages of pollen development. Antisense RNA transgenic plants that displayed decreased expression of BcMF9 showed pollen morphological defects that resulted in reduced pollen germination efficiency. Transmission electron microscopy revealed that the homogeneous pectic exintine layer of pollen facing the exterior was over developed and predominantly occupied the intine, reversing the normal proportional distribution of the internal endintine layer and the external exintine in transgenic pollen. Inhibition of BcMF9 also resulted in break-up of the previously formed tectum and baculae from the beginning of the binucleate stage, as a result of premature degradation of tapetum. CONCLUSIONS: Several lines of evidence, including patterns of BcMF9 expression and phenotypic defects, suggest a sporophytic role in exine patterning, and a gametophytic mode of action of BcMF9 in intine formation. BcMF9 might act as a co-ordinator in the late stages of tapetum degeneration, and subsequently in the regulation of wall material secretion and, in turn, exine formation. BcMF9 might also play a role in intine formation, possibly via regulation of the dynamic metabolism of pectin. PMID- 19815570 TI - Arbuscular mycorrhizal fungi in alleviation of salt stress: a review. AB - BACKGROUND: Salt stress has become a major threat to plant growth and productivity. Arbuscular mycorrhizal fungi colonize plant root systems and modulate plant growth in various ways. SCOPE: This review addresses the significance of arbuscular mycorrhiza in alleviation of salt stress and their beneficial effects on plant growth and productivity. It also focuses on recent progress in unravelling biochemical, physiological and molecular mechanisms in mycorrhizal plants to alleviate salt stress. CONCLUSIONS: The role of arbuscular mycorrhizal fungi in alleviating salt stress is well documented. This paper reviews the mechanisms arbuscular mycorrhizal fungi employ to enhance the salt tolerance of host plants such as enhanced nutrient acquisition (P, N, Mg and Ca), maintenance of the K(+) : Na(+) ratio, biochemical changes (accumulation of proline, betaines, polyamines, carbohydrates and antioxidants), physiological changes (photosynthetic efficiency, relative permeability, water status, abscissic acid accumulation, nodulation and nitrogen fixation), molecular changes (the expression of genes: PIP, Na(+)/H(+) antiporters, Lsnced, Lslea and LsP5CS) and ultra-structural changes. Theis review identifies certain lesser explored areas such as molecular and ultra-structural changes where further research is needed for better understanding of symbiosis with reference to salt stress for optimum usage of this technology in the field on a large scale. This review paper gives useful benchmark information for the development and prioritization of future research programmes. PMID- 19815571 TI - Genetic analysis of potassium use efficiency in Brassica oleracea. AB - BACKGROUND AND AIMS: Potassium (K) fertilizers are used in intensive and extensive agricultural systems to maximize production. However, there are both financial and environmental costs to K-fertilization. It is therefore important to optimize the efficiency with which K-fertilizers are used. Cultivating crops that acquire and/or utilize K more effectively can reduce the use of K fertilizers. The aim of the present study was to determine the genetic factors affecting K utilization efficiency (KUtE), defined as the reciprocal of shoot K concentration (1/[K](shoot)), and K acquisition efficiency (KUpE), defined as shoot K content, in Brassica oleracea. METHODS: Genetic variation in [K](shoot) was estimated using a structured diversity foundation set (DFS) of 376 accessions and in 74 commercial genotypes grown in glasshouse and field experiments that included phosphorus (P) supply as a treatment factor. Chromosomal quantitative trait loci (QTL) associated with [K](shoot) and KUpE were identified using a genetic mapping population grown in the glasshouse and field. Putative QTL were tested using recurrent backcross substitution lines in the glasshouse. KEY RESULTS: More than two-fold variation in [K](shoot) was observed among DFS accessions grown in the glasshouse, a significant proportion of which could be attributed to genetic factors. Several QTL associated with [K](shoot) were identified, which, despite a significant correlation in [K](shoot) among genotypes grown in the glasshouse and field, differed between these two environments. A QTL associated with [K](shoot) in glasshouse-grown plants (chromosome C7 at 62.2 cM) was confirmed using substitution lines. This QTL corresponds to a segment of arabidopsis chromosome 4 containing genes encoding the K+ transporters AtKUP9, AtAKT2, AtKAT2 and AtTPK3. CONCLUSIONS: There is sufficient genetic variation in B. oleracea to breed for both KUtE and KUpE. However, as QTL associated with these traits differ between glasshouse and field environments, marker-assisted breeding programmes must consider carefully the conditions under which the crop will be grown. PMID- 19815572 TI - Temporal regulation of cell-wall pectin methylesterase and peroxidase isoforms in cadmium-treated flax hypocotyl. AB - BACKGROUND AND AIMS: In hypocotyls of flax (Linum usitatissimum) cadmium-induced reorientation of growth (i.e. an increase in expansion and a decrease in elongation) coincides with marked changes in the methylesterification and cross linking of homogalacturonans within various cell-wall (CW) domains. The aim of the present study was to examine the involvement of pectin methylesterase (PME) and peroxidase (PER) in this cadmium-induced CW remodelling. METHODS: CW proteins were extracted from hypocotyls of 10- and 18-d-old flax that had been treated or not treated with 0.5 mm Cd(NO(3))(2). PME and PER expression within these extracts was detected by LC/MS, by isoelectric focusing and enzyme activity assays. Transcript expression by RT-PCR of known flax PME and PER genes was also measured in corresponding samples. KEY RESULTS: In cadmium-treated seedlings, PME activity increased as compared with controls, particularly at day 10. The increased activity of PME was accompanied by increased abundance of both a basic protein isoform (B2) and a particular transcript (Lupme5). In contrast, induction of PER activity by cadmium was highest at day 18. Among the four reported PER genes, Flxper1 and 3 increased in abundance in the presence of cadmium at day 18. CONCLUSIONS: The temporal regulation of Lupme and Flxper genes and of their respective enzyme activities fits the previously reported cadmium-induced structural changes of homogalacturonans within the CWs. After PME-catalysed de esterification of homogalacturonans, their cross-linking would depend on the activity of PERs interacting with calcium-dimerized blocks and reinforce the cell cohesion during the cadmium-induced swelling. PMID- 19815573 TI - Effects of soy protein and isoflavones on circulating hormone concentrations in pre- and post-menopausal women: a systematic review and meta-analysis. PMID- 19815574 TI - Some aspects of the airborne transmission of infection. AB - The relationship between the human body and the dissemination of potentially pathogenic particles and droplets is described. Airborne transmission of infection in operating theatres and a burns unit and the part played by the human microclimate and its interaction with ventilating air flows is discussed. The mechanisms by which different garment assemblies used for surgery can enhance particle dispersion are illustrated and the way that floor cleaning can increase the concentration of airborne organisms is described. The development of the successful use of ultra-clean air systems in orthopaedic implant surgery is reviewed. Relationships between contact and airborne transmission of disease are explored and ways by which containment strategies and metrics used in pharmaceutical and electronics manufacturing can be applied to the design and monitoring of healthcare areas is discussed. It is suggested that currently available techniques involving architectural, ventilation and operational aspects of healthcare provision, when properly applied, can markedly improve treatment outcomes that may otherwise be compromised by hospital-acquired infections involving both bacteria and viruses. PMID- 19815575 TI - A schlieren optical study of the human cough with and without wearing masks for aerosol infection control. AB - Various infectious agents are known to be transmitted naturally via respiratory aerosols produced by infected patients. Such aerosols may be produced during normal activities by breathing, talking, coughing and sneezing. The schlieren optical method, previously applied mostly in engineering and physics, can be effectively used here to visualize airflows around human subjects in such indoor situations, non-intrusively and without the need for either tracer gas or airborne particles. It accomplishes this by rendering visible the optical phase gradients owing to real-time changes in air temperature. In this study, schlieren video records are obtained of human volunteers coughing with and without wearing standard surgical and N95 masks. The object is to characterize the exhaled airflows and evaluate the effect of these commonly used masks on the fluid dynamic mechanisms that spread infection by coughing. Further, a high-speed schlieren video of a single cough is analysed by a computerized method of tracking individual turbulent eddies, demonstrating the non-intrusive velocimetry of the expelled airflow. Results show that human coughing projects a rapid turbulent jet into the surrounding air, but that wearing a surgical or N95 mask thwarts this natural mechanism of transmitting airborne infection, either by blocking the formation of the jet (N95 mask), or by redirecting it in a less harmful direction (surgical mask). PMID- 19815576 TI - Movement of airborne contaminants in a hospital isolation room. AB - We analyse the characteristics of a force-ventilated isolation room, and the contributions to transport caused by the movement of people and doors opening/closing. The spread of fine droplets and particles can be understood, to leading order, by considering the movement of passive contaminants. A scaled (1:10) model of an isolation room (with water instead of air) was used to analyse the dilution of a passive contaminant (food dye), released either instantaneously or at a constant rate. The high level of turbulence, typical of isolation rooms, ensures that the dye concentration is uniform within the model room and mixing is perfect, and the measured mean concentration can be predicted theoretically. In a second series of experiments, the exchange generated by a door opening/closing is measured for different opening angles. A dipolar vortex is generated at the tip of the door which moves into the centre of the room, with a large coherent structure moving along the wall. The exchange volume is comparable to the swept volume of the door. Larger droplets and particles do not move passively. Their movement within a turbulent flow is studied by combining a Lagrangian model of particle movement with a kinematic simulation of a pseudo turbulent flow. The results show that while the mean fall velocity of particles is largely unchanged, turbulence significantly enhances horizontal and vertical dispersion. The horizontal spread as a function of the level of turbulence and droplet properties is estimated. The conclusions from both studies are brought together and discussed in the context of the airborne spread of contaminants within a general hospital room. PMID- 19815577 TI - Apoptomirs: small molecules have gained the license to kill. AB - Apoptosis is a tightly regulated form of cell death and represents an important process during normal development. In the past years, the scientific community has produced remarkable advances in our understanding of cancer biology, realizing that apoptosis and the genes that control it have a profound effect on the malignant phenotype. Recently, a new class of non-coding RNA genes, known as microRNA (miRNA or miR), have been demonstrated to play important roles in diverse biological processes, including development, cell differentiation, proliferation, and apoptosis. This suggests that other oncogenic mechanisms are needed to produce selective pressure to override apoptosis during multistage carcinogenesis. Intriguingly, since most cytotoxic anticancer agents induce apoptosis, it is possible that defects in apoptotic programs may contribute to treatment failure. Several studies strongly suggest a role for microRNAs in modulating sensitive/resistant phenotypes to cytotoxic therapy, calling for further investigation and validation of microRNA functions and targets in order to improve sensitivity to cancer treatments, thus ultimately improving prognosis and survival. Here, we review the current findings about microRNAs focusing on their involvement in the apoptotic process. PMID- 19815578 TI - Effect of subinhibitory concentrations of benzalkonium chloride on the competitiveness of Pseudomonas aeruginosa grown in continuous culture. AB - This study investigates the link between adaptation to biocides and antibiotics in Pseudomonas aeruginosa. An enrichment continuous culture of P. aeruginosa NCIMB 10421 (MIC 25 mg BKC l(-1)) was operated (D=0.04 h(-1), 792 h) with added benzalkonium chloride (BKC). A derivative, PA-29 (696 h), demonstrated a >12-fold decrease in sensitivity to the biocide (MIC >350 mg BKC l(-1)). The variant demonstrated a 256-fold increase in resistance to ciprofloxacin, with a mutation in the gyrA gene (Thr-83-->Ile). Similarly, culturing of the original strain in a continuous-culture system with ciprofloxacin selection pressure led to the evolution of BKC-adapted populations (MIC 100 mg BKC l(-1)). Efflux pump activity predominantly contributed to the developed phenotype of PA-29. An amino acid substitution (Val-51-->Ala) in nfxB, the Mex efflux system regulator gene, was observed for PA-29. Overexpression of both MexAB-OprM and MexCD-OprJ was recorded for PA-29. Similarly, mexR, a repressor of the Mex system, was downregulated. Competition studies were carried out in continuous culture between PA-29 and the original strain (in the presence of subinhibitory concentrations of BKC). The outcome of competition was influenced by the concentration of biocide used and the nature of limiting nutrient. The inclusion of 1 mg BKC l(-1) in the medium feed was sufficient to select (S=0.011) for the BKC-adapted strain in magnesium limited culture. Conversely, the presence of 10 mg BKC l(-1) in the medium supply was insufficient to select for the same organism (S=-0.017) in the glucose limited culture. These results indicate the importance of environmental conditions on selection and maintenance of biocide adaptation. PMID- 19815579 TI - Defects in flagellin glycosylation affect the virulence of Pseudomonas syringae pv. tabaci 6605. AB - Flagellar motility and its glycosylation are indispensable for the virulence of Pseudomonas syringae pv. tabaci 6605. Six serine residues of the flagellin protein at positions 143, 164, 176, 183, 193 and 201 are glycosylated, and the glycan structure at 201 was determined to consist of a trisaccharide of two L rhamnosyl residues and a modified 4-amino-4,6-dideoxyglucosyl (viosamine) residue. To investigate the glycan structures attached to the other serine residues and to identify the glycans important for virulence, Ser/Ala-substituted mutants were generated. Six mutant strains that each retained a single glycosylated serine residue were generated by replacing five of the six serine residues with alanine residues. MALDI-TOF mass analysis of flagellin proteins revealed that the major component of each glycan was a trisaccharide basically similar to that at position 201, but with heterogeneity in glycoform distribution. Swarming motility and amounts of acylhomoserine lactones (AHLs) as quorum-sensing signal molecules were significantly reduced, especially in the S143-5S/A, S164-5S/A and S201-5S/A mutants, whereas tolerance to antibiotics was increased in these three mutants. All the mutants showed lower ability to cause disease on host tobacco plants. These results supported our previous finding that glycosylation of the most externally located sites on the surface of the flagellin molecule, such as S176 and S183, is required for virulence in P. syringae pv. tabaci 6605. Furthermore, it is speculated that flagellum-dependent motility might be correlated with quorum sensing and antibiotic resistance. PMID- 19815580 TI - The Bam (Omp85) complex is involved in secretion of the autotransporter haemoglobin protease. AB - Autotransporters are large virulence factors secreted by Gram-negative bacteria. They are synthesized with a C-terminal domain that forms a beta-barrel pore in the outer membrane implicated in translocation of the upstream 'passenger' domain across the outer membrane. However, recent structural data suggest that the diameter of the beta-barrel pore is not sufficient to allow the passage of partly folded structures observed for several autotransporters. Here, we have used a stalled translocation intermediate of the autotransporter Hbp to identify components involved in insertion and translocation of the protein across the outer membrane. At this intermediate stage the beta-domain was not inserted and folded as an integral beta-barrel in the outer membrane whereas part of the passenger was surface exposed. The intermediate was copurified with the periplasmic chaperone SurA and subunits of the Bam (Omp85) complex that catalyse the insertion and assembly of outer-membrane proteins. The data suggest a critical role for this general machinery in the translocation of autotransporters across the outer membrane. PMID- 19815581 TI - Incidence of pregnancy after expectant, medical, or surgical management of spontaneous first trimester miscarriage: long term follow-up of miscarriage treatment (MIST) randomised controlled trial. AB - OBJECTIVES: To compare fertility rates after the three methods of managing early miscarriage in women recruited to the MIST (miscarriage treatment) randomised controlled trial. SETTING: Early pregnancy clinics of acute hospitals in the south west region of England. PARTICIPANTS: 1199 women who had had an early miscarriage (<13 weeks) confirmed by scan. INTERVENTION: Expectant, medical, or surgical management. MAIN OUTCOME MEASURES: Self reported pregnancy rates and live birth rates. RESULTS: Of 1199 women recruited to the trial, 1128 consented to follow-up. Of these, 762 women replied giving pregnancy details (68% response rate). Respondents were representative of the trial participants. The live birth rate five years after the index miscarriage was similar in the three management groups: 177/224 (79%, 95% confidence interval 73% to 84%) in the expectant management group, 181/230 (79%, 73% to 84%) in the medical group, and 192/235 (82%, 76% to 86%) in the surgical group. There was also no significant difference according to previous birth history. Older women and those with previous miscarriages were significantly less likely to subsequently give birth. CONCLUSION: Method of miscarriage management does not affect subsequent pregnancy rates with around four in five women giving birth within five years of the index miscarriage. Women can be reassured that long term fertility concerns need not affect their choice of miscarriage management. TRIAL REGISTRATION: National Research Register N0467011677/N0467073587. PMID- 19815582 TI - Cost effectiveness analysis of including boys in a human papillomavirus vaccination programme in the United States. AB - OBJECTIVE: To assess the cost effectiveness of including preadolescent boys in a routine human papillomavirus (HPV) vaccination programme for preadolescent girls. DESIGN: Cost effectiveness analysis from the societal perspective. SETTING: United States. POPULATION: Girls and boys aged 12 years. INTERVENTIONS: HPV vaccination of girls alone and of girls and boys in the context of screening for cervical cancer. Main outcome measure Incremental cost effectiveness ratios, expressed as cost per quality adjusted life year (QALY) gained. RESULTS: With 75% vaccination coverage and an assumption of complete, lifelong vaccine efficacy, routine HPV vaccination of 12 year old girls was consistently less than $50,000 per QALY gained compared with screening alone. Including preadolescent boys in a routine vaccination programme for preadolescent girls resulted in higher costs and benefits and generally had cost effectiveness ratios that exceeded $100,000 per QALY across a range of HPV related outcomes, scenarios for cervical cancer screening, and assumptions of vaccine efficacy and duration. Vaccinating both girls and boys fell below a willingness to pay threshold of $100,000 per QALY only under scenarios of high, lifelong vaccine efficacy against all HPV related diseases (including other non-cervical cancers and genital warts), or scenarios of lower efficacy with lower coverage or lower vaccine costs. CONCLUSIONS: Given currently available information, including boys in an HPV vaccination programme generally exceeds conventional thresholds of good value for money, even under favourable conditions of vaccine protection and health benefits. Uncertainty still exists in many areas that can either strengthen or attenuate our findings. As new information emerges, assumptions and analyses will need to be iteratively revised to continue to inform policies for HPV vaccination. PMID- 19815584 TI - Should we screen low risk patients for meticillin resistant Staphylococcus aureus? PMID- 19815583 TI - Effect of "collaborative requesting" on consent rate for organ donation: randomised controlled trial (ACRE trial). AB - OBJECTIVE: To determine whether collaborative requesting increases consent for organ donation from the relatives of patients declared dead by criteria for brain stem death. DESIGN: Unblinded multicentre randomised controlled trial using a sequential design. Centralised 24 hour telephone randomisation based on randomised permuted blocks of 10. SETTING: 79 general, neuroscience, and paediatric intensive care units in the United Kingdom. PARTICIPANTS: 201 relatives of patients meeting criteria for brain stem death. Relatives were blind to the intervention and to the trial; all other participants were necessarily unblinded. INTERVENTIONS: Collaborative requesting for consent for organ donation by the potential donor's clinician and a donor transplant coordinator (organ procurement officer) compared with routine requesting by the clinical team alone. MAIN OUTCOME MEASURE: Proportion of relatives consenting to organ donation. RESULTS: 101 relatives were randomised to routine requesting and 100 to collaborative requesting. All were analysed on an intention to treat basis. In the routine requesting group, 62 relatives consented to organ donation. In the collaborative requesting group, 57 relatives consented. After correction for the ethnicity, age, and sex of the potential donors the risk adjusted ratio of the odds of consent in the collaborative requesting group relative to the routine group was 0.80 (95% confidence interval 0.43 to 1.53), with a P value of 0.49 adjusted for interim analysis and trial over-running. The conversion rate (donors with consent from whom any organs were retrieved) was 92% (57/62) in the routine requesting group and 79% (45/57) in the collaborative requesting group (P=0.043). There were 140 approaches to relatives in the per protocol analysis, leading to 60.3% (44/73) consent after routine and 67.2% (45/67) after collaborative requesting (risk adjusted odds ratio of consent 1.47, 0.67 to 3.20, P=0.33). CONCLUSION: There is no increase in consent rates for organ donation when collaborative requesting is used in place of routine requesting by the patient's clinician. TRIAL REGISTRATION: ISRCTN01169903. PMID- 19815585 TI - Should HPV vaccine be given to men? PMID- 19815586 TI - Early-onset GH deficiency results in spatial memory impairment in mid-life and is prevented by GH supplementation. AB - GH levels increase to high concentrations immediately before puberty then progressively decline with age. GH deficiency (GHD) originating in childhood is treated with GH supplementation to foster somatic development during adolescence. It is not clear if or how early GH replacement affects memory in adulthood, or whether it can prevent the cognitive deficits commonly observed in adults with childhood-onset GHD. Rats homozygous for the Dw-4 mutation (dwarf) do not exhibit the normal increase in GH at 4 weeks of age when GH levels normally rise and are used to model childhood or early-onset GHD (EOGHD). One group of these rats was injected with GH from 4 to 14 weeks of age to model GH supplementation during adolescence with GHD beginning in adulthood (adult-onset GHD; AOGHD). Another group received GH from 4 weeks throughout the lifespan to model normal lifespan GH (GH-replete). Age-matched, Dw-4 heterozygous rats (HZ) do not express the dwarf phenotype and were used as controls. At 8 and 18 months of age, spatial learning in the water maze was assessed. At 8 months of age all experimental groups were equally proficient. However, at 18 months of age, the EOGHD group had poor spatial learning compared to the AOGHD, GH-replete, and HZ groups. Our data indicate that GHD during adolescence has negative effects on learning and memory that emerge by middle-age unless prevented by GH supplementation. PMID- 19815587 TI - Catch-up growth after dexamethasone withdrawal occurs in cultured postnatal rat metatarsal bones. AB - Children exposed to systemic glucocorticoids often exhibit growth retardation and after the cessation of therapy catch-up growth occurs in many, but not all patients. The developmental regulation and underlying cellular mechanisms of catch-up growth are not fully understood. To clarify this issue, we established an in vitro model of catch-up growth. Here we present a protocol for the long term culture (up to 160 days) of fetal (E20) as well as postnatal (P8) rat metatarsal bones which allowed us to characterize ex vivo the phenomenon of catch up growth without any influence by systemic factors. The relevance of the model was confirmed by the demonstration that the growth of fetal and postnatal bones were stimulated by IGF1 (100 ng/ml) and inhibited by dexamethasone (Dexa; 1 microM). We found that the capacity to undergo catch-up growth was restricted to postnatal bones. Catch-up growth occurred after postnatal bones had been exposed to Dexa for 7 or 12 days but not after a more prolonged exposure (19 days). Incomplete catch-up growth resulted in compromised bone length when assessed at the end of the 4-month period of culture. While exposure to Dexa was associated with decreased chondrocyte proliferation and differentiation, catch-up growth was only associated with increased cell proliferation. We conclude that the phenomenon of catch-up growth after Dexa treatment is intrinsic to the growth plate and primarily mediated by an upregulation of chondrocyte proliferation. PMID- 19815589 TI - Integrative Cancer Therapies. In this issue. PMID- 19815588 TI - NO-1886 suppresses diet-induced insulin resistance and cholesterol accumulation through STAT5-dependent upregulation of IGF1 and CYP7A1. AB - Insulin resistance and dyslipidemia are both considered to be risk factors for metabolic syndrome. Low levels of IGF1 are associated with insulin resistance. Elevation of low-density lipoprotein cholesterol (LDL-C) concomitant with depression of high-density lipoprotein cholesterol (HDL-C) increase the risk of obesity and type 2 diabetes mellitus (T2DM). Liver secretes IGF1 and catabolizes cholesterol regulated by the rate-limiting enzyme of bile acid synthesis from cholesterol 7alpha-hydroxylase (CYP7A1). NO-1886, a chemically synthesized lipoprotein lipase activator, suppresses diet-induced insulin resistance with the improvement of HDL-C. The goal of the present study is to evaluate whether NO 1886 upregulates IGF1 and CYP7A1 to benefit glucose and cholesterol metabolism. By using human hepatoma cell lines (HepG2 cells) as an in vitro model, we found that NO-1886 promoted IGF1 secretion and CYP7A1 expression through the activation of signal transducer and activator of transcription 5 (STAT5). Pretreatment of cells with AG 490, the inhibitor of STAT pathway, completely abolished NO-1886 induced IGF1 secretion and CYP7A1 expression. Studies performed in Chinese Bama minipigs pointed out an augmentation of plasma IGF1 elicited by a single dose administration of NO-1886. Long-term supplementation with NO-1886 recovered hyperinsulinemia and low plasma levels of IGF1 suppressed LDL-C and facilitated reverse cholesterol transport by decreasing hepatic cholesterol accumulation through increasing CYP7A1 expression in high-fat/high-sucrose/high-cholesterol diet minipigs. These findings indicate that NO-1886 upregulates IGF1 secretion and CYP7A1 expression to improve insulin resistance and hepatic cholesterol accumulation, which may represent an alternative therapeutic avenue of NO-1886 for T2DM and metabolic syndrome. PMID- 19815590 TI - Could integrative cancer treatment be cost-saving and resuscitate a submerged medical system? PMID- 19815591 TI - Drug-botanical interactions: a review of the laboratory, animal, and human data for 8 common botanicals. AB - Many Americans use complementary and alternative medicine (CAM) to prevent or alleviate common illnesses, and these medicines are commonly used by individuals with cancer.These medicines or botanicals share the same metabolic and transport proteins, including cytochrome P450 enzymes (CYP), glucuronosyltransferases (UGTs), and P-glycoprotein (Pgp), with over-the-counter and prescription medicines increasing the likelihood of drug-botanical interactions.This review provides a brief description of the different proteins, such as CYPs, UGTs, and Pgp.The potential effects of drug-botanical interactions on the pharmacokinetics and pharmacodynamics of the drug or botanical and a summary of the more common models used to study drug metabolism are described.The remaining portion of this review summarizes the data extracted from several laboratory, animal, and clinical studies that describe the metabolism, transport, and potential interactions of 8 selected botanicals. The 8 botanicals include black cohosh, Echinacea, garlic, Gingko biloba, green tea, kava, milk thistle, and St John's wort; these botanicals are among some of the more common botanicals taken by individuals with cancer.These examples are included to demonstrate how to interpret the different studies and how to use these data to predict the likelihood of a clinically significant drug-botanical interaction. PMID- 19815592 TI - A randomized controlled trial of the effects of transcendental meditation on quality of life in older breast cancer patients. AB - This single-blind, randomized controlled trial evaluated the impact of the Transcendental Meditation program plus standard care as compared with standard care alone on the quality of life (QOL) of older women (>or=55 years) with stage II to IV breast cancer. One hundred and thirty women (mean age = 63.8) were randomly assigned to either experimental (n = 64) or control (n = 66) groups. Functional Assessment of Cancer Therapy-Breast (FACT-B), Functional Assessment of Chronic Illness Therapy- Spiritual Well-Being (FACIT-SP), and Short-Form (SF)-36 mental health and vitality scales were administered every 6 months over an average 18-month intervention period. Significant improvements were found in the Transcendental Meditation group compared with controls in overall QOL, measured by the FACT-B total score (P = .037), emotional well-being (P = .046), and social well-being (P = .003) subscales, and SF-36 mental health ( P = .017). RESULTS: It is recommended that this stress reduction program, with its ease of implementation and home practice, be adopted in public health programs. PMID- 19815593 TI - Integrative Tumor Board: a case report and discussion from Dana-Farber Cancer Institute. AB - A 34-year-old woman carrying a BRCA1 gene and a significant family history was diagnosed with T1c, N1 breast cancer. The tumor was estrogen receptor, progesterone receptor, and HER-2/Neu negative. The patient received dose-dense chemotherapy with Adriamycin and Cytoxan followed by Taxol, and left breast irradiation. Later, a bilateral S-GAP flap reconstruction with right prophylactic mastectomy and left mastectomy were performed. During her treatment, the patient had an integrative medicine consultation and was seen by a team of health care providers specializing in integrative therapies, including integrative nutrition, therapeutic massage, acupuncture, and yoga. Each modality contributed unique benefit in her care that led to a satisfactory outcome for the patient. A detailed discussion regarding her care from each modality is presented. The case elucidates the need for integrative approaches for cancer patients in a conventional medical setting. PMID- 19815594 TI - Pomegranate fruit extract impairs invasion and motility in human breast cancer. AB - PURPOSE: Pomegranate fruit extracts (PFEs) possess polyphenolic and other compounds with antiproliferative, pro-apoptotic and anti-inflammatory effects in prostate, lung, and other cancers. Because nuclear transcription factor-kB (NF kB) is known to regulate cell survival, proliferation, tumorigenesis, and inflammation, it was postulated that PFEs may exert anticancer effects at least in part by modulating NF-kB activity. EXPERIMENTAL DESIGN: The authors investigated the effect of a novel, defined PFE consisting of both fermented juice and seed oil on the NF-kB pathway, which is constitutively active in aggressive breast cancer cell lines. The effects of the PFE on NF-kB-regulated cellular processes such as cell survival, proliferation, and invasion were also examined. RESULTS: Analytical characterization of the bioactive components of the PFE revealed active constituents, mainly ellagitannins and phenolic acids in the aqueous PFE and conjugated octadecatrienoic acids in the lipid PFE derived from seeds.The aqueous PFE dose-dependently inhibited NF-kB-dependent reporter gene expression associated with proliferation, invasion, and motility in aggressive breast cancer phenotypes while decreasing RhoC and RhoA protein expression. CONCLUSION: Inhibition of motility and invasion by PFEs, coincident with suppressed RhoC and RhoA protein expression, suggests a role for these defined extracts in lowering the metastatic potential of aggressive breast cancer species. PMID- 19815595 TI - Inhibition of viral carcinogenesis by Phyllanthus amarus. AB - Friend murine leukemia virus (FMuLv) is an acutely oncogenic retrovirus, and its infection leads to erythroblastosis and leukemia in mice. This infection model is used in the search for new antiviral agents. In the present study, the authors have evaluated the potential of an extract of Phyllanthus amarus against FMuLv induced erythroleukemia in BALB/c mice. Injection of newborn mice with FMuLv resulted in leukemia and animals died due to splenomegaly. Oral administration of P.amarus was found to enhance the life span of leukemia-harboring animals and decrease the incidence of anemia. The authors also performed a series of hematological, biochemical, histopathological, and gene expression analyses to evaluate the effect of P.amarus administration on erythroleukemia initiation and progression. The data obtained indicate that P.amarus administration could significantly decrease the progression of erythroleukemia. Treatment with P.amarus induced the expression of p53 and p45NFE2 and decreased the expression of Bcl-2 in the spleen of infected mice. Histopathological evaluations of the spleen demonstrated that administration of P.amarus decreased the infiltration of leukemic cells into the sinusoidal space when compared with the vehicle treated group. P.amarus is known to inhibit chemically induced neoplasm in different rodent models.The current results indicate that P.amarus has the ability to suppress virally induced cancers as well. PMID- 19815596 TI - Podophyllum hexandrum fraction (REC-2006) shows higher radioprotective efficacy in the p53-carrying hepatoma cell line: a role of cell cycle regulatory proteins. AB - The present study was carried out to evaluate the radioprotective efficacy of Podophyllum hexandrum fraction (REC-2006) in hepatoma cell lines having different p53 statuses. Higher radioresistance was observed in the HepG2 (p53(++)) cell line in comparison to the Hep3B (p53(-)) cell line, indicating a plausible role of p53 in radioresistance. REC-2006 exhibited nearly twice the survival in p53 expressing HepG2 cells compared with p53-negative Hep3B cells. REC-2006 treatment alone induced p53 expression as compared with untreated controls. However, REC 2006 reduced p53 expression when treated 2 hours before irradiation as compared with the irradiated HepG2 controls, indicating that REC-2006 modulates the expression of p53 to mitigate its apoptotic effect. Induction of p21 in the REC 2006 + radiation treatment group downregulated the expression of cyclin E and CDK2, leading to a delay in the G1 phase of HepG2 cells, which provided time for DNA repair or related processes. However, no significant difference in CDC2 expression in both cell lines suggested that G2 phase arrest might not be the only responsible factor for REC-2006-mediated radioprotection. Significant induction of PCNA and GADD45 expression in HepG2 cells suggested that REC-2006 increased the percentage survival of HepG2 cells by increasing the span of time as well as efficacy for repair processes. In conclusion, REC-2006 modulated the expression of p53 and thereby promoted cell cycle arrest in the G1 phase, encouraging cell proliferation and DNA repair and thus providing significantly higher protection against acute gamma-radiation in the HepG2 cell line. PMID- 19815597 TI - Anticancer activity of an Indian medicinal plant, Alstonia scholaris, on skin carcinogenesis in mice. AB - Alstonia scholaris, commonly known as sapthaparna, has been used for centuries in Ayurvedic medicine for treatment of various disorders. The objective of this study was to investigate the possible chemopreventive and anti-oxidative properties of this medicinal plant on two-stage process of skin carcinogenesis induced by a single application of 7, 12-dimethyabenz(a)anthrecene (100 lg/100 ll acetone), and two weeks later, promoted by repeated application of croton oil (1% in acetone/thrice a week) till the end of the experiment (16 weeks) in Swiss albino mice.The tumor incidence, tumor yield, tumor burden and cumulative number of papillomas were found to be higher in the carcinogen treated control (without ASE treatment) as compared to experimental animals (ASE treated). Furthermore, a significant increase in reduced glutathione, superoxide dismutase and catalase but decrease in lipid peroxidation was measured in ASE administered experimental groups than the carcinogen treated control. The present study demonstrates the chemopreventive potential of Alstonia scholaris bark extract in DMBA-induced skin tumorigenesis in Swiss albino mice. PMID- 19815598 TI - Gamma knife-treated hepatoma: case of obstructive jaundice and management. AB - Gamma knife therapy is becoming more frequently applied in solid tumor treatment. This article reports a unique case of severe obstructive jaundice arising as a complication of treatment of hepatoma at the hepatic hilum using a gamma knife.While planning an intervention, some images seem to promise success but actually lead to failure. Radiation damage to specific organs is difficult to predict because of several variables. Radiation-induced fibrosis and necrosis are the most common long-term adverse effects of radiotherapy; they are usually considered irreversible and result in induration and firmness of the tissue.To minimize radiation fibrosis, accurate patient positioning and tumor relocalization are essential for gamma knife use in the liver and other extracranial sites. Even when practiced frequently, any intervention must be delivered with caution if the liver has been treated with radiation. Otherwise, even with much experience, the unwary doctor can be trapped by deceptive images. PMID- 19815599 TI - Sonodynamic and photodynamic therapy in advanced breast carcinoma: a report of 3 cases. AB - Photodynamic therapy (PDT) is an established therapeutic method, first approved by the FDA for certain kinds of cancer in 1998. There are also increasing data to show that a related procedure, sonodynamic therapy (SDT), is a promising new modality for cancer treatment. Here, the authors report clinical results in 3 advanced refractory breast cancer patients who were treated using a combination of sonodynamic and photodynamic therapy (SPDT), along with conventional therapies. All 3 patients had pathologically proven metastatic breast carcinoma. These widely disseminated carcinomas had ultimately failed to respond to conventional therapy. A new sensitizing agent, Sonoflora 1 (SF1) was administered sublingually; then, after a 24-hour delay, patients were treated with a combination of light and ultrasound. All patients had significant partial or complete responses. SPDT is a promising new therapeutic combination for the treatment of breast cancer. PMID- 19815600 TI - Are positive alternative medical therapy trials credible?: Evidence from four high-impact medical journals. AB - Forty-five complementary and alternative medicine (CAM) efficacy randomized controlled trials (RCTs) from high-impact medical journals (NEJM, JAMA, Annals of Internal Medicine, and Archives of Internal Medicine) were reviewed based on their meeting three validity criteria (the existence of a placebo control, moderate attrition rates, and 50 or more participants per group). Of the 26 efficacy trials meeting all three criteria, only 2 (7.7%) were judged to be positive (i.e., the alternative therapy was significantly superior to its placebo control), while over half (55.5%) of the 19 trials that failed to meet one or more of these criteria reported positive results (p < .001). Of the two positive high-validity trials, one was funded and authored by the herbal company marketing the product tested and one used a placebo-control group of questionable credibility. This analysis is consistent with the hypothesis that CAM therapies are no more effective than placebos when adequate experimental control is present. PMID- 19815601 TI - Clinical trials of herbal treatments. AB - The deregulation of the marketing of dietary supplements with health claims has increased interest in evaluating the clinical effectiveness of these products. Clinical trials of herbal treatments pose challenges of limited preclinical data, lack of product standardization and characterization, and difficulties of blinding, which are substantially different from those in studies of conventional medications. These issues must be recognized and addressed if studies of herbal remedies are to provide useful information. PMID- 19815602 TI - Clinical features and management of arterial hypertension in children with Williams-Beuren syndrome. AB - BACKGROUND: Hypertension is a common finding in children with Williams-Beuren syndrome (WBS). METHODS: The aim of this retrospective study was to review the clinical presentation of systemic hypertension in WBS children, its origin and management. We included 41 children with confirmed WBS who were referred to the paediatric nephrology or cardiology unit for hypertension. RESULTS: The mean age at diagnosis of hypertension was 4.7 years. Out of 41, 24 patients had systolic blood pressure (BP) between +10 and +30 mmHg above the 95th percentile (1.645 SD), and 20/41 patients had diastolic BP between the 95th percentile (1.645 SD) and >10 mmHg. Thirty-nine patients were asymptomatic. Arteriography, performed in 17/41 patients, revealed a renal artery stenosis (RAS) in 10 patients (58%). Echocardiography was performed in all patients and showed isthmic coarctation in four patients (9%). Calcium channel blockers were used in half of the patients (22/41) and seemed to control hypertension in most cases. Interventional treatment of RAS was performed in five patients (three angioplasty and two surgical bypass). It controlled hypertension in one patient but remained ineffective in the four others. CONCLUSIONS: Medical treatment essentially calcium blockers improved hypertension in most cases. Interventional treatment of RAS has not been encouraging. PMID- 19815603 TI - Lack of association between the cholesteryl ester transfer protein gene--TaqIB polymorphism and coronary restenosis following percutaneous transluminal coronary angioplasty and stenting: a pilot study. AB - BACKGROUND: The most widely studied variation at the cholesteryl ester transfer protein (CETP) gene locus is a silent base change called the Thermobius aquaticus IB (TaqIB) polymorphism. TaqIB has been shown to affect levels/activity of CETP, plasma levels of high-density lipoprotein cholesterol (HDL-C), and to contribute to the risk of developing atherosclerosis and coronary heart disease (CHD). Ongoing studies are investigating possible associations between CETP gene polymorphisms and the development of coronary restenosis following percutaneous transluminal coronary angioplasty (PTCA) and stenting. METHODS AND RESULTS: The primary objective of the present study was to investigate the frequency of TaqIB polymorphism, and a possible association with post-PTCA coronary restenosis, in 204 Greek patients who had undergone PTCA and stenting. As a secondary objective, the analysis was extended to explore possible interacting or additive effects by various CHD risk factors, and a deletion in the alpha(2B)-adrenergic receptor gene. The frequency of TaqIB was 54%, similar to the frequency of the polymorphism in a group of 35 healthy controls. CONCLUSIONS: The results from this study do not indicate that the TaqIB variation at the CETP gene locus is a significant predictor for assessing the risk of developing coronary restenosis following PTCA and stenting. This result was not affected when considering any one of the additionally studied factors. PMID- 19815604 TI - The endothelium of basilar artery of diabetic rat treated with epoetin delta. AB - Erythropoiesis-stimulating agents (ESAs) are used to treat anemia associated with renal failure. It is now known that these agents also show a broad range of cell- and tissue-protective effects. In the current study, we explored whether an ESA, epoetin delta, affects vascular pathology linked to diabetes mellitus (DM). In a rat model of streptozotocin-induced DM, we investigated, by pre-embedding electron-immunocytochemistry, whether epoetin delta affects DM-induced structural changes in cerebrovascular endothelium of the rat basilar artery and influences the subcellular distribution of endothelial nitric oxide synthase (eNOS). Epoetin delta treatment influenced DM-induced changes to the distribution of eNOS in, and the structure of, the endothelial cell. This may indicate potential beneficial effects of epoetin delta on cerebrovascular endothelium and suggests eNOS as a possible target molecule of epoetin delta in DM. PMID- 19815605 TI - N-terminal ProBNP distribution and correlations with biological characteristics in apparently healthy Greek population: ATTICA study. AB - Brain natriuretic peptides are widely used as biomarkers of cardiovascular diseases and mainly heart failure. However, these markers are often found to be high even in apparently healthy participants, and little is known about which factors contribute to physiological change in plasma brain natriuretic peptide (BNP) and amino-terminal pro-B-type natriuretic peptide (NTproBNP) concentration in general populations. In this study, a random subsample of the ATTICA study was used (486 individuals) and serum NT-proBNP was measured. Approximately 20% of the participants had no detectable NT-proBNP values. Women had higher values of NT proBNP than men (median [25th-75th percentiles]: 30.2 [15.8-54.3] vs 14.9 [4.0 28.1] pg/mL, P < .001]. Amino-terminal pro-B-type natriuretic peptide values were positively correlated with age (rho = .140, P = .006) and inversely with body mass index (BMI; rho = -.142, P = .005), creatinine (Cr) clearance (rho = -.349, P < .001), and hemoglobin (rho = -.249, P < .001) values. Linear regression analysis revealed that gender is the main contributor of NT-proBNP levels, followed by age, BMI, and Cr values. PMID- 19815606 TI - Impact of transradial coronary procedures on radial artery. AB - OBJECTIVE: We evaluated the impact of transradial coronary procedures on the radial artery (RA). METHODS: A total of 355 patients who underwent the transradial coronary procedures including transradial coronary angiography (CAG) and percutaneous coronary intervention (PCI) were enrolled. The right RA (RRA) was examined by ultrasound. RESULTS: The mean RRA diameter was 2.37 +/- 0.57, 1.95 +/- 0.50, and 2.23 +/- 0.41 mm, respectively, before the procedure, 1 day, and 1 month after the procedures (P < .01 at 1 day, P < .05 at 1 month). The mean intima-media thickness of RRA was 0.25 +/- 0.12, 0.69 +/- 0.31, and 0.38 +/- 0.17 mm, respectively, before the procedure, 1 day and 1 month after the procedure (P < .01 at 1 day, P < .05 at 1 month). The incidence of RRA stenosis was 0%, 15.7%, and 7.6%, respectively; the incidence of RRA occlusion was 0%, 2.8%, and 1.7%, respectively, before the procedure,1 day and 1 month after the procedure. CONCLUSIONS: Transradial coronary procedures can lead to early RA injury, but this repairs later. PMID- 19815607 TI - Progress toward rapid and accurate Staphylococcus aureus strain typing. PMID- 19815608 TI - Quantitative transcription factor analysis of undifferentiated single human embryonic stem cells. AB - BACKGROUND: Human embryonic stem cells (hESCs) require expression of transcription factor genes POU5F1 (POU class 5 homeobox 1), NANOG (Nanog homeobox), and SOX2 [SRY (sex determining region Y)-box 2] to maintain their capacity for self-renewal and pluripotency. Because of the heterogeneous nature of cell populations, it is desirable to study the gene regulation in single cells. Large and potentially important fluctuations in a few cells cannot be detected at the population scale with microarrays or sequencing technologies. We used single-cell gene expression profiling to study cell heterogeneity in hESCs. METHODS: We collected 47 single hESCs from cell line SA121 manually by glass capillaries and 57 single hESCs from cell line HUES3 by flow cytometry. Single hESCs were lysed and reverse-transcribed. Reverse-transcription quantitative real time PCR was then used to measure the expression POU5F1, NANOG, SOX2, and the inhibitor of DNA binding genes ID1, ID2, and ID3. A quantitative noise model was used to remove measurement noise when pairwise correlations were estimated. RESULTS: The numbers of transcripts per cell varied >100-fold between cells and showed lognormal features. POU5F1 expression positively correlated with ID1 and ID3 expression (P < 0.05) but not with NANOG or SOX2 expression. When we accounted for measurement noise, SOX2 expression was also correlated with ID1, ID2, and NANOG expression (P < 0.05). CONCLUSIONS: We demonstrate an accurate method for transcription profiling of individual hESCs. Cell-to-cell variability is large and is at least partly nonrandom because we observed correlations between core transcription factors. High fluctuations in gene expression may explain why individual cells in a seemingly undifferentiated cell population have different susceptibilities for inductive cues. PMID- 19815609 TI - COLD-PCR-enhanced high-resolution melting enables rapid and selective identification of low-level unknown mutations. AB - BACKGROUND: Analysis of clinical samples often necessitates identification of low level somatic mutations within wild-type DNA; however, the selectivity and sensitivity of the methods are often limiting. COLD-PCR (coamplification at lower denaturation temperature-PCR) is a new form of PCR that enriches mutation containing amplicons to concentrations sufficient for direct sequencing; nevertheless, sequencing itself remains an expensive mutation-screening approach. Conversely, high-resolution melting (HRM) is a rapid, inexpensive scanning method, but it cannot specifically identify the detected mutation. To enable enrichment, quick scanning, and identification of low-level unknown mutations, we combined COLD-PCR with HRM mutation scanning, followed by sequencing of positive samples. METHODS: Mutation-containing cell-line DNA serially diluted into wild type DNA and DNA samples from human lung adenocarcinomas containing low-level mutations were amplified via COLD-PCR and via conventional PCR for TP53 (tumor protein p53) exons 6-8, and the 2 approaches were compared. HRM analysis was used to screen amplicons for mutations; mutation-positive amplicons were sequenced. RESULTS: Dilution experiments indicated an approximate 6- to 20-fold improvement in selectivity with COLD-PCR/HRM. Conventional PCR/HRM exhibited mutation detection limits of approximately 2% to 10%, whereas COLD-PCR/HRM exhibited limits from approximately 0.1% to 1% mutant-to-wild-type ratio. After HRM analysis of lung adenocarcinoma samples, we detected 7 mutations by both PCR methods in exon 7; however, in exon 8 we detected 9 mutations in COLD-PCR amplicons, compared with only 6 mutations in conventional-PCR amplicons. Furthermore, 94% of the HRM-detected mutations were successfully sequenced with COLD-PCR amplicons, compared with 50% with conventional-PCR amplicons. CONCLUSIONS: COLD-PCR/HRM improves the mutation-scanning capabilities of HRM and combines high selectivity, convenience, and low cost with the ability to sequence unknown low-level mutations in clinical samples. PMID- 19815610 TI - Increases of cardiac troponin in conditions other than acute coronary syndrome and heart failure. AB - BACKGROUND: Although cardiac troponin (cTn) is a cornerstone marker in the assessment and management of patients with acute coronary syndrome (ACS) and heart failure (HF), cTn is not diagnostically specific for any single myocardial disease process. This narrative review discusses increases in cTn that result from acute and chronic diseases, iatrogenic causes, and myocardial injury other than ACS and HF. CONTENT: Increased cTn concentrations have been reported in cardiac, vascular, and respiratory disease and in association with infectious processes. In cases involving acute aortic dissection, cerebrovascular accident, treatment in an intensive care unit, and upper gastrointestinal bleeding, increased cTn predicts a longer time to diagnosis and treatment, increased length of hospital stay, and increased mortality. cTn increases are diagnostically and prognostically useful in patients with cardiac inflammatory diseases and in patients with respiratory disease; in respiratory disease cTn can help identify patients who would benefit from aggressive management. In chronic renal failure patients the diagnostic sensitivity of cTn for ACS is decreased, but cTn is prognostic for the development of cardiovascular disease. cTn also provides useful information when increases are attributable to various iatrogenic causes and blunt chest trauma. SUMMARY: Information on the diagnostic and prognostic uses of cTn in conditions other than ACS and heart failure is accumulating. Although increased cTn in settings other than ACS or heart failure is frequently considered a clinical confounder, the astute physician must be able to interpret cTn as a dynamic marker of myocardial damage, using clinical acumen to determine the source and significance of any reported cTn increase. PMID- 19815611 TI - Staphylococcus aureus strain typing by single-molecule DNA mapping in fluidic microchips with fluorescent tags. AB - BACKGROUND: Epidemiologic studies require identification or typing of microbial strains. Macrorestriction DNA mapping analyzed by pulsed-field gel electrophoresis (PFGE) is considered the current gold standard of genomic typing. This technique, however, is difficult to implement because it is labor-intensive and difficult to automate, it requires a long time to obtain results, and results often vary between laboratories. METHODS: We used direct linear analysis (DLA), which uses a single reagent set and long fragments of microbial genomic DNA to identify various microbes. In this technique, an automated system extracts fragments exceeding 100 kb from restriction enzyme digests of genomic DNA from microbial isolates and hybridizes them with a sequence-dependent fluorescent tag. These fragments are then stretched in a microfluidics chip, and the patterns of the distribution of the tags are discerned with fluorescence confocal microscopy. The tag pattern on each DNA fragment is compared with a database of known microbial DNA sequences or with measured patterns of other microbial DNAs. RESULTS: We used DLA to type 71 Staphylococcus aureus strains. Of these, 9 had been sequenced, 10 were representative of the major pulsed-field types present in the US, and 52 were isolated recently in a hospital in Cambridge, MA. Matching DNA fragments were identified in different samples by a clustering algorithm and were used to quantify the similarities of the strains. CONCLUSIONS: DLA-based strain typing is a powerful technique with a resolution comparable to macrorestriction mapping with PFGE, but DLA is faster, more automated, and more reproducible. PMID- 19815612 TI - Delayed onset of acute renal failure after significant paracetamol overdose: A case series. AB - Acute renal failure is a recognized manifestation of paracetamol toxicity, but comparatively little data is available concerning its onset and duration. The present study sought to characterize the time course of rising serum creatinine concentrations in paracetamol nephrotoxicity. Renal failure was defined by serum creatinine concentration >or=150 micromol/L (1.69 mg/dL) or >or=50% increase from baseline. Serum creatinine concentrations and alanine aminotransferase activity were considered with respect to the interval after paracetamol ingestion. There were 2068 patients with paracetamol overdose between March 2005 and October 2007, and paracetamol nephrotoxicity occurred in 8 (0.4%). All had significant hepatotoxicity, and peak serum alanine aminotransferase activity occurred at 2.5 days (2.2 to 2.9 days) after ingestion. Peak serum creatinine concentrations did not occur until 5.5 days (4.4 to 5.9 days) after ingestion (p = .031 by Wilcoxon test). Serum creatinine concentrations slowly restored to normal, and renal replacement was not required. In this patient series, rising serum creatinine concentrations only became detectable after more than 48 hours after paracetamol ingestion. Therefore, renal failure might easily be missed if patients are discharged home before this. Further work is required to establish the prevalence of paracetamol-induced nephrotoxicity, and its clinical significance. PMID- 19815613 TI - Ultrastructural disorder of the secretory pathway in temperature-sensitive actin mutants of Saccharomyces cerevisiae. AB - Phenotypes of the two temperature-sensitive actin mutants of Saccharomyces cerevisiae act1-1 and act1-2 at permissive, restrictive and semi-restrictive temperatures were studied by freeze fracture and thin section electron microscopy, and fluorescent microscopy. In contrast to secretory mutants where accumulations of either secretory vesicles, Golgi apparatus, or endoplasmic reticulum were reported, act1-1 and act1-2 mutants revealed accumulation of all the three components, even at permissive temperature. However, more distinct accumulation of secretory organelles was evident during cultivation at the sub restrictive temperature of 30 degrees C. At the restrictive temperature of 37 degrees C, many cells died, and their empty cell walls remained. Some of the few living cells showed features of apoptosis. From the present study, actin cables are concluded to be necessary for (i) correct spatial positioning and orientation of secretary pathway to the bud and septum, and (ii) vectorial movement of vesicles of the secretory pathway along the actin cables to the bud and septum. PMID- 19815614 TI - Impact of front-of-pack 'traffic-light' nutrition labelling on consumer food purchases in the UK. AB - Front-of-pack 'traffic-light' nutrition labelling has been widely proposed as a tool to improve public health nutrition. This study examined changes to consumer food purchases after the introduction of traffic-light labels with the aim of assessing the impact of the labels on the 'healthiness' of foods purchased. The study examined sales data from a major UK retailer in 2007. We analysed products in two categories ('ready meals' and sandwiches), investigating the percentage change in sales 4 weeks before and after traffic-light labels were introduced, and taking into account seasonality, product promotions and product life-cycle. We investigated whether changes in sales were related to the healthiness of products. All products that were not new and not on promotion immediately before or after the introduction of traffic-light labels were selected for the analysis (n = 6 for ready meals and n = 12 for sandwiches). For the selected ready-meals, sales increased (by 2.4% of category sales) in the 4 weeks after the introduction of traffic-light labels, whereas sales of the selected sandwiches did not change significantly. Critically, there was no association between changes in product sales and the healthiness of the products. This short-term study based on a small number of ready meals and sandwiches found that the introduction of a system of four traffic-light labels had no discernable effect on the relative healthiness of consumer purchases. Further research on the influence of nutrition signposting will be needed before this labelling format can be considered a promising public health intervention. PMID- 19815615 TI - T2* signal hyperintensity in subacute cerebral vein thrombosis. PMID- 19815616 TI - Randomized vertebroplasty trials: bad news or sham news? PMID- 19815617 TI - Response to "randomized vertebroplasty trials: bad news or sham news?". PMID- 19815618 TI - MR imaging findings in brachial plexopathy with thoracic outlet syndrome. AB - The BPL is a part of the peripheral nervous system. Many disease processes affect the BPL. In this article, on the basis of 60 patients, we reviewed MR imaging findings of subjects with brachial plexopathy. Different varieties of BPL lesions are discussed. PMID- 19815619 TI - Human retinopathy-associated ciliary protein retinitis pigmentosa GTPase regulator mediates cilia-dependent vertebrate development. AB - Dysfunction of primary cilia is associated with tissue-specific or syndromic disorders. RPGR is a ciliary protein, mutations in which can lead to retinitis pigmentosa (RP), cone-rod degeneration, respiratory infections and hearing disorders. Though RPGR is implicated in ciliary transport, the pathogenicity of RPGR mutations and the mechanism of underlying phenotypic heterogeneity are still unclear. Here we have utilized genetic rescue studies in zebrafish to elucidate the effect of human disease-associated mutations on its function. We show that rpgr is expressed predominantly in the retina, brain and gut of zebrafish. In the retina, RPGR primarily localizes to the sensory cilium of photoreceptors. Antisense morpholino (MO)-mediated knockdown of rpgr function in zebrafish results in reduced length of Kupffer's vesicle (KV) cilia and is associated with ciliary anomalies including shortened body-axis, kinked tail, hydrocephaly and edema but does not affect retinal development. These phenotypes can be rescued by wild-type (WT) human RPGR. Several of the RPGR mutants can also reverse the MO induced phenotype, suggesting their potential hypomorphic function. Notably, selected RPGR mutations observed in XLRP (T99N, E589X) or syndromic RP (T124fs, K190fs and L280fs) do not completely rescue the rpgr-MO phenotype, indicating a more deleterious effect of the mutation on the function of RPGR. We propose that RPGR is involved in cilia-dependent cascades during development in zebrafish. Our studies provide evidence for a heterogenic effect of the disease-causing mutations on the function of RPGR. PMID- 19815621 TI - Brain anatomy changes associated with persistent neuropathic pain following spinal cord injury. AB - Persistent neuropathic pain commonly occurs following spinal cord injury (SCI). It remains one of the most challenging management problems in this condition. In order to develop more effective treatments, a better understanding of the neural changes associated with neuropathic SCI pain is required. The aim of this investigation was to use diffusion tensor imaging (DTI) to determine if persistent neuropathic pain following SCI is associated with changes in regional brain anatomy and connectivity. In 23 subjects with complete thoracic SCI, 12 with below-level neuropathic pain and 11 without pain, and 45 healthy control subjects, a series of whole-brain DTI scans were performed. The mean diffusivity (MD) of each voxel was calculated and values compared between groups. This analysis revealed that neuropathic pain following SCI is associated with significant differences in regional brain anatomy. These anatomical changes were located in pain-related regions as well as regions of the classic reward circuitry, that is, the nucleus accumbens and orbitofrontal, dorsolateral prefrontal, and posterior parietal cortices. The right posterior parietal cortex projected to most regions that displayed an anatomical change. Analysis of the fiber tracts connecting areas of MD differences revealed no significance differences in MD values between the SCI pain, SCI no pain, and control groups. PMID- 19815620 TI - Impaired replication dynamics at the FRA3B common fragile site. AB - Chromosomal common fragile sites (CFSs) are genetically unstable regions of the genome that are induced by conditions that impair DNA replication. In this report, we show that treatment with the DNA polymerase inhibitor, aphidicolin (APH), slows the replication rate throughout S phase. To investigate the unusual sensitivity of CFSs to APH-induced replication stress, we examined replication dynamics within a 50 kb region of the most frequently expressed CFS, FRA3B. We mapped four origins of replication, ori 1-4, using two independent methods. In untreated cells, we detected significantly less newly replicated DNA at FRA3B ori 1-3, as compared with three control origins located within non-fragile regions (NCFSs). In APH-treated cells, all FRA3B and control origins tested were active; however, there was a significant increase of nascent strand DNA at the control origins and, to a lesser extent, at the FRA3B ori 1-3. On the basis of these observations and the theoretical modeling of the nascent strand abundance assay developed in this study, we hypothesize that CFS origins may be less efficient, and that APH treatment slows replication fork movement near these origins to a greater extent, resulting in impaired DNA replication and, ultimately, leading to the genetic instability characteristic of CFSs. PMID- 19815622 TI - Embryonic stem cell-like cells derived from adult human testis. AB - BACKGROUND: Given the significant drawbacks of using human embryonic stem (hES) cells for regenerative medicine, the search for alternative sources of multipotent cells is ongoing. Studies in mice have shown that multipotent ES-like cells can be derived from neonatal and adult testis. Here we report the derivation of ES-like cells from adult human testis. METHODS: Testis material was donated for research by four men undergoing bilateral castration as part of prostate cancer treatment. Testicular cells were cultured using StemPro medium. Colonies that appeared sharp edged and compact were collected and subcultured under hES-specific conditions. Molecular characterization of these colonies was performed using RT-PCR and immunohistochemistry. (Epi)genetic stability was tested using bisulphite sequencing and karyotype analysis. Directed differentiation protocols in vitro were performed to investigate the potency of these cells and the cells were injected into immunocompromised mice to investigate their tumorigenicity. RESULTS: In testicular cell cultures from all four men, sharp-edged and compact colonies appeared between 3 and 8 weeks. Subcultured cells from these colonies showed alkaline phosphatase activity and expressed hES cell-specific genes (Pou5f1, Sox2, Cripto1, Dnmt3b), proteins and carbohydrate antigens (POU5F1, NANOG, SOX2 and TRA-1-60, TRA-1-81, SSEA4). These ES-like cells were able to differentiate in vitro into derivatives of all three germ layers including neural, epithelial, osteogenic, myogenic, adipocyte and pancreatic lineages. The pancreatic beta cells were able to produce insulin in response to glucose and osteogenic-differentiated cells showed deposition of phosphate and calcium, demonstrating their functional capacity. Although we observed small areas with differentiated cell types of human origin, we never observed extensive teratomas upon injection of testis-derived ES-like cells into immunocompromised mice. CONCLUSIONS: Multipotent cells can be established from adult human testis. Their easy accessibility and ethical acceptability as well as their non-tumorigenic and autogenic nature make these cells an attractive alternative to human ES cells for future stem cell therapies. PMID- 19815623 TI - Hepatic phenotype of liver fatty acid binding protein gene-ablated mice. AB - Although the function of liver fatty acid binding protein in hepatic fatty acid metabolism has been extensively studied, its potential role in hepatic cholesterol homeostasis is less clear. Although hepatic cholesterol accumulation was initially reported in L-FABP-null female mice, that study was performed with early N2 backcross generation mice. To resolve whether the hepatic cholesterol phenotype in these L-FABP(-/-) mice was attributable to genetic inhomogeneity, these L-FABP(-/-) mice were further backcrossed to C57Bl/6 mice up to the N10 (99.9% homogeneity) generation. Hepatic total cholesterol accumulation was observed in female, but not male, L-FABP(-/-) mice at all (N2, N4, N6, N10) backcross generations examined. The greater total cholesterol was due to increased hepatic levels of both unesterified (free) cholesterol and esterified cholesterol. Altered hepatic cholesterol accumulation correlated directly with L FABP's ability to bind cholesterol with high affinity as shown by direct L-FABP binding of fluorescent cholesterol analogs (NBD-cholesterol, dansyl-cholesterol), a photoactivatable cholesterol analog [free cholesterol benzophenone (FCBP)], and free cholesterol (circular dichroism, isothermal titration microcalorimetry). One mole of fluorescent sterol was bound per mole of L-FABP. This was confirmed by photo-cross-linking studies with the photoactivatable cholesterol analog FCBP and by isothermal titration calorimetry with free cholesterol, which showed that L FABP bound only one sterol molecule per L-FABP molecule. In contrast, the hepatic phenotype of male, but not female, L-FABP(-/-) mice was characterized by decreased hepatic triacylglycerol levels at all backcross generations examined. Taken together, these data support the hypothesis that L-FABP plays a role in physiological regulation of not only hepatic fatty acid metabolism, but also that of hepatic cholesterol. PMID- 19815624 TI - Mediators released from LPS-challenged lungs induce inflammatory responses in liver vascular endothelial cells and neutrophilic leukocytes. AB - The systemic inflammatory response plays an important role in the progression of acute lung injury (ALI) to multiple organ dysfunction syndrome (MODS). However, the role of lung-derived inflammatory mediators in induction of the inflammatory response in remote organs is poorly understood. To address the above, we investigated the effects of lung inflammation on induction of inflammatory response(s) in the liver in vitro. Inflammation in mouse lungs was induced by intranasal administration of lipopolysaccharide (LPS; 1 mg/ml) followed by mechanical ventilation using the isolated perfused mouse lung method to obtain and characterize lung perfusate from the pulmonary circulation. LPS administration to mouse lungs resulted in an increased release of inflammation relevant cytokines and chemokines into the perfusate (Luminex assay) compared with the saline-controls. Subsequently, primary mouse liver vascular endothelial cells (LVEC) or mouse polymorphonuclear leukocytes (PMN) in vitro were stimulated with the perfusate obtained from saline- or LPS-challenged lungs and assessed for various inflammation-relevant end points. The obtained results indicate that stimulation of LVEC with perfusate obtained from LPS-challenged lungs results in 1) reactive oxygen species (ROS) production; 2) activation of NF-kappaB; and 3) expression of E-selectin, ICAM-1, and VCAM-1 and a subsequent increase in PMN rolling and adhesion to LVEC. In addition, perfusate from LPS-challenged lung induced activation of PMN with respect to increased ROS production and upregulation of cell surface levels of adhesion molecules MAC-1 and VLA-4. Heat inactivation of the perfusate obtained from LPS-challenged lungs was very effective in suppressing increased proadhesive phenotype (i.e., E-selectin and ICAM-1 expression) in LVEC, whereas targeted inhibition (immunoneutralization) of TNF-alpha and/or IL-6 in LPS-lung perfusate had no effect. Taken together, these findings indicate that multiple proinflammatory mediators (proteinaceous in nature) released from inflamed lungs act synergistically to induce systemic activation of circulating PMN and promote inflammatory responses in liver vascular endothelial cells. PMID- 19815625 TI - PKC{epsilon}-dependent and -independent effects of taurolithocholate on PI3K/PKB pathway and taurocholate uptake in HuH-NTCP cell line. AB - The cholestatic bile acid taurolithocholate (TLC) inhibits biliary secretion of organic anions and hepatic uptake of taurocholate (TC). TLC has been suggested to induce retrieval of Mrp2 from the canalicular membrane via the phosphoinositide-3 kinase (PI3K)/PKB-dependent activation of novel protein kinase Cepsilon (nPKCepsilon) in rat hepatocytes. The aim of the present study was to determine whether TLC-induced inhibition of TC uptake may also involve PI3K-dependent activation of PKCepsilon in HuH7 cells stably transfected with human Na(+) dependent TC-cotransporting polypeptide (NTCP) (HuH-NTCP cells). To avoid direct competition for uptake, cells were pretreated with TLC, washed, and then incubated with (3)H-TC to determine TC uptake. TLC produced time- and dose dependent inhibition of TC uptake. TLC inhibited TC uptake competitively without affecting NTCP membrane translocation. A PI3K inhibitor failed to reverse TLC induced TC uptake inhibition and TLC-inhibited PKB phosphorylation. TLC did activate nPKCepsilon as evidenced by increased membrane translocation and nPKCepsilon-Ser(729) phosphorylation. Overexpression of dominant negative nPKCepsilon reversed TLC-induced inhibition of PKB phosphorylation but not of TC uptake. Finally, cAMP prevented TLC-induced inhibition of TC uptake via the PI3K pathway, and the prevention is due to the sum of cAMP-induced stimulation and TLC induced inhibition of TC uptake. Taken together, these results suggest that TLC induced inhibition of PKB, but not of TC uptake, is mediated via nPKCepsilon. Activation of nPKCepsilon and inhibition of TC uptake by TLC are not mediated via the PI3K/PKB pathway. PMID- 19815626 TI - Cholecystokinin-58 and cholecystokinin-8 exhibit similar actions on calcium signaling, zymogen secretion, and cell fate in murine pancreatic acinar cells. AB - The gastrointestinal hormone CCK exists in various molecular forms, with differences in bioactivity between the well-characterized CCK-8 and larger CCK-58 previously reported. We have compared the effects of these peptides on cytosolic calcium concentration ([Ca(2+)](c)), mitochondrial metabolism, enzyme secretion, and cell fate in murine isolated pancreatic acinar cells using fluorescence confocal microscopy and patch-clamp electrophysiology. CCK-58 (1-10 pM) induced transient, oscillatory increases of [Ca(2+)](c), which showed apical to basolateral progression and were associated with a rise of mitochondrial NAD(P)H. CCK-58 (10 pM) induced zymogen exocytosis in isolated cells and amylase secretion from isolated cells and whole tissues. Hyperstimulation with supraphysiological CCK-58 (5 nM) induced a single large increase of [Ca(2+)](c) that declined to a plateau, which remained above the basal level 20 min after application and was dependent on external Ca(2+) entry. In cells dispersed from the same tissues, CCK 8 induced similar patterns of responses to those of CCK-58, with oscillatory increases of [Ca(2+)](c) at lower (pM) concentrations and sustained responses at 5 nM. CCK-58 and CCK-8 exhibited similar profiles of action on cell death, with increases in necrosis at high CCK-58 and CCK-8 (10 nM) that were not significantly different between peptides. The present experiments indicate that CCK-8 and CCK-58 have essentially identical actions on the acinar cell at high and low agonist concentrations, suggesting an action via the same receptor and that the differences observed in an intact rat model may result from indirect effects of the peptides. Our data strengthen the argument that CCK-58 is an important physiological form of this gastrointestinal hormone. PMID- 19815627 TI - Cilostazol, a specific PDE-3 inhibitor, ameliorates chronic ileitis via suppression of interaction of platelets with monocytes. AB - Excessive migration of monocytes to a site of intestinal inflammation contributes to tissue damage in Crohn's disease. It is known that cilostazol, a specific phosphodiesterase-3 (PDE-3) inhibitor of platelets, decreases monocyte recruitment to intestinal mucosa through suppression of platelet-monocyte interactions. The objective of this study was to clarify whether cilostazol ameliorates murine ileitis by suppression of monocyte migration. Significant inflammation was induced in the ileum of SAMP1/Yit mice at 23 wk of age after piroxicam treatment for 3 wk. Weight of the terminal ileum of mice was significantly greater with inflammatory cell infiltration in SAMP1/Yit mice than in control mice (AKR-J). Treatment of SAMP1/Yit mice with cilostazol-containing food (200 ppm) for 3 wk significantly attenuated the increase in intestinal weight and the histological changes, including invasion of F4/80-positive macrophages. A significant increase in migration of monocytes and platelets to microvessels of the ileal mucosa was observed in SAMP/Yit mice in vivo by using an intravital fluorescence microscope. Pretreatment with cilostazol significantly attenuated the increased migration of monocytes, possibly through suppression of platelet-monocyte interactions. In conclusion, a PDE-3 inhibitor ameliorates murine ileitis through attenuating migration of monocytes to the intestinal mucosa, suggesting a potential usefulness of antiplatelet drugs for treatment of Crohn's disease. PMID- 19815628 TI - Hedgehog pathway activation and epithelial-to-mesenchymal transitions during myofibroblastic transformation of rat hepatic cells in culture and cirrhosis. AB - Myofibroblastic hepatic stellate cells (MF-HSC) are derived from quiescent hepatic stellate cells (Q-HSC). Q-HSC express certain epithelial cell markers and have been reported to form junctional complexes similar to epithelial cells. We have shown that Hedgehog (Hh) signaling plays a key role in HSC growth. Because Hh ligands regulate epithelial-to-mesenchymal transition (EMT), we determined whether Q-HSC express EMT markers and then assessed whether these markers change as Q-HSC transition into MF-HSC and whether the process is modulated by Hh signaling. Q-HSC were isolated from healthy livers and cultured to promote myofibroblastic transition. Changes in mRNA and protein expression of epithelial and mesenchymal markers, Hh ligands, and target genes were monitored in HSC treated with and without cyclopamine (an Hh inhibitor). Studies were repeated in primary human HSC and clonally derived HSC from a cirrhotic rat. Q-HSC activation in vitro (culture) and in vivo (CCl(4)-induced cirrhosis) resulted in decreased expression of Hh-interacting protein (Hhip, an Hh antagonist), the EMT inhibitors bone morphogenic protein (BMP-7) and inhibitor of differentiation (Id2), the adherens junction component E-cadherin, and epithelial keratins 7 and 19 and increased expression of Gli2 (an Hh target gene) and mesenchymal markers, including the mesenchyme-associated transcription factors Lhx2 and Msx2, the myofibroblast marker alpha-smooth muscle actin, and matrix molecules such as collagen. Cyclopamine reverted myofibroblastic transition, reducing mesenchymal gene expression while increasing epithelial markers in rodent and human HSC. We conclude that Hh signaling plays a key role in transition of Q-HSC into MF-HSC. Our findings suggest that Q-HSC are capable of transitioning between epithelial and mesenchymal fates. PMID- 19815629 TI - The normal mechanisms of pregnancy-induced liver growth are not maintained in mice lacking the bile acid sensor Fxr. AB - Rodents undergo gestational hepatomegaly to meet the increased metabolic demands on the maternal liver during pregnancy. This is an important physiological process, but the mechanisms and signals driving pregnancy-induced liver growth are not known. Here, we show that liver growth during pregnancy precedes maternal body weight gain, is proportional to fetal number, and is a result of hepatocyte hypertrophy associated with cell-cycle progression, polyploidy, and altered expression of cell-cycle regulators p53, Cyclin-D1, and p27. Because circulating reproductive hormones and bile acids are raised in normal pregnant women and can cause liver growth in rodents, these compounds are candidates for the signal driving gestational liver enlargement in rodents. Administration of pregnancy levels of reproductive hormones was not sufficient to cause liver growth, but mouse pregnancy was associated with increased serum bile acid levels. It is known that the bile acid sensor Fxr is required for normal recovery from partial hepatectomy, and we demonstrate that Fxr(-/-) mice undergo gestational liver growth by adaptive hepatocyte hyperplasia. This is the first identification of any component that is required to maintain the normal mechanisms of gestational hepatomegaly and also implicates Fxr in a physiologically normal process that involves control of the hepatocyte cell cycle. Understanding pregnancy-induced hepatocyte hypertrophy in mice could suggest mechanisms for safely increasing functional liver capacity in women during increased metabolic demand. PMID- 19815630 TI - Characterizing the application of transcranial direct current stimulation in human pharyngeal motor cortex. AB - Transcranial direct current stimulation (tDCS) is a novel intervention that can modulate brain excitability in health and disease; however, little is known about its effects on bilaterally innervated systems such as pharyngeal motor cortex. Here, we assess the effects of differing doses of tDCS on the physiology of healthy human pharyngeal motor cortex as a prelude to designing a therapeutic intervention in dysphagic patients. Healthy subjects (n = 17) underwent seven regimens of tDCS (anodal 10 min 1 mA, cathodal 10 min 1 mA, anodal 10 min 1.5 mA, cathodal 10 min 1.5 mA, anodal 20 min 1 mA, cathodal 20 min 1 mA, Sham) on separate days, in a double blind randomized order. Bihemispheric motor evoked potential (MEP) responses to single-pulse transcranial magnetic stimulation (TMS) as well as intracortical facilitation (ICF) and inhibition (ICI) were recorded using a swallowed pharyngeal catheter before and up to 60 min following the tDCS. Compared with sham, both 10 min 1.5 mA and 20 min 1 mA anodal stimulation induced increases in cortical excitability in the stimulated hemisphere (+44 +/- 17% and +59 +/- 16%, respectively; P < 0.005) whereas only 10 min 1.5 mA cathodal stimulation induced inhibition (-26 +/- 4%, P = 0.02). There were neither contralateral hemisphere changes nor any evidence for ICI or ICF in driving the ipsilateral effects. In conclusion, anodal tDCS can alter pharyngeal motor cortex excitability in an intensity-dependent manner, with little evidence for transcallosal spread. Anodal stimulation may therefore provide a useful means of stimulating pharyngeal cortex and promoting recovery in dysphagic patients. PMID- 19815631 TI - Prenatal flavor exposure affects flavor recognition and stress-related behavior of piglets. AB - Exposure to flavors in the amniotic fluid and mother's milk derived from the maternal diet has been shown to modulate food preferences and neophobia of young animals of several species. Aim of the experiment was to study the effects of pre and postnatal flavor exposure on behavior of piglets during (re)exposure to this flavor. Furthermore, we investigated whether varying stress levels, caused by different test settings, affected behavior of animals during (re)exposure. Piglets were exposed to anisic flavor through the maternal diet during late gestation and/or during lactation or never. Piglets that were prenatally exposed to the flavor through the maternal diet behaved differently compared with unexposed pigs during reexposure to the flavor in several tests, suggesting recognition of the flavor. The differences between groups were more pronounced in tests with relatively high stress levels. This suggests that stress levels, caused by the design of the test, can affect the behavior shown in the presence of the flavor. We conclude that prenatal flavor exposure affects behaviors of piglets that are indicative of recognition and that these behaviors are influenced by stress levels during (re)exposure. PMID- 19815632 TI - Augmented effect of early antibiotic treatment in mice with experimental lung infections due to sequentially adapted mucoid strains of Pseudomonas aeruginosa. AB - BACKGROUND: Effects of treatment with tobramycin initiated 1 or 24 h post infection were investigated in a new version of a pulmonary infection model in mice. The model reflects the differentiated behaviour of Pseudomonas aeruginosa mucoid strains isolated from the lungs of one chronically infected cystic fibrosis (CF) patient at different time periods during chronic lung infection. METHODS: BALB/c mice were challenged with alginate-embedded mucoid clinical isolates isolated in 1988, 1997 or 2003. Mice were euthanized on day 1, 2 or 3 post-infection for estimation of quantitative bacteriology, histopathology, and measurement of granulocyte colony-stimulating factor (G-CSF) and macrophage inflammatory protein 2 (MIP-2). RESULTS: There was a significant reduction of bacteria when comparing treatment initiated 1 h post-infection with treatment initiated after 24 h for isolates 1997 and 2003. Treatment initiated 1 h post infection also resulted in a reduction of the pulmonary cytokines G-CSF, for all three isolates, and MIP-2, for isolates 1997 and 2003. Histological evaluation showed a shift from the acute-type inflammatory immune response to a chronic-type in mice infected with isolate 2003. CONCLUSIONS: A significant reduction in the number of bacteria was observed when initiating treatment 1 h post-infection compared with initiating treatment after 24 h, although the latest isolate avoided complete clearance. Early antibiotic treatment directed at the mucoid phenotype in mice also reduced the inflammation and, thereby, the lung tissue damage. PMID- 19815633 TI - Emergence of extended-spectrum beta-lactamases and AmpC-type beta-lactamases in human Salmonella isolated in Spain from 2001 to 2005. AB - OBJECTIVES: To study the resistance to third-generation cephalosporins in Salmonella strains isolated from humans in a 5 year period in Spain, and to identify the responsible genes and their dissemination. METHODS: Twenty-seven isolates were analysed by PCR and sequencing to identify the genes responsible for the beta-lactamase resistance phenotypes. The transferability of the phenotypes was tested by conjugation to Escherichia coli K12J53, plasmid detection with S1-PFGE, hybridization and PCRs of the transconjugants. The genetic relationship was determined by PFGE. RESULTS: We found bla(CTX-M-9) and bla(CTX-M-10) in Salmonella Virchow PT19. bla(CTX-M-14) was detected in Salmonella (IV) 44:z(4),z(23):-, Salmonella Enteritidis PT6a, Salmonella Typhimurium DT193 and Salmonella Typhimurium DT104B. bla(CTX-M-1) was found in Salmonella Litchfield. bla(CTX-M-15) and bla(CTX-M-32) were found in Salmonella Enteritidis PT1. bla(SHV-12) was found in Salmonella Blockley, Salmonella Hadar PT2, Salmonella Enteritidis PT21, Salmonella Enteritidis PT1 and Salmonella Bredeney. bla(SHV-2) was found in Salmonella Livingstone. bla(CMY-2) was detected in Salmonella Bredeney, Salmonella Newport, Salmonella Enteritidis PT5b and Salmonella Heidelberg. bla(DHA-1) was detected for the first time in Spain in Salmonella Newport. One strain of Salmonella Senftenberg harboured two extended spectrum beta-lactamases, bla(SHV-12) and bla(CTX-M-9). We have found a large variety of beta-lactamase families as well as several members of major relevance, such as CTX-M-15, CTX-M-32, CMY-2 and DHA-1. XbaI-PFGE, conjugation assays and S1 PFGE hybridization showed that all these beta-lactamases were mediated by plasmids. CONCLUSIONS: This study demonstrates the emergence of a public health risk related to resistance to beta-lactams in Salmonella. The resistance trends need to be monitored carefully. PMID- 19815635 TI - Moderator effects in a randomized controlled trial of exercise training in lymphoma patients. AB - BACKGROUND: The Healthy Exercise for Lymphoma Patients trial showed that aerobic exercise training improved important health outcomes in lymphoma patients. Here, we examine potential moderators of the exercise training response. METHODS: Lymphoma patients were stratified by major disease type and current treatment status and randomly assigned to usual care (n = 62) or aerobic exercise training (n = 60) for 12 weeks. Endpoints were quality of life, cardiovascular fitness, and body composition. Moderators were patient preference for group assignment, age, sex, marital status, disease stage, body mass index, and general health. RESULTS: Patient preference did not statistically moderate the effects of exercise training on quality of life (P for interaction = 0.36), but the interaction effect of 7.8 points favoring patients with no preference was clinically meaningful. Marital status (P for interaction = 0.083), general health (P for interaction = 0.012), and body mass index (P for interaction = 0.010) moderated the effects of aerobic exercise training on quality of life with better outcomes for unmarried versus married patients, patients in poor/fair health versus good-to-excellent health, and normal weight/obese versus overweight patients. Disease stage (P for interaction = 0.056) and general health (P for interaction = 0.012) moderated the effects of aerobic exercise training on body composition with better outcomes for patients with advanced disease versus early disease/no disease and patients in good health versus very good-to-excellent health. No variables moderated intervention effects on cardiovascular fitness. Findings were not explained by differences in adherence. CONCLUSIONS: Clinically available variables predicted quality of life and body composition responses to aerobic exercise training in lymphoma patients. If replicated, these results may inform future randomized trials and clinical practice. PMID- 19815634 TI - Association of area socioeconomic status and breast, cervical, and colorectal cancer screening: a systematic review. AB - BACKGROUND: Although numerous studies have examined the association of area socioeconomic status (SES) and cancer screening after controlling for individual SES, findings have been inconsistent. A systematic review of existing studies is timely to identify conceptual and methodologic limitations and to provide a basis for future research directions and policy. OBJECTIVE: The objectives were to (a) describe the study designs, constructs, methods, and measures; (b) describe the independent association of area SES and cancer screening; and (c) identify neglected areas of research. METHODS: We searched six electronic databases and manually searched cited and citing articles. Eligible studies were published before 2008 in peer-reviewed journals in English, represented primary data on individuals ages > or = 18 years from developed countries, and measured the association of area and individual SES with breast, cervical, or colorectal cancer screening. RESULTS: Of 19 eligible studies, most measured breast cancer screening. Studies varied widely in research design, definitions, and measures of SES, cancer screening behaviors, and covariates. Eight employed multilevel logistic regression, whereas the remainder analyzed data with standard single level logistic regression. The majority measured one or two indicators of area and individual SES; common indicators at both levels were poverty, income, and education. There was no consistent pattern in the association between area SES and cancer screening. DISCUSSION: The gaps and conceptual and methodologic heterogeneity in the literature to date limit definitive conclusions about an underlying association between area SES and cancer screening. We identify five areas of research deserving greater attention in the literature. PMID- 19815636 TI - Predictors of marriage and divorce in adult survivors of childhood cancers: a report from the Childhood Cancer Survivor Study. AB - BACKGROUND/OBJECTIVE: Adult survivors of childhood cancer can have altered social functioning. We sought to identify factors that predict marriage and divorce outcomes in this growing population. METHODS: This was a retrospective cohort study of 8,928 > or = 5-year adult survivors of childhood malignancy and 2,879 random sibling controls participating in the Childhood Cancer Survivor Study. Marital status, current health, psychological status, and neurocognitive functioning were determined from surveys and validated instruments. RESULTS: Survivors were more likely to be never-married than siblings [relative risk (RR), 1.21; 95% confidence interval (95% CI), 1.15-1.26] and the U.S. population (RR, 1.25; 95% CI, 1.21-1.29), after adjusting for age, gender, and race. Patients with central nervous system tumors were at greatest risk of not marrying (RR, 1.50; 95% CI, 1.41-1.59). Married survivors divorced at frequencies similar to controls. In multivariable regression analysis, nonmarriage was most associated with cranial radiation (RR, 1.15; 95% CI, 1.02-1.31 for > 2,400 centigray). In analysis of neurobehavioral functioning, nonmarriage was associated with worse task efficiency (RR, 1.27; 95% CI, 1.20-1.35), but not with emotional distress, or problems with emotional regulation, memory, or organization. Physical conditions predictive of nonmarriage included short stature (RR, 1.27; 95% CI, 1.20-1.34) and poor physical function (RR, 1.08; 95% CI, 1.00-1.18). Structural equation modeling suggested that cranial radiation influenced marriage status through short stature, cognitive problems, and poor physical function. CONCLUSIONS: Childhood cancer survivors married at lower frequencies compared with peers. Patients with central nervous system tumors, cranial radiation, impaired processing efficiency, and short stature were more likely to never marry. Divorce patterns in survivors were similar to peers. PMID- 19815637 TI - Determinants and correlation of systemic and cervical concentrations of total IgA and IgG. AB - We compared systemic and cervical total IgA and IgG during the menstrual cycle among 154 women who attended clinic visits at follicular/early, periovulatory/mid, and luteal/late phases of menstrual cycle. Paired serum and cervical secretions were tested at each visit for total IgA and IgG using ELISA. Geometric mean titers for systemic IgA and IgG were 1.92 and 8.25 mg/mL, respectively. There were no differences in titers by menstrual cycle phase, neither were they correlated to cervical titers (rho = 0.17 and 0.16, respectively). The lack of correlation between systemic and cervical total IgA and IgG suggests that systemic concentrations are not reflective of cervical levels. PMID- 19815638 TI - Chromosome 3p12.3-p14.2 and 3q26.2-q26.32 are genomic markers for prognosis of advanced nasopharyngeal carcinoma. AB - PURPOSE: Nasopharyngeal carcinoma is an epithelial malignancy with a remarkable racial and geographic distribution. Previous cytogenetic studies have shown nasopharyngeal carcinoma to be characterized by gross genomic aberrations. However, identification of susceptible gene loci in advanced nasopharyngeal carcinoma has been poorly discussed. EXPERIMENTAL DESIGN: A genome-wide survey of gene copy number changes was initiated with two nasopharyngeal carcinoma cell lines by array-based comparative genomic hybridization analysis. These alterations were confirmed by a parallel analysis with the data from the gene expression microarray and were validated by quantitative PCR. Clinical association of the defined target genes was analyzed by fluorescence in situ hybridization on 48 metastatic tumors. RESULTS: A high percentage of genes were consistently altered in dosage and expression levels with gain on 3q26.2-q26.32 and losses on 3p12.3-p14.2 and 9p21.3-p23. Six candidate genes, GPR160 (3q26.2 q27), SKIL (3q26), ADAMTS9 (3p14.2-p14.3), LRIG1 (3p14), MPDZ (9p22-p24), and ADFP (9p22.1) were validated by quantitative PCR. Fluorescence in situ hybridization studies revealed amplification of GPR160 (in 25% of cases) and SKIL (33%); and deletion of ADAMTS9 (30%), LRIG1 (35%), MPDZ (15%), and ADFP (15%). Clinical association analyses indicated a poor survival rate with genetic alterations at the defined 3p deletion (P = 0.0012) and the 3q amplification regions (P = 0.0114). CONCLUSION: The combined microarray technologies suggested novel candidate oncogenes, amplification of GPR160 and SKIL at 3q26.2-q26.32, and deletion of tumor suppressor genes ADAMTS9 and LRIG1 at 3p12.3-p14.2. Altered expression of these genes may be responsible for malignant progression and could be used as potential markers for nasopharyngeal carcinoma. PMID- 19815640 TI - IGF1 and IGFBP3 polymorphisms and plasma levels in women. PMID- 19815642 TI - Keeping our eye on the ball: the American Society of Preventive Oncology in 2009. PMID- 19815643 TI - Potential effects of active parental consent: Enrolling teen smokers into a school-based cessation program. AB - INTRODUCTION: Research on effective teen smoking cessation interventions is critical to reducing the tobacco-related disease burden and risk of lifetime negative health outcomes for youth. However, informed consent procedures requiring active parental consent may restrict or influence teen participation in critical teen cessation programs. METHODS: Not On Tobacco (N-O-T) is a teen smoking cessation intervention that has been implemented under both active parental consent and passive parental consent conditions. The present study determined if there are differences in characteristics of youth enrolled under each condition. Data were available for active consent (n = 968) and passive consent (n = 4,924) participants aged 14-18 who completed the N-O-T program between 1998 and 2006 across several states. RESULTS: Participants enrolled under active consent conditions were more likely to be older, White/non-Hispanic, live in father-only or grandparent-headed household, start smoking at an earlier age, smoke more on weekdays, have previous unsuccessful quit attempts, and have siblings and friends who smoke. Additional differences were found between active and passive consent conditions in motivation to quit smoking, confidence in quitting, and stage of change. DISCUSSION: Results highlight important differences between youth who enroll in a smoking cessation program under active and passive consent conditions, often a distinguishing feature of research and non-research implementation. PMID- 19815645 TI - Gene association analysis: a survey of frequent pattern mining from gene expression data. AB - Establishing an association between variables is always of interest in genomic studies. Generation of DNA microarray gene expression data introduces a variety of data analysis issues not encountered in traditional molecular biology or medicine. Frequent pattern mining (FPM) has been applied successfully in business and scientific data for discovering interesting association patterns, and is becoming a promising strategy in microarray gene expression analysis. We review the most relevant FPM strategies, as well as surrounding main issues when devising efficient and practical methods for gene association analysis (GAA). We observed that, so far, scalability achieved by efficient methods does not imply biological soundness of the discovered association patterns, and vice versa. Ideally, GAA should employ a balanced mining model taking into account best practices employed by methods reviewed in this survey. Integrative approaches, in which biological knowledge plays an important role within the mining process, are becoming more reliable. PMID- 19815644 TI - Control of ovulation in mice by progesterone receptor-regulated gene networks. AB - The mid-cycle surge of luteinizing hormone (LH) induces ovulation, a process during which a fertilizable oocyte is released from a mature ovarian follicle. Although ovulation is a physiologically well-characterized event, the underlying molecular pathways remain poorly understood. Progesterone receptor (PGR), which mediates the biological effects of the steroid hormone progesterone, has emerged as a key regulator of ovulation in mice. The development of a progesterone receptor-null (Pgr-null) mouse model confirmed a critical role of this hormone in ovulation because in these mutant mice, mature pre-ovulatory follicles fail to release the oocytes. This animal model has thus presented a unique opportunity to study the molecular pathways underlying ovulation. Gene-expression profiling experiments by several groups, using the ovaries of Pgr-null mice, revealed novel gene networks, which act downstream of PGR to control ovulation. These genes encode diverse molecules such as proteases, transcription factors, cell-adhesion molecules, modulators of vascular activities and regulators of inflammation. Functional analyses using gene-knockout mouse models have confirmed that some of these factors play critical roles during ovulation. The knowledge gained from these studies has helped us to understand better the molecular mechanisms that facilitate the release of oocytes from pre-ovulatory follicles. Further analysis of the role of molecular regulators of ovulation will help identify useful molecular targets that would allow the development of improved contraceptives and new therapeutics for anovulatory infertility. PMID- 19815646 TI - Social and community participation of children and youth with cerebral palsy is associated with age and gross motor function classification. AB - BACKGROUND: Through social and community participation, children and youth with cerebral palsy (CP) form friendships, gain knowledge, learn skills, express creativity, and determine meaning and purpose in life. OBJECTIVE: The purposes of this study were: (1) to determine whether social and community participation of children and youth with CP differ based on age, sex, and gross motor function, and (2) to identify the types of activities in which social and community participation are highest. DESIGN AND METHODS: A prospective cross-sectional analytic design was used. The participants were a sample of convenience of 291 children (6-12 years of age) and 209 youth (13-21 years of age) with CP (55.4% males, 44.6% females) receiving services from 7 children's hospitals. Participants completed the Children's Assessment of Participation and Enjoyment (CAPE) by structured interview. Gross Motor Function Classification System (GMFCS) level was determined by the researchers. RESULTS: Youth did a higher percentage of activities with friends and others and outside the home than children. Children and youth in level I did a higher percentage of activities with friends and others compared with children and youth in levels II and III and in levels IV and V. Children and youth in level I and in levels IV and V did a higher percentage of activities outside the home than children and youth in levels II and III. Differences were not found between females and males. The percentage of activities done with friends and others and outside the home was highest for physical and skill-based activities. LIMITATIONS: Findings cannot be attributed only to GMFCS level. CONCLUSIONS: The ability to walk without restrictions is desirable for social and community participation. For children and youth with CP who have limitations in mobility, physical therapists have roles as consultants for accessibility, activity accommodations, and assistive technology and as advocates for inclusive environments. PMID- 19815647 TI - Infants born preterm exhibit different patterns of center-of-pressure movement than infants born at full term. AB - BACKGROUND: Infants born preterm are at risk for developmental impairments related to postural control. OBJECTIVE: The purpose of this study was to determine whether infants born preterm and infants born at full term differed in postural control at 1 to 3 weeks after term age. DESIGN: This study included 17 infants born preterm (mean gestational age=31.9 weeks, range=25.0-34.6) and 15 infants born at full term (mean gestational age=38.9 weeks, range=37.3-40.6). All infants were without diagnosed neurological or genetic conditions. MEASUREMENT: Center-of-pressure (COP) data were recorded at 5 Hz while each infant was positioned supine on a pressure-sensitive mat in an alert behavioral state. Root mean square (RMS) displacement and approximate entropy (ApEn) were used to describe the COP movement variability in the time series. Differences between groups were identified using independent t tests. RESULTS: The COP time series were found to be deterministic, suggesting order in the time series. Infants born preterm exhibited significantly larger RMS values in the caudal-cephalic direction than infants born at full term (1.11 and 0.83 cm, respectively; t=-2.6, df=30, P=.01). However, infants born at full term had significantly larger ApEn values in the caudal-cephalic direction (1.19 and 1.11, respectively; t=2.4, df=30, P=.02). The 2 groups did not differ in RMS or ApEn values in the medial lateral direction or the resultant. CONCLUSIONS: Infants born at full term exhibited COP displacements in the caudal-cephalic direction that were smaller in amplitude, but may be considered more complex or less predictable, than those of infants born preterm. One explanation is that infants born preterm exhibited more stereotypic patterns of movement, resulting in large, but repetitive, COP excursions. A combination of linear and nonlinear measures may provide insight into the control of posture of young infants. PMID- 19815648 TI - Antinociceptive Activity of Trichilia catigua Hydroalcoholic Extract: New Evidence on Its Dopaminergic Effects. AB - Trichilia catigua is a native plant of Brazil; its barks are used by some local pharmaceutical companies to prepare tonic drinks, such as Catuama. The present study was addressed to evaluate the effects of T. catigua hydroalcoholic extract in mouse nociception behavioral models, and to evaluate the possible mechanisms involved in its actions. Male Swiss mice were submitted to hot-plate, writhing and von Frey tests, after oral treatment with T. catigua extract (200 mg kg(-1), p.o.). The extract displayed antinociceptive effect in all three models. For characterization of the mechanisms involved in the antinociceptive action of the extract, the following pharmacological treatments were done: naloxone (2.5 mg kg( 1), s.c.), SR141716A (10 mg kg(-1), i.p.), SCH23390 (15 MUg kg(-1), i.p.), sulpiride (50 mg kg(-1), i.p.), prazosin (1 mg kg(-1), i.p.), bicuculline (1 mg kg(-1), i.p.) or dl-p-chlorophenylalanine methyl ester (PCPA, 100 mg kg(-1), i.p.). In these experiments, the action of T. catigua extract was evaluated in the hot-plate test. The treatment with SCH23390 completely prevented the antinociceptive effect, while naloxone partially prevented it. The possible involvement of the dopaminergic system in the actions of T. catigua extract was substantiated by data showing the potentiation of apomorphine-induced hypothermia and by the prevention of haloperidol-induced catalepsy. In conclusion, the antinociceptive effects of T. catigua extract seem to be mainly associated with the activation of dopaminergic system and, to a lesser extent, through interaction with opioid pathway. PMID- 19815649 TI - An open-label expanded access study of lapatinib and capecitabine in patients with HER2-overexpressing locally advanced or metastatic breast cancer. AB - BACKGROUND: The Lapatinib Expanded Access Program (LEAP) was designed to provide access to lapatinib plus capecitabine for HER2-positive metastatic breast cancer patients who previously received an anthracycline, a taxane, and a trastuzumab and had no other treatment options. PATIENTS AND METHODS: LEAP opened globally and enrollment continued until lapatinib received regulatory approval in each participating country. Patients were assessed for progression-free survival (PFS) and overall survival (OS) and monitored for serious adverse events (SAEs). RESULTS: As of 30 September 2008, 4283 patients from 45 countries enrolled in LEAP. The median treatment duration was 24.7 weeks. The most common drug-related SAEs were diarrhea (9.7%), vomiting (4.3%), and nausea (2.4%) and were mainly grade 3 or higher. The incidences of special interest SAEs were decreased left ventricle ejection fraction (0.5%), interstitial lung disease/pneumonitis (0.2%), and serious hepatobiliary events (0.4%). This safety profile is consistent with the overall lapatinib program. The median PFS and OS were 21.1 [95% confidence interval (CI) = 20.1-22.3] and 39.6 (95% CI = 37.7-40.7) weeks, respectively (n = 4006). Subgroup analysis showed longer PFS and OS in patients who had not received prior capecitabine. CONCLUSIONS: These results demonstrate the safety and efficacy of lapatinib in a broader patient population compared with a clinical trial. PMID- 19815650 TI - Global drug development in cancer: a cross-sectional study of clinical trial registries. AB - BACKGROUND: Drug development in cancer is costly and may be directed toward 'profitable' cancers of the more developed regions (MDR) as compared with those of the less developed regions (LDR) of the world. Here, we describe drug development in relation to cancer type and geographic location. MATERIALS AND METHODS: We reviewed phase II and III clinical trials evaluating new cancer drugs, which were registered from January to June 2008. Correlations were sought between the number of clinical trials and incidence, mortality and prevalence of the cancers studied (obtained from GLOBOCAN 2002) and stratified by region of the world. RESULTS: We identified 399 newly registered trials. Most trials (N = 229, 57%) were sponsored by industry. The most common types of cancer studied were breast 73 (18%), lung 57 (14%), prostate 44 (11%) and colorectal 28 (7%). In MDR, incidence, mortality and prevalence correlated significantly (Pearson r = 0.80, 0.73 and 0.63; P < or = 0.01) with the number of all registered clinical trials, whereas in LDR, only prevalence showed significant association (Pearson r = 0.55; P = 0.03) with the number of trials for a given type of cancer. CONCLUSION: Lethal cancers that are common in the LDR (e.g. stomach, liver and esophageal cancers) deserve greater emphasis for drug development. PMID- 19815651 TI - Randomized phase II trial of weekly paclitaxel combined with carboplatin versus standard paclitaxel combined with carboplatin for elderly patients with advanced non-small-cell lung cancer. AB - BACKGROUND: The optimal platinum doublet regimen in elderly patients with non small-cell lung cancer (NSCLC) is still uncertain. We conducted a randomized phase II study to compare the efficacy and safety of weekly paclitaxel combined with carboplatin with those of the standard schedule. PATIENTS AND METHODS: Elderly patients (age > or =70 years) with advanced NSCLC were randomly assigned to either the weekly arm {70 mg/m(2) paclitaxel on days 1, 8, and 15 and carboplatin [area under the curve (AUC) = 6] on day 1} or the standard arm [200 mg/m(2) paclitaxel and carboplatin (AUC = 6) on day 1]. The primary end point was the overall response rate (ORR). RESULTS: Eighty-two patients were enrolled. The ORR and median progression-free survival were 55% and 6.0 months for the weekly arm and 53% and 5.6 months for the standard arm. Grade 3/4 neutropenia and peripheral neuropathy were observed in 41% and 0% of the patients in the weekly arm and in 88% and 25% in the standard arm, respectively. CONCLUSIONS: This is the first randomized study that compares the platinum doublet designed specifically for the elderly. Regarding the safety, the weekly regimen was less toxic than the standard regimen and seems to be preferable for elderly patients with advanced NSCLC. PMID- 19815654 TI - Case study: multisystemic therapy for adolescents who engage in HIV transmission risk behaviors. AB - OBJECTIVE: To present a case study using multisystemic therapy (MST), an intensive family focused psychotherapy. For the clinical trial from which this case was drawn, MST was adapted to address multiple human immunodeficiency virus (HIV) transmission risk behaviors in HIV-infected youth. Targeted behaviors included medication nonadherence, risky sexual behaviors, and substance use. METHOD: One young woman's transmission risk behaviors are described, followed by a description of the MST procedures used to identify and treat the primary drivers of these risk behaviors. Outcome measures were self-report, urine screens, and blood draws. RESULTS: At discharge, the young woman showed significant improvements in medication adherence and related health status (e.g., reduced HIV viral load), healthier sexual behaviors, and reduced substance use. Importantly, neither her boyfriend nor her newborn tested positive for HIV. CONCLUSIONS: Findings from this case study suggest that MST has the potential to reduce transmission risk behaviors among teens with HIV. PMID- 19815655 TI - Communicating contentious health policy: lessons from Ireland's workplace smoking ban. AB - The Irish workplace smoking ban has been described as possibly a tipping point for public health worldwide. This article presents the first analysis of the newspaper coverage of the ban over the duration of the policy formation process. It adds to previous studies by analyzing how health communication strategists engaged, over time, with a newsworthy topic, viewed as being culturally controversial. It analyzes a sample of media content (n = 1,154) and firsthand accounts from pro-ban campaigners and journalists (n = 10). The analysis shows that the ban was covered not primarily as a health issue: Economic, political, social, democratic, and technical aspects also received significant attention. It shows how coverage followed controversy and examines how pro-ban campaigners countered effectively the anti-ban communication efforts of influential social actors in the economic and political spheres. The analysis demonstrates that medical-political sources successfully defined the ban's issues as centrally concerned with public health. PMID- 19815652 TI - Preoperative weekly cisplatin, epirubicin, and paclitaxel (PET) improves prognosis in locally advanced breast cancer patients: an update of the Southern Italy Cooperative Oncology Group (SICOG) randomised trial 9908. AB - BACKGROUND: The present article reports the updated survival outcome of the 200 patients enrolled in the Southern Italy Cooperative Oncology Group 9908 trial, which compared 12 weekly cycles of cisplatin-epirubicin-paclitaxel (PET) with 4 triweekly (once every 3 weeks) cycles of epirubicin-paclitaxel (ET) in patients with locally advanced breast cancer (LABC). METHODS: The effects of treatment, pathologically documented response (pathological response), pre- and post treatment biomarkers on relapse-free survival (RFS), distant metastasis-free survival (DMFS), and overall survival (OS) are analysed. RESULTS: At a median follow-up of 74 (range 48-105 months) months, the 5-year RFS, DMFS, and OS were 64 % versus 53% (P = 0.11), 73% versus 55% (P = 0.04), and 82% versus 69% (P = 0.07) in PET and ET, respectively. At multivariate analysis, after adjusting treatment effect for pretreatment biomarkers, PET independently predicted better DMFS (P = 0.018) and OS (P = 0.03), whereas the impact on RFS was of borderline significance (0.057). PET treatment was significantly better than ET treatment only in high-grade or highly proliferating tumours. The better outcome in PET arm was the results of both the higher rate of patients with optimal pathological response and the lower rate of patients with biologically aggressive residual tumour. CONCLUSIONS: The PET weekly regimen significantly improves both DMFS and OS in LABC patients, compared with the triweekly ET combination. The therapeutic advantage is limited to patients with highly aggressive tumours. PMID- 19815656 TI - Establishing cause for developing and implementing a broad-based marketing effort for the health education field. AB - What exactly is health education? Professionals with advanced degrees in health education have most likely encountered questions such as these either during introductory coursework or from those inquiring about the field. These queries can prove quite perplexing when asked by individuals who are unaware of the health education profession. Because the act of marketing health education is crucial to the sustainability of the field, the purpose of this article is to (a) explore the issue of describing and promoting health education, (b) establish ideas that can facilitate the provision of coordinated marketing efforts, and (c) offer marketing management and implementation principles that can assist in marketing both health education and health educators. Based on this discussion, the authors suggest building mainstream consensus in regards to marketing message development and implementation to better position health education. PMID- 19815657 TI - Diagnostic agreement in the assessment of orthodontic treatment need using the Dental Aesthetic Index and the Index of Orthodontic Treatment Need. AB - The aim of this study was to estimate the diagnostic agreement between assessments of orthodontic treatment need of a child population using the Dental Aesthetic Index (DAI) and the Index of Orthodontic Treatment Need (IOTN). A cross sectional study of a representative random sample of children aged 12 (n = 475) and 15-16 (n = 398) years was carried out in the Valencia region of Spain. A Student's t-test was used to compare the DAI means by gender and age and a chi square test to compare the proportions of the population in need of orthodontic treatment. To calculate the agreement between the two indices, intra-class correlation coefficient and Kappa statistics were employed. Of the 12-year-olds, 23.5 percent (n = 121) and of the 15- to 16-year-olds, 26.6 percent (n = 108) were receiving or had previously received orthodontic treatment. The observed agreement between the two indices on the need for treatment among the 12-year olds (n = 363) was 83.4 percent and Kappa for diagnostic agreement was 0.52 [95 percent confidence interval (CI): 0.42-0.63]. For the 15- to 16-year-olds (n = 292), the figures were 82.5 percent and 0.38 (95 percent CI: 0.24-0.52), respectively. For the total sample (n = 655), the observed agreement was 83 percent and the diagnostic agreement was 0.47 (95 percent CI: 0.39-0.55). For this population, there was only moderate agreement between the two indices. This means that, when one of these indices is used to measure or prioritize orthodontic treatment in a determined population, the individuals selected with an obvious treatment need are going to be different in 17 percent of the cases depending on which index is used, DAI or IOTN. This difference has to be taken in consideration when measuring, recording, or quantifying orthodontic treatment need. PMID- 19815658 TI - Direct activation of protein kinases by ubiquitin. AB - The inhibitor of kB kinase (IKK) complex, a critical positive regulator of nuclear factor kappaB (NF-kappaB) signaling and immune function, can be activated in vitro by polyubiquitin chains. The modification of signaling intermediates by polyubiquitin and their association with ubiquitin-binding domain-containing proteins within multimolecular signaling complexes therefore represent a potential mechanism for activation of IKK and NF-kappaB in vivo. PMID- 19815659 TI - Uncertainty assessment of input parameters for economic evaluation: Gauss's error propagation, an alternative to established methods. AB - In decision modeling for health economic evaluation, bootstrapping and the Cholesky decomposition method are frequently used to assess parameter uncertainty and to support probabilistic sensitivity analysis. An alternative, Gauss's error propagation law, is rarely known but may be useful in some settings. Bootstrapping, the Cholesky decomposition method, and the error propagation law were compared regarding standard deviation estimates of a hypothetic parameter, which was derived from a regression model fitted to simulated data. Furthermore, to demonstrate its value, the error propagation law was applied to German administrative claims data. All 3 methods yielded almost identical estimates of the standard deviation of the target parameter. The error propagation law was much faster than the other 2 alternatives. Furthermore, it succeeded the claims data example, a case in which the established methods failed. In conclusion, the error propagation law is a useful extension of parameter uncertainty assessment. PMID- 19815660 TI - Serum levels of large tenascin-C variants, matrix metalloproteinase-9, and tissue inhibitors of matrix metalloproteinases in concentric versus eccentric left ventricular hypertrophy. AB - AIMS: Chronic hypertension may cause left ventricular hypertrophy (LVH). The role of matrix metalloproteinases (MMPs), tissue inhibitors of matrix metalloproteinases (TIMPs), and tenascin-C (Tn-C) splice variants in concentric vs. eccentric left ventricular remodelling has not been investigated. METHODS AND RESULTS: Serum levels of B or C domain containing Tn-C, MMP-9, TIMP-1, -2, and -4 were determined in concentric (left ventricular posterior wall thickness >13 mm and intraventricular septum >13 mm, n = 61) and eccentric (end-diastolic left ventricular diameter >55 mm or end-systolic left ventricular diameter >40 mm, n = 34) LVH by enzyme-linked immunoassays. Levels of B domain containing Tn-C were higher in patients with LVH than in normal volunteers (P = 0.020) and higher in eccentric LVH (EH) compared with concentric LVH (CH) (P = 0.003). A cut-off value of 900 ng/mL might discriminate between these different forms of LVH. Matrix metalloproteinase-9 was higher in patients with LVH than in normal volunteers (P = 0.042), and levels were decreased in EH compared with CH (P = 0.028). Patients with LVH had higher levels of TIMP-1 (P = 0.059), TIMP-2 (P = 0.043), and TIMP-4 (P = 0.163) than normal volunteers, but there were no differences between the LVH groups. CONCLUSION: Our data suggest that myocardial remodelling in LVH is associated with changes in serum levels of MMP-9, TIMP-1, -2, -4, and Tn-C splice variants. In addition, B domain containing Tn-C discriminated EH from CH and might be suggested as a potential diagnostic marker. PMID- 19815661 TI - The safety and tolerability of darbepoetin alfa in patients with anaemia and symptomatic heart failure. AB - AIMS: To assess the safety and tolerability of darbepoetin alfa (DA) in the treatment of anaemia in heart failure (HF). METHODS AND RESULTS: In this pooled analysis of three randomized, double-blind, placebo-controlled studies of anaemic [haemoglobin (Hb) < or =12.0 g/dL or < or =12.5 g/dL] symptomatic HF subjects, DA was administered subcutaneously once every 2 weeks and titrated to achieve and maintain a target Hb of 14.0 +/- 1.0 g/dL. In total, 516 subjects were randomized; 231 (44.8%) to placebo, 285 (55.2%) to DA. Darbepoetin alfa was well tolerated, with an adverse event (AE) profile similar to placebo. Most subjects (placebo, 85%; DA, 87%) experienced at least one AE. There was a lower incidence of serious AEs in the DA group (placebo, 43%; DA, 37%) with the most frequent being worsening HF (placebo, 19%; DA, 11%). Treatment-related AEs were reported for 9% and 12% in placebo and DA subjects, respectively. Fewer deaths were reported in DA group (6%) vs. placebo (8%). CONCLUSION: Darbepoetin alfa was well tolerated with an AE profile similar to placebo in HF subjects treated to a target Hb of 14.0 +/- 1.0 g/dL. Contrary to recent data in other patient populations, there was no evidence of increased risk of mortality or cardiovascular events. PMID- 19815662 TI - Comparison of nasopharyngeal nylon flocked swabs with universal transport medium and rayon-bud swabs with a sponge reservoir of viral transport medium in the diagnosis of paediatric influenza. AB - This study compared a kit containing a nasopharyngeal nylon flocked swab and a tube with a liquid universal transport medium (UTM) with a kit containing a plastic-shafted rayon-budded swab with a sponge reservoir of viral transport medium for the molecular detection of influenza viruses in children. Respiratory samples were collected from 314 children aged <5 years with influenza-like illness (186 males; mean age 2.32+/-2.27 years) using both swabs in a randomized sequence for each patient. The flocked swabs permitted the detection of 28 influenza A (8.9 %) and 45 influenza B (14.3 %) cases, and the rayon-bud swabs 26 influenza A (8.3 %) and 43 influenza B (13.7 %) cases, with detection rates of 23.2 and 22.0 %, respectively, and similar cycle threshold values. Paediatricians and laboratory staff were significantly more satisfied with both the simplicity (P <0.0001) and rapidity (P <0.0001) of the nasopharyngeal flocked swabs with UTM. These findings show that the flocked swabs with UTM and the rayon-bud swabs with a sponge transport medium are similarly efficient in preserving influenza virus nucleic acid, but that the kit containing a flocked swab with a UTM allows easier and more rapid collection and processing of specimens. PMID- 19815663 TI - Visualization of adherent micro-organisms using different techniques. AB - The visualization and quantification of adherent bacteria is still one of the most relevant topics in microbiology. Besides electron microscopic techniques such as transmission electron microscopy, scanning electron microscopy and environmental scanning electron microscopy, modern fluorescence microscopic approaches based on fluorogenic dyes offer detailed insight into bacterial biofilms. The aim of the present review was to provide an overview of the advantages and disadvantages of different methods for visualization of adherent bacteria with a special focus on the experiences gained in dental research. PMID- 19815664 TI - Diversity in the cag pathogenicity island of Helicobacter pylori isolates in populations from North and South India. AB - The cag pathogenicity island (cagPAI) has been reported to be the major virulence determinant in Helicobacter pylori-related diseases. In the present study, the diversity of the cagA gene and the integrity of the cagPAI in 158 H. pylori strains from Varanasi (North India) and Hyderabad (South India) were studied by amplifying the cagA gene (approximately 3.5 kb), followed by PCR-RFLP analysis. The results revealed significant differences in the cagA gene and the integrity of the cagPAI between North and South Indian isolates. Of 158 isolates, 40 (34.8 %) from Varanasi and 20 (46.5 %) from Hyderabad were found to carry an intact cagPAI. A partially deleted cagPAI was present in 75 (65.2 %) isolates from Varanasi and 23 (53.5 %) from Hyderabad. None of the isolates showed complete deletion of the cagPAI. Differences in the cagA 5' and 3' regions were also noted, and 11 isolates (8 from Varanasi and 3 from Hyderabad) that were cagA negative with primers for the 5' region turned out to be cagA positive with primers for the 3' variable region. It is tentatively concluded that the 3' variable region may be a better marker for cagA typing. The results also showed that the majority of the isolates harboured the Western-type EPIYA motif. PCR RFLP analysis of the cagA gene showed 29 distinguishable digestion patterns, and cluster analysis of RFLP types from a random selection of 32 isolates placed all of the isolates into 5 groups. These results demonstrate that significant differences in the cagPAI occur among isolates from North and South India, and that RFLP of cagA could be employed for elucidating genetic variations among various isolates of H. pylori. PMID- 19815665 TI - Vibrio cholerae O1 Ogawa detoxified lipopolysaccharide structures as inducers of cytokines and oxidative species in macrophages. AB - Multidrug resistance in several strains of Vibrio cholerae has encouraged anti cholera vaccine developmental attempts using various subcellular moieties. In order to examine the immunological efficacy of detoxified LPS (dLPS)-derived saccharide immunogens, ex vivo activation of mouse peritoneal macrophages (MPhis) was investigated. The immunomodulatory effect was evaluated via induction of the pro-inflammatory cytokines tumour necrosis factor-alpha, interleukin (IL)-1 alpha and IL-6 and acceleration of nitric oxide (NO) and reactive oxygen species (ROS). Immunologically active structures triggered mouse peritoneal MPhis to secrete cytokines and release NO/ROS, even at concentrations as low as 12.5 microg ml( 1). It was found that the O-specific polysaccharide moiety was more immunologically efficient than the glycolipid one, probably due to the position of 3-deoxy-D-manno-octulosonic acid. The results revealed effective structure immunomodulating relationships of dLPS-derived moieties that are desirable in subcellular anti-cholera vaccine design. PMID- 19815666 TI - Mycoplasma salivarium detected in a microbial community with Candida glabrata in the biofilm of an occluded biliary stent. AB - Mycoplasma salivarium, preferentially an inhabitant of the human oral cavity, has rarely been found in other locations associated with disease. We describe here, for what is believed to be the first time, the detection of M. salivarium, together with Candida glabrata, in an occluded biliary stent of an icteric, cholestatic patient. PMID- 19815667 TI - Profiling of mismatch discrimination in RNAi enabled rational design of allele specific siRNAs. AB - Silencing specificity is a critical issue in the therapeutic applications of siRNA, particularly in the treatment of single nucleotide polymorphism (SNP) diseases where discrimination against single nucleotide variation is demanded. However, no generally applicable guidelines are available for the design of such allele-specific siRNAs. In this paper, the issue was approached by using a reporter-based assay. With a panel of 20 siRNAs and 240 variously mismatched target reporters, we first demonstrated that the mismatches were discriminated in a position-dependent order, which was however independent of their sequence contexts using position 4th, 12th and 17th as examples. A general model was further built for mismatch discrimination at all positions using 230 additional reporter constructs specifically designed to contain mismatches distributed evenly along the target regions of different siRNAs. This model was successfully employed to design allele-specific siRNAs targeting disease-causing mutations of PIK3CA gene at two SNP sites. Furthermore, conformational distortion of siRNA target duplex was observed to correlate with the compromise of gene silencing. In summary, these findings could dramatically simplify the design of allele-specific siRNAs and might also provide guide to increase the specificity of therapeutic siRNAs. PMID- 19815668 TI - A simple method for directional transcriptome sequencing using Illumina technology. AB - High-throughput sequencing of cDNA has been used to study eukaryotic transcription on a genome-wide scale to single base pair resolution. In order to compensate for the high ribonuclease activity in bacterial cells, we have devised an equivalent technique optimized for studying complete prokaryotic transcriptomes that minimizes the manipulation of the RNA sample. This new approach uses Illumina technology to sequence single-stranded (ss) cDNA, generating information on both the direction and level of transcription throughout the genome. The protocol, and associated data analysis programs, are freely available from http://www.sanger.ac.uk/Projects/Pathogens/Transcriptome/. We have successfully applied this method to the bacterial pathogens Salmonella bongori and Streptococcus pneumoniae and the yeast Schizosaccharomyces pombe. This method enables experimental validation of genetic features predicted in silico and allows the easy identification of novel transcripts throughout the genome. We also show that there is a high correlation between the level of gene expression calculated from ss-cDNA and double-stranded-cDNA sequencing, indicting that ss-cDNA sequencing is both robust and appropriate for use in quantitative studies of transcription. Hence, this simple method should prove a useful tool in aiding genome annotation and gene expression studies in both prokaryotes and eukaryotes. PMID- 19815669 TI - The Thomsonian movement, the regular profession, and the state in antebellum Connecticut: a case study of the repeal of early medical licensing laws. AB - The Thomsonian movement, founded by Samuel Thomson, was the first major challenge to the therapies and the social and economic standing of the orthodox medical profession in the United States. In the late-eighteenth or early-nineteenth century, many states chartered a state medical society with power to administer a licensing law that placed at least a nominal penalty on practicing without a license. However, in the 1830s and 1840s, under pressure by proponents of the Thomsonian system, almost all legislatures reversed themselves and removed all restrictions on medical practice. This paper reexamines the rise and fall of medical licensing using Connecticut as a case study. Antebellum legislative controversies over licensing have never been described in detail at the state level--where the drama took place--integrating the perspectives of both the medical regulars and Thomsonian botanical physicians, and state politics. Connecticut is a particularly useful case study because, except for New York, its seven-year battle from 1836 to 1842 over the medical society's charter was the most protracted in the country. How was the campaign structured? To what extent did the licensing restrictions matter? What role did the state-level Democratic party play? Thomsonianism in Connecticut, I suggest, was more professionalized and conservative than historians have often portrayed this movement. This account shows that the state's Thomsonian physicians were not anti-professional or opposed to education, but rather used the politics of the antebellum era to challenge the medical law and legitimize themselves as an alternative form of practice. PMID- 19815672 TI - Balanced and responsible journalism. PMID- 19815671 TI - CT60 and +49 polymorphisms of CTLA 4 are associated with ANCA-positive small vessel vasculitis. AB - OBJECTIVE: To investigate whether single nucleotide polymorphisms (SNPs) within cytotoxic T-lymphocyte antigen-4 (CTLA-4) are associated with ANCA-associated small vessel vasculitis (SVV). METHODS: The CTLA-4 CT60 (exon 4), +49 (exon 1) and -318 (promoter region) genotypes were determined by PCR and restriction fragment length polymorphism (RFLP) in 222 white Caucasians of UK origin with SVV and 670 ethnically matched controls. RESULTS: The CTLA-4 exon 1 (+49) and 4 (CT60) polymorphisms are associated with SVV (+49: chi(2) = 10.965, P = 0.004; CT60: chi(2) = 12.017, P = 0.002). Both disease-susceptible and disease protective haplotypes have been identified in this cohort, and their frequencies are similar in the subtypes of WG and microscopic polyangiitis. CONCLUSION: This study provides further evidence that CTLA-4, a susceptibility locus for a number of common autoimmune diseases, may also be involved in the development of ANCA associated SVV. PMID- 19815670 TI - Investigating the role of the interleukin-23/-17A axis in rheumatoid arthritis. AB - OBJECTIVE: IL-23 is a pro-inflammatory cytokine proposed to be central to the development of autoimmune disease. We investigated whether IL-23, together with the downstream mediator IL-17A, was present and functional in RA in humans. METHODS: RA synovial cells were cultured in the presence or absence of antibodies directed against IL-23p19 or -23R and -17. IL-23, -12, -17, and their receptors, and IL-6, -1beta and TNF-alpha were measured by ELISA and/or PCR. RESULTS: Small amounts of cell-associated IL-23 (median 110 pg/ml) were detected in RA synovial cultures, and found to be functional as IL-23R blockade resulting in a significant inhibition of TNF-alpha (57%), IL-1beta (51%) and IL-6 (30%). However, there was a considerable variability between individual patient samples, and anti-IL-23p19 was found to be considerably less effective. IL-17A protein was detected in approximately 40% of the supernatants and IL-17A blockade, in IL-17A producing cultures, resulted in a small but significant inhibition of TNF-alpha (38%), IL-1beta (23%) and IL-6 (22%). Addition of recombinant IL-23 to cultures had a variable effect on the spontaneous production of endogenous IL-17A with enhancement observed in some but not all cultures, suggesting that either the low levels of endogenous IL-23 are sufficient to support cytokine production and/or that the relevant Th17 cells were not present. CONCLUSIONS: These results suggest that although IL-23 may have pathogenic activity in a proportion of patients with late-stage RA, it is not abundantly produced in this inflammatory tissue, nor does it have a dominant role in all patient tissues analysed. PMID- 19815673 TI - Epidemiological usefulness of population-based electronic clinical records in primary care: estimation of the prevalence of chronic diseases. AB - BACKGROUND: Chronic health problems are the main cause of disease, disability and death in developed countries, and their prevalence is increasing. OBJECTIVE: To estimate the prevalence of selected chronic illnesses based on electronic clinical records in primary care (ECRPC) and to assess its usefulness for epidemiological research, by comparing ECRPC data against those reported by a contemporary health survey. METHODS: DESIGN: Descriptive cross-sectional study. SETTING: All primary care medical consultations in the Madrid Regional Public Health System (PHS). SUBJECTS: A total of 23 535 182 ECRPC-registered episodes of illness, generated by PHS patients over 15 years of age seeking medical care during 2005-06. MAIN OUTCOME MEASURES: Prevalences of chronic diseases estimated on the basis of medically examined cases registered in ECRPC and morbidity as reported by a contemporary health survey covering the same geographic area. RESULTS: A total of 52.5% of the adult population had some chronic health problem. The highest overall prevalences were hypertension (14.8%), mental disorders (12.0%) and allergy (11.6%). Prevalences were generally highest among women, elderly and the native population. Depending on the specific disease, ECRPC-based prevalences were similar to (e.g. diabetes), higher (e.g. chronic skin problems) or lower (e.g. asthma and dyslipidaemia) than those reported by surveys, with certain age- and sex-related variations. CONCLUSIONS: Prevalences estimated from ECRPC and survey data present variations depending on the disease, age and sex. Both data sources provide complementary information about chronic disease prevalence. ECRPC have the advantage of generating an ongoing standardized register and entailing no additional effort for health professionals. PMID- 19815674 TI - Pulse oximetry in family practice: indications and clinical observations in patients with COPD. AB - PURPOSE: To establish situations in which family physicians (FPs) consider pulse oximetry a valuable addition to their clinical patient assessment; to explore pulse oximetry results (SpO(2)) when used by FPs in patients with chronic obstructive pulmonary disease (COPD); to explore associations between SpO(2) and other markers of COPD severity. METHODS: We performed three separate studies: (i) interviews plus a Delphi consensus procedure with FPs experienced in using pulse oximetry to elucidate indications for pulse oximetry; (ii) analysis of SpO(2) and clinical data in COPD patients who presented to FPs with deteriorating symptoms and (iii) analysis of SpO(2), spirometry and clinical data in patients with stable COPD. RESULTS: Interviewed FPs (n = 11) used their pulse oximeter for a range of acute (14) and non-acute (11) indications but valued it highest in acute (worsening of) dyspnoea, in suspected respiratory insufficiency/failure and in patients with COPD. In 88 patients with deteriorating COPD, 22% showed SpO(2) 1) selection. We show that current models used to estimate omega are substantially biased by naturally occurring sequence compositions. We present a novel model that weights substitutions by conditional nucleotide frequencies and which escapes these artifacts. Applying it to the genomes of pathogens causing malaria, leprosy, tuberculosis, and Lyme disease gave significant discrepancies in estimates with approximately 10-30% of genes affected. Our work has substantial implications for how vaccine targets are chosen and for studying the molecular basis of adaptive evolution. PMID- 19815688 TI - Developmental and molecular physiological evidence for the role of phosphoenolpyruvate carboxylase in rapid cotton fibre elongation. AB - Cotton fibres are hair-like single-cells that elongate to several centimetres long after their initiation from the ovule epidermis at anthesis. The accumulation of malate, along with K+ and sugars, is thought to play an important role in fibre elongation through osmotic regulation and charge balance. However, there is a lack of evidence for or against such an hypothesis. Phosphoenolpyruvate carboxylase (PEPC) is a key enzyme responsible for the synthesis of malate. The potential role of PEPC in cotton fibre elongation is examined here. Developmentally, PEPC activity was higher at the rapid elongation phase than that at the slow elongation stage. Genotypically, PEPC activity correlated positively with the rate of fibre elongation and the final fibre length attained. Importantly, suppression of PEPC activity by LiCl that reduces its phosphorylation status decreased fibre length. To examine the molecular basis underlying PEPC activity, two cDNAs encoding PEPC, GhPEPC1 and 2, were cloned, which represents the major PEPC genes expressed in cotton fibre. RT-PCR analyses revealed that GhPEPC1 and 2 were highly expressed at the rapid elongation phase but weakly at the slow-to-terminal elongation period. In situ hybridization detected mRNA of GhPEPC1 and 2 in 1 d young fibres but not in the ovule epidermis prior to fibre initiation. Collectively, the data indicate that cotton fibre elongation requires high activity of PEPC, probably through the expression of the GhPEPC1 and 2 genes. PMID- 19815690 TI - Cell volume decrease as a link between azaspiracid-induced cytotoxicity and c-Jun N-terminal kinase activation in cultured neurons. AB - Azaspiracids (AZAs) are a group of marine toxins recently described that currently includes 20 members. Not much is known about their mechanism of action, although the predominant analog in nature, AZA-1 targets several organs in vivo, including the central nervous system, and exhibits high neurotoxicity in vitro. AZA distribution is increasing globally with mussels being most widely implicated in AZA-related food poisoning events, with human poisoning by AZAs emerging as an increasing worldwide problem in recent years. We used pharmacological tools to inhibit the cytotoxic effect of the toxin in primary cultured neurons. Several targets for AZA-induced neurotoxicity were evaluated. AZA-1 elicited a concentration-dependent hyperpolarization in cerebellar granule cells of 2-3 days in vitro; however, it did not modify membrane potential in mature neurons. Furthermore, in immature cells, AZA-1 decreased the membrane depolarization evoked by exposure of the neurons to 50mM K(+). Preincubation of the neurons with 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), 4-acetamido-4' isothiocyanato-2,2'-stilbenedisulfonic acid (SITS), 5-nitro-2-(3 phenylpropylamino)benzoic acid (NPPB), amiloride, or ouabain before addition of AZA-1 decreased the AZA-1-induced neurotoxicity and the increase in phosphorylated c-Jun-N-terminal kinase (JNK) caused by the toxin, indicating that disruption in ion fluxes was involved in the neurotoxic effect of AZA-1. Furthermore, short exposures of cultured neurons to AZA-1 caused a significant decrease in neuronal volume that was reverted by preincubation of the neurons with DIDS or amiloride before addition of the toxin. The results presented here indicate that the JNK activation induced by AZA-1 is secondary to the decrease in cellular volume elicited by the toxin. PMID- 19815691 TI - Expression of tyrosine hydroxylase increases the resistance of human neuroblastoma cells to oxidative insults. AB - In this study, we demonstrate that human neuroblastoma SH-SY5Y cells transfected with human tyrosine hydroxylase isoform 1 (SH + TH cells) were substantially more resistant to cell death induced by hydrogen peroxide and 6-hydroxydopamine when compared to wild-type SH-SY5Y cells (SH cells). SH + TH cells exhibit increased levels of dopamine (DA) compared to SH cells. Incubation with hydrogen peroxide or 6-hydroxydopamine (10-100microM) for 24 h caused a significant reduction in cell viability and increased apoptosis in both cell types. However, these effects were significantly reduced in the SH + TH cells when compared to the SH cells. The SH + TH cells showed an improved ability to detoxify peroxide, which correlated with an increase in glutathione peroxidase and glutathione reductase activities, while catalase activity was unchanged. Our data suggest that a preconditioning-like mechanism linked to higher DA levels increased the resistance of SH + TH cells against oxidative insults, which is at least in part related to an augmentation in the activity of glutathione-related antioxidant enzymes. PMID- 19815692 TI - Semiparametric estimation of the average causal effect of treatment on an outcome measured after a postrandomization event, with missing outcome data. AB - In the past decade, several principal stratification-based statistical methods have been developed for testing and estimation of a treatment effect on an outcome measured after a postrandomization event. Two examples are the evaluation of the effect of a cancer treatment on quality of life in subjects who remain alive and the evaluation of the effect of an HIV vaccine on viral load in subjects who acquire HIV infection. However, in general the developed methods have not addressed the issue of missing outcome data, and hence their validity relies on a missing completely at random (MCAR) assumption. Because in many applications the MCAR assumption is untenable, while a missing at random (MAR) assumption is defensible, we extend the semiparametric likelihood sensitivity analysis approach of Gilbert and others (2003) and Jemiai and Rotnitzky (2005) to allow the outcome to be MAR. We combine these methods with the robust likelihood based method of Little and An (2004) for handling MAR data to provide semiparametric estimation of the average causal effect of treatment on the outcome. The new method, which does not require a monotonicity assumption, is evaluated in a simulation study and is applied to data from the first HIV vaccine efficacy trial. PMID- 19815693 TI - Evaluation of pre-analytical variables in the quantification of dengue virus by real-time polymerase chain reaction. AB - An accurate molecular diagnosis for viral pathogens is highly dependent on pre analytical procedures. The efficiencies of two viral RNA extraction methods (liquid phase partition and silica-based adsorption chromatography) and the effects of handling and storage on the stability of RNA isolated from dengue virus (DENV) were studied. Viral RNA extracted from spiked sera or clinical samples characterized with DENV infection were quantified by TaqMan real-time PCR. The presence of high serum proteins severely affected the recovery of DENV RNA by the liquid phase partition, but not the silica-based method. The recovery with Trizol liquid phase partition method was significantly improved by a concomitant addition of a co-precipitant and the reduction of sera proteins, resulting in recoveries similar to that of the silica-based methods. Repeated freeze-thaw cycles did not affect the recovery of viral RNA. While intact DENV was found to be stable in serum for up to 2 hour at 25 degrees C, recovery of viral RNA from sera stored in the lysis/binding buffer was stable for up to 5 days. These data indicate that the choice of viral RNA extraction methods, the conditions for handling, and storing of clinical sera critically affect the quantification of viral nucleic acid from clinical samples. This will impact the accuracy and reproducibility of DENV diagnosis by PCR-based assays. PMID- 19815694 TI - A multicenter blinded study to evaluate KRAS mutation testing methodologies in the clinical setting. AB - Evidence that activating mutations of the KRAS oncogene abolish the response to anti-epidermal growth factor receptor therapy has revolutionized the treatment of advanced colorectal cancer. This has resulted in the urgent demand for KRAS mutation testing in the clinical setting to aid choice of therapy. The aim of this study was to evaluate six different KRAS mutation detection methodologies on two series of primary colorectal cancer samples. Two series of 80 frozen and 74 formalin-fixed paraffin-embedded tissue samples were sourced and DNA was extracted at a central site before distribution to seven different testing sites. KRAS mutations in codons 12 and 13 were assessed by using single strand conformation polymorphism analysis, pyrosequencing, high resolution melting analysis, dideoxy sequencing, or the commercially available TIB Molbiol (Berlin, Germany) or DxS Diagnostic Innovations (Manchester, UK) kits. In frozen tissue samples, concordance in KRAS status (defined as consensus in at least five assays) was observed in 66/80 (83%) cases. In paraffin tissue, concordance was 46/74 (63%) if all assays were considered or 71/74 (96%) using the five best performing assays. These results demonstrate that a variety of detection methodologies are suitable and provide comparable results for KRAS mutation analysis of clinical samples. PMID- 19815695 TI - Development of genomic DNA reference materials for genetic testing of disorders common in people of ashkenazi jewish descent. AB - Many recessive genetic disorders are found at a higher incidence in people of Ashkenazi Jewish (AJ) descent than in the general population. The American College of Medical Genetics and the American College of Obstetricians and Gynecologists have recommended that individuals of AJ descent undergo carrier screening for Tay Sachs disease, Canavan disease, familial dysautonomia, mucolipidosis IV, Niemann-Pick disease type A, Fanconi anemia type C, Bloom syndrome, and Gaucher disease. Although these recommendations have led to increased test volumes and number of laboratories offering AJ screening, well characterized genomic reference materials are not publicly available. The Centers for Disease Control and Prevention-based Genetic Testing Reference Materials Coordination Program, in collaboration with members of the genetic testing community and Coriell Cell Repositories, have developed a panel of characterized genomic reference materials for AJ genetic testing. DNA from 31 cell lines, representing many of the common alleles for Tay Sachs disease, Canavan disease, familial dysautonomia, mucolipidosis IV, Niemann-Pick disease type A, Fanconi anemia type C, Bloom syndrome, Gaucher disease, and glycogen storage disease, was prepared by the Repository and tested in six clinical laboratories using three different PCR-based assay platforms. A total of 33 disease alleles was assayed and 25 different alleles were identified. These characterized materials are publicly available from Coriell and may be used for quality control, proficiency testing, test development, and research. PMID- 19815696 TI - Ultrasensitive detection of KRAS2 mutations in bile and serum from patients with biliary tract carcinoma using LigAmp technology. AB - Patients with biliary tract carcinoma have a poor prognosis. Early detection efforts are urgently needed to ameliorate the dismal prognosis for these patients. Mutations of the KRAS2 gene are one of the most common genetic aberrations in this cancer. In this study, we used LigAmp, an ultrasensitive technology for detecting point mutations, to analyze KRAS2 mutations in patients with a variety of neoplastic and non-neoplastic pancreatobiliary diseases. DNA was isolated from 64 samples, including 44 bile samples and 20 serum samples. Oligonucleotides specific for KRAS2 G35A (GAT, G12D), G35T (GTT, G12V), and G34A (AGT, G12S) mutations were used. KRAS2 mutations were detected in 14 of 16 (87.5%) neoplastic bile samples and in 9 of 28 (32.1%) non-neoplastic bile samples. However, the mutation levels were significantly lower in the non neoplastic bile (median = 0.4%) compared with those in the neoplastic bile (median = 5.1%). KRAS2 mutations were also detected in 9 of 11 (81.8%) serum samples from patients with biliary tract carcinoma, which was further confirmed by cloning BstN1-refractory PCR products and DNA sequencing. However, KRAS2 mutations were not present in the sera from eight patients with benign pancreatobiliary diseases. These data demonstrate that KRAS2 mutations are detectable in both bile and serum using LigAmp. This technology has the potential for early biliary tract carcinoma detection and possibly for residual disease monitoring post-therapy. PMID- 19815697 TI - Molecular genotyping of hydatidiform moles: analytic validation of a multiplex short tandem repeat assay. AB - Distinction of hydatidiform moles from non-molar (NM) specimens, as well as their subclassification as complete (CHM) versus partial hydatidiform moles (PHM), is important for clinical management and accurate risk assessment for persistent gestational trophoblastic disease. Because diagnosis of hydatidiform moles based solely on morphology suffers from poor interobserver reproducibility, a variety of ancillary techniques have been developed to improve diagnosis. Immunohistochemical assessment of the paternally imprinted, maternally expressed p57 gene can identify CHMs (androgenetic diploidy) by their lack of p57 expression, but cannot distinguish PHMs (diandric monogynic triploidy) from NMs (biparental diploidy). Short tandem repeat genotyping can identify the parental source of polymorphic alleles and thus discern androgenetic diploidy, diandric triploidy, and biparental diploidy, which allows for specific diagnosis of CHMs, PHMs, and NMs, respectively. In this study, a retrospectively collected set of morphologically typical CHMs (n = 8), PHMs (n = 10), and NMs (n = 12) was subjected to an analytic validation study of both short tandem repeat genotyping and p57 immunohistochemistry. Several technical and biological problems resulted in data that were difficult to interpret. To avoid these pitfalls, we have developed an algorithm with quantitative guidelines for the interpretation of short tandem repeat genotyping data. PMID- 19815698 TI - Subclinical ADHD, stress, and coping in romantic relationships of university students. AB - OBJECTIVE: To examine how the subclinical symptoms of adult ADHD and those of oppositional-defiant disorder (ODD) affect relationship satisfaction and stress and to determine whether different patterns of coping strategies emerge when undergraduates have symptoms of one or both disorders. METHOD: Participants (N = 497) complete self-report surveys assessing ADHD and ODD symptoms, relationship satisfaction, stress, and coping strategies used in their intimate relationships. RESULTS: ADHD and ODD symptoms are significantly related to a number of stressors and different patterns of coping strategies. University students with symptoms of both ADHD and ODD display a different pattern of stressors and different patterns of coping than those with symptoms of ADHD only. CONCLUSION: Screenings for both ADHD and ODD and the provision of services that offer students more constructive coping alternatives to deal with stress in their intimate relationships are needed on university campuses. PMID- 19815699 TI - A verbal planning impairment in adult ADHD indexed by script generation tasks. AB - OBJECTIVE: Though juvenile and adult ADHD cases are well known to have a nonverbal planning impairment, a verbal-planning impairment has been demonstrated only in juvenile ADHD. The purpose of this investigation is to determine whether a verbal planning impairment also characterizes adult ADHD. METHODS: A cohort of 30 adult ADHD clients of a university psychological clinic are compared to 30 age , education-, gender-, and IQ-matched persons recruited from the general population who did not have ADHD. The dependent measure is a set of 6 paper/pencil 10-item script generation tasks. RESULTS: The findings reveal that the ADHD cohort was significantly impaired on the script task and the script task correlated significantly with severity of ADHD (CAARS index + WURS), whereas several neuropsychological measures of executive function (Stroop, COWA, Rey's Complex Figure, D2, CVLT, CPT-II) did not. Findings further showed that the script measure was weakly correlated with the other established neuropsychological measures of executive function (r < .46, shared variance of less than 21%). CONCLUSIONS: On the basis of the study findings, it is concluded that verbal planning measured with script generation tasks is distinctly impaired in clinically referred adult ADHD. PMID- 19815700 TI - Getting clued in: inferential processing and comprehension monitoring in boys with ADHD. AB - OBJECTIVE: The present study examines the ability of children with ADHD to make inferences and monitor ongoing understanding of texts, to shed light on their academic difficulties. METHOD: A total of 29 boys with ADHD and 41 comparison boys between the ages of 7 and 12 participated. Three tasks measure how boys create and evaluate inferences, particularly explanatory inferences, and how they monitor their understanding of story events and the connections among them. RESULTS: Boys with ADHD are less able than their comparison peers to make appropriate inferences, particularly explanatory inferences. They also have more trouble identifying text inconsistencies. CONCLUSIONS: Findings suggest that difficulties in making inferences and monitoring ongoing comprehension among children with ADHD may contribute to story comprehension problems and in turn to academic difficulties experienced by these children. Interventions specifically focusing on understanding causal connections, creating inferences, and monitoring ongoing understanding of stories need to be investigated. PMID- 19815701 TI - Control of lung epithelial growth by a nicotinic acetylcholine receptor: the other side of the coin. AB - This Commentary provides a perspective on an article in the current issue of the American Journal of Pathology by Maouche et al, which demonstrates that alpha7 nicotinic acetylcholine receptors play a key role in regulating airway regeneration by limiting basal epithelial cell proliferation. PMID- 19815702 TI - Latent cytomegalovirus infection exacerbates experimental colitis. AB - Inflammatory bowel disease (IBD) severity is positively correlated with cytomegalovirus (CMV) infection. This may reflect CMV triggering and/or exacerbating flares of IBD and/or IBD or immunosuppressive drugs administered to patients with IBD increasing susceptibility to CMV infection. Herein, we performed studies in mice to investigate these possibilities. Mice administered murine CMV (MCMV) developed signs of acute viral infection (malaise and weight loss) and had MCMV loads that were readily detected in numerous organs including the intestine. By 4 weeks, these mice manifested a "latent" infection in which MCMV levels were low but detectable by PCR. Such MCMV infection did not induce acute colitis in either wild-type mice or IL-10(-/-) mice, which are highly prone to developing colitis. However, underlying MCMV infection in an acute or latent state exacerbated the severity of colitis induced by dextran sodium sulfate (DSS). Such potentiation of DSS colitis by latent MCMV infection seemed to occur without viral reactivation. Whereas initial MCMV infection induced acute alterations in serum and intestinal cytokines, such cytokine levels returned to baseline before administration of DSS. However, the initial infection resulted in lasting elevation of antibodies to commensal bacterial antigens, suggesting that MCMV infection may have potentiated colitis via priming of the intestinal immune response to gut microbiota. Thus, underlying CMV infection can alter mucosal immunity, potentially increasing the tendency of CMV-infected hosts to develop colitis. PMID- 19815703 TI - Anti-neutrophil cytoplasmic antibody pathogenesis in small-vessel vasculitis: an update. AB - Vasculitides associated with serum positivity for anti-neutrophil cytoplasmic antibodies (ANCAs) that affect small- to medium-sized vessels are commonly known as ANCA-associated vasculitis (AAV) and include Wegener's granulomatosis, microscopic polyangiitis, and Churg-Strauss syndrome. Evidence derived from both in vitro studies and recent animal models points to a pathogenic role of ANCAs in AAV. In 2002, the first in vivo breakthrough in the pathogenesis of ANCAs showed that mouse ANCAs against myeloperoxidase (MPO) led to intrinsic pauci-immune renal vasculitis in mice. In 2004, a report using both in vitro and in vivo studies proposed that proteinase 3 (PR3)-directed autoimmunity involved the complementary peptide of PR3 (cPR3), which is encoded by the antisense strand of the PR3 gene. The last breakthrough came in October 2008 with a previously undescribed molecular explanation for the origin and development of injury in pauci-immune renal vasculitis, with potential clinical implications. This report showed that infection by fimbriated bacteria may trigger cross-reactive autoimmunity to a previously characterized ANCA antigen, lysosomal membrane protein-2, which is contained in the same vesicles that harbor MPO and PR3. Infection by fimbriated bacteria resulted in the production of autoantibodies, which activated neutrophils and killed human microvascular endothelium in vitro and caused renal vasculitis in rats. Although the evidence for a pathogenic role of ANCAs, mainly MPO-ANCAs, is striking, various questions remain unanswered. Understanding the key pathogenic mechanisms of AAV may provide a safer, more rational therapeutic approach than the traditional (ie, corticosteroids and immunosuppressants) treatment strategy. PMID- 19815704 TI - Genoproteomic mining of urothelial cancer suggests {gamma}-glutamyl hydrolase and diazepam-binding inhibitor as putative urinary markers of outcome after chemotherapy. AB - Urinary biomarkers for the detection of bladder cancer have been developed, but no similar markers exist for prediction of clinical outcomes after receiving chemotherapy. Here we evaluate an approach that combines genomic, proteomic, and therapeutic outcome datasets to identify novel putative urinary biomarkers of clinical outcome after neoadjuvant methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC). Using this method, we identified gamma-glutamyl hydrolase (GGH), emmprin, survivin, and diazepam-binding inhibitor (DBI). Interestingly, GGH is a protein associated with methotrexate resistance, whereas emmprin, survivin, and DBI had been previously identified as predictors of outcome after platinum-containing chemotherapeutic regimens when assessed on tumor tissue. Using disease-free survival as a marker for clinical outcome, we evaluated the ability of GGH, emmprin, survivin, and DBI expression in tumor tissue to stratify 27 patients treated with neoadjuvant MVAC. DBI (P = 0.046) but not GGH (P = 0.190), emmprin (P = 0.066), or survivin (P = 0.393) successfully stratified patients. When GGH was used with DBI the significance of stratification improved (P = 0.024), whereas the addition of survivin or emmprin to this latter two-gene model reduced its significance (P = 0.036 and P = 0.040, respectively). Although these predictive results were obtained on tumor tissues, the presence of GGH and DBI in urine serves as a rationale for developing them as urinary markers of clinical outcomes for patients treated with neoadjuvant MVAC. PMID- 19815705 TI - Contrasting actions of selective inhibitors of angiopoietin-1 and angiopoietin-2 on the normalization of tumor blood vessels. AB - Angiopoietin-1 (Ang1) and angiopoietin-2 (Ang2) have complex actions in angiogenesis and vascular remodeling due to their effects on Tie2 receptor signaling. Ang2 blocks Ang1-mediated activation of Tie2 in endothelial cells under certain conditions but is a Tie2 receptor agonist in others. We examined the effects of selective inhibitors of Ang1 (mL4-3) or Ang2 (L1-7[N]), alone or in combination, on the vasculature of human Colo205 tumors in mice. The Ang2 inhibitor decreased the overall abundance of tumor blood vessels by reducing tumor growth and keeping vascular density constant. After inhibition of Ang2, tumor vessels had many features of normal blood vessels (normalization), as evidenced by junctional accumulation of vascular endothelial-cadherin, junctional adhesion molecule-A, and platelet/endothelial cell adhesion molecule-1 in endothelial cells, increased pericyte coverage, reduced endothelial sprouting, and remodeling into smaller, more uniform vessels. The Ang1 inhibitor by itself had little noticeable effect on the tumor vasculature. However, when administered with the Ang2 inhibitor, the Ang1 inhibitor prevented tumor vessel normalization, but not the reduction in tumor vascularity produced by the Ang2 inhibitor. These findings are consistent with a model whereby inhibition of Ang2 leads to normalization of tumor blood vessels by permitting the unopposed action of Ang1, but decreases tumor vascularity primarily by blocking Ang2 actions. PMID- 19815706 TI - Enhancement of autophagic flux after neonatal cerebral hypoxia-ischemia and its region-specific relationship to apoptotic mechanisms. AB - The multiplicity of cell death mechanisms induced by neonatal hypoxia-ischemia makes neuroprotective treatment against neonatal asphyxia more difficult to achieve. Whereas the roles of apoptosis and necrosis in such conditions have been studied intensively, the implication of autophagic cell death has only recently been considered. Here, we used the most clinically relevant rodent model of perinatal asphyxia to investigate the involvement of autophagy in hypoxic ischemic brain injury. Seven-day-old rats underwent permanent ligation of the right common carotid artery, followed by 2 hours of hypoxia. This condition not only increased autophagosomal abundance (increase in microtubule-associated protein 1 light chain 3-11 level and punctuate labeling) but also lysosomal activities (cathepsin D, acid phosphatase, and beta-N-acetylhexosaminidase) in cortical and hippocampal CA3-damaged neurons at 6 and 24 hours, demonstrating an increase in the autophagic flux. In the cortex, this enhanced autophagy may be related to apoptosis since some neurons presenting a high level of autophagy also expressed apoptotic features, including cleaved caspase-3. On the other hand, enhanced autophagy in CA3 was associated with a more purely autophagic cell death phenotype. In striking contrast to CA3 neurons, those in CA1 presented only a minimal increase in autophagy but strong apoptotic characteristics. These results suggest a role of enhanced autophagy in delayed neuronal death after severe hypoxia-ischemia that is differentially linked to apoptosis according to the cerebral region. PMID- 19815708 TI - The stent-eluting drugs sirolimus and paclitaxel suppress healing of the endothelium by induction of autophagy. AB - Clinical studies have indicated that the stent-eluting drugs sirolimus and paclitaxel impact restenosis; however, it is still elusive how these drugs affect the vascular endothelium at the molecular and cellular levels. The purpose of this study was to determine whether sirolimus and paclitaxel induce molecular and cellular alterations in the vascular endothelium. Endothelial regrowth was assessed in human aortic endothelial cells and rat aortic endothelium. Molecular and cellular alterations were analyzed in human aortic endothelial cells by Western blot analysis, transmission electron microscopy, and immunofluorescence staining. Green fluorescent protein-LC3 mice were used to analyze autophagic endothelium. Here, we show that sirolimus and paclitaxel differentially induce self-digesting autophagy in vascular endothelial cells with changes in expression of LC3B, p53, and Bcl-2, considerably suppressing re-endothelialization and revascularization. These results suggest that phenotypic alteration in the endothelium by sirolimus or paclitaxel might affect the rates of late stent thrombosis, myocardial infarction, and mortality. PMID- 19815707 TI - Dysregulation of insulin signaling, glucose transporters, O-GlcNAcylation, and phosphorylation of tau and neurofilaments in the brain: Implication for Alzheimer's disease. AB - Recent studies have suggested a possible role of insulin dysfunction in the pathogenesis of sporadic Alzheimer's disease (AD). In AD, brain glucose metabolism is impaired, and this impairment appears to precede the pathology and clinical symptoms of the disease. However, the exact contribution of impaired insulin signaling to AD is not known. In this study, by using a nontransgenic rat model of sporadic AD generated by intracerebroventricular administration of streptozotocin, we investigated insulin signaling, glucose transporters, protein O-GlcNAcylation, and phosphorylation of tau and neurofilaments in the brain. We found impaired insulin signaling, overactivation of glycogen synthase kinase 3beta, decreased levels of major brain glucose transporters, down- regulated protein O-GlcNAcylation, increased phosphorylation of tau and neurofilaments, and decreased microtubule-binding activity of tau in the brains of streptozotocin treated rats. These results suggest that impaired brain insulin signaling may lead to overactivation of glycogen synthase kinase-3beta and down-regulation of O GlcNAcylation, which, in turn, facilitate abnormal hyperphosphorylation of tau and neurofilaments and, consequently, neurofibrillary degeneration. PMID- 19815709 TI - Activated cAMP response element binding protein is overexpressed in human mesotheliomas and inhibits apoptosis. AB - Little is known about the cellular mechanisms contributing to the development and chemoresistance of malignant mesothelioma (MM), an aggressive asbestos-associated tumor. A human mesothelial cell line (LP9/TERT-1) and isolated human pleural mesothelial cells showed rapid and protracted asbestos-induced cAMP response element binding protein (CREB1) phosphorylation, which was inhibited in LP9/TERT 1 cells by small molecule inhibitors of epidermal growth factor receptor phosphorylation and protein kinase A. Asbestos increased expression of several CREB target genes (c-FOS, EGR-1, MKP1, BCL2, and MMP13) and apoptosis, which was enhanced using small interfering CREB. Human MM tissue arrays showed elevated endogenous levels of phosphorylated nuclear CREB1 as compared with reactive mesothelial hyperplasias and normal lung tissue. Significantly increased phosphorylated CREB1 and mRNA levels of BCL2, c-FOS, MMP9, and MMP13 were also observed in MM cells in vitro, which were further augmented after addition of Doxorubicin (Dox). Small interfering CREB inhibited migration of MMs, increased apoptosis by Dox, and decreased BCL2 and BCL-xL expression, suggesting a role for these molecules in CREB-induced MM survival. These data indicate that CREB1 and its target genes are up-regulated in asbestos-exposed human mesothelial cells through an epidermal growth factor receptor/protein kinase A pathway. Since activated CREB1 also is increased endogenously in human MM and modifies migration and resistance to Dox-induced apoptosis, inhibition of CREB1 may be a new strategy for MM therapy. PMID- 19815711 TI - Nano-scaled particles of titanium dioxide convert benign mouse fibrosarcoma cells into aggressive tumor cells. AB - Nanoparticles are prevalent in both commercial and medicinal products; however, the contribution of nanomaterials to carcinogenesis remains unclear. We therefore examined the effects of nano-sized titanium dioxide (TiO(2)) on poorly tumorigenic and nonmetastatic QR-32 fibrosarcoma cells. We found that mice that were cotransplanted subcutaneously with QR-32 cells and nano-sized TiO(2), either uncoated (TiO(2)-1, hydrophilic) or coated with stearic acid (TiO(2)-2, hydrophobic), did not form tumors. However, QR-32 cells became tumorigenic after injection into sites previously implanted with TiO(2)-1, but not TiO(2)-2, and these developing tumors acquired metastatic phenotypes. No differences were observed either histologically or in inflammatory cytokine mRNA expression between TiO(2)-1 and TiO(2)-2 treatments. However, TiO(2)-2, but not TiO(2)-1, generated high levels of reactive oxygen species (ROS) in cell-free conditions. Although both TiO(2)-1 and TiO(2)-2 resulted in intracellular ROS formation, TiO(2)-2 elicited a stronger response, resulting in cytotoxicity to the QR-32 cells. Moreover, TiO(2)-2, but not TiO(2)-1, led to the development of nuclear interstices and multinucleate cells. Cells that survived the TiO(2) toxicity acquired a tumorigenic phenotype. TiO(2)-induced ROS formation and its related cell injury were inhibited by the addition of antioxidant N-acetyl-l-cysteine. These results indicate that nano-sized TiO(2) has the potential to convert benign tumor cells into malignant ones through the generation of ROS in the target cells. PMID- 19815710 TI - Strain differences in behavioral and cellular responses to perinatal hypoxia and relationships to neural stem cell survival and self-renewal: Modeling the neurovascular niche. AB - Premature infants have chronic hypoxia, resulting in cognitive and motor neurodevelopmental handicaps caused by suboptimal neural stem cell (NSC) repair/recovery in neurogenic zones (including the subventricular and the subgranular zones). Understanding the variable central nervous system repair response is crucial to identifying "at risk" infants and to increasing survival and clinical improvement of affected infants. Using mouse strains found to span the range of responsiveness to chronic hypoxia, we correlated differential NSC survival and self-renewal with differences in behavior. We found that C57BL/6 (C57) pups displayed increased hyperactivity after hypoxic insult; CD-1 NSCs exhibited increased hypoxia-induced factor 1alpha (HIF-1alpha) mRNA and protein, increased HIF-1alpha, and decreased prolyl hydroxylase domain 2 in nuclear fractions, which denotes increased transcription/translation and decreased degradation of HIF-1alpha. C57 NSCs exhibited blunted stromal-derived factor 1 induced migratory responsiveness, decreased matrix metalloproteinase-9 activity, and increased neuronal differentiation. Adult C57 mice exposed to hypoxia from P3 to P11 exhibited learning impairment and increased anxiety. These findings support the concept that behavioral differences between C57 and CD-1 mice are a consequence of differential responsiveness to hypoxic insult, leading to differences in HIF-1alpha signaling and resulting in lower NSC proliferative/migratory and higher apoptosis rates in C57 mice. Information gained from these studies will aid in design and effective use of preventive therapies in the very low birth weight infant population. PMID- 19815712 TI - Sphingosine kinase-1 associates with integrin {alpha}V{beta}3 to mediate endothelial cell survival. AB - Sphingosine kinase (SK)-1 promotes endothelial cell (EC) survival through the cell junction molecule CD31 (platelet endothelial cell adhesion molecule-1). The integrin alpha(v)beta(3) is also essential for EC survival; inhibition of alpha(v)beta(3) ligation promotes apoptosis. Herein we demonstrate that under basal conditions, SK-1, alpha(v)beta(3), and CD31 exist as a heterotrimeric complex. Under conditions that affect EC survival such as loss of contact with the extracellular matrix or growth factor activation, more of this heterotrimeric complex forms. Overexpression studies demonstrate a requirement for SK-1 phosphorylation at serine 225 for increased heterotrimeric complex formation, activation of alpha(v)beta(3), and EC survival signals, including Bcl-X and nuclear factor-kappaB pathways. Moreover, beta(3) integrin depletion confirmed the requirement for this heterotrimeric complex in SK-1-mediated EC survival. Thus, with alpha(v)beta(3) integrin being identifiable primarily on angiogenic ECs and SK-1 being highly expressed in tumors, targeting SK-1 may affect multiple survival pathways, and its inhibition may be highly efficacious in controlling pathological EC survival. PMID- 19815713 TI - Nodal lymphangiogenesis and metastasis: Role of tumor-induced lymphatic vessel activation in extramammary Paget's disease. AB - Nodal lymphangiogenesis promotes distant lymph node (LN) metastasis in experimental cancer models. However, the role of nodal lymphangiogenesis in distant metastasis and in the overall survival of cancer patients remains unknown. Therefore, we investigated mechanisms that might facilitate regional and distant LN metastasis in extramammary Paget's disease (EMPD). We retrospectively analyzed the impact of tumor-induced lymphatic vessel activation on the survival of 116 patients, the largest cohort with EMPD studied to date. Nodal lymphangiogenesis was significantly increased in metastatic, compared with tumor free, LNs (P = 0.022). Increased lymphatic invasion within regional LNs was significantly associated with distant metastasis in LN (P = 0.047) and organs (P = 0.003). Thus, invasion within regional LNs is a powerful indicator of systemic tumor spread and reduced patient survival in EMPD (P = 0.0004). Lymphatic vessels associated with tumors expressed stromal cell-derived factor-1 (SDF-1), whereas CXCR4 was expressed on invasive Paget cells undergoing epithelial-mesenchymal transition (EMT)-like process. A431 cells overexpressing Snail expressed increased levels of CXCR4 in the presence of transforming growth factor-beta1. Haptotactic migration assays confirmed that Snail-induced EMT-like process promotes tumor cell motility via the CXCR4-SDF-1 axis. Sinusoidal lymphatic endothelial cells and macrophages expressed SDF-1 in subcapsular sinuses of lymph nodes before Paget cell arrival. Our findings reveal that EMT-related features likely promote lymphatic metastasis of EMPD by activating the CXCR4-SDF-1 axis. PMID- 19815714 TI - Thymic indoleamine 2,3-dioxygenase-positive eosinophils in young children: potential role in maturation of the naive immune system. AB - Eosinophils expressing indoleamine 2, 3-dioxygenase (IDO) may contribute to T helper cell (Th)2 predominance. To characterize human thymus IDO+ eosinophil ontogeny relative to Th2 regulatory gene expression, we processed surgically obtained thymi from 22 children (age: 7 days to 12 years) for immunohistochemistry and molecular analysis, and measured cytokine and kynurenine levels in tissue homogenates. Luna+ eosinophils ( approximately 2% of total thymic cells) decreased in number with age (P = 0.02) and were IDO+. Thymic IDO immunoreactivity (P = 0.01) and kynurenine concentration (P = 0.01) decreased with age as well. In addition, constitutively-expressed interleukin (IL)-5 and IL 13 in thymus supernatants was highest in youngest children. Eosinophil numbers correlated positively with expression of the Th2 cytokines IL-5, IL-13 (r = 0.44, P = 0.002), and IL-4 (r = 0.46, P = 0.005), transcription factor signal transducer and activator of transcription-6 (r = 0.68, P = 0.001), and the chemokine receptor, CCR3 (r = 0.17, P = 0.04), but negatively with IL-17 mRNA (r = -0.57, P = 0.02) and toll-like receptor 4 expression (r = -0.74, P = 0.002). Taken together, these results suggest that functional thymic IDO+ eosinophils during human infant life may have an immunomodulatory role in Th2 immune responses. PMID- 19815715 TI - Unilateral blood flow decrease induces bilateral and symmetric responses in the immature brain. AB - The effects of hemodynamic changes in the developing brain have yet to be fully understood. The aim of this study was to explore the relationship between perturbations of the cerebral blood flow in the developing brain via unilateral hypoperfusion in P7 rats. As expected, nuclear caspase-3 (casp3) cleavage and DNA fragmentation were detected at 48 hours after stroke in the injured cortex. Surprisingly, casp3 was also cleaved in the contralateral cortex, although without cell death markers. Delayed (48 hours) casp3 cleavage without DNA fragmentation was also identified after unilateral common carotid artery occlusion, both in the hypoperfused cortex and the unaffected cortex, producing mirror images. Upstream calpain activation, caspase-2 cleavage, and mitochondrial cytochrome c release initiated casp3 cleavage, but did not produce preconditioning. The neuronal marker NeuN co-localized with cleaved casp3 in cortical layers II-III and VI and with gaba-amino butyric acid in layer III. Indeed, collateral supply was provided from the opposite side during carotid artery occlusion but not after reperfusion, and the number of cleaved casp3 positive cells significantly negatively correlated with the common carotid artery immediate reperfusion percentage. In summary, unilateral hypoperfusion, while insufficient to induce cell death, may active bilateral and symmetric casp3 in the P7 rat brain. Additionally, the opposite healthy hemisphere is altered due to the injury and thus should not be used as an internal control. PMID- 19815717 TI - Say NO to hypoperfusion! PMID- 19815716 TI - Transient activation of Wnt/{beta}-catenin signaling induces abnormal growth plate closure and articular cartilage thickening in postnatal mice. AB - Wnt/beta-catenin signaling is required for skeletal development and organization and for function of the growth plate and articular cartilage. To further clarify these roles and their possible pathophysiological importance, we created a new transgenic mouse model in which Wnt/beta-catenin signaling can be activated in cartilage for specific periods of time. These transgenic mice expressed a constitutive active form of beta-catenin fused to a modified estrogen receptor ligand-binding domain under the control of cartilage-specific collagen 11alpha2 promoter/enhancer. Transient Wnt/beta-catenin signaling activation in young adult mice by tamoxifen injections induced growth retardation and severe deformities in knee joints. Tibial and femoral growth plates displayed an excessive number of apoptotic cells and eventually underwent abnormal regression. Articular cartilage exhibited an initial acute loss of proteoglycan matrix that was followed by increases in thickness, cell density, and cell proliferation. In reciprocal studies, we found that conditional ablation of beta-catenin in postnatal mice using a Col2-CreER strategy led to hypocellularity in articular cartilage, growth plate disorganization, and a severe reduction in bone volume. Together, these data provide evidence that Wnt/beta-catenin signaling has important and distinct roles in growth plate and articular cartilage and that postnatal dysregulation of this signaling pathway causes diverse structural and functional changes in the two cartilaginous structures. PMID- 19815718 TI - In memoriam: Jere Mead (1920-2009). PMID- 19815719 TI - Effect of whole body heat stress on peripheral vasoconstriction during leg dependency. AB - The venoarteriolar response (VAR) increases vascular resistance upon increases in venous transmural pressure in cutaneous, subcutaneous, and muscle vascular beds. During orthostasis, it has been proposed that up to 45% of the increase in systemic vascular tone is due to VAR-related local mechanism(s). The objective of this project was to test the hypothesis that heat stress attenuates VAR-mediated cutaneous and whole leg vasoconstriction. During normothermic conditions, measurements of cutaneous blood flow (laser-Doppler flowmetry) and femoral artery blood flow (Doppler ultrasound) were obtained from both legs during supine and leg-dependent conditions. These measurements were repeated following a whole body heat stress (increase in internal temperature of 1.4 +/- 0.2 degrees C). Before leg dependency, cutaneous (CVC) and femoral vascular conductances (FVC) were significantly elevated in both legs during heat stress relative to normothermia (P < 0.001). During leg dependency the absolute decrease in CVC was attenuated during heat stress (P < 0.01) while the absolute decrease in FVC was unaffected (P = 0.90). When CVC and FVC data were analyzed as a relative change from their respective baseline values, heat stress significantly attenuated the magnitude of vasoconstriction due to leg dependency in the cutaneous and femoral circulations (P < 0.001 for both variables). These data suggest that an attenuated local vasoconstriction, evoked via the venoarteriolar response, may contribute to reduced blood pressure control and thus reduced orthostatic tolerance that occurs in heat-stressed individuals. PMID- 19815720 TI - Contrasting effects of simulated microgravity with and without daily -Gx gravitation on structure and function of cerebral and mesenteric small arteries in rats. AB - This study was designed to test the hypothesis that a 28-day tail suspension (SUS) could induce hypertrophy and enhanced myogenic and vasoconstrictor reactivity in middle cerebral arteries (MCAs), whereas atrophy and decreased myogenic and vasoconstrictor responses in mesenteric third-order arterioles (MSAs). Also, in addition to the functional enhancement in MCAs, structural changes in both kinds of arteries and functional decrement in MSAs could all be prevented by the intervention of daily 1-h dorsoventral (-G(x)) gravitation by restoring to standing posture. To test this hypothesis, vessel diameters to pressure alterations and nonreceptor- and receptor-mediated agonists were determined using a pressure arteriograph with a procedure to measure in vivo length and decrease hysteresis of vessel segments and longitudinal middlemost sections of vessels fixed at maximally dilated state were examined using electron microscopy and histomorphometry. Functional studies showed that 28-day tail suspended, head-down tilt (SUS) resulted in enhanced and decreased myogenic tone and vasoconstrictor responses, respectively, in MCAs and MSAs. Histomorphometric data revealed that SUS-induced hypertrophic changes in MCAs characterized by increases in thickness (T) and cross-sectional area (CSA) of the media and the number of vascular smooth-muscle-cell layers (N(CL)), whereas in MSAs, it induced decreases in medial CSA and T and N(CL). Daily 1-h -G(x) over 28 days can fully prevent these differential structural changes in both kinds of small arteries and the functional decrement in MSAs, but not the augmented myogenic tone and increased vasoreactivity in the MCAs. These findings have revealed special features of small resistance arteries during adaptation to microgravity with and without gravity-based countermeasure. PMID- 19815721 TI - Virology. A new virus for old diseases? PMID- 19815722 TI - Genotype analysis identifies the cause of the "royal disease". AB - The "royal disease," a blood disorder transmitted from Queen Victoria to European royal families, is a striking example of X-linked recessive inheritance. Although the disease is widely recognized to be a form of the blood clotting disorder hemophilia, its molecular basis has never been identified, and the royal disease is now likely extinct. We identified the likely disease-causing mutation by applying genomic methodologies (multiplex target amplification and massively parallel sequencing) to historical specimens from the Romanov branch of the royal family. The mutation occurs in F9, a gene on the X chromosome that encodes blood coagulation factor IX, and is predicted to alter RNA splicing and to lead to production of a truncated form of factor IX. Thus, the royal disease is the severe form of hemophilia, also known as hemophilia B or Christmas disease. PMID- 19815724 TI - Coupling of CO2 and ice sheet stability over major climate transitions of the last 20 million years. AB - The carbon dioxide (CO2) content of the atmosphere has varied cyclically between approximately 180 and approximately 280 parts per million by volume over the past 800,000 years, closely coupled with temperature and sea level. For earlier periods in Earth's history, the partial pressure of CO2 (pCO2) is much less certain, and the relation between pCO2 and climate remains poorly constrained. We use boron/calcium ratios in foraminifera to estimate pCO2 during major climate transitions of the past 20 million years. During the Middle Miocene, when temperatures were approximately 3 degrees to 6 degrees C warmer and sea level was 25 to 40 meters higher than at present, pCO2 appears to have been similar to modern levels. Decreases in pCO(2) were apparently synchronous with major episodes of glacial expansion during the Middle Miocene (approximately 14 to 10 million years ago) and Late Pliocene (approximately 3.3 to 2.4 million years ago). PMID- 19815723 TI - Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome. AB - Chronic fatigue syndrome (CFS) is a debilitating disease of unknown etiology that is estimated to affect 17 million people worldwide. Studying peripheral blood mononuclear cells (PBMCs) from CFS patients, we identified DNA from a human gammaretrovirus, xenotropic murine leukemia virus-related virus (XMRV), in 68 of 101 patients (67%) as compared to 8 of 218 (3.7%) healthy controls. Cell culture experiments revealed that patient-derived XMRV is infectious and that both cell associated and cell-free transmission of the virus are possible. Secondary viral infections were established in uninfected primary lymphocytes and indicator cell lines after their exposure to activated PBMCs, B cells, T cells, or plasma derived from CFS patients. These findings raise the possibility that XMRV may be a contributing factor in the pathogenesis of CFS. PMID- 19815725 TI - Microsecond simulations of spontaneous methane hydrate nucleation and growth. AB - Despite the industrial implications and worldwide abundance of gas hydrates, the formation mechanism of these compounds remains poorly understood. We report direct molecular dynamics simulations of the spontaneous nucleation and growth of methane hydrate. The multiple-microsecond trajectories offer detailed insight into the process of hydrate nucleation. Cooperative organization is observed to lead to methane adsorption onto planar faces of water and the fluctuating formation and dissociation of early hydrate cages. The early cages are mostly face-sharing partial small cages, favoring structure II; however, larger cages subsequently appear as a result of steric constraints and thermodynamic preference for the structure I phase. The resulting structure after nucleation and growth is a combination of the two dominant types of hydrate crystals (structure I and structure II), which are linked by uncommon 5(12)6(3) cages that facilitate structure coexistence without an energetically unfavorable interface. PMID- 19815726 TI - Axonal/glial upregulation of EphB/ephrin-B signaling in mouse experimental ocular hypertension. AB - PURPOSE: To use a laser-induced ocular hypertension (LIOH) mouse model to examine the optic nerve head (ONH) expression of EphB/ephrin-B, previously shown to be upregulated in glaucomatous DBA/2J mice. To relate ephrin-B reverse signaling with states of axonal response to disease. METHODS: LIOH was induced unilaterally in CD-1 mice by laser photocoagulation of limbal and episcleral veins. Intraocular pressure (IOP) was measured with a tonometer. EphB/ephrin-B mRNA expression was assessed by in situ hybridization on eyecup cryosections and real time PCR. Cell specific markers were used to identify the cellular origin of EphB/ephrin-B expression. Activation of ephrin-B signaling was investigated with a phosphospecific antibody on cryosections and retinal whole-mounts. RESULTS: Upregulation of EphB/ephrin-B expression occurred early within a day of IOP elevation. A transient increase of phosphorylation-dependent ephrin-B (pEB) reverse signaling was observed in ONH axons, microglia, and some astrocytes. Morphologically unaffected retinal ganglion cell (RGC) axons differed from axons with reactive aberrant trajectories by exhibiting increased pEB activation, whereas pEB levels in morphologically affected axons were comparable to those of controls. CONCLUSIONS: An Eph-ephrin signaling network is activated at the ONH after LIOH in CD-1 mice, either before or coincident with the initial morphologic signs of RGC axon damage reported previously. Of note, ephrin-B reverse signaling was transiently upregulated in RGC axons at the ONH early in their response to IOP elevation but was downregulated in axons that had been damaged by glaucomatous injury and exhibited aberrant trajectories. Ephrin-B reverse signaling may mark RGC axons for damage or confer a protective advantage against injury. PMID- 19815727 TI - Vision-related quality of life and visual function after vitrectomy for various vitreoretinal disorders. AB - PURPOSE: To investigate vision-related quality of life (VR-QOL) in patients undergoing vitrectomy for various vitreoretinal disorders and to evaluate the relationship between VR-QOL and visual function. METHODS: The study included 100 normal control subjects and 299 patients with various vitreoretinal disorders including proliferative diabetic retinopathy (PDR), diabetic macular edema (DME), branch retinal vein occlusion (BRVO), central retinal vein occlusion (CRVO), macular hole (MH), epiretinal membrane (ERM), and rhegmatogenous retinal detachment (RD). The 25-item National Eye Institute Visual Function Questionnaire (VFQ-25) was answered by the patients with vitreoretinal disorders before and 3 months after pars plana vitrectomy, as well as by the normal control subjects. Clinical data were collected, including visual acuity, contrast sensitivity, and severity of metamorphopsia. RESULTS: Vitrectomy significantly improved the VFQ-25 composite score in all vitreoretinal disorders. Preoperative VFQ-25 composite scores in MH and ERM were significantly higher than those in PDR, DME, and BRVO. Postoperative VFQ-25 composite scores were significantly higher in MH, ERM, and RD than in PDR, DME, BRVO, and CRVO. A greater improvement in the VFQ-25 composite score was observed in ERM than in DME. Multiple regression analysis revealed that changes in contrast sensitivity had a significant correlation with changes in the VFQ-25 composite score in PDR and DME. Changes in metamorphopsia were significantly associated with changes in the VFQ-25 composite score in MH and ERM. CONCLUSIONS: Vitrectomy significantly improved VR-QOL in various vitreoretinal disorders. The largest improvement in VR-QOL was observed in ERM and smallest improvement in DME. The visual function parameters associated with VR-QOL are different depending on vitreoretinal disorders. PMID- 19815728 TI - Differential relative sulfation of Keratan sulfate glycosaminoglycan in the chick cornea during embryonic development. AB - PURPOSE: To investigate structural remodeling of the developing corneal stroma concomitant with changing sulfation patterns of keratan sulfate (KS) glycosaminoglycan (GAG) epitopes during embryogenesis and the onset of corneal transparency. METHODS: Developing chick corneas were obtained from embryonic day (E)12 to E18 of incubation. Extracellular matrix composition and collagen fibril spacing were evaluated by synchrotron x-ray diffraction, hydroxyproline assay, ELISA (with antibodies against lesser and more highly sulfated KS), and transmission electron microscopy with specific proteoglycan staining. RESULTS: A significant relative increase in highly sulfated KS epitope labeling occurred with respect to hydroxyproline content in the final week of chick development, as mean collagen interfibrillar distance decreased. Small KS PG filaments increased in frequency with development and were predominantly fibril associated. CONCLUSIONS: The accumulation of highly sulfated KS during the E12 to E18 timeframe could serve to fine tune local matrix hydration and collagen fibril spacing during corneal growth, as gross dehydration and compaction of the stroma progress through the action of the nascent endothelial pump. PMID- 19815730 TI - Assessment of the integrin alpha5beta1 antagonist JSM6427 in proliferative vitreoretinopathy using in vitro assays and a rabbit model of retinal detachment. AB - PURPOSE: To explore the role of integrin alpha5beta1 in proliferative vitreoretinopathy (PVR) pathogenesis by evaluating the expression alpha5beta1 on ARPE-19 cells and patient proliferative membranes, quantifying the inhibitory effects of JSM6427 (a small molecule alpha5beta1 inhibitor) on ARPE-19 cell adhesion and migration, and assessing the therapeutic potential of JSM6427 in a rabbit retinal detachment model. METHODS: Expression of alpha5beta1 was evaluated on activated ARPE-19 cells by flow cytometry and on PVR membranes by immunohistochemistry. ARPE-19 cells were used in fibronectin-dependent adhesion and migration assays with various concentrations of JSM6427; IC(50) was calculated. In the rabbit model, eyes were intravitreally injected with vehicle or JSM6427 on day 0 or 1 after retinal detachment; BrdU was administered intravitreally on day 3, and retinal tissues were harvested on day 3 (4 hours later) or 7. Retinal scarring, cellular proliferation, and inflammatory responses were quantified, and retinal morphology was analyzed in retinal sections. RESULTS: Activated ARPE-19 cells and PVR membranes expressed high levels of alpha5beta1; expression was low in control eyes. JSM6427 provided a dose dependent blockade of ARPE-19 cell adhesion to fibronectin (IC(50), 7.1 +/- 2.5 microM) and inhibition of migration (IC(50), 6.0 +/- 4.5 microM). In the rabbit model, intravitreal injection of JSM6427 provided significant inhibition of proliferation of retinal cells (Muller cells, microglia, and macrophages) on days 3 and 7 after detachment and inhibition of inflammatory response and retinal scarring on day 7 after detachment. CONCLUSIONS: JSM6427 is a promising treatment for PVR, with data suggesting that inhibition of alpha5beta1-fibronectin interactions addresses multiple pathways involving retinal pigment epithelial, glial, and inflammatory cells. PMID- 19815729 TI - Growth factor regulation of corneal keratocyte differentiation and migration in compressed collagen matrices. AB - PURPOSE: To evaluate a novel 3D culture model of the corneal stroma and apply it to investigate how key wound-healing growth factors regulate the mechanics of corneal keratocyte migration. METHODS: Rabbit corneal keratocytes were seeded within collagen matrices that were compacted using external compression. Six millimeter-diameter buttons were then incubated in media supplemented with 10% FBS, TGFbeta1, TGFbeta2, platelet-derived growth factor (PDGF), or no growth factor (control). After 1, 3, or 7 days, matrices were labeled with phalloidin and a nucleic acid dye, and were imaged using laser confocal microscopy. To study cell migration, buttons were nested within acellular uncompressed outer collagen matrices before growth factor stimulation. RESULTS: Corneal keratocytes in basal media within compressed matrices had a broad, convoluted cell body and thin dendritic processes. In contrast, cells in 10% FBS developed a bipolar fibroblastic morphology. Treatment with TGFbeta induced the formation of stress fibers expressing alpha-smooth muscle actin, suggesting myofibroblast transformation. PDGF induced keratocyte elongation without inducing stress fiber formation. Both 10% FBS and PDGF stimulated significant keratocyte migration through the uncompressed outer matrix, but 10% FBS produced more cell-induced collagen matrix reorganization. TGFbeta induced the smallest increase in migration and the greatest matrix reorganization. CONCLUSIONS: Corneal keratocytes are able to differentiate normally and respond to growth factors within compressed collagen matrices, which provide a high-stiffness, 3D environment, similar to native stromal tissue. In addition, nesting these matrices provides a unique platform for investigating the mechanics of keratocyte migration after exposure to specific wound-healing cytokines. PMID- 19815731 TI - Detection of glaucoma progression with stratus OCT retinal nerve fiber layer, optic nerve head, and macular thickness measurements. AB - PURPOSE: To evaluate and compare the ability of optical coherence tomography (OCT) retinal nerve fiber layer (RNFL), optic nerve head, and macular thickness parameters to detect progressive structural damage in glaucoma. METHODS: This observational cohort study included 253 eyes of 253 patients. Images were obtained annually with the Stratus OCT (Carl Zeiss Meditec, Inc., Dublin, CA) along with optic disc stereophotographs and standard automated perimetry (SAP) visual fields. The median follow-up time was 4.01 years. Progression was determined by the Guided Progression Analysis software for SAP (Carl Zeiss Meditec, Inc.) and by masked assessment of optic disc stereophotographs performed by expert graders. Random coefficient models and receiver operating characteristic (ROC) curves were used to evaluate the relationship between change in Stratus OCT parameters over time and progression as determined by SAP and/or stereophotographs. RESULTS: From the 253 eyes, 31 (13%) showed progression over time by stereophotographs and/or SAP. Mean rates of change in average RNFL thickness were significantly higher for progressors compared with nonprogressors (-0.72 mum/y vs. 0.14 mum/y; P = 0.004), with sensitivity of 77% for specificity of 80%. RNFL parameters performed significantly better than ONH and macular thickness measurements in discriminating progressors from nonprogressors. The parameters with the largest ROC curve areas for each scanning area were inferior RNFL thickness (0.84), cup area (0.66), and inferior inner macula thickness (0.64). CONCLUSIONS: Stratus OCT RNFL parameters discriminated between eyes progressing by visual fields or optic disc photographs and eyes that remained stable by these methods and performed significantly better than ONH and macular thickness parameters in detecting change over time. PMID- 19815733 TI - Optical functional properties of the Boston Keratoprosthesis. AB - PURPOSE: To evaluate the optical characteristics of the Boston Keratoprosthesis (KPro), identify glare sources, evaluate possible glare control, and examine the benefit of implantation when the fellow eye has normal vision. METHODS: Computed and optical-bench-measured point spread function (PSF) and glare sources were compared. A translucent plastic cornea was used to determine the impact of glare caused by scatter in the cornea and its control with a dark-iris tinted contact lens. The effect of glare in implanted eyes was measured with a brightness acuity test (BAT), with and without the dark-iris contact lens. Computed and measured visual fields were compared. Stereopsis was measured in patients with an intact fellow eye. RESULTS: Computed and measured modulation transfer functions for the KPro were found to be very close to the diffraction limit. Both the model-eye measurements and patients' BAT glare responses identified that the hazy corneal graft surrounding the KPro is the main source of glare and can be controlled with a dark-iris contact lens. The lid effectively blocks the light that would be scattered in the hazy cornea of patients in whom the type II KPro was implanted. An intact fellow eye remains the dominant eye, with better acuity, and the KPro eye supports only minimal stereo ability and does not expand the binocular visual field. CONCLUSIONS: Glare can be reduced significantly with the use of a contact lens with a dark iris. Implanting the KPro in a patient whose fellow eye has normal or near normal vision does not seem to improve visual function. PMID- 19815732 TI - Protection of retinal ganglion cells by caspase substrate-binding peptide IQACRG from N-methyl-D-aspartate receptor-mediated excitotoxicity. AB - PURPOSE: This study investigated whether the enzymatically inactive caspase mimetic IQACRG protects rat retinal ganglion cells (RGCs) from excitotoxic insults. Minimally invasive delivery of the peptide to the retina was explored, and the mechanisms of neuroprotection were elucidated. METHODS: IQACRG was linked to penetratin (P-IQACRG) to facilitate cellular uptake. RGC labeling by biotinylated-P-IQACRG delivered via intravitreal or subconjunctival injection was demonstrated by avidin-biotin chemistry. The authors used histologic and electrophysiological measures to evaluate the neuroprotective potential of P IQACRG against RGC death induced by N-methyl-D-aspartate (NMDA) in vitro and in vivo. In addition, they monitored activity of an enzyme that is downstream of caspase-1, matrix metalloproteinase-9 (MMP-9), and protein levels of the caspase 3/7 substrate, myocyte enhancer factor 2C (MEF2C), to determine the effectiveness of IQACRG in blocking excessive caspase activity. RESULTS: IQACRG significantly reduced NMDA-induced RGC death in culture and in vivo. Ex vivo electrophysiological recording of the retina on multielectrode arrays demonstrated functional rescue of RGCs by IQACRG. The authors also found that delivery of IQACRG to the retina inhibited NMDA-triggered MMP-9 activity and prevented cleavage of MEF2C protein that would otherwise have been engendered by caspase activation preceding RGC death. Strikingly, subconjunctival injection of P-IQACRG was very effective in preventing NMDA-induced RGC death in vivo. CONCLUSIONS: These data demonstrate that IQACRG protects RGCs from excitotoxicity in vitro and in vivo. The positive results with subconjunctival administration of P-IQACRG suggest that in the future this treatment may be useful clinically in diseases such as glaucoma and retinal ischemia. PMID- 19815734 TI - Donor and recipient endothelial cell population of the transplanted human cornea: a two-dimensional imaging study. AB - Purpose. To elucidate the pattern of donor and recipient endothelial cell populations in transplanted human corneas and determine the degree to which donor endothelial cells survive in the graft. Methods. Thirty-six corneal grafts were collected from recipients of opposite sex to the donor, at the time of retransplantation for various indications. Cells from the endothelial side of the grafts were harvested, preserving their relative location on the endothelium. Fluorescence in situ hybridization of the sex chromosomes enabled each cell to be identified as donor- or recipient-derived. Images of the graft endothelium were assembled, to depict the pattern of cell population of the graft, and the proportion of donor cells present was estimated. Results. Endothelial cells of donor origin were found in 26 of 36 grafts (72.2%)-in one case, up to 26 years after transplantation. The proportion of donor endothelium ranged from 2% to 99%; however, there was no significant correlation of this proportion with postoperative time (P = 0.19). The mean annual rate of donor cell loss correlated negatively with the time to graft failure by endothelial decompensation (P = 0.002). Endothelial images indicated a highly variable pattern of recipient cell repopulation of the graft. A tendency toward donor cell retention in transparent, successful grafts was noted; however, this feature alone was not a reliable indicator of long-term graft transparency. Conclusions. Two-dimensional imaging of the corneal graft endothelium revealed a variable pattern and extent of donor and recipient cell population, indicating the highly dynamic nature of the corneal endothelium after transplantation. PMID- 19815735 TI - Potentiation of femtosecond laser intratissue refractive index shaping (IRIS) in the living cornea with sodium fluorescein. AB - PURPOSE: To assess the effectiveness of intratissue refractive index shaping (IRIS) in living corneas and test the hypothesis that it can be enhanced by increasing the two-photon absorption (TPA) of the tissue. METHODS: Three corneas were removed from adult cats and cut into six pieces, which were placed in preservative (Optisol-GS; Bausch & Lomb, Inc., Irvine, CA) containing 0%, 0.25%, 1%, 1.5%, or 2.5% sodium fluorescein (Na-Fl). An 800-nm Ti:Sapphire femtosecond laser with a 100-fs pulse duration and 80-MHz repetition rate was used to perform IRIS in each piece, creating several refractive index (RI) modification lines at different speeds (between 0.1 and 5 mm/s). The lines were 1 mum wide, 10 microm apart, and approximately 150 microm below the tissue surface. The RI change of each grating was measured using calibrated, differential interference contrast microscopy. TUNEL staining was performed to assess whether IRIS or Na-Fl doping causes cell death. RESULTS: Scanning at 0.1 mm/s changed the RI of undoped, living corneas by 0.005. In doped corneas, RI changes between 0.01 and 0.02 were reliably achieved with higher scanning speeds. The magnitude of RI changes attained was directly proportional to Na-Fl doping concentration and inversely proportional to the scanning speed used to create the gratings. CONCLUSIONS: IRIS can be efficiently performed in living corneal tissue. Increasing the TPA of the tissue with Na-Fl increased both the scanning speeds and the magnitude of RI changes in a dose-dependent manner. Ongoing studies are exploring the use of IRIS to alter the optical properties of corneal tissue in situ, over an extended period. PMID- 19815736 TI - Endothelial nitric oxide synthase gene variants and primary open-angle glaucoma: interactions with sex and postmenopausal hormone use. AB - PURPOSE: To evaluate the association between the nitric oxide synthase gene (NOS3) variants and primary open-angle glaucoma (POAG). METHODS: Two functional single-nucleotide polymorphisms (SNPs) (T-786C: rs2070744; Glu298Asp: rs1799983) and three tagging SNPs (rs7830, rs3918188, and rs1800779) were evaluated in a nested case-control study from the Nurses' Health Study (1980-2002) and the Health Professionals' Follow-up Study (1986-2002). Participants were aged >or=40 years and Caucasian. Included were 527 incident cases and 1543 controls, matched by cohort, age, and eye examination at the matched cases' diagnosis dates. Cohort specific relative risks (RR) were estimated by using multivariable conditional logistic regression and were pooled with meta-analysis. RESULTS: No NOS3 polymorphism was significantly associated with overall POAG. For high-tension POAG (HTPOAG), rs3918188 was significantly inversely associated among the women (AA versus CC genotype: RR = 0.48; 95% CI, 0.28-0.82) but not among the men (P heterogeneity by sex = 0.02). The minor alleles of T -786C and rs1800779 showed positive association with high-tension POAG (P-trend < 0.02) in the women only, but P-heterogeneity was not significant. In the women, four of the five NOS3 SNPs showed significant interactions with postmenopausal hormone (PMH) use in relation to HTPOAG: for example, among the women with the TT genotype in T -786C, PMH use was inversely associated (RR = 0.41; 95% CI, 0.22-0.76), but among carriers of the minor allele, use of PMH was not associated. CONCLUSIONS: Interactions were observed between NOS3 SNPs and female sex and postmenopausal hormone use in the women in relation to HTPOAG. These findings should be confirmed in different racial/ethnic groups. PMID- 19815738 TI - Laser-induced ocular hypertension in albino CD-1 mice. AB - PURPOSE: To establish a laser-induced model of ocular hypertension (LIOH) in albino CD-1 mice and to characterize the sequence of pathologic events triggered by intraocular pressure (IOP) elevation. METHODS: LIOH was induced unilaterally in CD-1 mice by laser photocoagulation of limbal and episcleral veins 270 degrees to 300 degrees circumferentially, sparing the nasal aspect and the long ciliary arteries. IOP was measured with a rebound tonometer. Hematoxylin and eosin stained plastic sections were used for morphometric analysis of retinal layers, and retinal whole-mounts were immunostained with anti-Brn-3b to quantify retinal ganglion cell (RGC) gene expression ion and density. Axonal and myelin morphologies were characterized using appropriate antibodies, and axon counts were obtained from paraphenylenediamine-stained optic nerve sections. RESULTS: LIOH resulted in IOP doubling within 4 hours after laser treatment, which returned to normal by 7 days. Axon degenerative changes, reactive plasticity, and aberrant regrowth were detected at the optic nerve head (ONH) as early as 4 days after treatment. By 7 days, axon number was significantly reduced in the myelinated optic nerve, with concurrent signs of myelin degradation. At 14 days, Brn-3b(+) RGC density was reduced, with neuronal loss confined to the RGC layer and no apparent effects on other retinal layers. CONCLUSIONS: Laser photocoagulation of limbal and episcleral veins induces transient ocular hypertension in albino CD-1 mice. The ensuing retinal and optic nerve pathologic events recapitulated key features of glaucoma and placed ONH RGC axon responses as an early manifestation of damage. LIOH in albino mice may be useful as a mouse model to examine mechanisms of RGC and axon glaucomatous injury. PMID- 19815737 TI - Morphology and accommodative function of the vitreous zonule in human and monkey eyes. AB - PURPOSE: To explore the attachments of the posterior zonule and vitreous in relation to accommodation and presbyopia in monkeys and humans. METHODS: Novel scanning electron microscopy (SEM) and ultrasound biomicroscopy (UBM) techniques were used to visualize the anterior, intermediate, and posterior vitreous zonule and their connections to the ciliary body, vitreous membrane, lens capsule, and ora serrata, and to characterize their age-related changes and correlate them with loss of accommodative forward movement of the ciliary body. alpha Chymotrypsin was used focally to lyse the vitreous zonule and determine the effect on movement of the accommodative apparatus in monkeys. RESULTS: The vitreous attached to the peripheral lens capsule and the ora serrata directly. The pars plana zonule and the posterior tines of the anterior zonule were separated from the vitreous membrane except for strategically placed attachments, collectively termed the vitreous zonule, that may modulate and smooth the forward and backward movements of the entire system. Age-dependent changes in these relationships correlated significantly with loss of accommodative amplitude. Lysis of the intermediate vitreous zonule partially restored accommodative movement. CONCLUSIONS: The vitreous zonule system may help to smoothly translate to the lens the driving forces of accommodation and disaccommodation generated by the ciliary muscle, while maintaining visual focus and protecting the lens capsule and ora serrata from acute tractional forces. Stiffening of the vitreous zonular system may contribute to age-related loss of accommodation and offer a therapeutic target for presbyopia. PMID- 19815740 TI - The art of translation. PMID- 19815739 TI - Association between high tear epidermal growth factor levels and corneal subepithelial fibrosis in dry eye conditions. AB - PURPOSE: To compare tear epidermal growth factor (EGF) concentration in dry eye (DE) conditions and determine correlations between EGF levels and severity of symptoms and ocular surface signs. METHODS: In this prospective case-control study, 35 patients with DE, including subgroups with meibomian gland disease (MGD), Sjogren's syndrome (SS) aqueous tear deficiency, or neurotrophic keratopathy (NK), and 17 asymptomatic control subjects were evaluated. Symptoms, Schirmer test, fluorescein clearance test (FCT), EGF concentration, dye staining, and the presence of corneal subepithelial fibrosis and meibomian gland (MG) orifice metaplasia were recorded. Tear EGF and the severity of irritation and ocular surface signs were correlated. RESULTS: Tear EGF was higher in MGD than in the control (P = 0.03) and was lower in SS than in the control (P < 0.0001; MGD (P < 0.05) and NK (P < 0.01) groups. The DE subgroup with results in the FCT > 3 and Schirmer 1 >or= 8 had higher EGF levels than the group with FCT > 3 and Schirmer 1 < 8 and both groups with good tear clearance (P < 0.01). Tear EGF levels correlated inversely with conjunctival (r = -0.49, P = 0.0032) and corneal (r = -0.39, P = 0.022) dye staining and positively with MG orifice metaplasia (r = 0.36, P = 0.03) and corneal subepithelial fibrosis (r = 0.5, P = 0.0006). CONCLUSIONS: Tear EGF concentration was increased in eyes with MGD, corneal subepithelial fibrosis, and MG orifice metaplasia. Elevated tear EGF may promote development of corneal subepithelial fibrosis and lid margin changes. PMID- 19815742 TI - Physics Nobel. Digital imaging, communication advances honored. PMID- 19815741 TI - Physiology Nobel. U.S. Researchers recognized for work on telomeres. PMID- 19815743 TI - Newsmaker interview. Francis Collins: looking beyond the funding deluge. Interview by Jocelyn Kaiser. PMID- 19815745 TI - Virology. Chronic fatigue and prostate cancer: a retroviral connection? PMID- 19815746 TI - U.S. research policy. Agricultural science gets more money, new faces. PMID- 19815748 TI - Climate change. Both of the world's ice sheets may be shrinking faster and faster. PMID- 19815749 TI - Cancer research. Looking for a target on every tumor. PMID- 19815750 TI - National labs. For a famous physics laboratory, a quick and painful rebirth. PMID- 19815751 TI - Paleoanthropology. New work may complicate history of Neandertals and H. sapiens. PMID- 19815752 TI - News story on Italy's MIT disappoints. PMID- 19815753 TI - Make room for computing. PMID- 19815754 TI - Life-long learning for physicians. PMID- 19815755 TI - Conflicting data about dyslexia's cause. PMID- 19815757 TI - Heretical DNA sequences? PMID- 19815758 TI - Comment on "The Arabidopsis circadian clock incorporates a cADPR-based feedback loop". AB - Dodd et al. (Reports, 14 December 2007, p. 1789) reported that the Arabidopsis circadian clock incorporates the signaling molecule cyclic adenosine diphosphate ribose (cADPR). In contrast, we found that there is no rhythm of cADPR levels nor are there any significant effects on the rhythm by cADPR overexpression, thus raising questions about the conclusions of Dodd et al. PMID- 19815759 TI - Megascience. 'Omics data sharing. PMID- 19815761 TI - Neuroscience. Nuclear power for axonal growth. PMID- 19815760 TI - Genomics. Genome project standards in a new era of sequencing. PMID- 19815762 TI - Genetics. Life after GWA studies. PMID- 19815763 TI - Atmospheric science. Monsoons and meltdowns. PMID- 19815764 TI - Development. Aorta's cardinal secret. PMID- 19815765 TI - Paleontology. On the mammalian ear. PMID- 19815766 TI - Physics. Sensing a small but persistent current. PMID- 19815767 TI - Chemistry. Energy flow under control. PMID- 19815768 TI - Reconstituting bacterial RNA repair and modification in vitro. AB - Ribotoxins kill cells by endonucleotically cleaving essential RNAs involved in protein translation. We report here that a stable heterotetramer composed of two bacterial proteins, Pnkp and Hen1, was able to repair transfer RNAs cleaved by ribotoxins in vitro. Before the broken RNAs were ligated by the heterotetramer, a methyl group was added to the 2'-OH group that participated in the original RNA cut. Because of the methylation, RNAs repaired by bacterial Pnkp/Hen1 heterotetramer could not be cleaved again by the ribotoxins. Thus, unlike eukaryotic Hen1 involved in RNA interference, the bacterial Hen1 is part of an RNA repair and modification system. PMID- 19815769 TI - Ice age terminations. AB - 230Th-dated oxygen isotope records of stalagmites from Sanbao Cave, China, characterize Asian Monsoon (AM) precipitation through the ends of the third- and fourthmost recent ice ages. As a result, AM records for the past four glacial terminations can now be precisely correlated with those from ice cores and marine sediments, establishing the timing and sequence of major events. In all four cases, observations are consistent with a classic Northern Hemisphere summer insolation intensity trigger for an initial retreat of northern ice sheets. Meltwater and icebergs entering the North Atlantic alter oceanic and atmospheric circulation and associated fluxes of heat and carbon, causing increases in atmospheric CO2 and Antarctic temperatures that drive the termination in the Southern Hemisphere. Increasing CO2 and summer insolation drive recession of northern ice sheets, with probable positive feedbacks between sea level and CO2. PMID- 19815770 TI - Reactome array: forging a link between metabolome and genome. AB - We describe a sensitive metabolite array for genome sequence-independent functional analysis of metabolic phenotypes and networks, the reactomes, of cell populations and communities. The array includes 1676 dye-linked substrate compounds collectively representing central metabolic pathways of all forms of life. Application of cell extracts to the array leads to specific binding of enzymes to cognate substrates, transformation to products, and concomitant activation of the dye signals. Proof of principle was shown by reconstruction of the metabolic maps of model bacteria. Utility of the array for unsequenced organisms was demonstrated by reconstruction of the global metabolisms of three microbial communities derived from acidic volcanic pool, deep-sea brine lake, and hydrocarbon-polluted seawater. Enzymes of interest are captured on nanoparticles coated with cognate metabolites, sequenced, and their functions unequivocally established. PMID- 19815771 TI - Mapping excited-state dynamics by coherent control of a dendrimer's photoemission efficiency. AB - Adaptive laser pulse shaping has enabled impressive control over photophysical processes in complex molecules. However, the optimal pulse shape that emerges rarely offers straightforward insight into the excited-state properties being manipulated. We have shown that the emission quantum yield of a donor-acceptor macromolecule (a phenylene ethynylene dendrimer tethered to perylene) can be enhanced by 15% through iterative phase modulation of the excitation pulse. Furthermore, by analyzing the pulse optimization process and optimal pulse features, we successfully isolated the dominant elements underlying the control mechanism. We demonstrated that a step function in the spectral phase directs the postexcitation dynamics of the donor moiety, thus characterizing the coherent nature of the donor excited state. An accompanying pump-probe experiment implicates a 2+1 photon control pathway, in which the optimal pulse promotes a delayed excitation to a second excited state through favorable quantum interference. PMID- 19815772 TI - Persistent currents in normal metal rings. AB - Quantum mechanics predicts that the equilibrium state of a resistive metal ring will contain a dissipationless current. This persistent current has been the focus of considerable theoretical and experimental work, but its basic properties remain a topic of controversy. The main experimental challenges in studying persistent currents have been the small signals they produce and their exceptional sensitivity to their environment. We have developed a technique for detecting persistent currents that allows us to measure the persistent current in metal rings over a wide range of temperatures, ring sizes, and magnetic fields. Measurements of both a single ring and arrays of rings agree well with calculations based on a model of non-interacting electrons. PMID- 19815773 TI - The shape and surface variation of 2 Pallas from the Hubble Space Telescope. AB - We obtained Hubble Space Telescope images of 2 Pallas in September 2007 that reveal distinct color and albedo variations across the surface of this large asteroid. Pallas's shape is an ellipsoid with radii of 291 (+/-9), 278 (+/-9), and 250 (+/-9) kilometers, implying a density of 2400 (+/-250) kilograms per cubic meter-a value consistent with a body that formed from water-rich material. Our observations are consistent with the presence of an impact feature, 240 (+/ 25) kilometers in diameter, within Pallas's ultraviolet-dark terrain. Our observations imply that Pallas is an intact protoplanet that has undergone impact excavation and probable internal alteration. PMID- 19815774 TI - Evolutionary development of the middle ear in Mesozoic therian mammals. AB - The definitive mammalian middle ear (DMME) is defined by the loss of embryonic Meckel's cartilage and disconnection of the middle ear from the mandible in adults. It is a major feature distinguishing living mammals from nonmammalian vertebrates. We report a Cretaceous trechnotherian mammal with an ossified Meckel's cartilage in the adult, showing that homoplastic evolution of the DMME occurred in derived therian mammals, besides the known cases of eutriconodonts. The mandible with ossified Meckel's cartilage appears to be paedomorphic. Reabsorption of embryonic Meckel's cartilage to disconnect the ear ossicles from the mandible is patterned by a network of genes and signaling pathways. This fossil suggests that developmental heterochrony and gene patterning are major mechanisms in homplastic evolution of the DMME. PMID- 19815775 TI - Daily electrical silencing in the mammalian circadian clock. AB - Neurons in the brain's suprachiasmatic nuclei (SCNs), which control the timing of daily rhythms, are thought to encode time of day by changing their firing frequency, with high rates during the day and lower rates at night. Some SCN neurons express a key clock gene, period 1 (per1). We found that during the day, neurons containing per1 sustain an electrically excited state and do not fire, whereas non-per1 neurons show the previously reported daily variation in firing activity. Using a combined experimental and theoretical approach, we explain how ionic currents lead to the unusual electrophysiological behaviors of per1 cells, which unlike other mammalian brain cells can survive and function at depolarized states. PMID- 19815776 TI - Comprehensive mapping of long-range interactions reveals folding principles of the human genome. AB - We describe Hi-C, a method that probes the three-dimensional architecture of whole genomes by coupling proximity-based ligation with massively parallel sequencing. We constructed spatial proximity maps of the human genome with Hi-C at a resolution of 1 megabase. These maps confirm the presence of chromosome territories and the spatial proximity of small, gene-rich chromosomes. We identified an additional level of genome organization that is characterized by the spatial segregation of open and closed chromatin to form two genome-wide compartments. At the megabase scale, the chromatin conformation is consistent with a fractal globule, a knot-free, polymer conformation that enables maximally dense packing while preserving the ability to easily fold and unfold any genomic locus. The fractal globule is distinct from the more commonly used globular equilibrium model. Our results demonstrate the power of Hi-C to map the dynamic conformations of whole genomes. PMID- 19815777 TI - Arterial-venous segregation by selective cell sprouting: an alternative mode of blood vessel formation. AB - Blood vessels form de novo (vasculogenesis) or upon sprouting of capillaries from preexisting vessels (angiogenesis). With high-resolution imaging of zebrafish vascular development, we uncovered a third mode of blood vessel formation whereby the first embryonic artery and vein, two unconnected blood vessels, arise from a common precursor vessel. The first embryonic vein formed by selective sprouting of progenitor cells from the precursor vessel, followed by vessel segregation. These processes were regulated by the ligand EphrinB2 and its receptor EphB4, which are expressed in arterial-fated and venous-fated progenitors, respectively, and interact to orient the direction of progenitor migration. Thus, directional control of progenitor migration drives arterial-venous segregation and generation of separate parallel vessels from a single precursor vessel, a process essential for vascular development. PMID- 19815778 TI - KLF family members regulate intrinsic axon regeneration ability. AB - Neurons in the central nervous system (CNS) lose their ability to regenerate early in development, but the underlying mechanisms are unknown. By screening genes developmentally regulated in retinal ganglion cells (RGCs), we identified Kruppel-like factor-4 (KLF4) as a transcriptional repressor of axon growth in RGCs and other CNS neurons. RGCs lacking KLF4 showed increased axon growth both in vitro and after optic nerve injury in vivo. Related KLF family members suppressed or enhanced axon growth to differing extents, and several growth suppressive KLFs were up-regulated postnatally, whereas growth-enhancing KLFs were down-regulated. Thus, coordinated activities of different KLFs regulate the regenerative capacity of CNS neurons. PMID- 19815780 TI - The visual perception of motion by observers with autism spectrum disorders: a review and synthesis. AB - Traditionally, psychological research on autism spectrum disorder (ASD) has focused on social and cognitive abilities. Vision provides an important input channel to both of these processes, and, increasingly, researchers are investigating whether observers with ASD differ from typical observers in their visual percepts. Recently, significant controversies have arisen over whether observers with ASD differ from typical observers in their visual analyses of movement. Initial studies suggested that observers with ASD experience significant deficits in their visual sensitivity to coherent motion in random dot displays but not to point-light displays of human motion. More recent evidence suggests exactly the opposite: that observers with ASD do not differ from typical observers in their visual sensitivity to coherent motion in random dot displays, but do differ from typical observers in their visual sensitivity to human motion. This review examines these apparently conflicting results, notes gaps in previous findings, suggests a potentially unifying hypothesis, and identifies areas ripe for future research. PMID- 19815781 TI - Why does picture-plane inversion sometimes dissociate perception of features and spacing in faces, and sometimes not? Toward a new theory of holistic processing. AB - Classically, it has been presumed that picture-plane inversion primarily reduces sensitivity to spacing/configural information in faces (distance between location of the major features) and has little effect on sensitivity to local feature information (e.g., eye shape or color). Here, we review 22 published studies relevant to this claim. Data show that the feature inversion effect varied substantially across studies as a function of the following factors: whether the feature change was shape only or included color/brightness, the number of faces in the stimulus set, and whether the feature was in facial context. For shape only changes in facial context, feature inversion effects were as large as typical spacing inversion effects. Small feature inversion effects occurred only when a task could be efficiently solved by visual-processing areas outside whole face coding. The results argue that holistic/configural processing for upright faces integrates exact feature shape and spacing between blobs. We describe two plausible approaches to this process. PMID- 19815782 TI - Psychological interpretation of the ex-Gaussian and shifted Wald parameters: a diffusion model analysis. AB - A growing number of researchers use descriptive distributions such as the ex Gaussian and the shifted Wald to summarize response time data for speeded two choice tasks. Some of these researchers also assume that the parameters of these distributions uniquely correspond to specific cognitive processes. We studied the validity of this cognitive interpretation by relating the parameters of the ex Gaussian and shifted Wald distributions to those of the Ratcliff diffusion model, a successful model whose parameters have well-established cognitive interpretations. In a simulation study, we fitted the ex-Gaussian and shifted Wald distributions to data generated from the diffusion model by systematically varying its parameters across a wide range of plausible values. In an empirical study, the two descriptive distributions were fitted to published data that featured manipulations of task difficulty, response caution, and a priori bias. The results clearly demonstrate that the ex-Gaussian and shifted Wald parameters do not correspond uniquely to parameters of the diffusion model. We conclude that researchers should resist the temptation to interpret changes in the ex-Gaussian and shifted Wald parameters in terms of cognitive processes. Supporting materials may be downloaded from http://pbr.psychonomic-journals.org/content/supplemental. PMID- 19815783 TI - Human four-dimensional spatial intuition in virtual reality. AB - It is a long-lasting question whether human beings, who evolved in a physical world of three dimensions, are capable of overcoming this fundamental limitation to develop an intuitive understanding of four-dimensional space. Techniques of analogy and graphical illustration have been developed with some subjective reports of success. However, there has been no objective evaluation of such achievements. Here, we show evidence that people with basic geometric knowledge can learn to make spatial judgments on the length of, and angle between, line segments embedded in four-dimensional space viewed in virtual reality with minimal exposure to the task and no feedback to their responses. Their judgments incorporated information from both the three-dimensional (3-D) projection and the fourth dimension, and the underlying representations were not algebraic in nature but based on visual imagery, although primitive and short lived. These results suggest that human spatial representations are not completely constrained by our evolution and development in a 3-D world. Illustration of the stimuli and experimental procedure (as video clips) and the instruction to participants (as a PDF file) may be downloaded from http://pbr.psychonomic journals.org/content/supplemental. PMID- 19815784 TI - A dissociation between similarity effects in episodic face recognition. AB - Memory similarity, the similarity between a test lure and memory traces, reduces confidence and accuracy in all forms of recognition memory. In contrast, Tulving (1981) showed that, in recognition memory for scenic pictures, choice similarity, the similarity between forced choice test alternatives, increased accuracy but decreased confidence. In the present study, we replicated both memory and choice similarity effects and the dissociation between accuracy and confidence with pictures of faces. State-trace analysis confirmed the dissociation and identified two dimensions underlying these effects, one associated with choice similarity and another associated with memory similarity. Further analysis showed that the effect of study-test lag was associated with the memory-similarity dimension. PMID- 19815785 TI - A stereo disadvantage for recognizing rotated familiar objects. AB - We tested recognition of familiar objects in two different conditions: mono, where stimuli were displayed as flat, 2-D images, and stereo, where objects were displayed with stereoscopic depth information. In three experiments, participants performed a sequential matching task, where an object was rotated by up to 180 degrees between presentations. When the 180 degrees rotation resulted in large changes in depth for object components, recognition performance in the mono condition showed better performance at 180 degrees rotations than at smaller rotations, but stereo presentations showed a monotonic increase in response time with rotation. However, 180 degrees rotations that did not result in much depth variation showed similar patterns of results for mono and stereo conditions. These results suggest that in some circumstances, the lack of explicit 3-D information in 2-D images may influence the recognition of familiar objects when they are depicted on flat computer monitors. PMID- 19815786 TI - Inhibition is picky: shape difference is a necessary condition for attentional inhibition of irrelevant objects. AB - The present study investigated the conditions for attentional amplification in the processing of relevant objects and for attentional inhibition in the processing of irrelevant objects. Participants reported the color of one of two superimposed objects that was specified by occlusion. Irrelevant color words were presented as part of the relevant object, as part of the irrelevant object, or in the background; the words were either congruent or incongruent with the color of the relevant object. The size of the congruency effects provided a measure of the (relative) strength of the processing of the objects and the background. Finally, the two objects had the same shape in one session and different shapes in another session. In both sessions, results showed larger Stroop effects from words belonging to the relevant object than from words in the background, indicating attentional amplification of the relevant object that was unaffected by object similarity. In contrast, smaller Stroop effects from words belonging to the irrelevant object than from words in the background occurred only when the two objects differed in shape. The latter result suggests that a shape difference is both a necessary and a sufficient condition for inhibiting the processing of an irrelevant visual object. PMID- 19815787 TI - Auditory location negative priming: a case of feature mismatch. AB - In an auditory four-alternative forced choice localization task, participants had to localize one of two simultaneously presented sounds while ignoring the location of the second sound. Negative priming--that is, slowed-down responses to a location that had to be ignored in the previous trial--was found only when the sound at the repeated location changed between prime and probe. There was also no increase in prime response errors to the probes of ignored repetition trials. These findings allow for the conclusion that auditory location priming is caused by feature mismatch only and that other mechanisms, such as inhibition of ignored locations or episodic retrieval of transfer-inappropriate prime information, do not play a role. PMID- 19815788 TI - Searching in the dark: cognitive relevance drives attention in real-world scenes. AB - We investigated whether the deployment of attention in scenes is better explained by visual salience or by cognitive relevance. In two experiments, participants searched for target objects in scene photographs. The objects appeared in semantically appropriate locations but were not visually salient within their scenes. Search was fast and efficient, with participants much more likely to look to the targets than to the salient regions. This difference was apparent from the first fixation and held regardless of whether participants were familiar with the visual form of the search targets. In the majority of trials, salient regions were not fixated. The critical effects were observed for all 24 participants across the two experiments. We outline a cognitive relevance framework to account for the control of attention and fixation in scenes. PMID- 19815789 TI - Tracking the train of thought from the laboratory into everyday life: an experience-sampling study of mind wandering across controlled and ecological contexts. AB - In an experience-sampling study that bridged laboratory, ecological, and individual-differences approaches to mind-wandering research, 72 subjects completed an executive-control task with periodic thought probes (reported by McVay & Kane, 2009) and then carried PDAs for a week that signaled them eight times daily to report immediately whether their thoughts were off task. Subjects who reported more mind wandering during the laboratory task endorsed more mind wandering experiences during everyday life (and were more likely to report worries as off-task thought content). We also conceptually replicated laboratory findings that mind wandering predicts task performance: Subjects rated their daily-life performance to be impaired when they reported off-task thoughts, with greatest impairment when subjects' mind wandering lacked metaconsciousness. The propensity to mind wander appears to be a stable cognitive characteristic and seems to predict performance difficulties in daily life, just as it does in the laboratory. PMID- 19815790 TI - The effects of perceptual load on semantic processing under inattention. AB - Inattentional blindness refers to a failure to consciously detect an irrelevant object that appears without any expectation when attention is engaged with another task. The perceptual load theory predicts that task-irrelevant stimuli will reach awareness only when the primary task is of low load, which allows processing resources to spill over to processing task-irrelevant stimuli as well. We studied whether perceptual load has an effect on inattentional blindness for a task-irrelevant stimulus whose meaning is or is not relevant to the attentional goals of the observer. In the critical trial, a word appeared without any expectation in the center of a display of attended pictures. The results showed that, under both high and low load, unexpected words belonging to the attended semantic category were detected more often than semantically unrelated words. These results imply that task-irrelevant stimuli, whose meanings are relevant to the observer's task, enter awareness irrespective of perceptual load. PMID- 19815791 TI - Looking for meaning: eye movements are sensitive to overlapping semantic features, not association. AB - Theories of semantic memory differ in the extent to which relationships among concepts are captured via associative or via semantic relatedness. We examined the contributions of these two factors, using a visual world paradigm in which participants selected the named object from a four-picture display. We controlled for semantic relatedness while manipulating associative strength by using the visual world paradigm's analogue to presenting asymmetrically associated pairs in either their forward or backward associative direction (e.g., ham-eggs vs. eggs ham). Semantically related objects were preferentially fixated regardless of the direction of presentation (and the effect size was unchanged by presentation direction). However, when pairs were associated but not semantically related (e.g., iceberg-lettuce), associated objects were not preferentially fixated in either direction. These findings lend support to theories in which semantic memory is organized according to semantic relatedness (e.g., distributed models) and suggest that association by itself has little effect on this organization. PMID- 19815792 TI - Using puns to study contextual influences on lexical ambiguity resolution: evidence from eye movements. AB - Participants' eye movements were monitored while they read sentences containing biased homographs in either a single-meaning context condition that instantiated the subordinate meaning of the homograph without ruling out the dominant meaning (e.g., "The man with a toothache had a crown made by the best dentist in town") or a dual-meaning pun context condition that supported both the subordinate and dominant meanings (e.g., "The king with a toothache had a crown made by the best dentist in town"). In both of these conditions, the homographs were followed by disambiguating material that supported the subordinate meaning and ruled out the dominant meaning. Fixation times on the homograph were longer in the single meaning condition than in the dual-meaning condition, whereas the reverse pattern was demonstrated for fixation times on the disambiguating region; these effects were observed as early as first-fixation duration. The findings strongly support the reordered access model of lexical ambiguity resolution. PMID- 19815793 TI - Distributional analyses in auditory lexical decision: neighborhood density and word-frequency effects. AB - In the present article, the effects of phonological neighborhood density and word frequency in spoken word recognition were examined using distributional analyses of response latencies in auditory lexical decision. A density x frequency interaction was observed in mean latencies; frequency effects were larger for low density words than for high-density words. Distributional analyses further revealed that for low-density words, frequency effects were reflected in both distributional shifting and skewing, whereas for high-density words, frequency effects were purely mediated by distributional skewing. The results suggest that word frequency plays a role in early auditory word recognition only when there is relatively little competition between similar-sounding words, and that frequency effects in high-density words reflect postlexical checking. PMID- 19815794 TI - What are effective phonological units in Cantonese spoken word planning? AB - Two picture-word interference experiments were conducted to investigate the nature of effective phonological units in Cantonese spoken word production. The names of the pictures were Cantonese monosyllables with a consonant+vowel+consonant (CVC) structure. Participants' picture-naming responses were faster when the target (e.g., "star" /sing1/) and the distractor shared the same CVC component (e.g., /sing4/, meaning "city"), the same CV component (e.g., /sik6/, "eat"), or the same VC component (e.g., /ging2/, "region"), as opposed to when they were unrelated, and the facilitation effects observed were comparable in size. Also, similar facilitation effects were obtained across the CV+tone related and the VC+tone-related conditions, whereas no reliable effect was found in the V+tone-related condition. These results indicate that an effective phonological unit in spoken word planning is neither a syllable (without tone) nor a segmental unit, and that the possible candidates lie between the two, at least in Cantonese. PMID- 19815795 TI - The role of executive function in perspective taking during online language comprehension. AB - During conversation, interlocutors build on the set of shared beliefs known as common ground. Although there is general agreement that interlocutors maintain representations of common ground, there is no consensus regarding whether common ground representations constrain initial language interpretation processes. Here, I propose that executive functioning--specifically, failures in inhibition control--can account for some occasional insensitivities to common-ground information. The present article presents the results of an experiment that demonstrates that individual differences in inhibition control determine the degree to which addressees successfully inhibit perspective-inappropriate interpretations of temporary referential ambiguities in their partner's speech. Whether mentioned information was grounded or not also played a role, suggesting that addressees may show sensitivity to common ground only when it is established collaboratively. The results suggest that, in conversation, perspective information routinely guides online language processing and that occasional insensitivities to perspective can be attributed partly to difficulties in inhibiting perspective-inappropriate interpretations. PMID- 19815796 TI - Does the truth interfere with our ability to deceive? AB - Does the truth interfere with our ability to respond deceptively? We considered this question in the present study by examining the effects of a task set (i.e., selecting truthful or untruthful responses), both by comparing two presentations of the same task and through transfer to a different task. All participants carried out the task either under the instructions to respond both correctly and incorrectly (Experiment 1) or under the instructions to respond truthfully and deceptively (Experiment 2); the order of instructions was counterbalanced. In Experiment 2, after completing the main task, the participants also performed a Stroop task. The findings suggested that deceptive responses took longer overall, regardless of the order of instructions. Moreover, the experience of responding deceptively versus truthfully led to faster responding on a subsequent Stroop test. Although there may well be processes unique to deceptive responding, the evidence suggests that overcoming interference is a process shared by deceptive and nondeceptive tasks. PMID- 19815797 TI - A case for restricted-domain relational learning. AB - Monkeys and pigeons learned a same/different task with pairs that were selected from a training set of eight picture stimuli. They showed no novel-stimulus transfer and hence no abstract-concept learning. They were also tested with novel pairs of the eight training pictures (i.e., combinations that had not been used in training) and with inverted pictures of the training pairs. If the subjects had learned the task item-specifically (e.g., if-then or configural learning), they should have failed these tests, but they performed well with novel combinations of training pictures and inverted pictures, suggesting that they learned the task relationally (i.e., on the basis of the relationship between the two pictures that were presented in each trial). This somewhat paradoxical conclusion of relational learning in the absence of abstract-concept learning is contrary to most theories of abstract-concept learning. The implications of this conclusion are discussed in the context of restricted-domain relational learning. PMID- 19815798 TI - Concepts are not represented by conscious imagery. AB - According to theories of grounded cognition, conceptual representation and perception share processing mechanisms. We investigated whether this overlap is due to conscious perceptual imagery. Participants filled out questionnaires to assess the vividness of their imagery (Questionnaire on Mental Imagery) and the extent to which their imagery was object oriented and spatially oriented (Object Spatial Imagery Questionnaire), and they performed a mental rotation task. One week later, they performed a verbal property verification task. In this task, involvement of modality-specific systems is evidenced by the modality-switch effect, the finding that performance on a target trial (e.g., apple-green) is better after a same-modality trial (e.g., diamond-sparkle) than after a different modality trial (e.g., airplane-noisy). Results showed a modality-switch effect, but there was no systematic relation between imagery scores and modality switch. We conclude that conscious mental imagery is not fundamental to conceptual representation. PMID- 19815799 TI - Five-year-olds' beliefs about the discreteness of category boundaries for animals and artifacts. AB - Research on adult concepts indicates that category structure varies by domain; adults view membership in animal categories as absolute but membership in artifact categories as graded. In this study, we examined domain differences in beliefs about category boundaries among young children (5-year-olds). The results indicated that young children, like adults, were less likely to endorse graded category membership for animal than for artifact categories. These domain differences could not be attributed to domain differences in typicality. Implications for conceptual development and for models of domain specificity in adult cognition are discussed. PMID- 19815800 TI - Children's higher order cognitive abilities and the development of secondary memory. AB - The relations between higher cognitive abilities and immediate and delayed recall were studied in 57 children (6-16 years of age). The participants were tested repeatedly on free recall of a supraspan list (Children's Memory Scale), and their fluid ability was also assessed (Woodcock-Johnson III Spatial Relations). Consistent with Unsworth and Engle's (2007) account of the relation between memory and higher order cognition, the children's fluid ability was significantly correlated with retrieval from secondary memory, regardless of whether it was measured using immediate or delayed recall. Multiple regression analyses provided further support for this view, revealing that measures of immediate and delayed retrieval from secondary memory accounted for the same variance in the children's fluid ability. PMID- 19815801 TI - There's more to the working memory capacity-fluid intelligence relationship than just secondary memory. AB - The present study examined the claim that secondary memory processes account for the correlation between working memory capacity and fluid intelligence via a latent variable analysis. In the present study, participants performed multiple measures of secondary memory, working memory capacity, and fluid intelligence. Structural equation modeling suggested that both secondary memory and working memory capacity account for unique variance in fluid intelligence. These results are inconsistent with recent claims that working memory capacity does not account for variance in fluid intelligence over and above what is accounted for by secondary memory. Rather, the results are consistent with models of working memory capacity that suggest that both maintenance and retrieval processes are needed to account for the substantial relation between working memory capacity and fluid intelligence. PMID- 19815802 TI - Retrieval savings with nonidentical elements: the case of simple addition and subtraction. AB - The identical elements (IE) theory of fact representation (Rickard, 2005) proposes that memorized facts that are composed of identical elements (e.g., 6x8=48 and 8x6=48) share a common representation in memory, whereas facts with nonidentical elements (e.g., 6x8=48 and 48/8=6) are represented separately in memory. The IE model has been successfully applied to the transfer of practice in simple multiplication and division, in transition from procedure-based to retrieval-based performance, and in cued episodic recall. In the present article, we examined the effects of practicing simple addition problems (e.g., 3+6=9) on the performance of corresponding subtraction problems (9-6=3), and vice versa. According to IE theory, there should be no transfer of retrieval savings between addition and subtraction facts if performance is based on discrete IE fact representations. Cross-operation response time savings were observed, however, for both small, well-memorized problems (e.g., practice 3+2, test 5-2) and larger problems (6+8, 14-6), and they were statistically robust when trials that were self-reported as direct retrieval were analyzed. The transfer of retrieval practice savings between facts with nonidentical elements challenges IE theory as a comprehensive model of transfer in memory retrieval. PMID- 19815803 TI - Precise instructions determine participants' memory search strategy in judgments of relative order in short lists. AB - Memory often requires knowledge of the order of events. Previous findings about immediate judgments of relative order in short, subspan lists are variable regarding whether participants' strategy is to search memory in the forward direction, starting from the first list item and progressing toward the end item, or in the backward direction, starting from the end item and progressing toward the start. We asked whether wording of the instructions influences participants' search direction. Participants studied sequences of three to six consonants, and for an immediate, two-item probe of each list, judged which probe was presented earlier ("earlier" instruction) or later ("later" instruction) on the list. Forward and backward searches were supported for "earlier" and "later" instructions, respectively. Our findings suggest that participants have more than one effective strategy for order judgments in short lists, and that subtle instructional differences can bias memory search in either the forward or backward direction. PMID- 19815804 TI - Revelation effect in metamemory. AB - Test items are more likely to be judged as previously studied if they need to be discovered before the recognition decision. In the present experiments, this revelation effect was extended to metamemory judgments. Participants studied word pairs and then tried to recall the second word of each pair when given the first word as a cue. In Experiment 1, a fragment of the target was either gradually increased in size or held constant, and in Experiment 2, sometimes an anagram of the cue was given instead of the cue itself. Thus, for some items, there was a revelation task before a recall attempt. If recall failed, the participants gave feeling-of-knowing (FOK) ratings. In both experiments, the participants gave higher FOK ratings after a revelation task, even though the items that these FOKs referred to remained unrecalled. Analyses showed a criterion shift but no differences in sensitivity. PMID- 19815805 TI - Navigating through abstract decision spaces: evaluating the role of state generalization in a dynamic decision-making task. AB - Research on dynamic decision-making tasks, in which the payoffs associated with each choice vary with participants' recent choice history, shows that humans have difficulty making long-term optimal choices in the presence of attractive immediate rewards. However, a number of recent studies have shown that simple cues providing information about the underlying state of the task environment may facilitate optimal responding. In this study, we examined the mechanism by which this state knowledge influences choice behavior. We examined the possibility that participants use state information in conjunction with changing payoffs to extrapolate payoffs in future states. We found support for this hypothesis in an experiment in which generalizations based on this state information worked to the benefit or detriment of task performance, depending on the task's payoff structure. PMID- 19815806 TI - Who is being deceived? The experimental demands of wearing a backpack. AB - A growing literature argues that wearing a heavy backpack makes slopes look steeper and distances seem longer (e.g., Proffitt, 2006). To test for effects of experimental demand characteristics in a backpack experiment, we manipulated the experimental demand of the backpack and then used a postexperiment questionnaire to assess participants' beliefs about the purpose of the backpack. For participants in the low-demand condition, an elaborate deception was used to provide an alternative explanation of the requirement to wear a heavy backpack (i.e., that it held EMG equipment). The highest slope judgments were found for those undeceived participants who guessed that the backpack was intended to affect their slope perception and also reported that they thought they were affected by it. When persuaded that the backpack served another purpose, participants' slope estimates were no different from those of participants not wearing a backpack. These findings suggest that backpack effects, and other reported effects of effort on perception, are judgmental biases that result from the social, not physical, demands of the experimental context. PMID- 19815807 TI - Affordances matter in geographical slant perception. AB - We argue that the experimental conditions in the Durgin et al. (2009) study were so different from those in Bhalla and Proffitt (1999) that the results of the former study cannot be generalized to the latter. The participants in the Durgin et al. study viewed a 2-m-long ramp; those in Bhalla and Proffitt viewed expansive hills. When drawing generalizations from one study to another, equating experimental conditions is always important; moreover, from an embodied perspective on perception, equating the opportunities for action also matters. PMID- 19815808 TI - Transcriptomic analysis of human lung development. AB - RATIONALE: Current understanding of the molecular regulation of lung development is limited and derives mostly from animal studies. OBJECTIVES: To define global patterns of gene expression during human lung development. METHODS: Genome-wide expression profiling was used to measure the developing lung transcriptome in RNA samples derived from 38 normal human lung tissues at 53 to 154 days post conception. Principal component analysis was used to characterize global expression variation and to identify genes and bioontologic attributes contributing to these variations. Individual gene expression patterns were verified by quantitative reverse transcriptase-polymerase chain reaction analysis. MEASUREMENTS AND MAIN RESULTS: Gene expression analysis identified attributes not previously associated with lung development, such as chemokine immunologic processes. Lung characteristics attributes (e.g., surfactant function) were observed at an earlier-than-anticipated age. We defined a 3,223 gene developing lung characteristic subtranscriptome capable of describing a majority of the process. In gene expression space, the samples formed a time contiguous trajectory with transition points correlating with histological stages and suggesting the existence of novel molecular substages. Induction of surfactant gene expression characterized a pseudoglandular "molecular phase" transition. Individual gene expression patterns were independently validated. We predicted the age of independent human lung transcriptome profiles with a median absolute error of 5 days, supporting the validity of the data and modeling approach. CONCLUSIONS: This study extends our knowledge of key gene expression patterns and bioontologic attributes underlying early human lung developmental processes. The data also suggest the existence of molecular phases of lung development. PMID- 19815809 TI - Effectiveness and safety of bronchial thermoplasty in the treatment of severe asthma: a multicenter, randomized, double-blind, sham-controlled clinical trial. AB - RATIONALE: Bronchial thermoplasty (BT) is a bronchoscopic procedure in which controlled thermal energy is applied to the airway wall to decrease smooth muscle. OBJECTIVES: To evaluate the effectiveness and safety of BT versus a sham procedure in subjects with severe asthma who remain symptomatic despite treatment with high-dose inhaled corticosteroids and long-acting beta(2)-agonists. METHODS: A total of 288 adult subjects (Intent-to-Treat [ITT]) randomized to BT or sham control underwent three bronchoscopy procedures. Primary outcome was the difference in Asthma Quality of Life Questionnaire (AQLQ) scores from baseline to average of 6, 9, and 12 months (integrated AQLQ). Adverse events and health care use were collected to assess safety. Statistical design and analysis of the primary endpoint was Bayesian. Target posterior probability of superiority (PPS) of BT over sham was 95%, except for the primary endpoint (96.4%). MEASUREMENTS AND MAIN RESULTS: The improvement from baseline in the integrated AQLQ score was superior in the BT group compared with sham (BT, 1.35 +/- 1.10; sham, 1.16 +/- 1.23 [PPS, 96.0% ITT and 97.9% per protocol]). Seventy-nine percent of BT and 64% of sham subjects achieved changes in AQLQ of 0.5 or greater (PPS, 99.6%). Six percent more BT subjects were hospitalized in the treatment period (up to 6 wk after BT). In the posttreatment period (6-52 wk after BT), the BT group experienced fewer severe exacerbations, emergency department (ED) visits, and days missed from work/school compared with the sham group (PPS, 95.5, 99.9, and 99.3%, respectively). CONCLUSIONS: BT in subjects with severe asthma improves asthma-specific quality of life with a reduction in severe exacerbations and healthcare use in the posttreatment period. Clinical trial registered with www.clinialtrials.gov (NCT00231114). PMID- 19815810 TI - Specific CD8 T cells in IgE-mediated allergy correlate with allergen dose and allergic phenotype. AB - RATIONALE: Studies in humans and rodents have indicated a causative role for CD8(+) T cells in IgE-mediated allergic inflammation, but their function is still controversial. OBJECTIVES: To analyze the role of allergen-specific CD8(+) T cells during the development of allergic airway inflammation in two parallel but diverging outcome models. METHODS: We used H2-Kb SIINFEKL (OVA(257-264)) multimers to analyze induction, natural distribution, and phenotype of allergen specific CD8(+) T cells in a murine C57BL/6 model of ovalbumin (OVA)-induced allergic airway inflammation using low-dose or high-dose OVA sensitization. MEASUREMENTS AND MAIN RESULTS: The low-dose protocol was characterized by a significant induction of total and OVA-specific IgE, eosinophilic airway inflammation, IL-4 levels in bronchoalveolar lavage fluid. And significant alterations in lung function. The high dose protocol was characterized by a significant reduction of the allergic phenotype. Using OVA(257-264) H2-Kb multimers, we observed lung and airway infiltrating OVA-specific CD8(+) T cells showing an effector/effector-memory phenotype. The high-dose protocol caused significantly higher infiltration of allergen-specific CD8(+) cells to the airways and enhanced their cytotoxicity. Adoptive transfer with CD8(+) T cells from transgenic OT-I mice to TAP1(-/-) or wild-type mice showed their migration to the lungs and TAP1-dependent proliferation after OVA-aerosol exposure. TAP1(-/ ) mice defective in CD8(+) T cells showed exacerbated symptoms in the low-dose sensitization model. CONCLUSIONS: Allergen-specific CD8(+) T cells seem to protect from allergic inflammation in the lungs. Their number, which is dependent on the sensitization dose, appears to be a critical predictor for the severity of the allergic phenotype. PMID- 19815811 TI - Regional differential effects of the novel histamine H3 receptor antagonist 6-[(3 cyclobutyl-2,3,4,5-tetrahydro-1H-3-benzazepin-7-yl)oxy]-N-methyl-3 pyridinecarboxamide hydrochloride (GSK189254) on histamine release in the central nervous system of freely moving rats. AB - After oral administration, the nonimidazole histamine H(3) receptor antagonist, 6 [(3-cyclobutyl-2,3,4,5-tetrahydro-1H-3-benzazepin-7-yl)oxy]-N-methyl-3 pyridinecarboxamide hydrochloride (GSK189254), increased histamine release from the tuberomammillary nucleus, where all histaminergic somata are localized, and from where their axons project to the entire brain. To further understand functional histaminergic circuitry in the brain, dual-probe microdialysis was used to pharmacologically block H(3) receptors in the tuberomammillary nucleus, and monitor histamine release in projection areas. Perfusion of the tuberomammillary nucleus with GSK189254 increased histamine release from the tuberomammillary nucleus, nucleus basalis magnocellularis, and cortex, but not from the striatum or nucleus accumbens. Cortical acetylcholine (ACh) release was also increased, but striatal dopamine release was not affected. When administered locally, GSK189254 increased histamine release from the nucleus basalis magnocellularis, but not from the striatum. Thus, defined by their sensitivity to GSK189254, histaminergic neurons establish distinct pathways according to their terminal projections, and can differentially modulate neurotransmitter release in a brain region-specific manner. Consistent with its effects on cortical ACh release, systemic administration of GSK189254 antagonized the amnesic effects of scopolamine in the rat object recognition test, a cognition paradigm with important cortical components. PMID- 19815812 TI - NF546 [4,4'-(carbonylbis(imino-3,1-phenylene-carbonylimino-3,1-(4-methyl phenylene)-carbonylimino))-bis(1,3-xylene-alpha,alpha'-diphosphonic acid) tetrasodium salt] is a non-nucleotide P2Y11 agonist and stimulates release of interleukin-8 from human monocyte-derived dendritic cells. AB - The G protein-coupled P2Y(11) receptor is involved in immune system modulation. In-depth physiological evaluation is hampered, however, by a lack of selective and potent ligands. By screening a library of sulfonic and phosphonic acid derivatives at P2Y(11) receptors recombinantly expressed in human 1321N1 astrocytoma cells (calcium and cAMP assays), the selective non-nucleotide P2Y(11) agonist NF546 [4,4'-(carbonylbis(imino-3,1-phenylene-carbonylimino-3,1-(4-methyl phenylene)carbonylimino))-bis(1,3-xylene-alpha,alpha'-diphosphonic acid) tetrasodium salt] was identified. NF546 had a pEC(50) of 6.27 and is relatively selective for P2Y(11) over P2Y(1), P2Y(2), P2Y(4), P2Y(6), P2Y(12), P2X(1), P2X(2), and P2X(2)-X(3). Adenosine-5'-O-(3-thio)triphosphate (ATPgammaS), a nonhydrolyzable analog of the physiological P2Y(11) agonist ATP, and NF546 use a common binding site as suggested by molecular modeling studies and their competitive behavior toward the nanomolar potency antagonist NF340 [4,4' (carbonylbis(imino-3,1-(4-methyl-phenylene)carbonylimino))bis(naphthalene-2,6 disulfonic acid) tetrasodium salt] in Schild analysis. The pA(2) of NF340 was 8.02 against ATPgammaS and 8.04 against NF546 (calcium assays). NF546 was further tested for P2Y(11)-mediated effects in monocyte-derived dendritic cells. Similarly to ATPgammaS, NF546 led to thrombospondin-1 secretion and inhibition of lipopolysaccharide-stimulated interleukin-12 release, whereas NF340 inhibited these effects. Further, for the first time, it was shown that ATPgammaS or NF546 stimulation promotes interleukin 8 (IL-8) release from dendritic cells, which could be inhibited by NF340. In conclusion, we have described the first selective, non-nucleotide agonist NF546 for P2Y(11) receptors in both recombinant and physiological expression systems and could show a P2Y(11)-stimulated IL-8 release, further supporting the immunomodulatory role of P2Y(11) receptors. PMID- 19815813 TI - Early intervention of tyrosine nitration prevents vaso-obliteration and neovascularization in ischemic retinopathy. AB - Diabetic retinopathy and retinopathy of prematurity are blinding disorders that follow a pathological pattern of ischemic retinopathy and affect premature infants and working-age adults. Yet, the treatment options are limited to laser photocoagulation. The goal of this study is to elucidate the molecular mechanism and examine the therapeutic effects of inhibiting tyrosine nitration on protecting early retinal vascular cell death and late neovascularization in the ischemic retinopathy model. Ischemic retinopathy was developed by exposing neonatal mice to 75% oxygen [postnatal day (p) 7-p12] followed by normoxia (21% oxygen) (p12-p17). Peroxynitrite decomposition catalyst 5,10,15,20-tetrakis(4 sulfonatophenyl)porphyrinato iron III chloride (FeTPPS) (1 mg/kg), the nitration inhibitor epicatechin (10 mg/kg) or the thiol donor N-acetylcysteine (NAC, 150 mg/kg) were administered (p7-p12) or (p7-p17). Vascular endothelial cells were incubated at hyperoxia (40% oxygen) or normoxia (21% oxygen) for 48 h. Vascular density was determined in retinal flat mounts labeled with isolectin B4. Expression of vascular endothelial growth factor, caspase-3, and poly(ADP ribose) polymerase (PARP), activation of Akt and p38 mitogen-activated protein kinase (MAPK), and tyrosine nitration of the phosphatidylinositol (PI) 3-kinase p85 subunit were analyzed by Western blot. Hyperoxia-induced peroxynitrite caused endothelial cell apoptosis as indicated by expression of cleaved caspase-3 and PARP leading to vaso-obliteration. These effects were associated with significant tyrosine nitration of the p85 subunit of PI 3-kinase, decreased Akt activation, and enhanced p38 MAPK activation. Blocking tyrosine nitration of PI 3-kinase with epicatechin or NAC restored Akt phosphorylation, and inhibited vaso-obliteration at p12 and neovascularization at p17 comparable with FeTPPS. Early inhibition of tyrosine nitration with use of epicatechin or NAC can represent safe and effective vascular-protective agents in ischemic retinopathy. PMID- 19815814 TI - Rare autosomal dominant POLG1 mutation in a family with metabolic strokes, posterior column spinal degeneration, and multi-endocrine disease. AB - DNA POLG is the only mitochondrial DNA polymerase and is encoded by nuclear DNA. Depending on the location and inheritance, mutations in POLG1, the catalytic subunit, can cause symptoms including severe infantile epilepsy, metabolic strokes, chronic ataxia, neuropathy, and ophthalmoplegia. We reviewed medical records and conducted extensive interviews with the family of identical twin probands with a mutation in the linker region of DNA polymerase gamma 1 (POLG1) (G517V) and discuss postmortem findings from their grandmother. Both twins developed type I diabetes, adrenal insufficiency, hypothyroidism, and psychiatric problems in addition to neurological difficulties including bilateral basal ganglia infarcts, headaches, and seizures. The maternal grandmother, now deceased, had psychosis and balance problems, and postmortem findings include lacunar infarcts in the basal ganglia (caudate nucleus, putamen, and globus pallidus) and posterior spinal column degeneration. We discuss novel aspects of their presentation and implications for practice. PMID- 19815815 TI - Chylomicronemia elicits atherosclerosis in mice--brief report. AB - OBJECTIVE: The risk of atherosclerosis in the setting of chylomicronemia has been a topic of debate. In this study, we examined susceptibility to atherosclerosis in Gpihbp1-deficient mice (Gpihbp1(-/-)), which manifest severe chylomicronemia as a result of defective lipolysis. METHODS AND RESULTS: Gpihbp1(-/-) mice on a chow diet have plasma triglyceride and cholesterol levels of 2812+/-209 and 319+/ 27 mg/dL, respectively. Even though nearly all of the lipids were contained in large lipoproteins (50 to 135 nm), the mice developed progressive aortic atherosclerosis. In other experiments, we found that both Gpihbp1-deficient "apo B48-only" mice and Gpihbp1-deficient "apo-B100-only" mice manifest severe chylomicronemia. Thus, GPIHBP1 is required for the processing of both apo-B48- and apo-B100-containing lipoproteins. CONCLUSIONS: Chylomicronemia causes atherosclerosis in mice. Also, we found that GPIHBP1 is required for the lipolytic processing of both apo-B48- and apo-B100-containing lipoproteins. PMID- 19815816 TI - Effects of cigarette smoke exposure on clot dynamics and fibrin structure: an ex vivo investigation. AB - OBJECTIVE: The purpose of this study was to examine the effect of cigarette smoke exposure (CSE) on clot dynamics and fibrin architecture and to isolate the relative contribution of platelets and fibrinogen to clot dynamics. METHODS AND RESULTS: From young healthy males smokers (n=34) and nonsmokers (n=34) a baseline blood was drawn, and smokers had another blood draw after smoking 2 regular cigarettes. Using thromboelastography (TEG) the degree of platelet-fibrin interaction was measured. In additional experiments, abciximab (20 microg/mL) was added to the smokers samples (n=27) to reduce the effects of platelet function from the TEG parameters. The maximum clot strength (G) obtained with abciximab measured mainly the contribution of fibrinogen to clot strength (GF). By subtracting GF from G, the contribution of platelets to clot strength (GP) was presumed. A significant difference was found for all TEG parameters between nonsmokers versus postsmoking and pre- versus postsmoking samples. Postsmoking both GF and GP were significantly higher as compared to presmoking. On electron microscopy and turbidity analysis, postsmoking fibrin clots were significantly different compared to presmoking and nonsmoking samples. CONCLUSIONS: Acute CSE changes clot dynamics and alters fibrin architecture. Both functional changes in fibrinogen and platelets appear to contribute to heightened thrombogenicity after acute CSE. PMID- 19815818 TI - Alzheimer disease-associated peptide, amyloid beta40, inhibits vascular regeneration with induction of endothelial autophagy. AB - OBJECTIVE: Although the majority of cases of Alzheimer disease (AD) are known to be attributable to the sporadic (nongenetic) form of the disease, the mechanism underlying its cause and progression still remains unclear. METHODS AND RESULTS: We found that vascular beta-amyloid (Abeta), Abeta40, inhibited the proliferative activity of human brain vascular endothelial cells (HBECs) without toxic effects on them. This peptide also inhibited tube formation and migration of HBECs. Moreover, Abeta40 inhibited ex vivo hippocampal revascularization, reendothelialization, and the differentiation of adult endothelial progenitor cells. Importantly, Abeta40 suppressed the proliferative activity of HBECs through the induction of "self-digesting" autophagy. This induction involved the intracellular regulation of class 3 phosphatidylinositol 3-kinase (PI3K) as well as Akt signaling in HBECs. Furthermore, tissue culture of murine brain sections from GFP-LC3 transgenic mice revealed that Abeta40 not only reduced the vessel density in hippocampal lesions, but also induced autophagy in neurovascular ECs. CONCLUSIONS: Our present findings indicate that the initial progression of AD might be in part driven by Abeta40-induced endothelial autophagy and impairment of neurovascular regeneration, suggesting important implications for therapeutic approaches to AD. PMID- 19815817 TI - Increased HDL cholesterol and apoA-I in humans and mice treated with a novel SR BI inhibitor. AB - OBJECTIVE: Increasing HDL levels is a potential strategy for the treatment of atherosclerosis. METHODS AND RESULTS: ITX5061, a molecule initially characterized as a p38 MAPK inhibitor, increased HDL-C levels by 20% in a human population of hypertriglyceridemic subjects with low HDL levels. ITX5061 also moderately increased apoA-I but did not affect VLDL/LDL cholesterol or plasma triglyceride concentrations. ITX5061 increased HDL-C in WT and human apoA-I transgenic mice, and kinetic experiments showed that ITX5061 decreased the fractional catabolic rate of HDL-CE and reduced its hepatic uptake. In transfected cells, ITX5061 inhibited SR-BI-dependent uptake of HDL-CE. Moreover, ITX5061 failed to increase HDL-C levels in SR-BI(-/-) mice. To assess effects on atherosclerosis, ITX5061 was given to atherogenic diet-fed Ldlr(+/-) mice with or without CETP expression for 18 weeks. In both the control and CETP-expressing groups, ITX5061-treated mice displayed reductions of early atherosclerotic lesions in the aortic arch 40%, P<0.05), and a nonsignificant trend to reduced lesion area in the proximal aorta. CONCLUSIONS: Our data indicate that ITX5061 increases HDL-C levels by inhibition of SR-BI activity. This suggests that pharmacological inhibition of SR BI has the potential to raise HDL-C and apoA-I levels without adverse effects on VLDL/LDL cholesterol levels in humans. PMID- 19815819 TI - High-density lipoprotein-based contrast agents for multimodal imaging of atherosclerosis. AB - Lipoproteins, natural nanoparticles, have a well-recognized biological role and are highly suitable as a platform for delivering imaging agents. The ease with which both the exterior and interior of the particles can be modified permits the creation of multifunctional nanoparticles for imaging as well as the delivery of therapeutics. Importantly, their endogenous nature may make them biocompatible and biodegradable and allows them to avoid the recognition of the reticuloendothelial system. In particular, high-density lipoproteins (HDL) are of interest, because of their small size they can easily cross the endothelium and penetrate the underlying tissue. We summarize here the progress in establishing HDL as a vector for delivering a variety of diagnostically active materials to vulnerable atherosclerotic plaques in mouse models of atherosclerosis. By loading various types of image-enhancing compounds into either the core or surface of HDL, they can be visualized by different imaging modalities (MRI, CT, optical). By rerouting of HDL away from plaque macrophages, imaging of biological processes in diseases besides atherosclerosis may also be achieved. PMID- 19815820 TI - Progenitor cells from the explanted heart generate immunocompatible myocardium within the transplanted donor heart. AB - RATIONALE: Chronic rejection, accelerated coronary atherosclerosis, myocardial infarction, and ischemic heart failure determine the unfavorable evolution of the transplanted heart in humans. OBJECTIVE: Here we tested whether the pathological manifestations of the transplanted heart can be corrected partly by a strategy that implements the use of cardiac progenitor cells from the recipient to repopulate the donor heart with immunocompatible cardiomyocytes and coronary vessels. METHODS AND RESULTS: A large number of cardiomyocytes and coronary vessels were created in a rather short period of time from the delivery, engraftment, and differentiation of cardiac progenitor cells from the recipient. A proportion of newly formed cardiomyocytes acquired adult characteristics and was integrated structurally and functionally within the transplant. Similarly, the regenerated arteries, arterioles, and capillaries were operative and contributed to the oxygenation of the chimeric myocardium. Attenuation in the extent of acute damage by repopulating cardiomyocytes and vessels decreased significantly the magnitude of myocardial scarring preserving partly the integrity of the donor heart. CONCLUSIONS: Our data suggest that tissue regeneration by differentiation of recipient cardiac progenitor cells restored a significant portion of the rejected donor myocardium. Ultimately, immunosuppressive therapy may be only partially required improving quality of life and lifespan of patients with cardiac transplantation. PMID- 19815821 TI - Metabolic remodeling induced by mitochondrial aldehyde stress stimulates tolerance to oxidative stress in the heart. AB - RATIONALE: Aldehyde accumulation is regarded as a pathognomonic feature of oxidative stress-associated cardiovascular disease. OBJECTIVE: We investigated how the heart compensates for the accelerated accumulation of aldehydes. METHODS AND RESULTS: Aldehyde dehydrogenase 2 (ALDH2) has a major role in aldehyde detoxification in the mitochondria, a major source of aldehydes. Transgenic (Tg) mice carrying an Aldh2 gene with a single nucleotide polymorphism (Aldh2*2) were developed. This polymorphism has a dominant-negative effect and the Tg mice exhibited impaired ALDH activity against a broad range of aldehydes. Despite a shift toward the oxidative state in mitochondrial matrices, Aldh2*2 Tg hearts displayed normal left ventricular function by echocardiography and, because of metabolic remodeling, an unexpected tolerance to oxidative stress induced by ischemia/reperfusion injury. Mitochondrial aldehyde stress stimulated eukaryotic translation initiation factor 2alpha phosphorylation. Subsequent translational and transcriptional activation of activating transcription factor-4 promoted the expression of enzymes involved in amino acid biosynthesis and transport, ultimately providing precursor amino acids for glutathione biosynthesis. Intracellular glutathione levels were increased 1.37-fold in Aldh2*2 Tg hearts compared with wild-type controls. Heterozygous knockout of Atf4 blunted the increase in intracellular glutathione levels in Aldh2*2 Tg hearts, thereby attenuating the oxidative stress-resistant phenotype. Furthermore, glycolysis and NADPH generation via the pentose phosphate pathway were activated in Aldh2*2 Tg hearts. (NADPH is required for the recycling of oxidized glutathione.) CONCLUSIONS: The findings of the present study indicate that mitochondrial aldehyde stress in the heart induces metabolic remodeling, leading to activation of the glutathione-redox cycle, which confers resistance against acute oxidative stress induced by ischemia/reperfusion. PMID- 19815822 TI - ASK1 regulates cardiomyocyte death but not hypertrophy in transgenic mice. AB - RATIONALE: Apoptosis signal-regulating kinase (ASK)1 is a central upstream kinase in the greater mitogen-activated protein kinase cascade that mediates growth and death decisions in cardiac myocytes in response to diverse pathological stimuli. OBJECTIVE: However, the role that ASK1 plays in regulating the cardiac hypertrophic response in vivo remains controversial. METHODS AND RESULTS: Here, we generated mice with cardiac-specific and inducible overexpression of ASK1 in the heart to assess its gain-of-function effect. ASK1 transgenic mice exhibited no induction of cardiac hypertrophy or pathology at 3 and 12 months of age, and these mice showed an identical hypertrophic response to controls following 2 weeks of pressure-overload stimulation or isoproterenol infusion. Although ASK1 overexpression did not alter the cardiac hypertrophic response, it promoted cardiomyopathy and greater TUNEL following pressure-overload stimulation and myocardial infarction. Indeed, ASK1 transgenic mice showed a greater than 2-fold increase in ischemia reperfusion-induced injury to the heart compared with controls. Examination of downstream signaling showed a prominent activation of mitogen-activated protein kinase kinase 4/6 and c-Jun NH(2)-terminal kinase (JNK)1/2 (but not p38 or extracellular signal-regulated kinases [ERKs]), inhibition of calcineurin-NFAT (nuclear factor of activated T cells), and induction of Bax in the hearts of ASK1 transgenic mice following 1 and 8 weeks of pressure-overload stimulation. Mechanistically, cardiomyopathy associated with ASK1 overexpression after 8 weeks of pressure overload was significantly reduced in the calcineurin Abeta-null (CnAbeta(-/-)) background. CONCLUSIONS: These results indicate that ASK1 does not directly regulate the cardiac hypertrophic response in vivo, but it does alter cell death and propensity to cardiomyopathy, in part, through a calcineurin-dependent mechanism. PMID- 19815823 TI - Conditional ablation of nonmuscle myosin II-B delineates heart defects in adult mice. AB - RATIONALE: Germline ablation of the cytoskeletal protein nonmuscle myosin II (NMII)-B results in embryonic lethality, with defects in both the brain and heart. Tissue-specific ablation of NMII-B by a Cre recombinase strategy should prevent embryonic lethality and permit study of the function of NMII-B in adult hearts. OBJECTIVE: We sought to understand the function of NMII-B in adult mouse hearts and to see whether the brain defects found in germline-ablated mice influence cardiac development. METHODS AND RESULTS: We used a loxP/Cre recombinase strategy to specifically ablate NMII-B in the brains or hearts of mice. Mice ablated for NMII-B in neural tissues die between postnatal day 12 and 22 without showing cardiac defects. Mice deficient in NMII-B only in cardiac myocytes (B(alphaMHC)/B(alphaMHC) mice) do not show brain defects. However, B(alphaMHC)/B(alphaMHC) mice display novel cardiac defects not seen in NMII-B germline-ablated mice. Most of the B(alphaMHC)/B(alphaMHC) mice are born with enlarged cardiac myocytes, some of which are multinucleated, reflecting a defect in cytokinesis. Between 6 to 10 months, they develop a cardiomyopathy that includes interstitial fibrosis and infiltration of the myocardium and pericardium with inflammatory cells. Four of 5 B(alphaMHC)/B(alphaMHC) hearts develop marked widening of intercalated discs. CONCLUSIONS: By avoiding the embryonic lethality found in germline-ablated mice, we were able to study the function of NMII-B in adult mice and show that absence of NMII-B in cardiac myocytes results in cardiomyopathy in the adult heart. We also define a role for NMII-B in maintaining the integrity of intercalated discs. PMID- 19815824 TI - Complement-dependent inflammation and injury in a murine model of brain dead donor hearts. AB - RATIONALE: Donor brain death (BD) is an unavoidable occurrence in heart transplantation and results in profound physiological derangements that render the heart more susceptible to ischemia/reperfusion injury in the recipient and likely has negative long-term consequences to allograft survival. OBJECTIVE: We developed a novel mouse model of BD and investigated the role of complement in BD induced myocardial inflammation and injury. METHODS AND RESULTS: BD was induced by inflation of a balloon catheter in the cranial cavity. BD in wild-type mice resulted in a significant increase in serum concentrations of the complement activation product complement component (C)3a, and immunohistochemical analysis of heart sections demonstrated C3 deposition on the vascular endothelium and surrounding myocytes. Following induction of BD in complement (C3)-deficient mice, cardiac troponin levels, and histological evidence of injury were significantly reduced compared to wild-type mice. C3 deficiency was also associated with reduced myocardial leukocyte infiltration and reduced or absent expression of P-selectin, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, tumor necrosis factor-alpha, and interleukin-1beta. CONCLUSIONS: These data indicate an important role for complement in BD-induced inflammation and injury and suggest that a complement inhibitory strategy applied to the donor (in addition to the recipient) may provide graft protection. PMID- 19815825 TI - Spiral waves and reentry dynamics in an in vitro model of the healed infarct border zone. AB - RATIONALE: Reentry underlies most ventricular tachycardias (VTs) seen postmyocardial infarction (MI). Mapping studies reveal that the majority of VTs late post-MI arise from the infarct border zone (IBZ). OBJECTIVE: To investigate reentry dynamics and the role of individual ion channels on reentry in in vitro models of the "healed" IBZ. METHODS AND RESULTS: We designed in vitro models of the healed IBZ by coculturing skeletal myotubes with neonatal rat ventricular myocytes and performed optical mapping at high temporal and spatial resolution. In culture, neonatal rat ventricular myocytes mature to form striated myocytes and electrically uncoupled skeletal myotubes simulate fibrosis seen in the healed IBZ. High resolution mapping revealed that skeletal myotubes produced localized slowing of conduction velocity (CV), increased dispersion of CV and directional dependence of activation delay without affecting myocyte excitability. Reentry was easily induced by rapid pacing in cocultures; treatment with lidocaine, a Na(+) channel blocker, significantly decreased reentry rate and CV, increased reentry path length and terminated 30% of reentrant arrhythmias (n=18). In contrast, nitrendipine, an L-type Ca(2+) channel blocker terminated 100% of reentry episodes while increasing reentry cycle length and path length and decreasing reentry CV (n=16). K(+) channel blockers increased reentry action potential duration but infrequently terminated reentry (n=12). CONCLUSIONS: Cocultures reproduce several architectural and electrophysiological features of the healed IBZ. Reentry termination by L-type Ca(2+) channel, but not Na(+) channel, blockers suggests a greater Ca(2+)-dependence of propagation. These results may help explain the low efficacy of pure Na(+) channel blockers in preventing and terminating clinical VTs late after MI. PMID- 19815826 TI - Cardiac metabolic state and Brugada syndrome: a link revealed. PMID- 19815828 TI - Increased risk of stroke after a herpes zoster attack: a population-based follow up study. AB - BACKGROUND AND PURPOSE: Varicella zoster virus-induced vasculopathy and postherpes zoster attack stroke syndromes have been reported previously; nevertheless, data regarding the exact prevalence and risk of stroke occurring postherpes zoster attack are still lacking. This study aims to investigate the frequency and risk of stroke after a herpes zoster attack using a nationwide, population-based study of a retrospective cohort design. Method- A total of 7760 patients who had received treatment for herpes zoster between 1997 and 2001 were included and matched with 23 280 randomly selected subjects. A 1-year stroke-free survival rate was then estimated using the Kaplan-Meier method. After adjusting for potential confounders, Cox proportional hazard regressions were carried out to compute the adjusted 1-year survival rate. RESULTS: Of the sampled patients, 439 patients (1.41%) developed strokes within the 1-year follow-up period, that is, 133 individuals (1.71% of the patients with herpes zoster) from the study cohort and 306 individuals (1.31% of patients in the comparison cohort) from the comparison cohort. The log rank test indicated that patients with herpes zoster had significantly lower 1-year stroke-free survival rates than the control (P<0.001). The adjusted hazard ratios of stroke after herpes zoster and herpes zoster ophthalmicus during the 1-year follow-up period were 1.31 and 4.28, respectively. CONCLUSIONS: The risk for stroke increased after a zoster attack. Although varicella zoster virus vasculopathy is a well-documented complication that may induce a stroke postherpes zoster attack, it does not fully account for the unexpectedly high risk of stroke in these patients. PMID- 19815827 TI - Human studies of angiogenic gene therapy. AB - Despite significant advances in medical, interventional, and surgical therapy for coronary and peripheral arterial disease, the burden of these illnesses remains high. To address this unmet need, the science of therapeutic angiogenesis has been evolving for almost two decades. Early preclinical studies and phase I clinical trials achieved promising results with growth factors administered as recombinant proteins or as single-agent gene therapies, and data accumulated through 10 years of clinical trials indicate that gene therapy has an acceptable safety profile. However, more rigorous phase II and phase III clinical trials have failed to unequivocally demonstrate that angiogenic agents are beneficial under the conditions and in the patients studied to date. Investigators have worked to understand the biology of the vascular system and to incorporate their findings into new treatments for patients with ischemic disease. Recent gene- and cell-therapy trials have demonstrated the bioactivity of several new agents and treatment strategies. Collectively, these observations have renewed interest in the mechanisms of angiogenesis and deepened our understanding of the complexity of vascular regeneration. Gene therapy that incorporates multiple growth factors, approaches that combine cell and gene therapy, and the administration of "master switch" agents that activate numerous downstream pathways are among the credible and plausible steps forward. In this review, we examine the clinical development of angiogenic gene therapy, summarize several of the lessons learned during the conduct of these trials, and suggest how this prior experience may guide the conduct of future preclinical investigations and clinical trials. PMID- 19815829 TI - Retinal vascular caliber and extracranial carotid disease in patients with acute ischemic stroke: the Multi-Centre Retinal Stroke (MCRS) study. AB - BACKGROUND AND PURPOSE: Previous studies show that both retinal vascular caliber and carotid disease predict incident stroke in the general population, but the exact relationship between these 2 microvascular and macrovascular structural risk factors is unclear. We studied the relationship between retinal vascular caliber and carotid disease in patients presenting with acute ischemic stroke. METHODS: We conducted a cross-sectional study of patients with acute ischemic stroke recruited from 3 centers (Melbourne, Sydney, Singapore). The caliber of retinal arterioles and venules was measured from digital retinal photographs. Severe extracranial carotid disease was defined as stenosis >or=75% or occlusion determined by carotid Doppler using North American Symptomatic Carotid Endarterectomy Trial-based criteria. RESULTS: Among the 1029 patients with acute stroke studied, 7% of the population had severe extracranial carotid disease. Retinal venular caliber was associated with ipsilateral severe carotid disease (P<0.001 in multivariate models). Patients with wider retinal venular caliber were more likely to have severe ipsilateral carotid disease (multivariable adjusted OR, 3.81; 95% CI, 1.80 to 8.07, comparing the largest and smallest venular caliber quartiles). The retinal venular caliber-carotid disease association remained significant in patients with large artery stroke. CONCLUSIONS: In patients with acute stroke, retinal venular widening was strongly associated with ipsilateral severe extracranial carotid disease. Our findings suggest concomitant retinal and cerebral microvascular disease may be present in patients with carotid stenosis or occlusion disease. The pathogenesis of stroke due to carotid disease may thus be partially mediated by microvascular disease. PMID- 19815830 TI - Substantial observer variability in the differentiation between primary intracerebral hemorrhage and hemorrhagic transformation of infarction on CT brain imaging. AB - BACKGROUND AND PURPOSE: CT remains the most commonly used imaging technique in acute stroke but is often delayed after minor stroke. Interobserver reliability in distinguishing hemorrhagic transformation of infarction from intracerebral hemorrhage may depend on delays to CT but has not been reported previously despite the clinical importance of this distinction. METHODS: Initial CT scans with intraparenchymal hematoma from the first 1000 patients with stroke in the Oxford Vascular Study were independently categorized as intracerebral hemorrhage or hemorrhagic transformation of infarction by 5 neuroradiologists, both blinded and unblinded to clinical history. Thirty scans were reviewed twice. Agreement was quantified by the kappa statistic. RESULTS: Seventy-eight scans showed intraparenchymal hematoma. Blinded pairwise interrater agreements for a diagnosis of intracerebral hemorrhage ranged from kappa=0.15 to 0.48 with poor overall agreement (kappa=0.35; 95% CI, 0.15 to 0.54) even after unblinding (kappa=0.41; 0.21 to 0.60). Blinded intrarater agreements ranged from kappa=0.21 to 0.92. Lack of consensus after unblinding was greatest in patients scanned >or=24 hours after stroke onset (67% versus 25%, P=0.001) and in minor stroke (National Institutes of Health Stroke Scale or=24 hours after minor stroke and in 48% of all 30-day stroke survivors in whom reliable diagnosis would be expected to influence long-term management. CONCLUSIONS: Reliability of diagnosis of intraparenchymal hematoma on CT brain scan in minor stroke is poor, particularly if scanning is delayed. Immediate brain imaging is justified in patients with minor stroke. PMID- 19815831 TI - Circulating erythrocyte-derived microparticles are associated with coagulation activation in sickle cell disease. AB - BACKGROUND: Sickle cell disease is characterized by a hypercoagulable state as a result of multiple factors, including chronic hemolysis and circulating cell derived microparticles. There is still no consensus on the cellular origin of such microparticles and the exact mechanism by which they may enhance coagulation activation in sickle cell disease. DESIGN AND METHODS: In the present study, we analyzed the origin of circulating microparticles and their procoagulant phenotype during painful crises and steady state in 25 consecutive patients with sickle cell disease. RESULTS: The majority of microparticles originated from platelets (GPIIIa,CD61) and erythrocytes (glycophorin A,CD235), and their numbers did not differ significantly between crisis and steady state. Erythrocyte-derived microparticles strongly correlated with plasma levels of markers of hemolysis, i.e. hemoglobin (r=-0.58, p<0.001) and lactate dehydrogenase (r=0.59, p<0.001), von Willebrand factor as a marker of platelet/endothelial activation (r=0.44, p<0.001), and D-dimer and prothrombin fragment F1+2 (r=0.52, p<0.001 and r=0.59, p<0.001, respectively) as markers of fibrinolysis and coagulation activation. Thrombin generation depended on the total number of microparticles (r=0.63, p<0.001). Anti-human factor XI inhibited thrombin generation by about 50% (p<0.001), whereas anti-human factor VII was ineffective (p>0.05). The extent of factor XI inhibition was associated with erythrocyte-derived microparticles (r=0.50, p=0.023). CONCLUSIONS: We conclude that the procoagulant state in sickle cell disease is partially explained by the factor XI-dependent procoagulant properties of circulating erythrocyte-derived microparticles. PMID- 19815832 TI - Unimpaired terminal erythroid differentiation and preserved enucleation capacity in myelodysplastic 5q(del) clones: a single cell study. AB - BACKGROUND: Anemia is a characteristic of myelodysplastic syndromes, such as the rare 5q- syndrome, but its mechanism remains unclear. In particular, data are lacking on the terminal phase of differentiation of erythroid cells (enucleation) in myelodysplastic syndromes. DESIGN AND METHODS: We used a previously published culture model to generate mature red blood cells in vitro from human hematopoietic progenitor cells in order to study the pathophysiology of the 5q- syndrome. Our model enables analysis of cell proliferation and differentiation at a single cell level and determination of the enucleation capacity of erythroid precursors. RESULTS: The erythroid commitment of 5q(del) clones was not altered and their terminal differentiation capacity was preserved since they achieved final erythroid maturation (enucleation stage). The drop in red blood cell production was secondary to the decrease in the erythroid progenitor cell pool and to impaired proliferative capacity. RPS14 gene haploinsufficiency was related to defective erythroid proliferation but not to differentiation capacity. CONCLUSIONS: The 5q- syndrome should be considered a quantitative rather than qualitative bone marrow defect. This observation might open the way to new therapeutic concepts. PMID- 19815833 TI - Chronic hemolytic anemia due to novel alpha-globin chain variants: critical location of the mutation within the gene sequence for a dominant effect. PMID- 19815835 TI - Early cardiac iron overload in children with transfusion-dependent anemias. PMID- 19815834 TI - Improved survival in thalassemia major patients on switching from desferrioxamine to combined chelation therapy with desferrioxamine and deferiprone. PMID- 19815836 TI - Species-specific anticoagulant and mitogenic activities of murine protein S. AB - BACKGROUND: The protein C pathway down-regulates thrombin generation and promotes cytoprotection during inflammation and stress. In preclinical studies using models of murine injury (e.g., sepsis and ischemic stroke), murine protein S may be required because of restrictive species specificity. DESIGN AND METHODS: We prepared and characterized recombinant murine protein S using novel coagulation assays, immunoassays, and cell proliferation assays. RESULTS: Purified murine protein S had good anticoagulant co-factor activity for murine activated protein C, but not for human activated protein C, in mouse or rat plasma. In human plasma, murine protein S was a poor co-factor for murine activated protein C and had no anticoagulant effect with human activated protein C, suggesting protein S species specificity for factor V in addition to activated protein C. We estimated that mouse plasma contains 22+/-1 microg/mL protein S and developed assays to measure activated protein C co-factor activity of the protein S in murine plasma. Activated protein C-independent anticoagulant activity of murine protein S was demonstrable and quantifiable in mouse plasma, and this activity was enhanced by exogenous murine protein S. Murine protein S promoted the proliferation of mouse and human smooth muscle cells. The potency of murine protein S was higher for mouse cells than for human cells and similarly, human protein S was more potent for human cells than for mouse cells. CONCLUSIONS: The spectrum of bioactivities of recombinant murine protein S with mouse plasma and smooth muscle cells is similar to that of human protein S. However, in vitro and in vivo studies of the protein C pathway in murine disease models are more appropriately performed using murine protein S. This study extends previous observations regarding the remarkable species specificity of protein S to the mouse. PMID- 19815837 TI - Thalidomide decreases gelatinase production by malignant B lymphoid cell lines through disruption of multiple integrin-mediated signaling pathways. AB - BACKGROUND: Thalidomide and its analogs are effective agents in the treatment of multiple myeloma. Since gelatinases (matrix metalloproteinases-2 and -9) play a crucial role in tumor progression, we explored the effect of thalidomide on gelatinase production by malignant B lymphoid cell lines. DESIGN AND METHODS: We investigated the effect of therapeutic doses of thalidomide on integrin-mediated production of gelatinases by malignant B lymphoid cell lines by gelatin zymography, western-blot, reverse transcriptase polymerase chain reaction and invasive capacity through Matrigel-coated Boyden chambers. We also explored the effect of thalidomide on the activation status of the main signaling pathways involved in this process. RESULTS: Thalidomide strongly inhibited gelatinase production by B-cell lines and primary myeloma cells in response to fibronectin, the most efficient gelatinase inducer identified in lymphoid cells. Thalidomide disrupted integrin-mediated signaling pathways involved in gelatinase induction and release, such as Src and MAP-kinase ERK activation, resulting in decreased cell motility and invasiveness. Unexpectedly, treatment with thalidomide elicited an increase in fibronectin-induced Akt phosphorylation through phosphoinositide 3 kinase-independent pathways since thalidomide decreased fibronectin-induced phosphoinositide 3-kinase phosphorylation and reversed the inhibition of Akt phosphorylation achieved by the phosphoinositide 3-kinase inhibitors wortmannin and LY294002. CONCLUSIONS: Disruption of integrin-mediated signaling may be an important mechanism through which thalidomide and its analogs impair tumor cell interactions with the microenvironment. The unexpected effects of thalidomide on Akt activation indicate the need for further studies to elucidate whether the interference with Akt downstream effects would synergize with the anti-tumor activity of thalidomide. PMID- 19815838 TI - Prevalence and pathogenesis of anemia in inflammatory bowel disease. Influence of anti-tumor necrosis factor-alpha treatment. AB - BACKGROUND: Anemia is a common complication of inflammatory bowel disease, but its epidemiology may be changing due to earlier diagnosis and improved treatments. We investigated the prevalence and pathogenesis of anemia in patients with inflammatory bowel disease. DESIGN AND METHODS: In a cross-sectional study 263 out-patients with inflammatory bowel disease (165 with Crohn's disease, 98 with ulcerative colitis) were investigated. The influence of time from diagnosis, disease activity, inflammation and the status of iron and hematinic vitamins on the level of hemoglobin and prevalence of anemia were evaluated. In a second group of 27 patients with Crohn's disease, undergoing anti-tumor necrosis factor alpha treatment with infliximab because of refractory or fistulizing disease, we determined the effects of infliximab on disease activity, hemoglobin, serum erythropoietin levels, iron status and inflammation. RESULTS: In all, 104 of the 263 patients with inflammatory bowel disease were anemic. Age, gender and azathioprine treatment had no influence on anemia. The prevalence of anemia was highest at diagnosis (65%), decreased during the first 4 years after disease onset, and was stable thereafter. Active disease was associated with higher rates of anemia. At diagnosis most anemic patients had anemia of chronic disease; during follow-up iron deficiency and multifactorial forms of anemia became more prevalent. Eighteen of 27 patients undergoing treatment with infliximab were anemic; most of them had anemia of chronic disease. Infliximab reduced disease activity and improved anemia in 12 patients. This was mediated by an increased production of erythropoietin for the degree of anemia. In vitro infliximab increased the growth of erythroid progenitors from the peripheral blood of patients with active disease. Conclusions Anemia is a common problem in out patients with inflammatory bowel disease; the prevalence and severity of anemia are related to the activity of the bowel disorder. The pathogenesis of anemia changes during the course of the disease, with anemia of chronic disease having a major role at diagnosis and iron deficiency and multifactorial forms of anemia during follow-up. In patients requiring anti-tumor necrosis factor-alpha treatment, response to therapy improves erythropoiesis. PMID- 19815840 TI - Mobilization of PML/RARalpha negative peripheral blood stem cells with a combination of G-CSF and CXCR4 blockade in relapsed acute promyelocytic leukemia pre-treated with arsenic trioxide. PMID- 19815839 TI - Akt inhibitors induce apoptosis in chronic lymphocytic leukemia cells. AB - BACKGROUND: The phosphatidylinositol-3-kinase/Akt pathway has been described to be critical in the survival of chronic lymphocytic leukemia cells. In this study we analyzed the effect of two selective chemical inhibitors of Akt (Akti-1/2 and A-443654) on the survival of chronic lymphocytic leukemia cells. DESIGN AND METHODS: Using cytometry we studied the cytotoxic effects of Akt inhibitors on peripheral B and T lymphocytes from patients with chronic lymphocytic leukemia and from healthy donors. We studied the changes induced by Akti-1/2 and A-443654 at the mRNA level by performing reverse transcriptase multiplex ligation dependent probe amplification. We also studied the changes induced by both Akt inhibitors in some BCL-2 protein family members on chronic lymphocytic leukemia cells by western blotting. Moreover, we analyzed the cytotoxic effect of Akt inhibitors in patients' cells with deleted/mutated TP53. RESULTS: Both inhibitors induced apoptosis in chronic lymphocytic leukemia cells in a dose-dependent manner. Moreover, B cells from patients with chronic lymphocytic leukemia were more sensitive to Akt inhibitors than T cells from leukemic patients, and B or T cells from healthy donors. Survival factors for chronic lymphocytic leukemia cells, such as interleukin-4 and stromal cell-derived factor-1alpha, were not able to block the apoptosis induced by either Akt inhibitor. Akti-1/2 did not induce any change in the mRNA expression profile of genes involved in apoptosis, while A-443654 induced some changes, including an increase in NOXA and PUMA mRNA levels, suggesting the existence of additional targets for A-443654. Both inhibitors induced an increase in PUMA and NOXA protein levels, and a decrease in MCL-1 protein level. Moreover, Akti-1/2 and A-443654 induced apoptosis irrespective of TP53 status. CONCLUSIONS: These results demonstrate that Akt inhibitors induce apoptosis of chronic lymphocytic leukemia cells and might be a new therapeutic option for the treatment of chronic lymphocytic leukemia. PMID- 19815841 TI - I am amazed at how time flies, and that we are now reading the fifth issue of International Journal of Toxicology for 2009! PMID- 19815842 TI - Safety evaluation of a peptide product derived from sardine protein hydrolysates (valtyron). AB - The peptide product, Valtyron, is obtained via enzymatic hydrolysis of sardine muscle. Although the safety and efficacy of the sardine peptide product have been evaluated in human studies, sardine peptides have not been identified as the subject of toxicological testing. In this study, the sardine peptide product did not exhibit any mutagenic activity in Salmonella typhimurium or Escherichia coli WP2uvrA. Likewise, the sardine peptide product was not associated with clastogenic properties in mouse bone marrow cells in a micronucleus assay. An oral rat LD(50) value of greater than 10,000 mg per kilogram of body weight was determined for peptide alpha-1000, and in rats administered peptide alpha-1000 by gavage at levels up to 5000 mg per kilogram of body weight per day for 28 days, no compound-related differences were observed in standard toxicological parameters. The results of these studies support the safety of the sardine peptide product for use in food for human consumption as a dietary source of peptides available from sardines. PMID- 19815843 TI - Safety assessment of ProBiora3, a probiotic mouthwash: subchronic toxicity study in rats. AB - Streptococcus viridans are commensal bacteria that constitute a significant portion of the resident oral microflora. The objective of the present study is to investigate adverse effects, if any, of a blend of 3 natural strains, Streptococcus uberis KJ2, Streptococcus oralis KJ3, and Streptococcus rattus JH145 (probiotic mouthwash, ProBiora(3)). The blend is administered to rats orally once daily (5 days per week) at doses of 0, 10(6), or 10(9) colony-forming units of each strain for 14 weeks. No treatment-related adverse effects are observed in the physiological parameters during the study or in the evaluation of blood and tissue samples taken from the animals at the end. Results of an in vitro antibiotic susceptibility study demonstrate that all 3 ProBiora(3) strains are susceptible to commonly used therapeutic antibiotics. The results of these investigations reveal that the no-observed-adverse-effect level of the probiotic mouthwash is 2.16 x 10(9) colony-forming units per strain per kilogram of body weight per day, the highest dose used. PMID- 19815844 TI - Predictive modeling of a mixture of thyroid hormone disrupting chemicals that affect production and clearance of thyroxine. AB - Thyroid hormone (TH) disrupting compounds interfere with both thyroidal and extrathyroidal mechanisms to decrease circulating thyroxine (T(4)). This research tested the hypothesis that serum T(4) concentrations of rodents exposed to a mixture of both TH synthesis inhibitors (pesticides) and stimulators of T(4) clearance in the liver (polyhalogenated aromatic hydrocarbons, PHAHs) could be best predicted by an integrated addition model. Female Long-Evans rats, 23 days of age, were dosed with dilutions of a mixture of 18 PHAHs (2 dioxins, 4 dibenzofurans, and 12 PCBs, including dioxin-like and non-dioxin like PCBs) and a mixture of 3 pesticides (thiram, pronamide, and mancozeb) for four consecutive days. Serum was collected 24 hours after the last exposure and T(4) concentrations were measured by radioimmunoassay. Animals exposed to the highest dose of the mixture experienced a 45% decrease in serum T(4). Three additivity model predictions (dose addition, effect addition, and integrated addition) were generated based on single chemical data, and the results were compared. Effect addition overestimated the effect produced by the combination of all 21 chemicals. The results of the dose- and integrated-addition models were similar, and both provided better predictions than the effect-addition model. These results support the use of dose- and integrated additivity models in predicting the effects of complex mixtures. PMID- 19815846 TI - Primary DNA damage assessed with the comet assay and comparison to the absorbed dose of diagnostic X-rays in children. AB - The aim of this work is to assess DNA damage in peripheral blood lymphocytes of children prior to and following airway X-ray examinations of the chest using the alkaline comet assay and to compare data with the measured absorbed dose. Twenty children with pulmonary diseases, between the ages of 5 and 14 years, are assessed. Absorbed dose measurements are conducted for posterior-anterior projection on the forehead, thyroid gland, gonads, chest, and back. Doses are measured using thermoluminescent and radiophotoluminescent dosimetry systems. Differences between tail lengths, tail intensity, and tail moments as well as for the long-tailed nuclei before and after exposures are statistically significant and are dependent on the individual. The results demonstrate the usefulness of the comet assay as a measure of X-ray damage to lymphocytes in a clinical setting. Doses measured with both dosimeters show satisfactory agreement (0.01 mSv) and are suitable for dosimetric measurements in X-ray diagnostics. PMID- 19815845 TI - In vivo acute exposure to polychlorinated biphenyls: effects on free and total thyroxine in rats. AB - Hypothyroxinemia in rats has been well documented as a result of exposure to polychlorinated biphenyls (PCBs). Hypothetical mechanisms include induction of hepatic catabolic enzymes and cellular hormone transporters, and/or interference with plasma transport proteins. We hypothesized that if thyroxine displacement from transport proteins by PCBs occurs in vivo, it would result in increased free thyroxine (FT4). This study investigates the effects of a single oral dose of 2,2',4,4',5,5'-hexachlorobiphenyl (PCB 153 at 60 mg/kg) or 3,3',4,4',5,5' hexachlorobiphenyl (PCB 169 at 1 mg/kg) on rats at 28 or 76 days of age. Total thyroxine (TT4) and FT4 were measured at 0.5, 1, 2, 4, 8, 24, or 48 hours post dosing. Microsomal ethoxy- and pentoxy-resorufin-O-deethylase (EROD and PROD) activity and uridine diphosphoglucuronosyl transferase (UGT) activity were determined. No significant increase in TT4 or FT4 concentrations was seen at any time point. PCB 153 significantly decreased TT4 and FT4 in young and adult rats, with young rats showing a time-by-treatment interaction from 2 to 48 hours post dosing in serum FT4. With PCB 169 exposure, young rats showed a decrease in FT4 only, whereas adult rats showed decreases in TT4 only. Hepatic EROD and PROD activities were both dramatically increased following PCB 169 and 153, respectively. Uridine diphosphoglucuronosyl transferase activity was increased only after PCB 169 exposure. These data demonstrate that neither PCB 153 nor PCB169 increased FT4, which supports the conclusion that these PCBs do not displace thyroxine from serum TTR, or if it does occur, there is no subsequent increase in serum FT4 in vivo. PMID- 19815847 TI - Acute microinstillation inhalation exposure to sarin induces changes in respiratory dynamics and functions in guinea pigs. AB - This study investigates the toxic effects of sarin on respiratory dynamics following microinstillation inhalation exposure in guinea pigs. Animals are exposed to sarin for 4 minutes, and respiratory functions are monitored at 4 hours and 24 hours by whole-body barometric plethysmography. Data show significant changes in respiratory dynamics and function following sarin exposure. An increase in respiratory frequency is observed at 4 hours post exposure compared with saline controls. Tidal volume and minute volume are also increased in sarin-exposed animals 4 hours after exposure. Peak inspiratory flow increases, whereas peak expiratory flow increases at 4 hours and is erratic following sarin exposure. Animals exposed to sarin show a significant decrease in expiratory time and inspiratory time. End-inspiratory pause is unchanged whereas end-expiratory pause is slightly decreased 24 hours after sarin exposure. These results indicate that inhalation exposure to sarin alters respiratory dynamics and function at 4 hours, with return to normal levels at 24 hours post exposure. PMID- 19815849 TI - Use of archived specimens in evaluation of prognostic and predictive biomarkers. AB - The development of tumor biomarkers ready for clinical use is complex. We propose a refined system for biomarker study design, conduct, analysis, and evaluation that incorporates a hierarchal level of evidence scale for tumor marker studies, including those using archived specimens. Although fully prospective randomized clinical trials to evaluate the medical utility of a prognostic or predictive biomarker are the gold standard, such trials are costly, so we discuss more efficient indirect "prospective-retrospective" designs using archived specimens. In particular, we propose new guidelines that stipulate that 1) adequate amounts of archived tissue must be available from enough patients from a prospective trial (which for predictive factors should generally be a randomized design) for analyses to have adequate statistical power and for the patients included in the evaluation to be clearly representative of the patients in the trial; 2) the test should be analytically and preanalytically validated for use with archived tissue; 3) the plan for biomarker evaluation should be completely specified in writing before the performance of biomarker assays on archived tissue and should be focused on evaluation of a single completely defined classifier; and 4) the results from archived specimens should be validated using specimens from one or more similar, but separate, studies. PMID- 19815848 TI - Di (n-butyl) phthalate inhibits testosterone synthesis through a glucocorticoid mediated pathway in rats. AB - The present study focused on investigating whether the inhibitory effect of di (n butyl) phthalate (DBP) on testosterone (T) biosynthesis was mediated by the glucocorticoid (GC) pathway in prepubertal male rats and T production after the exposure to DBP ceased. Prepubertal male rats were administered DBP in corn oil orally at 0, 250, 500, 1000, and 2000 mg/kg daily for 30 days. Serum T and GC were measured by radioimmunoassay and enzyme-linked immunosorbent assay, respectively. The responses, including glucocorticoid receptor (GR), type I 11beta-hydroxysteroid dehydrogenase (11beta-HSD1), and steroidogenesis acute regulatory protein (StAR) in the testes tissues, were determined by Western blotting and reverse transcriptase PCR. DBP exposure resulted in testicular toxicity, such as seminiferous tubule degeneration and a decrease in the number of spermatogenic cells. T was decreased and GC was increased in a DBP concentration-dependent manner in the exposure group. The expression of GR and 11beta-HSD1 was significantly increased, with an associated decrease in expression of StAR. Neither the expression of the GR nor 11beta-HSD1 and StAR were statistically significantly different in the postexposure group compared with the control. However, the weight and morphology of the testes did not recover in the postexposure group. These data suggest that DBP inhibits testosterone production through a GC-mediated pathway in prepubertal male rats, and after exposure to DBP ceases, testosterone biosynthesis returns. PMID- 19815850 TI - MUC2 mucin is a major carrier of the cancer-associated sialyl-Tn antigen in intestinal metaplasia and gastric carcinomas. AB - Changes in mucin protein expression and in glycosylation are common features in pre-neoplastic lesions and cancer and are therefore used as cancer-associated markers. De novo expression of intestinal mucin MUC2 and cancer-associated sialyl Tn antigen are frequently observed in intestinal metaplasia (IM) and gastric cancer. However, despite that these antigens often co-localize, MUC2 has not been demonstrated to be a carrier of sialyl-Tn. By using the in situ proximity ligation assay (in situ PLA), we herein could show that MUC2 is a major carrier of the sialyl-Tn antigen in all IM cases and in most gastric carcinoma cases. The requirement by in situ PLA for the presence of both antigens in close proximity increases the selectivity compared to measurement of co-localization, as determined by immunohistochemistry. Identification of the mucin which is the carrier of a carbohydrate structure offers unique advantages for future development of more accurate diagnostic and prognostic markers. PMID- 19815851 TI - Vascular remodeling: in situ vs. ex vivo. PMID- 19815852 TI - Vascular remodeling arterioles: plasticity of the vessel wall. PMID- 19815854 TI - Physiological signaling specificity by protein tyrosine phosphatases. AB - Protein tyrosine phosphatases (PTPs) are now recognized to be involved in a multitude of signaling events that control fundamental biological processes such as cell growth, differentiation, apoptosis, and cell movement. PTPs, which were initially thought to be less discriminating in their actions compared with their protein tyrosine kinase counterparts, are now known to regulate these various biological processes in a precise manner. This review will focus on the concept that PTPs exhibit remarkable signaling specificity through intrinsic differences between their PTP domains and through various modes of regulation that endows them with the capacity to promote unique physiological responses. PMID- 19815856 TI - Role of extracellular adenosine in acute lung injury. AB - Acute lung injury (ALI) is a lung disease characterized by pulmonary edema and severe hypoxia. The past decade hosted a search for endogenous mechanisms controlling lung inflammation and pulmonary edema during ALI. As such, recent evidence indicates extracellular adenosine in orchestrating the resolution of pulmonary edema and inflammation during ALI. PMID- 19815855 TI - Oligodendrocyte development and myelin biogenesis: parsing out the roles of glycosphingolipids. AB - The myelin sheath is an extension of the oligoddendrocyte (OL) plasma membrane enriched in lipids that ensheaths the axons of the central and peripheral nervous system. Here, we review the involvement of glycosphingolipids in myelin/OL functions, including the regulation of OL differentiation, lipid raft-mediated trafficking and signaling, and neuron-glia interactions. PMID- 19815857 TI - Potassium ion movement in the inner ear: insights from genetic disease and mouse models. AB - Sensory transduction in the cochlea and vestibular labyrinth depends on fluid movements that deflect the hair bundles of mechanosensitive hair cells. Mechanosensitive transducer channels at the tip of the hair cell stereocilia allow K(+) to flow into cells. This unusual process relies on ionic gradients unique to the inner ear. Linking genes to deafness in humans and mice has been instrumental in identifying the ion transport machinery important for hearing and balance. Morphological analysis is difficult in patients, but mouse models have helped to investigate phenotypes at different developmental time points. This review focuses on cellular ion transport mechanisms in the stria vascularis that generate the major electrochemical gradients for sensory transduction. PMID- 19815861 TI - Cerebral malaria: a new way forward with magnetic resonance imaging (MRI). AB - Magnetic resonance studies offer a new way through the impasse that now seems to block further progress in disentangling the pathogenesis and improving the treatment of cerebral malaria, a catastrophic neurologic complication of infection with Plasmodium falciparum. The underlying mechanisms responsible for coma in cerebral malaria are still unknown and the relative contributions of the microvascular sequestration of infected erythrocytes, the inflammatory response to P. falciparum, disordered hemostasis, and other factors remain controversial. For more than a century, neuropathologic studies have provided the basis for concepts of causation of cerebral malaria. Magnetic resonance techniques now offer non-invasive means of determining essential anatomic, metabolic, biochemical, and functional features of the brain in patients with cerebral malaria during life that could transform our understanding of the pathogenesis of cerebral malaria and lead to the development of new neuroprotective treatments. PMID- 19815862 TI - Peripheral rim enhancement in tuberculous mediastinal lymph nodes. PMID- 19815863 TI - Cervical schistosomiasis as a risk factor of cervical uterine dysplasia in a traveler. AB - Female genital schistosomiasis (FGS) may be under-recognized in endemic areas as a cause of cervical dysplasia, neoplasia, infertility, and as a facilitator of the transmission of HIV. To the best of our knowledge, few cases of FGS mimicking neoplasia have been reported in travelers. We report a clinical case of a 34-year old white woman who presented with a severe cervical dysplasia, without any features of human papilloma virus infection, 2 years after bathing in a waterfall, a source of schistosomiasis, in Mali. Schistosomes eggs were found on the conization. Management included conization and medical treatment, resulting in a full clinical and histologic recovery. FGS should be kept in mind as a possible cause of cervical dysplasia in endemic areas. Medical treatment with praziquantel improves this condition. PMID- 19815859 TI - Hospitalized patients with 2009 H1N1 influenza in the United States, April-June 2009. AB - BACKGROUND: During the spring of 2009, a pandemic influenza A (H1N1) virus emerged and spread globally. We describe the clinical characteristics of patients who were hospitalized with 2009 H1N1 influenza in the United States from April 2009 to mid-June 2009. METHODS: Using medical charts, we collected data on 272 patients who were hospitalized for at least 24 hours for influenza-like illness and who tested positive for the 2009 H1N1 virus with the use of a real-time reverse-transcriptase-polymerase-chain-reaction assay. RESULTS: Of the 272 patients we studied, 25% were admitted to an intensive care unit and 7% died. Forty-five percent of the patients were children under the age of 18 years, and 5% were 65 years of age or older. Seventy-three percent of the patients had at least one underlying medical condition; these conditions included asthma; diabetes; heart, lung, and neurologic diseases; and pregnancy. Of the 249 patients who underwent chest radiography on admission, 100 (40%) had findings consistent with pneumonia. Of the 268 patients for whom data were available regarding the use of antiviral drugs, such therapy was initiated in 200 patients (75%) at a median of 3 days after the onset of illness. Data suggest that the use of antiviral drugs was beneficial in hospitalized patients, especially when such therapy was initiated early. CONCLUSIONS: During the evaluation period, 2009 H1N1 influenza caused severe illness requiring hospitalization, including pneumonia and death. Nearly three quarters of the patients had one or more underlying medical conditions. Few severe illnesses were reported among persons 65 years of age or older. Patients seemed to benefit from antiviral therapy. PMID- 19815864 TI - Vestibular-evoked myogenic potential (VEMP) in the evaluation of schistosomal myeloradiculopathy. AB - Schistosomal myeloradiculopathy (SMR) is the most severe and disabling form of schistosomiasis. The diagnosis is based on clinical, laboratory, and image data. Vestibular-evoked myogenic potential (VEMP) is a neurophysiologic test that assesses the vestibulospinal pathway through acoustic or galvanic stimuli. The aim of this study was to evaluate cervical spinal abnormalities in patients with SMR. Fifty-two subjects were evaluated, of whom 29 had SMR and 30 did not (normal control). Normal VEMP was observed in all volunteers without SMR. Abnormal VEMP was recorded in 34% of the group with SMR. After treatment, abnormal VEMP was found in 80% of those with persistent neurologic abnormalities. VEMP is a functional test, and the alteration may precede image abnormalities. This procedure may be useful for early diagnosis of schistosomal cervical spinal cord involvement. PMID- 19815865 TI - Co-Infection of Leishmania (Viannia) braziliensis and HIV: report of a case of mucosal leishmaniasis in Cochabamba, Bolivia. AB - We describe the first case of Leishmania/HIV co-infection reported in Bolivia. Initially hospitalized with a diagnosis of pneumonia and bronchitis, the patient had numerous cutaneous and mucosal lesions caused by Leishmania (Viannia) braziliensis. The patient was also diagnosed as severely immunocompromised because of HIV infection. PMID- 19815866 TI - Tegumentary leishmaniasis as the cause of immune reconstitution inflammatory syndrome in a patient co-infected with human immunodeficiency virus and Leishmania guyanensis. AB - We report a case of immune reconstitution inflammatory syndrome (IRIS) in a 32 year-old man infected with human immunodeficiency virus and Leishmania guyanensis. Three months after initiation of highly active anti-retroviral therapy (HAART), the patient had disseminated cutaneous leishmaniasis and started anti-leishmanial therapy. The patient's leishmaniasis manifestations during HAART ranged form an anergic response (46 CD4+ T cells/microL) to a disseminated cutaneous leishmaniasis (112 CD4+ T cells/microL). Eight weeks later (168 CD4+ T cells/microL, skin biopsy specimens showed inflammatory infiltrates with no detectable amastigotes. The patient then became comatose. Prednisone therapy (60 mg/day) was initiated with a significant improvement within 48 hours. Three months later (CD4+ T cell count = 184 cell/microL), localized, classic, cutaneous leishmaniasis developed in the patient and anti-leishmanial treatment was re introduced. On that occasion, frequency of T regulatory cells was 1.82% of all CD4+ cells. Our data suggest a pivotal role for CD4+ T cells in the onset of IRIS and lesion ulceration and their association with a low frequency of T regulatory cells. PMID- 19815867 TI - Molecular epidemiology of American tegumentary leishmaniasis in Panama. AB - American tegumentary leishmaniasis is an increasing public health problem in Panama. This study describes the clinical characteristics and the molecular epidemiology of leishmaniasis in Panama over a 5-year period (2004-2008). Additionally, we applied a polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP)-based assay to identify Leishmania species in clinical isolates, skin scrapings, and sandflies specimens. Whereas 60.3% of cases were detected with conventional parasitologic techniques (smear or in vitro culture), the PCR detected 72% positive patients. Our clinical-epidemiologic data corroborate the high incidence of L. (Viannia) panamensis and provide evidence of peridomestic and/or domestic transmission. Mucosal involvement was observed in 4.2% of the patients. The overall natural infection rate with Leishmania in 103 pools of sandflies was 0.46%. Lutzomyia gomezi and Lutzomya panamensis were the prevalent species incriminated as vectors at the capture sites in central Panama. This study contributes to a better knowledge of the current epidemiology of tegumentary leishmaniasis in Panama. PMID- 19815868 TI - Vector incrimination of sand flies in the most important visceral leishmaniasis focus in Iran. AB - The prevalence, host preference, and rate of Leishmania spp. infection of sand fly species are important parameters for incrimination of parasite vectors. We applied polymerase chain reaction (PCR)-based and enzyme-linked immunosorbent assay (ELISA) methods to detect Leishmania spp. parasites and blood meals within individual sand flies in the most important visceral leishmaniasis (VL) focus in northwestern Iran. Leishmania spp. minicircles (kinetoplast DNA) were found in 14 (0.9%) of 1,569 female specimens. Sequence analysis of 650 basepairs of an internal transcribed spacer ribosomal DNA gene identified L. infantum/L. donovani in 12 specimens and L. adleri-like parasites in 2 specimens. Nine (64.3%) of 14 of the Leishmania spp.-positive sand flies were Phlebotomus perfeliewi transcaucasicus. Blood meal identification of host DNA within sand flies by PCR based and ELISA methods showed that 30% and 28%, respectively, were positive for human blood. Results of this study showed that P. perfeliewi transcaucasicus is the most prevalent, infected, and anthropophagic sand fly and plays a major role in VL transmission in the region studied. PMID- 19815860 TI - Critical care services and 2009 H1N1 influenza in Australia and New Zealand. AB - BACKGROUND: Planning for the treatment of infection with the 2009 pandemic influenza A (H1N1) virus through health care systems in developed countries during winter in the Northern Hemisphere is hampered by a lack of information from similar health care systems. METHODS: We conducted an inception-cohort study in all Australian and New Zealand intensive care units (ICUs) during the winter of 2009 in the Southern Hemisphere. We calculated, per million inhabitants, the numbers of ICU admissions, bed-days, and days of mechanical ventilation due to infection with the 2009 H1N1 virus. We collected data on demographic and clinical characteristics of the patients and on treatments and outcomes. RESULTS: From June 1 through August 31, 2009, a total of 722 patients with confirmed infection with the 2009 H1N1 virus (28.7 cases per million inhabitants; 95% confidence interval [CI], 26.5 to 30.8) were admitted to an ICU in Australia or New Zealand. Of the 722 patients, 669 (92.7%) were under 65 years of age and 66 (9.1%) were pregnant women; of the 601 adults for whom data were available, 172 (28.6%) had a body-mass index (the weight in kilograms divided by the square of the height in meters) greater than 35. Patients infected with the 2009 H1N1 virus were in the ICU for a total of 8815 bed-days (350 per million inhabitants). The median duration of treatment in the ICU was 7.0 days (interquartile range, 2.7 to 13.4); 456 of 706 patients (64.6%) with available data underwent mechanical ventilation for a median of 8 days (interquartile range, 4 to 16). The maximum daily occupancy of the ICU was 7.4 beds (95% CI, 6.3 to 8.5) per million inhabitants. As of September 7, 2009, a total of 103 of the 722 patients (14.3%; 95% CI, 11.7 to 16.9) had died, and 114 (15.8%) remained in the hospital. CONCLUSIONS: The 2009 H1N1 virus had a substantial effect on ICUs during the winter in Australia and New Zealand. Our data can assist planning for the treatment of patients during the winter in the Northern Hemisphere. PMID- 19815869 TI - Sensitive, specific, and rapid detection of Leishmania donovani DNA by loop mediated isothermal amplification. AB - We have applied a loop-mediated isothermal amplification (LAMP) technique to detect Leishmania donovani DNA. The LAMP technique detected 1 fg of L. donovani DNA, which was 10-fold more sensitive than a conventional polymerase chain reaction (PCR). All nested PCR-positive blood samples from visceral leishmaniasis patients were positive with the LAMP technique, and DNA samples from L. infantum, L. major, L. mexicana, L. tropica, L. braziliensis, Plasmodium falciparum, and healthy humans were negative with the LAMP technique. The advantages of the LAMP method are its shorter reaction time, a lack of requirement of sophisticated equipment, and visual judgment of positivity based on the turbidity of reaction mixture. Our LAMP technique can be a better alternative to a conventional PCR, especially under field conditions. PMID- 19815870 TI - Aspergillus flavus brain abscesses associated with hepatic amebiasis in a non neutropenic man in Senegal. AB - A non-neutropenic man living in Senegal was repatriated to France for liver amebic abscesses associated with brain abscesses presumed to be of amebic origin. Surprisingly, the post-mortem examinations of brain abscesses showed Aspergillus flavus. The route of infection by A. flavus in this particular context is discussed. PMID- 19815871 TI - Adaptation of a Thai multidrug-resistant C2A clone of Plasmodium falciparum to Aotus monkeys and its preliminary in vivo antimalarial drug efficacy-resistance profile. AB - A multidrug-resistant (MDR) clone of Plasmodium falciparum (C2A) from Thailand was adapted through serial passage to Aotus monkeys. During adaptation, the parasite showed resistance to a single 20 or 40 mg/kg oral dose of mefloquine (MQ). Infection was only cured when MQ was administered orally at 40 mg/kg once in combination with intravenous artesunic acid at 20 mg/kg for 3 days. Similarly, the parasite clone was found to be resistant to quinine, failing at 20 mg/kg orally for 5 days in combination with an experimental dihydrofolate reductase (DHFR) inhibitor (WR297608) at 10, 20, or 40 mg/kg orally for 3 days, and with atovaquone/proguanil at 25 mg/kg for 3 days. This new model will allow in vivo testing of new antimalarial compounds or their combinations against a currently circulating MDR P. falciparum strain. PMID- 19815872 TI - Severe imported malaria in adults: retrospective study of 20 cases. AB - Severe imported malaria is an important problem in many countries in which this disease is not endemic. This retrospective study describes the characteristics of 20 adults with severe imported malaria admitted to our intensive care unit from 1991 through 2007. All episodes were caused by Plasmodium falciparum and all patients had returned from sub-Saharan Africa, except for one transfusion recipient. All persons were considered non-immune, and none had taken appropriate chemoprophylaxis. The median time between the initiation of symptoms and the diagnosis was seven days. Five patients died (mortality rate = 25%). A higher frequency of unrousable coma and acidosis and a higher median Apache II score at admission was noted in the persons who died. Mortality by severe malaria remains high despite high quality management, which highlights the importance of chemoprophylaxis and early diagnosis and treatment. PMID- 19815873 TI - Biological resistance of hydroxychloroquine for Plasmodium vivax malaria in the Republic of Korea. AB - The Republic of Korea (ROK) Army instituted a vivax malaria chemoprophylaxis program (hydroxychloroquine [HCQ] 400 mg per week) in 1997 that was expanded to nearly 200,000 soldiers by 2007, raising concerns for the emergence of drug resistant vivax malaria. Therefore, a study of whole blood HCQ concentrations for all malaria patients admitted to four ROK Army hospitals was conducted from June through September 2007. For all 142 vivax malaria patients enrolled, fevers returned to normal by Day 3 post-treatment and all thin blood films were negative for parasites by Day 7. Pre-treatment whole blood concentrations of HCQ for 14 patients were > 100 ng/mL. Eight of the patients were enrolled in the ROK Army chemoprophylaxis program that reported taking HCQ as directed, with the last pill taken > or = 4 days before diagnosis. Although there was no evidence of clinical resistance, chemoprophylaxis data indicates the biological resistance or tolerance to HCQ in ROK. PMID- 19815874 TI - Resurgence of Plasmodium vivax malaria in the Republic of Korea during 2006-2007. AB - Plasmodium vivax malaria, which re-emerged in the Republic of Korea (ROK) in 1993, had decreased since 2001. However, case numbers began to increase again in 2005. The number of cases rose 54.0% in 2006, but the rate of increase slowed down in 2007. Among the total of 4,206 cases of P. vivax malaria during 2006 2007, 756 cases (18.0%) were ROK military personnel, 891 cases (21.2%) were veterans, and 2,559 cases (60.8%) were civilians. The rapid increase during this period was mostly contributed by the western part of the malaria-risk areas that is under the influence of adjacent North Korea. Local transmission cases in ROK have also increased gradually and the transmission period seemingly became longer. Chemoprophylaxis in the military should be re-assessed in view of chloroquine-resistance. Continuous surveillance and monitoring are warranted to prevent further expansion of P. vivax malaria caused by climate change in ROK. PMID- 19815875 TI - Assessing the impact of indoor residual spraying on malaria morbidity using a sentinel site surveillance system in Western Uganda. AB - A single round of indoor residual spraying (IRS) using lambda-cyhalothrin was implemented in a district of Uganda with moderate transmission intensity in 2007. Individual patient data were collected from one health facility within the district 8 months before and 16 months after IRS. There was a consistent decrease in the proportion of patients diagnosed with clinical malaria after IRS for patients < 5 and > 5 years of age (52% versus 26%, P < 0.001 and 36% versus 23%, P < 0.001, respectively). There was a large decrease in the proportion of positive blood smears in the first 4 months after IRS for patients < 5 (47% versus 14%, P < 0.001) and > 5 (26% versus 9%, P < 0.001) years of age, but this effect waned over the subsequent 12 months. IRS was effective in reducing malaria morbidity, but this was not sustained beyond 1 year for the proportion of blood smears read as positive. PMID- 19815877 TI - Non-destructive determination of age and species of Anopheles gambiae s.l. using near-infrared spectroscopy. AB - Determining malaria vector species and age is crucial to measure malaria risk. Although different in ecology and susceptibility to control, the African malaria vectors Anopheles gambiae sensu stricto and An. arabiensis are morphologically similar and can be differentiated only by molecular techniques. Furthermore, few reliable methods exist to estimate the age of these vectors, which is a key predictor of malaria transmission intensity. We evaluated the use of near infrared spectroscopy (NIRS) to determine vector species and age. This non destructive technique predicted the species of field-collected mosquitoes with approximately 80% accuracy and predicted the species of laboratory-reared insects with almost 100% accuracy. The relative age of young or old females was predicted with approximately 80% accuracy, and young and old insects were predicted with > or = 90% accuracy. For applications where rapid assessment of the age structure and species composition of wild vector populations is needed, NIRS offers a valuable alternative to traditional methods. PMID- 19815876 TI - Pharmacokinetic profiles of artesunate after single intravenous doses at 0.5, 1, 2, 4, and 8 mg/kg in healthy volunteers: a phase I study. AB - The pharmacokinetics of good manufacturing process injection of artesunate (AS) were evaluated after single doses at 0.5, 1, 2, 4, and 8 mg/kg with a 2-minute infusion in 40 healthy subjects. Drug concentrations were analyzed by validated liquid chromatography and mass spectrometry system (LC-MS/MS) procedures. The drug was immediately converted to dihydroartemisinin (DHA), with elimination half lives ranging 0.12-0.24 and 1.15-2.37 hours for AS and DHA, respectively. Pharmacokinetic model-dependent analysis is suitable for AS, whereas DHA fits both model-dependent and -independent methods. Although DHA concentration was superior to that of AS with a 1.12-1.87 ratio of area under the curve (AUC)(DHA/AS), peak concentration of AS was much higher than that of DHA, with a 2.80- to 4.51-fold ratio of peak concentration (C(max AS/DHA)). Therefore, AS effectiveness has been attributed not only to its rapid hydrolysis to DHA, but also to itself high initial C(max). PMID- 19815878 TI - Statins fail to improve outcome in experimental cerebral malaria and potentiate Toll-like receptor-mediated cytokine production by murine macrophages. AB - Cerebral malaria is responsible for a large proportion of the estimated one million deaths caused by Plasmodium falciparum malaria annually. This disease is associated with excessive pro-inflammatory cytokine production resulting from dysregulated host responses to infection. On the basis of reports indicating potent activity against host-mediated inflammatory disorders such as sepsis, we examined the activity of statins (3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors) on malaria-associated inflammation in vivo and in vitro. Simvastatin failed to improve survival or alter parasitemia in C57BL/6 mice infected with Plasmodium berghei ANKA, an experimental model of cerebral malaria. In vitro statin treatment potentiated production of tumor necrosis factor and interleukin 6 by murine peritoneal macrophages in response to P. falciparum glycosylphosphatidyl inositol, a Toll-like receptor 2 (TLR2) ligand. Statin treatment also potentiated pro-inflammatory cytokine production stimulated by a panel of TLR2 and TLR4 ligands. Our results indicate that statins fail to confer protection in experimental cerebral malaria and potentiate TLR-mediated pro inflammatory cytokine production by primary murine macrophages. PMID- 19815879 TI - Hemostatic changes in Vietnamese children with mild dengue correlate with the severity of vascular leakage rather than bleeding. AB - The mechanisms underlying the bleeding manifestations and coagulopathy associated with dengue remain unclear, in part because of the focus of much previous work on severe disease without an appropriate comparison group. We describe detailed clinical and laboratory profiles for a large group of children with dengue of all severities, and a group with similar non-dengue febrile illnesses, all followed prospectively from early presentation through to recovery. Among the dengue infected patients but not the controls, thrombocytopenia, increased partial thromboplastin times and reduced fibrinogen concentrations were apparent from an early stage, and these abnormalities correlated strongly with the severity and timing of vascular leakage but not bleeding. There was little evidence of procoagulant activation. The findings do not support a primary diagnosis of disseminated intravascular coagulation to explain the intrinsic coagulopathy. An alternative biologically plausible hypothesis is discussed. PMID- 19815880 TI - Effect of peridomestic environments on repeated infestation by preadult Aedes aegypti in urban premises in Nha Trang City, Vietnam. AB - To determine the effect of peridomestic environments on Aedes aegypti infestation in urban premises, we conducted two consecutive surveys in the hot-dry and cool wet seasons. Most Ae. aegypti pupae (79%) were recovered from premises where preadult forms (larvae and/or pupae) had been detected in both surveys. Hence, repeated infestation appears to be a useful parameter to identify premises associated with a high potential risk of dengue transmission. Multivariate analysis revealed that not only the persistent presence of containers discarded outdoors, wells, large plastic buckets, jars, and concrete toilet basins in the premises (adjusted odds ratios [aORs] = 63.3, 23.3, 22.5, 6.6, and 5.6, respectively) but also the presence of six or more residents was significantly associated with repeated infestation (aOR = 6.1). Premises with six or more residents along with specific container types from which a large number of pupae were recovered should be targeted in dengue-control programs. PMID- 19815881 TI - Feeding and defecation patterns of Rhodnius nasutus (Hemiptera; Reduviidae), a triatomine native to an area endemic for Chagas disease in the state of Ceara, Brazil. AB - The importance of Rhodnius nasutus in the transmission of Chagas disease in northeastern Brazil was investigated regarding feeding and defecation patterns of this triatomine under laboratory conditions. An average of 30 samples were studied for each instar, from fourth-instar nymphs onward. On average, 86.4% started feeding after less than 10 minutes. In terms of the duration of feeding, 53.3% of fourth instar nymphs, 81.9% of fifth-instar nymphs, 21.9% of males, and 36.7% of females fed for more than 15 minutes. In all groups, there were insects that defecated and urinated during feeding; adult males defecated the most and fourth instar nymphs defecated the least. The results demonstrate that R. nasutus may be considered an efficient T. cruzi vector because it avidly searches for a food source, has a lengthy feeding time with low probability of interruption during feeding, and achieves a high percentage of engorgement. PMID- 19815882 TI - Molecular epidemiology of Chagas disease in the wild transmission cycle: the evaluation in the sylvatic vector Mepraia spinolai from an endemic area of Chile. AB - The sylvatic transmission cycle of Chagas disease in Chile is composed of wild mammals and insects of the genus Mepraia. We determined infection rates and Trypanosoma cruzi genotypes in Mepraia spinolai. We collected 227 insects from two ecologically contrasting areas to assess T. cruzi infection. Polymerase chain reaction (PCR)-amplified minicircle DNAs were characterized by Southern blot and hybridization tests with genotype-specific probes. Infection in insects from the more fertile area was almost 2-fold higher than in the poorer area. The genotype TCI was the most prevalent and other genotypes such as TCIIb, TCIId, and TCIIe were found at lower rates. The areas differed in their genotype distribution but not in their genotype diversity. We suggest that the difference in abundance and richness of mammals between the areas may be producing the detected infection levels in vectors. Our results are compared with those reported for mammals from the same area. PMID- 19815883 TI - Clinical course of monoclonal and oligoclonal gammopathies in patients infected with Orientia tsutsugamushi. AB - Although we have encountered many cases of monoclonal gammopathies (MGs) or oligoclonal gammopathies (OGs) in patients with Orientia tsutsugamushi infections, the clinical course of MG/OG associated with these infections has rarely been reported. We serially monitored 18 cases of MG/OG that appeared in the acute phase or early convalescent phase of scrub typhus. All MGs/OGs associated with infection with O. tsutsugamushi disappeared 7-15 weeks after treatment with doxycycline. In three cases of scrub typhus, MGs/OGs appeared before seropositive conversion (presence of antibodies against O. tsutsugamushi) and disappeared before seronegative conversion. Understanding our cases may be helpful for relevant clinical counseling and making appropriate medical decisions to avoid unnecessary invasive or cost-intensive diagnostic procedures for MGs/OGs in scrub typhus, especially in disease-endemic areas. We also suggest that 2-4 months after antibiotic treatment might be the optimal follow-up time to observe the disappearance of MGs/OGs associated with O. tsutsugamushi infections. PMID- 19815885 TI - Crimean-Congo hemorrhagic fever virus as a nosocomial pathogen in Iran. AB - Crimean-Congo hemorrhagic fever (CCHF) is a viral disease with several different modes of transmission. We describe the manifestations, outcome, and likely modes of transmission for three nosocomial cases. All three cases were healthcare workers (two men and one woman). They had fever, myalgia, and petechia. Disseminated intravascular coagulation resulted in the death occurred in the woman. Because this disease is manifested with non-specific influenza-like symptoms, diagnosis can be difficult. Data for these patients can be used to investigate airborne or sexual transmission of this virus, although neither route was substantiated for these patients. Use of universal precautions and early case detection are the most helpful strategy for preventing nosocomial transmission of CCHF. PMID- 19815884 TI - Development of an Alamar Blue viability assay in 384-well format for high throughput whole cell screening of Trypanosoma brucei brucei bloodstream form strain 427. AB - There is an urgent need for new compounds for the drug development pipeline for treatment of patients with African sleeping sickness. One approach for identifying such compounds is by high throughput screening (HTS) of compound collections. For time and cost considerations, there is a need for the development of an assay that uses at least 384-well formats. To our knowledge, there are currently no viability assays for whole cell screening of trypanosomes in the 384-well plate format. We have developed and optimized an Alamar Blue viability assay in a 384-well format for Trypanosoma brucei brucei bloodstream form strain 427 (BS427). The assay had a Z' > 0.5 and tolerated a final dimethyl sulfoxide concentration of 0.42%. Drug sensitivity was compared with those reported from previously developed 96-well methods and was found to be comparable. The sensitivity and cost benefit of the Alamar Blue assay make it an excellent candidate for HTS application. PMID- 19815886 TI - Development of field-based real-time reverse transcription-polymerase chain reaction assays for detection of Chikungunya and O'nyong-nyong viruses in mosquitoes. AB - Chikungunya (CHIK) and O'nyong-nyong (ONN) are important emerging arthropod-borne diseases. Molecular diagnosis of these two viruses in mosquitoes has not been evaluated, and the effects of extraneous mosquito tissue on assay performance have not been tested. Additionally, no real-time reverse transcription-polymerase chain reaction (RT-PCR) assay exists for detecting ONN virus (ONNV) RNA. We describe the development of sensitive and specific real-time RT-PCR assays for detecting CHIK and ONN viral RNA in mosquitoes, which have application for field use. In addition, we compared three methods for primer/probe design for assay development by evaluating their sensitivity and specificity. This comparison resulted in development of virus-specific assays that could detect less than one plaque-forming unit equivalent of each of the viruses in mosquitoes. The use of these assays will aid in arthropod-borne disease surveillance and in the control of the associated diseases. PMID- 19815887 TI - Comparative efficacy of BioUD to other commercially available arthropod repellents against the ticks Amblyomma americanum and Dermacentor variabilis on cotton cloth. AB - BioUD is an arthropod repellent that contains the active ingredient 2-undecanone originally derived from wild tomato plants. Repellency of BioUD was compared with five commercially available arthropod repellents against the ticks Amblyomma americanum (L.) and Dermacentor variabilis Say in two-choice bioassays on treated versus untreated cotton cheesecloth. Overall mean percentage repellency against both species was greatest for and did not differ significantly between BioUD (7.75% 2-undecanone) and products containing 98.1% DEET, 19.6% IR3535, and 30% oil of lemon eucalyptus. Products containing 5% and 15% Picaridin and 0.5% permethrin were also repellent compared with untreated controls but to a lesser degree than BioUD. The four most active repellents at the same concentrations used before were directly compared in head-to-head bioassays on cotton cheesecloth. BioUD provided significantly greater overall mean percentage repellency than IR3535 for A. americanum and D. variabilis. BioUD was significantly more repellent than oil of lemon eucalyptus for A. americanum but did not differ significantly in repellency against D. variabilis. No statistically significant difference in overall mean percentage repellency was found between BioUD and DEET for A. americanum or D. variabilis. In a 7-week time course bioassay, BioUD applied to cotton cheesecloth and held at room temperature provided 5 weeks of > 90% repellency against A. americanum. PMID- 19815888 TI - Seroprevalence of Q fever in the United States, 2003-2004. AB - We performed serum testing for IgG antibodies against Coxiella burnetii (phase I and phase II) and analyzed questionnaire data from 4,437 adults > or = 20 years of age who participated in the National Health and Nutrition Examination Survey 2003-2004 survey cycle. National Q fever seroprevalence was determined by enzyme linked immunosorbent assay and confirmed by using immunofluorescent antibody testing. Overall seroprevalence for Coxiella burnetii was 3.1% (95% confidence interval [CI] = 2.1-4.3%) among 4,437 adults > or = 20 years of age. Coxiella burnetii age-adjusted antibody prevalence was higher for men than for women (3.8%, 95% CI = 2.7-5.2% versus 2.5%, 95% CI = 1.5-3.7%, respectively, P < 0.05). Mexican Americans had a significantly higher antibody prevalence (7.4%, 95% CI = 6.6-8.3%) than either non-Hispanic whites (2.8%, 95% CI = 1.7-4.3%) or non Hispanic blacks (1.3%, 95% CI = 0.6-2.5%) (P < 0.001). Multivariate analysis showed that the risk for Q fever antibody positivity increased with age and was higher among persons who were foreign-born, male, and living in poverty. These findings indicate that the national seroprevalence of Q fever in the United States is higher than expected on the basis of case numbers reported to the Centers for Disease Control and Prevention from state health departments. Potential differences in risk for exposure by race/ethnicity warrant further study. PMID- 19815889 TI - The microscopic agglutination test (MAT) is an unreliable predictor of infecting Leptospira serovar in Thailand. AB - A prospective study in Thailand identified 106 patients with culture-proven leptospirosis. The accuracy of the microscopic agglutination test (MAT) in predicting the infecting serovar was evaluated in 78/106 (74%) patients with a diagnostic titer. MAT correctly determined the infecting serovar in 26 cases (33%), indicating that this assay is a poor predictor of infecting serovar in our setting. PMID- 19815890 TI - Clinical factors predictive of encephalitis caused by Angiostrongylus cantonensis. AB - Angiostrongylus cantonensis is mainly caused eosinophilic meningitis in humans, whereas a minority of patients develop encephalitic angiostrongyliasis (EA). EA is an extremely fatal condition, and the clinical factors predictive of EA have never been reported. A comparison study was conducted in a hospital situated in an endemic area of Thailand. We enrolled 14 and 80 angiostrongyliasis patients who developed encephalitis and meningitis, respectively. Logistic regression analysis was used to assess the clinical variables predictive of encephalitis. Age (adjusted odds ratio [OR], 1.22; 95% confidence interval [CI], 1.05-1.42), duration of headache (adjusted OR, 1.26; 95% CI, 1.03-1.55), and fever > 38.0 degrees C (adjusted OR, 37.05; 95% CI, 1.59-862.35) were identified as statistically significant factors for EA prediction. Elderly patients with angiostrongyliasis experiencing fever and prolonged headaches were at the highest risk of developing EA. PMID- 19815891 TI - Macrofilaricidal activity and amelioration of lymphatic pathology in bancroftian filariasis after 3 weeks of doxycycline followed by single-dose diethylcarbamazine. AB - In a placebo controlled trial, the effects of 21- and 10-day doxycycline treatments (200 mg/day) followed by single dose diethylcarbamazine (administered 4 months post treatment) on depletion of Wolbachia endobacteria from Wuchereria bancrofti, filaricidal activity, and amerlioration of scrotal lymph vessel dilation were studied in 57 men from Orissa, India. The 21-day doxycycline course reduced Wolbachia in W. bancrofti by 94% before diethylcarbamazine administration. After 12 months, all patients with this treatment were amicrofilaremic and different from the 10-day doxycycline (42.9%) and placebo (37.5%) groups, and significantly fewer were positive for scrotal worm nests (6.7%) compared with 10-day doxycycline (60%) and placebo (66.7%). Average scrotal lymph vessel diameters were reduced from 0.7 cm pre-treatment to 0.02 cm in patients after 21 days of treatment, while no significant changes were seen in the other groups. This latter feature confirms the beneficial effects of doxycycline on lymphatic dilation and thus adds to the existing evidence that doxycycline, in addition to being macrofilaricidal, may be used to prevent or reverse lymphatic pathology. PMID- 19815892 TI - Intra-cystic drug concentration of albendazole sulphoxide in patients with Echinococcus granulosus cysts. AB - Albendazole therapy--alone or in combination with surgery--remains the standard of care for human echinococcosis depending on the stage of disease. However, only limited data are available on target site concentrations in liver cysts and data for non-liver cysts are lacking. We report on intra-cystic concentrations of the biologically active metabolite albendazole sulphoxide in non-liver cysts indicating a relative intra-cystic drug concentration of 48-156% compared with plasma levels. These data are evidence for a satisfactory penetration of albendazole sulphoxide into Echinococcus cysts localized in other organs than the liver and underline the usefulness of albendazole therapy for this indication. PMID- 19815893 TI - Spatial variation of Yersinia pestis from Yunnan Province of China. AB - Yunnan Province of China is considered the site of origin for modern plague. We analyzed the genotypes of eight Yersinia pestis strains isolated from three counties in Yunnan Province by pulse field gel electrophoresis (PFGE). PFGE showed that strains isolated from the same site were identical regardless of hosts or year of isolation. However, Y. pestis strains isolated from geographically distinct loci were genetically divergent. Whole genome sequences of two strains from two foci in Yunnan showed that the genetic variation of Y. pestis strains was caused by genome rearrangement. We concluded that Y. pestis strains in each epidemic focus in Yunnan were a clonal population and selected by host environments. The genomic variability of the Y. pestis strains from different foci were caused by genome rearrangement, which may provide a positive selective advantage for Y. pestis to adapt to its host environments. PMID- 19815894 TI - Flea diversity and infestation prevalence on rodents in a plague-endemic region of Uganda. AB - In Uganda, the West Nile region is the primary epidemiologic focus for plague. The aims of this study were to 1) describe flea-host associations within a plague endemic region of Uganda, 2) compare flea loads between villages with or without a history of reported human plague cases and between sampling periods, and 3) determine vector loads on small mammal hosts in domestic, peridomestic, and sylvatic settings. We report that the roof rat, Rattus rattus, is the most common rodent collected in human dwellings in each of the 10 villages within the two districts sampled. These rats were commonly infested with efficient Y. pestis vectors, Xenopsylla cheopis and X. brasiliensis in Arua and Nebbi districts, respectively. In peridomestic and sylvatic areas in both districts, the Nile rat, Arvicanthus niloticus, was the most abundant rodent and hosted the highest diversity of flea species. When significant temporal differences in flea loads were detected, they were typically lower during the dry month of January. We did not detect any significant differences in small mammal abundance or flea loads between villages with our without a history of human plague, indicating that conditions during inter-epizootic periods are similar between these areas. Future studies are needed to determine whether flea abundance or species composition changes during epizootics when humans are most at risk of exposure. PMID- 19815895 TI - Syndromic approach to treatment of snake bite in Sri Lanka based on results of a prospective national hospital-based survey of patients envenomed by identified snakes. AB - Of 860 snakes brought to 10 hospitals in Sri Lanka with the patients they had bitten, 762 (89%) were venomous. Russell's vipers (Daboia russelii) and hump nosed pit vipers (Hypnale hypnale) were the most numerous and H. hypnale was the most widely distributed. Fifty-one (6%) were misidentified by hospital staff, causing inappropriate antivenom treatment of 13 patients. Distinctive clinical syndromes were identified to aid species diagnosis in most cases of snake bite in Sri Lanka where the biting species is unknown. Diagnostic sensitivities and specificities of these syndromes for envenoming were 78% and 96% by Naja naja, 66% and 100% by Bungarus caeruleus, 14% and 100% by Daboia russelii, and 10% and 97% by Hypnale hypnale, respectively. Although only polyspecific antivenoms are used in Sri Lanka, species diagnosis remains important to anticipate life threatening complications such as local necrosis, hemorrhage and renal and respiratory failure and to identify likely victims of envenoming by H. hypnale who will not benefit from existing antivenoms. The technique of hospital-based collection, labeling and preservation of dead snakes brought by bitten patients is recommended for rapid assessment of a country's medically-important herpetofauna. PMID- 19815896 TI - Proteinuria in women with preeclampsia: understanding the dialogue between 2 neighbors. PMID- 19815897 TI - Deathbed phenomena: its role in peaceful death and terminal restlessness. AB - Dying patients and their caregivers frequently experience that which is known as deathbed phenomena, that is, visions of past deceased relatives or friends, religious figures, and a visionary language pertaining to travel. Collective research supports mounting evidence that deathbed visions typically yield peaceful deaths. Yet within the literature, numerous hospice patients experience the symptoms of terminal restlessness and frequently succumb to anguished deaths. Why are some patients and caregivers guided by peaceful deathbed phenomena and others are not? Does a relationship exist between the lack of deathbed phenomena and the onset of terminal restlessness in dying patients? This clinical paper intends to answer these questions and might elucidate the factors that contribute to a dying patient's death ending as either a peaceful event or the one affected by terminal restlessness. This knowledge gained could lessen the occurrence of anguished deaths and perhaps change our way of viewing dying. PMID- 19815898 TI - Documentation of resuscitation status in an ambulatory palliative care population: results of a prospective observational study from a tertiary cancer care centre in Pakistan. AB - BACKGROUND: It was observed in our hospital that a lot of patients with advanced progressive disease were being seen in the palliative care clinics without documentation of their resuscitation status. AIM: To assess the documentation of resuscitation in patients referred to palliative care clinic. METHODS: Retrospective review of medical records of patients referred to the palliative care clinic was done looking for evidence of documentation of code status. RESULTS: Only 77 of the 316 patients seen in clinic during this period had any documentation of code status. More than half of these had been referred by the internists who had documented the code status as well. CONCLUSIONS: As a result of the study, we have introduced changes aimed at ensuring better documentation of resuscitation status. PMID- 19815899 TI - Medical students' views and ideas about palliative care communication training. AB - This study focused on the undergraduate medical student to identify views and ideas held toward palliative care communication training, pedagogical approaches to this training, and its perceived effectiveness and use in the medical field. Two focus groups consisting of fourth-year medical students were conducted, and their responses were analyzed using grounded theory categorization. Results indicated that students: (a) prefer to learn nonverbal communication techniques, (b) believe that natural ability and experience outweigh communication curriculum, (c) view the skill of breaking bad news as largely dependent on knowledge and expertise, and (d) prefer curriculum on palliative care and hospice to consist of information (eg, advance directives) rather than communication skills. Implications for these interpretive themes are discussed as well as future research and practice. PMID- 19815900 TI - Integration of diagnostic and communication technologies. AB - Three key areas in diagnostics will drive the convergence of diagnostic and communication technologies: point-of-care testing, micro-electromechanical systems and biomarker discovery. In addition, the communications revolution means that increasing numbers of people will be able to send data from their home to their doctor using the Internet. Also, the widespread availability of broadband opens up the possibly of realtime videoconferencing with clinicians. It is already possible for patients at home to monitor simple variables, such as heart rate and blood pressure, and send their results using communication technologies to their doctors, who can promptly review the information to diagnose problems. As diagnostic and communication technologies converge, it will be feasible for patients to transmit more complex health-care data periodically to their doctor, who will be able to identify problems early on and thus modify disease management to prevent exacerbations of patients' medical conditions. This will allow improved patient care in a wide range of health-care situations, from acute medical conditions to chronic disease. PMID- 19815901 TI - Technologically-assisted behaviour change: a systematic review of studies of novel technologies for the management of chronic illness. AB - A systematic review was conducted to investigate the use of technology in achieving behaviour change in chronic illness. The areas reviewed were: (1) methods employed to adapt traditional therapy from a face-to-face medium to a computer-assisted platform; (2) targets of behaviour change; and (3) level of human (e.g. therapist) involvement. The initial literature search produced 2032 articles. A total of 45 articles reporting 33 separate interventions met the inclusion/exclusion criteria and were reviewed in detail. The majority of interventions reported a theoretical basis, with many arising from a cognitive behavioural framework. There was a wide range of therapy content. Therapist involvement was reported in 73% of the interventions. A common problem was high participant attrition, which may have been related to reduced levels of human interaction. Instigating successful behaviour change through technological interventions poses many difficulties. However, there are potential benefits of delivering therapy in this way. For people with long-term health conditions, technological self-management systems could provide a practical method of understanding and monitoring their condition, as well as therapeutic guidance to alter maladaptive behaviour. PMID- 19815902 TI - Application of camera phones in telehaematology. AB - We investigated the use of camera phones for telehaematology. First, the minimum requirements for the camera phones to be used in telehaematology were investigated. A single image containing white cells, red cells and platelets was sent from a camera phone to 33 different camera phones. Nine of the camera phones were found to be unsuitable for telehaematology due to low display resolution or no zoom function of the image. Then we examined the agreement between a haematologist using a suitable camera phone for remote diagnosis and the blood film report made in the usual way. Blood samples were collected from nine patients who had conditions in which diagnostically important morphological abnormalities occurred. In seven of the nine cases, the telehaematology responses were similar to the documented blood film reports. We conclude that telehaematology using camera phones offers a quick and potentially valuable method of support for the diagnostic haematology laboratory. PMID- 19815903 TI - A comparison of in-person home care, home care with telephone contact and home care with telemonitoring for disease management. AB - We compared the effects of evidence-based disease management guidelines delivered to patients with heart failure and diabetes using three different modalities: in person visits alone (Control), in-person visits and a telephone intervention (Telephone), and in-person visits and telemonitoring (Telemonitoring). Patients were randomized to the three groups. There were 112 patients in the Control group, 93 in the Telephone group and 98 in the Telemonitoring group. During the first 60 days, 10% of the Control group were rehospitalized, 17% of the Telephone group and 16% of the Telemonitoring group. Having heart failure and receiving more in-person visits were significantly related to readmission and time to readmission. However, after adjusting for diagnosis and visits, the differences between the three groups were non-significant. There was a trend for increased risk of readmission for the Telephone group compared to Control alone (P = 0.07, risk ratio 2.2, 95% CI: 0.9 to 5.2) and for readmission sooner (P = 0.02, risk ratio 2.3, 95% CI: 1.2 to 4.6). Patient rehospitalization and emergency department visit rates were lower than the national average, making it difficult to detect a difference between groups. Previous rehospitalization was a consistent predictor of those who were rehospitalized, suggesting that it may be a useful indicator for identifying patients likely to need additional attention. PMID- 19815904 TI - Interactive emergency communication involving persons in crisis. AB - We studied the dialogue between telephone operators at medical emergency communication centres in Norway and parents of children later diagnosed with sudden infant death syndrome. The aim was to understand how the parents experienced the communication with the telephone operators. The qualitative method involved semi-structured interviews. We interviewed six respondents from urban areas and five from rural areas. An important finding was that all the parents were satisfied with the resuscitation instructions they received. It was also perceived as important that the emergency operators expressed empathy and care. We believe that it is not merely the quality of the resuscitation attempts that the operators' efforts should be measured against. It is also important that the operators provide good explanations and express emotional support. Our findings indicate that this will be enormously appreciated, even if callers do not feel that they are capable of performing optimum resuscitation. PMID- 19815905 TI - Telehealth in clinical supervision: a comparison of supervision formats. AB - Nine counselling psychology students were enrolled in a 12-week pilot practicum (i.e. a work placement) for either one hour of course credit (six students) or three hours (three students). Group supervision was provided both in-person and by videoconferencing. Each trainee completed a measure evaluating their satisfaction with supervision (Supervisory Satisfaction Questionnaire, SSQ) and the supervisory relationship (Supervisory Working Alliance Inventory-Trainee Version, SWAI-T). The student's self-efficacy was also tracked during the semester (Counselling Self-Estimate Inventory, COSE). Trainees rated their satisfaction with videoconferencing similarly to the in-person format. The supervisory relationship also did not appear to be affected by the videoconferencing format. The COSE scores indicated that the students increased in counsellor self-efficacy by the end of the semester. Trainees reported that their supervisory needs were met and believed that videoconferencing was a viable format for supervision, although such a format still needed to be augmented by in person contact. Providing better access to supervision and professional support using technology is one step towards improving health care in rural areas. PMID- 19815906 TI - Feasibility of telemonitoring for active surveillance of influenza vaccine safety in the primary care setting in The Netherlands. AB - We examined the feasibility of a commercial home telemonitoring system for monitoring adverse events related to vaccination and influenza-like illness (ILI) signs and outcomes in the primary care setting in The Netherlands. A prospective cohort of people eligible for influenza vaccination was monitored daily between mid-October 2007 and mid-March 2008. Adults from five primary care centres were invited to participate. A total of 245 people participated (response rate 75%). Their mean age was 61 years (SD = 15), 50% were female and 60% had a chronic disease. Most (73%) had no problems with installation of the system and 67% finished all sets of monitoring dialogues. The reported incidence of adverse events in the first week after vaccination was 8-38%. The reported incidence rates of ILI symptoms varied and were higher than reference data. A total of 39% of individuals consulted their general practitioner, 7% the hospital emergency department, 6% were hospitalized and 27% used medication. Of those in paid work, one-third reported absence of work due to ILI. Home telemonitoring appears to be feasible for monitoring vaccine adverse events and ILI symptoms and outcomes. PMID- 19815908 TI - A Brazilian educational experiment: teleradiology on web TV. AB - Since 2004, educational videoconferences have been held in Brazil for paediatric radiologists in training. The RUTE network has been used, a high-speed national research and education network. Twelve videoconferences were recorded by the Health Channel and transformed into TV programmes, both for conventional broadcast and for access via the Internet. Between October 2007 and December 2009 the Health Channel website registered 2378 hits. Our experience suggests that for successful recording of multipoint videoconferences, four areas are important: (1) a pre-planned script is required, for both physicians and film-makers; (2) particular care is necessary when editing the audiovisual material; (3) the audio and video equipment requires careful adjustment to preserve clinical discussions and the quality of radiology images; (4) to produce a product suitable for both TV sets and computer devices, the master tape needs to be encoded in low resolution digital video formats for Internet media (wmv and rm format for streaming, and compressed zip files for downloading) and MPEG format for DVDs. PMID- 19815907 TI - Pandemic influenza planning by videoconference. AB - Collaboration between nations and sectors is crucial to improve regional preparedness against pandemic influenza. In 2008, a Virtual Symposium was organized in the Asia-Pacific region by the Asia-Pacific Economic Cooperation Emerging Infections Network (APEC EINet) to discuss pandemic preparedness. The multipoint videoconference lasted approximately 4.5 hours and was attended by 16 APEC members who shared best practices in public-private partnerships for pandemic influenza preparedness planning. Twelve of the 16 APEC members who participated responded to a post-event survey. The overall experience of the event was rated highly. Partnering public health, technology and business communities to discuss best practices in preparedness using videoconferencing may be an effective way to improve regional preparedness. Utilization of videoconferencing on a routine basis should be considered to improve preparedness among APEC members and enhance its usability during a pandemic. PMID- 19815909 TI - Review of drug treatment for female sexual dysfunction. AB - Female sexual dysfunction (FSD) incorporates various sexual disorders including hypoactive sexual desire disorder, sexual arousal disorder, orgasmic and sexual pain disorders. Although many strategies have been formulated for the treatment of male sexual problems, FSD remains an area that warrants further research and trial studies to identify the most efficacious treatment options. Research has highlighted numerous pharmacological interventions that have been trialled and found to exhibit positive effects. These include hormonal therapies, prostaglandins, dopaminergic agonists, phosphodiesterase type-5 (PDE-5) inhibitors and melanocortin agonists. PMID- 19815910 TI - A review of cervical cytology in genitourinary medicine clinics in England between 1997 and 2008. AB - The current cervical screening guidelines lay the duty of care of women under 25 years of age on genitourinary (GU) medicine clinics. To date there has not been an extensive review of cervical cytology results sourced from GU medicine departments. We reviewed the annual reports of the cervical screening programme, with their extensive and comparative data, including records of cervical cytology from GU medicine. There is a clear and progressive trend of rising percentages of 'severe dyskaryosis' in laboratory reported GU medicine cervical smears. The trend and observations indicate that GU medicine physicians should take young women's concern on cervical cancer seriously. The programme of opportunistic cervical screening in GU medicine should not be discouraged. PMID- 19815911 TI - Concurrent sexual partnerships among individuals in HIV sero-discordant heterosexual couples. AB - The objective of the study was to assess the level at which individuals in HIV discordant couples engage in concurrent sexual partnerships and factors associated with this risk of transmitting HIV. By using data from a group of HIV discordant heterosexual couples (n = 145), we examined the frequency of concurrent sexual partners and factors associated with such partnerships. The prevalence of concurrent partnerships with heterosexual partners was 16%. Fewer than half of individuals with concurrent partnerships reported that their main study partner knew about these relationships. Of individuals involved in concurrent partnerships, 30% reported inconsistent condom use in these relationships. Unmarried individuals in new HIV sero-discordant relationships were particularly at risk for concurrent partnerships. In conclusion, more frequent HIV testing may complement increased attention to communication, disclosure and condom use in this population that is especially vulnerable to acquiring and transmitting HIV infection. PMID- 19815912 TI - Cardiovascular risk factors and ultrasound evaluation of carotid atherosclerosis in patients with HIV-1 infection. AB - A cross-sectional study was performed to evaluate classical risk factors for cardiovascular diseases and subclinical atherosclerosis by carotid ultrasonography in HIV-positive subjects, naive or treated with antiretroviral agents. A total of 66 patients were enrolled into the study: 21 subjects were naive to all antiretroviral agents (group A) and 45 patients were treated with antiretroviral therapy for >or=36 months (group B). The prevalence of carotid plaques was significantly higher in group B than in group A (44.7% versus 0%; P = 0.014). In group B, patients with high 10-year risk of coronary heart disease showed a significantly higher intima-media thickness and prevalence of carotid lesions than those with low risk. Moreover, carotid lesions were structurally comparable to classical atherosclerotique plaques observed in the general population, with iso-hyperechonegic aspects and irregular surfaces. The prevalence of carotid atherosclerosis in experienced patients is higher than in those naive to highly active antiretroviral therapy and seems mostly associated with a longer duration of HIV infection, more severe lipid metabolism alterations, presence of lipodystrophy syndrome and a more elevated 10-year risk of cardiovascular diseases. PMID- 19815914 TI - The psychological impact caused by genital warts: has the Department of Health's choice of vaccination missed the opportunity to prevent such morbidity? AB - This study evaluated the psychological impact caused by genital warts and whether this altered over time. Eighty-four patients with genital warts and a group of 28 appropriately matched volunteers from the local general population were recruited. The patients were divided into three groups of 28 according to the Health Protection Agency classification codes for genital warts. A comprehensive health questionnaire was completed by all participants. Patients with first attack genital warts experienced significantly more distress, anxiety and depression and were less satisfied with their lives than all the other groups. They also reported less satisfaction with the practical support they received. The recurrent episode genital warts group experienced significantly high levels of emotional distress and the re-registered group reported significantly less emotional and practical support according to their needs. While the first attack genital warts group experienced the most psychological effects, it is clear that those effects did not disappear as an individual came to terms with their diagnosis. The choice of vaccine to prevent cervical cancer will not prevent clinical warts occurring and therefore will not help prevent the psychological morbidity associated with this condition. PMID- 19815915 TI - What service users want: a new clinic results service. Can we satisfy both patients' needs and wants? AB - Many clinics still have a 'no news is good news' (NNIGN) policy for clinic results. We asked our service users their preferences for obtaining results. We also designed a new clinic results service. Patients were invited to complete a service evaluation questionnaire. They were given nine options for obtaining results and asked to rate acceptability on a 1-9 scale (unacceptable to acceptable). Completed questionnaires were compared with the actual method they chose to obtain results for that visit. In all, 1000 questionnaires (561 females, 439 males) were completed. Average acceptability score was highest for mobile phone (7.7), followed by text (5.8) and letter (5.7). NNIGN (3.7) and returning for results (3.6) received the least popular ratings. When compared with the actual method chosen, mobile phone was the most popular (62%) followed by letter (17%). Only 10% of patients found NNIGN acceptable. Following the evaluation we purchased texting software and redesigned our results service. There are now four options for receiving results - texting, letter, patient telephones us and NNIGN. We believe we have been able to satisfy both patients' needs and their wishes by redesigning our service around their views. PMID- 19815913 TI - A randomized double-blind comparison of moxifloxacin and doxycycline/metronidazole/ciprofloxacin in the treatment of acute, uncomplicated pelvic inflammatory disease. AB - This multicentre, double-blind study was undertaken to demonstrate non inferiority of once-daily oral moxifloxacin compared with combination therapy in the management of acute, uncomplicated pelvic inflammatory disease (PID). Women aged >or=18 years with PID were randomized to receive moxifloxacin (400 mg once daily) for 14 days or comparator treatment (doxycycline [100 mg twice daily] plus metronidazole [400 mg three times daily] for 14 days, plus one single 500-mg ciprofloxacin dose). Of the 434 valid per protocol (PP) patients, the overall clinical success rates at 2-14 days post-therapy were 96.6% (moxifloxacin) and 98.0% (comparator); moxifloxacin was non-inferior to the comparator regimen both in the PP (95% confidence interval [CI]: -4.5, 1.6) and intent-to-treat (95% CI: 5.8, 6.9) populations. Clinical success rates at 21-35 days post-therapy were 93.8% (166/177; data missing for 47 patients) for moxifloxacin and 91.3% (147/161; data missing for 37 patients) for the comparator. Bacteriological success rates at 2-14 days post-therapy were 92.5% (moxifloxacin) and 88.2% (comparator). Once-daily dosing and proven efficacy suggest that moxifloxacin may be of value in acute, uncomplicated PID. PMID- 19815916 TI - Factors associated with unprotected anal intercourse between HIV-positive men and regular male partners in a Sydney cohort. AB - Blood plasma HIV-RNA load (BPVL) is the strongest predictor of HIV-1 transmission during sex. Unprotected anal intercourse (UAI) is the highest risk activity for transmission among men who have sex with men (MSM). Awareness of BPVL may influence rates of UAI. We assessed whether optimism towards antiretroviral therapy (ART) and/or biomedical factors influenced sexual activities with regular partners. Questionnaires were administered to 109 HIV-positive MSM participating in a cross-sectional study of BPVL and seminal viral load. The survey assessed HIV transmission beliefs and sexual practices with regular male partners in the past three months. Sixty-nine of 109 (63.3%) had been in a regular relationship and 42 reported having had anal sex. Unprotected receptive anal intercourse without ejaculation (URAI - e) was associated with awareness that their most recent BPVL was detectable (>50 RNA copies/mL) and not taking ART. Receptive UAI with ejaculation (URAI + e) was associated with not taking ART, having a sexually transmissible infection and having an HIV-positive partner; the latter was also associated with insertive UAI with ejaculation (UIAI + e). Treatment optimism was not associated with UAI. In this cohort, sexual practices were based more upon knowledge of biomedical factors rather than attitudes regarding transmission risks. PMID- 19815917 TI - Profile of direct walk-in and referred clients attending Integrated Counselling and Testing Centre. AB - The HIV pandemic has had a profound impact on the health and economic conditions of individuals, and people living with HIV/AIDS are faced with the task of maintaining optimal health status despite an increasing insult to their immune status. The aim of the present study was to study the profile of direct walk-in and referred patients attending the Integrated Counselling and Testing Centre (ICTC) of a tertiary care hospital, which may provide important clues to understanding the epidemiology of the disease in a particular region. The study included all the attendees of the ICTC referred from the hospital or direct walk in from January 2007 to December 2007. Three rapid HIV tests were used and the samples showing positive results in all the three tests were declared HIV positive. The results were analysed to correlate between HIV positivity, age, sex, route of transmission and direct walk-in/referred patients. A low proportion of ICTC attendees (27%) in our study were direct walk-ins. As regards HIV positivity, 312 (8.3%) out of 2440 males and 164 (4.3%) out of 1315 females were HIV positive. Among the referred HIV-positive patients 162 were males and 62 females, whereas among the direct walk-in HIV-positive patients, 150 were males and 102 females. Integrated counselling and testing is now seen as a key entry point for HIV prevention. In addition to scaling up ICTC services, it is also important to raise awareness by aggressive health education programmes and integration of ICTC into various community organizations. PMID- 19815918 TI - What do patients want most from sexual health services? AB - Changes to sexual health clinics have recently been driven by government waiting time targets. However, patients' needs and expectations should be equally important when making service alterations. This study sought to determine what patients valued most when attending an integrated sexual health clinic. During February 2009, 252 patients attending a sexual health clinic in outer London completed an anonymous questionnaire. The questionnaire asked for their views on different aspects of sexual health care. The results showed that the most highly valued aspects of care were confidentiality (18.47% of points allocated) followed by speed of service (13.1%) and rapid test results (12.12%). These aspects were more important than being seen within 48 hours (7.78%), technical expertise (6.26%) or other patient-centred aspects of care. These findings, which represent patients' views, have important implications for service design. PMID- 19815919 TI - Bronzino's 'Allegory of Venus and Cupid': an exemplary image for contemporary sexual health promotion? AB - Recent sexual health promotion strategies have veered between a negative emphasis on the deleterious consequences of sexually transmitted infections, and a more positive, eroticized approach to safer sex. The differences in approach are starkly reflected in the images chosen to illustrate them. We note that there are problems with both approaches. The main purpose of this review is to demonstrate how this dichotomy was transcended by the sixteenth century Florentine Mannerist painter, Agnolo Bronzino, in his allegory on syphilis. PMID- 19815920 TI - Successful treatment of a large Buschke-Lowenstein tumour with chemo radiotherapy. AB - We report the case of a patient with a large Buschke-Lowenstein tumour which had previously recurred following local excision. A preferred treatment modality for this rare variant of human papillomavirus has not been clearly defined. Treatment with chemo-radiotherapy in this case resulted in complete resolution of the disease without the need for further surgical intervention. PMID- 19815921 TI - Preliminary HIV test result disclosure: lessons we can learn. AB - Result disclosure following routine HIV testing can be challenging. We present an interesting real life scenario regarding a young heterosexual Caucasian male patient, who was diagnosed as having a positive HIV antibody result following a routine HIV screening test, his confirmatory test result was HIV negative. This article covers important lessons learnt from the case. PMID- 19815922 TI - Obstructive endo-bronchial pseudotumour due to herpes simplex type 2 infection in an HIV-infected man. AB - We report a 31-year-old man with an obstructive bronchial lesion due to herpes simplex type 2 infection, who responded promptly to endoscopic resection and oral treatment with acyclovir. Exophytic lesions of the respiratory tract are rare, potentially life-threatening, but readily treated complication of herpes simplex virus infection in HIV-infected individuals. PMID- 19815923 TI - Response to Crum-Cianflone et al., syphilitic hepatitis among HIV-infected patients. PMID- 19815924 TI - Oral-anal intercourse and sexual transmission of Enterobius vermicularis; do we need to screen for other intestinal parasites? PMID- 19815925 TI - Preference for 50% reinforcement over 75% reinforcement by pigeons. AB - When pigeons are given a choice between an initial-link alternative that results in either a terminal-link stimulus correlated with 100% reinforcement or a stimulus correlated with 0% reinforcement (overall 50% reinforcement) and another initial-link alternative that always results in a terminal-link stimulus correlated with 100% reinforcement, some pigeons show a preference for the initial-link alternative correlated with 50% reinforcement. Using this procedure, in Experiment 1, we found a relatively modest preference for 100% over 50% reinforcement. In Experiment 2, we decreased the reinforcement density for the second initial-link alternative to 75% and found a significant preference for the 50% reinforcement initial-link alternative. It may be that this "maladaptive" behavior results from a positive contrast between the expectation of reinforcement correlated with the 50% reinforcement initial-link alternative and the terminal-link stimulus correlated with 100% reinforcement. But apparently, the complementary negative contrast does not develop between the expectation of reinforcement correlated with the 50% reinforcement initial-link alternative and the terminal-link stimulus correlated with 0% reinforcement that often follow. Such paradoxical choice may account for certain human appetitive risk-taking behavior (e.g., gambling) as well. PMID- 19815926 TI - Animal memory: the contribution of generalization decrement to delayed conditional discrimination retention functions. AB - Delayed conditional discriminations in which a sample indicates which comparison stimulus is correct have typically been used in working memory research with animals. Following acquisition with no (0-sec) delay between the offset of the sample and the onset of the comparison stimuli, delays of variable duration are introduced. The resulting retention functions are taken as a measure of memory. We suggest that, in addition to memory loss due to the delay, the comparison of matching accuracy at the 0-sec training delay with relatively novel test delays may produce a generalization decrement that varies as a function of increasing delay. We tested this hypothesis by training pigeons with a mixed delay procedure from the start and found that the retention functions for these pigeons were significantly shallower than those for a control group trained with 0-sec delays and tested with longer delays, and, although reduced in magnitude, the differences persisted for as many as 15 sessions. We propose that a measure of animals' working memory can be obtained uninfluenced by a generalization decrement if they have received comparable training with all of the delays that are tested. PMID- 19815927 TI - A learned flavor preference persists despite the extinction of conditioned hedonic reactions to the cue flavors. AB - Learned flavor preferences can be strikingly persistent in the face of behavioral extinction. Harris, Shand, Carroll, and Westbrook (2004) suggested that this persistence may be due to flavor preference conditioning's producing a long lasting change in the hedonic response to the conditioned stimulus (CS+) flavor. In the present study, the CS+ flavor was presented in simultaneous compound with 16% sucrose, whereas the CS- flavor was presented with 2% sucrose. During subsequent two- and one-bottle tests, the CS+ and CS- flavors were presented in 2% sucrose. Hedonic reactions during training and test were assessed using an analysis of the microstructure of licking behavior. Conditioning resulted in greater consumption of the CS+ than of the CS- that did not extinguish over repeated two- and one-bottle tests. The mean lick cluster size was higher for the CS+ than for the CS- only on the first cycle of tests. Since lick cluster size can be used as an index of stimulus palatability, the present results indicate that although the hedonic reaction to the CSs did change, this was not maintained across repeated tests. Thus, changes in the hedonic response to the conditioned flavors cannot explain the resistance to the extinction of learned flavor preferences. PMID- 19815928 TI - Effects of pretraining on acquisition of novel configural discriminations in human predictive learning. AB - In two experiments, participants acquired one of two target configural discriminations (a biconditional or negative patterning discrimination) in a predictive learning task. In Experiment 1, participants were pretrained with either a configural or an elemental discrimination; in Experiment 2, they were pretrained with a configural discrimination, an elemental discrimination, or a control discrimination that was not expected to bias them toward elemental or configural processing. In both experiments, acquisition of the target configural discriminations was faster after configural pretraining than after elemental pretraining. In addition, the negative patterning discrimination was acquired faster than the biconditional discrimination. Finally, the results of Experiment 2 were more consistent with the notion that elemental pretraining hindered acquisition of the target discriminations than with the notion that configural pretraining enhanced their acquisition. Implications of these findings are discussed. PMID- 19815929 TI - Learning-related shifts in generalization gradients for complex sounds. AB - Learning to discriminate stimuli can alter how one distinguishes related stimuli. For instance, training an individual to differentiate between two stimuli along a single dimension can alter how that individual generalizes learned responses. In this study, we examined the persistence of shifts in generalization gradients after training with sounds. University students were trained to differentiate two sounds that varied along a complex acoustic dimension. The students were subsequently tested on their ability to recognize a sound that they had experienced during training when it was presented among several novel sounds varying along this same dimension. Peak shift was observed in Experiment 1, in which generalization tests immediately followed training, and in Experiment 2, in which the tests were delayed by 24 h. These findings further support the universality of generalization processes across species, modalities, and levels of stimulus complexity. They also raise new questions about the mechanisms underlying learning-related shifts in generalization gradients. The sound stimuli from this study are available as .wav files from http://lb.psychonomic journals.org/content/supplemental. PMID- 19815930 TI - Generalization of causal efficacy judgments after evaluative learning. AB - In three experiments, we examined the effect of response-outcome relations on human ratings of causal efficacy and demonstrated that such efficacy ratings transfer to novel situations through derived stimulus relations. Causal efficacy ratings were higher, and probability of an outcome given a response was lower, for a differential reinforcement of high rate schedule than for either a differential reinforcement of low rate schedule (Experiment 1) or a variable interval schedule (Experiment 2). In Experiment 3, we employed schedules that were equated for outcome probability and noted that ratings of causal efficacy and the rate of response were higher on a variable ratio than on a variable interval schedule. For participants in all three experiments, causal efficacy ratings transferred to the stimulus present during each schedule and generalized to novel stimuli through derived relations. The results corroborate the view that schedules are a determinant of both response rates and causal efficacy ratings. In addition, the novel demonstration of a mechanism of generalization of these ratings via derived relations has clinical implications. PMID- 19815931 TI - Comparison of auditory and visual conditioning stimuli in delay eyeblink conditioning in healthy young adults. AB - Classical eyeblink conditioning (EBC) has been widely used to probe cerebellar function in humans and nonhuman mammals. Although the neural pathways governing behavior in this task are well understood and fairly discrete, it remains unclear in the human literature how conditioned stimuli (CSs) of different modalities (e.g., visual and auditory) influence the exhibition of conditioned responses (CRs). In the present study, therefore, CRs to a visual CS and an auditory CS were examined with the single-cue delay EBC procedure. An initial experiment (N = 61) was conducted to identify visual and auditory stimuli that had equal perceived intensities. Using these perceptually equivalent stimuli, a second group of 25 subjects completed auditory and visual EBC procedures in two testing sessions 5-8 days apart. Whereas the acquisition of CRs was similar between the CS modality conditions, the timing of the CRs differed such that earlier CR onset and peak latencies were associated with the visual CS. In addition, CR timing improved across testing sessions, as indicated by the later CR peak latencies exhibited during the second testing session, as compared with the first. PMID- 19815932 TI - Behavioral momentum and relapse of extinguished operant responding. AB - Previous experiments on behavioral momentum have shown that relative resistance to extinction of operant behavior in the presence of a stimulus depends on the rate of reinforcement associated with that stimulus, even if some of those reinforcers occur independently of the behavior. We present three experiments examining whether the rate of reinforcement in the presence of a stimulus similarly modulates the relative relapse of operant behavior produced by reinstatement, resurgence, and renewal paradigms. During baseline conditions, pigeons responded for food reinforcement on variable-interval 120-sec schedules in alternating periods of exposure to two stimuli arranged by a multiple schedule. Additional response-independent food presentations were also delivered in the presence of one of the multiple-schedule stimuli. Consistent with previous research, baseline response rates were lower in the presence of the stimulus with the added response-independent reinforcement, and relative resistance to extinction was greater in the presence of that stimulus. In addition, following extinction, the relative relapse of responding produced by reinstatement, resurgence, and renewal paradigms was greater in the presence of the stimulus associated with the higher rate of reinforcement. We suggest that a model of extinction from behavioral momentum theory may be useful for understanding these results. PMID- 19815933 TI - Counting absolute numbers of items, from 1 to 8, in pigeons. AB - Pigeons were trained in a forced choice task with four alternatives to categorize arrays consisting of 1, 3, 5, or 8 dots. Before the pigeons chose a comparison stimulus, they were required to peck each dot sequentially. A single peck to a dot, which was defined as an indicating response, changed the color of the dot so that it was differentiated from those that remained to be counted. The pigeons successfully learned to categorize the numerical arrays and then displayed transfer to novel arrays consisting of two, four, six, or seven dots, in a manner according to the order of 1 < 2 < 3 < 4 < 5 < 6 < 7 < 8. Subsequent tests revealed that the pigeons discriminated the stimuli by relying on the number of indicating responses. They also utilized multiple information (surface area, time, and other confounded events), but this was of minor significance, and after training, the pigeons were able to disregard these cues. PMID- 19815935 TI - Comments from a former Journal of Virology editor. PMID- 19815934 TI - BANK1 functional variants are associated with susceptibility to diffuse systemic sclerosis in Caucasians. AB - OBJECTIVE: To investigate the possible association of the BANK1 gene with genetic susceptibility to systemic sclerosis (SSc) and its subphenotypes. METHODS: A large multicentre case-control association study including 2380 patients with SSc and 3270 healthy controls from six independent case-control sets of Caucasian ancestry (American, Spanish, Dutch, German, Swedish and Italian) was conducted. Three putative functional BANK1 polymorphisms (rs17266594 T/C, rs10516487 G/A, rs3733197 G/A) were selected as genetic markers and genotyped by Taqman 5 allelic discrimination assay. RESULTS: A significant association of the rs10516487 G and rs17266594 T alleles with SSc susceptibility was observed (pooled OR=1.12, 95% CI 1.03 to 1.22; p=0.01 and pooled OR=1.14, 95% CI 1.05 to 1.25; p=0.003, respectively), whereas the rs3733197 genetic variant showed no statistically significant deviation. Stratification for cutaneous SSc phenotype showed that the BANK1 rs10516487 G, rs17266594 T and rs3733197 G alleles were strongly associated with susceptibility to diffuse SSc (dcSSc) (pooled OR=1.20, 95% CI 1.05 to 1.37, p=0.005; pooled OR=1.23, 95% CI 1.08 to 1.41, p=0.001; pooled OR=1.15, 95% CI 1.02 to 1.31, p=0.02, respectively). Similarly, stratification for specific SSc autoantibodies showed that the association of BANK1 rs10516487, rs17266594 and rs3733197 polymorphisms was restricted to the subgroup of patients carrying anti topoisomerase I antibodies (pooled OR=1.20, 95% CI 1.02 to 1.41, p=0.03; pooled OR=1.24, 95% CI 1.05 to 1.46, p=0.01; pooled OR=1.26, 95% CI 1.07 to 1.47, p=0.004, respectively). CONCLUSION: The results suggest that the BANK1 gene confers susceptibility to SSc in general, and specifically to the dcSSc and anti topoisomerase I antibody subsets. PMID- 19815936 TI - 'Ethnicity testing' before adoption: a help or hindrance? PMID- 19815937 TI - Comparison of interferon-gamma release assays and tuberculin skin test in predicting active tuberculosis (TB) in children in the UK: a paediatric TB network study. AB - BACKGROUND: The value of interferon-gamma release assays (IGRA) to diagnose active tuberculosis (TB) in children is not established, but these assays are being widely used for this purpose. The authors examined the sensitivity of commercially available IGRA to diagnose active TB in children in the UK compared with the tuberculin skin test (TST). METHODS: The authors established a paediatric tuberculosis network and conducted a retrospective analysis of data from children investigated for active TB at six large UK paediatric centres. All centres had used TST and at least one of the commercially available IGRA (T Spot.TB or Quantiferon-Gold in Tube) in the diagnostic work-up for active TB. Data were available from 333 children aged 2 months to 16 years. The authors measured the sensitivity of TST and IGRA in definite (culture confirmed) and probable TB in children, agreement between TST and either IGRA, and their combined sensitivity. RESULTS: Of 333 children, 49 fulfilled the criteria of definite TB, and 146 had probable TB. Within the definite cohort, TST had a sensitivity of 82%, Quantiferon-Gold in tube (QFT-IT) had a sensitivity of 78% and T-Spot.TB of 66%. Neither IGRA performed significantly better than a TST with a cut-off of 15 mm. Combining the results of TST and IGRA increased the sensitivity to 96% for TST plus T-Spot.TB and 91% for TST plus QFG-IT in the definite TB cohort. CONCLUSIONS: A negative IGRA does not exclude active TB disease, but a combination of TST and IGRA increases the sensitivity for identifying children with active TB. PMID- 19815938 TI - Additional value of two-channel amplitude integrated EEG recording in full-term infants with unilateral brain injury. AB - BACKGROUND: Amplitude integrated electroencephalography (aEEG) is a valuable tool for evaluating neonatal encephalopathy and identifying electrographic seizures. OBJECTIVE: To compare seizure activity and background pattern (BGP) between one channel and two-channel aEEG recordings in full-term neonates. METHODS: The two channel aEEG recordings (F3-P3; F4-P4) of 34 neonates with seizures were compared with single-channel recordings (P3-P4). RESULTS: All 34 infants with unilateral (n=14), diffuse (n=18) or without (n=2) brain injury had seizure patterns on one channel and two-channel recordings, with 18% more seizure patterns detected with two-channel recording. In 79% of infants with unilateral injury more seizures were noted on the ipsilateral side compared to the contralateral side. In 39% of the infants with diffuse brain damage more seizures were found with two-channel recordings. A sensitivity of 65% was found when using the automatic seizure detection algorithm. In 4/14 (29%) infants with unilateral injury a more severely affected BGP was seen on the ipsilateral side compared to the BGP on one-channel recording. In infants with diffuse injury differences in BGP pattern were seen in 6-17% of the infants depending on the system used for scoring. CONCLUSION: Although there were no major differences found between seizure detection with one channel or two-channel aEEG, in a subgroup of infants with a predominantly unilateral brain lesion, two-channel recording did provide additional information with identification of more seizure patterns on the affected side, sometimes also associated with a difference in BGP. To improve early diagnosis of unilateral lesions and improve seizure detection in these infants, routine use of two channel recordings is recommended. PMID- 19815939 TI - Pain in neonates during screening for retinopathy of prematurity using binocular indirect ophthalmoscopy and wide-field digital retinal imaging: a randomised comparison. AB - OBJECTIVE: To compare the pain experienced by premature infants undergoing wide field digital retinal imaging (WFDRI) and binocular indirect ophthalmoscopy (BIO) for retinopathy of prematurity (ROP) screening. METHODS: Infants were recruited at Edinburgh Royal Infirmary Neonatal Unit, Edinburgh, UK. Eyes were examined by WFDRI and BIO with eyelid speculum by two experienced paediatric ophthalmologists in random order. A pain score (Premature Infant Pain Profile (PIPP)) for WFDRI and BIO was generated. RESULTS: A total of 76 infants were recruited. The (mean, SD) PIPP score for WFDRI was 15.0, 2.1 and for BIO was 15.2, 2.4 (paired t test p=0.47). The authors observed that infants started crying with corresponding physiological changes as soon as the eyelid speculum was inserted and crying stopped on speculum removal. CONCLUSION: WFDRI and BIO with eyelid speculum are similarly painful for infants. The authors speculate that the eyelid speculum rather than the examination method may contribute most to the pain experienced. PMID- 19815940 TI - Cardiac rehabilitation: into the future. AB - Cardiac rehabilitation is increasingly recognised as an integral component of comprehensive cardiac care. The evidence supporting its effectiveness in reducing morbidity and mortality and improving quality of life is compelling. Yet, despite this recognition and exhortations that its implementation should be a key priority, most cardiac patients do not receive rehabilitation. Service provision varies markedly and many programmes are focused on select populations, often operate in an inflexible manner and fail to add potential value. Issues of suboptimal referral, enrolment and completion are poorly addressed and the potential for embracing novel methods and the latest technology are rarely exploited. This paper reviews the current status of cardiac rehabilitation and proposes ways to improve access and uptake and reduce inequity to ensure that those who are likely to benefit from this complex intervention do so. PMID- 19815941 TI - Role of NF-kappa B in the pathogenesis of diabetes and its associated complications. AB - The nuclear factor-kappaB (NF-kappaB) family is comprised of DNA-binding protein factors that are required for the transcription of most proinflammatory molecules, including adhesion molecules, enzymes, cytokines, and chemokines. Studies performed in a variety of cell and animal based experimental systems suggest that NF-kappaB activation is a key event early in the pathobiology of diabetes. The purpose of this review is to summarize the current knowledge about the molecular biology of NF-kappaB and to review evidence that implicates NF kappaB in the pathogenesis of diabetes and its associated complications. PMID- 19815942 TI - Serum cortisol concentration in patients with major depression after treatment with clomipramine. AB - Hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis and elevated cortisol (CORT) levels are characteristics of the pathophysiology of major depressive disorder. The aim of this study was to determine whether increased plasma CORT levels appear in patients with major depression and if effective antidepressant treatment by clomipramine (CLO) leads to regulation of CORT level. Plasma CORT levels were measured using high performance liquid chromatography (HPLC) methods in patients with major depression at time zero (before therapy) and after 3 h, 24 h, 4, 6 and 8 weeks of CLO administration. The study included 17 patients (12 women, 5 men; mean age 54.5 years, SD =12.3) and 21 healthy comparison subjects. The patients had a mean score on the 21-item Hamilton Depression Rating Scale (HDRS) of 26.8 (range 22-35). Eight of the patients with major depression recruited for the study showed a 46% increase in CORT concentration compared to the established standard. In 13 patients treated with CLO, serum CLO levels reached a therapeutic range. In recovered depressed patients, antidepressant treatment significantly reduced HDRS scores from the 6th week of treatment. A drop in plasma CORT levels in recovered depressed subjects occurred 0 to 6 weeks after CLO treatment (n = 5, p < 0.046). However, neither subject group exhibited any definitive markers of CORT secretion. In the population studied, patients had distinct profiles of HPA axis dysregulation. Finding a linear correlation between lower CORT secretion and therapeutic plasma CLO levels is the first aim of monitored therapy and may be important for understanding the pathophysiology of major depressive disorder. PMID- 19815943 TI - Prenatal stress decreases glycogen synthase kinase-3 phosphorylation in the rat frontal cortex. AB - It has been postulated that hyperactive glycogen synthase kinase-3 (GSK-3) is an important factor in the pathogenesis of depression, and that this enzyme also contributes to the mechanism of antidepressant drug action. In the present study, we investigated the effect of prenatal stress (an animal model of depression) and long-term treatment with antidepressant drugs on the concentration of GSK-3beta and its main regulating protein kinase B (PKB, Akt). The concentration of GSK 3beta, its inactive form (phospho-Ser9-GSK-3beta), and the amounts of active (phospho-Akt) and total Akt were determined in the hippocampus and frontal cortex in rats. In order to verify our animal model of depression, immobility time in the forced swim test (Porsolt test) was also determined.We found that prenatally stressed rats display a high level of immobility in the Porsolt test and chronic treatment with imipramine, fluoxetine, mirtazapine and tianeptine normalize this change. Western blot analysis demonstrated that GSK-3beta levels were significantly elevated in the frontal cortex, but not in the hippocampus, of prenatally stressed rats. The concentration of its non-active form (phospho-Ser9 GSK-3beta) was decreased only in the former brain structure. No changes were found in the amounts of active (phospho-Akt) and total Akt in both studied brain structures. Chronic treatment with antidepressant drugs diminished stress-induced alterations in GSK-3beta and phospho-GSK-3beta the frontal cortex, but had no effect on the concentration of these enzymes in the hippocampus. Moreover, levels of Akt and phospho-Akt in all experimental groups remained unchanged. Since our animal model of depression is connected with hyperactivity of the HPA axis, our results suggest that GSK-3beta is an important intracellular target for maladaptive glucocorticoid action on frontal cortex neurons and in antidepressant drug effects. Furthermore, the influence of stress and antidepressant drugs on GSK-3beta does not appear to impact the kinase activity of Akt. PMID- 19815944 TI - 2-Methyl-6-phenylethynyl-pyridine (MPEP), a non-competitive mGluR5 antagonist, differentially affects the anticonvulsant activity of four conventional antiepileptic drugs against amygdala-kindled seizures in rats. AB - 2-Methyl-6-phenylethynyl-pyridine (MPEP), a selective noncompetitive mGluR5 antagonist, influences the action of conventional antiepileptic drugs in amygdala kindled seizures in rats. MPEP alone (up to 40 mg/kg) did not affect any seizure parameter. Moreover, the common treatment of MPEP with either carbamazepine or phenytoin (administered at subeffective doses) did not result in any anticonvulsant action in kindled rats. However, when combined with subprotective doses of valproate or phenobarbital, MPEP significantly shortened seizure and afterdischarge durations. Importantly, combinations of MPEP with the two antiepileptics did not have the adverse effects of impaired motor performance or long-term memory in rats. Our data indicate that MPEP may positively interact with some conventional antiepileptic drugs in the amygdala-kindling model of complex partial seizures. PMID- 19815945 TI - Bradykinin receptor antagonists and cyclooxygenase inhibitors in vincristine- and streptozotocin-induced hyperalgesia. AB - Pain that accompanies neuropathy is difficult to treat. Analgesics administered as monotherapies possess low activities in relieving this kind of pain. The effect of the simultaneous administration of indomethacin (a preferential inhibitor of cyclooxygenase-1; COX-1) or celecoxib (a relatively selective inhibitor of cyclooxygenase-2; COX-2), with selective antagonists of bradykinin(2) (B(2)) bradykinin(1) (B(1)) receptors (HOE 140 or des-Arg(10)-HOE 140) on the alleviation of diabetic and toxic neuropathic pain was investigated. Pretreatment with indomethacin (0.1 mg/kg, sc) increased the antihyperalgesic activity of low daily doses of HOE 140 or des-Arg(10)HOE 140 (70 nmol/kg, ip) in a diabetic (streptozotocin(STZ)-induced) neuropathy/hyperalgesia experimental model. Premedication with celecoxib before HOE 140 or des-Arg(10)HOE 140 administration resulted in a gradual reduction of STZ hyperalgesia. Furthermore, on days 23-24, almost complete abolishment of STZ hyperalgesia was observed. After cessation of drug administration, hyperalgesia quickly returned to the baseline threshold. The results of this study suggest that inhibitors of cyclooxygenases can increase the antihyperalgesic activity of selective antagonists of B(2) and B(1) receptors in diabetic and toxic neuropathic pain models. These observations may be clinically relevant. PMID- 19815946 TI - Optimization of lidocaine application in tumescent local anesthesia. AB - Tumescent local anesthesia is based upon the infusion of large volumes of neutralized anesthetic solutions, mainly lidocaine, at very low concentrations. This results in the paralysis of sensory nerve endings and minute nerve twigs, leading to a reduction in pain. The aim of this study was to assess the safety of lidocaine application in tumescent local anesthesia on different regions of patient's bodies. Measures of safety included the analysis of lidocaine concentrations and its pharmacokinetic parameters. In total, 48 patients were infused with tumescent anesthesia in the hypogastrium, buttocks and thighs, axillae, breasts, trunk, and face and neck areas. Lidocaine was infused in doses ranging from 5.2-40 mg/kg b.w., and in concentrations of 0.05% (hypogastrium, buttocks, thighs) or 0.1-0.15% (axillae, breasts, trunk, face, neck), using a total amount of 300-3200 mg. As the peak lidocaine concentration did not exceed 5 microg/ml (commonly known as the toxic threshold), the results of our study indicate that the doses used (not exceeding 40 mg/kg b.w.) are completely safe for patients undergoing tumescent anesthesia in different body areas. The observation of statistically significant correlations between both the dose and the total amount of lidocaine administered and its peak plasma concentration, together with the lack of correlations between the dose and the amount and the time taken to reach peak concentration, allows the safety of each anesthetic dose to be predicted. An analysis of the heterogeneous dynamics of lidocaine plasma concentration changes in tumescent anesthesia in different body areas indicates that both the rates and the degrees of absorption and elimination depend on the area of infiltration; this is in turn related to the vascularization of any given area. The study of lidocaine concentration and pharmacokinetic parameters also showed that there may potentially be a higher risk of a large anesthetic concentration developing within a short period of time during anesthesia of the upper parts of the body. During tumescent anesthesia, significantly higher plasma concentrations of lidocaine were observed in the face and neck than in the hypogastrium, buttocks and thighs, axillae, breast and trunk 0.5 to 4 h after its infusion. This indicates the need for carefully conducted patient observations immediately after infiltration into the aforementioned areas. PMID- 19815947 TI - Effect of HMG-CoA (3-hydroxy-3-methylglutaryl-CoA) reductase inhibitors on the concentration of insulin-like growth factor-1 (IGF-1) in hypercholesterolemic patients. AB - Studies have shown that HMG-CoA reductase inhibitors (statins) play an important role in the prevention and treatment of atherosclerosis and hyperlipidemia. The aim of this study was to investigate the effect of 3-month treatment with simvastatin on serum levels of Insulin-Like Growth Factor-1 (IGF-1) in patients with diagnosed hypercholesterolemia. In total, 156 patients with hypercholesterolemia were recruited for the study. The inclusion criteria for this study were designed to allow the enrollment of a representative group of patients for cytokine studies. The patients were divided into two groups: (1) patients with a mild-to-moderate risk of heart disease, who had total cholesterol (TC) < 300 mg/dl (7.8 mmol/l), LDL-cholesterol < 210 mg/dl (5.4 mmol/l), and who lacked risk factors for coronary artery disease (CAD) after treatment with a diet for 3 months; (2) patients with a high-to-very high risk of CAD, who had TC > 300 mg/dl (7.8 mmol/l), LDL-cholesterol > 210 mg/dl (5.4 mmol/l), and at least two risk factors for CAD after treatment with a diet and administration of simvastatin (20 mg/day) for a three month period. The control group consisted of ten healthy volunteers who each had a normal lipid profile. Total cholesterol, LDL-cholesterol and IGF-1 concentrations were measured at baseline and either after six months of dietary supplementation (first group) or after three months of dietary supplementation and three months of simvastatin treatment (second group). CONCLUSIONS: In patients with mild-to-moderate risk of CAD, a decreased serum concentration of IGF-1 was observed three months after beginning a low-fat diet. However, no changes in the serum concentration of IGF-1 were noted in patients with high-to-very high risk of CAD. Additional three-month treatment with simvastatin decreased the serum concentration of IGF-1. PMID- 19815948 TI - Beneficial effects of L-arginine against diabetes-induced oxidative stress in gastrointestinal tissues in rats. AB - Oxidative stress occurs in diabetic patients and experimental models of diabetes. The ability of L-arginine to ameliorate oxidative stress after treatment with alloxan was investigated in rats. Adult male rats were injected intraperitoneally with multiple doses of alloxan to produce experimental oxidative stress characteristic of diabetes mellitus. The rats were maintained in this state for eight weeks. Rats were decapitated and gastrointestinal tissues were isolated. The results were interpreted and the significance was analyzed using unpaired, two-tailed Student's t tests and two-way repeated measures ANOVA. Hyperglycemia was observed in the plasma after three days of alloxan treatment. This was associated with a depression of glutathione (GSH) concentration as well as superoxide dismutase (SOD) and catalase (CAT) activities in the pylorus and ileum. In addition, the malonyldialdehyde (MDA) levels were significantly elevated, indicating increased lipid peroxidation and oxidative stress in the same tissues. L-arginine supplementation (0.15 mg/ml) through drinking water until eight weeks after alloxan injection significantly ameliorated the oxidative stress, as evidenced by lower MDA levels as well as higher levels of endogenous GSH, SOD, and CAT (p < 0.001). These effects were paralleled by marked protection and prophylaxis against alloxan-induced hyperglycemia. Thus, exogenously administered L-arginine might improve the clinical manifestation of diabetes mellitus and decrease the oxidative stress in the gastrointestinal tract. In addition, the study supports the beneficial effects of L-arginine. These effects might be attributed to its direct, NO-dependent antioxidant capacity and/or NO independent pathways. PMID- 19815949 TI - Accelerated thrombus lysis in the blood of plasminogen activator inhibitor deficient mice is inhibited by PAI-1 with a very long half-life. AB - Patients with defective plasminogen activator inhibitor protein (PAI-1) or with PAI-1 deficiency can experience hemorrhage as a result of a hyperfibrinolysis. In these patients, a normal thrombus forms, but endogenous lysis is unchecked as tissue plasminogen activator is unopposed. Treatment includes anti-fibrinolytic agents, including oral tranexamic acid. Another treatment option is the administration of PAI-1, but this serpin rapidly inactivates itself. We have developed a mutant plasminogen activator inhibitor with a very long half life (VLHL PAI-1, t1/2>700 h). Here we investigate VLHL PAI-1 effects in the blood of PAI-1 deficient mice, as a model of human disease. Using a thrombelastograph, we found that blood clots of PAI-1 knockout mice were lysed much more quickly than wild type mice. Additionally, blood clots had less shear elastic modulus strength than clots of normal animals. VLHL PAI-1 treatment of PAI-1 deficient mice extended or prevented thrombus lysis and increased clot strength in a concentration dependent fashion. These two parameters determine the extent of thrombus growth and regression; thus, further testing is anticipated to confirm the effectiveness of plasminogen activator inhibitor with a very long half life in an in vivo model and we hope that this protein can be effective in human PAI-1 deficiency disorder. PMID- 19815950 TI - Effects of taurine on contractions of the porcine coronary artery. AB - The actions and mechanisms of taurine on vascular contractions have been studied in the isolated porcine coronary artery. Taurine depressed histamine-, serotonin , KCl- and CaCl(2)-induced contractions in a concentration-dependent manner, with maximal contractions being depressed by 43.4%, 46.2%, 33.3% and 43.3%, respectively. Taurine relaxed arterial rings that were precontracted by either 30mM KCl or 0.3 muM U46619, a thromboxane A(2) analog, in a concentration dependent manner, and the maximal relaxations were 39.4% and 38.7%, respectively. The vasorelaxations were nearly abolished by pretreatment with either the inward rectifier K(+) channel (K(IR)) inhibitor, BaCl(2), or the ATP sensitive K(+) channel (K(ATP)) inhibitor, glibenclamide, and were attenuated by the Ca(2+) activated K(+) channel (K(Ca)) inhibitor tetraethylammonium. Denudation of the endothelium, and treatment with the nitric oxide synthase inhibitor, L-NAME, the cyclooxygenase inhibitor, indomethacin, or the voltage gated K(+) channel (K(V)) inhibitor 4-aminopyridine did not affect the relaxation. The present results show that taurine antagonizes and relaxes the contractions of the porcine coronary artery, and suggest that the activation of K(IR), K(ATP) and K(Ca) may be involved in taurine-induced relaxation of the porcine coronary artery. PMID- 19815951 TI - Association of the MDR1 (ABCB1) gene 3435C>T polymorphism with male infertility. AB - Infertility is a common problem affecting one in six couples, and in 30% of infertile couples, the male factor is a major cause due to defective sperm quality. P-glycoprotein (P-gp), a product of the MDR1 (ABCB1) gene, may be a link between genetic and environmental factors contributing to the development of male infertility because pesticides (P-gp substrates) are well established factors of male infertility. The aim of the present study was to examine the effect of the MDR1 gene 3435C>T polymorphism on male infertility. In total, 162 male patients undergoing semen analysis due to initial infertility workup were included in the study. The control group consisted of 191 healthy males with proven fertility. MDR1 3435C>T genotyping was performed by the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. Assessment of MDR1 genotypes among the infertile men showed that 17.9% of subjects were carriers of the CC genotype, 58.0% were CT and 24.1% were TT. Among fertile men, 30.4% of subjects were characterised by the CC genotype, 49.7% were CT and 19.9% were TT. In addition, the frequency of carriers of at least one T allele (i.e., CT and TT genotypes) among infertile and fertile subjects was 82.1% and 69.6%, respectively. The risk of infertility was significantly elevated by two-fold in individuals carrying at least one T allele (CT and TT genotypes: p = 0.009, OR = 2.00, 95% CI: 1.20-3.32). Furthermore, this elevated risk was still found when considering each of the CT and TT genotypes alone (TT genotype: p = 0.027, OR = 2.05, 95% CI: 1.09-3.86; CT genotype: p = 0.013, OR = 1.98, 95% CI: 1.16-3.36). This preliminary report suggests that P-gp may play some role in male infertility, mediating detrimental effects of environmental factors. PMID- 19815952 TI - Effects of escin on acute inflammation and the immune system in mice. AB - Escin has been used extensively to treat chronic venous insufficiency, hemorrhoids, and edema resulting from cerebral ischemic damage, trauma or operation. However, no studies have looked at the anti-inflammatory properties of escin administered by intravenous injection, and it is still not clear whether escin has an effect on the immune system. This study seeks to investigate the time-dependent anti-inflammatory properties of escin and its effect on the immune system. The anti-inflammatory effect of escin was observed in carrageenan-induced paw edema and acetic acid-induced capillary permeability in mice. The immunopharmacological effects of escin were evaluated by spleen index (SI), thymus index (TI), proliferative capacity of splenocytes (PS), lymphocyte count (LC), serum TNF-alpha levels, and phagocytic rate (PR) in mice. Escin treatment showed a significant anti-inflammatory effect, similar to that seen with dexamethasone treatment. However, the duration of the anti-inflammatory response was longer with escin treatment than with dexamethasone treatment. The results also demonstrated that escin had no significant effects on SI, TI, LC, PS, TNF alpha levels, and PR. The findings suggest that escin is a potent anti inflammatory drug with long-lasting anti-inflammatory effects and without any immunosuppressive effects. PMID- 19815953 TI - Endogenous steroids are responsible for lactoferrin-induced myelopoiesis in mice. AB - Our previous study revealed that lactoferrin (LF) significantly increases mobilization of the myelocytic lineage in mice. The aim of our current investigation was to determine whether activation of the hypothalamic-pituitary adrenal axis contributes to this phenomenon. We found that intravenous (iv) injection of LF (10 mg) caused a 48.8% increase in the circulating blood leukocyte count and increased the proportion of the myelocytic lineage (band forms, 10-fold and neutrophils, 2-fold) 24 h post injection. The content of the myelocytic lineage (myelocytes, metamyelocytes, bands and neutrophils) in bone marrow rose from 51.6 to 63.4%. In addition, administration of LF led to a decrease in total thymocyte number by 41.6%. Analogous changes in cell types and numbers in adrenalectomized mice following LF injection were minor. Mifepristone, a blocker of steroid receptors, reversed the effects of LF on leukocyte cell number and bone marrow cell composition. Finally, we showed that LF induced a rise in the serum levels of corticosterone in control but not adrenalectomized mice.We conclude that LF-induced upregulation of endogenous steroid levels is responsible for the stimulation of myelopoiesis. PMID- 19815954 TI - Fluoroquinolones lower constitutive H2AX and ATM phosphorylation in TK6 lymphoblastoid cells via modulation of the intracellular redox status. AB - Accumulation of reactive oxygen species (ROS)-induced damage and mutations in the genomic DNA is considered the primary etiology of aging and age-related pathologies including cancer. Strategies aimed at slowing these conditions often involve protecting against oxidative DNA damage via modulation of the intracellular redox state. Recently, a biomarker of DNA double-strand breaks (DSBs), serine 139-phosphorylated histone H2AX (gammaH2AX), and its upstream mediator, activated PI-3-related kinase, ATM (ATM(P1981)), were shown to be constitutively expressed in cells and modulated by antioxidant treatment. Thus, both constitutive histone H2AX phosphorylation (CHP) and constitutive ATM activation (CAA) are thought to reflect a cell's response to endogenous ROS induced DSBs. In the present study, we investigated the effects of a battery of fluoroquinolone (FQ) compounds, namely ciprofloxacin, enrofloxacin, gatifloxacin, lomefloxacin and ofloxacin, on CHP and CAA in human TK6 lymphoblastoid cells. All FQs tested reduced CHP and CAA compared to controls following 6 and 24 h treatment with CAA being more sensitive to their effects at both time points. In addition, intracellular ROS levels and mitochondrial activities were also lowered in FQ-treated cells at 6 and 24 h.We presume that FQs mediate this effect via a combination of ROS-scavenging and mitochondrial suppression and therefore may protect against the onset or may slow the progression of numerous oxidative pathophysiological conditions. PMID- 19815956 TI - Synthesis and pharmacological properties of a new fluorescent opioid peptide analog. AB - Biphalin, is a palindromic peptide [(Tyr-D-Ala-Gly-Phe-NH-)2] in which two opioid pharmacophores are connected "tail-to-tail". This peptide displays a broad affinity for all opioid receptors (mu, delta and kappa) as well as exceptionally high antinociceptive activity. Previous structure-activity studies demonstrated that one of the biphalin pharmacophores could be substituted with a hydrophobic group without significant loss of receptor affinity. This paper reports the pharmacological properties of a new analog in which one pharmacophore of biphalin was replaced with fluorescent 7-succinylamido-4-methyl-coumarin. The resulting compound displays an affinity for mu opioid receptors that is a delta opioid receptor comparable to biphalin but with an affinity that is over a hundred times lower. This mu opioid selective fluorescent peptide analog could be applied in pharmacokinetic and pharmacodynamic studies of biphalin related analogs. PMID- 19815955 TI - Circadian time-dependent chemopreventive potential of withaferin-A in 7,12 dimethylbenz[a]anthracene-induced oral carcinogenesis. AB - Circadian time-dependent treatment with chemotherapeutic drugs (chronotherapy) optimizes the therapeutic index by maximizing treatment efficacy and minimizing toxicity. The circadian time-dependent chemopreventive and anti-lipid peroxidative efficacy of withaferin-A in 7,12-dimethylbenz[a]anthracene (DMBA) induced hamster buccal pouch carcinogenesis was investigated in the present study. We induced oral squamous cell carcinoma in the buccal pouches of golden Syrian hamsters during the day (4:00, 8:00, 12:00, 16:00, 20:00 and 24:00) by application of DMBA three times per week for 14 weeks. The circadian time dependent tumor incidence, volume and burden were observed in hamsters treated with either DMBA alone or DMBA + withaferin-A. The circadian pattern of lipid peroxidation by-products, as measured by the formation of thiobarbituric acid reactive substances (TBARS) and enzymatic antioxidants [superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], was also analyzed in the buccal mucosa of DMBA-treated hamsters. We found the highest incidence of tumor formation at 24.00 h in hamsters treated with DMBA alone as compared to other experimental groups. Circadian dysregulation of lipid peroxidation and antioxidant status was observed in DMBA-treated animals as compared to control animals. Oral (po) administration of withaferin-A (20 mg/kg) completely prevented the formation of tumors between 8.00 h and 12.00 h and synchronized the status of lipid peroxidation and antioxidants in the buccal mucosa of hamsters treated with DMBA alone. Also, oral administration of withaferin-A to DMBA-treated animals significantly reduced the formation of tumors and synchronized the status of lipid peroxidation and antioxidants in the rest of the time intervals. Our study thus suggests that withaferin-A has significant chemopreventive and anti-lipid peroxidative potential in DMBA-induced oral carcinogenesis, probably by interfering with DMBA-induced abnormal cell proliferation in the buccal mucosa. PMID- 19815957 TI - Influence of orphenadrine upon the protective activity of various antiepileptics in the maximal electroshock-induced convulsions in mice. AB - Orphenadrine is an anticholinergic drug used in the treatment of Parkinson's disease, and is also known to exert nonspecific antagonistic activity at the phencyclidine binding site of the N-methyl-D-aspartate (NMDA) receptor. The aim of this study was to assess the anticonvulsant properties of orphenadrine and to evaluate its effect on the anticonvulsant activity of antiepileptic drugs against maximal electroshock-induced seizures in mice. Orphenadrine given at a dose of 5.65 mg/kg elevated the electrical seizure threshold from 5.7 (5.4-6.1) to 6.8 (6.3-7.3) mA, while a dose of 2.8 mg/kg was ineffective. The ED(50) values of orphenadrine administered 10, 30 and 120 min before maximal electroshock-induced convulsions were 16.8 (11.3-25.1), 17.8 (15.7-20.0) and 25.6 (23.3-28.3) mg/kg, respectively. Orphenadrine at a sub-threshold dose of 2.8 mg/kg significantly enhanced the anticonvulsant activity of valproate by reducing its ED(50) value from 315.8 (270.0-369.4) to 245.9 (207.1-292.0) mg/kg without affecting the free plasma levels of valproate. However, orphenadrine failed to enhance the protective activity of carbamazepine, phenytoin, phenobarbital, lamotrigine, topiramate, or oxcarbazepine against maximal electroshock-induced seizures. PMID- 19815958 TI - Characterization of acute adverse-effect profiles of selected antiepileptic drugs in the grip-strength test in mice. AB - The aim of this study was to assess the acute adverse effects (neurotoxic) of several antiepileptic drugs (clonazepam, lamotrigine, oxcarbazepine, phenytoin, phenobarbital and topiramate) by measuring skeletal muscular strength in mice using the grip-strength test. Linear regression analysis of grip-strength in relation to drug dose-response allowed us to determine D(50) values, the dosages of antiepileptic drugs that reduced grip-strength in mice by 50% compared to control animals. Each of the antiepileptic drugs studied reduced skeletal muscular strength in mice in a dose-dependent manner. The D(50) for clonazepam was 31.7 mg/kg, lamotrigine -47.7 mg/kg, oxcarbazepine -87.3 mg/kg, phenobarbital -128.7 mg/kg, phenytoin -69.7 mg/kg, and topiramate -509.5 mg/kg. In conclusion, the grip-strength test can aid in evaluating acute adverse effects of drugs with respect to their influence on muscular strength in experimental animals. PMID- 19815959 TI - Neuroprotective effect of N-acetylcysteine in neurons exposed to arachidonic acid during simulated ischemia in vitro. AB - The aim of the study was to assess neuroprotective effects of N-acetylcysteine (NAC; 100-200 microM) on cultured cortical neurons exposed to arachidonic acid (AA, 10 microM) during ischemia (oxygen-glucose deprivation). Ischemic conditions decreased neuron viability to 41-47% of normoxic controls; co-exposure with arachidonic acid further attenuated neuron viability to 36.73% after 24 h. Separate exposure to arachidonic acid in normoxia or to ischemic conditions only, increased the number of apoptotic nuclei to 33.56% or 36.78%, respectively. Combined exposure to arachidonic acid and ischemia increased apoptosis frequency to 62.20%. NAC (200 microM) decreased the number of apoptotic nuclei in normoxia in control and arachidonic acid exposed cells. NAC also decreased apoptosis frequency in ischemia to 14%. In neurons exposed to arachidonic acid and ischemic conditions, 100 and 200 microM NAC reduced apoptosis to 24.99% and 19.48%, respectively. NAC provided protection to neurons from toxicity due to arachidonic acid, ischemia and exposure to arachidonic acid in ischemic conditions. PMID- 19815960 TI - Kynurenic acid protects against the homocysteine-induced impairment of endothelial cells. AB - Kynurenic acid (KYNA) is a tryptophan metabolite produced in the kynurenine pathway. In the central nervous system, KYNA exerts neuroprotective and anticonvulsant effects by mechanisms associated with its antagonist activity against the ionotropic glutamate and alpha-7 nicotinic receptors. Its presence has been documented not only in cerebrospinal fluid and brain tissue, but also in the periphery. However, KYNA's function outside the brain has not been fully elucidated. In this study, experiments performed on bovine aorta endothelial cell cultures showed for the first time that KYNA exerts a protective activity against the homocysteine-induced impairment of endothelial cells. The addition of KYNA significantly increased endothelial cell migration and proliferation, which is diminished by homocysteine. KYNA also protected cells against homocysteine induced cytotoxicity. Our data suggest that increasing KYNA levels in blood vessels may have a significant impact on the endothelium in hyperhomocysteinemia. PMID- 19815961 TI - Subcutaneous metastasis of prostate cancer. AB - OBJECTIVES: Prostate cancer tends to spread to regional lymph nodes and bone, and, to a lesser degree, to lung, liver, and brain. Metastases in other locations are exceptional. To review the frequency and clinical characteristics of metastasis to subcutaneous cellular tissue in adenocarcinoma of the prostate. METHODS: The case of a 71-year-old man diagnosed of adenocarcinoma of the prostate is reported. The patient underwent radical prostatectomy and artificial sphincter for stress urinary incontinence. RESULTS: During follow-up the patient showed biochemical progression, local recurrence, and bone metastasis. The disease metastasized in the subcutaneous tissue around the reservoir of the artificial sphincter. CONCLUSIONS: Prostate cancer is highly prevalent in our part of the world. The clinical finding of metastasis in organs other than bone or regional lymph nodes is accompanied by a short survival. Metastases in subcutaneous tissue may be underdiagnosed due to its indolent clinical course and possible absence of PSA elevation. In any case, subcutaneous metastases have an unfavorable prognosis. PMID- 19815962 TI - Median raphe cyst. Report of two cases and literature review. AB - OBJECTIVES: To report 2 cases of median raphe cysts, 1 in the penis and the other in the perineum. METHOD: Two cases of median raphe cyst are described; the first was treated by surgery and the second required no treatment. RESULTS: The surgical patient experienced no complications and was asymptomatic with no recurrence at 2 years. CONCLUSIONS: Median raphe cysts are a rare, benign condition of uncertain etiology. Treatment consists on simple excision. PMID- 19815963 TI - Persistence of an infected urachus presenting as acute abdominal pain. Case report. AB - OBJECTIVES: We report a case of urachal remnant disease and review the literature. METHODS/RESULTS: We present the case of an urachal cyst in a 13-year old patient who was admitted to the emergency department with acute abdominal pain. Differential diagnosis of his symptoms was made with other diseases such as appendicitis and inflammatory bowel disease. CONCLUSIONS: Urachal remnant diseases are rare and they usually present during the neonatal period with fever and wet navel, lower abdominal pain around the middle line, palpable mass and urination symptoms with or without urinary infections. The presentation as acute abdominal pain in an older child is less common, and its differential diagnosis must be performed with other abdominal or pelvic acute diseases. The most appropriate imaging technique is an ultrasound exam. PMID- 19815964 TI - Giant cystic degeneration of the rete testis. AB - OBJECTIVES: We present a case of giant cyst of the rete-testis. METHODS/RESULTS: 85 year-old patient on follow -up for prostate cancer with maximum androgen blockade (MAB) treatment consults for a left hemiscrotum increase in size over a 2-month period. We performed bilateral orchiectomy confirming the histopathological diagnosis of cystic dilatation of the rete-testis sized 11 x 11 x 9 cm. CONCLUSIONS: This case of cystic degeneration of the rete-testis, with a size out of common (11 x 11 x 9 cm versus medium size in the literature: 3 x 3 x 3 cm), could be related with an androgen-estrogen misbalance caused by a MAB in a prostate cancer context. PMID- 19815965 TI - Urological conservative management of a patient with tuberous sclerosis complex (Bourneville disease). AB - OBJECTIVE: To report the clinical and radiological data of a 23-year-woman with bilateral angiomyolipoma and Tuberous Sclerosis with conservative treatment. METHODS: The patient underwent conservative urological management, based on clinical and complementary tests data (ultrasonography, CT scanning, excretory urogram, and laboratory). Monitoring was carried out in external consultation by the urooncology group. In some occasions she was hospitalized due to distant geographical factors. RESULTS: This patient provides long-term outcome of AML with Tuberous Sclerosis followed by repeated evaluation to define the conservative management or not. Ultrasound and CT scan were two basic studies to maintain conservative treatment, as well as a good clinical evaluation. CONCLUSIONS: Renal lesions associated with tuberous sclerosis are frequent and serious. Monitoring should be based on a careful monitoring since the renal lesions are the second death cause after the lesions of the nervous system. This case report provides long-term outcome of AML with tuberous sclerosis, that can be followed expectantly with repeated evaluations to define the clinical significance. PMID- 19815966 TI - Health and quality of life in urology: issues in general urology and urological oncology. AB - OBJECTIVES: Frequently, the term "quality of life" has been used to justify personal and professional decisions in all fields of medicine. Nowadays, quality of life studies are based on development and validation of sensitive measures of patient outcomes, incorporating functional status and perceived health status. Thus, quality of life has become an outcome as important as survival and effectiveness. METHODS: A systematic review using Pubmed and Medline was performed, searching for papers concerning health related quality of life and urology. The most relevant articles where questionnaires and interviews were described and validated were listed. RESULTS: Based on psychometric properties, a search between 1970 and 2007 identified a total of 25 recommendable articles with generic inventories and specific modules that have been developed, validated and used in clinical practice or research. Historical aspects, quality of life concepts, validation of questionnaires and structured interviews, and most used instruments in generic health-related quality of life, general urology and urological oncology have been discussed. CONCLUSIONS: A brief review of historic background of health related quality of life and urology was performed. PMID- 19815967 TI - Fournier's gangrene: our experience in 5 years, bibliographic review and assessment of the Fournier's gangrene severity index. AB - OBJECTIVES: Fournier's gangrene is a devastating infection, which includes the genital, perineal and/or perianal regions. It is potentially fatal, and affects any age and gender. The severity index for Fournier's gangrene has been described; it is useful for evaluating the prognosis of these patients. Our goal is to report our experience with this disease over the past 5 years and evaluate the index in retrospect. METHODS: We analyzed medical records of patients with Fournier gangrene over the last 5 years at the University Hospital "Dr. Jose E. Gonzalez". RESULTS: We reviewed 50 cases, male gender was predominant (96%), mean age 47.5 years, diabetes mellitus was found in 80%, neurogenic bladder in 10%, 2% HIV positive. The most frequent sites of origin of infection were scrotum (52%) and perineum (38%), the most common pathogen E. coli and Enterococcus faecalis (48 and 28% respectively). The death rate was 12%. The average severity index was 5.64. DISCUSSION: In our hospital, Fournier's gangrene is rare. Nevertheless, there is a rapid diagnosis protocol and therapeutic management is performed immediately. Until now, the immediate surgical treatment and early initiation of antibiotic therapy remains the best therapeutic option. CONCLUSION: There is a relationship between the index of severity and patient survival, which may become a useful parameter in evaluating these patients. PMID- 19815968 TI - Outcomes of maximal androgen blockade in prostate cancer patients at a health area with type 2 reference hospital. Part 2. Quality of life: application of EORTC QLQ-PR25 instrument and global results. Quality-of-life adjusted survival. Pharmaceutical expenses and cost-utility. AB - OBJECTIVES: To study quality of life in patients from the health area of Zamora diagnosed of prostate adenocarcinoma between 2000-2005 treated with maximal androgen blockade (MAB). To evaluate the pharmacoeconomics of the treatment. METHODS: Basal, 12-month, 24-month and 36-month application of the health-related quality of life measurement instrument EORTC QLQ-C30 to the population sample (n= 111), as well as a control sample (n= 100). Comparative study of outcomes: between groups; between different time measurements in MAB patients; and inter categories/ -intervals of some variables in patients with hormonal deprivation therapy (third year of follow-up). Analysis of health-related quality of life global outcomes (QLQ-C30 + QLQ-PR25). Description of the pharmaceutical expenses in androgen blockade patients. Cost-utility analysis by means of quality-adjusted life years (QALYs) obtained using preference-based weighted index from the EUROQOL 5-D tool. RESULTS: Hot flushes and sexual field worsening as mean expressions related to hormonal suppression. Treated patients had worse subjective perception of health condition and quality of life, in opposition to non-tumoral individuals. Positive bone scan, was a negative-influence factor on quality of life. Most patients undergoing MAB needed cost-utility figures of less than 5000 Euros/ QALY. CONCLUSIONS: There was a negative initial repercussion of MAB on quality of life, although tinged according to the different fields studied. The economic impact of this therapy on overall pharmaceutical expenses is relative. PMID- 19815969 TI - SolidFlex--the fourth generation in endoscopy. AB - OBJECTIVES: To design and develop the fourth generation in endoscopy after the rigid, semirigid and flexible instruments. This next generation technology has been named SolidFlex, and it was designed by a surgeon based on many years of research with the objective to provide easier and faster solutions for physicians when performing endoscopic procedures. METHODS: This new endoscope called SolidFlex has been brought to light with the latest technology to provide surgeons of every age the capabilities to carry out any possible endoscopic procedure. SolidFlex is state-of-the-art in both design and construction, giving the the weight of the usual flexible endoscope. The Instrument is ergonomic, intuitive (instinctive perception) and coherent, meaning that the handle control moves the instrument's active flexible section towards predictable, pre-selected areas, driving the procedure fluidly throughout. It provides the stability of a rigid and semirigid instrument with the exploration capabilities of the flexible endoscope in order to achieve the best means of rendering a patient 100% stone free. RESULTS: The SolidFlex ureteroscope prototype has retrieved urinary stones in 9 patients and 11 renal units. These procedures were divided in two groups: Group I, pre-stented after double "J" withdrawal for a previous renal colic episode. Group II, during the acute renal colic episode. Ureteral dilatation was carried out only in Group II and after dilatation in both groups, insertion up to the renal pelvis or the area were the obstruction was present, took only a matter of 15 seconds at the most. All procedures could be completed in each group and stones in the renal papilla from 200 nm, to 4 mm in any given papilla were removed either by laser energy, retrieved with a basket or both. Patients did not experience any pain or major discomfort after the procedure, and no analgesic medication was required, nor requested by any of the patients. CONCLUSIONS: The new SolidFlex technology has been tested in surgery and proven to solve most of the difficulties faced by surgeons when performing endoscopic surgery. The surgeon can perform surgery comfortably seated, so that he/she can easily introduce and maneuver the instrument, and introduce operating tools without help or coordination from anyone else during critical moments such as stone extraction. A urologist made the SolidFlex research and development for the benefit of physicians and patients, and it does solve many of the issues and limitations that surgeons face today with the rigid, semi-rigid and flexible endoscope technology. PMID- 19815970 TI - Electrospinning-aligned and random polydioxanone-polycaprolactone-silk fibroin blended scaffolds: geometry for a vascular matrix. AB - Extracellular matrices are arranged with a specific geometry based on tissue type and mechanical stimulus. For blood vessels in the body, preferential alignment of fibers is in the direction of repetitive force. Electrospinning is a controllable process which can result in fiber alignment and randomization depending on the parameters utilized. In this study, arterial grafts composed of polycaprolactone (PCL), polydioxanone (PDO) and silk fibroin in blends of 100:0 and 50:50 for both PCL:silk and PDO:silk were investigated to determine if fibers could be controllably aligned using a mandrel rotational speed ranging from 500 to 8000 revolutions per minute (RPM). Results revealed that large- and small-diameter mandrels produced different degrees of fiber alignment based on a fast Fourier transform of scanning electron microscope images. Uniaxial tensile testing further demonstrated scaffold anisotropy through changes in peak stress, modulus and strain at break at mandrel rotational speeds of 500 and 8000 RPM, causing peak stress and modulus for PCL to increase 5- and 4.5-fold, respectively, as rotational speed increased. Additional mechanical testing was performed on grafts using dynamic compliance, burst strength and longitudinal strength displaying that grafts electrospun at higher rotational rates produced stiffer conduits which had lower compliance and higher burst strength compared to the lower mandrel rotational rate. Scaffold properties were found to depend on several parameters in the electrospinning process: mandrel rotational rate, polymer type, and mandrel size. Vascular scaffold design under anisotropic conditions provided interesting insights and warrants further investigation. PMID- 19815984 TI - Effect of donor cell types on developmental potential of cattle (Bos taurus) and swamp buffalo (Bubalus bubalis) cloned embryos. AB - This study investigated the effect of donor cell types on the developmental potential and quality of cloned swamp buffalo embryos in comparison with cloned cattle embryos. Fetal fibroblasts (FFs), ear fibroblasts (EFs), granulosa cells (GCs) and cumulus cells (CCs) were used as the donor cells in both buffalo and cattle. The cloned cattle or buffalo embryos were produced by fusion of the individual donor cells with enucleated cattle or buffalo oocytes, respectively. The reconstructed (cloned) embryos and in vitro matured oocytes without enucleation were parthenogenetically activated (PA) and cultured for 7 days. Their developmental ability to the blastocyst stage was evaluated. The total number of trophectoderm (TE) and inner cell mass (ICM) cells and the ICM ratio in each blastocyst was determined by differential staining as an indicator of embryo quality. The fusion rate of CCs with enucleated oocytes was significantly lower than for those of other donor cell types both in cattle and buffalo. The rates of cleavage and development to the 8-cell, morula and blastocyst stages of cloned embryos derived from all donor cell types did not significantly differ within the same species. However, the cleavage rate of cloned cattle embryos derived from FFs was significantly higher than those of cattle PA and cloned buffalo embryos. The blastocyst rates of cloned cattle embryos, except for the ones derived from CCs, were significantly higher than those of cloned buffalo embryos. In buffalo, only cloned embryos derived from CCs showed a significantly higher blastocyst rate than that of PA embryos. In contrast, all the cloned cattle embryos showed significantly higher blastocyst rates than that of PA embryos. There was no difference in ICM ratio among any of the blastocysts derived from any of the donor cell types and PA embryos in both species. FFs, EFs, GCs and CCs had similar potentials to support development of cloned cattle and buffalo embryos to the blastocyst stage with the same quality. PMID- 19815985 TI - A comparison of cryotop and solid surface vitrification methods for the cryopreservation of in vitro matured bovine oocytes. AB - The aim of the present study was to compare the efficacies of the cooling systems of the solid surface (SSV) and Cryotop vitrification methods for cryopreservation of bovine oocytes at the metaphase II stage. The effects of vitrification on oocyte viability, in vitro fertilization (IVF), pronucleus formation and subsequent in vitro development were assessed. In vitro matured (IVM) bovine oocytes were subjected to equilibration and vitrification solutions according to the SSV method, and then the oocytes were vitrified either by dropping onto a cold dry metal surface (SSV group) or by plunging into liquid nitrogen on Cryotop sheets (Cryotop group). Warming was conducted according to the SSV method. Some oocytes were subjected to the cryoprotectants and warming regimen without cooling (Solution control group). The live/dead status of oocytes was evaluated by fluorescein diacetate staining. Live oocytes were subjected to IVF, and the resultant embryos were cultured in vitro. The rates of live oocytes were similar among the Fresh control, Solution control, SSV and Cryotop groups. There was no difference in the rates of fertilization, pronuclear formation and monospermy among these groups. The cleavage rates in the SSV and Cryotop groups (41.6 and 53.2%, respectively) were significantly lower than those in the Fresh control and Solution control groups (65.9 and 61.3%, respectively). The blastocyst rates in SSV and Cryotop groups did not differ (10.3 and 12.8%, respectively); however, they were significantly lower than those in the Fresh control and Solution control groups (36.4 and 24.8%, respectively). The inner cell mass, trophectoderm and total cell numbers in blastocysts did not differ significantly among the Fresh control, Solution control, SSV and Cryotop groups. Our results indicate that IVM bovine oocytes could be cryopreserved successfully using the cooling systems of the Cryotop and Solid Surface Vitrification methods with similar efficacy. PMID- 19815986 TI - Gene silencing of DNA methyltransferases by RNA interference in bovine fibroblast cells. AB - A highly methylated genome, like a somatic donor cell, is observed in somatic cell nuclear transfer (SCNT) embryos. The aberrant DNA methylation status causes global gene expression failure, resulting in low developmental competence of SCNT embryos. In addition, recent studies have uncovered the relationship between DNA methylation status and reprogramming efficiency. Because DNA methylation is performed by DNA methyltransferases (DNMTs), developing a technique which specifically inhibits DNMTs is necessary for further SCNT studies. In the present study, we examined the potential use of RNA interference for knockdown of DNMT mRNA in bovine fibroblast cells that were commonly used as karyoplast donors in SCNT studies. We designed three siRNAs corresponding to DNMT1, DNMT2 and DNMT3a mRNA. In Experiment 1, to optimize transfection conditions, fluorescence and cell viability after transfection were evaluated at different concentrations of transfection reagent using a FITC-labeled nonsilencing control siRNA. Although fluorescence was observed in all groups transfected except for the negative control group, transfection with a higher concentration of transfection reagent significantly decreased in cell viability (P<0.05). In Experiment 2, the amount of DNMT mRNA was measured by real-time PCR at 0, 48 and 96 h after siRNA transfection into the cells. The levels of each DNMT mRNA were significantly decreased at 48 and 96 h after transfection (P<0.01). Furthermore, decreased expression of DNMT1 protein was confirmed by western blotting. In Experiment 3, the DNA methylation statuses were analyzed in each of the siRNA-transfected groups. The DNMT1 siRNA-transfected group had a significantly decreased level of DNA methylation (P<0.05), but the other groups did not. Our data demonstrate that RNA interference with siRNA can be analyzed the function and expression of DNMT genes in bovine fibroblast cells. The present study provides useful information for further SCNT studies. PMID- 19815987 TI - Characterization of maternal antigen that embryos require (MATER/NLRP5) gene and protein in pig somatic tissues and germ cells. AB - Maternal effect genes produce mRNA or proteins that accumulate in the egg during oogenesis and control the developmental program until embryonic genome activation takes place. NLRP5 (NLR family, Pyrin domain containing 5), also called MATER (Maternal Antigen That Embryos Require) is one of the genes required for normal early embryonic development, although its precise function remains to be elucidated. The aim of the present study was to analyze the NLRP5 gene expression pattern and protein distribution in somatic tissues and germ cells in the pig. Reverse transcription was performed on mRNA from germinal vescicle (GV) oocytes and total RNA from spermatozoa and tissues from different organs. The transcript for NLRP5 gene was identified only in ovaries and oocytes. The presence of NLRP5 protein was detected only in ovaries by western blot analysis and immunohistochemistry. PMID- 19815988 TI - Hyperactivated motility of frozen-thawed spermatozoa from fertile and subfertile Japanese black bulls induced by cyclic adenosine 3',5'-monophosphate analogue, cBiMPS. AB - This study investigated whether a cyclic adenosine 3',5'-monophosphate (cAMP) analogue, cBiMPS, could induce hyperactivated motility in frozen-thawed Japanese Black bull spermatozoa and compared the ability of spermatozoa to undergo hyperactivation between fertile and subfertile bulls. Frozen-thawed spermatozoa from 3 fertile and 2 subfertile bulls were washed, suspended in BO-Hepes medium and incubated in the presence of 0.1 mM cBiMPS for up to 4 h. At 1-h intervals, the spermatozoa were examined for hyperactivated motility. The proportions of spermatozoa showing a circular swimming pattern with asymmetrical flagellar beating and those showing whiplash beating of flagella to the total number of motile spermatozoa were expressed as C% and W%, respectively. The motile spermatozoa % was barely affected by treatment with cBiMPS or the fertility status of the sperm donor, although it gradually decreased in all sperm samples during the 4-h incubation. In the fertile bulls, the C% was 0% at 0 h of incubation but rapidly increased during the 1-h incubation with cBiMPS. It then decreased slightly towards 4 h concomitantly with a gradual increase in W% towards 4 h. In the subfertile bulls, however, the cBiMPS-induced increase of C% was delayed for 1-3 h, although the incubation time-related changes in mean W% were similar between the fertile and subfertile bulls. In the vehicle controls for cBiMPS, the C% and W% were 0% throughout incubation for all the samples examined. The results suggest that hyperactivation of the flagellum can be induced by the cAMP analogue, cBiMPS, in frozen-thawed Japanese Black bull spermatozoa and that induction of hyperactivation may serve as a useful tool for detection of functional abnormality of spermatozoa from subfertile Japanese Black bulls. PMID- 19815989 TI - Effects of season and reproductive phase on the quality, quantity and developmental competence of oocytes aspirated from Japanese black cows. AB - The aim of the present study was to determine whether the season (hot and cool) and reproductive phase (pregnant and non-pregnant) of the cow affect follicular recruitment and oocyte development. Follicular oocytes were aspirated from Japanese black cows by the ovum pick-up (OPU) method, which was performed 2 to 6 times within 1.5 months in pregnant cows and 2 to 4 times within 2 months in non pregnant cows, during the hot (July to September) and cool (October to November) seasons. After follicular aspiration, the number and morphology of cumulus-oocyte complexes (COCs) and the developmental competence of oocytes after in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture were evaluated. The quality of aspirated COCs did not differ between the hot and cool seasons, irrespective of the reproductive phase of the donor cows. In the pregnant cows, the season did not affect follicular recruitment, early embryonic development or the quality of embryos. In the non-pregnant cows, however, the mean number of aspirated follicles and collected oocytes decreased during the hot season as compared with the cool season. When the data for the 2 seasons were combined to assess the effects of reproductive phase on oocyte development, the total proportions of cleavage, development into blastocysts and freezable embryos were higher for embryos obtained from pregnant cows (P<0.05) than those obtained from non-pregnant cows. In conclusion, the season did not have any apparent effects on the quality of aspirated COCs and the developmental competence of oocytes after IVM-IVF, but it may affect follicular recruitment in non-pregnant cows. Moreover, the reproductive phase may influence the developmental competence of the recovered oocytes. PMID- 19815990 TI - Measurements of the calibration factor of a light scattering dust monitor for CO(2) arc welding fumes--the effect of the sampling location on the calibration factor. PMID- 19815991 TI - [Health impact assessment of occupational health policy reform at a multinational chemical company in Japan]. AB - A health impact assessment (HIA) was conducted to identify potential health impacts arising from policy reform of occupational health and safety at S chemical company, a multinational global company that employs about 13,000 workers. A multidisciplinary team of health professionals including occupational physicians, an epidemiologist, and public health researchers oversaw the HIA. A project manager from S-company was also involved in the whole HIA process. A literature review, profiling using annual health examination data and interviews with stakeholders and key informants were undertaken in order to identify possible impacts. A range of positive and negative health impacts were identified and develop recommendations for implementation of the new occupational health policy were proposed. The HIA added value to the planning process for the occupational health policy reform. PMID- 19815992 TI - Magnetic resonance signal intensity from retrodiscal tissue related to joint effusion status and disc displacement in elderly patients with temporomandibular joint disorders. AB - Magnetic resonance imaging (MRI) on patients with temporomandibular joint disorders (TMD) has revealed that a decrease and/or increase in signal intensity from retrodiscal tissue, joint effusion (the excessive accumulation of joint fluid) and articular disc displacement are related to TMD. However, the effect of aging on these phenomena has yet to be clarified. This study was carried out to explore the relationship between changes in signal intensity from retrodiscal tissue, joint fluid status and pathological disc conditions in elderly patients with TMD. Twenty patients aged over 60 years were examined. They consisted of one man and 19 women, and ranged between 60 and 79 years in age (mean, 66.0 years). The relationships between decreased signal intensity on proton-density-weighted (PDW) images and increased signal intensity on T2-weighted (T2W) MR images from retrodiscal tissue, joint fluid status and state of articular disc were examined. Joint fluid status was classified into 5 levels by extent of high signal areas in upper and lower articular spaces on T2W images. Disc displacement status was evaluated by PDW images. The Wilcoxon test was applied for the statistical analysis. The group showing increased T2W signal intensities from the retrodiscal tissue consisted of 31 out of 40 joints (77.5%). This group showed a significant difference in comparison with the other groups in which no apparent joint fluid was shown (p<0.05). There were no statistically significant differences among other categories. The results suggest a negative relationship between joint fluid and increased signal intensity from retrodiscal tissue due to reflection of the inflammatory reaction in TM joints. PMID- 19815993 TI - Geographic distribution of postgraduate dental trainees in Japan. AB - Postgraduate clinical training for dentists in Japan became mandatory in April 2006. Mandatory postgraduate clinical training for physicians has been criticized as having accelerated the imbalance in distribution of physicians. This suggests the danger that the same phenomenon might occur in distribution of dentists. It is also necessary to investigate the geographic distribution of dental trainees and practicing dentists in Japan. In this study, the number of dental trainees enrolled in each clinical training program and number that had actually received clinical training at each facility were compared by prefecture. The results suggest that disparities in the number of dental trainees among prefectures are being compensated for by movement across prefectural borders under the clinical training facilities-group system. Postgraduate dental trainees, however, showed a significantly greater imbalance in geographic distribution than practicing dentists. Continuation of the postgraduate clinical training for dentists under the existing system may accelerate this imbalance in distribution of dentists. To prevent this, practical measures should be taken in accordance with the coming review of the system, based on research regarding changes in geographic distribution of dental trainees. PMID- 19815994 TI - Influence of mandibular fixation method on stability of the maxillary occlusal plane after occlusal plane alteration. AB - In this study, we investigated how method of mandibular fixation influenced longterm postoperative stability of the maxilla in Class III cases. In particular, we investigated change in the maxillary occlusal plane after Occlusal Plane Alteration. Therefore, we focused on change in the palatal plane to evaluate stability of the maxillary occlusal plane, as the position of the palatal plane affects the maxillary occlusal plane. This study included 16 patients diagnosed with mandibular protrusion. Alteration of the occlusal plane was achieved by clockwise rotation of the maxilla by Le Fort I osteotomy and mandibular setback was performed by bilateral sagittal split ramus osteotomy. We analyzed and examined lateral cephalometric radiographs taken at 1 month, 3 months, 6 months, and 1 year after surgery. Stability achieved by two methods of mandibular fixation was compared. In one group of patients (group S) titanium screws were used, and in the other group (group P) titanium-locking mini-plates were used. No significant displacement was recognized in group S, whereas an approximately 0.7mm upward vertical displacement was recognized in the anterior nasal spine in group P. As a result, not only the angle of the palatal plane and S-N plane, but also occlusal plane angle in group P showed a greater decrease than that in group S. The results suggest that fixing the mandible with screws yielded greater stability of the maxilla and maxillary occlusal plane than fixing the mandible with titanium plates. PMID- 19815995 TI - Relationship between oral tissue blood flow and oxygen tension in rabbit. AB - Several studies have reported tissue blood flow and tissue oxygen tension during anesthesia, whereas there are few reports that discuss the relationship between tissue blood flow and tissue oxygen tension. The goal of this study was to investigate the relationship between these two variables before and after stellate ganglion block (SGB). We utilized 8 male Japan White rabbits. Anesthesia was maintained with propofol. For SGB, 0.2 ml of 1% lidocaine was injected and changes in mandibular bone marrow blood flow (BBF) and mandibular bone marrow oxygen tension (PbO(2)) were observed (Group B). After the observed variables completely recovered, 0.2 ml of 1% lidocaine was again injected and changes in masseter muscle blood flow (MBF) and masseter muscle oxygen tension (PmO(2)) were observed (Group M). The observed variables were hemodynamic parameters, common carotid artery blood flow, L (left side)-BBF, R (right side)-BBF, L-PbO(2), R PbO(2), L-MBF, R-MBF, L-PmO(2) and R-PmO(2). In both groups, positive correlations (r=0.99; y=0.68x - 3.49 in Group B and r=0.99; y=0.62x + 0.47 in Group M) were observed when tissue blood flow was set at the X-axis and tissue oxygen tension at the Y-axis. In contrast, when tissue blood flow was set at the Y-axis and tissue oxygen tension at the X-axis, two regression lines almost overlapped (y=1.47x + 5.12 in Group B and y=1.59x - 0.28 in Group M). Therefore, it is estimated that tissue blood flow increases by approximately 15 ml/min/100g when tissue oxygen tension increases by 10 mmHg. In conclusion, there is a positive correlation between tissue blood flow and tissue oxygen tension. Changes in tissue oxygen tension should reflect the increase or decrease in tissue blood flow. PMID- 19815996 TI - Reestablishment of occlusion with prosthesis and composite resin restorations. AB - Here, we present a case report on prosthetic reconstruction of posterior teeth and composite resin restoration of anterior teeth yielding considerable esthetic improvement, reestablishment of disocclusion guides and function. PMID- 19815997 TI - Root coverage technique with enamel matrix derivative. AB - Various periodontal plastic surgical techniques are employed in obtaining root coverage. Recently, the use of an enamel matrix derivative (EMD) has been reported in such treatment. We report 2 cases of root coverage surgery with a coronally positioned flap in combination with EMD (CPF+EMD) and connective tissue graft in combination with EMD (CTG+EMD). Case 1: The patient was a 25-year-old woman referred to Suidobashi Hospital, Tokyo Dental College for root coverage surgery on the lower right first premolar. Gingival recession was classified as Miller Class II, as no alveolar bone loss or loss of attachment was observed in the interdental area, although recession had progressed to the mucogingival junction. The patient was diagnosed with local gingival recession caused by excessive tooth brushing. Primary conservative treatment failed to reduce the gingival recession. Subsequently, root coverage surgery with CPF+EMD was carried out. As observation at the 1-year follow-up revealed complete root coverage and no recurrence of root exposure or subjective symptoms, the postoperative course was considered to be favorable. Case 2: The patient was a 39-year-old woman referred to Suidobashi Hospital, Tokyo Dental College for root coverage surgery on the lower left canine. Gingival recession was classified as Miller Class II. Root coverage surgery with CTG+EMD was carried out. As observation at the 2-month follow-up revealed complete root coverage and no recurrence of root exposure, the postoperative course was considered to be favorable. These 2 cases indicate the effectiveness of root coverage surgery with CPF+EMD and CTG+EMD. PMID- 19815998 TI - Risperidone treatment increases CB1 receptor binding in rat brain. AB - BACKGROUND/AIMS: Body weight gain is a common side effect of treatment with antipsychotics, but the mechanisms underlying this weight gain are unknown. Several factors may be involved in antipsychotic-induced body weight gain including the cannabinoid receptor 1 (CB(1)), the serotonin receptor 2C, the ghrelin receptor, neuropeptide Y, adiponectin and proopiomelanocortin. We investigated whether the expression of these factors was affected in rats chronically treated with the antipsychotic risperidone. METHODS: Male Sprague Dawley rats were treated with risperidone (1.0 mg/kg/day) or vehicle (20% hydroxypropyl beta-cyclodextrin) for 28 days. Expression of the aforementioned factors were examined together with plasma prolactin and ghrelin levels. RESULTS: No difference in body weight gained during treatment was observed between risperidone and vehicle treated rats, but plasma risperidone levels positively correlated with visceral fat mass. Risperidone treatment increased CB(1) receptor binding in the arcuate nucleus (40%), hippocampus (25-30%) and amygdala (35%) without concurrent alterations in the CB(1) receptor mRNA. Risperidone treatment increased adiponectin mRNA. CONCLUSION: The present study showed that risperidone treatment altered CB(1) receptor binding in the rat brain. Risperidone-induced adiposity and metabolic dysfunction in the clinic may be explained by increased CB(1) receptor density in brain regions involved in appetite and regulation of metabolic function. PMID- 19815999 TI - Pharmacogenomics: role in medicines approval and clinical use. AB - Pharmacogenomics (PGx) and pharmacogenetics (PGt) are emerging interdisciplinary areas recently defined by the regulatory authorities at an international level as 'the investigation of variations of DNA and RNA characteristics as related to drug response' (PGx), and the study of 'the influence of variations in DNA sequence on drug efficacy and toxicity' (PGt). In recent years a number of studies have in fact produced growing evidence that, besides the effects of age, sex, diseases, and different drugs interactions, genetic factors play a role in the inter-individual variability of drugs response. The increasing genomic knowledge has also raised the profile and role of the so called 'genomic biomarkers' (GBs) in drug development, approval, and clinical use. The aims of this review are to (a) revisit the general understanding of the role of genomics and genetics in drug response, (b) provide an update on the definition and classification criteria of GBs as recently defined at a global level by regulatory agencies such as the European Medicines Agency and the Food and Drug Administration, and (c) illustrate with some examples current and potential applications of biomarkers in clinical practice and in drug development. PMID- 19816000 TI - Acute and chronic effects of smoking on inflammation markers in exhaled breath condensate in current smokers. AB - BACKGROUND: Long-term cigarette smoking is associated with pulmonary inflammation, but the acute effects of smoking have been less well studied. Analysis of the exhaled breath condensate (EBC) can provide noninvasive markers that might be indicative of inflammation. OBJECTIVES: The aim of the study was to determine whether the pH , electrical conductivity and the levels of ammonium and interleukin 8 (IL-8) of EBC were altered in smokers and whether they changed after smoking a single cigarette. METHODS: We included 19 healthy nonsmokers (controls), 29 asymptomatic smokers, 10 patients with stable chronic obstructive pulmonary disease (COPD) [Global Initiative for Chronic Obstructive Lung Disease stages (GOLD) stages II-III], and 10 patients with exacerbated COPD. In 13 smokers, EBC was also analyzed before and after smoking. EBC was obtained during 10 min tidal breathing with a cooled RTube. pH was determined after deaeration with argon. RESULTS: Acute smoking did not alter the pH or ammonium and IL-8 levels, but raised conductivity. As in COPD patients, the pH was significantly decreased in chronic smokers with a history of at least 10 pack-years compared to controls. CONCLUSIONS: EBC can be used to detect the acute and chronic effects of smoking. The increased conductivity of EBC after smoking suggests acute inflammatory effects. The reduced pH in chronic smokers shows cigarette-induced inflammation. PMID- 19816002 TI - Acute oligohydramnios: antenatal expression of VURD syndrome? AB - OBJECTIVE: Oligohydramnios (OA) is nowadays regarded as one of the best markers of renal function (RF) impairment in bladder outlet obstruction (BOO) detected in utero. As such, its onset is usually early and progressive because of decline in fetal urine production. A series of acute OA complicating pregnancies with BOO has never been reported. METHODS: Over a 7-year period, 5 fetuses with in utero suspicion of BOO exhibited an abrupt decrease of amniotic fluid after the 30th week of gestation. RESULTS: All fetuses were delivered by cesarean section: diagnosis was posterior urethral valves in 3 cases, urethral atresia in 1, and prune-belly syndrome in 1. Urologic work-up demonstrated a unilateral vesicoureteral reflux dysplasia (VURD syndrome) in all 5 fetuses. RF at 1 year was normal in 4 fetuses and impaired in 1. CONCLUSIONS: Besides obstetrical reasons, OA may also have acute onset occurring in the presence of anomalies of the urinary tract; although diagnosis is almost always BOO, functional and anatomical characteristics of the urinary tract are those of VURD syndrome with a non-functioning, refluxing renal unit. The associated acute OA/VURD syndrome may represent a milder expression of a pop-off mechanism advocated in this syndrome with a more favorable prognosis than progressive OA detected early in pregnancy. PMID- 19816001 TI - Angiogenic and angiostatic chemokines in idiopathic pulmonary fibrosis and granulomatous lung disease. AB - BACKGROUND: Angiogenesis-angiostasis balance and leukocyte recruitment are influenced by different concentrations of distinct chemokines. OBJECTIVE: To investigate the relative contribution of angiogenic and angiostatic CXC chemokines to the pathogenesis of idiopathic pulmonary fibrosis (IPF) and granulomatous lung diseases, we examined the in vitro production of an angiogenic chemokine (IL-8), and 2 angiostatic chemokines (IP-10 and MIG) by alveolar macrophages. METHODS: Alveolar macrophages from 16 patients with granulomatous lung diseases [8 with sarcoidosis, 8 with extrinsic allergic alveolitis (EAA)], 16 patients with IPF, and 8 control subjects were cultured for 24 h. IL-8, IL-18, IP-10 and MIG in the culture supernatants were measured by a fluorescent bead based multiplex technique. RESULTS: In IPF patients, IL-8 was increased and correlated with bronchoalveolar lavage (BAL) neutrophils, whereas the levels of IP-10 and MIG were normal. In sarcoidosis and EAA patients, IL-8, IP-10, and MIG were all increased and IP-10 and MIG correlated with IL-18, a Th1 cytokine, and the percentage and number of BAL lymphocytes. CONCLUSIONS: The difference in the expression of CXC chemokines and a Th1 cytokine may contribute to the different immunopathogenesis, clinical course and responsiveness to treatment of these diseases. PMID- 19816004 TI - Assessment and reproducibility of non-eosinophilic asthma using induced sputum. AB - BACKGROUND: The assessment of eosinophilic airway inflammation using induced sputum identifies a corticosteroid-responsive subtype that can be used to guide anti-inflammatory therapy. The stability of airway inflammation in asthma over time is not known, yet this information is crucial to inflammation-based patient management. OBJECTIVE: The aim of this study was to determine the reproducibility of non-eosinophilic asthma. METHODS: Participants with stable asthma (n = 26) underwent a sputum induction each month for 5 months. Sputum was dispersed and differential cell counts were performed. The reproducibility of inflammatory subtype with different eosinophil cut points (starting at 1% eosinophils) was examined and the minimum number of visits required to determine inflammatory subtype was calculated. RESULTS: One hundred and twenty- two sputum samples were obtained (success 94%). All cut points greater than 2% eosinophils were reproducible and a 3% cut point resulted in the highest agreement with a kappa statistic of 0.538. Specificity and sensitivity were high for determining inflammatory subtype after 1 or 3 sputum samples. CONCLUSIONS: A cut point of 3% eosinophils should be used to distinguish eosinophilic from non-eosinophilic airway inflammation in asthma. This allows a single sputum sample to be used to reliably determine the presence of eosinophilia. PMID- 19816003 TI - Evidence for a founder mutation in the cathepsin C gene in three families with Papillon-Lefevre syndrome. AB - BACKGROUND: Papillon-Lefevre syndrome (PLS; OMIM 245000) is a rare autosomal recessive disorder. Clinically, PLS is characterized by hyperkeratosis involving the palms, soles, elbows and knees which is followed later on by periodontitis, destruction of alveolar bone and loss of primary and permanent teeth. The condition is caused by mutations in the cathepsin C (CTSC) gene. METHODS: We analyzed the DNA of members from 3 consanguineous families for mutations in the CTSC gene by direct sequencing analysis. We then performed haplotype analysis. RESULTS: We identified an identical recurrent missense mutation, R272P, in all 3 families. Microsatellite marker analysis around the CTSC gene revealed the same haplotype on the mutation-carrying allele in all 3 families. CONCLUSION: The presence of this common mutation in families from 2 different geographical areas provides evidence for a founder effect for CTSC mutations in PLS. PMID- 19816005 TI - Vascular endothelial growth factors, angiogenesis, and survival in human ileal enterochromaffin cell carcinoids. AB - BACKGROUND AND AIMS: Well-differentiated neuro-endocrine ileal carcinoids are composed of serotonin-producing enterochromaffin (EC) cells. Life expectancy is determined by metastatic spread to the liver because medical treatment options are still very limited. Selective inhibition of angiogenesis or lymphangiogenesis might prevent tumour growth and metastatic spread. We examined the role of the vascular endothelial growth factors (VEGFs) A, B, C, D, and their receptors (VEGFRs) 1, 2, 3 in angiogenesis and lymphangiogenesis of ileal EC cell carcinoids with and without liver metastases. METHODS: The expression of various VEGFs and VEGFRs was determined by quantitative real-time RT-PCR in healthy mucosa, primary tumour, lymph node metastases and liver metastases of 25 patients with ileal EC cell carcinoids. Microvessel density (MVD) was determined by CD-31 staining in primary tumours and lymphatic vessel density (LVD) by LYVE-1 staining. VEGF expression levels, MVD, LVD, and patients' survival time were correlated using logistic regression and Kaplan-Meier survival analysis. RESULTS: VEGF-A was highly expressed with no difference between normal mucosa and tumours. VEGF-B and -D as well as VEGFR-1 and -2 expression levels were significantly increased in the tumours when compared to normal mucosa. Patients with liver metastasis, however, had a significantly lower expression of the factors A, B, and C and the receptors 2 and 3. MVD in primary tumours positively correlated with the expression of VEGF ligands and their receptors, except for VEGF-D. LVD did not correlate with any VEGF ligand or receptor. Interestingly, low expression levels of VEGF-B were associated with poor survival. CONCLUSION: Patients with more aggressive metastatic spreading had relatively decreased expression levels of VEGF ligands and receptors. Thus, anti-angiogenic therapy may not be a suitable target in metastatic ileal EC cell carcinoids. PMID- 19816006 TI - Absence of the V617F JAK2 mutation in the lymphoid compartment in a patient with essential thrombocythemia and B-chronic lymphocytic leukemia and in two relatives with lymphoproliferative disorders. AB - BACKGROUND: Myeloproliferative neoplasms likely involve both myeloid and lymphoid lineages. Nevertheless, the coincidence of chronic myeloproliferative and lymphoproliferative diseases in the same patient is a rare phenomenon. METHODS: We report a case of a patient having essential thrombocythemia (ET) and B-chronic lymphocytic leukemia (B-CLL). In this patient and in 2 relatives with lymphoproliferative disorders, we searched for JAK2(V617F) mutation in lymphocytes. RESULTS: In the patient with ET and B-CLL, we identified homozygous JAK2(V617F) mutation in the granulocytic compartment. Both relatives were heterozygous for JAK2(V617F) mutation, whereas no mutation signal could be detected in the lymphoid compartment of all 3 patients. CONCLUSION: Our results seem to confirm that CLL cases are negative for JAK2(V617F) mutation in B- and T lymphocyte populations.Presence of JAK2(V617F) mutation in subjects without myeloproliferative diseases could indicate an increased risk of a future myeloproliferative neoplasm development. PMID- 19816007 TI - Prenatal diagnosis of hemoglobin Bart's hydrops fetalis with gap-PCR system. PMID- 19816009 TI - Arsenic trioxide in a hemodialytic patient with acute promyelocytic leukemia. PMID- 19816011 TI - Self-monitoring of oral anticoagulation therapy in children. AB - This study aimed to investigate the accuracy of home International Normalized Ratio (INR) self-monitoring in pediatric patients on long-term oral anticoagulation therapy. Statistical and clinical agreement of INR values from capillary whole blood samples measured by 2 different portable prothrombin time monitors (CoaguChek S and XS) and venous blood samples measured by a laboratory coagulation analyzer were evaluated using the Bland-Altman analysis. Eighty-three INR comparisons (56 using the CoaguChek S and 27 using the CoaguChek XS) were obtained from 35 children aged 4 months to 18 years. Mean differences between venous and capillary INR values and their limits of agreement were -0.04 (-0.63 to 0.55) overall, 0.006 (-0.63 to 0.65) for the CoaguChek S and -0.13 (-0.57 to 0.31) for the CoaguChek XS. The Pearson correlation coefficients were 0.88 overall, 0.84 for the CoaguChek S and 0.95 for the CoaguChek XS. Expanded and narrow agreements for all patients were 97.6 and 94%, respectively. In conclusion, home INR self-monitoring is accurate for children requiring long-term oral anticoagulation therapy. Our data suggest that INR self-monitoring with the newer CoaguChek XS is more accurate than with the older CoaguChek S monitor. PMID- 19816010 TI - Effect of L-asparaginase combined with vincristine and prednisolone on acute myeloblastic leukemia (M0) associated with non-Hodgkin lymphoma. AB - A 66-year-old Japanese woman was referred to us because of severe anemia and fever and presented at our hospital. She was eventually diagnosed as having acute myeloblastic leukemia (AML; M0) with non-Hodgkin lymphoma (NHL). We investigated the therapeutic efficacy of L-asparaginase (L-Asp), vincristine and prednisolone for both her AML and NHL. Asparagine synthetase (AS) activity in her AML blast cells was undetectable. A lymph node biopsy specimen revealed NHL of the marginal zone B cell type. Complete remission (CR) of AML and NHL was achieved. CR of the AML lasted for 18 months without further consolidation therapy. We conclude that L-Asp can be an effective drug for the treatment of AML in which blasts are negative for AS. PMID- 19816012 TI - Elevated circulating stromal-derived factor-1 levels in sickle cell disease. AB - Inflammation and angiogenesis are of importance in the pathophysiology of sickle cell disease (SCD). Recently, the chemokine stromal-derived factor-1 (SDF-1) has been shown to be a key mediator of angiogenesis and inflammation. In this study we determined serum SDF-1 levels in consecutive adult sickle cell patients during the clinically asymptomatic state as well as during painful crisis. Serum SDF-1 levels were significantly elevated in HbSS/HbSbeta(0)-thalassaemia patients [n = 42; 5,177 pg/ml (2,438-7,246)] compared to HbSC/HbSbeta(+)-thalassaemia patients [n = 16; 2,405 pg/ml (1,365-3,047)] and healthy HbAA controls [n = 45; 2,894 pg/ml (2,577-3,334)] (p = 0.001). No significant increments were observed during painful crisis (n = 40). SDF-1 levels were significantly higher in SCD patients with pulmonary hypertension (PHT) compared to patients without PHT. Elevated circulating SDF-1 levels occur in patients with SCD and may play a role in the pathophysiology of SCD-related PHT. PMID- 19816013 TI - Mutagenicity of immunosuppressive medications among renal transplant recipients. AB - BACKGROUND: Immunosuppressive therapy is essential for patients after renal transplantation, but it may have mutagenic side effects. The frequency of micronuclei (MN) assessed by the cytokinesis block assay is well established in the diagnosis of DNA damage. METHODS: We examined 79 patients before and after renal transplantation with the cytokinesis block assay. For MN evaluation, the criteria of the Human Micron Nucleus (HUMN) project were used. RESULTS: Age and sex had no influence on the number of MN before transplantation. Patients with a shorter time on dialysis had fewer MN than patients with a longer time on dialysis. After 3 weeks of immunosuppressive therapy, the ability of cells to proliferate was reduced; therefore, only 36 patients could be analyzed. In these patients, MN frequency rose significantly. There was no linear relationship between MN after transplantation and age, sex, time on dialysis or graft function. The majority of patients (79%) were treated with cyclosporine A, mycophenolate mofetil and methylprednisolone when leaving the hospital. There was no difference between the different therapeutic schemes with regard to MN frequency after transplantation, but patients with mycophenolate mofetil showed less cellular proliferation. The function of the transplanted organ or the occurrence of rejection had no effect on MN frequency after transplantation. CONCLUSION: In patients with renal transplantation, immunosuppressive therapy intensifies the genotoxic damage of the preceding chronic renal failure. PMID- 19816014 TI - Erythropoiesis stimulatory agent- resistant anemia in dialysis patients: review of causes and management. AB - Despite new therapeutic options and treatment strategies, anemia still remains one of the major complications of chronic kidney disease (CKD), especially in patients undergoing chronic hemodialysis for end-stage renal disease. Successful management of anemia is a central part of patient care that may improve clinical outcomes. Although the National Kidney Foundation Dialysis Outcomes Quality Initiative (NKF-DOQI) working group reformulated its recommendations by stating that the hemoglobin target in patients receiving erythropoiesis stimulatory agents (ESA) should generally be 11-12 g/dl, this target value can not be achieved in many of them, despite treatment with high doses of ESA. The aim of the present review is to provide an update of the recent literature on causes and possible management of ESA-resistant anemia in CKD patients. PMID- 19816015 TI - Coronary calcifications in end-stage renal disease patients: a new link between osteoprotegerin, diabetes and body mass index? AB - The aim of the study was to assess the factors potentially involved in coronary artery calcifications (CAC) in end-stage renal disease patients. 253 hemodialysis (HD) patients (92 females, 161 males), aged 62.5 +/- 13.5, who had been on HD treatment for at least 6 months, were enrolled in a cross-sectional study. Calcium-phosphate product (Ca x P), body mass index (BMI), fetuin-A, osteoprotegerin (OPG), osteopontin, transforming growth factor-beta1 (TGF-beta1), fibroblast growth factor-23 (FGF-23) and matrix Gla protein (MGP) were considered. CAC was assessed using multislice spiral computed tomography and calcium score was quantified by means of the Agatston score. The median calcium score was 364 Agatston (range 0-7,336). CAC was detected in 228/253 patients (90.1%). Multivariate regression analysis, adjusted for age and for dialysis vintage, showed that TGF-beta1, OPG and days with Ca x P >55 mg/dl are independent predictors of CAC, while MGP was shown to be a protective factor. Surprisingly, results showed that BMI was a protective factor too: the interpolation with cubic spline function revealed a significant reduction in calcium score in patients with a high BMI (>28). However, when diabetes was considered in the regression analysis, only OPG emerged as a predictor of a high CAC score. The interpolation with spline function continued to show a significant reduction in CAC score in nondiabetic and in diabetic patients with the highest BMI quartile. The protective effect of a high BMI on CAC might represent another example of inverse biology in dialysis patients but it needs to be further addressed in larger longitudinal studies. PMID- 19816016 TI - Extracorporeal membrane oxygenation and high-dose continuous veno-venous hemodiafiltration in a young child as a successful bridge to heart transplant for management of combined heart and kidney failure: a case report. AB - Combined heart and kidney transplant is an established treatment modality in patients with coexisting end-stage heart and kidney failure. However, there is scarce information on the optimal management of children that are listed for cardiac transplantation with mild to severe renal dysfunction. Herein we report the case of a young child who presented with life-threatening dilative cardiomyopathy and severe renal dysfunction, and who required urgent extracorporeal membrane oxygenation associated with continuous veno-venous hemodiafiltration and eventually underwent successful isolated heart transplantation followed by complete recovery of kidney function. PMID- 19816017 TI - Mathematical analysis of the long-term efficacy of daily home hemodialysis therapy with a cold dialysate regeneration system. AB - BACKGROUND/AIM: We previously developed the cold dialysate regeneration system (CDRS) for use in daily home hemodialysis (HD). This study evaluates the long term efficacy of the CDRS using a mathematical method. METHOD: HD with the CDRS was simulated using a method that integrates the mass-transfer model and the dialyzer solute kinetics model with a newly proposed model of CDRS function. RESULTS: We mathematically assessed the long-term efficacy of HD with the CDRS. The weekly treatment time required for HD with the CDRS was reduced significantly using the daily dialysis method; it required only an 11% longer treatment time to obtain the corrected equivalent renal clearance and a 14% shorter time for the standard Kt/V, compared with thrice weekly treatment with conventional HD. CONCLUSION: By developing a mathematical model to test the long-term efficacy of the new CDRS, we showed that HD with the CDRS is an efficient means of daily home hemodialysis therapy. PMID- 19816018 TI - A double-blind, placebo-controlled, randomized phase II pilot study to investigate the potential efficacy of the traditional chinese medicine Neuroaid (MLC 601) in enhancing recovery after stroke (TIERS). AB - BACKGROUND AND OBJECTIVE: Previous clinical studies have shown that Neuroaid (MLC 601) may be beneficial in post-stroke rehabilitation. Our aim was to investigate the efficacy of Neuroaid on motor recovery in ischemic stroke patients using rehabilitation endpoints in accordance with the International Conference on Harmonization/Good Clinical Practice guidelines, in order to provide predictive information for further larger trials. METHODS: This is a phase II double-blind, placebo-controlled pilot study of 40 subjects admitted with a recent (less than 1 month) ischemic stroke. All subjects were given either Neuroaid or placebo, 4 capsules 3 times a day for 4 weeks. Fugl-Meyer Assessment (FMA), National Institutes of Health Stroke Scale and Functional Independence Measure scores were measured at initiation of the treatment, and at 4 and 8 weeks. RESULTS: None of the outcomes was statistically significant between the two groups. However, FMA scores showed a positive trend for improvement with Neuroaid treatment over time. Subgroup analysis of subjects with posterior circulation infarction and severe stroke both showed a tendency for better recovery. CONCLUSION: Some positive trends were observed in the Neuroaid group. A larger multicenter trial focusing on severe stroke patients is needed to better evaluate the role of Neuroaid in aiding stroke recovery in rehabilitation. PMID- 19816019 TI - Prediction of 30-day mortality in older patients with a first acute myocardial infarction. AB - OBJECTIVES: This study sought predictors of mortality in patients aged >or=75 years with a first ST-segment elevation myocardial infarction (STEMI) and evaluated the validity of the GUSTO-I and TIMI risk models. METHODS: Clinical variables, treatment and mortality data from 433 consecutive patients were collected. Univariable and multivariable logistic regression analyses were applied to identify baseline factors associated with 30-day mortality. Subsequently a model predicting 30-day mortality was created and compared with the performance of the GUSTO-I and TIMI models. RESULTS: After adjustment, a higher Killip class was the most important predictor (OR 16.1; 95% CI 5.7-45.6). Elevated heart rate, longer time delay to admission, hyperglycemia and older age were also associated with increased risk. Patients with hypercholesterolemia had a significantly lower risk (OR 0.46; 95% CI 0.24-0.86). Discrimination (c statistic 0.79, 95% CI 0.75-0.84) and calibration (Hosmer-Lemeshow 6, p = 0.5) of our model were good. The GUSTO-I and TIMI risk scores produced adequate discrimination within our dataset (c-statistic 0.76, 95% CI 0.71-0.81, and c statistic 0.77, 95% CI 0.72-0.82, respectively), but calibration was not satisfactory (HL 21.8, p = 0.005 for GUSTO-I, and HL 20.6, p = 0.008 for TIMI). CONCLUSIONS: Short-term mortality in elderly patients with a first STEMI depends most importantly on initial clinical and hemodynamic status. The GUSTO-I and TIMI models are insufficiently adequate for providing an exact estimate of 30-day mortality risk. PMID- 19816020 TI - Management of spontaneous coronary artery dissection: review of the literature and discussion based on a series of 12 young women with acute coronary syndrome. AB - Spontaneous coronary artery dissection (SCAD) is a rare pathology, principally affecting young women free of atheroma risk factors. Its physiopathology remains little understood, and the prognosis for such acute coronary syndromes is poor, as they occur suddenly. Management is often difficult, and no guidelines exist. The present single-center retrospective study concerns 12 cases of SCAD occurring between 2001 and 2008 in female patients under the age of 60. Eleven patients survived, with a favorable long-term evolution. Only 2 had conservative medical therapy, the other 10 undergoing percutaneous coronary intervention (2 procedures involving a coronary artery bypass graft). On the basis of this series and data from the literature, we suggest a strategy to improve the often dire prognosis of SCAD. Emergency angiography to confirm diagnosis is essential. Treatment should be guided by the extent of the lesions, the myocardial ischemia and the hemodynamic status. Conservative medical therapy is a reasonable approach in the case of distal dissection or conserved coronary flow. Percutaneous coronary intervention is feasible in the acute phase to restore coronary perfusion and hemodynamic stability. Surgery - emergency bypass or assisted circulation - should be restricted to cases where percutaneous coronary intervention has failed or is impossible. PMID- 19816021 TI - Laparoscopic Heller-Dor surgery for esophageal achalasia: impact of intraoperative real-time manometric feedback on postoperative outcomes. AB - BACKGROUND: Laparoscopic Heller myotomy with Dor fundoplication (LHD) is one of the most established surgical procedures for esophageal achalasia. Preoperative esophageal manometry has been reported as useful to evaluate lower esophageal sphincter (LES) pressure. However, the feasibility, safety, and impact of its intraoperative use have not been fully evaluated, especially when enhanced with real-time 3-D pressure imaging. METHODS: LHD was attempted on 24 consecutive patients with esophageal achalasia. Manometry was performed at 3 time points during LHD: before myotomy, after myotomy, and after fundoplication. Investigations included esophagography, manometry, and 24-hour esophageal pH monitoring in the preoperative, short-term (0-5 months) and long-term (1-3 years) follow-up periods. RESULTS: The 3-D intraoperative manometric images were presented to the surgical crew on a monitor screen immediately after each measurement in all attempted cases (n = 13). Any residual high pressure zone of the LES was easily recognized and resolved with additional myotomy. Postoperative esophagographies showed resolution of esophageal dilatation. Manometric examination revealed significant reduction of LES pressure in the short-/long term follow-up periods. PH monitoring showed no increase in acid reflux. Overall outcomes were satisfactory (symptom relief = 95%). CONCLUSION: Intraoperative manometry with real-time pressure feedback is a feasible, safe, and useful adjunct in LHD. PMID- 19816022 TI - Pentasomy 49,XXXXY diagnosed in utero: case report and systematic review of antenatal findings. AB - OBJECTIVES: Pentasomy 49,XXXXY is a rare sex chromosome polysomy usually diagnosed postnatally by the combina- tion of mental retardation, facial dysmorphism, and genital, cardiac and skeletal malformations. Prenatal detection of 49,XXXXY is unusual and may be incidental due to non-specific ultrasound (US) findings. We report a case of 49,XXXXY diagnosed prenatally and present a literature review of the few prenatally diagnosed cases. METHODS: We searched the PubMed electronic database without year and language restriction, using the keywords 'Prenatal', 'Diagnosis', and '49,XXXY', performing a systematic review. RESULTS: We report a 35-year-old patient with normal first-trimester US but increased combined risk for trisomies 18 and 13. Amniocentesis at 16 weeks of gestation revealed a 49,XXXXY karyotype. Pregnancy was terminated at 19 weeks' gestation, and a male fetus with facial dysmorphism and hypospadia was delivered. A total of 12 articles were identified in the systematic review. All were case reports and dated from 1980 until 2008. The mean maternal age was 34.8 years (range 30-41). The most common prenatal US feature was cystic hygroma, present in 5 cases. Hypogenitalism was the most common macroscopic clinical feature identified after pathology examination in 7 cases. In 2 cases, there was an increase in first-trimester combined risk for trisomy 21. CONCLUSIONS: Pentasomy 49,XXXXY is associated with a variety of non-specific US findings, of which cystic hygroma was the commonest. No specific sequence of findings could be identified in this review. PMID- 19816023 TI - Amniotic fluid index accuracy in mid-trimester. AB - OBJECTIVE: The aim of this trial was to evaluate the correlation between amniotic fluid index (AFI) and the real amniotic volume in mid-trimester pregnancies. METHODS: Eight women with mid-trimester anhydramnios pregnancies were included. Patients gave their informed consent. Those with premature rupture of membranes were excluded. Amnioinfusion was performed by instilling up to 400 ml of saline solution in 100 ml aliquots. With the patient in supine position, the AFI was determined by the sum of measurements of the deepest vertical pools in each of the 4 quadrants of the maternal uterus, evaluating the AFI after every 100 ml of infused solution. Statistical analysis was performed by analysis of variance, determination coefficient, linear correlation and t test. RESULTS: We realized 29 AFI measurements after amnioinfusions of 100, 200, 300 and 400 ml of saline solution. The AFI ranges that corresponded to the infused volume were: after 100 ml = 5-11 cm (median 6.5 cm); after 200 ml = 9-12.7 cm (median 10.3 cm); after 300 ml = 10-17 cm (median 13.5 cm), and after 400 ml = 12-16 cm (median 15 cm). The correlation between the AFI and the volume infused was r = 0.81 (p < 0.0001), the variance was r(2) = 0.65, suggesting that 65% of the variation in AFI measurements is directly accounted for by the amniotic fluid volume and 35% of the AFI measurements is accounted for by factors other than amniotic volume. CONCLUSIONS: We obtained a strong correlation between AFI and amnioinfusion volume (median of 6.5, 10.3, 13.5 and 15 cm after amniotic infusion of 100, 200, 300 and 400 ml of saline solution, respectively). The variance (0.65) suggests that 35% of AFI measurements is accounted for by factors other than amniotic fluid volume. PMID- 19816024 TI - Total blood volume is maintained in nonhydropic fetuses with severe hemolytic anemia. AB - OBJECTIVE: Fetal alloimmune anemia is associated with increased blood flow velocities and cardiomegaly. In severe cases, hydrops can develop. We investigated whether the decrease of red blood cell volume is associated with a reduction or expansion of plasma volume. METHODS: In 86 alloimmunized fetuses that received a first intrauterine transfusion, we calculated fetal total blood volumes (i.e. fetoplacental blood volumes) using a dilutional principle of fetal hemoglobin with adult hemoglobin. The relation between total blood volume and estimated fetal weight, severity of anemia and hydrops was analyzed. RESULTS: Gestational age ranged from 17 to 35 weeks. Mean hemoglobin deficit was 6.8 standard deviations (range 2.1-11.7) below the normal mean. Fetal total blood volume was significantly related to estimated fetal weight (p < 0.001). Mean total blood volume in nonhydropic fetuses was 123 ml/kg (n = 74) and in hydropic fetuses 144 ml/kg (n = 12). There was a significant relation between total blood volume per kg body weight and hydrops (p = 0.035); however, there was no relation with severity of anemia (p = 0.94). CONCLUSION: In the human nonhydropic fetus with severe hemolytic anemia, total blood volume is maintained: the decrease in red blood cell volume is thus compensated by an increase in plasma volume. In hydropic fetuses, however, total blood volume seems to be increased. This is in accordance with the hypothesis that congestive heart failure plays a role in the pathophysiology of hydrops in anemic fetuses. PMID- 19816025 TI - Development of the fetal duodenum: a postmortem study. AB - OBJECTIVE: To gather detailed morphometric data to be able to assess fetal development of the duodenum and its morphology. MATERIAL AND METHOD: The present study was carried out on 222 human fetuses (114 males, 108 females) aged 9-40 weeks of gestation with no external pathology or anomaly. Fetuses were grouped based on gestational weeks, months and trimesters. Once general external measurements of the fetuses were carried out, the abdominal wall was dissected. The relationships between the duodenum and surrounding structures and its localization were determined. Morphometric data on height, width, length and diameters were gathered. FINDINGS: Means and standard deviations of all parameters were calculated for each gestational week, month and trimester. There were significant relations between measured parameters and gestational age (p < 0.001). There were no sex differences in parameters (p > 0.05). All data were compared with previous studies and discussed. CONCLUSION: Data obtained in this study will contribute to other studies carried out in obstetrics, perinatology, forensic medicine and fetal pathology departments, aimed at identifying anomalies, pathologies and variations of the duodenum and treatment of such cases. PMID- 19816026 TI - Total and fetal cell-free DNA analysis in maternal blood as markers of placental insufficiency in intrauterine growth restriction. AB - OBJECTIVE: To compare total and fetal DNA levels in the maternal plasma in three groups: pregnancies with intrauterine growth restriction (IUGR) due to placental insufficiency (PI) and other causes, and in control pregnancies. METHODS: Total as well as fetal DNA was quantified in 78 maternal plasma samples. In 19 pregnancies, the fetus presented IUGR due to PI (group A), and in 31 pregnancies due to other causes (group B). The control group comprised 28 patients (group C). DNA quantification was done using real-time quantitative PCR with a standardized pool of plasmid calibrators. DNA concentrations of the three groups were compared using non-parametric tests (Kruskal-Wallis or Mann-Whitney tests). RESULTS: The three groups did not statistically differ regarding maternal age (mean +/- SD: 30.5 +/- 5.4 years), gestational age (30 +/- 5.3 weeks) or the proportion of male fetuses (48.2%). Plasma total DNA was significantly higher in group A compared to groups B and C (p = 0.001 for both). An increase in fetal DNA was only observed in group A for patients beyond 28 weeks of gestation. CONCLUSIONS: The plasma total DNA level is higher in patients with IUGR due to PI. These results suggest the presence of maternal endothelial damage independently of preeclampsia. PMID- 19816027 TI - Impact of first-trimester aneuploidy screening in a high-risk population. AB - OBJECTIVE: To determine the impact on maternal decision-making of offering first trimester screening to women at increased risk for fetal aneuploidy. METHODS: A retrospective chart review of all patients > or = 35 years of age at delivery who registered to deliver at Massachusetts General Hospital before (2000-2002; n = 1,604) and after (2003-2005; n = 1,915) the introduction of first-trimester screening. Demographics, the choice of screening test and/or invasive procedure (chorionic villus sampling or amniocentesis), and the detection rate of fetal aneuploidy were compared between the two groups by chi(2) test. Aneuploid newborns without an antenatal diagnosis were reviewed in detail. RESULTS: The fraction of women who accepted prenatal screening and/or an invasive procedure increased from 1,122/1,604 (70%) to 1,751/1,915 (91%, p < 0.01) after the introduction of first-trimester screening. Fewer invasive procedures were done in the second time period, due to a decrease from 40 to 14% in the fraction of women going straight to an invasive procedure without screening (p < 0.01). Similar numbers of aneuploid fetuses were detected prenatally in the two time periods: 21/22 (95%) in the first time period and 22/25 (88%) in the second (p = 0.36). All 4 cases not prenatally diagnosed involved the patient's decision to decline screening or ignore positive screening results. CONCLUSION: The introduction of first-trimester screening in a high-risk population was associated with an increase in the acceptance of prenatal screening and a reduction in the invasive procedure rate without a concomitant decrease in the detection of fetal aneuploidy. PMID- 19816028 TI - Loeys-Dietz syndrome in pregnancy: a case description and report of a novel mutation. AB - Loeys-Dietz syndrome is a syndrome caused by heterozygous mutations in the genes encoding type 1 or 2 transforming growth factor-beta receptor (TGF-beta-R1/2). The obstetrical manifestations are risk of rupture of the gravid uterus and the arteries, either during pregnancy or in the immediate postpartum period, and damage to the vagina, perineum and the colon. We describe, for the first time, a new TGF-beta-R2 gene mutation in a family with several individuals who either had aortic rupture and dissection, sudden death or aortic root dilatation. The pregnancy was followed up and the baby was successfully delivered by a cesarean section at 34 weeks of gestation. The mother's recovery was uneventful and the baby was negative for the mutation on postnatal molecular testing. With appropriate supervision and early delivery by cesarean section, women with Loeys Dietz syndrome can tolerate pregnancy and delivery without any adverse effect. PMID- 19816029 TI - Meconium periorchitis: a rare cause of fetal scrotal cyst--MRI and pathologic appearance. AB - A case of meconium periorchitis detected by fetal MRI and misdiagnosed during pregnancy as inguinoscrotal hernia is reported for the first time. A full-term black boy presented at birth with an asymptomatic, 'stony-hard', scrotal mass suggestive of an in utero testicular torsion or testicular/paratesticular tumor. Early surgical treatment resulted in the removal of paratesticular yellowish amorphous material. Histology was consistent with the diagnosis of meconium periorchitis, a rare and benign condition resulting from healed intrauterine bowel perforation. PMID- 19816030 TI - First trimester diagnosis of sirenomelia by 2D and 3D ultrasound. AB - A case of sirenomelia in a 27-year-old woman detected at 11 weeks 5 days of gestation is presented. It was suspected by two-dimensional sonography and color Doppler imaging. Three-dimensional ultrasound confirmed the final diagnosis. With detailed evaluation of the acquired volumes, valuable information was obtained about the anatomical as well as pathological features of the fetus. PMID- 19816031 TI - Dichorionic triamniotic triplet pregnancy complicated by acardius acormus. AB - This report describes an acardiac fetus of the acormus phenotype in a triplet pregnancy. The diagnosis was confirmed at 15 weeks. In the absence of signs of heart failure in the co-fetus the pregnancy was managed conservatively. The pregnancy was complicated by preterm labour and the fetuses were delivered at 26+5 weeks. The prenatal diagnosis of the acormus phenotype with a well-developed cephalic pole is extremely rare and has never been described antenatally in a higher order multiple pregnancy. We suggest that this rare acardiac fetus phenotype may have a different pathophysiology than those of other phenotypes. The report also summarizes the perinatal outcomes of triplet pregnancies complicated by an acardiac fetus, where the median gestational age at delivery is 26-27 weeks, and discusses the possible therapeutic interventions. PMID- 19816032 TI - Cornelia de lange syndrome: a recognizable fetal phenotype. AB - We describe a fetus with Cornelia de Lange syndrome diagnosed after termination of pregnancy at 21 weeks. Prenatally, growth retardation, diaphragmatic hernia, cystic hygroma and a right hand with only three rays were diagnosed by ultrasound in the second trimester of pregnancy. Postnatal magnetic resonance imaging confirmed the prenatal findings, and the presence of the typical dysmorphic features led to the diagnosis of Cornelia de Lange syndrome. The diagnosis was confirmed by the finding of a truncating mutation in the NIPBL gene. This case illustrates that the diagnosis Cornelia the Lange syndrome can be suspected prenatally in the second trimester, and can be diagnosed in fetuses after induction or newborns at birth as the typical phenotype is present early. PMID- 19816033 TI - Increased fetal nuchal translucency as a sole clue in the prenatal diagnosis of a fetus with trisomy 13. AB - Trisomy 13 is a fatal chromosomal abnormality. A body of literature supports the assessment of fetal nuchal translucency as an adjunct in the prenatal diagnosis of trisomy 13. Over 90% of affected fetuses are readily diagnosed by ultrasound, as they present typical sonographic features; however, there is no consensus regarding trisomy 13 showing normal second trimester sonographic findings. We report an interesting case of trisomy 13, demonstrating increased nuchal translucency as an apparently isolated prenatal sonographic finding. PMID- 19816034 TI - Fetal cardiac troponin T as a marker of poor prognosis in nonimmune hydrops fetalis. AB - OBJECTIVE: Severe fetal anemia and cardiac compromise are important causes of nonimmune hydrops fetalis, and fetal recovery also depends on the degree of fetal heart compromise. The aim of this study was to report the fetal cardiac troponin T (cTnT) levels in cases of nonimmune fetal hydrops. METHODS: Fetal cTnT was analyzed on 7 occasions in 5 cases of nonimmune fetal hydrops (twice in 2 cases). RESULTS: In 3 of 4 fetuses in which intrauterine death occurred, fetal cTnT levels were increased. The only fetus that survived in this series showed decreased levels of cTnT before birth. CONCLUSION: Fetal cTnT levels may be a marker of fetal prognosis in cases of fetal hydrops. PMID- 19816035 TI - Congenital high airway obstruction syndrome due to complete tracheal agenesis: an accident of nature with clues for tracheal development and lessons in management. AB - Congenital high airway obstruction syndrome (CHAOS) is a life-threatening condition characterized by complete blockage of the fetal airways associated with hydrops. We present a case of CHAOS due to the rare cause of complete tracheal agenesis. The ex utero intrapartum therapy (EXIT) strategy was employed to allow for neck and mediastinal exploration. Thymectomy allowed dissection to the level of the carina demonstrating the confluence of dilated mainstem bronchi but no trachea and no connection to the esophagus. A 2.5 endotracheal tube was inserted into the right mainstem bronchus and secured to the left clavicle. At 3 months of age, the infant succumbed to sepsis from Enterobacter mediastinitis due to friction between the tracheostomy tube and the nasogastric tube resulting in erosion of the esophagus. Complete tracheal agenesis, as seen in this case, is consistent with the failure of normal tracheal elongation as suggested by newer theories of foregut development. This case illustrates the most severe form of tracheal atresia causing CHAOS ever salvaged by the EXIT procedure at birth. The subsequent postnatal course highlights the need for early tracheal replacement in this particularly challenging form of CHAOS. PMID- 19816036 TI - Congenital gastric outlet obstruction by pyloric membrane: prenatal and postnatal diagnosis and management. AB - Congenital gastric outlet obstruction is a rare condition representing only 1% of all gastrointestinal atresias. Prenatal diagnosis is uncommon and mostly confined to the third trimester of cases presenting a combination of polyhydramnios with dilated stomach. We report a case of congenital gastric outlet obstruction by pyloric membrane which was diagnosed prenatally in the third trimester by sonography and magnetic resonance imaging. The anomaly appeared to be isolated, thus a favorable outcome was expected. A baby girl weighing 3,430 g was delivered spontaneously at 36 weeks. Postnatal imaging methods confirmed the presence of a congenital gastric obstruction. 21 h after delivery, the baby underwent laparotomy, at which time a malrotation and pyloric membrane were found and resolved. The postoperative course was uneventful and the baby was discharged at the age of 18 days and remains well at controls. PMID- 19816037 TI - Sodium load combined with low doses of exogenous angiotensin II upregulate intrarenal angiotensin II. AB - BACKGROUND/AIMS: The present study was designed to evaluate the effects of a salt load combined with exogenous low nonhypertensive angiotensin II (Ang II) doses on Ang II intrarenal regulation. METHODS: Sprague-Dawley rats were infused with Ang II nonhypertensive doses (0.1 microg.kg(-1).h(-1) and 5 microg.kg(-1).h(-1)) and saline overload (Na 0.5 M, Na 1.0 M and Na 1.5 M) for 2 h (0.04 ml.min(-1)). Sodium tubular reabsorption, sodium urinary excretion and mean arterial pressure (MAP) were measured. Ang II was evaluated in the kidneys by immunohistochemistry. RESULTS: Ang II levels in glomeruli and vessels were exacerbated when sodium load and Ang II were given simultaneously, independently of MAP elevation. In tubules, Ang II staining in the presence of sodium overload was greater in the Ang 0.1 groups than in the Ang 5 groups. Compared with the controls, sodium tubular reabsorption rose in the Ang 0.1-Na 0.5 and Ang 0.1-Na 1 groups and sodium urinary excretion decreased in the Ang 5-Na 0.5 and Ang 5-Na 1 groups. MAP increased in the Ang 5-Na 1 and Ang 5-Na 1.5 groups. CONCLUSION: We conclude that local renal Ang II levels were upregulated when acute sodium overload and nonhypertensive Ang II doses were administered simultaneously in normal rats, independently from blood pressure and glomerular function changes. PMID- 19816038 TI - Gonadectomy influences blood pressure through the kallikrein-kinin system. AB - The kallikrein-kinin system (KKS) appears to be involved in blood pressure regulation. We showed that ovariectomy (oVx) stimulates urinary kallikrein activity (UKa). So, we test whether gonadectomy (Gx) would affect blood pressure through an increase in KKS activity and which mechanism(s) were involved. We studied adult Wistar rats of either sex, with and without Gx. At baseline all groups were normotensive although the oVx mean arterial pressure (MAP) was lower than female MAP (p < 0.05). KKS blockade by aprotinin increased MAP (p < 0.05) exclusively in the oVx group. The probably mechanism(s) involved in KKS regulation (synthesis, renal content and UKa) were also studied. Previous Gx, kallikrein content (nkat/g kidney weight) and UKa (nkat/g kidney weight/day) were higher in female than in male rats: 12 +/- 1.1 versus 6 +/- 0.7 and 40 +/- 6.8 versus 26 +/- 3.4, respectively. After Gx, kallikrein content increased significantly in both orchiectomized (oRx) and oVx rats, and UKa showed a similar tendency (NS). Kallikrein synthesis did not show gender difference in non-Gx rats, but an increase after oVx was observed. KKS was found to be involved in blood pressure regulation in oVx animals. oVx may trigger the increase in kallikrein synthesis and content and UKa to act upon blood pressure. PMID- 19816039 TI - Report of a Brazilian multicenter study on nephropathic cystinosis. AB - INTRODUCTION: The Brazilian Multicenter Nephropathic Study Group, founded in 1999, is currently composed of 16 pediatric nephrology units, which are coordinated by the Pediatric Nephrology Unit of Instituto da Crianca--HCFMUSP. This Study Group intends to better know our patients, their special characteristics and facilitates the treatment. OBJECTIVE: To present an update on the demographics of the ongoing study participants with interest on renal function status, response to therapy, and extra-renal complications. METHODS: Patient recruitment to the study is based on informed consent and has been supported by the Brazilian Society of Nephrology, by the creation of an electronic homepage and by the participation in medical meetings and publications in medical periodicals. Our study protocol involves the initial and follow-up questionnaire, the measurement of intraleukocyte cystine content, initiation and follow-up therapy with cysteamine, and clinical patient follow-up based on a protocol of subsidiary exams. RESULTS: We identified 102 patients (42 females) with nephropathic cystinosis in Brazil since 1999. Forty-six children are followed at the Instituto da Crianca/SP, 15 at the Hospital Pequeno Principe/PR, 12 at the UNICAMP/SP, 10 at the Unidade de Transplante Renal - HCFMUSP/SP and 3 at the Santa Casa/SP; the remaining patients are followed at the Instituto da Crianca and at their respective doctors' offices in different nephrology services in Brazil. Of these patients, 23/102 (22.5%) have normal renal function, 19/102 (18.6%) are in chronic renal failure with conservative treatment, 26/102 are on dialysis (18 on peritoneal dialysis and 8 on hemodialysis), and 34/102 received a renal transplant. The extra-renal involvement diagnosed was: hypothyroidism in 63 patients, diabetes mellitus in 8 patients, muscular involvement in 7 patients, a compromised central nervous system in 5 patients, hepatic complications in 5 patients, and deglutition dysfunction in 2 patients. During this period, 10/102 patients died. Cysteamine has been used by 81/102 patients (20 children started the therapy under 2 years of age). Growth parameters were improved by cysteamine, mainly in the youngest patients. We used recombinant growth hormone in 15 patients with persistent low growth velocity and stature z score under 2.5%. We could also observe a delay in appearance of extra-renal complications in patients receiving cysteamine. CONCLUSION: Our study demonstrates the importance of a multi-center study for recruitment, diagnosis and management of rare diseases. This study promotes access to the adequate treatment with profound impact on the quality of life. PMID- 19816040 TI - Inpatient hemodialysis initiation: reasons, risk factors and outcomes. AB - BACKGROUND/AIMS: Inpatient initiation of chronic hemodialysis is considered undesirable because of cost and possible harms of hospitalization. We examined the patient characteristics and outcomes associated with inpatient initiation. METHODS: In a prospective cohort study of incident dialysis patients, the independent association of inpatient hemodialysis initiation with patient outcomes was assessed in multivariable analyses with adjustment for patient characteristics and propensity for inpatient initiation. RESULTS: A total of 410 of 652 (63%) hemodialysis patients began as inpatients; uremia and volume overload were the most commonly documented reasons. Compared to outpatients, inpatients were more likely to be unmarried, report less social support, have multiple comorbidities and be referred to a nephrologist 4 months or less prior to initiation. Inpatient initiation was protective for subsequent all-cause hospitalization (incidence rate ratio (IRR) = 0.92, confidence interval (CI) 0.89 0.94); this was most pronounced among those who had the highest propensity for inpatient initiation (IRR = 0.66, CI 0.56-0.78), including those referred late to nephrology. Similar results were found for infectious hospitalization. Mortality [hazard ratio = 1.03, CI 0.82-1.30] and cardiovascular events were not significantly different for inpatients versus outpatients. CONCLUSION: Inpatient hemodialysis initiation has a protective association with hospitalization among those patients referred late to nephrology, with multiple comorbidities and/or little social support. PMID- 19816041 TI - A small reduction in the ankle-brachial index is associated with increased mortality in patients on chronic hemodialysis. AB - BACKGROUND: High pulse wave velocity (PWV) and a low ankle-brachial index (ABI) are associated with mortality in hemodialysis (HD) patients. Recently, the cardio ankle vascular index (CAVI) was developed as a novel index of arterial stiffness independent of blood pressure. METHODS: We compared brachial-ankle PWV (baPWV), the ABI and the CAVI as predictors of mortality in 194 HD patients (age 64 +/- 12 years; time on HD 111 +/- 96 months) during a follow-up period of 39 +/- 4 months (range 31-46). RESULTS: The ABI was significantly positively correlated with serum albumin and negatively with log-transformed highly sensitive C-reactive protein (p < 0.01), while baPWV and the CAVI were not. Of 194 patients, 39 patients (20.1%) died during the follow-up, 25 (64.1%) of cardiovascular causes. Kaplan-Meier analysis revealed that the patients with an ABI in the lowest tertile (<1.0) had a significantly lower survival rate (p < 0.01). Cox hazards analysis after adjustment for the conventional risk factors revealed that an ABI value in the lowest tertile was a determinant of total mortality when compared with ABI values in the highest tertile [>1.1; hazard ratio 3.50 (95% confidence interval 1.20-10.20); p = 0.02]. In contrast, baPWV and the CAVI were not associated with mortality. CONCLUSION: These findings suggest that a small reduction in the ABI (<1.0) is an independent predictor of all-cause mortality in chronic HD patients. PMID- 19816042 TI - Vitamin B6 metabolism in chronic kidney disease--relation to transsulfuration, advanced glycation and cardiovascular disease. AB - BACKGROUND: Vitamin deficiency is common in chronic kidney disease (CKD). Data on B(6) supply and possible relationships to cardiovascular events (CVE) in CKD are rare. Pyridoxamine exerts inhibitory effects on the formation of advanced glycation endproducts (AGE) implicated in the pathogenesis of CKD and atherosclerosis. METHODS: In 48 CKD patients at stage 2-4, 72 hemodialysis patients (HD), 38 renal transplant recipients (RTR) and 141 healthy controls (mean age 58 +/- 13, 61 +/- 12, 50 +/- 12 and 54 +/- 16 years, respectively), plasma and red blood cell (RBC) concentrations of pyridoxal-5'-phosphate (PLP), pyridoxal (PL), 4-pyridoxic acid (PA), pyridoxamine-5'-phosphate (PMP) and of the AGE pentosidine were measured by high-performance liquid chromatography, N(epsilon)-(carboxymethyl)lysine and imidazolone by an ELISA, and total homocysteine and cystathionine by gas chromatography-mass spectrometry. RESULTS: Despite routine low-dose vitamin supplementation in HD, plasma PLP was decreased in HD (79 +/- 69 nmol/l) compared with CKD stage 2-4 patients (497 +/- 944 nmol/l), RTR (416 +/- 604 nmol/l) and controls (159 +/- 230 nmol/l; p < 0.001). Plasma PA was significantly increased in HD (11,667 +/- 17,871 nmol/l) in comparison with CKD stage 2-4 (435 +/- 441 nmol/l), RTR (583 +/- 668 nmol/l) and controls (46 +/- 49 nmol/l; p < 0.001). B(6) forms were significantly affected by renal function (R = 0.792, p < 0.001 for CKD stage 2-4). There was no relation of vitamers with a history of CVE. Relationships between B(6) forms and AGE (RBC-PMP with pentosidine in CKD stage 2-4: R = -0.351, p < 0.05) were found. CONCLUSION: HD patients showed a deficiency in PLP in plasma but not in RBC. Prospective trials are needed to elucidate the potential role of elevated PA on cardiovascular and renal outcome in CKD. Vitamin B(6) supplementation might be successful in preventing AGE-related pathologies. PMID- 19816044 TI - Does diabetes mellitus predispose to increased fluid overload in peritoneal dialysis patients? AB - BACKGROUND: Diabetic peritoneal dialysis patients have been reported to have faster peritoneal solute transport and may be at risk of reduced ultrafiltration volumes, leading to fluid overload. METHODS: We audited multi-frequency bioimpedance data from 198 consecutive peritoneal dialysis patients (141 nondiabetics and 57 diabetics). RESULTS: Diabetic patients had increased body mass index (males 27 +/- 4 vs. 26 +/- 4; females 28 +/- 5 vs. 25 +/- 4; p < 0.01 for both), waist to hip ratio (males 0.94 +/- 0.07 vs. 0.94 +/- 0.04; females 0.99 +/- 0.9 vs. 0.94 +/- 0.09; p < 0.01 for both) and body fat (males 31 +/- 8 vs. 26 +/- 11%, p < 0.05; females 40 +/- 8 vs. 33 +/- 10%, p < 0.01) compared to nondiabetic patients. D4/P creatinine was greater for the female diabetic patients than the nondiabetic patients (0.75 +/- 0.1 vs. 0.69 +/- 0.1; p = 0.044); otherwise, the peritoneal equilibrium test (PET) results did not differ. Extracellular water (ECW) adjusted for height was similar in diabetic and nondiabetic patients, but the ratio of ECW to total body water (TBW) was greater for diabetics (males 0.40 +/- 0.01 vs. 0.39 +/- 0.01; females 0.39 +/- 0.01 vs. 0.38 +/- 0.01; p < 0.05 for both). CONCLUSIONS: Diabetic and nondiabetic patients had similar PET results, 24-hour net peritoneal ultrafiltration and blood pressure control. The ratio of ECW to TBW was greater in diabetic patients, and although this could be due to a loss of intracellular water, as albumin and C reactive protein did not differ, it suggests that diabetic peritoneal dialysis patients have an expanded extracellular volume. PMID- 19816043 TI - Improving the management of chronic kidney disease in Uruguay: a National Renal Healthcare Program. AB - BACKGROUND: Uruguay has implemented a chronic kidney disease (CKD) prevention program. AIMS: The objectives of the study are to assess the results of the National Renal Healthcare Program (NRHP). METHODS: This study is a cohort study of nondialysis-registered patients from October 2004 to March 2008. We made a comparison between patients under nephrology care (NC) or the care of a primary care physician (PCP; prereferral). In the outcome analysis, the primary endpoint was end-stage renal disease (ESRD) and the secondary endpoints were progression of CKD, compliance with the therapeutic goals and death. ESRD/mortality predictors were determined by Cox analysis. RESULTS: The study comprised 2,219 patients aged 67.4 +/- 13.5 years, of whom 52.5% were male, 42.1% hypertensive, 16.9% had diabetic nephropathy, and 61.3 and 21.4% were in CKD stages III and IV, respectively. At baseline, NC patients showed a better control than patients under the care of a PCP: systolic blood pressure > or =160 mm Hg (22.4 vs. 31.1%); total cholesterol <5.8 mmol/l (56.6 vs. 42.5%); and low-density lipoprotein cholesterol <2.9 mmol/l (41.2 vs. 29.1%). Control improved in patients under the care of a PCP according to years of enrollment. Outcome analysis (1,188 patients) showed a significant improvement in targets, with 56% of the patients stabilizing. CKD stage IV, diabetic nephropathy, proteinuria and hypertension increased the risk of ESRD; angiotensin-converting enzyme inhibitors/angiotensin receptor blockers and age <65 years decreased the risk. CONCLUSIONS: Our results highlight the best management of CKD patients in both groups and the impact of the NC and renin-angiotensin-aldosterone system blockers. PMID- 19816046 TI - Role of cyclooxygenase and derived reactive oxygen species in rho-kinase-mediated impairment of endothelium-dependent vasodilation and blood flow after ischemia reperfusion of the rat kidney. AB - BACKGROUND/AIMS: Decreased endothelium-dependent vasodilation and blood flow in renal ischemia-reperfusion (IR) may result in part from rho-kinase activation, and cyclooxygenase (COX) activation, and resultant reactive oxygen species (ROS) may be involved. METHODS: We tested this hypothesis in male Wistar rats, subjected to 60 min of bilateral clamping of the renal arteries and 60 min of reperfusion or a sham procedure, and treated by the rho-kinase inhibitor Y27632 (1 mg/kg) and/or the nonspecific COX inhibitor diclofenac (10 mg/kg). Renal blood flow was measured by fluorescent microspheres, and ROS in the arterial endothelium was quantified by dihydroethidium staining. Endothelium-dependent vasodilation was determined by an acetylcholine concentration-response curve in the presence or absence of diclofenac (10 microM). RESULTS: Y27632 increased renal blood flow and reduced ROS in vivo, and improved endothelium-dependent vasodilation in vitro, following IR with or without diclofenac. Following IR, diclofenac had no effect on renal blood flow and ROS in vivo, but improved endothelium-dependent vasodilation in vitro. CONCLUSION: Activation of rho-kinase impairs endothelium-dependent vasodilation and perfusion following renal IR, independently of COX and resultant ROS. In contrast, the vasodilatory effect of rho-kinase inhibition may be partly mediated by decreasing ROS, unrelated to COX and resultant vasoconstricting prostanoids. PMID- 19816045 TI - Evaluation of oxidant-antioxidant balance in patients on maintenance haemodialysis: a comparative study of dialyzers membranes. AB - BACKGROUND: Haemodialysis (HD) exacerbates oxidative stress (OS). The polymethyl methacrylate (PMMA)-BK-F membrane ameliorates OS and inflammation markers compared to polyacrylonitrile (PAN/AN69) and cellulose membranes. This may be due to the size of pore radius, high flux or other specific properties of PMMA membranes. AIM: To compare OS and inflammatory status in HD-treated end stage renal disease patients with membranes of different pore size radius and flux. METHODS: 47 patients of both sexes were studied. The HD membranes with which the patients were normally treated were changed to BK-P or B-3 membranes for 6 months. Intracellular and extracellular components of the oxidant-antioxidant balance (OAB), C-reactive protein (CRP), beta2-micro-globulin (beta2mu-globulin), albumin and transferrin were measured. RESULTS: A significant decrease in red cell membrane thiobarbituric acid reacting substances and an increase in cytosolic superoxide dismutase (SOD) and plasma total antioxidant substances were observed in all patients after 6 months of treatment with BK-P and B-3 membranes except SOD and CRP in patients previously dialysed with triacetate cellulose membranes. Albumin and transferrin remained unmodified. beta2mu-globulin significantly decreased after treatment with PMMA membranes. CONCLUSION: BK-P and B-3 HD membranes improved the OAB, beta2mu-globulin and CRP compared to PAN/AN69 and cellulose diacetate membranes. PMID- 19816047 TI - Multi-glycoside of Tripterygium wilfordii Hook f. ameliorates prolonged mesangial lesions in experimental progressive glomerulonephritis. AB - BACKGROUND/AIMS: Multi-glycoside from Tripterygium wilfordii Hook f. (GTW) is used for treatment of progressive glomerulonephritis (GN) in China. We have previously reported the beneficial effects of GTW on acute GN induced by an anti Thy-1.1 monoclonal antibody (mAb). In the present study, the effect and potential mechanisms of GTW on the chronic irreversible model of GN were investigated. METHODS: Progressive GN was induced in rats by two intravenous injections of anti Thy-1.1 mAb 1-22-3. Daily oral administration of GTW was started before the second injection of mAb until the day of sacrifice. Ten rats were randomly divided into a control (vehicle-treated) and a GTW-treated group, and sacrificed on day 45 after the first injection of mAb 1-22-3. Proteinuria was determined on days 0, 1, 3, 5, 7, 10, 14, 20, 25, 30, 35, 40, and 45. Blood biochemical parameters, morphological changes of mesangium, glomerular infiltration of macrophage and T lymphocyte, and glomerular mRNA expression of cytokines (TGF beta, IL-2, and IFN-gamma) were examined from the samples taken at terminal sacrifice. RESULTS: GTW treatment significantly ameliorated proteinuria, renal function, prolonged mesangial lesions and inflammatory cell accumulation in glomerulus. In addition, it significantly reduced the glomerular mRNA expression for TGF-beta, IL-2, and IFN-gamma. CONCLUSION: GTW ameliorates prolonged glomerular lesions presumably through suppression of cytokine production (TGF beta, IL-2, and IFN-gamma). GTW could be an effective therapeutic agent for treatment of chronic renal diseases. PMID- 19816048 TI - Sp1 specifically binds to an evolutionarily conserved DNA segment within a region necessary for podocyte-specific expression of nephrin. AB - We have analyzed a conserved 237-bp segment located in a 1.9-kb upstream region of the nephrin gene, previously shown to contain kidney specific enhancer element(s). Electromobility shift assay was used to identify a 20-nucleotide region specifically recognized and bound by protein factors in nuclear extracts from immortalized podocyte and human embryonic kidney cell lines. The region was further narrowed down by competition assays to a stretch of 6 consecutive guanines, which are conserved at this location in multiple species. Introduction of mutations in this sequence abolished all protein binding activity whereas mutations in the flanking nucleotides did not. By means of gel supershift and chromatin immunoprecipitation assays we have shown that the protein factor from podocyte nuclear extracts able to recognize and bind the target sequence is the Sp1 zinc-finger protein. PMID- 19816049 TI - BMP-7 blocks the cyclosporine-A-induced epithelial-to-mesenchymal transition in renal tubular epithelial cells. AB - AIMS: The nephrotoxic side effects of cyclosporine A (CsA) are partly due to induction of the epithelial-to-mesenchymal transition (EMT), possibly through upregulation of connective tissue growth factor (CTGF). Bone morphogenetic protein (BMP-7) has been reported to protect against and reverse renal injury via its renotropic and antifibrotic effects. We therefore designed a method to investigate the mechanism by which BMP-7 inhibits CSA-induced EMT in cultured human renal proximal tubular epithelial (HK-2) cells and whether BMP-7 can antagonize CsA-induced profibrogenic effects. METHODS: Cultured HK-2 cells were treated with CsA or a combination of CsA and BMP-7 for 72 h. Morphological changes were assessed by phase-contrast microscopy. The expression of alpha smooth muscle actin (alpha-SMA), E-cadherin, collagen type I and CTGF was analyzed by immunofluorescence, RT-PCR, and Western blot. Additionally, the effect of CTGF silencing on CsA-mediated gene expression was assessed. RESULTS: CsA-induced EMT was associated with decreased expression of E-cadherin, increased expression of alpha-SMA, collagen type I and CTGF, and loss of epithelial morphology. BMP-7 inhibited these effects in a dose-dependent manner. Using siRNA, CTGF knock-down also attenuated EMT-associated phenotypic changes induced by CsA. CONCLUSION: These data suggest that BMP-7 can block CsA-induced EMT by downregulating the expression of CTGF, a downstream mediator of EMT. PMID- 19816050 TI - Vasopressin type 2 receptor V88M mutation: molecular basis of partial and complete nephrogenic diabetes insipidus. AB - BACKGROUND/AIMS: Mutations in the type 2 vasopressin receptor gene (AVPR2) underlie X-linked recessive nephrogenic diabetes insipidus (NDI). Here, we report on a family with a mutation in AVPR2, c.262G>A (p.V88M). This recurrently identified mutation was previously shown to abolish AVPR2 function, yet in some affected members, urine osmolalities of up to 570 mosm/kg were observed. We detail the variable clinical phenotype and investigate its molecular basis. METHODS: Retrospective analysis of clinical data and in vitro assessment of wild type and V88M-mutant receptors. RESULTS: Clinical data were available on 6 patients. Four of these demonstrated a substantial increase in urinary concentration after 1-desamino[8-D-arginine] vasopressin, consistent with partial NDI, while 2 did not respond. In vitro analysis revealed a reduced cell surface expression and decreased binding affinity for arginine-vasopressin of the mutant receptor, leading to blunted signaling activity. Treatment with the pharmacological chaperone SR121463 enhanced cell surface expression. CONCLUSION: The V88M mutation is associated with phenotypical diversity, which may be explained by the fact that both the expression level and the hormone-binding affinity are affected by the mutation. Our results provide a rational basis for treatment trials with vasopressin analogues in combination with pharmacologic chaperones in patients with this recurrently identified mutation. PMID- 19816051 TI - The impact of high anxiety level on cellular and humoral immunity in mice. AB - BACKGROUND/AIMS: In the present study, we aimed to examine whether a high anxiety level affects various parameters of immunity in mice. METHODS: We used the behavioral light/dark choice test to evaluate whether high anxiety has an impact on various parameters of cellular (granulocytes, monocytes, total lymphocytes, TCD4(+), TCD8(+) and NK numbers) and humoral (IgA, E and G concentrations) immunity. Secondly, we investigated whether the cellular and humoral immune systems of mice with contrasting levels of anxiety responded differently to stressors (such as physical restraint) by monitoring blood markers of the both types of immunity. RESULTS: High levels of anxiety inhibited part of the cellular and humoral immune systems by significantly decreasing total lymphocytes numbers (including TCD4(+) and TCD8(+)) and immunoglobulin (A and E) concentrations. However, no significant changes in the number of granulocytes, monocytes or NK cells were observed. As a consequence, overall, our results suggest that high anxiety led to a decrease in the efficiency of the immune system of anxious mice. On the other hand, our findings also showed that restraint stress (acute and subacute) produced the same immunological profile as high anxiety in mice. This was independent of the animals' anxiety status. At the same time, we observed that restraint stress produced significant increases in the levels of granulocytes and monocytes. CONCLUSIONS: High anxiety and restraint stress exerted adverse effects on cellular and humoral immunity in mice. While the effect of restraint stress was independent of the anxiety levels in mice, this stress led to an aggravation of the immune response from the high degree of anxiety. Therefore, anxious subjects could be more vulnerable to infections and inflammation, particularly when they are exposed to stressful situations. PMID- 19816052 TI - Neuroendocrine and immunological correlates of chronic stress in 'strictly healthy' populations. AB - BACKGROUND: Chronic stress has been associated with detrimental or maladaptive neuroendocrine and immunological changes. OBJECTIVES: We assessed the neuroendocrine and immunological correlates of a realistic chronic stress experienced by strictly healthy caregivers of Alzheimer's disease patients and age-matched controls. METHODS: We screened 330 caregivers and 206 non-caregivers according to the 'strictly healthy' conditions established by the SENIEUR protocol. Forty-one strictly healthy caregivers (60.56 +/- 16.56 years) and 33 non-stressed controls (60.27 +/- 14.11 years) were selected for this study. Salivary cortisol and dehydroepiandrosterone sulfate (DHEAS) were assessed at multiple points by radioimmunoassay. Peripheral T cell proliferation and cellular sensitivity to glucocorticoids (corticosterone and dexamethasone, DEX) were evaluated by colorimetric assays. We also examined the hypothalamic-pituitary adrenal (HPA) axis response to the administration of a low-dose DEX in vivo. RESULTS: The caregivers were significantly more stressed, anxious and depressed than non-caregivers (all p < 0.0001), in contrast to similar cortisol levels. Caregivers had reduced DHEAS levels (-32%, p < 0.0001), an increased cortisol/DHEAS ratio (39.7%, p < 0.0001) and impaired HPA axis response to DEX intake. Caregivers had a higher T cell proliferation (p < 0.0001) and increased sensitivity to glucocorticoids in vitro (p < 0.01) as compared to non-stressed controls. CONCLUSIONS: Our results suggest that the maintenance of health in chronically stressed populations may be associated with both protective and detrimental neuroendocrine and immunological changes. PMID- 19816053 TI - Levels of IL-1beta and IL-1ra in cerebrospinal fluid of human patients after single and prolonged seizures. AB - BACKGROUND: Experimentally induced seizures are associated with increased production of inflammatory cytokines in the nervous system. Elevated cerebrospinal fluid levels of cytokine interleukin-6 (IL-6) have been found after a single generalized seizure in human patients. After prolonged seizures, levels of IL-6 have been shown to be even higher compared with single seizures. In the present study, we determined the levels of proconvulsive IL-1beta and anticonvulsive IL-1ra in cerebrospinal fluid after single tonic-clonic seizures as well as after prolonged seizures. METHODS: The levels of cytokines were measured using enzyme-linked immunosorbent assay. RESULTS: We found that after single seizures, a slight increase in anticonvulsive IL-1ra levels was found; however, after prolonged partial or recurrent tonic-clonic seizures, the levels of IL-1ra were significantly elevated, together with decreased IL-1beta levels. CONCLUSION: Our results indicate that after severe seizures, the balance between IL-1-type cytokines is changed towards a neuroprotective and anticonvulsive direction with an overproduction of IL-1ra with respect to potentially neurotoxic IL-1beta. This reaction may serve as a defense mechanism of the nervous system against excitotoxic neuronal damage. PMID- 19816054 TI - Effects of methylprednisolone and glatiramer acetate on nitric oxide formation of cytokine-stimulated cells from the rat oligodendroglial cell line OLN-93. AB - OBJECTIVES: In multiple sclerosis, an autoimmune inflammatory disease, oligodendroglia are primarily affected and play an important role in the onset and process of the degeneration of neuronal axons. High-dose therapy with glucocorticoids like 6alpha-methylprednisolone (MP) as well as the application of immunomodulatory agents like glatiramer acetate (GA) are commonly used in the treatment of MS. The purpose of our study was to examine, in an adequate cell culture model, the effect of MP and GA on oligodendroglial activation induced by pro-inflammatory stimuli. METHODS: In the present study, we measured the mRNA (real-time RT-PCR) and protein (Western blot) expression of inducible nitric oxide synthase (iNOS) and the release of nitric oxide (NO; Griess reagent) of rat oligodendroglial progenitor cell line OLN-93 after pro-inflammatory stimulation, and searched for influences of MP and GA on these parameters. OLN-93 cells were treated either with a combination of TNF-alpha and IFN-gamma alone, or additionally with MP or GA. Cell viability and cell protein contents were determined in parallel. RESULTS: Our results show that TNF-alpha and IFN-gamma increased iNOS mRNA and protein expression and the NO production in OLN-93 cells. The elevated production of NO and iNOS protein was reduced in the presence of MP, whereas under treatment with GA, the cytokine-induced overproduction of NO did not change significantly. CONCLUSIONS: The presented data suggest an active role of oligodendroglial cells in inflammatory processes like multiple sclerosis and indicate different properties of MP and GA regarding immunosuppression. PMID- 19816055 TI - Kainic acid-activated microglia mediate increased excitability of rat hippocampal neurons in vitro and in vivo: crucial role of interleukin-1beta. AB - OBJECTIVE: It is known that highly activated microglia and the consequent production of inflammatory cytokines were associated with neuroexcitotoxic injuries. The present study was carried out to explore whether interleukin-1beta (IL-1beta), a proinflammatory cytokine produced in abundance by activated microglia, mediates increased excitability of hippocampal neurons and the related molecular mechanisms. METHODS: Primary cultured microglia were activated by kainic acid (KA), and the KA-treated microglial conditioned medium (KA-MCM) was collected. KA-MCM with or without anti-rat IL-1beta monoclonal neutralizing antibody was then injected into the rat in the right cerebral ventricle, or primary cultured hippocampal neurons were treated with the above-mentioned KA MCM. The population spike amplitude changes in the CA3 region were assessed by electrophysiological recording in vivo. Western blot and RT-PCR assay were performed to investigate the expression changes of N-methyl-D-aspartate receptor subunit 1 (NMDAR1) and inducible nitric oxide synthase (iNOS) expression in hippocampal neurons. RESULTS: Primary cultured microglia were significantly activated by KA with increased IL-1beta levels. Interestingly, intracerebroventricular administration of KA-MCM to rats resulted in enhancement of population spike amplitude in the CA3 region and in upregulation of NMDAR1 and iNOS expression in the hippocampus, which was partially attenuated by anti-rat IL 1beta antibody. Furthermore, the changes in NMDAR1 and iNOS expression in the rat hippocampus were verified by incubation of primary cultured hippocampal neurons with KA-MCM. CONCLUSION: This study provides evidence that KA-activated microglia mediate increased excitability of hippocampal neurons in vitro and in vivo and that IL-1beta may be one of the main causes of this event. PMID- 19816056 TI - Inflammatory biomarkers in patients with posttraumatic stress disorder caused by myocardial infarction and the role of depressive symptoms. AB - OBJECTIVE: Inflammation might link posttraumatic stress disorder (PTSD) with an increased risk of cardiovascular events. We explored the association between PTSD and inflammatory biomarkers related to cardiovascular morbidity and the role of co-morbid depressive symptoms in this relationship. METHODS: We investigated 15 patients with interviewer-rated PTSD caused by myocardial infarction (MI) and 29 post-MI patients with no PTSD. All patients completed the depression subscale of the Hospital Anxiety and Depression Scale and had blood collected to determine inflammatory markers of increased cardiovascular risk. RESULTS: Controlling for demographic and medical covariates, patients with PTSD had higher leptin levels than patients with no PTSD (p = 0.038, explained variance 10.4%); this difference became nonsignificant when controlling for depressive symptoms. After controlling for depressive symptoms, PTSD patients had higher interleukin-6 (p = 0.041; explained variance 10%), lower C-reactive protein (p = 0.022, explained variance 12.1%), and lower soluble CD40 ligand (p = 0.016, explained variance 13.4%) than patients without PTSD. After controlling for PTSD status, depressive symptoms correlated with soluble CD40 ligand (r = 0.45, p = 0.002) and with C-reactive protein (r = 0.29, p < 0.07). CONCLUSIONS: The findings provide further evidence for altered inflammation in PTSD. Comorbid depressive symptoms ought to be considered to disentangle the unique associations of PTSD caused by MI and systemic inflammation. PMID- 19816057 TI - Prenatal lipopolysaccharide exposure affects maternal behavior and male offspring sexual behavior in adulthood. AB - OBJECTIVE: This study investigates the effects of prenatal lipopolysaccharide (LPS) exposure on the maternal behavior of pregnant rats and the physical development and sexual behavior of their male offspring in adulthood. METHODS: For two experiments, pregnant rats were injected with LPS (250 microg/kg, i.p.) on gestation day (GD) 21. In the first experiment, the maternal behavior (postnatal day, PND, 6) and the dam's open-field general activity (PND7) were evaluated. In the second experiment, the maternal pre- and postnatal parameters, the pup's development, the offspring's sexual behavior in adulthood, and the pup's organ weights were assessed. RESULTS: Compared to the control group, the LPS-treated dams presented reduced maternal behavior, decreased general activity, a smaller body weight difference between GD21 and PND1, a greater number of perinatal deaths, and smaller litters. For the male pups, LPS treatment resulted in a decreased body weight on PND2, whereas the anogenital distance and the day of testis descent were not modified. The male sexual behavior was impaired by prenatal LPS. Particularly the number of ejaculating animals was reduced. The testis weight was also lower in the prenatally LPS-treated rats than in the control rats. CONCLUSION: We propose that prenatal LPS exposure on GD21 acts as an imprinting factor that interferes with the programming of brain sexual determination in offspring. PMID- 19816058 TI - Distribution of inducible nitric oxide synthase and tumor necrosis factor-alpha in the peripheral nervous system of Lewis rats during ascending paresis and spontaneous recovery from experimental autoimmune neuritis. AB - BACKGROUND: Inducible nitric oxide synthase (iNOS) and tumor necrosis factor alpha (TNF-alpha) are pleiotropic molecules with widespread action in autoimmune diseases. OBJECTIVE: This study characterizes the distribution of iNOS and TNF alpha in the spinal nerve roots, dorsal root ganglia and sciatic nerve of Lewis rats during experimental autoimmune neuritis (EAN). METHODS: Macrophages and neutrophils were identified by immunofluorescence as cellular sources of iNOS and TNF-alpha at various stages of EAN induced by synthetic peptide 26. RESULTS: As the disease progressed, iNOS- and TNF-alpha-bearing cells gradually infiltrated the cauda equina, dorsal root ganglia, Th12-L3 spinal roots, and the sciatic nerve. A severer EAN profile developed when more iNOS- and TNF-alpha-bearing cells were present, and the recovery from EAN was related to the disappearance of these cells and the regeneration of nerve fibers. CONCLUSIONS: This is the first report to show iNOS- and TNF-alpha-immunoreactive cells in dorsal root ganglia during EAN, suggesting an underlying pathology for the neuropathic pain behavior in EAN. Our results suggest that the cells bearing iNOS and TNF-alpha in the different parts of the peripheral nervous system are involved in the development of the clinical signs observed at each stage of EAN. PMID- 19816059 TI - Centre for neuroscience: an exploration of signalling in neural development, health and disease. PMID- 19816060 TI - Signals involved in neural differentiation of human embryonic stem cells. AB - Neural differentiation from embryonic stem cells involves progressive stages of neural induction, expansion and maintenance of neural stem/progenitor cells, and differentiation to neurons and glia. Our understanding of the signals involved in each of these processes is primarily based on our knowledge of neural development during embryogenesis. This review will focus on the signalling pathways that have been identified to play a role in neural differentiation of human embryonic stem cells (hESCs), including their induction to neuroectoderm, maintenance and expansion of hESC-derived neurospheres, differentiation to neurons and specification to specific neuronal lineages. Understanding the signals involved in each of these stages is important for optimising methods to derive specific cell types for transplantation therapies, as well as for providing insight into the mechanisms of human neurogenesis. PMID- 19816061 TI - Regulation of stem cell pluripotency and neural differentiation by lysophospholipids. AB - Lysophospholipids are bioactive signalling molecules able to act through the binding of their specific G-protein-coupled receptors to exert pleiotropic effects on a wide range of cells. The most widely studied signalling lysophospholipids are lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P). LPA and S1P have been identified to have widespread developmental, physiological and pathological actions in the central nervous system and more recently have been shown to induce biological effects on various stem cell types. This review aims to summarise the current knowledge on LPA and S1P regulation of embryonic and neural stem cell biology. PMID- 19816062 TI - Effects of bone morphogenic proteins on neural precursor cells and regulation during central nervous system injury. AB - Bone morphogenic proteins (BMPs) are well known for their influence on cell fate determination, proliferation and differentiation during early embryogenesis. Here, we review evidence for BMPs playing an additional, ongoing role in the proliferation and differentiation of neural precursor and progenitor cells in postnatal and adult central nervous system (CNS) and in CNS injury. The effects of BMPs on CNS cells have been studied using primary cultures of neural precursor and oligodendrocyte lineage cells. In addition, transgenic mice have been used to investigate in vivo effects of altering BMP pathway activation, and rodent models of CNS injury have been used to examine endogenous regulation of BMPs. These results have shown that BMPs promote production of astrocytes and inhibit production and maturation of oligodendroglia. The effects of BMPs on neurogenesis could be dependent on the origin of precursor cells or on the specifics of the microenvironment of the cell niche, as there are reports of inhibition and promotion of neurogenesis by BMPs. There is emerging evidence that BMPs are upregulated in several models of CNS injury; however, the effects of this regulation have not been well characterised. Understanding of the function of endogenous BMP regulation is important for determining how modulation of BMP signalling could improve repair following CNS injury. PMID- 19816063 TI - The role of neurotrophins in the regulation of myelin development. AB - Neurotrophins comprise a family of growth factors that are expressed in a variety of cell types, and which exert influences on a large range of cellular activities that are important for development and the maintenance of the nervous system, as well as in neurodegenerative and psychiatric disorders. More recently, neurotrophins have been implicated in influencing the dynamic and complex signals that occur between neurons and glial cells, including Schwann cells in the peripheral nervous system and oligodendrocytes in the central nervous system that regulate myelination. Here we review the recent studies that identify neurotrophins as important regulators of both peripheral and central myelination, highlight some of the many questions that remain to be answered, and identify possibilities for further research. PMID- 19816064 TI - TAM receptor signalling and demyelination. AB - The TAM family (Tyro3, Axl and Mer) of receptor protein tyrosine kinases play pivotal roles in a number of major cellular processes: cell survival and proliferation, immunomodulation and phagocytosis. These processes are central to both the initial development and pathological course of human multiple sclerosis. All three receptors and their ligands, Gas6 (growth arrest-specific gene 6) and protein S, are expressed in the central nervous system (CNS), including in oligodendrocytes, the myelin-producing cell of the CNS. Recent studies have shown that Gas6-dependent TAM receptor signalling is an important modulator of oligodendrocyte survival and microglial phenotype both in vitro and in vivo. Multiple lines of evidence allow us to hypothesise that, during a demyelinating challenge, dysfunctional TAM receptor signalling could lead to a 'vicious cycle' of cell death, reduced phagocytosis and deleterious immune hyper-activation. A current challenge in this field is to expand our understanding of TAM receptor signalling from rodent models of central demyelination to human disease. PMID- 19816065 TI - PCTK proteins: the forgotten brain kinases? AB - PCTAIRE kinases (PCTKs) are highly conserved serine/threonine kinases that are closely related to cyclin-dependent kinases. They are enriched in post-mitotic neurons of adult brains, suggesting they might perform important neuron-specific functions independent of the cell cycle. So far, the biological functions of PCTKs in the brain have been largely neglected and remain to be discovered. This review summarises preliminary investigations into the expression and characterisation of PCTK kinase activity, providing a basis for further investigations. In particular, it identifies three key areas of priority for further research: (1) do PCTKs require activating phosphorylation by an upstream kinase or binding to an essential co-factor for maximal kinase activity; (2) what are the physiological substrates of PCTKs, and (3) what is the functional effect of PCTK kinase activity in primary neurons? PMID- 19816066 TI - Muscarinic receptors in psychiatric disorders - can we mimic 'health'? AB - The concept that acetylcholine is involved in the pathophysiologies of psychiatric disorders has existed since the 1950s. There is very strong evidence implicating a dysfunctional muscarinic system in schizophrenia, +with less information available for bipolar disorder and major depressive disorder. The translation of this evidence into clinically viable treatments has been disappointing; hampered by problems associated with developing drugs that target the requisite members of the muscarinic family, rather than all of the receptors, which results in unacceptable side-effect profiles. The discovery of additional binding sites, other than the one occupied by acetylcholine, has revitalised research into this aspect of psychopharmacology. New compounds are now being developed that have the potential to selectively target individual muscarinic receptors in the central nervous system. The question that remains to be answered is whether stimulating central muscarinic receptors will result in the reestablishment of normal central muscarinic activity? The purpose of this review is to (i) summarise the data supporting a role of the muscarinic system in schizophrenia, bipolar disorder and major depressive disorder, and (ii) give an overview of some of the new selective muscarinic ligands that are currently in development and try to address the issue of re-establishing appropriate central muscarinic function. PMID- 19816067 TI - Current and future applications of transcriptomics for discovery in CNS disease and injury. AB - The central nervous system (CNS) displays heterogeneity at regional, cellular and subcellular levels, making analysis of transcriptomic events accompanying neural injury particularly challenging. Microarray technology provides methods for elucidating global changes in neural gene expression and discovery of signalling pathways within this complex biological network. The lack of suitable and sufficient human CNS tissue along with its inherent variability means that diverse animal models of both multiple sclerosis and neurotrauma are vital for examining the pathophysiological changes accompanying neural injury resulting from disease or trauma. Gene expression profiling of these models is providing valuable information about mechanisms of damage, repair and regeneration and candidate treatments. In vitro models of neural injury are also proving useful, and transcriptomics is enhancing our understanding of the properties of neural stem cells with a view to their therapeutic application in neural repair. Thoughtful experimental design and analysis of microarray experiments is crucial for extracting biological meaning from the vast amount of data produced. In this review we discuss the current and emerging application of transcriptomics for the study of neural function in health, disease and injury. PMID- 19816068 TI - The influence of sex in non-small cell lung cancer. PMID- 19816069 TI - Microenvironmental influences and antigenic stimulation in chronic lymphocytic leukemia. PMID- 19816070 TI - Beyond platinum for metastatic and recurrent carcinoma of the cervix. PMID- 19816071 TI - Detection and prediction of breast cancer using double phase Tc-99m MIBI scintimammography in comparison with MRI. AB - BACKGROUND: The aim of this study was to investigate the incremental diagnostic value of quantified indices of scintimammography (SMM) additive to magnetic resonance imaging (MRI) for detection of breast cancer. PATIENTS AND METHODS: Double phase SMM (early: 10 min; delayed: 3 h) was performed in 239 highly suspected breast cancer patients. SMM was analyzed by visual and quantitative methods. Receiver operating characteristic curve (ROC) analyses were performed to determine the optimal visual grade, to calculate cut-off values of quantitative indices, and to evaluate incremental value. RESULTS: Sensitivity and specificity of visual analysis of SMM (V) were 86.4 and 100%, respectively. ROC analysis revealed 96.1% sensitivity and 58.8% specificity of contrast-enhanced MRI. The optimal lesion to non-lesion (L/N) ratios were 1.84 for early and 1.04 for delayed. When early L/N 1.84 was used as cut-off, sensitivity and specificity of SMM were 67.8 and 97.1%, respectively. When delayed L/N 1.04 was used, sensitivity and specificity were 88.3 and 55.9%, respectively. Among the various variables of double phase SMM and MRI, early L/N was the potent predictor for breast cancer. CONCLUSION: MRI and visual analysis early L/N showed similar diagnostic accuracy for breast cancer in the current study. Early L/N is a potential predictor for breast cancer. Further studies are needed to confirm these results and improve statistical accuracy. PMID- 19816073 TI - Lung cancer in the Canton of St. Gallen, Eastern Switzerland: sex-associated differences in smoking habits, disease presentation and survival. AB - BACKGROUND: The aim of this study was to assess sex-associated differences in lung cancer patients in Eastern Switzerland. METHODS: All 670 lung cancer patients referred to the cancer center in St. Gallen between January 2000 and December 2005 were retrospectively analyzed. We investigated sex-associated differences in age, smoking habits, histology, stage, treatment and survival. RESULTS: There were 474 (71%) men and 196 (29%) women with lung cancer. Mean age at the time of diagnosis was 64 years for women and 67 years for men (p = 0.01). Of the patients <55 years of age, 47 (24%) were women and only 65 (14%) were men. Men smoked significantly more than women (median pack-years: 50 vs. 30; p < 0.001). Of the heavy smokers (>40 pack-years), 278 (56%) were men and 68 (33%) were women. More men had squamous cell carcinoma (36%) than women (17%). Conversely, more women presented with adenocarcinoma (48%) than men (27%). No significant sex-associated differences were observed when analyzing first treatments received. Median overall survival was 10 months for both sexes. CONCLUSIONS: In Eastern Switzerland, women with lung cancer were younger, more likely to have smoked significantly less and more likely to have adenocarcinoma, compared to men with lung cancer. These findings are consistent with those found in other western populations. PMID- 19816072 TI - UFT plus oral folinic acid with alternating oral and intravenous vinorelbine in patients with metastatic breast cancer previously treated with anthracyclines and taxanes. AB - BACKGROUND: We retrospectively evaluated the activity and toxicity of uracil/tegafur (UFT) plus oral folinic acid in combination with vinorelbine (alternating intravenous (IV) on day 1 and oral on day 8) in patients with metastatic breast cancer. PATIENTS AND METHODS: Treatment consisted of IV vinorelbine 25 mg/m(2) on day 1 and 60 mg/m(2) orally on day 8, and oral UFT 300 mg/m(2) plus leucovorin 60 mg/m(2) on days 1-14 with 21 days interval. All patients were refractory to anthracyclines and taxanes. RESULTS: Partial response (PR) was observed in 3 (9.7%) and stable disease (SD) was observed in 13 (41.9%) patients. Total clinical benefit (PR + SD) of the combination was 51.6%. The median response duration was 3 (range 1-11.8) months. Median overall survival was 9.8 months (95% confidence interval (CI): 7.5-12.1), and median time to progression was 3.4 months (95% CI: 2.3-4.5). The most common toxicities were hematologic, including 4 (12.9%) cases of grade 3 neutropenia and 4 (12.9%) cases of grade 4 neutropenia. Grade 3 diarrhea was reported in 2 (3.2%) patients. 3 (9.7%) patients had hand-foot syndrome. Febrile neutropenia was observed in 1 patient. CONCLUSIONS: Although the combination is safe and convenient in clinical practice, it has only moderate activity in metastatic breast cancer patients. PMID- 19816074 TI - The relationship between cyclooxygenase-2, CD44v6, and nm23H1 in esophageal squamous cell carcinoma. AB - BACKGROUND: Esophageal cancer is the fourth most prevalent malignancy in China. The relationship between COX-2, CD44v6, and nm23H1 in esophageal squamous cell carcinoma (ESCC) remains unclear. MATERIAL AND METHODS: Expression of COX-2, CD44v6, and nm23H1 was examined, using the streptavidin-peroxidase method, in 82 ESCC and 30 normal esophageal mucosa (NEM) samples from the Shaanxi Province in China. RESULTS: The positive rates of COX-2, CD44v6, and nm23H1 were 73.2% (60/82), 64.6% (53/82), and 24.4% (31/82), respectively in ESCC, but 6.7% (2/30), 3.3% (1/30), and 90% (27/30), respectively in NEMs. There was a statistically significant difference between NEMs and ESCCs (p < 0.05). Expression of COX-2 showed a positive statistical correlation with expression of CD44v6 (r = 0.4732, p < 0.0001), and an inverse correlation with nm23H1 (r = -0.3226, p = 0.0035). Expression of COX-2, CD44v6, and nm23H1 had no significant correlation with gender or age (p > 0.05), but increased expression of COX-2 and CD44v6 showed statistical correlation with invasion and lymph node metastasis, respectively (p < 0.05). Decreased expression of nm23H1 was statistically correlated with lymph node metastasis (p = 0.0007) but had no correlation with invasion (p = 0.8221). CONCLUSIONS: This is the first report of a significant correlation between COX-2, CD44v6, and nm23H1 in ESCC. This knowledge might help us to further understand the molecular mechanisms of carcinogenesis and progression of ESCC. PMID- 19816075 TI - Toxic epidermal necrolysis related to pemetrexed and carboplatin with vitamin B12 and folic acid supplementation for advanced non-small cell lung cancer. AB - BACKGROUND: Pemetrexed is a multitargeted antifolate initially approved as a single agent for the second-line treatment of locally advanced or metastatic non small cell lung cancer and more recently in the first-line setting combined with cisplatin. The combination of pemetrexed with carboplatin has been tested in several phase II clinical trials showing interesting antitumour activity with mild toxicity. Supplementation with folic acid and vitamin B12 during treatment with pemetrexed is recommended to reduce potential haematological and gastrointestinal adverse events. CASE REPORT: A patient experienced cutaneous lesions including widespread erythema, epidermal detachment, and skin denudation, associated with deterioration of his general condition after the second cycle of this chemotherapy combination, which was clinically and histologically compatible with toxic epidermal necrolysis (Lyell's syndrome). Treatment with systemic steroids, antihistamines, and antibiotics led to resolution of the skin lesions and improvement of his general condition. CONCLUSION: To our knowledge, this is the second case reported in the literature of this type of suspected adverse drug reaction associated with a pemetrexed-based chemotherapy combination. PMID- 19816076 TI - Primary manifestation of small lymphocytic lymphoma in the prostate. AB - BACKGROUND: Infiltration of non-haematopoietic organs by small lymphocytic lymphoma/chronic lymphocytic leukaemia (SLL/CLL) is not unusual in late-stage disease and thus quite frequently encountered in post-mortem examinations. However, primary manifestation of SLL/CLL in the prostate is rarely diagnosed. PATIENTS AND METHODS: We report two cases of primary prostatic SLL/CLL, in one case in combination with prostate carcinoma, and discuss diagnostic pitfalls, pathophysiological mechanisms and therapeutic management, together with an overview of the literature. CONCLUSIONS: Lymphocytic infiltration of the prostate associated with obstructive symptoms is rare but can already occur in very early disease. Microscopically, SLL/CLL infiltration can be distinguished from chronic prostatitis by its pattern of infiltration and by immunohistochemistry. As the incidence of both SLL/CLL and prostatic carcinoma increases with age, composite tumours might occur more often in the future. PMID- 19816077 TI - Rectal intussusception due to re-recurrent ovarian fibrosarcoma. AB - BACKGROUND: Primary ovarian fibrosarcomas are very rare tumours associated with an extremely poor prognosis. Most patients relapse or die within 2 years. Because of the low incidence, it is difficult to identify prognostic factors or to establish treatment guidelines. CASE REPORT: In this report, we present a patient with the second relapse of an ovarian fibrosarcoma localised in the rectosigmoid. The tumour caused intussusception into the rectum, leading to large bowel obstruction. After complete resection of the tumour, the patient completely recovered. 28 months after primary diagnosis, follow-up did not show any signs of recurrent tumour disease. CONCLUSIONS: Reviewing the literature, it is recommended that all patients should primarily be operated on and generally be followed up closely. In patients with incomplete resection, palliative chemo- and/or radiotherapy is recommended. In our opinion, in patients with complete resection, adjuvant chemo- and/or radiotherapy should be considered. Only in patients with high-grade fibrosarcomas, adjuvant intra- or postoperative radiotherapy should be applied. PMID- 19816078 TI - Complete remission of metastatic and relapsed uterine cervical cancers using weekly administration of bevacizumab and paclitaxel/carboplatin. AB - BACKGROUND: Palliative therapy is usually employed for the treatment of metastatic or relapsed cervical cancer. Many agents including cisplatin have been used for fighting the tumor; however, the gold standard therapy has not yet been confirmed. CASE REPORT: Two cases of recurrent metastatic or refractory cervical cancer successfully treated with weekly administration of bevacizumab (2 mg/kg), paclitaxel (80 mg/m(2)), and carboplatin (area under the curve (AUC) = 2.0) are presented. 1 course of the therapy consisted of weekly paclitaxel/carboplatin on days 1, 8, and 15 and weekly bevacizumab on days 1, 8, 15, and 21, q28 days. Complete remission was observed after 3-4 courses of the therapy. Hematologic and non-hematologic toxicities higher than grade 3 were not observed during the chemotherapy. In both cases, there was no evidence of disease more than 10 months after the therapy. CONCLUSIONS: Weekly administration of bevacizumab and paclitaxel/carboplatin has potential activity in recurrent, metastatic, and refractory cervical carcinomas. These findings warrant further trials in such clinical settings. PMID- 19816079 TI - Incidence of febrile neutropenia and myelotoxicity of chemotherapy: a meta analysis of biosimilar G-CSF studies in breast cancer, lung cancer, and non Hodgkin's lymphoma. AB - BACKGROUND: The aim of this meta-analysis of 3 clinical studies, conducted with breast cancer, lung cancer, and non-Hodgkin's lymphoma patients, was to compare a new granulocyte colony-stimulating factor (G-CSF) biosimilar, XM02, with filgrastim in terms of its prophylactic effect on the development of febrile neutropenia (FN) during the first chemotherapy cycle in relation to the myelotoxic potency of the applied chemotherapy regimen. PATIENTS AND METHODS: Overall, 608 patients (363 under XM02 and 245 under filgrastim) were included in the meta-analysis. The majority of patients were allocated to the chemotherapy categories docetaxel-doxorubicin (45.4%) and cyclophosphamide-hydroxy daunomycin (adriamycin)-oncovin (vincristine)-prednisolone (CHOP)/platinum(Pt)-vinorelbine or Pt-vinblastine/ Pt-etoposide (43.1%); another 11.5% were allocated to the category Pt-gemcitabine/Pt-docetaxel or Pt-paclitaxel. RESULTS: FN in the XM02 and filgrastim groups was reported for 12.1 and 12.5% of patients, respectively, under docetaxeldoxorubicin, for 13.5 and 11.9% under CHOP/Pt-vinorelbine or Pt vinblastine/Pt-etoposide, and for 15.6 and 12.0% under Pt-gemcitabine/Pt docetaxel or Pt-paclitaxel. CONCLUSIONS: The incidence of FN in the first cycle of chemotherapy under primary G-CSF prophylaxis is low (in the range of 12-16%) and not directly correlated with the myelotoxic potency of the applied chemotherapy regimen. XM02 demonstrated to be non-inferior to filgrastim regarding the incidence of FN, irrespective of the myelotoxicity of the chemotherapy regimen. PMID- 19816080 TI - Cancer stem cells in solid tumors. AB - Carcinogenesis and tumor cell biology are very complex subjects. Recent findings on cancer stem cells may have a great impact on the understanding of the biology of malignancies and the development of future treatment approaches. Cancer stem cells are defined by their potential to self-renew and differentiate. Identification of cancer stem cells is possible by detecting expression of a certain combination of various cell surface markers and also by functional assays. The cancer stem cell concept was originally developed for hematological malignancies but is now becoming accepted for solid tumors as well. Emerging data indicates that cancer stem cells are responsible for the growth and spread of tumors. There is still a long way to go in characterizing cancer stem cells and understanding their role in carcinogenesis. Further studies should lead to a greater understanding of the biology of these cells with significant implications for diagnosis, treatment, and prevention of cancer. The recent studies regarding stem cell origin of various solid tumors will be discussed briefly in this review. PMID- 19816081 TI - Correlation of autophagy type in podocytes with histopathological diagnosis of IgA nephropathy. AB - OBJECTIVE: IgA nephropathy (IgA-N) frequently leads to progressive renal failure, thus estimation of the degree of progression is important for patient management. Autophagy is a mechanism that facilitates clearance of waste products to preserve renal function. The aim of this study was to assess autophagy in podocytes in children with progressive IgA-N at initial diagnosis by electron microscopy and investigate the relationship between the types of autophagy and severity of the disease. METHODS: Renal biopsies from 16 children with established progressive IgA-N were examined by light and transmission electron microscopy with reference to autophagy types in the podocytes and histopathological diagnosis of IgA-N. RESULTS: Two autophagy types were found. Type I rarely transformed to autophagic vacuoles and did not dissolve, thus possibly impairing cell function. However, type II frequently transformed to autophagosomes and autophagic vacuoles thus facilitating protein and lipid clearance. Of the 16 children studied, 8 (50%) with type I autophagy at initial diagnosis showed focal proliferative glomerulosclerosis (GN) of mild type (3 cases, 37.5%), mild/moderate type (2 cases, 25%) and moderate type (3 cases, 37.5%). In contrast, the remaining 8 children with type II autophagy at initial diagnosis showed focal proliferative GN of mild type in 7 (87.5%) and mild/moderate type in 1 (12.5%) case. CONCLUSION: In IgA-N children, the occurrence of type I autophagy is correlated with histopathologically more progressive disease, possibly reflecting a tendency to a poorer prognosis. PMID- 19816082 TI - Influence of insulin and muscle fiber type in nepsilon-(carboxymethyl)-lysine accumulation in soleus muscle of rats with streptozotocin-induced diabetes mellitus. AB - BACKGROUND: Nepsilon-(carboxymethyl)-lysine (CML) is an advanced glycation end product (AGE), the accumulation of which has been implicated in the etiology of diabetes complications. Skeletal muscle in diabetes demonstrates altered function, and increased accumulation of CML has been found in several fast-twitch muscles of diabetic animals. OBJECTIVE: This study aims to explore the accumulation of CML in soleus (a slow muscle) in diabetic animals, with and without insulin therapy. METHODS: Twenty-one rats were randomly divided into control and diabetes groups (DNI: diabetes without insulin; DI: diabetes with insulin; C: control). Diabetes was induced by intravenous administration of streptozotocin. At the end of the 12-week experimental period the soleus muscle was excised and snap frozen in liquid nitrogen. Muscle cross-sections were immunolabeled for CML. The number of CML-labeled muscle fibers was quantified; fibers were also evaluated for fiber types and cross-sectional areas. RESULTS: The percentage of myofibers immunolabeling for CML was highest in the DNI group (13.8 +/- 2.5%), lower in the DI group (5.4 +/- 1.1%) and lowest in the C group (2.1 +/- 0.6%). Statistical analysis revealed that AGE accumulation was significantly greater in the DNI group than in both C and DI groups (p = 0.0002). There was no significant difference between C and DI groups. In the DNI animals, AGE-positive myofibers showed a higher percentage of fast fiber types than did the AGE-negative fibers (49.5 +/- 6.9 vs. 13.7 +/- 1.5%, p = 0.002). No differences existed in cross-sectional areas between AGE-positive and AGE negative fibers within any group. CONCLUSION: The greatest accumulation of AGE was in the soleus of the DNI group, and was significantly less in the DI group. These findings may be linked to disordered glucose metabolism, increased oxidative stress and/or fiber type transformation in these muscles. PMID- 19816083 TI - BCL-6 oncoprotein in breast cancer: loss of expression in disease progression. AB - OBJECTIVE: To investigate the biological role of BCL-6 oncoprotein in breast cancer disease progression (recurrence and metastasis). METHODS: The series consisted of 93 consecutive female patients with primary breast cancer and median follow-up of 10 years. BCL-6 expression was assessed in vivo by immunohistochemistry and real-time PCR. Breast cancer cell lines and some metastasis-related genes (CXCR4, Itgbeta-3 and FLT-1) were also analysed by molecular techniques. Prognostic evaluation was performed by fitting a multivariate Cox regression model. RESULTS: BCL-6 immunoexpression was positive in 22 (23.7%) tumours and negative in 71 (76.3%). All axillary lymph node metastases of 47 node-positive patients were negative, including 12 cases showing BCL-6-positive primary tumours. Likewise, in 9 recurrence cases, BCL-6 expression was similar or decreased compared with primary tumours. No correlation between immunoexpression and gene expression of BCL-6 was observed. BCL-6 was significantly reduced both in derived metastases of a breast cancer cell line (M435) and when the latter was treated with a demethylation agent (5 azacytidine). However, BCL-6-transfected breast cancer cell lines expressed significantly higher levels of CXCR4, Itgbeta-3 and FLT-1. Co-expression of the 4 genes was found in 4 of 17 tumours evaluated, but lacking prognostic significance. BCL-6 oncoprotein revealed no significant influence on outcome. CONCLUSION: The results strongly suggest the loss of BCL-6 expression in breast cancer progression, which might be related with methylation status alterations of still unknown partner gene(s). PMID- 19816084 TI - Inflammatory infiltrates and neovessels are relevant sources of MMPs in abdominal aortic aneurysm wall. AB - OBJECTIVES: Abdominal aortic aneurysm (AAA) wall is characterized by degradation of extracellular matrix through matrix metalloproteinases (MMPs), chronic inflammatory cell infiltration and extensive neovascularization. So far, MMP expression within AAA wall in association with infiltrates and neovascularization has not yet been studied. METHODS: Vessel walls of 15 AAA patients and 8 organ donors were analyzed by immunohistochemistry for expression of various MMPs (MMP 1, -2, -3, -7, -8, -9, -12 and -13) in all cells located within the AAAs and correlated with infiltrates and neovascularization. RESULTS: Luminal endothelial cells (ECs) were positive for MMP-1, -3 and -9, ECs of mature neovessels were furthermore positive for MMP-2. Immature neovessels expressed all MMPs tested except for MMP-13. Aortic medial smooth muscle cells (SMCs) expressed MMP-1, -2, 3 and -9, SMCs of mature neovessels, only MMP-1, -3 and -9. Inflammatory infiltrates expressed all MMPs tested except for MMP-2, macrophages expressed all MMPs. Infiltrates were composed mainly of B cells (58.5 +/- 10.9%) and T lymphocytes (26.3 +/- 9.5%). Furthermore, significant inverse correlations were found between the amounts of inflammatory cells, neovessels and collagen/elastin content of the aortic vessel wall (r = +0.806/p < 0.001, r = -0.650/p = 0.012, r = -0.63/p < 0.015; respectively). CONCLUSION: Inflammatory infiltrates and invading neovessels are relevant sources of MMPs in the AAA wall and may substantially contribute to aneurysm wall instability. PMID- 19816085 TI - Effects of passive stretching on muscle injury and HSP expression during recovery after immobilization in rats. AB - OBJECTIVE: Stretching exercise is known to induce muscle hypertrophy and is implicated in the modulation of muscle fiber behavior. We aim to determine whether stretching exercise is protective against reloading-induced muscle damage in immobilized rat soleus muscle. METHODS: Rat hindlimbs in 54 eight-week-old male Wistar rats were immobilized by cast for 4 weeks, followed by reloading alone through normal ambulation in 24 (group NS) and after passive stretch in 25 rats (group S). Stretching exercise (30 min each day) lasted 6 days. To determine if passive stretching affects expression of heat shock proteins (HSPs) in rat soleus muscle during reloading following cast immobilization, the ratio of invading muscle fibers and HSPs expression were measured following cast removal. RESULTS: The ratio of invading muscle fibers increased during the first and second days of reloading in group NS. Compared with reloading alone, stretching exercise reduced invading muscle fibers at most time points following cast removal (group S). Additionally, expression of HSP25 and HSP72 increased with time during reloading only in the group without passive stretch (group NS). CONCLUSION: Following immobilization, in the rat soleus muscle passive stretching exercise protects against injury induced by reloading. Furthermore, the protection provided by passive stretch is independent of HSPs. PMID- 19816086 TI - IGF-I and IGF-binding proteins in articular exudates of children with post traumatic knee damage and juvenile idiopathic arthritis. AB - OBJECTIVE: IGF-I stimulates multiple functions of connective tissue cells and its activity is modulated by IGF-binding proteins (BPs). Some metalloproteinases are expected to modify IGF-I activity by digestion of IGF-BPs. It was decided to evaluate the concentration of IGF-I, IGF-BPs and the activity of gelatinases A and B in knee exudates of children with post-traumatic damage (PTD) and children with juvenile idiopathic arthritis (JIA) in comparison with those in the sera of the same patients. METHODS: ELISA (for IGF-I assay), polyacrylamine gel electrophoresis following Western immunoblotting (for IGF-I and IGF-BPs expression), and zymography (for gelatinase detection) were used. RESULTS: The knee exudates, especially those taken from patients with JIA, contained large amounts of IGF-I. The exudates of PTD and JIA patients contained some forms of IGF-BP-1 of molecular weight lower than those occurring in serum. Low expression BP-3 and high activity of gelatinase B were detected in the JIA exudates. CONCLUSIONS: The high gelatinase activities in exudates imply joint tissue damage. The cellular response to damage of this kind is an increase in IGF-I production, which stimulates repair processes. High proteolytic activities of gelatinase B in JIA patients may lower the amount of BP-3, possibly causing a relative decrease of IGF-I concentration and impairing the reparation processes stimulated by IGF-I. PMID- 19816087 TI - Antisense phosphorothioate oligodeoxynucleic acid for CD10 suppresses liver metastasis of colorectal cancer. AB - CD10 expression is associated with metastases of colorectal cancer (CRC). In the present study, we examined association of CD10 with liver metastasis of CRC cells to clarify the therapeutic significance of CD10. CD10-positive human colon cancer cell line, HT29 cells showed inhibition of growth, invasion and colony formation by treatment with CD10 antisense S-oligodeoxynucleotide (S-ODN). In the mouse liver metastasis mode, CD10 antisense S-ODN-treated HT29 cells made less embedded cells in the liver than control HT29 cells. Number and size of metastatic foci in nude mice liver were reduced in CD10 antisense S-ODN-treated HT29 cells. Treatment with CD10 antisense S-ODN decreased phosphorylation of ERK1/2 and EGFR in HT29 cells. Intraperitoneal administration of liposome-capsulated CD10 antisense S-ODN inhibited establishment of liver metastasis and growth of established metastasis in nude mice. These findings suggest that CD10 is associated substantially with liver metastasis of CRC cells and might be a molecular target of CRC treatment. PMID- 19816088 TI - Efficacy of repeated pretreatment with olopatadine hydrochloride on rhinitis induced by intranasal instillation of toluene-2,4-diisocyanate in rats. AB - It is well known that starting treatment for cedar pollinosis therapy with second generation antihistamines before the initial day of pollen scattering can relieve nasal symptom severity during the pollen season. Olopatadine hydrochloride (olopatadine) is an antiallergic agent with histamine H(1) receptor antagonistic action. We have evaluated the effects of repeated preadministration of olopatadine on the toluene-2,4-diisocyanate-induced rhinitis in rats. A single administration of olopatadine suppressed sneezing and the increases in histamine, nerve growth factor (NGF) and vascular endothelial growth factor (VEGF) production in nasal lavage fluid. When olopatadine was administered repeatedly once a day for 7 days before provocation, its inhibitory effects were enhanced compared to the effect of a single administration. Although the repeated administration of fexofenadine enhanced the inhibitory effects on sneezing, it did not inhibit the increases in NGF and VEGF production. These results show that the suppression of the increase in NGF and VEGF might partially be involved in the improvement of nasal allergy signs by the treatment with olopatadine. It is expected that the early treatment with olopatadine may achieve stable therapeutic effects. PMID- 19816089 TI - Role of lipoxygenases and the lipoxin A(4)/annexin 1 receptor in ischemia reperfusion-induced gastric mucosal damage in rats. AB - Rat gastric mucosal damage was induced by ischemia-reperfusion. The 5 lipoxygenase inhibitors MK886 and A63162, the 12-lipoxygenase inhibitor baicalein, the 15-lipoxygenase inhibitor PD146176 and the lipoxin (LX) A(4)/annexin 1 antagonist Boc1 increased mucosal damage in a dose-dependent manner. Low doses of these compounds, which have no effects on mucosal integrity, cause severe damage when combined with low doses of indomethacin, celecoxib or dexamethasone. 16,16-Dimethylprostaglandin (PG) E(2) and LXA(4) can replace each other in preventing mucosal injury induced by either cyclooxygenase or lipoxygenase inhibitors. The results suggest that not only cyclooxygenases, but also lipoxygenases have a role in limiting gastric mucosal damage during ischemia reperfusion. PMID- 19816090 TI - Neurotrophin receptors TrkA and TrkB in retinoblastoma are differentially expressed depending on cellular differentiation. AB - Retinoblastoma is the most common primary intraocular malignancy in children. With the progression of retinoblastoma, retinoblastoma cells lose their ability to differentiate. Regardless of many attempts to identify prognostic factors in retinoblastoma, further investigation for prognostic factors of retinoblastoma progression is still required because of the lack of sensitivity and specificity of these prognostic factors in predicting disease progression. We demonstrated that the differential expression of the neurotrophin receptors TrkA and TrkB is closely related to the differentiation of retinoblastoma cells. While retinoblastoma cells expressed TrkA as well as TrkB, their growth rates were not influenced by the addition of nerve growth factor to the culture medium. In experimental animal models of retinoblastoma, TrkA expression was primarily detected in more differentiated areas with high nm23 immunoreactivity whereas TrkB expression was apparent in more proliferative areas with high Ki67 immunoreactivity. With retinoic-acid-induced differentiation of retinoblastoma cells, TrkA expression significantly increased whereas TrkB significantly decreased. The differential expression of TrkA and TrkB with differentiation of retinoblastoma cells was mediated by extracellular-signal-regulated kinase 1/2 activation, which was confirmed by immunocytochemistry of TrkA. Therefore, our results suggest that the differential expression of TrkA and TrkB could be valuable as a therapeutic target, for instance using specific inhibitors. PMID- 19816091 TI - Promoter hypermethylation mediates downregulation of thiamine receptor SLC19A3 in gastric cancer. AB - As an important way to inactivate tumor suppressor genes (TSGs) during cancer development, promoter hypermethylation can be used to define novel TSGs and identify biomarkers for cancer diagnosis. SLC19A3 (solute carrier family 19, member 3) was found to be such a biomarker. SLC19A3 expression was downregulated in gastric cancer cell lines (71%, 5/7) and restored after pharmacological demethylation. Notably, hypermethylation of SLC19A3 promoter was detected in gastric cancer cell lines (57%, 4/7), primary gastric carcinoma tissues (51%, 52/101) and precancerous lesion (intestinal metaplasia) tissues (32%, 8/25). Exogenous SLC19A3 expression caused growth inhibition of gastric cancer cells. In summary, SLC19A3 was epigenetically downregulated in gastric cancer. Methylation of SLC19A3 promoter could be a novel biomarker for early gastric cancer development. PMID- 19816092 TI - Establishment and characterization of renal carcinoma cell lines from a Bhd gene mutant (Nihon) rat. AB - A germline insertion of a single nucleotide in the rat homologue of the human Birt-Hogg-Dube (BHD) gene gives rise to dominantly inherited renal cell carcinoma (RCC) in the Nihon rat model. In this study, we established 7 lines (NR cell lines NR22, 24, 32, 45, 49, 54 and 64) from an RCC found in a Nihon rat. All cell lines consisted mainly of round or polygonal cells arranged in a cobblestone-like growth pattern. Cells of NR cell lines had abundant cytoplasm and tight junctions as well as microvilli on electron microscopy and were positive for cytokeratin on immunocytochemistry. Cell lines NR22, 24 and 32 showed rapid growth, whereas the growth of the remaining lines was very slow. While the modal chromosome number of lines NR24, 45 and 54 was 42, the remaining lines exhibited aberrant modal numbers ranging from 70 to 96. All NR cell lines formed tumors at subcutaneous inoculation sites in nude mice, and tumors from lines NR54 and 64 developed pulmonary metastases. All NR cell lines had a germline mutation in the rat Bhd gene in the gene analysis. NR cell lines would prove valuable experimental tools for studies on unique functions of the Bhd gene and renal carcinogenesis. PMID- 19816093 TI - Ethylene, free radicals and the transition between stable states in plant morphology. AB - Treatment with hydrogen peroxide has notable effects in the morphology of the root apex in Arabidopsis seedlings. The result was described as consisting in two aspects: first, a reduction in curvature values in the root profile. Second, alterations in size and shape of the cells in the root cap. Cells of the root cap were smaller and had higher circularity index (Fig. 1). The results of peroxide treatment were similar to alterations in the root apex of ethylene insensitive mutants and wild-type seedlings treated with ethylene inhibitors. This brings new evidence in favour of the association between ethylene and hydrogen peroxide signalling that was recently demonstrated in stomatal cells. Notable changes in morphology under peroxide treatment were previously reported in other biological systems (Fig. 2). In the following paragraphs we make emphasis on the need of an accurate analysis of morphology. This aspect has not received the attention required in biology, a discipline dominated by functional analysis. We suggest that the observed morphological characteristics in the root apex treated with peroxide may be the manifestation of global processes of adaptation in the organism. Alternative forms of roots grown in water or in peroxide are stable situations representing different global configurations that may have other (genomic, physiological) traits associated. Each form represents a different mode of adaptation to environmental change. The accurate description of morphology in organisms, with particular emphasis in model systems, and their variations under stress, is needed to identify and understand the basis of genomic organization and plasticity. PMID- 19816094 TI - Plant intelligence: why, why not or where? AB - The concept of plant intelligence, as proposed by Anthony Trewavas, has raised considerable discussion. However, plant intelligence remains loosely defined; often it is either perceived as practically synonymous to Darwinian fitness, or reduced to a mere decorative metaphor. A more strict view can be taken, emphasizing necessary prerequisites such as memory and learning, which requires clarifying the definition of memory itself. To qualify as memories, traces of past events have to be not only stored, but also actively accessed. We propose a criterion for eliminating false candidates of possible plant intelligence phenomena in this stricter sense: an "intelligent" behavior must involve a component that can be approximated by a plausible algorithmic model involving recourse to stored information about past states of the individual or its environment. Re-evaluation of previously presented examples of plant intelligence shows that only some of them pass our test. PMID- 19816095 TI - The case against (-)-catechin involvement in allelopathy of Centaurea stoebe (spotted knapweed). AB - Proving allelopathic chemical interference is a daunting endeavor, in that production and movement of a phytotoxin from a donor plant to a receiving plant must be demonstrated in the substrate in which the plants grow, which is usually a complex soil matrix. The soil levels or soil flux levels of the compound generated by the donor must be proven to be sufficient to adversely affect the receiving plant. Reports of (-)-catechin to be the novel weapon used by Centaurea stoebe (spotted knapweed) to invade new territories are not supported by the paper featured in this Addendum, nor by papers produced by two other laboratories. These papers find that (-)-catechin levels in soil in which C. stoebe grows are orders of magnitude below levels that cause only minor growth effects on reported sensitive species. Furthermore, the claim that (-)-catechin acts as a phytotoxin through causing oxidative damage is refuted by the fact that the molecule is a strong antioxidant and is quickly degraded by extracellular root enzymes. PMID- 19816096 TI - Covering common ground: F-actin-dependent transport of plant viral protein inclusions reveals a novel mechanism for movement utilized by unrelated viral proteins. AB - Plant viruses are composed of diverse genomes (e.g., RNA or DNA) encoding proteins that vary widely in sequence. It is becoming clear, however, that some apparently unrelated viral proteins have similar functions. The P6 protein encoded by Cauliflower mosaic virus (CaMV) and the 126-kDa protein encoded by Tobacco mosaic virus (TMV) are examples of this convergence in protein function. Although having no apparent sequence similarity, both proteins are pathogenicity determinants during infection, are components of novel intracellular cytoplasmic inclusions and suppress RNA silencing. Here we review our recent results demonstrating an additional novel convergent activity between these proteins: both proteins traffic along the actin cytoskeleton (microfilaments). We also discuss results showing a unique property of the P6 protein: a non-mobile strong association with microtubules. Lastly, we discuss the potential mechanism by which the P6 and 126-kDa proteins traffic along microfilaments. We provide new results suggesting that actin filament polymerization-driven movement does not support 126-kDa protein transport, thus leading to a focus on myosins as the driving force for this movement. PMID- 19816097 TI - Isolation of high salinity stress tolerant genes from Pisum sativum by random overexpression in Escherichia coli and their functional validation. AB - Salinity stress is one of the major factors which reduce crop plants growth and productivity resulting in significant economic losses worldwide. Therefore, it would be fruitful to isolate and functionally identify new salinity stress induced genes for understanding the mechanism and developing salinity stress tolerant plants. Based on functional gene screening assay, we have isolated few salinity tolerant genes out of one million Escherichia coli (SOLR) transformants containing pea cDNAs. Sequence analysis of three of these genes revealed homology to Ribosomal-L30E (RPL30E), Chlorophyll-a/b-binding protein (Chla/bBP) and FIDDLEHEAD (FDH). The salinity tolerance of these genes in bacteria was further confirmed by using another strain of E. coli (DH5alpha) transformants. The homology based computational modeling of these proteins suggested the high degree of conservation with the conserved domains of their homologous partners. The reverse transcriptase polymerase chain reaction (RT-PCR) analysis showed that the expression of these cDNAs (except the FDH) was upregulated in pea plants in response to NaCl stress. We observed that there was no significant effect of Li(+) ion on the expression level of these genes, while an increase in response to K(+) ion was observed. Overall, this study provides an evidence for a novel function of these genes in high salinity stress tolerance. The PsFDH showed constitutive expression in planta suggesting that it can be used as constitutively expressed marker gene for salinity stress tolerance in plants. This study brings new direction in identifying novel function of unidentified genes in abiotic stress tolerance without previous knowledge of the genome sequence. PMID- 19816098 TI - The RHA2a-interacting proteins ANAC019 and ANAC055 may play a dual role in regulating ABA response and jasmonate response. AB - Jasmonate- and ABA-mediated signalings are involved in the activation of defense responses of plants to biotic and abiotic stresses. Accumulating evidence has suggested the existence of comprehensive synergistic or antagonistic cross-talks between these two signaling pathways. However, relatively little is known about how these cross-talks are executed at the molecular level. Our recent works have implied that, ANAC019 and ANAC055, two highly related NAC family transcription factors in Arabidopsis, may play a dual role in regulating jasmonate response and ABA response. PMID- 19816099 TI - Maintenance of stress related transcripts in tolerant cultivar at a level higher than sensitive one appears to be a conserved salinity response among plants. AB - Response of plants towards salinity is multigenic in nature with its various components playing diverse roles in stress perception, relay or response. For the purpose of dissecting the genetic determinants of salinity response in crops, the family Brassicaceae presents an excellent model since significant inter-and intra specific variations have been reported for salinity tolerance. Using these intraspecific variations of Brassica, we show that one of the possible mechanism by which a genotype is able to exhibit tolerance better than another is by keeping the basal levels of stress responsive transcripts higher than the sensitive genotype. This is quite reflected when we analyze members of a specific pathway such as SOS pathway or even when we extend the analysis to a range of molecules including those playing important role in stress perception, signal transduction or stress response. However, these investigations need to be extended to genome level transcript analysis to further validate the hypothesis of "well preparedness" in tolerant genotypes and we propose the suitability of Brassica genotypes for this endevours. PMID- 19816100 TI - The missing link?: Arabidopsis SPCH is a MAPK specificity factor that controls entry into the stomatal lineage. AB - Mitogen activated protein kinase (MAPK) signaling modules that incorporate AtMPK3 and AtMPK6 control critical aspects of Arabidopsis biology including stress responses, development, cell division and cell death. Arabidopsis stomatal development is negatively regulated by the YDA-MKK4/5-MPK3/6 MAPK module and follows a three step pathway of asymmetric and symmetric divisions followed by terminal differentiation. We have identified the bHLH transcription factor SPCH, which controls entry into the stomatal lineage as a substrate of AtMPK3 and AtMPK6. These findings suggest that SPCH activity may be directly affected by environmental conditions to enable the plant to modify stomatal development in response to suboptimal climates. PMID- 19816101 TI - FZR2/CCS52A1 mediated endoreduplication in Arabidopsis development. AB - Plant organogenesis generally involves three basic processes: cell division, cell expansion and cell differentiation. Endoreduplication, a process of genome replication without intervening mitosis, often occurs during cell expansion and cell differentiation. The switch from the mitotic cell cycle to the endocycle, however, is still poorly understood in plants. We have recently demonstrated that FIZZY-RELATED2 (FZR2) is a factor controlling endoreduplication in Arabidopsis. fzr2 mutants lacked gross morphological defects but showed a general decrease of endoploidy level in trichomes and other leaf cells, while expression of FZR2 under constitutive or tissue specific promoters induced extra or ectopic endoreduplication in all tissues examined. We also showed that decrease of leaf cell size in fzr2 mutants could be compensated by increased cell proliferation. In this addendum, we discuss additional phenotypes of FZR2 misexpression, including apparent mosaic leaf sectors in which local cell overexpansion due to 35S::FZR2 appears to be compensated by reduced cell expansion in neighboring tissues. PMID- 19816102 TI - Assessing fungal root colonization for plant improvement. AB - Fungal endophytes display a broad range of symbiotic interactions with their host plants. Current studies on their biology, diversity and benefits are unravelling their high relevance on plant adaptation to environmental stresses. Implementation of such properties may open new perspectives in agriculture and forestry. We aim to exploit the endophytic capacities of the fungal species Fusarium equiseti, a naturally occurring root endophyte which has shown antagonism to plant pathogens, and Pochonia chlamydosporia, a nematophagous fungus with putative endophytic behavior, for plant protection and adaptation to biotic and abiotic stress. A real-time PCR protocol for quantification of the fungal population, together with Agrobacterium-mediated genetic transformation with the GFP gene for confocal microscopy analyses, were designed and applied to assess endophytic development of both these fungal species. Although quantification of both F. equiseti and P. chlamydosporia yielded similar degrees of root colonization, microscopical observations demonstrated differences in infection and development patterns. Furthermore, we found evidences of plant response against endophyte colonization, supporting a balanced antagonism between the endophyte virulence and the plant defenses. Optimization and application of the methodologies presented herein will allow elucidation of beneficial interactions among these endophytes and their host plants. PMID- 19816103 TI - CDPK1, a calcium-dependent protein kinase, regulates transcriptional activator RSG in response to gibberellins. AB - The homeostasis of gibberellins (GAs) is maintained by negative-feedback regulation in plant cells. REPRESSION OF SHOOT GROWTH (RSG) is a transcriptional activator with a basic Leu zipper domain suggested to contribute GA feedback regulation by the transcriptional regulation of genes encoding GA biosynthetic enzymes. The 14-3-3 signaling proteins negatively regulate RSG by sequestering it in the cytoplasm in response to GAs. The phosphorylation on Ser-114 of RSG is essential for 14-3-3 binding of RSG; however, the kinase that catalyzes the reaction is unknown. Recently a Ca(2+)-dependent protein kinase (CDPK) was identified as an RSG kinase that promotes 14-3-3 binding of RSG by phosphorylation of the Ser-114 of RSG. Our results suggest that CDPK decodes the Ca(2+) signal produced by GAs and regulates the intracellular localization of RSG in plant cells. PMID- 19816105 TI - Amplification of phytochrome induced morphogenesis in plants by the cryptic red light signal (CRS). AB - The regulation of endogenous levels of ascorbic acid in soybean by far-red absorbing form of phytochrome (Pfr) and by cryptic red light signal (CRS) was studied. Cryptic red light signal is produced by red light pre-irradiation of a photoreceptor other than far-red absorbing form of phytochrome (Pfr) and CRS amplifies the action of phytochrome. The endogenous level of ascorbic acid levels enhanced by phytochrome was amplified by CRS. The lifetime of CRS was from 0 to 2 h and the peak of enhancement of ascorbic acid due to CRS was between 16 to 24 h of dark incubation after the end of the treatment. CRS was found to be ineffective on UV-B enhanced endogenous levels of ascorbic acid. PMID- 19816104 TI - Soluble sugars--metabolism, sensing and abiotic stress: a complex network in the life of plants. AB - Plants are autotrophic and photosynthetic organisms that both produce and consume sugars. Soluble sugars are highly sensitive to environmental stresses, which act on the supply of carbohydrates from source organs to sink ones. Sucrose and hexoses both play dual functions in gene regulation as exemplified by the upregulation of growth-related genes and downregulation of stress-related genes. Although coordinately regulated by sugars, these growth- and stress-related genes are upregulated or downregulated through HXK-dependent and/or HXK-independent pathways. Sucrose-non-fermenting-1- (SNF1-) related protein pathway, analogue to the protein kinase (SNF-) yeast-signalling pathway, seems also involved in sugar sensing and transduction in plants. However, even if plants share with yeast some elements involved in sugar sensing, several aspects of sugar perception are likely to be peculiar to higher plants. In this paper, we have reviewed recent evidences how plants sense and respond to environmental factors through sugar sensing mechanisms. However, we think that forward and reverse genetic analysis in combination with expression profiling must be continued to uncover many signalling components, and a full biochemical characterization of the signalling complexes will be required to determine specificity and cross-talk in abiotic stress signalling pathways. PMID- 19816106 TI - A novel plant cysteine-rich peptide family conferring cadmium tolerance to yeast and plants. AB - We have identified a novel cDNA clone, termed DcCDT1, from Digitaria ciliaris, that confers cadmium (Cd)-tolerance to yeast (Saccharomyces cerevisiae). The gene encodes a predicted peptide of 55 amino acid residues of which 15 (27.3%) are cysteine residues. We found that monocotyledonous plants possess multiple DcCDT1 homologues, for example rice contains five DcCDT1 homologues (designated OsCDT1~5), whereas dicotyledonous plants, including Arabidopsis thaliana, Brassica rapa, poplar (Populus tremula x Populus alba) and Picea sitchensis, appear to possess only a single homologue. GFP fusion experiments demonstrate that DcCDT1 and OsCDT1 are targeted to both the plant cytoplasmic membranes and cell walls. Constitutive expression of DcCDT1 or OsCDT1 confers Cd-tolerance to transgenic A. thaliana plants by lowering the accumulation of Cd in the cells. The functions of the DcCDT1 family members are discussed in the light of these findings. PMID- 19816107 TI - Heme-independent soluble and membrane-associated peroxidase activity of a Zea mays annexin preparation. AB - Annexins are cytosolic proteins capable of reversible, Ca(2+)-dependent membrane binding or insertion. Animal annexins form and regulate Ca(2+)-permeable ion channels and may therefore participate in signaling. Zea mays (maize) annexins (ZmANN33 and ZmANN35) have recently been shown to form a Ca(2+)-permeable conductance in planar lipid bilayers and also exhibit in vitro peroxidase activity. Peroxidases form a superfamily of intra- or extracellular heme containing enzymes that use H(2)O(2) as the electron acceptor in a number of oxidative reactions. Maize annexin peroxidase activity appears independent of heme and persists after membrane association, the latter suggesting a role in reactive oxygen species signaling. PMID- 19816108 TI - Rice OsSIPK: a central component of ozone-triggered physiological responses. AB - The OsSIPK expression is transcriptionally regulated in time and space by diverse environmental stresses and phytohormones. Rice OsSIPK and its orthologs in other plants are highly conserved and appear to have overlapping physiological responses. Given our interest in understanding the signaling and metabolic pathways responsible for environmental factors, we briefly discuss the role of OsSIPK in ozone-triggered physiological responses, particularly in rice. We also provide evidence on tight correlation between ozone-induced OsSIPK expression and ethylene production. PMID- 19816109 TI - The CaMV 35S promoter has a weak expression activity in dark grown tissues of moss Physcomitrella patens. AB - The constitutive Cauliflower Mosaic Virus 35S promoter (CaMV 35S) is widely used as a tool to express recombinant proteins in plants, but with different success. We previously showed that the expression of an F-actin marker, GFP-talin, in Physcomitrella patens using the CaMV 35S promoter failed to homogenously label moss tissues. Here, we show a significant diminution of the GFP fluorescence in dark grown old moss cells and complete lack of labelling in newly differentiated cells. Furthermore, we demonstrate that stable moss lines harbouring a resistance cassette driven by the CaMV 35S are unable to grow in darkness in the presence of the antibiotic. In contrast to the CaMV 35S, the heat inducible promoter, hsp17.3B showed uniform expression pattern in all cells and tissues following a mild heat shock. PMID- 19816111 TI - Roles of DLT in fine modulation on brassinosteroid response in rice. AB - Accumulated evidences suggest that brassinosteroid (BR) response is fine-tuned by multiple strategies. Recently, a GRAS protein, DLT, was identified to be involved in rice BR signaling. Transcription analysis of most known BR-related genes, including biosynthetic genes, signaling genes and also several downstream BR responsive genes, reveals DLT has extensive effects on BR-related genes expression, indicating its important role in modulating BR response. By promoter and protein sequence analysis, a complicated and subtle network was proposed that DLT may be involved in fine-modulating BR response at both transcriptional and protein levels. This provides new insights into BR signaling pathway and also gives a clear direction for the future work. PMID- 19816110 TI - Circumnutation as a visible plant action and reaction: physiological, cellular and molecular basis for circumnutations. AB - Circumnutation is a helical organ movement widespread among plants. It is variable due to a different magnitude of trajectory (amplitude) outlined by the organ tip, duration of one cycle (period), circular, elliptical, pendulum-like or irregular shape and clock- and counterclockwise direction of rotation. Some of those movement parameters are regulated by circadian clock and show daily and infradian rhythms. Circumnutation is influenced by light, temperature, chemicals and can depend on organ morphology. The diversity of this phenomenon is easier to see now that the digital time-lapse video method is developing fast. Whether circumnutation is an endogenous action, a reaction to exogenous stimuli or has a combined character has been discussed for a long time. Similarly, the relationship between growth and circumnutation is still unclear. In the mechanism of circumnutation, epidermal and endodermal cells as well as plasmodesmata, plasma membrane, ions (Ca(2+), K(+) and Cl(-)), ion channels and the proton pump (H(+)ATPase) are engaged. Based on these data, the hypothetical electrophysiological model of the circumnutation mechanism has been proposed here. In the recent circumnutation studies, gravitropic, auxin, clock and phytochrome mutants are used and new functions of circumnutation in plants' life have been investigated and described. PMID- 19816112 TI - Role of nitric oxide in regulating stomatal apertures. AB - During stomatal closure, nitric oxide (NO) operates as one of the key intermediates in the complex, abscisic acid (ABA)-mediated, guard cell signaling network that regulates this process. However, data concerning the role of NO in stomatal closure that occurs in turgid vs. dehydrated plants is limited. The data presented demonstrate that, while there is a requirement for NO during the ABA induced stomatal closure of turgid leaves, such a requirement does not exist for ABA-enhanced stomatal closure observed to occur during conditions of rapid dehydration. The data also indicate that the ABA signaling pathway must be both functional and to some degree activated for guard cell NO signaling to occur. These observations are in line with the idea that the effects of NO in guard cells are mediated via a Ca(2+)-dependent rather than a Ca(2+)-independent ABA signaling pathway. It appears that there is a role for NO in the fine tuning of the stomatal apertures of turgid leaves that occurs in response to fluctuations in the prevailing environment. PMID- 19816113 TI - The fate and the role of mitochondria in Fe-deficient roots of strategy I plants. AB - In well aerated soils, iron exists, mainly as scarcely soluble oxides and oxi hydroxides and, therefore, not freely available to plants uptake, notwithstanding its abundance. Multifaceted strategies involving reductase activities, proton processes, specialized storage proteins, and other, act in concert to mobilize iron from the environment, to take it up and to distribute it inside the plant. Because of its fundamental role in plant productivity several questions concerning homeostasis of iron in plants are currently a matter of intense debate. We discuss some recent studies on Strategy I responses in dicotyledonous plants focusing on metabolic change induced by iron deficiency, mainly concerning the involvement of mitochondria. PMID- 19816114 TI - Negative effects of desiccation on the protein sorting and post-translational modification. AB - Bryophytes as the first land plants are believed to have colonized the land from a fresh water origin, requiring adaptive mechanisms that survival of dehydration. Physcomitrella patens is such a non-vascular bryophyte and shows rare desiccation tolerance in its vegetative tissues. Previous studies showed that during the course of dehydration, several related processes are set in motion: plasmolysis, chloroplast remodeling and microtubule depolymerization. And proteomic alteration supported the cellular structural changes in respond to desiccation stress. In this addendum, we report that Golgi bodies are absent and adaptor protein complex AP-1 large subunit is downregulated during the course of dehydration. Those phenomena may be adverse in protein posttranslational modification, protein sorting and cell walls synthesis under the desiccation condition. PMID- 19816115 TI - Alternative polyadenylation: a mechanism maximizing transcriptome diversity in higher eukaryotes. AB - Based on comparative genome analyses, the increases in protein-coding gene number could not account for the increases of morphological and behavioral complexity of higher eukaryotes. Transcriptional regulations, alternative splicing and the involvement of non-coding RNA in gene expression regulations have been credited for the drastic increase of transcriptome complexity. However, an emerging theme of another mechanism that contributes to the formation of alternative mRNA 3' ends is alternative polyadenylation (APA). First, recent studies indicated that APA is a wide spread phenomenon across the transcriptomes of higher eukaryotes and being regulated by developmental and environmental cues. Secondly, our characterization of the Arabidopsis polyadenylation factors suggested that plant polyadenylation has also evolved to regulate the expression of specific genes by means of APA and therefore the specific biological functions. Finally, Phylogenetic analyses of eukaryotic polyadenylation factors from several organisms revealed that the number of polyadenylation factors tends to increase in higher eukaryotes, which provides the potential for their functional differentiation in regulating gene expression through APA. Based on above evidence, we, thus, hypothesize that APA, serving as an additional mechanism, contributes to the complexity of higher eukaryotes. PMID- 19816116 TI - Nitric oxide and nitrite are likely mediators of pollen interactions. AB - The ability of plants to produce nitric oxide (NO) is now well recognised. In plants, NO is involved in the control of organ development and in regulating some of their physiological functions. We have recently shown that pollen generates NO in a constitutive manner and have measured both intra- and extracellular production of this radical. Furthermore, we have shown that nitrite accumulates in the media surrounding the pollen and have suggested that the generation of these signaling molecules may be important for the normal interaction between the pollen grain and the stigma on which it alights. However, pollen grains inevitably come into contact with other tissues, including those of animals and it is likely that the NO produced will influence the behavior of the cells associated with these tissues. Such non-animal-derived, NO-mediated effects on mammalian cells may not be restricted to pollen and plant debris and fungal spore derived NO may elicit similar effects. PMID- 19816117 TI - Molecular cloning of acetylcholinesterase gene from Salicornia europaea L. PMID- 19816118 TI - Improving rhizome yield and quality of Paris polyphylla through gibberellic acid induced retardation of senescence of aerial parts. AB - Senescence in perennials has not been extensively studied compared to that in annual plants, and the effects of delaying senescence on plant biomass and metabolic features in herbaceous perennials has also been poorly reported. We recently examined the effects of gibberellin A(3) (GA(3)) on senescence of aerial parts of Paris polyphylla, which characterize the metabolic changes associated with senescence, and found antagonistic effects of GA and abscisic acid during this process. Rhizome yield and quality (saponin content) of Paris polyphylla were both improved by GA(3)-induced retardation of senescence. We propose that GA(3)-induced retardation of senescence increases green leaf area and prolongs the duration of photosynthesis, leading to increased rhizome yield. The increased saponin accumulation in GA(3)-treated plants may be explained by the longer growth phase and ensuing increased environmental stress. PMID- 19816119 TI - DOR: a link between an F-box protein and guard cell ABA signaling. AB - Guard cells are a model system for studying signal transduction. F-box proteins, representing one of the largest gene families in Arabidopsis, have been shown to be involved in many developmental and physiological processes, including stress responses. However, it is unclear if there is a direct link between an F-box protein and the guard cell ABA signaling. DOR is a guard cell-preferential F-box protein, and our results suggested that it likely forms two negative feedback regulatory loops for the ABA-induced stomatal closure under drought conditions in Arabidopsis. These findings have a potential impact on genetically modifying drought stress responses in plants. PMID- 19816120 TI - Cloning of 9-cis-epoxycarotenoid dioxygenase (NCED) gene and the role of ABA on fruit ripening. AB - In order to understand more details about the role of abscisic acid (ABA) in fruit ripening and senescence, six 740 bp cDNAs (LeNCED1, LeNCED2, PpNCED1, VVNCED1, DKNCED1 and CMNCED1) which encode 9-cis-epoxycarotenoid dioxygenase (NCED) as a key enzyme in ABA biosynthesis, were cloned from fruits of tomato, peach, grape, persimmon and melon using an RT-PCR approach. A Blast homology search revealed a similarity of amino acid 85.76% between the NCEDs. A relationship between ABA and ethylene during ripening was also investigated. At the mature green stage, exogenous ABA treatment increased ABA content in flesh, and promoting ethylene synthesis and fruit ripening, while treatment with nordihydroguaiaretic acid (NDGA), inhibited them, delayed fruit ripening and softening. However, ABA inhibited the ethylene synthesis obviously while NDGA promoted them when treated the immature fruit with these chemicals. At the breaker, NDGA treatment cannot block ABA accumulation and ethylene synthesis. Based on the results obtained in this study, it was concluded that ABA plays different role in ethylene synthesis system in different stages of tomato fruit ripening. PMID- 19816121 TI - Plant neurobiology: From stimulus perception to adaptive behavior of plants, via integrated chemical and electrical signaling. PMID- 19816122 TI - The Arabidopsis peroxisome division mutant pdd2 is defective in the DYNAMIN RELATED PROTEIN3A (DRP3A) gene. PMID- 19816123 TI - Ant-plant mutualisms should be viewed as symbiotic communities. PMID- 19816124 TI - Promoting gene expression in plants by permissive histone lysine methylation. AB - Plants utilize sophisticated epigenetic regulatory mechanisms to coordinate changes in gene expression during development and in response to environmental stimuli. Epigenetics refers to the modification of DNA and chromatin associated proteins, which affect gene expression and cell function, without changing the DNA sequence. Such modifications are inherited through mitosis, and in rare instances through meiosis, although it can be reversible and thus regulatory. Epigenetic modifications are controlled by groups of proteins, such as the family of histone lysine methytransferases (HKMTs). The catalytic core known as the SET domain encodes HKMT activity and either promotes or represses gene expression. A large family of SET domain proteins is present in Arabidopsis where there is growing evidence that two classes of these genes are involved in promoting gene expression in a diverse range of developmental processes. This review will focus on the function of these two classes and the processes that they control, highlighting the huge potential this regulatory mechanism has in plants. PMID- 19816125 TI - Biosynthesis of salicylic acid in plants. AB - Salicylic acid (SA) is an important signal molecule in plants. Two pathways of SA biosynthesis have been proposed in plants. Biochemical studies using isotope feeding have suggested that plants synthesize SA from cinnamate produced by the activity of phenylalanine ammonia lyase (PAL). Silencing of PAL genes in tobacco or chemical inhibition of PAL activity in Arabidopsis, cucumber and potato reduces pathogen-induced SA accumulation. Genetic studies, on the other hand, indicate that the bulk of SA is produced from isochorismate. In bacteria, SA is synthesized from chorismate through two reactions catalyzed by isochorismate synthase (ICS) and isochorismate pyruvate lyase (IPL). Arabidopsis contains two ICS genes but has no gene encoding proteins similar to the bacterial IPL. Thus, how SA is synthesized in plants is not fully elucidated. Two recently identified Arabidopsis genes, PBS3 and EPS1, are important for pathogen-induced SA accumulation. PBS3 encodes a member of the acyl-adenylate/thioester-forming enzyme family and EPS1 encodes a member of the BAHD acyltransferase superfamily. PBS3 and EPS1 may be directly involved in the synthesis of an important precursor or regulatory molecule for SA biosynthesis. The pathways and regulation of SA biosynthesis in plants may be more complicated than previously thought. PMID- 19816126 TI - Callose synthesis in higher plants. AB - Callose is a polysaccharide in the form of beta-1,3-glucan with some beta-1,6 branches and it exists in the cell walls of a wide variety of higher plants. Callose plays important roles during a variety of processes in plant development and/or in response to multiple biotic and abiotic stresses. It is now generally believed that callose is produced by callose synthases and that it is degraded by beta-1,3-glucanases. Despite the importance of callose in plants, we have only recently begun to elucidate the molecular mechanism of its synthesis. Molecular and genetic studies in Arabidopsis have identified a set of genes that are involved in the biosynthesis and degradation of callose. In this mini-review, we highlight recent progress in understanding callose biosynthesis and degradation and discuss the future challenges of unraveling the mechanism(s) by which callose synthase operate. PMID- 19816127 TI - Cortex proliferation: simple phenotype, complex regulatory mechanisms. PMID- 19816128 TI - The importance of floral signals in the establishment of plant-ant mutualisms. PMID- 19816129 TI - Comparison of salt-responsive gene regulation in rice and in the salt-tolerant Festuca rubra ssp. litoralis. PMID- 19816130 TI - The rhizosphere bacterium Variovorax paradoxus 5C-2 containing ACC deaminase does not increase systemic ABA signaling in maize (Zea mays L.). PMID- 19816131 TI - Grapevine MLO candidates required for powdery mildew pathogenicity? PMID- 19816132 TI - The LysM receptor kinase CERK1 mediates bacterial perception in Arabidopsis. PMID- 19816133 TI - Cytosolic alkalinization is a common and early messenger preceding the production of ROS and NO during stomatal closure by variable signals, including abscisic acid, methyl jasmonate and chitosan. PMID- 19816134 TI - Retraction of : "The multilevel and dynamic interplay between plant and pathogen". PMID- 19816135 TI - The Arabidopsis membrane-bound transcription factor AtbZIP60 is a novel plant specific endoplasmic reticulum stress transducer. PMID- 19816136 TI - Boron deficiency: how does the defect in cell wall damage the cells? PMID- 19816137 TI - Mullerian mimicry in aposematic spiny plants. AB - Mullerian mimicry is common in aposematic animals but till recently, like other aspects of plant aposematism was almost unknown. Many thorny, spiny and prickly plants are considered aposematic because their sharp defensive structures are colorful and conspicuous. Many of these spiny plant species (e.g., cacti and Agave in North American deserts; Aloe, Euphorbia and acacias with white thorns in Africa; spiny plants in Ohio; and spiny members of the Asteraceae in the Mediterranean basin) have overlapping territories, and also similar patterns of conspicuous coloration, and suffer from the evolutionary pressure of grazing by the same large herbivores. I propose that many of these species form Mullerian mimicry rings. PMID- 19816138 TI - Quantitative proteomics identifies oxidant-induced, AtMPK6-dependent changes in Arabidopsis thaliana protein profiles. AB - In Arabidopsis thaliana, oxidant-induced signalling has been shown to utilize the mitogen-activated protein kinase (MAPK), AtMPK6. To identify proteins whose accumulation is altered by ozone in an AtMPK6-dependent manner we employed isotope-coded affinity tagging (ICAT) technology to investigate the impact of AtMPK6-suppression on the protein profiles in Arabidopsis both before (air control) and during continuous ozone (O(3)) fumigation (500 nL L(-1) for 8 h). Among the 150 proteins positively identified and quantified in the O(3)-treated plants, we identified thirteen proteins whose abundance was greater in the AtMPK6 suppressed genotype than in wild-type (WT). These include the antioxidant proteins, monodehydroascorbate reductase, peroxiredoxin Q, and glutathione reductase. A further eighteen proteins were identified whose abundance was lower in the ozone-treated AtMPK6-suppressed line relative to ozone-exposed WT plants. These predominantly comprised proteins involved in carbohydrate-, energy-, and amino acid metabolism, and tetrapyrrole biosynthesis. In control plants, five proteins increased, and nine proteins decreased in abundance in the AtMPK6 suppressed genotype compared to that of the WT, reflecting changes in the protein composition of plants that have AtMPK6 constitutively suppressed. Since a number of these proteins are part of the redox response pathway, and loss of AtMPK6 renders Arabidopsis more susceptible to oxidative stress, we propose that AtMPK6 plays a key role in the plant's overall ability to manage oxidative stress. PMID- 19816139 TI - Aphid transmission of cauliflower mosaic virus: the role of the host plant. PMID- 19816140 TI - Evolution of CLE signaling: origins of the CLV1 and SOL2/CRN receptor diversity. AB - The shoot apical meristem is maintained by the intercellular factor, CLV3, a dodecapeptide in Arabidopsis. CLV3 belongs to the CLE family and putative CLE genes have been found in various plants, even in the moss Physcomitrella patens. Here, we report that a pteridophyte, Selaginella moelendorffii, also has 15 putative CLE genes in its genome. On the other hand, CLV1 is reported to function as a receptor for the CLV3 peptide, and other CLE peptides might be recognized by CLV1 homologues in various plants. Recent genetic studies of the crn and sol2 mutants of Arabidopsis have revealed that SOL2/CRN encodes a receptor-like kinase protein. SOL2/CRN functions together with CLV2 independently of CLV1 in the CLE signaling pathway. Phylogenetic analysis of CLV1, CLV2 and SOL2/CRN revealed that Arabidopsis, rice, Populus trichocarpa and Vitis vinifera have one copy of the SOL2/CRN and CLV2 homologues, and Selaginella moelendorffii and Physcomitrella patens have no homologues. In contrast, a number of CLV1 homologues were identified in the genomic databases of Arabidopsis, rice, Populus trichocarpa, Vitis vinifera, and even a pteridophyte, Selaginella moelendorffii, and a moss, Physcomitrella patens. These results indicate that CLV1 and its homologues play multiple roles in plant development and environmental responses, whereas SOL2/CRN and CLV2 have more specific roles in vascular plants. PMID- 19816142 TI - Identification of a nicotine transporter in leaf vacuoles of Nicotiana tabacum. PMID- 19816141 TI - Integrated analysis of co-expressed MAP kinase substrates in Arabidopsis thaliana. PMID- 19816143 TI - A molecular basis for the physiological variation in shade avoidance responses: a tale of two ecotypes. PMID- 19816144 TI - Unraveling the roles of sphingolipids in plant innate immunity. PMID- 19816145 TI - Chloroplasts autophagy during senescence of individually darkened leaves. PMID- 19816146 TI - Phragmites australis root secreted phytotoxin undergoes photo-degradation to execute severe phytotoxicity. AB - Our study organism, Phragmites australis (common reed), is a unique invader in that both native and introduced lineages are found coexisting in North America. This allows one to make direct assessments of physiological differences between these different subspecies and examine how this relates to invasiveness. Recent efforts to understand plant invasive behavior show that some invasive plants secrete a phytotoxin to ward-off encroachment by neighboring plants (allelopathy) and thus provide the invaders with a competitive edge in a given habitat. Here we show that a varying climatic factor like ultraviolet (UV) light leads to photo degradation of secreted phytotoxin (gallic acid) in P. australis rhizosphere inducing higher mortality of susceptible seedlings. The photo-degraded product of gallic acid (hereafter GA), identified as mesoxalic acid (hereafter MOA), triggered a similar cell death cascade in susceptible seedlings as observed previously with GA. Further, we detected the biological concentrations of MOA in the natural stands of exotic and native P. australis. Our studies also show that the UV degradation of GA is facilitated at an alkaline pH, suggesting that the natural habitat of P. australis may facilitate the photo-degradation of GA. The study highlights the persistence of the photo-degraded phytotoxin in the P. australis's rhizosphere and its inhibitory effects against the native plants. PMID- 19816147 TI - Implication of chlorophyll biosynthesis on chloroplast-to-nucleus retrograde signaling. PMID- 19816148 TI - Tomato Aux/IAA3 and HOOKLESS are important actors of the interplay between auxin and ethylene during apical hook formation. PMID- 19816149 TI - Advances in personalized therapeutics in non-small cell lung cancer: 4q12 amplification, PDGFRA oncogene addiction and sunitinib sensitivity. PMID- 19816150 TI - Nodal diffuse large B-cell lymphomas in children and adolescents: immunohistochemical expression patterns and c-MYC translocation in relation to clinical outcome. AB - Diffuse large B-cell lymphoma (DLBCL) is a very infrequent neoplasm in the pediatric age group; therefore there are very few studies on the immunophenotype or genetics of these cases. We studied a series of 16 patients with nodal DLBCL occurring in patients between 10 and 18 years of age. The cases were classified according to the 2008 World Health Organization classification criteria, with application of immunohistochemistry for the detection of CD10, BCL-6, and MUM1 proteins to divide the lymphomas into germinal center and nongerminal center types. In addition, TCL1, BCL-2 expression, and the Ki-67 proliferation index were evaluated by immunohistochemistry, and c-MYC and BCL2 translocations were evaluated by fluorescence in situ hybridization. All these parameters were correlated with clinical features and outcome. Our study revealed that centroblastic morphology and the germinal center type of DLBCL are more prevalent in these young patients (63%), with 37% containing a c-MYC translocation. Only 1 case showed a BCL2 translocation, reflecting a double-hit case with features intermediate between DLBCL and Burkitt lymphoma. We found a higher frequency of BCL-2 expression than previously reported, with no direct influence on the outcome of the disease in univariate or multivariate analysis. The expression of TCL1 has not been specifically studied in nodal pediatric DLBCL before; we found a 31% incidence of TCL1 expression. MUM1 expression was observed in 44% of the cases and these positive cases showed a significant negative impact on clinical outcome. TCL1 is directly and significantly associated with the presence of c-MYC and a high proliferative index. The germinal center and nongerminal center subtypes showed significant differences for both overall survival and disease free interval. c-MYC translocation was found in 37% of patients, and had a favorable impact on clinical outcome. We conclude that nodal pediatric and adolescent DLBCL are mainly of the germinal center type, with a generally good outcome despite the frequent expression of BCL-2 and the presence of c-MYC translocation. TCL1 expression seems to be associated with a good clinical outcome, whereas MUM1 expression predicts a poor clinical outcome. PMID- 19816151 TI - An operative approach to the treatment of refractory cutaneous nasal sarcoid: a case report and review of the literature. AB - The treatment of cutaneous sarcoid is often frustrating, because lesions may be refractory to treatment, or recurrent. There is little information in the literature regarding the surgical treatment of cutaneous sarcoid. Such treatment may become necessary in cases that do not respond to systemic treatments and create functional impairments. We present a 7-year follow-up of a patient with cutaneous nasal sarcoid that was successfully treated surgically. PMID- 19816152 TI - Paramuscular and paraneural perforators in DIEAP flaps: radiographic findings and clinical application. AB - The computed tomography microangiography revolutionized the planning of abdominal flaps, and enabled us to identify perforators from the deep inferior epigastric system with a medial extramuscular or minimal medial intramuscular trajectory. We define these perforators as paramuscular and paraneural, since their main course is retromuscular and they emerge medial to the medial border of rectus muscle or medial to its motor nerve supply, respectively. Studying the different perforator distributions in the abdomen of 58 patients who underwent breast reconstruction with deep inferior epigastric artery perforator flap, we have recorded that 46.4% of the abdominal walls have a dominant paramuscular/paraneural perforator (25.8% paramuscular and 20.6% paraneural) with an average diameter 1.56 +/- 0.2 mm. Although, the comparison of the rectus muscle morbidity following deep inferior epigastric artery perforator flap dissection based on paramuscular/paraneural and based on other perforator patterns, did not reveal significant differences, paraneural/paramuscular type perforators are not related anatomically to the motor nerves, and the donor site morbidity should be negligible in any operator's hands. PMID- 19816153 TI - Preoperative shock wave therapy reduces ischemic necrosis in an epigastric skin flap model. AB - Extracorporeal shock wave therapy (ESWT) has recently been demonstrated to improve skin flap survival. In all these studies EWST was applied immediately after the surgical intervention. Thus, the purpose of this study was to determine the preoperative effect of ESWT as a noninvasive technique to precondition flap tissue in a rat epigastric skin flap model. EWST and control groups each contained 10 animals. ESWT was applied 7 days before the surgical intervention, whereas the control group received no treatment. Follow-up evaluation was performed on postoperative day 5. The mean area of flap necrosis, expressed as a percentage of the total flap area, was calculated. A significant reduction of the average flap necrosis area was observed in the ESWT group (27.2% +/- 9.6%) compared with the control group (46.1% +/- 7.9% (P < 0.05). In summary, this study indicates that preoperative ESWT may enhance skin flap survival in a rodent model. PMID- 19816154 TI - Transverse Singapore flap for reconstruction of a congenital rectovaginal fistula in an 18-month-old infant. AB - Many different types of flaps have been used for reconstruction of rectovaginal fistulae. This is the first report of using a transverse Singapore flap for repairing a large 3 cm by 3-cm complex congenital rectovaginal fistula in an 18 month-old infant with VATER syndrome. A right transverse Singapore flap was used to reconstruct both the posterior wall of the vagina and the anterior wall of the rectum while simultaneously joining the 2 structures to the perineum. Despite minor postoperative delayed healing which resolved, the patient has maintained a functionally and cosmetically satisfactory result at 20-month follow-up. For complex rectovaginal fistulae reconstruction in the pediatric patient, the transverse Singapore flap is a reliable and viable option for the reconstructive surgeon. PMID- 19816155 TI - Use of autologous micromucosa graft for vaginoplasty in vaginal agenesis. AB - Despite reports on many different techniques, the ideal method of vaginoplasty has not been firmly established. The purpose of our study was to describe the procedure and outcome of creating neovagina lined with autologous micromucosa. Nine patients with vaginal agenesis were reconstructed by our new technique. The mucosal grafts that harvested from the bilateral cheek were minced into 1 mm2 in size by machine and were transplanted to the cavity, which is dissected between the bladder and the rectum. In all cases, a neovagina was well formed, 8 cm in depth (range, 6-10), and 2 fingers in width. The lining was pink-colored, smooth, and was confirmed as mucosal histologically. The perineal area and donor sites were not disturbed postoperation. Of the 9 patients, 4 were married and sexually active. Three of them were satisfied with their sexual life, whereas the remaining one complained the inadequate depth of the neovagina. We conclude that the use of autologous micromucosa to construct a neovagina appears an easy, effective, and minimally invasive method. It deserves further application. PMID- 19816156 TI - Reconstruction of orbital floor fractures: comparison of individual prefabricated titanium implants and calvarial bone grafts. AB - We assessed fresh (< or = 2 weeks) and old (>2 weeks) orbital floor fractures and examined how selection of the implant affected the development of enophthalmos or the treatment of pre-existing enophthalmos. We conducted a retrospective review of 61 cases treated with calvarial bone grafting or individually prefabricated titanium mesh implants. The features of orbital floor fractures and orbital volume (OV) changes were analyzed by a 3-dimensional medical surface rendering image software system. The difference in OV before and after surgery was significant (P < 0.05) in fresh and old fractures treated with either calvarial bone or individually designed titanium mesh. The difference in OV between unaffected and postoperatively affected sides was not significant in the group that received the individually designed titanium mesh, but was significant in the group that received calvarial bone. Spearman rank correlation analysis indicated positive correlations between enophthalmos and postoperative OV changes after implantation of cranial bone in fresh fractures or titanium mesh in fresh and old fractures. The long-term degree of enophthalmos can be predicted with 3 dimensional medical surface rendering. Individual digitally designed titanium mesh is the proper choice of implant material to recover precise OV in fresh or older orbital fractures. PMID- 19816157 TI - Adherence characteristics after abnormal screening results between mammogram and Papanicolaou test groups. AB - The purpose of this study was to evaluate whether the adherence concerns related to follow-up were different between groups of women after abnormal screening results of mammography (Mam) and Papanicolaou (Pap) test. Taiwanese women with abnormal Mam screening results (n = 42) and abnormal Pap test results (n = 109) provided data via a self-administered survey. Data were collected in 2005. After providing informed consent, women completed the AFAR, an 18 item questionnaire that measured the Adherence concerns of Follow-up after Abnormal screening Results. The results showed that (1) for both Mam and Pap test groups, the most important adherence concern for clinical follow-up after abnormal screening results is disease related, followed by others' response and resource accessibility; and (2) in these 2 clinical follow-up groups, women in the Mam group were more likely to be concerned with all of the 18 adherence factors than those in the Pap test group (OR, 2.46-9.86). The most important adherence concern on clinical follow-up after abnormal screening results for Mam and Pap test women is disease related. Mammography women considered significantly more adherence concerns than Pap test women. These differences imply that improving follow-up after Mam or Pap screen may require different interventions. PMID- 19816158 TI - Searching: the lived experience of women with cancer of the vulva. AB - Although the physical adverse effects following surgery for vulval cancer may readily be described, little is known about the psychological, emotional, and social impact. A review of the literature revealed a paucity of studies, as this condition is rare. To explore the lived experiences of women in the United Kingdom with vulval cancer, a qualitative research study was undertaken using the research methodology of interpretive phenomenology, based on the work of Heidegger (The Essence of Human Freedom. London: Continuum; 2005) and van Manen (Researching Lived Experience. Human Science for an Action Sensitive Pedagogy. New York: State University of New York Press; 1990). Thirteen women younger than 50 years were interviewed between 6 months and 5 years after their surgery, and the data were analyzed using framework analysis (Qualitative data analysis for applied policy research. In: Analysing Qualitative Data. London: Routledge; 1994:173-194). This article describes how the concept of searching emerged from the data. It also describes the manner in which the women searched to control their symptoms by their own efforts and by undergoing surgery. The search to understand the truth about their condition and their access to information in the search for meaning and the time element in which this was undertaken was also raised. An underlying consideration is the need for health professionals to help provide the information and support needed to manage this condition. PMID- 19816159 TI - Factors associated with prostate cancer patients' and their spouses' satisfaction with a family-based intervention. AB - Only a few programs are designed to help couples cope with the effects of prostate cancer, and typically, only their intervention outcomes are reported. The purpose of this study was to assess prostate cancer patients' and their spouses' satisfaction with an efficacious supportive-educative, family-based intervention, and factors associated with their satisfaction. We assessed the relationship of overall satisfaction with the intervention to (1) the patients' and spouses' appraisal and the resource and quality-of-life baseline scores and (2) changes in those scores after completing the intervention. Results showed that participants were very satisfied with the program. Patients who had higher scores on baseline measures, indicating more positive appraisal of their illness, better use of resources (eg, coping, self-efficacy), and higher overall quality of life, reported more satisfaction with the intervention. For spouses, few baseline measures were related to their satisfaction; however, spouses who reported positive changes after intervention (less negative appraisal and uncertainty, better communication) reported higher satisfaction with the program. Although satisfied with the program, factors associated with patients' and spouses' satisfaction differed. To translate effective interventions to clinical practice settings, it is important to assess participants' satisfaction with program content and delivery, as well as program outcomes. PMID- 19816160 TI - A pilot study of an automated voice response system and nursing intervention to monitor adherence to oral chemotherapy agents. AB - This study was designed to develop and test a system to monitor adherence with nonhormonal oral chemotherapeutic agents using an automated voice response (AVR) system plus nursing intervention. Participants were patients diagnosed with solid tumor cancers, primarily breast, colon, and lung cancers, who received the Symptom Management Toolkit and participated in an interview for symptom severity, satisfaction, and beliefs about oral agents. Patients received weekly AVR calls, which assessed adherence to oral agents and severity of 15 symptoms. Patients who reported adherence of below 100% of the prescribed oral agents or symptoms of 4 or greater (0-10 scale) for 3 consecutive weeks were called by a nurse for assistance with symptom management and adherence to oral chemotherapy medications. After the 8 weekly AVR calls, patients participated in a follow-up interview and medical record review. Participants were 30 oncology patients who were ambulatory and treated at 2 cancer centers in Midwest United States. The results indicate 23.3% nonadherence rate to oral chemotherapy medications due to symptoms and forgetting to take the medication. An association between symptom management and adherence was found. Symptom severity and beliefs about medications were not significantly different between adherent and nonadherent patients. This pilot study demonstrated the ability to accrue patients for a longitudinal trial and informed intervention design while providing guidance for future interventions and research studies. PMID- 19816161 TI - Support needs of patients with oral cancer and burden to their family caregivers. AB - Oral cancer patients' supportive care needs during the postoperative period can impact their family caregivers' burden. The purposes of this study were to (1) examine patients' perceived levels of supportive care needs, (2) examine caregivers' perceived levels of caregiving burden, and (3) examine the predictive factors for caregivers' caregiving burden with newly diagnosed oral cancer patients during the postoperative period. A cross-sectional, correlational study was conducted. One hundred twenty-two pairs of eligible patients and caregivers were recruited from the otolaryngology and plastic/reconstructive inpatient wards in a medical center in northern Taiwan. A set of questionnaires was used to assess patients' needs, including the Karnofsky's Performance Status Index, Cancer Needs Questionnaire Short Form, Head and Neck Specific Needs Questionnaire, and Background Information Form; the caregivers' burden was evaluated using the Caregiver Reaction Assessment, Inventory of Socially Supportive Behavior, and Characteristics of Care Experiences Form. In general, patients reported having high overall supportive care needs with the highest level in the "health system and information" domain among 6 needs domains. Caregivers had moderate levels of caregiving burden, with the highest level in the "self-esteem" domain among 5 caregiving experience domains. Factors significantly related to those burdens across 5 domains of caregiving burden were caregivers' social support, patients' physical and daily living needs, patients' health system and information needs, and patients' psychological needs. Postoperative oral cancer patients experienced relatively high unmet supportive care needs, and caregivers perceived moderate levels of caregiving burden. Healthcare professionals should systematically assess patients' and caregivers' problems and provide timely supportive care clinically. PMID- 19816162 TI - Differences in symptom clusters identified using occurrence rates versus symptom severity ratings in patients at the end of radiation therapy. AB - The purposes of this study were to identify the number and types of symptom clusters using yes/no responses from the Memorial Symptom Assessment Scale, identify the number and types of symptom clusters using severity scores from the Memorial Symptom Assessment Scale, compare the identified symptom clusters derived using severity scores to those derived using occurrence ratings, and evaluate for differences in symptom cluster severity scores between patients with breast and prostate cancer at the end of radiation therapy. Separate exploratory factor analyses were performed to determine the number of symptom clusters based on symptom occurrence rates and symptom severity ratings. Although specific symptoms within each symptom cluster were not identical, 3 very similar symptom clusters (ie, "mood-cognitive" symptom cluster, "sickness-behavior" symptom cluster, "treatment-related" symptom cluster) were identified regardless of whether occurrence rates or severity ratings were used to create the symptom clusters at the end of radiation therapy. However, the factor solution derived using the severity ratings fit the data better. Significant differences in severity scores for all 3 symptom clusters were found between patients with breast and prostate cancer. For all 3 symptom clusters, the patients with breast cancer had higher symptom cluster severity scores than the patients with prostate cancer. PMID- 19816163 TI - Exploring patient experiences of cancer services in regional Australia. AB - Numerous previous studies have suggested that cancer care in rural and regional Australia is fragmented and that rural and regional patients have less access to services. However, knowledge concerning inequality of health and cancer care in regional and rural settings is still inconclusive. The social and economic disadvantages in regional locations are often linked to insufficient resources and infrastructures to support cancer care and treatment. The aim of this study was to explore cancer patients' experiences from the perspective of patients and healthcare providers in a regional area of Western Australia. It was found that patients who had the involvement of a cancer nurse coordinator and cancer support workers had better experiences and more streamlined care than did those who had to navigate the journey alone. This study endorses the function of cancer nurse coordinators and cancer support workers in providing better coordination of care in rural and regional cancer patients. PMID- 19816164 TI - Construction of palliative care training contents in China: a Delphi study. AB - There is a growing emphasis on improving the quality of ending life in China today; however, Chinese professional nursing education in palliative care is limited and inconsistent, and there are no standardized formal training contents. There is a need to construct a related training mode to help Chinese nurses to improve their quality of palliative care. The objective of this study was to construct palliative-care-related training contents for Chinese clinical nurses. A total of 93 training items related to palliative care were developed by literature review. A 36-member expert panel engaged in a 2-round Delphi process to assess the importance of these 93 items and to identify any additional important elements. The expert panelists reached consensus on 69 training items that belonged to 6 modules; these were (1) 8 items for the palliative care overview, (2) 24 items for symptom care, (3) 15 items for psychological care, (4) 13 items for communication and exchange, (5) 5 items for ethics and laws, and (6) 4 items for terminal care. This study formed a 6-module training content through Delphi study, which will provide the basic guideline for Chinese nurses to establish their core competence about palliative care. The 6-module integrated training mode achieved in this study represents the core knowledge and skill that a nurse who provides palliative care in China needs to know; then the nurse can choose any module to get training according to his or her practical needs and availability, totally or separately. PMID- 19816165 TI - The validity and reliability of Champion's Health Belief Model Scale for breast cancer screening behaviors among Iranian women. AB - Breast cancer is a global issue that continues to be the most diagnosed cancer in women. The incidence of breast cancer is increasing, and the incidence age for Iranian women is at least 10 years earlier than in western countries. Beliefs related to breast cancer have been found to be a factor in a woman's decision about breast screening behavior, and a valid and reliable questionnaire is necessary to the development of education interventions in this area. The aim of the current study was to translate the Champion Health Belief Model Scale to Farsi and to examine the psychometric properties of the Farsi version. A random sample of 606 employed women (20-69 years old) from Sanandaj, Iran, participated in the study. Construct validity of the Farsi version was supported through factor analysis. Nine factors emerged for breast self-examination (2 barriers factors, 2 benefits factors, 2 confidence factors, and 1 factor each related to seriousness, motivation, and susceptibly) and 6 factors related to mammography (barriers, seriousness, susceptibly, benefits, and 2 motivation factors). All items loaded on their respective factors except 1 item. It was concluded that the Farsi version of the Champion Health Belief Model Scale has the potential to measure beliefs related to breast self-examination and mammography with Iranian women. Further evaluation of the measure with different populations is warranted. PMID- 19816166 TI - Health-related quality of life in adults with Hodgkin's disease: the state of the science. AB - Hodgkin's disease (HD) affects younger and older adults and can disrupt developmental tasks and cause multiple medical sequelae. Since long-term survival is excellent, understanding issues related to all domains of health-related quality of life (HRQOL)-physical, psychological, social/functional, and spiritual after completion of treatment is a critical step in designing and testing interventions to improve survivors' adjustment and return to their previous level of functioning. This article is an integrative review of empirical studies of HRQOL in HD survivors. Following Ganong's guidelines, 35 studies were identified and reviewed. Commonly reported physical consequences of HD include fatigue, anticipatory nausea and vomiting, and cognitive problems that lasted several years after treatment completion, as well as long-term life-threatening adverse effects including secondary cancers and cardiovascular and respiratory complications. Psychological consequences include emotional distress, especially depression and anxiety, and social/functional difficulty, including inability to return to work and adjustment to the workplace environment secondary to diminished capacity to complete work tasks. Within the spiritual domain, survivors reported that they had a greater appreciation for life after treatment. Development of appropriate theory-guided interventions to improve the HRQOL for HD survivors can be achieved through more rigorous study designs and standardization of HRQOL measurements. PMID- 19816167 TI - Initial report of a family registry of multiple myeloma. AB - About 20,000 Americans are diagnosed with multiple myeloma (MM) each year, and more than 10,000 die of MM in the United States annually. The etiology of MM remains unknown, although genetic and environmental factors have been implicated. Patients (n = 68) from the Myeloma Institute for Research and Therapy at the University of Arkansas for Medical Sciences and their family members with MM or a related malignancy were interviewed for environmental factors associated with MM and for family history data to complete pedigrees. In collaboration with Dr Henry Lynch at Creighton University, pedigrees of at least 3 generations were analyzed. Eighteen families (27%) have a putative autosomal dominant mode of genetic transmission of MM. Furthermore, the pedigrees indicate that pancreatic cancer, malignant melanoma, breast cancer, and lymphoma may be part of a myeloma syndrome. Environmental factors associated with MM present in this patient population were being born and raised in a rural area, raising cattle or cotton, and exposure to pesticides, insecticides, or herbicides. This work will be part of the efforts to create an international consortium to study familial MM. Research in the area of molecular epidemiology is needed to discover the genetic and environmental determinants of this disease. PMID- 19816168 TI - Management of inherited thrombophilia in pregnancy. AB - PURPOSE OF REVIEW: This review summarizes the currently available data concerning risk and management of venous thromboembolism in pregnant women with inherited thrombophilia. SUMMARY: Pregnancy is a hypercoagulable state, and inherited thrombophilia increases this risk further. Despite the risks, the actual incidence of venous thromboembolism remains low, and therefore, the widespread use of anticoagulants for pregnant women with inherited thrombophilia is not advised. Although randomized, placebo-controlled trials investigating the risks and benefits of anticoagulation have not been performed, there are data to support the use of low molecular weight heparin for high and intermediate-risk women. We will review these data and treatment recommendations, which are based on retrospective and case-control studies as well as expert opinion and consensus statements. PMID- 19816169 TI - 'Old' bones in young bodies: the tale of cystic fibrosis. AB - PURPOSE OF REVIEW: Cystic fibrosis (CF) is the most common genetic disease within the white population and leads to premature respiratory failure. Approximately, 60 000 individuals are currently living with CF in North America and Europe, almost half of whom are adults. RECENT FINDINGS: Dozens of studies across the globe indicate that CF adults have low bone density and increased rates of fractures. This genesis of the problem appears to be in late childhood to adolescence. SUMMARY: Prevention and treatment of CF-related bone disease must address the myriad risk factors (decreased absorption of fat-soluble vitamins due to pancreatic insufficiency, altered sex hormone production, chronic lung infection with increased serum levels of proinflammatory, bone-active cytokines, malnutrition and low body weight, physical inactivity and glucocorticoid therapy) for poor bone health. This review will address the pathogenesis, diagnosis and treatment of bone disease in CF. It will also discuss best practice guidelines for optimizing bone health in patients with CF. PMID- 19816170 TI - Role of the pituitary-bone axis in skeletal pathophysiology. AB - PURPOSE OF REVIEW: Embedded within textbooks for decades is the hard fact that releasing hormones from the anterior pituitary, namely, follicle-stimulating hormone, thyroid-stimulating hormone and adrenocorticotropic hormone, stimulate master hormone secretion from target endocrine organs. We propose a paradigm shift in endocrine physiology, which is that these hormones act by design on bone directly, also now considered an endocrine organ. RECENT FINDINGS: Complementary investigations using mouse genetic and cell biological approaches reveal that follicle-stimulating hormone and thyroid-stimulating hormone act on bone cells directly to regulate bone remodeling and bone mass. Thyroid-stimulating hormone inhibits bone remodeling, whereas follicle-stimulating hormone stimulates it. We also find that the posterior pituitary hormone oxytocin is anabolic to the skeleton. SUMMARY: An ambitious extrapolation is that a plurality of pituitary hormones acts in concert as part of a 'pituitary-bone' axis to regulate skeletal integrity in health and disease. When dysregulated master hormone levels during hypogonadism and hyperthyroidism cause altered pituitary hormone secretion through hypothalamic feedback, the latter hormones contribute to the skeletal loss. PMID- 19816171 TI - Neuropeptides and inflammatory bowel disease. AB - PURPOSE OF REVIEW: Inflammatory bowel disease (IBD) is a chronic intestinal inflammatory condition, the pathophysiology of which is not well understood. It has, however, become increasingly evident that interactions between the enteric nervous system and the immune system play an important role in the cause of IBD. Both the enteric nervous system and the central nervous system can amplify or modulate the aspects of intestinal inflammation through secretion of neuropeptides or small molecules. The purpose of this study is to present recent data on the role that neuropeptides play in the pathophysiology of IBD. RECENT FINDINGS: The best studied of the neuropeptides thought to play a role in the pathogenesis of IBD include substance P, corticotropin-releasing hormone, neurotensin, and vasoactive intestinal peptide; small molecules include acetylcholine and serotonin. Recently discovered functions of each of these neuropeptides with a discussion of implications of the data for therapy are reviewed. SUMMARY: Although the available data suggest an important role for neuropeptides in the pathophysiology of intestinal inflammation, there does yet not appear to be a function that can be taken as established for any of these molecules. The complexity of neuroimmune-endocrine systems, conflicting study results and dual mechanisms of action, warrant further research in this field. Clarification of the molecular mechanisms of action of neuropeptides and on immune and inflammatory reactions will likely yield new treatment options in the future. PMID- 19816172 TI - Gastric oncology: an update. AB - PURPOSE OF REVIEW: As the field of endoscopy progresses, new tools and techniques continue to be developed for gastroenterology in general and gastrointestinal oncology in particular. Some of these include enhancements in diagnostic optics such as chromoendoscopy, magnification endoscopy, and confocal laser endomicroscopy. Others include advanced therapeutics such as endoscopic mucosal resection and endoscopic submucosal dissection. In this review, we will update the reader on these latest of technologies, their benefits and risks, as well as their role in evaluating, staging, and treating gastric neoplasms, especially gastric adenocarcinoma, gastrointestinal stromal tumors, and primary gastric lymphoma. RECENT FINDINGS: Noteworthy studies in this review indicate that in properly selected patients with gastric adenocarcinoma, endoscopic submucosal dissection is a viable alternative to gastric resection with 100% 5-year survival rates; in patients with metastatic gastrointestinal stromal tumors, imatinib can provide effective treatment with reasonable outcome; and in patients with low grade mucosa-associated lymphoid tissue lymphoma, eradication therapy with antibiotics is curative with a very low recurrence rate. SUMMARY: The advances discussed in this review have significantly improved the care we can offer our patients in gastric oncology. With continued advancement in the field, it will be crucial to continue to study outcomes and safety of these techniques and to develop structured training for those looking to perform these procedures. PMID- 19816173 TI - Vasculotropic effects of calcimimetics. AB - PURPOSE OF REVIEW: At all stages of chronic kidney disease (CKD) cardiovascular death is the most prominent cause of mortality. Current treatment options are still not completely satisfactory in this group of high cardiovascular risk patients. Experimental data and clinical observations suggest a role of secondary hyperparathyroidism, hyperphosphatemia, and hypercalcemia in the genesis of cardiovascular complications of CKD. The ubiquitous expression of the calcium sensing receptor, which is targeted by calcimimetics and the pleiotropic effects of calcimimetics, make this class of drugs potential candidates for cardiovascular intervention. RECENT FINDINGS: Recent experimental studies suggest that calcimimetics interfere with the development of vascular abnormalities in CKD and to some extent even reverse them. The effects of calcimimetics on the vasculature are, at least partially, independent of their effects on calcemia, phosphatemia, and parathyroid hormone concentration. The beneficial effects of calcimimetics on vascular calcification, arteriolar thickening, atherogenesis, and myocardial capillarization are well documented. In addition they have hypotensive and renoprotective actions. SUMMARY: Experimental models suggest beneficial effects of calcimimetics on cardiovascular disease. Although prospective clinical data are still lacking, retrospective data suggest cardiovascular benefit of calcimimetics even in humans. Clinical trials with calcimimetics evaluating hard cardiovascular end-points would be desirable. PMID- 19816174 TI - Acid-base regulation of angiotensin receptors in the kidney. AB - PURPOSE OF REVIEW: Angiotensin II (Ang II) is a major systemic regulator of blood pressure, blood volume, and acid-base homeostasis. However, the effects of local Ang II signaling in the kidney are less well known. Here we review the impact of Ang II on acid-base transport in specific nephron segments and discuss the recently described role of acid loading in sensitizing the proximal tubule to Ang II. RECENT FINDINGS: Recent studies suggest that Ang II signaling is necessary for a normal kidney response to acid loads. Under acidic conditions, tubule cells upregulate type 1 Ang II receptors, amplifying the effects of Ang II. Site of action may also affect Ang II activity, as Ang II can produce divergent effects when acting on the luminal versus basolateral aspects of tubule cells. High luminal concentrations of Ang II and its precursors may facilitate coordinated acid-base transport and metabolic activity throughout the nephron. SUMMARY: Local Ang II signaling in the kidney directly modulates acid-base transport and metabolism, providing an additional mechanism by which the renin-angiotensin aldosterone system regulates acid-base balance. The segment-specific and acid stimulated responses of the tubule to Ang II are diverse and merit further exploration for their adaptive and potentially maladaptive roles. PMID- 19816175 TI - Cataract surgery in pseudoexfoliation syndrome. AB - PURPOSE OF REVIEW: To discuss the preoperative, intraoperative and postoperative considerations and techniques for cataract surgery in pseudoexfoliation syndrome. RECENT FINDINGS: Pseudoexfoliation syndrome is a common disorder worldwide that has been linked to the lysyl oxidase-like-one (LOXL1) gene. Its identification in patients undergoing cataract surgery has become important in preventing complications through proper preoperative planning. The management of the small pupil found in pseudoexfoliation can be achieved using various pharmacological and mechanical techniques. There are now several devices available to support the weak zonules that commonly accompany pseudoexfoliation. Postoperatively, pseudoexfoliation patients need to be followed closely to monitor for complications such as intraocular pressure spikes, inflammation and intraocular lens dislocation. SUMMARY: With proper preoperative preparation, intraoperative use of appropriate adjunctive devices, and close postoperative follow-up, favorable outcomes can be achieved in cataract surgery in pseudoexfoliation syndrome. PMID- 19816176 TI - Visual aspects of neurologic protein misfolding disorders. AB - PURPOSE OF REVIEW: This article reviews topics of interest to the ophthalmologist relating to the most common neurologic protein misfolding disorders. RECENT FINDINGS: Many neurodegenerative diseases are pathologically associated with misfolded proteins. These diseases cause a profound impact of disability to the individual and society. Alzheimer's disease costs alone are estimated to be over US$225 billion annually in the USA. The ophthalmologist is often asked to provide an opinion regarding the cause of visual symptoms in patients with these unique disorders. SUMMARY: The categorization of neurodegenerative diseases has evolved based on advances in genetic, molecular and pathological research. In many neurodegenerative diseases, aggregation of a misfolded protein is responsible for the development of pathologic inclusions. When the misfolded protein is tau or synuclein, these diseases are called tauopathies or synucleinopathies, respectively. This article focuses on ophthalmic findings in some of the most common tauopathies and synucleinopathies: Alzheimer's disease, progressive supranuclear palsy, Parkinson's disease, dementia with Lewy bodies and multisystem atrophy. PMID- 19816177 TI - Multiple sclerosis. Part 2: ophthalmic issues in MS therapy. AB - PURPOSE OF REVIEW: This update includes topics relating to the treatment of multiple sclerosis (MS) for the ophthalmologist. RECENT FINDINGS: Despite investigation into numerous genetic and environmental factors, the cause of MS remains elusive. MS is a disease of the central nervous system that mainly affects young people and usually progresses over time, leading to disability in multiple areas, including mobility, cognition, affect and vision. It is likely that, as novel MS treatments are introduced, there will be a greater interest in tracking visual function and monitoring for visual complications. SUMMARY: Current therapies in MS include interferon, glatiramer, natalizumab and mitoxantrone. Other MS treatments are imminent, and include oral medications, monoclonal antibodies and chemotherapy. Treatment of the visual symptoms in MS and the side effects of MS therapies are reviewed in this article. PMID- 19816178 TI - Viral-induced wheezing episodes in preschool children: approaches to therapy. AB - PURPOSE OF REVIEW: The high prevalence of wheezing illnesses in the preschool age group and the heterogeneity of wheezing phenotypes early in life combine to present significant challenges in the clinical management of this problem. RECENT FINDINGS: Recent studies have examined multiple therapeutic strategies to reduce the frequency and/or severity of viral-triggered wheezing episodes in young children. Clinical trials demonstrate various degrees of clinical benefit associated with daily use of inhaled corticosteroids or leukotriene receptor antagonists in terms of episode prevention and attenuation, and with episodic therapy with high-dose inhaled corticosteroids or leukotriene receptor antagonists in terms of episode attenuation. Recent evidence has brought into question the efficacy of systemic corticosteroid therapy as an element of inpatient care for viral-induced wheezing. SUMMARY: The optimal management strategy for virus-induced wheezing in early life remains elusive. Future research in this area should incorporate consideration of the heterogeneous nature of this complex syndrome. PMID- 19816179 TI - Novel bronchodilators in asthma. AB - PURPOSE OF REVIEW: Because of the central role of bronchodilators in the treatment of asthma, in recent years there has been a renewed interest in the field and now once-daily bronchodilators are in development in an attempt to simplify their use. RECENT FINDINGS: A variety of beta2-agonists with long half lives, also called ultra long-acting beta2-agonists (ultra-LABAs; indacaterol, carmoterol, milveterol, GSK-642444, BI-1744-CL, LAS-100977, and PF-00610355) are currently under development with the hopes of achieving once-daily dosing. Between them, indacaterol, GSK-642444, and carmoterol are at a more advanced stage of development. Also several novel inhaled long-acting antimuscarinic agents are currently being developed, but their role in the treatment of asthma is limited. As combination therapy with an inhaled corticosteroid (ICS) and a LABA is considered the first-line approach for treating patients suffering from asthma, some novel once-daily combinations of LABAs and ICSs are under development. SUMMARY: Bronchodilators are central in the symptomatic management of asthma. It is likely that the once-daily dosing of a bronchodilator would be a significant convenience and probably a compliance-enhancing advantage, leading to improved overall clinical outcomes in patients with asthma. In any case, as a LABA in combination with an ICS continues to be the most effective asthma treatment, once-daily combinations of ultra-LABAs and ICSs will be central in the treatment of asthmatic patients in the next years. PMID- 19816180 TI - Highly variable population-based prevalence rates of unilateral hearing loss after the application of common case definitions. AB - OBJECTIVES: This study shows how population-based estimates of the prevalence of unilateral hearing loss (UHL) in children aged 6 to 19 yrs can differ considerably with various applications of commonly accepted case definitions. It also examines demographic variables and risk factors related to UHL. DESIGN: The Third National Health and Nutrition Examination Survey, conducted from 1988 to 1994, is a national population-based, cross-sectional survey. This study examined results of audiometric testing at 0.5 to 8 kHz and demographic data from in person examination interviews. Three definitions of UHL were used: (1) 0.5, 1, and 2 kHz > or = 15 dB pure-tone average (PTA); (2) 0.5, 1, 2, and 4 kHz > or = 15 dB PTA; and (3) 0.5, 1, and 2 kHz > or = 20 dB or PTA >25 dB at two or more frequencies above 2 kHz (3, 4, 6, and 8 kHz). Case definitions 2 and 3 are not merely subsets of case definition 1. Some overlap exists between the groups, but each case definition classifies a proportion of children who fall uniquely under that case definition. Inclusion of participants based on tympanometry results (test of middle ear function) was also examined as were demographic characteristics and risk factors associated with UHL. RESULTS: Overall, the weighted proportion of children with UHL using case definition 1 was 6.3% (approximately 3,213,000 children nationally); using case definition 2, it was 5.8% (approximately 2,958,000 nationally); using case definition 3, it was 3.0% (approximately 1,530,000 nationally). For all three case definitions, children who failed tympanometry were at higher risk for UHL than children who passed. For case definition 2, children from rural areas were at higher risk for UHL than were children from urban areas. CONCLUSIONS: This study demonstrates that different applications of well-accepted case definitions of UHL can influence population-based prevalence estimates, in this study by as much as a factor of 2. These findings highlight the importance of controlling for tympanometry status as a risk factor in such estimates. Which demographic characteristics and risk factors are significantly associated with hearing loss seem to vary depending on the case definition. These findings have implications for the interpretation of prevalence rates and risk factors in the literature on hearing loss in general. Prevalence rate estimates require careful consideration of the case definition of hearing loss, tympanometry status, and demographic characteristics. PMID- 19816181 TI - Temporal resolution in regions of normal hearing and speech perception in noise for adults with sloping high-frequency hearing loss. AB - OBJECTIVES: (1) To determine whether high-frequency sensorineural hearing loss (HF SNHL) is accompanied by deterioration in temporal resolution in the low frequency region where hearing sensitivity is within normal range. (2) To evaluate whether such temporal processing deficits contribute to speech perception difficulty in noise. DESIGN: A between-group design was employed, using subjects either with or without high-frequency hearing loss and matched by age. Temporal resolution was evaluated in amplitude modulation (AM) detection and gap detection tasks. To restrict evaluation to the low-frequency regions where the auditory sensitivity was virtually normal, low-pass noise carriers (for AM detection) and gap markers (for gap detection) were used. The impact of temporal processing deficits on speech perception was evaluated using hearing in noise tests (HINT) with varied time compression rates of the speech materials. RESULTS: Adults with high-frequency hearing loss showed poorer performance than the age matched normal-hearing subjects on both the AM and gap detection tasks, even though the stimuli were restricted to regions of observed normal sensitivity. With increasing time compression, listeners with HF SNHL required a larger signal to noise ratio to maintain accuracy in speech perception in adaptive HINT and exhibited a bigger decrease in score for HINTs at a fixed signal to noise ratio. Multiple regression/correlation analyses show significant correlation across the scores of AM/gap detection tasks and HINTs. CONCLUSIONS: Temporal resolution in the low-frequency region with near-normal sensitivity seems to be deteriorated in subjects with HF SNHL. They were more sensitive to increases in speech rate, suggesting that poorer temporal processing may be related to speech perception deficits in noise. PMID- 19816182 TI - Spectral integration and bandwidth effects on speech recognition in school-aged children and adults. AB - OBJECTIVES: Previous studies have shown that adult listeners are more adept than child listeners at identifying spectrally degraded speech. However, the development of the ability to combine speech information from different frequency regions has received little previous attention. The purpose of this study was to determine the effect of age on the bandwidth necessary to achieve a relatively low-criterion level of speech recognition for two frequency bands and then to determine the improvement in speech recognition that resulted when both speech bands were present simultaneously. DESIGN: Listeners in this study included normal-hearing children (aged 6 to 14 yrs, N = 18) and adults (N = 11). In the first stage of testing, sentences were band pass filtered around either 500 or 2500 Hz, and the bandwidth of that filter was varied adaptively to determine the width required for approximately 15 to 25% correct speech recognition. In the second stage of testing, these criterion bandwidths were presented in fixed block trials with either one band or both bands, and the percent correct performance was determined. RESULTS: Results suggest that age is inversely associated with the bandwidth required to achieve a relatively low-criterion level of speech recognition for speech bands centered at either 500 or 2500 Hz. However, both adults and children show a similar, large improvement in performance when both bands are presented simultaneously. CONCLUSIONS: Although younger children require more bandwidth to recognize speech filtered around a single frequency correctly, they seem to be relatively adept at integrating frequency-distributed information to recognize a composite stimulus. PMID- 19816183 TI - In vitro adsorption of tear proteins to hydroxyethyl methacrylate-based contact lens materials. AB - OBJECTIVES: Investigations of polymer interactions in single protein solutions is a necessary step in the elucidation of in vivo early binding events during protein deposition on hydroxyethyl methacrylate-based contact lens materials. Quantity and tenacity of binding of significant tear components to groups I and IV contact lenses was assessed. Competitive binding by these components was also examined. METHODS: Adsorption on FDA groups I and IV hydrogel lenses was monitored using I-labeled protein. Lenses were incubated in increasing concentrations of radiolabeled single species proteins in solution. For competition experiments, concentration of each radiolabeled protein was held constant and the adsorption/sorption challenged with increasing concentrations of nonlabeled proteins. Lenses were soaked in phosphate-buffered saline to determine desorption. RESULTS: Group IV lenses bound large amounts of lysozyme, whereas group I lenses bound highest amounts of albumin. Albumin binding to both lens types was relatively strong and could not be competed from binding by other proteins lysozyme, lactoferrin, and mucin. Mucin at high concentrations tended to positively cooperate with the binding of lactoferrin and albumin to all lenses. CONCLUSIONS: Binding of proteins to hydroxyethyl methacrylate-based hydrogel lens surfaces is affected by charge and polymer components, and perhaps manufacturing processes. Albumin binds strongly to lens surfaces, and this may play an adverse role during contact lens wear. PMID- 19816184 TI - Scedosporium scleritis or keratitis or both: case series. AB - PURPOSE: To report case series of infectious scleritis or keratitis or both caused by Scedosporium fungal species. METHODS: Medical records of patients presenting with infectious scleritis or keratitis or both caused by Scedosporium were retrospectively reviewed at a tertiary care hospital. Nine culture-proven cases of Scedosporium corneal and scleral infection that were treated medically or surgically or both were enrolled for the study. The main parameters evaluated were the predisposing factors, clinical presentation, and the success of various treatment modalities. RESULTS: The most common mode of presentation was scleritis (six cases, 67%), followed by sclerokeratitis (two cases, 22%) and keratitis (one cases, 11%). The risk factors included a previous pterygium excision with or without beta-radiation (six cases, 67%) and trauma (two cases, 22%). One patient had no identifiable risk factor. Mean duration between the time of pterygium excision and onset of infectious scleritis or keratitis or both was 7.6 +/- 3.3 years (range, 5-12 years). Voriconazole was the most commonly used antifungal medication in the form of drops (five cases, 56%) or tablets (five cases, 56%). Medical therapy alone could successfully resolve infection in six cases. Penetrating keratoplasty was performed in one eye, and two eyes were enucleated. CONCLUSIONS: Corneal and scleral infection caused by Scedosporium may occur many years after pterygium surgery. Although newer antifungal agents like voriconazole are effective in management of such cases, severe infections may result in the loss of eye. PMID- 19816185 TI - Demographics of patients older than 50 years with keratoconus. AB - PURPOSE: This study was designed to determine the demographics of patients aged 50 years or more with a diagnosis of keratoconus, who were seen on the Cornea Service at Wills Eye Hospital. METHODS: A retrospective chart review of all patients with the diagnosis of keratoconus was undertaken to identify demographics characteristics between January 1, 2005 and December 31, 2005. RESULTS: A total of 697 patients were identified. Two hundred seventy-nine (40.0%) patients were age 50 years or more. The age ranged from 50 to 93 years (average 60.2 +/- 8.2 years). Keratoconus was diagnosed as a bilateral disease in 266 (95.3%) and as a unilateral disease in 13 patients (4.7%). In 186 patients (66.6% of the total), the average age at the time of diagnosis was 31.7 +/- 10.9 years (range from 13 to 70 years). Of the 279 patients, 167 had surgery (59.8%). In the group of 112 patients that did not have surgery, 25 (9.0%) were treated with glasses in both eyes, 85 (30.5%) with contact lenses in both eyes, and 2 (0.7%) with glasses in one eye and contact lenses in the other eye. The average time of follow-up was 13.6 +/- 10.5 years (range from 0.1 to 38.7 years). CONCLUSIONS: In a corneal referral practice, the number of patients older than 50 years with keratoconus is higher than previously reported in the literature. We believe that this is because our practice has a strong interest in both contact lenses and corneal surgery with the ability to follow up patients on a long-term basis. PMID- 19816186 TI - Residual corneal flattening after discontinuation of long-term orthokeratology lens wear in asian children. AB - OBJECTIVES: Most patients treated with orthokeratology (OK) in Asia are children. The effects of long-term OK on corneal curvature among children after lens wear discontinuation have not previously been reported. METHODS: This study investigated 28 subjects, aged 10.4 +/- 2.5 years at commencement of OK lens wear, who had been treated with OK for 50.2 +/- 27.4 months. Corneal curvature was measured with a Canon RK3 autokeratometer before commencement of OK and again approximately 2 weeks (17.3 +/- 8.6 days) after discontinuation of OK lens wear. Data were analyzed using paired Student t tests, with a critical P value of 0.05. RESULTS: Compared with baseline, residual flattening of the cornea was found after cessation of OK lens wear, averaging 0.07 +/- 0.04 mm in the flat meridian (P<0.001) and 0.02 +/- 0.07 mm in the steep meridian (P=0.07). This was confirmed by power vector analysis of curvature data, with a significant reduction in M ( 0.27 +/- 0.23 diopter (D); P<0.001), and a slight increase in J0 (0.17D +/- 0.36D; P=0.02). A trend toward greater residual corneal flattening was found among subjects with more pretreatment myopic refractive error (P=0.04). CONCLUSIONS: Corneal curvature may not necessarily return completely to baseline after discontinuation of long-term OK lens wear in children. Intermittent temporary discontinuation of OK lens wear and close monitoring of corneal recovery are recommended during long-term OK treatment. PMID- 19816187 TI - Induced astigmatism after diamond burr superficial keratectomy for recurrent corneal erosion. AB - PURPOSE: To report a case of induced astigmatism after diamond burr superficial keratectomy (DBSK) for recurrent corneal erosion (RCE). DESIGN: Case report. METHODS: Review of clinical findings in a 54-year-old women with ocular history of a scleral buckling procedure for a retinal detachment from blunt trauma and phacoemulsification with intraocular lens placement. The patient presented with RCE after trauma with a mascara brush to the OD and was treated with DBSK. Postoperatively, she developed significant astigmatism. RESULTS: In the third postoperative week after the DBSK procedure, the patient reported of worsening vision. On corneal topography, the patient was found to have 4 diopters of induced astigmatism. The astigmatic error was followed closely by serial corneal topography; a gradual decrease in the amount of astigmatism occurred over the course of 30 weeks. Forme fruste keratoconus was suspected in the patient's contralateral eye, based on corneal topographic analysis. CONCLUSIONS: Induced corneal astigmatism is a previously undescribed complication that can occur after DBSK. It is unclear whether the induced astigmatism in our patient was caused by the DBSK procedure alone or whether the patient had decompensated structural integrity from forme fruste keratoconus or blunt corneal trauma or both. The authors recommend that corneal topographic analysis be appropriately considered before DBSK for RCE and that corneal astigmatism be seen as a potential complication of the procedure. PMID- 19816188 TI - Generation of HCMV-specific T-cell lines from seropositive solid-organ-transplant recipients for adoptive T-cell therapy. AB - Chronically immunosuppressed patients, like solid-organ-transplant (SOT) recipients, are at increased risk for severe human cytomegalovirus (HCMV) infection. Despite the availability of effective antiviral drugs, lasting control of remaining viruses is dependent on an effective T-cell immunity. So in some patients conventional antiviral therapy cannot control the infection and prolonged virostatic therapy is limited by its side effects and the development of viral resistance. Selective reconstitution of cellular immunity by adoptive transfer of HCMV-specific T cells derived from healthy donors is a safe and effective approach in hematopoietic stem cell transplant recipients. The aim of this study was to determine whether functional HCMV-specific T cells can also be generated from chronically immunosuppressed patients. Autologous CD4+/8+ T-cell lines directed against the HCMV protein IE-1 were generated from the peripheral blood of SOT patients using a recently developed modular protocol easily applicable to good-manufacturing-practice conditions. T-cell lines from SOT patients showed similar features as cells from healthy donors regarding phenotype, functionality (HCMV-specific killing, gene expression pattern, and cytokine secretion), IE-1 epitope recognition, and dominance of distinct T-cell receptor V beta families. Most importantly, this protocol also allowed the generation of T-cell lines from immunosuppressed patients with HCMV infection/chronic HCMV disease. Our data suggest the potential of this autologous approach for the treatment of SOT recipients suffering from HCMV infection/disease poorly responding to virostatic therapy. PMID- 19816189 TI - IL-18 paradox in pancreatic carcinoma: elevated serum levels of free IL-18 are correlated with poor survival. AB - The role of the proinflammatory interleukin (IL)-18 in cancer progression remains controversial; we thus examined the hypothesis that impaired antitumor immune response in pancreatic carcinoma patients is related to elevated levels of its natural inhibitor IL-18 binding protein (BP) and/or to alteration in the IL-18 receptor complex expression and function. IL-18 and IL-18 binding protein isoform a (BPa) was assessed in pancreatic carcinoma patients at various disease stages, and after surgery/chemotherapy; free bioactive IL-18 concentrations were calculated. IL-18 receptor complex expression in lymphocyte subsets was analyzed and signaling function was assessed versus healthy donors. Carcinoma cells exhibited below normal IL-18BPa expression and above normal IL-18 expression. Circulating IL-18BPa and IL-18 were above controls. Unexpectedly, free unbound IL 18 serum levels were correlated with disease severity and poor survival. IL-18BPa levels were unchanged by surgery but free IL-18 levels were elevated. Gemcitabine with 5-fluorouracil or oxaliplatin, but not alone, increased IL-18 and free IL-18 levels statistically significantly, without affecting IL-18BPa. Spontaneous/induced IL-18 receptor alpha and receptor beta expression in peripheral blood lymphocyte subsets from patients with advanced disease were near normal, although CD4+ and CD8+ cells were fewer in percentage, and fully functional in inducing interferon-gamma. IL-18 is proposed as novel adjuvant cancer therapy, but free IL-18 levels are increased in the blood of pancreatic carcinoma patients, despite elevated IL-18BP levels, and are associated with poor survival; this highlights recent experimental insights into the prometastatic and proangiogenic effects of IL-18, and suggests that careful preclinical studies are needed to determine the proper application of IL-18 in cancer therapy. PMID- 19816190 TI - Intralesional lymphokine-activated killer cells as adjuvant therapy for primary glioblastoma. AB - Despite recent advances, median survival for patients with resectable glioblastoma multiforme (GBM) is only 12 to 15 months. We previously observed minimal toxicity and a 9.0-month median survival after treatment with intralesional autologous lymphokine-activated killer (LAK) cells in 40 patients with recurrent GBM. In this study, GBM patients were treated with adjuvant intralesional LAK cells. Eligible patients had completed primary therapy for GBM without disease progression. LAK cells were produced by incubating autologous peripheral blood mononuclear cells with interleukin-2 for 3 to 7 days and then placed into the surgically exposed tumor cavity by a neurosurgeon. The 19 men and 14 women had a median age of 57 years. Prior therapy included surgical resection (97%), partial brain irradiation (97%), gamma knife radiosurgery (97%), and temozolomide chemotherapy (70%). Median time from diagnosis to LAK cell therapy was 5.3 months (range: 3.0 to 11.1 mo). LAK cell treatment was well tolerated; average length of hospitalization was 3 days. At the time of this analysis, 27 patients have died; the median survival from the date of original diagnosis is 20.5 months with a 1-year survival rate of 75%. In subset analyses, superior survival was observed for patients who received higher numbers of CD3+/CD16+/CD56+ (T-LAK) cells in the cell products, which was associated with not taking corticosteroids in the month before leukopheresis. Intralesional LAK cell therapy is safe and the survival sufficiently encouraging to warrant further evaluation in a randomized phase 2 trial of intralesional therapies with LAK or carmustine-impregnated wafers. PMID- 19816191 TI - Human bone marrow as a source to generate CMV-specific CD4+ T cells with multifunctional capacity. AB - The bone marrow (BM) is an important compartment for T cell memory. In cytomegalovirus (CMV)-seropositive individuals peripheral blood (PB) CMV-specific T cells constitute a large fraction of PB T cells but are mostly differentiated effector/effector memory T cells with limited survival and proliferative potential. In this study, we performed a comprehensive analysis of the CMV specific T cell response in BM studying both CD4+ and CD8+ T cell responses against overlapping peptide pools of the CMV proteins pp65 and immediate early protein-1. CMV-specific T cell responses were characterized ex vivo and after in vitro expansion of paired PB/BM samples by multiparameter flow cytometry determining surface phenotype, cytokine profile, and cytotoxic capability. Comparable frequencies of CMV-specific T cells were found in un-manipulated PB and BM. Both total CD4+ and CD8+ T cells could be more rapidly expanded from BM. Expanded BM T cells contained significantly higher frequencies of CMV-specific CD4+ T cells than PB. Furthermore, higher frequencies of specific CD4+ T cells from BM were multifunctional, characterized by simultaneous production of interferon-gamma, tumor necrosis factor, and interleukin-2. Use of BM may thus facilitate more rapid generation of adoptive T cells with enhanced functionality. PMID- 19816192 TI - Selective transduction of mature DC in human skin and lymph nodes by CD80/CD86 targeted fiber-modified adenovirus-5/3. AB - In vivo targeting of dendritic cells (DC) represents an attractive alternative to currently apply ex vivo DC-based genetic tumor vaccination protocols. Finding the optimal vector for in vivo targeting of DC is important for such strategies. We, therefore, tested a panel of subgroup C/B chimeric and fiber-modified adenoviruses (Ads) for their relative capacity to transduce human DC. We made use of in vitro generated Langerhans cells, and of ex vivo human skin and melanoma draining lymph node derived DC. Of the tested viruses the C/B-chimeric adenovirus serotype 5 (Ad5)/3 virus most efficiently transduced in vitro generated Langerhans cells. In addition, Ad5/3 preferentially targeted mature myeloid DC from human skin and draining lymph node and transduced them at significantly higher frequencies than Ad5. In addition, Ad5/3 was more specific for mature human skin-derived CD1a+ CD83+ DC than the previously reported DC-transducing C/B chimeric vector Ad5/35, infecting less bystander cells. It was previously reported that Ad5/3 transduced human monocyte-derived DC by binding to the B7 molecules CD80 and CD86. High-efficiency transduction of mature skin-derived DC was similarly shown to be mediated through binding to CD80/CD86 and not to interfere with subsequent T-cell priming. We conclude that Ad5/3, in combination with DC-activating adjuvants, represents a promising therapeutic tool for the in vivo transduction of mature DC, and may be less likely to induce unwanted side effects such as immune tolerance through the infection of nonprofessional antigen presenting cells. PMID- 19816193 TI - Engineered interleukin-2 antagonists for the inhibition of regulatory T cells. AB - The immunosuppressive effects of CD4+ CD25 high regulatory T cells (Tregs) interfere with antitumor immune responses in cancer patients. Here, we present a novel class of engineered human interleukin (IL)-2 analogs that antagonizes the IL-2 receptor, for inhibiting regulatory T cell suppression. These antagonists have been engineered for high affinity to the alpha subunit of the IL-2 receptor and very low affinity to either the beta or gamma subunit, resulting in a signaling-deficient IL-2 analog that sequesters the IL-2 receptor alpha subunit from wild type IL-2. Two variants, "V91R" and "Q126T" with residue substitutions that disrupt the beta and gamma subunit binding interfaces, respectively, have been characterized in both a T cell line and in human primary Tregs. These mutants retain their high affinity binding to IL-2 receptor alpha subunit, but do not activate STAT5 phosphorylation or stimulate T cell growth. The 2 mutants competitively antagonize wild-type IL-2 signaling through the IL-2 receptor with similar efficacy, with inhibition constants of 183 pM for V91R and 216 pM for Q126T. Here, we present a novel approach to CD25-mediated Treg inhibition, with the use of an engineered human IL-2 analog that antagonizes the IL-2 receptor. PMID- 19816195 TI - Novel molecular stereotactic biopsy procedures reveal intratumoral homogeneity of loss of heterozygosity of 1p/19q and TP53 mutations in World Health Organization grade II gliomas. AB - We report a molecular stereotactic biopsy technique that combines histopathologic diagnosis with small sample size-adjusted molecular genetic analysis of low-grade gliomas that are ineligible for tumor resection. Loss of heterozygosity (LOH) of 1p/19q and TP53 mutations were analyzed in 1-mm tissue samples from 42 World Health Organization grade II gliomas (30 astrocytomas, 8 oligoastrocytomas, 4 oligodendrogliomas) using polymerase chain reaction-based microsatellite and sequence analysis. Alternating histological and molecular genetic evaluation within 1-mm steps at different sites within each tumor was performed to determine reproducibility of the results and the intratumoral distribution of the biomarkers. Multiple serial biopsies (range, 2-5 per tumor) taken from distinct intratumoral areas revealed concordant molecular genetic findings and homogeneous distribution of both biomarkers throughout 41 tumors. Contamination by nonneoplastic tissue could be recognized by corresponding histological evaluation and resulted in discordant LOH findings in 1 tumor. The frequency of LOH 1p/19q and TP53 mutations was consistent with the literature; these genetic alterations were found to be mutually exclusive. There was no biopsy-related morbidity. We conclude that determination of the LOH 1p/19q and TP53 status using this molecular stereotactic biopsy technique is safe and reliable in cases of unresectable gliomas. PMID- 19816194 TI - Nerve growth factor mediates mechanical allodynia in a mouse model of type 2 diabetes. AB - C57BLKS db/db (db/db) mice develop a neuropathy with features of human type 2 diabetic neuropathy. Here, we demonstrate that these mice develop transient mechanical allodynia at the early stage of diabetes. We hypothesized that nerve growth factor (NGF), which enhances the expression of key mediators of nociception (i.e. substance P [SP] and calcitonin gene-related peptide), contributes to the development of mechanical allodynia in these mice. We found that NGF, SP, and calcitonin gene-related peptide gene expression is upregulated in the dorsal root ganglion (DRG) of db/db mice before or during the period that they develop mechanical allodynia. There were more small- to medium-sized NGF immunopositive DRG neurons in db/db mice than in control db+ mice; these neurons also expressed SP, consistent with its role in nociception. Nerve growth factor expression in the hind paw skin was also increased in a variety of dermal cell types and nerve fibers, suggesting the contribution of a peripheral source of NGF to mechanical allodynia. The upregulation of NGF coincided with enhanced tropomyosin-related kinase A receptor phosphorylation in the DRG. Finally, an antibody against NGF inhibited mechanical allodynia and decreased the numbers of SP-positive DRG neurons in db/db mice. These results suggest that inhibition of NGF action is a potential strategy for treating painful diabetic neuropathy. PMID- 19816196 TI - A novel Caspr mutation causes the shambling mouse phenotype by disrupting axoglial interactions of myelinated nerves. AB - The neurological mouse mutation shambling (shm) exhibits ataxia and hindlimb paresis. Positional cloning of shm showed that it encodes contactin-associated protein (Caspr), which is required for formation of the paranodal junction in myelinated nerves. The shm mutation is a TT insertion in the Caspr gene that results in a frame shift and a premature stop codon at the COOH-terminus. The truncated Caspr protein that is generated lacks the transmembrane and cytoplasmic domains. Here, we found that the nodal/paranodal axoplasm of shm mice lack paranodal junctions and contain large mitochondria and abnormal accumulations of cytoplasmic organelles that indicate altered axonal transport. Immunohistochemical analysis of mutant mice showed reduced expression of Caspr, contactin, and neurofascin 155, which are thought to form a protein complex in the paranodal region; protein 4.1B, however, was normally distributed. The mutant mice had aberrant localization of voltage-gated ion channels on the axolemma of nodal/paranodal regions. Electrophysiological analysis demonstrated that the velocity of saltatory conduction was reduced in sciatic nerves and that the visual response was attenuated in the primary visual cortex. These abnormalities likely contribute to the neurological phenotype of the mutant mice. PMID- 19816197 TI - Decreased cystatin C immunoreactivity in spinal motor neurons and astrocytes in amyotrophic lateral sclerosis. AB - Cystatin C (CC), a cysteine protease inhibitor involved in protein degradation, is a marker of Bunina bodies in lower motor neurons in amyotrophic lateral sclerosis (ALS). TAR-DNA binding protein-43 (TDP-43)-immunoreactive inclusions are also histological hallmarks of ALS but whether CC is found in motor neurons with or without TDP-43-positive inclusions in ALS is not known. To determine whether inclusion body formation affects cytoplasmic CC immunoreactivity, we examined spinal cords from 9 ALS patients and 12 control subjects by immunohistochemistry. Most anterior horn cells (AHCs) showed moderate to intense immunoreactivity in controls, whereas CC immunoreactivity was markedly decreased in AHCs in ALS cases. The proportion of CC-immunolabeled AHCs was reduced regardless of whether they contained Bunina bodies. In contrast, the proportion of CC-immunolabeled AHCs was significantly reduced in those with TDP-43 inclusions. Cystatin C immunoreactivity of astrocytes in the spinal gray matter and white matter in ALS was significantly decreased compared with controls. These findings suggest that the formation of TDP-43 inclusions, but not of Bunina bodies, may be linked to the content of CC in spinal motor neurons and that perturbations in endogenous levels of CC in neuronal and glial cells may be part of the neurodegenerative processes in ALS. PMID- 19816198 TI - Mitochondrial ferritin in the substantia nigra in restless legs syndrome. AB - Restless legs syndrome (RLS) is a neurological disorder that is thought to involve decreased iron availability in the brain. Iron is required for oxidative metabolism and plays a critical role in redox reactions in mitochondria. The recent discovery of mitochondrial ferritin (FtMt) provided the opportunity to identify a potential correlation between iron and mitochondrial function in RLS. Human substantia nigra (SN) and putamen autopsy samples from 8 RLS cases and 8 controls were analyzed. Mitochondrial ferritin levels in RLS SN tissue homogenate samples assessed by immunoblots had more FtMt than control samples (p < 0.01), whereas there were no significant differences in FtMt in the putamen samples. By immunohistochemistry, neuromelanin-containing neurons in the SN were the predominant cell type expressing FtMt. Staining in neurons in RLS samples was consistently greater than that in controls. Cytochrome c oxidase staining, which reflects numbers of mitochondria, showed a similar staining pattern to that of FtMt, whereas there was less immunostaining in the RLS cases for cytosolic H ferritin. These results suggest that increased numbers of mitochondria in neurons in RLS and increased FtMt might contribute to insufficient cytosolic iron levels in RLS SN neurons; they are consistent with the hypothesis that energy insufficiency in these neurons may be involved in the pathogenesis of RLS. PMID- 19816199 TI - Nuclear contour irregularity and abnormal transporter protein distribution in anterior horn cells in amyotrophic lateral sclerosis. AB - The nucleocytoplasmic transport system is essential for maintaining cell viability; transport of proteins and nucleic acids between the nucleus and the cytoplasm occurs through nuclear pore complexes (NPCs). In this study, we examined the immunohistochemical distribution of the major protein components of NPCs, Nup62, Nup88, and Nup153, in spinal cords from controls and patients with sporadic or familial amyotrophic lateral sclerosis (SALS or FALS) and its mouse model. In control subjects, immunolabeling on the nuclear envelopes of anterior horn cells (AHCs) was invariably smooth and continuous, whereas in SALS and FALS patients, the AHCs predominantly showed irregular nuclear contours. Double immunofluorescence staining demonstrated that in SALS patients, importin-beta immunoreactivity was absent in the nuclei in a subset of AHCs; in these cells, Nup62 immunolabeling of nuclear membrane was invariably irregular, suggesting that there was dysfunctional nucleocytoplasmic transport in those AHCs. In the mouse model, Nup62-immunolabeled AHCs with irregular nuclear contours were predominant as early as the presymptomatic stage and the contours became progressively discontinuous along with disease development. Together, these observations suggest that dysfunctional nucleocytoplasmic transport may underlie the pathogenesis of ALS. PMID- 19816200 TI - Bone morphogenetic protein 4 and its receptors are expressed in the leptomeninges and meningiomas and signal via the Smad pathway. AB - The roles of bone morphogenetic proteins (BMPs) and their receptors (BMPRs) in meningioma biology are not known. In this study, frozen tissues from 26 World Health Organization Grades I to III meningiomas were analyzed by Western blot for BMP-2/4, BMPR IA, and BMPR II, and activation of downstream p-Smad1, p38 mitogen activated protein kinase (MAPK), and p44/42 MAPK signaling molecules. Sections from 20 normal leptomeninges, 2 arachnoid cysts, and 51 meningiomas were analyzed for BMP-4 and p44/42 MAPK by immunohistochemistry. Primary meningioma cultures from 11 meningiomas were treated with BMP-4 and evaluated for cell proliferation and signaling pathway activation. Conditioned media from 7 cultures were analyzed for BMP-4 by ELISA. Bone morphogenetic protein 4 was variably detected in adult leptomeninges but was detected in 89% or 84% of Grade I meningiomas and in 60% of Grade II meningiomas by Western blot and immunohistochemistry, respectively. Bone morphogenetic protein receptors IA and II were detected in leptomeninges and in all meningiomas studied, and activated Smad1 was detected in all meningiomas studied. Bone morphogenetic protein 4 stimulated meningioma cell proliferation and phosphorylation/activation of Smad1 but not p38 MAPK or p44/42 MAPK in vitro, and it was detected in conditioned media from 4 of 7 cultures. These findings suggest that BMP-4 and BMPRs may play autocrine/paracrine roles and interact with other transforming growth factor-beta superfamily members in regulating meningioma growth and differentiation. PMID- 19816201 TI - Transactivation response DNA-binding protein 43 microvasculopathy in frontotemporal degeneration and familial Lewy body disease. AB - We describe novel transactivation response DNA-binding protein of 43 kd (TDP-43) positive structures in the brains of patients with frontotemporal lobar degeneration with ubiquitin-positive inclusions and familial Lewy body disease. The TDP-43 immunohistochemistry revealed small round structures closely associated with small blood vessels. By immunoelectron microscopy, these TDP-43 positive structures were unmyelinated cell processes located adjacent to and sometimes enclosed by the capillary basal lamina. Some processes protruded from outside of the vascular basal lamina to a position beneath the basal lamina. The processes contained 10- to 17-nm-diameter straight filaments or filaments coated with granular material similar to those described in neurites in frontotemporal lobar degeneration with ubiquitin-positive inclusions and other disorders. In some of the abnormal structures, electron-dense material formed paracrystalline arrays composed of TDP-43. The inclusions were variably positive by immunostaining for the small heat shock protein alphaB-crystallin and less often glial fibrillary acidic protein. Bundles of astrocytic glial fibrils characteristic of reactive astrocytes were often found in proximity, but glial fibrils were negative for TDP-43. These data suggest that these processes are astrocytic end-feet with abnormal TDP-43 fibrillary inclusions. The significance of this novel TDP-43 microvasculopathy on blood-brain barrier integrity warrants further investigation. PMID- 19816202 TI - Occipital nerve stimulator placement under general anesthesia: initial experience with 5 cases and review of the literature. AB - Anesthesiologists support nerve stimulator insertion procedures, including occipital nerve stimulator placement for refractory headache disorders. Sedation during these cases can be challenging on account of variable surgical stimuli and surgery positioning that contribute to neck flexion, potentially compromising the airway. Greater patient comfort and safety may be found in performing permanent occipital stimulator placement procedures entirely under general anesthesia, assuming that appropriate stimulation patterns can be achieved in patients who are unable to provide intraoperative feedback. The purpose of this study is to describe our initial experience with occipital nerve stimulator placement performed entirely under general anesthesia and the resulting stimulation patterns, and to review the medical literature regarding the anesthetic techniques used during these novel neurosurgical procedures. After institutional review board approval, we reviewed the records of 5 patients who underwent permanent occipital nerve stimulator placement under general anesthesia. Appropriateness of the postoperative stimulation patterns was noted in addition to complications. The medical literature was searched for occipital stimulation surgery studies that also described the anesthetic technique. We found that all 5 patients underwent uncomplicated general anesthetics. Postoperative occipital stimulation was nonpainful and symmetrical for all. The literature search provided little information on the anesthetic technique; most procedures were performed at least in part under local anesthesia with sedation. On the basis of this small case series, we conclude that the occipital nerve stimulator systems can be successfully placed under general anesthesia while still achieving the desired occipital region stimulation. Further studies are needed to correlate occipital nerve stimulator placement under general anesthesia and long-term headache control. PMID- 19816203 TI - Application of the FlexiForce contact surface force sensor to continuous extraocular compression monitoring during craniotomy for cerebral aneurysms. AB - The aims of this study were to introduce our newly developed device equipped with a contact surface force sensor (FlexiForce) for monitoring extraocular compression continuously, and to illustrate its potential clinical application using this device in patients undergoing uncomplicated frontotemporal or bifrontal craniotomy for surgical clipping of unruptured anterior circulation aneurysms. In a pilot study with volunteers, we determined the critical force of 100 gf to cause painful ocular sensation. Then we performed the bilateral extraocular force measurements in 15 patients undergoing uncomplicated frontotemporal or bifrontal craniotomy for surgical clipping of unruptured anterior circulation aneurysms. Extraocular force increased immediately after retraction of the flap, increased to 144+/-26 gf (mean+/-SD) during lower craniotomy close to the orbit, was maintained at 91+/-18 gf during microsurgery, and returned close to baseline at 24+/-14 gf after restoration of skin flap retraction. Such changes were observed only on the surgical side in frontotemporal craniotomy. Abnormal increase in extraocular force was effectively reduced by placing a real-time digital panel meter to warn surgeons to avoid excessive skin flap retraction during the surgical procedure. In conclusion, this new tool may allow us to monitor the external forces that can be applied intraoperatively to the ocular globe in the supine position. PMID- 19816204 TI - Intraoperative monitoring of cerebral microcirculation and oxygenation--a feasibility study using a novel photo-spectrometric laser-Doppler flowmetry. AB - BACKGROUND: The present study assesses the utility of a novel invasive device (O2C-, oxygen-to-see-device) for intraoperative measurement of the cerebral microcirculation. CO2 vasoreactivity during 2 different propofol concentrations was used to investigate changes of capillary venous cerebral blood flow (rvCBF), oxygen saturation (srvO2), and hemoglobin concentration (rvHb) during craniotomy. METHODS: Thirty-four patients were randomly assigned to a low propofol (4 mg/kg/h) versus a high propofol (6 mg/kg/h) group. A fiberoptic probe was applied on the cortex next to the surgical site. Measurements were performed during lower (35 mm Hg) and higher (45 mm Hg) levels of partial pressure of carbon dioxide (paCO2). Arterio-venous difference in oxygen concentration (avDO2) and approximated cerebral metabolic rate of oxygen (aCMRO2) were calculated for each paCO2 state. Linear models were fitted to test changes of end points in response to paCO2 and propofol concentration. RESULTS: In comparison to the lower levels of paCO2, higher levels of paCO2 increased rvCBF (P<0.001), and srvO2 (P=0.002). RvHb remained unchanged during measurements (P=0.325). Calculated avDO2 decreased with increasing paCO2 (P<0.001), whereas aCMRO2 did not change during the study (P=0.999). Propofol concentration had no effect on measured or calculated end points. CONCLUSIONS: Increase of rvCBF by paCO2 indicates a preserved CO2 reactivity independent of propofol anesthesia. The consecutive rise in srvO2 implies enhanced oxygen availability due to vasodilatation. Unchanged rvHb represents constant venous hemoglobin concentration. As expected, calculated avDO2 decreases with increased paCO2, whereas aCMRO2 remains unchanged. Despite the promising technical approach, the technology needs validation and further investigation for usage during neurosurgery. PMID- 19816205 TI - Effect of the combination of mannitol and ringer acetate or hydroxyethyl starch on whole blood coagulation in vitro. AB - Mannitol is administered to decrease the intracranial pressure and to improve surgical conditions during craniotomy. Simultaneously a crystalloid infusion is always given and sometimes hydroxyethyl starch (HES) is indicated for intravascular volume replacement. As normal coagulation profile is required during craniotomy, we aimed at determining the effect of mannitol with or without HES or Ringer acetate on blood coagulation in this randomized cross-over in vitro study. Blood samples were withdrawn from 10 volunteers. From whole blood we prepared 10 vol.% and 20 vol.% dilutions of mannitol (15% Mannitol) alone, mannitol and Ringer acetate, and mannitol and HES 130/0.4 (Voluven) at a ratio of 1:1. Blood samples were analyzed by modified thromboelastometry. Coagulation parameters: clotting time, clot formation time, and maximum clot firmness (MCF), were registered. Clot formation time was prolonged in all dilutions compared with control (P<0.05). MCF decreased in all dilutions compared with control (P<0.05). MCF in 20 vol.% dilution of mannitol with HES was lower than MCF in the corresponding dilution with Ringer acetate (P<0.05). Fibrinogen-dependent MCF in 10 vol.% dilution of mannitol with HES was lower than MCF in the corresponding dilution with Ringer acetate (P<0.05). We conclude that mannitol in combination with HES 130/0.4 impairs clot propagation and clot strength in vitro. Fibrin clot strength impairment is more pronounced when mannitol is combined with HES than Ringer acetate. Our findings indicate that HES in combination with mannitol should be avoided whenever a disturbance in hemostasis is suspected during craniotomy. PMID- 19816206 TI - Enlarged and prominent nucleus may be indicative of tetraploidy: a laboratory study of a rare near-tetraploidy in a child patient with acute myelogenous leukemia AML-M4. AB - SUMMARY: Near-tetraploidy is a rare cytogenetic abnormality in myelocytic malignancies in children and its significance is unknown. To investigate the pathologic characteristics of a near-tetraploidy in a child with acute myelogenous leukemia (AML-M4), bone marrow smears were prepared for morphologic analysis. Bone marrow samples were collected at presentation for flow cytometry, prepared by short-term (24 h) unstimulated culture and R-banding for conventional cytogenetic assay. We have performed a multifactorial analysis of the laboratory test results. In this case, the chromosomal analysis (R-banding) demonstrated a near-tetraploidy. Combined with morphologic and immunophenotypic results, the diagnosis was established as acute myelogenous leukemia (AML-M4). Near tetraploidy is an uncommon cytogenetic finding, and the experience of this case further emphasizes the importance of the laboratory diagnostic methods. PMID- 19816207 TI - Detection and clinical relevance of BCR-ABL fusion gene in childhood T-lineage acute lymphoblastic leukemia: a report on 4 cases. AB - The bcr-abl rearrangement has rarely been reported in T-lineage acute lymphoblastic leukemia and the clinical significance of this translocation is currently unknown. We screened 28 children with T-lineage acute lymphoblastic leukemia at diagnosis by reverse transcription polymerase chain reaction for major and minor break point regions of bcr-abl fusion gene. Four out of 28 patients (14.2%) were bcr-abl positive for the minor breakpoint transcript. One of these patients was refractory to therapy, whereas the other 3 relapsed on therapy. PMID- 19816208 TI - Comparison of safety and effectiveness of two different transfusion rates in children with severe anemia. AB - We compared the safety and efficacy of 2 transfusion regimens in children with severe anemia (hemoglobin <5 g/dL) and without overt signs of congestive heart failure requiring transfusion of packed red blood cells (PRBCs). Forty-three patients were randomly divided into 2 groups: group A and group B. The transfusion regimens consisted of continuous infusion of PRBC at a rate of 1 mL/kg/h for group A and 3 mL/kg/h for group B. The patients were closely monitored for any clinical signs of heart failure throughout transfusion. Heart and respiratory rate, and arterial blood pressure were measured hourly for 6 hours during transfusion. None of the patients developed any signs of cardiac failure during or after the transfusion. The mean heart rate, respiratory rate, diastolic and systolic blood pressure were similar in both groups throughout transfusion. Group A needed significantly more PRBC units than group B to attain the same hemoglobin increase. Transfusion of PRBC at a rate of 3 mL/kg/h, and at a rate of 1 mL/kg/h, is a safe regimen for children with severe anemia of gradual onset requiring transfusion therapy. PMID- 19816209 TI - Erythrocyte transfusions and serum prohepcidin levels in premature newborns with anemia of prematurity. AB - Hepcidin is a regulatory peptide hormone acts by limiting intestinal iron absorption and promoting iron retention. Determining the level of hepcidin in anemia of prematurity might be important in preventing iron overload. This study aimed to determine serum levels of prohepcidin in newborns with anemia of prematurity, to assess the effect of a single erythrocyte transfusion on serum prohepcidin levels, and to determine the possible relationships between prohepcidin levels and serum iron and complete blood count parameters. Nineteen premature newborns with anemia of prematurity who had been treated with erythrocyte transfusions were included in this study. Just before, and 48 hours after, each transfusion, venous blood samples were collected from patients. Serum prohepcidin levels before and after erythrocyte transfusion were 206.5+/-27.3 and 205.7+/-47.1 ng/mL, respectively; no statistically significant differences were found. No significant differences existed before or after transfusion regarding serum total iron and ferritin levels, iron-binding capacity, or mean corpuscular hemoglobin concentration. No significant correlations existed between serum prohepcidin levels and other parameters, either before or after transfusions. Our results showed that there were no statistically significant differences between serum prohepcidin levels before and after a single erythrocyte transfusion in premature newborns. PMID- 19816210 TI - A novel mutation in a Fijian boy with Shwachman-Diamond syndrome. AB - Shwachman-Diamond syndrome (SDS) is an autosomal recessive disorder characterized by pancreatic insufficiency, bone marrow dysfunction, and metaphyseal chondrodysplasia. SDS is associated with mutations in the Shwachman-Bodian Diamond Syndrome gene, with 90% of reported mutations in exon 2. We present a Fijian boy with SDS who has a novel A>G substitution in exon 1 of the Shwachman Bodian-Diamond Syndrome gene that has not been reported in the literature. This patient's unique clinical course includes the presence of a cleft lip and episodic hypoglycemia. SDS lacks a clear genotype-phenotype correlation, as is showed by the heterogeneity in its clinical presentation. PMID- 19816211 TI - Effects of creatine supplementation during resistance training on myosin heavy chain (MHC) expression in rat skeletal muscle fibers. AB - The purpose of this study was to utilize a rodent model to test the hypothesis that creatine (Cr) supplementation during resistance training would influence the pattern of slow-twitch muscle myosin heavy chain (MHC) isoforms expression. Male Wistar rats (2-3 months old, 250-300 g) were divided into 4 groups: Nontrained without creatine supplementation (CO), nontrained with creatine supplementation (CR), trained without creatine supplementation (TR), and trained with creatine supplementation (TRCR). TR and TRCR groups were submitted to a resistance training program for 5 weeks (5 days/week) for morphological and biochemical analysis of the soleus muscle. Weightlifting exercise involved jump sessions into water, carrying progressive overload equivalent to percentage of body weight. CR and TRCR groups were given creatine at 0.5 g/kg(-1)/d(-1). Both Cr supplementation and resistance training alone or associated did not result in significant alterations (p > 0.05) in body weight gain, food intake, and muscle weight in the CR, TR and TRCR groups compared to the CO group. Also compared to the CO group, the CR group showed a significant (p < 0.02) increase in MHCI content and a reduction in MHCII; inversely, the TR group increased the MHCII content and reduced MHCI (p < 0.02). When combined, both creatine and resistance training did not promote significant (p > 0.05) changes in MHC content of the TRCR group compared to the CO group. The data show that Cr supplementation provides a potential action to abolish the exercise-induced MHC isoform transitions from slow to fast in slow-twitch muscle. Thus, Cr supplementation might be a suitable strategy to maintaining a slow phenotype in slow muscle during resistance training, which may be favorable to maintenance of muscle oxidative capacity of endurance athletes. PMID- 19816212 TI - Research note: effect of load reductions over consecutive sets on repetition performance. AB - When performing consecutive sets of a resistance exercise, maintaining performance within a specified repetition range has been shown to be essential for achieving muscular adaptations conducive to different training goals. However, maintaining repetition performance can be difficult when using shorter rest intervals between sets (i.e., < or = 1 min), which may require load reductions. Therefore, the purpose of the current study was to compare repetition performance when keeping the load constant or reducing the load by different percentages during a lower-body workout. Ten repetition maximum (10RM) loads were established for the back squat, leg curl, and leg extension exercises. Subjects performed 4 workouts under the following load conditions: (a) constant load for all sets, (b) 5% load reduction after each set, (c) 10% load reduction after each set, and (d) 15% load reduction after each set. Pairwise comparisons indicated that, when averaged across sets, significantly fewer repetitions were accomplished for the back squat and leg curl within the constant condition vs. the 15% condition (p < 0.05). Conversely, for the leg extension, there were no significant differences in the repetitions accomplished between conditions (p > 0.05). Pairwise comparisons also indicated that, when averaged across exercises, significantly fewer repetitions were accomplished for set 3 within the constant, 5%, and 10% conditions vs. the 15% condition (p < 0.05). In summary, the back squat and leg curl required 15% load reductions per set to maintain repetition performance. Conversely, load reductions were not necessary for the leg extension. PMID- 19816213 TI - Low-calorie energy drink improves physiological response to exercise in previously sedentary men: a placebo-controlled efficacy and safety study. AB - Energy drink use has grown despite limited research to support efficacy or safety and amid concerns when combined with exercise. The purpose of this study was to assess the effects of 10 weeks of once-daily energy drink consumption or energy drink consumption with exercise on measures of body composition, cardiorespiratory fitness, strength, mood, and safety in previously sedentary males. Thirty-eight males were randomly assigned to energy drink + exercise (EX A), energy drink (NEX-A), placebo + exercise (EX-B), or placebo (NEX-B). All participants consumed 1 drink per day for 10 weeks; EX-A and EX-B participated in 10 weeks of resistance and endurance exercise. Testing was performed before (PRE) and after (POST) the 10-week intervention. No significant (p > 0.05) changes were observed for body composition, fitness, or strength in NEX-A; however, significantly greater decreases in fat mass and percentage body fat and increases in VO2peak were observed in EX-A versus EX-B. Ventilatory threshold (VT), minute ventilation, VO2 at VT, and power output at VT improved significantly PRE to POST in EX-A but not in EX-B or nonexercising groups. Clinical markers for hepatic, renal, cardiovascular, and immune function, as determined by PRE and POST blood work revealed no adverse effects in response to the energy drink. Mood was not affected by energy drink use. Absent energy restriction or other dietary controls, chronic ingestion of a once-daily low-calorie energy drink appears ineffective at improving body composition, cardiorespiratory fitness, or strength in sedentary males. However, when combined with exercise, preworkout energy drink consumption may significantly improve some physiological adaptations to combined aerobic and resistance training. PMID- 19816214 TI - Static stretching does not impair performance in active middle-aged adults. AB - Recent investigations with young, healthy adult subjects suggest that static stretching before activity decreases performance and should, therefore, be avoided. The purpose of this study was to assess the effects of an acute static stretching protocol on balance and jump/hop performance in active middle-aged adults. Ten subjects (6 men and 4 women aged 40-60 yr) from a martial arts school volunteered to take part in this research study. This was a repeated measures design. Subjects who stretched for 10 minutes using a 30-second hold during 1 session sat quietly for 10 minutes during the alternate session. Sessions were randomly assigned. The following dependent variables were compared: Dynamic Stability Index (DSI) for single-leg dynamic balance (smaller DSI = improved balance); distances for broad jump, single hop, triple hop, and crossover hop; elapsed time for a 6-m timed hop. Group means for balance were significantly different between the stretch and no-stretch conditions (3.5 +/- 0.7 vs. 4.3 +/- 1.4 DSI, respectively; p < 0.05). No significant differences were found between the group means of the stretch and no-stretch conditions for the dependent measures of broad jump, single hop, triple hop, crossover hop, and 6-m timed hop performance. Ten minutes of acute static stretching enhances dynamic balance and does not affect jump/hop performance in active middle-aged adults. Static stretching should be included before competition and before exercise in fitness programs of active middle-aged adults. PMID- 19816215 TI - Effect of preseason concurrent muscular strength and high-intensity interval training in professional soccer players. AB - This study examined the effect of concurrent muscular strength and high-intensity running interval training on professional soccer players' explosive performances and aerobic endurance. Thirty-nine players participated in the study, where both the experimental group (EG, n = 20) and control group (CG, n = 19) participated in 8 weeks of regular soccer training, with the EG receiving additional muscular strength and high-intensity interval training twice per week throughout. Muscular strength training consisted of 4 sets of 6RM (repetition maximum) of high-pull, jump squat, bench press, back half squat, and chin-up exercises. The high intensity interval training consisted of 16 intervals each of 15-second sprints at 120% of individual maximal aerobic speed interspersed with 15 seconds of rest. EG significantly increased (p < or = 0.05) 1RM back half squat and bench press but showed no changes in body mass. Within-subject improvement was significantly higher (p < or = 0.01) in the EG compared with the CG for vertical jump height, 10-m and 30-m sprint times, distances covered in the Yo-Yo Intermittent Recovery Test and maximal aerobic speed test, and maximal aerobic speed. High-intensity interval running can be concurrently performed with high load muscular strength training to enhance soccer players' explosive performances and aerobic endurance. PMID- 19816216 TI - Endocrine response patterns to acute unilateral and bilateral resistance exercise in men. AB - Rehabilitation programs and research experiments use single-arm protocols in which the contralateral arm is not functional or used as a control limb. This study was interested in determining the hormonal signal impacts of such one- versus two-arm exercise responses that might have an impact on adaptational changes with training. The purpose was to examine the acute hormonal responses to a unilateral and a bilateral upper-body resistance exercise (RE) protocol. A balanced randomized treatment intervention with series time frame for blood collections before and after exercise was used as the basic experimental design. Ten recreationally resistance trained men (18-25 years, 20.4 +/- 1.2 years, 175.6 +/- 4.5 cm, 81.7 +/- 9.3 kg) gave informed consent to participate in the investigation. Each subject performed unilateral (dominant arm only) and bilateral upper-body RE protocol separated by 1 week in a balanced randomized fashion. The RE protocol consisted of 3 sets of 10 repetitions of 5 different dumbbell upper-body exercises at 80% of 1-repetition maximum, and blood samples were obtained before and 5, 15, and 30 minutes immediately postexercise (IP). Blood was obtained and analyzed for lactate, immunoreactive growth hormone (iGH), cortisol (C), total testosterone (T), and insulin concentrations. Total volume of work also was determined for the 2 exercise sessions. Total volume of work performed during the unilateral protocol was 52.1% of that for the bilateral protocol. Both RE protocols elicited a significant (p < or = 0.05) increase in lactate and iGH, but the increase for the bilateral condition was significantly greater. Cortisol decreased significantly during recovery for the unilateral condition. Testosterone was not affected by either protocol. Insulin was significantly increased at IP and 5 minutes postexercise for both conditions.These results indicate that the hormonal responses to dominant-arm unilateral RE is blunted compared to that for bilateral RE. This differential endocrine response is likely a result of the difference in volume between the protocols. It is important to pay attention to the amount of muscle mass utilized in a resistance exercise protocol to optimize endocrine signaling. PMID- 19816217 TI - Relationship between off-ice testing variables and on-ice speed in women's collegiate synchronized figure skaters: implications for training. AB - The purpose of the current investigation was to identify any existing relationships between off-ice performance measures and on-ice performance quantified by speed and acceleration. Twenty-seven women (age 19 +/- 1 year; body mass (59.5 +/- 6.8 kg; height 164.6 +/- 6.35 cm; body fat 23.2 +/- 3.9%) who were collegiate synchronized figure skaters volunteered for the investigation. To examine the relationship between off-ice performance and on-ice speed and acceleration, collegiate synchronized skaters were evaluated on various performance tests over a 1-week period. Off-ice tests completed were peak torque for hip abduction and adduction, 40-yard sprint, vertical jump height, 30-second slide board stride count, and a 1-RM (repetition maximum) squat. On-ice tests included a timed single lap sprint, 4.5-minute (duration of long program) lap count, and an approximately 16.5-m (18-yard blue line to blue line) timed acceleration. Significance was set at P < or = 0.05. This study showed 3 primary findings: (a) slide board stride count was the single best predictor for both single lap on-ice speed and acceleration accounting for 53.5% (adjusted R2 value) of the variance in the single lap test and 42.5% (adjusted R2 value) of the variance in acceleration times; (b) vertical jump height test was the second best predictor for both the single lap test and on-ice acceleration accounting for 36.6% and 39.9% (adjusted R2 values) of the variance in times recorded, respectively; and (c) the best combined predictors for the single lap speed test were slide board stride count and 40-yard dash (R2 = 0.675), whereas the best combined predictors for on-ice acceleration were slide board stride count and vertical jump height test (R2 = 0.571). Conditioning for synchronized skaters to enhance performance of on-the-ice speed and acceleration should include slide board training implementation of plyometric and linear speed training while developing and maintaining 1-RM strength to support power capabilities. PMID- 19816218 TI - Effect of an acute bout of whole body vibration exercise on muscle force output and motor neuron excitability. AB - The purpose of the current investigation was to assess the effect of an acute bout of whole body vibration (WBV) exercise on muscle force output and motor neuron excitability. Nineteen recreationally trained college-aged males were randomly assigned to a WBV (n = 10) or a sham (S, n = 9) group. The WBV group completed a series of static, body weight squats on a vibrating platform at 30 Hz and an amplitude of approximately 3.5 mm (vertical), whereas the S group performed the same series of exercises but without vibration. Measurements were performed before (Pre) and then immediately post-exercise (Imm Post), 8 minutes post-exercise (8-Min Post), or 16 minutes post-exercise (16-Min Post) during 3 different testing sessions. The measurements involved a ballistic isometric maximum voluntary contraction (MVC) of the triceps surae muscle complex and electrical stimulation of the tibial nerve for assessment of motor neuron excitability by analyzing H-reflex and M-wave responses (H(max)/M(max) ratio). Electromyography was also obtained from the triceps surae muscle complex during the MVCs. The WBV group significantly (p < or = 0.05) increased peak force at Imm Post (9.4%) and 8-Min Post (10.4%). No significant change in peak force was observed in the S group. No significant changes were observed in either group for average integrated EMG, H(max)/M(max) ratio, or rate of force development at Imm Post, 8-Min Post, or 16-Min Post. The results from this investigation indicate that an acute bout of static, body weight squat exercises, combined with WBV, increases muscle force output up to 8 minutes post-exercise. However, this increase in muscle force is not accompanied by a significant increase in motor neuron excitability or muscle activation. Thus, it is plausible to use WBV as a method for acute increase in muscle force output for athletes immediately before competition. PMID- 19816219 TI - Kinetic and kinematic differences between squats performed with and without elastic bands. AB - The purpose of this investigation was to compare kinetic and kinematic variables between squats performed with and without elastic bands equalized for total work. Ten recreationally weight trained males completed 1 set of 5 squats without (Wht) and with (Band) elastic bands as resistance. Squats were completed while standing on a force platform with bar displacement measured using 2 potentiometers. Electromyography (EMG) was obtained from the vastus lateralis. Average force time, velocity-time, power-time, and EMG-time graphs were generated and statistically analyzed for mean differences in values between the 2 conditions during the eccentric and concentric phases. The Band condition resulted in significantly higher forces in comparison to the Wht condition during the first 25% of the eccentric phase and the last 10% of the concentric phase (p < or = 0.05). However, the Wht condition resulted in significantly higher forces during the last 5% of the eccentric phase and the first 5% of the concentric phase in comparison to the Band condition. The Band condition resulted in significantly higher power and velocity values during the first portion of the eccentric phase and the latter portion of the concentric phase. Vastus lateralis muscle activity during the Band condition was significantly greater during the first portion of the eccentric phase and latter portion of the concentric phase as well. This investigation indicates that squats equalized for total work with and without elastic bands significantly alter the force-time, power-time, velocity-time, and EMG-time curves associated with the movements. Specifically, elastic bands seem to increase force, power, and muscle activity during the early portions of the eccentric phase and latter portions of the concentric phase. PMID- 19816220 TI - Elastic band prediction equations for combined free-weight and elastic band bench presses and squats. AB - Elastic bands added to traditional free-weight techniques have become a part of suggested training routines in recent years. Because of the variable loading patterns of elastic bands (i.e., greater stretch produces greater resistance), it is necessary to quantify the exact loading patterns of bands to identify the volume and intensity of training. The purpose of this study was to determine the length vs. tension properties of multiple sizes of a set of commonly used elastic bands to quantify the resistance that would be applied to free-weight plus elastic bench presses (BP) and squats (SQ). Five elastic bands of varying thickness were affixed to an overhead support beam. Dumbbells of varying weights were progressively added to the free end while the linear deformation was recorded with each subsequent weight increment. The resistance was plotted as a factor of linear deformation, and best-fit nonlinear logarithmic regression equations were then matched to the data. For both the BP and SQ loading conditions and all band thicknesses tested, R values were greater than 0.9623. These data suggest that differences in load exist as a result of the thickness of the elastic band, attachment technique, and type of exercise being performed. Facilities should adopt their own form of loading quantification to match their unique set of circumstances when acquiring, researching, and implementing elastic band and free-weight exercises into the training programs. PMID- 19816221 TI - Muscle architecture and optimum angle of the knee flexors and extensors: a comparison between cyclists and Australian Rules football players. AB - The purpose of this study was to investigate differences in optimum angle of peak torque (knee extensors and flexors) and muscle architecture (vastus lateralis) between 9 cyclists and 9 Australian Rules football (ARF) players. The angles of peak torque of the ARF players were significantly (p < 0.05) greater during knee extension (70.8 +/- 3.5 vs. 66.6 +/- 5.9 degrees) and smaller during knee flexion (26.2 +/- 2.9 vs. 32.3 +/- 3.8 degrees) compared with the cyclists. The ARF players had significantly (p < 0.05) smaller pennation angles (19.3 +/- 2.0 vs. 24.9 +/- 2.5 degrees) and longer fascicle lengths (7.9 +/- 0.7 vs. 6.2 +/- 0.8 cm) in comparison with the cyclists. There were no significant differences between groups regarding muscle thickness or peak torque ratios between the quadriceps and hamstrings. Muscle architectural changes associated with resistance strength training need to be investigated so that the effects of training on architecture and functional performance can be determined. PMID- 19816222 TI - The history of middle cranial fossa approach to the cerebellopontine angle. AB - OBJECTIVES: To investigate the historical origins of the current middle fossa (MF) approach to the cerebellopontine angle (CPA). METHODS: A review of more than 30 original articles from the 1880 s to the early 1960s that document the evolution of the MF approach. Historically important journal articles and book chapters in various languages were supplemented by interviews with surgeons instrumental in the adaptation of this method to modern microsurgical neurotology. RESULTS: The inspiration for the current MF approach could be traced back to the late 19th century extradural subtemporal surgeries for the trigeminal neuralgia performed by Krause and Hartley and the contemporary surgeries performed by Ernst von Bergmenn for treatment of otogenic temporal bone infections. The first case of MF approach to CPA for vestibular nerve section was reported by RH Parry in 1904. In the mid-20th century, otologists found numerous applications for this approach, which gained popularity at the hands of William House. Although his initial MF operations aimed to decompress the internal auditory canal for cochlear otosclerosis, House soon realized the versatility of this approach and performed his first MF acoustic neuroma surgery in 1961. CONCLUSION: The current MF approach has its roots in operations for trigeminal neuralgia and otologic surgeries for palliation of temporal bone infections performed in the late 19th century. The earliest reported MF approach to the CPA dates back to 1904 and its application to microsurgical exposure of the internal auditory canal to the late 1950s. PMID- 19816223 TI - Electroconvulsive therapy in a cochlear implant patient. AB - OBJECTIVE: To report the first use of electroconvulsive therapy (ECT) in a patient with a cochlear implant (CI). STUDY DESIGN: Clinical Capsule Report. SETTING: University hospital. PATIENT: : A 17-year-old boy who underwent Nucleus 22 cochlear implantation in 1995 presented with delirious mania in 2009. Aggressive pharmacologic management was ineffective, and ECT was recommended due to the potentially lethal nature of his psychiatric illness. INTERVENTIONS: After careful consideration by a multidisciplinary team, unilateral ECT on the side opposite the CI without removal of the device was recommended. Electroconvulsive therapy was performed on hospital Days 20 and 21. Integrity testing of the CI was performed on hospital Day 38. MAIN OUTCOME MEASURES: Subjective and objective assessment of cochlear implant functioning and response to ECT. RESULTS: Electroconvulsive therapy was well tolerated and contributed to alleviation of presenting symptoms. The patient used his CI without subjective degradation of performance. Integrity testing of the CI after ECT confirmed proper functioning of the device. CONCLUSION: This is the first report of ECT in a patient with CI. Unilateral ECT was performed contralateral to the CI without subjective or objective decline in performance. This Clinical Report motivates further study regarding the use of ECT in CI patients. PMID- 19816224 TI - Ocular vestibular evoked myogenic potentials to bone-conducted vibration in vestibular schwannomas. AB - BACKGROUND: Vestibular schwannoma (VS) arises from either the superior or the inferior vestibular nerve and causes vestibular dysfunction to various degrees. Recently, ocular vestibular evoked myogenic potentials to bone-conducted vibration (oVEMPs to BCV) have attracted much interest as a new clinical test for otolith-ocular pathway function. Because it is unclear whether the oVEMPs to BCV primarily originate from activation of the superior or the inferior vestibular nerve, the results in patients with VS might enlighten us concerning the origin of oVEMPs to BCV. OBJECTIVE: To compare the results of 3 clinical tests for vestibular function in patients with VS: 1) oVEMPs to BCV; 2) cervical vestibular evoked myogenic potentials to air-conducted sound (cVEMPs to ACS), which reflect the function of inferior vestibular nerve; and 3) caloric test, which reflect the function of superior vestibular nerve. METHODS: Thirty-six patients with unilateral VS who underwent vestibular tests, including oVEMPs to BCV, cVEMPs to ACS, and caloric tests, were enrolled. The asymmetry ratios of the amplitudes of the oVEMPs to BCV and cVEMPs to ACS and canal paresis on the caloric test were measured. RESULTS: Among the 36 patients with VS, 31 (86%) showed reduced or absent oVEMPs to BCV. Twenty-eight patients (78%) showed abnormal cVEMPs to ACS, and 31 (86%) showed abnormal caloric responses. The consistency of the results between oVEMPs to BCV and the caloric test was significantly higher than that between oVEMPs to BCV and cVEMPs to ACS (p < 0.02). CONCLUSION: Ocular vestibular evoked myogenic potentials to bone-conducted vibration mainly reflect the function of the superior vestibular nerve. PMID- 19816225 TI - Cartilage plate tympanoplasty. AB - OBJECTIVES: The purpose of this work was to report our modified cartilage plate tympanoplasty technique ("tulip leaves") and to analyze its clinical outcome in primary and recurrent cases of chronic otitis media with and without cholesteatoma. STUDY DESIGN: Clinical retrospective study. METHODS: Patients being operated on with this technique at the University Department of Otorhinolaryngology, Dresden, Germany, between 1993 and 2001 were invited for survey, otomicroscopy, and pure-tone audiometry in 2003. Patients' charts were used to draw necessary conclusions. RESULTS: A total of 39 patients who were treated with this technique after canal wall down tympanomastoidectomy and cavity obliteration were included in this long-term analysis after a median follow-up of 6 years. Seventeen patients (44%) experienced chronic otitis media with cholesteatoma, whereas 22 (56%) of them had a diagnosis of chronic otitis media without cholesteatoma. At the time of examination, all patients displayed a closed tympanic membrane. However, retractions were observed in 19 patients (48%). One patient required (3%) revision surgery for recurrent cholesteatoma due to prosthesis extrusion during the study period. CONCLUSION: On the basis of this study, we recommend the tuliplike arrangement of thin but large auricular cartilage slices for the reconstruction of tympanic membrane defects in high-risk ears. This combination proved its high stability and long-lasting vitality in our long-term study. These characteristics are crucial for permanent disease removal and for reducing the risk of recurrent pathologic abnormality. PMID- 19816226 TI - Posterior semicircular canal dehiscence arising from temporal bone fibrous dysplasia. PMID- 19816227 TI - TNF-alpha receptor expression correlates with histologic activity of otosclerosis. AB - HYPOTHESIS: Otosclerosis is an inflammatory bone remodeling disorder of the human otic capsule, which might be characterized by variable levels and unique expression pattern of TNF-alpha receptors. BACKGROUND: Histologic characteristics of otosclerosis have been well described during the latest decades; however, the grading of different histopathologic and clinical stages has not been attributed precisely to the molecular biology of the pathologically increased metabolism of osteoclast-osteoblast axis. METHODS: Forty otosclerotic- and 40 nonotosclerotic ankylotic stapes footplates (n = 80; men, 29; women, 51) were histologically analyzed: conventional hematoxylin-eosin staining and tumor necrosis factor-alpha receptor I and II (TNFRI/II)-specific immunofluorescent assay was performed. RESULTS: Active otosclerosis (Grades I-II; n = 24) was featured by increased expression of TNFRII and moderate expression of TNFRI; inactive cases (Grades III IV) were characterized by permanent expression of TNFRI; however, TNFRII-specific immunoreaction was absent. Nonotosclerotic stapes specimens showed a negligible TNFR expression. Tumor necrosis factor receptor expression pattern showed a strong correlation with the histologic activity of otosclerosis (chi2 test; p < 0.001). CONCLUSION: Detection of elevated TNFR expression demonstrates activated osteoclast metabolism and inflammatory pathways in otosclerosis. Different etiopathogenesis of otosclerotic and nonotosclerotic stapes ankylosis should be distinguished. Administration of monoclonal anti-TNF-alpha antibody may be a reasonable option in the medical treatment of active stages of otosclerosis. PMID- 19816228 TI - Fisch's reversal steps stapedotomy: when to use it? AB - OBJECTIVE: To determine that the use of Fisch's reversal steps stapedotomy is recommended only when the visible portion of the footplate is blue in all its points before removing the stapes superstructure. STUDY DESIGN: Prospective study. MATERIALS AND METHODS: This study started on January 2007 and ended on June 2007, when the statistical data reached significance. Fifty patients with otosclerosis were examined. Four otosclerosis were excluded from the study following the exclusion criteria indicated by Fisch (obliterative otosclerosis and narrow oval niche). Finally, 46 otosclerosis patients were considered for the study. For all patients, stapedotomy procedures using the reversal steps technique by Fisch were planned. The study group was divided into 2 groups (A and B) on the basis of the footplate color before removing the stapes superstructure. Group A included otosclerosis with blue footplate (n = 34). Group B included otosclerosis with white footplate (n = 12). We estimated whether the reversal steps stapedotomy proposed by Fisch significantly avoids incus complications (luxation and subluxation) and stapes footplate complications (luxation and fracture) both in blue and white otosclerosis. MAIN OUTCOME MEASURES: Footplate color, incidence of complications. RESULTS: The footplate fenestration had not caused fractures or luxation of footplate in both groups. The fracture of the anterior crus had caused 5 footplate complications (2 luxations and 3 fractures) in Group B, but none in Group A. This difference was significant. In Group A, we had no subluxation/luxation of the incus. In Group B, we had 3 incus subluxations. This difference was significant. CONCLUSION: The use of Fisch's reversal steps stapedotomy is recommended only when the visible portion of the footplate, before removing the stapes superstructure, is blue in all its points, that is, "blue otosclerosis," because only in this case that the original idea of Fisch avoids incus and footplate complications. When the visible portion of the footplate, before removing the stapes superstructure, is white in all or in most of its points, that is, "white otosclerosis," the reversal steps technique by Fisch is not recommended because it does not avoid incus luxation/subluxation and footplate complications (fracture and luxation of the anterior half). PMID- 19816229 TI - Preliminary evaluation of a novel bone-conduction device for single-sided deafness. AB - HYPOTHESIS: A new intraoral bone-conduction device has advantages over existing bone-conduction devices for reducing the auditory deficits associated with single sided deafness (SSD). BACKGROUND: Existing bone-conduction devices effectively mitigate auditory deficits from single-sided deafness but have suboptimal microphone locations, limited frequency range, and/or require invasive surgery. A new device has been designed to improve microphone placement (in the ear canal of the deaf ear), provide a wider frequency range, and eliminate surgery by delivering bone-conduction signals to the teeth via a removable oral appliance. METHODS: Forces applied by the oral appliance were compared with forces typically experienced by the teeth from normal functions such as mastication or from other appliances. Tooth surface changes were measured on extracted teeth, and transducer temperature was measured under typical use conditions. Dynamic operating range, including gain, bandwidth, and maximum output limits, were determined from uncomfortable loudness levels and vibrotactile thresholds, and speech recognition scores were measured using normal-hearing subjects. Auditory performance in noise (Hearing in Noise Test) was measured in a limited sample of SSD subjects. Overall comfort, ease of insertion, and removal and visibility of the oral appliance in comparison with traditional hearing aids were measured using a rating scale. RESULTS: The oral appliance produces forces that are far below those experienced by the teeth from normal functions or conventional dental appliances. The bone-conduction signal level can be adjusted to prevent tactile perception yet provide sufficient gain and output at frequencies from 250 to 12,000 Hz. The device does not damage tooth surfaces nor produce heat, can be inserted and removed easily, and is as comfortable to wear as traditional hearing aids. The new microphone location has advantages for reducing the auditory deficits caused by SSD, including the potential to provide spatial cues introduced by reflections from the pinna, compared with microphone locations for existing devices. CONCLUSION: A new approach for SSD has been proposed that optimizes microphone location and delivers sound by bone conduction through a removable oral appliance. Measures in the laboratory using normal-hearing subjects indicate that the device provides useful gain and output for SSD patients, is comfortable, does not seem to have detrimental effects on oral function or oral health, and has several advantages over existing devices. Specifically, microphone placement is optimized for reducing the auditory deficit caused by SSD, frequency bandwidth is much greater, and the system does not require surgical placement. Auditory performance in a small sample of SSD subjects indicated a substantial advantage compared with not wearing the device. Future studies will involve performance measures on SSD patients wearing the device for longer periods. PMID- 19816230 TI - Deregulated genes in sporadic vestibular schwannomas. AB - OBJECTIVE: In search of genes associated with vestibular schwannoma tumorigenesis, this study examines the gene expression in human vestibular nerve versus vestibular schwannoma tissue samples using microarray technology. MATERIAL AND METHODS: RNA was extracted from 3 vestibular nerves (serving as control) and 16 solid, sporadic vestibular schwannomas. RNA (5 microg) was used in the labeling and biotinylation protocol to produce cRNA, which was hybridized to Affymetrix HG-U133A arrays. Data were imported into dChip v.1.3 and normalized using invariant set normalization. Differentially expressed genes were identified as differences between control and tumor tissue larger than 2-fold, with a conservative p value of less than 0.000161 and means of differences greater than 25. RESULTS: Eighty-seven probe sets, representing 78 genes, were significantly up- or down-regulated in tumor tissue. The deregulated genes were matched against established gene ontology, revealing that 8 of the up-regulated genes are involved in regulation of the cell cycle, 6 in cell morphogenesis, 8 in cell development, 11 in cell differentiation, 6 in cell death, 13 in cell adhesion, 9 in extracellular matrix, and 50 in protein binding (overlapping occurring). Gene annotation enrichment analyses of the clustered genes showed significant enrichment of annotations for the extracellular matrix (p < 0.0002), cell adhesion (p < 0.0001), and protein binding (p < 0.0004). CONCLUSION: We conclude that a number of transcripts are deregulated in sporadic vestibular schwannomas, and that several of these have functional annotations implicated in tumorigenesis. Specifically, genes involved in extracellular matrix function, cell adhesion, and protein binding seem to be of potential importance. However, further studies using other methodologies are needed for verification of the observed changes of gene expression seen by cDNA microarray analyses, for example, reverse-transcriptase-polymerase chain reaction and protein analyses. PMID- 19816231 TI - Hearing results using the SMart piston prosthesis. AB - OBJECTIVE: SMart, a newly introduced piston prosthesis for stapedotomy, is a nitinol-based, heat-activated, self-crimping prosthesis. We review our hearing results and postoperative complications using this self-crimped piston prosthesis and compare them with those obtained using stainless steel or platinum piston prostheses. HYPOTHESIS: Audiometric results using the SMart piston are identical to those obtained using a conventional piston prosthesis. STUDY DESIGN: Retrospective chart review. SETTING: Private neurotologic tertiary referral center. PATIENTS: The 416 ears reviewed included 306 with a SMart prosthesis and 110 conventional prostheses. 61% were women. Mean follow-up time was 5.6 (standard deviation [SD], 6.3 mo) and 6.9 months (SD, 7.0 mo) for the 2 groups, respectively. INTERVENTION: Stapedotomy using the SMart or a conventional (non SMart) prosthesis. MAIN OUTCOME MEASURES: Audiometric hearing results, including pure-tone average (PTA) and air-bone gap (ABG), and prevalence of postoperative complications. RESULTS: Mean postoperative PTA was 32.6 (SD, 16.8) dB for the SMart group and 29.4 (SD, 13.5) dB for the non-SMart group, with ABGs of 7.6 (SD, 8.9) and 6.0 (SD, 5.2) dB, respectively. Mean change (decrease) in ABG was 18.7 (SD, 13.1) dB for the SMart group and 19.9 (SD, 10.3) dB for the non-SMart group. High-frequency bone PTAs showed overclosure of 2.0 (SD, 7.9) dB for the SMart group and 3.6 (SD, 8.6) dB for the non-SMart group. Postoperative vertigo and tinnitus were infrequent. No significant differences in these audiometric outcomes or complication rates were noted between groups. There was no significant difference in rate of gap closure to within 10 dB (78.3 versus 84.2%, SMart and non-SMart, respectively) or 20 dB (94.2 and 98.0%). CONCLUSION: Compared with conventional stapes prostheses, the nitinol-based SMart is a safe and reliable stapes prosthesis that eliminates manual crimping without significantly altering the audiometric outcome. Complications are rare, but longer follow-up is needed before establishing long-term stability. PMID- 19816232 TI - Intraoperative electromyography and surgical observations as predictive factors of facial nerve outcome in vestibular schwannoma surgery. AB - OBJECTIVE: To evaluate the predictive value of electromyographic recordings for the immediate facial nerve outcome after solitary vestibular schwannoma (VS) removal. STUDY DESIGN: Prospective cohort study. SETTING: Tertiary referral center. PATIENTS AND METHODS: From November 2005 to July 2007, 120 operated solitary VSs monitored by NIM Response 2.0 (Medtronic, Xomed, Jacksonville, FL, USA) were included. Intraoperative direct facial nerve stimulation at the brainstem (proximal pre site) before resection and at the brainstem (proximal post and distal sites, respectively) and internal acoustic meatus after resection were performed. Intraoperative recordings were performed on 4 muscles: Frontalis, Orbicularis oculi, Orbicularis oris, and chin muscles (CM). Postoperative facial function (House-Brackmann classification) was evaluated at postoperative Days (D) 2, 8, and 30. Grades 1 or 2 were considered as a good result. RESULTS: A good postoperative facial function was obtained in 71, 63, and 77% at postoperative D2, D8, and D30, respectively. Maximal conduction block was found at Day 8. Poor facial nerve outcomes were observed for large tumor with a facial nerve superior to the tumor with a strong adhesion to the nerve. With a stimulation threshold at proximal site after proximal site tumor resection at less than or equal to 0.04 mA, a good facial function was observed in 77% at Day 8. Using these criteria together with CM responses greater than 800 muV to supramaximal stimulation at (proximal post site) and a proximal/distal post ratio at CM response greater than 0.6, 93% of patients presented with a good facial function at Day 8. CONCLUSION: Using CM recording stimulation and supramaximal stimulation of the facial nerve in combination with thresholds seem to increase the predictive value of the monitoring for the immediate facial nerve function for VS surgery. PMID- 19816233 TI - The impact of Type D personality on health-related quality of life in tinnitus patients is mainly mediated by anxiety and depression. AB - OBJECTIVE: To evaluate the impact of Type D personality on health-related quality of life (HRQoL) and self-reported tinnitus-related distress in chronic tinnitus patients and whether this relationship is mediated by indicators of psychological distress (i.e., vital exhaustion, anxiety, and depression). MATERIALS AND METHODS: Using a cross-sectional study design, 265 consecutive tinnitus patients were asked to complete the Hospital Anxiety and Depression Scale, the Maastricht Questionnaire, the Type D Scale (DS14), the Short-Form Health Survey 36, and the Tinnitus Reaction Questionnaire. RESULTS: The prevalence of Type D was 35.5%. Type D patients were significantly more anxious, depressed, and vitally exhausted, and experienced more impaired HRQoL and increased tinnitus-related distress compared with non-Type D patients. Structural equation modeling showed that Type D personality directly increased symptoms of depression and anxiety, but not vital exhaustion. Type D was also a direct predictor of poor mental and physical HRQoL and increased tinnitus-related distress, although this influence was mainly mediated by symptoms of depression and anxiety. Anxiety, depression, and vital exhaustion had a direct influence on HRQoL and self-reported tinnitus related distress, with a higher impact on mental HRQoL (R2 = 0.74) compared with physical HRQoL (R2 = 0.33). Vital exhaustion was a predictor of HRQoL and self reported tinnitus-related distress; however, its influence was moderated by enhanced levels of anxiety and depression. CONCLUSION: Tinnitus patients with a Type D personality were more likely to be anxious and depressed and to experience poor HRQoL and increased self-reported tinnitus-related distress, with the impact of Type D mainly being mediated by symptoms of anxiety and depression, although Type D also exerted a direct influence on these outcomes. These findings underline that to reduce the impact of tinnitus on HRQoL and self-reported tinnitus-related distress, treatment should be directed toward reducing anxiety and depression, especially in patients with a Type D personality. PMID- 19816234 TI - Eosinophilic inflammation in the middle ear induces deterioration of bone conduction hearing level in patients with eosinophilic otitis media. AB - OBJECTIVE: Eosinophilic otitis media (EOM) is characterized by the extensive accumulation of eosinophils in the middle ear mucosa and middle ear effusion and is usually associated with bronchial asthma. Eosinophilic otitis media patients show gradual or sudden deterioration of hearing. In our previous study, we reported that high-tone loss was more frequently found and more severe in EOM patients than in control patients with chronic otitis media. These findings suggest that not only bacterial infection but also eosinophilic inflammation in the middle ear may damage the inner ear. The present study was performed to determine whether eosinophilic inflammation is indeed related to deterioration of bone-conduction hearing level (BCHL). PATIENTS: Fifty-five ears of 28 patients with EOM associated with bronchial asthma were included in this study. Middle ear effusion (MEE) samples were collected from all the patients, and the concentrations of eosinophilic cationic protein (ECP) and immunoglobulin E (IgE) were measured by fluorescence enzyme immunoassay. The BCHLs at 2 and 4 kHz for the worse-hearing ear of each patient were correlated with the concentrations of ECP and IgE. RESULTS: The concentration of IgE in MEE significantly and positively correlated with BCHL at 2 and 4 kHz. The ears with a higher concentration of ECP in MEE also tended to show deterioration of BCHL at 4 kHz. Other clinical risk factors for BCHL deterioration were male sex, long duration of EOM, association with bacterial infection, severe inflammatory changes of the middle ear mucosa, and high serum IgE concentration. CONCLUSION: Eosinophilic inflammation-related substances such as ECP and IgE are closely related to the deterioration of BCHL at high frequencies. Particularly, IgE concentration in MEE is a good indicator of BCHL elevation. We should always pay attention to the hearing acuity of EOM patients with the risk factors. PMID- 19816235 TI - Intraventricular hemorrhage in premature infants: mechanism of disease. AB - Intraventricular hemorrhage (IVH) is a major complication of prematurity. IVH typically initiates in the germinal matrix, which is a richly vascularized collection of neuronal-glial precursor cells in the developing brain. The etiology of IVH is multifactorial and is primarily attributed to the intrinsic fragility of the germinal matrix vasculature and the disturbance in the cerebral blood flow (CBF). Although this review broadly describes the pathogenesis of IVH, the main focus is on the recent development in molecular mechanisms that elucidates the fragility of the germinal matrix vasculature. The microvasculature of the germinal matrix is frail because of an abundance of angiogenic blood vessels that exhibit paucity of pericytes, immaturity of basal lamina, and deficiency of glial fibrillary acidic protein (GFAP) in the ensheathing astrocytes endfeet. High VEGF and angiopoietin-2 levels activate a rapid angiogenesis in the germinal matrix. The elevation of these growth factors may be ascribed to a relative hypoxia of the germinal matrix perhaps resulting from high metabolic activity and oxygen consumption of the neural progenitor cells. Hence, the rapid stabilization of the angiogenic vessels and the restoration of normal CBF on the first day of life are potential strategies to prevent IVH in premature infants. PMID- 19816236 TI - IUGR does not predispose to necrotizing enterocolitis or compromise postnatal intestinal adaptation in preterm pigs. AB - IUGR and preterm birth are leading causes of neonatal morbidity. We tested the hypothesis that IUGR predisposes to gut maladaption and necrotizing enterocolitis (NEC) using preterm pigs as models for preterm infants. First, full-term normal birth weight (NW) and IUGR ( approximately 65% of NW) pigs were compared. IUGR reduced intestinal weight per length, proportion mucosa, villous area, and sucrase activity at 2 d after birth (p < 0.05) but did not change relative organ weights. Next, groups of preterm pigs were fed formula or colostrum, starting at birth or after 2-3 d of total parenteral nutrition (TPN). Neonatal mortality (not related to NEC) was increased in IUGR versus NW preterm pigs (28 vs 10%, p < 0.01). NEC incidence was similar between IUGR and NW but higher after formula than colostrum feeding (46 vs 12%, p < 0.01) and higher after TPN than without TPN (61 vs 34% for formula pigs, p < 0.01). After feeding, relative intestinal mass and length were higher in IUGR versus NW pigs (+25-80%, p < 0.05) while brush border enzyme activities were similar. An enhanced gut trophic response to enteral feeding may help to improve postnatal intestinal adaptation and NEC resistance in preterm IUGR newborns. ABBREVIATIONS: : PMID- 19816237 TI - VEGF polymorphisms are associated with endocardial cushion defects: a family based case-control study. AB - Endocardial cushion defects (ECDs) of the cardiac outflow tract are among the most common congenital heart disease phenotypes. VEGF is essential for endocardial cushion formation and derangements in VEGF synthesis lead to ECD. Three functional single nucleotide polymorphisms (SNPs) in the VEGF gene -2578 C>A, -1154 G>A, and -634 G>C play a role in cardiogenesis. In a Dutch case control family study of triads, 190 case and 317 control children with both parents, we investigated linkage and association between these VEGF SNPs and ECD. Allele frequencies for the three VEGF SNPs were comparable between ECD children and controls. However, VEGF alleles -2578 C and -1154 G were transmitted more frequently to children with ECD (p = 0.003 and p = 0.002), in particular perimembranous ventricular septal defects (p = 0.012 and p = 0.006). The -2578A/ 1154A/-634G haplotype was associated with a reduced risk of ECD (OR 0.7; 95% CI, 0.6-1.0) and was significantly less transmitted to children with ECD (p = 0.002). In a Dutch population, we show that the VEGF 2578 C, -1154 G alleles, and the AAG haplotype are associated with ECD. Possible VEGF gene-environment interactions exposures are discussed. ABBREVIATIONS: : PMID- 19816238 TI - Pre-S2 deletions of hepatitis B virus and hepatocellular carcinoma in children. AB - The cause of early oncogenesis in hepatitis B virus (HBV)-related childhood hepatocellular carcinoma (HCC) remains unclear. This study investigated whether pre-S deletion of HBV is related to childhood HCC. By using nested polymerase chain reaction, we compared the pre-S sequence of HBV from sera of children with HCC against control children with similar chronic HBV infection. The HBV in sera of children with HCC had a significantly higher rate of pre-S deletion than that of children with chronic HBV infection (p = 0.008). All except one of the pre-S deletions from the HCC group involved the pre-S2 region, whereas no pre-S2 deletion was found in the chronic HBV group (p = 0.003). There was a trend whereby genotype-C sera had a higher rate of pre-S2 deletion than genotype-B sera (p = 0.11). A multivariate logistic regression model revealed that pre-S deletion was an independent risk factor for HCC in children (odds ratio: 36.69, p = 0.015). In conclusion, pre-S2 deletion does not need to take decades to occur; its presence in nearly half of children with HCC, in contrast to its absence in children with chronic HBV infection, suggests a link between pre-S2 deletion and HCC development in children. ABBREVIATIONS: : PMID- 19816239 TI - Airway injury from initiating ventilation in preterm sheep. AB - Premature infants exposed to ventilation are at risk of developing bronchopulmonary dysplasia and persistent lung disease in childhood. We report where injury occurred within the lung after brief ventilation at birth. Preterm sheep (129 d gestation) were ventilated with an escalating tidal volume to 15 mL/kg by 15 min to injure the lungs, with the placental circulation intact (fetal) or after delivery (newborn). Fetal lambs were returned to the uterus for 2 h 45 min, whereas newborn lambs were maintained with gentle ventilatory support for the same period. The control group was not ventilated. Bronchoalveolar lavage fluid (BALF) and lung tissue were analyzed. In both fetal and newborn lambs, ventilation caused bronchial epithelial disruption in medium-sized airways. Early growth response protein 1 (Egr-1), monocyte chemotactic protein 1 (MCP-1), IL-6, and IL-1beta mRNA increased in the lung tissue from fetal and newborn lambs. Egr 1, MCP-1, and IL-6 mRNA were induced in mesenchymal cells surrounding small airways, whereas IL-1beta mRNA localized to the epithelium of medium/small airways. Ventilation caused loss of heat shock protein 70 (HSP70) mRNA from the bronchial epithelium, but induced mRNA in the smooth muscle surrounding large airways. HSP70 protein decreased in the lung tissue and increased in BALF with ventilation. Initiation of ventilation induced a stress response and inflammatory cytokines in small and medium-sized airways. PMID- 19816240 TI - Photodynamic effects on retinal oxygen saturation, blood flow, and electrophysiological function in patients with neovascular age-related macular degeneration. AB - PURPOSE: To evaluate the effect of photodynamic therapy (PDT) on retinal functions, such as oxygen saturation, blood flow, and electrophysiological function using Fourier transform-based spectral retinal imaging (SRI), Heidelberg retinal flowmeter (HRF), and multifocal electroretinogram (mfERG). METHODS: This was a prospective interventional case series. Twenty-two patients with age related macular degeneration (AMD) with unilateral choroidal neovascularization (CNV) were examined using SRI, HRF, and mfERG before and 1 week and 1 month after PDT. Eleven fellow eyes without CNV and 11 control eyes of 11 age-matched healthy volunteers were also investigated. Eleven of 22 patients with neovascular AMD were retreated using verteporfin PDT and examined using a protocol similar to the one used for the first treatment. RESULTS: Oxygen saturation levels in the macula of eyes with neovascular AMD were significantly higher than those in normal control eyes (P = 0.026) but were not significantly higher in eyes with nonneovascular AMD. Oxygen saturation levels decreased 1 week after a single treatment (P = 4.59 x 10(-3)) and retreatment (P = 0.0134) and recovered to baseline levels at 1 month follow-up (P > 0.05). HRF demonstrated reduced mean blood flow at 1 week after single treatment (P = 9.22 x 10(-4)) and retreatment (P = 0.0307). One month after PDT, mean blood flow tended to show recovery. There was a similar decrease in mfERG amplitude 1 week after treatment, but the logarithm of minimum angular resolution (logMAR) vision was stable or improved throughout follow-up. CONCLUSION: Oxygen saturation levels, mean blood flow, and mfERG amplitude decreased 1 week after PDT treatment in both single treatment and retreatment groups. Although logMAR vision is stable or improved, our data showed transient functional deterioration in the retina after PDT treatment. PMID- 19816241 TI - Postoperative complications and intraocular pressure in 943 consecutive cases of 23-gauge transconjunctival pars plana vitrectomy with 1-year follow-up. AB - BACKGROUND: To report the rate of postoperative complications in 943 consecutive eyes operated on with 23-gauge transconjunctival pars plana vitrectomy. METHODS: Single-center, retrospective, noncomparative, interventional case series. Nine hundred and forty-three eyes underwent 23-gauge transconjunctival core and peripheral vitrectomy with peripheral laser at the sclerotomy sites from May 2005 through April 2008. The main outcome measures were intraocular pressure at 1 day and at 1 week and intra- and postoperative complications with at least 6-month follow-up. RESULTS: Eight hundred and thirty-one eyes (88%) did not have either significant intra- or postoperative complications. Sclerotomy leakage requiring suture occurred in 37 eyes (3.9%). One choroidal detachment (0.1%) spontaneously resolved 1 week after surgery. At postoperative Day 1, 31 eyes (3.3%) experienced transient hypotony. Forty-five eyes (4.8%) presented a subtle vitreous hemorrhage that resolved spontaneously. Two retinal detachments (0.2%) occurred, one at 1 month and one at 3 months. They resolved with one further vitrectomy. Seven hundred and forty eyes completed the 12-month follow-up and presented no further complications. CONCLUSION: Twenty-three-gauge complete vitrectomy and peripheral laser seem safe for a variety of vitreoretinal surgical procedures. The rate of post- and intraoperative complications compares favorably with 25-gauge and with the standard 20-gauge vitrectomy. PMID- 19816242 TI - A prospective study of blood pressure and intraocular pressure changes in hypertensive and nonhypertensive patients after intravitreal bevacizumab injection. AB - PURPOSE: To evaluate the changes in blood pressure and intraocular pressure (IOP) in patients with and without hypertension after intravitreal bevacizumab (Avastin) injection. METHODS: This study examined 135 (74 nonhypertensive vs. 61 hypertensive) consecutive patients treated with an intravitreal injection of 1.25 mg bevacizumab for retinal vascular diseases prospectively over a 6-month period. All the hypertensive patients were on medication, and only patients with controlled hypertension after consultation with cardiology specialists were included. Intraocular pressure and blood pressure were measured before and 30 minutes, 1 day, 1 week, 3 weeks, and then every month after the bevacizumab injection. RESULTS: The mean baseline systolic and diastolic pressures were significantly higher in the hypertensive group (systolic P = 0.001, diastolic P = 0.023). None of the systolic and diastolic values after injection differed significantly from the baseline values in the hypertensive group. By contrast, the 30-minute systolic values were significantly higher than baseline (P < 0.05), and the 1-day diastolic values were lower than before injection in the nonhypertensive group (P < 0.05). The mean baseline IOP of the nonhypertensive and hypertensive groups was similar (P = 0.08). The mean IOP 30 minutes after injection was significantly higher than the baseline mean IOP (P < 0.05), and the mean IOPs at 1 day and 1 week were significantly lower than before injection in both groups (P < 0.05). CONCLUSION: Intravitreal bevacizumab injection is safe in terms of blood pressure and IOP in both hypertensive and nonhypertensive patients after 6 months of follow-up. PMID- 19816243 TI - State of the art management in spine oncology: a worldwide perspective on its evolution, current state, and future. AB - STUDY DESIGN: A review of the past and current status of the evolving field of spine oncology. OBJECTIVE: To provide a framework of reference for developments in the field, particularly the rapidly evolving field of molecular biology, as well as contemporary practice in the management of spine tumors. METHODS: Literature review of the surgical treatment of spine tumors in the past and present, the emerging radiologic and biologic technologies, as well as the field of targeted therapy in cancer and the economic implications of technological advances. RESULTS: A vast contemporary literature is currently available that provides a clear rational basis for treatment. Most treatment recommendations are currently based on retrospective data and small Phase II prospective studies. Treatment paradigms continue to evolve without their relative merits being evaluated by randomized controlled trials. The current lack of randomized trials in spine oncology reflect both the rarity of spine tumors and strongly held biases based on retrospective studies and institutional bias. CONCLUSION: Spine oncology is a rapidly evolving field with contributions in surgery, radiation therapy, and targeted chemotherapy resulting in overall improvement in quality of life and survival in patients with spine tumors. However, the economic consequences of these improvements are substantial and need to be kept in proper perspective. PMID- 19816244 TI - Spine focus issue. Summary of management recommendations in spine oncology. PMID- 19816245 TI - Innovations in cleft surgery. PMID- 19816246 TI - Communication skills for surgeons and their teams: publishing medical research. PMID- 19816247 TI - Back to the future. PMID- 19816248 TI - Genetic aspects of cognition and speech production: going beyond surgical repair. PMID- 19816249 TI - Tbx22 expressions during palatal development in fetuses with glucocorticoid /alcohol-induced C57BL/6N cleft palates. AB - T-box transcription factor 22 (Tbx22) belongs to the T-box family of transcription factors and was originally found using an in silico approach to identify new genes in the human Xq12-Xq21 region. Mutations in Tbx22 have been reported in families with X-linked cleft palate and ankyloglossia, but the underlying pathogenetic mechanism remains unknown. The aim of this study was to evaluate the expression of Tbx22 messenger RNA (mRNA) during palatogenesis in glucocorticoid-/alcohol-induced cleft palate in a C57BL/6N mouse model. Palatal development was monitored by histomorphologic and immunohistochemical studies and by in situ hybridization. Thirty pregnant C57BL/6N mice at 8 weeks of age, weighing 20 to 25 g, were used in this study. In the experimental group, 12 mice were exposed to alcohol for 7 days before mating, and 12 mice in the control group were not exposed. Six mice in a sham group were exposed to neither alcohol nor glucocorticoids. A total of 18 fetuses with induced cleft palates each from 102 fetuses in the experimental group, 109 in the control group, and 58 in the sham group were used. In both the experimental and the control groups, glucocorticoids were injected subcutaneously on gestational days (GD) 9.5, 10.5, and 11.5, and each mouse was killed on GDs 10.5 to 15.5. Histomorphologic findings were studied using hematoxylin and eosin staining, and antibodies against proliferation cell nuclear antigen, matrix metallopeptidase 9, zinc finger protein 422 (Krox25) heat shock protein 70, and Tbx22 were used in immunohistochemical analysis. Mouse Tbx22 mRNA was identified, and its expression was analyzed during embryogenesis by polymerase chain reaction and in situ hybridization. Coronal sections of the cleft maxilla of the embryos with induced cleft palates had a gap between the palatal shelves, where 2 palatal shelves had fused as in normal development but failed to meet and fuse to each other. By in situ hybridization, Tbx22 mRNA was found to be expressed in distinct areas of the head, such as the mesenchyme of the inferior nasal septum, the posterior palatal shelf before fusion, and the attachment of the tongue during normal development of the palate and maxilla from GD 11.5. Localization in the tongue frenum correlated with the ankyloglossia phenotype in the induced cleft palate animal model. PMID- 19816250 TI - Nasal tip complications of primary cleft lip nasoplasty. AB - BACKGROUND AND PURPOSE: Complications of primary nasoplasty, at the time of definitive primary cheiloplasty, are underreported in the literature. This study endeavors to examine the occurrences of these complications at our cleft craniofacial center, in an effort to identify causative factors and management strategies. A case series of patients with postoperative nasal complications after primary cleft lip nasal surgery is presented. METHODS: A retrospective chart review of primary cleft lip nasal repairs was conducted at our cleft craniofacial center between January 2003 and December 2007. Consecutive cases of 3 staff surgeons were evaluated. Specific data points included number and type of complications, subsequent required interventions, and relevant history, with particular attention paid to the details of the primary nasoplasty. RESULTS: Eighty-six primary cleft lip nasoplasties were completed between the years 2003 and 2007. Six complications (6.9%) related to the primary cleft lip nasoplasty were identified. Four patients (4.6%) experienced nasal tip infections; all 4 required surgical drainage. Twenty-four patients (27.9%) undergoing primary cleft lip and nose repair had postoperative nostril conformers placed, and 2 (8.3%) of them experienced complications deemed conformer related. CONCLUSIONS: Postoperative nasal complications of primary cheiloplasty occur and are likely underreported. In this series, complications resulted from infection, often occurring late, and secondary to the use of nostril conformers. Surgeon awareness and caregiver education, to identify the early signs of postoperative nasal complications, are critical to the successful treatment of these occurrences. Although this study did not intend on examining antibiotic use, the significance of nasal tip infections might support the regular use of antibiotics in this population, and the use of postoperative nostril conformers must be followed closely. PMID- 19816251 TI - Hypopharyngeal reconstruction using remnant narrow pharyngeal wall as omega shaped radial forearm free flap. AB - PURPOSE: There have been numerous attempts to use pharyngoesophageal reconstruction to restore swallowing function. Much controversy exist over reconstruction after total or partial pharyngectomy, and there is also debate over whether to continue with the reconstructive procedure with a narrow strip of remnant mucosa or complete it after total pharyngectomy. We analyzed the utility of omega-shaped radial forearm free flap (RFFF) using the narrow remnant posterior pharyngeal wall. METHODS: Patients in group 1 (n = 12) had a narrow remnant pharyngeal wall with a width of less than 3 cm. Those in group 2 (n = 35) had a remnant pharynx with a width larger than 3 cm. The incidence of fistula, stricture, and swallowing difficulty were evaluated. Swallowing difficulty was graded using a 7-point visual analog scale. All circumferential hypopharyngeal reconstruction with tubed RFFF, pectoralis major flap, and jejunal free flap were also compared with group 1. RESULTS: All flaps survived, and 1 fistula (8%) was detected in group 1. Compared with tubed RFFF (46%) and tubed pectoralis major flap (57%), this is a relatively low rate of fistula formation. In group 1, normal diet was possible in 92% of patients, but 1 patient can tolerate a liquid diet only. In group 2, normal diet was possible in 80% of patients. As for swallowing difficulty, the median visual analog scale score for both groups 1 and 2 was 6 points. When comparing different flaps, stricture and fistula rate was 0 and 8% in group 1, 15 and 46% in tubed RFFF, 43 and 57% in tubed pectoralis major flap, and 33 and 5% in jejunal flap, respectively. CONCLUSIONS: We performed all surgeries taking care not to transgress the wide excision principle with the remnant hypopharyngeal wall. For remnant lesions greater than 3 cm, patch-type RFFF was performed, whereas for those less than 3 cm, omega-shaped RFFF was done. We achieved fairly good results in both groups without total resection of the narrow remnant hypopharyngeal wall. PMID- 19816252 TI - Airway analysis: with bilateral distraction of the infant mandible. AB - BACKGROUND: Mandibular distraction was proven to be a valuable tool for lengthening the hypoplastic mandible and relieving airway obstruction in infants. However, analysis of presurgical and postsurgical three-dimensional computed tomography and polysomnogram studies is lacking. The aim of this study was to describe the effect of distraction on the airway by evaluating the clinical, three-dimensional radiographic and polysomnogram studies before and after distraction. METHODS: Seventeen infants with micrognathia who underwent internal curvilinear mandibular distraction from April 2005 through April 2008 at Lucile Packard Children's Hospital were included. Preoperative and postoperative computed tomography, polysomnograms, and feeding evaluations were obtained and compared after distraction. RESULTS: The mean patient age before surgery was 105 days. All patients tolerated the distraction process with a mean mandibular advancement of 18.1 mm. One patient experienced a temporary marginal mandibular nerve palsy that resolved, and 1 postoperative wound infection was encountered. Preoperatively, the mean retroglossal oropharyngeal cross-sectional area was 41.53 mm. This was associated with a mean preoperative apnea-hypopnea index (AHI) of 10.57 and a minimum oxygen desaturation of 83%. After distraction, the mean airway increased to 127.77 mm. All patients had clinical improvement of their respiratory status; the mean postoperative AHI was 2.21, and the minimum oxygen desaturation was 90%. The result was a 209% cross-sectional airway increase. All patients progressed to oral feeds by 3.5 months postoperatively. CONCLUSIONS: Mandibular distraction is effective at relieving anatomic airway obstruction in infants with micrognathia and obstructive sleep apnea while avoiding some previously reported associated complications. PMID- 19816253 TI - Anthropometric measurement and analysis of the external nasal soft tissue in 119 young Han Chinese adults. AB - A random sample of 119 young, healthy Han Chinese adults (56 men and 63 women) between the age of 18 and 25 years (mean, 22.7 y) in PR China was obtained for this study. By the guidance of standard methods, based on Farkas's anthropometric measurements in craniofacial region, 12 nasal soft tissue landmarks and 12 linear and 3 angular measurements were chosen. The linear measurements were taken directly, whereas the angular measurements were taken by photogrammetric method. Eight nasal proportion indices were calculated according to the linear measurements. The application of the independent-samples t-test showed sex dimorphism in most parameters of the nasal region. All the linear measurements were larger in men than in women, whereas all the angular measurements were smaller in men than in women. The significant differences in partial parameters between men and women have been proved. Ten of 12 linear measurements, 1 of 3 angular measurements, and 3 of 8 nasal proportion indices showed significant sexual dimorphism (P < 0.01). Compared with other racial/ethnic groups, the nasal anthropometric measurements and proportion indices of Han Chinese adults were different, to some extent. This study could provide credible and objective reference material for plastic and maxillofacial surgeons for the external nasal soft tissue evaluation and planning of the cosmetic nasal surgery. Besides, these results could be a useful guidance for preoperative and postoperative evaluations of secondary rhinoplasty in nasal deformity associated with cleft lip and palate. PMID- 19816254 TI - Cleft lip and palate: a descriptive comparative, retrospective, and prospective study of patients with cleft deformities managed at 2 hospitals in Kenya. AB - This was a combined retrospective and prospective study in which 2 sets of results from 2 hospitals in Nairobi were analyzed and compared. The retrospective study was conducted at Kenyatta National Hospital, whereas the prospective study was conducted at Metropolitan Hospital.The main objective of the study was to establish the presentation and pattern of patients with cleft lip and palate and complications of repair at the 2 hospitals.In the retrospective arm of the study, files of all patients presenting with clefts at Kenyatta National Hospital between January 1998 and December 2007 were retrieved, and a questionnaire was filled out for each of them, whereas all patients seen and operated on for clefts at the Metropolitan Hospital from January 2007 to October 2008 were recruited into the prospective study.There was a predominance of male participants in both studies, and most clefts were on the left side. The retrospective and prospective studies had positive family history in 3.5% and 30.9%, respectively. Associated congenital malformations were 8.2% for the retrospective study and 25% for the prospective study. In both studies, the central province had the largest number of clefts, whereas the coast province had very few.Cleft lip and palate is a significant congenital malformation in Kenya, and there seems to be a higher incidence of familial tendency and associated congenital malformations than that reported elsewhere. PMID- 19816255 TI - Open approach via the nasal dorsum with dermal graft for bifid nose. AB - Bifid nose is a rare congenital anomaly and is also called double or cleft nose. The mode of appearance varies: from a simple groove at the nasal apex to a maxillary cleft. Neither surgical procedures nor the best time of surgery has been established. In this study, we performed dermal graft using an open approach via the nasal dorsum in patients with bifid nose and obtained good results. In the operation, a surplus skin of the nasal dorsum was resected through the nasal dorsum as a spindle graft and stored. The resected skin was denuded and the dermal graft applied on the nasal apex. For external nose plasty, it remains controversial which of 2 procedures, open or closed approach, should be selected. We have used the open approach to obtain a good surgical field. Nasal cartilage displacement could be confirmed under direct vision, facilitating cartilage correction. Furthermore, we could use the surplus skin component that is routinely abandoned and achieve a favorable nasal apex shape without adding supplemental invasiveness. PMID- 19816256 TI - Course and distribution of the lingual nerve in the ventral tongue region: anatomical considerations for frenectomy. AB - The aim of this study was to elucidate the sublingual and intralingual courses of the lingual nerve (LN) in the ventral tongue region, providing a clinical guide for safe surgical procedures such as frenectomy. We evaluated 16 specimens (32 sides) by gross observation after detailed dissections, and a further 6 specimens were examined after Sihler staining. All specimens were harvested from embalmed Korean cadavers. We classified the innervation patterns of the LN into 5 types and confirmed the distribution of the LN in the tip of the tongue. The classification of the LN was made with reference to a line formed by the interlacing of the styloglossus and genioglossus muscles. Based on the course of LN and the presence of a tiny twig (twigs directly innervating the ventral mucosa of the tongue, TM) directly innervating the sublingual mucosa, the course of the LN was classified as being straight, curved, or vertical and with or without the TM. Straight, curved, and vertical courses without the TM were seen in 9.4%, 46.9%, and 18.8% of the cases, respectively. Straight and curved courses with the TM were observed in 6.3% and 18.8% of the cases, respectively. Sihler staining revealed that the tongue tip is innervated by the LN. These findings indicate that surgical manipulations at the ventral tongue region might damage the LN and result in numbness of the tongue tip, and provide a useful anatomic reference for various surgical procedures involving the ventral tongue region. PMID- 19816257 TI - Volumetric assessment of lateral pterygoid muscle in unilateral chewing: a stereologic study. AB - Unilateral chewing habit is common in dentate populations. It may initiate a hypertrophy process in the lateral pterygoid muscles compared with the nonused sides. We hypothesized that the lateral pterygoid muscles of the affected sides would be larger in volume than those of the nonaffected sides in subjects with unilateral chewing habit. Because analysis of medical images in accordance with the rules of modern stereology enables precise unbiased estimates of muscle volume, and magnetic resonance imaging (MRI) is a criterion standard as an imaging modality for the analysis of muscles because of its excellent soft-tissue resolution, we estimated lateral pterygoid muscle volume both at right and left sides using stereologic techniques (Cavalieri principle) by means of MRI. The MRI estimated volumes of both the inferior and the superior heads of the lateral pterygoid muscle at the chewing side were significantly greater than the opposite side when compared. This finding may assist in the interpretation of lateral pterygoid muscle and temporomandibular joint-related problems in maxillofacial surgery clinics. This investigation has also demonstrated that systematically sampled MRI slices through the lateral pterygoid muscle can be used to obtain unbiased estimates. PMID- 19816258 TI - Modified design of Cupid's bow in the repair of unilateral microform cleft lip: in case of deficient distance between the midline and the cleft side Cupid's bow peak. AB - In unilateral microform cleft lip repair, when the distance from the cleft side peak of Cupid's bow to the midline is shorter than on the noncleft side, conventional cheiloplasty design results in asymmetric Cupid's bow with unnatural philtrum that is hard to correct. We modified the design of the cleft side Cupid's bow to avoid causing this secondary deformity. Twelve patients with a unilateral microform cleft lip who had a shorter cleft side from the midline to the peak of Cupid's bow distance were treated using a modified cheiloplasty design between January 1998 and December 2007. The point for the peak of Cupid's bow of the lateral segment was shifted medially to compensate for the short medial segment. The mean age of patients at the time of surgery was 48 months (range, 3 mo to 24 yr), and the follow-up period ranged from 10 to 100 months, with a mean of 33 months. A symmetric Cupid's bow with a natural-looking philtrum was achieved and maintained throughout the follow-up period in all the patients. In patients with a unilateral microform cleft lip with a shorter cleft side from the midline to the peak of Cupid's bow distance, the point for the peak of Cupid's bow of the lateral segment was shifted medially to achieve symmetric Cupid's bow. The resulting scar was more medial than in a conventional design, but the philtrum looked more natural. Longer follow-up would be required to verify the long-term results. PMID- 19816259 TI - Consideration of median cleft lip with frenulum labii superior. AB - Median cleft lip is usually divided into true and false when being discussed. Owing to recent developments of diagnostic imaging methods that have improved the accuracy, the presence of an intermediate type of median cleft lip, which cannot simply be divided into true and false, has been suggested. However, the simple method of classification is still clinically valuable. We have previously reported in this Journal a case of median cleft lip with 2 upper labial frenums. In the present study, based on our experience with false median cleft lip, we set forth a hypothesis that 2 upper labial frenums can be found in true median cleft lip, whereas no upper labial frenum is found in false median cleft lip. A review of the results of previous Japanese cases (7 true and 4 false cases) supported our hypothesis. We also reviewed one of our cases of right cleft lip accompanied by holoprosencephaly and discuss the case from the developmental perspective. The shape of the upper labial frenum may be a factor that can be used for clinically classifying intermediate median cleft lip into either true or false in cases that are otherwise difficult to classify. PMID- 19816260 TI - Cleft palate in Pfeiffer syndrome. AB - The frequency of associated cleft palate is known to be high in some fibroblast growth factor receptor 2 (FGFR2)-mediated craniosynostosis syndromes, such as Apert syndrome. However, there is little information on the frequency of palatal clefts in the FGFR2-mediated disorder, that is, Pfeiffer syndrome. The purpose of this study was to determine the frequency of palatal clefts in patients with Pfeiffer syndrome. The records of patients with Pfeiffer syndrome managed in our craniofacial unit were reviewed. Only patients with a confirmed diagnosis of Pfeiffer syndrome were included. Diagnostic criteria were as follows: characteristic mutations in FGFR1 or FGFR2 or, in the absence of genetic testing, clinical findings consistent with Pfeiffer syndrome as determined by a clinical geneticist or our most experienced surgeon (J.B.M.). Only 2 clefts were noted in 25 patients (8%), including 1 with a submucous cleft and 1 with an overt palatal cleft. Many patients (87%) were described as having a high-arched and narrow palate, and 1 had a low, broad palate. Nine patients were noted to have choanal atresia or stenosis. Clefting of the palate does occur in Pfeiffer syndrome but at a low frequency. PMID- 19816261 TI - Cumulative frequency distribution in East Asian facial widths using the facial golden mask. AB - INTRODUCTION: Many studies have been published on the clinical importance and attractiveness of a face. On the other hand, Asian faces give an impression that is not polished owing to their pronounced cheekbone and mandible. There have already been many studies in progress pertaining to measurements and analyses of Asian facial structure; however, the authors would like to contribute to such research by providing practical analytical data and distributional patterns of the widths of their faces showing how wide their faces are. MATERIALS AND METHODS: The facial golden mask was applied to 1000 facial photographs of Korean college students, of whom 500 were males and 500 were females.The horizontal ratios (HR) of the mid- and lower faces of each student were calculated against the facial golden mask. The results were divided into 3 groups according the degrees of the HR for further analyses: group 1 (HR = 1.0), group 2 (HR > 1.0), and group 3 (HR < 1.0). RESULTS: We obtained an S-shaped cumulative frequency distribution graph for young Koreans per sex, per midface, and per lower face. The graph is dense at the center and is shifted to the right, signifying that the Koreans' midfaces and lower faces are wider than that of the facial golden mask. CONCLUSIONS: Conclusion reached in this study is that the faces of Koreans are wider than those of the facial golden mask as commonly perceived; thus, it is expected that graphs for the cumulative frequency of the distribution will be used extensively in the future. PMID- 19816262 TI - Cleft lip and palate: current status from the literature and our experience. AB - Many years after surgical correction, a complete unilateral or bilateral cleft is inclined to show an inaesthetism often associated with functional defects. This sequela disturbs the facial growth during childhood. Across the world, each surgical school uses its own protocol, but which is the best surgical protocol for patients with cleft? The aim of this study was to present a review of international literature concerning surgical techniques for the repair of cleft lip and palate (CLP) in children and to report our personal surgical techniques in this field. We focus on the main role of the primary surgery and propose a personalized protocol therapy, depending on the severity of the cleft. On 36 patients, most of them showed unilateral CLP at birth; only 4 showed bilateral cleft. In this study, we used 36 patients without cleft but with class I occlusion for comparison purposes. Analysis of the 2 groups regarding the development of the maxillary arch and the evaluation of palatal morphology was carried out using lateral cephalograms and dental casts. The main result showed 28 patients with acceptable teeth occlusion and speech quality, a valid nasal function, and a proper aesthetic aspect. Controversy still exists regarding the optimum timing and surgical technique for CLP repair. We propose the creation of a scientific database on internationally recognized protocol as a starting point depending on the severity of the cleft, thus avoiding controversies in CLP therapeutic treatment. PMID- 19816263 TI - Surgical management of craniofacial and skull base rhabdomyosarcomas. AB - INTRODUCTION: Craniofacial and skull base rhabdomyosarcomas (RMSs) are rare. Surgeons are involved both in the resection and the reconstruction of these tumors and in the management of late sequelae of earlier treatment. These tumors are highly heterogeneous, and the initial presentation may be insidious. We wished to determine how diagnostic difficulties could be overcome and how management outcomes could be optimized. MATERIALS AND METHODS: All cases of RMS in patients who presented to our craniofacial program between 2004 and 2008 were retrospectively reviewed. RESULTS: Eight consecutive patients with RMS were treated. Six patients are Maori. Four patients had surgical resections of their primary tumors including 3 with curative intent. One had a recurrent tumor, and 3 had functional reconstruction after an earlier treatment. Inaccuracies in histologic diagnosis occurred in 2 patients. Two patients developed postoperative cerebrospinal fluid leakage. Two patients had leptomeningeal involvement and died within 10 months. All patients treated with curative intent achieved complete macroscopic margins. Two remained disease-free at 10 and 22 months. DISCUSSION: Treatment must be in the context of a multidisciplinary team with the appropriate craniofacial, neurosurgical, and reconstructive skills. Expedient and accurate histologic diagnoses by experienced pathologists are critical in optimizing the outcomes. Oncologists must be well resourced to administer chemoradiation in a timely manner. Leptomeningeal invasion is a dire prognostic feature. Cerebrospinal fluid leak after resection must be identified early and repaired. Late consequences of early childhood craniofacial cancer treatment may be ameliorated by tailored reconstructions. The predominance of Maori in our series has not been reported previously. PMID- 19816264 TI - Surgical repair of clefts: a system of operations and maintenance programs. PMID- 19816265 TI - Evaluation of a single miniplate use in treatment of zygomatic bone fracture. AB - Different methods of internal fixation of simple displaced zygomatic fracture, such as wiring, miniplate, and screw fixation, were compared for postreduction rotational stability caused by muscular forces. This study was performed to evaluate the minimally invasive therapy in cases of zygomatic bone fracture. Seventeen patients were treated by percutaneous hook reduction and miniplate fixation along the frontozygomatic suture. Postoperatively, repositioning of aesthetic and stability and also bone ends approximation were assessed clinically and radiologically. The patients were followed up for 6 to 49 months. Preoperative symptoms were subsided except the infraorbital sensitivity disturbances in one of the patients. Postoperative complications such as diplopia and hematoma were minimal and subsided by time. In this study, orbitozygomatic, commuted, and zygomatic bone fractures simultaneously with diplopia were excluded. We showed that treatment of an isolated zygomatic bone fracture according to aesthetic and functional requirements may be achieved by insertion of a single miniplate at the lateral rim of the orbit. PMID- 19816266 TI - Endoscopic removal technique of a huge ethmoido-orbital osteoma. AB - Osteomas are slow-growing benign tumors of the paranasal sinuses. They originate from the sinus wall and generally fills the sinus cavity. Many osteomas are asymptomatic and diagnosed incidentally on radiographs. The well-circumscribed, dense bony appearance on radiographs is usually diagnostic. Osteomas become symptomatic when they extend to the orbit or cranium. We report a rare case of ethmoido-orbital osteoma. Case reports and a review of the literature concerning osteoma and surgical techniques are presented. Treatment is not recommended in asymptomatic osteomas. If treatment is indicated, external or endoscopic approach can be chosen. The choice of surgical approach depends on the size of the lesion, location, and the experience of the surgeon. PMID- 19816267 TI - Incidence of otitis media in children with deformational plagiocephaly. AB - In the past decade, deformational plagiocephaly has seen a staggering increase attributed to the Back to Sleep Campaign of April 1992. With this increase, the possible clinical associations need to be fully understood. The anatomic changes seen include ipsilateral occipital flattening, ipsilateral frontal bossing, ipsilateral anterior ear displacement, contralateral occipital bossing, and contralateral frontal flattening [J Craniofac Surg 2004;15:368-372, erratum in J Craniofac Surg 2004;15:705; Fig. l]. Children with deformational plagiocephaly can have malpositioned ears affecting normal drainage of the eustachian tube as it shifts with this deformity. The eustachian tube plays an important role in draining the middle ear. As evidenced by children with cleft palates, structural differences in the anatomy of the middle ear and eustachian tube can result in an increased susceptibility to otitis media. An institutional review board-approved retrospective study was conducted on all patients whose conditions were diagnosed as deformational plagiocephaly from a cleft and craniofacial deformities clinic for a 2-year period. Parents of patients self-reported the number of ear infections on a questionnaire. The questionnaire results were compared to age matched patient data available from the Center for Disease Control. There were 1259 patients who fit the inclusion criteria for the questionnaire part of the study, and of these, 634 (50.4%) reported at least 1 ear infection by 1 year of age. In addition to the questionnaire, a subset of infants with deformational plagiocephaly up to 1 year of age were administered a tympanogram, to assess the state of the middle ear. There were 124 patients who fit the inclusion criteria for the tympanometry part of the study, and of these 121 had an abnormal tympanogram at 1 or more clinic visits. The questionnaire did not show deformational plagiocephaly to be a significant risk factor for otitis media in our patient population; however, a trend directly correlating otitis media and the severity level of deformational plagiocephaly was observed. Although this trend lacked statistical significance, further studies should explore these differences. Tympanogrametry showed a marked percentage of infants with deformational plagiocephaly to have eustachian tube dysfunction. Future studies will focus on the significance of these findings. PMID- 19816268 TI - Acquired entropion associated with the transconjunctival incision for facial fracture management. AB - Transconjunctival incisions are used in cases of orbitozygomatic trauma and elective lower eyelid blepharoplasty. Although this approach has reduced the rates of ectropion, there is a rare but reported incidence of entropion. We review our experience with craniofacial trauma between 1998 and 2008 and our use of the transconjunctival, subtarsal, and subciliary approaches. We describe 2 patients where entropion occurred after a transconjunctival approach for repair of orbital and zygomatic fractures and review the literature. With the increasing popularity of this incision in reconstructive and cosmetic plastic surgery, a complete understanding of orbital anatomy and associated treatment options for entropion is necessary. PMID- 19816269 TI - Transconjunctival approach to the orbit. PMID- 19816270 TI - Curvilinear mandibular distraction in a patient with mandibulofacial dysostosis associated with Diamond-Blackfan anemia. AB - Diamond-Blackfan anemia (DBA) is a congenital erythroid aplasia or hypoplasia presenting in infancy as a macrocytic anemia. It has been linked to a gene defect resulting in the absence of specific ribosomal proteins that enable erythroid maturation. Treacher Collins syndrome is also associated with defective ribosomal biogenesis. Fifty percent of patients with DBA also present with additional physical abnormalities including growth delay, craniofacial abnormalities, upper limp abnormalities, cardiac defects, urogenital malformations, and cleft palate. Upper airway obstruction may be a complication from craniofacial disorders and may be responsible for life-threatening sleep apnea that may result in pneumonia or respiratory failure. This report is of a patient with DBA who presented at birth with respiratory distress secondary to tongue-based obstruction of the airway due to mandibulofacial dysostosis leading to need for intubation. In an effort to avoid a tracheostomy, the patient underwent bilateral internal mandibular osteodistraction on day 4 of life with a latency period of 0 days and a distraction at a rate of 2 mm/d for 10 days until a class 3 dental occlusion existed with 5 mm of overcorrection. In addition, the patient was born with choanal atresia that was repaired at day 30 of life. After completion of the mandibular distraction, the patient has avoided tracheostomy and is now 6 months of age. Recent polysomnogram has demonstrated no significant apnea or hypopnea. PMID- 19816271 TI - Le Fort I osteotomy to correct malocclusion after reconstruction of the maxilla with the free fibula flap. AB - Malocclusion and facial asymmetry may follow microsurgical jawbone reconstruction. We describe the use of a Le Fort I osteotomy to correct malocclusion after fibula flap reconstruction of the maxilla. A 49-year-old patient with an extremely atrophied maxilla underwent alveolar crest augmentation by free fibula transfer. Bone healing was uneventful, but gross asymmetry of the reconstructed maxilla was apparent 3 months after surgery, with canting of the alveolar bone on the right side and residual skeletal discrepancy in the sagittal plane. A Le Fort I osteotomy was planned to correct malocclusion 6 months after fibula transfer. The maxilla was moved downward and forward and impacted in the right molar region. There were no postoperative complications. Solid bone union was achieved between the mobilized maxilla and the buttresses 3 months after surgery. At that time, osteointegrated implants were inserted, and an implant supported prosthesis was completed. Neither bone resorption nor implant failure was encountered after 12 months of masticatory loading. Surgical correction of malocclusion after maxillary bone augmentation with the fibula flap is possible. Le Fort I osteotomy represents a reasonable option after microvascular alveolar bone reconstruction of the maxilla, when additional movements are required to restore facial symmetry and occlusion. PMID- 19816272 TI - Evaluation of effects on bone tissue of different osteotomy techniques. AB - The aim of this study is to evaluate and compare the response of bone tissue after osteotomy carried out with either rotating cutters or with piezoelectric terminals.Bioptic samples of bone tissue were taken during operations with rotating burs and piezoelectric terminals to increase bone volume before implantology. Samples first underwent histomorphometric analysis. Subsequently, osteoblastic cells, obtained from different samples, were placed in culture and allowed to proliferate to in vitro evaluate the time to initiate growth and to reach confluence. Finally, a molecular biologic study by reverse transcription polymerase chain reaction was performed to evaluate the expression of typical osteoblastic molecular markers, such as osteoprotegerin and osteopontin.Histomorphometric analysis showed that the width of necrotic line on the osteotomic margins from samples taken using different techniques did not vary significantly. Moreover, the times of initial growth and of confluence in cells from the 2 groups did not show any statistically significant differences. However, a highly significant correlation was revealed between the age of the patient and the initial growth time and the confluence. Similarly, reverse transcription polymerase chain reaction showed that the osteoprotegerin and osteopontin expression levels did not change significantly according to the surgical technique used.In conclusion, osteotomies carried out with either instrument do not seem to substantially influence the vitality of the bone tissue. The variability of the expression levels of typical osteoblastic markers seems to be linked more to other factors than to the surgical technique used. PMID- 19816273 TI - Sudden rupture of the internal maxillary artery causing pseudoaneurysm (mandibular part) secondary to subcondylar mandible fracture. AB - The mandible is one of the most frequently fractured bones of the face. Condylar fractures are the most common mandibular fractures in adults. The potential complications of mandibular fractures are infection, paresthesia, malunion, nonunion, asymmetry, and long-term functional and growth disturbances. In this article, we report an uncommon case of life-threatening pseudoaneurysm of the internal maxillary artery secondary to subcondylar fracture. We believe that the subcondylar fracture on the vertical vector and oblique direction can damage vital tissues such as the internal maxillary artery, and this injury was, to our knowledge, the first reported acute pseudoaneurysm, which occurred just after the injury at the initial part of the maxillary artery. PMID- 19816274 TI - Staged reconstruction for adult complete Treacher Collins syndrome. AB - Treacher Collins syndrome is a congenital craniofacial malformation affecting the structures derived from the first and second brachial arches. For the complete form, the deformities can be severe; both the orbital-zygomatic region and mandible as well as the soft tissues should be reconstructed. To explore the surgical management of this kind of deformities, we proposed our protocol for the staged reconstruction: (1) upper-facial reconstruction with specially designed outer calvarial table, (2) mandibular lengthening by distraction osteogenesis technique and orthognathic surgery to correct the birdlike facial appearance and anterior open bite after distraction, and (3) lipofilling for the correction of residual depressive deformities. Two adult patients have undergone this protocol, and the results were satisfactory. Even though many authors advised using pedicled bone flap for the reconstruction of the orbit and zygomatic complex in young patient because free bone graft has the tendency of bone resorption and multiple onlay bone graft may need to be added in later stage, for adult patient, however, the reconstructed bone framework using outer cranial table can well maintain the bony facial contour of the orbital zygomatic region. The residual depressive deformities in the temporal and cheek regions can be treated with lipofilling. Compared with the technique of mandibular advancement osteotomy, bilateral mandibular lengthening by gradual distraction is relatively safe and can effectively elongate the mandible and the surrounding soft tissues. After the distraction, the open bite can be closed with subapical osteotomy, and the chin projection can be further added by advancement genioplasty. PMID- 19816275 TI - Variations of endoscopic and open repair of metopic craniosynostosis. AB - In contrast to sagittal craniosynostosis, the role of endoscopic, minimally invasive approaches in the treatment of metopic craniosynostosis with resulting trigonocephaly is not as well defined. We reviewed the senior authors' (H.M. and S.C.) clinical experience in the treatment of children with metopic craniosynostosis using a variety of endoscopic and open techniques. Thirty-three patients were treated at a single institution during a 5-year period with between 3 and 8 years of follow-up. Sixteen patients underwent 3 variations of endoscopic approaches, and 17 patients had open fronto-orbital advancement. Clinical parameters of the 2 groups were examined including age at surgery, blood loss, operative time, transfusion volume, hospital stay, complications, use of postoperative cranial banding, and the need for reoperation for persistent deformity. The various endoscopic and open techniques used by the authors in the treatment of metopic craniosynostosis are discussed in detail, including rational for individual technique selection and preliminary impressions regarding clinical outcome. PMID- 19816276 TI - Osteogenesis mechanism of chitosan-coated calcium sulfate pellets on the restoration of segmental bone defects. AB - A radial segmental defect model of a rabbit was used to study the restoration effect on defects treated with chitosan-coated pressed calcium sulfate pellets combined with recombinant human bone morphogenetic protein-2 (rhBMP-2), coated pressed calcium sulfate pellets, and uncoated pressed calcium sulfate pellets. Nothing was implanted in the control group. After 4, 8, and 12 weeks, the results indicated that coated pressed calcium sulfate pellets combined with rhBMP-2 and coated pressed calcium sulfate pellets facilitated new bone formation on defected bones and that, particularly, the coated pressed calcium sulfate pellets combined with rhBMP-2 was more effective than the coated pressed calcium sulfate pellet. Histologic and tetracycline fluorimetric findings showed that the osteogenesis mechanism of chitosan-coated pressed calcium sulfate pellets is membrane bone formation, and the pellets showed slightly slower resorption that closely coincides with the growth rate of new bone. PMID- 19816277 TI - Plating in microvascular reconstruction of the mandible: can fixation be too rigid? AB - OBJECTIVE: Determine long-term loss of mandible height with use of stress shielding reconstruction plates for free fibula flap mandible reconstruction. DESIGN: Retrospective single-blinded medical record review. SUBJECTS: Seventy patients who had fibula free flap mandible reconstructions performed for 10 years. Patients who underwent radiotherapy were excluded. METHODS: Review of 70 fibula free flap mandible reconstructions performed for the last 10 years in a city hospital revealed 7 patients (10%) who had resections for benign odontogenic diseases. All had a three-dimensional cast model made, on which the reconstruction plate was bent to the desired shape preoperatively. Free fibula height on panoramic x-ray images taken preoperatively and at 2 and 12 months postoperatively. RESULTS: Seven (10%) patients met criteria for the study. Bone height was maintained at 2 months postoperatively, but at 12 months, there was a statistically significant loss of fibular bone height averaging 20% in the anterior, body, and ramus areas (P < 0.05). Despite this, all patients were considered eligible for dental rehabilitation, and 4 of 7 patients have had osseointegrated implants placed. CONCLUSIONS: As opposed to miniplates, increased resorption may have been due to the stress-shielding phenomenon unique to a reconstruction plates. However, this did not seem to affect the ability to place osseointegrated implants. PMID- 19816278 TI - Long-term results in the bilateral cleft lip repair by Mulliken's method. AB - PURPOSE: To evaluate long-term results in the bilateral cleft lip repair by Mulliken's method, using anthropometric measurements, we assessed the growth of the nose and upper lip after the operation by comparing with those from 30 children without bilateral cleft lip. MATERIALS AND METHODS: Forty-four patients had their bilateral cleft lip and nasal deformity repaired simultaneously by Mulliken's method during the period from July 1997 to December 2007. Of these patients, 15 patients had bilateral complete cleft lip, 17 patients had bilateral incomplete cleft lip, and 12 patients had a mixed type of complete and incomplete bilateral cleft lip.To follow up on the growth of the lips and nose after the operation, the following 6 anthropometric measurements were analyzed: nasal tip protrusion, nasal width, columellar length, upper lip height, cutaneous lip height, and vermilion mucosa height. RESULTS: In most patients, nasal length, nasal tip projection, columellar length, and upper lip shape were appropriate. Nasal tip protrusion, nasal width, upper lip height, and vermilion-mucosal height were within normal limit. However, columellar length and cutaneous lip height were relatively shorter than the average values of children without bilateral cleft lip. CONCLUSIONS: By performing Mulliken's method, we can achieve natural lip and nasal shape, harmonious Cupid's bow, appropriate nasal projection, and natural philtrum. PMID- 19816279 TI - Endoscopic maxillary sinus lift without vestibular mucosal incision or bone graft. AB - AIM: To augment the thickness of the maxilla in cases with marked absorption of the alveolar bone of the upper jaw due to severe periodontal disease by performing a minimally invasive sinus lift procedure without an iliac bone graft. METHODS: An endoscope is inserted into the nasal cavity, and a large drainage opening is created in the inferior meatus extending from the maxillary sinus to the nasal cavity. The endoscope and forceps for mucosal removal are then inserted into the maxillary sinus via the drainage opening, and approximately, the lower half of the maxillary sinus mucosa is excised. RESULTS: Computed tomographic scans more than 1 year (range, 12-36 mo; mean, 23.5 mo) after performing this procedure verified maxillary sinus floor bone growth of 2.7 to 15.4 mm (mean, 7.6 mm) on 33 sides of 17 cases. Bone growth of 5 mm or more was obtained on 31 of 33 sides. CONCLUSIONS: With this procedure, it is possible to perform a sinus lift without the need for oral vestibular incision or bone graft. The results suggest it could provide a more minimally invasive approach to performing sinus lift. PMID- 19816280 TI - Mandibular deformities: single-vector distraction techniques for a multivector problem. AB - Single-vector distraction devices have been criticized for creating a malocclusion in an attempt to correct a three-dimensional mandibular deficiency, resulting in the evolution of a multiplanar device. Although there are indications for the use of a multiplanar device, a vast number of patients with mandibular hypoplasia can be effectively treated with a single-vector device, producing a normal occlusion and an aesthetic result while minimizing facial scarring and simplifying postoperative care. The purpose of this review was to describe surgical techniques whereby a single-vector device is effectively used in treating a multivector mandibular deficiency.A retrospective analysis of all patients who underwent mandibular distraction at the Children's Hospital of Philadelphia between 1996 and 2005 with a semiburied, uniplanar device was conducted. Charts, photographs, graphs, operative reports, computed tomography scans, and cephalometrograms were reviewed for those patients undergoing uniplanar mandibular distraction.Ten unilateral and 4 bilateral distractions were performed. In these 14 patients, causes included hemifacial microsomia, Treacher Collins syndrome, posttraumatic hypoplasia, and temporomandibular joint ankylosis with hypoplasia. The average device distraction was 29 mm (range, 18-34 mm). The average age at distraction was 8.4 years (range, 4-15 years). Surgical techniques for these patients will be described in detail. The single-vector, semiburied device can be effectively used to aesthetically correct a three-dimensional problem and to produce or maintain a class I occlusion while simplifying postoperative management and minimizing facial scarring. PMID- 19816281 TI - Stereolithography: a method for planning the surgical correction of the hypertelorism. AB - Hypertelorism is a deformity characterized by an increase in interorbital distance. The treatment can be orbital mobilization, or if the malformation goes along with occlusal alterations, the indicated treatment is a facial bipartition with hemifacial rotation.The intention of the present study was to describe a surgical planning technique in locating the anatomic points and planes on a stereolithographic model of a patient with 0-14 fissure and its surgical application. PMID- 19816282 TI - Nonorthodontic orthognathic surgery. AB - Orthognathic surgery for occlusal improvement commonly requires preoperative and postoperative orthodontic treatments to fine-tune intercuspation. This report focuses on the feasibility, treatment goals, and outcomes of orthognathic surgery in a series of patients with congenital dentofacial anomalies who were considered ineligible for or noncompliant with orthodontic treatment. Despite suboptimal occlusal outcomes, the orthognathic procedures resulted in tangible benefits for the patients. We conclude that surgical treatment without concomitant orthodontics is a viable option in selected patients. PMID- 19816283 TI - Double scalping flap: a versatile technique in scalp reconstruction. AB - Scalp reconstruction is a challenging area in plastic surgery. The susceptibility of potentially exposed brain tissue is almost always a concern along with the provision of adequate soft tissue coverage in cases of full-thickness defects. The applied reconstructive strategy also affects efficiency in malignant disease treatment providing both local control of the disease and vigorous monitoring for recurrence or metastasis. The general condition of the patients presenting with malignant lesions of the scalp is often impaired because of old age or concomitant disease. Therefore, demanding, long-lasting, or multistage procedures may often be undesirable. The double scalping flap comprises a 1-stage procedure, which can be used in most full-thickness defects of the vertex of the scalp. Some of the primary advantages of the double scalping flap procedure are its versatility and arc of rotation, its minimal donor site morbidity, being relatively simple, and being a short technique. Disadvantages that may be considered are the loss of hair-bearing skin at the occipital region and the poor color and texture match between scalp skin and the split-thickness skin graft that is used on the donor site. We present 2 patients with exemplary case of neglected malignancy of the scalp and their successful treatment using the double scalping flap. Comparison and contrast of alternative reconstructive procedures are also included to further investigate scalp reconstruction. PMID- 19816284 TI - Microscopic minimally invasive approach to nonsyndromic craniosynostosis. AB - PURPOSE: The purpose of this retrospective study was to present the results of the authors' microscopic minimally invasive approach in the treatment of nonsyndromic craniosynostosis. METHODS: From 2001 to 2007, the authors treated a cohort of 67 infants with nonsyndromic sagittal, unicoronal, bicoronal, and metopic craniosynostosis, either with the microscopic (n = 40) or the open (n = 27) approach. In the microscopic approach, incisions were placed over the premature suture, and using a surgical microscope, the appropriate synostectomy was performed. The open approach used a traditional coronal incision with cranial vault reconstruction. Both groups of patients had postoperative molding helmet therapy. Finally, anthropometric measurements were used to evaluate the treatment results. The measurement used for the patients with sagittal and bicoronal craniosynostoses was the divergence from the norm of the age-adjusted cephalic index. The (FZr-EUl/FZl-EUr) and (FZr-EUr)/(FZl-EUl) were used for the patients with unicoronal craniosynostosis. The divergence from the norm of age-adjusted (FTr-FTl)/(Tr-Tl) was used for the patients with metopic craniosynostosis. (FZr = right frontozygomaticus, EUl = left eurion, FZl = left frontozygomaticus, Eur = right eurion, FTr = right frontotemporale, FTl = left frontotemporale, Tr = tragion, Tl = left tragion). RESULTS: The median surgical times for microscopic and open approaches were 108 and 210 minutes, the volumes of blood loss were 75 and 220 mL, the durations of hospital stay were 2 and 4 days, the numbers of helmet were 2 and 1, and the durations of helmet therapy were 10.5 and 8 weeks, respectively. The analysis of variance for repeated measures showed that there was no statistically significant difference between the 2 groups in any of the craniosynostoses. CONCLUSIONS: The treatment outcomes from the microscopic minimally invasive approach to craniosynostosis are equal to those seen with the open approach. The microscopic approach results in less operative time, blood loss, and hospitalization. PMID- 19816285 TI - Use of bone morphogenetic proteins in sinus augmentation procedure. AB - Bone morphogenetic proteins (BMPs) have shown osteoinductive properties, and favorable regenerative responses to the graft materials containing BMPs have been reported. Maxillary sinus augmentation with placement of dental implants is a well-established technique for functional and aesthetic dental rehabilitation of partially or completely edentulous patients with severe maxillary atrophy, and attempts have been made to promote the bone formation using BMPs with different carriers in various animal models in sinus augmentation procedures.In this review, the summary was done on various carriers and the dosages for BMPs in sinus augmentation procedures. PMID- 19816286 TI - Stereological evaluation of laryngeal cancers using computed tomography via the Cavalieri method: correlation between tumor volume and number of neck lymph node metastases. AB - Tumor volume is accepted to be a significant prognostic factor for cancers arising in the head and neck regions. There were few studies on laryngeal tumor volume calculations. The purposes of this study were (1) to describe and adapt the relevant methods of computed tomography and stereology to estimate laryngeal tumor volume measured by the Cavalieri method; (2) to examine the possible correlation between tumor size and the number of involved neck lymph nodes in patients with laryngeal malignancies; and (3) to investigate if there is left right asymmetry for this correlation and the number of involved lymph nodes in bilateral laryngeal malignancies. Fifty-two patients with bilateral laryngeal cancer only from a total of 137 patients with laryngeal cancer who underwent neck dissection during the study period were included in the study. In all cases, neck dissection was part of the treatment of laryngeal cancer. The Cavalieri estimator was used to estimate the tumor volumes on computed tomographic images. The tumor volume increased with the number of involved lymph nodes in the bilateral laryngeal malignancies in both the right and left neck regions. In addition, the volume of the primary lesion and the number of lymph node metastases in the cancer cells in the right neck region were well correlated in patients with laryngeal cancer but not the number of lymph node metastases in the left neck region. Correct, unbiased, reliable volume measurements for staging and therapeutic decision making of laryngeal cancers via the Cavalieri method may be possible. PMID- 19816287 TI - Anatomic examination of the upper head of the lateral pterygoid muscle using magnetic resonance imaging and clinical data. AB - The aim of this work was to ascertain the different kinds of insertion of the upper head of the lateral pterygoid muscle (LPM) on the temporomandibular joint and to clarify its physiology to understand its possible role in the dysfunction at the temporomandibular joint. Magnetic resonance imaging examinations were used in this work to achieve a direct view of the LPM on a large number of selected patients with dysfunction at the temporomandibular joint.The study population was composed of 92 patients, of whom 74 were women and 18 were men. Their age range was from 19 to 53 years (mean age, 31 years). The images were analyzed using the following parameters: symmetry of morphologic insertions of the upper head of the LPM, types of muscular insertion divided into 3 groups (A, single bundle on the capsule and condyle; B, one bundle on the disk and a second bundle on the condyle; and C, one bundle only on the disk), and relationship between disk position regarding the condyle in patients with disk dislocations with or without reduction and upper head of the LPM. The chi-square test was used to measure the magnitude of the results.It is possible to impute to the C-type insertion morphology a negative prognostic value for a long-term improvement of disk pathology and to consider the possibility that this muscle may contribute to dislocating the disk when its insertion was directed only on the disk itself. When the upper head of the LPM was inserted on the disk, the percentage of disk dislocation without reduction was greater. PMID- 19816288 TI - Reconstruction of skull defects: currently available materials. AB - The restoration and recovery of a compromised skull continues to be a challenge to craniofacial surgeons and neurosurgeons. Different operative techniques and implant materials are being used to reconstruct the rigid framework of the skull. However, no currently available materials satisfy all of these criteria. According to this premise, the aim of this study was to report on the currently available materials for the reconstruction of the cranial vault and to describe their main characteristics, advantages, and disadvantages. Although the use of the materials discussed in this study is clearly positive for the reconstruction of skull defects and cranioplasties, there is a need for more complex studies and research into developing these materials to achieve all the ideal prerequisites stipulated by the scientific community and to evaluate their properties and aesthetic and functional results in the long term. PMID- 19816289 TI - Hyperbaric oxygen: therapy for patients with maxillofacial implants? AB - Osseointegration was an innovative treatment in dentistry during the last 3 decades. The success of osseointegration is related to factors such as material biocompatibility, adequate quality of bone tissue that allows implantation, surgical technique, and macrostructure and microstructure of implant. The osseointegrated implants are successfully applied in dental clinic including oral and facial rehabilitations mainly for patients submitted to mutilating surgeries. However, patients submitted to radiation therapy present risks to treatment with implants owing to adverse effects on bone tissue. Nowadays, the literature suggests different therapies to improve the success of osseointegration such as hyperbaric oxygen therapy that aims to prepare bone and adjacent tissues to receive the implant. Therefore, the purpose of this study was to present a literature review concerning indications, contraindications, successes, and difficulties with hyperbaric oxygen therapy associated to maxillofacial implants. PMID- 19816290 TI - Self-assembling peptide nanofiber scaffolds, platelet-rich plasma, and mesenchymal stem cells for injectable bone regeneration with tissue engineering. AB - The purpose of this study was to investigate a capability of PuraMatrix (PM), which is a self-assembling peptide nanomaterial, as a scaffold for bone regeneration in combination with dog mesenchymal stem cells (dMSCs) and/or platelet-rich plasma (PRP) using tissue engineering and regenerative technology. Initially, teeth were extracted from an adult hybrid dog's mandible region. After 4 weeks, bone defects were prepared on both sides of the mandible with a trephine bar. The following graft materials were implanted into these defects: (1) control (defect only), (2) PM, (3) PM/PRP, (4) PM/dMSCs, and (5) PM/dMSCs/PRP. From scanning electron microscope images, PM had a three-dimensional nanostructure, and dMSCs attached on the surface of PM. At 2, 4, and 8 weeks after implantation, each sample was collected from the graft area with a trephine bar and assessed by histologic and histomorphometric analyses. It was observed that the bone regenerated by PM/dMSCs/PRP was of excellent quality, and mature bone had been formed. Histometrically, at 8 weeks, newly formed bone areas comprised 12.39 +/- 1.29% (control), 25.28 +/- 3.92% (PM), 27.72 +/- 3.15% (PM/PRP), 50.07 +/- 3.97% (PM/dMSCs), and 58.43 +/- 5.06% (PM/dMSCs/PRP). The PM/dMSCs and PM/dMSCs/PRP groups showed a significant increase at all weeks compared with the control, PM, or PM/PRP (P < 0.05 at 2, 4, and 8 weeks, analysis of variance). These results showed that MSCs might keep their own potential and promote new bone regeneration in the three-dimensional structure by PM scaffolds. Taken together, it is suggested that PM might be useful as a scaffold of bone regeneration in cell therapy, and these results might lead to an effective treatment method for bone defects. PMID- 19816291 TI - Bone tissue engineering scaffolds of today and tomorrow. PMID- 19816292 TI - An enhanced strength retention poly(glycolic acid)-poly(L-lactic acid) copolymer for internal fixation: in vitro characterization of hydrolysis. AB - In general, most bioabsorbable polymers used for internal fixation are composed of copolymers of glycolic acid and lactic acid (L and/or DL). Within this family of polymers resides the latitude to design materials with specific degradation properties to meet a variety of clinical needs. One material with an extensive clinical history is LactoSorb copolymer, which is composed of a molar ratio of 82:18 poly(L-lactic acid)-poly(glycolic acid). Fixation devices derived from this copolymer have been used in orthopedic and craniofacial surgeries. With a strength loss profile of approximately 12 weeks, it mirrors the profile of normal healing; however, in cases in which delayed healing is anticipated, a copolymer with a longer strength retention profile may be desirable. This study compared the hydrolytic degradation, in vitro, of the 82:18 copolymer with a related 85:15 poly(L-lactic acid)-poly(glycolic acid) copolymer. In addition to a difference in the copolymer ratio, the latter had a larger inherent viscosity (molecular weight) and a smaller polydispersity (molecular weight distribution). This copolymer had an equivalent initial tensile modulus as the 82:18 copolymer (0.8 GPa) but a strength retention profile that spanned 44 weeks. The activation energy of hydrolysis for the 85:15 copolymer was 38.8 kcal/mol, which was greater than the 23.8 kcal/mol historical value for the 82:18 copolymer. This suggests that hydrolysis of the 85:15 copolymer is more temperature-sensitive than that of the 82:18 copolymer. These results were interpreted in the context of the differences in the compositions of these 2 copolymers. PMID- 19816293 TI - Calcium aluminate, RGD-modified calcium aluminate, and beta-tricalcium phosphate implants in a calvarial defect. AB - Calcium aluminate (CaAl), arginine-glycine-aspartic acid-modified CaAl, and beta tricalcium phosphate (TCP) implants were studied in a rat calvarial critical sized defect model. The rates of newly formed bone and osteointegration were measured using 3 different methods: radiography, micro-computed tomography, and histologic examination. After 4 weeks, there was no new bone formed and no signs of osteointegration into the skull bone in the CaAl or arginine-glycine-aspartic acid-modified CaAl groups, and thick fibrous capsules were visible around the whole circumference of the implants in both groups. In the beta-TCP group, neovascularization of the implant was observed, which is consistent with the early phase of new bone formation. In addition, in the beta-TCP group, signs of implant integration into the host tissue were evident at 4 weeks. There was no soft tissue reaction around the beta-TCP implant. These observations suggest that more specific adhesion peptides may be needed to activate the bioinert CaAl implant and promote bone formation in the craniofacial skeleton. PMID- 19816294 TI - Transforming growth factor-beta isoforms and the induction of bone formation: implications for reconstructive craniofacial surgery. AB - Craniofacial skeletal reconstruction remains a challenging problem despite major molecular and surgical developments in the understanding of bone formation by induction. The induction of bone formation has been a critical topic of research across the planet. The bone induction principle identified important cues for tissue engineering of bone, namely, osteogenic soluble molecular signals, the bone morphogenetic and osteogenic proteins, and insoluble signals or substrata including biomimetic bioactive matrices and responding stem cells. In primates, and in primates only, the osteogenic soluble molecular signals that initiate the induction of bone formation additionally include the 3 mammalian transforming growth factor-beta (TGF-beta) isoforms, members of the TGF-beta supergene family. The mammalian TGF-beta isoforms, when implanted in the rectus abdominis muscle of the nonhuman primate Papio ursinus, induce rapid and substantial endochondral bone formation resulting in large corticalized ossicles by day 30 after heterotopic implantation; in calvarial defects of the same nonhuman primates, identical or higher doses of the TGF-beta protein do not induce bone formation because of the overexpression of Smad-6 and Smad-7, gene product inhibitors of the TGF-beta signaling pathway. The addition of minced fragments of autogenous rectus abdominis muscle partially restores the osteoinductive activity of the human TGF-beta3 isoform resulting in the induction of bone formation in the treated calvarial defects. Recombinant human TGF-beta3 delivered by Matrigel matrix and implanted in class II and III furcation defects of mandibular molars of P. ursinus induce periodontal tissue regeneration. The addition of minced fragments of autogenous rectus abdominis muscle significantly enhances cementogenesis. This review highlights the induction of bone formation by the osteogenic proteins of the TGF-beta superfamily in the nonhuman primate P. ursinus and reviews combinatorial applications of myoblastic/myogenic stem cell based therapeutics for bone induction and morphogenesis. The recruitment of myoendothelial cells is also discussed in the light of the intrinsic and spontaneous induction of bone formation by smart biomaterial matrices that induce bone differentiation in heterotopic extraskeletal sites of P. ursinus without the exogenous application of the osteogenic soluble molecular signals of the TGF-beta superfamily. PMID- 19816295 TI - Effect of electromagnetic field induced by radio frequency waves at 900 to 1800 MHz on bone mineral density of iliac bone wings. AB - OBJECTIVE: Telecommunication has gained a different meaning in daily life with the introduction of the mobile phone system. However, electromagnetic pollution has increased in parallel to this improvement. In this study, we aimed to investigate the effects of electromagnetic waves emitted from cellular phones operating at a frequency of 900 to 1800 MHz on the bone mineral density of the human iliac bone wings, which are the most common carriage sites for mobile phones. MATERIALS AND METHODS: A total of 150 male volunteer participants were included in this study. The mean age was 31.85 years, and the age range was between 21 and 57 years. The participants were separated into 2 groups based on as follows: iliac side exposed to electromagnetic wave (group 1) and unexposed side (group 2). Of the total number of participants, 122 were carrying their phones on their right iliac wings, whereas 28 were carrying their phones on their left iliac wings. The mean daily carriage duration was 14.7 hours (between 12 and 20 h), and the mean duration for cellular phone use was 6.2 years (between 4 and 9 yr). Mineral bone density was measured using dual-energy x-ray absorptiometry in the right and the left iliac wings of all the participants. The SPSS 15 software (SPSS Inc, Chicago, IL) was used for statistical analysis. In the comparison of the 2 sides, Student t test was performed and P < 0.05 was considered significant. RESULTS: The mean dual-energy x-ray absorptiometry values measured from group 1 were slightly lower than those from group 2, but there was no statistically significant difference between the groups (P > 0.05). In addition, the mean values of group 1 were not as low as those measured in osteopeny or osteoporosis cases. CONCLUSIONS: Current data may suggest that taking into consideration cellular phone use when iliac bone graft is necessary in clinical practice would constitute an important factor for more favorable outcomes. PMID- 19816296 TI - Recombinant human bone morphogenetic protein-4 (BMP-4)-stimulated cell differentiation and bone formation within the expanding calvarial suture in rats. AB - Suture expansion osteogenesis, which induces active bone formation within the distracted area by mechanical stress, provides an alternative form of treatment of some bone deficiency problems in craniofacial field, such as craniosynostosis, palate cleft, or narrow maxilla. However, how to stimulate new bone formation within the distracted area remains a great challenge. In this study, the effect of recombinant human bone morphogenetic protein (rhBMP-4) on bone formation induced by mechanical stimuli was investigated. The expanded midsagittal sutures of the rat calvaria were maintained in an organ culture system in the absence (control group) or presence (experimental group) of rhBMP-4 (25 or 50 ng/mL). Proliferating cell nuclear antigen, alkaline phosphatase (ALP) activity, and messenger RNA levels of core-binding factor alpha 1 (Cbfa1) and ALP were detected to determine the cell proliferation and differentiation. Bone formation within the suture was evaluated by histologic and fluorescent examination. No difference in the number of proliferating cell nuclear antigen-positive cells was found between the control and the experimental groups, whereas ALPase activity and messenger RNA levels of Cbfa1 and ALP in the experimental group were higher than that in the control group. Bone formation was accelerated in the rhBMP-4-treated group when compared with the control group. In addition, the amounts of bone formation and cell differentiation in response to 25 ng/mL of rhBMP-4 were almost equal to those induced by 50 ng/mL of rhBMP-4. Our results suggest that application of exogenous rhBMP-4 could enhance new bone formation within the rapid expanded sutures by stimulating osteoblast differentiation. PMID- 19816297 TI - Treatment of alveolar cleft performing a pyramidal pocket and an autologous bone grafting. AB - Alveolar cleft repair is a debate topic in cleft lip and palate treatment.The aim of this article is to analyze the outcomes and the advantages of the autologous bone grafting performed during the period between 1981 and 2006. In our plastic surgery unit, 468 patients with alveolar clefts have been treated. According to our protocol, the timing for the closure of the alveolar cleft ranged from 7 to 11 years (mean, 9.4 years). Autologous bone was taken from the skull in the 45% of patients, from the iliac crest in 35% of cases, and from the chin in 20% of cases. The surgical technique of creating a pyramidal pocket to secure the bone graft was central to achieving a good result. The postoperative evaluation of the results, using clinical criteria and endoral radiography, orthopantomography, and teleradiography at 3, 6, 12 months after surgery, and more recently, in the last 82 cases by a three-dimensional computed tomography, allows us to assert that we obtained optimal results in 50% of treated cases, good results in 40%, sufficient in 4%, partial failure in 5.4%, and complete failure in 0.6%. PMID- 19816298 TI - Techniques on modeling the vascularized free fibula flap in mandibular reconstruction. AB - In mandibular reconstruction with bone grafts, the functional and aesthetic outcomes are clearly influenced by reconstructive techniques. Vascularized free fibular flaps are considered the most suitable choice for mandible reconstruction because of their good aesthetic and functional outcomes. Several techniques have been described in literature. According to various authors, shaping of the fibular graft can be performed by means of computer-aided design/computer-aided modeling procedures for the evaluation of the presurgery anatomy and three dimensional models of fibula graft. However, these models are expensive, require a strict realization of the programmed intervention, and are not easily adaptable to a surgical treatment different from the planned one. We report our experience on a technique for mandibular reconstruction using a free fibular flap, obtained by modeling a template on the resected mandible. Our operative technique consists of 5 steps: preplating, resection, template modeling, contouring of the fibula, and fixing the flap on the mandible.Template modeling is performed to record the length of the resected bone, the mandibular curvature on the 3 space planes, and the width of the basal bone. On the basis of the template, the fibula flap was modeled while still attached to the pedicle. The plates, positioned at the level of the fibular osteotomies, were preformed to make the fibula precisely fit in the template. Once the fibula was shaped, mandibular reconstruction was completed by positioning the fibula graft on the mandibular stumps. Bone graft modeling by means of template is a rapid, safe, and flexible method to reconstruct the mandible. PMID- 19816299 TI - Coronal incision with preserved temporoparietal fascia. AB - BACKGROUND: The conventional or zigzag bicoronal incision has been widely used in craniofacial surgery. Unfortunately, after the incision, the temporoparietal (TP) fascia has to be abandoned because of its vascular or structural damages. OBJECTIVE: The author describes a technique to preserve the TP fascia during the incision. METHODS: The coronal incision stopped at the subcutaneous plane. The dissection continued subcutaneously under the posterior edge of the incision as in the elevation of TP fascia. After cutting through the TP fascia, the dissection continued anteriorly in the subgaleal plane as in the coronal dissection. RESULTS: A total number of 15 craniofacial patients underwent bicoronal incision with this technique. The time spent for preservation of the TP fascia was 10 to 15 minutes for both sides. CONCLUSION: The bicoronal incision with preserved TP fascia as described provides an alternative to the patient who may need TP fascia for future uses. PMID- 19816300 TI - Important aspects of oral lining in unilateral cleft lip repair. AB - To achieve an aesthetic lip in cleft lip repair, central fullness and slight eversion of the vermilion are necessary. If only cutaneous anthropometric length is considered, symmetry and good vermilion contour may be obtained, but a seemingly tightness of the lip can occur. To prevent this, it is necessary to obtain sufficient central mucosal tissue of the oral lining.The authors used 2 methods to obtain adequate tissue of the central area of the oral lining. First, the mucosa of the central area of the oral lining was supplemented using a medial mucosal flap, and the amount of superfluous tissue was minimized. Second, a relaxing incision was placed at the oral lining of the lateral flap, which was subsequently centrally advanced.A total of 389 patients with a unilateral cleft lip underwent surgery using these methods and achieved satisfactory results. Occasional cases of lateral vermilion bulging were encountered during long-term follow-up, but these were easily corrected by bulging excision.Consideration of the oral lining is essential in cleft lip repair. The authors were able to reconstruct an aesthetically pleasing lip with central fullness by obtaining an adequate amount of tissue in the central area of oral lining. PMID- 19816301 TI - Distraction osteogenesis of the palate: an experimental model. AB - Although most cleft palates can be closed with conventional mucoperiosteal flap techniques, the occasional wide cleft or difficult fistula has few options for an early 1-stage reconstruction. Distraction osteogenesis (DO) has the potential to close the palate with both hard and soft tissues as well as mitigating the possibility of future oronasal fistula. A right unilateral 5-mm cleft was surgically created in 15 New Zealand white rabbits. In group 1 (N = 5), no further repair was performed (NR); in group 2 (N = 5), mucoperiosteal flaps were used to close the cleft for a soft-tissue-only repair (STR); in group 3 (N = 5), a unilateral osteotomy in the palate on the noncleft side allowed distraction of the palatal shelf across the cleft until closed (DO). Clinical examination, Micro computed tomography bone density, direct cephalometry, and histology were evaluated at 8 weeks after the completion of distraction. Bone mineral density (BMD; mg/mL) data were obtained from micro-computed tomography scans of both the cleft and noncleft sides for each rabbit, and a ratio was obtained [(BMDc/BMDnc) x100]; NR = 1.38, STR = 44.27, DO = 88.36, P = 0.007. Facial measurements revealed no growth disturbances as a result of DO. Histologic evaluation revealed increased organization of new bone in DO group compared with NR and STR. Clinically, DO group rabbits did not show any increase in feeding disturbances, infection, or wound healing. The success of membranous facial bone distraction has been applied to a new model for palatal repair with the potential to ameliorate the problems associated with soft-tissue-only repair. PMID- 19816302 TI - Microscopic relation of palatopharyngeus with levator veli palatini and superior constrictor. AB - The aim of this study was to elucidate microscopic relation between the levator veli palatini, palatopharyngeus (PP), and superior constrictor (SC) muscle in transverse, parasagittal, and coronal sections.In 10 Korean adult cadavers, the entire soft palate was removed, trimmed, and preserved in 10% neutral buffered formalin. Specimens were embedded in paraffin and sectioned at a thickness of 10 mum. Blocks were cut the course of levator veli palatini. Also, transverse, parasagittal, and coronal section were made, stained using Masson trichrome, and observed under light microscope.Levator veli palatini was inserted between mucous gland anteriorly and musculus uvulae posteriorly in the midline of the soft palate, where they interdigitated with those in the contralateral side. Palatopharyngeus originated from the palatine aponeurosis and posterior mucosa of the soft palate. Most of the fibers of the PP did not cross the midline in their origin; however, some fibers interdigitated across the midline. As PP went downward and crossed the levator, it divided into anterior fasciculus and posterior fasciculus. Anterior fasciculus was thick and went downward along the anterolateral side of the levator. Posterior fasciculus was thin and widely spread along the posteromedial side of levator. Below the levator, 2 fasciculi united and were inserted to the medial side of SC. There were close attachment between the PP and SC.A detailed understanding of the microscopic relationship between the levator veli palatini, PP, and SC muscle is desirable for performing pharyngeal flap surgeries. PMID- 19816303 TI - Endoscopic endonasal approach to the orbital apex and medial orbital wall: anatomic study and clinical applications. AB - OBJECTIVE: The objective of this study was to recognize the endoscopic anatomy of the orbital apex and medial orbital wall to understand the pure endoscopic endonasal approaches to this region and their clinical applications. These basic information will facilitate our surgical procedures and decrease the rate of surgical complications. MATERIAL AND METHODS: Five fresh adult cadavers were studied bilaterally (N = 10). We used Karl Storz 0- and 30-degree 4-mm, 18-cm, and 30-cm rod-lens rigid endoscopes in our dissections. After cadaver specimen preparation, we approached each orbital apex and medial orbital wall through each nostril. After resection of medial orbital wall, an endoscopic intraorbital approach was performed. RESULTS: The orbita could be exposed by using 0- and 30 degree endoscopes. We preferred to start the approach from the sphenoid sinus instead of transethmoidal approaches that are less familiar to the neurosurgeons. The posterior and anterior ethmoidal arteries are in close relation to the supralateral wall of ethmoid sinus, thus care must be taken not to injure these arteries during dissection. In this way, we can safely expose the whole medial wall of the orbita. Optic canal decompression can be safely done by bone resection starting from the optic nerve toward the optic canal. We continued bone resection from the posterior to the anterior of the medial orbital wall, thus we can perform medial orbitotomy. The intraorbital approach can be done medially by introducing the endoscope between the medial and inferior rectus muscles. CONCLUSIONS: Our anatomic study offered the facility to learn the endoscopic anatomy of the orbital apex and the medial wall of the orbita and understand the appropriate approaches (such as medial orbitotomy and optic canal decompression) to some pathologic lesions of this region. With skilled and experienced hands, it can superimpose many traditional orbital approaches with minimal invasiveness and less postoperative complications. PMID- 19816304 TI - Surgical treatment of a giant lipoma in the parotid gland. AB - Recently, a case of deep-lobe lipoma with enucleation was reported, but frozen section biopsy for the confirmation of the malignancy was not done. It has been suggested that lipoma in the deeper tissues should be regarded as a well differentiated liposarcoma and be treated with wide excision. Our experience is that of a 75-year-old woman who had a mass with fat density in the deep lobe of the right parotid gland, which extended through the parapharyngeal and the buccal spaces. Lumpectomy with frozen-section biopsy was performed, not only preserving branches of facial nerve but also ruling out the malignancy. Frozen-section biopsy showed a lipomatous lesion without malignancy, so further treatment such as total parotidectomy was not needed. PMID- 19816305 TI - Paradoxical vocal cord motion--haloperidol usage in acute attack treatment. AB - Paradoxical vocal cord motion (PVCM) is an uncommon disease characterized by vocal cord adduction during inspiration and/or expiration. It can create shortness of breath, wheezing, respiratory stridor, or breathy dysphonia. Possible etiological factors include asthma, underlying psychologic condition, gastroesophageal acid reflux disease, respiratory irritants exposure, central neurologic diseases, viral upper airway infections, and postsurgical procedures. Many treatment modalities were performed for acute attack of PVCM, including reassurance and onsite maneuvers, benzodiazepines, heliox, and so forth. We report a patient with PVCM who had stridor and dyspnea for 10 days and responded to intravenous haloperidol treatment. PMID- 19816306 TI - Surgical treatment of an extraparotid pleomorphic adenoma of minor salivary glands of the cheek. AB - Only few cases of extraparotid localizations of pleomorphic adenoma (PA) were reported in literature: at the lips, tonsils of the tongue, the occipital area and of the nasal septum, retropharyngeal area, temporal bone, and the thyroglossal and pituitary ducts.The finding of a PA in the minor salivary glands of the cheek was described only in 2 cases during the last 24 years and represented an exceptional finding.The aim of this article was to illustrate a case of extraparotid PA developing at the salivary glands of cheek, a short distance from Stensen duct, and to report surgical strategy. PMID- 19816307 TI - Isolated congenital palatal fistula without submucous cleft palate. AB - Congenital fistula of the palate is a rare deformity. It has been generally associated with cleft palate. Treatment of cleft palate is surgical intervention. We present a child with congenital fistula of palate that was not associated with submucous cleft and closed spontaneously at 18 months. PMID- 19816308 TI - Gossypiboma after mandibular contouring surgery. AB - BACKGROUND: Gossypiboma is derived from the Latin word gossypium, meaning cotton, and it means a postoperatively retained foreign body used in operations. Several cases of gossypiboma have been reported especially after abdominal surgery, but there has not been any reported case in plastic surgery. Mandibular contouring surgery cannot ensure a view wide enough to avoid injury to surrounding structures such as a facial artery and a retromandibular vein. In addition, many surgeons pack the sponge into the operative field to prevent bleeding, and surgeons may neglect remnant surgical materials. Recognition of gossypiboma is essential but is often considerably delayed and cause medicolegal problems. Therefore, it is important to ensure that every effort is made to prevent such occurrences. We had a chance to evaluate and treat gossypiboma, and in this paper, we want to share our experiences. MATERIALS AND METHODS: In circa 1999 to 2007, there were 3 cases diagnosed as gossypiboma after a mandible angle surgery. All patients were female, and some had signs of fever, swelling, tenderness, and purulent discharge of an oral wound. We performed a computed tomographic scan and blood test, and foreign body removal was done under general anesthesia. Intraoperatively, the diagnosis of gossypiboma was confirmed. RESULTS: All symptoms were reduced or subsided after surgery. It was noted that no postoperative infection remained. CONCLUSIONS: Gossypiboma must be considered when fever, unilateral swelling, tenderness, or unhealed oral wound is sustained despite an antibiotics therapy and a drainage procedure after a mandible angle surgery. In that case, a computed tomographic scan can be recommended as an effective method for detection of gossypiboma. PMID- 19816309 TI - Bleeding from posterior superior alveolar artery in Le Fort I fracture. AB - This article describes a patient in whom significant oronasal bleeding developed after an injury to posterior superior alveolar artery of the maxillary artery in Le Fort I fracture.A 34-year-old man had a facial injury after an explosion of a furnace. Computed tomography demonstrated Le Fort I fracture, right open zygomatic tripod fracture, and open nasal bone fracture. Blood pressure, hemoglobin, and hematocrit levels had fallen to 110/60 mm Hg, 5.7 mg/dL, and 16.1%, respectively, 10 hours after injury despite continuous blood transfusion. Selective digital angiography confirmed an injury to the posterior superior alveolar artery, with extravasated pooling of contrast material in the maxillary sinus. The distal internal maxillary artery was embolized with n-butyl cyanoacrylate and lipiodol. Postembolization angiogram showed resolution of the contrast leak, and the patient's oronasal bleeding resolved.We suggest that if the oronasal bleeding continues in Le Fort fracture, bleeding from the posterior superior alveolar artery should be suspected. In case the vital sign is not stable, selective angiography should be performed before surgery. PMID- 19816310 TI - Isolated lower lip fistulas in Van der Woude syndrome. AB - Van der Woude syndrome (VWS) is a dominantly inherited disease of orofacial region. Characteristic features of this syndrome are bilateral lower lip sinuses along with cleft lip or palate deformity. However, isolated lower lip pits in VWS without any cleft syndrome is uncommon. Lip pits in VWS are usually asymptomatic; however, patients may complain of watery drainage and/or infection. In this report, asymptomatic isolated lower lip sinuses without any cleft syndrome in a patient and his father are presented. PMID- 19816311 TI - Prosthetic rehabilitation of a patient with a mandibular defect caused by a gunshot wound. AB - Injuries to the orofacial region may vary from localized injuries to extensive soft and hard tissue loss. In addition to physical and psychologic damages, functional and aesthetic aspects must be restored. This clinical report describes the rehabilitation of a patient with a mandibular defect caused by a gunshot wound. Rehabilitation of this patient, with the use of an overdenture prosthesis, was performed after mandibular surgical hard and soft tissue reconstruction. A maxillary total prosthesis and an implant-supported mandibular overdenture supported by 4 osseointegrated implants were fabricated. Despite limited mouth opening and anatomic deficiencies, the patient's aesthetic and functional demands were fulfilled. PMID- 19816315 TI - Rhinorrhea as a rare but disturbing complication of rhinoplasty. PMID- 19816316 TI - Avoiding unanticipated adverse effects of recombinant human bone morphogenetic protein-2 therapy in craniofacial surgery with experiences from spinal applications. PMID- 19816317 TI - Effects of preoperative local ropivacaine infiltration on postoperative pain scores in infants and small children undergoing elective cleft palate repair: statistical considerations to transform interesting hypotheses into significant conclusions. PMID- 19816318 TI - The crafts of cleft surgery, advances made, and progress felt. PMID- 19816319 TI - A passion for excellence. PMID- 19816320 TI - ICPF cleft 2008 Dallas, TX: alligator swamp. PMID- 19816321 TI - Introduction of Prof. Tisheng Chang: the pioneer of plastic and craniofacial surgery in China. PMID- 19816322 TI - Current status of cleft lip and palate management in China. AB - Several million individuals with cleft lip and palate (CLP) exist in China, and it is estimated that at least 30,000 newborns with CLP are born every year. How to give them an optimal treatment with excellence is still a problem in most areas. The treatment of cleft patients presents various situations around world that are different from those in the United States and Europe. In this article, we reviewed the current status of CLP management in our country and shared our experience from one of the biggest CLP centers in China. We hope the management of cleft can be improved, and more patients can be treated by dedicated teams and dedicated centers in China. PMID- 19816323 TI - My views on cleft lip and palate care in China. PMID- 19816324 TI - Long-term solutions for the delivery of craniofacial care in India and the developing world. AB - An international group of craniofacial surgeons was consulted to identify means of providing care of consistent quality to children with craniofacial anomalies in India and subsequently use this as a model for delivering craniofacial care to children in need throughout the developing world. Three senior craniofacial surgeons were selected from the Medical Advisory Board of the World Craniofacial Foundation to evaluate eligible centers in India. A subcommittee of 10 surgeons from the World Craniofacial Foundation Medical Advisory Board was formed to help develop summary recommendations based on this site visit. These centers were selected after contacting plastic surgeons and orthodontists involved in craniofacial and cleft care throughout India and asking the director of each center to complete a questionnaire documenting their current level of activity. Based on these responses, 3 potential craniofacial centers in India were identified. These sites were selected as potential locations for the development of craniofacial centers that could provide free care to indigent patients who otherwise could not receive treatment. The sites chosen consisted of a private hospital (southern India), a state-run government hospital (central India), and a federal government hospital (northern India). After the site visits, the directors of each of the 3 potential craniofacial centers in India were asked to complete a questionnaire helping to further identify their specific needs. The subcommittee of the World Craniofacial Foundation Medical Advisory Board then developed a set of summary recommendations to help guide the development of regional craniofacial centers in India. We feel that this process will facilitate access to care to indigent children with craniofacial anomalies who would not otherwise be eligible to receive appropriate care. PMID- 19816325 TI - Why do we behave as we do? AB - It is important for surgeons to have insight into themselves, their life stories, and the rationales they use to convince themselves that their actions are unselfish and well motivated. The battle between Philanthropia and Philotechnica was recognized by Hippocrates and is still a source of internal strife for many surgeons: the need to perform an operation that they are poorly equipped to do offset against the knowledge that it could be better done by someone else. In the treatment or referral for treatment of children with craniosynostosis and craniofacial syndromes, appropriate referrals are often not made or are made only after some problem has occurred as a result of surgical intervention. Several instances of children receiving extensive surgery for wrongly diagnosed craniosynostosis are explored. The thesis is that only by knowing our own internal motivations can we avoid falling into a posture that is good for our own egos and pocketbooks but bad for our patients. PMID- 19816326 TI - Genetics and human malformations. AB - Genetics gains more and more importance in all areas of health care including craniofacial surgery and dentistry. This does not mean that every patient will benefit from genetic advances, but for many health problems, we will see progress in explaining disease pathogenesis, establishing diagnosis, guiding therapy, predicting prognosis, and achieving prevention. In this report, we briefly review the roles of the PAX9, MSX1, AXIN2, and EDA genes in the causation of congenital tooth agenesis and the promise of molecular genetic research for the improvement of patient care. PMID- 19816327 TI - Establishing a craniofacial center in a developing country. AB - The establishment of a craniofacial in a developing country is often difficult because of the availability of trained personnel. This requires gradually replacing or training personnel such as speech pathologists. Gradual improvement is possible for a period of 5 to 10 years to train individuals. This requires a dedicated person to achieve the goal of a multidisciplined center for patients with cleft. PMID- 19816328 TI - Strategies for achieving long-term effective outcome in cleft missions: the Noordhoff Craniofacial Foundation and Chang Gung Memorial Hospital. AB - The Noordhoff Craniofacial Foundation and Chang Gung Memorial Hospital have partnered to deliver cleft programs to developing nations still in need of adequate cleft care. The lessons learnt through the development of the Chang Gung Cleft and Craniofacial Center from humble beginnings to its current international standing have enabled insights into devising key strategies for achieving long lasting and compounding outcome in cleft missions. Close collaboration with local governing and health authorities, as well as establishment of ongoing support from charitable organizations that share similar philosophies, is an essential component to creating growth and sustainability of a cleft program. Identification of local "seed" physicians and key personnel, and their subsequent training at a major cleft center, is pivotal to the establishment of local cleft centers and cleft foundations that would ultimately empower local health care providers' autonomy in delivering the highest standard of care to patients with cleft in their own country. PMID- 19816329 TI - A model for the development and delivery of craniofacial health care systems in developing countries. AB - A model for developing and delivering health care systems for treating individuals with craniofacial deformities in developing countries requires defining processes that are manageable within the contexts of governments, hospitals, health care environments, and local communities.The model presented in this article is based on 6 principles that have been defined and refined by working in 10 countries since 1975. The 6 principles are designed to enhance quality of life and issues of equity within society for patients with facial deformity. The model also promotes the importance of professional equity within multidisciplinary health care teams. PMID- 19816330 TI - Developing and standardizing a center to treat cleft and craniofacial anomalies in a developing country like India. AB - The range of facial deformities is enormous. All produce some degree of disfigurement and result in the impairment of function to some degree, sometimes even to the point of incompatibility with life. Congenital facial defects in India are associated with considerable superstition, social rejection, and failure to integrate into society.In India, cleft defects occur in 1 in 500 births. Congenital facial defects are a pressing problem in India owing to the limited resources to treat such patients. Poverty is a major factor for parents of such children to get appropriate treatment.Setting up an institute to treat children with cleft and craniofacial deformities in India presents problems with financing treatment for poor patients, procuring the right infrastructure, and employing well-trained human resources.The authors have set up such an institute in Hyderabad in the southern state of Andhra Pradesh in India. The logistics of setting up such a facility in a developing country and the future of funding for cleft treatment are important factors to consider while establishing a center for patients with cleft and craniofacial anomalies.The aim of setting up such centers was to provide quality comprehensive treatment for patients from all sections of society with cleft and craniofacial anomalies. PMID- 19816331 TI - Team care for the patient with cleft: UCSF protocols and outcomes. AB - To meet the medical, dental, and psychologic needs of the individual with a complex craniofacial birth defect such as cleft lip and palate, a team of specialists must be involved. The currently accepted standards of care can only be met if the appropriate specialists work together in the diagnostic workup and in the immediate and long-term planning and execution of the various treatment modalities required. This concept is clearly stated in the "Parameters" document published by the American Cleft Palate-Craniofacial Association in 2004 (Cleft Palate Craniofac J 1993;30[Suppl]:S1-16).Not only does the interdisciplinary team care provide the best overall outcomes for the patient but also is the most efficacious and cost-effective way of meeting the goals of treatment. Interdisciplinary cleft team members should also be a resource in educating parents, students, and other health care providers and may be participants in research on prevention, interception, or regeneration. We present in this article the protocols and treatment outcomes of the University of California at San Francisco. PMID- 19816332 TI - A longitudinal cephalometric study from age 5 to 18 years on individuals with complete bilateral cleft lip and palate. AB - Development of the facial profile between the age of 5 years and the end of pubertal growth in patients with complete bilateral cleft lip and palate was studied by means of cephalometric radiographs obtained at the age of 5, 10, and 15 years as well as at the end of the growth period. All 29 patients had been treated according to the same plan and operated on by the same surgeon.Values for the ANB angle are typically very high in 5-year-old children and then decrease to values corresponding to those of the general population by the end of the pubertal growth period. Vertical development maintains the initial pattern; no compensatory vertical excess was observed. Owing to the wide range in all measurements, age-related mean values in a patient sample are of little consequence for individuals but can point out tendencies.The multidisciplinary concept of maintaining the initially protrusive position of the premaxilla by means of a passive plate at the newborn and infant stage, as well as using surgical procedures with limited retrusive effect, proved to be correct in the long run: At the young adult stage, the ANB angle remained positive for almost all patients except for those with multiple-tooth agenesis in the upper arch. A potential benefit of 2-stage palate repair is seen in the neutral vertical development of the patients examined. This hypothesis is supported by comparing with other studies from the literature. PMID- 19816333 TI - Reference photographs for nasolabial appearance rating in unilateral cleft lip and palate. AB - A popular method for nasolabial rating in unilateral cleft lip and palate (UCLP) is the Asher-McDade system consisting of a 5-point ordinal scale assessing nasal form, nasal symmetry, nasal profile, and vermilion border. The aim of the current study was to identify reference photographs illustrating this scale to facilitate its use.Four observers assessed nasolabial appearance on frontal and profile photographs of the nasolabial area of 42 children of Caucasian origin with a repaired UCLP at age 9 years. Cronbachs alpha, based on the individual scores of the 4 observers, ranged from 0.73 to 0.82 for the 4 nasolabial ratings, indicating a good reliability. The reliability of the overall score (mean of the 4 component scores) was also high (Cronbachs alpha, 0.83). Both for the nasolabial component ratings and for the overall score, duplicate measurement errors were small. The reliability for the mean of the 4 observers' scores was good, Spearman rank correlation coefficients ranging from 0.56 to 0.96.Subsequently, photographs were selected that showed the highest agreement among observers. For each of the 4 components (eg, nasal form, nasal deviation, nasal profile, and shape of the vermilion border), 5 photographs were selected to illustrate the whole range of the scale (score, 1-5), resulting in the selection of 20 pictures.It was concluded that nasolabial appearance rating can be performed reliably using a panel of judges and averaging the scores of all observers. Reference photographs, as developed from this study, may facilitate the rating task. PMID- 19816334 TI - Interdisciplinary evaluation of consecutive patients with unilateral cleft lip and palate at age 6, 15, and 25 years: a concurrent standardized procedure and documentation by plastic surgeon; speech and language pathologist; ear, nose, and throat specialist; and orthodontist. AB - PURPOSE: To evaluate surgical results, speech, hearing, and craniofacial morphology after primary cleft repair performed from 1973 to 1979. METHODS: During the years 1972 to 1985, all primary cleft surgeries were performed by 1 plastic surgeon, using Tennison lip closure combined with a periosteoplasty on the clefted alveolus at age 3 months. By mobilizing mucoperiosteal flaps, bony bridges were induced in the alveolar process in approximately 60% of the cases. All patients had the soft palate closed at age 24 months by a pushback technique. All children with complete unilateral clefts without soft tissue bands (unilateral cleft lip and palate) primary operated on 1973 to 1979 were included in the material, except 3 patients with a syndrome and 2 patients of foreign ethnicity. The material involved 30 consecutive patients with unilateral cleft lip and palate (20 boys and 10 girls; 16 left-sided and 14 right-sided clefts) who were operated on. Standardized records including photos, radiographs, lateral cephalograms, plaster model, and recording of speech and hearing were collected according to the treatment protocol at age 6, 15, and approximately 25 years. RESULTS: Evaluation included craniofacial descriptive cephalometric analysis, dentoalveolar morphology, dentofacial aesthetics, speech concerning articulation and nasality, and hearing status. Number of surgical interventions after primary surgery was recorded. Secondary revisions and candidates for orthognathic surgery are reported. Ten consecutive patients (case nos. 11-20) are demonstrated as clinical reports. CONCLUSIONS: : This longitudinal study shows how a multidisciplinary evaluation adequate for intercenter comparison can be performed when standardized procedures, registrations, and documentations are available. PMID- 19816335 TI - Characterization of the maxillary complex in unilateral cleft lip and palate using cone-beam computed tomography: a preliminary study. AB - Deficiencies in maxillary growth have often been implicated in suboptimal results of cleft lip and palate therapy. Cone-beam computed tomography provides an opportunity to look at the dimensions of the maxillary complex in three dimensions in a way that is not possible with lateral cephalograms or dental models. The purposes of this preliminary study were to outline a new set of 18 cone-beam computed tomographic measurements, apply them to 6 patients with unilateral cleft lip and palate (UCLP), and contrast them to a comparable sample of 7 normal young adults. The patients with UCLP were treated with a single protocol by a single surgeon and orthodontist. The 18 measurements had a mean intrarater reliability of 0.95 and ranged from 0.40 to 2.23 for the individual measurements. The mean interrater reliability was 1.01 and ranged from 0.40 to 2.45 for the individual measurements. Significant differences between the patients with UCLP and control subjects (combined sex samples) were found in palate length, anterior palate thickness, overall sagittal maxillary length, and premaxillary height (Mann-Whitney U tests, P